Spermathecae of the mangrove crab Ucides cordatus: a histological and histochemical view

Ucides cordatus is the most commercially, important mangrove crab in Brazil. In spite of its economic importance, there are few studies of its reproduction, in particular the female reproductive system. The present study describes the histology and histochemistry of the spermathecae of U. cordatus. Adult females were can lit monthly from July 2004 through June 2005, at Iguape, State of São Paulo. The crabs were anaesthetized, and their spermathecae removed and fixed in Davidson's fluid, following the histological routine for paraffin. The slides were stained with HE, xylidine Ponceau, PAS, alcian blue (pH 1.0 and 2.5), Sudan black B and picrosirius-haematoxylin. Histologically, the spermathecae possesses a capsule of conjunctive tissue, rich in collalgen fibres, which surrounds the secretory columnar epithelium. In the lumen, individual sperm packets are not observed; the spermatophores are intermixed with the seminal fluid and secretions of the spermathecae itself. A large proportion of the free spermatozoids and spermatophores are arranged in homogeneous masses in the proximal part of the spermathecae. The secretion produced by the columnar epithelium appears to promote the movement of the gametes to the fertilization chamber, in a ventral position, allowing fertilization of the oocytes. Histochemically, the secretion produced by the columnar epithelium was strongly positive for neutral polysaccharides, positive for acid polysaccharides, and weakly positive for proteins and lipids. Tills secretion forms a glycoprotein matrix which is associated with maintenance of the spermatophores, which can remain stored for long periods.

Potential microbiological contamination of effluents in poultry and swine abattoirs

Health risks in the effluents of seven swine abattoirs and of seven poultry abattoirs were evaluated with regard to environment degradation and to dissemination of pathogenic microorganisms during the rainy and dry seasons. Supply-water samples from affluents and effluents of the treatment systems at different sites within the abattoir processing system were analysed. Similarly, water samples from the three recipient sites (emission point, 100 m upstream, 100 m downstream) were also analysed. Temperature, free residual chlorine (FRC), total coliform bacteria, Escherichia coli, enterococci, identification and serotyping of salmonellae were assessed. Scalding is the most significant stage in the slaughtering chain (P<0.05) when temperature is taken into account. Temperatures Lit effluents and at the sampled sites in the water bodies accorded to state and federal legislation standards. Supply waters did not meet the standards for FRC and microbial count standards according to the Ministry of Health and within limits imposed by the Industrial and Sanitary Inspection Regulations for Animal Products. Feather plucking and evisceration in Poultry slaughter and the cleansing of carcasses and facilities in Poultry and swine slaughtering had the highest contamination impact. The three loci at the water bodies were above the microbiological standards for classes II and III sites, in conformity with Law 8468 of the state of São Paulo, Brazil and Conama. Salmonella was found at several sites during slaughter, at both types of abattoirs, including in the effluent treatment system. This showed that these sites were the dissemination sources of the microorganism.

Severity of banana leaf spot in an intercropping system in two cycles of banana Prata Ana

Prata Ana is the most planted banana cultivar in northern Minas Gerais, Brazil. It is however susceptible to several pathogens. This study was carried out to evaluate the disease severity of banana leaf spot in the Prata Ana cv. in the first and second cycle under six different planting systems. The randomized block experimental design was used with six treatments and four replications. lit an evaluation of the severity of banana leaf spot, no disease symptoms were found on Thap Maeo and Caipira. The evolution curve of the disease indicated seasonal effects in the first and second cycles. The severity, of banana leaf spot was highest soon after the regional rainy period from November to March. A comparison of the means of the evaluations indicated a reduction in disease severity from the first to the second cycle.

Caracterização biológica, histopatológica e análise de ácido nucléico de uma cepa Trypanosoma cruzi da região de Marília, SP

Estudou-se o comportamento biológico e histopatológico de uma cepa genuínamente mariliense de Trypanosoma cruzi, isolada em 1997 através de xenodiagnóstico artificial. Vinte e cinco camundongos swiss foram infectados intraperitonealmente, sendo 11 utilizados para a realização da curva parasitêmica e observação da morfologia dos tripomastigotas e 14 foram sacrificados após o 17, 23, 30, 60 e 180 dias pós-infecção e coletados coração, esôfago, fígado, cólon, e músculo esquelético (fragmento da coxa direita) para análise histopatológica. Cultura em meio LIT foi realizada para análise de DNA. Os resultados mostraram predomínio de formas largas, baixa parasitemia com picos médios de 860 tripomastigotas/5mil de sangue ao redor do 20º dia de infecção. Nenhum camundongo morreu na fase aguda da infecção. Exame histopatológico mostrou poucos ninhos de amastigotas em coração, raros em músculo esquelético e cólon com discreto processo inflamatório. Comparada com a cepa Y, que foi isolada de uma paciente da mesma região, notamos diferentes características biológicas e comportamentais, porém a análise de DNA as coloca no mesmo grupo, demonstrando a proximidade dessas cepas.

