Metagenomic comparison of the faecal and environmental resistome on Irish commercial pig farms with and without zinc oxide and antimicrobial usage.

BACKGROUND: Antimicrobials and heavy metals such as zinc oxide (ZnO) have been commonly used on Irish commercial pig farms for a 2-week period post-weaning to help prevent infection. In 2022, the prophylactic use of antimicrobials and ZnO was banned within the European Union due to concerns associated with the emergence of antimicrobial resistance (AMR) and contamination of the environment with heavy metals. In this study, faecal and environmental samples were taken from piglets during the weaning period from ten commercial farms, of which five farms used antimicrobial or ZnO prophylaxis (AB-ZnO farms) and five which had not used antimicrobials or ZnO for the previous 3 years (AB-ZnO free farms). A total of 50 samples were compared using a metagenomic approach. RESULTS: The results of this study showed some significant differences between AB-ZnO and AB-ZnO free farms and suggested positive selection for AMR under antimicrobial and ZnO treatment. Moreover, strong differences between environmental and faecal samples on farms were observed, suggesting that the microbiome and its associated mobile genetic elements may play a key role in the composition of the resistome. Additionally, the age of piglets affected the resistome composition, potentially associated with changes in the microbiome post-weaning. CONCLUSIONS: Overall, our study showed few differences in the resistome of the pig and its environment when comparing AB-ZnO farms with AB-ZnO free farms. These results suggest that although 3 years of removal of in-feed antimicrobial and ZnO may allow a reduction of AMR prevalence on AB-ZnO farms, more time, repeated sampling and a greater understanding of factors impacting AMR prevalence will be required to ensure significant and persistent change in on-farm AMR.

Antimicrobial Resistance in Commensal Escherichia coli of the Porcine Gastrointestinal Tract.

Antimicrobial resistance (AMR) in Escherichia coli of animal origin presents a threat to human health. Although animals are not the primary source of human infections, humans may be exposed to AMR E. coli of animal origin and their AMR genes through the food chain, direct contact with animals, and via the environment. For this reason, AMR in E. coli from food producing animals is included in most national and international AMR monitoring programmes and is the subject of a large body of research. As pig farming is one of the largest livestock sectors and the one with the highest antimicrobial use, there is considerable interest in the epidemiology of AMR in E. coli of porcine origin. This literature review presents an overview and appraisal of current knowledge of AMR in commensal E. coli of the porcine gastrointestinal tract with a focus on its evolution during the pig lifecycle and the relationship with antimicrobial use. It also presents an overview of the epidemiology of resistance to extended spectrum cephalosporins, fluoroquinolones, and colistin in pig production. The review highlights the widespread nature of AMR in the porcine commensal E. coli population, especially to the most-used classes in pig farming and discusses the complex interplay between age and antimicrobial use during the pig lifecycle.

Special Issue-Resistant Staphylococci in Animals.

Staphylococci figure prominently among those bacteria demonstrating antimicrobial resistance (AMR) and are thus responsible for significant problems concerning the treatment of the animals and humans that they infect [...].

Improving infection control in a veterinary hospital: a detailed study on patterns of faecal contamination to inform changes in practice.

