Antimicrobial resistance of bacteria isolated from dairy cow milk samples submitted for bacterial culture: 8,905 samples (1994-2001)

OBJECTIVE: To determine whether antimicrobial resistance patterns of major mastitis pathogens isolated from milk samples from dairy cows have changed over time. DESIGN: Retrospective study. SAMPLE POPULATION: 8905 bacterial isolates obtained from milk samples submitted to the Wisconsin Veterinary Diagnostic Laboratory between January 1994 and June 2001. PROCEDURE: Antimicrobial susceptibility was determined by means of the Kirby-Bauer disk diffusion method. Logistic regression was used to determine whether percentages of isolates resistant to various antimicrobials changed over time. RESULTS: For the gram-positive mastitis pathogens, percentages of isolates resistant to various flactam antimicrobials did not increase over the course of the study. Percentage of Staphylococcus aureus isolates resistant to penicillin decreased from 49 to 30%; percentage of Streptococcus isolates resistant to penicillin decreased from 6 to 1%. Percentage of isolates resistant to erythromycin increased for S aureus, Escherichia coli, Enterobacter spp, Enterococcus spp, and Pasteurella spp. Percentage of isolates resistant to lincomycin increased for S aureus and Staphylococcus spp. Percentage of coagulase-negative Staphylococcus isolates resistant to pirlimycin increased from 6 to 19%. For several pathogens, percentages of isolates resistant to sulfisoxazole and to trimethoprim-sulfamethoxazole decreased. No pathogens had a significant increase in the percentage of isolates resistant to novobiocin-penicillin. CONCLUSIONS AND CLINICAL RELEVANCE: Results did not indicate a trend toward increased antimicrobial resistance among mastitis pathogens isolated from milk samples from dairy cows between 1994 and 2001. Reduced resistance to flactam antimicrobials was identified for several gram-positive mastitis pathogens.     

Culture results from milk samples submitted to veterinary diagnostic laboratories from August 2003 to December 2006 in New Zealand

Abstract

AIMS: To determine the pattern of isolation of major mastitis-causing organisms isolated from milk samples submitted to five veterinary diagnostic laboratories in New Zealand.

METHODS: The culture results of 25,288 milk samples that were collected from dairy cows throughout New Zealand from August 2003 to December 2006 and submitted to a group of veterinary diagnostic laboratories were assembled, reviewed and summarised. Logistic regression was used to analyse the effect of year, region (i.e. North vs South Island), and season on the probability of isolating the two most common organisms.

RESULTS: The most commonly isolated mastitis causing organisms from all samples were: Streptococcus uberis (23.6%), Staphylococcus aureus (23.5%), coagulase-negative staphylococci (CNS; 7.2%), Strep. dysgalactiae (6.2%), Bacillus spp. (4.0%), and coliforms (3.7%). The percentage of samples with isolates of Strep. uberis or Staph. aureus was affected by island, year and season (p<0.001). For most of the year, except in late winter and early spring when Strep. uberis was much more common, the percentage of isolates of Strep. uberis and Staph. aureus were not apparently different despite the former being an environmental pathogen and the other a contagious one.

CONCLUSION: The pattern of isolation of major mastitis-causing organisms, as determined from culture of milk samples submitted to diagnostic laboratories in New Zealand, has changed significantly over the last 40 years, with a substantial increase in the percentage of isolates that are Strep. uberis and a decrease in isolates of Strep. agalactiae. There is a clear seasonal pattern to the isolation of both Strep. uberis and Staph. aureus, particularly the former.

CLINICAL RELEVANCE: Knowledge of the aetiological agents causing bovine mastitis on a farm is of value in determining the choice of treatment. This dataset shows that, although there is seasonal pattern to the isolation of mastitis-causing organisms in New Zealand, both Strep. uberis and Staph. aureus are isolated throughout the year, so bacteriology is of value in determining aetiology even in late winter/early spring.     

Low levels of antibacterial drug resistance expressed by Gram-negative bacteria isolated from poultry carcasses in New Zealand

Abstract

AIM: To provide baseline data on the levels and patterns of antibacterial drug resistance expressed by Gram-negative bacteria isolated from poultry carcasses in New Zealand.

