A note on effectiveness of dry cow therapy in New Zealand dairy herds

Two dry-cow therapy products were evaluated in seven factory-supply dairy herds in the Waikato area. A product containing neomycin sulphate and the benzathine salt of penicillin (Neopen D.C. White; Smith-Biolab) was used in five herds, and one containing benzathine cloxacillin (Orbenin, Beecham) was used in two herds. Non-treated control cows were included in each herd. Both products were effective in eliminating intramammary infections with Staphylococcus aureus, Streptococcus uberis, and Streptococcus agalactiae. Efficacy of dry-cow therapy against S. aureus was 83.8% and 85.2% respectively. Spontaneous cure rate among controls was 30.8% for S. aureus during the dry period. Spontaneous cure rate for Str. uberis was 50%, while dry-cow therapy eliminated 100% and 77.8%, respectively, for the two products. Dry-cow therapy with either product eliminated more than 90% of Str. agalactiae infections while spontaneous cure rate was only 28.6%. These results further support the effectiveness of dry-cow therapy in reducing the level of subclinical mastitis in dairy herds by shortening the duration of intramammary infections.     

Effects of glycolytic and cytoskeletal inhibitors on phagocytic and nitroblue tetrazolium reductive activities of bovine neutrophils

Phagocytic and oxidative metabolic activities of bovine blood neutrophils were determined in the presence of glycolytic (NaF) and cytoskeletal (colchicine, cytochalasin B, and prostaglandin E1) inhibitors. Phagocytosis and post-phagocytic oxidative metabolic activity, measured by nitroblue tetrazolium reduction, were determined using zymosan, Escherichia coli, Staphylococcus aureus, or Streptococcus agalactiae. Sodium fluoride (1.25 microM to 1.25 mM concentrations) did not significantly (P greater than 0.05) inhibit phagocytosis of S aureus and Str agalactiae, whereas phagocytosis of zymosan and E coli was significantly (P less than 0.05) inhibited only at 1.25 mM concentration. Colchicine at 1.25 nM to 1.25 microM concentrations significantly inhibited phagocytosis of zymosan and E coli, but not of S aureus and Str agalactiae. Cytochalasin B at 1.25 nM to 1.25 microM concentrations significantly inhibited phagocytosis of zymosan and all 3 bacteria, whereas prostaglandin E1 was noninhibitory at similar concentrations. Nitroblue tetrazolium reduction, in general, was not significantly affected by NaF and cytoskeletal inhibitors.     

Herd management and prevalence of mastitis in dairy herds with high and low somatic cell counts

Thirty-two dairy herds, 16 with low somatic cell counts (LSCC; Dairy Herd Improvement Association 12-month mean herd SCC less than or equal to 150,000 cells/ml) and 16 with high somatic cell counts (HSCC; Dairy Herd Improvement Association 12-month mean herd SCC greater than or equal to 700,000 cells/ml) were evaluated to determine the relationship between the prevalence of mastitis in each herd and each herd's mastitis control and management practices. Once for each herd, duplicate quarter milk samples were collected from the lactating cows, a survey of herd mastitis control, milking hygiene, and management practices of each herd was performed, and milking-machine function was evaluated. Of the 16 herds with LSCC, 2 (12.5%) had Streptococcus agalactiae isolated and 7 (44%) had Staphylococcus aureus isolated. Both organisms were found in all of the herds with HSCC. In herds with LSCC, the mean percentage of quarters infected with Str agalactiae was 0.1%, the mean percentage infected with streptococci other than Str agalactiae was 1.9%, and the mean infected with S aureus was 0.7%. In herds with HSCC, 25.7% of the quarters were infected with Str agalactiae, 3.7% were infected with streptococci other than Str agalactiae, and 7.6% were infected with S aureus. A program of postmilking teat dipping and treatment of all cows at the beginning of the nonlactating period was practiced more frequently in the herds with LSCC (81.3%) than in the herds with HSCC (37.5%). Major differences were not found between the 2 groups of herds in the use of the more common milking hygiene techniques or in the maintenance and functional characteristics of the milking equipment.     

