Análise espermática de perdizes (Rhynchotus rufescens) criadas em cativeiro e suplementadas com selênio

Due to the commercial importance of the red-winged tinamou (Rhynchotus rufescens), for the past few years, the employment of reproductive biotechnologies has been attempted. Thirty animals were randomly assigned into two groups: control group (no selenium) and treatment group (supplemented with 0,2 a 0,8 mg selenium/ 1000 kg ration). Animals were allocated at the FCAV - UNESP/Jaboticabal (2007-2008). Semen collections were performed by digital manipulation and divided in pools of at least 150 μL. After the immediate evaluation of motility, vigour, concentration and morphology, an aliquot of 20 μL was diluted in 300 μL of physiologic solution in order to test acrosome and membrane integrities, which were performed by counting 200 cells for each test. Cells were evaluated as follows: 1) Intact acrosome: lilac acrosome; Non-intact acrosome: pink acrosome; 2) Live cells: non stained; Dead: stained. Data was statistically analysed using the SAS System for Windows. No differences were found between treatment and control groups for volume, motility, vigour, mean number of spermatozoa per animal, concentration, Intact acrosome, Intact membrane. The difference found on midpiece sperm defect (Se = 1,33 ± 0,53 and control = 3,78 ± 0,69, p = 0.0107) may be due to the damages caused by the selenium deficiency to the architecture of the midpiece, which compromises sperm mobility and fertilization capacity.

Influence of dental exposure to oral environment on smear layer removal and collagen exhibition after using different conditioning agents

Although in vitro studies have shown encouraging results for root surface conditioning with demineralizing agents, in vivo studies have failed to show its benefits in periodontal healing. This can be attributed to several factors, among which, the hypermineralization of dental surface. Therefore, this in vitro study compared, using scanning electron microscopy (SEM), the effect of root surface conditioning with different conditioners (1% and 25% citric acid, 24% EDTA and 50 mg/mL tetracycline hydrochloride) in impacted teeth and in teeth that had their roots exposed to the oral environment. One trained examiner assessed the SEM micrographs using a root surface modification index. There was a tendency of more root surface modification in the group of impacted teeth, suggesting that the degree of root mineralization influences its chemical demineralization.

In vitro effect of low-level laser therapy on typical oral microbial biofilms

The aim of this study was to evaluate the effect of specific parameters of low-level laser therapy (LLLT) on biofilms formed by Streptococcus mutans, Candida albicans or an association of both species. Single and dual-species biofilms - SSB and DSB - were exposed to laser doses of 5, 10 or 20 J/cm 2 from a near infrared InGaAsP diode laser prototype (LASERTable; 780 ± 3 nm, 0.04 W). After irradiation, the analysis of biobilm viability (MTT assay), biofilm growth (cfu/mL) and cell morphology (SEM) showed that LLLT reduced cell viability as well as the growth of biofilms. The response of S. mutans (SSB) to irradiation was similar for all laser doses and the biofilm growth was dose dependent. However, when associated with C. albicans (DSB), S. mutans was resistant to LLLT. For C. albicans, the association with S. mutans (DSB) caused a significant decrease in biofilm growth in a dose-dependent fashion. The morphology of the microorganisms in the SSB was not altered by LLLT, while the association of microbial species (DSB) promoted a reduction in the formation of C. albicans hyphae. LLLT had an inhibitory effect on the microorganisms, and this capacity can be altered according to the interactions between different microbial species.

Homologies of the flower and inflorescence in the early-divergent grass anomochloa (poaceae)

Premise of the study: The grass subfamily Anomochlooideae is phylogenetically significant as the sister group to all other grasses. Thus, comparison of their structure with that of other grasses could provide clues to the evolutionary origin of these characters. Methods: We describe the structure, embryology, and development of the flower and partial inflorescence of the monotypic Brazilian grass Anomochloa marantoidea. We compare these features with those of other early-divergent grasses such as Pharus and Streptochaeta and closely related Poales such as Ecdeiocolea. Key results: Anomochloa possesses several features that are characteristic of Poaceae, notably a scutellum, a solid style, reduced stamen number, and an ovary with a single ovule that develops into a single indehiscent fruit. Interpretation of floral patterning in Anomochloa is problematic because the ramification pattern of the florets places the bracts and axes in unusual positions relative to the primary inflorescence axis. Our study indicates that there is a single abaxial carpel in Anomochloa, probably due to a cryptic type of pseudomonomery in Anomochloa that resembles the pseudomonomery of other grasses. On the other hand, the Anomochloa flower differs from the typical grass flower in lacking lodicules and possessing four stamens, in contrast with the tristaminate condition that characterizes many other grasses. Conclusions: Using the median part of the innermost bract as a locator, we tentatively homologize the inner bract of the Anomochloa partial inflorescence with the palea of other grasses. In this interpretation, the pattern of monosymmetry due to stamen suppression differs from that of Ecdeiocolea. © 2012 Botanical Society of America.

