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1.
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   

2.
The objective was to determine if induced mammary inflammation at cessation of milking influenced growth of gram-positive mastitis pathogens in mammary secretions, particularly during early involution. Growth of all mastitis pathogens evaluated was similar in cell-free fat-free mammary secretions from LPS-infused and control glands. These data indicate that intramammary infusion of LPS at cessation of milking did not alter growth of gram-positive mastitis pathogens in mammary secretion during the nonlactating period. Stage of lactation and the nonlactating period influenced bacterial growth and marked differences between bacteria and among strains of a bacterial species were observed. Staphylococcus aureus grew well in secretions collected during late lactation, but growth decreased during early- and mid-involution and increased again in secretions obtained near parturition. Streptococcus agalactiae and Strep. uberis grew better in mammary secretion obtained during involution than in secretions collected during late or early lactation. Streptococcus dysgalactiae grew well in mammary secretions at all time periods. These data demonstrate the variability of mastitis pathogen growth during physiologic transitions of the bovine udder.  相似文献   

3.
参照GenBank发表的序列，在金黄色葡萄球菌、无乳链球菌和停乳链球菌16SrRNA与23SrRNA之间的区域设计了3对引物，参照念珠菌和隐球菌的18SrRNA的序列设计1对引物，建立了检测金黄色葡萄球菌、无乳链球菌、停乳链球菌和酵母真菌4种乳腺炎主要致病菌的多重PCR方法。参照Skladny的方法制备模拟了细菌感染l临床标本。结果表明：本试验建立的多重PCR方法具有较好的特异性，多重PCR方法检测乳样中的金黄色葡萄球菌的细菌最小浓度为10^4CFU／mL，检测无乳链球菌、停乳链球菌和酵母真菌的细菌最小浓度分别为10^4CFU／mL、10^3CFU／mL和10^3CFU／mL。通过对采自临床型乳腺炎（46个）和隐性乳腺炎（167个）动物共计213个乳样分别用传统细菌学培养法和多重PCR方法进行检测，多重PCR对金黄色葡萄球菌和酵母真菌的检测具有更高的检出率（P〈0．01），但该方法对无乳链球菌和停乳链球菌的检出率与培养法差异不显著（P〉0．05）。  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.
Time-related concentrations in milk of a combination of trimethoprim-sulphadiazine (TMP-SDZ) intramammary formulated infusion and its relationship with pathogenic bacteria strains minimum inhibitory concentrations (MICs) isolated from clinical mastitis cows were analysed. The MICs study was performed for Escherichia coli, Staphylococcus aureus and Streptococcus sp. strains. The SDZ concentrations in milk were analysed using high-performance liquid chromatography (HPLC) and TMP using a microbiological assay. Ten lactating cows milked three times daily were used in the time-concentration studies of TMP-SDZ. Milk samples (approximately 20 mL) from the treated mammary quarters were taken at 6, 12, 24, 30 and 36 h after first administration. In order to define the withdrawal time, milk samples from the treated mammary quarters were taken at 24, 36, 48, 72, 84 and 96 h, after finishing the therapy. The MICs fluctuated between 1 and 8 microg/mL. Effective therapeutic concentrations lasted for 36 h when intramammary infusion was repeated three times every 12 h. No TMP was detected in milk for 24 h after finishing therapy. Milk SDZ concentrations were below 0.1 microg/mL in all treated cows after 84 h finishing therapy. At 96 h after finishing therapy, no SDZ milk concentrations were found in six animals, although four animals of the experimental group still had concentrations of 0.07 microg/mL.  相似文献   

6.
从宁夏9个县(区)的16个规模化牛场采集327份患乳腺炎奶牛乳样,通过分离培养和生化试验,检出含细菌乳样226份,乳样细菌检出率69.11%(226/327);鉴定出奶牛乳腺炎病原菌5大类9种260株,确认宁夏部分地区奶牛乳腺炎主要病原菌为凝固酶阴性葡萄球菌、停乳链球菌、金黄色葡萄球菌、大肠埃希菌和酵母菌。乳腺炎的发生以单一病原感染为主,占84.96%(192/226);混合感染较少,占15.04%(34/226)。  相似文献   

