A scoping review of antimicrobial resistance in the Australian dairy cattle industry.

INTRODUCTION: Quantification of antimicrobial resistance (AMR) is beneficial to inform policies and direct prudent antimicrobial use. AIM: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Invited review: Selective treatment of clinical mastitis in dairy cattle.

Treatment of clinical mastitis (CM) and use of antimicrobials for dry cow therapy are responsible for the majority of animal-defined daily doses of antimicrobial use (AMU) on dairy farms. However, advancements made in the last decade have enabled excluding nonsevere CM cases from antimicrobial treatment that have a high probability of cure without antimicrobials (no bacterial causes or gram-negative, excluding Klebsiella spp.) and cases with a low bacteriological cure rate (chronic cases). These advancements include availability of rapid diagnostic tests and improved udder health management practices, which reduced the incidence and infection pressure of contagious CM pathogens. This review informed an evidence-based protocol for selective CM treatment decisions based on a combination of rapid diagnostic test results, review of somatic cell count and CM records, and elucidated consequences in terms of udder health, AMU, and farm economics. Relatively fast identification of the causative agent is the most important factor in selective CM treatment protocols. Many reported studies did not indicate detrimental udder health consequences (e.g., reduced clinical or bacteriological cures, increased somatic cell count, increased culling rate, or increased recurrence of CM later in lactation) after initiating selective CM treatment protocols using on-farm testing. The magnitude of AMU reduction following a selective CM treatment protocol implementation depended on the causal pathogen distribution and protocol characteristics. Uptake of selective treatment of nonsevere CM cases differs across regions and is dependent on management systems and adoption of udder health programs. No economic losses or animal welfare issues are expected when adopting a selective versus blanket CM treatment protocol. Therefore, selective CM treatment of nonsevere cases can be a practical tool to aid AMU reduction on dairy farms.

Selective treatment of nonsevere clinical mastitis does not adversely affect cure, somatic cell count, milk yield, recurrence, or culling: A systematic review and meta-analysis.

Treatment of clinical mastitis (CM) contributes to antimicrobial use on dairy farms. Selective treatment of CM based on bacterial diagnosis can reduce antimicrobial use, as not all cases of CM will benefit from antimicrobial treatment, e.g., mild and moderate gram-negative infections. However, impacts of selective CM treatment on udder health and culling are not fully understood. A systematic search identified 13 studies that compared selective versus blanket CM treatment protocols. Reported outcomes were synthesized with random-effects models and presented as risk ratios or mean differences. Selective CM treatment protocol was not inferior to blanket CM treatment protocol for the outcome bacteriological cure. Noninferiority margins could not be established for the outcomes clinical cure, new intramammary infection, somatic cell count, milk yield, recurrence, or culling. However, no differences were detected between selective and blanket CM treatment protocols using traditional analyses, apart from a not clinically relevant increase in interval from treatment to clinical cure (0.4 d) in the selective group and higher proportion of clinical cure at 14 d in the selective group. The latter occurred in studies co-administering nonsteroidal anti-inflammatories only in the selective group. Bias could not be ruled out in most studies due to suboptimal randomization, although this would likely only affect subjective outcomes such as clinical cure. Hence, findings were supported by a high or moderate certainty of evidence for all outcome measures except clinical cure. In conclusion, this review supported the assertion that a selective CM treatment protocol can be adopted without adversely influencing bacteriological and clinical cure, somatic cell count, milk yield, and incidence of recurrence or culling.

Udder health of dairy cows with an extended voluntary waiting period from calving until the first insemination.

This study aimed to evaluate the effect of an extended voluntary waiting period (VWP) on SCC, SCC elevations and clinical mastitis incidence during the complete lactation and the first 6 weeks of the next lactation. Holstein-Friesian dairy cows (N = 154) were blocked for parity, expected milk yield, calving season and breeding value for persistency and were randomly distributed across 3 VWP (50, 125, or 200 d: VWP-50, VWP-125, VWP-200). Cows were monitored from calving until 6 weeks into the next lactation, or until culling. An elevation of SCC in milk was defined as SCC in milk ≥200 000 cells/ml after two previous weeks with SCC < 200 000 cells/ml. Over the complete lactation, extending the VWP did not affect SCC elevations and the occurrence of clinical mastitis per lactation or per cow per year. There was no clear effect of VWP length on SCC in the complete lactation, except that multiparous cows in VWP-125 had a higher SCC compared with multiparous cows in VWP-50. Dry-off antibiotic usage per cow per year was lower in VWP-200 compared with VWP-50 for multiparous cows. In the first 6 weeks of the next lactation, cows in VWP-200 had a higher SCC compared with cows in VWP-50, with no effect of VWP on the number of elevations of SCC or the occurrence of clinical mastitis. Extending the VWP may therefore be used to reduce the frequency of transition periods and the associated use of dry-cow antibiotics, with limited impact on udder health, and a similar occurrence of SCC elevations and clinical mastitis per year.

