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This section contains a directory of research groups in the Animal Health field with information on their research lines and projects, services, facilities, patents, publications, the contact of the main researcher, etc.

Directorio de grupos de investigación en Sanidad Animal

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How to improve global management of Crimean-Congo hemorrhagic fever by 2030?

A group of global experts of which José de la Fuente (SaBio, IREC) is a member updates the World Health Organization`s research and development roadmap for Crimean-Congo hemorrhagic fever, and notes that effective management of the disease by 2030 will only be achieved through coordinated international action by affected countries based on a “One Health” approach.

Livestock and One Health: technological innovation for open air farms

A One Health study demonstrates that non-invasive sampling facilitates biosecurity surveillance in outdoor livestock farms and suggests that the richness of vertebrate wildlife may limit the circulation of multi-host pathogens.

Levels of blood lead in Griffon vultures from a Wildlife Rehabilitation Center in Spain (González et al., 2017).

Lead is a highly toxic contaminant that significantly impacts bird wildlife, with Griffon vultures (Gyps fulvus) serving as sensitive indicators of environmental contamination due to their position at the top of the food chain and dependence on human-related activities. This study aimed to assess lead intoxication susceptibility in Griffon vultures admitted to Wildlife Rehabilitation Centers (WRC) by measuring blood lead levels and examining correlations with clinical signs, hematological, biochemical, or radiographic findings. Factors such as age, gender, body condition, season, and primary cause of admission were also considered. Blood lead levels were measured using anodic stripping voltammetry in vultures admitted over one year to the GREFA Rehabilitation Center. Results showed that 26% of the birds had lead levels above 20 µg/dL, while 74% had levels below 20 µg/dL (mean [Pb]<20 = 9.34±5.60 µg/dL). Statistically significant differences in lead levels were found based on sex, season of admission, and body condition. A negative correlation was observed between lead levels and hematocrit. However, no direct association was found between clinical signs and blood lead levels, except for digestive issues like stasis and weight loss. While lead poisoning in vultures is often linked to ingestion of lead ammunition, no radiographic evidence of lead was found in this study. Compared to other studies, the blood lead levels detected in this population were relatively low, though all birds had detectable lead concentrations. The vultures appeared to have a higher sensitivity to the toxic effects of lead than previously reported, and there is evidence that subclinical lead exposure may predispose them to injury or disease, even if admitted for other reasons.

Monitoring protected areas from space: A multi-temporal assessment using raptors as biodiversity surrogates (Regos et al., 2017)

Monitoring protected areas (PAs) is vital for assessing their effectiveness in habitat protection, preservation, and representativeness. This study explores how species distribution models, integrating remote sensing data and biodiversity surrogates, can help create a systematic monitoring protocol for PAs. Specifically, the study evaluated the effectiveness of the Natura 2000 (N2000) network in conserving seven raptor species in a dynamic landscape in northwest Spain from 2001 to 2014. It also analyzed the cost-effectiveness of the N2000 network, using the total protected area as an indicator of conservation costs. The findings revealed that N2000 poorly represented raptor habitats. However, despite the low representativeness, the network was highly effective in increasing habitat availability for both generalist and forest specialist species during the study period. To improve representativeness, additional protected areas should be designated in the future to safeguard open-habitat specialists and their priority habitats. Proactive conservation efforts in natural and semi-natural ecosystems, such as montane heathlands, will be crucial for the long-term protection of Montagu’s harrier (listed in Annex I of the Bird Directive) and to meet EU environmental regulations. The study highlights how the integration of freely-available satellite data and species distribution models can enhance the cost-efficiency of PA monitoring systems and support the ´Fitness Check´ process of the EU Nature Directives.

Immature and mature sperm morphometry in fresh and frozen-thawed falcon ejaculates (Villaverde-Morcillo et al., 2017)

Sperm morphometry can help predict fertility and sperm freezability in various species. Understanding the ejaculate`s sperm characteristics and morphometric descriptors is crucial for effective sperm cryopreservation. This study aimed to describe the sperm of two falcon species (Peregrine Falcon Falco peregrinus peregrinus/brookei and Gyrfalcon Falco rusticolus), including immature sperm, sperm head morphometry, and the presence of subpopulations of mature sperm. Semen samples were collected via massage and voluntary false copulation, diluted with Lake and Ravie medium, and analyzed through morphological and computerized morphometric techniques, measuring sperm head length, width, area, and perimeter. In the Gyrfalcon, pooled semen was frozen using DMA as a cryoprotectant, and analyses were repeated after thawing. The study found similarly high percentages of immature sperm in all species/subspecies: 55.5% in Brookei Peregrine Falcons, 65.5% in European Peregrine Falcons, and 64.7% in Gyrfalcons. Four distinct subpopulations of mature spermatozoa were identified, with different morphometric characteristics, although their relative proportions were consistent across species. The thawed Gyrfalcon samples showed significantly lower morphometric values compared to fresh samples. These findings support the notion of pleiomorphy in raptor sperm and suggest that identifying different sperm subpopulations may play a critical role in developing future cryopreservation protocols for falcons.

CHARACTERIZATION OF NATURAL EJACULATES AND SPERM CRYOPRESERVATION IN A GOLDEN EAGLE ( AQUILA CHRYSAETUS ) (Villaverde-Morcillo et al., 2015)

This paper describes the sperm characteristics and response to cooling and freezing of naturally ejaculated semen from a captive, adult golden eagle (Aquila chrysaetus) trained to allow sperm recovery via cooperative copulation. A basic spermiogram was prepared, and sperm motility and morphometric variables recorded using a computer-aided system. For sperm storage, the effects of a polyvinylpyrrolidone-based extender were evaluated at 5°C. The same extender was also used in freezing procedures in which glycerol (11%) and dimethylacetamide (6%) were compared as cryoprotectants. The extender preserved sperm viability over storage periods of up to 6 days. Although sperm motility and percentage live sperm values were poorer for frozen–thawed (5.8–14.6% and 44–42%, respectively) than for fresh samples (46.7 and 74.6%, respectively), no differences were seen between the effects of the two cryoprotectants. These results could be of use when attempting to store the sperm of golden eagles and other raptors.

Trichomonas gypaetinii n. Sp., a new trichomonad from the upper gastrointestinal tract of scavenging birds of prey (Martínez-Díaz et al., 2015).

In the context of an epidemiological study carried out by several wildlife recovery centers in Spain, trichomonads resembling Trichomonas gallinae were found in the oropharyngeal cavity of 2 Egyptian vultures (Neophron percnopterus) and 14 cinereous vultures (Aegypius monachus) which did not show any symptoms of trichomonosis. In order to characterize them, these isolates along with seven other T. gallinae isolates obtained from different hosts and from different geographical origin were analyzed. Genetic analyses were performed by sequencing the small subunit ribosomal RNA (SSU-rRNA) and the internal transcribed spacers (ITS1 and ITS2) and the 5.8S rRNA regions. The morphological study of the isolates in both light and scanning electron microscopy was also performed. The sequences obtained in the genetic analysis coincide with previously published sequences of an isolate named as Trichomonas sp., obtained from a bearded vulture (Gypaetus barbatus), and showed clear differences to the T. gallinae sequences (97 and 90–91 % homology, respectively, for SSU-rRNA and ITS regions) and display higher similarity with Trichomonas vaginalis and Trichomonas stableri than with T. gallinae. Multivariate statistical analysis of the morphometric study also reveals significant differences between the trichomonads of vultures and the isolates of T. gallinae. The isolates from vultures presented smaller values for each variable except for the length of axostyle projection, which was higher. These results together with the different nature of their hosts suggest the possibility of a new species of trichomonad which we hereby name Trichomonas gypaetinii, whose main host are birds of the subfamily Gypaetinae.

Detection and assessment of electrocution in endangered raptors by infrared thermography (Melero et al., 2013)

Most European birds of prey find themselves in a poor state of conservation, with electrocution as one of the most frequent causes of unnatural death. Since early detection of electrocution is difficult, treatment is usually implemented late, which reduces its effectiveness. By considering that electrocution reduces tissue temperature, it may be detectable by thermography, which would allow a more rapid identification. Three individuals from three endangered raptor species [Spanish imperial eagle (Aquila adalberti), Lammergeier (Gypaetus barbatus) and Osprey (Pandion haliaetus)] were studied thermographically from the time they were admitted to a rehabilitation centre to the time their clinical cases were resolved.

Avian predators as a biological control system of common vole (Microtus arvalis) populations in north‐western Spain: Experimental set‐up and preliminary results (Paz et al., 2013).

Ecologically based rodent pest management using biological control has never been evaluated for vole plagues in Europe, although it has been successfully tested in other systems. The authors report on the first large-scale replicated experiment to study the usefulness of nest-box installation for increasing the breeding density of common kestrels (Falco tinnunculus) and barn owls (Tyto alba) as a potential biological control of common vole (Microtus arvalis) abundance in agricultural habitats in north-western Spain. The results show that: (1) population density of both predator species increased in response to both nest-site availability and vole density; (2) voles are a major prey for the common kestrels during the breeding period; (3) vole density during the increase phase of a population cycle may be reduced in crop fields near nest boxes. The installation of nest boxes provides nesting sites for barn owls and kestrels. Kestrel populations increased faster than in areas without artificial nests, and the common vole was one of their main prey during the breeding season. The results suggest that local (field) effects could be found in terms of reduced vole density. If so, this could be an environmentally friendly and cheap vole control technique to be considered on a larger scale

Occurrence of Chlamydiaceae in non-symptomatic free-living raptors in Spain (Ortega et al., 2012)

Few studies have investigated the role of raptors as natural reservoirs of Chlamydiaceae spp. and the preferred anatomical sites where these bacteria can be detected in non-symptomatic wild birds. We investigated the occurrence of Chlamydiaceae in 54 non-symptomatic adult free-living birds belonging to 14 species sampled upon reception in a raptor rehabilitation centre in Spain, and ten juvenile birds from five species born and reared in the centre for subsequent release into the wild. Swabs from conjunctivae, choanae and cloacae were taken to detect Chlamydiaceae DNA by a family-specific polymerase chain reaction (PCR) and a nested genus- and species-specific PCR. Chlamydiaceae DNA was detected in adult raptors belonging to 12 species (85.7%), mainly in conjunctival (40.6%) and, to a lesser extent, choanal (17.2%) swabs, but never in cloacal samples. Neither the genus nor the species of Chlamydiaceae could be confirmed by the nested PCR assay. Our results suggest that most of the raptor species investigated, especially the Common Kestrel (Falco tinnunculus) and Eurasian Griffon Vulture (Gyps fulvus), can be natural reservoirs of Chlamydiaceae spp. in the wild. Failure to identify the organisms at genus and species level might have been due to the poor quality and low concentration of DNA in the samples or to the presence of hitherto unclassified Chlamydiaceae species.

Life SOS Pigargus

Life Rhodope Vulture

Life Abilas

WILDBIOPHAGE

Proyecto Gypaetus Monachus

Aquila-a-Life

BIOPROTECT

Biological control of common vole pests

Life Bonelli

Environmental Training for Workers in the Hunting Sector

Using natural predators for biologic control of common vole

Vulturnet

Population management and medicine

Health surveillance of wild species in the environment by collecting samples for analysis in a “One Health” approach. Conservation Medicine: reinforcement, reintroduction and conservation programs for protected species.

Parasitology

Parasitological diagnosis of animals admitted to GREFA’s wildlife center. Molecular analysis and characterization of pathogens of importance for wildlife and public health such as Trichomonas spp. or Chlamydia spp. Diagnosis and characterization of haemoparasites in animals admitted to the center.

Post-mortem studies

Post-mortem diagnosis of cause of death in wild species. Development and improvement of necropsy protocols, carcass utilization and external anatomopathological studies. Diagnosis of environmental crimes against wildlife.

Toxicology

Analysis of the presence of microplastics in wild species.

Clinical microbiology

Clinical diagnosis of animals admitted to GREFA’s wildlife center. Study of bacteria of public health importance such as Salmonella spp., Campylobacter spp. or Escherichia coli. Detection of antibioresistance in wildlife.

Blood analysis

Clinical diagnosis of animals admitted to GREFA’s wildlife center (hematology, biochemistry and proteinogram), as well as animals that are part of in situ projects. Determination of reference values and study of the health status of wild animal populations.

The paradox of the commercial hunting of red-legged partridges

Study suggests that game lands devoted to the commercial hunting of red-legged partridges can act as “conservation islands” with positive effects on biodiversity beyond their borders

Combination of water activity and pH to improve food safety in microbiological quality outcomes.

Study of livestock feeding for the improvement of food safety in microbiological quality results

Temporal evolution of antibiotic levels in pork meat, viscera, and fatty tissues to ensure compliance with legal requirements of countries outside the European Union

Search for natural alternatives to preservatives and antioxidants that are precursors of nitrosamines

Factors influencing the curing conditions of beef and new optimization alternatives for the process.

Several quality parameters of beef have been determined over a 31-day maturation period, applying two different methods: dry maturation and wet maturation. The variation in humidity, pH, tenderness, and volatile profile has been analyzed every 2-3 days throughout the maturation period. The loss of moisture should be regarded as a decrease in the weight and size of the product, which impacts the producer`s interests. Based on the obtained results, a principal component analysis has been conducted to determine the optimal maturation moment, where tenderness and desirable aromas are maximized, while undesirable aromas of raw meat are minimized.

Optimization of quality and food safety parameters associated with the improvement of welfare in slaughter animals in an organic farming system.

The objective of this project is to investigate and compare the amounts of flavor precursor compounds present in pork samples from three different types of pigs (white breed pigs from organic and conventional farming, and Duroc pigs), taking triplicate samples from three distinct parts of the animal (leg, loin, and tenderloin). The purpose of this study is not to identify which samples have a more pleasant or unpleasant flavor, as the most reliable way to conduct such assessments today is through expert tasting, but rather to identify the volatiles responsible for and precursors of aromas that emanate from the samples and the relationships among them. To address this objective, a study of physicochemical parameters (pH, moisture, and tenderness) was conducted, along with an analysis of volatile compounds using SPME-GC/MS. Finally, a principal component analysis (PCA) and a discriminant analysis (DA) were performed to explore the distribution and grouping of the samples based on various factors.

Detection of antibiotics in pork for the use of new animal welfare technologies during life and at slaughter, ensuring food safety and improving meat quality.

The aim of this project is to improve food quality and ensure safety in the consumption of the product generated by reducing or even eliminating antibiotics in meat products. To achieve this goal, a program will be implemented to enhance animal welfare, boost their immune system, and eliminate stressful situations, thereby reducing the preventive medication administered to the animal. To obtain empirical results, it will be necessary to study antibiotic levels at ´trace´ levels. This will require the development of a sampling protocol and the selection and design of analytical instrumentation to obtain reliable results of these drug levels using a validated equipment for this purpose.

Improvement of food safety through the reduction of nitrifiers and specific control of curing parameters in the ham manufacturing process.

The amount of fat and the weight of the ham are criteria used in the selection of raw materials and determine the processing time of the ham. While in northern European countries, consumers prefer lean hams, in some Mediterranean countries, part of the population accepts a certain amount of infiltrated fat in this product, which allows for an extended maturation period, resulting in sensory characteristics that are highly valued. In Iberian hams and long-cured serrano hams, the presence of infiltrated fat and a certain amount of surface fat slows down the drying process and impregnates the muscle, enhancing the chewing process during consumption and providing a creamy sensation in the mouth along with a highly appreciated aged flavor. The pH of the meat is another important parameter affecting the maturation of the ham. In this regard, most authors recommend avoiding hams with a pH > 6.2 for microbiological safety reasons, to improve salting, to reduce the percentage of spoiled hams, and to avoid issues such as appearance, phosphate precipitates, and soft texture. This issue is more significant in large hams. Furthermore, within the same ham, pH values vary considerably between muscles. To obtain an easy and representative measurement of the ham, it is advisable to take the measurement in a sizable external muscle, such as the semimembranosus.

Temporal evolution of antibiotic levels in pork meat, viscera, and fatty tissues to ensure compliance with legal requirements of countries outside the European Union.

In the production of livestock, a wide variety of pharmacological products are used, whether for therapeutic, zootechnical purposes, or as growth promoters. The vast majority of these products are likely to leave residues in the food derived from treated animals, either in their original active form or as metabolites. The administration of these substances for therapeutic purposes is often necessary, but when used fraudulently, indiscriminately, and abusively, without adhering to the principles of good veterinary practice, the presence of residues in food can pose a serious risk to consumer health. In the context of pig farming for consumption, the presence of pharmacological residues in meat, viscera, and fatty tissues is closely related to the structure and chemical characteristics of each drug, the type of administration, the amount administered, the animal`s lifestyle, interactions with other drugs, and a long etcetera that makes this topic a field of research that has not been thoroughly addressed in its complexity. So far, there are many more statistical studies than empirical ones due to the complexity of combining all the variables in experimental execution. In this work, a study has been conducted on the legal twists, health risks, and the temporal evolution of antibiotics in pork intended for consumption, with a final section dedicated to some aspects related to export regulations to non-EU countries, such as Russia, where there is particular controversy regarding the maximum allowed levels of drug residues in its meat products.

Combination of water activity and pH to improve food safety in microbiological quality outcomes.

The synergy of variables in critical environments allows for the achievement of a highly ambitious goal, which consists of preserving the meat product with minimal alteration so that it retains the desired organoleptic properties, while at the same time ensuring the microbiological safety of the food. This synergy between factors represented a major advance in the development of the food industry, providing solutions that went beyond the state of the art known at that time. Smart critical environment technology has advanced in parallel with the growing knowledge of the behavior of different microorganisms in various situations, including understanding stress conditions in microorganisms, as well as homeostasis and metabolic exhaustion.

Study of livestock feeding for the improvement of food safety in microbiological quality outcomes.