BACKGROUND: The main purpose of this study was to investigate the cleanliness and microbial burden of a veterinary hospital to establish the extent of cross-contamination with faecal bacteria as an aid to reducing nosocomial infections. Enterococci and Escherichia coli were used as faecal indicator organisms as they can survive on inanimate surfaces for months and pose a threat to animal health. The study consisted of several elements: (i) a cross-sectional study to identify sites currently contaminated with faecal organisms that could be usefully included in a longitudinal study, (ii) a 3-week longitudinal study to identify sites from which faecal bacteria were repeatedly recovered, (iii) once-off monitoring of hand hygiene, (iv) a review of all hospitalised cases with confirmed E. coli or enterococcal infection during the 8-week study period to investigate possible hospital-acquired (HAI) infection and relationship with environmental contamination. Environmental surface and hand hygiene were assessed using 3M™ Clean-Trace™ ATP test, 3M™ Petrifilm™ plates and bacteriological culture of Enterococcus species and E. coli. Cross contamination was assessed using results of antimicrobial susceptibility testing. RESULTS: In the cross-sectional study, 26 of 113 (24.5%) of sites sampled exceeded the accepted microbial threshold (2.5 CFU/cm2) and Enterococcus species were isolated from 31 (27.4%) and E. coli from 9 (7.9%) of 113 samples. Organic residue and microbial levels were high in the dog kennels even after cleaning and faecal organisms were also recovered from sites such as the dispensary, a student computer and staff common room. Four of 51 (7.8%) hand samples were contaminated with faecal bacteria. Nine sites were monitored on three occasions in the longitudinal study and a total of 23 Escherichia coli and 6 Enterococcus species were recovered. Seven of the nine sites were positive for faecal organisms on more than one occasion. There was no change in cleanliness or microbial burden over 3 weeks. Twenty-one of the 73 isolates (28.8%) recovered during all parts of the study were multi-drug resistant. Enterococci and E. coli isolates with similar resistance patterns were recovered from the environment in the large and small animal hospitals and from a small number of patients during the same timeframe, suggesting possible hospital acquired infections. CONCLUSIONS: Results suggested that movement between the small and large animal hospital areas may have been responsible for cross-contamination and possible hospital-acquired infections. The data show that cross-sectional and longitudinal monitoring of faecal contamination across all hospital areas can play an important role in informing review of infection control protocols in veterinary hospital settings. Changes in practices in the hospital based on results generated are outlined.

Insights on the effects of antimicrobial and heavy metal usage on the antimicrobial resistance profiles of pigs based on culture-independent studies.

Antimicrobial resistance is a global threat to human, animal, and environmental health. In pig production, antimicrobials and heavy metals such as zinc oxide are commonly used for treatment and prevention of disease. Nevertheless, the effects of antimicrobials and heavy metals on the porcine resistome composition and the factors influencing this resistance profile are not fully understood. Advances in technologies to determine the presence of antimicrobial resistance genes in diverse sample types have enabled a more complete understanding of the resistome and the factors which influence its composition. The aim of this review is to provide a greater understanding of the influence of antimicrobial and heavy metal usage on the development and transmission of antimicrobial resistance on pig farms. Furthermore, this review aims to identify additional factors that can affect the porcine resistome. Relevant literature that used high-throughput sequencing or quantitative PCR methods to examine links between antimicrobial resistance and antimicrobial and heavy metal use was identified using a systematic approach with PubMed (NCBI), Scopus (Elsevier), and Web of Science (Clarivate Analytics) databases. In total, 247 unique records were found and 28 publications were identified as eligible for inclusion in this review. Based on these, there is clear evidence that antimicrobial and heavy metal use are positively linked with antimicrobial resistance in pigs. Moreover, associations of genes conferring antimicrobial resistance with mobile genetic elements, the microbiome, and the virome were reported, which were further influenced by the host, the environment, or the treatment itself.

Comparison of antimicrobial resistant Escherichia coli isolated from Irish commercial pig farms with and without zinc oxide and antimicrobial usage.

BACKGROUND: The prophylactic use of antimicrobials and zinc oxide (ZnO) in pig production was prohibited by the European Union in 2022 due to potential associations between antimicrobial and heavy metal usage with antimicrobial resistance (AMR) and concerns regarding environmental pollution. However, the effects of their usage on the bacterial AMR profiles on commercial pig farms are still not fully understood and previous studies examining the effect of ZnO have reported contrasting findings. The objective of this study was to examine the effects of antimicrobial and ZnO usage on AMR on commercial pig farms. Faecal and environmental samples were taken on 10 Irish commercial farms, of which 5 farms regularly used ZnO and antimicrobials (amoxicillin or sulphadiazine-trimethoprim) for the prevention of disease. The other 5 farms did not use ZnO or any other form of prophylaxis. Escherichia coli numbers were quantified from all samples using non-supplemented and supplemented Tryptone Bile X-glucuronide agar. RESULTS: In total 351 isolates were phenotypically analysed, and the genomes of 44 AmpC/ESBL-producing E. coli isolates from 4 farms were characterised using whole-genome sequencing. Phenotypic analysis suggested higher numbers of multi-drug resistant (MDR) E. coli isolates on farms using prophylaxis. Furthermore, farms using prophylaxis were associated with higher numbers of isolates resistant to apramycin, trimethoprim, tetracycline, streptomycin, and chloramphenicol, while resistance to ciprofloxacin was more associated with farms not using any prophylaxis. Thirty-four of the 44 AmpC/ESBL-producing E. coli strains harboured the blaCTX-M-1 resistance gene and were multi drug resistant (MDR). Moreover, network analysis of plasmids and analysis of integrons showed that antimicrobial and biocide resistance genes were frequently co-located on mobile genetic elements, indicating the possibility for co-selection during antimicrobial or biocide usage as a contributor to AMR occurrence and persistence on farms. CONCLUSIONS: The results of this study showed evidence that antimicrobial and ZnO treatment of pigs post-weaning can favour the selection and development of AMR and MDR E. coli. Co-location of resistance genes on mobile genetic elements was observed. This study demonstrated the usefulness of phenotypic and genotypic detection of antimicrobial resistance by combining sequencing and microbiological methods.

