Análises morfológica e histoquímicas do ventrículo e do cérebro de operárias recém emergidas de Apis Mellifera L. (Hymenoptera, Apidae) tratadas com Thiamethoxam

Apis mellifera honeybees are social insects of economic importance, by providing honeybee products, and by the pollination of natural areas of vegetation or agricultural areas. The constant use of pesticides, including the thiamethoxam, which is an insecticide belonging to the class of neonicotinoids with neurotoxic action, is subjecting pollinators to situations of severe stress, which has been evidenced by the decrease in the density of honeybees in many parts of the world. By these considerations, the present study aimed to assess the acute toxicity of thiamethoxam for newly emerged workers of A. mellifera, and to investigate the effect of sublethal doses of this insecticide on the survival time and its cytotoxicity to the brain and midgut to the honeybees. It was established the value of the lethal concentration 50 (LC50) equal to 4.28 ng thiamethoxam/μL of food and from it, the sublethal concentrations of 0.428 ng thiamethoxam/μL (CL50/10) and 0.0428 ng thiamethoxam/μL (CL50/100), which were used in bioassays of intoxication of the honeybees. After the bioassays the bees were dissected and the brain and midguts were collected to analyze possible morphological (staining with Hematoxylin-Eosin) and histochemical alterations (Xylidine Ponceau technique, and Feulgen and PAS reactions) caused by exposure to thiamethoxam, and to calculate the lethal time (LT50) for the workers. The duration of the bioassays was 8 days after beginning of feeding. The results obtained showed that the thiamethoxam is toxic to newly-emerged workers of A. mellifera, causing changes in survival time of individuals. This study also shown that the thiamethoxam causes morphological and histochemical alterations on the midgut and brain of workers. These alterations may be reflected in physiological and behavioral changes that can modify the operation of the colony

Cytochemical study of Rhodnius neglectus and Rhodnius prolixus salivary gland cells (Hemiptera, Triatominae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Análise da frequência de micronúcleos frente á utilização de colutório bucal com base alcoólica em indivíduos fumantes e não fumantes

Micronuclei (MN) originate from chromosome fragments or whole chromosomes that lag behind at anaphase during cell division can occur due to excessive exposure to harmful agents, defects in mitosis and / or DNA repair process. The study evaluated the frequency of micronuclei in cytological samples after daily use of alcohol -based mouthwash mouth in smokers and non-smokers. We assessed 30 subjects, aged between 17-42 years and established as exclusion criteria: alcohol consumption; periodontal disease; systemic diseases that may interfere with the integrity of the oral mucosa; individuals with medical or pharmacological history that affect the performance of the study, family history of malignancies. For smokers the inclusion criterion was a minimum consumption of 20 cigarettes / day, and the contrast required for non-smokers. The sample was divided into 02 experimental groups: Group 1 Smokers and Group 2 Non-Smokers, both groups performed mouthwash (alcohol 21,6%) with Listerine® Defense of Teeth and Gums for 04 weeks cytological collection were performed on buccal mucosa and portion median border of the tongue, one for initial, week 0 control, and subsequent three weeks with continued use of mouthwash. The collected material was stained by specific staining Feulgen / Fast Green. The analysis of 3000 cells / individual was performed by light microscopy statistics used Kruskal- Wallis and Mann-Whitney tests for frequency MN. The frequency of micronuclei in buccal mucosa was 1.6 ± 1.75 and 1.46 ± 2.06 for the site language to Group 1 Smokers of showing a slight increase compared with Group 2 Non-smoking, which showed a variation of 1.2 ± 1.53 for the buccal mucosa site and 0.86 ± 1.41 for the site language. However, this difference was not statistically significant. The results suggest that alcohol-based mouthwash tested is not an inducer of genotoxic changes, thus not changing the frequency...

A step to the gigantic genome of the desert locust: chromosome sizes and repeated DNAs

The desert locust (Schistocerca gregaria) has been used as material for numerous cytogenetic studies. Its genome size is estimated to be 8.55 Gb of DNA comprised in 11 autosomes and the X chromosome. Its X0/XX sex chromosome determinism therefore results in females having 24 chromosomes whereas males have 23. Surprisingly, little is known about the DNA content of this locust's huge chromosomes. Here, we use the Feulgen Image Analysis Densitometry and C-banding techniques to respectively estimate the DNA quantity and heterochromatin content of each chromosome. We also identify three satellite DNAs using both restriction endonucleases and next-generation sequencing. We then use fluorescent in situ hybridization to determine the chromosomal location of these satellite DNAs as well as that of six tandem repeat DNA gene families. The combination of the results obtained in this work allows distinguishing between the different chromosomes not only by size, but also by the kind of repetitive DNAs that they contain. The recent publication of the draft genome of the migratory locust (Locusta migratoria), the largest animal genome hitherto sequenced, invites for sequencing even larger genomes. S. gregaria is a pest that causes high economic losses. It is thus among the primary candidates for genome sequencing. But this species genome is about 50 % larger than that of L. migratoria, and although next-generation sequencing currently allows sequencing large genomes, sequencing it would mean a greater challenge. The chromosome sizes and markers provided here should not only help planning the sequencing project and guide the assembly but would also facilitate assigning assembled linkage groups to actual chromosomes.

