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Lorenzo Fraile

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DVM and PhD in Pharmacology at the Veterinary Faculty of Zaragoza, Spain. I worked as a swine and bovine practitioner for eight years.I worked as a senior researcher at CReSA (Center of Animal Health Research) working in the fields of epidemiology, pharmacology and immunology for seven years.From 2010 to now, I work as associate professor at the University of Lleida, Spain.During my career, I have published more than eighty-two papers in peer-review journals.

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Papers by Lorenzo Fraile

The Impact of Producing Type and Dietary Crude Protein on Animal Performances and Microbiota Together with Greenhouse Gases Emissions in Growing Pigs

Animals

In order to reduce dietary nitrogen and achieve an efficient protein deposition as well as decrea... more In order to reduce dietary nitrogen and achieve an efficient protein deposition as well as decrease N wastage, we challenged the nutrient utilization efficiency of two different producing types in front of a dietary crude protein (CP) restriction and studied the role of the microbiota in such an adaptation process. Therefore, 32 pure castrated male Duroc (DU) and 32 entire male hybrid (F2) piglets were raised in a three-phase feeding regime. At each phase, two iso caloric diets differing in CP content, also known as normal protein (NP) and low protein (LP), were fed to the animals. LP diets had a fixed restriction (2%) in CP content in regards to NP ones throughout the phases of the experiment. At the end of third phase, fecal samples were collected for microbiota analysis purposes and greenhouse gases emissions, together with ammonia, were tested. No changes were found in average daily feed intake (ADFI) of animals of two producing types (Duroc vs. F2) or those consumed different e...

Antimicrobial Resistance Genes in Porcine Pasteurella Multocida are not Associated with its Antimicrobial Susceptibility Pattern

Forty-eight Pasteurella multocida isolates were recovered from porcine pneumonic lungs collected ... more Forty-eight Pasteurella multocida isolates were recovered from porcine pneumonic lungs collected in Norwestern Spain (2017- 2019). These isolates were characterized for their minimal inhibition concentrations to twelve antimicrobial agents and for the appearance of eight resistance genes: tetA, tetB, blaROB1, blaTEM, ermA, ermC, mphE and msrE. Relevant resistance percentages were shown to teracyclines, sulphamethoxazole/trimethoprim and tiamulin, thus suggesting that P. multocida isolates were mostly susceptible to amoxicillin, ceftiofur, enrofloxacin, florfenicol, marbofloxacin and macrolides. 29.2% of isolates were resistant to more than two antimicrobials. The tetracycline resistance genes (tetA and tetB) were detected in 22.9% of the isolates, but none was positive to both simultaneously; blaROB1 and blaTEM genes were found in one third of isolates but both genes were detected simultaneously in only one isolate. ermC gene was observed in 41.7% of isolates, a percentage that decr...

Altered Nasal Microbiota Composition Associated with Development of Polyserositis by Mycoplasma hyorhinis

Pathogens

Fibrinous polyserositis in swine farming is a common pathological finding in nursery animals. The... more Fibrinous polyserositis in swine farming is a common pathological finding in nursery animals. The differential diagnosis of this finding should include Glaesserella parasuis (aetiological agent of Glässer’s disease) and Mycoplasma hyorhinis, among others. These microorganisms are early colonizers of the upper respiratory tract of piglets. The composition of the nasal microbiota at weaning was shown to constitute a predisposing factor for the development of Glässer’s disease. Here, we unravel the role of the nasal microbiota in the subsequent systemic infection by M. hyorhinis, and the similarities and differences with Glässer’s disease. Nasal samples from farms with recurrent problems with polyserositis associated with M. hyorhinis (MH) or Glässer’s disease (GD) were included in this study, together with healthy control farms (HC). Nasal swabs were taken from piglets in MH farms at weaning, before the onset of the clinical outbreaks, and were submitted to 16S rRNA gene amplicon sequ...

Modeling of Vaccination and Contact Tracing as Tools to Control the COVID-19 Outbreak in Spain

Vaccines

We developed an agent-based stochastic model, based on P Systems methodology, to decipher the eff... more We developed an agent-based stochastic model, based on P Systems methodology, to decipher the effects of vaccination and contact tracing on the control of COVID-19 outbreak at population level under different control measures (social distancing, mask wearing and hand hygiene) and epidemiological scenarios. Our findings suggest that without the application of protection social measures, 56.1% of the Spanish population would contract the disease with a mortality of 0.4%. Assuming that 20% of the population was protected by vaccination by the end of the summer of 2021, it would be expected that 45% of the population would contract the disease and 0.3% of the population would die. However, both of these percentages are significantly lower when social measures were adopted, being the best results when social measures are in place and 40% of contacts traced. Our model shows that if 40% of the population can be vaccinated, even without social control measures, the percentage of people who ...

The Specific Immune Response after Vaccination against Neonatal Calf Diarrhoea Differs between Apparent Similar Vaccines in a Case Study

Animals

Neonatal calf diarrhoea (NCD) is a major health challenge with a negative impact on farm profitab... more Neonatal calf diarrhoea (NCD) is a major health challenge with a negative impact on farm profitability, calf welfare and antimicrobial use. Neonatal calves are particularly sensitive to enteric infections. Thus, a key point for prevention is minimising infectious pressure and maximising specific immune responses. An amount of 120 dams not previously vaccinated against NCD were randomly allocated to one of three study groups: negative control versus two vaccinated groups (A and B). In the control group, the average level of antibodies was significantly low for both BoCV and ETEC (15.6 and 13.9% in the colostrum samples, respectively), demonstrating the importance of dam vaccination. Indeed, the level of specific immunity was significantly increased for BoCV and ETEC with dam vaccination using both one-shot vaccines versus the control group. Moreover, the statistical analysis revealed a significantly higher level of antibodies for BoCV and ETEC in colostrum samples in vaccine A versus...

