Avaliação clínico-laboratorial de bovinos Nelore infectados experimentalmente com Trypanosoma vivax

Avaliaram-se as alterações clínico-laboratoriais de seis bezerros Nelore, de ambos os sexos, inoculados experimentalmente com 10(7) organismos viáveis de Trypanosoma vivax, isolados de bovinos da região de Poconé, Estado de Mato Grosso. Os animais foram observados diariamente, durante 30 dias, quanto aos parâmetros de temperatura retal, volume globular (VG), parasitemia, produção de anticorpos, coloração de mucosas, comportamento e apetite. Determinaram-se os níveis séricos de aspartato aminotransferase (AST), fosfatase alcalina (FA), gama glutamiltransferase (GGT), creatina kinase (CK), colesterol, uréia, creatinina, cálcio, fósforo e o perfil eletroforético das proteínas séricas aos 4, 8, 12, 16, 23 e 30 dias pós-inoculação (DPI). Durante os 6 meses seguintes, os animais foram observados semanalmente, avaliando-se a temperatura retal, o VG e a parasitemia. T. vivax foi evidenciado a partir do terceiro e quarto DPI em todos os bezerros e persistiu até o 30° DPI em cinco dos seis animais em estudo. Ocorreu um decréscimo significativo (p<0,05) do valor médio do VG (25%) aos dez DPI. Os animais não apresentaram qualquer alteração no quadro clínico, bem como na avaliação da bioquímica sérica durante o período experimental. A soroconversão ocorreu aos 6 e 8 DPI, permanecendo todos os animais soropositivos nos 30 dias experimentais. Bovinos nelores jovens, infectados experimentalmente com T. vivax, foram capazes de estabelecer um equilíbrio na relação hospedeiro-parasita.

Partial characterization of proteolytic activity in Giardia duodenalis purified proteins

O presente estudo consiste em uma caracterização preliminar da atividade proteolítica de frações de proteínas purificadas a partir de lisados de trofozoítos de cepa isolada e axenizada no Brasil. Frações obtidas por cromatografia líquida (FPLC) foram analisadas quanto ao perfil eletroforético em géis de poliacrilamida (SDS-PAGE) e a atividade proteolítica foi avaliada em géis contendo gelatina como substrato. A caracterização das enzimas foi realizada a partir da análise do efeito de inibidores sintéticos de cisteína-proteases (E-64, IAA), serina-proteases (PMSF), serina e cisteína-proteases (TPCK, TLCK, elastatinal), metalo-proteases (EDTA) e aspartil proteases (pepstatina) sobre a degradação do substrato. Entre 30 frações eluídas, bandas de proteínas foram observadas em oito delas, entretanto, atividade proteolítica foi detectada apenas nas frações 23, 24, 25 e 26. O perfil eletroforético das proteínas revelou poucas bandas distribuídas na faixa de 45 a 18 kDa. Os zimogramas revelaram zonas de proteólise na faixa de aproximadamente 62 a 35 kDa, entretanto destacaram-se as bandas de hidrólise de 62, 55, 53, 50, 46 e 40 kDa. Nos ensaios de inibição, a proteólise foi marcantemente inibida por E-64, TPCK, TLCK e elastatinal. Redução discreta da proteólise foi observada com IAA e PMSF, enquanto que EDTA e pepstatina não promoveram alteração dos perfis de hidrólise. Estas observações são relevantes, especialmente se considerarmos que para elucidar o envolvimento das proteases na relação parasita-hospedeiro, a purificação dessas moléculas é um requisito importante.

Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoites

One of the major questions concerning Giardia is the understanding of pathophysiological processes associated with small intestine abnormalities. There are evidences that Giardia trophozoites contain and/or release proteolytic enzymes that may be implicated in the host intestinal epithelium. The present investigation was undertaken to examine the protease activity in excretory/secretory (E/S) products of Giardia duodenalis trophozoites of an axenic Brazilian strain (BTU-11) and the reference strain Portland 1 (P1). E/S products from trophozoites of each strain in conditioned medium were tested with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for the protein profiles, and the protease activity was analyzed using substrate-impregnated SDS-PAGE (gelatin and collagen) and hemoglobin assay. The proteases characterization was based on inhibition assays including synthetic inhibitors. Electrophoresis analysis of E/S products revealed a banding pattern composed by few bands (4 to 6 bands) in the migration region of 123 to 28 kDa. Proteolytic products were detected in the conditioned medium by trophozoites of both assayed strains. In the gels containing copolymerized gelatin and collagen, E/S products promoted substrate degradation and the most evident proteolysis zones were distributed in the migration regions of 77 to 18 kDa and 145 to 18 kDa, respectively, in the patterns of gelatinolytic and collagenolytic activities. Degradation of hemoglobin was also observed, and the pattern of hydrolysis was similar in both E/S products assayed. Inhibitor assays showed that the main proteolytic activity in both E/S products is due to cysteine proteases, although the presence of serine proteases was also indicated. Degradation of substrates including collagen and hemoglobin could lead us to speculate different functions of Giardia excreted/secreted proteases in vivo, but to confirm this possibility and to elucidate its implication on host-parasite interactions, further experiments applying protocols for the purification of proteases are necessary. Even so, our observations are relevant and hold the perspective for the understanding about protease activity in Giardia trophozoites of axenic strain isolated in an endemic area.

