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1.

Background

Methicillin-resistant staphylococci (MRS) are increasingly being isolated in bovine mastitis. The aim of our study was to evaluate the occurrence of MRS in Finnish mastitis milk samples and characterize the MRS isolates using molecular methods.

Results

Methicillin-resistant S. aureus (MRSA) was a rare finding in bovine mastitis in Finland. Only two out of 135 (1.5%) S. aureus isolates were positive for mec genes. One of these carried mecA and was of spa type t172, SCCmec type IV and ST375, and the other harboured mecC, being spa type t3256, and ST130. MRSA ST375 is common among human MRSA isolates in Finland, but this is the first report in the country of bovine mecC MRSA. In coagulase-negative staphylococci (CoNS) originating from bovine mastitis, methicillin resistance was more common. In the two CoNS collections studied, 5.2% (17/324) and 1.8% (2/110) of the isolates were mecA positive. Eighteen of these were methicillin-resistant S. epidermidis (MRSE), which were divided into 6 separate PFGE clusters. One pulsotype was detected in different parts of the country, indicating clonal spread. Most MRSE (13/18) were of SCCmec type IV, one was of type V and four were non-typeable. Comparison with a human staphylococcal database indicated that bovine MRSE strains were not closely related to human MRSE isolates.

Conclusions

The occurrence of MRS, especially MRSA, in bovine mastitis in Finland was low. Most methicillin-resistant bovine CoNS are MRSE, and we found evidence of a bovine MRSE strain that may spread clonally. This is the first report of a Finnish bovine isolate of MRSAmecC ST130. The study provides a baseline for further MRS monitoring.  相似文献   

2.
The objective of this study was to determine the frequency of different mastitis pathogens in 1255 milk samples collected from 368 lactating cows on 24 dairy farms in Korea during 2008. The proportion of cows and quarter milk samples having SCC ? 200,000 cells/ml, an indicator of udder infection, was 54.3% (200/368 cows) and 35.5% (446/1255 samples), respectively. Of the 446 milk samples subjected to bacteriological examination, 16.5% (74) showed no bacterial growth and 3.5% (16) were contaminated. In total, 356 of 1255 (28.3%) samples were bacteriologically positive, from which 415 bacteria were isolated. The most frequently isolated pathogen was coagulase-negative staphylococci (40.7%), followed by Gram-negative bacteria (19.5%) other than Escherichia coli, Staphylococcus aureus (12.2%), Streptococcus uberis (5.3%), Enterococcus spp. (4.8%), E. coli (4.5%), and environmental streptococci (3.1%) other than S. uberis. This study demonstrates that environmental pathogens were the vast majority of bacteria isolated from mastitic bovine milk samples in Korea.  相似文献   

3.
Summary

The results of a mastitis control field experiment in Utrecht over three and a half years, including sevens herds with a total of 225 lactating cows are presented. Every case of clinical mastitis was examined bacteriologically. Quarter samples were taken routinely at an average interval of five weeks.

The incidence of clinical mastitis in each herd at the start of the experiment varied from 10 to 104 quarter cases per 100 cows per year. The majority of cases of clinical mastitis (33.1 per cent) occurred during the first month of lactation.

In 74.4 per cent of the lactations marked by clinical mastitis only a single case was observed during that lactation period. If the animals were on pasture during the first month of lactation, the incidence of clinical mastitis was significantly lower during this month, compared with the first month when they were housed. Str. dysgalactiae was the most common isolate from clinical cases (21.5 per cent). A total proportion of 40.3 per cent of the clinical cases were caused by cocci, 20.3 per cent by Gram‐negative bacteria and 16.0 per cent were bacteriologically negative. Of the clinical cases caused by streptococci and staphylococci, 33.3 per cent were preceded by subclinical infection, compared with 11.8 per cent of the clinical cases due to E. coli. The overall incidence of clinical mastitis in this experiment decreased from 47 to 31 per 100 cows per year.  相似文献   