Vaccination Is a Suitable Tool in the Control of Aujeszky’s Disease Outbreaks in Pigs Using a Population Dynamics P Systems Model

Animals

Aujeszky’s disease is one of the main pig viral diseases and results in considerable economic los... more Aujeszky’s disease is one of the main pig viral diseases and results in considerable economic losses in the pork production industry. The disease can be controlled using preventive measures such as improved stock management and vaccination throughout the pig-rearing period. We developed a stochastic model based on Population Dynamics P systems (PDP) models for a standard pig production system to differentiate between the effects of pig farm management regimes and vaccination strategies on the control of Aujeszky’s disease under several different epidemiological scenarios. Our results suggest that after confirming the diagnosis, early vaccination of most of the population (>75%) is critical to decrease the spread of the virus and minimize its impact on pig productivity. The direct economic cost of an outbreak of Aujeszky’s disease can be extremely high on a previously uninfected farm (from 352–792 Euros/sow/year) and highlights the positive benefits of investing in vaccination mea...

Resilience Effects of SGK1 and TAP1 DNA Markers during PRRSV Outbreaks in Reproductive Sows

Animals

The porcine reproductive and respiratory syndrome virus (PRRSV) is a major infectious stressor th... more The porcine reproductive and respiratory syndrome virus (PRRSV) is a major infectious stressor that causes serious health problems and productivity drops. Based on previous genome-wide analyses, we selected SGK1 and TAP1 as candidate genes for resilience, and genotyped three mutations, including a 3′UTR variant SGK1_rs338508371 and two synonymous variants TAP1_rs1109026889 and TAP1_rs80928141 in 305 Landrace × Large White sows. All polymorphisms affected the reproductive performance in the outbreak, but not during the endemic phase, thereby indicating a potential use of these markers for resilience. Moreover, some genotypes were associated with a stable performance across PRRSV phases. Thus, in the outbreak, the SGK1_rs338508371 AA sows had less piglets born alive (p < 0.0001) and more stillborns (p < 0.05) while other sows were able to keep their productivity. During the outbreak, TAP1_rs80928141 GG sows had less piglets born alive (p < 0.05) and both TAP1 polymorphisms in...

Identification of resilient sows in Porcine Reproductive and Respiratory Syndrome virus infected farms

Journal of Animal Science

The identification of resilient sows can improve reproductive performance in farms exposed to mul... more The identification of resilient sows can improve reproductive performance in farms exposed to multiple challenges. A common challenge is the porcine reproductive and respiratory syndrome virus (PRRSV). A key issue to deal with disease resilience is to set up a feasible phenotyping strategy. Our aim was to develop a phenotyping criterion to discriminate susceptible from resilient sows in PRRSV-infected farms. A total of 517 Landrace x Large White gilts were classified as resilient (R) or susceptible (S) to PRRSV virus, following vaccination with MLV-PRRSV at 6 to 7 wk of age, in a PRRSV negative multiplication farm. Female piglets were phenotyped as R if their serum was negative to PRRSV at 7 and 21 d postvaccination (DPV) or as S if their serum was positive at 7 and/or 21 DPV. Amongst them, 382 gilts were transferred to a PRRSV-positive production farm, where the number of piglets born alive (NBA), stillborn (NSB), mummified (NMU), lost (NLP = NSB + NMU), and total born (NTB = NBA +...

Cellular Innate Immunity against PRRSV and Swine Influenza Viruses

Veterinary Sciences

Porcine respiratory disease complex (PRDC) is a polymicrobial syndrome that results from a combin... more Porcine respiratory disease complex (PRDC) is a polymicrobial syndrome that results from a combination of infectious agents, such as environmental stressors, population size, management strategies, age, and genetics. PRDC results in reduced performance as well as increased mortality rates and production costs in the pig industry worldwide. This review focuses on the interactions of two enveloped RNA viruses—porcine reproductive and respiratory syndrome virus (PRRSV) and swine influenza virus (SwIV)—as major etiological agents that contribute to PRDC within the porcine cellular innate immunity during infection. The innate immune system of the porcine lung includes alveolar and parenchymal/interstitial macrophages, neutrophils (PMN), conventional dendritic cells (DC) and plasmacytoid DC, natural killer cells, and γδ T cells, thus the in vitro and in vivo interactions between those cells and PRRSV and SwIV are reviewed. Likewise, the few studies regarding PRRSV-SwIV co-infection are il...