Examining the impact of zinc on horizontal gene transfer in Enterobacterales.

Antimicrobial resistance is one of the main international health concerns for humans, animals, and the environment, and substantial efforts have focused on reducing its development and spread. While there is evidence for correlations between antimicrobial usage and antimicrobial resistance development, specific information on the effect of heavy metal/antimicrobial usage on bacterial conjugation is more limited. The aim of this study was to investigate the effects of zinc and antimicrobials in different concentrations on horizontal gene transfer of an ampicillin resistance gene, using a multi-drug resistant Escherichia coli donor strain and three different Salmonella enterica serovars as recipient strains. Differences in conjugation frequencies for the different Salmonella recipients were observed, independent of the presence of zinc or the antimicrobials. Selective pressure on the recipient strains, in the form of ampicillin, resulted in a decrease in conjugation frequencies, while, the presence of rifampicin resulted in increases. Zinc exposure affected conjugation frequencies of only one of the three recipient strains, thus the effect of zinc on conjugation frequencies seemed to be concentration and strain dependent. Furthermore, differences in growth rates due to plasmid carriage were observed for one of the Salmonella strains.

A survey of food-borne and antimicrobial resistance-harbouring bacteria in meat by-products from knackeries and associated equipment and kennels.

BACKGROUND: In Ireland, meat by-products (MBP) harvested at knackeries from farmed animals that have not died of an infectious or systemic disease are legally permitted to be fed to dogs in kennels and packs of hounds. There is limited information available on the risks of spreading foodborne bacteria or antimicrobial resistant (AMR) determinants to dogs, their handlers or the associated environment. The aim of this study was to investigate the distribution of Salmonella serovars, Listeria monocytogenes, Campylobacter species, enterococci, their associated AMR determinants and the level of Escherichia coli in samples of MBP from knackeries and associated equipment and kennels. For this purpose, 313 fresh and 208 frozen MBP samples from 22 knackeries, 16 swabs of mincing equipment from two of the knackeries and 138 swabs from kennels adjacent to seven of the knackeries were collected and processed over a 12-month period. RESULTS: From the 521 MBP samples analysed, a total of 77 Salmonella (14.8%), 101 L. monocytogenes (19.4%), 12 Campylobacter (2.3%), 271 Enterococcus faecalis (52.0%) and 127 Enterococcus faecium (24.4%) strains were recovered. The 154 analysed environmental samples from kennels and mincing equipment yielded 194 isolates (3 Salmonella, 85 E. coli, 76 E. faecalis and 30 E. faecium.). E. coli was quantifiable in 423 of the 521 MBP samples with log counts per gram ranging between 1 and 6. AMR characterisation of 168 E. coli, enterococci and Salmonella isolates from MBP and environmental samples showed high levels of AMR including multi-drug resistance (MDR) with 63.6%, 9.1%, 29% and 45.8% of E. coli, Salmonella, E. faecalis and E. faecium isolates, respectively showing resistance to three or more antimicrobials (MDR) CONCLUSIONS: The findings of this survey confirm that MBP from fallen animals contain high levels of zoonotic and AMR-harbouring bacteria that pose a risk of transmission to dogs, their handlers, and the environment.

