Mycoplasmas and acholeplasmas isolated from ducks and their possible association with pasteurellas

Two hundred and sixty-three cases of clinically diseased ducks of all ages were examined for the presence of mycoplasmas. Mycoplasmas and acholeplasmas belonging to more than eight serogroups were cultured from 68 of them, and comprised 12 M anatis, one M columbinasale, two M gallinaceum, two M gallinarum, nine M synoviae, three unidentified Mycoplasma species, 37 Acholeplasma laidlawii and one unclassified acholeplasma belonging to each of serogroups 7 and 8. They were identified by biochemical characterisation, disc growth inhibition and agar gel diffusion tests. Fifty-three (78 per cent) of the isolates occurred with species of Pasteurella: 33.8 per cent with Pasteurella anatipestifer, 32.4 per cent with P multocida and 11.8 per cent with both P anatipestifer and P multocida. Nine of the isolates (13.2 per cent) were in pure culture and six (8.8 per cent) with other agents. Of the ducks negative for mycoplasmas 33.3 per cent were infected with P anatipestifer, 25.1 per cent with P multocida and 14.4 per cent with both P anatipestifer and P multocida. There was no correlation between the infections with mycoplasmas and P anatipestifer but there was a weak association between the infections with mycoplasmas, especially M anatis and P multocida.     

Detection certainty of the contamination of bull sperm with mycoplasma

Tests for presence of mycoplasmas were conducted on 20 insemination bulls known as mycoplasma spreaders, with 5 sperm pellet batches of 20 pellets each being investigated for each animal. Mycoplasma contamination was positively recorded from 83 of the above 100 batches. Mycoplasma (M.) bovigenitalium, M. californicum. M. arginini, M. bovirhinis, and Acholeplasma laidlawii were typed by means of indirect immunofluorescent technique, which confirmed the presence also in the GDR of mycoplasma species described in the literature and detected in sperm samples. The solid culturing media used in the above tests, medium-B agar and cattle blood agar with staphylococcal nutrix, proved to be equally suitable for isolation of mycoplasma from sperm samples. Mycoplasma was positively identified in about one third of all pellets/batches tested. 3 pellets to one batch should be sufficient a random sample size from which to obtain information at least very close to real contamination.     

Increasing prevalence of Mycoplasma bovis in Danish cattle

A study on the prevalence of mycoplasmas in pneumonic bovine lungs was performed on material submitted for diagnostic purposes at the Danish Veterinary Laboratory, Copenhagen. Among the 50 examined cases 43 (86.0%) were found to be infected with mycoplasmas. The predominant mycoplasmas were Ureaplasma spp. (72.0%), M. dispar (48.0%) and M. bovis (24.0%). Other mycoplasmas were M. bovirhinis (20.0%) and M. bovigenitalium (6.0%). Among the infected lungs multiple species infections were predominant (76.7%) over single species infections (23.3%) with M. dispar-Ureaplasma (25.6%), M. bovis-Ureaplasma (18.6%) and M. dispar-M. bovirhinis-Ureaplasma (11.6%) infections being the most frequently encountered combinations. There appears to be an increasing prevalence of M. bovis (24.0%) as compared to earlier reports (0.6-2.0%), thus calling for special attention upon this mycoplasma. Pulsed field gel electrophoresis (PFGE) analysis of 11 field isolates of M. bovis from 9 different farms revealed different profiles except for 2 isolates which were recovered from the same farm. Because mycoplasmas belonging to the 'M. mycoides cluster' were not encountered during this study; it appears that the Danish cattle population is still free from this group of mycoplasma in spite of their presence in some other European countries.     

