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Antibiotics (AB) are used in intensive pig production systems to control infectious diseases and they are suspected to be a major source of antibiotic resistance. Following the ban on AB use as growth promoters in the EU, their prophylactic use in-feed is now under review. The aim of this study was to evaluate the effect of removing prophylactic in-feed AB on pig health and welfare indicators. Every Monday for six weeks, a subset of 70 pigs were weaned, tagged and sorted into two groups of 35 pigs according to weight (9.2 ± 0.6 kg). AB were removed from the diet of one group (NO, n = 6) and maintained in the other group (AB, n = 6) for nine weeks. Ten focal pigs were chosen per group. After c. five weeks each group was split into two pens of c.17 pigs for the following 4 weeks. Data were recorded weekly. Skin, tail, ear, flank and limb lesions of focal pigs were scored according to severity. The number of animals per group affected by health deviations was also recorded. The number of fights and harmful behaviours (ear, tail bites) per group was counted during 3×5min observations once per week. Data were analysed using mixed model equations and binomial logistic regression. At group level, AB pigs were more likely to have tail (OR = 1.70; P = 0.05) but less likely to have ear lesions than NO pigs (OR = 0.46; P<0.05). The number of ear bites (21.4±2.15 vs. 17.3±1.61; P<0.05) and fights (6.91±0.91 vs. 5.58±0.72; P = 0.09) was higher in AB than in NO pigs. There was no effect of treatment on health deviations and the frequency of these was low. Removing AB from the feed of weaner pigs had minimal effects on health and welfare indicators.

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.

Background Gliomas in dogs remain poorly understood. Objectives To characterize the clinicopathologic findings, diagnostic imaging features and survival of a large sample of dogs with glioma using the Comparative Brain Tumor Consortium diagnostic classification. Animals Ninety‐one dogs with histopathological diagnosis of glioma. Methods Multicentric retrospective case series. Signalment, clinicopathologic findings, diagnostic imaging characteristics, treatment, and outcome were used. Tumors were reclassified according to the new canine glioma diagnostic scheme. Results No associations were found between clinicopathologic findings or survival and tumor type or grade. However, definitive treatments provided significantly (P = .03) improved median survival time (84 days; 95% confidence interval [CI], 45‐190) compared to palliative treatment (26 days; 95% CI, 11‐54). On magnetic resonance imaging (MRI), oligodendrogliomas were associated with smooth margins and T1‐weighted hypointensity compared to astrocytomas (odds ratio [OR], 42.5; 95% CI, 2.42‐744.97; P = .04; OR, 45.5; 95% CI, 5.78‐333.33; P < .001, respectively) and undefined gliomas (OR, 84; 95% CI, 3.43‐999.99; P = .02; OR, 32.3; 95% CI, 2.51‐500.00; P = .008, respectively) and were more commonly in contact with the ventricles than astrocytomas (OR, 7.47; 95% CI, 1.03‐53.95; P = .049). Tumor spread to neighboring brain structures was associated with high‐grade glioma (OR, 6.02; 95% CI, 1.06‐34.48; P = .04). Conclusions and Clinical Importance Dogs with gliomas have poor outcomes, but risk factors identified in survival analysis inform prognosis and the newly identified MRI characteristics could refine diagnosis of tumor type and grade.

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.

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 bla CTX-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.

Background Procalcitonin (PCT) is a widely used biomarker of sepsis in human medicine and can have potential applications in the veterinary field. This study aimed to explore whether PCT could be measured in the saliva of pigs and whether its concentration changes in sepsis. Therefore, a specific assay was developed and analytically validated, and changes in PCT concentration were evaluated in two conditions: a) in an experimental model of sepsis produced by the administration of lipopolysaccharide (LPS) to pigs ( n  = 5), that was compared with a model of non-septic inflammation induced by turpentine oil ( n  = 4), and b) in healthy piglets ( n  = 11) compared to piglets with meningitis ( n  = 20), a disease that usually involves sepsis and whose treatment often requires large amounts of antibiotics in farms. Results The assay showed coefficients of variation within the recommended limits and adequate linearity after serial sample dilutions. The method's detection limit was set at 68 μg/L, and the lower limit of quantification was 414 μg/L. In the LPS experiment, higher concentrations of PCT were found after 24 h in the animals injected with LPS (mean = 5790 μg/L) compared to those treated with turpentine oil (mean = 2127 μg/L, P  = 0.045). Also, animals with meningitis had higher concentrations of PCT (mean = 21515 μg/L) than healthy pigs (mean = 6096 μg/L, P value < 0.0001). Conclusions According to these results, this assay could be potentially used as a tool for the non-invasive detection of sepsis in pigs, which is currently a topic of high importance due to antibiotic use restriction.

Background Antibiotics (AB) are an important tool to tackle infectious disease in pig farms; however some research indicates that their frequent mis/over-use may contribute to the development of antibiotic resistance and the WHO has declared that this issue should be addressed. Little is known about the long term consequences of withdrawing prophylactic AB from pig feed; hence we aimed to assess its effects on performance and health of pigs from weaning to slaughter. Six batches of 140 pigs each were monitored on a commercial farm through the weaner and finisher stages to slaughter. In-feed antibiotics were not added to the feed for half of the pigs (NOI) and were added in the other half (ABI) within each batch for the whole weaner stage. Individual pigs in both treatments were treated with parenteral administrations if and when detected as ill or lame. Productive performance, parenteral treatments and mortality were recorded on farm and the presence of respiratory disease was recorded at slaughter. Pen was considered the experimental unit. Results ABI pigs showed higher growth ( P  = 0.018) and feed intake ( P  = 0.048) than NOI pigs in the first weaner stage but feed efficiency was not affected (NOI = 1.48 vs. ABI = 1.52). Despite an initial reduction in performance, NOI pigs had similar performance in finisher stage (ADG: NOI = 865.4 vs. ABI = 882.2) and minimal effects on health compared to ABI pigs. No difference between treatments was found at the abattoir for the percentage of pigs affected by pneumonia, pleurisy, pleuropneumonia and abscesses ( P  > 0.05). Mortality rate was not affected by treatment during the weaner stage ( P  = 0.806) although it tended to be slightly higher in NOI than ABI pigs during the finisher stage ( P  = 0.099). Parenteral treatments were more frequent in NOI pigs during the weaner stage ( P  <  0.001) while no difference was recorded during the finisher stage ( P  = 0.406). Conclusions These data suggest that the removal of prophylactic in-feed antibiotics is possible with only minor reductions in productive performance and health which can be addressed by improved husbandry and use of parenteral antibiotics.