Different studies have confirmed the possibility of increasing the endogenous preservation capacity of meat through the ingestion of by-products from plants rich in active preservative compounds. Dietary supplementation techniques using natural preservatives incorporated into animal feed have proven to be highly effective, as many active compounds are metabolized and deposited in the cell membranes of muscle and adipose tissue, where they carry out their action more effectively. Among the plants studied, aromatic-medicinal plants (AMPs), which are rich in phenolic compounds, have attracted particular interest because their assimilation through the diet of lambs leads to optimal antioxidant and/or antimicrobial activity. In this context, the dietary use of natural oil extracts with specific properties has been shown to be effective in increasing the preservation capacity of meat.

Search for natural alternatives to preservatives and antioxidants that are precursors of nitrosamines.

Given the focus that European directives are placing on the progressive reduction of nitrate and nitrite salts in processed meats, and the increasing awareness of modern society regarding food safety and quality, it is essential to open new lines of research that will allow us to advance in the search for new additives for use in meat products and fresh meat. These new additives should provide a healthier product that can be consumed safely in the long term. For this reason, an alternative to nitrosamine precursor products is being sought through the combination of antibacterial agents and colorants that together can offer meat the same advantages as nitrifying salts, without compromising consumer health. Some natural ingredients, such as plant extracts, offer an alternative to the use of artificial nitrosamine precursor additives in meat products due to their coloring properties. These active ingredients can be incorporated through direct addition or by dietary supplementation in livestock. Endogenous preservatives are deposited in muscle and are highly effective in enhancing the action of preservatives used in product processing. Studies are also being conducted on the use of blocking and inhibiting agents of nitrosamines in meat products to prevent undesirable reactions in the food that could affect the safety of the product.

Machine learning algorithms for the evaluation of risk by tick-borne pathogens in Europe

Two researchers address the problem of building an open-access spatial database as a framework for complex modeling in assessing the risk of tick-borne pathogens in Europe

Novel herpesvirus in the critically endangered Galapagos pink land iguana.

Virus surveillance in wildlife is important to understanding ecosystem health, taxonomy, and evolution. Nevertheless, viruses in reptiles, and specifically in squamates, continue to be understudied. Herein, we conducted a health assessment on the critically endangered Galapagos pink land iguana (Conolophus marthae) and the vulnerable Galapagos land iguana (Conolophus subcristatus). We collected oral and/or cloacal swabs from 47 clinically healthy iguanas and tested for adenovirus (cloacal swabs, n = 47) and herpesvirus (oral swabs, n = 45) using broad-spectrum PCRs. Two out of 38 (5.3 %) Galapagos pink land iguanas tested positive for herpesvirus, while no herpesvirus was detected in all Galapagos land iguanas (n = 7). Both herpesviral sequences were identical between them and divergent (61.9 % amino acid identity) when compared to the closest herpesvirus sequences available in GenBank/EMBL/DDBJ. The genetic distance between this and other herpesviruses is consistent with its classification as a novel virus species. All iguanas were negative for adenovirus. This is the first description of a herpesvirus in iguanas of the Galapagos islands, and the first report of a potential pathogen for the iconic Galapagos pink land iguana. Further research is needed to understand the implications of this virus in the conservation and management of one of the most endangered iguana species in the world.

Campylobacter assessment along the Spanish food chain: identification of key points.

Aims Campylobacteriosis, caused by Campylobacter spp., is one of the most important foodborne zoonotic diseases in the world and a common cause of gastroenteritis. In the European Union, campylobacteriosis is considered the most common zoonotic disease, with over 10,000 cases in 2020 alone. This high occurrence highlights the need of more efficient surveillance methods and identification of key points. Methods and Results Herein, we evaluated and identified key points of Campylobacter spp. occurrence along the Spanish food chain during 2015–2020, based on the following variables: product, stage and region. We analysed a dataset provided by the Spanish Agency for Food Safety and Nutrition using a machine learning algorithm (random forests). Campylobacter presence was influenced by the three selected explanatory variables, especially by product, followed by region and stage. Among the studied products, meat, especially poultry and sheep, presented the highest probability of occurrence of Campylobacter, where the bacterium was present in the initial, intermediate and final stages (e.g., wholesale, retail) of the food chain. The presence in final stages may represent direct consumer exposure to the bacteria. Conclussions By using the random forest method, this study contributes to the identification of Campylobacter key points and the evaluation of control efforts in the Spanish food chain.

Comparative evaluation of pathogenetic mechanisms and immune response in wild boar and domestic pigs experimentally infected with currently circulating African swine fever virus genotype II isolate

Safety and vaccine efficacy of the Rift Valley fever virus 40FP8 variant

Comparative study of pathogenic mechanisms and immune response in sheep infected with Rift Valley virus by intranasal or subcutaneous route.

Veterinary pathology as a useful tool for the development of new vaccines and treatments against respiratory viral pathogens.

The objective of this line of research is focused on the histopathological evaluation and categorization of lesions caused by viral pathogens (SARS-CoV-2, MERS virus, influenza virus) in the respiratory tract in murine models. It also focuses on the development, standardization and implementation of immunohistochemical techniques for the in vivo evaluation of the presence and distribution of these viral pathogens in the respiratory tract and their role in the pathogenic mechanisms of injury. These tools will contribute to the evaluation of the efficacy and safety of new vaccines and treatments.

Evaluation of pathogenic mechanisms and immune response in wild boar and domestic pigs against African swine fever virus.

The objectives of this line of research are to deepen the knowledge of the pathogenic mechanisms and immune response in wild boar and domestic pigs against African swine fever virus (ASFV). Using ancestral Caucasian isolates of ASF virus genotype II that originally circulated after the reintroduction of the disease in Europe in 2007, comparative experimental inoculations will be carried out in domestic pigs and wild boar, as well as time course studies. This will allow a comparative assessment of disease course, clinical signs, macroscopic and histopathological lesions, viraemia levels, viral shedding pathways, viral load in target organs and virus-cell interactions, together with characterization of the systemic and local immune response. The results will contribute to the development of new vaccines to control the spread.

Study of pathogenic mechanisms and immune response against Rift Valley fever virus in livestock.

Understanding the pathogenesis of Rift valley fever virus (RVFV) infection in susceptible hosts is a prerequisite for developing models for testing RVFV vaccines and to identify new targets for clinical interventions. In general, domesticated ruminant models of RVFV have not been studied in detail over the course of infection, so that questions such as initial target organs for virus replications and virus-cells interactions, virus spread mechanisms depending on the route of infection, viremia titers as well as virus burden and target cells in specific tissues remain unclear. Due to the phylogenetic closeness with humans, sheep is a good model for conducting pre-clinical evaluations of vaccines as well as for studying RVFV-associated disease in humans and other livestock since pathological, virological and immunological assessments can easily be performed in this small ruminant highly sensible to RVFV infection.

Conservation of Threatened or Endangered Small Cetaceans Fund

Veterinary epidemiology of infectious diseases.

Our group works in epidemiology and veterinary preventive medicine of emerging and/or zoonotic pathogens in livestock, wildlife, and pets, such as African swine fever viruses, avian influenza, and SARS-CoV-2. We address veterinary health issues from a One Health perspective, combining risk analysis, studies using geographic information systems, spatiotemporal and conventional statistics, data science, and artificial intelligence. This approach allows us to identify the dynamics of diseases and their risk factors, as well as to generate predictive models to improve and focus preparedness for emerging infectious diseases. It also enables us to develop tailored surveillance and control strategies for each epidemiological scenario or ecosystem. Some examples of our work include: Early warning system for the entry of avian influenza in Spain, DiFLUsion. Interactive global map of notifications for African swine fever, avian influenza, and SARS-CoV-2. Cartographic viewer of wild boar distribution in Eurasia.

Epidemiological Tools to Mitigate Pandemic Effects in a Climatic Changing Scenario - LINCGLOBAL 2024

First detection of herpesvirus and hemosporidians in the endangered Pyrenean Capercaillie (Tetrao urogallus aquitanicus).

Pathogens affect wild bird populations worldwide, contributing to their decline. Considering the scarce health data regarding the endangered Pyrenean Capercaillie (Tetrao urogallus aquitanicus), we molecularly surveyed selected pathogens (Newcastle disease virus, Avian influenza virus, Chlamydia psittaci, avian pathogenic Escherichia coli, Campylobacter jejuni, and Salmonella spp.) in 30 Pyrenean Capercaillie feces collected in the field (Catalonia, northeastern Spain). Additionally, swab and tissue samples from eight wild Pyrenean Capercaillies of Catalonia and Andorra were molecularly tested for herpesvirus and hemosporidians (Plasmodium spp., Haemoproteus spp., and Leucocytozoon spp.). All fecal samples were negative for the pathogens tested. Nevertheless, we detected a novel herpesvirus in 50% (4/8) of the Pyrenean Capercaillies, and hemosporidian DNA in 62.5% (5/8) of the tissue samples (i.e., Haemoproteus sp. [4 of 8] and/or Leucocytozoon sp. [3 of 8]). To our knowledge, this is the first detection of herpesvirus and hemosporidians infections in Pyrenean Capercaillies. The putative novel herpesvirus belongs to the genus Iltovirus. The presence of hemosporidian parasites in this mountain bird species is of concern, and could be related to the marked increase in the average temperature in the Pyrenees as a consequence of climate change. Our findings are fundamental to improve the conservation plans for the endangered Pyrenean Capercaillie population.

Molecular diagnostic service for wildlife

Our group has numerous protocols to detect infectious agents of interest in wildlife using molecular biology techniques, such as Plasmodium spp., Haemoproteus spp., Toxoplasma gondii, fungal and bacterial agents, and viral agents (e.g., herpesvirus, adenovirus, poxvirus).

Antimicrobial resistances in the environment.

Antimicrobial resistance is one of the greatest public health challenges of the 21st century. At EySa, we evaluate the role of the environment in the epidemiology of antimicrobial resistances. Our two main lines of study are: - Dissemination of antimicrobial resistances in terrestrial environments and their impact on wildlife and the food chain. - Assessment of the environmental risk of veterinary antimicrobial medications. These approaches are carried out through direct detection of DNA in the environmental resistome and the development of geospatial models of habitat vulnerability to antimicrobials.

Detection and characterization of zoonotic and/or emerging pathogens.

This line of research comprises two main fronts: - Studies on the baseline health status in urban and wild animal populations. - Identification of etiological agents and risk factors in wildlife mortality/morbidity events. Some examples include studies on infectious agents that may impact the conservation of flagship species such as the Pyrenean capercaillie (Tetrao urogallus aquitanicus) and the Iberian wolf (Canis lupus signatus), in collaboration with wildlife rehabilitation centers and national and international institutions (Norwegian Veterinary Institute, Charles Darwin Scientific Station).

Novel epidemiologic features of highly pathogenic avian influenza virus H5N1 2.3.3.4b panzootic: a review.

Avian influenza is one of the most devastating avian diseases. The current high pathogenicity avian influenza (HPAI) A virus H5N1 clade 2.3.4.4b epizootic began in the 2020–2021 season, and has caused a panzootic, considered one of the worst ever reported. The present panzootic has novel epidemiological features that represent a challenge for its prevention and control. This review examines key epidemiological changes of the disease such as seasonality, geographic spread, and host range. The seasonality of the virus has changed, and contrary to previous avian influenza epizootics, this subclade was able to persist during boreal summer. Its geographic range has expanded, with reports in all continents except Australia. During this epizootic, HPAIV H5N1 has broadened its host range, infecting hundreds of bird species, and causing the death of thousands of wild birds and over 300 million poultry. The number and diversity of mammal species infected by H5N1 2.3.4.4b is unprecedented. Although considered low, this strain’s potential to spillover to humans should not be underestimated, especially considering the current extremely high viral circulation in animals and increasing adaptation to mammals. Overall, HPAI A(H5N1) clade 2.3.4.4b represents an ongoing and growing threat to poultry, wildlife, and human health.

Subolesin as anti-tick vaccine for cattle in Uganda

Scientists develop an anti-tick vaccine for cattle in Uganda based on the Subolesin antigen, whose efficacy, effectiveness and safety have been evaluated in a field trial on cattle infested with multiple tick species in different agro-ecological zones of this African country.

Neuroscience zoological perspective

Physiology of the nervous system and Psychobiology of animal behavior

Animal sleep studies from a phylogenetic perspective

Wide-range intelligent system, based on biometric data, for the control of animal behavior and well-being

New evidence for link between tick saliva proteins and lipids and red meat allergy

Recent study provides results on new relationships between different protein and lipid molecules in tick saliva and alpha-Gal syndrome (allergic reaction associated with tick bites and ingestion of red meat) using the model validated in zebrafish

When and where is the Ixodes ricinus tick bite most likely to occur?

New statistical model predicts when and where Ixodes ricinus ticks are most likely to bite people and animals, providing a key tool for preventing pathogen transmission.

Establishing partnerships improves access to livestock biotechnology products in sub-Saharan Africa

Project developed by the SaBio Group of the IREC in collaboration with the National Agricultural Research Organization of Uganda (NARO) for the production of livestock tick vaccines in Uganda values the establishment of collaborations between low-income countries and foreign partners as a strategy to promote the development of livestock biotechnology products adapted to local needs and manufactured in regional facilities.

Knowledge and public perception about wild ungulate populations in Spain

Study shows that the disconnection between public perception and scientific reality on the abundance of wild ungulate populations in Spain, influenced by the media, conditions the social acceptance of population management measures for these species.

Wild boar as sentinel of Crimean-Congo hemorrhagic fever in Spain and Portugal

Epidemiological model based on wild boar allows the identification of new high-risk foci of exposure to the Crimean-Congo hemorrhagic fever virus in Spain and Portugal and will be key to preventing the disease in humans

Ticks and gut microbiota: the connection with allergic reactions to mammalian meat consumption

Study provides new results on the role of gut microbiota in allergic reactions to components of tick saliva using the model of the alpha-Gal syndrome (allergic reaction to ingested red meat) in zebrafish

The impact of myxomatosis on Iberian hare populations

Study analyzes the decline of hare populations in the Iberian Peninsula after the jump of myxoma virus from rabbits using hunting statistics

Optimization of bioactives extraction from grape marc via a medium scale ambient temperature system and stability study

A scalable procedure with minimum energy requirements, in combination with solvents generally recognized as safe (GRAS), has been optimized to obtain polyphenolic extracts from white grape (Vitis vinifera) marc.

Procedure for obtaining and isolating an organic fertilizer and grape seeds from grape residues

Polyphenolic extract from white grape residue

NeoGiANT. The power of grape extracts: antimicrobial and antioxidant properties to prevent the use of antibiotics in farmed animals

Bioactive extracts as antifungal therapies in forest pathology and eubiotic additives in animal nutrition

Extraction and analysis of bioactive compounds in feed and environmental samples

Development and application of sustainable extraction techniques (Green Chemistry) and analysis by liquid/gas chromatography and mass spectrometry to determine polyphenols and other bioactive compounds in feed and environmental samples (water, soil...). Determination of metabolites in samples of animal origin by high resolution mass spectrometry.

Determination of total polyphenol content and antioxidant activity in samples of animal origin or feed

Determination of total polyphenol index (TPC) by Folin method and antioxidant activity (AA) by DPPH and ABTS methods in samples of animal origin and feed

Crimean-Congo hemorrhagic fever virus detected for the first time in free-living ticks in a periurban area of Ponferrada

A study detects the Crimean-Congo hemorrhagic fever virus in 10 of 95 ticks analyzed, none of the genus Hyalomma, captured in a periurban area of Ponferrada. In the region of El Bierzo (León), three clinical cases in humans have been reported since 2021, with one death, due to this disease.

The flight of spotless starlings is related to the bacteria they carry

Combining leading satellite telemetry and epidemiology technology, a study has revealed how the spotless starling (Sturnus unicolor), a common species on livestock farms, connects different farms, urban and natural environments, as well as that the nature of its movements is related to infection by commensal E. coli and antibiotic resistance.

Multi-omics analysis of zebrafish response to tick saliva reveals biological processes associated with alpha-Gal syndrome

A new study used a multi-omics approach to determine the effect of tick saliva and red meat consumption on the transcriptome and gut proteome of zebrafish

New survey to study outbreaks of Epizootic Hemorrhagic Disease in deer

The survey is aimed at managers, guards, veterinarians, engineers, hunters and owners of farms who have had suspected cases of the virus and has the objective of expanding knowledge about the situation of the disease, knowing its impact at the national level and adopting decisions. future for its management.

Efficiency of different capture methods for the tick Hyalomma lusitanicum, vector of the Crimean-Congo hemorrhagic fever virus

A study shows that the efficiency of the most used capture methods for adult stages of the tick Hyalomma lusitanicum varies depending on the habitat where they are used, which is key to more accurately estimate its abundance.

Questionnaire on tick bite and red meat allergy in humans

The IREC SaBio Group launches a questionnaire to collect information on the effects of tick bites and their possible association with red meat allergy, known as Alpha-Gal Syndrome, within the framework of the BIOGAL Project.

Survey on epizootic hemorrhagic disease (EHE) for farmers and veterinarians

The SaBio Group of IREC, GISAZ and ENZOEM launch a virtual survey for veterinarians and livestock farmers whose objective is to study EHE outbreaks in cattle and evaluate the impact of the disease in Spain

LANDINM Project – Landscape Immunity

Camera trapping and population density: why should we estimate movement rates for the population under study?

A study demonstrates the need to estimate movement rates in the animal population under study when camera trapping is used together with methods that do not require the recognition of individuals in the field of wildlife monitoring and population density estimation.

'Junk food' also adds pounds in stork nestlings

The nestlings of storks fed with food from urban landfills are in better nutritional status than those fed with more natural food. However, the effect of food from landfills on the oxidative stress balance largely depends on the climate as this affects nutrient contents of both natural and landfill-derived food.

Allergic reactions to tick saliva components in zebrafish model

Components of tick saliva and proteins with and without alpha-Gal modifications involved in allergy caused by this carbohydrate are identified and characterized.

Channel-based biosensors to support a precision agriculture approach for improved bovine mastitis management (Biosens4PrecisionMastitis)

Detection of antimicrobial resistance

Biosensors for the detection of key antibiotic resistances and of bacterial biofilm growth.

Pathogen detection

Biosensors for pathogen detection, mainly for viruses and bacteria detection.

Antibiotics detection

Biosensors for the detection of antibiotics.

