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1.
A total of 166 samples of secretions collected from Danish heifers suffering from clinically diagnosed summer mastitis were examined bacteriologically. One hundred and sixty three samples yielded positive findings whereas no growth was obtained from 3 specimens. The majority of samples revealed a mixed flora in which the predominant components were Actinomyces pyogenes, Peptostreptococcus indolicus, a microaerophilic coccus (Stuart-Schwan coccus), Fusobacterium necrophorum, Bacteriodes melaninogenicus and Streptococcus dysgalactiae. Pure cultures were recovered in only 7% of cases. P. indolicus was isolated from 875 of the cases, a microaerophilic coccus from 84%, A. pyogenes from 72%, Fusobacterium necrophorum from 52%, Str. dysgalactiae from 37%, B. melaninogenicus from 33% and various unidentified obligate anaerobic Gram-negative rods from 31%. The results confirm the complex bacterial aetiology of summer mastitis.  相似文献   

2.
A total of 143 secretions from clinical cases of summer mastitis (SM) in grazing cattle and from 89 cases of pyogenes mastitis (PM) in stabled cattle were examined bacteriologically. The typical bacteriological finding was a mixed flora in which the predominant organisms were Actinomyces pyogenes (SM-70%, PM-85%), Peptostreptococcus indolicus (54%, 54%), a microaerophilic coccus (Stuart-Schwan coccus) (26%, 25%), Fusobacterium necrophorum biovar B (22%, 12%), Bacteroides melaninogenicus (20%, 9%) and Streptococcus dysgalactiae (21%, 5%). All except six cases occurred in non-lactating animals or within three weeks after parturition. The majority of animals (about 90%) had only one quarter affected and no differences in quarter distribution were observed between the two groups.  相似文献   

3.
Mannitol agar for microbiologic diagnosis of bovine mastitis   总被引：1，自引：0，他引：1  
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.  相似文献   

4.
Using a simple in vitro test it was demonstrated that staphylococci, Streptococcus agalactiae, and micrococci, the species of bacteria which are commonly isolated from udder infections, adhered to mammary gland epithelial cells readily and in large numbers. Some strains of organisms which are associated with sporadic outbreaks or occur less commonly, like Str. dysgalactiae and Str. uberis, adhered moderately. Escherichia coli, Klebsiella spp., Corynebacterium pyogenes, C. bovis, Str. bovis, and Str. faecalis, species which are isolated occasionally, adhered poorly. From these studies, it appears that selective adherence of bacteria to the epithelial cells is a factor contributing to the ability of organisms to infect the mammary gland and may, therefore, be considered an important stage in the pathogenesis of bovine mastitis.  相似文献   

5.
With the aim of investigating the seasonal occurrence of Actinomyces pyogenes, Peptostreptococcus indolicus, Bacteroides melaninogenicus ss. levii and Fusobacterium necrophorum, and thus the potential for development of summer mastitis, clinically healthy Danish Holstein-Friesian heifers due to calve in the autumn were sampled from the teat tip, the conjunctiva and the oral cavity at 2-6 week intervals from 1979 to 1981. The overall isolation rates of F. necrophorum, P. indolicus and B. melaninogenicus ss. levii, in order of significance, were significantly higher during the pasture period whereas no differences in isolation rates of A. pyogenes between housed and pastured animals were detected. F. necrophorum was recovered almost exclusively from the oral cavity, P. indolicus and A. pyogenes occurred most frequently in samples from the teat skin, whereas isolates of B. melaninogenicus ss. levii were evenly distributed between conjunctiva and teat tip samples. A distinct seasonal pattern of the isolation rates of summer mastitis pathogens was recorded, which corresponded closely to the seasonal activity of symbovine insects, in particular the headfly Hydrotaea irritans (Fallén). However, the high proportion of clinically healthy bacterial carriers as compared with the incidence of clinical disease strongly suggests that as yet unknown contributing or triggering factors, apart from the mere presence of the relevant bacterial species, are required for the establishment and development of clinical summer mastitis.  相似文献   

