Description of eggs and nymphal instars of Triatoma baratai Carcavallo & Jurberg, 2000 based on optical and scanning electron microscopy (Hemiptera: Reduviidae: Triatominae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Morphology of the eggs of the genus Belminus (Hemiptera: Reduviidae: Triatominae) by optical and scanning electron microscopy

The external morphology of eggs of the species Belminus corredori Galvao & Angulo, 2006, Belminus herreri Lent & Wygodzinsky, 1979 and Belminus ferroae Sandoval, Pabon, Jurberg & Galvao, 2007 is described for the first time using scanning electron microscopy (SEM). Females lay one egg at a time; the shape of the egg is oval with lateral flattening, a rounded caudal area and a true and simple convex operculum. The average size of the egg among these species is 0.51-0.58 mm in length and 0.36-0.4 mm in width. As a generic character we highlight the discovery of a chorion rim with one to four grooves always joined to the micropyles. All species present a clear polygonal pattern only in the operculum and in the egg's cephalic region, while the median and caudal regions show a smooth appearance, features that are shared only with the eggs of the genus Alberprosenia. A discriminant analysis of the polygonal design of the operculum proved to be useful for the identification of the species.

Improved laboratory safety by decontamination of unstained sputum smears for acid-fast microscopy

Tubercle bacilli may survive in unstained heat-fixed sputum smears and may be an infection risk to laboratory staff. We compared the effectiveness of 1% and 5% sodium hypochlorite, 5% phenol, 2% glutaraldehyde, and 3.7% formalin in killing Mycobacterium tuberculosis present in smears prepared from 51 sputum samples. The smears were decontaminated by the tube and slide techniques. Phenol at 5%, glutaraldehyde at 2%, and buffered formalin at 3.7% for 1 min (tube technique) or for 10 min (slide technique) were effective in decontaminating sputum smears and preserved cell morphology and quantitative acid-fast microscopy results.

Direct observation of oxidative stress on the cell wall of Saccharomyces cerevisiae strains with atomic force microscopy

We imaged pores on the surface of the cell wall of three different industrial strains of Saccharomyces cerevisiae using atomic force microscopy. The pores could be enlarged using 10 mM diamide, an SH residue oxidant that attacks surface proteins. We found that two strains showed signs of oxidative damage via changes in density and diameter of the surface pores. We found that the German strain was resistant to diamide induced oxidative damage, even when the concentration of the oxidant was increased to 50 mM. The normal pore size found on the cell walls of American strains had diameters of about 200nm. Under conditions of oxidative stress the diameters changed to 400nm.This method may prove to be a useful rapid screening process (45-60 min) to determine which strains are oxidative resistant, as well as being able to screen for groups of yeast that are sensitive to oxidative stress. This rapid screening tool may have direct applications in molecular biology (transference of the genes to inside of living cells) and biotechnology (biotransformations reactions to produce chiral synthons in organic chemistry.

Light microscopy and ultrastructure of the liver of Astyanax altiparanae Garutti and Britski, 2000 (Teleostei, Characidae).

Livers of thirty specimens of Astyanax altiparanae obtained from a commercial fish farm were subjected to light and transmission electron microscopy, in order to describe the hepatic parenchyma and the intrahepatic exocrine pancreatic tissue. Anatomically, the liver showed only three hepatic lobes. Histological analysis demonstrated that the hepatocytes were spread out as anastomotic cords, arranged in two cellular layers and surrounded by sinusoids. The intrahepatic exocrine pancreatic tissue exhibited an acinar arrangement and was diffused in the hepatic parenchyma. Ultrastructural analysis showed that the hepatocytes had a rounded nucleus and a rough endoplasmatic reticulum, with a parallel disposition to the nuclear membrane. The exocrine pancreatic cells showed secretion granules at the apical portion, and the rough endosplasmatic reticulum was concentrically distributed.

Quantitative fractography under light microscopy: A digital image processing approach

This work is an example of the improvement on quantitative fractography by means of digital image processing and light microscopy. Two techniques are presented to investigate the quantitative fracture behavior of Ti-4Al-4V heat-treated alloy specimens, under Charpy impact testing. The first technique is the Minkowski method for fractal dimension measurement from surface profiles, revealing the multifractal character of Ti-4Al-4V fracture. It was not observed a clear positive correlation of fractal values against Charpy energies for Ti-4Al-4V alloy specimens, due to their ductility, microstructural heterogeneities and the dynamic loading characteristics at region near the V-notch. The second technique provides an entire elevation map of fracture surface by extracting in-focus regions for each picture from a stack of images acquired at successive focus positions, then computing the surface roughness. Extended-focus reconstruction has been used to explain the behavior along fracture surface. Since these techniques are based on light microscopy, their inherent low cost is very interesting for failure investigations.

Digital Light Microscopy of Macroporous Ceramics Processed by Consolidation Casting with Mixtures of Native Starches

Porosity in starch consolidation casting technique is rightly related to original size and morphology of starch granules, leaving a pore structure after burning out. This work reports the results for the addition of different native potato and corn starch proportions in suspension,; with TiO(2) (rutile) powder. Gelling temperature have been defined after observation under light microscopy using a heating stage. Analysis of porous network and isolated pores have been clone from images of samples surfaces obtained by depth from focus reconstruction, revealing a qualitative correlation of pores characteristics and starches additions in suspensions, suggesting that the presence of isolated or interconnected pores can be handled by starches selection to control the amylopectin and amylose contents in slurries. Also, the analysis of porous fraction distribution shows no consistent pattern through specimens' volume according to starches in mixtures.

