Estimativa da concentração espermática do sêmen de peixe pelo método de espermatócrito

Um experimento foi realizado com o objetivo de estimar a concentração espermática das espécies dourado (Salminus brasiliensis), curimba (Prochilodus lineatus), jundiá (Rhamdia quelen), cascudo-preto (Rhinelepis aspera) e tilápia-do-nilo (Oreochromis niloticus) pelo método de espermatócrito. Utilizaram-se 19, 58, 51, 43 e 85 reprodutores de dourado, curimba, jundiá, cascudo-preto e tilápia-do-nilo, respectivamente. Com exceção da tilápia-do-nilo, os reprodutores foram submetidos ao processo de indução hormonal e posteriormente submetidos a coleta de sêmen. Foram comparadas as técnicas de mensuração da concentração espermática do sêmen por contagem em câmara hematimétrica de Neubauer e por espermatócrito. Os resultados obtidos foram submetidos à análise de regressão a 5% de probabilidade. As concentrações espermáticas mensuradas por ambas as técnicas apresentaram relação linear, para curimbas, jundiás e tilápias-do-nilo, com equações y = 6,6624 × 10(9) + 3,68553 × 10(8)x; y = 2,153 × 10(9) + 4,426 × 10(8)x e y = -9,0897 × 10(8) + 6,0167 × 10(8), respectivamente. O método de espermatócrito pode ser utilizado para estimar a concentração espermática do sêmen de curimbas, jundiás e tilápias-do-nilo.

Study of some Brazilian Myrtaceae in Jaboticabal sp

The family Myrtaceae is one of the most important related to fruit species. In Brazil it encloses more than 100 species of native fruit from North to South of the country (Giacometti, 1992; Donadio, 1992; Mattos, 1992). The genus Eugenia, Campomanesia, Psidium and Myrciaria are the most important, grouping most of the species of some importance. The germplasm bank of the Universidade Estadual Paulista includes some of the major species. Some characteristics are typical for most of the Myrtaceae fruit species. The most relevant are the long juvenile period presented by the trees, taking long time to produce, when propagated by seed; the slow growing tree behavior; difficult to propagated by vegetative means; large variability in trees obtained by seed propagation. The cited characteristics are commented for some species as well as the fruit quality, from data obtained in trees at the same ecological conditions.The species evaluated are: 1) Eugenia stipitata Mc Vaugh; 2) Eugenia tomentosa Cambes; 3) Eugenia brasiliensis Lam.; 4) Eugenia uniflora L.; 5) Eugenia luchsnathiana Berg.; 6) Eugenia uvalha Cambess; 7) Camponesia spp; 8) Plinia edulis (Veil. Sobral); 9) Eugenia involucrata D.C.; 10) Psidium acutangulum Mart.; 11) Myrciaria dubia Mc Vaugh; 12) Engenia guabiju Berg. The main conclusions based on the data analysed are: the fruits of some species present good general characteristics, mainly for fresh consumption, as high Vitamin C, sugars, pulp and other characteristics; 'Araca-boi' presents some good fruit characteristics, but is more adapted for processing, by its high acidity; Some species present low % of edible portion and need to be improved to gain commercial acceptance. mainly the 'cabeludinha' and 'cambuca'.

Differences in reactivity of paracoccidioidomycosis sera with gp43 isoforms

The glycoprotein gp43 from Paracoccidioides brasiliensis is the main antigenic component in paracoccidioidomycosis (PCM) because it is recognized by 100% of PCM patients. It has also been shown that different fungal strains produce gp43 with at least four isoform profiles. In this study, different isoform profiles from gp43, with pIs ranging from 5.8 to 8.5, were affinity purified from various P. brasiliensis (B-339, S.S., 1925 and I-9) exoantigens. Because of the isoform heterogeneity, we questioned whether those isoform profiles could be similarly recognized by acute or chronic PCM patients. By using a specific and sensitive method for detection of human IgG anti-gp43 antibodies, the monoclonal antibody capture immunoassay, we report that not all gp43 isoform profiles are equally recognized in PCM sera when anti-gp43 MAb 17c was employed as capturing antibody. Our result showed that recognition of pI 8.5 gp43 isoform was significantly lower for both acute (56%) and chronic patients (71%), compared with gp43 isoforms from the standard strain B-339. on the other hand, when anti-gp43 MAb 8a, which recognizes a different antigenic epitope was used to capture the different gp43 isoform profiles, all patient's sera reacted similarly. The results described suggest that not all the antigenic epitopes expressed by gp43 are equally present in all P. brasiliensis strains.