Triatomíneos rupestres infectados por Trypanosomatidae, coletados em Quaraí, Rio Grande do Sul, 2003

Foram capturados triatomíneos rupestres em seis localidades do município de Quaraí-RS, com o intuito de verificar o índice de infecção por Trypanosomatidae, bem como o animal reservatório. A captura ocorreu no ambiente silvestre, sendo coletados 453 exemplares, os quais foram identificados e separados por estádio ninfal. Coletaram-se 421 (92,9%) exemplares de Triatoma rubrovaria, 26 (5,7%) de Triatoma circummaculata e 6 (1,3%) de Panstrongylus tupynambai. Dentre as três espécies coletadas, somente Triatoma rubrovaria mostrou-se positivo para Trypanosomatidae, num total de 13 (4,2%) exemplares. Após a inoculação em camundongos e meio de cultura LIT, isolaram-se cinco cepas de Trypanosoma cruzi. Dos triatomíneos infectados com Trypanosomatidae, 4 (30,8%) tiveram a reação de precipitina não reagente para os anti-soros testados, 4 (30,8%) positivos para anti-soro de roedor, 4 (30,8%) para anti-soro de cabra e 1 (7,7%) para anti-soro de porco e humano.

Definition of a diagnostic routine in individuals with inconclusive serology for chagas disease

Despite the existence of highly sensitive tests, inconclusive serological results are frequent in chronic chagasic infection. This study aimed to define a diagnostic conduct for 30 individuals with inconclusive serology (G3) for chagasic infection assisted at the Outpatient Unit for Infectious and Parasitic Diseases of the Botucatu School of Medicine. Twenty-one individuals with negative serology (G1) and 33 with positive serology (G2) were also studied. Serological methods ELISA, HAI, IFI and immunoblotting TESA-cruzi were used for G1, G2 and G3, and parasitological methods xenodiagnosis, hemoculture and PCR-LIT were used for G2 and G3 individuals. ELISA, HAI and IFI were performed in 5 different blood samples in G2 and G3. TESA-cruzi was carried out only once in G1, G2 and G3 and, since it is the most sensitive, it was utilized as standard. In G3, positivity for ELISA reached 86% in the fifth blood sample; the ELISA+HAI+IFI combination showed a maximum of 44.8% in the second sample; and TESA-cruzi, 76% in one single sample. Xenodiagnosis positivity was 9.4%; hemoculture showed 15.2%; and PCR-LIT exhibited 22% positivity in G2. Nevertheless, in G3, positivity percentage was 3.4% for xenodiagnosis, 6.7% for PCR-LIT, and no positive result was found for hemoculture. In G3, PCR-LIT resolved one case which was still inconclusive according to serology tests. In order to define inconclusive diagnoses, the results suggest the combined use of ELISA+HAI+IFI in 2 blood samples, decreasing the occurrence of false positive/negative results. If results remain inconclusive, the performance of TESA-cruzi and PCR-LIT, if necessary, is recommended.

The reproductive hiology and cycle of Rioraja agassizi (Muller and Henle, 1841) (Chondrichthyes : Rajidae) in southeastern Brazil, SW Atlantic Ocean

Specimens of Rioraja agassizi were collected Lit Guaruja pier, São Paulo State, Brazil, from March 2005 to March 2006. A total of: 275 males were captured. Based on the analysis of the clasper length, gonad weight, clasper gland length, alar thorns and lobule diameter, the size-at-maturity was estimated to be 32 cm. There were 1049 females in the record. Size-at-maturity was calculated in 40 cm. Sexual resting females were observed. Gonadosomatic and hepatosomatic indexes varied significantly in both sexes throughout the year. However, ovulation, egg-laying and presence of sperm in the seminal vesicle were observed all year round. For this reason, an annual cycle with at least one peak in the sexual activity is proposed for this species.