Blood cells attachment after root conditioning and PRP application: An in vitro study

Aim: Root conditioning is aimed at smear layer removal and at dental matrix collagen exposure, which may promote periodontal regeneration. This in vitro study assessed smear layer removal, collagen fiber exposure and the influence of PRP (platelet-rich plasma) application on adhesion of blood cells to the root surface using scanning electron microscopy (SEM). Materials and methods: Scaled root samples (n = 160) were set in five groups and conditioned with: group I - control group (saline solution); group II (EDTA 24%); group III (citric acid 25%); group IV (tetracycline hydrochloride 50 mg/ml); group V (sodium citrate 30%). Eighty samples were assessed using the root surface modification index (RSMI). The other eighty samples were set in two groups. The first group (n = 40) received PRP gel application with a soft brush and the second group (n = 40) received PRP application and then a blood drop. The fibrin clot formation was assessed in the first group and the blood cells adhesion was assessed in the second group using the BEAI (blood elements adhesion index). A previously trained, calibrated, and blind examiner evaluated photomicrographs. Statistical analysis was performed using the Kruskal-Wallis's and Dunn's tests. Results: Group III attained the best results for RSMI and BEAI. Moreover, it was the only group showing fibrin clot formation. Conclusion: Citric acid was the most efficient conditioner for smear layer removal, collagen fiber exposure and blood cell adhesion. Moreover, it was the only group showing fibrin clot formation after PRP application. Clinical significance: This study demonstrated that root conditioning followed by PRP application may favor blood cell adhesion on root surface which may optimize periodontal healing.

Compared morphology of the immatures of males of two urban ant species of camponotus

The immatures of males of two species of Camponotus ants (Hymenoptera: Formicidae) are described and compared by light and electron microscopy. The numbers of larval instars were determined: Camponotus rufipes Fabricius (Hymenoptera: Formicidae) have four instars; and Camponotus vittatus Forel have three. Male larvae of the two species are similar to previously described Camponotus larvae, sharing the following traits: basic shape of body and mandible, presence of 'chiloscleres', 'praesaepium' (some specimens), labial pseudopalps, and ten pairs of spiracles. However, larvae of the two species can be separated by bodily dimensions and based on their hair number and types. Worker larvae of C. vittatus previously described are extensively similar to male larvae, with only a few inconspicuous differences that may result from intraspecific variation or sexual differences.

Effects of ascorbic acid on in vitro culture of bovine preantral follicles

The objective of this study was to evaluate the effects of adding ascorbic acid to the media for in vitro culture of cattle ovarian fragments and to determine their effects on growth activation and viability of early-stage follicles. The ovarian cortex was divided into small fragments; one fragment was immediately fixed (control) and the other fragments were cultured in minimum essential medium (MEM) supplemented or not with various doses of ascorbic acid. Ovarian tissue was processed for histology, transmission electron microscopy (TEM) and immunohistochemical demonstration of proliferating cell nuclear antigen (PCNA). Compared with control fragments, the percentage of primordial follicles was reduced (p < 0.05) and the percentage of growing follicles had increased (p < 0.05) in cultured cortical fragments, independent of the tested medium or incubation time. Furthermore, compared with control tissue, culture of ovarian cortex for 8 days reduced the percentages of healthy, viable follicles (p < 0.05), but not when cultures were supplemented with 25, 50 or 100 μg/ml of ascorbic acid. Ultrastructural and immunohistochemical analysis of 8 day cultured ovarian cortical fragments, however, showed the integrity and viability of follicles only when fragments were cultured in presence of 50 μg/ml of ascorbic acid. In conclusion, this study demonstrated that addition of ascorbic acid to MEM at a concentration of 50 μg/ml not only stimulates the activation of 8 day in vitro cultured cattle primordial follicles and subsequent growth of activated follicles, but also safeguards the viability of these early-stage follicles. © 2012 Copyright Cambridge University Press.

