Larval migration of the ascarid nematode Toxocara canis following infection and re-infection in the gerbil Meriones unguiculatus.

A morphological and immunohistochemical study of larval migration patterns was performed in gerbils that were infected once (primary infected group) or twice (secondary infected group) with 1500 eggs of Toxocara canis. Animals from the primary infected and the re-infected group were killed at different times after infection, and larvae were counted in the intestines, liver, lungs and brain. Fragments of all organs were formalin fixed and paraffin embedded for histology and immunohistochemistry analyses (using polyclonal anti-Toxocara serum raised in rabbits infected with T. canis). In the primary infected group, larvae were more abundant in the intestine at 24 h, in the liver and lungs between 24 and 72 h and in the brain after 96 h; larvae predominated in the brain for up to 60 days after infection. In the re-infected group, an increase in the number of larvae in the liver and a reduction in the number of larvae in the brain was observed up to 60 days after re-infection. Inflammatory reactions were absent or limited. Eosinophils and loose granulomata were observed around the larvae and their antigens in the primary infected group and were more severe. Many eosinophils and typical epithelioid granulomata were observed around larvae in the re-infected group. These results demonstrate that the migration pattern of T. canis larvae in gerbils is similar to that in mice and rats, exhibiting a late neurotropic stage. In the re-infected group, there was histological evidence of an adaptive T-helper 2 (Th-2) response, and larvae were apparently retained within granulomata in the liver, without obvious signs of destruction.

PCR-RFLP genotyping of Toxoplasma gondii from chickens from Espírito Santo state, Southeast region, Brazil: new genotypes and a new SAG3 marker allele.

Brazil is one of the regions with the highest prevalences of Toxoplasma gondii in humans and animals. Because free-range chickens become infected by feeding from ground contaminated with oocysts, the prevalence of T. gondii in this host has been widely used as an indicator of the strains prevalent in the environment. The genetic variability among T. gondii isolates from different healthy and sick hosts all over the world has been recently studied. Three clonal genetic lineages (Types I, II and III) were initially recognised as predominant in Western Europe and the United States. T. gondii strains are genetically diverse in South America. In Brazil, recombination plays an important role in strain diversification. The objective of this study was to genetically characterise T. gondii isolates from free-range chickens from Espírito Santo state, Southeast region, Brazil, using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). A total of 44 isolates among 47 previously described isolates (TgCkBr234-281) from free-range chickens were included in this study. Strain typing was performed using 12 PCR-RFLP markers: SAG1, SAG2, alt. SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico and CS3. Eleven genotypes were identified. Ten isolates (23%) were grouped into four novel genotypes. Four isolates, distributed in four counties, corresponded to the Type BrI lineage, the genotype found most frequently in Brazil. No clonal Types I, II or III lineages were found. Two novel genotypes were represented by single isolates. Unique alleles were identified for the markers SAG1, c22-8 and CS3, and for the first time, a unique allele was found for the marker SAG3. Although a large number of T. gondii genotypes have already been identified from a variety of animal hosts in Brazil, new genotypes are continuously identified from different animal species. This study confirmed the diversity of T. gondii in Brazil and demonstrates clonal Type I, II and III lineages are rare in this country.

Seroprevalence and isolation of Toxoplasma gondii from free-range chickens from Espírito Santo state, southeastern Brazil.

Prevalence of Toxoplasma gondii infection in 510 free-range (FR) chickens (380 from 33 small farms, and 130 from a slaughter house for FR chickens) from Espírito Santo state, southeastern Brazil, was investigated. Antibodies to T. gondii were sought using commercial indirect haemagglutination (IHAT, Imuno-HAI Toxo(®), Wama Diagnóstica, São Paulo, Brazil, cut-off 1:16) and the modified agglutination test (MAT, cut-off 1:25) tests. Attempts were made to isolate viable T. gondii from seropositive chickens by bioassay in mice. Pooled samples of brain, heart and quadriceps muscle of one thigh (total 40 g) from 64 chickens with IHAT titers of ≥ 1:16 were minced, digested in pepsin and bioassayed in mice. Antibodies to T. gondii were found in 40.4% (206/510) FR chickens by IHAT (titer ≥ 1:16) and 38.8% (198/510) by MAT (titer ≥ 1:25); concordance between IHAT and MAT was 81.6% (kappa index=0.614). Viable T. gondii was isolated (designated TgCkBr234-281) from 48 of 64 (75%) seropositive (IHAT titers ≥ 1:32) FR chickens. Most isolates of T. gondii were virulent for mice; 100% of mice inoculated with 44 of 48 isolates died of toxoplasmosis within 30 days post inoculation (p.i). An epidemiological investigation revealed that people living in rural areas have little knowledge about the parasite and about the risk of acquiring it from raw meat. Results indicated that the locally available IHAT was useful for screening of chicken sera for T. gondii antibodies.