Invited review: Selective use of antimicrobials in dairy cattle at drying-off.

Administering intramammary antimicrobials to all mammary quarters of dairy cows at drying-off [i.e., blanket dry cow therapy (BDCT)] has been a mainstay of mastitis prevention and control. However, as udder health has considerably improved over recent decades with reductions in intramammary infection prevalence at drying-off and the introduction of teat sealants, BDCT may no longer be necessary on all dairy farms, thereby supporting antimicrobial stewardship efforts. This narrative review summarizes available literature regarding current dry cow therapy practices and associated impacts of selective dry cow therapy (SDCT) on udder health, milk production, economics, antimicrobial use, and antimicrobial resistance. Various methods to identify infections at drying-off that could benefit from antimicrobial treatment are described for selecting cows or mammary quarters for treatment, including utilizing somatic cell count thresholds, pathogen identification, previous clinical mastitis history, or a combination of criteria. Selection methods may be enacted at the herd, cow, or quarter levels. Producers' and veterinarians' motivations for antimicrobial use are discussed. Based on review findings, SDCT can be adopted without negative consequences for udder health and milk production, and concurrent teat sealant use is recommended, especially in udder quarters receiving no intramammary antimicrobials. Furthermore, herd selection should be considered for SDCT implementation in addition to cow or quarter selection, as BDCT may still be temporarily necessary in some herds for optimal mastitis control. Costs and benefits of SDCT vary among herds, whereas impacts on antimicrobial resistance remain unclear. In summary, SDCT is a viable management option for maintaining udder health and milk production while improving antimicrobial stewardship in the dairy industry.

Consequences of extending the voluntary waiting period for insemination on reproductive performance in dairy cows.

The aim of the study was to evaluate the effect of extended voluntary waiting period (VWP) on ovarian cyclicity and reproductive performance of dairy cows. Holstein-Friesian dairy cows (N = 154) were blocked and randomly assigned to one of 3 groups with different VWP (50, 125 or 200 d: VWP-50, VWP-125 or VWP-200). Milk samples were collected 3 times a week and analysed for progesterone concentration. Ovarian cycles were classified as: normal (18-24 days), short (<18 days) or prolonged (>24 days). For cows that became pregnant within 100 days after VWP, a VWP-200 d was related with fewer days until pregnancy after end of the VWP (19.4 d) compared with VWP-50 or VWP-125 (35.5, 37.3 d respectively). During 100 days (-50 until 50 d) around the end of VWP, cows in VWP-200 had a greater percentage of normal cycles (91.9 vs 58.0 %, P < 0.01) and a lower percentage of prolonged cycles (6.0 vs 32.7 %, P = 0.01) compared with cows in VWP-50. In the 4 weeks around the end of the VWP, cows in VWP-125 and VWP-200 had a lower milk yield compared with cows in VWP-50 (32.0, 27.5 vs 37.4 kg/d, P < 0.01). Inseminations continued until 300 days in milk, resulting in fewer pregnant cows for longer VWPs. In conclusion, extending the VWP from 50 to 125 or 200 days resulted in a greater percentage of cows with normal ovarian cycles and a lower milk yield around the end of VWP. Moreover, VWP-200 reduced days open after the end of the VWP, compared with VWP-50.

Antimicrobial use on Australian dairy cattle farms - A survey of veterinarians.