Chloroquine is therapeutic in murine experimental model of paracoccidioidomycosis

Chloroquine, due to its basic properties, has been shown to prevent the release of iron from holotransferrin, thereby interfering with normal iron metabolism in a variety of cell types. We have studied the effects of chloroquine on the evolution of experimental paracoccidioidomycosis by evaluating the viable fungal recovery from lung, liver and spleen from infected mice and H2O2, NO production, tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-6, IL-10 levels and transferrin receptor (TfR) expression from uninfected and infected peritoneal macrophages. Chloroquine caused a significant decrease in the viable fungal recovery from all organs tested, during all periods of evaluation. Peritoneal macrophages from chloroquine-treated infected mice showed higher H2O2 production and TfR expression, and decreased levels of NO, endogenous and stimulated-TNF-alpha, IL-6 and IL-10 during the three evaluated periods. However, despite its suppressor effects on the macrophage function, the chloroquine therapeutic effect upon murine paracoccidioidomycosis was probably due to its effect on iron metabolism, blocking iron uptake by cells, and consequently restricting iron to fungus growth and survival.

EXPERIMENTAL PARACOCCIDIOIDOMYCOSIS OF HAMSTER INOCULATED IN THE CHEEK POUCH

We compared the granuloma morphology and immune response of hamsters inoculated with Paracoccidioides brasiliensis (Pb) into the cheek pouch, which lacks lymphatic drainage, and into the footpad, which is rich in lymphatics. Our objective was to better understand the modulation of Pb granuloma in an immunocompetent animal inoculated in an immunologically privileged site. The humoral immune response (ELISA) and cell mediated immunity (footpad test) became positive on days 7 and 14, respectively in animals inoculated into footpad and on days 35 and 60 in animals inoculated into the pouch. Typical epithelioid granulomas were observed at both sites on day 14. The number of fungi gradually decreased from the beginning of the experiment in footpad lesions, but only after day 35 in pouch granulomas, when cell mediated immunity was detectable. The results indicate that typical epithelioid paracoccidioidomycotic granulomas may develop in the absence of a detectable immune response; however, they are incapable of controlling fungal reproduction. Lack of lymphatic drainage delays the appearance of a detectable immune response, but with time fungi escape from the pouch, elicit an immune response and reach other organs. Our results further indicate the importance of the lymphatics in the pathogenesis of paracoccidioidomycosis.

Pattern of nucleolar activity during spermiogenesis in triatomines (Heteroptera, Reduviidae) as analysed by silver staining

Nucleoli are the sites of biosynthesis of ribosomal precursors. In this work the nucleolar activity at interphase and the meiotic cells of the testis in five species of triatomines were analysed by means of silver staining. Several nucleolar blocks in the polyploid nuclei of testicular tubules were observed, whereas only one nucleolar body could be seen in the spermatogonial nuclei of all five species. A single nucleolar body was evident in the 'confused stage' of Triatoma brasiliensis, T. delpontei, T. lecticularia and T. rubrovaria, while T. sordida presented two nucleolar dots. The existence of small, silver-stained dots in some metaphase I chromosomes of T. brasiliensis and T. sordida is reported. The number of nucleolar dots present in spermatids of each species varied within and among species. It is suggested that in addition to providing information on rRNA biosynthesis, studies of nucleolar organizing activity can also be important sources of data on differentiation patterns and species development.