Key Gaps in the Knowledge of the Porcine Respiratory Reproductive Syndrome Virus (PRRSV)

Frontiers in Veterinary Science

$Research paper thumbnail of Targeted-pig trial on safety and immunogenicity of serum-derived extracellular vesicles enriched fractions obtained from Porcine Respiratory and Reproductive virus infections$

Targeted-pig trial on safety and immunogenicity of serum-derived extracellular vesicles enriched fractions obtained from Porcine Respiratory and Reproductive virus infections

Scientific Reports

The Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is the etiological agent of one o... more The Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is the etiological agent of one of the most important swine diseases with a significant economic burden worldwide. Unfortunately, available vaccines are partially effective highlighting the need of novel approaches. Previously, antigenic viral proteins were described in serum-derived extracellular vesicles (EVs) from pigs previously infected with PRRSV. Here, a targeted-pig trial was designed to determine the safety and immunogenicity of such extracellular vesicles enriched fractions. Our results showed that immunizations with EV-enriched fractions from convalescence animals in combination with montanide is safe and free of virus as immunizations with up-to two milligrams of EV-enriched fractions did not induce clinical symptoms, adverse effects and detectable viral replication. In addition, this vaccine formulation was able to elicit specific humoral IgG immune response in vaccinated animals, albeit variably. Noticeably, sera from vaccinated animals was diagnosed negative when tested for PRRSV using a commercial ELISA test; thus, indicating that this new approach differentiates vaccinated from infected animals. Lastly, after priming animals with EV-enriched fractions from sera of convalescence animals and boosting them with synthetic viral peptides identified by mass spectrometry, a distinctive high and specific IFN-γ response was elicited. Altogether, our data strongly suggest the use of serum EV-enriched fractions as a novel vaccine strategy against PRRSV. Recent estimates calculate that the world human population will reach near 9,6 billion people by 2050 and the United Nations Food and Agriculture Organization (FAO) estimates that in order to feed this population the overall food production will need to increase by 70% 1. Undoubtedly, veterinary vaccines that preserve animal health and improve production will play an essential role in reaching this goal. Moreover, they will reduce the use of antibiotics as they are escalating into a global health crisis. To reach this goal, novel vaccination approaches are desperately required as classical and live-attenuated vaccines are far from being totally safe and efficacious in animal diseases 2. The Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) causes one of the most important diseases of veterinary medicine 3. Recent economical estimates in the United States, where the disease is highly prevalent, indicate more than 600 million $ loses each year 4. Current vaccines against PRRSV use modified live or

Multidrug resistant Salmonella enterica isolated from conventional pig farms using antimicrobial agents in preventative medicine programmes

Veterinary journal (London, England : 1997), Apr 1, 2018

A longitudinal study was conducted to investigate the presence of multidrug antimicrobial resista... more A longitudinal study was conducted to investigate the presence of multidrug antimicrobial resistance (multi-AR) in Salmonella enterica in pigs reared under conventional preventative medicine programmes in Spain and the possible association of multi-AR with ceftiofur or tulathromycin treatment during the pre-weaning period. Groups of 7-day-old piglets were treated by intramuscular injection with ceftiofur on four farms (n=40 piglets per farm) and with tulathromycin on another four farms (n=40 piglets per farm). A control group of untreated piglets (n=30 per farm) was present on each farm. Faecal swabs were collected for S. enterica culture prior to treatment, at 2, 7 and 180days post-treatment, and at slaughter. Minimal inhibitory concentrations of 14 antimicrobial agents, pulsed-field gel electrophoresis and detection of resistance genes representing five families of antimicrobial agents were performed. Plasmids carrying cephalosporin resistant (CR) genes were characterised. Sixty-s...

A bivalent dendrimeric peptide bearing a T-cell epitope from foot-and-mouth disease virus protein 3A improves humoral response against classical swine fever virus

Virus Research

Està subjecte a una llicència de Reconeixement-NoComercial-SenseObraDerivada 4.

African swine fever virus infection in Classical swine fever subclinically infected wild boars

BMC veterinary research, 2017

Recently moderate-virulence classical swine fever virus (CSFV) strains have been proven capable o... more Recently moderate-virulence classical swine fever virus (CSFV) strains have been proven capable of generating postnatal persistent infection (PI), defined by the maintenance of viremia and the inability to generate CSFV-specific immune responses in animals. These animals also showed a type I interferon blockade in the absence of clinical signs. In this study, we assessed the infection generated in 7-week-old CSFV PI wild boars after infection with the African swine fever virus (ASFV). The wild boars were divided in two groups and were infected with ASFV. Group A comprised boars who were CSFV PI in a subclinical form and Group B comprised pestivirus-free wild boars. Some relevant parameters related to CSFV replication and the immune response of CSFV PI animals were studied. Additionally, serum soluble factors such as IFN-α, TNF-α, IL-6, IL-10, IFN-γ and sCD163 were analysed before and after ASFV infection to assess their role in disease progression. After ASFV infection, only the CSF...

Potential use of local and systemic humoral immune response parameters to forecast Mycoplasma hyopneumoniae associated lung lesions

PLOS ONE, 2017

Immunopathological events are key for the development of enzootic pneumonia (EP), which is macros... more Immunopathological events are key for the development of enzootic pneumonia (EP), which is macroscopically observed as cranioventral pulmonary consolidation (CVPC). This study aimed to investigate the putative association between the humoral immune response against Mycoplasma hyopneumoniae (M. hyopneumoniae) and prevalence and extension of CVPC in 1) experimentally infected pigs, 2) slaughtered pigs and 3) sequentially necropsied pigs in a longitudinal study. CVPC was scored by means of the European Pharmacopoeia recommended methodology. Specific IgG, IgG1 and IgG2 antibodies were assessed in serum. In addition, mucosal IgG and IgA antibodies were analyzed in broncho-alveolar lavage fluid (BALF) from experimentally challenged pigs. The systemic humoral immune response in experimentally infected pigs was delayed in onset whereas humoral respiratory mucosal immune response appeared more rapidly but declined earlier. Although low, BALF IgG antibodies showed the highest correlation with CVPC scores (r = 0.49, p<0.05). In slaughter-aged pigs, both percentage of lungs with CVPC and mean lung lesion score were significantly higher in M. hyopneumoniae seropositive farms compared to the seronegative ones (p<0.001). Similarly, seropositive sequentially necropsied pigs showed more severe CVPC than seronegative ones. Overall, mean serological values might help to forecast prevalence and severity of EP-like lung lesions using a population based approach. Remarkably, the specific systemic humoral immune response was found to be predominated by the IgG2 subclass, suggesting a dominant Th1mediated immune response to M. hyopneumoniae.

