Phenotypic and genotypic properties of Staphylococcus aureus subsp. anaerobius isolated from lymph node abscesses of goats

In the present study 20 staphylococci isolated from lymph node abscesses of 19 goats of two herds in Western Poland could be identified as Staphylococcus aureus subsp. anaerobius. All 20 strains grew under microaerobic conditions, were negative in the catalase test, showed the typical phenotypic properties of 5. aureus and could genotypically be identified by a positive sa442, 235 rDNA, nuc, coa and spa PCR reaction. The variable regions of the coa and spa gene of the 20 strains appeared with uniform amplicon sizes, respectively. All 20 strains were negative for 12 additionally investigated enterotoxin encoding genes, tst and ssl7 and positive for the gene cap8. Identical properties could be observed for S. aureus subsp. anaerobius DSM 20714. Amplification and sequencing of kat gene of a single Staphylococcus aureus subsp. anaerobius strain of the present study and S. aureus subsp. anaerobius DSM 20714 revealed a complete identity of the kat sequences of both strains and a katB sequence obtained from GenBank (AJ000471). The bacteria were additionally investigated for relatedness by macrorestriction analysis of chromosomal DNA with subsequent pulsed-field gel electrophoresis (PFGE), yielding, corresponding to the above mentioned PCR results, identical PFGE patterns for all 20 Staphylococcus aureus subsp. anaerobius strains isolated in Western Poland and the S. aureus subsp. anaerobius reference strain DSM 20714.This indicates the clonal identity of the strains isolated in Western Poland and the S. aureus subsp. anaerobius reference strain. The route of infection of the two herds in Western Poland with a bacterial clone originally isolated in Spain remains unclear.     

Staphylococcus aureus isolated from poultry in Australia. I. Phage typing and cultural characteristics

The phage typing and cultural characteristics of 574 strains of S. aureus of poultry origin in Australia were examined. With the avian phage set of Shimizu (1979) it was possible to type 74.2% of strains. A number of significant variations in the phage typing patterns of Australian strains compared to those reported from Japan and Europe were observed. A lower proportion of Australian strains were of avian phage group I and a higher proportion of group III. A high proportion of strains were of mixed lytic groups. No locally isolated phages were able to increase significantly the percentage of typeable strains, although four local phages appeared to be of greater value for phage typing poultry strains of S. aureus than some other phages of the avian phage set. The international (human) phage set was of limited value in typing Australian strains of poultry origin although four strains were identified which were indistinguishable from strains of human origin. Using cultural characteristics of the strains in conjunction with phage typing, the Australian strains of S. aureus were assigned to one of three major groups and nine subgroups. A list of typing phages considered to be valuable for use on Australian poultry strains of S. aureus is given.     

Interaction of sub-epithelial connective tissue components with Staphylococcus aureus and coagulase-negative staphylococci from bovine mastitis

Staphylococcus aureus and coagulase-negative staphylococci (CNS) isolated from bovine mastitis were examined for their ability to interact with 125I-labelled fibronectin, fibrinogen and type II collagen. Their relative surface hydrophobicity and production of extracellular capsule were also investigated. Almost all S. aureus strains bound fibronectin (mean value 23%), fibrinogen (mean value 12%) and type II collagen (mean value 16%). CNS bound fibronectin (mean value 6%) and type II collagen (mean value 7%), but not fibrinogen (mean value 2%). The specificity of binding of these proteins to S. aureus strain F1440 and to coagulase-negative Staphylococcus chromogenes strain BO52 was studied by adding an excess of unlabelled proteins. Fibronectin and collagen binding were observed to be specific, varying between 50 and 75%, whereas the specificity of fibrinogen binding to S. aureus strain F1440 was lower (26%). Most of the S. aureus strains (63%) showed very high surface hydrophobicity (autoaggregation) or lower hydrophobicity (29% of the strains) and the rest were hydrophilic. None of the CNS strains autoaggregated, 44% were classified as hydrophilic strains. Hydrophilic strains (except the reference strains) did not show extracellular capsule production. However, the encapsulated (reference) strains showed low binding to these proteins as compared to their unencapsulated variants. Pre-treatment of S. aureus strain F1440 and S. chromogenes strain BO52 with trypsin decreased their fibronectin binding capacity and surface hydrophobicity, whereas pre-treatment with bovine milk (except on collagen binding to strain F1440) did not significantly affect binding to these proteins. These data indicate that S. aureus and CNS isolated from bovine udder infection have the ability to bind to tissue matrix and plasma proteins which may be exposed in the traumatized or toxin-damaged udder epithelial lesions.     