Mannitol agar for microbiologic diagnosis of bovine mastitis

A medium containing mannitol (mannitol agar) was developed and evaluated as a tool for the microbiologic diagnosis of bovine mastitis. Mannitol agar supported growth of all important bacterial mastitis pathogens (staphylococci, streptococci, coliforms, and pseudomonads) except Corynebacterium pyogenes. Color change around colonies in the agar permitted the differentiation of pathogenic from nonpathogenic staphylococci. Most Staphylococcus aureus strains and some Staphylococcus epidermidis strains produced yellow zones. These yellow zone-producing strains (mannitol fermenters) of staphylococci were obtained from quarters with significantly elevated (P less than 0.05) somatic cell counts (SCC) in the milk, as compared with uninfected quarters and, therefore, would be considered pathogens. Mannitol-negative strains of S epidermidis (those with red zones) were obtained from quarters with SCC similar to those of uninfected quarters. The streptococci could be divided into 2 groups on the basis of color change around the colonies: Streptococcus agalactiae, Str dysgalactiae, and group G streptococci produced red zones; Str uberis, Str bovis, and enterococci produced yellow zones. Pathogenic streptococci (Str agalactiae, Str dysgalactiae, Str uberis, and group G streptococci) were obtained from quarters with SCC significantly higher (P less than 0.01) than those of uninfected quarters. Streptococcus bovis and enterococci were obtained from quarters with SCC similar to those of uninfected quarters and were considered nonpathogenic. Pathogenic streptococci were found in much higher concentration than nonpathogenic streptococci and could be differentiated on that basis.     

THE NEW SOUTH WALES MASTITIS CONTROL PROGRAM

Bacteriological examinations were made on quarter samples from cows in 35 herds over a 3 year period to monitor the response in a mastitis control program. Initially, Staphylococcus aureus predominated in 32 of the herds and the mean herd prevalence was 26%. The control measures halved this rate but there was considerable variation in response between herds. The decline occurred rapidly and there was a significant reduction (P less than 0.01) by 3 months. Streptococcus agalactiae predominated in 3 herds and the overall infection rate was 4.9%. Control measures eliminated the infection completely from most herds but reinfection occurred in 2 herds. The greatest decline occurred in the first 6 months and was significant (P less than 0.05). The measures had little effect upon Str. uberis and Str. dysgalactiae which remained fairly consistently at low levels. Initially, strains of Staph. aureus resistant to penicillin were dominant in most herds. In a minority of herds strains resistant to streptomycin predominated and in these herds there was a concurrent resistance to penicillin. These patterns did not change greatly over the control period. Resistance by Str. agalactiae to streptomycin occurred in most herds at the start of the program.     

Bovine mastitis and its association with selected risk factors in smallholder dairy farms in and around Bahir Dar,Ethiopia

Three hundred fifty one (195 local zebu and 156 Holstein x local zebu crosses) lactating cows of smallholder farms in Bahir Dar 'milk shed' were examined from September 2003 to March 2004 to determine mastitis prevalence, isolate pathogens and identify the role of some potential risk factors. Clinical prevalence was determined through examination of abnormalities of milk, udder or cow. California mastitis test (CMT) was used for determination of subclinical mastitis prevalence. Clinical prevalence at cow level was 3.9% in crossbreds and none in local zebu breeds. Subclinical mastitis at cow level based on CMT was high (34.4%) in crossbreds compared to indigenous zebu (17.9%) (p < 0.05). Quarter subclinical prevalence based on CMT was 17.9% and 4.9% in crossbreds and local zebu, respectively. The pathogens isolated from mastitic milk (CMT positive milk) were coagulase negative staphylococci (CNS), S. aureus, Str. agalactiae, Str. dysgalactiae, Str. uberis, Micrococcus species, C. bovis, A. pyogens, B. cereus, and S. intermedius. Among these, the most frequent isolates were CNS (50%), S. aureus (19%), Str. agalactiae (8%) and Str. dysgalactiae (7%). Among potential risk factors considered, stage of lactation, parity and breed were found to affect the occurrence of mastitis significantly (p < 0.05).     