Biochemical and proteomic analyses of the physiological response induced by individual housing in gilts provide new potential stress markers

BMC Veterinary Research, 2016

Background: The objective assessment of animal stress and welfare requires proper laboratory biom... more Background: The objective assessment of animal stress and welfare requires proper laboratory biomarkers. In this work, we have analyzed the changes in serum composition in gilts after switching their housing, from pen to individual stalls, which is generally accepted to cause animal discomfort. Results: Blood and saliva samples were collected a day before and up to four days after changing the housing system. Biochemical analyses showed adaptive changes in lipid and protein metabolism after the housing switch, whereas cortisol and muscular markers showed a large variability between animals. 2D-DIGE and iTRAQ proteomic approaches revealed variations in serum protein composition after changing housing and diet of gilts. Both techniques showed alterations in two main homeostatic mechanisms: the innate immune and redox systems. The acute phase proteins haptoglobin, apolipoprotein A-I and α1-antichymotrypsin 3, and the antioxidant enzyme peroxiredoxin 2 were found differentially expressed by 2D-DIGE. Other proteins related to the innate immune system, including lactotransferrin, protegrin 3 and galectin 1 were also identified by iTRAQ, as well as oxidative stress enzymes such as peroxiredoxin 2 and glutathione peroxidase 3. Proteomics also revealed the decrease of apolipoproteins, and the presence of intracellular proteins in serum, which may indicate physical injury to tissues. Conclusions: Housing of gilts in individual stalls and diet change increase lipid and protein catabolism, oxidative stress, activate the innate immune system and cause a certain degree of tissue damage. We propose that valuable assays for stress assessment in gilts may be based on a score composed by a combination of salivary cortisol, lipid metabolites, innate immunity and oxidative stress markers and intracellular proteins.

Infection of dendritic cells with the American, but not the European, PRRSv genotype induces Treg (Foxp3+CD25+) cells

The Faseb Journal, Mar 1, 2008

Piglet nasal microbiota at weaning may influence the development of Glässer’s disease during the rearing period

BMC Genomics, 2016

Background: The microbiota, the ensemble of microorganisms on a particular body site, has been ex... more Background: The microbiota, the ensemble of microorganisms on a particular body site, has been extensively studied during the last few years, and demonstrated to influence the development of many diseases. However, these studies focused mainly on the human digestive system, while the populations in the respiratory tract have been poorly assessed, especially in pigs. The nasal mucosa of piglets is colonized by an array of bacteria, many of which are unknown. Among the early colonizers, Haemophilus parasuis also has clinical importance, since it is also the etiological agent of Glässer's disease. This disease produces economical losses in all the countries with pig production, and the factors influencing its development are not totally understood. Hence, the purpose of this work was to characterize the nasal microbiota composition of piglets, and its possible role in Glässer's disease development. Results: Seven farms from Spain (4 with Glässer's disease and 3 control farms without any respiratory disease) and three farms from UK (all control farms) were studied. Ten piglets from each farm were sampled at 3-4 weeks of age before weaning. The total DNA extracted from nasal swabs was used to amplify the 16S RNA gene for sequencing in Illumina MiSeq. Sequencing data was quality filtered and analyzed using QIIME software. The diversity of the nasal microbiota was low in comparison with other body sites, showing a maximum number of operational taxonomic units (OTUs) per pig of 1,603, clustered in five phyla. Significant differences were found at various taxonomical levels, when the microbiota was compared regarding the farm health status. Healthy status was associated to higher species richness and diversity, and UK farms demonstrated the highest diversity. Conclusions: The composition of the nasal microbiota of healthy piglets was uncovered and different phylotypes were shown to be significantly altered in animals depending on the clinical status of the farm of origin. Several OTUs at genus level were identified over-represented in piglets from control farms, indicating their potential as probiotics. Although we provide relevant data, fully metagenomic approaches could give light on the genes and metabolic pathways involved in the roles of the nasal microbiota to prevent respiratory diseases.

Marbofloxacin reaches high concentration in pig tonsils in a dose-dependent fashion: Marbofloxacin reaches high concentration in pig tonsils

J Vet Pharmacol Ther, 2010

Shedding of cephalosporin resistant Escherichia coli in pigs from conventional farms after early treatment with antimicrobials

Veterinary journal (London, England : 1997), Jan 4, 2016

This study assessed the dynamics of cephalosporin resistant (CR) E. coli populations during the l... more This study assessed the dynamics of cephalosporin resistant (CR) E. coli populations during the life cycle of pigs treated early in life with ceftiofur or tulathromycin. The study was conducted at eight conventional pig farms; four for each treatment with ceftiofur or tulathromycin. At each farm, 70 7-day-old piglets were divided into two groups: a control group (n = 30) and a treatment group (n = 40). Faecal samples were collected on day 0 and on days 2, 7 and 180 post-treatment. Sows were also sampled on day 0. CR E. coli were selected on MacConkey agar with ceftriaxone. On five farms, 7-day-old piglets excreted CR E. coli before treatment associated with the presence of CR E. coli in sows. The occurrence of CR E. coli positive animals decreased with increasing piglet age. The remaining three farms tested negative for CR E. coli during the study period. Results demonstrated great variability in the frequency of CR E. coli positive animals between farms, independent of treatment. T...