湖南猪源粪肠球菌的分离鉴定及16S rDNA系统进化分析

从湖南各地送检至实验室的临床样本中分离到42株革兰氏染色阳性及过氧化氢酶阴性的球菌,其中78.6%来自于肺脏,菌落形态及染色镜检均与粪肠球菌参考株ATCC 29212相似,兰氏分群鉴定97.6%（41/42）为D群,生化特性符合粪肠球菌特征。16SrDNA测序鉴定显示：42个分离株与同步测序的ATCC 29212同源性在99.2%到100%之间,与所选择粪肠球菌参考序列的同源性在98.7%到100%之间,NCBI在线BLAST分析发现42株均与GeneBank收录的粪肠球菌序列同源性最高。湖南分离株16SrDNA序列与E.faecalis参考序列进化关系与地域分布无关,来源于不同宿主的菌株的16SrDNA变异并不大,而与E.faecium、E.canis、E.avium、E.hirae等4种肠球菌则有多处变异,这些变异区域在这4种肠球菌中却是保守的。     

犬冠状病毒JS1706和JS1712毒株基因组3'端分子特性分析

为了全面了解犬冠状病毒（CCoV）分离毒株JS1706和JS1712基因组3'端主要结构蛋白基因和非结构蛋白基因的分子特征,本研究设计了8组引物进行RT-PCR扩增,产物经测序和拼接后,获得了约8.7 kb基因组片段,该基因组结构及其编码蛋白顺序为5'-S-3abc-E-M-N-7ab-3'。对CCoV JS1706、JS1712株8.7 kb基因组核苷酸序列与α冠状病毒属参考毒株的相同区域核苷酸序列进行比对,结果表明,2个分离株与CCoV Ⅱ型参考毒株相似性最高（83.4%~93.1%）,其次为FCoV Ⅱ型参考毒株（87.1%~87.9%）、TGEV参考毒株（86.1%~86.8%）、CCoV Ⅰ型参考毒株（72.0%~72.1%）和FCoV Ⅰ型参考毒株（67.5%~69.9%）。JS1706、JS1712毒株与同属冠状病毒参考株的结构蛋白S、E、M和N蛋白氨基酸相似性分别为46.4%~95.2%、75.6%~100%、82.8%~99.2%和78.5%~99.7%。说明同属内冠状病毒的S基因变异度大,E、M、N基因相对保守。根据基因组3'端8.7 kb核苷酸序列和S蛋白氨基酸序列相似性比对结果,JS1706和JS1712毒株均与泛嗜型原型株CB/05相似性最高,分别为93.0%~93.1%、94.8%~95.2%,其他结构蛋白包括E、M和N氨基酸序列比对也发现与CB/05株的相似性较高,分别为97.6%~100%、92.4%~93.1%和97.9%。S蛋白氨基酸序列的进一步分析表明,JS1706和JS1712毒株的S蛋白N端有一些特有氨基酸,S蛋白氨基酸序列中没有明显的S1/S2蛋白酶切位点（RRARR）,但在958—963位氨基酸有S2'裂解位点特征基序（KRKYRS）。基于S蛋白氨基酸序列构建的系统发育进化树分析显示,CCoV JS1706和JS1712株与CCoV Ⅱa亚型参考毒株和FCoV Ⅱ型参考毒株聚集形成一个分枝。CCoV JS1706和JS1712株非结构蛋白的编码基因ORF3abc和ORF7,其结构、大小与经典疫苗株INSAVC-1相似,无明显插入、缺失和移码突变。本研究有助于深入了解国内CCoV流行毒株的分子特性,为后续分子流行病学调查、诊断试剂和疫苗研发奠定了基础。     

Serologic cross-reactivity of Australian Moraxella bovis to vaccinal bacterin strains as determined by competitive ELISA