Antimicrobial resistance in Escherichia coli isolated from on-farm and conventional hatching broiler farms in Ireland.

BACKGROUND: On-farm hatching (OH) systems are becoming more common in broiler production. Hatching conditions differ from conventional farms as OH chicks avoid exposure to handling, transport, post-hatch water and feed deprivation. In contrast, chicks in conventional hatching conditions (CH) are exposed to standard hatchery procedures and transported post hatching. The objectives of this pilot study were to investigate the prevalence and frequency of Escherichia coli resistant to antimicrobials, including presumptive ESBL/AmpC-producing E. coli, isolated from environmental and faecal samples from OH versus CH hatching systems, and to investigate the presence of ESBL/AmpC-producing encoding genes. RESULTS: Environmental samples were collected from one flock in 10 poultry farms (5 OH farms, 5 CH farms) on day 0 post disinfection of the facilities to assess hygiene standards. On D10 and D21 post egg/chick arrival onto the farm, samples of faeces, boot swabs and water drinker lines were collected. E. coli were isolated on MacConkey agar (MC) and MacConkey supplemented with cefotaxime (MC+). Few E. coli were detected on D0. However, on D10 and D21 E. coli isolates were recovered from faeces and boot swabs. Water samples had minimal contamination. In this study, 100% of cefotaxime resistant E. coli isolates (n=33) detected on selective media and 44% of E. coli isolates (84/192) detected on nonselective media were multidrug resistant (MDR). The antimicrobial resistance (AMR) genotype for the 15 ESBL/AmpC producing isolates was determined using multiplex PCR. Six of these were selected for Sanger sequencing of which two were positive for blaCMY-2, two for blaTEM-1 and two were positive for both genes. CONCLUSIONS: There was no difference in E. coli isolation rates or prevalence of AMR found between the OH versus CH systems, suggesting that the OH system may not be an additional risk of resistant E. coli dissemination to broilers compared to the CH systems. The frequency of ß-lactam resistant E. coli in boot swab and faeces samples across both OH (24/33 (73%)) and CH (9/33 (27%)) systems may indicate that hatcheries could be a reservoir and major contributor to the transmission of AMR bacteria to flocks after entry to the rearing farms.

Prevalence of Salmonella spp. in slaughter pigs and carcasses in Irish abattoirs and their antimicrobial resistance.

BACKGROUND: Salmonella is an important zoonotic pathogen and is one of the main causes of foodborne outbreaks and infections in the European Union. Pigs are a significant reservoir and are frequently subclinical carriers of this organism. Salmonella can be shed in the faeces allowing infection to spread to other pigs, the environment, transport vehicles, lairages and other areas. Inadvertent spillage of gut contents during the slaughter process also leads to contamination. A pig Salmonella control programme has operated in Ireland since 2002 but many local surveys and an EUMS baseline survey in 2008 continued to indicate high levels of the organism in the pig sector. The objectives of this study were to generate updated information on the prevalence of Salmonella spp, in slaughter pigs and carcasses in Irish abattoirs. Five pigs from each of 164 herds were randomly sampled over a 14-week period during 2016. One sample from each of the five pigs of; caecal content, ileo-caecal lymph nodes and carcass swabs (pre-chill) were collected. The five caeca and lymph node samples from each herd were processed as one pool of caecal samples and one pool of lymph node samples, respectively, while the five carcass swabs were tested as individual samples. All isolates were characterised by serotyping and antimicrobial susceptibility. RESULTS: In total, 235 Salmonella spp. were isolated from 820 individual carcass swabs, 164 pooled lymph nodes and 164 caecal contents. Salmonella spp. were isolated from 54.3% of the caecal contents and from 31.7% of the ileo-caecal lymph node sample pools. A total of 11.5% of carcass-swab samples yielded Salmonella spp. S. Typhimurium 4,[5],12:i:1,2 or its monophasic variant 4,[5],12:i:-: predominated among isolates from all positive samples; accounting for 73% of lymph nodes, 68% of caecal contents and 56% of carcass swab isolates. S. London and S. Derby were the next most common isolated serotypes. CONCLUSIONS: These results confirm continuing high levels of Salmonella in fattening pigs in Ireland although reductions in carcass contamination compared to previous surveys were noted. A high prevalence of Salmonella in lymph nodes suggests that it remains a significant problem pre slaughter and a challenge to abattoirs in adhering to process hygiene requirements. The high prevalence of monophasic S. Typhimurim 4,[5],12:i:-: is of serious concern. Therefore, it is important to identify contributing factors in the dissemination of this pathogen in the pork industry in order to minimise the risk of human salmonellosis cases.