The Impact of Producing Type and Dietary Crude Protein on Animal Performances and Microbiota Together with Greenhouse Gases Emissions in Growing Pigs

Animals

In order to reduce dietary nitrogen and achieve an efficient protein deposition as well as decrea... more In order to reduce dietary nitrogen and achieve an efficient protein deposition as well as decrease N wastage, we challenged the nutrient utilization efficiency of two different producing types in front of a dietary crude protein (CP) restriction and studied the role of the microbiota in such an adaptation process. Therefore, 32 pure castrated male Duroc (DU) and 32 entire male hybrid (F2) piglets were raised in a three-phase feeding regime. At each phase, two iso caloric diets differing in CP content, also known as normal protein (NP) and low protein (LP), were fed to the animals. LP diets had a fixed restriction (2%) in CP content in regards to NP ones throughout the phases of the experiment. At the end of third phase, fecal samples were collected for microbiota analysis purposes and greenhouse gases emissions, together with ammonia, were tested. No changes were found in average daily feed intake (ADFI) of animals of two producing types (Duroc vs. F2) or those consumed different e...

Antimicrobial Resistance Genes in Porcine Pasteurella Multocida are not Associated with its Antimicrobial Susceptibility Pattern

Forty-eight Pasteurella multocida isolates were recovered from porcine pneumonic lungs collected ... more Forty-eight Pasteurella multocida isolates were recovered from porcine pneumonic lungs collected in Norwestern Spain (2017- 2019). These isolates were characterized for their minimal inhibition concentrations to twelve antimicrobial agents and for the appearance of eight resistance genes: tetA, tetB, blaROB1, blaTEM, ermA, ermC, mphE and msrE. Relevant resistance percentages were shown to teracyclines, sulphamethoxazole/trimethoprim and tiamulin, thus suggesting that P. multocida isolates were mostly susceptible to amoxicillin, ceftiofur, enrofloxacin, florfenicol, marbofloxacin and macrolides. 29.2% of isolates were resistant to more than two antimicrobials. The tetracycline resistance genes (tetA and tetB) were detected in 22.9% of the isolates, but none was positive to both simultaneously; blaROB1 and blaTEM genes were found in one third of isolates but both genes were detected simultaneously in only one isolate. ermC gene was observed in 41.7% of isolates, a percentage that decr...

Altered Nasal Microbiota Composition Associated with Development of Polyserositis by Mycoplasma hyorhinis

Pathogens

Fibrinous polyserositis in swine farming is a common pathological finding in nursery animals. The... more Fibrinous polyserositis in swine farming is a common pathological finding in nursery animals. The differential diagnosis of this finding should include Glaesserella parasuis (aetiological agent of Glässer’s disease) and Mycoplasma hyorhinis, among others. These microorganisms are early colonizers of the upper respiratory tract of piglets. The composition of the nasal microbiota at weaning was shown to constitute a predisposing factor for the development of Glässer’s disease. Here, we unravel the role of the nasal microbiota in the subsequent systemic infection by M. hyorhinis, and the similarities and differences with Glässer’s disease. Nasal samples from farms with recurrent problems with polyserositis associated with M. hyorhinis (MH) or Glässer’s disease (GD) were included in this study, together with healthy control farms (HC). Nasal swabs were taken from piglets in MH farms at weaning, before the onset of the clinical outbreaks, and were submitted to 16S rRNA gene amplicon sequ...

Modeling of Vaccination and Contact Tracing as Tools to Control the COVID-19 Outbreak in Spain

Vaccines

We developed an agent-based stochastic model, based on P Systems methodology, to decipher the eff... more We developed an agent-based stochastic model, based on P Systems methodology, to decipher the effects of vaccination and contact tracing on the control of COVID-19 outbreak at population level under different control measures (social distancing, mask wearing and hand hygiene) and epidemiological scenarios. Our findings suggest that without the application of protection social measures, 56.1% of the Spanish population would contract the disease with a mortality of 0.4%. Assuming that 20% of the population was protected by vaccination by the end of the summer of 2021, it would be expected that 45% of the population would contract the disease and 0.3% of the population would die. However, both of these percentages are significantly lower when social measures were adopted, being the best results when social measures are in place and 40% of contacts traced. Our model shows that if 40% of the population can be vaccinated, even without social control measures, the percentage of people who ...

The Specific Immune Response after Vaccination against Neonatal Calf Diarrhoea Differs between Apparent Similar Vaccines in a Case Study

Animals

Neonatal calf diarrhoea (NCD) is a major health challenge with a negative impact on farm profitab... more Neonatal calf diarrhoea (NCD) is a major health challenge with a negative impact on farm profitability, calf welfare and antimicrobial use. Neonatal calves are particularly sensitive to enteric infections. Thus, a key point for prevention is minimising infectious pressure and maximising specific immune responses. An amount of 120 dams not previously vaccinated against NCD were randomly allocated to one of three study groups: negative control versus two vaccinated groups (A and B). In the control group, the average level of antibodies was significantly low for both BoCV and ETEC (15.6 and 13.9% in the colostrum samples, respectively), demonstrating the importance of dam vaccination. Indeed, the level of specific immunity was significantly increased for BoCV and ETEC with dam vaccination using both one-shot vaccines versus the control group. Moreover, the statistical analysis revealed a significantly higher level of antibodies for BoCV and ETEC in colostrum samples in vaccine A versus...