Incidence and types of clinical mastitis in dairy herds with high and low somatic cell counts

Eighteen dairy herds were studied, 12 with a 12-month Dairy Herd Improvement Association herd mean somatic cell count (SCC) less than or equal to 150,000 cells/ml (low SCC) and 6 with a 12-month mean SCC greater than 700,000 cells/ml (high SCC). At the outset of the study, quarter samples for bacteriologic culture were collected (in duplicate) from all quarters of all lactating cows (whole herd culture). Subsequently, quarter milk samples for culture from all cows with clinical mastitis were collected for a period of 6 months. In the herds with low SCC, results of whole herd culture revealed low prevalence of intramammary infection attributable to all major pathogens (less than 4% of all quarters). Prevalence of infection with Streptococcus agalactiae (22.2% of all quarters) and Staphylococcus aureus (6.6% of all quarters) was significantly (P less than 0.05) higher in the herds with high SCC. Mean incidence of clinical mastitis in the herds with low SCC was 4.23 infections/100 cows/month (range, 0.42 to 10.25 infections). In the herds with high SCC, mean incidence was 2.91 infections/100 cows/month (range, 1.33 to 3.92 infections). In the herds with low SCC, infection type, as mean percentage of total clinically infected quarters sampled for culture/herd, was 0.0%, 2.2%, 12.3%, 43.5%, and 28.6% for Str agalactiae, S aureus, streptococci other than Str agalactiae, coliforms, and organisms not isolated, respectively. Respective percentages for the herds with high SCC were 41.5%, 18.3%, 12.6%, 8.0%, and 8.8%. During the study period (from April through January), incidence of clinical mastitis and clinical mastitis caused by coliform bacteria were highest in July and August for herds with low SCC.(ABSTRACT TRUNCATED AT 250 WORDS)     

THE PREVALENCE OF UDDER INFECTION AND MASTITIS IN HERDS PRODUCING BULK MILK WITH EITHER CONSISTENTLY HIGH OR LOW CELL COUNT

Quarter samples from twenty-five dairy herds, representing 223 herds supplying direct to Brisbane, were cultured and submitted to the Wisconsin Mastitis Test (WMT). Thirteen herds had a history of producing bulk milk with a consistently high WMT score (greater than 15 mm) and in twelve herds the WMT score was consistently low. Prevalence of infection was higher in group A herds (22.2% quarters were infected with Staphylococcus aureus or streptococci) than in group B herds (9.8% of quarters were infected). There was considerable scatter of prevalence among both groups of herds. Overall, S. aureus was found in 19% of cows and 7% of quarters, and Streptococcus agalactiae in 15% of cows and 7% of quarters. Quarters from group A herds showed a higher WMT score than those in group B herds whether infected with S. aureus, Str. agalactiae, micrococci or yielded no detectable organisms. It was concluded that regular surveys are required of the high cell count herds to monitor mastitis status in the industry.     

Sensitivity and specificity of latex agglutination tests used to identify Streptococcus agalactiae and Staphylococcus aureus isolated from bulk tank milk

Comparisons were made among rapid latex agglutination tests and conventional biochemical tests used to identify Streptococcus agalactiae and Staphylococcus aureus. Ninety-eight streptococci and 149 staphylococci isolated from bulk tank milk were tested. Sensitivity and specificity for the latex agglutination test used for identification of Str agalactiae were 97.6 and 98.2%, respectively. Sensitivity and specificity for the latex agglutination test used for identification of S aureus were 90.2 and 67.5%, respectively. Of 25 staphylococci considered false-positive by the latex agglutination test, 14 (56%) were considered tube coagulase-positive. Fifteen staphylococci considered false-positive by latex agglutination test had biotypes representative of S hyicus of S xylosus.     

Udder pathogens and their resistance to antimicrobial agents in dairy cows in Estonia

Background

The goal of this study was to estimate the distribution of udder pathogens and their antibiotic resistance in Estonia during the years 2007-2009.

Methods

The bacteriological findings reported in this study originate from quarter milk samples collected from cows on Estonian dairy farms that had clinical or subclinical mastitis. The samples were submitted by local veterinarians to the Estonian Veterinary and Food Laboratory during 2007-2009. Milk samples were examined by conventional bacteriology. In vitro antimicrobial susceptibility testing was performed with the disc diffusion test. Logistic regression with a random herd effect to control for clustering was used for statistical analysis.