Bovine milk somatic cell transcriptomic response to Staphylococcus aureus is dependent on strain genotype.

BACKGROUND: Mastitis is an economically important disease of dairy cows with Staphylococcus aureus a major cause worldwide. Challenge of Holstein-Friesian cows demonstrated that S. aureus strain MOK124, which belongs to Clonal Complex (CC)151, caused clinical mastitis, while strain MOK023, belonging to CC97, caused mild or subclinical mastitis. The aim of this study was to elucidate the molecular mechanisms of the host immune response utilising a transcriptomic approach. Milk somatic cells were collected from cows infected with either S. aureus MOK023 or MOK124 at 0, 24, 48, 72 and 168 h post-infection (hpi) and analysed for differentially expressed (DE) genes in response to each strain. RESULTS: In response to MOK023, 1278, 2278, 1986 and 1750 DE genes were found at 24, 48, 72 and 168 hpi, respectively, while 2293, 1979, 1428 and 1544 DE genes were found in response to MOK124 at those time points. Genes involved in milk production (CSN1, CSN10, CSN1S2, CSN2, a-LACTA and PRLR) were downregulated in response to both strains, with a more pronounced decrease in the MOK124 group. Immune response pathways such as NF-κB and TNF signalling were overrepresented in response to both strains at 24 hpi. These immune pathways continued to be overrepresented in the MOK023 group at 48 and 72 hpi, while the Hippo signalling, extracellular matrix interaction (ECM) and tight junction pathways were overrepresented in the MOK124 group between 48 and 168 hpi. Cellular composition analysis demonstrated that a neutrophil response was predominant in response to MOK124, while M1 macrophages were the main milk cell type post-infection in the MOK023 group. CONCLUSIONS: A switch from immune response pathways to pathways involved in maintaining the integrity of the epithelial cell layer was observed in the MOK124 group from 48 hpi, which coincided with the occurrence of clinical signs in the infected animals. The higher proportion of M1 macrophages in the MOK023 group and lack of substantial neutrophil recruitment in response to MOK023 may indicate immune evasion by this strain. The results of this study highlight that the somatic cell transcriptomic response to S. aureus is dependent on the genotype of the infecting strain.

Risk Factors for Antimicrobial Use on Irish Pig Farms.

The threat to public health posed by antimicrobial resistance in livestock production means that the pig sector is a particular focus for efforts to reduce antimicrobial use (AMU). This study sought to investigate the risk factors for AMU in Irish pig production. Antimicrobial use data were collected from 52 farrow-to-finish farms. The risk factors investigated were farm characteristics and performance, biosecurity practices, prevalence of pluck lesions at slaughter and serological status for four common respiratory pathogens and vaccination and prophylactic AMU practices. Linear regression models were used for quantitative AMU analysis and risk factors for specific AMU practices were investigated using logistic regression. Farms that milled their own feed had lower total AMU (p < 0.001), whereas higher finisher mortality (p = 0.043) and vaccinating for swine influenza (p < 0.001) increased AMU. Farms with higher prevalence of pericarditis (p = 0.037) and lung abscesses (p = 0.046) used more group treatments. Farms with higher prevalence of liver milk spot lesions (p = 0.018) and farms practising prophylactic AMU in piglets (p = 0.03) had higher numbers of individual treatments. Farms practising prophylactic AMU in piglets (p = 0.002) or sows (p = 0.062) had higher use of cephalosporins and fluoroquinolones. This study identified prophylactic use and respiratory disease as the main drivers for AMU in Irish pig production. These findings highlight areas of farm management where interventions may aid in reducing AMU on Irish pig farms.