OBJECTIVE: To conduct a serologic survey and define pili antigenic variability via the serologic cross-reactivity of Moraxella bovis isolates from naturally occurring infectious bovine keratoconjunctivitis (IBK) outbreaks in Australia. This project applies to the development of an M bovis pili-based vaccine targeting Australian strains originating from intensive cattle producing regions. PROCEDURE: Ocular swabs were collected from cattle affected with clinical signs of IBK from 25 veterinary practices. Standard criteria were used to identify 70 M bovis. Pure, piliated isolates were evaluated with a modified competitive enzyme-linked immunosorbent assay (ELISA) for cell-bound M bovis pili to determine their serologic cross-reactivity with pili of vaccinal bacterin strains EPP63, FLA64, and SAH 38. RESULTS: Sixty-four percent (45/70) of M bovis isolates demonstrated homologous pili antigens to a vaccinal strain. M bovis isolates homologous to one of the three vaccinal strains were obtained in 77% (34/44) of IBK outbreaks sampled. No IBK outbreak had isolates homologous to more than one vaccinal strain; however, 29% (10/34) of outbreaks with a cross-reacting strain had non-cross-reacting strains as well. CONCLUSION: The similar prevalence of pilus antigen homology to strain FLA64 was observed with isolates derived from NSW, Tasmania, and Victoria, compared with results of prior smaller serologic studies, suggests that the common pilus antigens in M bovis within Australia have been relatively stable over the last 20 years. The prevalence of a limited number of pilus antigens in M bovis suggest that the application of a vaccine containing the bacterial strains EPP63, FLA64, and SAH38 may provide a useful management tool for reducing production losses associated with IBK in Australia.     

In vitro antimicrobial activity of orbifloxacin against Staphylococcus intermedius isolates from canine skin and ear infections

The objective of the study was to evaluate the in vitro activity of orbifloxacin against Staphylococcus intermedius strains isolated in France from canine skin and ear infections. The minimum inhibitory concentrations (MICs) of orbifloxacin against 240 field S. intermedius isolates (69 skin and 171 ear isolates) ranged from 0.016 to 8 mg l(-1), with MIC50 and MIC90 equal to 0.5 and 1 mg l(-1), respectively. Only one strain, a pyoderma isolate was resistant (MIC=8 mg l(-1)). Orbifloxacin was tested at different concentrations for killing rate against five isolates obtained from pyoderma cases and against a reference strain (Staphylococcus aureus ATCC 29213). Orbifloxacin expressed a concentration-dependent bactericidal activity against the S. aureus reference strain, but a time-dependent bactericidal activity against S. intermedius. Orbifloxacin induced bactericidal effect against the S. intermedius strains tested with concentrations equal to or two times MIC.     

Isolation of Mycoplasma capricolum-like strains from chickens

Members of the genus Mycoplasma infect a wide range of hosts, but individual Mycoplasma species tend to exhibit a considerable degree of host specificity. We characterized Mycoplasma strain 700, isolated from a kidney of a layer hen in Spain and Mycoplasma strains ULB-A and ULB-B, isolated from the air sac and from the bile of stunted broiler chickens in Slovenia. The serologic examination showed that these three strains are antigenically unrelated to all of the recognized Mycoplasma species of avian origin, but closely related to the ruminant mycoplasma Mycoplasma capricolum subspecies capricolum (M. capricolum). The comparison of their 16S rRNA gene sequences with the sequence of M. capricolum (California kid) revealed 99.66% sequence identity for the strain 700 and 99.59% identity for strains ULB-A and ULB-B. Moreover, the predicted DnaK sequences of the M. capricolum-like strains, isolated from chickens, were identical to DnaK sequences of M. capricolum. Comparison of their dnaK gene sequences with M. capricolum showed 99.64% sequence identity for strain 700 and 99.27% identity for strains ULB-A and ULB-B. In the flock from which M. capricolum-like strains ULB-A and ULB-B were isolated, the majority of chickens (83% of the chickens examined) raised antibodies reacting with M. capricolum antigens. Notably, the infection of chickens with M. capricolum-like strains represents an unusual exception to the range of Mycoplasma species host specificity.     