Results

During the study period, 3058 clinical mastitis samples from 190 farms and 5146 subclinical mastitis samples from 274 farms were investigated. Positive results were found in 57% of the samples (4680 out of 8204), and the proportion did not differ according to year (p > 0.05). The proportion of bacteriologically negative samples was 22.3% and that of mixed growth was 20.6%. Streptococcus uberis (Str. uberis) was the bacterium isolated most frequently (18.4%) from cases of clinical mastitis, followed by Escherichia coli (E. coli) (15.9%) and Streptococcus agalactiae (Str. agalactiae) (11.9%). The bacteria that caused subclinical mastitis were mainly Staphylococcus aureus (S. aureus) (20%) and coagulase-negative staphylococci (CNS) (15.4%). The probability of isolating S. aureus from milk samples was significantly higher on farms that had fewer than 30 cows, when compared with farms that had more than 100 cows (p < 0.005). A significantly higher risk of Str. agalactiae infection was found on farms with more than 600 cows (p = 0.034) compared with smaller farms. The proportion of S. aureus and CNS isolates that were resistant to penicillin was 61.4% and 38.5%, respectively. Among the E. coli isolates, ampicillin, streptomycin and tetracycline resistance were observed in 24.3%, 15.6% and 13.5%, respectively.

Conclusions

This study showed that the main pathogens associated with clinical mastitis were Str. uberis and E. coli. Subclinical mastitis was caused mainly by S. aureus and CNS. The number of S. aureus and Str. agalactiae isolates depended on herd size. Antimicrobial resistance was highly prevalent, especially penicillin resistance in S. aureus and CNS.     

Recovery of Streptococcus agalactiae and Staphylococcus aureus from fresh and frozen bovine milk

Three hundred seventy-three milk samples were screened for Streptococcus agalactiae and Staphylococcus aureus. After sample storage at -20 C for 23 days, the frequency of Str agalactiae isolation increased 2.50 times. The frequency of S aureus isolation increased 1.48 times in the same interval. Increases in the proportion of these isolates were highly significant (P = 0.000006 and 0.0001, respectively). Results of the study indicate that optimal procedures for microbiological testing for these mastitis pathogens may include preculture freezing. The magnitude of the increase in the proportion of isolates indicates the existence of an important population of infected cattle shedding bacteria at concentrations not detected by use of standard microbiological techniques.     

Effects of source and washing of erythrocytes on growth of bacterial pathogens from the bovine mammary gland

Effects of source and washing of RBC on quantitative growth and hemolytic zone sizes of common bacterial pathogens of the bovine mammary gland were evaluated. Blood samples used to prepare the blood agar media were obtained from 10 adult dairy cows, 10 dairy calves, and 10 sheep. Hemolytic zone sizes produced by Staphylococcus aureus were significantly (P less than 0.01) larger on blood agar prepared with washed RBC than on blood agar prepared with nonwashed RBC, regardless of RBC source. With the exception of Corynebacterium bovis, growth of all bacteria was equivalent or significantly higher on medium prepared with washed RBC, compared with that on medium prepared with nonwashed RBC, regardless of RBC source. Significantly higher numbers of C bovis (P less than 0.01) and Streptococcus agalactiae (P less than 0.01) were isolated on medium prepared with washed cow RBC. Significantly higher numbers of Str uberis (P less than 0.01) and S aureus (P less than 0.05) were isolated on medium prepared with washed sheep RBC and washed calf RBC, respectively. Growth of Escherichia coli was not affected by the RBC source. Seemingly, RBC used in the preparation of medium should be washed. The source of RBC, as well as inter-animal variation, also should be considered in the quality control of medium.     

Intramammary infections in a dairy herd with a low incidence of Streptococcus agalactiae and Staphylococcus aureus infections.

In a dairy herd with a low incidence of intrammary infections due to Streptococcus agalactiae and Staphylococcus aureus, clinical mastitis remained a serious problem despite good control of nonclinical mastitis through postmilking teat disinfection and antibiotic therapy of known infected quarters at the end of lactation. During the 2-year study, the incidence of clinical mastitis was 0.88 cases/cow-year; 32.2% were caused by streptococcal species other than Str agalactiae and 33.5% by gram-negative organisms. Among all new infections detected, 54.1% were caused by streptococcal species other than Str agalactiae and 25.7% by gram-negative bacteria. Among new infections, 41.6% occurred during the nonlactating period or within a few days of calving. Incidence of clinical mastitis was highest in the 1st month of lactation. Among 84 gram-negative infections, 42.8% were caused by Klebsiella pneumoniae, 20.2% by Escherichia coli, and 23.8% by Enterobacter spp. Among the many serotypes of K pneumoniae and E coli, none was predominant.     