Early diagnosis of infectious diseases through the detection of key host immune-derived biomarkers

Biosensors for the early diagnosis of infectious diseases through the detection of key host immune-derived biomarkers, such as miRNAs, cytokines, etc.

How to deal with an outbreak of African swine fever in wild boar?

A scientific review and consultation with experts reveals the keys to control an outbreak of African swine fever (ASF) in wild boars in Spain

First case of mortality associated with pentobarbital intoxication in a bearded vulture

The case highlights the risk posed by the carcasses of domestic animals euthanized with veterinary pharmaceuticals such as pentobarbital for the conservation of avian scavengers, including the bearded vulture, considered endangered in Europe.

Genetic modifications are introduced into bacteria to control Anaplasma phagocytophilum infection in ticks

IREC researchers have developed modified bacteria using molecular biotechnology that express Anaplasma phagocytophilum proteins capable of competing with this pathogen and reducing infection.

Canine adipose tissue mesenchymal stem cell secretome (CAD-MSCS) and development of its clinical application in veterinary medicine (SecMeV)

Proof of concept to reach TRL 4 for an immunomodulatory product based on human umbilical cord mesenchymal stem cell exosomes, for use in severe cases of Covid-19. Development of in vitro and in vivo tests (EXOMOD)

Preclinical evaluation of personalized, bioprinted, implants for the surgical correction of critical-sized cranio-maxillofacial defects in pediatric patients (FacePrint)

Biomechanical and biological analysis of the combination of osteosynthesis system and scaffold for the repair of segmental bone defects (SEGBONREP)

Advanced prostheses and implants based on 3D printing

Use of 3D printing technology to design novel customised prosthetic implants. Implementation through bioactive coatings and tissue engineering elements (molecules, cells or cell products, resorbable biomaterials)

Cell Therapy, Tissue Engineering, Biomaterials, ATMP

Research on the therapeutic potential of different stem cell types (adult and perinatal SC). Determination of the culture conditions needed to use SC as therapeutic product per sé, or as part of a tissue engineering product. Research on the therapeutic potential of stem cell products (exosomes, secretome...). Development and evaluation of biomaterials to be used as tissue substitutes (mainly bone) in reconstructive surgery, or as a scaffold in tissue engineering. Our goal is developing products for humans, but since preclinical evaluation takes place in animals (rat, rabbit, minipig), we have sometimes collaborated with veterinarians, finding novel applications to our research.

Riesgo de exposición al virus de la fiebre del Nilo Occidental en aves silvestres, caballos y personas del oeste de Castilla-La Mancha

Las aves silvestres de entornos rurales humanizados como las granjas de caballos en el occidente de Castilla-La Mancha son más proclives a estar infectadas por Flavivirus como el virus de la fiebre del Nilo Occidental, representando un riesgo para la transmisión del virus a caballos y personas en estos ambientes.

Viral-environmental epidemiology

Monitoring of natural and anthropogenic environments for disease surveillance, especially viral outbreaks.

Biomedical and biotechnological applications of phages

Development of bacteriophages as alternative biomedical tools in the fight against multi-resistant bacteria, including their use in diagnosis, prevention and therapy. Isolation and characterisation of new bacteriophages, directed evolution for their optimisation, emergence of resistance and nanotechnology.

Nanobiotechnology applied to the manufacture of DNA vaccines

We have designed & patented silica-based physiologically responsive particles to encapsulate, store, and transfer DNA. Unlike existing vectors (e.g., viral or lipidic particles), these DNA@SiO2 systems are very stable at room temperature. We also demonstrate how they protect the encapsulated DNA from exposure to different biological and physicochemical stresses, including DNase, denaturation temperatures (>100 °C), or reactive oxygen species (ROS). Remarkably, upon cellular uptake, these vectors dissolve safely unpacking the DNA and transfecting the cells. The versatility of the design is such that it can encapsulate genes without gene/size restrictions, in single or multiple layers of silica, so different genes can be expressed sequentially. This allows the time-controlled transcription of several genes, mimicking viral gene expression cascades, or even ´fine-tuning´ gene expression in transfected cells on demand. In addition, the method is easily scalable, reproducible, and inexpensive, enabling large-scale production and batch-quality testing, all of which are important for the personalized therapeutics industry. The high stability of these DNA vectors allows for easy and low-cost transport from the point of production to virtually any destination, making them unique as gene delivery tools. More info here: https://www.sciencedirect.com/science/article/pii/S2590049823000176/pdfft?md5=ca0c16339faf79dc7061b3940ac98989&pid=1-s2.0-S2590049823000176-main.pdf

See updated list here: https://mlfanarraga.wixsite.com/grupo-nanomedicina/recent-publications

https://mlfanarraga.wixsite.com/grupo-nanomedicina/recent-publications

Silica Particles for Nucleic Acid Encapsulation

Procedure for the identification of compounds that inhibit the polymerization process of phfs-type tau filaments and their applications

Composition of nanofilaments for the treatment of tumors

Method of protein coating of materials by electrostatic bioconjugation

Development of encapsulation systems for medicines based on solid lipid particles

The encapsulation system based on nanoparticles of a lipid nature serves to change the bio-distribution of any drug, thus modifying its tissue concentrations, targeting and toxicity. More info: https://www.mdpi.com/1999-4923/13/1/93/pdf?version=1610543511

Design of a nano-biosensors for the screening and rapid detection of different pathologies

Different types of immunoassays to measure the presence or concentration of analytes in biological fluids are based on the ability of an antibody to recognize and bind to a specific analyte in a complex mixture of macromolecules. Nanoparticle-based biodetection tests exhibit interesting properties including: increased sensitivity, specificity, portability, and versatility for the detection of a variety of biomarkers. Lateral flow assays (LFAs), also known as ´lateral flow tests´ or ´lateral flow immunoassays´ represent one of the fastest growing qualitative and quantitative immunoassays that can be used for the detection of many different antigens for the early detection of diseases and monitoring of patients. LFA assays have long been used to determine pregnancy, but are now synonymous with COVID-19 (SARS-CoV-2) antigen tests. These tests are highly specific, robust, low-cost, require a small amount of sample volume, are fast, and have an easy-to-use assay format. They can be qualitative or quantitative, combined with reading technology. In addition, its development and operating costs are very low compared to other diagnostic methods such as molecular profiling, and due to its versatile nature, it could also be used to detect cancer as a liquid biopsy test.

NOTA DIVULGATIVA: “Las garrapatas y la alergia a la carne roja”

Las garrapatas son ectoparásitos artrópodos que actúan como vectores de patógenos y causan reacciones alérgicas, afectando a la salud humana y animal en todo el mundo. En los seres humanos, sus picaduras pueden inducir altos niveles de anticuerpos IgE contra el carbohidrato alfa-Gal presente en biomoléculas de su saliva, que median reacciones alérgicas a sus picaduras y a la carne de mamíferos conocidas como síndrome alfa-Gal. Identificar y caracterizar biomoléculas derivadas de garrapatas como alfa-Gal y mecanismos inmunes asociados ligados al desarrollo del síndrome de ?-Gal es esencial en el diagnóstico y la prevención de enfermedades causadas por patógenos portadores de alfa-Gal que puedan convertirse en pandémicas.

Tuberculosis is detected for the first time in Egyptian mongooses in Spain

Researchers suggest that the Egyptian mongoose could act as an accidental host for tuberculosis, but that it would play a minor role in the maintenance and transmission of the disease in ecosystems.

Mapeo de epítopos de la proteína RBD Spike del SARS-CoV-2 asociados con sintomatología y estrategias de control de la enfermedad COVID-19

Se identifican epítopos de la proteína RBD del SARS-CoV-2 reconocidos por diferentes isotipos de anticuerpos para avanzar en el desarrollo de intervenciones diagnósticas, pronósticas y preventivas mediante mapeo peptídico eficaces de la enfermedad COVID-19.

Synthesis and biological study of substituted 3-carboethoxy-4-aminoquinolines

Resume: It synthesizes a series of 3-carboethoxy-4-aminoquinolines substituted in position 6 and another series substituted in position 7. The study by molecular modeling makes it possible to determine the molecular descriptors that appear to be important in biological activity. Most of the compounds were cytotoxicly innocuous at the concentration of 50mg/ml, which makes them attractive for further studies.

Molecular Parameters and Structure Activity Relationship towards Mexican Leishmania of 3-Carboethoxy-4-Amino Quinolines.

Resume Some quinolines and related compounds are known to be active as antiparasitic, antihypertensive, and antitumor agents. One of the proposed mechanisms is the intercalative interaction or not with DNA segments. This interaction has been demonstrated by NMR, UV-visible and computational methods. This work presents a computational study to elucidate essential structural elements for pharmacological activity, in a set of compounds that includes known antimalarial drugs: 2,4-diamino-6,7-dimethoxyquinoline (DDQ), amodiaquine, chloroquine, and a series of 3-carboethoxy-4-N-alkylaminoquinolines synthesized in our laboratories and whose biological activity towards Lehismania mexicana we determined. Energetically accessible conformations of the series of compounds were generated and geometrically optimized using molecular dynamics (MM+) and minimization (simulated annealing) and later optimized for the calculation of electronic properties using the AM1 semi-empirical method. Molecular properties were calculated including electrostatic potential maps, HOMO-LUMO energies, dipole moments, heats of formation, charge density (Mülliken), solvation energy, octanol-water partition coefficient (log P), polarizability, hardness, surface area. , volume and thermodynamic parameters, using the AM1 Hamiltonian model in order to identify properties that can be correlated with biological activity. Pharmacophore models are proposed based on the identification of common structural patterns, both for antimalarial drugs and for the group of 3-carboethoxy-4-N-alkylaminoquinolines whose activity against Lehismania mexicana was determined. The proposed pharmacophore models will serve as the basis for molecular design in the search for new drugs with the desired activity.

Fish welfare

Current research in IRTA focusses on fish stunning methods and evaluate consciousness in fish. We are also developing and validating operational welfare indicators in fish to set up and implement a fish welfare certification scheme.

Virus-Host Interaction

We investigate the interactions of animal and human highly pathogenic viruses with the host they infect, and the innate immune response, with the aim of identifying crucial cellular components in the infection that will be used as targets to develop new vaccine or antiviral strategies. For this purpose, we use high-throughput, proteomics and trancriptomics.https://www.inia.es/en-en/Research/Animalresearch/Biotechnology/Virus-Host-Interactions/Pages/Home.aspx

Identifican biomarcadores nutricionales del pronóstico de la COVID-19

Un nuevo estudio identifica biomarcadores nutricionales del pronóstico de la enfermedad COVID-19 en suero en individuos no vacunados y tras la administración de la vacuna

La monitorización integrada de fauna silvestre como herramienta para la prevención de futuras pandemias

La monitorización integrada de fauna silvestre, que nace del concepto One Health y propone combinar la vigilancia sanitaria con el seguimiento de las poblaciones animales, ha sido ensayada por primera vez en España como herramienta para prevenir futuras pandemias.

Abundance of the tick Hyalomma lusitanicum determines the risk of transmission of the Crimean-Congo hemorrhagic fever virus

This study shows that the abundance of Hyalomma lusitanicum at small spatial scales and the structure of the community of its hosts, mainly deer and wild boar, are important risk factors for the transmission of the Crimean-Congo haemorrhagic fever virus.

The IREC receives the One Health 2022 Recognition from Boehringer Ingelheim

The Boehringer Ingelheim One Health 2022 Recognition aims to support and make visible those initiatives that make, through their daily activity, the advancement of solutions for human, animal and environmental well-being a reality.

Air purifiers and SARS-CoV-2: HEPA filter analysis as a surveillance method

Researchers from Castilla-La Mancha lead the development of a method for detecting the SARS-CoV-2 virus in safe environments, based on the analysis of HEPA filters contained in air purifiers, which would reduce the economic costs of individual analysis and speed up the diagnostic process of COVID-19 infection.

Environmental determinants of the local abundance of ticks in Doñana National Park.

New models in high spatial resolution allow predicting the abundance of ticks and improve the prevention of the serious diseases they transmit.

New Bagaza virus outbreak in red-legged partridges in Spain and appearance in Portugal and other birds.

Two studies analyze the Bagaza virus outbreaks in birds that took place for the second time in Southern Spain in 2019 and for the first time in Portugal in 2021. The data confirm that co-infection of this flavivirus and avian malaria parasites is the cause of the high mortality observed in the affected populations of red-legged partridge.

cardiac fibrosis

Study of the molecular basis of fibrotic pathology and the molecular mechanisms related to TGFβ and Hsp90 for their treatment in vitro and in vivo. Study the effects of new pharmacological strategies based on biotechnology, CRISPR/Cas mutations or modified plasmids for their use as teragnostic particles in vitro and in vivo. Study of antifibrotic therapy and telomeric protection against SARS-CoV2 infection. Generation of new nano-transport systems to reach therapeutic targets. Generation of new diagnostic tools.

Modular equipment for in vitro digestion

Urban pollutants and SARS-CoV-2 virus

A multidisciplinary research reveals that atmospheric particulate matter emitted by engine exhaust inactivates SARS-CoV-2, but it has a negative impact on human health with implications for COVID-19 and other diseases

Hunting yields to predict wild boar abundance

A study demonstrates the predictive capacity of spatial models of wild boar relative abundance, generated in 2014 from data from hunting yields, comparing them with current hunting yields of the species.

The snow-melt mosquito, Aedes refiki, colonizes Ciudad Real.

Several specimens of a very aggressive mosquito that usually lives in cold high mountain environments are captured in the Forest Park “La Atalaya”, in the city of Ciudad Real.

The REM: a reliable method for estimating population density of multiple wildlife species using camera traps

The Random Encounter Model (REM), based on the use of camera traps, has been validated as a reliable method for estimating population density of multiple wildlife species with the same study design.

New oral vaccines to help food production in Africa

The SaBio Group of the IREC has successfully tested an oral vaccine that combines tick antigens with inactivated mycobacteria, and that is expected to help control cattle ticks and reduce the use of chemical acaricides in tropical and subtropical countries.

Can we model distribution of population abundance from wildlife–vehicles collision data?

A new analytical approach makes it possible to obtain maps on the distribution and abundance of two species of ungulates, the roe deer and the wild boar, from wildlife–vehicles collision data, controlling the effects that factors such as road density and seasonality may have on predictions.

The common mosquito transmits many diseases, but is unable to transmit hepatitis E virus

A study shows that the common mosquito is unable to transmit the hepatitis E virus, an emerging virus that causes severe cases of acute hepatitis and that causes the death of tens of thousands of people worldwide.

New benzophenone-derived bisphosphonium salts as leishmaniccidal leads targeting mitochondria through inhibition of respiratory complex II

A set. of benzophenone-derived bisphosphonium salts was synthesized and assayed for lethal activity on the human protozoan parasite Leishmania. A subset of them, mostly characterized by phosphonium substituents with an intermediate hydrophobicity, inhibited parasite proliferation at low micromolar range of concentrations. The best of this subset, 4, 4`̌prime;-bis((tri-n- pentylphosphonium)methyl)benzophenone dibromide, showed a very scarce toxicity on mammalian, cells. This compound targets complex II of the respiratory chain, of the parasite, based on, (i) a dramatically swollen mitochondrion in treated parasites, (ii) fast decrease of cytoplasmic ATP, (iii) a decrease of the electrochemical mitochondrial potential, and, (iv) inhibition of the oxygen consumption rate using succinate as substrate. Thus, this type of compounds represents a new lead in the development of leishmanicidal drugs.

Antiprotozoal activity and DNA binding of dicationic acridones

Two series of N-alkyl, N-alkoxy, and N-hydroxy bisguanidines derived from the N-phenylbenzamide and 1,3-diphenylurea scaffolds were synthesised in three steps from the corresponding 4-amino-N-(4-aminophenyl)benzamide and 1,3-bis(4-aminophenyl)urea, respectively. All of the new compounds were evaluated in vitro against T. b. rhodesiense (STIB900) trypomastigotes and Plasmodium falciparum NF54 parasites (erythrocytic stage). N-alkoxy and N-hydroxy derivatives showed weak micromolar range IC50 values against T. b. rhodesiense and P. falciparum whereas the N-alkyl analogues displayed submicromolar and low nanomolar IC50 values against P. falciparum and Trypanosoma brucei, respectively. Two compounds, 4-(2-ethylguanidino)-N-(4-(2-ethylguanidino)phenyl)benzamide dihydrochloride (7b) and 4-(2-isopropylguanidino)-N-(4-(2-isopropylguanidino)phenyl)benzamide dihydrochloride (7c), which showed favourable drug-like properties and in vivo efficacy (100% cures) in the STIB900 mouse model of acute human African trypanosomiasis represent interesting leads for further in vivo studies. The binding of these compounds to AT-rich DNA was confirmed by surface plasmon resonance (SPR) biosensor experiments.

Alternative oxidase inhibitors: Mitochondrion-targeting as a strategy for new drugs against pathogenic parasites and fungi

The alternative oxidase (AOX) is a ubiquitous terminal oxidase of plants and many fungi, catalyzing the four-electron reduction of oxygen to water alongside the cytochrome-based electron transfer chain. Unlike the classical electron transfer chain, however, the activity of AOX does not generate adenosine triphosphate but has functions such as thermogenesis and stress response. As it lacks a mammalian counterpart, it has been investigated intensely in pathogenic fungi. However, it is in African trypanosomes, which lack cytochrome-based respiration in their infective stages, that trypanosome alternative oxidase (TAO) plays the central and essential role in their energy metabolism. TAO was validated as a drug target decades ago and among the first inhibitors to be identified was salicylhydroxamic acid (SHAM), which produced the expected trypanocidal effects, especially when potentiated by coadministration with glycerol to inhibit anaerobic energy metabolism as well. However, the efficacy of this combination was too low to be of practical clinical use. The antibiotic ascofuranone (AF) proved a much stronger TAO inhibitor and was able to cure Trypanosoma vivax infections in mice without glycerol and at much lower doses, providing an important proof of concept milestone. Systematic efforts to improve the SHAM and AF scaffolds, aided with the elucidation of the TAO crystal structure, provided detailed structure-activity relationship information and reinvigorated the drug discovery effort. Recently, the coupling of mitochondrion-targeting lipophilic cations to TAO inhibitors has dramatically improved drug targeting and trypanocidal activity while retaining target protein potency. These developments appear to have finally signposted the way to preclinical development of TAO inhibitors.

