Metodologia de infecção experimental e grau de susceptibilidade do híbrido tambacu e Leporinus macrocephalus Garavello & Britski (Osteichthyes, Anostomidae) a quatro inóculos de trofozoítos de Icthyophthirius multifiliis Fouquet (Protozoa, Ciliophora)

Tropical fishes like Leporinus macrocephalus Garavello & Britski, 1988 (piauçu) and the hybrid tambacu (Piaractus mesopotamicus Holmberg, 1887 pacu male x Colossoma macropomum Cuvier, 1818 tambaqui female) were experimentally infected with four inocula of trophonts of Ichthyophthirius multifiliis Fouquet, 1876 (Protozoa, Ciliophora) and the infection level was evaluated. Each one of twelve experimental units consisted of a tank with 200 liters of water and two specimens of each kind of fish. The following treatments were designed to each tank, in triplicate: 8, 16,32 and 64 trophonts. After 8 days of exposure the fishes were killed and the number of trophonts was counted at a stereoscopic microscope. This assay showed to be feasible at tropical conditions and was able to show that tambacu was more susceptiblc than piauçu.

Infecção experimental por Salmonella enterica subespécie enterica sorotipo Panama e tentativa de transmissão nasonasal em leitões desmamados

O objetivo deste experimento foi produzir uma infecção experimental de Salmonella enterica subespécie. enterica sorotipo Panama e verificar a importância da via nasonasal na transmissão entre leitões desmamados. Foram utilizados seis leitões recém-desmamados, adquiridos de granja livre de Salmonella spp. Utilizaram-se baias isoladoras, que proporcionavam o contato nasonasal e eliminavam a possibilidade de outras vias de transmissão e de contaminação externa. Três grupos foram formados: controle, sentinela e infectado. Não foram encontradas amostras positivas para Salmonella spp. em leitões do grupo-controle e sentinelas, e nos animais infectados foi isolada Salmonella Panama em suabes retais e tecidos necropsiados. Os resultados revelaram não haver a transmissão pela via nasonasal entre leitões desmamados, pois, em nenhum momento, o agente foi isolado dos animais sentinelas

Natural killer cell activity in experimental paracoccidioidomycosis of the Syrian hamster

Com o objetivo de avaliar a atividade de células natural killer na paracoccidioidomicose experimental do hamster, 80 hamsters foram infectados por via intratesticular com Paracoccidioides brasiliensis e sacrificados após 24h, 48h, 96h, 1, 2, 4, 8 e 11 semanas de infecção. Como controle foram avaliados 40 hamsters normais, não infectados. Os animais foram submetidos ao estudo da atividade citotóxica de células NK pela técnica de single-cell assay e da resposta imune humoral pelas técnicas de imunodifusão dupla e Elisa. A produção do fator inibidor da migração de macrófagos em presença de Phytohemaglutinina e antígeno de P. brasiliensis e a histopatologia das lesões foram estudadas após 1,4, 8e 11 semanas de infecção. Os animais infectados, quando comparados aos controles, apresentaram níveis de atividade NK significativamente elevados durante as 4 primeiras semanas de infecção, havendo diminuição dessa atividade a partir da 8ª semana. Foi observada correlação inversa entre atividade NK e níveis de anticorpos específicos e, associação entre diminuição da atividade NK, depressão de resposta imune celular e aumento da extensão das lesões histopatológicas. Os resultados sugerem que após ativação inicial, as células NK são incapazes de controlar a disseminação do fungo pelos tecidos. A depressão da atividade NK na fase tardia da infecção parece estar relacionada com os distúrbios imunorregulatórios associados à paracoccidioidomicose.