Low voltage and variable-pressure scanning electron microscopy of fractured composites

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

Quantitative microscopy characterization of hydrous niobium phosphate into bleached cellulose

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

Diagnosis of clinical bovine mastitis by fine needle aspiration followed by staining and scanning electron microscopy in a Prototheca zopfii outbreak

Biopsy by fine needle aspiration together with microbiological examination and scanning electron microscopy were evaluated in diagnosis of clinical bovine mastitis in a Prototheca zopfii outbreak. Fine needle aspiration was performed in 21 mammary quarters from ten Holstein cows presenting clinical mastitis caused by P. zopfii. The algae were previously identified in the microbiological examination of milk collected from these cows. Material aspirated from these 21 mammary glands was submitted to cytological staining (Gram, Giemsa and/or Shor staining). Fine needle aspiration enabled cytological identification of the algae in these 21 mammary glands, from which P. zopfii was isolated in the milk. Simultaneously, five mammary fragments collected by fine needle aspiration from these 21 mammary glands presenting clinical mastitis were also submitted to microbiological examination. P. zopfii was also isolated from these five fragments. Scanning electron microscopy technique also identified three of these five P. zopfii strains isolated from mammary fragments collected by cytological aspiration. These results suggest that fine needle aspiration may be an alternative method for the diagnosis of clinical mastitis.

Surveillance of canine visceral leishmaniasis in a disease-free area

A leishmaniose é uma importante parasitose re-emergente observada no mundo, particularmente em países tropicais. Não há ainda relatos de casos autóctones no estado do Paraná. Não há até o momento referência de vigilância no reservatório canino, tais como Curitiba e região metropolitana do estado. O objetivo do estudo foi determinar a soroprevalência da leishmaniose visceral em cães entregues ao Centro de Controle de Zoonoses de São José dos Pinhais, Paraná para eutanásia. A detecção sorológica da presença de anticorpos contra Leishmania sp. foi realizada por (ELISA) indireto e pela Reação de Imunofluorescência Indireta (RIFI). Além disso, impressão de linfonodo poplíteo coletadas ao acaso de 50 cães com sinais clínicos suspeitos para leishmaniose visceral e analisados sob microscopia óptica para detecção de formas amastigotas, foram negativas. Amostras de soro de 364 animais foram testadas, e os resultados mostraram somente uma amostra positiva (0,0027%), reagente ao ELISA e negativa à RIFI, entretanto, o cão não apresentava sinais clínicos. A vigilância ao acaso em uma população de vários locais de uma área metropolitana pode ser uma forma de prevenção da disseminação da doença. Com base nos resultados observados, Curitiba e região metropolitana foram consideradas de baixo risco para a leishmaniose visceral.

The use of transmission electron microscopy and oocyte transfer to evaluate in vitro maturation of equine oocytes in different culture conditions

The current study evaluates the ability of equine oocytes matured in different conditions to undergo nuclear and cytoplasmic maturation.. After oocyte transfer, embryonic development was diagnosed at 1.5 and 90 days of gestation. For each group, immature oocytes obtained from slaughterhouse ovaries were matured in vitro (5 replicates). In experiment I, three different media were tested. HTF:BME, SOFaa, and TCM 199. In experiment 11, the HTF:BME was chosen as maturation medium containing pFSH, eFSH, or eFSH + eGH. Nuclear maturation was estimated after stripping the oocytes and staining with Hoechst 33342. The evaluation of cytoplasmic maturation was performed by transmission electron microscopy. For oocyte transfer, six non-cycling recipient mares were used, and 8 to 15 oocytes were transferred in each mare. In experiment I, the results showed no differences (P > .05) in nuclear maturation (MII) among experimental groups. The percentage of MII was 29.3 ( +/- 9.6), 23.4 ( +/- 8.4), and 13.5 ( +/- 12.4) for HTF:BME, SOF, and TCM, respectively. In experiment II, all media tested were efficient in inducing metaphase II. Also, no statistical differences (P > .05) were observed in percentages of nuclear maturation rates when porcine (37.1 +/- 22.4) or equine (25.8 +/- 8.2) FSH were used, or when eFSH + eGH was added to HTF:BME (29.4 +/- 12.3). The analysis of cytoplasmic morphology of oocytes cultured in TCM 199 and SOFaa showed signs of incomplete cytoplasmic maturation and premature cortical reaction. Meanwhile, oocytes cultured in HTF:BME medium presented cytoplasmic characteristics similar to those described by others for in vivo-matured oocytes. The addition of eFSH to the HTF:BME medium resulted in an improvement of cytoplasmic morphology. After oocyte transfer, two mares became pregnant, one from pFSH group and one from eFSH+eGH group. These results indicate that although in vitro matured equine oocytes are capable of fertilization and embryonic development, the percentage of competent oocytes is still low.