Etiology and antimicrobial susceptibility of udder pathogens from cases of subclinical mastitis in dairy cows in Sweden

Background

A nationwide survey on the microbial etiology of cases of subclinical mastitis in dairy cows was carried out on dairy farms in Sweden. The aim was to investigate the microbial panorama and the occurrence of antimicrobial resistance. Moreover, differences between newly infected cows and chronically infected cows were investigated.

Methods

In total, 583 quarter milk samples were collected from 583 dairy cows at 226 dairy farms from February 2008 to February 2009. The quarter milk samples were bacteriological investigated and scored using the California Mastitis Test. Staphylococci were tested for betalactamase production and presence of resistance was evaluated in all specific udder pathogens. Differences between newly infected cows and chronically infected cows were statistically investigated using logistic regression analysis.

Results

The most common isolates of 590 bacteriological diagnoses were Staphylococcus (S) aureus (19%) and coagulase-negative staphylococci (CNS; 16%) followed by Streptococcus (Str) dysgalactiae (9%), Str. uberis (8%), Escherichia (E.) coli (2.9%), and Streptococcus spp. (1.9%). Samples with no growth or contamination constituted 22% and 18% of the diagnoses, respectively. The distribution of the most commonly isolated bacteria considering only bacteriological positive samples were: S. aureus - 31%, CNS - 27%, Str. dysgalactiae - 15%, Str. uberis - 14%, E. coli - 4.8%, and Streptococcus spp. - 3.1%. There was an increased risk of finding S. aureus, Str. uberis or Str. dysgalactiae in milk samples from chronically infected cows compared to findings in milk samples from newly infected cows. Four percent of the S. aureus isolates and 35% of the CNS isolates were resistant to penicillin G. Overall, resistance to other antimicrobials than penicillin G was uncommon.

Conclusions

Staphylococcus aureus and CNS were the most frequently isolated pathogens and resistance to antimicrobials was rare.     

Antimicrobial susceptibility of mastitis pathogens from first lactation and older cows

Increasing antimicrobial resistance has become a serious concern worldwide and antimicrobial use in animal agriculture is currently under scrutiny. Mastitis is the most common reason for antibiotic use in dairy herds and thus, antimicrobial resistance of mastitis pathogens has received recent attention. The purpose of this study was to describe and compare antimicrobial susceptibility of mastitis pathogens isolated at calving from first lactation and older cows. A total of 202 bacteria were isolated from intramammary infections (IMIs) within 3 days after calving over a 16-month study period in the Krauss Dairy Research Herd at the Ohio Agricultural Research and Development Center (OARDC) in Wooster, OH. Of these IMIs, 78% were caused by coagulase-negative staphylococci (CNS). Forty-four percent of them were resistant to at least one antibiotic. Most resistance was observed against penicillin, 39% of the isolates from older cows and 26% from first lactation cows being resistant to penicillin (P > 0.05). Also MIC90 for penicillin was higher among isolates from older cows. On the other hand, resistance to tetracycline was more common and MIC90 higher among isolates from first lactation cows than from older cows. Differences in the proportions of resistant isolates between first lactation and older cows were not statistically significant, though. The resistance patterns of the CNS isolated during the study are concordant with antimicrobial usage in the study herd. This is in agreement with the generally accepted notion that selection pressure from the use of antibiotics is a main factor in development of antibiotic resistance.     

Use of antimicrobial susceptibility testing of bacterial pathogens isolated from the milk of dairy cows with clinical mastitis to predict response to treatment with cephapirin and oxytetracycline

OBJECTIVE: To determine whether results of antimicrobial susceptibility testing of bacterial pathogens isolated from the milk of dairy cows with clinical mastitis were associated with duration of clinical signs or bacteriologic cure rate following treatment with cephapirin and oxytetracycline. DESIGN: Observational study on a convenience sample. ANIMALS: 58 dairy cows with 121 episodes of clinical mastitis. PROCEDURE: Cows that only had abnormal glandular secretions were treated with cephapirin alone. Cows with an inflamed gland and abnormal glandular secretions were treated with oxytetracycline and cephapirin. Cows with systemic signs of illness, an inflamed gland, and abnormal glandular secretions were treated with oxytetracycline and flunixin meglumine and frequent stripping of the affected glands. The Kirby-Bauer method was used for antimicrobial susceptibility testing, and current guidelines were used to categorize causative bacteria as susceptible or resistant to the treatment regimen. RESULTS: Median durations of episodes of clinical mastitis caused by susceptible (n = 97) and resistant (24) bacteria were not significantly different. Bacteriologic cure rates at 14 and 28 days were similar for episodes caused by susceptible and resistant bacteria; however, for 56 episodes of clinical mastitis caused by gram-positive bacteria and treated with cephapirin alone, bacteriologic cure rate at 28 days was significantly higher for susceptible than for resistant bacteria. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that antimicrobial susceptibility testing was of no value in predicting duration of clinical signs or bacteriologic cure rate in dairy cows with mastitis, except for episodes caused by gram-positive organisms treated with intramammary administration of cephapirin alone.     

