Ian Robertson

This study aimed to determine the prevalence and associated plasmid resistant genome of extended spectrum β-lactamase (ESBL) and Ambler class C cephalosporinase (AmpC) producing commensal Escherichia coli, using a targeted pDNA approach, from lactating cows and pre-weaned calves on dairy farms in Western Australian (WA). Following culture and modified double disc diffusion, ESBL and/or AmpC producing commensal E. coli were isolated from faecal samples and phenotypic antimicrobial resistance (AMR) determined. Plasmid DNA (pDNA) was extracted, amplified, and sequenced to identify the AMR and replicon composition. Phenotypic analysis of 1,117 faecal samples from 26 Western Australian dairy farms revealed a low overall prevalence (7.3%) of ESBL and/or AmpC-producing commensal E. coli, with 3.2% in cows (n = 633) and 12.8% in calves (n = 484). All isolates were classified ‘non-wildtype’ for ampicillin and ceftiofur, with 95.1% classified as ‘non-wildtype’ multidrug resistant (MDR). While blaTEM was common in the pDNA (total: 60.6%; cow: 76.9%; calf: 57.7%), a total of 13 AMR genes were identified across 126 plasmids (cow: n = 30; calf: n = 96) isolated. Multiple correspondence analysis (with chi-square–based 95% confidence ellipses) showed variation in pDNA AMR genes and replicons by farm, with generally distinct plasmid profiles, though some clustering suggests possible farm-to-farm plasmid transmission. In conclusion, though low in prevalence, ESBL and/or AmpC producing commensal E. coli having high phenotypic AMR, MDR, and pDNA diversity were isolated from dairy cattle on 65.4% of screened farms.
•Low prevalence of commensal E. coli producing ESBL and/or AmpC enzymes.•ESBL and/or AmpC producing commensal E. coli were isolated from dairy cattle on 65.4% of the 26 farms screened.•Isolates had high phenotypic resistance, multiple drug resistance, and plasmid diversity.•Targeted pDNA approach appreciates AMR diversity with lower cost, time, and data complexity.

A (SVA) is an emerging threat to swine populations due to its potential to cause vesicular lesions, which are difficult to differentiate from other vesicular diseases of swine such as foot and mouth disease (FMD), requiring significant resources for differential diagnosis. The first Taiwanese isolate of SVA was identified in 2006, although the first clinical case was not reported until 2012. The genetic characteristics and seroprevalence of SVA in Taiwan remain unclear. This study aimed to assess the seroprevalence and genetic diversity of SVA in nursery/weaned swine and finisher swine on Taiwanese pig farms. Phylogenetic analysis of seven Taiwanese SVA isolates revealed clustering into groups I and II. The 2006 and 2012 isolates shared 95.5% and 95.7% identity, respectively, with an early USA strain (MT360258), while more recent strains collected between 2018 and 2022 exhibited 95.7-98.8% identity with a 2020 USA strain (MZ733977). Serological analysis of swine from 300 farms showed significantly higher herd-level seroprevalence in nursery/weaned swine (53%) than finisher swine (6.7%). Furthermore, comparative analysis of nine known B cell epitopes showed high sequence conservation across Taiwanese and global strains. These findings provide important baseline data on the genetic diversity and seroprevalence of SVA in Taiwan and support the development of improved surveillance strategies for this emerging swine pathogen.

Brucellosis, a zoonotic disease primarily transmitted through livestock, poses significant health and economic challenges globally. Its increasing incidence in China highlights critical gaps in current control measures, particularly in regions with high livestock movement, where domestic ruminants trade networks elevate the risk of disease transmission. Traditional value chain analyses often provide descriptive mappings of these networks but lack quantitative methods to assess specific risk levels. This study integrates value chain with quantitative risk assessment using Monte Carlo simulations to evaluate the risk of brucellosis introduction via cattle imports in Zhijiang. We comprehensively mapped the local cattle value chain, identifying key stakeholders, risk nodes, and critical pathways of infection. Utilizing a stochastic scenario tree model based on local data, we estimated the likelihood of infected cattle entering the region, revealing that unregulated brokers and inadequate quarantine measures significantly contribute to the risk. Our sensitivity analysis confirmed vulnerabilities in existing biosecurity protocols, emphasizing the need for enhanced quarantine measures and reliable testing at high-risk points in cattle trade. By combining value chain insights with quantitative analysis, this study effectively identifies critical vulnerabilities within livestock networks. Our findings lay the groundwork for developing targeted brucellosis prevention strategies in Zhijiang, underscoring the urgent need for strengthened biosecurity and risk management practices.