In vitro susceptibilities to fluoroquinolones in current and archived Mycoplasma gallisepticum and Mycoplasma synoviae isolates from meat-type turkeys

Monitoring of susceptibility to antibiotics in field isolates of pathogenic avian mycoplasmas is important for appropriate choice of treatment. Our study compared in vitro susceptibility to enrofloxacin and difloxacin in recent (2005-2006) isolates of Mycoplasma gallisepticum and Mycoplasma synoviae from meat-type turkey flocks with archived (1997-2003) isolates and reference strains. Comparison of minimal inhibitory concentration (MIC) values determined by microtest, agar dilution and commercial Etest showed good agreement, but underscored the need for standardized methods for testing. Notably, while the commercial Etest was convenient and accurate for determining MICs for enrofloxacin in the range 0.002-0.094mug/ml, the endpoint of inhibition for M. gallisepticum and M. synoviae strains with MIC values >/=1.0mug/ml could not be determined. A decrease in susceptibility to both fluoroquinolones was detected in archived strains but to a greater degree in recent isolates, most of which had MICs above the NCCLS susceptibility breakpoint for these antibiotics (相似文献   

Detection of mycoplasmas in eye swab specimens of cattle

In the present investigation 262 conjunctival swabs were taken from 178 cattle and examined for mycoplasmas. The isolation was possible from 111 swabs. Mycoplasmas were found in eyes with clinical symptoms of IBK (in 64 of 148 swabs investigated = 43.2%) as well as in healthy eyes (in 47 of 114 swabs investigated = 41.2%). Consequently a correlation between clinical findings and isolation of mycoplasmas could not be observed. Unfortunately 60 of 111 isolates could not be subcultivated after storage at -20 degrees C. Using the indirect immunofluorescence test 41 of the 42 surviving isolates were identified as M. bovoculi which before has not been isolated in the Federal Republic of Germany. One isolate was determined as A. laidlawii. The 17 M. bovoculi strains investigated for their biochemical reactions showed the same characteristics like the reference strain M. bovoculi M 165/69. In repeating examinations mycoplasmas could be isolated 5 times after one month and 14 times after 6 months. Cattle younger than 2 years were more often infected with mycoplasmas (62.5%) than older animals (19.4%). No difference, however, could be observed in the clinical manifestations of IBK between younger and older animals. Mycoplasmas were more frequently isolated in autumn (43.6%) than in spring (21.4%) and summer (29.3%). In most of the animals examined both eyes were colonized by mycoplasmas. No spiroplasmas could be detected in the 262 conjunctival swabs investigated.     

我国鸭疫里氏杆菌血清型的鉴定

１９９７年１月－１９９８年３月,从北京市２０个商品鸭场自然病死的北京白鸭和河南省与上海市部分鸭场的樱桃谷鸭分离到２７６株鸭疫里氏杆菌,采用凝集试验和琼脂扩散沉淀试验,对其进行了血清型的研究。其中７０株细菌为１型,６４株为２型,其余１４２株怀１,２,型参考菌株的抗血清发生反应。     

Application of polymerase chain reaction fingerprinting to differentiate Ornithobacterium rhinotracheale isolates.

Ornithobacterium rhinotracheale (ORT) is an infectious respiratory pathogen of chickens, turkeys, and wild birds. There are 18 serotypes of ORT reported worldwide. In this study, enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction and random amplified polymorphic DNA assay with Universal M13 primer-based fingerprinting techniques were investigated for their ability to differentiate ORT isolates. The authors examined 50 field isolates and 8 reference strains of ORT for their genetic differences. The fingerprint patterns were compared with serotyping results of ORT by the agar gel precipitation test. M13 fingerprinting revealed different patterns for 6 reference serotypes of ORT that were tested, namely, C, D, E, I, J, and K. Ornithobacterium rhinotracheale reference serotypes A and F yielded indistinguishable fingerprints with M13 fingerprinting. The ERIC 1R technique discerned only 5 of the 8 reference serotypes of ORT. Distinct fingerprints were also found within the ORT serotypes with both techniques. From 58 isolates of ORT that were fingerprinted belonging to 8 ORT serotypes, 10 different fingerprints were obtained with M13 fingerprinting and 6 different fingerprints were obtained with ERIC 1R fingerprinting. M13 fingerprinting technique was found to be more discriminative in differentiating ORT isolates than the ERIC 1R fingerprinting technique. These results suggest that fingerprinting techniques may be a more discerning tool for characterizing ORT isolates than the serological test using the agar gel precipitation test. This fingerprinting technique could potentially be a valuable tool in identifying an isolate from a clinical outbreak of ORT infection for development of an autogenous vaccine.     