4.
The prevalence of bovine mastitis has been reduced over the past 25 years due to the implementation of a five-point control plan aimed at reducing exposure, duration and transmission of intramammary infections by bacteria. This has markedly reduced the incidence of bovine mastitis caused by bacteria which show a contagious route of transmission, but has had little effect on the incidence of mastitis due to bacteria which infect the gland from an environmental reservoir. Streptococcus uberis is one such bacterium which is responsible for a significant proportion of clinical mastitis worldwide.The inadequacies of the current methods of mastitis control have led to the search for additional measures, particularly vaccines to prevent intramammary infection by this bacterium. Such an approach requires detailed knowledge of the pathogenesis of intramammary infection. Our understanding of this area has grown in recent years but a lack of information still hampers disease control. Both live vaccines and, recently, crude sub-unit vaccines have shown promise against bovine mastitis due to S. uberis. Vaccines against mastitis must, however, be able to control infection without the participation of a marked inflammatory response. This review provides an overview of the recent advances which have been made in our understanding of host-pathogen interactions which promote infection and disease and highlights areas for strategic research aimed at controlling this bacterial infection.  相似文献   

5.
《Veterinary microbiology》1988,17(4):357-365
Isolates of bacteria from normal teats were used to attempt colonization of teats of dry cows or neonatal calves. Isolates for inoculation were chosen on the basis of ability to inhibit mastitis pathogens in vitro, with the ultimate goal of in vivo inhibition of mastitis pathogens at the teat surface. Three bacterial normal flora isolates (Corynebacterium xerosis, Bacillus sp. and Aerococcus viridans) persisted less than 10 days on the teats of dry cows. The fourth isolate, Staphylococcus hominis 1, was studied in greatest detail because studies characterizing the normal teat flora showed staphylococci to be the predominant flora. The S. hominis 1 isolated used for inoculation was an inhibitor of Gram-positive mastitis pathogens. It was a biotype not found on these teats prior to inoculation, thus facilitating identification of the inoculated isolate on sequential sampling. Colonization of newborn calves, before other bacterial floral became established, resulted in recovery of inoculated S. hominis 1 for an average of 51 days or longer. On dry cow teats it was detected for up to 28 days. On several occasions the inoculated S. hominis 1 was found in pure culture. Since many new infections occur during the dry period, the colonization of dry cow tests with S. hominis 1 organisms inhibitory for Gram-positive pathogens should be tested as an adjunct to other methods of mastitis prevention.  相似文献   

6.
Staphylococcus aureus is internationally recognised as a principal agent of mastitis and the foremost reason for economic loss in the dairy industry. The limited data available on organism-specific antibiotic resistance surveillance in dairy cattle have stimulated the need for such a review article. The objective of this study was to review relevant literature on antimicrobial resistance of mastitis-causing staphylococci isolated from dairy cows in South Africa compared to other countries. Factors relating to the incidence of mastitis and treatment strategies in terms of the One Health concept and food security were included. The Web of Science (all databases) and relevant websites were used, and articles not written in English were excluded. The incidence of mastitis varied between South Africa and other countries. Antimicrobial resistance patterns caused by S. aureus also varied in regions within Southern Africa and those of other countries although some similarities were shown. Antimicrobial resistance differed between S. aureus bacteria that were maltose positive and negative (an emerging pathogen). The results highlighted the importance of the availability of organism-specific surveillance data of the incidence of mastitis and antibiotic resistance for specific countries and within similar climatic conditions. Accurate knowledge about whether a specific pathogen is resistant to an antibiotic within a certain climate, country, area or farm should reduce the incidence of unnecessary or incorrect treatment with antibiotics. This should enable dairy farmers to deal with these organisms in a more effective manner. Therefore such research should be ongoing.  相似文献   

7.
The anaerobic mastitis incidence was used to study the bovine udder response in anaerobic bacterial mastitis caused by the Gram-positive bacterial strain of Clostridium perfringens. Milk samples positive for C. perfringens were assayed for NO and lysozyme. The model produced a strong NO and lysozyme response which correlated positively with the severity and outcome of the disease (subclinical and clinical stages). This study is, to our knowledge, the first to suggest a possible link between NO and lysozyme and bovine mastitis caused by C. perfringens. The results raise the possibility that interfering with NO production during mastitis may help to prevent tissue damage.  相似文献   

