Serological reactivity to Mycobacterium bovis protein antigens in cattle.

The serological response to 12 purified Mycobacterium bovis antigens were examined in an ELISA assay. These antigens included the majority of M. bovis protein antigens described to date and in most cases they were very similar to the M. tuberculosis antigens of the same molecular mass.

The purified antigens were tested against sera from M. bovis infected cattle, M. bovis culture-negative cattle from infected herds and animals infected with related microorganisms, mainly other mycobacterial species. All the antigens gave strong reactions with at least some sera from the M. bovis infected group and showed cross-reactivity with some of the sera from the other two groups. The antigen with the highest specificity reacted strongly with only 60% of the M. bovis infected sera. Antigens that reacted with most or all of the M. bovis infected sera also gave the highest cross-reactivity with sera from the other two groups. These results indicate that a serological test based on any one or a combination of these antigens, without removal of the cross-reacting epitopes, would be unsatisfactory.     

Mapping of IS6110 insertion sites in Mycobacterium bovis isolates in relation to adaptation from the animal to human host

The physiological role and impact of IS6110 insertions on the biology of Mycobacterium tuberculosis complex is not well understood. Insertion of IS6110 in coding regions can cause loss of gene activity, while homologous recombination between two copies of IS6110 can result in the deletion of genes or in rearrangement of genomic regions involved. In addition to these genomic changes, IS6110 can also activate flanking genes through acting as a mobile promoter.

In order to determine the possible role of IS6110 transposition in the adaptation to humans, we selected Mycobacterium bovis isolates from endogenous reactivation cases in elderly people in The Netherlands. The human isolates contained higher number of IS6110 copies in comparison to the bovine M. bovis strains. These additional integration sites of IS6110 were sequenced and analyzed. From 12 of such IS6110 insertion sites, 6 loci were located in the intergenic regions, and 6 other occurred within coding regions. IS6110 was inserted in a position where it might serve as a promoter in two cases. We conclude that IS6110 transpositions in M. bovis may be a driving force in the adaptation from the animal to the human host.     

犊牛支原体肺炎继发牛A型多杀性巴氏杆菌感染的诊治

某奶牛场犊牛相继发生肺炎和关节炎,为确诊该牛场犊牛群发病的原因并提出防控方案,本试验剖检新生犊牛并采集病料,分别开展牛支原体及其他病原菌的分离培养、PCR鉴定及药敏分析;进行牛病毒性腹泻病毒、牛口蹄疫病毒和牛传染性鼻气管炎病毒PCR检测;制作犊牛肺脏组织病理切片并进行观察和评估。从犊牛肺脏组织分离到牛支原体和牛A型多杀性巴氏杆菌;牛病毒性腹泻病毒、牛口蹄疫病毒和牛传染性鼻气管炎病毒检测均为阴性;肺脏组织病理切片可见肺泡结构破坏、出血及大量炎性细胞浸润;药敏试验结果显示,牛支原体和牛A型多杀性巴氏杆菌分别对泰乐菌素和头孢唑啉敏感,但对青霉素、庆大霉素、林可霉素和氨苄西林均呈现耐药。该犊牛群确诊为牛支原体肺炎继发牛A型多杀性巴氏杆菌感染,采用泰乐菌素联合头孢唑啉肌肉注射,配合对症治疗和规范管理,有效控制了该场犊牛疾病。     

新疆石河子地区牛支原体的分离鉴定及药物敏感性分析

【目的】确定引起新疆石河子地区集约化牛场常发性肺炎的主要病原同时进行病原的体外药物敏感性分析。【方法】采集有典型咳嗽、流涕症状的牛鼻拭子10份和病死牛肺脏组织1份,用牛支原体特异性引物进行PCR检测,将检测为阳性的样本进行病原培养纯化,对纯化后的分离株菌落进行形态学观察、Dienes染色、生化试验及16S rRNA测序和进化分析,通过测定颜色变化单位(CCU)测定分离株生长曲线,并对分离株进行药物敏感性试验。【结果】PCR结果显示,10份鼻拭子中检测出7份牛支原体阳性样本,1份病死牛肺脏组织也检测为阳性;在涂有肺脏组织研磨液培养液的PPLO固体培养基上长出针尖状的菌落,纯化后分离株菌落形态为典型的煎蛋状;Dienes染色可见明显的深蓝色中心脐;生化试验结果显示,分离株不水解明胶、精氨酸、七叶苷,不发酵乳糖、葡萄糖和甘露醇,不分解尿素,可还原氯化三苯基四氮唑;16S rRNA测序结果显示,分离株与牛支原体国际标准株PG45相似性为99.7%,与国内牛支原体地方流行株XBY01、Ningxia-1、NM2012、Tibet-10的相似性最高,均为99.9%;生长曲线测定结果显示,分离株在培...     