Effects of treatment of the fat body trophocytes of Melipona quadrifasciata anthidioides nurse workers and virgin queens in culture by juvenile hormone III and ecdysterone (20-HE)

The fat body (FB) consists of two types of cells: throphocytes and oenocytes. Throphocytes are related to intermediary metabolism storing lipids, carbohydrates, and proteins while oenocytes play role in the lipids and lipoproteins production. The vitellogenin is the precursor of egg yolk (vitelline) and is synthesized on FB. The aim of this work was to analyze the effects of hormones acting in bee reproduction, as juvenile hormone (JH) and ecdisteroids (20 HE) on FB cells, where vitellogenin is synthesized. For the study were chose nurse workers that in Melipona quadrifasciata anthidioides present activated ovaries and produce eggs, and virgin queens whose ovaries are not yet activated, presenting only previtellogenic follicles. FB trophocytes from these classes of bees were cultivated in media containing different amounts of JH and 20-HE. The effects on trophocytes cytoplasm reserves of lipids, proteins, and activity of acid phosphatase were compared by observing preparations from cultured FB, treated and control, by transmission electron microscopy (TEM). The results showed that the hormones effects are related to the bee's caste and functional ovary stage. The role of acid phosphatase on mobilization of the trophocyte reserves was also determined. © 2012 Wiley Periodicals, Inc.

Bond strength of adhesive systems with different solvents to dry and wet dentin

Aim: This study evaluates bond strength between dentin and composite using adhesives with different solvents to dry and wet dentin. Materials and methods: Ninety bovine incisors were used; the vestibular surfaces were worn by the exposure of an area with a diameter of 4 mm of dentin. The specimens were divided into 6 groups, according to the type of adhesive used and hydratation stals: Group SB-wet: Single Bond 2 in wet dentin, Group SBdry: Single Bond 2 in dry dentin, Group SL-wet: Solobond M in wet dentin, Group SL-dry: Solobond M in dentin dry. Group XPwet: XP Bond in wet dentin, Group XP-dry: XP Bond in dentin dry. They were cut to obtain specimens in the shape of stick with 1 × 1 mm and subjected to microtensile test in universal testing machine with a cross speed of 1mm/min. The data were analyzed with ANOVA and Tukey's tests (5%). Results: ANOVA showed significant differences for surface treatment and interaction, but no difference was found for adhesive factor. The Tukey's test showed that the samples with wet dentin shown higher values of bond strength. Conclusion: The adhesive did not influence in the bond strength. The groups with wet dentin showed higher values of bond strength than groups with dry dentin.

Influence of CVD diamond tips and Er:YAG laser irradiation on bonding of different adhesive systems to dentin

Aim: The aim of this study was to compare the microtensile bond strength of three adhesive systems, using different methods of dentin preparation. Materials and methods: A hundred and eight bovine teeth were used. The dentin from buccal face was exposed and prepared with three different methods, divided in 3 groups: Group 1 (DT)- diamond tip on a high-speed handpiece; Group 2 (CVD)-CVD tip on a ultrasonic handpiece; Group 3 (LA)-Er: YAG laser. The teeth were divided into 3 subgroups, according adhesive systems used: Subgroup 1-Adper Single Bond Plus/3M ESPE (SB) total-etch adhesive; Subgroup 2-Adper Scotchbond SE/3M ESPE (AS) selfetching adhesive; Subgroup 3-Clearfil SE Bond/Kuraray (CS) selfetching adhesive. Blocks of composite (Filtek Z250-3M ESPE) 4 mm high were built up and specimens were stored in deionized water for 24 hours at 37°C. Serial mesiodistal and buccolingual cuts were made and stick-like specimens were obtained, with transversal section of 1.0 mm2. The samples were submitted to microtensile test at 1 mm/min and load of 10 kg in a universal testing machine. Data (MPa) were subjected to ANOVA and Tukey's tests (p < 0.05). Results and conclusion: Surface treatment with Diamond or CVD tips associated with Clearfil SE Bond adhesive produced significantly lower bond strength values compared to other groups. Surface treatment with Er: YAG laser associated with Single Bond Plus or Clearfil SE Bond adhesives and surface treatment with CVD tip associated with Adper Scotchbond SE adhesive produced significantly lower bond strength values compared to surface treatment with diamond or CVD tips associated with Single Bond Plus or Adper Scotchbond SE adhesives. Clinical significance: Interactions between laser and the CVD tip technologies and the different adhesive systems can produce a satisfactory bonding strength result, so that these associations may be beneficial and enhance the clinical outcomes.

