A single dose of vaccine for Mycoplasma bovis pneumonia, inactivated with saponin, was inoculated subcutaneously into 3-4 week-old calves. The calves were challenged 3 weeks later with a virulent strain of M. bovis on two occasions within 24h using the aerosol route. The calves were monitored for clinical signs and serological responses then post mortemed 3 weeks after challenge. The vaccine was shown to be highly immunogenic in calves and did not cause adverse effects. Vaccinated calves showed few clinical signs while all unvaccinated calves developed signs of pneumonia. There was a significant decrease in body weight gain in unvaccinated calves compared to vaccinates and a significant increase in lung lesions and rectal temperatures in unvaccinated calves. The vaccine also reduced the spread of M. bovis to internal organs. In conclusion the M. bovis vaccine produced a significant level of protection against a large virulent challenge.

SUMMARYThe spread of inflammation of the cloaca and phallus in goose flocks was investigated. In the flocks surveyed, 57.5 to 71.8% of the initial gander stock and 39.8 to 100% of the replacement ganders were affected. Based on the results of several hundred attempts at mycoplasma isolation, a relationship was found between mycoplasma infection and the occurrence of inflammation of the cloaca and phallus in the flocks. Mycoplasmas were frequently isolated from the mucous membrane of the phallus, the pallic lymph, and the inner organs of ganders in the affected flocks, but not from birds in unaffected flocks. Two biochemically (glucose-splitting and arginine-hydrolysing) and three serologically distinct types of mycoplasmas have been isolated. One of them proved to be identical to strain 46, identified by J.M. Bradbury as M. cloacale, and two strains may represent new Mycoplasma species since they differ serologically from all previously known Mycoplasma spp. In the diseased flocks, the strain designated 1220 (8390) and the antibodies produced against it were of most frequent occurrence. At the peak of egg production, mycoplasmas were isolated from 92.1% of the phallic lymph samples, and 94.6% of the test sera were positive for antibodies to strain 1220 (8390).

The GenBank accession numbers for the 16S rRNA gene sequence and the genome of the strain 1220 T are EU597750 and CP042295, respectively, and the GenBank accession numbers for other housekeeping genes used for the genetic characterization of each novel species described in this study are provided in the manuscript. †These authors contributed equally to this work Two supplementary figures, two supplementary tables and one supplementary document are available with the online version of this article.

A Mycoplasma bovis species-specific PCR assay has been developed with improvement of a previously described method (Ghadersohi et al., 1997). This test and its semi-nested version (Hayman and Hirst, 2003) did not function at all in our hands. A new reverse primer (Mbr2) was designed using previously published sequence data. For testing specificity, DNA was extracted from the most frequently occurring mycoplasma species and bacteria of bovine origin. The new PCR detected only Mycoplasma bovis. Moreover, no cross-reaction was observed with the genetically closest relative species, M. agalactiae. The target organism could be detected in a dose as low as 150 CFU ml -1 in broth cultures using ethidium-bromide-stained agarose gels.

SUMMARYInfertile eggs, dead embryos and tissues from laying geese (airsacs, peritoneum, oviduct, ovary, ova) were examined for presence of mycoplasmas. Forty-three of 110 eggs and the birds laying mycoplasmacontaining eggs proved to be positive for mycoplasmas. One of the strains was used for experimental infection of laying geese. A reduction in egg production, an increased number of infertile eggs, egg transmission of mycoplasmas and loss of body weight of hatched goslings, were observed due to the mycoplasma infection.

Bacterial compounds signal the presence of foreign pathogens in the innate immune system. These microbial components are key players in infectious diseases and implicate toll-like receptors in the activation of inflammation and coagulation. Nevertheless, the existence of a synergistic relationship between peptidoglycan and bacterial DNA on these two physiological responses has not been investigated. The present study reports new findings on the regulation of tumor necrosis factor alpha and tissue factor in peripheral blood mononuclear cells by peptidoglycan and bacterial DNA. These were found to induce tumor necrosis factor alpha and tissue factor simultaneously and in a synergistic manner. These findings provide a new proinflammatory and procoagulant mechanism likely to play a role in sepsis pathogenesis.

We performed a comparative study on the development of mastitis induced by Mycoplasma arginini or Streptococcus dysgalactiae after challenging the cows. Mycoplasma arginini did not cause any clinical symptoms on its own, resulting in only a transient increase of somatic cell count (SCC; increase ranging from 0.5 × 10(6) to 0.8 × 10(6) cells/mL) and a slight decrease of milk production (10%) for 5 d. In contrast, Strep. dysgalactiae induced more severe clinical signs in animals and SCC increased to 1.60 × 10(6) to 2.11 × 10(6) cells/mL for 10 d. In addition, milk production decreased (22.9 to 27.0%) for 10 d. After 3 mo (2 mo after the first challenge), animals that were challenged previously with M. arginini were rechallenged with Strep. dysgalactiae. Severe clinical mastitis developed, with very high SCC (5.00 × 10(6) to 21.5 × 10(6) cells/mL), and a very significant reduction of milk production (28.6 to 68.7%), which lasted more than 4 wk, was observed. The severe clinical mastitis developed not only in cows inoculated with Strep. dysgalactiae andM. arginini in the same udder quarter but also in cows infected in the quarter previously not challenged with mycoplasma. Cows challenged first with Strep. dysgalactiae and rechallenged with M. arginini 2 mo later developed only slight changes in both SCC and milk production, similar to those when the cows were challenged with M. arginini alone. We conclude that M. arginini infection does not cause remarkable mastitis (characterized by decrease in milk production and increase of SCC) but it significantly predisposes animals to infection with Strep. dysgalactiae, leading to severe clinical mastitis.

Phenotypic and genotypic diversity of forty-two Pasteurella multocida isolates from geese were characterized by analysis of their capsular type, Heddleston serotype, biotype, fimbrial gene allele type, comparative outer membrane protein (OMP) electrophoresis patterns, and were analyzed using PCR for the presence of virulence-associated genes (toxA, tbpA, pfhA, hgbA, hgbB, nanH, nanB, fimA, hsf-1, and pmHAS). A sequence comparison of the thdF and rpoB housekeeping genes of twenty representative P. multocida strains from three different OMP groups demonstrated that seventeen strains were closely related phylogenetically to previously published strains of P. multocida subsp. multocida and P. multocida subsp. gallicida, and only three strains from geese were grouped with previously published strains of P. multocida subsp. septica. This study is the first report regarding the characterization of phenotypic and genotypic features of different P. multocida field strains isolated from geese with the different clinical representation of pasteurellosis and provide our overview on the usefulness of these in vitro tests for primary characterization of P. multocida strains for the epidemiological studies of fowl cholera.