Detection of bla OXA-1, bla TEM-1, and Virulence Factors in E. coli Isolated From Seals.

Marine mammals are frequently considered good sentinels for human, animal and environmental health due to their long lifespan, coastal habitat, and characteristics as top chain predators. Using a One Health approach, marine mammals can provide information that helps to enhance the understanding of the health of the marine and coastal environment. Antimicrobial resistance (AMR) is the quintessential One Health problem that poses a well-recognised threat to human, animal, and ecosystem health worldwide. Treated and untreated sewage, hospital waste and agricultural run-off are often responsible for the spread of AMR in marine and freshwater ecosystems. Rescued seals (n = 25) were used as sentinels to investigate the levels of AMR in the Irish coastal ecosystem. Faecal swabs were collected from these animals and bacterial isolates (E. coli and cefotaxime-resistant non-E. coli) from each swab were selected for further investigation. E. coli isolates were characterised in terms of phylogenetic group typing, AMR, and virulence factors. All E. coli isolates investigated in this study (n = 39) were ampicillin resistant while 26 (66.6%) were multi-drug resistant (MDR). Resistance genes bla OXA-1 and bla TEM-1 were detected in 16/39 and 6/39 isolates, respectively. Additionally, virulence factors associated with adhesion (sfa, papA, and papC) and siderophores (fyuA and iutA) were identified. An additional 19 faecal cefotaxime-resistant non-E. coli isolates were investigated for the presence of ß-lactamase encoding genes. These isolates were identified as presumptive Leclercia, Pantoea and Enterobacter, however, none were positive for the presence of the genes investigated. To the authors knowledge this is the first study reporting the detection of bla OXA-1 and bla TEM-1 in phocid faecal E. coli in Europe. These results highlight the importance of marine mammals as sentinels for the presence and spread of AMR in the marine and coastal environment.

Financial Analysis of Herd Status and Vaccination Practices for Porcine Reproductive and Respiratory Syndrome Virus, Swine Influenza Virus, and Mycoplasma hyopneumoniae in Farrow-to-Finish Pig Farms Using a Bio-Economic Simulation Model.

This study aimed (1) to quantify the effects of positive status and vaccination practices for porcine reproductive and respiratory syndrome virus (PRRSv), swine influenza virus (SIV) and Mycoplasma hyopneumoniae (MHYO) on the profitability of farrow-to-finish pig farms and (2) to examine the financial impact of vaccination status in PRRSv and SIV positive farms. Data from 56 Irish farrow-to-finish pig farms were used for this study. Production effects associated with herd status for the three pathogens were incorporated into the Teagasc Pig Production Model (TPPM), a bio-economic stochastic simulation model for farrow-to-finish pig farms. In the analysis, farms negative (-) for either PRRSv, SIV or MHYO were assumed as baseline when presenting results for farms positive (+) for each pathogen. While all MHYO(+) farms used vaccination against the pathogen, not all PRRSv(+) or SIV(+) farms vaccinated against the disease. For all scenarios, a 728-sow farrow-to-finish farm with weekly farrowing batches was simulated. Financial risk analysis was conducted by Monte Carlo simulation within the TPPM using the Microsoft Excel add-in @Risk. Mortality rates, feedstuff costs and price per kg of meat produced were included as input stochastic variables and annual net profit was set as stochastic output variable. Positive farms sold fewer pigs and produced less kg of meat than negative farms and had increased feed usage during the weaner and finisher stages. Variable costs increased in positive farms due to increased feed costs, more dead animals for disposal and healthcare costs. Annual mean profit was lower by 24% in vaccinated PRRSv(+), 14.6% in unvaccinated PRRSv(+), 36.7% in vaccinating SIV(+), 12.8% in unvaccinated SIV(+), and 41% in MHYO(+) farms. Negative farms were first order stochastically dominant over positive farms, indicating that for a given level of profit, the financial risk is lower by avoiding respiratory pathogens. Similarly, unvaccinated farms were second order stochastically dominant over vaccinating farms suggesting that farms that do not vaccinate are less affected by the disease. Results from this study provide further evidence to encourage farmers to undertake improved disease control measures and/or to implement eradication programs.