Vaccination Is a Suitable Tool in the Control of Aujeszky’s Disease Outbreaks in Pigs Using a Population Dynamics P Systems Model

Animals

Aujeszky’s disease is one of the main pig viral diseases and results in considerable economic los... more Aujeszky’s disease is one of the main pig viral diseases and results in considerable economic losses in the pork production industry. The disease can be controlled using preventive measures such as improved stock management and vaccination throughout the pig-rearing period. We developed a stochastic model based on Population Dynamics P systems (PDP) models for a standard pig production system to differentiate between the effects of pig farm management regimes and vaccination strategies on the control of Aujeszky’s disease under several different epidemiological scenarios. Our results suggest that after confirming the diagnosis, early vaccination of most of the population (>75%) is critical to decrease the spread of the virus and minimize its impact on pig productivity. The direct economic cost of an outbreak of Aujeszky’s disease can be extremely high on a previously uninfected farm (from 352–792 Euros/sow/year) and highlights the positive benefits of investing in vaccination mea...

Resilience Effects of SGK1 and TAP1 DNA Markers during PRRSV Outbreaks in Reproductive Sows

Animals

The porcine reproductive and respiratory syndrome virus (PRRSV) is a major infectious stressor th... more The porcine reproductive and respiratory syndrome virus (PRRSV) is a major infectious stressor that causes serious health problems and productivity drops. Based on previous genome-wide analyses, we selected SGK1 and TAP1 as candidate genes for resilience, and genotyped three mutations, including a 3′UTR variant SGK1_rs338508371 and two synonymous variants TAP1_rs1109026889 and TAP1_rs80928141 in 305 Landrace × Large White sows. All polymorphisms affected the reproductive performance in the outbreak, but not during the endemic phase, thereby indicating a potential use of these markers for resilience. Moreover, some genotypes were associated with a stable performance across PRRSV phases. Thus, in the outbreak, the SGK1_rs338508371 AA sows had less piglets born alive (p < 0.0001) and more stillborns (p < 0.05) while other sows were able to keep their productivity. During the outbreak, TAP1_rs80928141 GG sows had less piglets born alive (p < 0.05) and both TAP1 polymorphisms in...

Identification of resilient sows in Porcine Reproductive and Respiratory Syndrome virus infected farms

Journal of Animal Science

The identification of resilient sows can improve reproductive performance in farms exposed to mul... more The identification of resilient sows can improve reproductive performance in farms exposed to multiple challenges. A common challenge is the porcine reproductive and respiratory syndrome virus (PRRSV). A key issue to deal with disease resilience is to set up a feasible phenotyping strategy. Our aim was to develop a phenotyping criterion to discriminate susceptible from resilient sows in PRRSV-infected farms. A total of 517 Landrace x Large White gilts were classified as resilient (R) or susceptible (S) to PRRSV virus, following vaccination with MLV-PRRSV at 6 to 7 wk of age, in a PRRSV negative multiplication farm. Female piglets were phenotyped as R if their serum was negative to PRRSV at 7 and 21 d postvaccination (DPV) or as S if their serum was positive at 7 and/or 21 DPV. Amongst them, 382 gilts were transferred to a PRRSV-positive production farm, where the number of piglets born alive (NBA), stillborn (NSB), mummified (NMU), lost (NLP = NSB + NMU), and total born (NTB = NBA +...

Cellular Innate Immunity against PRRSV and Swine Influenza Viruses

Veterinary Sciences

Porcine respiratory disease complex (PRDC) is a polymicrobial syndrome that results from a combin... more Porcine respiratory disease complex (PRDC) is a polymicrobial syndrome that results from a combination of infectious agents, such as environmental stressors, population size, management strategies, age, and genetics. PRDC results in reduced performance as well as increased mortality rates and production costs in the pig industry worldwide. This review focuses on the interactions of two enveloped RNA viruses—porcine reproductive and respiratory syndrome virus (PRRSV) and swine influenza virus (SwIV)—as major etiological agents that contribute to PRDC within the porcine cellular innate immunity during infection. The innate immune system of the porcine lung includes alveolar and parenchymal/interstitial macrophages, neutrophils (PMN), conventional dendritic cells (DC) and plasmacytoid DC, natural killer cells, and γδ T cells, thus the in vitro and in vivo interactions between those cells and PRRSV and SwIV are reviewed. Likewise, the few studies regarding PRRSV-SwIV co-infection are il...

Key Gaps in the Knowledge of the Porcine Respiratory Reproductive Syndrome Virus (PRRSV)

Frontiers in Veterinary Science

$Research paper thumbnail of Targeted-pig trial on safety and immunogenicity of serum-derived extracellular vesicles enriched fractions obtained from Porcine Respiratory and Reproductive virus infections$

Targeted-pig trial on safety and immunogenicity of serum-derived extracellular vesicles enriched fractions obtained from Porcine Respiratory and Reproductive virus infections