8.
Natural killer (NK) cells are early responders in bacterial infections but their role in bovine mastitis has not been characterized. For the first time, we show the presence of NK cells (NKp46+/CD3−) in bovine mammary gland tissue after an intramammary challenge with Escherichia (E.) coli. A small number of NK cells was detected in milk from quarters before and during an E. coli challenge. In vitro cultures of primary bovine mammary gland epithelial cells stimulated with UV irradiated E. coli induced significant migration of peripheral blood NK cells (pbNK) within 2 h. Furthermore, pbNK cells significantly reduced counts of live E. coli in vitro within 2 h of culture. The results show that bovine NK cells have the capacity to migrate to the site of infection and produce antibacterial mediators. These findings introduce NK cells as a leukocyte population in the mammary gland with potential functions in the innate immune response in bovine mastitis.  相似文献   

9.
This study was carried out to determine the prevalence of coagulase-negative staphylococci in clinical and subclinical mastitis in commercial and small-scale farms in Zimbabwe. Thirty five quarter milk samples from clinical mastitis cases and 371 quarter milk samples from cows with subclinical mastitis were cultured for bacterial pathogens. The most frequent pathogens isolated in clinical mastitis were the enteric bacteria (31.4%), followed by coagulase negative staphylococci (22.9%) and then Staphylococcus aureus (17.1%), whereas in subclinical mastitis S. aureus (34.2%) and coagulase-negative staphylococci were (33.2%) the most common. Bacillus species were only isolated in milk samples from subclinical mastitis. Coagulase-negative staphylococci were observed in mixed infections with other bacteria in only 2.2 of the 406 milk samples from clinical and subclinical mastitis where they were isolated together with Bacillus species in 6 of the 9 mixed infection cases. About 95% of the milk samples from which 131 coagulase-negative staphylococci were isolated had correspondingly high somatic cell counts. The coagulase-negative staphylococci isolated most frequently were S. chromogenes (7.9%), S. epidermidis (7.4%) and S. hominis (5.9%). They were all associated with high somatic cell counts. All the coagulase-negative staphylococci isolates were susceptible to cloxacillin and erythromycin, and more than 90% of the isolates were susceptible to neomycin, penicillin and streptomycin. The highest resistance was to tetracycline (17.6%), followed by lincomycin (13.7%). About 8% of the isolates were resistant to both penicillin and streptomycin.  相似文献   

10.
The effect of intramammary injection of recombinant bovine interleukin‐8 (rbIL‐8, 1 mg/10 ml of saline) on quarter milk levels of somatic cell count (SCC), chemiluminescence (CL) activity and counts of total bacteria and Staphylococcus aureus (S. aureus) was investigated, using 10 Holstein cows with an early stage or a late stage of subclinical mastitis naturally infected with S. aureus. In the late‐stage group, milk SCC and CL activity had significant rises with maximum levels at 6 h, following maintained high levels thereafter post‐cytokine injection. The counts in milk total bacteria and S. aureus were insignificantly decreased, being increased back on day 7 post‐cytokine injection. Thus, the cytokine was inefficient for the late‐stage subclinical mastitis. However, in the early‐stage group milk SCC and CL activity declined to under pre‐injection levels on day 7 after marked and significant rises at 6 h and day 1 post‐cytokine injection. The milk total bacterial count decreased significantly on days 0.25 and 2. Furthermore, the milk S. aureus count was decreased significantly on days 1, 2, 3 and 7 by the cytokine injection. These results suggest that the rbIL‐8 has a potential as a therapeutic agent of the subclinical mastitis of dairy cows, if the cytokine is applied at an initial stage of infection.  相似文献   

11.
A cross-sectional study was conducted between January and July 2014 in Unguja island of Zanzibar to establish prevalence of subclinical mastitis (SCM) in smallholder dairy cows and patterns of antibacterial susceptibility of major mastitis pathogens isolated. A total of 416 dairy cows from 201 farmers were randomly selected from three districts of Unguja Island to participate in the study. Questionnaire interview, field observation, individual cow examination, California Mastitis Test (CMT) and bacteriological examination were carried out. Kirby-Bauer disc diffusion technique was used to test drug sensitivity for common bacteria isolated. Based on CMT results, the overall prevalence of SCM was 28.6, 48.8 and 64.7% at quarter, cow and farm level, respectively. Prevalence of bacterial infection was recorded at 42.9, 70.9 and 78.6% at quarter, cow and farm examined, respectively. The common bacteria isolated included Staphylococcus aureus (36.8%), Pseudomonas aeruginosa (17.8%), Staphylococcus epidermidis (16.1%), Klebsiella spp. (9.5%), Micrococcus spp. (6.3%) and Escherichia coli (4.9%). In conclusion, findings of this study demonstrated high level of subclinical mastitis at farms, cows and quarters levels with both contagious and environmental bacterial pathogen involved. Therefore, efforts should be directed to the decreased subclinical mastitis by improving sanitary measures and proper milking practice.  相似文献   