Quantification, description and international comparison of antimicrobial use on Irish pig farms.

BACKGROUND: There is concern that the use of antimicrobials in livestock production has a role in the emergence and dissemination of antimicrobial resistance in animals and humans. Consequently, there are increasing efforts to reduce antimicrobial use (AMU) in agriculture. As the largest consumer of veterinary antimicrobials in several countries, the pig sector is a particular focus of these efforts. Data on AMU in pig production in Ireland are lacking. This study aimed to quantify AMU on Irish pig farms, to identify the major patterns of use employed and to compare the results obtained to those from other published reports and studies. RESULTS: Antimicrobial use data for 2016 was collected from 67 Irish pig farms which represented c. 35% of national production. The combined sample population consumed 14.5 t of antimicrobial by weight of active ingredient suggesting that the pig sector accounted for approximately 40% of veterinary AMU in Ireland in 2016. At farm level, median AMU measured in milligram per population correction unit (mg/PCU) was 93.9 (range: 1.0-1196.0). When measured in terms of treatment incidence (TI200), median AMU was 15.4 (range: 0.2-169.2). Oral treatments accounted for 97.5% of all AMU by weight of active ingredient and were primarily administered via medicated feed to pigs in the post weaning stages of production. AMU in Irish pig production in 2016 was higher than results obtained from the national reports of Sweden, Denmark, the Netherlands and France but lower than the United Kingdom. CONCLUSIONS: Pig production in Ireland is an important consumer of veterinary antimicrobials. The quantities and patterns of AMU on Irish pig farms are comparable to pig production in other European countries but higher than some countries with more advanced AMU reduction strategies. This AMU is characterised by a high proportion of prophylactic use and is primarily administered to pigs post weaning via medicated feed. Further studies to better understand the reasons for AMU on Irish pig farms and strategies to improve health among weaner pigs will be of benefit in the effort to reduce AMU.

Clinical presentation and immune characteristics in first-lactation Holstein-Friesian cows following intramammary infection with genotypically distinct Staphylococcus aureus strains.

Staphylococcus aureus is an important cause of bovine mastitis, and intramammary infections caused by this pathogen are often characterized as mild, chronic, or persistent. The strains of Staph. aureus associated with mastitis belong to several distinct bovine-adapted bacterial lineages. Studies of host-pathogen interactions have demonstrated that significant differences exist between Staph. aureus strains and lineages in their ability to internalize and to elicit expression of chemokines and pro-inflammatory mediators in bovine cells in vitro. To determine the effect of bacterial strain on the response to intramammary infection in vivo, 14 disease-free, first-lactation cows were randomly allocated to 2 groups and challenged with Staph. aureus strain MOK023 (belonging to CC97) or MOK124 (belonging to CC151). Clinical signs of infection, as well as somatic cell count (SCC), bacterial load, IL-8 and IL-1ß in milk, anti-Staph. aureus IgG in milk and serum, anti-Staph. aureus IgA in milk, and white blood cell populations in milk and blood were monitored for 30 d after the challenge. Cows infected with MOK023 generally developed subclinical mastitis, whereas cows infected with MOK124 generally developed clinical mastitis. Milk yield was reduced to a greater extent in response to infection with MOK124 compared with MOK023 in the first week of the study. Significantly higher SCC, IL-8, and IL-1ß in milk as well as higher anti-Staph. aureus IgG and IgA in milk and anti-Staph. aureus IgG in serum were also observed in response to MOK124 compared with the response to MOK023. Higher proportions of neutrophils were observed in milk of animals infected with MOK124 than in animals infected with MOK023. Higher neutrophil concentration in blood was also observed in the MOK124 group compared with the MOK023 group. Overall, the results indicate that the outcome of mastitis mediated by Staph. aureus is strain dependent.

Porcine reproductive and respiratory syndrome virus: phylogenetic analysis of circulating strains in the Republic of Ireland from 2016 to 2017.

