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Transmissibility of Mycoplasma mycoides subsp. mycoides infection from experimentally inoculated goats to other goats and calves was studied. Eight goats and six calves were housed in an 18 m2 room. Six of the goats were inoculated endobronchially with strain D44 isolated from a natural case of polyarthritis in Ontario. These six goats died within a week of Mycoplasma septicemia. The two contact goats or the six calves never showed signs of disease and M. mycoides subsp. mycoides was not recovered from these animals. The contact goats and four calves were killed 25 days after exposure. They were all seronegative, M. mycoides subsp. mycoides was not recovered at necropsy and none had pathomorphological changes attributable to this Mycoplasma. The two remaining calves were inoculated endobronchially with 10(9) CFU of strain D44 and observed for 20 days. They never showed signs of disease and did not have significant lesions at necropsy. Both developed a significant serological response to M. mycoides subsp. mycoides, although this organism was not recovered during the experimental period or at necropsy. This study did not provide evidence for transmission of M. mycoides subsp. mycoides from endobronchially inoculated goats to contact goats or calves and endobronchially inoculated calves did not develop pneumonia. This would suggest that the infection of the goat population in Canada with this pathogen would not be a significant threat to the cattle population.

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Mycoplasma ovipneumoniae is a primary pathogen.

Epizootic pneumonia of bighorn sheep is a devastating disease of uncertain etiology. To help clarify the etiology, we used culture and culture-independent methods to compare the prevalence of the bacterial respiratory pathogens Mannheimia haemolytica, Bibersteinia trehalosi, Pasteurella multocida, and Mycoplasma ovipneumoniae in lung tissue from 44 bighorn sheep from herds affected by 8 outbreaks in the western United States. M. ovipneumoniae, the only agent detected at significantly higher prevalence in animals from outbreaks (95%) than in animals from unaffected healthy populations (0%), was the most consistently detected agent and the only agent that exhibited single strain types within each outbreak. The other respiratory pathogens were frequently but inconsistently detected, as were several obligate anaerobic bacterial species, all of which might represent secondary or opportunistic infections that could contribute to disease severity. These data provide evidence that M. ovipneumoniae plays a primary role in the etiology of epizootic pneumonia of bighorn sheep.

Contagious caprine pleuropneumonia (CCPP) is a major threat to goat farming in parts of Africa and Asia. It classically causes acute high morbidity and mortality early in infection, but little is known of its long term epizootiology and course. In this study, 10 goats were inoculated with Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae) and then mixed with 15 goats for contact transmission. The disease course was monitored in each goat for 56–105 days, whereafter the goats were killed and necropsied. Varying features signifying infection occurred in altogether 17 goats (7 inoculated, 10 in-contact). Clinical signs were severe in 8 goats but no fatalities occurred. Only 6 goats had serum antibody titres against M. capripneumoniae in ELISA. Fourteen goats (5 inoculated, 9 in-contact) had chronic pleuropulmonary lesions compatible with CCPP at necropsy and 7 of those showed M. capripneumoniae antigen in the lung by immunohistochemistry. Neither cultivation nor PCR tests were positive for the agent in any goat. The results indicate that the clinical course of CCPP in a flock may be comparatively mild, M. capripneumoniae-associated lung lesions may be present at a late stage of infection, and chronic infection may occur without a significant serological response.