12.
Mastitis, the inflammation of the mammary gland, is an important disease affecting dairy animals worldwide. The disease is caused by mammary pathogenic bacteria and Escherichia coli are frequently implicated. Virulence factors of mammary pathogenic E. coli are only partially known and intramammary challenge with LPS elicits neutrophil recruitment in experimental bovine and murine mastitis models. We have previously shown that neutrophil recruitment in LPS-induced murine mastitis is strictly dependent on mammary alveolar macrophages. However, the relative role of alveolar macrophages and blood neutrophils in E. coli mastitis is not well defined. To this end, we selectively depleted mammary alveolar macrophages or blood neutrophils before intramammary challenge with E. coli strain P4 (ECP4). Mice depleted of alveolar macrophages prior to intramammary challenge recruited neutrophils normally and restricted bacterial growth and interstitial invasion. Importantly however, upon depletion of alveolar macrophages, ECP4 invaded the mammary alveolar epithelial cells and formed intracellular bacterial communities. In contrast, neutrophil depletion prior to intramammary infection with ECP4 was associated with unrestricted bacterial growth, tissue damage, severe sepsis and mortality. This study suggests that neutrophils but not alveolar macrophages provide essential antimicrobial defense against mammary pathogenic E. coli. Furthermore, we show here similar invasion after depletion of alveolar macrophages as in our previous studies showing that LPS/TLR4 signaling on alveolar macrophages abrogates ECP4 invasion of the mammary epithelium. Interestingly, similar ECP4 invasion and formation of intracellular communities were also observed following intramammary infection of either iNOS gene-deficient or IL-1 receptor type 1 gene-deficient mice.  相似文献   

13.
The multidrug efflux system in bacteria can reduce antibiotic concentration inside the cell, leading to failure in the treatment of bacterial diseases. This study evaluated the influence of two efflux pump inhibitors (EPIs), phenylalanine arginyl ß-naphthylamide (PAßN) and 1-(1-naphthylmethyl)-piperazine (NMP), on the gene expression of three multidrug efflux systems, AcrAB, AcrEF and EmrAB in Escherichia coli bovine mastitis isolates resistant to ampicillin and sulfamethoxazole/trimethoprim simultaneously. Each isolate had at least three multidrug efflux system genes. The acrA and acrB had the lowest expression levels in all treatments, while the emrA or emrB showed the highest expression levels in the presence of ampicillin, sulfamethoxazole/trimethoprim, PAβN and NMP. EPIs also contributed to the decrease in arcF expression when used in combination with ampicillin treatment. Since PAßN showed stronger effects than NMP, it may serve as an alternative to assist in the antimicrobial therapy of mastitis.  相似文献   

14.
Development of an appropriate Staphylococcus aureus vaccine for bovine mastitis has eluded researchers for decades. The ability of S. aureus to form a protective exopolysaccharide capsule has posed a major obstacle because of the multiple serotypes and the poor immune response elicited by exopolysaccharides. This study characterized S. aureus serotypes isolated from cases of bovine mastitis obtained from veterinary diagnostic laboratories that service 44% of the dairy cattle in the United States. Major milk producing areas of the northeast, north central, Pacific coast and southwest were proportionately represented. Sub-samples of mastitic milk that contained S. aureus were frozen and sent to our laboratory for strain serotyping. The only other regional serotyping of S. aureus from bovine mastitis to date was done in France. The primary serotypes found were types 5 (51%) and 8 (18%) and 31% were non-typeable. In the current study, serotype 5 accounted for 18% of the isolates and serotype 8 for 23%. More importantly 59% of the isolates were not typeable with either type 5 or 8 antisera. These data indicate that S. aureus vaccines employing serotypes 5 and 8 would only be marginally effective in the United States. These data also suggest that development of a S. aureus vaccine for bovine mastitis should take into account regional variation in S. aureus serotypes.  相似文献   