Analyses of lipopolysaccharides, outer membrane proteins and DNA fingerprints reveal intraspecies diversity in Moraxella bovis isolated in Argentina

Intra-specific diversity within Moraxella bovis was investigated analysing DNA fingerprints, outer membrane proteins (OMP) and lipopolysaccharides (LPS) profiles. Three collection strains and 57 isolates of M. bovis, collected during 3 years from cattle with infectious bovine keratoconjunctivitis (IBK) symptoms, from diverse geographical locations of Argentina, were examined. The LPS and OMP profiles were studied through SDS–PAGE analysis and genotype was determined by PCR-DNA fingerprinting. Genotyping identified five DNA types while analysis of LPS and OMP profiles identified three rough LPS types and three OMP types among the 60 isolates of M. bovis including the three collection strains. None of the three methods employed to assess diversity was discriminating when used alone because the degree of heterogeneity in each group of surface structures was limited, but when data of each typing method were combined, 15 distinct subgroups were determined. This subgrouping was clearly able to differentiate isolates of the same genotype. These typing methods appear to be useful to assess different aspects of the disease such as the diversity within a population of M. bovis associated to epidemic conditions, track the causal agent in an outbreak of the disease, monitoring vaccination programs and studies on virulence.     

Protein variability among Mycoplasma hyopneumoniae isolates

Sodium-dodecyl-sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was used to study the protein variability of Mycoplasma hyopneumoniae isolates. Fifty-six M. hyopneumoniae isolates from 6 different countries and 37 different herds were used. From eight herds, more than one isolate was available. All SDS-PAGE patterns of isolates originating from different herds were clearly divergent. Intra-species protein variability was quantified using the reference strain J and seven field strains all obtained from different herds and classified according to virulence. Between the field strains, a variability of 25% was found, while the culture-adapted strain J was clearly divergent and showed 30% variability with the field strains. No clustering according to virulence was obtained, but a protein band of about 181 kDa was present in the two highly virulent isolates whereas this protein band was absent in the moderately and low virulent isolates. Protein patterns of isolates derived from different animals from the same herd, were identical or differed in only a few protein bands. This study clearly indicates that, in agreement with previous studies on genomic diversity of M. hyopneumoniae isolates, proteomic variability within the species is high. Our study did not find clear evidence that more than one M. hyopneumoniae isolate circulates within a herd at a specific time point. The minor differences found between M. hyopneumoniae isolates from the same herd might reflect the organism's ability to alter its proteomic expression profile under field conditions.     

分枝杆菌分型三重PCR检测方法的建立及应用

本研究旨在建立一种快速鉴定分枝杆菌的三重PCR方法,并比较分析其在临床检测中的可靠性。根据已发表的结核分枝杆菌、牛分枝杆菌和非洲分枝杆菌rv 3036c基因,结核分枝杆菌rv 1970f基因(RD7)和牛分枝杆菌pncA基因的序列,改造并设计合成了3对特异性扩增引物,建立了一种能对分枝杆菌样品进行初步鉴定的三重PCR方法。结果显示该方法可针对rv 3036c、rv 1970f和pncA基因分别扩增出大小为500、125和249 bp的目的片段,能特异性检测出结核分枝杆菌(500和125 bp两条带)和牛分枝杆菌(500和249 bp两条带),并可将结核分枝杆菌、牛分枝杆菌与其他分枝杆菌加以区分。本方法的检测灵敏度为50 pg/μL模板基因组DNA。对86株抗酸染色阳性菌进行三重PCR鉴定,鉴定结果与细菌16S rDNA和ITS序列测定结果一致,检测准确度为100%,优于生长特征和生化试验鉴定。     