AIMS: The aims of this study were to determine antimicrobial prescription patterns and the factors affecting antimicrobial selection amongst Australian dairy veterinarians. METHODS: A structured questionnaire was administered to Australian dairy cattle veterinarians using the Qualtrics online survey platform. Questions focused on their (1) demographics; (2) opinions surrounding antimicrobial use, resistance, and stewardship; (3) decision-making drivers of both prescription and selection of commonly prescribed antimicrobials; (4) awareness on the guidelines for antimicrobial usage and sources of information concerning antimicrobials. KEY RESULTS: A total of 135 responses (14.1% response rate) from all eight dairying regions in Australia were received. The attitudes, perceptions, and concerns of dairy veterinarians towards antimicrobials indicated a high agreement regarding label indications (96%), consequences of off-label prescription (95%), and the presence of an antimicrobial resistance (AMR) risk (73%), when prescribing antibiotics. A four-dimensional categorical principal components analysis (CATPCA) model indicated most of the variation in opinion was due to AMR risk, trade-offs, prescription concerns and active substance concerns. The first active substance most dairy veterinarians chose for a scenario involving mastitis and dry cow therapy (DCT) treatment was cloxacillin. Decision-making drivers for antimicrobial choice when providing advice regarding the supply of antimicrobials for mastitis and DCT treatment were predominately clinical factors; however, diagnostics were rarely used in determining antimicrobial choice due to cost of implementation, diagnostic accuracy (sensitivity, specificity), and benefit issues. Non-clinical decision-making drivers included the perception of practicality for Australian Veterinary Association (AVA) prescription guidelines, opinions surrounding AMR risk and prescription concerns, consideration of Expert Advisory Group on Antimicrobial Resistance (EAGAR) scores, number of years worked with dairy farms, and the number of dairy farms they regularly consult for. When available at the practice, prescription policies were considered to impact on animal welfare outcomes and on the probability of AMR emergence. The major information sources influencing decision making on antimicrobial prescription for the Australian dairy veterinarians were clinical experience (93%) and product labels (81%). CONCLUSIONS: Australian dairy veterinarians are generally aware of the risk of resistance to antimicrobials and the need for stewardship, with clinical factors having the most impact on antimicrobial prescription. However, non-clinical factors incorporating awareness of guidelines and their attitudes on antimicrobial resistance risk and prescription concerns impact on the choice and prescription of antimicrobials. IMPLICATIONS: The development of prescription policy and guidelines, alongside effective communicative extension programs to increase veterinarian uptake, provides an avenue to mitigate AMR risk in Australian dairy cattle.

A stochastic modelling approach to determine the effect of diverse Staphylococcus aureus strains on the economic and epidemiological outcomes of mastitis intervention strategies in dairy cattle.

Staphylococcus aureus (S. aureus) strains with considerable genetic and phenotypic differences have previously been identified. The economic and epidemiologic impact of S. aureus mastitis has been investigated, but none of these studies took differences between strains into account. Here we aimed to investigate how differences between S. aureus strains affect the economic and epidemiologic outcome of various intervention strategies against clinical and subclinical intramammary infections. Five S. aureus strains were modelled using a stochastic bio-economic model simulating a dairy herd of 200 cows using single-day time steps. The strain characteristics of the five simulated S. aureus strains (general, contagious, spill-over, clinical and persistent) were based on divergent phenotypes as described in literature. Outcomes of the model included incidence (both clinical and subclinical), number of antibiotic treatment days, number of culled cows, and net income. Intervention strategies against clinical and subclinical intramammary infections were based on (variations of) intramammary antibiotic treatment, testing, and culling. Both single and multiple pathogen (intramammary infection caused by S. aureus, Escherichia coli, and non-aureus staphylococci) scenarios were simulated to determine the effect of the five S. aureus strains on the impact of 19 different intervention strategies. The results showed that the incidence (both clinical and subclinical), number of treatment days, number of culled cows, and net income varied considerably for the different S. aureus strains. Comparison of the model outcomes within and between strains showed that for most intervention strategies the relative impact differed per strain. However, the intervention strategy with the best outcome for most variables and strains was the culling of cows with a recovery probability lower than 50%. This shows that the relative economic and epidemiologic impact of most of the modelled intervention strategies were strain-dependent, while some intervention strategies were not strain-dependent. From this, we conclude that, depending on the intervention strategy applied on a farm, it could be advantageous to type S. aureus to determine whether it would be economically and epidemiologically beneficial for the existing intervention strategy to be changed.