Ten mycoplasmas were isolated from 130 nasopharyngeal swabs from thoroughbred horses with acute respiratory disease and three from 198 apparently normal horses. Two mycoplasmas were isolated from 21 tracheal swabs taken at necropsy. These mycoplasmas, together with six isolated from the equine respiratory tract by other workers, were subjected to biochemical and serological tests. Other properties examined in certain representative strains were appearance under the electron microscope, ability to adsorb or agglutinate the erythrocytes of various animal species and the electrophoretic pattern of the cell proteins. On the basis of these test, mycoplasmas from the equine respiratory tract were divided into seven species. Three species belonged to the genus Acholeplasma, members of which do not require sterol for growth, and were identified as A. laidlawii, A. oculi (formerly A. oculusi) originally isolated from the eyes of goats, and a recently named species A. equifoetale, previously isolated from aborted equine fetuses. Of the four sterol-dependent Mycoplasma species, one was indentified as M. pulmonis, a common rodent pathogen. Another cross-reacted serologically with M. felis and should probably be classified as that species. The other two species probably represent new species peculiar to the horse. One of these, represented by the strains N3 and N11, ferments glucose and is serologically distinct from 19 recognized species of glucose-utilizing mycoplasmas and from two species which do not metabolize either glucose or arginine. The other species, represented by four strains, hydrolyses arginine and, because it is serologically distinct from all the named arginine-hydrolysing Mycoplasma species, the name M. equirhinis sp.nov. is proposed for it. Of the seven species, only M. pulmonis and the glucose-utilizing species represented by N3 and N11 were found exclusively in horses with acute respiratory disease. A. oculi was isolated from an apparently normal horse. The other four species were found in normal horses as well as those with respiratory disease, although three out of the four strains of M. equirhinis were from sick horses.

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Immunoblotted protein samples from several strains of Mycoplasma hominis and from one strain of Mycoplasma arginini each contain a polypeptide of a molecular mass of 95,000 to 105,000 Da which binds immunoglobulin nonimmunologically. Immunoblots from these organisms were probed with alkaline phosphatase-conjugated goat anti-rabbit immunoglobulin, conjugated goat immunoglobulin G (IgG) Fab fragments, and conjugated goat IgG Fc fragments. The polypeptide bound the goat anti-rabbit molecules and the Fab fragments but not the Fc fragments. These reactions could be blocked with nonimmune unconjugated goat IgG and unconjugated human IgM. Controls probed with alkaline phosphatase alone did not stain. Binding of the conjugated preparations to whole mycoplasmal cells was dependent on concentrations of both conjugate and cells for the goat anti-rabbit preparation and for Fab. The mycoplasmal polypeptide may be a light-chain-specific reactant.

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Samples of cervico-vaginal mucus from 633 animals from 110 herds were cultured and yielded the following mycoplasmas: T-strain--88: Mycoplasma bovigenitalium--79, Mycoplasma spp. (Leach Group 7)--7, Acholeplasma laidlawii--4, Mycoplasma bovirhinis--2 and one not typable. Uterine exudates and endometrial scrapings from 80 infertile cows in two herds were examined. Four animals were positive, M. bovigenitalium was isolated three times, A. laidlawii and Mycoplasma arginini once each. Sixty-five normal uterine contents from pregnant cows were examined, one yielded M. bovigenigalium and the same organism was recovered from the fetal kidney. T-strain mycoplasma, M. bovigenitalium and other Mycoplasma spp. appear to be a part of the normal flora of the cervico-vaginal region of clinically normal one and two year old bred heifers in Alberta and Saskatchewan. Although M. arginini was not recovered from the cervico-vaginal region, a single recovery was made from the uterus of an infertile cow.

In order to study the pathomorphology and immunohistochemistry of peste des petits ruminants, four goats and two sheep were inoculated intranasally with the Malig-Yemen strain of peste des petits ruminants virus. The animals developed fever, nasal discharge, oral erosions, cough and diarrhea. One goat and one sheep died and one moribund goat was killed. Three animals survived the infection. At necropsy, erosive stomatitis, pneumonia and gastroenteritis were found. Histopathologically the pneumonocytes and epithelial cells of the ileum had eosinophilic cytoplasmic and nuclear inclusions. By an indirect immunoperoxidase method, the nuclei and cytoplasm of the ileal epithelial cells of one goat contained positively (brown) stained antigen, which corresponded to viral nucleocapsids by electron microscopy. Virus appeared to be released through the microvilli of the epithelial cells. We also confirmed the formation of giant cells due to peste des petits ruminants virus.

