Search

The lipolytic enzyme hepatic lipase (HL) may facilitate mobilization of cholesterol substrate for ovarian steroidogenesis. We investigated whether HL was necessary for optimum reproduction in the female mouse by analyzing breeding performance and ovarian responses to gonadotropins in HL-/- mice. HL-/- female mice bred with HL-/- males had the same pregnancy success rate and pup survival rate as did wild-type (WT) mice but had significantly smaller litters, producing 1.7 fewer pups per litter. Mice were primed with eCG/hCG, and at 6 h post-hCG the HL-/- mice had smaller ovaries than did the WT mice. HL deficiency specifically affected ovarian weight; adrenal gland weights did not differ between WT and HL-/- mice. HL-/- mice weighed more than age-matched WT mice. Between the two mouse genotypes, uterine weights were the same, indicating that estrogen production was equivalent. However, the HL-/- ovaries produced significantly less progesterone than did the WT ovaries within 6 h of hCG stimulation. HL-/- ovaries had the same number of large antral follicles as did the WT ovaries but had fewer hemorrhagic sites, which represent ovulations, fewer corpora lutea, and more oocytes trapped in corpora lutea. We suggest that reduced progesterone synthesis following hCG stimulation attenuated the final maturation of preovulatory follicles, resulting in smaller ovaries. Furthermore, reduced progesterone production limited the expression of proteolytic enzymes needed for tissue remodeling, resulting in fewer ovulations with a corresponding increase in trapped or unovulated oocytes and providing a possible explanation for the smaller litter size observed in spontaneously ovulating HL-/- mice.

Clostridium piliforme is an obligate intracellular bacterium that causes enterohepatic disease in many animal species. C. piliforme infections are commonly subclinical in laboratory rats and mice, and little is known about host regulation of disease or of the effects of C. piliforme infections on investigations that use subclinically infected animals. To assess host regulation of subclinical C. piliforme infections and the effects of those infections on laboratory mice, the expression of the pro-inflammatory cytokines tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) was evaluated at 0, 1, 3, 7, 14 and 28 days after inoculation with C. piliforme. Subclinical infection was induced in weanling C. piliforme-susceptible DBA/2 or -resistant C57BL/6 mice with either a toxic or a non-toxic C. piliforme strain. Hepatic lesions and bacteria were demonstrated histologically in both mouse strains for 14 days after inoculation with the toxigenic bacterial strain, but were never demonstrated histologically following inoculation with the non-toxigenic strain. Hepatic TNF-alpha and IFN-gamma mRNA and serum protein levels were similarly elevated in both mouse strains 1 day after inoculation with both C. piliforme strains, as evaluated by reverse transcription PCR and enzyme-linked immunosorbent assays, respectively. Elevation of IFN-gamma persisted for 14 days after inoculation; TNF-alpha remained elevated at 28 days after inoculation.

Clostridium piliforme induces enterohepatic disease in many domestic and laboratory animal species. Susceptibility to infection is known to vary with the immune status and strain of the host, but little is known about specific immune mechanisms that regulate this disease. To evaluate host control of C. piliforme infection, we examined the role of interleukin-12 (IL-12) both in the control of and in the response to murine C. piliforme infection. For this study, 3-week-old C. piliforme-resistant C57BL/6 or -susceptible DBA/2 mice were infected intravenously with either the toxic H1 or the nontoxic M1 C. piliforme isolate. Serum and liver samples were collected prior to C. piliforme inoculation (day 0) and at days 1, 3, 7, 14, and 28 postinoculation. Evaluation of hepatic IL-12 p40 mRNA expression by reverse transcription-PCR and of total-IL-12 protein levels in serum by enzyme-linked immunosorbent assay revealed that C. piliforme induced elevations in both hepatic p40 mRNA and serum total-IL-12 levels at all times postinoculation. Elevations were similar with both toxic and nontoxic C. piliforme isolates. Levels of total IL-12 in serum were significantly (P < 0.05) higher in C57BL/6 mice than in DBA/2 mice. Additional experiments were performed in which polyclonal antibody treatment was used to neutralize IL-12 in mice of both strains prior to intravenous inoculation with toxic C. piliforme H1. IL-12 neutralization increased the severity of Tyzzer's disease at day 3 postinoculation in both mouse strains, but the degree of increase was greater in C57BL/6 mice than in DBA/2 mice.