Transmission dynamics of Staphylococcus aureus and Streptococcus agalactiae in a Dutch dairy herd using an automatic milking system.

Staphylococcus aureus and Streptococcus agalactiae are important contagious mastitis pathogens and are considered to mainly transmit between cows through the milking machine. Controlling contagious mastitis on dairy farms requires a reduction of the transmission rate or the duration of intramammary infections (IMI), or both. These parameters may differ in dairy herds milked with an automatic milking system (AMS) as compared to those milked with a conventional milking system (CMS). The aims of this prospective longitudinal study were to estimate the transmission rate, the median duration of IMI and the basic reproduction number (R0) of Staph. aureus and Strep. agalactiae in a Dutch AMS herd. Bacteriological cultures of quarter milk samples were collected every 2 wks. Using 3 different definitions of IMI, we estimated the transmission rate for Staph. aureus to be within the range of 0.002 (95 % CI: 0-0.005) quarter-day-1 to 0.019 (95 % CI: 0.010-0.032) quarter-day-1, and for Strep. agalactiae of 0.007 (95 % CI: 0.005-0.010) quarter-day-1 to 0.019 (95 % CI: 0.011-0.032) quarter-day-1, the median duration of chronic IMI at 95 (95 % CI: 72-125) days for Staph. aureus and at 86 (95 % CI: 67-111) days for Strep. agalactiae, and the R0 between 0.16 (95 % CI: 0.05-0.27) and 0.34 (95 % CI: 0.20-0.48) for Staph. aureus, and between 0.64 (95 % CI: 0.41-0.87) and 0.68 (95 % CI: 0.48-0.88) for Strep. agalactiae. Transmission of these two contagious pathogens in this herd was limited and theoretically the IMI would not sustain, given that R0 of both pathogens was lower than 1. The estimated transmission rate of Staph. aureus in this AMS herd was found to be comparable to those described for CMS herds, while for Strep. agalactiae, it was slightly higher than in CMS herds. The duration of Staph. aureus IMI was in line with results from CMS farms, while the duration of Strep. agalactiae was lower than what has been described in CMS herds. The R0 of these contagious pathogens was found to be lower than the estimates in CMS herds. Our study suggests that the transmission rate of these two contagious pathogens in this AMS herd were comparable to what has been reported about well-performing CMS herds that have a low rate of transmission.

Effects of a mastitis treatment strategy with or without on-farm testing.

The etiology of mastitis is crucial information to use antimicrobials prudently for control and treatment. This study aimed to evaluate the effects of mastitis diagnosis and treatment strategies with on-farm testing, on cure, new intramammary infections (IMI), somatic cell count (SCC), and antimicrobial use, compared with farmers' current diagnosis and treatment strategies. The on-farm tests used, CHROMagar Mastitis (CHROMagar, Paris, France) and Minnesota Easy Culture System II Tri-plate (University of Minnesota, St. Paul, MN), both had etiological groups of IMI as result, being gram-positive growth, gram-negative growth, or culture negative. Two randomized controlled trials were conducted on 15 herds: trial 1 prospectively enrolled 155 cows with clinical mastitis, and trial 2 cross-sectionally included 78 cows with subclinical mastitis. In both trials, cows were randomly distributed over 3 equal-sized groups: a test group using CHROMagar, a test group using Minnesota, and a control group not using on-farm tests. Farmers decided whether or not to treat, and which antimicrobial treatment would be applied, using information available on the day of enrollment (control group), complemented with the on-farm test result 1 d after enrollment (both test groups). For clinical mastitis, an antimicrobial treatment was given in 58% of cases that used CHROMagar, in 80% that used Minnesota, and in 86% of the controls. For subclinical mastitis, an antimicrobial treatment was given in 50% of cases that used CHROMagar, in 54% that used Minnesota, and in 4% of the controls. Bacteriological cure rate of clinical mastitis was lowest in the CHROMagar group [odds ratio 0.18 (95%CI 0.03-0.99)] compared with the controls. Using the Minnesota on-farm test for subclinical mastitis diagnosis and treatments resulted in fewer new IMI on d 21 [odds ratio 0.06 (95%CI 0.00-0.74)] compared with the controls. Clinical cure rate, percentage of new IMI, and SCC on d 21 of clinical mastitis were comparable among the groups. Using on-farm tests in farmers' decision-making process resulted in more treatments in accordance with the etiology of mastitis than without on-farm testing. A diagnosis and treatment strategy with on-farm testing is advised in cows with clinical mastitis to enhance prudent antimicrobial use. For subclinical mastitis, however, on-farm testing may lead to an unacceptable increase in use of antimicrobials and thus should not be advised as the common approach.