Herd benefit-to-cost ratio and effects of a bovine mastitis control program that includes blitz treatment of Streptococcus agalactiae

Twelve dairy herds that had participated in the Pennsylvania Dairy Herd Improvement Association (DHIA) program for at least 12 months, that had a 12-month mean DHIA somatic cell count greater than 700,000 cells/ml, and that had greater than 25% of lactating cows infected with Streptococcus agalactiae participated in a herd blitz treatment program. Initially, quarter milk samples for bacteriologic culturing were collected from all lactating cows. Subsequently, all cows identified as infected with Str agalactiae were treated, using a commercial penicillin-novobiocin intramammary infusion product. In addition, a herd mastitis management program of postmilking teat dipping and treatment of all cows at the start of the nonlactating period was instituted. Thirty days after the initial herd visit, samples from all lactating cows were again cultured, and cows infected at that time were treated. Twelve months after the initial herd visit, samples from all lactating cows were again cultured. Mean prevalence of infection with Str agalactiae decreased (P less than 0.05) from 23.0% of quarters and 41.6% of cows initially to 3.4% of quarters and 9.3% of cows at 30 days and 1.6% of quarters and 4.2% of cows at 1 year. Mean herd DHIA somatic cell count decreased (P less than 0.05) from 918,000 cells/ml initially to 439,000 cells/ml at 30 days and 268,000 cells/ml at 1 year.(ABSTRACT TRUNCATED AT 250 WORDS)     

通用基因芯片对奶牛乳房炎主要致病茵的检测

应用生物信息学手段和查阅文献资料设计了金黄色葡萄球菌、大肠埃希菌、无乳链球菌、绿脓杆菌、停乳链球菌、乳房链球菌6种奶牛乳房炎主要致病菌的通用引物和金黄色葡萄球菌、大肠埃希菌、绿脓杆菌的寡核苷酸探针及无乳链球菌、停乳链球菌、乳房链球菌的特异引物,并用这3种特异引物扩增片段的纯化产物作为这3种链球菌的检测探针。在引物对样品中细菌的相应基因片段扩增的同时进行靶基因的生物素标记,扩增的产物与硝酸纤维素膜上的探针进行杂交,酶联、显色后根据芯片扫描仪的判读结果来确定奶牛乳房炎致病菌感染的种类。结果表明,建立的以16S rDNA为对象的基因芯片技术可以快速的检测出以上6种细菌,整个检测过程需要6h~7h,灵敏度高,特异性好,能快速的对奶牛乳房炎的主要致病菌做出诊断。     

Cheyletiella parasitivorax infestation of cats associated with skin lesions of man

Two dry-cow therapy products were evaluated in seven factory-supply dairy herds in the Waikato area. A product containing neomycin sulphate and the benzathine salt of penicillin (Neopen D.C. White; Smith-Biolab) was used in five herds, and one containing benzathine cloxacillin (Orbenin, Beecham) was used in two herds. Non-treated control cows were included in each herd. Both products were effective in eliminating intramammary infections with Staphylococcus aureus, Streptococcus uberis, and Streptococcus agalactiae. Efficacy of dry-cow therapy against S. aureus was 83.8% and 85.2% respectively. Spontaneous cure rate among controls was 30.8% for S. aureus during the dry period. Spontaneous cure rate for Str. uberis was 50%, while dry-cow therapy eliminated 100% and 77.8%, respectively, for the two products. Dry-cow therapy with either product eliminated more than 90% of Str. agalactiae infections while spontaneous cure rate was only 28.6%. These results further support the effectiveness of dry-cow therapy in reducing the level of subclinical mastitis in dairy herds by shortening the duration of intramammary infections.     