Ultra-structural mapping of sugarcane bagasse after oxalic acid fiber expansion (OAFEX) and ethanol production by Candida shehatae and Saccharomyces cerevisiae

Background: Diminishing supplies of fossil fuels and oil spills are rousing to explore the alternative sources of energy that can be produced from non-food/feed-based substrates. Due to its abundance, sugarcane bagasse (SB) could be a model substrate for the second-generation biofuel cellulosic ethanol. However, the efficient bioconversion of SB remains a challenge for the commercial production of cellulosic ethanol. We hypothesized that oxalic-acid-mediated thermochemical pretreatment (OAFEX) would overcome the native recalcitrance of SB by enhancing the cellulase amenability toward the embedded cellulosic microfibrils. Results: OAFEX treatment revealed the solubilization of hemicellulose releasing sugars (12.56 g/l xylose and 1.85 g/l glucose), leaving cellulignin in an accessible form for enzymatic hydrolysis. The highest hydrolytic efficiency (66.51%) of cellulignin was achieved by enzymatic hydrolysis (Celluclast 1.5 L and Novozym 188). The ultrastructure characterization of SB using scanning electron microscopy (SEM), atomic force microscopy (AFM), Raman spectroscopy, Fourier transform-near infrared spectroscopy (FT-NIR), Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) revealed structural differences before and after OAFEX treatment with enzymatic hydrolysis. Furthermore, fermentation mediated by C. shehatae UFMG HM52.2 and S. cerevisiae 174 showed fuel ethanol production from detoxified acid (3.2 g/l, yield 0.353 g/g; 0.52 g/l, yield, 0.246 g/g) and enzymatic hydrolysates (4.83 g/l, yield, 0.28 g/g; 6.6 g/l, yield 0.46 g/g). Conclusions: OAFEX treatment revealed marked hemicellulose degradation, improving the cellulases ability to access the cellulignin and release fermentable sugars from the pretreated substrate. The ultrastructure of SB after OAFEX and enzymatic hydrolysis of cellulignin established thorough insights at the molecular level. © 2013 Chandel et al; licensee BioMed Central Ltd.

The reproductive cycle of the male sleep snake Sibynomorphus mikanii (Schlegel, 1837) from southeastern brazil

This study describes the male reproductive cycle of Sibynomorphus mikanii from southeastern Brazil considering macroscopic and microscopic variables. Spermatogenesis occurs during spring-summer (September-December) and spermiogenesis or maturation occurs in summer (December-February). The length and width of the kidney, the tubular diameter, and the epithelium height of the sexual segment of the kidney (SSK) are larger in summer-autumn (December-May). Histochemical reaction of the SSK [periodic acid-Schiff (PAS) and bromophenol blue (BB)] shows stronger results during summer-autumn, indicating an increase in the secretory activity of the granules. Testicular regression is observed in autumn and early winter (March-June) when a peak in the width of the ductus deferens occurs. The distal ductus deferens as well as the ampulla ductus deferentis exhibit secretory activities with positive reaction for PAS and BB. These results suggest that this secretion may nourish the spermatozoa while they are being stored in the ductus deferens. The increase in the Leydig cell nuclear diameter in association with SSK hypertrophy and the presence of sperm in the female indicate that the mating season occurs in autumn when testes begin to decrease their activity. The peak activity of Leydig cells and SSK exhibits an associated pattern with the mating season. However, spermatogenesis is dissociated of the copulation characterizing a complex reproductive cycle. At the individual level, S. mikanii males present a continuous cyclical reproductive pattern in the testes and kidneys (SSK), whereas at the populational level the reproductive pattern may be classified as seasonal semisynchronous. © 2012 Wiley Periodicals, Inc.