Genomic lineage of Salmonella enterica serovar Dublin

Thirty five strains of the host adapted Salmonella serotype Dublin (S. Dublin) have been characterized by IS200 patterns, ribotyping, pulsed-field gel electrophoresis (PFGE), restriction fragment polymorphism after hybridization with five randomly cloned DNA-framents of S. enteridis (RFLP), and plasmid profiling in order to divide the strains into ‘genomic lines’. For comparison, 20 other strains of 9 different group-D serotypes were included. The IS200 patterns were identical in all strains of S. Dublin examined. These patterns were different from those observed in other group-D Salmonella with the exception of one strain S. Enteritidis phage type 11 and a strain of S. Rostock. The insertion element IS200 was not detected in strains of S. Dar-es-Salam, S. (II) 9,12:z -, and S. Panama. RFLP, based on probing with five random cloned chromosomal fragments gave the same pattern in all strains except for one isolate from the UK. This strain was also found to have an unique PFGE pattern and ribotype. Among the remaining strains, three different PFGE patterns and 7 different ribotypes were observed. Based on all four typing methods, 8 different ‘genomic lines’ of S. Dublin were identified. The same grouping could be obtained from the use of ribotyping alone, but PFGE and RFLP were found to provide valuable information on possible relationships between ribotypes. Seven different plasmid profiles and a group of strains without plasmids were observed. In several cases, the same plasmid profile was shown to be present in more than one ‘genomic line’.     

Establishment and Application of Mycobacterium Typing Triple PCR Detection Method

This study was aimed to establish a triple PCR method to rapidly identify Mycobacterium species, and evaluate its testing reliability.Three pairs of primer that were respectively specific to rv 3036c, rv 1970f and pncA genes of Mycobacterium were designed to establish a triple PCR for preliminary identification of Mycobacterium tuberculosis(M.tuberculosis), Mycobacterium bovis(M.bovis) and other Mycobacterium spp.PCR products were the expected sizes of 500(rv3036c), 125(rv1970f) and 249 bp(pncA), and contained two DNA bands(500 and 125 bp) with M.tuberculosis DNA template, two DNA bands(500 and 249 bp) with M.bovis DNA template.No band or non-specific band appeared with Mycobacterium spp.except M.tuberculosis and M.bovis DNA templates.The sensitivity of the triple PCR was calculated to 50 pg/μL template of genomic DNA.86 acid-fast bacteria were detected by the triple PCR, 16S rDNA and ITS gene sequencing, growth test and biochemical test, and the results were consistent between triple PCR and 16S rDNA and ITS gene sequencing.The detecting accuracy of triple PCR was 100%, and higher than growth test and biochemical test.     

Identification of an immunodominant 40 kDa merozoite antigen common to the Australian T and Dixie vaccine strains of Babesia bovis and the development of diagnostic tests specific for these strains

Antigenic differences among Australian vaccine and field strains of Babesia bovis were investigated in an attempt to identify strain specific antigens. Immunoblots revealed substantial differences between the current vaccine strains, designated T and Dixie, and previous vaccine strains and field isolates collected on properties where vaccination with the T or Dixie strains had failed to provide complete protection against tick-borne challenge. A major difference was an immunodominant 40 kDa antigen (T40) present in only the T and Dixie strains. The molecular weight and immunodominant nature of this antigen suggest that it may be the equivalent of the major merozoite surface antigen (MSA-1) described by others in North American strains of B. bovis. MSA-1 was shown to be conserved in north American isolates but not in an isolate from Israel or in the Australian S and L isolates. The work presented here suggests that merozoite surface antigen diversity exists among geographically different isolates of B. bovis within Australia.

Monospecific antiserum to T40 was used to develop an indirect fluorescent antibody (IFA) test specific for T and Dixie strain parasites, and a blocking enzyme-linked immunosorbent assay (ELISA) specific for antibody to the T and Dixie strains. In cases of babesiosis in recently vaccinated cattle, the IFA test will be a useful tool for determining whether clinical symptoms are due to a severe vaccine reaction or to a concurrent tick-borne infection. In a preliminary assessment of potential of the ELISA for the serological identification of vaccinated cattle using a total of 160 sera, the test clearly differentiated between animals vaccinated with the T or Dixie strains and non-vaccinated animals, and was not affected by presence of antibodies to other B. bovis strains.     

Tuberculosis as a zoonosis from a veterinary perspective

Tuberculosis is an important disease among many zoonoses, because both Mycobacterium tuberculosis and Mycobacterium bovis, which are the major causes of tuberculosis, are highly pathogenic, infect many animal species and thus are likely to be the source of infection in humans. In particular, monkeys are highly susceptible to these bacteria and are important spreaders. Recently, two outbreaks of M. tuberculosis occurred in four different kinds of monkeys and humans were also infected with the disease in Japan. In zoos, tuberculosis was reported not only in monkeys, but also in several different kinds of animals, including elephants. Pets such as dogs and cats are believed to be generally less susceptible to M. tuberculosis, but in this article we introduce a case of infection from man to dog by close contact. Japan is one of the few countries that have been able to control M. bovis infection. In other countries, however, cases of bovine tuberculosis and human M. bovis infection have been reported, and thus further attention is still required in the future.     