Dynamics of intramammary infections in suckler ewes during early lactation.

Mastitis is an important problem in meat-producing sheep, but few studies have investigated the transmission dynamics of mastitis pathogens in these animals. The objective of this study was to describe the pathogens causing intramammary infections (IMI) in suckler ewes, their effect on somatic cell count, and the dynamics of these IMI in early lactation. We enrolled 15 sheep flocks early after lambing and selected ewes in each flock that were sampled twice with a 3-wk interval. Milk samples from both glands of each ewe were bacteriologically cultured, and somatic cell count was measured. Non-aureus Staphylococcus spp. were the most prevalent culture results. Somatic cell counts were most strongly increased in ewes infected with Mannheimia haemolytica, whereas staphylococci, including Staphylococcus aureus, were associated with a moderate increase in somatic cell count. The proportion of udder halves that remained culture-positive with Staphylococcus spp. during the 3-wk sampling interval was moderate, but M. haemolytica infections were stable during this time period. A substantial number of new infections were seen in the early lactation study period for non-aureus Staphylococcus spp., Staph. aureus, and Corynebacterium spp., but not for M. haemolytica or Streptococcus spp. The number of new IMI of Staph. aureus was associated with the number of Staph. aureus-infected udder halves in the flock at the first sampling moment, indicative of contagious transmission. Altogether, we show that substantial transmission happens in early lactation in suckler ewes, but that the dynamics differ between pathogen species. More research is needed to further describe transmission in different stages of lactation and to identify transmission routes, to develop effective interventions to control mastitis.

Genomic analysis of European bovine Staphylococcus aureus from clinical versus subclinical mastitis.

Intramammary infections (IMI) with Staphylococcus aureus are a common cause of bovine mastitis and can result in both clinical (CM) or subclinical mastitis (SCM). Although bacterial isolates of S. aureus differ in their virulence potential it is largely unclear which bacterial virulence factors are responsible for increased clinical severity. We performed a genome wide association study and used a generalized linear mixed model to investigate the correlation between gene carriage, lineage and clinical outcome of IMI in a collection of S. aureus isolates from cattle with CM (n = 125) and SCM (n = 151) from 11 European countries. An additional aim was to describe the genetic variation of bovine S. aureus in Europa. The dominant lineages in our collection were clonal complex (CC) 151 (81/276, 29.3%), CC97 (54/276, 19.6%), CC479 (32/276, 11.6%) and CC398 (19/276, 6.9%). Virulence and antimicrobial resistance (AMR) gene carriage was highly associated with CC. Among a selection of nine virulence and AMR genes, CC151, CC479 and CC133 carried more virulence genes than other CCs, and CC398 was associated with AMR gene carriage. Whereas CC151, CC97 were widespread in Europe, CC479, CC398 and CC8 were only found in specific countries. Compared to CC151, CC479 was associated with CM rather than SCM (OR 3.62; 95% CI 1.38-9.50) and the other CCs were not. Multiple genes were associated with CM, but due to the clustering within CC of carriage of these genes, it was not possible to differentiate between the effect of gene carriage and CC on clinical outcome of IMI. Nevertheless, this study demonstrates that characterization of S. aureus CC and virulence genes helps to predict the likelihood of the occurrence of CM following S. aureus IMI and highlights the potential benefit of diagnostics tools to identify S. aureus CC during bovine mastitis.

Consequences of Transition Treatments on Fertility and Associated Metabolic Status for Dairy Cows in Early Lactation.