7.
A 31-year record-based retrospective study was carried out to determine the aetiology and temporal distribution of bovine clinical mastitis in smallholder dairy herds in the Dar es Salaam region of Tanzania over the period November 1971-December 2002. Laboratory information on 1964 quarter samples from 1365 cows in 281 smallholder dairy herds were retrieved, compiled and studied. Eighty-eight percent of the quarter samples were culture-positive and the predominant mastitis pathogens isolated were Staphylococcus aureus (25.7%), Streptococcus agalactiae (15.4%), Klebsiella pneumoniae (14.3%) and Escherichia coli (14.1%). Other isolates included Pseudomonas aeruginosa (7.5%), Streptococcus dysgalactiae (5.2%) and Streptococcus uberis (4.2%). Contagious mastitis pathogens were isolated from 45.6% of the culture-positive samples, whereas environmental and miscellaneous pathogens were isolated from 48.2% and 5.7%, respectively. Thirty percent of the miscellaneous mastitis pathogens were Candida species. The results demonstrate a steady increase in clinical Candida albicans mastitis. The prevalence of Candida albicans has increased from 1% in 1971 to 17.0% in November 2002. Conversely, despite some fluctuations, the prevalence of Staphylococcus aureus, Streptococcus agalactiae, E. coli and K. pneumoniae remain above 10%. The possible risk factors for these observations are discussed.  相似文献   

8.
Antibacterial Properties of a Silver Chloride-Coated Nylon Wound Dressing   总被引：2，自引：0，他引：2  
OBJECTIVE: A silver chloride-coated nylon wound dressing (Ag-WD) was evaluated in vitro for antimicrobial activity against five common equine wound pathogens. STUDY DESIGN: Bacterial susceptibility study. SAMPLE POPULATION: Equine wound pathogens: Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus equi subspecies zooepidemicus, and Staphylococcus aureus. METHODS: An inoculum of each pathogen was incubated directly with Ag-WD and quantitated after 24 to 48 hours of incubation. To determine if bactericidal activity of Ag-WD was contact dependent, an inoculum of E. coli and Staphylococcus aureus was incubated separately from Ag-WD by a filter and quantitated after 18 hours of incubation. Inductively coupled plasma emission spectrometry (ICP) determined the silver concentration of Mueller-Hinton broth containing Ag-WD after 24 hours of incubation. To establish if the rate of bacterial killing by Ag-WD differed from a constant silver concentration, pathogens were exposed to a silver concentration of 6.45 microg/mL and quantitated after 18 hours. RESULTS: Direct exposure to Ag-WD significantly reduced bacterial numbers after 15 minutes for K. pneumoniae, 30 minutes for E. coli, 1 hour for P. aeruginosa, and 2 hours for S. equi subspecies zooepidemicus and Staphylococcus aureus. Indirect exposure to Ag-WD resulted in > or =99.9% and > or =90% kill of the inoculum doses of E. coli at 2 hours and Staphylococcus aureus at 18 hours, respectively. Incubation of the pathogens at the constant silver concentration resulted in bacterial killing rates similar to those obtained by incubation with Ag-WD. CONCLUSIONS: In vitro, equine pathogens are effectively killed when exposed to Ag-WD, and the rate of bacterial killing by Ag-WD is similar to a constant silver concentration of 6.45 microg/mL. CLINICAL RELEVANCE: The in vitro antimicrobial properties of this silver-coated nylon wound dressing are promising for future prevention of equine wound infections.  相似文献   

9.
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   

10.
Research on mastitis in dairy cows caused by Escherichia coli has reported the emergence of strains capable of inducing chronic mastitis and that these strains adhered to and internalized into bovine mammary epithelial cells better than strains of E. coli isolated from acute mastitis. To understand mechanisms and strategies used by chronic E. coli strains to survive intracellularly internalization studies using bovine mammary epithelial cells treated with inhibitors of caveolae-mediated endocytosis (CME) and receptor-mediated endocytosis (RME), double immunofluorescence labeling confocal laser and fluorescence microscopy were conducted. Internalization studies showed that strains chronic E. coli strains persisted intracellularly longer than acute E. coli strains. Treatment of bovine mammary epithelial cells CME or RME inhibitors resulted in lower numbers of intracellular E. coli strains associated with chronic or acute mastitis than untreated controls. In addition, when selective CME inhibitors were used significantly fewer chronic E. coli were detected intracellularly than acute E. coli or untreated controls. Confocal laser microscopy showed that chronic E. coli strains colocalized preferentially with caveolae whereas acute strains did so with early endosomes, an early step of RME. These results suggest that strains of E. coli associated with chronic mastitis exploit lipid rafts/CME to internalize into and move through mammary epithelial cells. By exploiting this endocytosis pathway, chronic E. coli strains avoid bactericidal mechanisms such as endosome acidification and endosome-lysosome fusion, thus allowing intracellular survival. Data from this study helps to explain how these strains are capable of causing chronic E. coli mastitis.  相似文献   

