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1.
Mastitis is inflammation of mammary gland affecting all the species of domestic animals. Fragments of the mitochondrial genome released from dying cells are considered surrogate markers of mitochondrial injury. We hypothesized that bovine mastitis would be associated with increased cell free mitochondrial DNA (mtDNA) content in serum and milk. Milk and serum samples were collected from sub-clinical mastitic and normal animals. Mastitis was confirmed by California mastitis test and bacterial isolation. Oxidative stress, nitric oxide and inflammatory cytokines were also estimated. Real time polymerase chain reaction was conducted in serum and milk from sub-clinical mastitic animals and compared with healthy animals targeting the mtDNA genes cytochrome b. Mastitis animals showed higher oxidative stress markers and nitric oxide along with higher level of inflammatory cytokines. Cell free mtDNA was significantly higher in serum and milk of mastitic animals comparing to that of healthy control. The higher cell free relative mtDNA content in mastitis animals indicates injury to the mammary epithelial cells and thereby releasing the mtDNA in the milk and blood. This mtDNA may be a bio-marker of oxidative stress and tissue injury in bovine mastitis.  相似文献   

2.
In vitro methods were used to test the hypothesis that Escherichia coli from bovine mastitis are essentially no different from isolates from bovine feces. Fifty E. coli isolates from bovine mastitic milk, 50 from feces of mastitic cows and 50 from feces of healthy cows were compared with respect to biochemical properties and certain potential virulence factors. There were no significant differences among the groups in tests for biotype; production of colicins, colicin V, or Vero cell cytotocity; and growth in 90% gnotobiotic calf serum or 90% normal milk whey. Resistance to killing in 90% gnotobiotic calf serum varied from 66 to 84%. Most isolates grew in normal whey: the percentage in a group varied from 86 to 96. Mastitic milk isolates were significantly different from the fecal isolates in adonitol fermentation (P0.006), production of aerobactin (P0.026), and ability to grow in 90% mastitic whey (P0.00004). However, only 40% of mastitis E. coli fermented adonitol and only 20% produced aerobactin. Ninety-six percent of mastitic milk E. coli grew in mastitic whey, whereas 64% and 60%, respectively, of mastitic fecal and normal fecal isolates grew in this medium. It is concluded that none of the properties that were investigated constitute potential virulence factors or markers for ability to induce mastitis; the data are consistent with the hypothesis that mastitic E. coli are simply opportunistic pathogens.  相似文献   

3.
This study was conducted to investigate polymorphism of the coagulase gene of Staphylococcus aureus causing bovine mastitis. One hundred eighty-seven strains of S. aureus were isolated from bovine mastitic milk samples obtained from 187 different Danish dairy farms. The isolates were characterised for restriction fragment length polymorphism (RFLP) of the coagulase gene. A variable region of the coagulase gene was amplified using the polymerase chain reaction (PCR) followed by AluI restriction enzyme digestion. A total of 15 different RFLP patterns were observed. The predominant pattern was found in 35% of the isolates. The ease of analysing coagulase gene polymorphisms among a large number of strains, and the multiple distinct polymorphic patterns generated, supports the use of this technique in epidemiological investigations of bovine mastitis. The predominating variants may have predelection for causing intramammary infections.  相似文献   

4.
Two methods for estimating fat and cream content in cow's milk were compared in normal and mastitic milk. Fat content in milk from periparturient cows was estimated simultaneously by the method of Gerber and by simply spinning small samples of untreated milk in a haematocrit centrifuge. Mastitis was experimentally induced by inoculating mammary glands with Escherichia coli. On the average, cream content in milk from normal, and uninfected mammary glands measured by the haematocrit method was 20 to 23% higher than milk fat measured by the method of Gerber. In mastitic milk from infected mammary glands haematocrit readings were 40% higher than those of Gerber. Milk fat values as measured by both methods were highly correlated. It is concluded that the haematocrit method can be used for rapid estimation of milk fat content in normal milk of cows. The greater difference between cream and fat content in mastitic milk compared to normal milk can probably, at least partly, be ascribed to the presence of large clots of cells and fat in the milk during mastitis. During centrifugation of the viscous mastitic milk samples, milk cells and fat probably are not completely separated. This results in overestimated readings of milk cream content with the haematocrit method. It therefore seems advisable to use the Gerber method for accurate measurements of milk fat in mastitic milk.  相似文献   

