Effects of postmilking teat treatment on the colonization of Staphylococcus aureus on chapped teat skin

Sixteen Holstein cows were used to test the effect of postmilking teat treatment on colonization and intramammary infection by Staphylococcus aureus on chapped teats. Treatments were (1) chapping the teat and using 1% I2/10% glycerin postdip solution, (2) 1% I2/10% glycerin postdip solution on nonchapped teats, (3) chapping the teat and using 10% glycerin postdip solution, (4) chapping the teat and not using a postdip solution. All mammary glands were free of S aureus teat skin colonization and intramammary infection at the start of the study. Teats selected for chapping were dipped in 1N NaOH prior to 3 applications of S aureus broth culture; cultures were applied at 12-hour intervals on all teats. Treatments were applied after each milking for 30 days and were initiated after the second broth dip. Teat skin swab specimens and milk samples were collected before treatment application. Teat skin condition was scored daily. Nonchapped teats (treatment 2) did not support skin or orifice colonization by S aureus. Treatment-1 teats healed most rapidly and supported less colonization in skin and orifice than did treatment-3 and -4 teats. Teat skin scores and skin colonization were lower for treatment-3 than treatment-4 teats. A correlation between teat skin colonization and teat skin conditions was found. Two intramammary infections were found in treatment-4 quarters and 1 in a treatment-3 quarter.(ABSTRACT TRUNCATED AT 250 WORDS)     

产肠毒素金黄色葡萄球菌显色培养基的制备

为了研究一种用于分离与鉴别产肠毒素金黄色葡萄球菌的FS显色培养基,依据产肠毒素金黄色葡萄球菌特有的耐热核酸酶和凝固酶阳性的特点,设计对产肠毒素金黄色葡萄球菌选择性培养和耐热核酸酶联合测试的FS显色培养基。应用质控菌株,对其特异性和灵敏度进行检测。结果显示:FS显色培养基对产肠毒素金黄色葡萄球菌具较好的特异性和灵敏度,特征菌落出现时间缩短至12 h。FS显色培养基对产肠毒素金黄色葡萄球菌具较好的特异性和灵敏度,具有较高的应用价值。     

Effects of quarter individual and conventional milking systems on teat condition

An on-farm experiment was performed to investigate the effects of two milking systems on teat condition. An auto-tandem milking parlor made by GEA^® with a conventional milking cluster (CON) and a single tube milking system called MultiLactor^® (MULTI) from Siliconform GmbH were tested. For both milking system, in total 73 animals were milked over a period of seven months. To assess the effect of milking on teat condition, each teat was scored according to teat-condition evaluation systems. To evaluate teat condition, the teat color, ring formation at the teat base and formation of teat hyperkeratosis were assessed. The evaluation was conducted once a month after milking. The collected data were then statistically analyzed with generalized linear mixed models. The only differences found between the two milking systems occurred with respect to their effect on teat color. MULTI showed significantly better scores compared to CON. There were no significant differences between the milking systems with regard to teat-end hyperkeratosis and ring formation at the teat base.     

沙门菌 金黄色葡萄球菌和志贺氏菌DNA提取方法的比较

根据国家食源性疾病监测网的统计资料显示,近10年来,在我国由微生物引起的食源性疾病中毒事件中,由沙门菌、金黄色葡萄球菌和志贺氏菌引起的分别占17.9%、8.9%和5.9%[1],因此对它们进行快速检测可以为有效控制其传播和预防食物中毒提供更为有效的手段和工具[2].     

Raised herd somatic cell count due to Staphylococcus aureus following the failure of an automatic teat spraying system

This study describes the failure of a single jet exit race automatic teat spray (ATS) system resulting in the spread of Staphylococcus aureus infection in a 135-cow dairy herd, which showed an increased herd somatic cell count from 91,000/ml to 554,000/ml over a nine-month period. S aureus was isolated from 34 of 46 high cell count cows. The milking procedures were modified and manual teat spraying was restarted. Bacteriology was used to identify S aureus positive high cell count cows, and first and second lactation cows were treated during lactation. If their cell counts were not reduced, these were then culled. High cell count S aureus cows in lactation three or above were culled. The three-month geometric mean cell count fell to below 150,000/ml within five months. As all replacements were home-bred, S aureus infection must have spread from within the herd itself. All other causes have been eliminated, and this spread is attributed to the failure of the ATS to carry out effective postmilking teat disinfection. The advantages and disadvantages of ATS systems are discussed, especially in relation to robotic or voluntary milking systems.     