11.
The purpose of this study was to investigate the interaction between Escherichia coli and primary mammary epithelial cell cultures derived from cows with persistent intramammary infection (IMI). Two strains of E. coli, isolated from the milk of two different cows suffering from persistent E. coli IMI were tested for adhesion to and invasion of three primary mammary epithelial cell cultures derived from mammary biopsies of the two infected cows. Intracellular E. coli were detected during five days post infection in vitro. Both strains of E. coli adhered to and invaded monolayers of all three primary mammary epithelial cell cultures. One strain adhered less but invaded more than the other. Comparison with other mammary pathogens indicated that E. coli invaded the cells less efficiently than Staphylococcus aureus, about as efficiently as Streptococcus dysgalactiae and more efficiently than Streptococcus uberis. The mechanism of E. coli invasion was studied using the cytoskeleton disrupting agents colchicine and cytochalasin D. These compounds inhibited the invasion of E. coli. Invasion of E. coli could also be inhibited by the phosphokinase inhibitors genistein and staurosporin in a dose-dependent fashion. Phorbol-myristyl-acetate (PMA) had no effect on the invasion of E. coli. Histology of mammary tissue revealed chronic inflammatory changes in quarters that were persistently infected by E. coli. Intracellular bacteria were not detected in mammary tissue sections. Polymerase chain reaction (PCR) analysis suggested that the two strains of E. coli lacked genes encoding for bundle-forming pili (bfpA), intimin (eae) and translocated intimin receptor (tir), which are characteristic for enteropathogenic E. coli (EPEC).  相似文献   

12.
Streptococcus uberis is the most common environmental mastitis pathogen causing udder inflammations of different severities in dairy cows. The aim of the study was to investigate if the different clinical outcome of mastitis induced by different strains of S. uberis can be reflected in the mammary immune response. Mammary epithelial cells and somatic milk cells were treated with heat inactivated and living S. uberis of strain A and strain B in vitro. Strain A was repeatedly isolated from a chronically infected quarter during 8 months, and persisted in the quarter despite antibiotic treatment. Strain B caused an acute clinical mastitis and was not further isolated after a single antibiotic treatment. Treatment with Strain B induced a more pronounced increase of mRNA-expression of various immune factors (interleukin-8, interleukin-1beta, RANTES, and lactoferrin) in mammary epithelial cells than strain A. In contrast to mammary epithelial cells the response of removed somatic milk cells showed no differences between the stimulation with two S. uberis strains. Tumor necrosis factor-alpha mRNA expression was not differently induced by the two strains. In conclusion, the characteristics of different severities of mastitis that are induced by different S. uberis strains in vivo can also be reflected at the level of the immune response of the mammary gland in vitro.  相似文献   

13.
The objectives of this study were to determine the serum and pulmonary disposition of tilmicosin in foals and to investigate the in vitro activity of the drug against Rhodococcus equi and other common bacterial pathogens of horses. A single dose of a new fatty acid salt formulation of tilmicosin (10 mg/kg of body weight) was administered to seven healthy 5- to 8-week-old foals by the intramuscular route. Concentrations of tilmicosin were measured in serum, lung tissue, pulmonary epithelial lining fluid (PELF), bronchoalveolar lavage (BAL) cells, and blood neutrophils. Mean peak tilmicosin concentrations were significantly different between sampling sites with highest concentrations measured in blood neutrophils (66.01+/-15.97 microg/mL) followed by BAL cells (20.1+/-5.1 microg/mL), PELF (2.91+/-1.15 microg/mL), lung tissue (1.90+/-0.65 microg/mL), and serum (0.19+/-0.09 microg/mL). Harmonic mean terminal half-life in lung tissue (193.3 h) was significantly longer than that of PELF (73.3 h), bronchoalveolar cells (62.2 h), neutrophils (47.9 h), and serum (18.4 h). The MIC90 of 56 R. equi isolates was 32 microg/mL. Tilmicosin was active in vitro against most streptococci, Staphylococcus spp., Actinobacillus spp., and Pasteurella spp. The drug was not active against Enterococcus spp., Pseudomonas spp., and Enterobacteriaceae.  相似文献   

