The effect of temperature on the boundary conditions of West Nile virus circulation in Europe.

West Nile virus (WNV) is a vector-borne flavivirus that causes an increasing number of human and equine West Nile fever cases in Europe. While the virus has been present in the Mediterranean basin and the Balkans since the 1960s, recent years have witnessed its northward expansion, with the first human cases reported in Germany in 2018 and the Netherlands in 2020. WNV transmission and amplification within mosquitoes are temperature-dependent. This study applies a mathematical modelling approach to assess the conditions under which WNV circulation occurs based on the proportion of mosquito bites on WNV-competent birds (dilution), vector-host ratios, mosquito season length and the observed daily temperature data. We modelled five distinct European regions where previous WNV circulation has been observed within the Netherlands, Germany, Spain, Italy, and Greece. We observed that the number of days in which the basic reproduction number (R0) is above one, increased over the last 40 years in all five regions. In the Netherlands, the number of days in which the R0 is above one, is 70% lower than in Spain. The temperature in Greece, Spain and Italy allowed for circulation under low vector-host ratios, and at a high dilution. On the other hand in the Netherlands and Germany, given the observed daily temperature, the thresholds for circulation requires a lower dilution and higher vector-host ratios. For the Netherlands, a short window of introductions between late May and mid-June would result in detectable outbreaks. Our findings revealed that the temperate maritime climate of the Netherlands allows WNV circulation primarily during warmer summers, and only under high vector-host ratios. This research contributes valuable insights into the dynamic relationship between temperature, vector properties, and WNV transmission, offering guidance for proactive strategies in addressing this emerging health threat in Europe.

A cross-sectional study on Campylobacter fetus subsp. venerealis prevalence and associated factors in Brazilian southern cattle farms.

Cattle farming is a major livestock activity with economic relevance in Rio Grande do Sul (RS), Brazil. However, this activity is still considered of intermediate to low technological level, and in this region, there are few epidemiologic reports of Campylobacter fetus subsp. venerealis (Cfv), the causative agent of bovine genital campylobacteriosis (BGC). Thus, we designed a cross-sectional study to assess the prevalence and Cfv-associated factors in cattle farms in RS, Brazil. In total, 99 farms were randomly selected to participate in the survey. Preputial mucus samples from selected bulls were collected twice (within a 15-day interval) and subjected to Cfv molecular detection. A farm was considered positive when at least one sample was positive for Cfv. Our findings indicate that the farm-level Cfv prevalence in RS is 67.67%. On average, the chance of a farm using natural service to be Cfv-positive increased approximately twice compared to farms that do not use natural service. We also determined that Cfv routine tests reduce the chance of a farm being positive by 92%. Therefore, both Cfv detection tests and the reduction of natural services decrease the chance of a farm being positive for Cfv. Finally, we conclude that Cfv is widely spread in Southern Brazil cattle farms and it is urgent the implementation of control measures to reduce Cfv prevalence in the target population.

In-silico characterization of the relationship between the Porcine reproductive and respiratory syndrome virus prevalence at the piglet and litter levels in a farrowing room.

BACKGROUND: Family oral fluids (FOF) sampling has been described as a sampling technique where a rope is exposed to sows and respective suckling litters and thereafter wrung to obtain fluids. PCR-based testing of FOF reveals presence of PRRS virus RNA only at the litter level, as opposed to conventional individual-animal-based sampling methods that demonstrate PRRSV RNA at the piglet level. The relationship between the PRRSV prevalence at the individual piglet level and at the litter level in a farrowing room has not been previously characterized. Using Monte Carlo simulations and data from a previous study, the relationship between the proportion of PRRSV-positive (viremic) pigs in the farrowing room, the proportion of litters in the farrowing room with at least one viremic pig, and the likely proportion of litters to be positive by a FOF RT-rtPCR test in a farrowing room was characterized, taking into account the spatial distribution (homogeneity) of viremic pigs within farrowing rooms. RESULTS: There was a linear relationship between piglet-level- and litter-level prevalence, where the latter was always larger than the former. When the piglet-level prevalence was 1%, 5%, 10%, 20%, and 50%, the true-litter level prevalence was 5.36%, 8.93%, 14.29%, 23.21%, and 53.57%, respectively. The corresponding apparent-litter prevalence by FOF was 2.06%, 6.48%, 11.25%, 21.60%, and 51.56%, respectively. CONCLUSION: This study provides matching prevalence estimates to help guide sample size calculations. It also provides a framework to estimate the likely proportion of viremic pigs, given the PRRSV RT-rtPCR positivity rate of FOF samples submitted from a farrowing room.