6.
Forty-nine strains of microaerophilic gram-positive cocci (Stuart-Schwan cocci) isolated from summer mastitis, "pyogenes"-mastitis, other pyogenic conditions of Danish cattle and swine, and from the sheep headfly Hydrotaea irritans were biochemically characterized with the API 50 CH and API ZYM test kit systems, and screened for production of a variety of extracellular enzymes by agar plate methods. For comparison 4 strains isolated from Swedish cases of heifer and dry cow mastitis were included in the study. Similarity calculations indicated a high degree of homogeneity within the strains studied (similarity level 92%; group mean similarity 87%). The strains probably represent one species, although the taxonomic position of the organism remains unclear. The biochemical feature of the strains studied were very similar for strains isolated from cases of summer mastitis and strains from other sources of origin. It is suggested that the Stuart-Schwan coccus occurs as a natural cohabitant to Actinomyces pyogenes, Peptostreptococcus indolicus and the anaerobic organism characteristic of the bacterial complex isolated from summer mastitis and similar pyogenic conditions in ruminants and swine.  相似文献   

7.
The purpose of the study was to determine the etiological mastitis agents in cows from herds located in the western part of Poland in years 2003-2005. Bacteriological examinations of 18,713 samples taken from California Mastitis Test (CMT) positive quarters were performed according to standard methods. Some Gram-negative bacilli and Gram-positive cocci were examined with the use of API tests (Analytical Profile Index). It was found that 32.7% of samples that were CMT positive were culture negative and 9.3% of samples were contaminated. Streptococcus species (15.7%), coagulase negative staphylococci (CNS) (14.6%), Staph. aureus (8.6%), Gram-negative bacilli (4.0%) and Corynebacterium species (3.8%) were most frequently isolated. Escherichia coli (52.3%) dominated among Gram-negative bacilli followed by Klebsiella pneumoniae (4.1%), Pseudomonas aeruginosa (3.6%), Enterobacter cloacae (3.6%), Serratia marcescens (3.1%), Pasteurella multocida (3.1%), Acinetobacter lwofii (3.1%), and 26 other bacteria species. Stahylococcus xylosus (31.5%), Staph. hyicus (12.2%), Staph. haemolyticus (12.2%), Staph. sciuri (11.6%), Staph. chromogenes (8.8%), Staph. epidermidis (8.3%) and Staph. simulans (6.1%) were the most frequent CNS. Streptococcus uberis (50%), Str. dysgalactiae (19.7%), Str. acidominimus (6.6%) and Enterococcus faecalis (5.3%) were mostly found among CAMP-negative streptococci. An increase in frequency of mastitis caused by non-agalactiae streptococci, Staph. aureus, A. pyogenes and yeast-like fungi and a decrease in that produced by Str. agalactiae in 2005 as compared to years 2003 or 2004 were observed.  相似文献   

8.
A total of 1140 clinical cases of mastitis, with at least one inflamed quarter, were reported on 125 farms with somatic cell counts in bulk milk less than 150,000/ml. The average annual incidence was 17.9 cases per 100 cows and ranged from none to 80 cases per 100 cows. The microorganisms most frequently isolated were Escherichia coli (16.2 per cent), coagulase negative staphylococci (13.0 per cent), Staphylococcus aureus (9.6 per cent) and Streptococcus uberis (8.0 per cent). Only two cases of Streptococcus agalactiae were found. As the incidence of clinical mastitis increased, the proportion of S aureus also increased, while the proportions of E coli, S uberis and Streptococcus dysgalactiae remained about the same. Most of the clinical cases of mastitis occurred in early lactation, in November, December and January. However, after correction for the number of calvings per month, the incidence of mastitis was highest in the early summer months.  相似文献   