Rhipicephalus sanguineus (Acari: Ixodidae) female ticks exposed to castor oil (Ricinus communis): An ultrastructural overview

Tick control has been accomplished through the use of synthetic acaricides, which has created resistant individuals, as well as contaminating the environment and nontarget organisms. Substances of plant origin, such as oils and extracts of eucalyptus and neem leaves, have been researched as an alternative to replace the synthetic acaricides. Ricinoleic acid esters from castor oil have recently been shown as a promising alternative in eliminating bacterial contamination during ethanol fermentation, by acting as an effective biocide. The same positive results have been observed when these esters are added to the food given to tick-infested rabbits. This study tested the effect of these substance on the reproductive system of Rhipicephalus sanguineus females, added to rabbit food, more specifically on oogenesis. For this, four groups were established: four control groups (CG1, CG2, CG3, and CG4) and four treatment groups (TG1, TG2, TG3, and TG4) with one rabbit in each (New Zealand White), used as hosts. After full 4 days feeding (semi-engorgement), the females were collected and had their ovaries extracted. In this study, it was observed that R. sanguineus females exposed to esters had their ovaries modified, which was demonstrated through transmission electron microscopy techniques. The addition of ricinoleic esters to the diet of tick-infested rabbits revealed how toxic such substances are for the cytoplasmic organelles of oocytes and pedicel cells. These compounds can change the morphophysiology of germ and somatic cells, consequently influencing their viability and, therefore, confirming that the ricinoleic acid esters from castor oil are a promising substance in the control of R. sanguineus. © 2012 Springer-Verlag Berlin Heidelberg.

Toxicological and histopathological effects of hydramethylnon on Atta sexdens rubropilosa (Hymenoptera: Formicidae) workers

The leaf-cut ants are important agricultural pest, because they can cause intense defoliation in plants and destroy large areas cultivated. Although there are several works for the control of these insects by examining the toxicity of natural chemical compounds on various species of ants, few are focused on analyses of morphological changes caused in the affected organs. The aim of this study was to evaluate the effects of hydramethylnon on Atta sexdens rubropilosa workers through toxicological bioassays and morphological analysis of the post-pharyngeal glands, midgut, and Malpighian tubules of these ants. Hydramethylnon dissolved either in acetone (HA) or in a mixture of acetone and soy oil (HAO) was added to the artificial diet at a concentration of 200 μg/mL. The workers fed daily with the diet containing hydramethylnon showed higher mortality than the controls, especially when HAO was used. Moreover, light and electron microscopy revealed morphological alterations in the midgut and Malpighian tubules of workers treated with HA, whereas alterations of the post-pharyngeal glands were observed in the HAO-treated group. These results indicated that the presence of soy oil provided an alternate route for the ingestion of the formicide's active ingredient and corroborated previous studies that suggested a role for the post-pharyngeal glands in lipid metabolism. Our findings suggest that the oil may carry hydramethylnon to the gland lumen, resulting in lower quantity of the active ingredient in the intestinal lumen and Malpighian tubules that explains the lower degree of morphological alterations in these structures in the workers treated with HAO. These results may provide insight into the toxicological effects of hydramethylnon on leaf-cutting ants and the use of vegetable oil as an adjuvant in baits to control ants. © 2012 Elsevier Ltd.

Cellular responses in the Malpighian tubules of Scaptotrigona postica (Latreille, 1807) exposed to low doses of fipronil and boric acid

Studies of sub-lethal effects of pesticide residues on stingless bees are scarce and morphological analysis of organs would add information to toxicological analysis in order to clarify the continuous exposure of Scaptotrigona postica to insecticides. The aim of this study was to evaluate the morphology and histochemistry of the Malpighian tubules (excretory organ) of S. postica exposed to fipronil or boric acid to detect cellular responses that indicate toxicity or adaptative mechanisms to stress induced by exposure of worker bees to low doses of these chemical compounds. Newly emerged bees were submitted to toxicological bioassays and morphological analyses by optical microscopy and Transmission Electron Microscopy, as well as histochemical methods, were performed to detect proteins and glycoconjugates. Additionally, immunohistochemical detection of DNA fragmentation and HSP70 (70-kDa Heat shock protein) were performed to detect cell death and stress response, respectively. Statistical analysis, for the bioassays conducted with ingestion of contaminated diet with boric acid at 0.75% (w/w) or with fipronil at 0.1μg/kg of food, showed that the survival of bees that ingested the contaminated diets were significantly different to the survival rate presented by the control group (P<0.0001). Although some characteristics indicative of initiation of cell death were observed, the cells remained metabolically active in the processes of excretion and inactivation of chemical compounds. The data from this study reinforce the importance of research on sublethal effects of low doses of pesticides on bees in an attempt to assess a possible realistic dose and evaluate the risk assessment of stingless bee S. postica foraging in the vicinity of cultivated fields and/or in green urban areas. © 2013 Elsevier Ltd.