5.
在对山东7个地区14个奶牛场临床型和隐性乳腺炎调查的基础上采集234头临床乳腺炎病牛乳样、241个隐性乳腺炎乳样并分别做了细菌学检查,结果表明:泌乳期临床型乳腺炎病原菌以凝固酶阴性葡萄球菌、金黄色葡萄球菌、链球菌、酵母菌和棒状杆菌为主;干奶期临床型乳腺炎病原菌以大肠杆菌、链球菌、金黄色葡萄球菌、凝固酶阴性葡萄球菌和酵母菌为主;隐性乳腺炎病原菌以凝固酶阴性葡萄球菌、金黄色葡萄球菌链球菌、酵母菌、假单胞菌和棒状杆菌为主;厌氧菌在隐性乳腺炎、干奶期乳腺炎和干奶期乳腺炎乳样的捡出率分别为 5．82％,4．17%,10．16％;隐性乳腺炎、干奶期乳腺炎细菌的共感染率较高,与泌乳期乳腺炎病原菌的差异极显著(P<0．01),隐性乳腺炎与干奶期乳腺炎病原菌共感染率差异不显著(P >0．05)。  相似文献   

6.
甘肃地区牛源金黄色葡萄球菌分子鉴定及RAPD分型   总被引：1，自引：0，他引：1  
本研究目的是分离鉴定引起甘肃地区奶牛乳房炎的金黄色葡萄球菌,掌握其基因型情况。利用16S、23SrRNA保守序列PCR扩增对乳房炎奶样中的金黄色葡萄球菌进行鉴定,并进行RAPD基因分型。结果表明,310份奶样中共分离出金黄色葡萄球菌100株,RAPD结果显示这100株金黄色葡萄球菌均可得到清晰的RAPD指纹图谱,扩增产物在2～7条带之间,具有多种带型组成。通过聚类分析100株菌产生11个基因型,其中Ⅰ型4株,Ⅱ型4株,Ⅲ型10株,Ⅳ型13株,Ⅴ型7株,Ⅵ型24株,Ⅶ型16株,Ⅷ型6株,Ⅸ型4株,Ⅹ型10株,Ⅺ型2株。Ⅵ型为该地区的流行优势菌群,不同牛场各基因型菌株分布有明显差异。本研究说明牛场的环境与养殖条件对病原菌流行传播有明显的影响,这一结果对地区性奶牛乳房炎的防治提供了可靠的理论依据。  相似文献   

7.
奶牛隐性乳房炎的流行病学调查及病原菌的分离鉴定   总被引：5，自引：0，他引：5  
对某大型奶牛场进行隐性乳房炎的流行病学调查,以及致病菌的分离鉴定,结果得出:对同一牛舍76头泌乳牛进行CMT检测,其中隐性乳房炎的牛发病率为68.42%(52/76),在278个有效乳区中,隐性乳房炎的乳区有109个,乳区阳性率为39.21%(109/278),从10份被检乳汁中共分离出59株细菌,经生化鉴定其主要致病菌为金黄色葡萄球菌、无乳链球菌和大肠杆菌,分别占总数的18.64%。  相似文献   

8.
A within cow comparison was made between milk progesterone levels in healthy and mastitic quarters. Material was collected from cows with mastitis induced by bacterial inoculation, or by inoculation with bacterial endotoxin. Furthermore material from cows with spontaneous subclinical mastitis was used. Milk progesterone levels were lowered due to mastitis. However, the decrease was not large enough to cause misinterpretation of where in the oestrous cycle (luteal phase or non-luteal phase) the samples were taken.  相似文献   

9.
To determine the effects of ozone on the phagocytosis of bovine polymorphonuclear leukocytes (PMNs), ozone gas was administered in vitro on the blood and milk of healthy lactating cows, cows with acute mastitis, and cows with milk fever. In the blood of healthy dairy cattle, although there was no significant effect of ozone gas on the viability of the leukocytes, phagocytosis of PMNs significantly decreased. In contrast, ozone gas administration in vitro significantly increased phagocytosis of PMNs from the blood of cows with acute mastitis and milk fever, and from mastitic milk. These findings showed that ozone administration in vitro has positive and negative effects on bovine PMN phagocytosis, depending on the health status of the animal.  相似文献   