Immunoblot analysis of cyanogen bromide-cleaved Moraxella bovis pilin reveals presence of shared antigenic determinants on pili from heterologous strains

Moraxella bovis pilus proteins, collected and purified from four strains of M. bovis, were cleaved with cyanogen bromide. Two major fragments were produced. Antisera were produced in rabbits to the pilin protein fragments and to whole uncleaved pili from these strains. Immunoblots of whole and cyanogen bromide-cleaved pilin were reacted with the homologous and heterologous antisera to whole pili and cleaved pilin. Antisera to whole pili reacted strongly with homologous pilin. Weaker and inconsistent reactions were detected with heterologous pilin. Antisera produced to cyanogen bromide-cleaved pilin proteins reacted strongly with homologous and heterologous pilin fragments and uncleaved pilin proteins. These findings demonstrate the presence of conserved antigenic determinants on pili from heterologous strains that are non-immunogenic in the intact pilus but are immunogenic after treatment with cyanogen bromide. Cyanogen bromide-treated pilus preparation might have potential as a vaccine because antibodies are induced against heterologous strains of M. bovis, whether these cross-reactive antibodies are protective remains to be determined.     

Isolation of mycoplasmas from milk, lungs and prepuce of cattle

M. bovis was mainly isolated from 6.25% of milk samples collected from cows with clinical and sub-clinical mastitis. The main mycoplasma strain isolated from the respiratory tract of calves with symptoms of bronchopneumonia was M. bovirhinis (12.1% of samples). M. bovigenitalium was most frequently recovered from bull's prepuce (67.1% of samples).     

Genotyping of Mycobacterium bovis by geographic location within Mexico

The spacer oligonucleotide typing (spoligotyping) method was used to differentiate 62 Mycobacterium bovis isolates obtained from tissues with macroscopic lesions typical of tuberculosis in dairy cattle from different regions of Mexico. Our purpose was to see if a strain from one region was genetically different from those of other regions (with the long-term aim of doing molecular trace back of isolates obtained in the laboratory). Results from the genetic analysis indicate that M. bovis isolates cannot be grouped by geographic location due to a wide range of genetic types involved in dairy cattle infections. Isolates even from the same herd showed different spoligotypes but some isolates from different region had similar genetic patterns. Genetic typing without epidemiologic information does not seem to be a plausible method to trace back animals to source of origin to detect and eliminate sources of infection.     

铝和酸胁迫对苜蓿根瘤菌生长和抗氧化酶系的影响

在铝和酸胁迫下,比较分析了天蓝苜蓿和紫花苜蓿根瘤菌的生长及SOD(超氧化物歧化酶)、CAT(过氧化氢酶)、POD(过氧化物酶)、及GR(谷胱甘肽还原酶) 等抗氧化酶系响应特点。铝胁迫设5个梯度,即0,25,50,75,100 μmol/L;酸胁迫设4个pH水平,即4,5,6,7。结果表明,铝和酸胁迫下,2种苜蓿根瘤菌的OD_A600均显著降低,生长受到抑制,且天蓝苜蓿根瘤菌的OD_A600显著高于紫花苜蓿根瘤菌。在pH水平为4和5时,紫花苜蓿根瘤菌没有生长。在铝毒胁迫下,天蓝苜蓿根瘤菌的SOD酶活性一定程度减小后缓慢上升,而CAT、POD和GR酶活性随铝浓度增大显著下降;紫花苜蓿根瘤菌的SOD,CAT及GR酶活在低Al胁迫下无显著变化,在高铝胁迫下显著下降。随着pH水平的下降,天蓝苜蓿和紫花苜蓿根瘤菌均表现为SOD、CAT、POD、GR酶活性的显著下降。在不同铝及酸胁迫下,天蓝苜蓿根瘤菌各抗氧化酶活性显著高于紫花苜蓿根瘤菌。综合分析认为天蓝苜蓿根瘤菌较之紫花苜蓿根瘤菌有较强的耐酸、耐铝性。     