African Swine Fever: Application of Immunoelectroosmophoresis for the Detection of Antibody

Thirty-three pigs in three groups of nineteen, ten, and four pigs were infected with three different African swine fever (ASF) virus isolates, respectively. All virus isolates were attenuated to varying degrees by passaging in cell cultures, and they retained sufficiently low virulence to produce subacute and chronic infections in pigs. Sera collected at various intervals were tested for antibody activity by the immunoelectroosmophoresis, agar gel diffusion precipitin, and complement-fixation tests using a modified Kolmer technique. Results clearly indicated that the immunoelectroosmophoresis test is a rapid (30 minute) and accurate method with extreme sensitivity and superior to the complement-fixation and agar gel diffusion precipitin tests in detecting antibody against ASF virus. Possible use of this method in detecting ASF virus infection is suggested.     

Urinary tract infections due to Mycoplasma canis in dogs

Urine samples were obtained from 100 dogs with symptoms of lower urinary tract disease by cystocentesis and were examined for mycoplasmas. Urinalysis, haematological and biochemical analyses were also performed. Bacteria were isolated from urine in 41 of 100 dogs; Mycoplasma canis was isolated from four of 100 (4%) urine samples and three were pure culture. Selective mycoplasma media were used for isolation. In growth inhibition test, propagation of the four M. canis isolates was inhibited by their specific hyperimmune sera and there was no cross reactivity between isolates and hyperimmune sera of other mycoplasmas. Dogs in which M. canis was isolated were azotemic. All dogs were treated with enrofloxacin, furosemide, and supportive therapy (fluid therapy, ascorbic acid). In all animals, clinical improvements were observed after treatment.     

Evaluation of a digitonin disk assay to discriminate between acholeplasma and mycoplasma isolates from bovine milk

A study was undertaken to evaluate effectiveness of a digitonin disk inhibition test to discriminate between Acholeplasma laidlawii and Mycoplasma sp. isolated from bovine milk. The test measured zone diameters of growth inhibition surrounding a digitonin-containing disk on solid medium. Zones of inhibition for 20 isolates of A. laidlawii, ranging from 8-14 mm, did not overlap those of 261 isolates of Mycoplasma sp., ranging from 16 to 38 mm. Examination of variation in zone diameters for M. bovis found that inhibition was not appreciably affected by agar dehydration. Zones of inhibition increased with increasing dilutions of stock culture and decreased with increasing incubation time. Analysis of variance and Fisher's least significant difference test of logn zone diameters revealed that differences in mean logn zone diameters were different at the 0.01 level of significance between some of the six species of mycoplasma examined, indicating that growth among some species of mycoplasma was effected differently by digitonin. The digitonin test was found to clearly discriminate between A. laidlawii and Mycoplasma sp. indicating that the test would be useful as a practical screening test of individual-cow and bulk tank milk for mycoplasmas.     

ECOLOGY OF MYCOPLASMAS IN CLINICALLY HEALTHY CATS

Mycoplasmas were isolated from various sites and organs of a series of 319 clinically healthy cats. They include M. felis, M. gateae, M. Arginini, A. laidlawii, feline ureaplasmas, M. pulmonis, M. arthritidis, and M. gallisepticum. In addition, there were 10 strains of mycoplasmas which could not be identified with the specific antiserums by growth inhibition tests. Antibody against M. felis was demonstrated by haemagglutination-inhibition and complement-fixation tests in cats which were over 6 months of age. However, no such antibody against M. felis was detected in animals which were less than 6 months old. No antibody against A. laidlawii, M. gateae, M. arginini and feline ureaplasmas was demonstrated by the same serological methods. The significance of these mycoplasmas in cats is described.     