Synthesis, biological, and photophysical studies of molecular rotor-based fluorescent inhibitors of the Trypanosome Alternative Oxidase

We have recently reported on the development and trypanocidal activity of a class of inhibitors of Trypanosome Alternative Oxidase (TAO) that are targeted to the mitochondrial matrix by coupling to lipophilic cations via C14 linkers to enable optimal interaction with the enzyme’s active site. This strategy resulted in a much-enhanced anti-parasite effect, which we ascribed to the greater accumulation of the compound at the location of the target protein, i.e. the mitochondrion, but to date this localization has not been formally established. We therefore synthesized a series of fluorescent analogues to visualize accumulation and distribution within the cell. The fluorophore chosen, julolidine, has the remarkable extra feature of being able to function as a viscosity sensor and might thus additionally act as a probe of the cellular glycerol that is expected to be produced when TAO is inhibited. Two series of fluorescent inhibitor conjugates incorporating a cationic julolidine-based viscosity sensor were synthesized and their photophysical and biological properties were studied. These probes display a red emission, with a high signal-to-noise ratio (SNR), using both single- and two-photon excitation. Upon incubation with T. brucei and mammalian cells, the fluorescent inhibitors 1a and 2a were taken up selectively in the mitochondria as shown by live-cell imaging. Efficient partition of 1a in functional isolated (rat liver) mitochondria was estimated to 66 ± 20% of the total. The compounds inhibited recombinant TAO enzyme in the submicromolar (1a, 2c, 2d) to low nanomolar range (2a) and were effective against WT and multidrug-resistant trypanosome strains (B48, AQP1-3 KO) in the submicromolar range. Good selectivity (SI > 29) over mammalian HEK cells was observed.

Development of drugs targeting mitochondria-like organelles as therapeutic approach to the treatment of neglected parasitic diseases

Compounds derived from 4-amino-N-(4-aminophenyl)benzamide and analogs and use of

Characterisation and management of interaction risks between livestock and wild ungulates in extensive farms in Spain

Scientists from the IREC, the Universities of Cordoba and Leon, together with the research centre VISAVET and the valuable help of collaborators of the sector and administrations, are studying how to characterise and improve biosecurity against wild ungulates through a common and systematic protocol in extensive farms in Mediterranean dehesas.

Inactivated tuberculosis mycobacteria as an immunostimulant against salmonellosis in pigs

Tuberculosis against salmonellosis: A study shows that exposure to inactivated mycobacteria have positive clinical, pathological and immunological effects against other bacterial pathogens in pigs

Use of simple acyclic polyamines for the treatment of diseases caused by Leishmania parasites.

Pharmaceutical and transformation products during unplanned water reuse: Insights into natural attenuation, plant uptake and human health impact under field conditions

In urban and periurban areas, agricultural soils are often irrigated with surface water containing a complex mixture of contaminants due to WWTP effluent discharges. The unplanned water reuse of these resources for crop irrigation can represent a pathway for contaminant propagation and a potential health risk due to their introduction in the food chain. The aim of this study is to provide data about the magnitude of attenuation processes and plant uptake. Target compounds are 25 pharmaceuticals, including transformation products (TPs). The field site is an agricultural parcel where maize is irrigated by a gravity-fed surface system supplied by the Jarama river, a water course strongly impacted by WWTP effluents. Throughout the 3-month irrigation period, irrigation water and water infiltrating through the vadose zone were sampled. The agricultural soil was collected before and after the irrigation campaign, and maize was sampled before harvesting. All selected compounds are detected in irrigation water (up to 12,867 ng L−1). Metformin, two metamizole TPs and valsartan occur with the highest concentrations. For most pharmaceuticals, results demonstrate a high natural attenuation during soil infiltration (>60%). However, leached concentrations of some compounds can be still at concern level (>400 ng L−1). A persistent behavior is observed for carbamazepine, carbamazepine epoxide and sulfamethoxazole. Pharmaceutical soil contents are in the order of ng g−1 and positively ionized compounds accumulate more effectively. Results also indicate the presence of a constant pool of drugs in soils. Only neutral and cationic pharmaceuticals are taken up in maize tissues, mainly in the roots. There is an insignificant threat to human health derived from maize consumption however, additional toxicity tests are recommended for 4AAA and acetaminophen.

Pharmaceuticals and trace metals in the surface water used for crop irrigation: Risk to health or natural attenuation?

The use of surface water impacted by wastewater treatment plant (WWTP) effluents for crop irrigation is a form of unplanned water reuse. Natural attenuation processes can buffer contamination spreading. However, this practice can promote the exposure of crops to contaminants of emerging concern, such as pharmaceuticals, trace metals (TMs) and metalloids, posing a risk to health. This research aimed to evaluate the presence of 50 pharmaceuticals, some transformation products, 7 TMs and a metalloid in the water-sediment-soil-plant system, and their potential to be bioaccumulated into edible parts of plants, as a result of the unplanned water reuse. The study site consists of an extensive agricultural land downstream Madrid city (Spain) where surface water, strongly impacted by WWTP effluents, is applied through gravity-based systems to cultivate mainly maize. Sampling campaigns were conducted to collect WWTP effluent, surface and irrigation water, river sediments, agricultural soils and maize fruits. Results demonstrate the ubiquitous presence of several pharmaceuticals. The concentration pattern in irrigation water did not resemble the pattern of contents in soils and plants. The pharmaceuticals included in the EU surface water watch lists were quantified in the lowest concentration range (antibiotics) or were not detected (hormones). TMs in water and agricultural soils should not arise any concern. Whereas, their presence in the river sediments may have an adverse impact on aquatic ecosystems. Only acetaminophen, ibuprofen, carbamazepine, nicotine, Zn, Cu and Ni were quantified in corn grains. Results indicate that neither pharmaceuticals nor TMs pose a threat to human health due to consumption of maize cultivated in the area.

Antibiotics, hormones, persistent and mobile organic contaminants and pathogens, the complex mixture in agriculture and livestock scenario. Risk to health or natural attenuation? (Nat4Health)

Generating Biomass with Regenerated Waters: Opportunity for Circular Bioeconomy (BioArBio)

Quantification of micronanoplastics in reclaimed water and agricultural ecosystems. Environmental risk assessment (μNanoCare)

Plastic in Agricultural Production: Impacts, Lifecycles and Long-term Sustainability (Papillons)

Discovery and Pharmacological Studies of 4-Hydroxyphenyl-Derived Phosphonium Salts Active in a Mouse Model of Visceral Leishmaniasis

We report the discovery of new 4-hydroxyphenyl phosphonium salt derivatives active in the submicromolar range (EC50 from 0.04 to 0.28 μM, SI > 10) against the protozoan parasite Leishmania donovani. The pharmacokinetics and in vivo oral efficacy of compound 1 [(16-(2,4-dihydroxyphenyl)-16 - oxohexadec yl)triphenylphosphonium bromide] in a mouse model of visceral leishmaniasis were established. Compound 1 reduced the parasite load in spleen (98.9%) and liver (95.3%) of infected mice after an oral dosage of four daily doses of 1.5 mg/kg. Mode of action studies showed that compound 1 diffuses across the plasma membrane, as designed, and targets the mitochondrion of Leishmania parasites. Disruption of the energetic metabolism, with a decrease of intracellular ATP levels as well as mitochondrial depolarization together with a significant reactive oxygen species production, contributes to the leishmanicidal effect of 1. Importantly, this compound was equally effective against antimonials and miltefosine-resistant clinical isolates of Leishmania infantum, indicating its potential as antileishmanial lead.

4-hydroxyphenylphosphonium salts with antiparasitic properties

Antiparasitic chemotherapy of zoonoses

Our group develops the research line ´New chemical entities for drug discovery´ in the field of parasitic diseases, especially those produced by pathogenic protozoa. Since its creation, the group has developed several series of compounds that are highly active against various parasitic protozoa (T. cruzi, T. brucei spp., T. vaginalis, Leishmania spp., Plasmodium spp.). Several lead compounds also demonstrated in vivo efficacy in mouse models of trypanosomiasis, leishmaniasis, and malaria. At present, the group continues the study of these lead compounds, as well as the development of new ones, designed to attack specific targets of these parasites. The groups is primarily working on the design and synthesis of new active molecules. We collaborate with groups from the CSIC or other institutions in Spain and abroad (computational chemists, biochemists, parasitologists, pharmaceutical technologists, crystallographers) to achieve the proposed objectives.

Recent publications list C. Alonso

Recent publications list C. Alonso with links

New insights into the role of endosomal proteins for African swine fever virus infection

African swine fever virus (ASFV) causes a deadly disease of pigs and wild boars that was endemic in Africa but has spread in recent years to Europe, Asia, and Oceania with a high socio-economic impact. ASFV enters the cell by endocytosis and has adapted to the endosomal conditions to acquire infectivity. Fusion of the internal viral membrane with the endosomal membrane is required for the exit of viral DNA into the cytoplasm to start replication. We have found that ASF virion internal membrane proteins E248R and E199L interact with the endosomal proteins Niemann Pick C1 (NPC1) and lysosomal membrane protein (Lamp)-1 and -2. And, appear to be required for endosomal trafficking of ASF virions endosomal traffic and exit to the cytoplasm in the cell entry process. These molecules act regulating cholesterol flux from the endosome to the endoplasmic reticulum and appear to be important for the viral infection cycle. In silenced and knockout cells, ASFV infection was affected at early and later stages. In null cells, virion entry and progression through the endosomal pathway at entry were arrested and several viral cores were retained at late endosomes without entering the fusion phase for the cytoplasmic exit. These results provide new insights into the role of endosomal proteins for ASFV infection

Sulfated phenylhydrazide derivatives as antiviral agents

Sulfur compounds derived from phenylhydrazides as antiviral agents

Do all camera trap models work the same?

Comparing the performance of 45 camera traps of 5 different models under field conditions reveals significant differences based on many factors, such as camera model, deployment height, period of the day or target wildlife species.

First national risk map of Crimean-Congo haemorrhagic fever virus infection.

An epidemiological model based on red deer makes it possible to develop the first risk map of exposure to the Crimean-Congo hemorrhagic fever virus in Spain and will be key to prevent the disease in people.

ORGANOID CULTURE SYSTEM AND METHOD FOR STERILISING AN ORGANOID CULTURE SYSTEM

METHOD FOR PRODUCING GLUCOSE-6-PHOSPHATASE 2 PROTEIN

PROTEIN MUNS THAT CAN FORM INCLUSIONS IN THE ENDOPLASMIC RETICULUM, METHODS FOR THE USE THEREOF AND USES OF SAME

APPLICATIONS OF THE PROTEIN MUNS ANO THE DERIVA TES THEREOF

PEPTIDE WITH QUORUM-SENSING INHIBITORY ACTIVITY, POLYNUCLEOTIDE THAT ENCODES SAID PEPTIDE, AND THE USES THEREOF

Immunization With Bovine Herpesvirus-4-Based Vector Delivering PPRV-H Protein Protects Sheep From PPRV Challenge

The Morbillivirus peste des petits ruminants virus (PPRV) is the causal agent of a highly contagious disease that mostly affects sheep and goats and produces considerable losses in developing countries. Current PPRV control strategies rely on live-attenuated vaccines, which are not ideal, as they cannot differentiate infected from vaccinated animals (DIVA). Recombinant vector-based vaccines expressing viral subunits can provide an alternative to conventional vaccines, as they can be easily paired with DIVA diagnostic tools. In the present work, we used the bovine herpesvirus-4-based vector (BoHV-4-A) to deliver PPRV hemagglutinin H antigen (BoHV-4-A-PPRV-H-ΔTK). Vaccination with BoHV-4-A-PPRV-H-ΔTK protected sheep from virulent PPRV challenge and prevented virus shedding. Protection correlated with anti-PPRV IgGs, neutralizing antibodies and IFN-γ-producing cells induced by the vaccine. Detection of antibodies exclusively against H-PPRV in animal sera and not against other PPRV viral proteins such as F or N could serve as a DIVA diagnostic test when using BoHV-4-A-PPRV-H-ΔTK as vaccine. Our data indicate that BoHV-4-A-PPRV-H-ΔTK could be a promising new approach for PPRV eradication programs.

New vaccine strategies based on recombinant vector

In our group, we are working on developing new vaccine strategies using recombinant viral vectoes such as adenovirus. These vectors are used to express antigens from viral disease such as peste des petits ruminants or bluetongue. The immune response induced by these vaccines is assessed in animal models and in sheep. The most promising constructs are then tested in challenge experiments to determine their protective capacity.

Vaccinomics: a future avenue for vaccine development against emerging pathogens

Scientists from the IREC propose the application of quantum vaccinomics to further contribute to vaccine development by facilitating antigen combinations to target pathogen infection and transmission in current and emerging infectious diseases such as COVID-19.

Identification and characterization of tick salivary biomolecules and mechanisms associated with the immune response to alpha-Gal (BIOGAL)

Pelayo and zebrafish helping in rare diseases research https://cutt.ly/UnT9F5A

Composition for use in mammals, for use in prophylactic and/or therapeutic vaccination against Leishmania

Protein-based nanomaterials as substitutes of antibiotics in animal medicine

Dr. Neus Ferrer is developing protein-based nanomaterials as substitutes of antibiotics in animal medicine. The dry period of dairy cows is addressed to achieve a cellular regeneration of the mammary gland aiming at optimize milk production in the subsequent lactation. The preventive use of antibiotics in this period has become questionable. Therefore, the aim of the project is to develop non-antibiotic preventive alternatives based on the administration of protein-only nanoparticles or encapsulated proteins to improve cow dry period tissue regeneration and immunomodulation. The project is performed in collaboration with Dr. Elena García Fruitós and Anna Arís from the IRTA More information: https://ibb.uab.cat/wp-content/themes/viral/modules/ibb_membres/view_grup.php?CodiGrup=33

Comparative Molecular Physiology

Our primary interest focuses on the molecular basis of gamete (germ cell) formation and function towards the development of biotechnological inventions for animal production and conservation biology. Present research lines include: (1) Comparative studies on the evolution, structure and function of molecular water channels (aquaporins); (2) Molecular physiology of aquaporins and ion channels in male and female gametes; (3) Development of new biotechnological methods based on aquaporins for cell preservation; and (4) Molecular endocrinology of spermiogenesis. Research Keywords: Spermatogenesis, oogenesis, endocrinology, aquaporin, physiology, evolution, cell preservation More info: https://ibb.uab.cat/wp-content/themes/viral/modules/ibb_membres/view_grup.php?CodiGrup=28

Genome Integrity and Instability

The research activity of the research group has as a main objective to study of the mechanism(s) that are responsible for the origin and maintenance of mammalian genome integrity. We reach our main goal through a multidisciplinary approach, combining computational analysis and whole-genome comparisons with cutting-edge experimental technologies in both somatic and meiotic cells. More specifically, the group is currently working in the following research lines: 1. Investigate the conservation and functionality of the high-structural organization of mammalian genomes, both in the somatic and the germ line. 2. Analysis of the signalling pathway that controls the progression of meiotic recombination in mammalian meiocytes. 3. Identification of the role of the DNA damage response machinery in the DSB repair occurring during the meiotic prophase. 4. Study how the DNA damage response mechanism controls the oocyte pool in mammals. 5. Identification of non-annotated genes in the mammalian genome required to complete meiosis. 6. Identification of the genetic basis of reproductive isolation and barriers of gene flow in mammalian natural populations. 7. Development of a cell line repository of endangered mammalian species. 8. Implementation of integrative bioinformatics and informatic tools for the analysis of the conservation and function of vertebrate genomes. 9. Study the impact of COVID-19 on fertility. More information: https://ibb.uab.cat/wp-content/themes/viral/modules/ibb_membres/view_grup.php?CodiGrup=27

Evolutive Immunology

Using a combination of molecular, in vitro and in vivo methodologies we aim to understand host-pathogen interactions and how we can modulate the host immune system to have a good performance against pathogens. We develop our research using zebrafish as a model organism due to its high versatility and the availability of mutants. We do both basic and translational research on fish immunology in three main areas: (1) development of vaccines for animal health, (2) evolution of pathogen recognition in vertebrates and (3) development of diagnostic tools. More info at: https://ibb.uab.cat/wp-content/themes/viral/modules/ibb_membres/view_grup.php?CodiGrup=46

Methods and compositions for controlling or reducing pests

How does feeding in landfills affect white storks?

A multidisciplinary study reveals that white stork nestlings fed with a higher proportion of food from urban landfills present a better nutritional status and body condition than those fed with a higher proportion of natural diet.

Prebiotic composition for the preventative treatment of intestinal infections in livestock

Hunters from Salamanca collaborate with the IREC in the fight against the Crimean-Congo hemorrhagic fever virus

The Salamanca Delegation of the Federación de Caza de Castilla y León collaborates with the IREC in the collection of samples whose analysis will allow elaborating an epidemiological map of the Crimean-Congo virus disease in the province.

Tichinella

For more than 30 years, we have been working on the Trichinellosis research line, both in domestic pigs (white and Iberian) and in wild boars. Scientific studies have focused mainly on both epidemiology and the diagnosis and prophylaxis of this important parasitic zoonosis.

Analysis of the overabundance scenarios of wild ungulates in Europe

A scientific review describes the main scenarios of wild ungulate overabundance that currently exist in Europe and its causes, as well as the most suitable set of indicators of ecological change for its monitoring, diagnosis and management.

An innovative method to detect the presence of Covid-19 RNA in the environment

Developed by a team of nurses, biologists, physicians and veterinarians, and even a physicist, in a true commitment to the ´One Health´ concept to fight the Covid-19, the method will allow us to improve the early detection of the virus, to monitor its circulation and to identify targets for a more efficient control.

Feeding of white storks on landfills could make these vectors for the dispersal of antibiotic resistant bacteria

A multidisciplinary team of researchers reveals that, when foraging on landfills, White storks acquire multiresistant strains of the common bacteria Escherichia coli, and could disperse these over long distances.