Scientific Reports

The Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is the etiological agent of one o... more The Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is the etiological agent of one of the most important swine diseases with a significant economic burden worldwide. Unfortunately, available vaccines are partially effective highlighting the need of novel approaches. Previously, antigenic viral proteins were described in serum-derived extracellular vesicles (EVs) from pigs previously infected with PRRSV. Here, a targeted-pig trial was designed to determine the safety and immunogenicity of such extracellular vesicles enriched fractions. Our results showed that immunizations with EV-enriched fractions from convalescence animals in combination with montanide is safe and free of virus as immunizations with up-to two milligrams of EV-enriched fractions did not induce clinical symptoms, adverse effects and detectable viral replication. In addition, this vaccine formulation was able to elicit specific humoral IgG immune response in vaccinated animals, albeit variably. Noticeably, sera from vaccinated animals was diagnosed negative when tested for PRRSV using a commercial ELISA test; thus, indicating that this new approach differentiates vaccinated from infected animals. Lastly, after priming animals with EV-enriched fractions from sera of convalescence animals and boosting them with synthetic viral peptides identified by mass spectrometry, a distinctive high and specific IFN-γ response was elicited. Altogether, our data strongly suggest the use of serum EV-enriched fractions as a novel vaccine strategy against PRRSV. Recent estimates calculate that the world human population will reach near 9,6 billion people by 2050 and the United Nations Food and Agriculture Organization (FAO) estimates that in order to feed this population the overall food production will need to increase by 70% 1. Undoubtedly, veterinary vaccines that preserve animal health and improve production will play an essential role in reaching this goal. Moreover, they will reduce the use of antibiotics as they are escalating into a global health crisis. To reach this goal, novel vaccination approaches are desperately required as classical and live-attenuated vaccines are far from being totally safe and efficacious in animal diseases 2. The Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) causes one of the most important diseases of veterinary medicine 3. Recent economical estimates in the United States, where the disease is highly prevalent, indicate more than 600 million $ loses each year 4. Current vaccines against PRRSV use modified live or

Multidrug resistant Salmonella enterica isolated from conventional pig farms using antimicrobial agents in preventative medicine programmes

Veterinary journal (London, England : 1997), Apr 1, 2018

A longitudinal study was conducted to investigate the presence of multidrug antimicrobial resista... more A longitudinal study was conducted to investigate the presence of multidrug antimicrobial resistance (multi-AR) in Salmonella enterica in pigs reared under conventional preventative medicine programmes in Spain and the possible association of multi-AR with ceftiofur or tulathromycin treatment during the pre-weaning period. Groups of 7-day-old piglets were treated by intramuscular injection with ceftiofur on four farms (n=40 piglets per farm) and with tulathromycin on another four farms (n=40 piglets per farm). A control group of untreated piglets (n=30 per farm) was present on each farm. Faecal swabs were collected for S. enterica culture prior to treatment, at 2, 7 and 180days post-treatment, and at slaughter. Minimal inhibitory concentrations of 14 antimicrobial agents, pulsed-field gel electrophoresis and detection of resistance genes representing five families of antimicrobial agents were performed. Plasmids carrying cephalosporin resistant (CR) genes were characterised. Sixty-s...

A bivalent dendrimeric peptide bearing a T-cell epitope from foot-and-mouth disease virus protein 3A improves humoral response against classical swine fever virus

Virus Research

Està subjecte a una llicència de Reconeixement-NoComercial-SenseObraDerivada 4.

African swine fever virus infection in Classical swine fever subclinically infected wild boars

BMC veterinary research, 2017

Recently moderate-virulence classical swine fever virus (CSFV) strains have been proven capable o... more Recently moderate-virulence classical swine fever virus (CSFV) strains have been proven capable of generating postnatal persistent infection (PI), defined by the maintenance of viremia and the inability to generate CSFV-specific immune responses in animals. These animals also showed a type I interferon blockade in the absence of clinical signs. In this study, we assessed the infection generated in 7-week-old CSFV PI wild boars after infection with the African swine fever virus (ASFV). The wild boars were divided in two groups and were infected with ASFV. Group A comprised boars who were CSFV PI in a subclinical form and Group B comprised pestivirus-free wild boars. Some relevant parameters related to CSFV replication and the immune response of CSFV PI animals were studied. Additionally, serum soluble factors such as IFN-α, TNF-α, IL-6, IL-10, IFN-γ and sCD163 were analysed before and after ASFV infection to assess their role in disease progression. After ASFV infection, only the CSF...

Potential use of local and systemic humoral immune response parameters to forecast Mycoplasma hyopneumoniae associated lung lesions

PLOS ONE, 2017

Immunopathological events are key for the development of enzootic pneumonia (EP), which is macros... more Immunopathological events are key for the development of enzootic pneumonia (EP), which is macroscopically observed as cranioventral pulmonary consolidation (CVPC). This study aimed to investigate the putative association between the humoral immune response against Mycoplasma hyopneumoniae (M. hyopneumoniae) and prevalence and extension of CVPC in 1) experimentally infected pigs, 2) slaughtered pigs and 3) sequentially necropsied pigs in a longitudinal study. CVPC was scored by means of the European Pharmacopoeia recommended methodology. Specific IgG, IgG1 and IgG2 antibodies were assessed in serum. In addition, mucosal IgG and IgA antibodies were analyzed in broncho-alveolar lavage fluid (BALF) from experimentally challenged pigs. The systemic humoral immune response in experimentally infected pigs was delayed in onset whereas humoral respiratory mucosal immune response appeared more rapidly but declined earlier. Although low, BALF IgG antibodies showed the highest correlation with CVPC scores (r = 0.49, p<0.05). In slaughter-aged pigs, both percentage of lungs with CVPC and mean lung lesion score were significantly higher in M. hyopneumoniae seropositive farms compared to the seronegative ones (p<0.001). Similarly, seropositive sequentially necropsied pigs showed more severe CVPC than seronegative ones. Overall, mean serological values might help to forecast prevalence and severity of EP-like lung lesions using a population based approach. Remarkably, the specific systemic humoral immune response was found to be predominated by the IgG2 subclass, suggesting a dominant Th1mediated immune response to M. hyopneumoniae.