Main factors associated with foot-and-mouth disease virus infection during the 2001 FMD epidemic in Uruguay.

Large epidemics provide the opportunity to understand the epidemiology of diseases under the specific conditions of the affected population. Whilst foot-and-mouth disease (FMD) epidemics have been extensively studied in developed countries, epidemics in developing countries have been sparsely studied. Here we address this limitation by systematically studying the 2001 epidemic in Uruguay where a total of 2,057 farms were affected. The objective of this study was to identify the risk factors (RF) associated with infection and spread of the virus within the country. The epidemic was divided into four periods: (1) the high-risk period (HRP) which was the period between the FMD virus introduction and detection of the index case; (2) the local control measures period (LCM) which encompassed the first control measures implemented before mass vaccination was adopted; (3) the first mass vaccination, and (4) the second mass vaccination round. A stochastic model was developed to estimate the time of initial infection for each of the affected farms. Our analyses indicated that during the HRP around 242 farms were probably already infected. In this period, a higher probability of infection was associated with: (1) animal movements [OR: 1.57 (95% CI: 1.19-2.06)]; (2) farms that combined livestock with crop production [OR: 1.93 (95% CI: 1.43-2.60)]; (3) large and medium farms compared to small farms (this difference was dependent on regional herd density); (4) the geographical location. Keeping cattle only (vs farms that kept also sheep) was a significant RF during the subsequent epidemic period (LCM), and remained as RF, together with large farms, for the entire epidemic. We further explored the RF associated with FMDV infection in farms that raised cattle by fitting another model to a data subset. We found that dairy farms had a higher probability of FMDV infection than beef farms during the HRP [OR: 1.81 (95% CI: 1.12-2.83)], and remained as RF until the end of the first round of vaccination. The delay in the detection of the index case associated with unrestricted animal movements during the HRP may have contributed to this large epidemic. This study contributes to the knowledge of FMD epidemiology in extensive production systems.

Cystoisospora suis infection in suckling piglets in Brazil: Prevalence and associated factors.

Piglet coccidiosis is a parasitic disease caused by the protozoan Cystoisospora suis, which is regarded as the most prevalent gastrointestinal parasite in intensive pig farms. Despite the availability of highly effective chemo-metaphylaxis (toltrazuril), coccidiosis is still prevalent in European and other countries. We conducted a cross-sectional study on swine farms in Brazil, to assess the prevalence of Cystoisospora suis in fecal samples and determined the associated factors. In total, 666 litters from 50 farms were sampled twice within one week between samplings (mean age at sampling: 10.75 and 17.7 days). Of 666 litters, 225 (33.8%) were positive at least once, and the expected within farm prevalence of C. suis oocysts was 32.9% (25.4-41.3%; 95% confidence interval). Oocysts were more prevalent in fecal samples collected from farms with diarrhea (odds ratio = 6.75). The room temperature was also positively associated with oocyst detection; a one-degree increase in room temperature increased the chance of a litter being positive by 23.2%. Up-to-date, this is the most comprehensive technical evidence of factors associated with C. suis infection in Brazilian industrial piglet farms.

A Systematic Review and Meta-Analysis on the Occurrence of Toxoplasmosis in Animals Slaughtered in Brazilian Abattoirs.