Evaluation of growth of avian mycoplasmas on bile salt agar and in bile broth

All nine Mycoplasma iowae strains and one strain of M gallinarum grew on 0.05 per cent 'bile' agar medium. The colony size of M iowae on this agar medium was similar to the size obtained on bile-free mycoplasma agar. One strain each of M maculosum, M arginini and M bovis also grew on 0.05 per cent bile agar. However, one strain each of M gallisepticum and M meleagridis were inhibited at this concentration. Six of the nine strains of M iowae were also resistant to 1 per cent 'bile' in broth medium but all were resistant to 0.5 per cent. The resistance of M iowae to 0.5 per cent 'bile' in broth may be a useful characteristic for differentiating it from some of the other avian mycoplasmas.     

鸭源禽流感病毒的分离和鉴定

从南京鸡鸭加工厂、孝陵卫鸭场及迈皋桥鸭场共采制１１３个泄殖腔拭子,用鸡胚尿囊腔接种传代法分离到４株禽流感病毒（ＡＩＶ）,每个样品均有ＮＤＶ混感。纯化后的分离株利用电镜负染技术观察到典型的ＡＩＶ粒子。琼扩试验证明４个分离株均为Ａ型流感病毒。经血凝素亚型分析,４株均属于Ｈ９亚型。致病性试验结果表明,４株鸭源禽流感病毒对鸡均表现为较低的致病力。本研究提示环境（特别是水体）保毒是ＡＩＶ得以长期存在和传播的重要因素和媒介。     

血清10型鸭疫里默氏菌第5个血清亚型的分析

采用琼脂扩散沉淀试验、玻片和试管凝集试验以及血清吸收凝集试验，对6株鸭疫里默氏菌分离株进行了抗原性分析。这6株分离株被鉴定为血清10型，但它们与10型内已知的4个亚型菌株之间又存在明显的抗原差异，因此，以菌株C919为代表的6个分离株被鉴定为血清10型的第5个亚型。     

Phenotypic and genotypic characteristics of Mycobacterium isolates from fighting fish Betta spp. in Malaysia

Mycobacteriosis due to mycobacteria is one of the most common bacterial diseases in ornamental fish. We describe here the phenotypic and genotypic characteristics of Mycobacterium isolates from fighting fish Betta spp. using ATCC Mycobacterium marinum, Mycobacterium fortuitum and Mycobacterium chelonae as references. A total of four isolates (M1, M2, M3, M4) were obtained from four out of 106 fish samples using selective agar, and identified to Mycobacterium genus using acid-fast staining and 16s rRNA gene-based genus specific polymerase chain reaction. DNA sequencing and NCBI-BLAST analysis further identified isolate M1 as M. marinum and isolates M2, M3, M4 as M. fortuitum. Morphological, physiological and biochemical tests were carried out for phenotypic characterizations. Universal M13 and wild-type phage M13 RAPD dendogram was generated to illustrate the genetic relationship of the isolates and reference strains.     

