Journal Articles
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Item Zoonotic transmission of asymptomatic carriage Staphylococcus aureus on dairy farms in Canterbury, New Zealand.(Microbiology Society, 2024-12-04) Straub C; Taylor W; French NP; Murdoch DR; Priest P; Anderson T; Scott PZoonotic pathogen transmission is of growing concern globally, with agricultural intensification facilitating interactions between humans, livestock and wild animals. Staphylococcus aureus is a major human pathogen, but it also causes mastitis in dairy cattle, leading to an economic burden on the dairy industry. Here, we investigated transmission within and between cattle and humans, including potential zoonotic transmission of S. aureus isolated from cattle and humans from three dairy farms and an associated primary school in New Zealand. Nasal swabs (N=170) were taken from healthy humans. Inguinal and combined nasal/inguinal swabs were taken from healthy cattle (N=1163). Whole-genome sequencing was performed for 96 S. aureus isolates (44 human and 52 cattle). Multilocus sequence typing and assessments of antimicrobial resistance and virulence were carried out. Potential within- and across-species transmission events were determined based on single nucleotide polymorphisms (SNPs). Thirteen potential transmission clusters were detected, with 12 clusters restricted to within-species and one potential zoonotic transmission cluster (ST5). Potential transmission among cattle was mostly limited to single age groups, likely because different age groups are managed separately on farms. While the prevalence of antimicrobial resistance (AMR) was low among both bovine and human isolates, the discovery of an extended-spectrum beta-lactamase gene (bla TEM-116) in a bovine isolate was concerning. This study provides evidence around frequency and patterns of potential transmission of S. aureus on dairy farms and highlights the AMR and virulence profile of asymptomatic carriage S. aureus isolates.Item Retention of internal teat sealants over the dry period and their efficacy in reducing clinical and subclinical mastitis at calving(Elsevier Inc and the Federation of Animal Science Societies (Fass) Inc on behalf of the American Dairy Science Association, 2022-06) Bates AJ; King C; Dhar M; Fitzpatrick C; Laven RAInternal teat sealants (ITS) reduce the risk of new intramammary infections over the dry period by forming a physical barrier to pathogen ingress. As the first and last 2 wk of the dry period are high-risk periods for new infections, maintaining an effective barrier in this period is a key requirement. Few studies have systematically examined sealant retention and none have done so under New Zealand pastoral conditions, where cows frequently move to separate grazing for dry periods, typically 80 to 90 d long. This multi-herd study was a split-udder equivalence trial comparing 2 ITS formulations for retention and efficacy in preventing periparturient clinical and subclinical mastitis. Both ITS contained 65% (2.6 g) bismuth salts, which contribute to the barrier within the teat canal, emulsified in ≤1.4 g of mineral oil. However, one ITS additionally contained <10% amorphous silica. At dry-off, treatment was randomly allocated to diagonal teat-pairs within 409 cows on 4 farms. All cows met industry best practice criteria for ITS treatment alone. The study unit was quarter within cow and farm. Outcomes included clinical mastitis (CM) incidence for the last 7 d of the dry period and first 42 d of lactation, subclinical mastitis (SCM) incidence 96 h after calving, and quantity of residual after centrifuging 50 mL of colostrum collected from each quarter within 24 h of calving. Proportional outcomes were analyzed using Bayesian mixed models with a binomial distribution and logit link function, whereas the quantity of residual was analyzed using Bayesian finite mixture models and cluster bootstrapping. We set a region of probable equivalence (ROPE) of ±2.5% between proportions and ±0.2 g for residual weight. Records were available for 1,596 quarters (399 cows). We detected no meaningful difference in incidence of CM or SCM attributable to differences in sealant: the model predicted treatment differences of 0.00 with a 95% highest density interval (HDI) of ±1.00%. Across all cows and farms, the marginal difference in the percentage of quarters with CM was 0.11% (95% HDI: -2.11 to 2.49%), and for SCM 0.00 (95% HDI: -1.98 to 1.94%). Including the quantity of residual recovered at calving did not improve fit or predictive ability of the models predicting CM or SCM, and the coefficient spanned the null value. The distribution of the weight of material recovered at calving was multi-modal; for 25% of quarters, more residual was recovered than inserted. When the residual weight was less than or equal to the median residual weight (2.06 g; range: 0.19-6.03 g), there was a ≥90% probability that any treatment difference in residual was ≤0.2 g. When the residual weight was between the median and 75th percentile (4.40 g; 95% HDI: 4.00 to 4.75 g), there was no clear difference in residual between products. Above the 75th percentile, there was a 90% probability that the residual from quarters differed by product type (difference = 0.36 g, 90% HDI: 0.20 to 0.54 g). In conclusion, both products had equivalent efficacy for SCM and CM. As the quantity of residual increased, the difference in residual weight recovered increased but this may represent