Presence of intestinal helminths decreases T helper type 1 responses in tuberculoid leprosy patients and may increase the risk for multi-bacillary leprosy.

Resistance to intracellular pathogens such as Mycobacterium leprae is dependent upon an effective T helper type 1 (Th1)-type immune response. On the other hand, intestinal helminths are known to subvert the host's immune response towards to either a Th2-type immune response or a regulatory T cell up-regulation, which may affect the host's ability to mount an effective response to mycobacteria. Here, we report a significant association between intestinal helminth infections and lepromatous leprosy [odds ratio (OR), 10.88; confidence interval (CI) 95%: 4.02-29.4; P<0.001]. We also observed that the frequency of intestinal helminths correlated strongly with the mycobacterial index (r=0.982, P<0.01). Corroborating with our hypothesis, intracellular levels of interferon-gamma were decreased significantly in leprosy patients co-infected with intestinal helminths when compared to leprosy patients without worms. Conversely, lepromatous leprosy patients with intestinal worms produced higher levels of both interleukin (IL)-4 and IL-10. Our results suggest that a pre-existing infection by intestinal helminths may facilitate the establishment of M. leprae infection or its progression to more severe forms of leprosy.

The effect of buserelin acetate on the uterus of adult rats: morphological aspects.

PURPOSE OF INVESTIGATION: To evaluate the effect of buserelin acetate on the morphology of the endometrium of adult, non-castrated, female Wistar rats. METHODS: Female Wistar rats at estrus or diestrus (assessed by vaginal cytology) received daily subcutaneous injections of 20 mg buserelin acetate for four, eight or 12 days. Rats were sacrificed 24 hours or five days following final dosage. A control group received diluent for 12 days. RESULTS: Progressive tissue hypotrophy occurred during treatment and was followed by estrogenic hyperactivity five days after the end of treatment. Vaginal cytology and endometrial histology revealed intense, vacuolized lining and glandular epithelia, brush borders and endometrial stroma densely infiltrated with eosinophils. CONCLUSIONS: Buserelin acetate appears to cause a progressive blockade of gonadotrophin secretion when administered to female rats for four, eight or 12 days, and an important rebound effect, with accentuated estrogen release already apparent in the first estrous cycle following treatment.

Effects of simultaneous treatment with estrogen and testosterone on the uterus of female adult rats.

BACKGROUND: Testosterone (T) associated with estrogen (E) has been used in hormonal replacement therapy in postmenopause women and the effects of this hormonal association on the uterus are not known. OBJECTIVE: To study the effect of long-term simultaneous exposure to testosterone and estrogen on the uterus of non-castrated adult female rats. METHODS: Groups of ten adult noncastrated female Wistar rats were treated with non-esterified testosterone and beta estradiol (subcutaneous implants with 50 mg of each hormone) or with testosterone cipionate and estradiol valerate (weekly intramuscularly or by subcutaneous injection of respectively, 2.85 mg/kg and 0.166 mg/kg). Control groups received no treatment (10 rats) or injections of diluents (6 rats). All animals were killed six months after hormonal exposure. RESULTS: All rats treated with T+E developed hyperplasia and hyperkeratosis of the vaginal and cervical epithelium and focal metaplasia with keratinization of the endocervical and endometrial epithelium. Ascending pelvic inflammatory disease with pyometra and tuboovarian abscesses were frequent (25% mortality until the end of the experiment). CONCLUSIONS: Testosterone associated with estrogen induced metaplasia of the genital epithelium but did not induce neoplastic lesions. The metaplasic lesions reduced the mucosal defense mechanisms enhancing ascending genital inflammatory disease. Although metaplasia of the cervical and endometrial epithelium has been observed after estrogen exposure in rats, testosterone does not appear to inhibit these estrogen effects.

Toxocara infection is not associated with viral or bacterial central nervous system infection in children.