Response of Corriedale and Crioula Lanada sheep to artificial primary infection with Haemonchus contortus

Clinical, parasitological and biochemical parameters were evaluated in Corriedale and Crioula Lanada sheep after a single experimental infection with Haemonchus contortus. Ten 4-month-old worm-free lambs, of each breed, were infected with 200 L-3 H. contortus per kg live weight,and four uninfected animals of each breed were used as controls. Every week, the animals were weighed and blood and faecal samples were collected for measurement of packed cell volume (PCV), total serum protein (TSP) and albumin (ALB), and the number of eggs per gram of faeces (EPG), respectively. Twelve weeks after infection, the animals were slaughtered. The worm burden was determined and samples of the abomasal mucosa were processed for determination of the number of eosinophils, mast cells and globule leukocytes. No significant differences in PCV, TSP, ALB, parasite burden or the cell populations of the abomasal mucosa were observed between breeds, but Crioula lambs had a lower EPG count. The comparison of the infected groups with their respective controls revealed significant alterations in PCV, TSP and ALB in the Corriedale lambs and in PCV, TSP, ALB and the density of eosinophils and mast cells in the Crioula lambs.

INTRAGASTRIC INFECTION OF CONVENTIONAL AND GERM-FREE MICE WITH GIARDIA-LAMBLIA

The effects of experimental infection with Giardia lamblia were studied in 30-day old conventional and germfree CFW mice (7 animals in each group) of both sexes. Cysts were observed in the feces of both groups 6 to 7 days after intragastric infection of each animal with about 2.5 x 10(5) G. lamblia trophozoites. Fecal cyst level was statistically higher in germfree mice (about 10(5) cysts/g feces) when compared with the conventional group (about 10(4) cysts/g feces). The peak of infection in the conventional group apparently occurred on the 10th day after infection as indicated by an increase of fecal weight and by histopathological examination. Intense infiltration of the lamina propria and high reactional hyperplasia of the lymphoid component were observed in the conventional group. There was no infiltration or hyperplasia in germfree infected mice and fecal weight was relatively constant throughout the experiment. These results suggest that, as is the case for other intestinal pathogenic protozoa, the intestinal microflora is indispensable for the expression of the pathogenicity but not for the multiplication of G. lamblia.

EARLY ACTIVATION OF SPLENIC MACROPHAGES BY TUMOR-NECROSIS-FACTOR-ALPHA IS IMPORTANT IN DETERMINING THE OUTCOME OF EXPERIMENTAL HISTOPLASMOSIS IN MICE

Experimental infection of animals with Histoplasma capsulatum caused a massive macrophage infiltration into the spleen and induced the production of tumor necrosis factor alpha (TNF-alpha) locally. The cytokine was also produced in vitro by peritoneal exudate macrophages exposed to a large inoculum of yeast cells. Depletion of the cytokine by injection of polyclonal sheep anti-TNF-alpha antibody was detrimental to sublethally infected mice. Fungous burdens in the spleens of TNF-alpha-depleted mice were higher than they were in the infected control mice at days 2, 7, and 9 after infection, and the antibody-treated animals succumbed to the infection. Histopathological study of spleen sections revealed that splenic macrophages were not able to control proliferation of intracellular yeasts as a result of TNF-alpha depletion. It seems that TNF-alpha plays a role in early activation of splenic macrophages which is important in controlling the outcome of an infection.

Efficacy of florfenicol and intravenous fluid therapy for treatment of experimental salmonellosis in newborn calves

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Apoptotic and developmental effects of bovine Herpesvirus type-5 infection on in vitro-produced bovine embryos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Bull semen varibles after experimental exposure with Bovine Herpesvirus type 5

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The effect of Ketoconazole on experimental paracoccidioidomycosis in the Syrian hamster: immunological and histopathological study

The effect of Ketoconazole (KTZ) on the hamster experimental intratesticular paracoccidioidomycosis was studied employing different treatment schedules. KTZ long course treatment beginning at an early stage of the infection was effective in preventing fungal proliferation, dissemination to lymph nodes, spleen and kidneys, and in maintaining low levels of humoral and cellular specific immune responses. KTZ short course treatment starting at an advanced stage of disease resulted in a more severe histopathological picture without significant changes in the immunological profile. The drug prolonged the life span of hamsters infected with Paracoccidioides brasiliensis, but did not prevent mortality. Toxic necrosis of the bone marrow occurred in normal animals receiving 120 mg/kg/day of KTZ but with lower doses no morphologic alterations were observed in heart, lungs, kidneys, adrenals, spleen, liver, intestine or bone marrow. © 1984 Dr W. Junk Publishers.

