Background Lymphocyte function-associated antigen-1 (LFA-1, CD11a/CD18, alphaLbeta2), the most abundant and widely expressed beta2-integrin, is required for many cellular adhesive interactions during the immune response. Many studies have shown that LFA-1 is centrally involved in the pathogenesis of several diseases caused by Repeats-in-toxin (RTX) -producing bacteria. Results The porcine-LFA-1 CD11a (alpha) subunit coding sequence was cloned, sequenced and compared with the available mammalian homologues in this study. Despite some focal differences, it shares all the main characteristics of these latter. Interestingly, as in sheep and humans, an allelic variant with a triplet insertion resulting in an additional Gln-744 was consistently identified, which suggests an allelic polymorphism that might be biologically relevant. Conclusion Together with the pig CD18-encoding cDNA, which has been available for a long time, the sequence data provided here will allow the successful expression of porcine CD11a, thus giving the first opportunity to express the Sus scrofa beta2-integrin LFA-1 in vitro as a tool to examine the specificities of inflammation in the porcine species.

A significant decrease in adherence rates of Mycoplasma bovis to bovine bronchial epithelial (BBE) cells has been observed after passage of the organism in artificial medium. Analysis of the proteins expressed by M. bovis isolate 2610 by two-dimensional (2-D) electrophoresis demonstrated differences between the cells harvested after the 7th and 116th passage. Three silver-stained prominent spots observed in 2-D electrophoretic separation of protein extracts of the lower-passaged cells were considerably less strongly expressed in the sample from higher-passaged cells. These spots had a molecular mass of approximately 24 kDa and an isoelectric point of about 5. The mass spectrometry analysis of these trypsin-sensitive proteins led to their identification as a unique new member of the Vsps family of membrane-associated proteins. Serum from a mouse immunized with these proteins significantly reduced adherence of M. bovis to BBE cells. This result underlines the function of this new Vsp in adherence of M. bovis to host cells.

Three new insertion elements, ISMbov1, ISMbov2 and ISMbov3, which are closely related to ISMag1 (Mycoplasma agalactiae), ISMmy1 and IS1634 (both Mycoplasma mycoides subsp. mycoides SC), respectively, have been discovered in Mycoplasma bovis, an important pathogen of cattle. Southern blotting showed that the genome of M. bovis harbours 6-12 copies of ISMbov1, 11-15 copies of ISMbov2 and 4-10 copies of ISMbov3, depending on the strain. A fourth insertion element, the IS30-like element, is present in 4-8 copies. This high number of IS elements in M. bovis, which represent a substantial part of its genome, and their relatedness with IS elements of both M. agalactiae and M. mycoides subsp. mycoides SC suggest the occurrence of two evolutionary events: (i) a divergent evolution into M. agalactiae and M. bovis upon infection of different hosts; (ii) a horizontal transfer of IS elements during co-infection with M. mycoides subsp. mycoides SC and M. bovis of a same bovine host.

There is scarce information about the frequency and epidemiological and clinical features associated with the presence of Mycoplasma spp. in Argentine dairy herds. The objectives of this study were to develop a multiplex PCR for identifying M. bovis and M. canadense and to describe the frequency of Mycoplasma spp. isolated from clinical samples submitted to a diagnostic laboratory. Of a total of 1548 samples from intramammary infections, bulk tank milk and biological fluids, 38 Mycoplasma isolates were obtained. M. bovis, M. canadense, M. californicum and M. leachii were detected by using two multiplex PCRs, confirming their presence in clinical conditions in dairy cattle. The techniques used in the present study can be useful to broaden the knowledge about Mycoplasma infections in cattle, since the search for these organisms is not usually included in routine diagnoses.Existe poca información sobre la frecuencia, así como las características epidemiológicas y clínicas asociadas con la presencia de Mycoplasma en los rodeos lecheros argentinos. Los objetivos de este estudio fueron desarrollar una PCR multiplex para identificar M. bovis y M. canadense y describir la frecuencia de especies de Mycoplasma aisladas de muestras clínicas enviadas a un laboratorio de diagnóstico. De un total de 1.548 muestras de infecciones intramamarias, leche de tanque de frío y fluidos biológicos, se obtuvieron 38 aislamientos de Mycoplasma. Mediante 2 PCR multiplex se detectaron M. bovis, M. canadense, M. californicum y M. leachii, confirmando su presencia en síndromes clínicos en ganado lechero. Las técnicas utilizadas en el presente estudio pueden ser útiles para ampliar el conocimiento sobre las infecciones por Mycoplasma en bovinos, ya que la búsqueda de estos organismos no suele incluirse en los diagnósticos de rutina.