Comparison of four test procedures to identify Staphylococcus aureus isolated from bovine intramammary infections

A comparison was made between conventional tube coagulase, macrocupsular coagulase, latex agglutination, and miniaturized biochemical test systems for identification of Staphylococcus aureus of bovine origin. A total of 303 gram-positive, catalase-positive cocci of bovine origin were tested. Agreement between each pair of 4-hour tube coagulase, macrocupsular coagulase, latex agglutination, and miniaturized biochemical test results within isolates was greater than 95.0. Seventeen (5.6%) isolates were test negative for 4-hour tube coagulase, but test positive for 24-hour tube coagulase. Thirteen (76.5%) of these isolates were identified as S hyicus, 3 were S aureus, and 1 was not identified.     

Fluorescent amplified fragment length polymorphism genotyping of human and animal Staphylococcus aureus isolates from dairy farms with manual milking

Fluorescence amplified fragment length polymorphism (fAFLP) was used to assess the genetic relatedness of 40 Staphylococcus aureus strains isolated from human and animal skin samples in seven dairy farms with manual milking. S. aureus was isolated from 11 out of 30 (36%) human skin samples and from 29 out of 100 (29%) teat skin samples from apparently healthy cows. Genomic DNA from each isolate was double-digested with EcoRI and MseI and complementary oligonucleotide adaptors were ligated to the restriction fragments. Pre-selective and selective amplification reactions were performed, the amplified fragments were separated by electrophoresis in an ABI377 sequencer and analysed using GeneScan 3.1 and Genotyper 2.5. Three single isolates (a-c), a predominant cluster with 35 isolates (d) and another cluster with two isolates (e) were identified. Both clusters d and e included human and animal isolates genetically related, because the profiles had 90-100% homology. Since no cluster was comprised uniquely of human or animal isolates and given the close genetic relatedness among human and animal samples in the farms, the present findings support the hypothesis that dairy workers can spread S. aureus through manual milking.     

Electron microscopic study of leucocytic infiltration of the mammary teat duct during infection with Staphylococcus aureus

Leucocytic response to Staphylococcus aureus infection was observed in the bovine mammary teat duct using transmission electron microscopy. Leucocytes migrating across the stratified squamous epithelium were observed in close association with areas colonised by cocci. Leucocytes gained access across the epithelium to the teat duct lumen by: passage as luminal cells desquamated; migration through degenerate cell cytoplasm; and penetration of cell junctions. The results provide evidence of marked leucocytic infiltration into ductal tissue which may participate in the cellular response to mastitis.     

Radiographic method for anatomic study of the teat canal: changes between milking periods.

Radiographs of the teat canal (papillary duct) of 14 Holstein-Friesian cows were made immediately after removal of the milking machine and at 2, 4, 6, and 8 hours. The distal, middle, and proximal diameters of the canal were dilated when the machine was removed. The distal and the middle parts of the teat canal were more dilated than the proximal part of the canal. Two hours later, the canal was constricted in the 3 areas. During the next 6 hours, the canal dilated-the greatest kilation being at the proximal paet of the teat canal. The average lenght of the teat canal remained unchanged. Apparently, microorganisms can easily pass through the teat canal when it is dilated at the end of machine milking and for up to 2 hours after milking.     

Genotyping of Staphylococcus aureus isolated from bovine mastitis.

The present study was designed to comparatively investigate 25 Staphylococcus aureus strains isolated from bovine subclinical mastitis. The S. aureus strains, obtained from six different farms at five locations in one region of Germany, were characterized by phenotypic and genotypic methods. The S. aureus could be identified and further characterized by their cultural, biochemical and hemolytic properties. To analyze the epidemiological relationship the isolates were subjected to DNA fingerprinting by macrorestriction analysis of their chromosomal DNA, by PCR amplification of the gene encoding the 16S-23S rRNA intergenic spacer, by PCR amplification of the gene encoding the IgG binding region and the X region of protein A and by amplifying, and subsequent, digestion of the gene encoding staphylococcal coagulase. The macrorestriction analysis revealed five DNA restriction patterns with DNA patterns I, III and IV occurring in three, four, and three different farms, respectively. In addition, clones with different DNA patterns could be found within one herd. The PCR products for the spacer DNA, the spa gene encoding the X region of protein A and the coa gene encoding coagulase corresponded mostly to the pattern observed by DNA fingerprinting. Amplification of the gene encoding the IgG binding region revealed sizes of 620 bp for 20 of the isolates and 280 bp for four isolates indicating, for the latter, a deletion of segments in this region. These findings show, that single, widely distributed clones seemed to be responsible for cases of bovine subclinical mastitis found in one region of Germany.     