Immunity in plants and animals: common ends through different means using similar tools

A comparative approach is potentially useful for understanding the role of mammal innate immunity role in stimulating adaptive immunity as well as the relationship between these two types of immune strategies. Considerable progress has been made in the elucidation of the co-ordinated events involved in plant perception of infection and their mobilisation of defence responses. Although lacking immunoglobulin molecules, circulating cells, and phagocytic processes, plants successfully use pre-formed physical and chemical innate defences, as well as inducible adaptive immune strategies. In the present paper, we review some shared and divergent immune aspects present in both animals and plants. (C) 2002 Elsevier B.V. All rights reserved.

Production, purification, crystallization and preliminary X-ray diffraction studies of the nucleoside diphosphate kinase b from Leishmania major

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

Experimental paracoccidioidomycosis in hamster: transmission electron microscopy of inoculation site lesion

Estudou-se sequencialmente, à microscopia eletrônica de transmissão, a interação entre Paracoccidioides brasiliensis (Pb) e células inflamatórias em hamsters inoculados por via intratesticular. Seis horas após inoculações havia predominância de neutrófilos, estando presentes algumas células mononucleares e eosinófilos. Os neutrófilos foram progressivamente substituídos por células mononucleares. Fungos viáveis apresentavam-se fagocitados ou circunscritos por células inflamatórias, geralmente com ampla interface hospedeiro-parasita. Fungos mortos ou degenerados eram acompanhados de interfase estreita. A camada externa da parede do Pb era às vezes quebrada quando em contacto com neutrófilos, em vários pontos, sendo os fragmentos dessa parede descamados e fagocitados. Células fúngicas pequenas com um único núcleo se relacionavam com ampla interfase enquanto as células maiores e multinucleadas apresentavam paredes irregulares, por vezes, contendo lomasoma e/ou estrutura semelhante à mielina. Diferentes padrões de interação do Pb com células do hospedeiro podem ser decorrentes do a fluxo de células inflamatórias funcionalmente diferentes ao local de inoculação ou à idade dos fungos ou ambos os fatores.

DISTRIBUTION OF THE NUMBER OF EURYTREMA SP EGGS PER GRAM OF FECES IN NATURALLY INFECTED CATTLE

Eurytrema sp. egg counts (epg) in the feces of naturally infected cattle were performed and the technique employed showed 94.2% probability of detecting positive cases of the infection with a single examination independently of the host parasite burden. It was also demonstrated that the epg of Eurytrema sp. follows a negative binomial distribution model and is characterized by its small magnitude.

Effects of sex and locality on the abundance of lice on the wild rodent Oligoryzomys nigripes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Inhibitory effect of PGE(2) on the killing of Paracoccidioides brasiliensis by human monocytes can be reversed by cellular activation with cytokines

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

Similarities among ectoparasite fauna of sigmodontine rodents: phylogenetic and geographical influences

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Implicações da agregação espacial de parasitas para a dinâmica populacional na interação hospedeiro-parasita

Some aspects of the widely observed over-dispersed pattern of the distribution of parasites within the host population are examined. It has been established in the parasitological literature that most hosts usually harbour few parasites, while only few hosts harbour a large proportion of the parasite population. Factors that may influence the pattern of distribution of parasites, the relation between the level of parasite aggregation and the prevalence of infection, and changes in this level of aggregation as a function of host age are analysed. Factors which determine the diversity of species in parasite communities are presented, and aspects of exploitative and interference competition among parasites and their relations with biological control procedures are also considered. Attention is also focused on the regulatory and destabilizing processes influencing the dynamic behaviour of host-parasite population interactions.

Invited review Giardia duodenalis: Inter-strain variability of proteins, antigens, proteases, isoenzymes and nucleic acids

Giardia duodenalis isolates from asymptomatic or symptomatic patients and from animals present similarities and differences in the protein composition, antigenic profile, pattern of proteases and isoenzymes, as well as in nucleic acids analysis. In the present overview, these differences and similarities are reviewed with emphasis in the host-parasite interplay and possible mechanisms of virulence of the protozoon.

