Spontaneous poisoning by larvae of Perreyia flavipes (Pergidae) in sheep

From a flock of 175 Texel sheep 25 animals died after consumption of a sawfly larvae subsequently identified as Perreyia flavipes. The disease occurred in June-July 2006 on a farm located in the county of Encruzilhada do Sul, Rio Grande do Sul, Brazil. Although there were 11 cattle in the same paddock, none of them was affected. High numbers of compact masses containing up to 150 larvae were scattered in the paddock where the animals were grazing. Most affected sheep showed severe apathy during 24-36 h before death, but weakness, muscular tremors and depression were also observed. Necropsy was performed on six sheep and the main macroscopic lesions were hemorrhages in the subcutaneous tissues, endocardium, gallbladder wall, and abomasal mucosa. In all animals was found hydrothorax, hydropericardium, ascites, and mild jaundice. Edema in the abomasal folds, mesentery, perirenal tissues, and gallbladder wall were also seen. The livers were yellowish with disseminated pinpoint hemorrhages in the parenchyma and had an enhanced lobular pattern. Perreyia flavipes larval body fragments and heads were found in the forestomach contents of the six sheep. Feces were scant, dry and formed balls coated by mucus and streaks of blood. Similar contents were also present at the end of the cecum. Prominent microscopic lesions included severe and diffuse periacinar or massive necrosis of hepatocytes associated with multifocal random hemorrhages. Diffuse necrosis of lymphoid follicles in lymph nodes and Peyer's patches, lymphoid depletion and necrosis in germinative centers of the spleen, and diffuse vacuolization in the renal tubular epithelia were also seen.

Gastric ulcers in pigs affected with postweaning multisystemic wasting syndrome

Samples of gastric lymph nodes and the stomachs from 24 pigs selected from herds affected by postweaning multisystemic wasting syndrome (PMWS) and sudden death associated with gastric ulcers were studied. Pigs were selected on the basis of unthriftiness, decreased feed intake, and wasting. The stomachs were opened, inverted, and classified into 0-3 score according the severity of the gross lesions present in pars oesophagica (non-glandulargastric mucosa). Selected samples were processed for paraffin embedding and stained with hematoxylin and eosin. Immunohistochemistry using anti-PCV2 (porcine circovírus type 2) antibody, anti-Helicobacter pylori antibody and a wide-spectrum anti-cytokeratin antibody was performed. Gross changes in pars oesophagea were classified according to the severity of lesions as score 3, 2, and 1 in 8, 6, 5 stomachs respectivelly. Microscopically, hyperplastic lymphoid follicles, lymphohistiocytic inflammatory infiltrates and focci of necrosis in the gastric mucosa were common findings. Large amounts of PCV2 antigen were observed in the cytoplasm and nuclei from intralesional cells and debris from the gastric glandular mucosal zone; however, in the fundus, anti-PCV2 immunostaining was restricted to the surface mucosal cells and foveolar compartment. All gastric lymph nodes were positive for PCV2 antigen. Anti-H. pylori immunostaining was seen in eleven cases, mainly in the antrum, on the mucosal surface and foveolar compartment. The association of the anti-PCV2 immunostaining with the glandular mucus-producing cells suggests a role for PCV2 as an additional factor for the swine ulcer development.

Pressure drop coefficients for elliptic and circular sections in one, two and three-row arrangements of plate fin and tube heat exchangers

The objective of the present work is the experimental determination of pressure drop coefficients (loss coefficients) for elliptic and circular sections in one, two and three-row arrangements of plate fin and tube heat exchangers. The experiments permitted to correlate the dimensionless loss coefficient with the flow Reynolds number in the rectangular channel formed by the plate fins. The experimental technique consisted of the measurement of the longitudinal pressure distribution along the flow channel, for several values of air mass flow rate. The total number of data runs, each one characterized by the flow Reynolds number, was 216. The present geometry is used in compact heat exchangers for air conditioning systems, heaters, radiators, and others. Also, it is verified the influence of the utilization of elliptic tubes, instead of circular ones, in the pressure drop. The measurements were performed for Reynolds numbers ranging from 200 to 1900.