Ketoconazole in the treatment of experimental murine paracoccidioidomycosis

In a murine model of chronic disseminated paracoccidioidomycosis (strain 18; intravenous route), Ketoconazole (200 mg/kg in 0.2% agar) was given daily by gavage in three different schedules. Continuous treatment from an early stage of infection (day 3) up to week 20 was the most effective protocol, leading to remission of histopathological lesions and of both humoral and cellular anti-P. brasiliensis immune response, and clearance of the fungus in lungs; only 1 treated animal at week 20 showed pulmonary granulomas, although less extensive than control mice. Continuous treatment from early stage up to week 8, followed by a 16 week-period of drug discontinuity, caused remission of lesions in all but 3 treated mice which showed active pulmonary paracoccidioidomycosis similar to controls (14.2% of unresponsiveness to treatment). The continuous Ketoconazole protocol since a late stage of infection (week 4) up to week 20 produced a slower remission of lesions and immune response when compared with the first drug schedule. In this model of paracoccidioidomycosis, Ketoconazole showed no detectable side-effects and was a very effective drug especially in a prolonged administration protocol from an early stage of infection.

A novel biological activity for galectin-1: Inhibition of leukocyte-endothelial cell interactions in experimental inflammation

Galectin-1 (Gal-1), the prototype of a family of β -galactoside-binding proteins, has been shown to attenuate experimental acute and chronic inflammation. In view of the fact that endothelial cells (ECs), but not human polymorphonuclear leukocytes (PMNs), expressed Gal-1 we tested here the hypothesis that the protein could modulate leukocyte-EC interaction in inflammatory settings. In vitro, human recombinant (hr) Gal-1 inhibited PMN chemotaxis and trans-endothelial migration. These actions were specific as they were absent if Gal-1 was boiled or blocked by neutralizing antiserum. In vivo, hrGal-1 (optimum effect at 0.3 μg equivalent to 20 pmol) inhibited interleukin-1β-induced PMN recruitment into the mouse peritoneal cavity. Intravital microscopy analysis showed that leukocyte flux, but not their rolling velocity, was decreased by an anti-inflammatory dose of hrGal-1. Binding of biotinylated Gal-1 to resting and post-adherent human PMNs occurred at concentrations inhibitory in the chemotaxis and transmigration assays. In addition, the pattern of Gal-1 binding was differentially modulated by PMN or EC activation. In conclusion, these data suggest the existence of a previously unrecognized function of Gal-1, that is inhibition of leukocyte rolling and extravasation in experimental inflammation. It is possible that endogenous Gal-1 may be part of a novel anti-inflammatory loop in which the endothelium is the source of the protein and the migrating PMNs the target for its anti-inflammatory action.

Relevance of Chinese goose (Anser cygnoides) in experimental epidemiology of Newcastle disease

This study aimed to characterize the true epidemiological role played by the Chinese goose (Anser cygnoides) as a potential source of infection by the Newcastle disease virus (NDV). For this, Specific-Pathogen-Free chicks (SPF) were used and were housed with Chinese geese that had been inoculated with a pathogenic strain (velogenic viscerotropic, strain São João do Meriti) of NDV (DIE50=108.15/0.1 mL) pathogenic to chickens, by the ocular-nasal route. Each group was composed of 6 SPF Leghorn chicks and 3 geese. At 6 days (Group I) and 14 days (Group II) after inoculation of the Chinese geese with NDV, SPF chicks were put into direct contact with each goose group. Cloacal swabs were collected from both species (Chinese geese and SPF chicks) 6, 10 and 20 days after challenge to genome viral excretion by Reverse Transcription Polymerase Chain Reaction (RT-PCR). Chinese geese did not demonstrate any clinical signs of Newcastle disease (ND). They were refractory to the clinical disease with the NDV. However, NDV genome was detected 20 days after challenge. Therefore, NDV carrier status was demonstrated by Chinese geese. Moreover, 100% of SPF chicks housed with the infected Chinese geese had died by 6 (Group I) and 14 days (Group II) after challenge. Thus, the transmission of the pathogenic virus from the Chinese geese to cohabiting SPF chicks was evident within 20 days of the experimental infection. This reveals the epidemiological importance of Chinese geese as a potential transmitter of NDV infection to other commercial birds that could be raised in close proximity.