increases in debris rather than indicating a more effective barrier.Item Microtiter Screening Reveals Oxygen-Dependent Antimicrobial Activity of Natural Products Against Mastitis-Causing Bacteria(Frontiers Media S.A., 2019-08-28) Ferguson SA; Menorca A; Van Zuylen EM; Cheung C-Y; McConnell MA; Rennison D; Brimble MA; Bodle K; McDougall S; Cook GM; Heikal A; Debnath AIn this study we investigated the influence of oxygen availability on a phenotypic microtiter screen to identify new, natural product inhibitors of growth for the bovine mastitis-causing microorganisms; Streptococcus uberis, Staphylococcus aureus, and Escherichia coli. Mastitis is a common disease in dairy cattle worldwide and is a major cause of reduced milk yield and antibiotic usage in dairy herds. Prevention of bovine mastitis commonly relies on the application of teat disinfectants that contain either iodine or chlorhexidine. These compounds are used extensively in human clinical settings and increased tolerance to chlorhexidine has been reported in both Gram-positive and Gram-negative microorganisms. As such new, non-human use alternatives are required for the agricultural industry. Our screening was conducted under normoxic (20% oxygen) and hypoxic (<1% oxygen) conditions to mimic the conditions on teat skin and within the mammary gland respectively, against two natural compound libraries. No compounds inhibited E. coli under either oxygen condition. Against the Gram-positive microorganisms, 12 inhibitory compounds were identified under normoxic conditions, and 10 under hypoxic conditions. Data revealed a clear oxygen-dependency amongst compounds inhibiting growth, with only partial overlap between oxygen conditions. The oxygen-dependent inhibitory activity of a naturally occurring quinone, β-lapachone, against S. uberis was subsequently investigated and we demonstrated that this compound is only active under normoxic conditions with a minimum inhibitory concentration and minimum bactericidal concentration of 32 μM and kills via a reactive oxygen species-dependent mechanism as has been demonstrated in other microorganisms. These results demonstrate the importance of considering oxygen-availability in high-throughput inhibitor discovery.Item Antimicrobial susceptibilities in dairy herds that differ in dry cow therapy usage(Elsevier Inc and the Federation of Animal Science Societies (Fass) Inc on behalf of the American Dairy Science Association, 2021-08) McDougall S; Penry J; Dymock DIntramammary infusion of antimicrobials at the end of lactation (dry cow therapy; DCT) is a central part of mastitis control programs and is one of the major indications for antimicrobial use in dairy cows. However, with increasing focus on prudent use of antimicrobials and concerns about emergence of antimicrobial resistance, the practice of treating every cow at the end of lactation with DCT is in question. This cross-sectional, observational study determined the minimum inhibitory concentrations (MIC) of 10 antimicrobials for coagulase-negative staphylococci (CNS), Staphylococcus aureus, Streptococcus dysgalactiae, and Streptococcus uberis isolates from milk samples from dairy cows with somatic cell counts >200,000 cells/mL in herds that had been organic for >3 yr (n = 7), or had used either ampicillin-cloxacillin DCT (n = 11) or cephalonium DCT (n = 8) in the preceding 3 yr. The organic herds were certified under the United States Department of Agriculture National Organic Program, meaning that there was no blanket DCT, and minimal use of antimicrobials in general, with a loss of organic status of the animal if treated with antimicrobials. Breakpoints (where available) were used to categorize isolates as resistant, intermediate, or susceptible to antimicrobials. The MIC distributions of isolates from different herd types were compared using binomial or multinomial logistic regression. Of 240 CNS isolates, 12.9, 0.8, 7.1, 32.6, and 1.2%, were intermediate or resistant to ampicillin, cephalothin, erythromycin, penicillin, and tetracycline, respectively. Of 320 Staph. aureus isolates, 29.0, 2.5, 1.2, and 34.9% were intermediately resistant or resistant to ampicillin, penicillin, erythromycin, and oxacillin, respectively. Of 184 Strep. uberis isolates, 1.1, 25.0, 1.6, and 1.6% were intermediately resistant or resistant to erythromycin, penicillin, pirlimycin, and tetracycline, respectively. Generally, the MIC of CNS and streptococcal isolates from organic herds were lower than isolates from herds using DCT. However, the differences in MIC distributions occurred at MIC below clinical breakpoints, so that the bacteriological cure rates may not differ between isolates of differing MIC. Bimodal distributions of MIC for ampicillin and penicillin were found in Staph. aureus isolates from organic herds, suggesting that isolates with a higher MIC are a natural part of the bacterial population of the bovine mammary gland, or that isolates with higher MIC have persisted within these organic herds from a time when antimicrobials had been used. Given these observations, further work is required to determine if exposure to DCT is causally associated with the risk of elevated MIC, and whether reduction or removal of DCT from herds would reduce the risk of elevated MIC of mastitis pathogens.Item Longitudinal study of herd udder hygiene and its association with clinical mastitis in pasture-based dairy cows(Elsevier Inc and the Federation of Animal Science Societies (FASS) Inc