Dynamics of coherent vortices in mixing layers using direct numerical and large-eddy simulations

Coherent vortices in turbulent mixing layers are investigated by means of Direct Numerical Simulation (DNS) and Large-Eddy Simulation (LES). Subgrid-scale models defined in spectral and physical spaces are reviewed. The new "spectral-dynamic viscosity model", that allows to account for non-developed turbulence in the subgrid-scales, is discussed. Pseudo-spectral methods, combined with sixth-order compact finite differences schemes (when periodic boundary conditions cannot be established), are used to solve the Navier- Stokes equations. Simulations in temporal and spatial mixing layers show two types of pairing of primary Kelvin-Helmholtz (KH) vortices depending on initial conditions (or upstream conditions): quasi-2D and helical pairings. In both cases, secondary streamwise vortices are stretched in between the KH vortices at an angle of 45° with the horizontal plane. These streamwise vortices are not only identified in the early transitional stage of the mixing layer but also in self-similar turbulence conditions. The Re dependence of the "diameter" of these vortices is analyzed. Results obtained in spatial growing mixing layers show some evidences of pairing of secondary vortices; after a pairing of the primary Kelvin-Helmholtz (KH) vortices, the streamwise vortices are less numerous and their diameter has increased than before the pairing of KH vortices.

Weed suppression in the formation of brachiarias under three sowing methods

The success of conservation systems such as no-till depends on adequate soil cover throughout the year, which is possible through the use of cover crops. For this purpose the species belonging to the genus Urochloa has stood out by virtue of its hardiness and tolerance to drought. Aiming ground cover for the no-till system, the objective was to evaluate the establishment of two species of the genus Urochloa, in three sowing methods, in the weed suppression and the sensitivity of these forages to glyphosate. The study design was a randomized block with a 2 x 3 x 3 factorial arrangement, in which factor A was composed of Urochloa ruziziensis and Urochloa hybrid CIAT 36087 cv. Mulato II, factor B was formed by sowing methods: sown without embedding, sown with light embedding and sown in rows, and factor C was composed of three doses of glyphosate (0.975, 1.625 and 2.275 kg ha-1 of acid equivalent). For determination of weed suppression, assessment of biomass yield and soil cover was performed, by brachiaria and weeds, at 30, 60, 90, 120 and 258 days after sowing. Visual assessment of the desiccation efficiency at 7 and 14 days after herbicide application was performed. It is concluded that embedding Urochloa seeds stands out in relation to sowing in the soil surface. Urochloa ruziziensis is more efficient in the dry weight yield, weed suppression, in addition to being more sensitive to glyphosate herbicide.

Sunflower response to repeated foliar applications of Paclobutrazol

Paclobutrazol (PBZ), a well-known growth retardant of the triazole family, is mostly used for controlling plant size and growth, resulting in more desirable compact plants for floricultural purposes; however, limited experimental data are available for use of PBZ in field sunflower. The objective of this study was to evaluate the effect of repeated foliar applications of PBZ at rates of 50 + 50 g ha-1(double application) and 50 + 50 + 50 g ha-1 (triple application) on sunflower morphology and productivity. PBZ applications corresponded to growth stages of four to five, five to six, and six to eight true leaves of sunflower plants, respectively. The double foliar application of PBZ reduced sunflower plant height at maturity by 4.4% (or by 11.7 cm). The triple foliar application of PBZ reduced sunflower plant height at maturity by 14.4% (or by 49.2 cm). However, PBZ either in the double or the triple application reduced achene yield per plant by 25.6% and 22.5% and the 100-achene weight by 11.4% and 25.0%, respectively, compared with the non-treated control. Overall, the repeated foliar applications of PBZ at the rates tested in this study, apart from providing a reduction of sunflower plant height, had major adverse effects on achene yield and weight. Thus, different application schemes of PBZ or perhaps different growth regulators should be investigated to control plant height in sunflower.

Vegetative propagation strategies of four rupestrian species of Leiothrix (Eriocaulaceae)

Leiothrix is endemic of South America and includes 37 species, 25 of which occur in the state of Minas Gerais. Nineteen of those occur in the "Serra do Cipó", a mountain chain, located in the southern portion of the Espinhaço mountain range. This study examines vegetative propagation strategies of four species of Leiothrix, endemic to the Minas Gerais portion of the Espinhaço mountain range. For each species we established permanent plots, where we marked 30 to 51 rosettes or clones, and then took morphological and phenological measurements. Leiothrix crassifolia (Bong.) Ruhland and L. curvifolia var. lanuginosa (Bong.) Ruhland are rhizomatous, forming compact clones. Leiothrix vivipara (Bong.) Ruhland does not produce rhizomes, but is pseudoviviparous, i.e., produces numerous ramets originating from inflorescences. These ramets are formed precociously, and the flower heads do not touch the ground. In Leiothrix spiralis (Bong.) Ruhland both of these strategies are seen: it is both rhizomatous and pseudoviviparous. In this species, the ramets are formed late, only after the flower head has touched the ground. One of the typical conditions of the rupestrian grasslands is soil water shortage in some periods of the year and nutrient scarcity all year round. These conditions might have created an ideal ecological scenario for the evolution of both pseudovivipary and rhizomatous clonal growth in Leiothrix.