METHODS: Between July and December 2006, isolates of Escherichia coli (n=407) and Salmonella spp. (n=3) originating from carcass-rinse samples were submitted by testing laboratories affiliated to five major poultry processing plants. Isolates of Campylobacter jejuni (n=193) originating from retail poultry carcasses in 2005–2006 were retrieved from the Massey University archives. All isolates underwent disc diffusion susceptibility testing against panels of 12 (Enterobacteriaceae) and six (Campylobacter spp.) antibacterial drugs. Cephalothin-resistance in isolates of E. coli was confirmed using ETest strips, and confirmation of the resistance phenotypes for a subset of C. jejuni isolates used microbroth dilution assays. Patterns within the resistance phenotypes of the isolates were investigated using hierarchical clustering, and logistic regression modelling.

RESULTS: The majority of isolates (71.5% E. coli, 99% C. jejuni, and all three Salmonella spp. isolates) were fully susceptible to the drugs that were tested. Four (1%) E. coli isolates showed resistance to three or more drugs. The proportions of susceptible E. coli differed between the five processing plants. Resistances were detected in E. coli isolates, using disc diffusion to cephalothin (18.2%), ampicillin (4.4%), tetracycline (4.4%) and gentamicin (1.5%). There was an association between cephalothin-resistant isolates of E. coli and decreased susceptibility to gentamicin. Using ETests to ascertain the minimum inhibitory concentrations (MIC) of E. coli for cephalothin gave inconsistent results. One of 193 C. jejuni isolates was resistant to erythromycin, and microbroth dilution assays confirmed that this panel of C. jejuni was generally susceptible to antibacterial drugs.

CONCLUSIONS: The levels of resistance shown by Gram-negative bacteria isolated from chicken carcasses in New Zealand are among the lowest reported around the world. No resistance to extended-spectrum cephalosporin drugs was detected in E. coli, suggesting that CTX-M and AmpC beta-lactamases are rare or absent. Salmonella spp. are rarely isolated from poultry carcasses during routine testing in New Zealand, and the isolates identified during this study were fully susceptible to the drugs tested. A panel of C. jejuni isolates originating from retail poultry carcasses were susceptible to first-line and second-line antibacterial drugs. The use of cephalothin as a marker of resistance to first-generation cephalosporins may not be appropriate for non-type-specific E. coli of animal origin.     

兽医临床凝固酶阴性葡萄球菌分离株的耐药性及耐药基因检测

采用微量肉汤稀释法和D-试验法检测64株凝固酶阴性葡萄球菌（CNs）对10种抗菌药物的耐药性,并用PCR方法分别检测头孢西丁耐药菌株和红霉素耐药菌株中mecA基因以及erinA、ermB、ermC和msrA基因的携带情况。结果表明,所有菌株均对万古霉素和甲氧苄啶／磺胺甲恶唑敏感;泰妙菌素为耐药率（65．6％）最高的抗菌药物,其次是红霉素、氧氟沙星和β-内酰胺类药物。28株（43．8％）CNS对青霉素耐药,其中26株对头孢西丁耐药（MRS）并且均携带mecA基因。30株（46．9％）CNS对红霉素耐药,其中24株为MLSB耐药表型,主要由ermB基因介导。总之,兽医临床CNs分离株对常用抗菌药物的耐药性不同,mecA基因和ermB基因的携带分别是兽医临床MRS和MLSB表型产生的主要原因。     

Successful treatment of infectious (Salmonella type III: 44) polyarthritis and osteomyelitis in a 4‐week‐old foal

This article describes the acute onset of infectious polyarthritis and osteomyelitis in a 4‐week‐old foal. Analysis of synovial fluid obtained from the left femoropatellar and right tarsocrural joints combined with clinical signs consisting of joint effusion and lameness yielded a diagnosis of septic arthritis. Bacterial culture of synovial fluid from the left stifle revealed Salmonella type III: 44. Rapid, sustained clinical improvement was noted following discontinuation of empirical antimicrobial therapy (potassium penicillin and amikacin sulphate) and initiation of treatment with ceftiofur and ampicillin. The importance of combining knowledge of veterinary pharmacology and microbiology so that appropriate antimicrobials may be selected with regard to the local environment in which they are to eradicate infection is emphasised. Despite frequent reference to amikacin sulphate as an effective antimicrobial for treating infections in foals caused by Salmonella, factors are discussed that explain why amikacin may not be clinically effective for treating infectious arthritis caused by Salmonella.     