on behalf of the American Dairy Science Association, 2021-05) Rowe S; Tranter W; Laven RThe objectives of this exploratory study were to (1) describe the association between herd-level udder hygiene and clinical mastitis and (2) investigate how sample size and milking stage affect the accuracy and precision of herd udder hygiene assessments made at milking time. A prospective longitudinal study was conducted in a dairy herd in Northern Australia as part of a previously published clinical trial of premilking teat disinfection. Video footage from 35 afternoon milkings was used to conduct 12,544 udder hygiene scores from 504 cows during an 89-d period and measure udder hygiene of the herd (proportion of cows with udder hygiene ≥3 out of 4). Linear interpolation was used to estimate herd udder hygiene on the days that were not scored, such that a herd-level udder hygiene measure was available for all cow-days in the study. Clinical mastitis events occurring during the study period were detected and recorded by farm staff according to a standardized definition. The relationship between herd udder hygiene on each of 1, 2, and 3 d before each study day (d -1, -2, and -3, respectively) and clinical mastitis at the cow level on each study day (each in turn being set as d 0) was determined using multivariable generalized estimating equations (family = Poisson, link = log), with the unit of analysis being the cow-day, adjusting for potential confounders and the clustering within the data. In addition, sampling strategies were evaluated by simulating herd udder hygiene assessments using a subset of cows in the herd. Herd udder hygiene from d -1, -2, and -3 was positively associated with clinical mastitis on d 0 (incidence rate ratio = 1.4 per 10-point increase in the percentage of cows with poor udder hygiene). Sampling strategy simulation found that at least 80 cows needed to be scored to achieve sufficiently precise estimations of herd udder hygiene. Furthermore, cows scored later during milking were slightly more likely to have poor udder hygiene than those scored earlier (risk ratio = 1.02 for cows that were 10% later in the milking order). More research is needed to evaluate risk factors for poor udder hygiene and potential interventions in pasture-based dairy cows.Item The Genetic Relatedness and Antimicrobial Resistance Patterns of Mastitis-Causing Staphylococcus aureus Strains Isolated from New Zealand Dairy Cattle(MDPI (Basel, Switzerland), 2021-11-22) Greening SS; Zhang J; Midwinter AC; Wilkinson DA; McDougall S; Gates MC; French NP; Butaye PStaphylococcus aureus is one of the leading causes of bovine mastitis worldwide and is a common indication for use of antimicrobials on dairy farms. This study aims to investigate the association between on-farm antimicrobial usage and the antimicrobial resistance (AMR) profiles of mastitis-causing S. aureus. Whole-genome sequencing was performed on 57 S. aureus isolates derived from cows with either clinical or subclinical mastitis from 17 dairy herds in New Zealand. The genetic relatedness between isolates was examined using the core single nucleotide polymorphism alignment whilst AMR and virulence genes were identified in-silico. The association between gene presence-absence and sequence type (ST), antimicrobial susceptibility and dry cow therapy treatment was investigated using Scoary. Altogether, eight STs were identified with 61.4% (35/57) belonging to ST-1. Furthermore, 14 AMR-associated genes and 76 virulence-associated genes were identified, with little genetic diversity between isolates belonging to the same ST. Several genes including merR1 which is thought to play a role in ciprofloxacin-resistance were found to be significantly overrepresented in isolates sampled from herds using ampicillin/cloxacillin dry cow therapy. Overall, the presence of resistance genes remains low and current antimicrobial usage patterns do not appear to be driving AMR in S. aureus associated with bovine mastitis.Item Therapy of Subclinical Mastitis during Lactation(MDPI (Basel, Switzerland), 2022-02-07) McDougall S; Clausen LM; Hussein HM; Compton CWR; Krömker VThis study tested the hypothesis that increasing the duration and/or frequency of antimicrobial treatment of subclinical mastitis would result in a higher bacteriological cure rate. Glands with a positive California mastitis test (CMT) from cows with an elevated somatic cell count (>500,000 cells/mL) that had an intramammary infection were randomly assigned at cow level to no treatment (Control; n = 80 glands), intramammary infusion of 200 mg cloxacillin sodium on three occasions at 48 h intervals (3 × 48 h; n = 273 glands), five occasions at 24 h intervals (5 × 24 h; n = 279 glands), or on five occasions at 48 h intervals (5 × 48 h; n = 72 glands). Glands were resampled at 21 (±3) and 28 (±3) days after initiation of treatment. The gland-level cure rate for any pathogen was 5/80 (6.2%), 139/173 (49.8%), 172/297 (61.6%) and 58/72 (80.6%) for Control, 3 × 48 h, 5 × 24 h and 5 × 48 h, respectively. The cure rate for major pathogens (defined as Staphylococcus aureus or Streptococcus spp.) was 4/52 (7.7%), 84/197 (42.6%), 96/183 (52.5%) and 36/48 (75%) for Control, 3 × 48 h, 5 × 24 h and 5 × 48 h, respectively. We conclude that treatment was superior to no treatment, and bacteriological cure rate was higher with the 5 × 24 h protocol than for the 3 × 48 h protocol and was higher with the 5 × 48 h than the 5 × 24 h protocol.