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Bronchopneumonia is a population-limiting disease in bighorn sheep in much of western North America. Previous investigators have isolated diverse bacteria from the lungs of affected sheep, but no single bacterial species is consistently present, even within single epizootics. We obtained high-quality diagnostic specimens from nine pneumonic bighorn sheep in three populations and analyzed the bacterial populations present in bronchoalveolar lavage specimens of seven by using a culture-independent method (16S rRNA gene amplification and clone library analyses). Mycoplasma ovipneumoniae was detected as a predominant member of the pneumonic lung flora in lambs with early lesions of bronchopneumonia. Specific PCR tests then revealed the consistent presence of M. ovipneumoniae in the lungs of pneumonic bighorn sheep in this study, and M. ovipneumoniae was isolated from lung specimens of five of the animals. Retrospective application of M. ovipneumoniae PCR to DNA extracted from archived formalin-fixed, paraffin-embedded lung tissues of historical adult bighorn sheep necropsy specimens supported the association of this agent with bronchopneumonia (16/34 pneumonic versus 0/17 nonpneumonic sheep were PCR positive [P < 0.001]). Similarly, a very strong association was observed between the presence of one or more M. ovipneumoniae antibody-positive animals and the occurrence of current or recent historical bronchopneumonia problems (seropositive animals detected in 9/9 versus 0/9 pneumonic and nonpneumonic populations, respectively [P < 0.001]). M. ovipneumoniae is strongly associated with bronchopneumonia in free-ranging bighorn sheep and is a candidate primary etiologic agent for this disease.

Five arginine-utilizing, nonglycolytic Mycoplasma species were compared serologically by two dimensional immunoelectrophoresis. The survey included: Mycoplasma hominis, Mycoplasma arthritidis, Mycoplasma gateae, Mycoplasma gallinarum, and three strains of Mycoplasma arginini. Although the M. arginini strains showed strong cross-reactions, each strain produced a different antigenic profile which was distinct and which indicated significant antigenic differences between strains. The M. arginini strains showed strong relationships with M. gateae; lesser cross-reactions were observed with other strains. Common antigens were demonstrable in the entire group. When all these species were compared with a glycolytic species, Mycoplasma gallisepticum, no cross-reactions were observed. Two-dimensional immunoelectrophoresis was a far more sensitive technique than double-immunodiffusion since as many as 20 components could be resolved.

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A combination of quantitative immunoelectrophoresis and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis was used to determine location and molecular weights of surface membrane antigens of four strains of Mycoplasma arginini. Two major surface antigens were identified for M. arginini by absorption of antiserum with whole cells: one surface antigen was strain specific, electrophoretically fast, and prominently located on the surface, whereas the other surface antigen was common to the four strains and of intermediate electrophoretic mobility. Three of the four strains of M. arginini (G-230, 23243, and 27389) possessed immunologically strain-specific antigens which did not cross-react, whereas the leonis strain lacked an immunologically detectable unique surface antigen. A monospecific antiserum prepared against immune precipitates of the strain-specific antigen of strain G-230 detected three polypeptides of 74,000, 44,000, and 17,000 daltons in SDS-polyacrylamide gels of membrane preparations. All four strains shared the common surface antigen which appeared considerably more hydrophobic than the strain-specific surface antigen because it could only be demonstrated by charge-shift immunoelectrophoretic conditions (addition of deoxycholate to the nonionic detergent). Monospecific antiserum to the common antigen of strain G-230 reacted with all four M. arginini strains, but did not react with two other arginine-utilizing species, and recognized three polypeptides of 40,000, 29,000, and 20,000 daltons in membranes of strain G-230. Whereas the common surface antigen is a likely target for conventional serological reactions used for identification of the species M. arginini, strain-specific antigen cannot fulfill this role but must participate in other surface reactions.

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Two unidentified mycoplasmas, N3 and N11, isolated from the respiratory tract of horses, were found to cross-react with strains of M. mycoides subsp. mycoides in indirect immunofluorescence tests, growth-inhibition tests carried out by the running drop/agar-well method, and in complement-fixation and double immunodiffusion tests. Serologically, the equine mycoplasmas were not completely identical with any of the reference strains of M. mycoides with which they were compared. Their cultural characteristics, ability to digest coagulated serum and casein, and survival at 45 degrees C, however, suggested that they were more closely related to strains of M. mycoides subsp. mycoides, such as Y-goat, which are found in goats, than to strains of that subspecies which are pathogenic for cattle.