Caprine brucellosis, mainly caused by Brucella melitensis, remains a significant zoonotic threat worldwide, affecting animal productivity, welfare, and public health. This study aimed to estimate the true prevalence (TP) and spatial distribution of antibodies to Brucella spp. among goat populations in Hubei Province, China. In 2021, approximately 1.4 million serum samples were collected from 23,126 goat flocks across 82 counties of 16 municipal regions of Hubei Province. A combination of the Rose Bengal Test and Serum Agglutination Test in series was used to detect antibodies against Brucella spp. A hierarchical Bayesian Latent Class Model was used to account for imperfect diagnostic sensitivity and specificity of the tests, conditional dependence between the two tests, and hierarchical data structure to estimate the TP and the probability of achieving a 95 % probability of having a TP below 0.1 % for each county and municipal region. Apparent prevalence was 0.051 % and 0.536 % at the animal and flock level, respectively. The median animal level TP in the 82 counties was 0.0088 % (Range: 0.0008 %, 9.3730 %), with 76.8 % of counties showing a median TP estimate below 0.1 %. Counties containing positive goats were mainly clustered in Huanggang and Huangshi, and counties bordering positive counties had a higher risk of seropositivity. Notably, 52.4 % of counties achieved a 95 % probability with a TP below 0.1 %. Sensitivity analyses confirmed the robustness of these findings across prior distributions. It was concluded that Hubei Province has achieved remarkable progress in caprine brucellosis elimination programs, and priority interventions should be given to positive counties and their bordering counties.

This study aimed to determine the prevalence of antimicrobial resistance (AMR) in commensal E. coli from healthy lactating cows and calves in the Mediterranean pasture-based feeding dairy system of Western Australia (WA). Fecal samples were collected from healthy adult lactating cows and healthy calves from dairy farms in WA. Presumptive commensal E. coli was isolated from these samples and confirmed using matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Broth microdilution was used to assess the prevalence and the phenotypic AMR profiles of the E. coli isolates to 8 antimicrobial agents of dairy industry and human importance. The minimum inhibitory concentration (MIC) for each isolate was interpreted using the Epidemiologic Cutoff (ECOFF) and Clinical and Laboratory Standards Institute (CLSI) breakpoints. Genomic characterization provided multi-locus sequence types and AMR genes for a selection of isolates categorised as non-wild type (NWT) by ECOFF values for the combination of ampicillin, trimethoprim-sulfamethoxazole, and tetracycline. From a total of 1,117 fecal samples (633 adult, 484 calf) collected across 26 randomly selected farms, 891 commensal E. coli isolates were recovered (541 adult, 350 calf). Commensal E. coli classified as NWT was highest for ampicillin for both adult (68.8%; 95% CI = 64.7 – 72.7) and calf feces (67.1%; 95% CI = 62.0 – 72.0). A large proportion of tetracycline NWT and trimethoprim-sulfamethoxazole NWT organisms were also identified from calf feces, being 44.0% (95% CI = 38.7 – 49.4) and 24.6% (95% CI = 20.2 – 29.4) respectively. Clinical resistance prevalence was low, being higher for calves than for adult feces (ampicillin (adult: 7.8% (95% CI = 5.7 – 10.3); calf: 30.0% (95% CI = 25.2 – 35.1), tetracycline (adult: 6.3% (95% CI = 4.4 – 8.7); calf: 40.3% (95% CI = 35.1 – 45.6), and trimethoprim-sulfamethoxazole (adult: 2.6% (95% CI = 1.4 – 4.3); calf: 22.0% (95% CI = 17.7 – 26.7)). Commensal E. coli originating from calf feces was significantly higher in NWT prevalence compared with adult feces for ciprofloxacin (P = 0.023), gentamicin (P = 0.02), tetracycline (P < 0.001), and trimethoprim-sulfamethoxazole (P < 0.001). The overall number of antimicrobials an isolate was classified as NWT toward varied among farms and was significantly higher for isolates originating from calf than adult feces (P < 0.001). The strain type and sampling source of the commensal E. coli investigated were both associated with the commonality of the resultant resistance genome. Clinical resistance and NWT classification were highest for ampicillin, tetracycline, and trimethoprim-sulfamethoxazole, all antimicrobials commonly used in the treatment of dairy cattle in Australia. Though highly variable across farms, commensal E. coli isolated from healthy dairy calf feces had significantly higher NWT and multidrug resistance (MDR) prevalence compared with feces from healthy adult lactating dairy cows. The resistant genome identified in MDR isolates, though not always consistent with the phenotype, included QnrS1 and genes encoding AmpC β-lactamase and aminoglycoside phosphotransferase.