Toxoplasmosis is mainly transmitted to human beings through the consumption of contaminated food, and several outbreaks caused by infected meat consumption have been reported in Brazil. We performed a systematic literature review on the prevalence and risk factors for toxoplasmosis in slaughtered animals and performed a meta-analysis of its prevalence for different species and regions. Furthermore, we also discussed the infectivity of seropositive animals, risk factors, and preventive strategies. In the meta-analysis, the overall prevalence estimates for poultry, ostrich, goats, swine, equids, sheep, and bovines were 42.4, 40.4, 23.0, 19.9, 19.1, 17.3, and 16.2%, respectively. Regarding the Brazilian regions, the highest prevalence values were detected for bovines and equids in the South (32.3 and 34.4%, respectively) and swine, goats, sheep, and poultry in the Northeast (29.3, 23.0, 22.9, and 69.8%%, respectively). High proportions of Toxoplasma gondii viability in bioassay conducted on seropositive animals were seen for sheep (34/40 = 85%) and swine (12/15 = 80%). Toxoplasma gondii infections are widespread on Brazilian farms, and the summarized data allow the establishment of high-priority areas and/or species for the adoption of preventive strategies to control this parasite at different levels of the food chain.

Critically Important Antimicrobial Resistance Trends in Salmonella Derby and Salmonella Typhimurium Isolated from the Pork Production Chain in Brazil: A 16-Year Period.

Knowledge about antimicrobial resistance in Salmonella is relevant due to its importance in foodborne diseases. We gathered data obtained over 16 years in the southern Brazilian swine production chain to evaluate the temporal evolution of halo for carbapenem, and the MIC for third-generation cephalosporins, fluoroquinolone, and polymyxin in 278 Salmonella Derby and Typhimurium isolates. All antimicrobial resistance assays were performed in accordance with EUCAST. To assess the diameter halo, we used a mixed linear model, and to assess the MIC, an accelerated failure time model for interval-censored data using an exponential distribution was used. The linear predictor of the models comprised fixed effects for matrix, serovar, and the interaction between year, serovar, and matrix. The observed halo diameter has decreased for ertapenem, regardless of serovars and matrices, and for the serovar Typhimurium it has decreased for three carbapenems. The MIC for ciprofloxacin and cefotaxime increased over 16 years for Typhimurium, and for Derby (food) it decreased. We did not find evidence that the MIC for colistin, ceftazidime, ciprofloxacin (Derby), or cefotaxime (food Typhimurium and animal Derby) has changed over time. This work gave an overview of antimicrobial resistance evolution from an epidemiological point of view and observed that using this approach can increase the sensitivity and timeliness of antimicrobial resistance surveillance.

Longitudinal study on the prevalence of extended spectrum cephalosporins-resistant Escherichia coli colonization in Dutch veal farms.

A longitudinal study was performed to investigate the prevalence of Extended-Spectrum Cephalosporin-Resistant (ESC-R) Escherichia coli colonization in Dutch veal farms. Rectal swabs from 683 calves born in 13 Dutch dairy farms were collected one day prior to transportation to the veal farm at 14 or 28 days of age, and at 5 different time points 8 Dutch veal farms. In addition, characteristics of the calf, cows, and farm management were collected. Rectal swabs were selectively cultured for ESC-R E. coli. In total, 1202 ESC-R E. coli isolates were recovered. Overall, the prevalence of ESC-R E. coli increased from 24.4 % at one day prior to transportation to 57.3 % in week two after arrival of calves at the veal farm. No associations were found between the presence of ESC-R E. coli at the dairy or veal farm and age of transportation, sex and breed. The presence of ESC-R E. coli in week 6, 10, and 18 at the veal farm was positively associated with the presence of ESC-R E. coli in week 10, 18, and 24, respectively (p < 0.05). Individual antibiotic treatments applied before week 2 and 6 upon arrival to the veal farms tended to increase the ESC-R E. coli colonization frequency. Our results indicate that ESC-R E. coli colonization frequency substantially increases after arrival of calves on the veal farm. In addition to individual antibiotic treatments, it is considered likely that frequently applied batch antibiotic treatments are also implicated in the ESC-R E. coli colonization frequency.

