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Biomedical subjects

S Tagliabue

Publications and source records attributed to S Tagliabue.

9 recordsLinked to original sources

Diagnosis of Mycobacterium bovis infection in calves sensitized by mycobacteria of the avium/intracellulare group.

Because of the frequent exposure of cattle to mycobacteria of the avium/intracellulare group, an investigation was carried out into the possible repercussions thereof on the diagnosis of bovine tuberculosis. Three calves from a bovine tuberculosis-free herd, scored avian reactors in the gamma-interferon assay for bovine tuberculosis, were sedated and inoculated endotracheally with a virulent Mycobacterium bovis strain. Then, three other avian reactors were housed with the above donor calves. Mycobacterium bovis was isolated from the nasal swabs of the three endotracheally infected, donor calves. On these samples, TB complex-specific polymerase chain reaction (PCR) tests for IS6110 were also positive, albeit with a different time kinetics. The three contact-infected calves showed clear immunological signs of infection; however, their nasal swabs were always PCR-negative and only Mycobacterium avium was isolated. In the endotracheally infected donor calves there was a rise of the gamma-interferon responses to avian and bovine purified protein derivative (PPD) tuberculins, which reached the same stable plateau levels over the whole experiment. The above effect was also observed in the contact-infected calves, even though the response to avian PPD tuberculin always remained at a higher level. By using conventional bovine and avian PPD tuberculins, the comparative intradermal test was generally positive in endotracheally infected, as opposed to contact-infected calves; a positive intradermal test for M. bovis was obtained in two contact-infected calves by different bovine PPD tuberculins based on M. bovis bacillus Calmette-Guerin (BCG) secreted or somatic antigens. It was concluded that M. bovis infection may be concealed for some time in cattle sensitized by mycobacteria of the avium/intracellulare group and that different diagnostic procedures should be adopted for such animals.

Animals↗

Serological survey of leptospiral infection in kennelled dogs in Italy.

Two hundred and forty-five dogs were examined serologically for the presence of antibodies against different serovars of Leptospira interrogans. The dogs belonged to five different groups: group 1 was composed of clinically healthy pet dogs referred for a regular veterinary check-up visit or for vaccination; group 2 was composed of stray dogs; and groups 3, 4 and 5 were composed of dogs maintained in three different kennels which had varying standards of hygiene. Seventy-two out of the 245 dogs examined were seropositive for leptospirosis. In group 1, there were 3-4 per cent seropositive dogs; in group 2, 30.3 per cent; in group 3, 13.8 per cent; in group 4, 38.6 per cent; and in group 5, 49.2 per cent. This study demonstrates that leptospiral infection is common in dogs housed in kennels, despite most of them being vaccinated, and that crowding of animals into unsanitary quarters is associated with a high prevalence of infection. The most common infecting serovars found were bratislava and grippotyphosa, confirming recent observations that demonstrate a significant change in the epidemiology of canine leptospirosis.

Animals↗

Quality controls and in vitro diagnostic efficiency of bovine PPD tuberculins.

Wide heterogeneity was shown among different batches of bovine protein purified derivative (PPD) tuberculin, as regards protein content and antigenic profile. These features were also investigated in pilot preparations of Mycobacterium bovis secreted antigens and PPD tuberculins. Under controlled conditions, widely different compositions were revealed as a function of the M. bovis strain and also of the time in culture, due to the transient expression of seemingly important clusters of antigens. Furthermore, these parameters could dramatically affect the efficacy of the above preparations in in vitro assays of cell-mediated immunity on M. bovis-infected cattle. The field exposure to mycobacteria of the avium/intracellular group could also influence the readout of such assays, results being in agreement with a bystander suppression model of the response to M. bovis antigens. Due to the above, practical suggestions are put forward to improve the composition of bovine PPD tuberculins and the relevant control procedures.

Animals↗

Leptospira interrogans serovar sejroe infection in a group of laboratory dogs.