Mycoplasma bovis is known to be responsible for pneumonia and arthritis in calves, as well as mastitis in dairy cows. Despite clear evidence of its pathogenic potential, little is known about mechanisms of cytadherence and the molecular factors involved. The purpose of this work was to compare adherence rates of M. bovis field strains to different host cell lines and study the effects of cloning and sub-culturing M. bovis strains on their adherence properties. Eighteen metabolically labeled M. bovis strains isolated from different pathological backgrounds were examined in adherence trials using four different host cell lines, i.e. embryonic bovine lung (EBL), embryonic bovine trachea (EBTr), Madin Darby bovine kidney (MDBK) and rabbit kidney (RK) cells. Although large interstrain variations in adherence rates (3.4-19.1%) were measured they could not be correlated to the pathological background (pneumonia, arthritis or mastitis). Adherence rates to the fibroblast cell line (EBTr) were significantly lower than those to the three epithelial cell lines (EBL, MDBK and RK). The only non-pathogenic strain (221/89) exhibited lower adherence rates than three isolates from clinical mastitis. Interestingly, adherence rates were significantly reduced after in vitro passaging. In contrast, no effect of single cloning of strains on adherence was observed. There was no general correlation between expression of variable surface proteins (Vsps) as monitored by immunoblotting and adherence rates, although alterations in Vsp expression profiles were seen as a consequence of passaging. As there is probably a large number of adhesins, variable and non-variable, on the surface of M. bovis cells the issue is very complex, and the most active components have yet to be identified.

The susceptibilities of 40 recent Belgian field isolates of Mycoplasma bovis to 10 antimicrobial agents were assessed. Tiamulin was the most active antimicrobial agent against M bovis, with an initial inhibitory concentration (IIC50) of 0.06 microg/ml, but it is not licensed for the treatment of cattle. All three fluoroquinolones tested (danofloxacin, enrofloxacin and marbofloxacin) were effective against strains of M bovis, and had a minimum mycoplasmacidal concentration (MMC50) less than or equal to 1 microg/ml. Gentamicin was poorly effective, having an IIC50 of 8 microg/ml. Many strains of M bovis were resistant to tylosin, spectinomycin, lincomycin, tetracycline and oxytetracycline.

Mycoplasmas frequently infect cattle. Amongst them, M. Bovis is the most pathogenic species in countries free from contagious bovine pleuropneumonia because it is responsible for bronchopneumonia, arthritis and mastitis, and is thus associated with strong economic losses. Several studies have shown the frequency of M. bovis in Europe and the spread of antibiotic-resistant strains. Considering the absence of vaccine in Europe, it is essential to understand this bacteria in order to control the infection in cattle. In this context, this paper aims at summarizing the current knowledge about M. bovis.