Treatment of clinical mastitis. Using antimicrobial susceptibility profiles for treatment decisions.

Antibiotic susceptibility of clinical mastitis pathogens has traditionally been determined using the agar diffusion method that was designed to reflect the antibiotic concentration in serum and interstitial fluid of human patients after receiving oral or intravenous administration. The validity of applying agar diffusion susceptibility breakpoints derived from humans to the treatment of bovine mastitis has not been established and is extremely questionable because (1) bovine milk pH and electrolyte, fat, protein, and leukocyte concentrations, growth factor composition, and pharmacokinetic profiles are different than those for human plasma and (2) human bacterial pathogens are often different from bovine mastitis pathogens. Also, antibiotics are distributed unevenly in an inflamed gland, and high antibiotic concentrations can alter neutrophil morphology or function in vitro and thereby inhibit bacterial clearance in vivo. The current cost of antibiotic susceptibility testing is $12 to $20 per test. Because the dairy industry is economically driven, any diagnostic test should be validated, have appropriate sensitivity and specificity, and have an acceptable economic return on the cost of testing before it can be routinely recommended. In the authors' opinion, antimicrobial susceptibility testing of mastitis pathogens has not been adequately validated for most mastitis pathogens and antibiotics; therefore, the authors do not currently recommend the use of susceptibility testing to guide treatment decisions for individual cows. Additional research is needed to further define the role, if any, that antimicrobial susceptibility testing should play in the treatment of clinical mastitis.     

Relationship between somatic cell count status and subsequent clinical mastitis in Dutch dairy cows

High composite somatic cell counts (CSCC) in dairy cows may develop into clinical mastitis (CM), suggesting that prevention or intervention of high CSCC may prevent CM later in lactation. The objective of this study was to quantify the relationship between high CSCC in dairy cows and the first subsequent case of CM in the same lactation. Farmer-diagnosed cases of CM and test day CSCC measurements during 1 year of 13,917 cows in 196 randomly selected Dutch dairy herds were available for analysis. Cows were followed in 1 lactation from the first test day postpartum until CM, drying off, culling or end of study. Cox proportional hazards models with time-varying CSCC levels were used to estimate the effect of high CSCC (≥200,000cells/ml) on the time until the first case of CM. A shared frailty effect was included to adjust for clustering of cows within herds. The proportion of cows developing CM after a CSCC measurement was 11%. Primiparae with a high CSCC had a 4-fold higher hazard for subsequent CM than primiparae with a low CSCC; multiparae with a high CSCC had a 2-fold higher hazard than multiparae with a low CSCC. Additionally, multiparae with a low CSCC had a 2-fold higher hazard for CM occurrence than primiparae with a low CSCC. Increasing the threshold for high CSCC showed that the risk for CM increased. If the last CSCC before CM was low, CSCC information of 2 preceding test days was more predictive than CSCC information from only the last test day. When the last CSCC was high, CSCC information of 2 preceding test days did not have added predictive value. This study identified that approximately 25% of first subsequent CM cases after a CSCC measurement can potentially be prevented when cows are prevented to get high CSCC or when high CSCC cows are removed from the population. This corresponded with a decrease in the proportion of lactating cows with CM after a CSCC measurement from 11% to 7%.     