This study aimed to (1) investigate effects of reducing postpartum dietary energy level for cows after a 0-d dry period (DP) on resumption of ovarian cyclicity and reproductive performance, (2) relate days open with other reproductive measures, and (3) relate onset of luteal activity (OLA) and days open with metabolic status in early lactation. Holstein-Friesian dairy cows were randomly assigned to 1 of 3 transition treatments: no DP and low postpartum dietary energy level from 22 days in milk( DIM )onwards (0-d DP (LOW)) (n = 42), no DP and standard postpartum dietary energy level (0-d DP (STD)) (n = 43), and a short DP and standard postpartum dietary energy level (30-d DP (STD)) (n = 43). Milk progesterone concentration was determined three times per week until 100 DIM. Plasma metabolite and hormone concentrations were measured weekly until week 7 postpartum. Reducing postpartum dietary energy level in older cows (parity ≥ 3) after no DP and 22 DIM did not affect milk production but prevented a positive energy balance and shortened the interval from calving to OLA. In addition, services per pregnancy and days open were reduced in cows of parity ≥ 3 on 0-d DP (LOW), compared with cows of parity ≥ 3 with 0-d DP (STD), but not in cows of parity 2.

Performance of Online Somatic Cell Count Estimation in Automatic Milking Systems.

Somatic cell count (SCC) is one of the most important and widely used mastitis diagnostics. For detecting (sub)clinical mastitis, online SCC related measurements are more and more used in automatic milking systems (AMS). Sensors such as an automated online California Mastitis Test (O-CMT) allow for high frequency screening of high SCC cows within a herd, which makes it potentially powerful to identify episodes of mastitis. However, the performance of O-CMT measurements, as compared to SCC determined in the laboratory (L-SCC), has only scarcely been described. The aims of this study were (1) to assess the agreement between the O-CMT measurement averaged over different time windows and the corresponding L-SCC measurements; (2) to determine the optimal time window for averaging O-CMT as compared to L-SCC; (3) to explore the added value of time-series of frequent O-CMT measurements in individual cow udder health monitoring compared to L-SCC measurements. Data were collected from 50 farms in 6 different countries that were equipped with AMS using O-CMT measurements and also performed regular L-SCC testing. We found that the overall concordance correlation coefficient (CCC) between O-CMT and L-SCC was 0.53 but differed substantially between farms. The CCC between O-CMT and L-SCC improved when averaging O-CMT over multiple milkings, with an optimal time-window of 24 h. Exploration of time series of daily O-CMT recordings show that this is an effective screening tool to find episodes of high SCC. Altogether, we conclude that although O-CMT agrees moderately with L-SCC, because of its high measurement frequency, it is a promising on-farm tool for udder health monitoring.

Knowledge, attitude and practices of Swiss dairy farmers towards intramammary antimicrobial use and antimicrobial resistance: A latent class analysis.

Understanding farmers' mindsets is important to improve antimicrobial stewardship in the dairy industry. This cross-sectional study aimed to determine farmers' knowledge, attitude, and practices with respect to lactational intramammary antimicrobial use (AMU) and antimicrobial resistance (AMR) in Swiss dairy herds. Based on their approach towards subclinical mastitis (SCM) and non-severe cases of clinical mastitis (CM), subgroups of farmers were identified and compared regarding their knowledge, attitude and practices towards AMU and AMR. After conducting qualitative interviews to develop a questionnaire, an online survey was sent to 1296 randomly selected Swiss dairy farmers. Information was gathered on demographic data and farmers' knowledge, attitude, and practices towards AMU and AMR. A latent class analysis was performed to identify subgroups of farmers based on management of SCM and non-severe CM cases. Based on the results of 542 completed questionnaires, poor knowledge with respect to AMU and AMR was identified, as well as discrepancies between farmers' perceptions and their actual practices. Farmers approached cows with SCM and non-severe CM similarly, indicating they perceived both mastitis states as the same disease. Intramammary antimicrobial products containing cefquinome, which is a highest priority critically important antimicrobial, were among the 3 most commonly applied intramammary antimicrobials. Five latent classes of farmers were identified based on their management towards SCM and non-severe CM. One group of farmers (18.5% of respondents) indicated that they did not treat those mastitis cases, one group only treated SCM cases (13.8% of respondents), one group only treated non-severe CM cases (28.6% of respondents) and the largest group treated both mastitis states (39.1% of respondents). The latter group was subdivided into a latent class of farmers following guidelines for AMU/AMR (25.5% of respondents) and a group of farmers that were not strictly following these guidelines (13.7% of respondents). Regional differences between farmers, according to altitude and language region, explained some of the variation in latent class membership. Latent class membership was associated with farmers' attitude to use antimicrobials as little as possible and with using antimicrobials only after performing bacteriological and susceptibility testing. This study gave detailed insight into Swiss farmers' knowledge, attitude, and practices regarding AMU and AMR and provides opportunities to improve antimicrobial stewardship in Swiss dairy herds. The identified groups of farmers, based on their management practices regarding SCM and non-severe CM, may help to design tailored intervention strategies for improving prudent AMU in the heterogeneous population of dairy farmers in Switzerland.

Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent.

Bovine mastitis is a costly disease to the dairy industry and intramammary infections (IMI) with Staphylococcus aureus are a major cause of mastitis. Staphylococcus aureus strains responsible for mastitis in cattle predominantly belong to ruminant-associated clonal complexes (CCs). Recognition of pathogens by bovine mammary epithelial cells (bMEC) plays a key role in activation of immune responsiveness during IMI. However, it is still largely unknown to what extent the bMEC response differs according to S. aureus CC. The aim of this study was to determine whether ruminant-associated S. aureus CCs differentially activate bMEC. For this purpose, the immortalized bMEC line PS was stimulated with S. aureus mastitis isolates belonging to four different clonal complexes (CCs; CC133, CC479, CC151 and CC425) and interleukin 8 (IL-8) release was measured as indicator of activation. To validate our bMEC model, we first stimulated PS cells with genetically modified S. aureus strains lacking (protein A, wall teichoic acid (WTA) synthesis) or expressing (capsular polysaccharide (CP) type 5 or type 8) factors expected to affect S. aureus recognition by bMEC. The absence of functional WTA synthesis increased IL-8 release by bMEC in response to bacterial stimulation compared to wildtype. In addition, bMEC released more IL-8 after stimulation with S. aureus expressing CP type 5 compared to CP type 8 or a strain lacking CP expression. Among the S. aureus lineages, isolates belonging to CC133 induced a significantly stronger IL-8 release from bMEC than isolates from the other CCs, and the IL-8 response to CC479 was higher compared to CC151 and CC425. Transcription levels of IL-8, tumor necrosis factor alpha (TNFα), serum amyloid A3 (SAA3), Toll-like receptor (TLR)-2 and nuclear factor κB (NF-κB) in bMEC after bacterial stimulation tended to follow a similar pattern as IL-8 release, but there were no significant differences between the CCs. This study demonstrates a differential activation of bMEC by ruminant-associated CCs of S. aureus, which may have implications for the severity of mastitis during IMI by S. aureus belonging to these lineages.

Prevalence of extended-spectrum and AmpC ß-lactamase-producing Escherichia coli in Dutch dairy herds.

This study was conducted to assess: (1) a change in between-herd prevalence of extended-spectrum and AmpC ß-lactamase-producing Escherichia coli (ESBL/AmpC-EC) between 2011 and 2013, the period during which the antimicrobial policy in animal husbandry in the Netherlands changed significantly, and (2) the prevalence of ESBL/AmpC-EC in individual calves, young stock, and dairy cows in the Netherlands. In 196 randomly selected conventional dairy herds, faecal samples were collected from calves (maximum n = 15), and randomly selected young stock (n = 5) and dairy cows (n = 15). Additionally, fresh faecal samples were collected from five different places on the floors where the dairy cows were housed. Samples were screened for E. coli with non-wild type susceptibility for cefotaxime and isolates were phenotypically confirmed as ESBL/AmpC-producing by disc diffusion, using cefotaxime and ceftazidime with and without clavulanic acid, and cefoxitin. Samples containing ESBL/AmpC-EC were examined semi-quantitatively. In 59.6% of the dairy herds one or more samples tested positive for ESBL/AmpC-EC. The between-herd prevalence based on floor samples in 2013 (18.0%) was significantly lower than the prevalence in 2011 based on comparable samples (32.7%). The individual animal prevalence of ESBL/AmpC-EC, with a minimum shedding level of 103 cfu/g of faeces, was 19.3% in calves, 0.9% in young stock, and 0.8% in dairy cows. Although ESBL/AmpC-EC was found in the majority of dairy herds, the herd prevalence declined significantly between 2011 and 2013. Calves were found to have both, a much higher individual animal prevalence and a higher level of shedding than young stock and cows.