9.
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).  相似文献   

10.
奶牛乳腺炎病原菌的分离鉴定及耐药性分析   总被引：8，自引：1，他引：8  
2 0 0 2～ 2 0 0 3年采集上海地区 32个规模化奶牛场 433份奶样 ,共分离 2 54株细菌。其中包括金黄色葡萄球菌 48/2 54 (1 8 9% )、表皮葡萄球菌 34/2 54 (1 3 4% )、大肠杆菌 2 8/2 54 (1 1 0 % )、肺炎 /溶血巴氏杆菌 1 1 /2 54 (4 3 % )、无乳链球菌 2 5/2 54 (9 8% )、停乳链球菌 1 6/2 54(6 3 % )、乳房链球菌 1 5/2 54 (5 9% )及酵母菌 8/2 54 (5 9% )。表明葡萄球菌性乳腺炎仍占主导地位。药敏试验结果表明 ,分离菌株对临床常用的多种抗菌药物都产生了耐药性 ,耐药菌株的出现和抗生素的残留给乳腺炎的防治及食品安全带来了隐患  相似文献   

11.
Accurate identification of mastitis pathogens is often compromised when using conventional culture-based methods. Here, we report a novel, rapid assay tested for speciation of bacterial mastitis pathogens using high-resolution melt analysis (HRMA) of 16S rDNA sequences. Real-time PCR amplification of 16S rRNA gene fragment, spanning the variable region V5 and V6 was performed with a resulting amplicon of 290bp. First, a library was generated of melt curves of 9 common pathogens that are implicated in bovine mastitis. Six of the isolates, Escherichia coli, Streptococcus agalactiae, Klebsiella pneumoniae, Streptococcus uberis, Staphylococcus aureus and Mycoplasma bovis, were type strains while the other 3, Arcanobacterium pyogenes, Corynebacterium bovis and Streptococcus dysgalactiae, were bovine mastitis field isolates. Four of the type strains, E. coli, S. agalactiae, K. pneumoniae and S. aureus, were found to be of human origin, while the other 3 type strains were isolated from bovine infections. Secondly, the melt curves and corresponding amplicon sequences of A. pyogenes, E. coli, S. agalactiae, S. dysgalactiae, K. pneumoniae, S. uberis and S. aureus were compared with 10 bovine mastitis field isolates of each pathogen. Based on the distinct differences in melt curves and sequences between human and bovine isolates of E. coli and K. pneumoniae, it was deemed necessary to select a set of bovine strains for these pathogens to be used as reference strains in the HRMA. Next, the HRMA was validated by three interpreters analyzing the differential clustering pattern of melt curves of 60 bacterial cultures obtained from mastitis milk samples. The three test interpreters were blinded to the culture and sequencing results of the isolates. Overall accuracy of the validation assay was 95% as there was difficulty in identifying the streptococci due to heterogeneity observed in the PCR amplicons of S. uberis. The present study revealed that broad-range real-time PCR with HRMA can be used as a powerful, fast and low-cost tool for the differentiation of clinically important bacterial mastitis pathogens.  相似文献   

12.
Pc. indolicus was isolated from 42.7% and Cb. pyogenes from 48.7% of 150 pus-samples from abscesses in pigs (slaughter-house material, Table I). In 41 specimens the two organisms were found together. Further, Pc. indolicus was demonstrated in 22.4% of 290 swabs from apparently normal skin and mucous membranes of piglets (autopsy material, Table II). By gel diffusion analysis the strains of Pc. indolicus were referred to the serotypes B, C, D, E, or F. The type distribution (Table III) possibly reflects a type-related variation in the invasive properties of Pc. indolicus.  相似文献   