Relationship between teat morphological traits and subclinical mastitis in Frieswal dairy cows

The present study was aimed to investigate the relationship between selected morphological traits of teat and subclinical mastitis (SCM) in Frieswal crossbred dairy cows. A total of 1040 quarters from 261 lactating cows were evaluated for teat shape (bottle/fleshy/collapsed/conical/normal/pencil and short), teat-end shape (dished/flat/funnel/pocketed/pointed and rounded), teat orientation (aligned/misaligned) and teat position (front and rear; left-sided and right-sided). Each udder quarter was screened with California Mastitis Test (CMT) for the purpose of defining quarter health status. Data were analysed using Chi-square test and multivariable logistic regression procedure. An overall prevalence of SCM (CMT positive) at quarter level was 30.6%. Most of the teats had normal or cylindrical shapes (48%), dished teat-ends (40.7%), and aligned (central or squared) in orientation (65%). At bivariable level, significant association of SCM with teat shape, teat position, teat orientation, parity, and stage of lactation was observed (P < 0.05 to P < 0.001). Teat-end shapes showed some association with SCM (P = 0.07). Results of multivariable analysis showed that pencil-shaped teats were least associated with SCM (P < 0.05) as compared to other teat shapes. Prevalence of SCM was also higher in rear teats (P = 0.015), misaligned teats (P = 0.01), and cows in second or higher parities (P < 0.01) and late stage of lactation (P < 0.001). The results of the present study indicate that selected morphological traits of teat are associated with SCM in Frieswal crossbred cows; therefore, selection towards desirable morphological traits could help reduce mastitis in this breed.     

Rates of resistance of mastitis pathogens from cows in Switzerland

1215 strains of bacteria isolated from cows suffering from acute or chronic clinic mastitis were tested in the antibiogram according to the Kirby-Bauer method. The germ spectrum included: 304 DNase-positive strains of Staphylococci 304 DNase-negative strains of Staphylococci 304 CAMP-negative strains of Streptococci 303 Strains of Enterobacteriaceae The antibiotics selected were Penicillin, Cloxacillin, Neomycin und Gentamicin, then the combinations Penicillin/Neomycin, Cloxacillin/Gentamicin and Nafcillin/Penicillin/Dihydrostreptomycin. Gentamicin showed excellent action against Staphylococci and Enterobacteriaceae. Further on, the present investigations show, that the combination Cloxacillin/Gentamicin is fully effective in the gram-positive spectrum (against Streptococci and Staphylococci), as in the gram-negative spectrum (against E. coli).     

生物氧疗法辅助治疗奶牛临床型乳房炎疗效观察

乳房炎是成年奶牛最常见的一种疾病,发病率可达30%(Gardner等,1990),奶牛生产单位每年因乳房炎导致的淘汰率约10%,占牛群总淘汰率的1/2左右[1].乳房炎可以降低奶牛产奶量(下降10%～15%)、增加医疗费用支出,从而造成严重的经济损失;此外,乳房炎还会降低牛奶品质、造成牛奶的公共卫生安全隐患[1-3].     

In vitro susceptibility of Staphylococcus aureus strains isolated from cows with subclinical mastitis to different antimicrobial agents

Sensitivity to commercial teat dips (nonoxinol-9 iodine complex and chlorhexidine digluconate) of 56 Staphylococcus (S.) aureus strains isolated from quarter milk samples of various German dairy herds treated with different teat dipping schemes was investigated in this study. The minimum inhibitory concentration was determined using a broth macrodilution method according to the German Veterinary Association guidelines. The main objective of the current study was to induce in vitro resistance induction of S. aureus to chemical disinfectants. Ten different strains were repeatedly passed ten times in growth media with sub-lethal concentrations of disinfectants. Nine strains showed a significant reduction in susceptibility to the nonoxinol-9 iodine complex but only one strain developed resistance to chlorhexidine digluconate. Stability of the acquired resistance was observed in all S. aureus strains adapted to the nonoxinol-9 iodine complex and chlorhexidine digluconate. In contrast, simultaneous resistance to different antibiotics was not observed in any of the ten investigated S. aureus strains. However, the isolates exhibited a high degree of resistance to penicillin G. Based on these results, resistance of S. aureus to chemical disinfectants may be more likely to develop if the chemicals are used at concentrations lower than that required for an optimal biocidal effect.     

Differences in mastitis susceptibility between Holstein Friesian,Dutch Friesian and Dutch Red and White cows

Summary

On an experimental farm 3 dairy breeds were compared with regard to mastitis by various parameters.