青贮剂对再生稻头季全株青贮品质和体外瘤胃发酵特性的影响

旨在探究青贮剂对再生稻头季全株青贮品质和体外瘤胃发酵特性的影响。以再生稻头季全株为青贮原料,在青贮中分别添加植物乳杆菌（60%）+纤维素酶（30%）+木聚糖酶（10%）、植物乳杆菌（70%）+粪肠球菌（20%）+纤维素酶（5%）+半纤维素酶（5%）、植物乳杆菌（30%）+粪肠球菌（60%）+纤维素酶（5%）+半纤维素酶（5%）、等量水为对照（依次记为N₁、N₂、N₃和CK组,各组添加量为500 g·t^-1 鲜重）。青贮45 d后开包取样,分析其青贮品质和体外瘤胃发酵特性。结果表明：1）青贮结束后,N₁、N₂和N₃组的总能显著高于CK组（P<0.05）,中性洗涤纤维和酸性洗涤纤维含量显著低于CK组（P<0.05）,且N₂组的中性洗涤纤维含量显著低于N₁和N₃组（P<0.05）,N₁、N₂和N₃组的总可消化养分含量和乙酸含量均显著高于CK组（P<0.05）,氨态氮/总氮显著低于CK组（P<0.05）。2）体外发酵24 h后,N₂组的24 h累积产气量和快速降解部分的产气量显著高于N₁、N₃和CK组（P<0.05）,N₂组的潜在产气量显著高于N₁、N₃和CK组（P<0.05）,且N₁组显著高于CK组（P<0.05）,N₁和N₂组的pH显著低于N₃和CK组（P<0.05）,N₂组的乙酸和氨态氮含量显著高于N₁、N₃和CK组（P<0.05）,N₁和N₂组的丙酸含量显著高于N₃和CK组（P<0.05）,N₂组乙酸/丙酸显著低于N₃和CK组（P<0.05）,N₂和N₃组的干物质降解率和粗蛋白质降解率显著高于N₁和CK组（P<0.05）,N₂组的有机物降解率、中性洗涤纤维降解率和酸性洗涤纤维降解率显著高于N₁、N₃和CK组（P<0.05）。综上所述,在实际生产中,建议青贮剂在以植物乳杆菌为主要添加剂的前提下,辅助添加粪肠球菌、纤维素酶和半纤维素酶,有利于获得营养价值较佳的再生稻头季全株青贮。     

草地早熟禾叶片表皮特征、解剖结构及光合特性对不同施氮量的响应

为探究草地早熟禾叶片表皮特征、解剖结构及光合特性对施氮量的响应特性，本试验以6份草地早熟禾为材料，分别为3份美国引进品种：Merit、Jackrabbit和Park,2份山西本土野生居群：应县和浑源，1份由山西野生居群选育的‘太行草地早熟禾’品系，按照两个施氮水平N₁:10 g·m^-2(低氮)和N₂:40 g·m^-2(正常)进行处理，采用徒手切片法和石蜡切片法制片，光学显微镜下拍照并测定叶片上下表皮厚度、细胞长宽比、细胞密度、气孔指数、气孔长宽比、气孔密度，叶片厚度、厚壁组织厚度、维管束面积以及气孔导度和净光合速率等指标。结果表明：N₁处理下，草地早熟禾叶片的上下表皮厚度、叶片厚度、厚壁组织厚度、维管束面积、气孔导度和净光合速率显著低于N₂处理，而气孔大小与N₂处理无显著差异；N₂处理下，Jackrabbit和Park的叶片厚度显著高于浑源和应县，其中Park的叶片厚度最大，为173.39μm,Jackrabbi...     

Molecular fingerprinting confirms extensive cow-to-cow intra-herd transmission of a single Mycobacterium bovis strain

In this study we have characterized M. bovis isolates from a herd of cattle in Uvalde, Texas in which 52 of the 193 animals selected at random in 1994 from a herd of 331 were caudal fold skin-test positive. Thirty-two of 52 skin-test positive cattle had gross lesions at slaughter, and isolations of M. bovis were made from 29 animals. The herd was comprised of Red Devon cattle purchased between 1978 and 1980 (n = 26) and breeding bulls (n = 3) introduced at later times, and all were tuberculosis test negative at the time of purchase. Other animals were natural additions (offspring) of these cattle. One additional animal, a Holstein present on the ranch at the time of purchase in 1976, was retained to nurse orphaned and weak calves. Using several molecular fingerprinting techniques we have verified a clonal relationship among the M. bovis isolates consistent with infection originating with a single strain. The molecular fingerprint patterns demonstrate the stability of the profiles despite persistence and spread of the organism within the herd for two decades and confirms their use in epidemiological tracing.     