Effect of pooling family oral fluids on the probability of PRRSV RNA detection by RT-rtPCR.

Family oral fluids (FOFs) are an aggregate sample type shown to be a cost-efficient and convenient option for determining the porcine reproductive and respiratory syndrome virus (PRRSV) status of weaning age pigs. This study investigates the effect of pooling PRRSV-positive FOF samples with PRRSV-negative FOF samples at different levels (1/3, 1/5, 1/10, 1/20) on the probability of PRRSV RNA detection by reverse-transcription real-time polymerase chain reaction (RT-rtPCR). Mathematical models were built to assess how much the probability of RT-rtPCR PRRSV detection changed with increasing proportion of PRRSV-positive samples present within pools and how partially sampling a farrowing room influenced the probability of RT-rtPCR detection of PRRSV RNA in pooled samples at different prevalence scenarios. A general example of a guideline for FOF-based sampling under different prevalence scenarios to detect PRRSV RNA by RT-rtPCR with at least 95 % certainty is presented. At the sample level, the probability of detecting PRRSV RNA by RT-rtPCR decreased from 100 % to 87 %, 68 %, and 26 % when diluting up to 1/20 for PRRSV positive FOF having an initial Cycle threshold (Ct) below 34, between 34 and 36, or above 36, respectively. When PRRSV prevalence is near-zero (1 or 2 litters positive out of 56), the most cost-efficient farrowing room sampling strategy to detect PRRSV RNA with at least 95 % certainty was pooling FOF samples up to 1/10; at higher prevalence (≥ 3 of 56 litters positive), the most cost-efficient strategy was submitting samples in pools of 20. Subsampling a farrowing room for FOF pools was also demonstrated to be a valuable cost-saving strategy. Overall, based on the conditions of this study, pooling FOFs up to 1/20 is a valid option in situations of cost constraint and regardless of pooling level chosen, capturing as many litters as possible improves the probability of PRRSV detection.

Antimicrobial resistance in commensal Escherichia coli and Enterococcus spp. isolated from pigs subjected to different antimicrobial administration protocols.

The antimicrobial resistance (AMR) in human and animal pathogens is a global concern, and antimicrobial use (AMU) is considered the most important driver for its increase. The aim of this study was to assess AMR in Escherichia coli and Enterococcus spp. in faecal samples of pigs subjected to four different AMU protocols from birth to finishing: G1, no in-feed antimicrobials; G2: a total average dose 6018 mg antimicrobials/pig; G3: a total average dose 8127 mg antimicrobials/pig; and G4: a total average dose 15,678 mg antimicrobials/pig. Faecal samples were collected at six time points and AMR was assessed in both bacteria. The microbiota composition was assessed by 16S rRNA sequencing. Minor differences on the microbiota profile was observed among groups, but a lower Firmicutes:Bacteroidetes ratio was noted in G4. Escherichia coli and Enterococcus spp. strains isolated from all groups showed a high level of multi-drug resistance (MDR). The amount of antimicrobials used was significantly positively associated with the probability of MDR in both bacteria. Approximately 43% of the variation in MIC90 for colistin could be explained by AMU, and a one-day increase in administration of colistin increased MIC90 by 0.05 µg mL-1. In conclusion, the results suggest that the higher the use of antimicrobials in farms, the higher the MDR frequency and resistance to the highest priority critically important antimicrobials for humans in commensal gut bacteria of pigs.

Impact of changes of horse movement regulations on the risks of equine infectious anemia: A risk assessment approach.