10.
One hundred and sixty-six Staphylococcus aureus isolates from mastitic milk samples from different cows on 26 farms were investigated for staphylococcal enterotoxins(SEs) and toxic shock syndrome toxin-1(TSST-1) by polymerase chain reaction(PCR) and reverse passive latex agglutination assay(RPLA). SEs and the TSST-1 gene were detected in thirty-seven isolates based on a multiplex PCR; SEA was detected in 32 isolates, SEB in 3 isolates, SEC in 1 isolate, and SEA and the TSST-1 gene in 1 isolate. Of the 37 enterotoxigenic isolates, thirty-three isolates were enterotoxigenic according to RPLA, where 29 isolates produced SEA, 3 isolates produced SEB, and 1 isolate produced SEC. The enterotoxin-producing S. aureus isolates were further characterized by pulsed-field gel electrophoresis(PFGE). A macrorestriction analysis revealed 11 PFGE patterns. Among the 33 enterotoxigenic S. aureus isolates, 45.4% exhibited the same PFGE pattern I. Accordingly, although the enterotoxin-producing S. aureus isolates from bovine mastitis were genetically diverse, 1 common genotype prevailed on the farms, indicating that PFGE pattern I isolates may be the most disseminated in Korea.  相似文献   

11.
The concentration of the nuclease produced by Staphylococcus aureus and titres of antinucleases were determined in the udder secretions of 539 cows, most of them with clinical S. aureus mastitis. On average, the nuclease was detected in 85 % of the samples from mastitic quarters. In milk from most cows, antinucleases neutralize the activity of the nuclease, and a successful demonstration of this thermostable enzyme presupposed the inactivation of antibodies by heating the samples on boiling water-bath. The nuclease was demonstrated most frequently, and at highest concentration, in samples from cows with a severe mastitis and from cows which did not recover completely.The test for nuclease, performed out both non-heated and heated quarter samples, and the antimuclease test, may supplement conventional laboratory methods. The advantages and limitations of the methods in the diagnosis of mastitis are discussed.  相似文献   

12.
选取临床型乳房炎自然发病病例9头,乳房内灌注乳酸链球菌素(Nisin)进行治疗,通过检测日产奶量以及停药后2,7,14,21 d的主要乳成分指标,确定Nisin乳房灌注剂对临床型乳房炎病例泌乳性能的影响情况。结果显示,乳房内灌注Nisin后患病乳区乳腺组织得到一定程度的修复,日产奶量逐渐恢复,停药后8 d的日产奶量与发病前6 d相比,平均降幅2.5 kg。治愈后患病乳区牛奶乳脂肪、乳蛋白、乳糖和非脂乳固体含量都呈现上升趋势,但与同期相比,略低于非用药乳区混合牛奶;其中,乳糖含量增加幅度较快,除停药后2 d的治疗乳区外,乳糖含量均在4.7%以上,提示奶牛乳房炎病例经Nisin乳房灌注治疗后,受损乳腺组织修复较快,乳腺上皮细胞合成乳成分的能力大幅度提高。由此可见,Nisin对奶牛临床型乳房炎具有良好的治疗作用。  相似文献   

13.
New tools are needed to detect chronic sub-clinical mastitis, especially in automatic milking systems. Haptoglobin and serum amyloid A (SAA) are the two most sensitive bovine acute phase proteins, and their concentrations increase in milk from cows with clinical mastitis and in milk from cows with experimentally induced chronic sub-clinical Staphylococcus aureus mastitis. The aim of this study was to further evaluate the potential for haptoglobin and SAA in milk as indicators of chronic sub-clinical mastitis. Quarter milk samples were collected from 41 cows with a mean composite milk somatic cell count (CSCC) above 300,000 cells/mL during at least two months prior to sampling. Quarter milk samples were also taken from eleven cows with a mean CSCC below 80,000 cells/mL during at least two previous months. These samples were analysed for haptoglobin, SAA, adenosine triphosphate (ATP) activity and bacterial growth. The samples were grouped according to their ATP, haptoglobin and SAA status. ATP+ samples had ATP > 2 x 10(-10) mol/mL, Hp+ and SAA+ samples had detectable levels of haptoglobin (> or = 0.3 mg/L) and SAA (> or = 0.9 mg/L), respectively. In udder quarter samples from healthy cows, 42 out of 44 samples belonged to the ATP-Hp-SAA- group. Among cows with chronic sub-clinical mastitis, the ATP+Hp+SAA+ group contained 66 out of 164 samples while 44 samples belonged to the ATP+Hp-SAA- group. Detectable levels of haptoglobin and SAA were found in 92 and 80 samples, respectively. Growth of udder pathogens was detected in 28 samples and Staphylococcus aureus was the most common bacteria. In conclusion, haptoglobin and SAA concentrations below the detection limit were considered as good indicators of healthy udder quarters. A substantial variation in haptoglobin and SAA concentrations in milk was observed in udder quarters with chronic sub-clinical mastitis.  相似文献   