Enzootic bovine leukosis, a neoplastic disease caused by the bovine leukemia virus (BLV), was the primary cancer affecting cattle in China before 1985. Although its prevalence decreased significantly between 1986 and 2000, enzootic bovine leukosis has been re-emerging since 2000. This re-emergence has been largely overlooked, possibly due to the latent nature of BLV infection or the perceived lack of sufficient evidence. This study investigated the molecular epidemiology of BLV infections in dairy cattle in Henan province, Central China. Blood samples from 668 dairy cattle across nine farms were tested using nested polymerase chain reaction assays targeting the partial envelope (
) gene (gp51 fragment). Twenty-three samples tested positive (animal-level prevalence of 3.4%; 95% confidence interval: 2.2, 5.1). The full-length
gene sequences from these positive samples were obtained and phylogenetically analyzed, along with previously reported sequences from the GenBank database. The sequences from positive samples were clustered into four genotypes (1, 4, 6, and 7). The geographical annotation of the maximum clade credibility trees suggested that the two genotype 1 strains in Henan might have originated from Japan, while the genotype 7 strain is likely to have originated from Moldova. Subsequent Bayesian stochastic search variable selection analysis further indicated a strong geographical association between the Henan strains and Japan, as well as Moldova. The estimated substitution rate for the
gene ranged from 4.39 × 10
to 2.38 × 10
substitutions per site per year. Additionally, codons 291, 326, 385, and 480 were identified as positively selected sites, potentially associated with membrane fusion, epitope peptide vaccine design, and transmembrane signal transduction. These findings contribute to the broader understanding of BLV epidemiology in Chinese dairy cattle and highlight the need for measures to mitigate further BLV transmission within and between cattle herds in China.

Mycobacterium avium subsp. paratuberculosis (MAP) is responsible for the persistent infectious illness known as bovine paratuberculosis, which is one of the most easily overlooked diseases in China amid a lack of epidemiological data. In this study, we evaluated the agreement of milk and blood antibody tests for paratuberculosis and showed an overall agreement of 92.0%, with a 95.0% negative coincidence rate and a 78.6% positive coincidence rate. The milk test was then used to examine the prevalence and incidence of dairy cows in Hubei Province, China. We found that, at the individual level, the highest lacto-prevalence reached up to 22.9%; the farm-level prevalence was as high as 92.3% (12/13) and 84.6% (11/13) in January and April 2018, respectively. The total incidence risk of all farms was 6% per three months. We also found that large-scale farms had a significantly lower prevalence and incidence than small-scale farms. Finally, the correlation between paratuberculosis and milk quality was evaluated, and we confirmed that MAP can significantly alter milk quality and raise somatic cell counts in the milk. This study provides valuable information for assessing the prevalence and incidence risk of paratuberculosis in China. It further provides an essential basis for calling for the prevention and control of paratuberculosis in China.

Quantification of antimicrobial resistance (AMR) is beneficial to inform policies and direct prudent antimicrobial use.

This study aimed to assess the current published evidence of AMR from passive and active ad hoc surveillance activities within the Australian dairy cattle industry.
Following a scoping review framework 373 articles published before January 2023 were retrieved using the keyword search function from two online databases (PubMed® and Web of Science™ Core Collection). The duplicate articles were removed and the title, abstract, and full text of the remaining articles were reviewed following the study objectives and inclusion criteria (location, subject/theme, and data). Data from the remaining articles were extracted, summarised, interpreted and the study quality assessed using the Grades of Recommendations, Assessment, Development, and Evaluation guidelines.

A total of 29 articles dating from the 1960 s until 2022 were identified to meet the study criteria (passive: n = 15; active: n = 14). Study characteristics such as sampling type, sampling method, and AMR assessment were all common characteristics from both passive and active surveillance articles, being milk samples, individual sampling, and phenotypic assessment respectively. Passive surveillance articles had a wider range in both the type of bacteria and the number of antimicrobials investigated, while active surveillance articles included a higher number of bacterial isolates and sampling from healthy populations. There was an overall low level of clinical AMR across all articles. Higher prevalence of non-wildtype Escherichia coli, Salmonella spp., and Staphylococcus spp., although limited in data, was suggested for commonly used Australian veterinary antimicrobials for these bacteria. The prevalence of phenotypic AMR varied due to the health and age status of the sampled animals. The articles reviewed in this study suggest the prevalence of AMR genes was higher for commonly used antimicrobials, although genes were not always related to the phenotypic AMR profile.