15.
Staphylococcus spp. are the single most important cause of mastitis in dairy cows. Aiming for a rapid and sensitive method for detection of staphylococci, a magnetic bead-based enzyme-linked immunosorbent assay (ELISA) employing MAb for the detection of staphylococci in milk was developed. Pretreatment of the sample with lysostaphin increased the sensitivity of the assay. The method is specific for most of staphylococci which cause mastitis in bovines, detects between 104–105 organisms per ml and takes three hours. Thus, the principle for staphylococci ELISA based on monodisperse magnetic beads is a way for rapid detection of staphylococci in milk and possibly also for detecting staphylococci by direct testing of other clinical specimens. The system may be automated.  相似文献   

16.
Escherichia coli is a major pathogen in the aetiology of bovine mastitis. Although classically considered to be an environmental pathogen causing mainly transient infection, the incidence of persistent E. coli mastitis infections may be increasing, suggesting an adaptation of this pathogen to the bovine udder environment. Mastitis E. coli strains have been demonstrated to enter bovine mammary cells in vitro but little is known about the invasion mechanism or the intracellular fate of the bacteria. In order to further understand the pathogenesis of persistent E. coli bovine mastitis we investigated the intracellular trafficking of mastitis E. coli isolates in primary bovine mammary cells using confocal microscopy and fluorescent markers of endocytic compartments. Consistent with other studies, mastitis E. coli were found to invade primary bovine mammary cells in vitro. This process did not involve in the rearrangement of the actin cytoskeleton. Intracellular bacteria were observed within membrane-bound compartments that labelled with the early endosomal marker phosphatidylinositol 3-phosphate (PtdIns(3)P) and also within late endosome-like compartments labelled with the small GTPase Rab7, indicating an endocytic mechanism of bacterial internalization. Bacteria were not observed within acidified lysosomal compartments or autophagic vacuoles, suggesting that the internalized bacteria are not targeted for lysosomal degradation via either the classical endocytic pathway or the autophagic response. Our findings are consistent with an endosomal survival niche for the internalized bacteria, allowing them to evade host immune responses and establish an infection reservoir that could later re-emerge as a recurrent clinical mastitis episode.  相似文献   

17.
We conducted a prospective observational study on clinical and subclinical mastitis in 30 commercial meat-producing sheep flocks from 2 regions of the province of Quebec, Canada. A total of 2792 ewes selected in late gestation were followed from lambing to weaning of lambs. The incidence of clinical mastitis for the total lactation period (average of 58 days) ranged among flocks from 0 to 6.6%, with a median of 1.2%. The most frequently isolated bacteria from the cases of clinical mastitis, in pure or mixed culture, were Mannheimia haemolytica (26%), Staphylococcus aureus (23%), and coagulase-negative staphylococci (17%). Incidence of clinical mastitis was higher in ewes that gave birth to 3 or more lambs and from the Estrie region, and was associated with an increase in ewe mortality, an increase in lamb mortality at the litter level, and a decrease in lamb's weaning weight for lambs born in multiple litter size or from ewes ≥4 years old.Among 354 selected ewes with clinically normal udder at the end of lactation, 28.8% had potentially pathogenic bacteria isolated from milk. The most prevalent bacteria were S. aureus (9.3%) and coagulase-negative staphylococci (9.3%). The risk of having a positive culture in at least one half was different between the two regions. Prevalence of ewes (n = 261) with California Mastitis Test (CMT) positive result in at least one half was 24.1 and 14.9% using a cut-off of ≥1+ and ≥2+, respectively. Prevalence of culture-positive udder halves was 11.7% for CMT-negative compared with 53.6% for CMT 3+ halves. CMT status was positively associated with the isolation of coagulase-negative staphylococci, M. haemolytica, S. aureus, and various Streptococcus species, but not with other isolated bacteria. Additionally, prevalence of CMT-positive halves was higher in ewes from the Estrie region, aged of ≥4 years versus 1 year, having clinical mastitis previously detected in the lactation and/or with low body condition score. Lamb weaning weight was associated with CMT status of ewes, while weaning weight was not associated with milk culture results. More research is needed to understand the dynamic of milk SCC and IMI in ewes from meat-producing flocks, its economical impact and best ways to control it.  相似文献   

18.