DNA-protein immunization against the GapB and GapC proteins of a mastitis isolate of Staphylococcus aureus

One of the most economically important diseases that affect the dairy industry is bovine mastitis caused by strains of S. aureus. The development of an effective vaccine has been hampered by the antigenic diversity of the bacterium. Immunization with plasmid DNAs, encoding S. aureus antigens either as single molecule or as chimeric products containing at least two antigens, has been proposed as a novel strategy to prevent this costly disease. We continued our studies on a chimeric protein composed of the surface-located GapB and GapC proteins of S. aureus and in this work we tested the effects of DNA vaccination with plasmids encoding the individual antigens as well as the GapC/B protein with or without a boost with the recombinant proteins. The results showed that DNA vaccination alone was unable to elicit a significant humoral response and barely able to elicit a detectable cell-mediated response to the recombinant antigens. These effects were overcome by boosting with the proteins indicating that these DNA vaccines alone were not sufficient to mount an immune response against the S. aureus GapB and GapC proteins.     

Differential responses of cells from human skin keratinocyte and bovine mammary epithelium to attack by pore-forming Staphylococcus aureus alpha-toxin

Human skin keratinocytes HaCat attacked by Staphylococcus aureus α-toxin showed a transient drop of cellular ATP levels whereas in toxin-perforated bovine mammary epithelial cells (BMEC), the ATP levels dropped more slowly. Morphologically, during the ATP level depletion, HaCat cell developed a spacious intracellular vacuole together with the transient influx of trypan blue. WST-1 signal, which tested the function of mitochondrial enzyme in viable cells, also decreased concomitantly. On the other hand, BMEC excluded trypan blue and vacuolation was not observed throughout the experiment. We conclude that mammary epithelial cells resist the toxin better than keratinocytes. This is the first report showing that α-toxin enhances transient membrane permeability to large molecules, temporary vacuole formation and the transient defect of mitochondrial enzyme in viable cells without cell lysis.     

Detection of Staphylococcus aureus in bulk tank milk using modified Baird-Parker culture media.

The purpose of this project was to evaluate the use of 2 selective/differential culture media for detecting Staphylococcus aureus in bulk tank milk. One medium was Baird-Parker agar base supplemented with egg york tellurite emulsion and acriflavine. The other medium was Baird-Parker agar base supplemented with rabbit plasma/bovine fibrinogen and acriflavine. An increased inoculum of bulk tank milk (0.3 mL) was used to enhance the detection of S. aureus in samples containing low numbers of organisms. The sensitivity and specificity for detecting S. aureus in bulk tank milk were 94.8% and 100%, respectively, using Baird-Parker agar base supplemented with egg yolk tellurite emulsion and acriflavine, and 89.7% and 100%, respectively, using Baird-Parker agar base supplemented with rabbit plasma/bovine fibrinogen and acriflavine. Both media are practical for detecting S. aureus in bulk tank milk and monitoring its spread in lactating dairy herds in Alberta.     

An outbreak of methicillin-resistant Staphylococcus aureus skin infections resulting from horse to human transmission in a veterinary hospital

There are increasing reports of methicillin-resistant Staphylococcus aureus (MRSA) infection and colonization in horses and evidence that MRSA can be transmitted between horses and humans. The objective of this study was to investigate reports of skin infection in personnel working with a foal with community-associated MRSA colonization and subsequent infection. Clinical diagnostic specimens were collected from individuals reporting skin lesions following contact with the affected foal. Nasal and groin screening swabs were collected from other veterinary personnel that attended a voluntary screening clinic. MRSA skin infections were identified in three neonatal intensive care unit personnel. Nasal colonization was subsequently identified in 10/103 (9.7%) other veterinary hospital personnel. Isolates were indistinguishable by pulsed field gel electrophoresis, classified as Canadian epidemic MRSA-5, possessed SCCmecIV, were negative for the Panton-Valentine leukocidin and were multidrug resistant. Transmission to veterinary personnel despite short-term contact with standard protective barriers highlights the potential importance of MRSA as an emerging zoonotic pathogen, and indicates that further evaluation of interspecies transmission of MRSA and means to prevent zoonotic infection are required.     

Characteristics of Staphylococcus aureus isolated from lesions of horses.