We have developed a reverse line blot (RLB) hybridization assay to detect and identify the commonest mollicutes causing cell line contamination (Mycoplasma arginini, Mycoplasma fermentans, Mycoplasma hyorhinis, Mycoplasma orale, and Acholeplasma laidlawii) and human infection (Mycoplasma pneumoniae, Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma parvum, and Ureaplasma urealyticum). We developed a nested PCR assay with “universal” primers targeting the mollicute 16S-23S rRNA intergenic spacer region. Amplified biotin-labeled PCR products were hybridized to membrane-bound species-specific oligonucleotide probes. The assay correctly identified reference strains of 10 mollicute species. Cell cultures submitted for detection of mollicute contamination, clinical specimens, and clinical isolates were initially tested by PCR assay targeting a presumed mollicute-specific sequence of the 16S rRNA gene. Any that were positive were assessed by the RLB assay, with species-specific PCR assay as the reference method. Initially, 100 clinical and 88 of 92 cell culture specimens gave concordant results, including 18 in which two or more mollicute species were detected by both methods. PCR and sequencing of the 16S-23S rRNA intergenic spacer region and subsequent retesting by species-specific PCR assay of the four cell culture specimens for which results were initially discrepant confirmed the original RLB results. Sequencing of amplicons from 12 cell culture specimens that were positive in the 16S rRNA PCR assay but negative by both the RLB and species-specific PCR assays failed to identify any mollicute species. The RLB hybridization assay is sensitive and specific and able to rapidly detect and identify mollicute species from clinical and cell line specimens.

Mycoplasma pulmonis was isolated from the pneumonic lung of a rat. Two groups of mycoplasma-free rats were inoculated, one with a culture of the M. pulmonis strain which had been cloned four times (group A) and the other with a lung homogenate of the rat from which the strain had been isolated (group B). A third group (C) consisted of uninoculated control animals. Each group was kept in strict isolation and allowed to breed so that the progeny was naturally exposed to any pathogens present in the inoculated animals. After different periods of exposure, rats were autopsied, respiratory tracts and inner ears were cultured for mycoplasmas and bacteria, and sera were tested for complement-fixing antibodies to murine mycoplasmas.

In group-A rats, M. pulmonis was consistently isolated from the inner ears or lungs from 50 to 715 days after exposure. Complement-fixing antibody to M. pulmonis was detected 20 days after inoculation, but in the naturally exposed progeny antibody took longer than 50 days to develop. Antibodies to the other known mycoplasmas of murine origin, M. arthritidis and M. neurolyticum, were never found. Purulent otitis interna was consistently found from day 55 onwards, while lung lesions were first observed at 85 days and persisted to 715 days. Pulmonary lesions developed more slowly in inoculated parents than in exposed progeny. Similar results were found in group-B rats, which were examined up to 441 days after inoculation. Uninoculated group-C rats were examined up to 768 days of age, but M. pulmonis was not recovered; of the 54 animals whose serum was tested all were negative to the three species of mycoplasmas, except one which had a titre of 16 with M. pulmonis. Pneumonia, bronchiectasis or lymphoreticular hyperplasia were not seen in any of these control rats. Bacterial respiratory pathogens were not isolated from rats in any of the groups, nor was antibody to Sendai virus detected.

The results suggest that M. pulmonis alone can cause pneumonia and bronchiectasis in rats since mechanical carry-over of another pathogen with the initial cloned inoculum is very unlikely and there was no evidence for the participation of any other rat pathogen. The respiratory disease induced by the cloned culture was comparable with that induced by the lung homogenate, and with the well-known syndrome of chronic respiratory disease and bronchiectasis in the rat.