Biochemical and proteomic analyses of the physiological response induced by individual housing in gilts provide new potential stress markers

BMC Veterinary Research, 2016

Background: The objective assessment of animal stress and welfare requires proper laboratory biom... more Background: The objective assessment of animal stress and welfare requires proper laboratory biomarkers. In this work, we have analyzed the changes in serum composition in gilts after switching their housing, from pen to individual stalls, which is generally accepted to cause animal discomfort. Results: Blood and saliva samples were collected a day before and up to four days after changing the housing system. Biochemical analyses showed adaptive changes in lipid and protein metabolism after the housing switch, whereas cortisol and muscular markers showed a large variability between animals. 2D-DIGE and iTRAQ proteomic approaches revealed variations in serum protein composition after changing housing and diet of gilts. Both techniques showed alterations in two main homeostatic mechanisms: the innate immune and redox systems. The acute phase proteins haptoglobin, apolipoprotein A-I and α1-antichymotrypsin 3, and the antioxidant enzyme peroxiredoxin 2 were found differentially expressed by 2D-DIGE. Other proteins related to the innate immune system, including lactotransferrin, protegrin 3 and galectin 1 were also identified by iTRAQ, as well as oxidative stress enzymes such as peroxiredoxin 2 and glutathione peroxidase 3. Proteomics also revealed the decrease of apolipoproteins, and the presence of intracellular proteins in serum, which may indicate physical injury to tissues. Conclusions: Housing of gilts in individual stalls and diet change increase lipid and protein catabolism, oxidative stress, activate the innate immune system and cause a certain degree of tissue damage. We propose that valuable assays for stress assessment in gilts may be based on a score composed by a combination of salivary cortisol, lipid metabolites, innate immunity and oxidative stress markers and intracellular proteins.

Infection of dendritic cells with the American, but not the European, PRRSv genotype induces Treg (Foxp3+CD25+) cells

The Faseb Journal, Mar 1, 2008

Piglet nasal microbiota at weaning may influence the development of Glässer’s disease during the rearing period

BMC Genomics, 2016

Background: The microbiota, the ensemble of microorganisms on a particular body site, has been ex... more Background: The microbiota, the ensemble of microorganisms on a particular body site, has been extensively studied during the last few years, and demonstrated to influence the development of many diseases. However, these studies focused mainly on the human digestive system, while the populations in the respiratory tract have been poorly assessed, especially in pigs. The nasal mucosa of piglets is colonized by an array of bacteria, many of which are unknown. Among the early colonizers, Haemophilus parasuis also has clinical importance, since it is also the etiological agent of Glässer's disease. This disease produces economical losses in all the countries with pig production, and the factors influencing its development are not totally understood. Hence, the purpose of this work was to characterize the nasal microbiota composition of piglets, and its possible role in Glässer's disease development. Results: Seven farms from Spain (4 with Glässer's disease and 3 control farms without any respiratory disease) and three farms from UK (all control farms) were studied. Ten piglets from each farm were sampled at 3-4 weeks of age before weaning. The total DNA extracted from nasal swabs was used to amplify the 16S RNA gene for sequencing in Illumina MiSeq. Sequencing data was quality filtered and analyzed using QIIME software. The diversity of the nasal microbiota was low in comparison with other body sites, showing a maximum number of operational taxonomic units (OTUs) per pig of 1,603, clustered in five phyla. Significant differences were found at various taxonomical levels, when the microbiota was compared regarding the farm health status. Healthy status was associated to higher species richness and diversity, and UK farms demonstrated the highest diversity. Conclusions: The composition of the nasal microbiota of healthy piglets was uncovered and different phylotypes were shown to be significantly altered in animals depending on the clinical status of the farm of origin. Several OTUs at genus level were identified over-represented in piglets from control farms, indicating their potential as probiotics. Although we provide relevant data, fully metagenomic approaches could give light on the genes and metabolic pathways involved in the roles of the nasal microbiota to prevent respiratory diseases.

Marbofloxacin reaches high concentration in pig tonsils in a dose-dependent fashion: Marbofloxacin reaches high concentration in pig tonsils

J Vet Pharmacol Ther, 2010

Shedding of cephalosporin resistant Escherichia coli in pigs from conventional farms after early treatment with antimicrobials

Veterinary journal (London, England : 1997), Jan 4, 2016

This study assessed the dynamics of cephalosporin resistant (CR) E. coli populations during the l... more This study assessed the dynamics of cephalosporin resistant (CR) E. coli populations during the life cycle of pigs treated early in life with ceftiofur or tulathromycin. The study was conducted at eight conventional pig farms; four for each treatment with ceftiofur or tulathromycin. At each farm, 70 7-day-old piglets were divided into two groups: a control group (n = 30) and a treatment group (n = 40). Faecal samples were collected on day 0 and on days 2, 7 and 180 post-treatment. Sows were also sampled on day 0. CR E. coli were selected on MacConkey agar with ceftriaxone. On five farms, 7-day-old piglets excreted CR E. coli before treatment associated with the presence of CR E. coli in sows. The occurrence of CR E. coli positive animals decreased with increasing piglet age. The remaining three farms tested negative for CR E. coli during the study period. Results demonstrated great variability in the frequency of CR E. coli positive animals between farms, independent of treatment. T...