Distribution and genetic variability of Crimean-Congo haemorrhagic fever virus in southwestern Spain

Crimean-Congo haemorrhagic fever virus is detected in ticks from wild ungulates from different provinces in southwestern Spain. Its wide distribution and high genetic variability could lead to the appearance of new strains of the virus in the future, with the consequent epidemiological risk.

MYCOTRAINING Project: Nonspecific immunity associated with inactivated mycobacteria: range of pathogens, specificity of the immunogen and molecular mechanisms involved

GOSTU Proyect: Inter-regional Operational Group Tuberculosis

Pilot project to improve biosecurity in extensive cattle farming in the Alcudia Valley

Integrated wildlife disease surveillance - Agreement to carry out tasks related to wildlife health management in Spain

Health and Biotechnology

Specialized diagnosis: pathology - serology - microbiology - new genetic tools. New techniques for comprehensive hunting management. Integrated control of shared diseases. Biotechnology applied to livestock and hunting. Development and application of the latest technologies in integrated wildlife monitoring: censuses, health surveillance, etc. Identification of new antigens and vaccine development. Applications of proteomics, metabolomics and systems biology in the health field and in animal production. Development of new tools for diagnosing infections in wildlife. Population genetics and its application to the solution of health and management needs.

Analytical epidemiology

Epidemiology and control of shared infections such as tuberculosis, African swine fever, vector-borne infections and infections of wild birds; Pathology.

Immunology

Molecular biology of infectious diseases; Human neurological and allergic infectious diseases; Vaccines for the control of vector infestations and transmission of pathogens.

Epidemiology, ecology and evolution of arthropod vectors and vector-borne pathogens

Genetics and molecular epidemiology of vector-borne diseases; Molecular biology and biology of host systems, vectors, pathogens and their interactions.

Ecology and wildlife management

Population`s genetics; Ecology and behavior at the fauna-livestock interface; Integrated monitoring of wildlife populations.

Animal Science

Biotechnology of reproduction.

IREC scientists will harmonize the collection of wildlife population data to improve health surveillance systems.

The applications of this research project in the field of health surveillance and disease control will improve our response to outbreaks of diseases shared between fauna, livestock and humans.

Scientists identify the best oral-bait delivery strategy for administering health treatments to wild populations of red deer

IREC scientists identify the most effective strategy for administering oral health treatments to wild populations of Iberian deer through the use of food baits

Skeletal anomalies in reared Senegalese sole (Solea senegalensis) juveniles: a radiographic approach.

Reared Senegalese sole Solea senegalensis Kaup show a high incidence of vertebral anomalies; however, little is known about its skeletal anomaly profile in the later farming phases. The purpose of this study was to provide a detailed description and quantification of the most common skeletal anomalies in reared Senegalese sole in the juvenile stage by means of computed radiography. A total of 374 Senegalese sole were classified according to the external morphology of the fish as normal or altered and then radiographed in latero-lateral and in dorso-ventral projections. Radiographic evaluation of anomalies focused especially on vertebral body anomalies (VBA) and vertebral column deviations (VCD). The 2 orthogonal projections provided a more complete visualization of the skeleton. Approximately 75% of the individuals showed at least 1 anomaly, while VBA and/or VCD were detected in 48.9% of the specimens. Regarding external morphology, 88% of the fish were categorized as normal, although about 72% of these normal fish displayed abnormalities in radiographies. The most frequent anomalies consisted of deformations of the caudal complex plates (hypurals, parhypural and epural), preurals and caudal vertebrae. Scoliosis was the most prevalent among VCD, affecting the caudal area in almost 15% of the individuals. The anomaly profile at the juvenile stages showed some differences compared to what has been reported previously in earlier stages of development. In light of these results, further investigation into the progression of skeletal anomalies over time and the causative factors at later stages is required.

Immunology, Virology

immune response of fish and shellfish

First description of a natural infection with spleen and kidney necrosis virus in zebrafish.

Zebrafish has become a popular research model in the last years, and several diseases affecting zebrafish research facilities have been reported. However, only one case of naturally occurring viral infections was described for this species. In 2015, infectious spleen and kidney necrosis virus (ISKNV ) was detected in zebrafish from a research facility in Spain. Affected fish showed lethargy, loss of appetite, abnormal swimming, distention of the coelomic cavity and, in the most severe cases, respiratory distress, pale gills and petechial haemorrhages at the base of fins. Cytomegaly was the most relevant histopathological finding in organs and tissues, sometimes associated to degenerative and necrotic changes. ISKNV belongs to the relatively newly defined genus Megalocytivirus , family Iridoviridae, comprising large, icosahedral cytoplasmic DNA viruses. This is the first case of naturally occurring Megalocytivirus infection in zebrafish research facilities, associated with morbidity. The virus has been identified based on both pathologic and genetic evidence, to better understand the pathogenesis of the infection in zebrafish and the phylogenetic relationship with other iridoviruses. Given the ability of megalocytiviruses to cross‐species boundaries, it seems necessary to implement stringent biosecurity practices as these infections may invalidate experimental data and have major impact on laboratory and cultured fish.

Susceptibility of Zebrafish to Vesicular Stomatitis Virus Infection.

The zebrafish, Danio rerio, has become recognized as a valuable model for infectious diseases. Here we evaluated the susceptibility of zebrafish to be infected with the mammalian vesicular stomatitis virus (VSV). Both zebrafish cells and embryos were highly susceptible to VSV infection. Mortalities exceeded 80% in infected embryos and were preceded by the invasion of the central nervous system by VSV. Live imaging of the infection with GFP-VSV as well as virus titration from infected fish confirmed the viral replication. Immunohistochemical analysis of embryonic fish provided evidence of viral antigens as well as of the apoptosis marker caspase-3 in the brain, eye, liver, pronephros, and skeletal muscle. So far, this is the first report describing the susceptibility of zebrafish to the mammalian virus VSV.

Long-term affected flat oyster (Ostrea edulis) haemocytes show differential gene expression profiles from naïve oysters in response to Bonamia ostreae

European flat oyster (Ostrea edulis) production has suffered a severe decline due to bonamiosis. The responsible parasite enters in oyster haemocytes, causing an acute inflammatory response frequently leading to death. We used an immune-enriched oligo-microarray to understand the haemocyte response to Bonamia ostreae by comparing expression profiles between naïve (NS) and long-term affected (AS) populations along a time series (1 d, 30 d, 90 d). AS showed a much higher response just after challenge, which might be indicative of selection for resistance. No regulated genes were detected at 30 d in both populations while a notable reactivation was observed at 90 d, suggesting parasite latency during infection. Genes related to extracellular matrix and protease inhibitors, up-regulated in AS, and those related to histones, down-regulated in NS, might play an important role along the infection. Twenty-four candidate genes related to resistance should be further validated for selection programs aimed to control bonamiosis.

Skeletal Anomalies in Senegalese Sole (Solea senegalensis), an Anosteocytic Boned Flatfish Species.

Skeletal anomalies affect animal welfare and cause important economic problems in aquaculture. Despite the high frequency of skeletal problems in reared Solea senegalensis, there is lack of information regarding the histological features of normal and deformed vertebrae in this flatfish. The aim of this study was to describe the histopathological and radiographical appearance of vertebral body anomalies. Sixty-seven juvenile fish were radiographically examined 104 or 105 days after hatching. Through radiographic images, vertebral segments were selected and processed for histopathological examination from 7 normal and 7 affected fish. Alterations in bone shape and vertebral fusion were the most significant anomalies in the vertebral bodies. These alterations occurred most frequently between the last 3 abdominal vertebrae and the first 10 caudal centra. Radiographically, deformed vertebrae showed flattening of the endplates and narrowing of the intervertebral spaces. The radiographic findings concurred with the histological lesions where affected vertebrae exhibited irregular endplates and changes in trabecular bone. Radiolucent cartilaginous tissue was evident in the endplates of the deformed vertebra and, in some cases, the cartilaginous material extended from the growth zone into the intervertebral space. These changes were likely the primary alterations that led to vertebral fusion. Fused vertebrae were often reshaped and showed a reorganization of the trabeculae. The formation of metaplastic cartilage is frequent in a variety of anomalies affecting teleost species.

Integrating genomic and morphological approaches in fish pathology research: The case of turbot (Scophthalmus maximus) enteromyxosis.

Enteromyxosis, caused by Enteromyxum scophthalmi, is one of the most devastating diseases stemming from myxozoan parasites in turbot (Scophthalmus maximus L.), being a limiting factor for its production. The disease develops as a cachectic syndrome, associated to catarrhal enteritis and leukocytic depletion, with morbidity and mortality rates usually reaching 100%. To date, no effective treatment exists and there are different unknown issues concerning its pathogenesis. The gross and microscopic lesions associated to enteromyxosis have been thoroughly described, and several morphopathological studies have been carried out to elucidate the mechanisms of this host-parasite interaction. More recently, efforts have been focused on a multidisciplinary approach, combining histopathology and transcriptome analysis, which has provided significant advances in the understanding of the pathogenesis of this parasitosis. RNA-Seq technology was applied at early and advanced stages of the disease on fishes histologically evaluated and classified based on their lesional degree. In the same way, the transcriptomic data were analyzed in relation to the morphopathological picture and the course of the disease. In this paper, a comprehensive review of turbot enteromyxosis is presented, starting from the disease description up to the most novel information extracted by an integrated approach on the infection mechanisms and host response. Further, we discuss ongoing strategies toward a full understanding of host-pathogen interaction and the identification of suitable biomarkers for early diagnosis and disease management strategies.

Early stages of Margaritifera margaritifera glochidiosis in Atlantic salmon: Morphopathological characterization.

Freshwater mussels of the order Unionida encyst into the fish mucosa to metamorphose and complete their life cycle, causing a parasitic disease known as glochidiosis. This parasitic stage represents a bottleneck for the survival of naiads, particularly for critically endangered species as Margaritifera margaritifera; however, little is known about the events occurring during this critical stage. Therefore, this study aimed to histologically characterize the development of M. margaritifera glochidiosis in Atlantic salmon to get insight into the pathogenesis of this interaction. Fish exposed to glochidia were sampled during the first 44 days post‐exposure, and organs were observed by stereomicroscopy and light microscopy. Glochidia attached to the gills by pinching the lamellar epithelium, whereupon an acute proliferative branchitis engulfed most of the larvae. However, during the first 14 days, a severe detachment of unviable glochidia occurred, associated with the presence of pleomorphic inflammatory infiltrate and epithelial degeneration. In the cases where larvae remained attached, a chronification of the lesions with none to scarce inflammation was observed. These results provide key information to better understand the complex host–parasite interaction during the early stages of glochidiosis and provide valuable information to optimize artificial rearing of naiads in conservation of threatened freshwater mussel populations.

Immunohistochemical expression of E–cadherin in different tissues of the teleost fish Scophthalmus maximus.

E–cadherin is an evolutionary conserved protein, whose main role as the principal component of adherens junctions is supporting epithelial cell–cell adhesion. It is an essential molecule for the maintenance of the epithelial barrier function and the analysis of its immunohistochemical expression is a valuable resource in morphopathological, ontogenetic and pathogenesis studies in mammals. As well, there is an increasing understanding of the importance of E-cadherin in the physiology of the immune system and the development of the immune response. Mucosal health is a primary issue in aquaculture research; nevertheless, there is a lack of immunohistochemical studies of cell junction proteins in fish species. In this work, an immunohistochemical technique was optimized in Bouin- and formalin-fixed paraffin-embedded tissues of turbot Scophthalmus maximus, employing a commercial antibody raised against human E-cadherin. The specificity of the antibody in recognizing the molecule in this teleost species was tested by western blot and mass spectrometry-based proteomic analyses. The assays showed a good specificity and indicated that the antibody recognizes the well conserved cytoplasmic domain of the protein. Immunohistochemistry showed the localisation of E-cadherin at cell-cell contact in the epithelia of the different organs, between the hepatocytes and the pancreatic acinar cells, as well as in the reticulo-epithelial stroma of the thymus. Also, the immunoreaction was observed in the cells constituting the melano-macrophage centres in the spleen and kidney. No immunostaining was detected, as expected, only in the heart and brain. No significant difference was noticed between the two fixative used for collecting the tissues samples. This is the first description of E-cadherin immunohistochemical expression in several tissues of a teleost. The immunohistochemical technique represents a useful tool to be used in the different areas of fish health research.

Effects of Enteromyxum spp. (Myxozoa) infection in the regulation of intestinal E-cadherin: turbot against gilthead sea bream

ABSTRACT: Enteromyxoses are relevant diseases for turbot and gilthead sea bream aquaculture. The myxozoan parasites invade the intestinal mucosa, causing a cachectic syndrome associated with intestinal barrier alteration; nonetheless, their pathological impact is different. Turbot infected by Enteromyxum scophthalmi develop more severe intestinal lesions, reaching mortality rates of 100%, whereas in E. leei ‐infected gilthead sea bream, the disease progresses slowly, and mortality rates are lower. The mechanisms underlying the different pathogenesis are still unclear. We studied the distribution and expression changes of E‐cadherin, a highly conserved protein of the adherens junctions, in the intestine of both species by immunohistochemistry and quantitative PCR, using the same immunohistochemical protocol and common primers. The regular immunostaining pattern observed in control fish turned into markedly irregular in parasitized turbot, showing an intense immunoreaction at the host–parasite interface. Nevertheless, E‐cadherin gene expression was not significantly modulated in this species. On the contrary, no evident changes in the protein distribution were noticed in gilthead sea bream, whereas a significant gene downregulation occurred in advanced infection. The results contribute to the understanding of the different host–parasite interactions in enteromyxoses. Host and parasite cells appear to establish diverse relationships in these species, which could underlie the different pathological picture.

Veterinary pathological diagnosis service

Provision of services to the public administration, animal production companies, veterinary professionals, zoos and aquariums for the pathological diagnosis of diseases in different animal species. • Provision of services and scientific collaboration with companies and research centers / groups for the evaluation of the effects of diets / treatments / vaccines in animal species, as well as for the optimization of production protocols. • Advice to companies and research centers / groups in the evaluation of the health status and well-being of production and experimental animals (team members have training certificates in the protection of experimental animals).

Development of experimental models of animal and human diseases in zebrafish

The experience gained in fish health and animal experimentation has led the group to develop a new line of research using zebrafish as a model for the study of diseases of interest in public health and veterinary and human medicine.

Assessment of skeleton genesis, skeletal and muscular abnormalities in different stages of production of farmed fish.

The objective of this line of research is the multidisciplinary approach to the main skeletal abnormalities, muscle growth and development in species of interest in aquaculture, mainly Senegalese sole, through the use of radiological, morphopathological and molecular techniques, with the purpose of develop tools aimed at the diagnosis and monitoring of skeletal abnormalities and growth under different production conditions.

Morphopathological study and local immune response in mucosa

The objective of this line of research is to address the knowledge of the intestinal, gill and skin mucosa, with the aim of improving health and developing protocols for evaluating the effect of diatas and treatments. To do this, we carried out a multidisciplinary approach using histopathological, immune and genetic techniques.

Pathogenesis and diagnosis of common and emerging diseases in mammals, birds and aquatic animals, with special emphasis on species of interest in aquaculture

Morphopathological and pathogenic characterization of common and emerging diseases, especially those that have an impact on animal production and the environment, using histopathological, immunohitochemical, molecular and electron microscopy techniques. The final purpose is to obtain new diagnostic tools and biomarkers that are of practical application to diagnosis, prevention, disease control and monitoring of animal health and welfare

Control of enteromyxosis of turbot and sparids. Characterization of the pathogenesis, the parasite-host interaction and the immune response.

Ex situ conservation of the endangered naiad Margaritifera margaritifera: Implementation of the cultivation action of the Life + Margal Ulla project (CAQ2016)

Consolidación GI_17072019-PG012 Anatomía Patológica Veterinaria

Search for tools for early diagnosis and detection of biomarkers of health / intestinal disease in turbot.

Melatonin for the first in vitro production of Iberian red deer embryos from dead animals.

Scientists from the IREC produce Iberian red deer embryos in vitro from germ cells of dead females for the first time, becoming the only ones in the world to produce embryos of this deer subspecies.

Mathematical models to improve the control of infectious diseases

A mathematical model allows understanding the epidemiological dynamics of the African swine fever (ASF), in which the wild boar is a key actor, constituting an essential tool to control epidemic outbreaks and prevent health emergency situations.

STOP the collection of Mediterranean spur-thighed tortoises as pets

The non-commercial collection of Mediterranean spur-thighed tortoises as pets in Morocco could compromise the conservation of wild populations in one of the hotspots of the species in northern Africa

Predation of tortoises by ravens: what is the role that plays the shrub?

Predation of hatchlings and juveniles of spur-thighed tortoises by common ravens could constitute a threat to the conservation of this chelonian in North African anthropic habitats with low shrub cover.

The social network of interactions between wild ungulates and livestock

The use of several complementary methodologies allows to identify in detail the social network of interactions between wild ungulates and livestock in areas where they coexist, providing the keys for the correct management of shared diseases by reducing the risk of pathogen transmission.

Antioxidants slow telomere shortening in white stork chicks

The relationship between oxidative stress and telomere shortening in vivo in free-living white stork chicks demonstrates the importance of physiological stress as a predictor of longevity and survival.

Good practices in relation to hunting wild boar and prevention of health risks

Nightjars wild boars are part of our natural environment and the management of their populations should be done sensibly and coexistence with other activities such as agriculture, animal husbandry, conservation and nature tourism. This manual of good practices It proposes a series of actions to help improve coexistence between wild boar and hunting use and other activities and resources of rural areas, with particular reference to control of their superabundance for the prevention of health risks.

Wild boar and African swine fever

Pollution owing to African swine fever (ASF) It is a highly contagious and lethal viral disease that affects both the domestic pig and the Wild boar. The virus has begun to spread throughout northern and eastern Europe, and if it reached Spain, the impacts on the pig sector, hunting and the environment would be dramatic.

The IREC participates in a program for the development of vaccines against ticks in Uganda

IREC scientists participate in a program for vaccine development dedicated to control ticks in cattle in Uganda. The project includes training Ugandan staff to begin production of the vaccine nationwide.