Higher cell counts and more udder infections were present in the Dutch Red and White (DRW) in comparison with the Holstein Friesian (HF) and the Dutch Friesian (DF). Within the HF breed a negative correlation between cell count and production was established and also in this breed, a negative correlation between maximum rate of milk flow and cell count.

There is no evidence that higher procuction and easier milking are connected with more mastitis. On the contrary, within the breeds an opposite trend appeared. Selection against mastitis does not conflict with selection for production and ease of milking.

An addition trial revealed that sampling after too quick preparation of the cows before milking resulted in much higher cell counts in quarter samples.

Cows which were selected for drying off with antibiotics recovered from their infection but these more susceptible animals maintained a higher cell count to a certain extent.     

Differences in mastitis susceptibility between Holstein Friesian, Dutch Friesian and Dutch Red and White cows

Summary On an experimental farm 3 dairy breeds were compared with regard to mastitis by various parameters. Higher cell counts and more udder infections were present in the Dutch Red and White (DRW) in comparison with the Holstein Friesian (HF) and the Dutch Friesian (DF). Within the HF breed a negative correlation between cell count and production was established and also in this breed, a negative correlation between maximum rate of milk flow and cell count. There is no evidence that higher procuction and easier milking are connected with more mastitis. On the contrary, within the breeds an opposite trend appeared. Selection against mastitis does not conflict with selection for production and ease of milking. An addition trial revealed that sampling after too quick preparation of the cows before milking resulted in much higher cell counts in quarter samples. Cows which were selected for drying off with antibiotics recovered from their infection but these more susceptible animals maintained a higher cell count to a certain extent.     

In vitro activity of 10 antimicrobial agents against bacteria isolated from cows with clinical mastitis

The susceptibility of 495 strains of bacteria, recently isolated in France from cows with clinical mastitis, to 10 antimicrobial agents--penicillin G, cloxacillin, oxacillin, cephalexin, cefazolin, cephapirin, cefquinome, neomycin, ampicillin and colistin--was determined by measuring their minimum inhibitory concentrations (MICS). Overall, the levels of resistance were very low except for staphylococci and penicillin G. The 167 streptococcal strains were susceptible to all of the beta-lactams tested, but six (3-6 per cent) were highly resistant to neomycin. Of the 171 staphylococcal isolates, 36.2 per cent were resistant to penicillin G, one strain of Staphylococcus sciuri was classified as methicillin-resistant, but they were all susceptible to neomycin. None of the 122 strains of Escherichia coli was resistant to colistin, but 12 had high MIC values for one or more of the cephalosporins.     

Evaluation of stratification factors and score-scales in clinical trials of treatment of clinical mastitis in dairy cows

There is often a need to reduce sample size in clinical trials due to practical limitations and ethical considerations. Better comparability between treatment groups by use of stratification in the design, and use of continuous outcome variables in the evaluation of treatment results, are two methods that can be used in order to achieve this. In this paper the choice of stratification factors in trials of clinical mastitis in dairy cows is investigated, and two score-scales for evaluation of clinical mastitis are introduced. The outcome in 57 dairy cows suffering from clinical mastitis and included in a clinical trial comparing homeopathic treatment, placebo and a standard antibiotic treatment is investigated. The strata of various stratification factors are compared across treatments to determine which other factors influence outcome. The two score scales, measuring acute and chronic mastitis symptoms, respectively, are evaluated on their ability to differentiate between patients classified from clinical criteria as responders or non-responders to treatment. Differences were found between the strata of the factors severity of mastitis, lactation number, previous mastitis this lactation and bacteriological findings. These factors influence outcome of treatment and appear relevant as stratification factors in mastitis trials. Both score scales differentiated between responders and non-responders to treatment and were found useful for evaluation of mastitis and mastitis treatment.     

