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Abstract

Objective—To evaluate disease in kittens inoculated with Bartonella henselae strain LSU16.

Animals—Eighteen 12-week-old specific-pathogenfree kittens.

Procedure—Kittens were inoculated with B henselae strain LSU16 or saline (0.9% NaCl) solution. Blood samples were collected from kittens on alternate weeks, and bacteremia, clinical signs, and antibody concentrations were monitored for 6 months after inoculation.

Results—Kittens developed raised, erythematous areas at the site of inoculation within 72 hours. Swelling peaked at 14 days and resolved by 28 days after inoculation. Fever had a biphasic pattern, with an episode of 1- to 3-days' duration beginning 6 to 7 days after inoculation followed by an episode of 3- to 8- days' duration beginning 11 to 13 days after inoculation. Kittens were bacteremic by day 14 with peak bacteremia at days 14 to 28. Strong antibody responses to B henselae were detected. Clinical disease resolved before bacteremia became undetectable, but signs of disease correlated with the highest degree of bacteremia. Regional lymphadenopathy also was evident.

Conclusion and Clinical Relevance—Clinical disease in kittens was similar to that in adult cats infected with B henselae strain LSU16, except that lethargy and anorexia were less severe in kittens, and a biphasic pattern of fever was detected in kittens. Clinical disease after inoculation with B henselae may be strain-dependent. To limit transmission of Bartonella organisms, appropriate flea prevention should be instituted.

Impact for Human Medicine—Kittens that are febrile, anorectic, lethargic, and that have lymphadenopathy should be tested for Bartonella organisms, and contact with immunocompromised owners should be discouraged. (Am J Vet Res 2000;61:375–379)

Full access
in American Journal of Veterinary Research

Abstract

Objective

To investigate the susceptibility of polarized epithelioid human rectal tumor (HRT-18G) cells to bovine coronaviruses (BCV) isolated from enteric (EBCV) and respiratory (RBCV) tract infections.

Procedure

Cells of the G clone of HRT-18 were grown to confluent monolayers on permeable supports, and were directionally infected at the apical and basolateral domains with 3 wild-type BCV strains, RBCV-LSU-94LSS-051-2, RBCV-OK-0514-3, and EBCV-LY138-2, and 1 cell culture-adapted strain, EBCV-L9-80. Sequential cytopathic changes were microscopically monitored. Medium samples for titration of hemagglutinins and viral infectivity were collected directionally from both domains of the infected cell cultures at various intervals.

Results

Polarized epithelioid HRT-18G cells from apical domains had maximal susceptibility to infection with the EBCV and RBCV strains, and those from basolateral surfaces had minimal susceptibility. Titers of hemagglutinins and infective progeny BCV reached 1,280 hemagglutinin units and 4.2 × 108 plaque-forming units/ml for apical samples, but were minimal for basolateral samples. Asymmetric virus release occurred through the apical surfaces of the HRT-18G cells by 12 hours after infection when cell fusion as a sign of cytopathic changes began. When cells were infected basolaterally, progeny virions released from apical surfaces reinfected the target cells from the apical domains and induced cytopathic changes were delayed about 12 hours, compared with changes detectable in apically exposed cultures.

Conclusions

EBCV and RBCV, isolated from cattle, had marked tropism for polarized epithelioid HRT-18G cells. Entry of BCV into the polarized HRT-18G cells was effected maximally through the apical domains and minimally through the basolateral domains. Release of progeny BCV occurred preferentially from the apical domains. (Am J Vet Res 1997;58:1120–1124)

Free access
in American Journal of Veterinary Research

Abstract

Objective

To develop a monoclonal antibody-based capture ELISA for detection of a 26- to 28-kd coproantigen of Fasciola hepatica in the feces of infected cattle.

Animals

27 crossbred yearling calves, 2 New Zealand White rabbits.

Procedure

A capture ELISA that uses a previously described monoclonal antibody (MAB) M2D5/D5F10 was developed. The MAB was used to capture the antigen from the feces, and hyperimmune rabbit serum raised against the purified 26- to 28-kd glycoprotein was used to detect the coproantigen. This test was used for the detection of the antigen in the feces of 27 experimentally infected calves with known numbers of flukes. Fecal specimens obtained before infection from the same calves were used as negative controls.

Results

The assay results identified all calves infected with more than 10 flukes at necropsy, and as little as 300 pg of coproantigen/ml of fecal supernatant was detected. The assay results correlated well with the number of flukes, suggesting that it is possible to estimate fluke burden. Infections as early as 6 weeks’ duration were detected, before flukes mature to adults and start to shed eggs.

Conclusions

In experimentally infected calves, the coproantigen capture ELISA was more sensitive and easier to perform than microscopic examination for the diagnosis of F hepatica infection; moreover, 6-week-old prepatent infections were detectable.

Clinical Relevance

This capture ELISA containing an F hepatica 26- to 28-kd coproantigen is a quantitative assay that is more sensitive than fecal egg counting. In addition, the assay is rapid, easy to perform and lends itself well to large numbers of samples. Because it is antigen based, the ELISA may be useful for diagnosis of F hepatica infection in other species, including human beings. (Am J Vet Res 1998;59:533–537)

Free access
in American Journal of Veterinary Research

Abstract

Objective

To investigate the antigenic diversity of lipo-oligosaccharides of Haemophilus parasuis.

Procedures

Immunoblot assays were done with monoclonal and polyclonal antibodies on whole-cell lysates. Individual colonies of H parasuis strains H 54, H 53, and H 128 were tested for reactivity with lipo-oligosaccharide-specific monoclonal antibodies after a single passage on chocolate agar, and colonies of strain H 54 were analyzed after 10 passages. Colony blot tests were used to screen H parasuis strains for spontaneously occurring antigenic variation in their lipo-oligosaccharides.

Results

Eight H parasuis strains were separated into 4 lipo-oligosaccharide serovars on the basis of immunoblot reactions with 3 polyclonal rabbit antisera. Nine monoclonal antibodies against lipo-oligosaccharides of a lipo-oligosaccharide-serovar I strain reacted with all tested serovar I strains but failed to react with other H parasuis strains.

Conclusions

Variations in the antigenic reactivity after 1 or 10 passages on chocolate agar were not observed. The serovar I lipo-oligosaccharide strains included virulent as well as avirulent H parasuis strains, indicating that these epitopes do not correlate directly with virulence properties of H parasuis. (Am J Vet Res 1996;57:63-67)

Free access
in American Journal of Veterinary Research