陕西省部分地区猪流行性腹泻病毒S、M和N基因的遗传变异分析

为了解陕西省部分地区猪流行性腹泻病毒(PEDV)的遗传和变异情况,采集陕西省部分地区规模化猪场的5份疑似PEDV感染的猪小肠内容物,进行PEDV S、M和N基因的RT-PCR扩增,并对扩增产物进行序列测定和遗传变异分析。结果表明,5份病料均能扩增出PEDV S、M和N基因,5株病毒分别命名为SXSL、SX-BJ、SX-YL、SX-WN和SX-HZ株。序列分析表明,5株毒株之间的S、M和N基因核苷酸序列的同源性分别为96.7%~99.8%、98.4%~100%和97.2%~99.9%;氨基酸序列的同源性分别为97.4%~99.9%、98.2%~100%和98.2%~100%。该5株病毒与中国疫苗株CV777的S、M和N基因核苷酸序列的同源性分别为93.9%~99.8%、98.1%~100%和95.3%~99.9%,氨基酸序列的同源性为93.6%~99.9%、96.2%~100%和98.2%~100%。遗传进化分析结果显示,5个陕西分离株的S基因与中国疫苗株CV777亲缘关系较远,与近年来中国株、日本株以及韩国株亲缘关系较近。SX-SL株、SX-BJ株和SX-YL株的M和N基因与中国疫苗株CV777亲缘关系较近,且与中国株CHGD-01亲缘关系密切。SX-WN株和SX-HZ株的M和N基因与中国疫苗株CV777亲缘关系较远。该5株病毒的S基因以及SX-WN株和SX-HZ株的M基因和N基因变异程度较大,而SX-SL株、SX-BJ株和SX-YL株三个流行株均与中国株CHGD-01亲缘关系密切,并且与近年在陕西省流行的PEDV也不完全相同。     

华东地区部分猪源大肠杆菌K88黏附素的生物学特性

近年来，从华东地区患腹泻仔猪中分离到一些表达K88菌毛的大肠杆菌，这些菌株只与K88a因子单抗反应，而不与b、c、d因子单抗反应。通过K88常规血清交叉吸收试验、SDS-PAGE、Western印迹，表明这些菌株不仅与K88ac参考菌株C83907制备的c因子血清反应，而且与以分离株SEC586制备且经K88ab、K88ac、K88ad参考菌株吸收后的血清也反应。对分离株SEC586、SEC464的K88主要亚单位结构基因faeG的克隆、测序，发现该基因由846对核苷酸组成，编码菌毛主要亚单位的262个氨基酸及21个氨基酸的信号肽，比国外报道的K88ac FaeG亚单位（263个氨基酸）少了1个氨基酸，比K88ab、K88ad（265个氨基酸）少了3个氨基酸。SEC586、SEC464菌株的FaeG亚单位氨基酸序列的同源性为97．7％，它们与K88ac的同源性为94．7％和96．2％；与K88ab的同源性为90．1％和91．2％；与K88ad的同源性为87．0％和88，6％。结果表明，新分离的K88ac大肠杆菌黏附素主要亚单位已发生了部分变异。     

Comparison of three commercial rapid agglutination test kits for identification of coagulase positive staphylococci from foods and animals.

Three rapid agglutination assays for the identification of Staphylococcus aureus Monostaph (Bionor A/S, Skien, Norway), Staphyslide-Test (BioMerieux, Lyon, France) and Staph-Rapid-Test (Roche, Basel, Switzerland), were compared. A total of 104 Gram-positive, catalase positive cocci were tested: Nineteen Staphylococcus reference strains comprising 15 spp. (4 strains were coagulase positive), and 7 Micrococcus reference strains comprising 4 spp.; 22 food isolates comprising 13 S. aureus, 8 coagulase positive Staphylococcus spp., and 1 Micrococcus sp.; 56 animal isolates comprising 11 S. aureus, 9 S. hyicus subsp. hyicus, 2 S. intermedius, 15 coagulase positive and 19 coagulase negative Staphylococcus spp. Totally 54 strains were coagulase positive. Considering agglutination of a coagulase positive strain as a correct identification, Monostaph, Staph-Rapid-Test, and Staphyslide-Test correctly identified 52 (96.3%), 47 (87.0%) and 48 (89.0%) of the coagulase positive staphylococci, respectively. Monostaph, Staph-Rapid-Test and Staphyslide-Test showed 1 (2.0%), 4 (8.0%) and 4 (8.0%) false positive reactions respectively. Monostaph, Staph-Rapid-Test and Staphyslide-Test gave 0 (0.0%), 6 (5.8%) and 7 (6.7%) non-interpretable reactions, respectively. Monostaph may be a good alternative to the tube-coagulase test for rapid and reliable identification of coagulase positive staphylococci from both food and veterinary sources. However, false negative reactions may occur with coagulase positive strains of S. hyicus subsp. hyicus and S. intermedius.     

Evaluation of three slide agglutination tests for rapid identification of Staphylococcus aureus.