Vacuna frente a infestaciones provocadas por artrópodos hematófagos.

System for the expression of peptides on the bacterial surface

Antigenos recombinantes inmunoprotectores de anaplasma phacocytophilum, composición de las vacunas y sus usos.

Methods and compositions for tuberculosis diagnosis

Comederos para ciervos

Composición farmacéutica útil para la administración oral de fármacos y vacunas a jabalíes y sus aplicaciones.

Selective feeder for the administration of baits to wild boar piglets.

Vacuna frente a la tuberculosis

Production of embryos in deer and sheep by fertilization in vitro

MammalNet Project: Can we apply citizen science in monitoring mammals at a European level?

ENETWILD project: Harmonization in the collection and use of data on the distribution and abundance of wildlife in Europe

ADISRA Project: Wild birds as dispersants of antibiotic resistance

Proyecto E-RTA: Nuevas herramientas biotecnológicas para la vigilancia y el control de flavivirus emergentes en el reservorio aviar

CrimeaCaLM Project: Assessment of the current situation of the Crimean-Congo haemorrhagic fever virus in Castilla-La Mancha: Study on vectors and hosts

E-RTA Project: Analysis of new West Nile virus encephalitis outbreaks in Spain and their geographical expansion

WILD DRIVER Project: Effect of overabundance of deer and wild boar on emerging diseases

ANTICKUG Project: Development of a tick vaccine in Uganda

GALINFEC Project: Characterization of the immune response to the alpha-Gal antigen and its application to the control of infectious diseases

SUBINTERACT Project: The Subolesin / Akirine Interactome and its role in regulating the immune response in human and tick cells

Methods and compositions for tuberculosis diagnosis

Distribution patterns of saprolegniosis cutaneous lesions in wild and farmed brown trout (Salmo trutta L.) obtained using a geographic information system (GIS) (Fregeneda-Grandes y Aller-Gancedo , 2019)

A retrospective study was conducted using 250 clinical records of brown trout (Salmo trutta L.) with saprolegniosis by Saprolegnia parasitica, which had been collected from 8 rivers and 1 fish farm in the province of León (Spain). A geographic information system (GIS) was used to obtain skin lesion distribution patterns in males and females. Lesions in wild brown trout affected 15.31 ± 13.33% of the body surface, with a mean of 12.76 ± 6.56 lesions per fish. In addition, 51.23% of wild trout presented lesions with necrosis of the skin or fins. The pattern obtained when not distinguishing between sexes indicated that saprolegniosis lesions are mainly located above the lateral line and most frequently affect the dorsal cephalic region, the adipose fin, the peduncle and the caudal fin. However, differences were observed between males and females. Farmed trout presented a lower percentage of affected body surface (2.06 ± 4.36) and a lower number of lesions with and without necrosis because they received preventive treatment for saprolegniosis.

First record of an outbreak of saprolegniosis by Saprolegnia parasitica in Pseudochondrostoma duriense (Coelho, 1985) (Cyprinidae)(Aller-Gancedo et al., 2016)

An outbreak of saprolegniosis by Saprolegnia parasitica in the cyprinid fish Pseudochondrostoma duriense (Coelho, 1985) is described here for the first time. P. duriense is endemic to the Iberian Peninsula. Hundreds of sick and dead fish appeared in the River Bernesga (Spain). Skin and subcutaneous skeletal musculature were infected. Fish showed multi-focal dermatitis with loss of the epidermis, degeneration and necrosis of the skeletal muscle fibres, ulcerative keratoconjunctivitis and anterior uvitis.

Biocontrol of saprolegniosis in rainbow trout (Oncorhynchus mykiss Walbaum) using two bacterial isolates (LE89 and LE141) of Pseudomonas fluorescens (González-Palacios et al., 2019)

The probiotic activity of 15 bacterial isolates that inhibit Saprolegnia parasitica in vitro was tested for the biocontrol of saprolegniosis in rainbow trout (Oncorhynchus mykiss Walbaum), adding the bacteria to tank water for 14 days at a concentration of 106 bacteria ml−1 water. Pseudomonas fluorescens LE89 and Pseudomonas fluorescens LE141 were effective in controlling experimental infection with S. parasitica since of the fish treated with LE89, 24.5% ± 16.27% (p < 0.05) became infected, as did 42.8% ± 8.41% (p < 0.05) of those treated with LE141. Given their protective effect when administered in water, their effect was also studied when administered in feed before and after experimental infection. Both bacterial isolates survived low pH levels and the action of bile, grew in skin and intestinal mucus, were resistant to several antibiotics and survived in feed; however, neither of the two isolates prevented S. parasitica infection when administered in feed.

Exosomes and their use as vaccine

Jordi Figuerola publicactions

Resumen de publicaciones en Google Academico

Pig as a experimental model

the group is interested in using large animals for cellular therapies. Dendritic cells (DCs) are professional antigen presenting cells that play a crucial role in both initiation and modulation of the immune response. Due to their physiological properties and the availability of clinical grade reagents, immunostimulatory DCs have been safely and successfully used in clinical trials designed to stimulate immune responses against tumours and pathogens. Our work aims to address fundamental questions in DC therapies by using immunologically relevant animal models, more informative than mice. Domestic pigs share many similarities with humans and represent an excellent animal model for immunological studies. We have previously set up a model in pigs for DC tracking in vivo using magnetic resonance imaging that we will use in these experiments.

Reverse vaccinology

A major drawback in vaccine development in animal health is the fact that antigenic peptides presented by MHC class I (in pigs SLA-I) have not been fully characterized, hampering the rational design of vaccines. Identification and characterization of B- and T-cell epitopes (also known as antigens) for a given pathogen has proved crucial for understanding the basic mechanisms of immunological protection and for the rational design of effective vaccines. Protective immunity is usually mediated by B lymphocytes, CD8+/CD4+ T lymphocytes or both. Recognition of epitopes by lymphocytes from different species and individuals is restricted by the major histocompatibility complex (MHC) molecules – named swine leukocyte antigen (SLA) in pigs- which is responsible for foreign antigen presentation. In the case of viral infections, classical SLA class I (SLA-I) molecules act as molecular cradles that present endogenous and foreign peptides or epitopes to cytotoxic CD8+T cells (CTL) at the cell surface of healthy and infected cells. Thus, they provide the immune system with a mechanism for interrogating the viral proteome from the outside of infected cells. Our team and in collaboration with other international groups have applied reverse vaccinology to identify cross-reacting MHC class I T-cell epitopes from two different Swine Influenza virus (SwIV) H1N1 lineages in pigs presented by SLA-1*0702.

Afrcan swine fever immunology

ASFV causes severe disease of domestic pigs that can result in up to 100% mortality with concomitant serious socio-economic impacts. Nowadays, there is no vaccine against ASFV. Historically, vaccine development has been brought about by an essentially empirical approach where antibodies were usually the major protective factor. However, there are numerous pathogens for which this approach has not proved protective, including ASFV, for which the reasons remain unclear. Thus, other vaccination approaches are required, which can only be informed by a deeper knowledge of immune responses during host-pathogen interactions. The work in the group is directed into a better understanding of the immune responses required for protection in order to provide the information for rationally design a vaccine against ASFV or SwIV.

Pig Influenza

In the last decades, the mouse model tremendously contributed to the progress of research in immunology. However, it is worth noticing that research in large/unconventional animals has given pivotal information to immunology in general. The need for translational immunological research, the development of new animal models more adapted to the scientific question, and the acquisition of new knowledge in comparative immunology is increasing in importance every year. The main aim of the group is to provide insights into immunological host-pathogen interactions by studying the immune system of the pigs in the context of natural relevant viral infections, such as Influenza virus (IV) or African swine fever virus (ASFV). Also, pigs will be used as model for celular therapies. Besides its importance as a zoonotic agent, Swine Influenza Virus (SwIV) is also important for its impact in the pig farming industry. Swine influenza is a relevant porcine respiratory disease, whose importance has been often underestimated due to the impact of other respiratory pathologies. Our data in collaboration with HIPRA S.A. have shown by phylogenetic analysis that circulating SIV strains in Spain belonged to the lineages “Avian-Like” H1N1, “Human-Like” H3N2, and “Human-Like” H1N2, showing tight relationships with early or contemporary strains described in Europe. Also, our results indicated that reassortment events could have influenced the evolution, indicating that circulating SIV strains in pig farms could be a potential zoonotic agent. Thus, control of SwIV is very important not only to prevent pulmonary lesions in pigs but also to control a zoonotic infection.

MOST FREQUENT INFECTIONS, ANTIBIOTIC CONSUMPTION AND PRESENCE OF RESISTANCE

MOST FREQUENT INFECTIONS, ANTIBIOTIC CONSUMPTION AND PRESENCE OF RESISTANCE

EFFICACY OF PHYTOPHARMACS IN ANIMAL PRODUCTION

EFFICACY OF PHYTOPHARMACS IN ANIMAL PRODUCTION

USE OF NUTRACEUTICALS AS AN ALTERNATIVE TO ANTIBIOTICS AND EVALUATION OF THEIR FUNCTIONALITY IN ANIMAL PRODUCTION

USE OF NUTRACEUTICALS AS AN ALTERNATIVE TO ANTIBIOTICS AND EVALUATION OF THEIR FUNCTIONALITY IN ANIMAL PRODUCTION

DRUG RESIDUES IN DIFFERENT FLUIDS AND ANIMAL TISSUES

Determination and quantification of drug residues in different fluids and tissues

PHARMACOKINETICS AND PHARMACODYNAMICS OF ANTIMICROBIALS IN VETERINARY MEDICINE

Pharmacokinetic study of different antimicrobials such as fluoroquinolones, macrolides, beta-lactams in different animal species such as rabbit, sheep, goat, horse, sea turtle ... Susceptibility tests, cut point calculation, PK / PD integration

Ecology and evolution of zoonoses of relevance in public health

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Borrelia burgdorferi sensu lato prevalence and diversity in ticks and small mammals in a Lyme borreliosis endemic Nature Reserve in North-Western Spain. Incidence in surrounding human populations.

To determine the prevalence and diversity of Borrelia burgdorferi sensu lato (s.l.) in an endemic Nature Reserve (Sierra del Sueve) in North-Western Spain, and the risk of human exposure to infected ticks in Asturias, 1013 questing ticks and 70 small mammals were collected between 2012 and 2014. A retrospective descriptive analysis was also carried out on human Lyme borreliosis (LB) cases reported to the local hospital (Cabueñes). Samples were screened for B. burgdorferi s.l. presence by a nested PCR assay, and genospecies were confirmed by sequencing. B. burgdorferi s.l. was detected in 1.4% (12/845) of I. ricinus questing nymphs, 9.1% (2/33) of questing adults, and 12.9% (9/70) of small mammals, as well as in the other tick species. PCR positive samples of 17 questing tick and 6 small mammals were sequenced. Four genospecies were identified: B. afzelii, B. garinii, B. lusitaniae, and B. valaisiana. Phylogenetic analyses based on the flaB gene showed the heterogeneity of B. afzelii in this area. The detection of B. burgdorferi s.l. among questing ticks and small mammals in the study area, as well as the abundance of ticks and of large wild and domestic mammals, indicate a high risk of infection by B. burgdorferi s.l. in the area. Reporting of LB cases to the local hospital support this, and shows the need of thorough monitoring of B. burgdorferi infection in ticks and hosts in the area. More investigations are needed to assess the role of different wildlife species and the risk of transmission to humans.

Vector-borne diseases

Epidemiology of tick-borne diseases of interest in animal (piroplasmosis and anaplasmosis) and human health (Lyme borreliosis, rickettsiosis and Q fever) Molecular genetics of tick-borne pathogens

Louping ill in goats, Spain, 2011.

Although louping ill affects mainly sheep, a 2011 outbreak in northern Spain occurred among goats. Histopathologic lesions and molecular genetics identified a new strain of louping ill virus, 94% identical to the strain from Britain.

A one-step TaqMan real-time qRT-PCR assay for the specific detection and quantitation of the Spanish goat encephalitis virus (SGEV).

Louping ill-like virus (LI) has been recently detected in two different locations in the north of Spain and separated by only around 400 km. Using molecular approaches, the viruses causing both outbreaks have been shown to be different to LI virus, but also different to each other. They have been called SSEV (Spanish sheep encephalitis virus) and SGEV (Spanish goat encephalitis virus) taking into account the species from which they were isolated. The aim of this paper was to design a quantitative TaqMan real-time RT-PCR protocol, for the specific diagnostic and quantitation of SGEV. Linearity, efficiency and dynamic range as well as reproducibility and specificity of the method has been tested and established.

First Confirmation of Schmallenberg Virus in Cattle in Spain: Tissue Distribution and Pathology.

Between January and June 2013, nine stillborn bovine foetuses with congenital malformations from nine cattle herds located in Salamanca (central Spain) were detected. Necropsy was performed on two calves. Pathological lesions together with molecular genetics and serological results allowed a definitive diagnosis: first confirmation of Schmallenberg virus (SBV) infection in cattle in Spain. SBV was detected in different tissues and organic fluids in both animals including blood, suggesting a possible viraemia. The umbilical cord was also positive for the presence of SBV in both animals. The former tissue provides an easy to obtain sample and might be a sample of choice when necropsy is carried out in the field.

Canine adenovirus type 1 (CAdV-1) in free-ranging European brown bear (Ursus arctos arctos): A threat for Cantabrian population?

Canine adenovirus type 1 (CAdV-1) is responsible for infectious canine hepatitis. The disease has been described in captive American black bear (Ursus americanus) and European brown bear (Ursus arctos arctos), with just one recently reported case in a cub of a free-ranging brown bear (Ursus arctos horribilis) from Alaska. The aim of this work is to summarize findings related to presence and associated mortality of CAdV-1 in 21 free-ranging Cantabrian brown bears (Ursus arctos arctos) submitted to necropsy in Asturias and Castilla y León (northwestern Spain) from 1998 to 2018.

Ictiopathology

Laboratory diagnosis and support in the eradication of the main salmonid diseases.

Mycobacteria

Study of tuberculosis in species that can contribute to the maintenance of bovine tuberculosis in Asturias. Especially in the badgers and wild boars. Bovine paratuberculosis. Isolation and typing of strains in both domestic and wild ruminants. Molecular epidemiology and diagnostic tests.

Tuberculosis Epidemiology and Badger (Meles meles) Spatial Ecology in a Hot-Spot Area in Atlantic Spain (Acevedo et al. 2019)

We provide a temporal overview (from 2012 to 2018) of the outcomes of tuberculosis (TB) in the cattle and badger populations in a hot-spot in Asturias (Atlantic Spain). We also study the badger`s spatial ecology from an epidemiological perspective in order to describe hazardous behavior in relation to TB transmission between cattle and badgers. Culture and single intradermal tuberculin test (SITT) were available for cattle as part of the National Program for the Eradication of TB. A field survey was also carried out in order to determine the paddocks and buildings used by each farm, and the information obtained was stored by using geographic information systems. Moreover, eighty-three badgers were submitted for necropsy and subsequent bacteriological studies. Ten badgers were also tracked, using global positioning system (GPS) collars. The prevalence of TB in cattle herds in the hot-spot increased from 2.2% in 2012 to 20% in 2016; it then declined to 0.0% in 2018. In contrast, the TB prevalence in badgers increased notably (from 5.55% in 2012-2015 to 10.64% in 2016-2018). Both cattle and badgers shared the same strain of Mycobacterium bovis. The collared badgers preferred paddocks used by TB-positive herds in spring and summer (when they were more active). The males occupied larger home ranges than the females (Khr95: males 149.78 ± 25.84 ha and females 73.37 ± 22.91 ha; Kcr50: males 29.83 ± 5.69 ha and females 13.59 ± 5.00 ha), and the home ranges were smaller in autumn and winter than in summer. The averages of the index of daily and maximum distances traveled by badgers were 1.88 ± (SD) 1.20 km and 1.99 ± 0.71 km, respectively. One of them presented a dispersive behavior with a maximum range of 18.3 km. The most preferred habitat was apple orchards in all seasons, with the exception of winter, in which they preferred pastures.

Protective Effect of Oral BCG and Inactivated Mycobacterium bovis Vaccines in European Badgers (Meles meles) Experimentally Infected With M. bovis (Balseiro et al. 2020)

In Europe, badgers (Meles meles) are recognized as major tuberculosis (TB) reservoir hosts with the potential to transmit infection to associated cattle herds. Recent studies in Spain have demonstrated that vaccination with a heat-inactivated Mycobacterium bovis vaccine (HIMB) successfully protects captive wild boar and red deer against progressive disease. The aim of this study was to evaluate the efficacy of two oral vaccines against TB in a badger model: the live-attenuated M. bovis bacillus Calmette-Guérin BCG vaccine (Danish strain) and a HIMB vaccine. Twenty-four badgers were separated in three treatment groups: oral vaccinated with live BCG (108 CFU, n = 5), oral vaccinated with HIMB (107 CFU, n = 7), and unvaccinated controls (n = 12). All badgers were experimentally infected with M. bovis (103 CFU) by the endobronchial route targeting the right middle lung lobe. Throughout the study, clinical, immunological, pathological, and bacteriological parameters of infection were measured. Both vaccines conferred protection against experimental TB in badger, as measured by a reduction of the severity and lesion volumes. Based on these data, HIMB vaccination appears to be a promising TB oral vaccine candidate for badgers in endemic countries.