Correlation among immune response, morphogenesis of the granulomatous reaction and spleen lymphoid structure in murine experimental paracoccidioidomycosis

We studied the correlation among cellular immune response, the pattern of lung granulomatous lesions and alterations in spleen lymphoid structure in Swiss mice inoculated intravenously with Paracoccidioides brasiliensis strain 18. The animals were evaluated at 24, 48 and 96 h after infection and further studied weekly for 18 weeks by: (i) the macrophage migration inhibition test with phytohemagglutinin (PHA) and P. brasiliensis antigen (PbAg); and (ii) histopathology of the lung and spleen lesions. One group of animals was gamma -irradiated (8 Gy), infected under the same conditions and evaluated for the pattern of lung granulomatous lesions and spleen lymphoid structure at 24, 48 and 96 h after infection. During the first week of infection, the non-irradiated animals presented a positive response to PHA and PbAg, compact granulomas in the lungs and a typical hyperplasia of the spleen white pulp. However, from weeks 2 to 5, a depression of the cell-mediated immunity (CMI) response to PHA and PbAg was observed in association with granulomas presenting only large mononuclear cells and lacking both giant cells and a peripheral halo of small mononuclear cells. This pattern of granuloma formation was similar to that seen in gamma -irradiated animals, whose cells involved in CMI were absent. After week 7, the non-irradiated animals showed granulomas characterized by the presence of giant cells and a peripheral halo of small mononuclear cells. This type of granuloma was formed concomitantly with recovery of the CMI and of the lymphoid structure of the spleen. The results showed a correlation among granulomas composed of large mononuclear cells, hypoplasia of the splenic tissue and impaired CMI. This correlation indicated that although granuloma morphogenesis per se does not depend on the activation of CMI, this response is important at later stages during modulation of the cellular composition of the granulomas.

Ribosomal DNA ITS-1 intergenic spacer polymorphism in triatomines (Triatominae, Heteroptera)

The length polymorphism of ribosomal DNA ITS-1 intergenic spacer was analyzed in eight species of triatomines belonging to Triatoma, Rhodnius, and Panstrongylus genera. The analyzed species were Rhodnius domesticus, R. neivai, R. robustus, Triatoma brasiliensis, T. infestans, T. vitticeps, Panstrongylus megistus, and P. herreri. These insects are vectors of Chagas' disease, one of the most prominent public health problems among South American countries. This work allowed the differentiation between species of the Triatomini and Rhodniini tribes through the analysis of ITS-1 length polymorphism by PCR and RFLP techniques. The species of the Triatoma and Panstrongylus genera presented an amplified ITS-1 fragment between 600 and 1000 bp, whereas Rhodnius presented a less variable ITS-1 length fragment, around 300 bp, which could reflect the monophyletic origin of the Rhodniini tribe. Species belonging to this genus were further differentiated by RFLP with HaeIII and AluI endonucleases. Our results corroborate the hypothesis of polyphyletic origin in this group of insects and contribute to knowledge about evolutionary relationships in triatomines.

Spatial patterns or trees in the Picinguara coastal sandy forest (state of Sao-Paulo, Brazil)

The species of the sandy plains forests (forests of the ''restingas'') have not yet had their spatial patterns studied as aids to the understanding of the diversity found in the different physiognomies along the Brazilian coast. In this paper a 10 x 10 m quadrat framework laid in a hectare of a tree dominant forest in the sandy plains of the Picinguaba area of the Serra do Mar State Park (municipality of Ubatuba, state of São Paulo, Brazil) was used to assess the spatial pattern of distribution for the ten most important species : Pera glabrata, Euterpe edulis, Eugenia brasiliensis, Alchornea triplinervea, Guatteria australis, Myrcia racemosa, Jacaranda semiserrata, Guarea macrophylla, Euplassa cantareirae and Nectandra oppositifolia. The spatial patterns were inferred through the calculations of their T-Square Index (C) and Dispersal Distance Index (I). P. glabrata shows a random pattern, E. edulis aggregate, E. brasiliensis, A. triplinervia, G. australis, E. cantareirae and N. oppositifolia with a tendency between aggregate and uniform and, M. racemosa, J. semiserrata and G. macrophylla between aggregate and random. Although the indexes are dependent of the sample size and of the technique adjustments, the relationship of the pattern with the environmental factors is shown by clustering methods. The results give confirmation of how the spatial patterns bring associations between populations and shape of the vegetation physiognomy.