Published evidence of AMR in the Australian dairy cattle industry is limited as demonstrated by only 29 articles included in this review following selection criteria screening. However, collectively these articles provide insight on industry AMR prevalence. For example, the suggestion of non-wildtype bacteria within the Australian dairy cattle indicating a risk of emerging or increasing industry AMR. Therefore, further surveillance is required to monitor the development of future AMR risk within the industry. Additionally, evidence suggesting that animals varying in health and age differ in prevalence of AMR imply a requirement for further research into animal population demographics to reduce potential bias in data collated in both national and global surveillance activities.

Dairy cattle and their products have been linked to human outbreaks of Salmonella and Escherichia coli (E. coli). The objective of this study was to estimate the prevalence and antimicrobial resistance of Salmonella and to enumerate Extended-spectrum β-lactamase (ESBL)-producing E. coli in apparently healthy lactating dairy cows in Hubei Province, China. In a cross-sectional study, a total of 291 adult lactating dairy cows from 10 dairy farms were sampled for the detection of Salmonella and ESBL E. coli. Overall, Salmonella was cultured from 15 out of 291 sampled animals (5.2 %; 95 % confidence intervals (CI): 2.9, 8.4), all from two herds with a herd prevalence of 20.0 % (95 % CI: 2.5, 55.6) and the main serotype being S. Dublin. The within-herd prevalence ranged between 0.0 % and 33.3 %. ESBL E. coli was detected by culture in all farms with an animal level prevalence of 59.1 % (95 % CI: 53.2, 64.8) and 116 samples (39.9 %, 95 % CI: 34.2, 45.7) contained ESBL E. coli with a number exceeding 10
CFU/g feces. Sixty percent (9/15) of Salmonella isolates were resistant to ampicillin, however all isolates were sensitive to the other 8 antimicrobials tested. Ninety percent (95 % CI: 84.6, 94.1) of ESBL E. coli contained the resistance gene bla
, but no ESBL Salmonella was found. Our findings contribute to the understanding of the prevalence and antimicrobial resistance of Salmonella and the enumeration of ESBL E. coli and will assist in the decision-making for the control of Salmonella in Hubei Province.

Background
Tuberculosis (TB) is a zoonotic disease that affects humans and domesticated and wild animals. Animals in zoos are potentially an important source of TB for humans; however they are often neglected in routine disease surveillance programs. This investigation reports an outbreak of TB in milu deer and northern pig-tailed macaques in a zoo in Wuhan, China, which highlighted the need for improved prevention and control of TB in China.

Methods
Between 24 November and 9 December 2020 two milu deer and a northern pig-tailed macaque that were displaying signs of wasting died. Post-mortem, histopathological diagnosis and acid fast staining were used for the dead animals. Multiple PCR for Mycobacterium tuberculosis complex (MTBC) was performed to identify the bacterial in both milu deer and northern pig-tailed macaque. The serum antibody iELISA for MTBC was then performed for all the surviving milu deer and northern pig-tailed macaques. Six seropositive milu deer and a seropositive northern pig-tailed macaque were subsequently euthanised and, along with two other dead milu deer, necropsied. DNA from these tissue samples was extracted and detected MTBC using PCR and Real-time PCR. Subsequently bacterial isolation was used to confirm the infection.

Results
The lungs of the dead animals displayed gross and histological TB-like lesions and changes, and red staining bacilli were detected in smears of the lesions by microscopy after acid fast staining. Mycobacterium bovis (M. bovis) was detected in the two milu deer and Mycobacterium tuberculosis (M. tb) in the northern pig-tailed macaque using multiple PCR for MTBC. 35.3% surviving milu deer and 50% surviving northern pig-tailed macaques MTBC serologically positive. Six of the euthanised milu deer were also positive on a DNA test for M. bovis and the euthanised northern pig-tailed macaque was positive to M. tb.

Conclusions
This is the first report of tuberculosis in the endangered species, milu deer and northern pig-tailed macaques, in China, and warrants urgent attention by researchers and conservation authorities. These cases highlight the need for expanding surveillance for MTBC to zoos in China.