Techniques of radioautography for medical and biological research

Standard techniques for radioautography used in biological and medical research can be classified into three categories, i.e., macroscopic radioautography, light microscopic radioautography and electron microscopic radioautography. The routine techniques used in these three procedures are described. With regard to macroscopic radioautography, whole body radioautography is a standard technique which employs freezing and cryosectioning and can demonstrate organ distributions of both soluble and insoluble compounds. In contrast, in light and electron microscopic radioautography, soluble and insoluble techniques are separated. In order to demonstrate insoluble labeled compounds, conventional chemical fixations such as formalin for light microscopy or buffered glutaraldehyde and osmium tetroxide for both light and electron microscopy followed by dehydration, embedding and wet-mounting applications of radioautographic emulsions can be used. For the demonstration of soluble labeled compounds, however, cryotechniques such as cryofixation, cryosectioning, freeze-drying, freeze-substitution followed by dry-sectioning and dry-mounting radioautography should be employed both for light and electron microscopy. The outlines of these techniques, which should be utilized in various fields of biological and medical research, are described in detail

Cytokine accumulation in osteitis fibrosa of renal osteodystrophy

Bone marrow fibrosis occurs in association with a number of pathological states. Despite the extensive fibrosis that sometimes characterizes renal osteodystrophy, little is known about the factors that contribute to marrow accumulation of fibrous tissue. Because circulating cytokines are elevated in uremia, possibly in response to elevated parathyroid hormone levels, we have examined bone biopsies from 21 patients with end-stage renal disease and secondary hyperparathyroidism. Bone sections were stained with antibodies to human interleukin-1alpha (IL-1alpha), IL-6, IL-11, tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-ß (TGF-ß) using an undecalcified plastic embedding method. Intense staining for IL-1alpha, IL-6, TNF-alpha and TGF-ß was evident within the fibrotic tissue of the bone marrow while minimal IL-11 was detected. The extent of cytokine deposition corresponded to the severity of fibrosis, suggesting their possible involvement in the local regulation of the fibrotic response. Because immunoreactive TGF-ß and IL-6 were also detected in osteoblasts and osteocytes, we conclude that selective cytokine accumulation may have a role in modulating bone and marrow cell function in parathyroid-mediated uremic bone disease.

DNA-lipid systems: A physical chemistry study

It is well known that the interaction of polyelectrolytes with oppositely charged surfactants leads to an associative phase separation; however, the phase behavior of DNA and oppositely charged surfactants is more strongly associative than observed in other systems. A precipitate is formed with very low amounts of surfactant and DNA. DNA compaction is a general phenomenon in the presence of multivalent ions and positively charged surfaces; because of the high charge density there are strong attractive ion correlation effects. Techniques like phase diagram determinations, fluorescence microscopy, and ellipsometry were used to study these systems. The interaction between DNA and catanionic mixtures (i.e., mixtures of cationic and anionic surfactants) was also investigated. We observed that DNA compacts and adsorbs onto the surface of positively charged vesicles, and that the addition of an anionic surfactant can release DNA back into solution from a compact globular complex between DNA and the cationic surfactant. Finally, DNA interactions with polycations, chitosans with different chain lengths, were studied by fluorescence microscopy, in vivo transfection assays and cryogenic transmission electron microscopy. The general conclusion is that a chitosan effective in promoting compaction is also efficient in transfection.