Equine infectious anemia virus (EIAV) is a transboundary disease affecting a large number of equines worldwide. In this study, we assessed the transmission risk of EIAV in Rio Grande do Sul, Brazil. Serum samples from 1010 animals from 341 farms were initially analyzed using agar gel immunodiffusion to detect viral antibodies, and no antibody-positive animals were found. A risk assessment stochastic model was applied to generate the expected number of potential infections per month and to estimate the time to new infections. Our results estimated 6.5 months as the interval for new infections in the worst-case scenario. Among the variables evaluated, the number of transported animals and the test sensitivity influenced the model the most. These results were then used to revisit the impact of EIAV control regulations, which triggered a change in the diagnostic testing required for animal movement, in which the validity of a negative test for EIAV was extended from 60 to 180 days. Finally, revisiting the annual average of infected farms before and after the new regulation, the number of infected farms increased pre-implementation, and then, the number of culled animals increased, which should impact future EIAV incidence in this region. Our results demonstrated the importance of constant reviews of disease control programs and provided quantitative-based knowledge for decision-makers in official veterinary services.

Biosecurity practices associated with influenza A virus seroprevalence in sows from southern Brazilian breeding herds.

Influenza A virus (IAV) infection is a recognized cause of acute respiratory disease in pigs that can culminate in the decline of performance due to increasing feed conversion and costs of antimicrobial drugs to control secondary infections. Biosecurity practices are the key to prevent transmission of highly contagious agents. The aim of this study was to assess the effect of biosecurity practices on IAV seroprevalence through a cross-sectional study carried out in 404 sows from 21 herds. An indirect ELISA was used to detect antibodies against a nucleoprotein of IAV. To evaluate IAV subtypes (H1N1pdm09, H1N2 and H3N2), all samples positive by ELISA were tested using the hemagglutination inhibition assay (HI). Prevalence ratios (PR) estimates were calculated using multivariate Poisson regression accounted with survey weights. Sixty-four percent (261/404) of sows were positive in the rNP-ELISA and the estimated prevalence was 63.9% (95% CI 55%-73%). All farms had at least one seropositive sow; the frequency of IAV subtypes found in seropositive sows was 51.9% for H1N1pdm09, 38.1% for codetection H1N1pdm09 and H1N2, 8.6% for H1N2, and 0.6% for codetection H1N1pdm09 and H3N2, and 19 herds presented coinfection of H1N1 pdm09 and H1N2. Variables significantly associated with IAV seroprevalence found in the final model were 'bird-proof net' (PR = 0.75; 95% CI: 0.65-0.86) and 'gilt acclimatization unit' (PR = 0.57, 95% CI: 0.50-0.66), showing a protective effect against IAV seroprevalence, and 'external replacement', which had a positive effect on IAV seroprevalence (PR = 1.38, 95% CI: 1.17-1.64). This study suggests that preventing contact among wild species and swine and using an adaptation area for animals before entry into the herd can be strategies to control the influenza virus in breeding herds.

A Stochastic Model to Assess the Effect of Meat Inspection Practices on the Contamination of the Pig Carcasses.

The objective of meat inspection is to promote animal and public health by preventing, detecting, and controlling hazards originating from animals. With the improvements of sanitary level in pig herds, the hazards profile has shifted and the inspection procedures no longer target major foodborne pathogens (i.e., not risk based). Additionally, carcass manipulations performed when searching for macroscopic lesions can lead to cross-contamination. We therefore developed a stochastic model to quantitatively describe cross-contamination when consecutive carcasses are submitted to classic inspection procedures. The microbial hazard used to illustrate the model was Salmonella, the data set was obtained from Brazilian slaughterhouses, and some simplifying assumptions were made. The model predicted that due to cross-contamination during inspection, the prevalence of contaminated carcass surfaces increased from 1.2% to 95.7%, whereas the mean contamination on contaminated surfaces decreased from 1 logCFU/cm² to -0.87 logCFU/cm², and the standard deviations decreased from 0.65 to 0.19. These results are explained by the fact that, due to carcass manipulations with hands, knives, and hooks, including the cutting of contaminated lymph nodes, Salmonella is transferred to previously uncontaminated carcasses, but in small quantities. These small quantities can easily go undetected during sampling. Sensitivity analyses gave insight into the model performance and showed that the touching and cutting of lymph nodes during inspection can be an important source of carcass contamination. The model can serve as a tool to support discussions on the modernization of pig carcass inspection.

Extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli isolates collected from diseased food-producing animals in the GERM-Vet monitoring program 2008-2014.

The aim of this study was to identify extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli collected from diseased food-producing animals in Germany. A total of 6849 E. coli isolates, collected from diseased cattle, pigs and poultry in the German national monitoring program GERM-Vet (2008-2014), were characterized by antimicrobial susceptibility testing and screened for the ESBL phenotype. ESBL genes were identified by PCR and sequencing. The isolates were further characterized by PCR-based phylotyping. The 419/6849 (6.1%) ESBL-producers identified included 324/2896 (11.2%) isolates from cattle, 75/1562 (4.8%) from pigs and 20/2391 (0.8%) from poultry. The ESBL genes detected were: blaCTX-M-1 (69.9%), blaCTX-M-15 (13.6%), blaCTX-M-14 (11.7%), blaTEM-52 (1.9%), blaSHV-12 (1.4%), blaCTX-M-3 (1.0%), and blaCTX-M-2 (0.5%). The phylogroup A was the dominant phylogroup (57.0%) followed by phylogroups D (23.4%), B1 (17.9%), and B2 (1.7%). Bovine isolates belonged predominantly to the phylogroups A and D, whereas the porcine and avian isolates mainly belonged to A and B1. The majority of the ESBL-producing isolates found in each phylogroup were from animals suffering from gastrointestinal infections. In 399/419 isolates (95.2%), additional resistance to non-ß-lactam antibiotics was seen. Multidrug-resistance [resistance to aminoglycosides, fluoro(quinolones), sulphonamides, tetracyclines, and trimethoprim] was seen in 369/419 (88.1%) isolates, which may facilitate the co-selection of ESBL genes, when located on the same mobile genetic element as the others resistance genes, and may compromise the therapeutic options.

Effect of slaughterhouse and day of sample on the probability of a pig carcass being Salmonella-positive according to the Enterobacteriaceae count in the largest Brazilian pork production region.

Sources of contamination of carcasses during slaughter include infected pigs as well as environmentally related sources. There are many microbial indicators that can be used in the processing of food to assess food hygiene and the safety of food processing. The presence of some microbial indicators can be viewed as a result of direct or indirect contamination of a food with fecal material. The presence of Enterobacteriaceae is often used as a hygiene indicator, as they are found both in the environment and in the intestine of warm-blooded animals. An association between Salmonella isolation and Enterobacteriaceae count (EC) on pre-chill carcasses has been described, however the impact of slaughterhouse and the day of sampling on the occurrence of Salmonella has not been previously investigated. To this end, mixed logistic regressions (MLRs) with random effects and fixed slopes were performed to assess the change in EC and its correlation with Salmonella occurrence using two data sets. The first describes the EC and Salmonella isolation in 60 pork carcasses in one slaughterhouse sampled at 11 different slaughter steps, including the carcass as a random effect. The second describes the EC and Salmonella isolation on 1150 pre-chill carcasses sampled in 13 slaughterhouses over 230 sampling days, and the model combined two random intercepts, slaughterhouse and date of sampling nested with slaughterhouse (day/slaughterhouse). Statistically significant associations (p<0.0001) between the log of the EC and Salmonella occurrence were found in all models. Nevertheless, although a strong association was found between Enterobacteriaceae and Salmonella contamination in pork carcasses, this association was not constant, given that there was a high variation in the probability of a carcass being positive for Salmonella according to the EC mainly between days of samples. The effect of the day of sampling on Salmonella prevalence was so large that the predictive value of the EC count for Salmonella isolation on a daily basis was compromised. It is possible that on some days batches with a high prevalence of Salmonella carriers shedding a high number of Salmonella were slaughtered. On these days, the potential for contamination/cross-contamination of carcasses will be so large that even hygienic slaughter, confirmed by the low EC on carcasses, will not be able to prevent the presence of Salmonella on some carcasses. The results of this study demonstrate that, despite the statistically significant association found, it may be difficult to predict when hygiene failure measured via EC actually indicates Salmonella contamination, and neither the inverse.