Prevalence of udder infections and mastitis in 50 California dairy herds

The California mastitis test (CMT) and bacteriologic culture were performed on samples of bulk-tank milk and cow-composite milk (n = 23,138 cows) from 50 California dairies, 19 of the 50 with known mastitis problems. Thirty-eight (76.0%) bulk-tank milk samples and 12,334 (53.3%) cows were positive by results of the CMT. Potential mastitis agents were isolated from 5,085 (22%) cows. Staphylococcus aureus was isolated from all 50 herds, Streptococcus agalactiae was isolated from 47 herds, and Mycoplasma sp was isolated from 24 herds. For cow-composite milk samples, the prevalences were 9.3% for Str agalactiae, 9.1% for S aureus, 0.9% for Mycoplasma sp, 1.2% for coliform bacteria, 0.9% for other streptococci, 0.8% for coagulase-negative staphylococci, and 1.3% for other organisms. The relative sensitivity and the relative specificity of the CMT performed on cow-composite milk samples were 83.4% and 55.2%, respectively, and the predictive value of positive CMT results was 34.2%.     

荧光定量PCR检测牛乳中金黄色葡萄球菌肠毒素A基因方法的建立

为建立检测牛乳中金黄色葡萄球菌(S.aureus)肠毒素A基因(SEA)定性定量的检测方法,本研究针对S.aureus SEA基因片段设计1对引物,将构建的重组质粒作为阳性对照,建立了S.aureus SEA DNA的SYBR Green I real-time PCR检测方法.结果显示,特异性产物Tm值为78.2℃～78.5℃,最低可检测到49.5 fg/μL(16.5拷贝)的阳性质粒.标准曲线的相关系数为0.99.与其他常见的产SEB的S.aureus、产SEC的S.aureus、无乳链球菌、大肠杆菌、嗜热链球菌、伤寒沙门氏菌、大肠杆菌DH5 α及JM109均无交叉反应.该检测方法具有较好的特异性和敏感性,为牛乳中S.aureus的快速检测提供了新的技术手段.     

Chelex-100法直接提取乳样中奶牛乳房炎主要致病菌DNA方法的建立

为建立一种直接从乳样中快速提取细菌DNA的方法,试验通过人工制备8个倍比稀释细菌的乳样,检测了Chelex-100法提取3种奶牛乳房炎主要致病菌(金黄色葡萄球菌、大肠杆菌和无乳链球菌)DNA进行PCR扩增的敏感性,并与苯酚—氯仿法进行了比较分析。结果显示,以Chelex-100法提取乳样中细菌DNA进行PCR扩增具有较高的敏感性,所检测金黄色葡萄球菌、大肠杆菌和无乳链球菌的最小浓度分别为10³、10²、10² CFU/mL;而使用苯酚—氯仿法提取乳样中各细菌DNA的PCR敏感性均为10⁴ CFU/mL。综上所述,Chelex-100法提取乳样细菌DNA的PCR敏感性可以满足临床检测奶牛乳房炎的需要,显现了简单快速、经济、无污染的优点,为从乳样中直接提取细菌DNA提供了新的思路,对PCR快速检测乳房炎致病菌具有重要意义。     

Antimicrobial resistance in streptococcal species isolated from bovine mammary glands

Streptococcal species isolated from dairy cows with clinical mastitis were obtained from mastitis research workers in Florida, Louisiana, New York, Vermont, Washington, and West Virginia. Seventy-one streptococcal isolates were tested, including 39 strains of Streptococcus agalactiae, 21 strains of S dysgalactiae, and 11 strains of S uberis. The minimal inhibitory concentration of erythromycin, lincomycin, oxytetracycline, penicillin, spectinomycin, streptomycin, and tetracycline was determined for each isolate. Differences were not detected among strains with respect to geographic origin. None of the strains was resistant to penicillin. Lincomycin was the next most effective antimicrobial, with only 2 resistant strains of each streptococcal species. There were no differences among the streptococcal species with respect to resistance to either penicillin or lincomycin. Streptococcus uberis was more likely to be resistant to erythromycin than were S agalactiae and S dysgalactiae (P less than 0.02). Streptococcus agalactiae and S uberis had similar distributions for resistance to oxytetracycline, tetracycline, spectinomycin, and streptomycin. Strains of S dysgalactiae were more likely to have intermediate resistance to oxytetracycline and streptomycin than were strains of S agalactiae and S uberis, which were highly resistant to oxytetracycline and streptomycin (P less than 0.001). Differences were not detected among the streptococcal species with respect to resistance to spectinomycin. Resistance to multiple antimicrobials was observed in all streptococcal species tested. Although S dysgalactiae appeared to have a greater percentage of strains (73%) that were resistant to multiple antimicrobials than did S agalactiae (31%) or S uberis (45%), differences were not statistically significant.