BACKGROUND: Helminth infections with larvae that migrate through the tissues have been considered risk factors for CNS infections. OBJECTIVES: The present work was designed to investigate the prevalence of anti- TOXOCARA antibodies in the serum and/or in the cerebrospinal fluid (CSF) of children with infectious meningitis or meningoencephalitis and of a control group, without meningitis, admitted at the Children's Hospital NS Glória, Vitória, ES, Brazil. PATIENTS AND METHODS: After adsorption with ASCARIS LUMBRICOIDES antigen, serum and/or cerebrospinal fluid of 381 inpatients (201 with meningitis and 180 without meningitis) were submitted to an ELISA IgG, for anti- TOXOCARA antibodies using secretion/excretion antigens of third stage larvae of T. CANIS. RESULTS: No significant differences between the meningitis and the control groups were observed in the frequencies of positive tests for anti- TOXOCARA antibodies in the serum or CSF (respectively for the meningitis and control group: 33/103 or 32 % and 52/152 or 34.2 % for the serum, p = 0.821; 48/184 or 26.1 % and 23/121 or 19.0 % for the CSF; p = 0.196. CONCLUSION: The results demonstrated that TOXOCARA infection, evaluated by detection of anti- TOXOCARA antibodies in serum or CSF, is not associated with viral or bacterial meningitis or meningoencephalitis in children in our country.

Morphological changes induced by testosterone in the mammary glands of female Wistar rats.

Increased levels of androgens in postmenopausal women are considered to be a risk factor for breast cancer. Testosterone, alone or in combination with estrogen, induces epithelial dysplasia and mammary tumors in Noble rats. Since this model of hormone-induced neoplasia has not been reported in other rat strains, we studied the effect of testosterone on the mammary gland morphology of female Wistar rats. Sixty adult, non-castrated, female Wistar rats were implanted in the dorsum midline with a silicone tube containing 50 mg testosterone (testosterone propionate in 30 animals and non-esterified testosterone in the remaining 30 animals) and 20 additional animals were implanted with empty tubes and used as control. Five animals per group were killed 30, 60, 90, 120, 150, and 180 days after implantation, and the mammary glands were dissected, fixed and embedded in paraffin. Histological sections were then stained with hematoxylin and eosin and picrosyrius red for collagen visualization. Morphological and morphometric analysis demonstrated ductal proliferation and acinotubular differentiation with secretory activity in all treated animals, peaking at 90 days of androgen exposure. After 90 days the proliferation of acinar epithelial cells was evident, but there was a progressive reduction of secretory differentiation and an increase in intralobular collagen fibers. There was no morphological evidence of dysplastic changes or other pre-neoplastic lesions. Testosterone treatment applied to adult, non-castrated female Wistar rats induced a mammary gland hyperplasia resembling the lactating differentiation, with progressive reduction in secretory differentiation.

Morphological changes induced by testosterone in the mammary glands of female Wistar rats

Increased levels of androgens in postmenopausal women are considered to be a risk factor for breast cancer. Testosterone, alone or in combination with estrogen, induces epithelial dysplasia and mammary tumors in Noble rats. Since this model of hormone-induced neoplasia has not been reported in other rat strains, we studied the effect of testosterone on the mammary gland morphology of female Wistar rats. Sixty adult, non-castrated, female Wistar rats were implanted in the dorsum midline with a silicone tube containing 50 mg testosterone (testosterone propionate in 30 animals and non-esterified testosterone in the remaining 30 animals) and 20 additional animals were implanted with empty tubes and used as control. Five animals per group were killed 30, 60, 90, 120, 150, and 180 days after implantation, and the mammary glands were dissected, fixed and embedded in paraffin. Histological sections were then stained with hematoxylin and eosin and picrosyrius red for collagen visualization. Morphological and morphometric analysis demonstrated ductal proliferation and acinotubular differentiation with secretory activity in all treated animals, peaking at 90 days of androgen exposure. After 90 days the proliferation of acinar epithelial cells was evident, but there was a progressive reduction of secretory differentiation and an increase in intralobular collagen fibers. There was no morphological evidence of dysplastic changes or other pre-neoplastic lesions. Testosterone treatment applied to adult, non-castrated female Wistar rats induced a mammary gland hyperplasia resembling the lactating differentiation, with progressive reduction in secretory differentiation.

Chronotropic responses to acetylcholine in atria of mice Chronically infected with Y and CL strains of Trypanososma cruzi

Cardiac parasympathetic denervation has been reported to occur in human experimental T. cruzi infections. We have used acetylcholine to investigate the in vitro chronotropic sensitivity of atrial muscarinic receptors in mice chronically infected with either a reticulotropic (Y) or a myotropic (CL) strain of T. cruzi. The dose-response curves did not from that obtained for control mice, a result that may be due to reinnervation processes