Intramammary antimicrobial treatment of subclinical mastitis and cow performance later in lactation.

The aim of this study was to evaluate long-term therapeutic effects of antimicrobial treatment of recently acquired subclinical mastitis (RASCM) during lactation. Quarter-level clinical mastitis (CM) follow-up, composite somatic cell counts (SCC), and cow-level milk yield later in lactation were evaluated using follow-up data from 2 previously published linked randomized field trials. The first trial randomly assigned antimicrobial treatment with any intramammary product or negative control to culture-positive quarters of cows having a first elevated composite SCC after 2 consecutive low composite SCC measurements. Untreated cows that had a second elevated composite SCC at the next measurement and were staphylococci-positive (i.e., Staphylococcus aureus or non-aureus staphylococci) were randomly assigned to treatment or control. Quarter-level CM cases were reported by the participating herd personnel, and milk yield and composite SCC data were obtained from the regular test-day recording. Frailty survival models were used to evaluate the long-term therapeutic effects of antimicrobial treatment of RASCM on quarter-level CM follow-up. Mixed linear regression models were applied to quantify the effect on milk yield and composite SCC. Data of 638 quarters from 486 cows in 38 herds were available for statistical analyses, of which 229 quarters of 175 cows received antimicrobial treatment for RASCM. Antimicrobial treatment culminated in reduced composite SCC levels later in lactation but did not result in different milk yield levels or CM follow-up compared with control cows. Antimicrobial treatment of cows with RASCM should therefore only be considered in exceptional situations given the current focus on antimicrobial usage reduction in animal husbandry.

Agreement between four commercial diagnostic tests and routine bacteriological culture of milk to determine the udder infection status of dairy cows.

Mastitis is usually treated based on clinical signs or somatic cell count information rather than on results of bacteriological culture of milk. In many countries an optimal mastitis treatment is considered important from the perspective of therapy efficacy, prudent antimicrobial use and farm economics. Farmers can optimize their mastitis treatment decisions if they know whether and which mastitis pathogen is involved. Information on the mastitis pathogen involved can be acquired from diagnostic mastitis tests such as culture-based tests. This study aimed to determine the agreement of four commercial culture-based mastitis tests with routine bacteriological culture of milk to determine the intramammary infection status of a quarter or cow. The commercial culture-based tests evaluated in this study were CHROMagar Mastitis (CHROMagar, France), Hardy Diagnostics Mastitis Triplate (Hardy Diagnostics, USA), Minnesota Easy Culture System II Tri-plate (University of Minnesota, USA), and VétoRapid (Vetoquinol, the Netherlands). We used 866 prospectively collected milk samples, routinely submitted to the bacteriological laboratory of GD Animal Health for routine bacteriological culture of milk from April to June 2016. Samples were cultured on routine bacteriological culture of milk and on the commercial culture-based tests. We calculated the agreement beyond chance of each commercial culture-based test result with the result of routine bacteriological culture using 2x2 contingency tables. Furthermore, inter-reader agreement was determined for 597 samples read by two masked readers. The agreement of the four commercial culture-based mastitis tests with routine bacteriological culture of milk for Gram-positive bacteria ranged from 0.14 (95% CI 0.11-0.16) using Hardy Diagnostics Mastitis Triplate to 0.25 (95% CI 0.22-0.28) using Minnesota Easy Culture System II Tri-plate. The agreement for Gram-negative bacteria was approximately 0.70 (95% CI 0.66-0.74) for all four commercial culture-based tests. The agreement for no growth ranged from 0.22 (95% CI 0.19-0.25) using Hardy Diagnostics Mastitis Triplate to 0.34 (95% CI 0.31-0.38) using VétoRapid. This category was affected by prevalence and bias as the prevalence adjusted and bias adjusted kappa ranged from 0.63 (95% CI 0.56-0.69) using CHROMagar Mastitis to 0.68 (95% CI 0.62-0.74) using Hardy Diagnostic Mastitis Triplate. Agreement between readers was almost perfect. Although only for Gram-negative bacteria a good agreement was found between commercial culture-based tests and routine bacteriological culture of milk, and further on-farm evaluations are needed to determine the effect of these findings on udder health, commercial culture-based tests are of added value to support decisions whether and how to treat cows with mastitis.