DNA extraction and quantification from touch and scrape preparations obtained from autopsy liver cells

The objective of the present study was to develop a simplified low cost method for the collection and fixation of pediatric autopsy cells and to determine the quantitative and qualitative adequacy of extracted DNA. Touch and scrape preparations of pediatric liver cells were obtained from 15 cadavers at autopsy and fixed in 95% ethanol or 3:1 methanol:acetic acid. Material prepared by each fixation procedure was submitted to DNA extraction with the Wizard® genomic DNA purification kit for DNA quantification and five of the preparations were amplified by multiplex PCR (azoospermia factor genes). The amount of DNA extracted varied from 20 to 8,640 µg, with significant differences between fixation methods. Scrape preparation fixed in 95% ethanol provided larger amount of extracted DNA. However, the mean for all groups was higher than the quantity needed for PCR (50 ng) or Southern blot (500 ng). There were no qualitative differences among the different material and fixatives. The same results were also obtained for glass slides stored at room temperature for 6, 12, 18 and 24 months. We conclude that touch and scrape preparations fixed in 95% ethanol are a good source of DNA and present fewer limitations than cell culture, tissue paraffin embedding or freezing that require sterile material, culture medium, laboratory equipment and trained technicians. In addition, they are more practical and less labor intensive and can be obtained and stored for a long time at low cost.

5-Bromo-2’-deoxyuridine induces visible morphological alteration in the DNA puffs of the anterior salivary gland region of Bradysia hygida (Diptera, Sciaridae)

5-Bromo-2’-deoxyuridine (BrdUrd) has long been known to interfere with cell differentiation. We found that treatment ofBradysia hygida larvae with BrdUrd during DNA puff anlage formation in the polytene chromosomes of the salivary gland S1 region noticeably affects anlage morphology. However, it does not affect subsequent metamorphosis to the adult stage. The chromatin of the chromosomal sites that would normally form DNA puffs remains very compact and DNA puff expansion does not occur with administration of 4 to 8 mM BrdUrd. Injection of BrdUrd at different ages provoked a gradient of compaction of the DNA puff chromatin, leading to the formation of very small to almost normal puffs. By immunodetection, we show that the analogue is preferentially incorporated into the DNA puff anlages. When BrdUrd is injected in a mixture with thymidine, it is not incorporated into the DNA, and normal DNA puffs form. Therefore, incorporation of this analogue into the amplified DNA seems to be the cause of this extreme compaction. Autoradiographic experiments and silver grains counting showed that this treatment decreases the efficiency of RNA synthesis at DNA puff anlages.

Morphometric analysis of the phrenic nerve in male and female Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR)

Ventilatory differences between rat strains and genders have been described but the morphology of the phrenic nerve has not been investigated in spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) rats. A descriptive and morphometric study of the phrenic nerves of male (N = 8) and female (N = 9) SHR, and male (N = 5) and female (N = 6) WKY is presented. After arterial pressure and heart rate recordings, the phrenic nerves of 20-week-old animals were prepared for epoxy resin embedding and light microscopy. Morphometric analysis performed with the aid of computer software that took into consideration the fascicle area and diameter, as well as myelinated fiber profile and Schwann cell nucleus number per area. Phrenic nerves were generally larger in males than in females on both strains but larger in WKY compared to SHR for both genders. Myelinated fiber numbers (male SHR = 228 ± 13; female SHR = 258 ± 4; male WKY = 382 ± 23; female WKY = 442 ± 11 for proximal right segments) and density (N/mm²; male SHR = 7048 ± 537; female SHR = 10355 ± 359; male WKY = 9457 ± 1437; female WKY = 14351 ± 1448) for proximal right segments) were significantly larger in females of both groups and remarkably larger in WKY than SHR for both genders. Strain and gender differences in phrenic nerve myelinated fiber number are described for the first time in this experimental model of hypertension, indicating the need for thorough functional studies of this nerve in male and female SHR.

Effect of different proportions of brea gum in the functional characteristics of wheat flour starch: impact on the physical quality of bread

The aim of this work was to study the changes induced by BG in the behaviour of wheat starch, and observe the influence of these variations on the quality of a basic white bread. The effect of four BG addition levels in the wheat flour functional characteristics (WAI, WSI, and pasting properties) and bread quality (physical parameters, crumb grain structure, moisture and hardness) was investigated. The highest levels of BG (1% and 2%) decreased the peak viscosity, and increased the stability and setback of the flour. This was due to a lower gelatinization of the starch granules, caused by a competition for water between the hydrocolloid and starch. These changes influenced the bread quality. The loaves added with 1% and 2% of BG presented smaller alveoli: this resulted in more compact, hard and less airy crumbs. Nevertheless, the moisture of the samples at 1% and 2% of added gum was higher than the control bread. However, the incorporation of BG at 0.5% did not affect the pasting parameters and bread quality, but increased moisture of crumb, so this concentration would be most recommended for baking, since higher humidity could favour the shelf- life of the product.