13.
A total of 1050 samples from apparently healthy cattle were examined bacteriologically with special regard to Pc. indolicus and Cb. pyogenes.Pc. indolicus was found in 58 % of 130 samples from tonsils (slaughterhouse material), in 23 % of 620 samples from the vagina of cows, in 22 % of 100 samples from the vagina of calves and heifers, in 5 % of 100 samples from the conjunctival sac of cows, and in 10 % of 100 samples from the nasal cavity of cows (Table 1). Cb. pyogenes was found in 51 %, 17 %, 19 %, 8 %, and 6 %, respectively. Both organisms were found in each of 9 herds examined, though with varying frequency (Tables 2, 3, and 4).Altogether Pc, indolicus was found in 254 (24 %) and Cb. pyogenes in 205 (20 %) of the samples examined (Table 1). In 127 samples both organisms were present. Eleven of the strains of Pc. indolicus were β-hemolytic, the rest non-hemolytic.By gel diffusion analysis the strains of Pc. indolicus as well as those of Gb. pyogenes could be identified with strains originating from pathological conditions in cattle. With Serotype B occurring most frequently, usually two or three different types of Pc. indolicus were found in each of the herds examined (Tables 5, 6, and 7).The investigation has shown that Pc. indolicus is widespread among healthy cattle, and given evidence to suggest that Pc. indolicus and Cb. pyogenes are natural cohabitants.  相似文献   

14.
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.  相似文献   

15.
Traditional microbiological methods for identification of Streptococcus spp. causing bovine mastitis have been demonstrated to be less than highly reliable. PCR-RFLP analysis of 16S-23S ribosomal DNA was used to characterise seven reference strains of streptococcal mastitis pathogens as well as four reference strains of other gram-positive, catalase-negative cocci of bovine origin to allow comparative identification of field isolates. RFLP analysis of PCR products, using a combination of two restriction endonucleases in single reactions (HaeIII and AluI, HaeIII and RsaI or AluI and RsaI) generated unique patterns for species of Streptococcus, Enterococcus and Lactococcus. One hundred field isolates of Streptococcus spp. collected from cows with clinical or subclinical mastitis were tested. Fifty-seven isolates, classified by conventional tests as S. uberis, were identified as 47 S. uberis and six S. parauberis by their unique RFLP patterns. The remaining four isolates had RFLP patterns distinct from the reference strains and two of these were identified as closely related to S. iniae and two to Aerococcus viridans by 16S rRNA sequencing. Conventional identification of 17 S. agalactiae and 26 S. dysgalactiae subsp. dysgalactiae was confirmed by RFLP. Use of a combination of restriction enzymes in a single tube enabled the rapid, accurate, cost effective and easily performed identification of all major streptococcal mastitis pathogens.  相似文献   

16.
Streptococcus uberis, as one of the principal causes of bovine streptococcal mastitis, has been characterized serologically and biochemically. Serological grouping of S. uberis revealed polysaccharide antigens of groups E, G, P and U. The biochemical properties of S. uberis, determined with the Strep-Zym identification system, differed clearly from those of S. agalactiae and S. dysgalactiae. Some cultures of S. uberis produced the enzymes hyaluronidase and neuraminidase. In addition S. uberis partly demonstrated CAMP-like synergistic hemolytic activities on sheep blood agar, reacted specifically with the lectins from Helix pomatia and Dolichos biflorus and produced bacteriocin-like inhibitors. This reactions, possibly of importance as virulence factors, as well as "DNA-fingerprinting" of S. uberis, might serve as individual markers of the respective cultures in epidemiological studies.  相似文献   

17.
The antibacterial properties of bacteriophage lytic enzymes may be of importance in future mastitis control programs. A prophage was isolated from a strain of Streptococcus uberis (ATCC 700407) following exposure to mitomycin C. Partial sequencing of the phage DNA revealed a putative lysin based on sequence similarity to other streptococcal phage lysins. The putative lysin (Ply700) was recombinantly expressed in Escherichia coli, and chromatographically purified. Addition of the purified Ply700 to bacterial suspensions of S. uberis, Streptococcus pyogenes, and Streptococcus dysgalactiae caused a rapid, calcium-dependent lysis while there was little activity against Streptococcus agalactiae, Staphylococcus aureus, or E. coli. Killing of S. uberis in milk by Ply700 (50 microg/ml) was confirmed by plate count assay. Activity was related to the initial concentration of bacteria in that 31% killing (P<0.05) was observed with an inoculating dose of approximately 4500 cfu/ml, while 81% killing (P<0.01) was observed when the inoculum was reduced to approximately 600 cfu/ml. In contrast, complete sterilization was observed in parallel cultures suspended in assay buffer indicating that factors in milk are able to neutralize the lysin. Functional characterization of the C-terminal domain, as a component of a GFP fusion protein, revealed its calcium-dependent ability to bind to S. uberis. The C-terminal domain may have utility in targeting S. uberis while it remains to be determined if the lysin by itself has sufficient potency in milk for effective use in the control of S. uberis mastitis.  相似文献   