新疆伊犁某牛场大肠杆菌耐药性调查

为了调查新疆伊犁某牛场饮水、饲料和粪样中分离的大肠杆菌对临床常用抗菌药物的耐药情况。本试验采用微量肉汤稀释法,对饮水源、饲料源、牛粪源样品中分离出的大肠杆菌进行最小抑菌浓度测定。结果显示,25份牛场饮水源样品,大肠杆菌分离率为100.0%（25/25）,对阿莫西林/克拉维酸（12.0%）、氨苄西林（4.0%）、诺氟沙星（4.0%）、恩诺沙星（8.0%）和安普霉素（8.0%）5种抗菌药物耐药;72份牛场饲料源样品,大肠杆菌分离率为65.3%（47/72）,对阿莫西林/克拉维酸（36.2%）、氨苄西林（19.1%）、诺氟沙星（4.3%）和安普霉素（4.3%）4种抗菌药物耐药;80份牛粪源样品,大肠杆菌分离率为100.0%（80/80）,对阿米卡星（12.5%）、氨苄西林（7.5%）、恩诺沙星（7.5%）、庆大霉素（5.0%）、诺氟沙星（2.5%）、环丙沙星（2.5%）、阿莫西林/克拉维酸（1.3%）和头孢噻呋（1.3%）8种抗菌药物耐药,仅对安普霉素敏感。该牛场分离的大肠杆菌对常用抗菌药物耐药情况一般,但中介率较高,须在临床治疗细菌性疾病中避免使用不敏感和中介率高的抗菌药物,养殖场饮水和饲料有被耐药大肠杆菌污染的风险。     

A survey of antimicrobial resistance in Escherichia coli isolated from wild sika deer (Cervus nippon) in Japan

We examined the antimicrobial susceptibility of 848 Escherichia coli isolates from 237 feces samples of wild sika deer (Cervus nippon) captured between 2016 and 2019 in 39 of the 47 prefectures of Japan. Five of the 237 wild sika deer (2.1%) carried E. coli with resistance to at least one antimicrobial, and all the resistant isolates showed resistance to tetracycline. The resistant isolates contained antimicrobial resistance genes that were similar to those in E. coli derived from humans and farm animals. Although wild sika deer are not currently likely to be a source for the transmission of antimicrobial resistance in Japan, they can potentially mediate antimicrobial resistance spread by coming into contact with humans, animals, and their surroundings.     

Antimicrobial resistance of Salmonella and generic Escherichia coli isolated from surface water samples used for recreation and a source of drinking water in southwestern Ontario,Canada

Antimicrobial resistance (AMR) in the aquatic environment represents an important means of introduction and dissemination of resistance genes, and presence of resistant pathogens in surface waters may pose a public health concern to recreational and drinking water users. The purpose of this study was to explore antimicrobial resistance patterns in water samples collected from the Grand River watershed (southwestern Ontario, Canada) to describe the composition, trends and potential risks of AMR in the aquatic environment. As part of FoodNet Canada and the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS), stream water samples were collected bi‐weekly from sampling sites within the Grand River watershed in the Waterloo, Ontario sentinel site and tested for the presence and antimicrobial susceptibility of Salmonella spp. (2005–2013) and generic Escherichia coli (2012–2013). Of all samples tested, 16% of Salmonella and 22% of E. coli isolates were resistant to at least one antimicrobial, including three Salmonella isolates and two E. coli isolates that were resistant to Category I antimicrobials, which are classified as very high importance for the treatment of serious bacterial infections in humans. The greatest proportion of resistant E. coli isolates were observed from the river site upstream of the drinking water intake, while the greatest proportion of resistant Salmonella isolates were from sites upstream in the watershed, and at one recreational water site. Salmonella resistance trends remained fairly stable between 2007 and 2013, with the exception of streptomycin and tetracycline which increased in 2010 and 2013. Continued surveillance of antimicrobial resistance patterns and exploration of risk factor data will allow for a better understanding of resistance transmission in the aquatic environment.     