Tests to determine LC50 and discriminating doses for macrocyclic lactones against the cattle tick, Boophilus microplus

Laboratory tests were carried out on larvae and adults of the cattle tick Boophilus microplus to determine the toxicity of macrocyclic lactone acaricides (MLs). Technical and commercial MLs were used in larval packet test (LPT), larval immersion test (LIT) and adult immersion test (AIT). Tn LIT and AIT the toxicity of MLs was much higher than for LPT. In the AIT, diluting the injectable formulation of MLs in water was as effective as dilution in ethanol + Triton X-100. LC50, LC99.9 and 95% confidence limits were determined so that a discriminating dose (DD) could be set for larval and adult tests in order to diagnose potential resistance to MLs in field samples of the tick. These DDs are for Australian strains of B. microplus and may not be suitable for other strains until further work is carried out.The value of these diagnostic tests can only be verified if or when resistance to MLs emerges in ticks. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Locomotor and metabolic activities of Gymnostreptus olivaceus (Diplopoda, Spirostreptida) at different photoperiod conditions

In the present investigation the locomotor and the metabolic activity of Gymnostreptus olivaceus were studied, using 24-hr cycles at different photoperiods and constant temperature and RH. Locomotor activity was studied by the actography method and was reported as coefficients of nocturnalism [CN (N/N + D). 100]. The results showed a nocturnalism coefficient of 98,71% under normal photoperiod conditions and of 29,58% under inverted photoperiod conditions. In constant illumination, the CN of G. olivaceus was 88,22%, and in constant darkness, its rhythm was close to that of the normal photoperiod (CN = 94,92%). The metabolic activity was studied by manometric Warburg respirometry and lit was reported as mu l O-2 . g(-1). h(-1). The respiratory rate of G. olivaceus under normal photoperiod condition was higher at night than during the day (52,52 x 28,76), coinciding with the pattern of nocturnal locomotor activity of the animal. However, under conditions of inverted photoperiod, the millipede maintained its tendency toward a more intense nocturnal respiratory rate (50,35 x 39,14). Similar behaviours were observed under constant illumination and constant darkness, in which G. olivaceus again presented higher nocturnal respiratory rates than diurnal ones(85,84 x 53,48 and 73,18 x 57,0, respectively). The present experimental data suggests the persistence of an endogenous rhythm where the light may not be an important exogenous synchronizer of the activity of G. olivaceus, because it was insufficient to block the start of the biological clock and the natural tendency of higher nocturnal activities of millipedes, principally when the tests were performed in constant illumination or darkness (free-running tests).

Crystallization of piratoxin I, a myotoxic LYS49-phospholipase A(2) homologue isolated from the venom of Bothrops pirajai

Fernanda Canduri, Lit C. Mancuso, Andreimar M. Soares, Jose R. Giglio, Richard J. Ward and Raghuvir K. Arni. Crystallization of piratoxin I, a myotoxic Lys49-phospholipase A(2) homologue isolated from the venom of Bothrops pirajai. Toxicon 36, 547-551, 1998.-Large single crystals of piratoxin I, a Lys49-PLA(2) homologue with low enzymatic activity, have been obtained. The crystals belong to the orthorhombic system space group p2(1)2(1)2(1) and diffract X-raps to a resolution of 2.8 Angstrom. Preliminary analysis reveals the presence of two molecules in the crystallographic asymmetric unit. (C) 1998 Elsevier B.V. Ltd. All rights reserved.

Histochemical and ultrastructural cytochemistry of glycogen on ovarioles of Neoponera villosa ants (Hymenoptera: Ponerinae)

In Neoponera villosa ants, we found ovaries of the polytrophic meroistic type which is characterized by the presence of nurse cells forming together with the oocyte, the so-called follicles. The nurse cells have the primary function of supplying the oocyte with RNA, but they contribute to the supply of other elements such as glycogen. With the objetive of detecting the presence of this substance in the ovarioles of workers and queens of N.villosa ante the ovaries were removed and processed according to electron microscopy technic for glycogen detection. Glycogen is a common element in insect oocytes and is abundantly distributed in the cytoplasm of N.villosa workers and queens. However, in ovarian follicles it can only be detected at stages ET and lit of development. Glycogen synthesis probably occurs predominantly in nurse cells which transfer it into the oocyte through the nourish pore. This process requires high energy expenditure that justify the large numbers of mitochondria associated with glycogen in the nurse cell cytoplasm. The amount of glycogen in the nurse cells of queens is slightly greater than workers.

Avaliação da técnica de hemocultura pela reação em cadeia da polimerase (PCR) utilizando os iniciadores P35/P36 e TCZ1/TCZ2 para a detecção de Trypanosoma cruzi em cães e gatos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Hemocultura, reação de imunofluorescência indireta (RIFI) e reação em cadeia pela polimerase (PCR) para Leishmania spp. em cães e gatos provenientes de área endêmica e não endêmica para leishmaniose

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