14.
The in vitro susceptibility to penicillin G, erythromycin and clindamycin was determined by the disc diffusion test and by E-test for a total of 47 streptococcal strains (three Streptococcus uberis, 36 Streptococcus agalactiae, eight Streptococcus dysgalactiae spp. dysgalactiae) isolated from bovine intramammary infections in Argentina. Moreover, resistance phenotypes of erythromycin-resistant streptococcal isolates was characterized. MIC90 of penicillin G, erythromycin and clindamycin for S. agalactiae were 0.75, 8.0 and 12.0 microg/ml respectively. Resistance to erythromycin and clindamycin was detected in 13 (27.6%) and 12 (25.5%) isolates respectively. No isolate was resistant to penicillin G. Resistance against macrolides, lincosamides and streptogramin B (MLS(B)) represented by the constitutive MLS(B) phenotype was present in 11 (23.4%) erythromycin-resistant isolates and two isolates (4.3%) expressed the M phenotype. The inducible MLS(B) phenotype was not identified. Results suggest that beta-lactams are the first-line antibiotics when treating streptococcal udder infections; however, the continuous monitoring of the antibiotic resistance is essential, as the emergence of resistant strains has become a growing concern on the therapy of bovine mastitis.  相似文献   

15.
本研究旨在调查天津市原料奶细菌总数、体细胞数及乳房炎病原菌、耐药基因,了解全市原料奶的质量状况及引起奶牛乳房炎发生的主要原因。采集天津市5家乳品加工企业奶罐车的原料奶样品,用于检测体细胞数和菌落总数;采集天津市奶牛养殖场储奶罐奶样品,用于检测体细胞数;采集奶牛场临床型乳房炎发病乳区的牛奶样品,用于检测乳房炎病原菌及耐药基因。结果显示:2018年天津市乳品加工企业原料奶体细胞数平均值为44.65万/mL,标准差42.41万/mL,变异系数94.97%,最大值为225.50万/mL,最小值为1.20万/mL,SCC≤50万/mL的样品占74.37%,50万/mL200万/mL的样品占3.13%;细菌总数平均值11.54万CFU/mL,标准差26.28万CFU/mL,变异系数227.66%,最大值190.00万CFU/mL,最小值0.095万CFU/mL,细菌总数≤10万CFU/mL的样品占74.37%,10万CFU<细菌总数≤50万CFU/mL的样品占21.25%,50万CFU<细菌总数≤100万CFU/mL的样品占1.88%,100万CFU/mL<细菌总数≤200万CFU/mL的样品占2.50%。2018年天津市奶牛养殖场原料奶体细胞数平均值为38.81万/mL,标准差36.49万/mL,变异系数94.03%,最大值为210.00万/mL,最小值为0.80万/mL,SCC≤50万/mL的样品占79.17%,50万/mL200万/mL的样品占0.83%。乳房炎病原菌检测结果显示:36个样品检出病原菌,总检出率为94.74%;共检出8种病原菌,检出率最高的是乳房链球菌,检出率为73.68%,其他病原体检出率依次为:牛支原体34.21%,牛棒状杆菌13.16%,无乳链球菌10.53%,大肠杆菌5.26%,白色念球菌5.26%,停乳链球菌2.63%,铜绿假单胞菌2.63%。14个样品检出耐药基因,总检出率为36.84%;2种耐药基因的检出率分别为β-内酰胺耐药基因CTX-M934.21%,耐甲氧西林葡萄球菌耐药基因MecA 2.63%。研究表明,2018年天津市原料奶SCC及细菌总数大部分接近欧盟标准,但仍有待进一步提高;引起天津地区临床型乳房炎的3种主要致病菌为乳房链球菌、牛支原体和牛棒状杆菌。  相似文献   

16.
The binding of bovine complement S protein (vitronectin) to Streptococcus dysgalactiae isolates from cattle with mastitis and the S protein's role in streptococcal adherence to bovine epithelial cells were investigated. All 25 clinical isolates of S dysgalactiae interacted with bovine S protein. None of the other streptococcal species tested bound to bovine S protein. The S protein-binding sites were saturable and highly sensitive to trypsin. The binding of bovine S protein to S dysgalactiae isolates was specific and could not be inhibited by other plasma proteins, such as fibronectin, albumin, fibrinogen, alpha 2-macroglobulin, or IgG. Similarly, streptococcal binding of bovine S protein was not influenced by the synthetic peptide Gly-Arg-Gly-Asp-Ser, which constituted the host cell attachment sequence of S protein. In adherence experiments, prior binding of bovine S protein to S dysgalactiae enhanced streptococcal adherence to bovine epithelial cells. The enhancing effects by bovine S protein were abolished when the respective binding sites on the streptococci were digested by trypsin. Thus, bovine S protein could be an important mediator of adherence of S dysgalactiae to bovine epithelial cells.  相似文献   