Study of pulmonary experimental paracoccidioidomycosis by analysis of bronchoalveolar lavage cells: Resistant vs. susceptible mice

Adult Swiss (susceptible) and BALB/c (non-susceptible) mice were inoculated by the intravenous route with 1 x 10(6) yeast cells of Paracoccidioides brasiliensis, strain 18. Immunologic parameters, histopathology and features of the bronchoalveolar lavage (BAL) were evaluated at week 2, 4, 8 and 16 post-infection. The pulmonary infection was progressive in Swiss mice and regressive in BALB/c mice. The numbers of total cells, lymphocytes and polymorphonuclear neutrophils increased in BAL, as well as the percentages of giant cells, and CD4 and CD8 positive cells. The ultrastructural study of BAL cells revealed a predominance of macrophages and a frequency of 13.2% of type II pneumocytes. As the infection progressed, the number of fungal cells and spreading macrophages, as well as the stimulated release of H2O2 by macrophages, increased. The animals exhibited an exacerbation of the humoral immune response and a depression of cellular immunity during the infection. There was a good correlation between the intensity and the pattern of the pulmonary histopathology and the cellular findings in the BAL. The present model reproduces some anatomoclinical patterns of the human disease and shows that BAL may be a useful tool in monitoring the pulmonary infection caused by P. brasiliensis.

Ocorrência e flutuação populacional de ácaros associados a seringais vizinhos de fragmentos de cerrado

The basic knowledge of the seasonal occurrence of mites can supply data for elaboration of programs of ecological management to be implanted with success in the future. The objective of this study was to determine the species richness and the seasonality of mites present in two areas of rubber tree crops neighboring to native areas in Itiquira, MT. Along one year, 25 quantitative samplings were accomplished in rubber tree crop neighboring to two fragments of Cerrado (Cerradao and Mata Riparia). There were registered 199,380 mites, of 48 species, belonging to 15 families. of those, 13 species are phytophagous, 18 predators and 17 mycophagous or of unknown alimentary habit. Three phytophagous species represented more than 97% of the mites collected: Phyllocoptruta seringueirae Feres (80.8%), Tenuipalpus heveae Baker (12.7%) and Calacarus heveae Feres (3.6%). Among the predators, the most abundant species were Agistemus sp., Scirula sp. and Euseius concordis (Chant). Twenty-eight species were common to both crops. The families that had the largest number of species collected in the neighboring area to Cerradao were Tydeidae (7), Tarsonemidae (6), Eriophyidae and Phytoseiidae (4), and in the area close to Mata Riparia, Tydeidae (9) and Phytoseiidae (8). The presence of vegetation near the crop should explain the great number of species of mites classified as accidental found in this study. The largest abundances and species richness occurred in the end of the rainy season and beginning of the dry season.

Cytotaxonomic analysis of Brazilian species of the genus Amazona (Psittacidae, Aves) and confirmation of the genus Salvatoria (Ribeiro, 1920)

The karyotypes of 12 species of Psittacidae of the genus Amazona were studied: A. aestiva, A. amazonica, A. brasiliensis, A. autumnalis, A. farinosa, A, festiva, A. kawalli, A. ochrocephala, A. pretrei, A. rhodocorytha, A. vinacea and A. xanthops. The metaphases were obtained using a short term culture of leather pulp. Eleven of the twelve analyzed species were karyotypically homogeneous, with only a few divergences in chromosomes 2 and 3. The species A. xanthops showed large karyotypic differences compared to the genus Amazona. Consequently, the genus Salvatoria (Ribeiro, Rev. Mus. Paul. 12: 1-82, 1920) was confirmed, and A. xanthops renamed Salvatoria xanthops. The study showed the chromosomic conservation of the genus Amazona and the need for further taxonomic studies of the karyotype of the Psittacidae.