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Caprine tuberculosis (TB) has increased in recent years, highlighting the need to address the problem the infection poses in goats. Moreover, goats may represent a cheaper alternative for testing of prototype vaccines in large ruminants and humans. With this aim, a Mycobacterium caprae infection model has been developed in goats. Eleven 6-month-old goats were infected by the endobronchial route with 1.5 × 103 CFU, and two other goats were kept as noninfected controls. The animals were monitored for clinical and immunological parameters throughout the experiment. After 14 weeks, the goats were euthanized, and detailed postmortem analysis of lung lesions was performed by multidetector computed tomography (MDCT) and direct observation. The respiratory lymph nodes were also evaluated and cultured for bacteriological analysis. All infected animals were positive in a single intradermal comparative cervical tuberculin (SICCT) test at 12 weeks postinfection (p.i.). Gamma interferon (IFN-γ) antigen-specific responses were detected from 4 weeks p.i. until the end of the experiment. The humoral response to MPB83 was especially strong at 14 weeks p.i. (13 days after SICCT boost). All infected animals presented severe TB lesions in the lungs and associated lymph nodes. M. caprae was recovered from pulmonary lymph nodes in all inoculated goats. MDCT allowed a precise quantitative measure of TB lesions. Lesions in goats induced by M. caprae appeared to be more severe than those induced in cattle by M. bovis over a similar period of time. The present work proposes a reliable new experimental animal model for a better understanding of caprine tuberculosis and future development of vaccine trials in this and other species.

Coxiella burnetii is an intracellular bacterial pathogen that causes Q fever. Infected pregnant goats are a major source of human infection. However, the tissue dissemination and excretion pathway of the pathogen in goats are still poorly understood. To better understand Q fever pathogenesis, we inoculated groups of pregnant goats via the intranasal route with a recent Dutch outbreak C. burnetii isolate. Tissue dissemination and excretion of the pathogen were followed for up to 95 days after parturition. Goats were successfully infected via the intranasal route. PCR and immunohistochemistry showed strong tropism of C. burnetii towards the placenta at two to four weeks after inoculation. Bacterial replication seemed to occur predominantly in the trophoblasts of the placenta and not in other organs of goats and kids. The amount of C. burnetii DNA in the organs of goats and kids increased towards parturition. After parturition it decreased to undetectable levels: after 81 days post-parturition in goats and after 28 days post-parturition in kids. Infected goats gave birth to live or dead kids. High numbers of C. burnetii were excreted during abortion, but also during parturition of liveborn kids. C. burnetii was not detected in faeces or vaginal mucus before parturition. Our results are the first to demonstrate that pregnant goats can be infected via the intranasal route. C. burnetii has a strong tropism for the trophoblasts of the placenta and is not excreted before parturition; pathogen excretion occurs during birth of dead as well as healthy animals. Besides abortions, normal deliveries in C. burnetii-infected goats should be considered as a major zoonotic risk for Q fever in humans.

To evaluate host responses, young goats were inoculated subcutaneously with a genetic deletion mutant (deltapurE201) of Brucella melitensis (n = 6), its virulent parental strain 16M (n = 6), or saline (n = 6). No clinical evidence of brucellosis was seen in any goat. Serum antibody titers peaked at postinoculation day (PID) 14. Bacteria in lymph nodes that drained sites of vaccination reached peak numbers of >10(6) CFU/g in both infected groups at PID 7 and progressively declined to PID 84. At necropsy, bacteria were present in mammary lymph nodes or spleen of 33% of goats given virulent 16M but in none of goats given the purE mutant. Lymphadenitis, most severe in goats given 16M, involved depletion of lymphocytes and germinal centers, proliferation of lymphoblasts, and vasculitis. By PID 28, lymph node architecture was restored; there was marked germinal center formation and medullary plasmacytosis. Brucellar antigens, detected with immunoperoxidase techniques, were prominent in capsular granulomas but not in lymph node cortices. Ultrastructurally, bacteria were found in macrophages (>97%) and small lymphocytes (<3%) but not in large lymphocytes. Bacteria were intact in small lymphocytes but in macrophages were in various stages of degradation. The deltapurE phenotype of deltapurE201 was preserved during infection of goat lymph nodes. Unlike Salmonella spp. purE mutants, strain deltapurE201 may be a candidate for efficacy testing; it produced immune responses, was cleared from visceral tissues, and produced less severe pathologic changes than its wild-type parent.