Pilot project to improve biosecurity in extensive cattle in Valle de Alcudia

Integrated strategies for tuberculosis control and eradication in Spain (ERATUB)

WILDLIFE HEALTH

Tuberculosis (badger and wild boar): Biosecurity / Vaccine efficacy in badgers / epidemiology / development of diagnostic techniques Amphibian diseases: Ranavirus (epidemiology / diagnostic techniques) Vector-borne diseases: Flavivirus (epidemiology and vaccine efficacy tests)

HEALTH AND PATHOLOGY OF RUMINANTS

Infectious and parasitic diseases of ruminants (paratuberculosis, toxoplasma) Sporadic and reproductive pathology Pathogenesis and immunopathology of the main diseases

RESPIRATORY PATHOGENS OF BACTERIAL ETIOLOGY

Porcine Respiratory Complex: virulence factors, diagnosis, alternatives to the use of antibiotics for treatment and vaccines (Streptococcus suis, Actinobacillus pleuropneumoniae ...) Epidemiological studies of virulence factors and antimicrobial resistance in Francisella tularensis isolated from human and animals in Castilla y León

CONTINENTAL AQUACULTURE AND ICTYOPATHOLOGY

Continental aquaculture Infectious fish diseases Immunoprotection against fish diseases Saprolegnia infections Cell cultures and in vitro assays in aquaculture

DIGESTIVE DISEASES OF SWINE

Enteric infectious processes in pigs Modulation and improvement of digestive microbiotics. Prebiotics and probiotics Monitoring and reduction strategies for the development of resistance to antibiotics in pigs. Alternatives to the use of antibiotics in animal production

TROPICAL AND PARASITIC DISEASES

CONTROL OF DISEASES PRODUCED BY HELMINTH PARASITES IN RUMINANTS Detection of antihelmintic resistance Search for new molecules with antihelmintic activity Development of new ´point of care´ diagnostic methods for on-farm detection of gastrointestinal nematode infections Search for genetic markers related to resistance to infection by gastrointestinal nematodes

Emerging and Transboundary Diseases

Team Manager: Miguel Angel Jiménez-Clavero majimenez@inia.es International Reference Laboratories UE and FAO for African Swine Fever: Technical Director EURL and WRC FAO for ASF : Maria Luisa Arias Neira arias@inia.es

Biological Alert Laboratory Network (RE-LAB)

INIA-CISA takes part of the Biological Alert Laboratory Network (RE-LAB), under the Presidency of the Government and dedicated to dealing with threats caused by biological agents. Coordinators INIA-CISA for RELAB: Miguel Angel Jiménez (majimenez@inia.es) y Alejandro Brun (brun@inia,es)

Network of High Biosafety Laboratories (RLASB)

The INIA-CISA, with IRTA-CReSA, belongs to the Network of High Biosafety Laboratories (RLASB) and offers the possibility to work in its Biosafety facility. Coordinators INIA-CISA for RELASB: Dirección ( Maria Luisa Arias Neira arias@inia.es) y Dirección técnica, Lara del Rio delrio.lara@inia.es; INIA-CISA is Reference Centre in Biosafety for FAO (INIA-CISA) Technical Director: Gonzalo Pascual Álvarez gpascual@inia.es;

Molecular and Cellular Biology of Prions

Team Manager: Juna María Torres Trillo jmtorres@inia.es

Fish Immunology and Pathology

Team Manager: Carolina Tafalla Piñeiro tafalla@inia.es

New Strategies for the Control of relevant Pathogens in Animal Health

Team Manager: Noemí Sevilla Hidalgo sevilla@inia.es

Immunoprofilaxis of Vector-Borne Viral Diseases

Team Manager: Alejandro Brun Torres brun@inia.es

Epidemiology And Environmental Health

Team Manager: Ana de la Torre Reoyo torre@inia.es

Cholangiocarcinoma in a Free-Ranging Eurasian Brown Bear (Ursus arctos arctos) from Northern Spain.

A hepatic cholangiocarcinoma with metastases in the gallbladder, left elbow joint, adrenal glands, and lungs was observed in a female 21-yr-old free-ranging Eurasian brown bear (Ursus arctos arctos) found in the Principality of Asturias (northern Spain). Gross and histopathologic findings are described.

RNA-Seq analysis of ileocecal valve and peripheral blood from Holstein cattle infected with Mycobacterium avium subsp. paratuberculosis revealed dysregulation of the CXCL8/IL8 signaling pathway.

Paratuberculosis is chronic granulomatous enteritis of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Whole RNA-sequencing (RNA-Seq) is a promising source of novel biomarkers for early MAP infection and disease progression in cattle. Since the blood transcriptome is widely used as a source of biomarkers, we analyzed whether it recapitulates, at least in part, the transcriptome of the ileocecal valve (ICV), the primary site of MAP colonization.

RTI2018-096010-B-C21- Badger (Meles meles) and animal tuberculosis in Spain: badger-bovine interaction in hospot areas and disease control measures at the interface

RTA2017-00055-C02- INIA- Q fever research in Asturias. Study of the domestic and wild cycle of Coxiella burnetii infection and its relationship with human cases

RTI2018-094192-R-C22- Development of new diagnostic methods based on bovine biomarkers (proteins) for early diagnosis of bovine paratuberculosis

Study of the main pathogens of eels (Anguilla anguilla) in wild populations of the Asturias´ rivers

Immunotherapies

Basic and traslational research focus in new immunotherapies for pets.

Development of vaccines against the Porcine Reproductive and Respiratory Syndrome (PRRS)

Development of vaccines against the Porcine Reproductive and Respiratory Syndrome (PRRS)

Peripartum diseases in cattle: description of biomarkers and assessment of their economic impact

Peripartum diseases in cattle: description of biomarkers and assessment of their economic impact

Food-borne zoonoses

Food-borne zoonoses

Epidemiology and diagnosis of infections caused by protozoan parasites

Epidemiology and diagnosis of infections caused by protozoan parasites

Study of the virulence factors, pathogenesis mechanisms, and biological variability of protozoan parasites

Study of the virulence factors, pathogenesis mechanisms, and biological variability of protozoan parasites

Development of control tools (drugs and vaccines) for protozoan diseases in ruminants

Development of control tools (drugs and vaccines) for protozoan diseases in ruminants

Development of in vitro, ex vivo and in vivo experimental models for protozoan diseases in ruminants

Development of in vitro, ex vivo and in vivo experimental models for protozoan diseases in ruminants

OIE Reference Laboratory for African swine fever

World Organisation for Animal Health (OIE) Reference Laboratory for African swine fever

OIE Reference Laboratory for African horse sickness

World Organisation for Animal Health (OIE) Reference Laboratory for African horse sickness

European Union Reference Laboratory for Bovine Tuberculosis

The VISAVET Health Surveillance Centre (Universidad Complutense de Madrid, Spain) was appointed as the European Union Reference Laboratory for Bovine Tuberculosis in 2008 (Commission Regulation (EC) No 737/2008)

Minimal Invasion Surgery, Cell Therapy, Bioengineering, Assisted Reproduction, Cardiovascular

The main strategic lines of research that make up the scientific program of this Group are structured in 5 areas: Minimal Invasive Surgery (A), Cell Therapy (B), Bioengineering (C), Assisted Reproduction (D) and Cardiovascular (E), coordinated by Dr. Francisco Miguel Sánchez Margallo. A. Area of Minimally Invasive Surgery A. 1. Laparoscopy. A. 2. Endoscopy A. 3. Microsurgery A. 4. Anesthesiology A. 5. Endoluminal Diagnostics and Therapeutics B. Cell Therapy Area C. Bioengineering Area D. Assisted Reproduction Area E. Cardiovascular Area

Facilities

Design, production and characterization of bio- and nanomaterials, body tissues, medical devices and systems, from a physico-chemical, functional, toxicological and biological perspective as part of NANBIOSIS, one of the Spain’s 29 Unique Scientific and Technological Infrastructures (ICTS) recognised by the Consejo de Política Científica, Tecnológica e Innovación (CPCTI of the MINECO) in collaboration with the Centre for Biomedical Research Network in Biomaterials, Bioengineering and Nanomedicine (CIBER-BBN). - U14. Cell Therapy. http://www.nanbiosis.es/portfolio/u14-cell-therapy/ - U19. Clinical Tests Lab. http://www.nanbiosis.es/portfolio/u19-clinical-tests-lab/ - U21. Experimental Operating Rooms. http://www.nanbiosis.es/portfolio/u21-experimental-operating-rooms/ - U22. Animal Housing. http://www.nanbiosis.es/portfolio/u22-animal-housing/ - U23. Assisted Reproduction. https://www.nanbiosis.es/portfolio/u23-assisted-reproduction/ - U24. Medical Imaging. https://www.nanbiosis.es/portfolio/u24-medical-imaging/

Clinical development of a cell therapy for cardiac repair based on tissue engineering

Prostatic Apoptosis induced by Prostatic Artery Occlusion (PAO): Study of proof of concept in the pathological process and potential clinical applications

A pilot line for the next generation of smart catheters and implants

Escherichia coli.

An active line is the study of verotoxigenic and enterohemorrhagic E. coli, analyzing the epidemiological importance of wild ungulates as a natural reservoir and their interrelation with infection in sheep and cattle, and in the human species, by phenotypic and genotypic typing.

Antibiotic resistance

Characterization of antibiotic resistance in clinical isolates of medical and veterinary interest, and the search for alternatives to the use of antibiotics in the field of animal health and medicine.

Tuberculosis

For more than 25 years, we have been working in the tuberculosis research line, both in domestic animals and in wild artiodactyls, as well as in humans, contributing to characterize the epidemiology of animal tuberculosis, to improve the control of wild tuberculosis and to eradication of domestic tuberculosis.

Leishmaniosis

This line focuses on the study of the disease in dogs, especially in the development of vaccines against canine Leishmaniosis and new diagnostic techniques.

Ticks

Study of the distribution, speciation and characterization of ticks at different geographical levels. Tick-borne diseases, especially piroplasmosis and some arbovirosis, are also studied.

Mosquitoes and sandflies as disease vectors.

Surveillance studies of Aedes albopictus and flavivirus transmitting vectors such as Usutu or West Nile virus. Epidemiological studies are being conducted among hosts susceptible to these flaviviruses, especially among mosquitoes, birds, horses and humans. As for sandflies, studies of distribution and speciation and analysis of phlevovirus (Tuscany, Sicily, etc.) are carried out on insects and other hosts (animals and humans).

Network of High Biosafety Level Laboratories (RLASB) - ICTS (Spanish Scientific and Technical Singular Infraestructures)

The High Biological Safety Laboratories Network (RLASB) is a Unique Scientific and Technical Infrastructure (ICTS) of High Biological Safety laboratories that offers the possibility of using integrated laboratory and animal services, exclusively dedicated to work with infectious agents that demand conditions of high biological security for its manipulation. Due to its size, size and capacity, both High Security Biological Installations are unique at a national and international level, given that there are very few laboratories of these characteristics and this capacity. The RLASB is a distributed ICTS infrastructure formed by two nodes.: (1) BCL-3 of the Animal Health Research Center INIA-CISA, and (2) BCL-3 of the Animal Health Research Center IRTA-CReSA.

Exotic diseases subprogram

- Pathogenesis and prophylaxis of Classical Swine Fever (CSF) and other Pestivirus infections - Pathogenesis and prophylaxis of African Swine Fever (ASF) - Arboviruses and vectors - Pathogenesis, diagnosis and control of avian viral infections - Pathogenesis and prophylaxis of Coronavirus infections - Prion diseases

Exotic diseases subprogram

- Immunopathogenesis and protection against PRRSV (Porcine reproductive and respiratory syndrome virus) - Pathogenesis, epidemiology and control of infections caused by ssDNA viruses - Viral Evolution - Respiratory bacterial infections - Zoonotic bacterial infections and antimicrobial resistance with one health perspective - Tuberculosis - Microbiota, gut health, and alternatives to antibiotics

Epidemiology & risk assessment subprogram

- Descriptive and analytical epidemiology - Risk Assessment - Disease transmission modelling - Sociology in Animal Health - Biosecurity at farm level

Biosafety level 3 Area (BSL3)

Facility that has BSL-3 biosafety laboratories and BSL-3 necropsy room working with biological agents listed in Risk Group 3 (former OIE List A). This area has also a ABSL-3 animal facilities with three individual boxes with air conditioning and independent access to allow the simultaneous performance of different assays and controls.

SEVISEQ Equine Health Surveillance Unit

SEVISEQ (Equine Health Surveillance Unit) is created due to the need of support for the official institutions and is focused on several aspects: the high presence of equine infectious diseases in Spain, the existence of clinical outbreaks, the lack of awareness regarding the importance of equine infectious diseases by horse owners and professionals of the equine industry, and the importance of a correct application of prevention and control measures in every equine premises. The work at SEVISEQ is divided into six lines: - Diagnosis of acute infection. - Analysis of samples prior to exportation and sales. - Analysis of samples for the annual health surveillance in breeding stud farms. - Technical assistance to veterinary surgeons. - Production of health surveillance reports. - Research in equine infectious diseases.

Mycobacteria Unit

Mycobacteria Unit routinely performs bacteriological and molecular diagnose of infections produced mainly by members of the Mycobacterium tuberculosis and avium complexes. Moreover, it realizes studies of molecular and epidemiological characterization. The main lines of research are to set up new technologies for the diagnosis (in vivo e in vitro) of the main principal diseases caused by mycobacteria, as well as the development of new technology for molecular characterization studies. The laboratory also designs field studies of control and eradication programs in problem situations for subsequent implementation at a national level. The laboratory also advises Animal Health Services of the Autonomous Communities, farmers and veterinarians, as well as Ministries. In addition as European Union Reference Laboratory for Bovine Tuberculosis, they provide technical and scientific support to all National Reference Laboratories (NRL) and the official veterinary services of the European Union.

Viral Immunology and Preventive Medicine Unit

Viral Immunology and Preventive Medicine Unit has been developed several lines of research related to the diagnosis of animal viruses (African swine fever, classical swine fever, African horse sickness, bluetongue…) And risk analysis of possible entry and spread of infectious diseases in our country (in addition to those mentioned above, also foot & mouth disease, West Nile, avian flu) The group also develops the tasks of the reference laboratory for the OIE to African swine fever and African horse sickness, and has extensive experience in the management of these and other diseases such as classical swine fever. He also actively collaborates with OIE, FAO, OIRSA, DG SANCO and EFSA. The scientific interest of the service focuses on the study of infectious diseases in animals and in particular the development of new diagnostic techniques, new generation vaccines and new strategies for their control through risk analysis and modeling, as well as establishment of networks for health monitoring.

Foodborne Zoonoses and Antimicrobial Resistance Unit

Foodborne Zoonoses and Antimicrobial Resistance Unit became operational in 1996 based on the Veterinary Antimicrobial Resistance Surveillance Network (VAV). Nowadays, it includes the study of different food-borne zoonoses (Salmonella, Campylobacter, E. coli, Yersinia, Staphylococcus aureus ...). It is also responsible for analyzing antimicrobial resistance in zoonotic organisms, commensal bacteria of the intestinal tract or clinical isolates. The group has several lines of research on the characterization of important foodborne pathogens for Public Health and the mechanisms involved in the emergence and spread of antimicrobial resistance by using molecular tecniques as well as sequencing. It is outstanding the leadership of this group in international research projects and its collaboration with Public Administrations and companies. ZTA gives support to the Ministry of Agriculture, Fisheries, Food and Environment (MAPAMA) and the Community of Madrid developing surveillance programs for zoonotic agents.

Pathology and Forensic Veterinary Medicine Unit

Pathology and forensic veterinary medicine unit currently works on routine diagnosis of infectious and nutritional outbreaks that originate in animal production, mainly pigs, small ruminants and aquaculture. We actively participate in the description of the pathological mechanisms that occur in the development of new vaccines and drug treatments , both experimentally and farms, identifying potential adverse effects. Our group has wide experience in the field of experimental cancer models both the histopathological and immunohistochemical studies, as well as the application of new diagnostic tools such as PROFILING AND IMAGING (MALDI TOF). Actually, our major effort is focused on the development and application of these new technologies to the field of pathology image.

Neglected and Emerging Diseases Unit

Servicio de Zoonosis Emergentes, de Baja Prevalencia y Agresivos Biológicos realiza tareas de vigilancia epidemiológica y monitorización de patógenos emergentes y/o ignorados en distintas especies hospedadores (y en los posibles vectores en su caso). Algunos de estos agentes infecciosos presentan una aparente baja prevalencia, lo que hace que no sean tenidos en cuenta como prioridades en la mayoría de programas de Sanidad Animal a pesar de las graves consecuencias que puede implicar su presencia. La realidad es que la situación real de muchas de estas enfermedades se desconoce, y debido a los cambios en el clima y al incremento en los movimientos migratorios se han creado situaciones favorables para su propagación en entornos inicialmente libres de las mismas. La vigilancia epidemiológica constituye una herramienta útil y eficaz en la prevención y control de posibles brotes. El servicio colabora con administraciones locales y de comunidades autónomas en la monitorización de varias enfermedades.

Microbial Identification and Characterisation Unit

This unit is focused on the study at the phenotypic, genomic and protein leves of pathogenic bacteria of veterinary interest. The main activities of the Unit are divided into three research lines that are closely related: clinical diagnosis (using phenotypic, molecular and chemotaxonomic methods, the molecular characterization of pathogenic bacteria of relevant clinical significance for epidemiological purposes and description of new bacterial species (bacterial taxonomy). The molecular characterization allows tracing epidemiological links between strains associated with outbreaks of disease, global epidemiological studies, monitoring programs, etc. Studies of bacterial taxonomy not only allow the description of new species but also allow, for example, the association of different pathogens with atypical clinical processes.

Bacterial taxonomy

Traditional systems of bacteriological diagnosis are sometimes not sufficient to achieve a definitive identification of the etiological agent. As a result, certain pathogens may be misidentified or unidentified. In these circumstances, it is advisable the use of alternatives to traditional techniques. One of the methodologies used by our group is the sequencing of the 16S rRNA gene. The application of this methodology, together to other phenotypic, quimiotaxonomic and genetic methods has led to the description by our group of new species and bacterial genera isolated from different animal species and associated, in some cases, to different clinical process (Corynebacterium mastitidis, Corynebacterium suicordis, Uruburuella suis, Pseudomonas simiae, Streptococcus entericus as a few examples). - Association of old pathogens to new clinical process and/or new animal hosts. - Descripction of new bacterial species

Molecular typing

The epidemiological study of infectious diseases is used to determine the clonal relationship between various isolates of the same bacterial species. Different techniques can be used to distinguish and classify strains of a given bacterial species. This discrimination at the strain level is useful for understanding different aspects of the epidemiology of bacterial diseases. Our group has long been using various techniques such as pulsed field electrophoresis or multicolcus sequence typing for the molecular characterization of a broad spectrum of bacterial pathogens in different livestock species. The application of these techniques has allowed to obtain results, in some cases, of great epidemiological value. - Resarch of existence and extent of an outbreak - Identification of reservoirs and sources of infection - Monitoring the dissemination of particular clones, etc..