Clostridium piliforme infection (Tyzzer's disease) induces enterohepatic disease in many domestic and laboratory animals. Murine susceptibility to Tyzzer's disease varies with host strain, age, and immune status However, little is known about the role of the immune system in control of this disease. To investigate the role of host immunity in Tyzzer's disease, mice were depleted of either neutrophils, natural killer cells, or macrophages by antibody administration or chemotherapy. After depletion, DBA/2 mice, which are naturally susceptible to C. piliforme, or naturally resistant C57BL/6 mice were inoculated intravenously with C. piliforme. Animals were euthanized 3 days postinoculation and evaluated for gross and histologic lesions and hepatic bacterial load. In juvenile DBA/2 or C57BL/6 mice, depletion of either neutrophils or natural killer cells increased severity of disease. In adult mice, depletion of natural killer cells significantly increased severity of Tyzzer's disease in the resistant (C57BL/6) but not in the susceptible (DBA/2) strain. Macrophage depletion did not alter the course of infection in either mouse strain. These studies indicate an important role for neutrophils and natural killer cells in the pathogenesis of murine Tyzzer's disease. The role of macrophages in murine C. piliforme infection will require further evaluation.

Ninety-seven BALB/c mice were inoculated intravaginally with 8.0 x 10(5) Tritrichomonas foetus organisms, using either isolate ATCC 30003 or field isolate MU Y22 2 days after estrogenization with 15 micrograms 17 beta-estradiol. Reproductive tracts were examined at several time points post-inoculation to determine gross and histologic responses to trichomonad infection as compared to estrogenized, uninfected control animals. The two isolates varied greatly in ability to maintain chronic infection; no ATCC 30003-inoculated animals remained culture-positive beyond 7 weeks post-inoculation, whereas MU Y22-inoculated animals were infected for greater than 26 weeks. Lesions were seen in 40-60% of animals prior to 10 weeks post-inoculation and included moderate uterine dilation and glandular atrophy, uterine gland abscesses, pyometra, intramural perivascular lymphoid infiltrates, and ovarian bursitis. The severity of lesions was independent of the T. foetus isolate. Lesions became more severe at 10 weeks post-inoculation, and at 10 and 26 weeks post-inoculation, lesions were seen in 60% and 75% of animals, respectively. In addition to lesions described above, epithelial changes were marked at these late necropsies, including ulceration, flattening, hypertrophy, and squamous metaplasia. The lesions seen in these mice closely resemble those described in natural bovine infection, suggesting that the estrogenized BALB/c mouse is an excellent model for study of bovine trichomoniasis.

Four guinea pigs from a colony of approximately 50 animals were examined for progressive neurologic disease of 5 days' duration. Signs of neurologic dysfunction included cachexia, stupor, hyperexcitability, lateral recumbency, and opisthotonos. Results of gross pathologic, microbiologic, and serologic examinations were unremarkable. Histologic examination of cerebral and cerebellar sections revealed multifocal malacia and regions of eosinophilic granulomatous inflammation. Cross-sections of nematode larvae, identified as Baylisascaris sp., most likely B. procyonis, the raccoon ascarid, were seen in the brain of some affected animals. An intact Baylisascaris larva was recovered from a symptomatic animal when cerebral tissue was processed by the Baermann extraction technique. Results of further investigation indicated that wood shavings used for the guinea pigs had been contaminated by raccoon feces, some of which contained numerous B. procyonis eggs. The bedding source for this colony was changed and, to date, no new cases of neurologic disease have been seen. This report emphasizes the potential insidious entrance of B. procyonis into well-managed laboratory animal facilities.