Colonisation of the respiratory tract of lambs by strains of Mycoplasma ovipneumoniae

The age and time of year when colonisation of the nasal cavity of lambs by Mycoplasma ovipneumoniae occurs; the persistence of the organism, and its prevalence in the lungs at slaughter were examined in 2 flocks of sheep in New Zealand. No colonisation had occurred at the time of weaning at 6–7 weeks, but M. ovipneumoniae was recovered from most lambs on at least one occasion before they were slaughtered when about 8 months old. In most cases, colonisation of the nasal cavity by M. ovipneumoniae was a transient phenomenon. At slaughter M. ovipneumoniae was recovered from the lungs of 89% of the lambs of one flock and 80% of the other flock.

Bacterial restriction endonuclease DNA analysis (BRENDA) of 34 nasal isolates from one flock showed that it was possible to identify 7 “groups” each with markedly different BRENDA patterns. Lambs initially colonised by one strain, often lost that strain, and if recolonisation occurred it was with a different strain.

M. ovipneumoniae was recovered at slaughter from the lungs of most lambs, both normal and pneumonic. The isolates from one flock were examined by BRENDA, and approximately 90% of them gave similar or identical patterns. The predominant strain isolated from the lungs had been recovered from the nasal cavity of many of the lambs about 3 weeks earlier. This suggests that the nasal and lung isolates do not represent independent populations. However, nasal strains may differ in their ability to colonise the lungs.     

Identification and localization of a 94 kDa membrane protein found in Mycoplasma bovoculi strains

Six isolates of Mycoplasma bovoculi obtained from cattle herds with bovine keratoconjunctivitis were analyzed by gel electrophoresis and immunoblotting techniques. All six strains showed similarity in their protein profiles although no two patterns were identical. Antigenic differences between strains were detected in immunoblots reacted with post-exposure calf serum. A common 94 kDa protein band designated p94 was detected in all six strains reacted with monoclonal antibody MA25.5 developed to one of the strains. The p94 was also recognized in these strains by the calf serum. Trypsin treatment of intact mycoplasma cells resulted in the removal of p94 from immunoblots reacted with MA or hyperimmune rabbit serum. Other trypsin-resistant antigens shared between strains or being strain-specific in nature were identified when trypsin-treated mycoplasma cells were reacted with hyperimmune rabbit serum. The p94 antigen was shown to be of mycoplasmal origin by radio-immunoprecipitation using the MA or hyperimmune rabbit serum. These studies identify the presence of a surface antigen (p94) on M. bovoculi membrane in all strains examined that is trypsin sensitive by the use of monoclonal antibody, calf serum and hyperimmune rabbit serum.

Résumé

Six souches de Mycoplasma bovoculi, d'origine bovine (animaux affectés de keratoconjunctivite) ont été analysées par des techniques d'éléctrophorèse et par ‘immunoblotting’. Les six souches ont montré des similitudes dans leurs profils proteiniques. Cependant, aucun profil n'a été identique. Des differences antigéniques entre les souches ont étés detectées suite au traitement des immunoblots avec un sérum bovin infecté par M. bovoculi. L'anticorps monoclonal MA25.5 dirigé contre une des souches a réagi avec une bande proteinique (94 kDa) commune aux six souches. Cette bande a été designee p94. Cette bande p94 a été également identifiée chez tous les souches en utilisant le sérum bovin. Le traitement des mycoplasmes intactes par la trypsine a éliminé la bande p94 des immunoblots: cela a été démontr é par l'usage d'un anticorps monoclonal, ou par un sérum hyperimmun de lapin. D'autres antigènes résistants à la trypsine, communs ou spécifiques aux souches ont été identifiés quand les mycoplasmes traités par le trypsine ont été mélangés au sérum hyperimmun de lapin. La radioimmunoprécipitation par l'anticorps monoclonal ou le sérum hyperimmun de lapin a demontré que l'antigène p94 est d'origine mycoplasmique. Ces études identifient la présence d'un antigène de surface (le p94) localisé sur la membranc de M. bovoculi chez toutes les souches examinées. Cet antigène est sensible à la trypsine. Cela a été montré avec l'anticorps monoclonal, le sérum bovin et le sérum hyperimmune du lapin.