Interstitial nephritis was seen histologically in 19 (59%) out of 32 pure-breed beagle dogs (16 males and 16 females) subjected to standard safety tests. In these animals no clinical abnormalities were observed and all the tested parameters (haematology, biochemistry and urine analysis) were within the normal ranges. Leptospiral antibody titres ranging from 1 : 100 to 1 : 6400, against a serovar (hardjo) belonging to the Sejroe serogroup, were detected by the microscopic agglutination test (MAT) in the serum of the 19 dogs with interstitial nephritis. All animals without renal lesions were seronegative. Leptospiral antigen was detected immunohistochemically in the kidneys of 4 dogs; leptospires were detected in Warthin-Starry stained sections of one dog. Leptospires were isolated from the kidneys of 3 of the 4 dogs examined by bacterial culture. The isolated strains were typed as serovar sejroe by restriction endonuclease digestion and Southern blot hybridization analysis of their DNA. It was concluded that Leptospira interrogans serovar sejroe, was responsible for an asymptomatic chronic renal infection which was widespread in this group of laboratory dogs.

Agglutination Tests↗

Detection and identification of Leptospira interrogans serovars by PCR coupled with restriction endonuclease analysis of amplified DNA.

Primers for PCR were selected from a sequenced fragment of clone pL590, which contains a repetitive element present in the genome of Leptospira interrogans serovar hardjo type hardjoprajitno (M. L. Pacciarini, M. L. Savio, S. Tagliabue, and C. Rossi, J. Clin. Microbiol. 30:1243-1249, 1992). A specific DNA fragment was amplified from the genomic DNAs of serovar hardjo type hardjoprajitno and nine serovars also belonging to L. interrogans as a consequence of the spread of the same or a closely related repetitive element within this species (Pacciarini et al., J. Clin. Microbiol. 30:1243-1249, 1992). In addition, specific amplification was obtained from two Leptospira borgpetersenii serovars (tarassovi and hardjo type hardjobovis). Negative PCR results were observed with all of the other Leptospira serovars tested, including nonpathogenic ones (serovars patoc and andamana), another spirochete (Borrelia burgdorferi), bacteria commonly found in biological samples, and swine and bovine cell lines. Direct PCR on biological samples such as kidney samples demonstrated that preliminary isolation and culture of Leptospira cells are not required for efficient detection. Furthermore, digestion of the amplified DNA with the enzymes HinfI and DdeI yielded specific polymorphic patterns, allowing discrimination among the majority of the serovars. These methods were applied to 25 field isolates of serovar pomona, leading to the conclusion that they were suitable for the simple and rapid detection of L. interrogans and for serovar identification.

Animals↗

The search for improved methods for diagnosing leptospirosis: the approach of a laboratory in Brescia, Italy.

The authors describe work in progress at the laboratory in Brescia, Italy, on the application of molecular methods to the diagnosis of leptospirosis. This work includes the following: a) Development of polymerase chain reaction (PCR) assays capable of amplifying specific deoxyribonucleic acid fragments from most Leptospira interrogans strains. b) Development of a microtitre-based assay for rapid detection of PCR-positive samples. c) Characterisation of Leptospira strains through restriction endonuclease analysis of PCR products and amplified fragment length polymorphism.

Animals↗

Identification of Leptospira interrogans strains by monoclonal antibodies and genomic analysis.

A recombinant probe derived from a genomic library of serovar hardjo strain Hardjoprajitno, and a panel of serovar specific Monoclonal Antibodies (MAbs) were used for the characterization of 31 Leptospira isolates from cattle and swine. The two methods performed equally well in serovar identification except for the distinction of the genotypes hardjoprajitno and hardjobovis within serovar hardjo which could only be obtained by genomic analysis. The combination of immunological and genetic information was also useful to evaluate the degree of variability of Leptospira strains. The quality of the patterns and the sensitivity provided by a digoxigenin labelled probe were comparable to those obtained with a radioactive reagent.

Animals↗

Repetitive sequences cloned from Leptospira interrogans serovar hardjo genotype hardjoprajitno and their application to serovar identification.

We selected, from a genomic library of Leptospira interrogans serovar hardjo genotype hardjoprajitno, two probes containing repetitive sequences (pL1 and pL590). The hybridization patterns of these probes to DNA isolated from a variety of Leptospira serovars were examined and their ability to detect subtle differences at the genomic organization level was established. We identified the DNA fragments within pL1 and pL590 which are sufficient to yield polymorphic hybridization patterns; these results define the upper size limit of two novel repetitive elements in the Leptospira genome. The pattern and degree of hybridization observed for the serovars tested in this work were used to divide Leptospira spp. into groups which share genetic relatedness; our conclusions are consistent with previous classifications by other authors.

Blotting, Southern↗