Pharmacokinetics of ketamine following a short intravenous infusion to isoflurane-anesthetized New Zealand White rabbits (Oryctolagus cuniculus)

ObjectiveTo describe the pharmacokinetics of ketamine following a short intravenous (IV) infusion to isoflurane-anesthetized rabbits.Study designProspective experimental study.AnimalsA total of six adult healthy female New Zealand White rabbits.MethodsAnesthesia was induced with isoflurane in oxygen. Following determination of isoflurane minimum alveolar concentration (MAC), the isoflurane concentration was reduced to 0.75 MAC and ketamine hydrochloride (5 mg kg^–1) was administered IV over 5 minutes. Blood samples were collected before and at 2, 5, 6, 7, 8, 9, 13, 17, 21, 35, 65, 125, 215 and 305 minutes after initiating the ketamine infusion. Samples were processed immediately and the plasma separated and stored at –80 °C until analyzed for ketamine and norketamine concentrations using liquid chromatography–mass spectrometry. Compartment models were fitted to the concentration–time data for ketamine and for ketamine plus norketamine using nonlinear mixed-effects (population) modeling.ResultsA three- and five-compartment model best fitted the plasma concentration–time data for ketamine and for ketamine plus norketamine, respectively. For the ketamine only model, the volume of distribution at steady state (Vss) was 3217 mL kg^–1, metabolic clearance was 88 mL minute^–1 kg^–1 and the terminal half-life was 59 minutes. For the model including both ketamine and norketamine, Vss were 3224 and 2073 mL kg^–1, total metabolic clearance was 107 and 52 mL minute^–1 kg^–1 and terminal half-lives were 52 and 55 minutes for the parent drug and its metabolite, respectively.Conclusions and clinical relevanceThis study characterized the pharmacokinetics of ketamine and norketamine in isoflurane-anesthetized New Zealand White rabbits following short IV infusion. The results obtained herein will be useful to determine ketamine infusion regimens in isoflurane-anesthetized rabbits.     

The in vitro response of field strains of sheep blowflies Lucilia sericata and L. cuprina (Calliphoridae) in New Zealand to dicyclanil and triflumuron

AIM: To evaluate the in vitro efficacy of triflumuron and dicyclanil against field strains of Lucilia sericata and L. cuprina in New Zealand to establish whether anecdotal reports of declining protection periods afforded by dips containing these active ingredients had validity.

METHODS: Strains of field-derived blowflies (Lucilia cuprina and L. sericata) were obtained from adult flies from flytraps or larvae from sheep with flystrike submitted from seven regions of the North and two regions of the South Islands of New Zealand. A total of 27 strains of L. sericata and 19 of L. cuprina were established in culture and first instar larvae obtained were exposed in vitro to dilution series of technical-grade dicyclanil (a pyrimidine carbonitrile) and triflumuron (a benzoyl urea). Concentrations lethal to 50% (LC₅₀) and 95% (LC₉₅) of each strain were calculated and tested by probit analysis, with the ratio of LC₅₀ of each strain divided by LC₅₀ of the most susceptible strain (resistance factor) derived in order to facilitate between-strain comparisons of susceptibility.

RESULTS: There was considerable variation between strains of L. sericata larvae in their susceptibility to triflumuron, with those least susceptible originating from the Waikato region. Strains from both Tararua and Manawatu regions were most susceptible and East Coast strains intermediate. All strains of the species tested showed a narrow band of susceptibility to dicyclanil with no regional trends apparent.

CONCLUSIONS: Representative field-strains of sheep blowflies exhibited a range of susceptibilities to both triflumuron and dicyclanil, suggestive of both genetic heterogeneity among fly populations and the spectrum of dip use in their source areas. This means there should be concerns for the long-term benefits of benzoyl urea insecticides for sheep farmers in New Zealand, but not yet for dicyclanil-based formulations.

CLINICAL RELEVANCE: Flystrike is a debilitating and occasionally lethal disease in sheep such that efficacious sheep dips have prophylactic value in reducing the prevalence of the condition.     

Reduced efficacy of moxidectin and abamectin in young red deer (Cervus elaphus) after 20 years of moxidectin pour-on use on a New Zealand deer farm