The virulence of T-mycoplasmas for cattle was tested by examining their ability to produce mastitis in cows. It was found that both virulent and avirulent strains of T-mycoplasmas can be isolated from cattle. All of four strains from pneumonic calf lungs and a strain from a case of bovine kerato-conjunctivitis caused mastitis but only two of four strains isolated from the urogenital tract of cows were virulent. None of the human, simian or canine T-mycoplasmas examined were able to cause mastitis in cattle. However, a bovine strain was found to be capable of causing mastitis in goats. Virulent and avirulent strains from the same and different species contain common antigens detected by the metabolic inhibition test. Pathogenicity could not be shown to be characteristic of any particular serotype. The possibility is raised of some species barrier being responsible for the inability of non-bovine strains to infect cattle.

Clinical, microbiological, serological, histological, and therapeutic aspects of two separate outbreaks of caprine keratoconjunctivitis are described. The disease was characterized by a high rate of contagion, rapid onset, intense lacrimation, conjunctival hyperemia, and corneal opacity with neovascularization. In addition, many of the animals developed respiratory illness during the second epidemic. The only organism consistentlyisolated was Mycoplasma conjunctivae. A total of 23 strains were isolated from 18 inflamed conjunctivae, one normal conjunctiva, and the nasal secretions of four goats with concomitant respiratory illness. The convalescent sera of goats in the first outbreak had neutralizing antibody titers to M. conjunctivae that ranged from 1:32 to 1:256. In the milder second outbreak the antibody titers ranged from 1:4 to 1:32 in animals with only ocular disease and from 1:4 to 1:64 in animals with only respiratory disease. Whereas little change was noted in antibody titers of goats with only localized eye disease, 43% of the goats with respiratory disease showed significant fourfold rises. The histological picture was consistent with acute corneal infection. Animals requiring antibiotic treatment appeared to respond favorably to a combination of oxytetracycline and polymyxin B, but not to penicillin. These findings suggest that M. conjunctivae is one cause of epidemic caprine keratoconjunctivitis.

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Strain Y3343 isolated from a goat with septicemia and polyarthritis was studied. The strain was virulent and induced septicemia, polyarthritis and coagulopathy in two goats. Limulus amebocyte lysate active material was present in plasma, but not in higher titre in inoculated goats. Sonicated mycoplasma material induced a dramatic somatic cell response in the mammary gland of cows and goats and marked clotting of the cows' milk, but it did not clot limulus amebocyte lysate or kill chick embryos. Phenol-water extract clotted limulus amebocyte lysate and induced somatic cell response in cows but not in goats. The phenol-water extract did not kill chick embryos, was not pyrogenic in rabbits or goats, and did not induce generalized Shwartzman reaction or change the leukocyte kinetics in rabbits. It therfore appears that the virulence mechanisms of strain Y3343 can not be explained on the basis of factors with strong endotoxin activity.

Mycoplasma ovipneumoniae is associated with chronic nonprogressive pneumonia in both sheep and goats. Studies concerning its molecular pathogenesis, genetic analysis, and vaccine development have been hindered due to limited genomic information. Here, we announce the first complete genome sequence of this organism.

A serological survey in Mozambique to detect antibodies to Cowdria ruminantium, the etiologic agent of heartwater, revealed a seroprevalence of 8.1% (n = 332) for goats in the northern province of Tete and of 65.6% (n = 326) for goats in the southern provinces. Translocation of 10 serologically negative goats from Tete to farms in the south resulted in two clinical cases of heartwater that were fatal. In addition, four goats seroconverted within the study period of 5 weeks. One goat showed no symptoms. Two goats died of other causes, whereas the remaining goat went missing after 1 week. Experimental needle infections of goats and sheep were conducted to confirm results and to isolate different strains of C. ruminantium. These data indicate that translocation of goats from the north to the south of Mozambique bears a high risk of C. ruminantium infection, which can cause fatal disease.