Mycoplasma bovis est frequemment isole en Belgique chez les bovins presentant des troubles respiratoires. Il est souvent associe aux pasteurelles et au virus respiratoire syncytial bovin, demontrant le caractere multifactoriel des pathologies respiratoires, la complexite de leur diagnostic, de leur prophylaxie et de leur therapie. En raison de la frequence elevee des souches multiresistantes, il est donc primordial d'optimaliser la prophylaxie soit par un vaccin soit par le respect des regles d'hygiene et de gestion des troupeaux. La comprehension des mecanismes d'adhesion de M. bovis aux cellules de l'hote est essentielle. Nous avons ainsi montre in vitro que l'adhesion est fonction (i) de la souche bacterienne, (ii) de son nombre de passages en culture et (iii) de la lignee cellulaire utilisee lors des tests d'adhesion. La souche de reference PG45 s'est averee non representative de la plupart des souches de terrain, probablement en raison du nombre eleve de passages subis en milieu de culture inerte. Enfin, M. bovis est capable d'adherer specifiquement sur des cellules epitheliales bronchiques bovines en culture primaire au moyen de plusieurs proteines dont les proteines variables de surface C et F.

Les mycoplasmes infectent frequemment les cheptels bovins. Parmi eux, Mycoplasma bovis est l'espece la plus pathogene dans les pays indemnes de peripneumonie contagieuse du bovin puisqu'elle provoque des bronchopneumonies, des arthrites et des mammites, et engendre des pertes economiques considerables. Plusieurs etudes ont montre sa frequence en Europe et l'acquisition d'antibioresistances. Vu l'absence de vaccin a l'heure actuelle en Europe, il est crucial de connaitre et de comprendre cette bacterie afin de controler l'infection chez les bovins. Dans ce but, cet article resume les connaissances actuelles sur M. bovis.

Mycoplasmas frequently infect cattle, causing especially respiratory diseases. Mycoplasma bovis is the most important pathogenic species in countries free of bovine contagious pleuropneumonia. This species was frequently isolated in Belgium from cattle with respiratory disease. Furthermore, associations were often observed with pasteurellas and bovine respiratory syncytial virus. Of these M. bovis isolates, many were resistant to several antimicrobial agents which are used in cattle practice, except to fluoroquinolones. Inasmuch the high frequency of M. bovis isolation and antibiotic resistances, it is very important to understand the pathogenicity of this bacteria in order to optimize prophylactic tools. Therefore, the study of the cytadherence of M. bovis is essential since it represents the first step of the bacterial infection. According to our experimental results, PG45 is not representative of field isolates because of its low adherence rates to various cell lines. This could be explained by the high number of subcultures of this pathogenic strain underwent since its first isolation, which sharply contrasts with other isolates. M. bovis adheres specifically to bovine bronchial epithelial cells in primary culture. Proteins such as variable surface proteins C and F are involved in this step as observed by decreased adherence rates after trypsinization of mycoplasma cells or addition of monoclonal antibodies.

Mycoplasma bovis is responsible for considerable economic losses in cattle due to pneumonia, arthritis and mastitis. As the agent was shown to be capable of adhering to neutrophils and embryonic bovine lung (EBL) cells and invading the respiratory epithelium it is highly desirable to improve our understanding of cytadherence processes. Although several surface proteins likely to be directly involved in this initial stage of interaction between pathogen and host cells have been identified, these findings mainly referred to type strain PG45 adhering to the continuous EBL cell line. The present study provides new and complementary data about cytadherence of M. bovis based on adherence of various radiolabeled strains to a primary culture of bovine bronchial epithelial (BBE) cells using a standardized adherence assay. M. bovis was shown to adhere specifically to the primary culture of BBE cells. Inhibition of adherence was observed upon addition of monoclonal antibodies (MAbs), trypsin treatment of mycoplasmas, and competition with non-radiolabeled mycoplasma cells. Interestingly, three MAbs against proteins involved in adherence to EBL cells failed to inhibit significantly the adherence to BBE cells. On the other hand, significant reduction of adherence rates by MAbs 2A8 and 9F1 directed against epitopes of variable surface lipoproteins VspC and VspF, respectively, demonstrated the involvement of these proteins in adherence of M. bovis to primary culture of BBE cells.