Transdifferentiation of Mature Cortical Cells to Functional Abscission Cells in Bean1

Abscission explants of bean (Phaseolus vulgaris L.) were treated with ethylene to induce cell separation at the primary abscission zone. After several days of further incubation of the remaining petiole in endogenously produced ethylene, the distal two-thirds of the petiole became senescent, and the remaining (proximal) portion stayed green. Cell-to-cell separation (secondary abscission) takes place precisely at the interface between the senescing yellow and the enlarging green cells. The expression of the abscission-associated isoform of β-1,4-glucanhydrolase, the activation of the Golgi apparatus, and enhanced vesicle formation occurred only in the enlarging cortical cells on the green side. These changes were indistinguishable from those that occur in normal abscission cells and confirm the conversion of the cortical cells to abscission-type cells. Secondary abscission cells were also induced by applying auxin to the exposed primary abscission surface after the pulvinus was shed, provided ethylene was added. Then, the orientation of development of green and yellow tissue was reversed; the distal tissue remained green and the proximal tissue yellowed. Nevertheless, separation still occurred at the junction between green and yellow cells and, again, it was one to two cell layers of the green side that enlarged and separated from their senescing neighbors. Evaluation of Feulgen-stained tissue establishes that, although nuclear changes occur, the conversion of the cortical cell to an abscission zone cell is a true transdifferentiation event, occurring in the absence of cell division.

Perpetuation of the agent of human granulocytic ehrlichiosis in a deer tick-rodent cycle.

A human-derived strain of the agent of human granulocytic ehrlichiosis, a recently described emerging rickettsial disease, has been established by serial blood passage in mouse hosts. Larval deer ticks acquired infection by feeding upon such mice and efficiently transmitted the ehrlichiae after molting to nymphs, thereby demonstrating vector competence. The agent was detected by demonstrating Feulgen-positive inclusions in the salivary glands of the experimentally infected ticks and from field-derived adult deer ticks. White-footed mice from a field site infected laboratory-reared ticks with the agent of human granulocytic ehrlichiosis, suggesting that these rodents serve as reservoirs for ehrlichiae as well as for Lyme disease spirochetes and the piroplasm that causes human babesiosis. About 10% of host-seeking deer ticks were infected with ehrlichiae, and of these, 20% also contained spirochetes. Cotransmission of diverse pathogens by the aggressively human-biting deer tick may have a unique impact on public health in certain endemic sites.

Estudio de la muerte celular por Apoptosis en el ovario de Columba maculosa y Columbina picui (Aves:Columbidae) durante el Ciclo Reproductivo Anual.”

En este trabajo se analizarán las características estructurales, cuantitativas y el proceso de muerte celular por apoptosis en el ovario de C. maculosa y C.picui con el objetivo de aportar conocimientos básicos a la biología reproductiva de estas aves. Cincuenta hembras adultas de cada especie se capturarán en el Dpto. Río Primero (Pcia. de Cba), R. Argentina, durante el ciclo reproductivo 2011-2012. Las muestras de ovarios se fijaran en Formalina Neutra pH 7.0, procesarán con la técnica de inclusión en parafina y colorearan con Hematoxilina /Eosina y Reacción Nuclear de Feulgen. Cinco muestras serán utilizadas para la determinación de muerte celular por apoptosis con la técnica de TUNEL. Se estudiarán las características morfohistológicas del ovario de C.maculosa y C. picui e identificarán, categorizarán y cuantificarán los folículos atrésicos no bursting (folículos atrésicos previtelogénicos y vcitelogénicos pequeños que conservan la integridad de la pared folicular) y bursting (folículos vitelogénicos mayores de 2 mm que liberar el contenido folicular por ruptura de la pared folicular) durante el ciclo reproductivo anual. Mediante la marcación de ADN fragmentado, se revelará la muerte celular por apoptosis en las células granulosas de los folículos atrésicos. Se compararán las semejanzas y diferencias estructurales y cuantitativas y el proceso de muerte celular en los folículos regresivos entre las dos especies. Los resultados de este trabajo representarán un importante aporte al conocimiento de la atresia folicular como así también a la muerte celular, un proceso estrechamente asociado a la misma, aún poco estudiado en el ovario de las aves.

Nuclear alterations in nasal mucosa epithelial cells of students exposed to formaldehyde

Introduction: Formaldehyde is a compound with a wide range and is commonly used in anatomy and pathology laboratories. At room temperature is quickly volatilized to a pungent and suffocating gas and its inhalation has been correlated to nuclear alterations in different tissues. We aimed to investigate whether exposure to this compound was correlated with the appearance of cytotoxic and genotoxic features in the nasal epithelial cells of students enrolled in a human anatomy course. Material and Methods: This prospective study collected periodically nasal cells from mucosa of 17 volunteers from two different undergraduate programs with different workloads of practical lessons in an anatomy laboratory, 30 and 90 hours per semester. Cells were staining according to Feulgen method and nuclear morphology was analyzed to detect possible damage. Dunn's post hoc test was used in the statistical analysis. Pearson's correlation was performed for gender, age and questionnaire responses. Results: Epithelial cells showed indicators of cytotoxicity and mutagenicity. Students with a more extensive workload in anatomy laboratory displayed a more severe profile with an increase in karyorrhexis (p < 0.05) over time. The micronucleus analysis showed difference between first and second collection (p < 0.01), although it was not maintained over the time. Students with a less extensive workload display no differences in most of cytological features. Despite karyorrhexis was present in a greater number of cells, for this group no significant difference was observed between any range. The same was observed to karyolysis and micronucleus (p > 0.05). Conclusion: Individuals exposed for short periods of time to formaldehyde are subject to the toxic action of this gas. Karyorrhexis was the most frequently observed cytotoxic feature and micronucleus showed an increase between the first time point. The patterns observed between the student's groups suggest a negative effect due to exposure time.