Post-weaning diarrhoea (PWD) is a multifactorial disease that affects piglets after weaning, contributing to productive and economic losses. Its control includes the use of in-feed prophylactic antibiotics and therapeutic zinc oxide (ZnO), treatments that, since 2022, are no longer permitted in the European Union due to spread of antimicrobial resistance genes and pollution of soil with heavy metals. A dysbiosis in the microbiota has been suggested as a potential risk factor of PWD onset. Understanding pig's microbiota development around weaning and its changes in response to ZnO and antibiotics is crucial to develop feasible alternatives to prophylactic and metaphylactic antimicrobial use. This study used shotgun metagenomic sequencing to investigate the environmental and faecal microbiota on 10 farms using (Treated) or not using (ZnO-free) in-feed antibiotics and ZnO during the first 14 days post-weaning (dpw). Environmental samples from clean pens were collected at weaning day (0dpw), and faecal samples at 0, 7 and 14dpw. Diarrhoeic faecal samples were collected at 7dpw when available. The analysis of data revealed that the faecal microbiota composition and its functionality was impacted by the sampling time point (microbiota maturation after weaning) but not by the farm environment. Treatment with antibiotics and ZnO showed no effects on diversity indices while the analyses of microbiota taxonomic and functional profiles revealed increased abundance of taxa and metabolic functions associated with or different species of . on the Treated farms, and with and on the ZnO-free farms. The analysis of diarrhoea samples revealed that the treatment favoured the microbiota transition or maturation from 0dpw to 14dpw in Treated farms, resembling the composition of healthy animals, when compared to diarrhoea from ZnO-free farms, which were linked in composition to 0dpw samples. The results provide a comprehensive overview of the beneficial effects of ZnO and antibiotics in PWD in the microbiota transition after weaning, preventing the overgrowth of pathogens such as pathogenic and revealing the key aspects in microbiota maturation that antibiotics or ZnO alternatives should fulfil.

Background Antimicrobial resistance (AMR) is a global public health threat consequence of antimicrobial use (AMU) in human and animal medicine. In food-producing animals factors such as management, husbandry or biosecurity may impact AMU. Organic and extensive Iberian swine productions are based on a more sustainable and eco-friendly management system, providing an excellent opportunity to evaluate how sustained differences in AMU impact the AMR in indicator bacteria. Here, we evaluate the usefulness of commensal Escherichia coli and Enterococcus spp. isolates as AMR bioindicators when comparing 37 Spanish pig farms from both intensive and organic-extensive production systems, considering the effect of AMU and biosecurity measures, the last only on intensive farms. Results The production system was the main factor contributing to explain the AMR differences in E. coli and Enterococcus spp. In both bacteria, the pansusceptible phenotype was more common ( p  < 0.001) on organic-extensive farms when compared to intensive herds. The microbiological resistance in commensal E. coli was, for most of the antimicrobials evaluated, significantly higher ( p  < 0.05) on intensive farms. In enterococci, the lincosamides usage revealed the association between AMR and AMU, with an increase in the AMR for erythromycin ( p  < 0.01), quinupristin-dalfopristin ( p <  0.01) and the multidrug-resistant (MDR) phenotype ( p  < 0.05). The biosecurity measures implemented on intensive farms influenced the AMR of these bioindicators, with a slightly lower resistance to sulfamethoxazole ( p  < 0.01) and the MDR phenotype ( p  < 0.05) in E. coli isolated from farms with better cleaning and disinfection protocols. On these intensive farms, we also observed that larger herds had a higher biosecurity when compared to smaller farms ( p  < 0.01), with no significant associations between AMU and the biosecurity scores. Conclusions Overall, this study evidences that the production system and, to a lesser extent, the biosecurity measures, contribute to the AMR development in commensal E. coli and Enterococcus spp., with antimicrobial usage as the main differential factor, and demonstrates the potential value of these bacteria as bioindicators on pig farms in AMR surveillance programs.

[...]liberally comparing outcomes and survival after radiotherapy in dogs with unconfirmed glioma with surgery in histologically confirmed cases 5 is not accurate and could mislead readers. By contrast, a subsequently published study including 33 gliomas, all treated with surgical resection and immunotherapy, identified astrocytomas and low-grade tumors were associated with increased survival. 8 Whereas the latter study findings result from univariable analysis ours resulted from univariable followed by multivariable Cox proportional hazard modeling. [...]differences between both studies' findings might be attributable to methodology of statistical analysis and lack of homogeneity of treatment modalities. [...]evidence based on larger case numbers and multivariable statistical analysis is necessary to confirm any association between glioma type or grade and prognosis in dogs. [...]we take this opportunity once again to invite researchers to contribute to the creation of a mutually accessible international multicenter database to better enable evidence-based research in the field of canine glioma.