Between 1997 and 2000, a total of 150 healthy cattle and 238 animals with respiratory disease were examined for six Mycoplasma species. Attempts were made to detect Mycoplasma canis, Mycoplasma dispar and Ureaplasma diversum in calves with recurrent disease, and all three of these species were identified in calves with recurrent disease and in healthy lungs. In healthy calves, 84 per cent of bronchoalveolar lavage fluids were mycoplasma free; when cultures were positive, Mycoplasma bovirhinis was the only species isolated. Mycoplasmas were isolated from 78 per cent of animals suffering recurrent respiratory disease and from 65 per cent of acute respiratory cases. Mycoplasma bovis was isolated from bronchoalveolar lavages from 35 per cent of calves suffering recurrent respiratory disease, and from 50 per cent of acute cases, and from 20 per cent of pneumonic cases examined postmortem. M bovis was associated with other Mycoplasma species in 44 per cent of cases. M dispar was also isolated from 45.5 per cent of calves suffering recurrent respiratory disease, often in association with M bovis. M canis was identified for the first time in diseased Belgian cattle. Other mycoplasmas, including Mycoplasma arginini, Mycoplasma alkalescens and U diversum, were isolated less frequently. Associations between mycoplasmas and other pathogens were often observed. Among lungs infected with Pasteurella and/or Mannheimia species, more than 50 per cent were mixed infections with M bovis.

Three sampling procedures were compared to determine the optimal technique for isolating mycoplasmas in cattle with respiratory diseases. The prevalence of mycoplasmas isolated from these animals is also reported. In the first group, bronchoalveolar lavage (BAL) and nasal swab cultures were compared with the corresponding lung cultures from cattle necropsied for fatal respiratory diseases (n = 20). In a second group, nasal swabs were compared with corresponding BAL cultures in living animals with recurrent respiratory pathologies (n = 49). There was complete agreement between the paired BAL and lung cultures. In contrast, nasal cultures were not representative of the mycoplasmas present in the lower respiratory airways. The relative sensitivity and specificity of the nasal swab technique compared to BAL in living animals confirmed that the nasal swab cultures were not predictive of lower respiratory airway pathogens, such as Mycoplasma bovis. BAL is considered to be the best method for isolating M. bovis in cattle with respiratory diseases as it combines reliability and feasibility under field sampling conditions. In the present study, Mycoplasma dispar (43%) and M. bovis (29%) were mainly isolated in mixed infections. This confirms the need to search for mycoplasmas in routine examinations and to take them into account in therapeutic strategies for respiratory diseases in cattle.

The lower respiratory tract secretions can be useful in identifying microbial pathogens in case of pneumonia in calves. The transtracheal aspiration is the most frequently used technique to obtain these specimens but it is considered as traumatic (possible subcutaneous injuries). To avoid these complications, a simple and safe broncho-alveolar ravage technique, with obligation passage of the tube through the upper airways, was used and the results of bacteriologic culture of lower airway fluids obtained by use of both techniques were compared. The results obtained show that with a contaminated broncho-alveolar lavage fluid, the corresponding transtracheal aspiration fluid was either negative or bacteriologically less charged in 71% of the cases. These results clearly show that broncho-alveolar lavage are more contaminated that corresponding transtracheal aspiration. Thus, the broncho-alveolar ravage method by nasotracheal way does not reflect the real bacteriological pattern of the lower respiratory tract.

The purpose of this study is to reveal helicobacteria in a direct or indirect way. Biopsies came from the stomach of healthy or diseased dogs and cats submitted to a gastroscopical examination. Different steps and approaches have been followed: direct Gram staining, urease tests, culture on several appropriate media and comparison with histopathology. The results have shown that the stomach of carnivora in Belgium are colonized by helicobacteria. We have not been able to identify the species of the genus Helicobacter and to associate in a statistical way the presence of helicobacteria with lesions. The use of PCR will improve these results.