Clinical diagnosis

One task that has always demanded great attention by our group, both from the point of views of teaching and research is that related with animal diseases. This line of work has been a huge boost from the beginning, doing continued efforts to deepen the knowledge of the different pathogens involved in outbreaks of disease. The group has a great background in ovine mastitis, in the diagnostic of infectious diseases of wild and exotic animals, as well as bacterial diseases in pigs and fish. - Development of new techniques for more efficient diagnosis - Advance in the knowledge of the transmission mechanisms and survival to technological treatments.

Application of MALDI-TOF technique in the clinical microbiology and Agri-food field

MALDI-TOF technique has been traditionally used for proteomics studies, mainly, for the identification and characterization of proteins of interest. Recently, this analitycal technique has been demonstrated to be useful in the microbiology field not only for identification but also for characterization of microorganisms, mainly bacteria and fungi. - Microbial identification based on the MALDI-TOF mass profile. - Typing and subtyping studies susing MALDI-TOF approach. - Protemic assays: usefulness in Veterinary/Food Safety

Characterization of the emergence of foodborne pathogens subjected to surveillance and monitoring programs through quantitative epidemiology

Development and application of quantitative tools to the study the epidemiology of important diseases in public and animal health, including the evaluation of control and eradication programs of animal diseases, the spatial analysis, the molecular epidemiology and the identification of risk factors related to human and animal pathogens, and the evaluation of diagnostic techniques. Assessment of the phenotypic and genetic diversity in foodborne pathogens recovered in the frame of surveillance and monitoring programs Detection of factors associated with the dissemination of emergent strains/phenotypes/genotypes in the animal reservoir Reconstruction of the phylogeny of emergent strains/phenotypes/genotypes of Salmonella enterica, Campylobacter spp. y Escherichia coli

Evaluation of the performance of diagnostic tests for diseases of importance in animal health and assessment of their application in the frame of control and surveillance programs

Development and application of quantitative tools to the study the epidemiology of important diseases in public and animal health, including the evaluation of control and eradication programs of animal diseases, the spatial analysis, the molecular epidemiology and the identification of risk factors related to human and animal pathogens, and the evaluation of diagnostic techniques. Assessment of the sensitivity and specificity of diagnostic tests of routine use and new development using techniques not based on a gold reference (Bayesian latent class models) Design of protocols for the optimization of diagnostic strategies through the modification of cut-offs and other factors of the techniques Determination of the reliability of control, eradication and surveillance programs for animal pathogens (probability of detection at the group level, probability of disease freedom, estimating prevalence-adjusted estimates, etc.)

Antimicrobial resistances in pathogens, food-borne zoonotic agents and indicators

Annual surveillance programs in Antimicrobial resistance are established. Microorganisms are obtained in active sampling conducted in collaboration with the Central Administration, Regional Administration and pharmaceutical companies. Clinical isolates are performed with bacterial collections. Likewise, research in wild animals, urban effluents and clinical isolates was carried out. Evaluate the antimicrobial resistance profiles and their evolution in microorganisms isolated in food animals and food from animal origin and their relation with antimicrobial use. Identify the presence of antimicrobial resistant bacteria in wild animals and urban effluents. Studies of safety and efficacy of antimicrobials intended for use by animals. Study the temporal dynamics of resistant bacteria in laying hens.

Foodborne zoonoses in animals and meat, eggs and other products from animal origin intended for human consumption, mainly Salmonella spp. Campylobacter spp., Verotoxigenic Escherichia coli and Yersinia enterocolitica

Annual surveillance programs of zoonoses agents are established. Microorganisms are obtained in active sampling conducted in collaboration with different Administrations. Besides, research programs are performed to improve control programs of zoonotic agents. Additionally, the presence of these microorganisms is evaluated in wild animals or urban effluents samples. Analyze the prevalence of zoonotic agents in food animals and food from animal origin. Evaluate the presence of zoonotic agents in wild animals and environmental samples. Improve control programs of food-borne zoonoses. Evaluate different detection methods for the correct identification of foodborne pathogens in various matrices.

Genetic characterization of microorganisms populations

Different isolates are compared by genetic techniques (Pulsed Field Gel Electrophoresis, spa-typing, Multi Locus Sequence Typing,Next Generation Sequencing, etc.). Establish epidemiological links between microorganisms detected in animals, food and man, including samples of the environment and wildlife.

Genetic characterization of antimicrobial resistance

Isolates that show a resistance phenotype of interest to public health or animal health, are analyzed in depth using molecular techniques (molecular detection, Next Generation Sequencing, etc.). Linking antimicrobial resistance seen in animals, food and man, including samples of the environment and wildlife. Identify new mechanisms associated with resistance to major antibiotics used in human or veterinary medicine. Evaluate the use of new techniques for detection of antimicrobial resistance mechanisms in our isolates.

Molecular characterization of human reservoirs potentially pathogenic Escherichia coli, animal and environmental

Characterization of potentially pathogenic E. coli samples from different origins. Detection of virulence genes using quantitative real time PCR to distinguish the samples intended for isolation of a positive strain. Studies of antibiotic resistance in strains from healthy animals. Assess the genetic diversity of STEC isolates by pulsed field gel electrophoresis (PFGE) and Next Generation Sequencing (NGS). Detection serogroups / serotypes related-genes associated with hemolytic uremic syndrome (HUS). Shigatoxin producing E. coli prevalence studies in healthy animals.

Genetic characterization of antimicrobial resistance

Isolates that show a resistance phenotype of interest to public health or animal health, are analyzed in depth using molecular techniques (molecular detection, Next Generation Sequencing, etc.). Linking antimicrobial resistance seen in animals, food and man, including samples of the environment and wildlife. Identify new mechanisms associated with resistance to major antibiotics used in human or veterinary medicine. Evaluate the use of new techniques for detection of antimicrobial resistance mechanisms in our isolates.

Molecular characterization of human reservoirs potentially pathogenic Escherichia coli, animal and environmental

Characterization of potentially pathogenic E. coli samples from different origins. Detection of virulence genes using quantitative real time PCR to distinguish the samples intended for isolation of a positive strain. Studies of antibiotic resistance in strains from healthy animals. Assess the genetic diversity of STEC isolates by pulsed field gel electrophoresis (PFGE) and Next Generation Sequencing (NGS). Detection serogroups / serotypes related-genes associated with hemolytic uremic syndrome (HUS). Shigatoxin producing E. coli prevalence studies in healthy animals.

Genetic characterization of Campylobacter

Genetic characterization of Campylobacter jejuni and coli strains from different origins (animal, food and urban effluents). Genetic recombination between Campylobacter isolates: antibiotic resistance genes and virulence genes / colonization. Analysis of antimicrobial resistance in Campylobacter isolates from humans, animals and environment. Genetic diversity using whole-genome MLST in isolates from humans, animals and environment.

Modification of intestinal microbiota, Healthy Gut

An approach based on the use of natural compounds for the control of zoonotic or resistant bacteria. Study the intestinal microbiota present in food animals before and after the application of a polyphenols treatment. Nutrigenomics studies to identify changes in the expression of certain genes after treatment administration. Modify the intestinal microbiota, favoring the establishment and maintenance of a healthy and balanced intestinal bacterial population (lactic bacteria and bifidobacteria).after treatment administration. Control of enteric diseases and transmission of resistant bacteria in animal production (poultry, fish, ruminant and monogastric) after reduction of enterobacteria and resistant bacteria.

Marine animal health

The project Marine Animal Health has as main objectives the evaluation, monitoring and improving of health in the marine fauna of the Mediterranean Sea, contributing to conservation of the marine ecosystem. Assessment of health status in the marine fauna of the Mediterranean Sea. Surveillance of infectious diseases involved. Epidemiological studies on the definition of these diseases.

Preventive medicine

Development of different epidemiological models on the risks of entry different infectious diseases in our country, by identifying the critical points of each disease and modelling their potential spread. The diseases being studied are classified in the list of notifiable to the World Organization for Animal Health (OIE). To date we have developed a model for Bluetongue and others to foot and mouth disease, clasical swine fever and avian influenza. Risk analysis and identification of critical points at the entrance of infectious diseases in Spain. Development of epidemiological models that allow early detection of these diseases and the creation of proper contingency plans.

New diagnostic methods design

Development of new dignostic molecular methods based in the Methagenomics and PCR metodologies multiple and quantitative for several animal infectius deseases, in order to be able to carry out molecular epidemiology studies and develop techniques to differentiate vaccinated animals from infected. Incorporation of a new line of thermal imaging diagnosis to detect feverish conditions at an early stage and without the need for any manipulation of the animal. Evaluation of the system in different domestic and wild animals in both terrestrial and aquatic Studies of new animal viruses by metagenomics. Development of new diagnostic methods for viral diseases basads in the PCR. Epidemiological studies and development of design methods DIVA. Application of thermography in the study of infectious diseases.

Proliferative and non proliferative lesions of the gastrointestinal tract in small animals

Our participation in Gastrointestinal Course supervised by Dr. I. van der Gaag and Liver Pathology leaded by T. van den Ingh at the University of Utrecht (Netherlands), we started in this line of research. As a resulting investigations have been realized three PhD dissertations and numerous scientific articles in the JCR and communications at conferences national and international. Our research has focused on the differentiation of inflammatory disorders versus lymphosarcoma by means the use of immunohistochemical and molecular techniques. We have also made retrospective studies on epithelial lesions of the stomach and intestine, with special emphasis on cell cycle proteins which could be altered or participate in the transformation and expansion of benign or malignant neoplasms in dogs and cats. 1) Study of cell cycle oncoproteins involved in the development of neoplasms in the stomach and small intestine of pets. 2) Identification of early immunohistochemical biomarkers caracterizen passing a benign lesion to malignant. Prognosis. 3) Histological, immunohistochemical and molecular characterization of chronic enteritis linfoplasmocitaria digestive versus lymphoma.

Immunopathology of tuberculosis in domestic and wild animals

Our colaboration in European in the pathological aspects with Reference Laboratory on Bovine Tuberculosis began our journey in this research. Since then I have worked in the description of many pathological processes induced by different strains of Mycobacterium, both domestic and wild animals. This has led to PhD theses, dissertations, as well as many oral presentations and scientific articles on all related to the immune response to tissue level against different cytokines. 1) Study of the immune response in natural and experimental models in domestic animals with different strains of Mycobacterium. 2) Immune response in camels. 3) Tuberculosis in wild and zoo animals.

Fish pathology

In recent years, parallel to the development of aquaculture and intensive production has been a rise in infectious diseases, some of which have been major losses. Our work in this field has been to make a correct diagnostic pathology and microbiological isolation to take appropriate preventive measures. It is therefore applied research. Sometimes, these cases are the starting point for further investigations in the field of disease pathogenesis both conventional and specific techniques such as immunohistochemical, molecular and / or MALDI TOF methods. We also collaborate with other centers in carrying out histopathological studies on animal welfare and opportunistic pathogens in aquaculture. 1) Description of injuries caused by an infectious agent in an experimental studies on fish as well as its distribution in the body by means the use of monoclonal antibodies with immunohistochemical techniques. 2) Pathological characterization of infectious diseases in marine aquaculture.

Forensic Veterinary

As part of the Veterinary Pathology, increasingly there are many cases of animal abuse and natural disasters that require accurate and specific description of the process. For this reason, in recent years we have focused in particular on developing a specific methodology, which departs from the routine necropsy can be performed on a cadaver. Thus, it is necessary to conduct a detailed study of the external survey, sometimes for traumatic injuries, hangings, shootings, etc. 1) Development of methods for describing specific study of these cases, not only on the cause of death but also the elapsed playing time. 2) Establish a specific report and diagnostic protocol in Veterinary Forensic, which also includes macroscopic and histopathology descriptions, so as complementary toxicological and microbiological studies.

Malditof profiling and imaging

The first applications of mass spectrometry in clinical were as a method adapted to highly complex protein sample analysis in laboratories pathobiology, using high levels of sensitivity and specificity in the diagnosis of tumor processes, and other hereditary disorders markers of pathology. Today, one of the main applications of MALDI is bacterial identification in microbiology laboratories (MALDI Biotyoper), which has established itself as a fast and efficient analytical method has allowed the characterization of a large collection of microorganisms importance in the field of clinical microbiology (Clark et al., 2013). Moreover, the MALDI Imaging technology has become a powerful tool for molecular analysis of tissue sections (histological preparations), providing information on the spatial distribution and relative abundance of different components in these sections. This technology is very advantageous because it requires no specific target markers (such as immunohistochemical techniques), can detect in situ proteins, peptides, drugs and metabolites, has high sensitivity and high performance (Deutskens et al., 2011). Finally, in the MALDI Profiling identifying biomolecules (mostly peptide and protein nature) is carried out on biological fluids such as serum, urine and tissue extracts. 1) Application of MALDITOF techniques to the Veterinary Pathology. 2) Using the MALDITOF profiling in the identification of proteins as early biomarkers of joint disease in domestic animals. 3) Study of protein profile in canine digestive tumors by means MALDI Imaging technique, to establish premature protein changes as markers for malignant transformation.

Epidemiological surveillance of zoonotic tick-borne pathogens

Identification of ticks from domestic animals and wildlife and molecular detection of pathogens (genus Erlichia, Borrelia, Anaplasma, Francisella, Coxiella, Rickettsia y Bartonella) by PCR. Use of molecular techniques for identification of tick-borne pathogens at the species-level Identification of tick species recovered from different epidemiological contexts. Surveillance of tick-borne pathogens in ticks from wild and domestic animals. Implementation of molecular tools for identification of the bacterial species present in ticks.

New strategies for diagnosis and control of bovine, ovine and caprine brucellosis

Analysis of new approaches for the control of animal brucellosis using available vaccines, and evaluation of current strategies for disease control. Development of new tools for early detection of the pathogen using molecular techniques. Development of new routes of vaccination for control of small ruminants brucellosis Analysis of vaccination strategies for control of animal brucellosis Development of tools for detection and quantification of Brucella spp. in clinical samples

Diagnosis of swine brucellosis

Use of immunological diagnostic tests (detection of specific antibodies using rose bengale test and ELISA, experimental use of interferon-gamma detection assays) and bacteriology for the detection of infections caused by Brucella suis in swine and in wild boar. Etiological and epidemiological diagnosis of infections caused by Brucella suis in swine Optimization of techniques aiming at the detection of the cellular immune response for specific diagnosis of swine brucellosis Detection of Brucella suis in wild reservoir Implementation of molecular characterization techniques for the epidemiological analysis of outbreaks due to Brucella suis.

Diagnosis of Q fever in livestock and wildlife using direct and indirect tests

Analysis of clinical samples by means of direct (real-time PCR direct detection) and indirect (serological tests, IFN-gamma detection assay) diagnostic techniques to assess the distribution of Coxiella burnetii on the animal reservoir and to determine the immune response induced in infected animals. Assessment of the distribution of Coxiella burnetii in livestock and wildlife and the potential zoonotic risk as source of infection posed by the animal reservoir. Detection of risk factors associated with a higher infection risk in domestic ruminants. Development of indirect diagnostic tools for early diagnosis of C. burnetii infection.

Diagnosis of leishmaniasis in the animal reservoir

Research on the epidemiology of leishmaniasis in peri-urban settings. Identification of potential atypical reservoirs of Leishmania. Diagnosis of infection using indirect (indirect inmunofluorescence assay) and direct (PCR) techniques in biological samples from different hosts and vectors. Determination of the prevalence of leishmaniasis in animal reservoirs using serological and molecular techniques Analysis of potential risk/protective factors associated with the presence of the parasite Set up of new diagnostic tools Molecular characterization of L. infantum isolates

Integrated monitoring and control of Toxoplasma gondii and Hepatitis E virus in food supply chains

Research focused on characterizing the presence of emerging food-borne pathogens (in particular Toxoplasma gondii and hepatitis E virus) along the food chain, from the origin to the point of sale of the product. This researcher line will provide a new framework for the monitoring, tracking and modelling of health risks associated with these microorganisms, and will develop new food control procedures throughout the food chain. Harmonization of the methodology for the detection and characterization of Toxoplasma gondii and hepatitis E virus. Detection and characterization of Toxoplasma gondii and hepatitis E virus in wildlife. Evaluation of the risk of survival of Toxoplasma gondii and hepatitis E virus in different stages of the pig production chain: farm, slaughterhouse and point of sale of meat products available for consumption. Proposal for corrective measures and control methods for the management of risk associated with meat products.

Diagnosis of other mycobacteriosis

Diagnosis of other mycobacteria not included in the complexes M. tuberculosis and M. avium.

Molecular epidemiology of tubeculosis

Epidemiological studies based on molecular characterization data of M. tuberculosis complex members. - Molecular characterization of M. tuberculosis complex members. - Manteinance and update of the Spanish Database of Animal Mycobacteriosis mycoDB. - Clonal complexity studies.

Molecular epidemiology of tubeculosis

Epidemiological studies based on molecular characterization data of M. tuberculosis complex members. - Molecular characterization of M. tuberculosis complex members. - Manteinance and update of the Spanish Database of Animal Mycobacteriosis mycoDB. - Clonal complexity studies.

Tuberculosis diagnosis

Improvement and optimization of techniques for the diagnosis of tuberculosis in cattle and other animal species. Study of the immune response against tuberculosis. Sensitivity and specificity studies of the techniques used for the diagnosis of bovine tuberculosis. Studies of diagnosis interference in cellular and humoral immune response tests. Study of the immune response against tuberculosis. Development and set-up of new diagnostic techniques in cattle and other animal species. Cellular infection models to perform pathogenicity and virulence studies.

Tuberculosis control

Experimental and field studies using vaccination for the control of tuberculosis in domestic and wild animals. Vaccination studies as an alternative to the control of tuberculosis in wild animals (wild boar) and goats.

Animal Health

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