Small colony (SC) strains of Mycoplasma mycoides subsp. mycoides from contagious bovine pleuropneumonia (CBPP) and from goats were compared with large colony (LC) strains of so-called M. mycoides subsp. mycoides from goats and sheep by means of a cross-protection test in which mice were challenged with M. mycoides subsp. capri. Of 13 LC strains, all gave partial cross-protection, and 11 were shown to be more closely related than four SC strains to subspecies capri. In a further experiment, six SC strains--three from CBPP and three from goats--all gave weak partial cross-protection against subspecies capri.

Background

Widespread in the environment, Staphylococcus spp. infect animals and humans as normal flora or pathogens. By extending our recent report of multi-drug resistant (MDR) S. aureus in dairy goats, this study investigated the staphylococcal infection and characterized the MDR-S. aureus and methicillin-resistant S. aureus (MRSA) isolates collected from goats in 2008 to elucidate the appearance of MRSA in goats and the mastitis associated staphylococcus enterotoxin (SE) types. A total of 555 samples were collected from six goat parts and three environmental sources among four dairy goat farms in southern Taiwan. Coagulase-positive and negative Staphylococcus spp. (CPS and CNS, respectively) were also identified. Furthermore, predominant SE genes of nine enterotoxin genes sea through sej along with antimicrobial resistance and genetic variations were determined.

Results

In total, 137 staphylococcal strains were identified and found predominantly in milk, and in the vagina, anus, and nasal cavity. The most prevalent species was S. lentus, followed by S. aureus, S. epidermidis, and S. xylosus. Enterotoxin genes were not identified in any CNS isolates, however sec and see were identified only in S. aureus associated with mastitis in goat. In compared to the isolates from 2006 to 2007, 27 S. aureus isolates from 2008 were found to be more resistant to ampicillin, cephalothin, oxacillin, oxytetracycline, penicillin G, and tetracycline. Eleven MRSA isolates were identified and belonged to SCCmec type III (nine isolates) as the major type and SCCmec type II (two isolates). These MRSA isolates revealed pulse-field gel electrophoresis (PFGE) pattern A (five isolates), C (one isolate), and D (one isolate) of human isolates. The other two isolates without pulsotypes belonged to ST59.

Conclusion

The prevalence and infection sites of CNS differed from those of CPS. Genetic analyses indicated that genetic divergence, possible zoonotic transfer of MRSA, and the involvement of sec as important virulence factors for of S. aureus that lead to mastitis in goats.

Two-dimensional immunoelectrophoresis was employed to electrophoretically identify membrane and cytoplasmic antigens of Mycoplasma arginini G-230. Five distinct cytoplasmic antigens were observed in soluble fractions prepared by digitonin lysis with electrophoretic mobilities (relative to bovine albumin) ranging from 0.36 to 0.86; four of these were common to other M. arginini strains: leonis and 23243. Five membrane antigens were identified, two of which (0.4 and 0.2) were common to the other M. arginini strains. The most prominent antigenic component of the membrane fraction (the complex membrane antigen) was electrophoretically heterogeneous, showing four antigenically related components with electrophoretic mobilities of 1.2, 0.95 to 0.76 and 0.05. The complex membrane antigen was exposed on the outside of the mycoplasmic cell because absorption of antiserum with live organisms removed antibody to this component. Antibodies to two other membrane components (0.6 and 0.2) were removed by absorption with Triton-solubilized membranes, but not by untreated membranes, indicating that these components were, at best, little exposed on either membrane surface. Antiserum was prepared against the complex membrane antigen using precipitin lines from two dimensional electropherograms as the immunogen. This antiserum reacted only with the complex membrane antigen and did not react with the other M. arginini strains, indicating that the complex membrane antigen was unique to strain G-230.

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The protective effect of sera produced in swine and goats exposed to virulent transmissible gastroenteritis virus (TGEV) or modified-live TGEV was tested in hysterectomy-derived, colostrum-deprived three-day-old pigs. Pigs were given serum with their daily ration of milk, and their immunity to virulent TGEV was determined. The pigs were observed for ten days for clinical signs of TGEV infection. One of nine pigs receiving goat serum was protected whereas all three pigs receiving three doses of swine serum per day were protected. Because virus was not isolated from the goats after oral/intranasal vaccination, it is suggested the virus did not replicate in either the respiratory or digestive tract of the goat.