A study was undertaken on weaned 4–5 month old farmed red deer to test the efficacy of moxidectin and abamectin anthelmintics, given by three different routes of administration, compared with an untreated control. Faecal samples were collected on days 0, 7 and 14 for a faecal egg count reduction test (FECRT), blood samples were collected on days 0, 0.5, 1, 2, 3, 5, 7, 10 and 14 for pharmacokinetics, and the deer were killed on days 14 or 15 for total nematode count.The control group averaged 1264 adult Ostertagia-type nematode parasite species and treatment efficacy was 77.4% for moxidectin injection, 26% for oral moxidectin and 27.6% for pour-on moxidectin, while the treatment efficacy was 72.4% for abamectin injection, 70.1% for oral abamectin (Hi-Mineral) and 34.1% for pour-on abamectin. Both moxidectin and abamectin injections were significantly more efficacious than their equivalent pour-ons. There was a significant difference in efficacy between oral abamectin (Hi-Mineral) and oral moxidectin (P < 0.01).The control group averaged 2956 adult lungworm (Dictyocaulus eckerti) and 50 Oesophagostomum venulosum in the large intestine and treatment efficacy against these nematodes was 100% for all treatments. There were negligible numbers of other gastro-intestinal nematodes.At slaughter, there was a significant correlation (P = 0.02) between FEC and Ostertagia-type nematodes in the untreated controls. Relatively few eggs were found in faeces from treated animals at 7 and 14 days post-treatment despite significant worm burdens in all six treatment groups, suggesting egg-laying suppression in resistant nematodes, and all three different FECRT calculations tended to overestimate the efficacy of the treatments compared with actual nematode counts.Peak plasma concentrations (C_max) for both actives were measured 12 h after treatment for injection and oral and at 5 days for pour-on. C_max (ng/ml) for moxidectin injection, oral and pour-on were 71.8, 8.3 and 0.4, respectively, and for abamectin injection, oral and pour-on were 62.1, 30.3 and 10.0, respectively. Area under the curve (AUC) estimates for moxidectin injection, oral and pour-on were 106.6, 12.9 and 6.1, respectively, and for abamectin injection, oral and pour-on were 162.7, 57.5 and 74.3, respectively.The results demonstrate that significant anthelmintic resistance to moxidectin and abamectin is present on this deer farm. However, the injection was the most effective route of administration in young deer for both anthelmintics, although <80% efficacious. We conclude that the FECRT is unreliable in deer when anthelmintic resistance is present.     

Diagnosis of tuberculosis due to Mycobacterium bovis in New Zealand red deer (Cervus elaphus) using a composite blood test and antibody assays

A blood test for tuberculosis in deer was developed as an ancillary test to clarify the status of skin test-positive deer, with non-specific sensitisation following exposure to saprophytic mycobacteria. The blood test incorporates the measurement of the relative humoral and cellular immunological responses to Mycobacterium bovis and M. avium antigens to provia composite test with high levels of sensitivity (>95%) and specificity (>98%). The specificity of the test has allowed it to be used in parallel with the skin test to salvage thousands of tuberculosis-free deer with non-specific skin test-positive reactions, while its high sensitivity has consistently identified M. bovis-specific reactivity in tuberculous skin test-positive animals. The rules for establishing the diagnostic parameters for the cellular and antibody assays were developed by retrospective analysis of the laboratory results using blood samples from many thousand tuberculous or disease-free deer. The sensitivity of the blood test was tested in this study using 150 animals with tuberculosis diagnosed by the isolation of M. bovis. It had sensitivity values of 95.7–95.9% in herds with a low (<2.0% ) or a high (>30.0%) incidence of tuberculosis. The test had a specificity of 98.0% when tested on 218 disease-free animals, 118 of which were skin test-positive.

An antibody test was developed to diagnose M. bovis in skin test-negative “anergic” deer from tuberculosis infected herds. When this test was used with deer blood taken 10 days after reading the skin test, it had a sensitivity of 85.3% for 102 M. bovis-positive deer. When used in combination with skin test, the antibody test complemented the skin test to raise the sensitivity of the combined tests to 95.0%) when antibody-positive or skin test-positive tests were used to diagnose tuberculosis. The specificity of the antibody test was 100% when used to evaluate 218 disease-free deer from non-infected herds.     

Phenotypic and genotypic characteristics of antimicrobial resistant Escherichia coli isolated from symbovine flies, cattle and sympatric insectivorous house martins from a farm in the Czech Republic (2006-2007)

The prevalence of antimicrobial resistant Escherichia coli was tested in symbovine flies and sympatric house martins (Delichon urbica) at a dairy farm. Antimicrobial resistant E. coli was detected in 89% (n = 147) of isolates from flies within a calf barn. Isolates with the same antimicrobial resistance phenotypes, genes, and pulsotypes were found between both fly and calf E. coli isolates, suggesting that the calves were the initial source of the antimicrobial resistant strains in fly isolates. Symbovine flies were considered as important reservoirs of antimicrobial resistant E. coli strains at a dairy farm, due to their intensive contact with cattle feces and manure. House martin fecal samples from the same farm contained 4.5% (n = 393) of antimicrobial resistant E. coli. House martin isolates displayed different macrorestriction profiles than fly isolates and the significance of house martins as a reservoir and vector of antimicrobial resistant E. coli appears low.