Ticks (Acari: Ixodidae) on Wild Marsh-Deer (Blastocerus dichotomus) from Southeast Brazil: Infestations before and after Habitat Loss

The lake from Porto-Primavera hydroelectric power station inundated an area of 2,200 km2 at the border of São Paulo and Mato-Grosso do Sul States, Brazil. Infestations by ticks were evaluated on 135 marsh deer, Blastocerus dichotomus (Illiger), captured before and after inundation. Ticks were collected for identification, and infestation level of animals was assessed by scoring. Deer were divided into four groups according to capture location and temporal relation to the inundation. Groups 1, 2, and 3 were captured before inundation. Group 4 was captured after inundation. Four tick species were found: Amblyomma cajennense (F.), Amblyomma triste Koch, Anocentor nitens (Neumann), and Boophilus microplus (Canestrini). Groups 1, 2, 3, and 4 had 30, 45, 100, and 96%, respectively, of animals carrying B. microplus ticks. A. triste was observed on 16, 22, 22, and 88% of animals from groups 1,2,3, and 4, respectively. A. nitens and A. cajennense were observed only on group 4, on 32 and 16% of the animals, respectively. Groups 1 and 2 had only 4.8 and 6.1% of animals with high infestation levels, respectively, and no ticks on 46.8% and 45.5% of the animals, respectively. Conversely, groups 3 and 4 lacked noninfested animals and had high infestation levels on 77.8 and 50% of deer, respectively. Marsh area shrinkage was blamed for higher infestation levels on deer from groups 3 and 4. The widespread presence of A. triste on marsh deer, a Neotropical tick species, raises the possibility of a natural host-parasite relationship.

Antigens from Rhipicephalus sanguineus ticks elicit potent cell-mediated immune responses in resistant but not in susceptible animals

In the present study we compared the immunological reactions between Rhipicephalus sanguineus tick-infested susceptible (dogs and mice) and tick-resistant hosts (guinea pigs), elucidating some of the components of efficient protective responses against ticks. We found that T-cells from guinea pigs infested with adult ticks proliferate vigorously in the presence of concanavalin A (ConA), whereas ConA-induced cell proliferation of tick-infested mice and dogs was significantly decreased at 43.1 and 94.0%, respectively, compared to non-infested controls. Moreover, cells from mice and dogs submitted to one or three successive infestations did not exhibit a T-cell proliferative response to tick antigens, whilst cells from thrice tick-infested guinea pigs, when cultured with either a tick extract or tick saliva, displayed a significant increase in cell proliferation. Also, we evaluated the response of tick-infested mice to a cutaneous hypersensitivity test induced by a tick extract. Tick-infested mice developed a significant immediate reaction, whereby a 29.9% increase in the footpad thickness was observed. No delayed-type hypersensitivity (DTH) reaction was detected. Finally, the differential cell count at the tick attachment site in repeatedly infested mice exhibited a 6.6- and 4.1-fold increase in the percentage of eosinophils and neutrophils, respectively, compared to non-infested animals, while a decrease of 77.0-40.9 in the percentage of mononuclear cells was observed. The results of the cutaneous hypersensitivity test and the cellular counts at the tick feeding site for mice support the view that tick-infested mice develop an immune response to R. sanguineus ticks very similar to dogs, the natural host of this species of tick, but very different from guinea pigs (resistant host), which develop a DTH reaction in addition to a basophil and mononuclear cell infiltration at the tick-attachment site. In conclusion, saliva introduced during tick infestations reduces the ability of a susceptible animal host to respond to tick antigens that could stimulate a protective immune response. As a consequence, the animals present a lack of DTH response and disturbed cellular migration to tick feeding site, which can represent a deficient response against ticks. © 2003 Elsevier B.V. All rights reserved.

Fauna parasitária de peixes oriundos de pesque-pague do município de Guariba, São Paulo, Brasil

This study describes the occurrence and the seasonality of parasites of cultivated fish from a fee fishing farm located in Guariba, São Paulo State, Brazil (21°15'22'' S, 48°18'58'' W and 595 m of altitude), from August, 2001 to July, 2002. The presence of parasites was researched in pacu Piaractus mesopotamicus (Characidae), common carp Cyprinus carpio (Cyprinidae), nile-tilapia Oreochromis niloticus (Cichlidae), tambacu hybrid (male of P. mesopotamicus x female of tambaqui-Colossoma macropomum) and piraputanga Brycon hillari (Characidae). Results demonstrate that out of 100 fish examined, 15% were sponged for at least one of the following parasites: Trichodina sp.; monogenean helminths; copepodits of Lemaea cyprinacea; adults of L. cyprinacea; or Dolops carvalhoi. In decreasing order, the susceptibility degree of the hosts was C. carpio, P. mesopotamicus, B. hillari, tambacu hybrid and O. niloticus. In decreasing order the reported parasites were monogenean helminths, Dolops carvalhoi, Trichodina sp., adults of Lernaea cyprinacea and their young shapes.