Three slide agglutination tests for identification of Staphylococcus aureus were compared. The agglutination tests used for evaluation were Staphaurex (Wellcome Diagnostics), Staphyslide-Test (BioMerieux), and ANI S. aureus TEST (Ani Biotech Oy). A total of 347 isolates were analyzed, including 288 strains of S. aureus, 49 of S. epidermis, 11 of S. intermedius, 12 strains of other staphylococci and 14 non-staphylococcal strains. One hundred of the S. aureus strains were isolates from cases of food poisoning, 129 from mastitis and 59 from other clinical cases. The sensitivities of the tests were also compared using diluted suspensions of S. aureus strains and with purified Protein A dilutions. The results showed that the sensitivities of the tests were 98.6%, 97.9% and 99.0% for Staphaurex, Staphyslide-test and ANI S. aureus TEST, respectively. The specificities were 100% for the Staphyslide test and 98.8% for both the ANI S. aureus TEST and the Staphaurex test. The sensitivities measured with diluted S. aureus strain suspensions and Protein A solutions were equal with the Staphaurex and ANI S. aureus TEST. All the agglutination tests studied proved to be practical, easy to use and accurate for the rapid identification of S. aureus strains from culture isolates.     

奶牛乳房炎源金黄色葡萄球菌溶原性噬菌体的诱导及其裂解能力分析

为研究奶牛乳房炎源金黄色葡萄球菌中溶原性噬菌体的存在情况及其裂解能力,对采集的45份临床型乳房炎乳样进行金黄色葡萄球菌分离及PCR鉴定,通过丝裂霉素C对分离出的金黄色葡萄球菌进行诱导溶原性噬菌体,对诱导出的噬菌体进行纯化并测定其裂解能力.结果表明分离到了16株金黄色葡萄球菌,分离率为35.56％(16/45);诱导出了...     

广西部分猪场猪流行性腹泻病毒S基因克隆及序列分析

采集广西25个猪场的病料,对猪流行性腹泻病毒(PEDV)阳性组织样品进行全S基因扩增.将41株全S基因进行序列比对及遗传进化分析,41株PEDV的S基因之间核苷酸同源性为94.8％～100％,与参考毒株核苷酸同源性为89.3％～99.4％.S基因进化树图谱显示,广西当前流行的PEDV可分为2个谱系.Attenuated...     

Secreted antigens as virulence-associated markers in Staphylococcus aureus strains from rabbits

Western blot analysis was performed from the culture supernatant of 59 rabbit Staphylococcus aureus strains, classified as high and low virulence strains according to their epidemiological behaviour in commercial rabbitries, bio-, phage- and RAPD-type. Fourteen extracellular antigen bands (A-N) were recognised using sera of rabbits immunised with washed, viable high virulence S. aureus bacteria. Eleven of these bands were found in high virulence as well as in low virulence strains. The band A, approximately 78 kDa, was not seen in any of the 27 high virulence strains, except for one strain which was also typical in other aspects, was detected in all, but one of the low virulence strains. The M and N bands with molecular masses of approximately 29 and 27 kDa, respectively, were recognised in all high virulence strains except for the atypical strain, but in none of the low virulence strains. This indicates that the latter two antigens may be virulence-associated markers for S. aureus strains from rabbits.     

Characterization of two proteins of Staphylococcus aureus isolated from bovine clinical mastitis with homology to glyceraldehyde-3-phosphate dehydrogenase

Staphylococcus aureus is the most common causative agent of bovine mastitis and vaccines developed to control this disease showed limited protection due in part to the lack of common antigens among the mastitis isolates. We isolated and identified two genes encoding proteins with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity from a S. aureus strain isolated from bovine clinical mastitis. The GapB and GapC proteins share considerable homology to the GapB and GapC products of human strains of S. aureus. These two proteins could be distinguished by their different GAPDH activities and binding to bovine transferrin properties. Both gapB and gapC genes were conserved in 11 strains tested, and the GapC protein was present on the surface of all S. aureus strains.     

Lytic activities, protein profiles and morphologic characteristics of new bacteriophages isolated from canine and human Staphylococcus aureus strains.