Seventy-six Staphylococcus aureus strains isolated from various lesions of horses were characterized. All of the 76 strains were identified as biotypes B (38.2%) and C (61.8%). Of 55 strains tested, 42 (76.4%) were differentiated into 7 coagulase types. Coagulase types V and VII were predominant in the metritis strains. Coagulase type II was found most frequently in the strains from phlegmon, dermatitis, sinusitis, empyema sinus, and nasal catarrh. Forty-two (55.3%) of the 76 strains were differentiated into 24 phage patterns. Twenty (58.8%) of 34 typable strains from metritis were lysed by the human group I phage 52, and group II phages 3A, 3C, 55 and 71. Forty-five (59.2%) of the 76 strains were resistant to 1 or more of 6 antibiotics. Strains resistant to penicillin G, irrespective of source, were most frequent (95.6%). Forty (93.0%) of 43 strains resistant to penicillin G alone or in combination with other antibiotics produced beta-lactamase. Only 8 (10.5%) of the 76 strains produced enterotoxins A (n = 2), B (n = 1) or C (n = 5), and they all were isolated from metritis. Only 1 strain isolated from phlegmon and 2 from metritis produced exfoliative toxin (ET) and toxic shock syndrome toxin-1 (TSST-1), respectively. The latter 2 strains also produced enterotoxin C. The results of the present study showed the first evidence of the presence of both ET- and TSST-1-producing S. aureus isolated from horses.     

Lack of growth of heat-shocked Staphylococcus aureus surviving cells in common media and enterotoxin release.

The ratio of the different types of S. aureus surviving cells immediately after sub-lethal heat treatment (52 degrees C for 15 min) was studied. A high ratio of cells which are unable to form colonies even in a common medium such as TSA was observed. This fact has not been reported previously for common media through recovery after heat shock has been widely studied. After the treatment, staphylococcal protein A, but no enterotoxin, was seen to be released into the extracellular environment.     

Influence of milking technique and lactation on the bovine teat by means of ultrasonographic examination

The teats of Brown Swiss and Austrian Simmental cows, divided into two groups, one milked by means of an automatic milking system, the other using a conventional milking parlour, were examined monthly by ultrasonography. Aim of the study was to compare the effects of two different milking machines upon the structures of the bovine teat canal and wall by ultrasonography and thereby evaluate ultrasonography as a research tool for visualisation of different influences on the bovine teat. Length and thickness of the teat canal and teat wall thickness were measured and analysed. During lactation, teat canal length and thickness increased in both groups, teat canal length decreased in conventional milked cows at the end of lactation. Shorter and narrower teat canals were observed in automatic milked cows. Differences between the groups in teat canal length and thickness were determined in early lactation. During lactation teat wall thickness showed a slight increase. Automatic milked cows displayed thinner teat walls than cows milked in the milking parlour. Teat morphology was influenced by the number and duration of lactations, milk yield, quarter of the udder and time and date of examination. It was concluded that the effect of the two different milking machines caused significant differences in bovine teat morphology and that ultrasonography proved to be an appropriate method for visualising influences of the milking technique on the bovine teat.     

Antimicrobial susceptibility of Staphylococcus aureus and Staphylococcus pseudintermedius isolated from various animals

This study characterized the antimicrobial susceptibility of 221 Staphylococcus aureus isolated from various species, and 60 canine Staphylococcus pseudintermedius isolated from 1986 through 2000 at the Western College of Veterinary Medicine (WCVM). Resistance of S. aureus was most common to penicillin (31%) and tetracycline (14%); resistance of S. pseudintermedius to penicillin was present in 8% and to tetracycline in 34% of isolates. Resistance to trimethoprim/sulfamethoxazole was only seen among S. pseudintermedius, and there was no resistance to amoxicillin/clavulanate, ampicillin/sulbactam, cephalothin, amikacin, gentamicin, enrofloxacin, chloramphenicol, or rifampin among any isolate. Inducible clindamycin resistance was found in both S. aureus and S. pseudintermedius, highlighting the need for careful interpretation of culture and susceptibility test results. There were significant differences in the minimum inhibitory concentrations of penicillin, ciprofloxacin, enrofloxacin, clindamycin, erythromycin, chloramphenicol, and tetracycline between avian, bovine, equine, and porcine isolates.     

一株鸡源猪葡萄球菌的分离与鉴定

为进一步研究引起蛋鸡产蛋下降的病原菌的致病机理,本研究从患有输卵管炎的蛋鸡输卵管中分离到一株革兰氏阳性球菌,通过培养形态观察和生化试验进行初步鉴定,并采用PCR扩增其16S rRNA基因序列进行测序,与GenBank中登录的已知序列进行比对分析,结果显示该菌与猪葡萄球菌(Staphylococcus hyicus)在进化关系上的距离最近,同源性达到99.5％以上,表明该分离株为S.hyicus.并通过小鼠毒力试验和蛋鸡回归试验对该分离株的致病力研究,表明该菌对小鼠无致病性,但可以引起生殖系统病变,导致产蛋能力下降.