17.
A monitoring programme conducted in Europe since 1994 to survey the marbofloxacin susceptibility of bacterial pathogens isolated from cattle has established the susceptibility of bacterial strains isolated before any antibiotic treatment from bovine mastitis and bovine respiratory disease (BRD) cases between 2002 and 2008. Minimum inhibitory concentration (MIC) was determined by a standardised microdilution technique. For respiratory pathogens, Pasteurella multocida and Mannheimia haemolytica isolates (751 and 514 strains, respectively) were highly susceptible to marbofloxacin (MIC≤0.03 μg/ml for 77.39 per cent of the strains) and only 1.75 per cent of M haemolytica strains were resistant (MIC≥4 μg/ml). Histophilus somni isolates (73 strains) were highly susceptible to marbofloxacin (0.008 to 0.06 μg/ml). Mycoplasma bovis MIC (171 strains) ranged from 0.5 to 4 μg/ml. For mastitis pathogens, the majority of Escherichia coli isolates were highly susceptible to marbofloxacin (95.8 per cent of 617 strains). Staphylococcus aureus and coagulase-negative staphylococci (568 and 280 strains) had a homogenous population with MIC centred on 0.25 μg/ml. Streptococcus uberis and Streptococcus dysgalactiae (660 and 217 strains) were moderately susceptible with MIC centred on 1 μg/ml. Marbofloxacin MIC for these various pathogens appeared stable over the seven years of the monitoring programme and was similar to previously published MIC results.  相似文献   

18.
连翘酯苷在体外与体内的抑菌效果研究   总被引：7，自引：0，他引：7  
本文提取制备了连翘酯苷,并对其进行了体内和体外的抑菌试验。试验结果表明,连翘酯苷在体外和体内对引发奶牛乳房炎的主要致病菌金黄色葡萄球菌、停乳链球菌、无乳链球菌均有较好的抑制作用。  相似文献   

19.
为筛选和研究可有效抑制奶牛乳腺炎主要致病菌的中药复方及其抗炎效果,本文通过对黄连、黄柏、赤芍、苦参、连翘等5味中药进行正交试验设计组方,采用平板打孔法、试管二倍稀释法分别研究了各单味中药及其复方对金黄色葡萄球菌、停乳链球菌、无乳链球菌和大肠杆菌的体外抑菌效果,同时研究了筛选出的中药复方的不同剂型对二甲苯致小鼠耳肿胀的急性炎症和棉球致肉芽肿的慢性炎症的抑制效果。结果表明,4株菌对5味中药的敏感性强弱依次为无乳链球菌、停乳链球菌、金黄色葡萄球菌、大肠杆菌;中药方3和方7对4种菌的体外抑菌活性较强,且以方3的搽剂及膏剂对急、慢性炎症具有良好的抑制效果。本研究结果为研制防治临床奶牛乳腺炎的药物提供了试验依据。  相似文献   

20.
本试验应用组织块培养和差速消化法获取原代奶牛乳腺上皮细胞,免疫荧光鉴定正确后MTT法评价不同浓度的胰岛素样生长因子-Ⅰ（insulin-like growth factor-Ⅰ,IGF-Ⅰ）、肝细胞生长因子（hepatocyte growth factor, HGF）、转化生长因子-β1（transforming growth factor-β1,TGF-β1）、干扰素-γ （interferon-γ,IFN-γ）对奶牛乳腺上皮细胞体外增殖的影响。结果表明,IGF-Ⅰ、HGF分别在 10～200和0.1～100 ng/mL对乳腺上皮细胞的增殖呈正相关,而TGF-β1（2.5～100 ng/mL）、IFN-γ（5～160 ng/mL）则剂量依赖性地抑制乳腺上皮细胞增殖。提示,IGF-Ⅰ、HGF均能促进奶牛乳腺上皮细胞的体外增殖,该作用在一定浓度范围内有剂量依赖性;TGF-β1、IFN-γ对乳腺上皮细胞的增殖作用出现剂量抑制效应。  相似文献   

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