14.
To determine the prevalence of capsular polysaccharide (CP) types of Staphylococcus aureus isolated from bovine mastitic milk in Korea, the protective effect of the conjugates, composed of microencapsulated S. aureus clinical isolate type 8 CP bound to Pseudomonas aeruginosa exotoxin A (ETA) was evaluated in mice. Of 107 S. aureus isolates, serotype 5 and 8 accounted for only 26 or 24.2%. When serotype 336 antiserum was employed, fifty of the remaining 81 isolates were typed as 336, 26 reacted with two serotypes, and 5 were nontypeable. Mice challenged with the same strain used for immunization had fewer S. aureus cells in their kidneys than mice challenged with the heterologous strain. But the magnitudes of difference on bacterial clearance were similar in both groups, indicating that the significance of this result remains to be determined. Mice immunized with the conjugate elicited an antibody response 3 days post injection, which persisted for 13 days of the observation period after second injection in some mice. The mice immunized with the CP8-ETA conjugates developed antibodies significantly higher than those immunized with CP-Freund's adjuvant or PBS. In in vivo bacterial challenge experiment, the survival rate of mice immunized with CPS-ETA conjugate was significantly higher than that of mice immunized with PBS. It was suggested that CP8-ETA vaccine had a potential to protect mice against experimental S. aureus bacteremia.  相似文献   

15.
Eleven Escherichia coli isolates from clinical bovine mastitis cases (mastitic strains) and 11 from the cowshed environment (environmental strains) were compared, to determine if the former were a subset of the latter. The mastitic and environmental strains could not be distinguished according to O antigen and antibiotic sensitivity. All mastitic isolates showed significantly (P<0.0001) faster growth in milk and faster lactose fermentation than most (approximately 64%) environmental strains, but growth rates in nutrient broth did not differ. The rates of lactose fermentation and growth in milk were positively correlated. Adhesion and phagocytosis of mastitic strains by bovine PMN were significantly (P<0.0001) lower than those of environmental strains, and correlated negatively with growth in milk and lactose fermentation. The average percentages of killing by bovine leukocytes in the two sources were not statistically different. All mastitic strains were serum sensitive, whereas most ( approximately 72%) environmental ones were resistant. Finally, pulse-field gel electrophoresis revealed two main pulse type clusters, sharing a similarity coefficient of 79%. Cluster 1 comprised only environmental strains, whereas cluster 2 comprised mostly mastitic strains and only three environmental ones. Four mastitic strains shared a similarity coefficient of less than 74% with the other strains and were not included in the clusters. Our results suggest that clinical bovine mastitis E. coli isolates may form a subset of the general environmental E. coli population; they seem better able to multiply in the udder medium and to evade the host cellular innate immune response, and are genetically distinct from most environmental strains.  相似文献   

16.
Lactoferrin concentration (LFC) in normal and mastitic milk of dairy goats were examined. LFC in bulk milk collected from 70 dairy goat farms and individual milk samples from 10 goats with mastitis were measured by enzyme-linked immunosorbent assay (ELISA) and their reaction time in methylene blue reduction test (MBRT) monitored. Bulk milk samples were categorized into three grades, such as high, normal and low, based on the reaction time in MBRT. The mean LFC in milk that was considered high quality (167 microg/ml) was significantly lower than that of those graded as normal (218 microg/ml) and low quality (304 microg/ml), while mean LFC in mastitic milk was 587 microg/m l. The correlation coefficient between milk LFC and MBRT time was found to be -0.7. Three goats were inoculated with Staphylococcus aureus into one of their udder halves. The mean milk LFC was found to be significantly higher (1500 microg/ml) than the control (30 microg/ml). These findings suggest that milk LFC may be useful as an index for intramammary infection in goats.  相似文献   