18.
对昆明地区多个奶牛场的奶牛临床型乳房炎病原菌进行分离、鉴定,从114份乳样中共分离出细菌13种305株,经培养特性、染色镜检、生化鉴定等,鉴定出金黄色葡萄球菌63株,占20.66%;表皮葡萄球菌26株,占8.52%;无乳链球菌53株,占17.38%;停乳链球菌23株,占7.54%;乳房链球菌19株,占6.23%;乳链球菌14株,占4.59%;粪链球菌15株,占4.92%;化脓链球菌14株,占4.59%;大肠杆菌48株,占15.74%;真菌12株,占3.93%。分别对分离菌株进行17种药物的敏感性试验,除真菌外,所有菌株对庆大霉素、新霉素、环丙沙星、诺氟沙星、林可霉素、乳肿康均高度敏感,对临床常用的青霉素、链霉素、鱼腥草等具有较强的耐药性。  相似文献   

19.
Twenty-nine dairy farms were selected to determine the incidence of clinical mastitis, prevalence of sub-clinical mastitis and bacterial aetiology in the West Littoral Region of Uruguay. In samples taken by the owner and frozen at -20 degrees C during a week the incidence rate of clinical mastitis was determined as 1.2 cases per 100 cow-months at risk. Staphylococcus aureus was the most common isolated pathogen in 37.5% of 40 milk samples from clinical cases obtained in 1 month. No bacteria grew in the 32.5% of the total samples. A sub-sample including 1077 dairy cows from randomly selected farms was used to determine the prevalence of sub-clinical mastitis. These samples were taken on one visit to each farm. The prevalence was 52.4% on a cow basis and 26.7% on an udder quarter basis. In 55.1% of the quarters of the selected animals with more than 300,000 cells/ml there was no growth. The isolated pathogens from sub-clinical cases and their relative frequencies were: Staphylococcus aureus 62.8%, Streptococcus agalactiae 11.3%, Enterococcus sp. 8%, coagulase-negative staphylococci 7.4%, Streptococus uberis 6.4%, Streptococcus dysgalactiae 1.8%, Escherichia coli 1.5% and Staphylococcus hyicus coagulase-positive 0.6%.  相似文献   

20.
We previously showed that Staphylococcus aureus cells adhered mainly to an elongated cell type, present in cultures of bovine mammary gland cells. Moreover, we showed that this adhesion was mediated by binding to fibronectin. The same in vitro model was used here, to study adhesion of other important mastitis pathogens. Like the S. aureus strains, the Streptococcus dysgalactiae strains adhered mainly to elongated cells, which seemed to be mediated by fibronectin binding. In contrast, Streptococcus uberis strains adhered mainly to cubic cells. Since the cubic cells did not express fibronectin and S. uberis cells bound fibronectin less efficiently, the adhesion of S. uberis cells was independent of fibronectin binding. Streptococcus agalactiae strains adhered poorly to both cell types. The specificity and efficiency of adhesion of Escherichia coli strains was strongly strain dependent. None of the S. agalactiae and E. coli strains tested was able to bind fibronectin efficiently. The results suggest that the different mastitis pathogens have different target cell specificities and use different mechanisms to adhere to cells of the bovine mammary gland.  相似文献   

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