The lytic activity, protein profile and morphology of five newly isolated phages from canine Staphylococcus aureus strains and one from a human S. aureus strain were compared with those of selected phages in the international phage sets (IPS). Five canine phages lysed 57 (76.0%) of 75 canine isolates of Staphylococcus aureus from Nigeria at routine test dilution (RTD) while 34 (IPS) phages typed only 31 (41.3%) strains at RTD or/and 100-RTD. The new human phage lysed 11 (14.7%) of 75 strains isolated from human diarrhoea. The new phages were readily propagated, specific in activity and stable during storage at 4 degrees C. Prominent proteins detected by SDS-PAGE indicated similarities between some of the phages but one canine phage was distinctly different, as was its morphology which was an isometric head with a short tail compared to oval heads and long tails which characterized others. IPS phages in the same serologic group had similar protein profiles but no correlation was observed with lytic groups. The use of protein profile and electron micrographs allowed classification of the phages into serogroups. It is concluded that the newly isolated canine phages could be very useful in typing Nigerian canine strains of S. aureus.     

Genetic characterisation of Australian strains of porcine circovirus types 1 and 2

OBJECTIVE: As post-weaning multi-systemic wasting syndrome (PMWS) has not been identified within Australia, to determine if the absence of disease was associated with genetic differences between the strains of porcine circovirus (PCV) present in Australia and those from countries in association with PMWS. DESIGN: Pig tissues were obtained from weaned pigs found dead or presenting with clinical signs of illthrift and also from neonatal pigs with congenital tremors and used as a source of virus DNA for sequence analysis. PROCEDURE: DNA was extracted from the tissues and PCV detected by polymerase chain reaction (PCR). PCR with PCV type-specific primers was used to amplify the entire genome from selected tissues. The genomes of three strains of PCV1 and seven strains of PCV2 from three Australian states were sequenced and subjected to phylogenetic analysis using standard procedures. RESULTS: The three Australian PCV1 strains had 98 to 99% nucleotide identity to strains in other countries and the seven Australian PCV2 strains had 94 to 99% identity to PCV2 strains in other countries where PMWS has occurred. Six of the seven Australian PCV2 strains were genetically similar to each other, while the seventh was more distantly related. There were no consistent differences in the predicted amino acid sequence of the Australian strains of PCV2 and strains associated with PMWS in other countries. CONCLUSION: There were no consistent differences between Australian strains of PCV and those that have been associated with PMWS in other countries and it appears likely that other factors are responsible for the absence of PMWS in Australia.     

Isolation of a monoclonal antibody with specificity for commonly employed vaccine strains of Newcastle disease virus

A monoclonal antibody, AVS-I, was produced from a hybridization of murine myeloma cells and splenocytes from mice immunized with the La Sota strain of Newcastle disease virus (NDV). The hybridoma producing AVS-I, selected from 184 NDV-positive supernatants, is one of two supernatants that reacted exclusively with lentogenic strains in an indirect enzyme-linked immunosorbent assay. AVS-I can also be assayed by hemagglutination-inhibition (HI), which was used to test selected reference avian paramyxovirus (PMV) strains of types 1 to 3. NDV vaccines La Sota and B1 and field isolates from chickens, turkeys, pigeons, and cockatoos were also used as antigens. AVS-I had a high binding affinity for all La Sota and B1 strains, including vaccines. The antibody bound with a lower titer to the Australian Queensland V4 and Ulster strains, but it did not bind to the F strain, a lentogenic strain from England. AVS-I was HI-negative against the other PMV reference strains. AVS-I may be valuable for identifying field isolates antigenically similar to La Sota and B1 and rapidly differentiate those vaccine strains from more virulent viruses.     

Induced staphylococcal infections in the bovine mammary gland

In a study to develop and define a practical model of bovine mastitis caused by Staphylococcus aureus, induced infections were attempted in 203 bovine mammary glands of 41 cows, using 12 strains of S aureus. Approximately 100 colony-forming units of S aureus in saline solution were injected after milking, and milk samples were collected daily from test glands for 14 days to monitor the progress of infections and inflammatory responses. Relationships were examined for cow-related factors and for various characteristics of the strains of S aureus used to the development of a persistent intramammary infection. A dairy cow that was useful in this model was defined as follows: (1) the 2nd to 7th month in the 1st to 5th lactation; (2) producing milk from all mammary glands that contained less than 6 x 10(5) somatic cells/ml; and (3) having mammary glands that were free of any primary mastitis pathogen, as well as micrococci and Corynebacterium bovis. From the present study, it was not possible to define clearly a strain of S aureus which would be useful in the model, but 5 strains of S aureus were identified as being capable of producing persistent subacute infections with a high degree of repeatability.