Background

Mastitis is a high incidence disease in dairy cows. The acute stage is considered painful and inflammation can lead to hyperalgesia and thereby contribute to decreased welfare. The aim of this study was to examine changes in nociceptive responses toward cutaneous nociceptive laser stimulation (NLS) in dairy cows with experimentally induced Escherichia coli mastitis, and correlate behavioral changes in nociceptive responses to clinical and paraclinical variables.

Methods

Seven Danish Holstein-Friesian cows were kept in tie-stalls, where the E. coli associated mastitis was induced and laser stimulations were conducted. Measurements of rectal temperature, somatic cell counts, white blood cell counts and E. coli counts were conducted. Furthermore, scores were given for anorexia, local udder inflammation and milk appearance to quantify the local and systemic disease response. In order to quantify the nociceptive threshold, behavioral responses toward cutaneous NLS applied to six skin areas at the tarsus/metatarsus and udder hind quarters were registered at evening milking on day 0 (control) and days 1, 2, 3, 6 and 10 after experimental induction of mastitis.

Results

All clinical and paraclinical variables were affected by the induced mastitis. All cows were clinically ill on days 1 and 2. The cows responded behaviorally toward the NLS. For hind leg stimulation, the proportion of cows responding by stepping was higher on day 0 than days 3 and 6, and the frequency of leg movements after laser stimulation tended to decrease on day 1 compared to the other days. After udder stimulation, the proportion of cows responding by stepping was higher on day 1 than on all other days of testing. Significant correlations between the clinical and paraclinical variables of disease and the behavioral responses toward nociceptive stimulation were found.

Conclusions

Changes in behavioral responses coincide with peaks in local and systemic signs of E. coli mastitis. During the acute stage of E. coli mastitis nociceptive thermal stimulation on hind leg and mammary glands results in decreased behavioral responses toward nociceptive stimulation, which might be interpreted as hypoalgesia.  相似文献   

19.
The global dairy industry, the predominant pathogens causing mastitis, our understanding of mastitis pathogens and the host response to intramammary infection are changing rapidly. This paper aims to discuss changes in each of these aspects. Globalisation, energy demands, human population growth and climate change all affect the dairy industry. In many western countries, control programs for contagious mastitis have been in place for decades, resulting in a decrease in occurrence of Streptococcus agalactiae and Staphylococcus aureus mastitis and an increase in the relative impact of Streptococcus uberis and Escherichia coli mastitis. In some countries, Klebsiella spp. or Streptococcus dysgalactiae are appearing as important causes of mastitis. Differences between countries in legislation, veterinary and laboratory services and farmers' management practices affect the distribution and impact of mastitis pathogens. For pathogens that have traditionally been categorised as contagious, strain adaptation to human and bovine hosts has been recognised. For pathogens that are often categorised as environmental, strains causing transient and chronic infections are distinguished. The genetic basis underlying host adaptation and mechanisms of infection is being unravelled. Genomic information on pathogens and their hosts and improved knowledge of the host's innate and acquired immune responses to intramammary infections provide opportunities to expand our understanding of bovine mastitis. These developments will undoubtedly contribute to novel approaches to mastitis diagnostics and control.  相似文献   

20.
Six hundred and thirty clinically-normal milk samples from dairy goat flocks comprising a mixed population of German Alpine, Toggenburg, Saanen and Galla crosses were examined over a 3-month period to determine the prevalence of bacterial organisms. Bacteria were isolated in 28.7% of the milk samples (181/630) either singly (92.8%) or in combination (7.2%). The most prevalent bacterial organisms were Staphylococcus spp. (60.3%), followed by Micrococcus spp. (17.7%), Acinetobacter spp. (5%), Actinomyces spp. (5%) and Streptococcus spp. (1.1%). The Staphylococcus spp. were mainly coagulase negative (64.3%). Coagulase-negative staphylococci and coagulase-positive staphylococci accounted for 37.5% and 22.7% respectively of the total bacteria isolated. The isolation of bacteria, some of which are important in clinical and subclinical mastitis, in apparently normal caprine milk, indicates that particular attention should be given to the management of these dairy goat flocks in order to avoid the development of cases of clinical mastitis.  相似文献   

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