Dialysable leukocyte extracts modify the course of experimental paracoccidioidomycosis in the Syrian hamster

The effect of dialysable leukocyte extracts (DLE) obtained from hamsters immunized with Paracoccidioides brasiliensis (immune DLE) and from non-immunized hamsters (non-immune DLE) was studied in hamsters inoculated with P. brasiliensis by the intratesticular route. Treatment with immune or non-immune DLE was started during the third week of infection and was repeated at 7, 11, 15 and 19 weeks. A group of untreated infected animals was used as control. Animals were submitted to the delayed hypersensitivity skin test to P. brasiliensis antigen (PbAg) in vivo and assayed in vitro by the macrophage migration inhibition test in the presence of Phytohemagglutinin (PHA) and PbAg and by immunodiffusion for specific antibody. The animals were sacrificed at 4, 8, 12, 16 and 20 weeks. The morphology and extension of the lesions were studied at the inoculation site, and in lymph nodes, lungs, liver, spleen and kidneys. In contrast to the controls, animals treated with both DLEs maintained a positive cell-mediated immune response throughout the experiment and developed less extensive infection with a significantly lower number of fungi in the lesions. The results suggest that immune and non-immune DLE preparations modified the evolution of experimental paracoccidioidomycosis with equal efficiency. This similarity may be explained by the immunoregulatory activities of both extracts.

SEXUAL DISTINCTION BETWEEN 5TH INSTAR NYMPHS OF 6 SPECIES OF TRIATOMINAE (HEMIPTERA, REDUVIIDAE)

The aim of this paper is to present the analysis of sexual morphological differences observed in 5th instar nymphs of the following species: Panstrongylus megistus; Rhodnius neglectus; Triatoma brasiliensis; T. infestans; T. matogrossensis and T. tibiamaculata. Male and female nymphs were examined and photographed with a Scanning Electron Microscope. The 9th segment dimensions of dorsal and ventral faces were determined through a Profile Projector. Results and statistical analysis showed significant differences: the 9th sternite is significantly broader in male than in female nymphs, while in five species, tergites in female nymphs are broad and in male are narrow.

Experimental paracoccidioidomycosis in hamsters (Mesocricetus auratus): Gestational interactions

Paracoccidioides brasiliensis is a dimorphic fungus presenting specific steroid hormone receptors, both in the yeast and mycelial forms and estrogen inhibits the transition from mycelium to yeast. In the acute phase, the disease occurs with equal frequency in both sexes but in adults, females are spared. Placental fungal infection has been reported, but references to fetal infection have not been confirmed. We used 78 Syrian female hamsters divided into 3 groups: GI consisted of 30 infected mated females, GII of 20 infected unmated females and GIII of 28 uninfected mated females. Animals of group I were mated 4 weeks after infection and half of them were submitted to cesarean section on day 15 after successful mating; the other half was maintained and submitted to cesarean section and sacrificed 14 weeks after infection. Half of the animals of group II were sacrificed seven weeks and the other half 14 weeks after infection. Uninfected animals of group III were treated the same as the animals of group I. The animals were infected with strain 18 of P. brasiliensis by the intracardiac route. We evaluated the disease by the volume of granulomas in different organs, number of fungi in liver and spleen and the immunologic responses [ELISA, Double Immunodifusion (DID), Delayed Hypersensitivity Skin Test (DHT) and Macrophage Migration Inhibition (MMI)]. We studied the infection through the gestation by evaluation of the abortions, morphologic and clinic examinations of the fetuses. Our results showed that the infection did not transfer to the fetus through the placenta, but the number of abortions was larger among infected females. The newborns of GI females were smaller, weighed less and showed little vitality. The disease was more severe and disseminated in infected mated females, especially in the second sacrifice 14 weeks after inoculation, when the total volume of granulomas in them (56.3 mm) was much greater than in the infected unmated females (12 mm).