Investigations on different Ornithobacterium rhinotracheale "ORT" isolates

The aim of the present investigation was to determine the antigenic relationship between different Ornithobacterium rhinotracheale (ORT) isolates and to serotype field isolates obtained from turkey and chickens. Different antigen extractions (heat-stable, proteinase K-stable [lipopolysaccharide], and sodium dodecyl sulfate [SDS] extractions) were prepared from each serotype (A, B, C, D, E, and G) as well as from 21 ORT field isolates and examined in agar gel precipitation (AGP) and enzyme-linked immunosorbent assay (ELISA) tests. The field isolates were cultured from turkey (16 isolates) and chicken (5 isolates) flocks showing respiratory manifestations. Monospecific reactions were obtained with heat-stable as well as proteinase K-stable antigens prepared from serotypes A, C, D, E, and G in AGP tests. On the other hand, with the same antigen preparations from a strain of serotype B in AGP tests, cross-reactions with antisera prepared against serotypes A and E could be detected. The cross-reactions were observed mostly between 48 and 72 hr. In applications of SDS-antigen preparations in AGP tests, cross-reactions between all serotypes except serotype C were detected between 24 and 72 hr. Testing all antigen preparation in ELISA, different cross-reactions were observed and the evaluation of the results is very difficult. Serotyping of the field isolates in AGP tests by using heat-extracted antigens showed after 24 hr that 10 out of 16 isolates from turkey belonged to serotype B, five to serotype A, and one to serotype E. Results obtained after 48-72 hr revealed cross-reactions between serotype B and E in 11 cases and between A and B in two cases. All five isolates obtained from chicken reacted after 24 hr only with serum against serotype A. After 48-72 hr, two isolates showed cross-reaction with antiserum against serotype B. Similar results were obtained with proteinase K-stable antigen.     

Isolation and identification of mycoplasmas from the nasal cavity of sheep

Mycoplasmas isolated from the nasal cavity of sheep in a ram test station were examined to determine their identity and prevalence. Specimens were obtained for mycoplasmal culture in 1980, 1982, and 1983 from 558 sheep, and mycoplasmas were isolated from 630 specimens from 320 sheep (57.3%). The isolates were characterized and differentiated into groups on the basis of sensitivity to digitonin, fermentation of glucose, and hydrolysis of arginine. Isolates in some groups were further characterized by use of additional diagnostic media, and their identity was confirmed by agglutination or growth inhibition with antiserum prepared from reference mycoplasmas. Of the 320 sheep with mycoplasmas, 293 had Mycoplasma ovipneumoniae, 12 had M arginini, and 1 had M capricolum. Two sheep had Acholeplasma spp, and 3 sheep had unidentified Mycoplasma spp. The remaining 9 sheep had M ovipneumoniae in combination with Acholeplasma spp (n = 3), M arginini (n = 3), M capricolum (n = 2), and an unidentified Mycoplasma spp (n = 1). The biochemical reactions of the M ovipneumoniae from the 293 sheep were similar, but varied in the degree of growth and fermentation in the basal medium containing glucose. The high prevalence of M ovipneumoniae indicated that it may be commensal in the upper respiratory tract of healthy sheep.     

Characteristics of Actinomyces pyogenes involved in lameness of male turkeys in north-central United States

A new condition of clinical lameness in 20 male turkey flocks of North-Central United States, associated with isolation of gram-positive rod bacteria from lesions of osteomyelitis, is characterized. The characterization confirmed the randomly selected isolates as Actinomyces pyogenes based on macroscopic and microscopic observations and 17 biochemical tests. The disease was reproduced within 3 weeks in all male turkeys, following an intravenous challenge at 15 weeks of age. The agar gel precipitin test and immunoblotting confirmed the antigenic similarity of the isolates recovered from the osteomyelitis lesions of lame birds.     

The occurrence of mycoplasmas in the lungs of pigs in New Zealand

Attempts were made to recover and serologically identify mycoplasmas from the lungs of 50 pigs with mycoplasmal pneumonia and from 50 lungs without gross evidence of pneumonia. Mycoplasma hyopneumoniae and M. hyorhinis were respectively cultured from 30% and 50% of pneumonic lungs and the former species was also recovered from 12% of non-pneumonic lungs. Three other isolates (one from pneumonic and two from non-pneumonic lungs) differed in colonial morphology from M. hyopneumoniae and M. hyorhinis. The viability of these isolates could not be maintained on subculture and they were not identified serologically. The indirect immunofluorescence test was found to be highly specific for the identification and differentiation of M. hyopneumoniae and M. hyorhinis.