17.
Combined parenteral and intramammary treatment of mastitis caused by Staphylococcus aureus was compared to parenteral treatment only. Cows with clinical mastitis (166 mastitic quarters) caused by S. aureus treated by veterinarians of the Ambulatory Clinic of the Faculty of Veterinary Medicine during routine farm calls were included. Treatment was based on in vitro susceptibility testing of the bacterial isolate. Procaine penicillin G (86 cases due to beta-lactamase negative strains) or amoxycillin-clavulanic acid (24 cases due to beta-lactamase positive strains) was administered parenterally and intramammarily for 5 days. Efficacy of treatments was assessed 2 and 4 weeks later by physical examination, bacteriological culture, determination of CMT, somatic cell count and NAGase activity in milk. Quarters with growth of S. aureus in at least one post-treatment sample were classified as non-cured. As controls we used 41 clinical mastitis cases caused by penicillin-susceptible S. aureus isolates treated with procaine penicillin G parenterally for 5 days and 15 cases due to penicillin-resistant isolates treated with spiramycin parenterally for 5 days from the same practice area. Bacteriological cure rate after the combination treatment was 75.6% for quarters infected with penicillin-susceptible S. aureus isolates, and 29.2% for quarters infected with penicillin-resistant isolates. Cure rate for quarters treated only parenterally with procaine penicillin G was 56.1% and that for quarters treated with spiramycin 33.3%. The difference in cure rates between mastitis due to penicillin-susceptible and penicillin-resistant S. aureus was highly significant. Combined treatment was superior over systemic treatment only in the beta-lactamase negative group.  相似文献   

18.
Strains of Staphylococcus species isolated from bovine mastitic milk at 66 dairy farms in Japan during the period from November 1988 to May 1989 were identified, and examined for their drug susceptibility and beta-lactamase production in order to clarify an epidemiological aspect of bovine mastitis caused by staphylococci. The results of bacteriological identification showed that the most predominant species was S. xylosus. Other major species isolated were S. aureus, S. sciuri and S. hyicus. Thirty-eight (71.7%) isolates of S. xylosus, 21 (45.7%) of S. aureus and 5 (71.4%) of S. epidermidis were positive for beta-lactamase production. Most of the beta-lactamase-producers of S. aureus were classified as high producers, although all of the beta-lactamase-positive S. xylosus isolates remained to be low producers. All isolates of S. aureus were sensitive to methicillin and cloxacillin at 6.25 micrograms/ml and 1.56 micrograms/ml, respectively, and none of methicillin-resistant S. aureus were detected. Isolates of other species were considered to be susceptible to 6 beta-lactams, in contrast to human isolates, but antibacterial activities of penicillin G and ampicillin were affected more strongly by beta-lactamase than those of methicillin, cloxacillin, cefazolin and cefoperazone.  相似文献   

19.
Strains of Staphylococcus aureus were isolated from bovine mastitis, subcultured and maintained in the laboratory for up to 3 years. Encapsulation was assessed by production of a diffuse colony in serum-soft agar. Eight (4%) of 200 strains were encapsulated. Three rapid passages of the remaining 192 strains through either brain-heart infusion broth containing 30% serum or modified 110 medium retrieved the capsule in 75%, but this was rapidly lost after subculture on blood agar. The stimulation of capsule production was studied in 18 of these strains by addition of various components to the passaging medium. Heat-labile factors in serum, milk and mastitic milk enhanced capsule production while bovine serum albumin, an extract of polymorphonuclear leucocytes, NaCl and immunoglobulins had minimal effect. The results indicate that encapsulation is common in bovine staphylococci and while it is lost on subculture, may be retrieved under appropriate conditions.  相似文献   

20.
A total of 59 mastitis staphylococcic strains were tested for growth agglutination upon supplementation of growth media with ovine and bovine milk whey and mammary secretions from dry cows. Differences were observed when comparing bacterial species or origins (ovine vs. bovine) of bacteria and whey. All of the ovine and bovine S. aureus strains tested, but only 4 among 22 other ovine mastitis staphylococcic strains, showed growth agglutination in Todd Hewitt broth (THB) supplemented with greater than or equal to 30% (v/v) ovine milk whey. None of the strains agglutinated during growth in regular THB medium. Ovine whey had an agglutination induction capacity higher than bovine whey (P less than 0.005), concerning the number of responsive ovine and bovine S. aureus strains. There were no differences between whey samples from different ewes with regard to their capacity to induce agglutination. Ovine S. aureus strains were more responsive than bovine strains of this bacterial species, concerning the number of responsive strains (P less than 0.001) to bovine whey (greater than or equal to 30% in THB), the proportion of responsive strains at low (10%) ovine whey concentration (P less than 0.001), and the strength of reaction (precipitation timing and clump size). Secretions from dry cows systematically induced agglutination in all of the bovine and ovine S. aureus strains tested.  相似文献   

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