Effects of oral selenium supplementation on mastitis markers and pathogens in Estonian cows

The effects of selenium supplementation on mastitis parameters in milk and on glutathione peroxidase (GPx) levels in blood were evaluated. Fifty-five Estonian dairy cows were allocated to selenium-supplemented (n=39) and nonsupplemented (n=16) groups. The supplemented group received 0.2 ppm organic selenium in the form of selenium yeast in their diet daily for 8 weeks. The nonsupplemented cows received their standard diet with no selenium supplementation. Mastitis parameters (i.e., bacteriologic findings and somatic cell count, N-acetyl-beta-D-glucosaminidase, and bovine serum albumin concentration) and GPx levels were monitored. The increase in the activity of GPx was significantly (P<.001) greater in selenium-supplemented cows than in nonsupplemented ones. Milk samples from each quarter were examined before and 8 weeks after initiation of the study. The proportion of quarters still pathogen-free after 8 weeks was significantly (P<.01; odds ratio [OR]=9.7) higher in selenium-supplemented cows than in nonsupplemented cows. However, when quarters milk-tested as pathogen-infected at the start of the study were reexamined after 8 weeks, there was no significant (P=.14; OR 3.6) difference in the proportion of pathogen-free quarters between selenium-supplemented cows and nonsupplemented cows. Differential positive rate (Youden's index) revealed that individual quarters were more prone to be infected by pathogens when the blood GPx activities in cows were below the cutoff value of 3.3 microkat/g hemoglobin than when GPx activity was above this value. It was concluded that selenium supplementation in cows with low GPx activity seems to support udder defense mechanisms that favor reduction of the incidence of new mastitis cases.     

Prevalence, bacterial causes, and antimicrobial susceptibility profile of mastitis isolates from cows in large-scale dairy farms of Northern Ethiopia

The study was undertaken to determine the prevalence of bovine mastitis, isolate mastitis causing bacteria, assess the association of some risk factors, and determine the antibiotic resistance pattern of bacterial isolates in cows in large-scale dairy farms of Northern Ethiopia. A total of 305 lactating and nonlactating cows were included in the present study. The overall prevalence of clinical and subclinical mastitis was 3.6 and 33.8?%, respectively. The quarter level prevalence was 15.4?%; from which, 11.9 and 1.1?% were subclinical form and blind teat, respectively, while the remaining 2.4?% were of clinical form. Staphylococcus aureus accounted for 36?% of the isolates followed by Escherichia coli (27.3?%). Risk factors including age (p?9?years (OR?=?3.63, 95?% CI?=?1.42-9.25)), cows with four to seven calves (OR?=?3.39, 95?% CI?=?2.06-5.60), and cows in late lactation stage (OR?=?3.79, 95?% CI?=?1.64-8.75). In multivariable logistic regression analysis, age (p?=?0.005) and lactation stage (p?=?0.027) showed statistically significant association with the occurrence of mastitis. The antimicrobial susceptibility pattern showed high susceptibility of S. aureus to nalidixic acid (82.4?%) followed by chloramphenicol (58.8?%); however, these species were resistant to the rest of the antimicrobials tested. Highest resistance was observed against clindamycin and ampicillin. Coliform bacteria (E. coli and Klebsiella pneumoniae) showed resistance to most of the antimicrobials used. Detailed investigation is needed to identify the interplay of managemental and environmental risk factors to design appropriate control measures.     

Prevalence of subclinical mastitis and isolated udder pathogens in dairy cows in Southern Vietnam

Dairy production is not traditional in Vietnam. The farmers have little practical knowledge and udder health control is generally lacking. In order to give the farmers appropriate advice, knowledge about the distribution of udder pathogens is crucial. The aim of the study was to investigate the prevalence of sub-clinical mastitis and to identify udder pathogens isolated from smallholder dairy herds in Southern Vietnam. Twenty farms with a herd somatic cell count (SCC) ranging from low (≤400?×?10³?cells/mL) to high (>400?×?10³?cells/mL) were randomly selected. Milk samples were collected from 458 quarters of 115 clinically healthy cows. SCC was analyzed on farm by a portable cell counter. Bacteriological samples were taken using Mastistrip^© cassettes and sent to Sweden for examination. For all herds the mean herd SCC was 632?×?10³/mL milk. The prevalence of subclinical mastitis at quarter SCC basis was 63.2 % and at cow basis 88.6 %. Only 40 % of all cows were bacteriologically negative in all quarters. Streptococcus agalactiae was the most commonly found bacteria species, isolated from 96 of the 458 quarter samples, in 13 of the 20 farms. The results indicate pronounced subclinical mastitis problems among the dairy cows in this region mainly due to infections with S. agalactiae. The high prevalence of this highly contagious pathogen is probably attributable to the generally poor milking hygiene and low awareness of proper measures to prevent occurrence and spread of udder infections. A strict, targeted action program for the herds in this area is required in order to lower the prevalence of subclinical mastitis.