In order to investigate the genetic diversity of porcine reproductive and respiratory syndrome virus (PRRSV) strains currently circulating in the Republic of Ireland (ROI), the ORF5 gene from 17 field strains originating from four vaccinating commercial herds was sequenced and phylogenetically analysed. High genetic variability was observed between farms at the nucleotide (86.3-95.2%) and amino acid (85.5-96%) levels. Phylogenetic analysis confirmed that all field strains belonged to the European species (type 1) and clustered into three separate groups within the subtype 1 subgroup. This variation may pose challenges for diagnosis and prophylactic control of PRRSV through vaccination in the ROI.

Influence of the Intestinal Microbiota on Colonization Resistance to Salmonella and the Shedding Pattern of Naturally Exposed Pigs.

Salmonella colonization and infection in production animals such as pigs are a cause for concern from a public health perspective. Variations in susceptibility to natural infection may be influenced by the intestinal microbiota. Using 16S rRNA compositional sequencing, we characterized the fecal microbiome of 15 weaned pigs naturally infected with Salmonella at 18, 33, and 45 days postweaning. Dissimilarities in microbiota composition were analyzed in relation to Salmonella infection status (infected, not infected), serological status, and shedding pattern (nonshedders, single-point shedders, intermittent-persistent shedders). Global microbiota composition was associated with the infection outcome based on serological analysis. Greater richness within the microbiota postweaning was linked to pigs being seronegative at the end of the study at 11 weeks of age. Members of the Clostridia, such as Blautia, Roseburia, and Anaerovibrio, were more abundant and part of the core microbiome in nonshedder pigs. Cellulolytic microbiota (Ruminococcus and Prevotella) were also more abundant in noninfected pigs during the weaning and growing stages. Microbial profiling also revealed that infected pigs had a higher abundance of Lactobacillus and Oscillospira, the latter also being part of the core microbiome of intermittent-persistent shedders. These findings suggest that a lack of microbiome maturation and greater proportions of microorganisms associated with suckling increase susceptibility to infection. In addition, the persistence of Salmonella shedding may be associated with an enrichment of pathobionts such as Anaerobiospirillum. Overall, these results suggest that there may be merit in manipulating certain taxa within the porcine intestinal microbial community to increase disease resistance against Salmonella in pigs. IMPORTANCE Salmonella is a global threat for public health, and pork is one of the main sources of human salmonellosis. However, the complex epidemiology of the infection limits current control strategies aimed at reducing the prevalence of this infection in pigs. The present study analyzes for the first time the impact of the gut microbiota in Salmonella infection in pigs and its shedding pattern in naturally infected growing pigs. Microbiome (16S rRNA amplicon) analysis reveals that maturation of the gut microbiome could be a key consideration with respect to limiting the infection and shedding of Salmonella in pigs. Indeed, seronegative animals had higher richness of the gut microbiota early after weaning, and uninfected pigs had higher abundance of strict anaerobes from the class Clostridia, results which demonstrate that a fast transition from the suckling microbiota to a postweaning microbiota could be crucial with respect to protecting the animals.

The in vitro host cell immune response to bovine-adapted Staphylococcus aureus varies according to bacterial lineage.

Mastitis is the most economically important disease affecting dairy cattle worldwide. Staphylococcus aureus is a highly prevalent cause of mastitis, causing infections ranging from sub-clinical to gangrenous. However, the interaction between the genotype of the infecting strain of S. aureus and the host response remains largely uncharacterised. To better understand the variation in presentation and outcomes of S. aureus-mediated bovine mastitis, we studied the interaction of a panel of mastitis isolates from several prominent bovine-associated lineages with bovine mammary epithelial cells (bMEC) and neutrophils. Significant differences in immune gene expression by infected primary or immortalised bMEC, or their elaboration of neutrophil chemoattractants, were observed and were dependent on the lineage of the infecting strain. Differences were also apparent in the invasiveness of S. aureus strains and their ability to survive killing by neutrophils. Our results demonstrate that a range of immune responses occur, suggesting the importance of S. aureus strain in dictating mastitis disease course. S. aureus lineages may therefore have adopted differing strategies for exploitation of the intramammary niche. Consequently, improved diagnosis of infecting lineage may enable better prognosis for S. aureus mastitis and reduce morbidity and economic loss.