Thiago Lopes Rocha

Proteomic and histopathological response in the gills of Poecilia reticulata exposed to glyphosate-based herbicide

Glyphosate-based herbicides (GBH) are one of the most used herbicide nowadays, whilst there is growing concern over their impact on aquatic environment. Since data about the early proteomic response and toxic mechanisms of GBH in fish is very limited, the aim of this study was to investigate the early toxicity of GBH in the gills of guppies Poecilia reticulata using a proteomic approach associated with histopathological index. Median lethal concentration (LC50,96 h) was determined and LC50,96h values of guppies exposed to GBH were 3.6 ± 0.4 mg GLIL(-1). Using two-dimensional gel electrophoresis associated with mass spectrometry, 14 proteins regulated by GBH were identified, which are involved in different cell processes, as energy metabolism, regulation and maintenance of cytoskeleton, nucleic acid metabolism and stress response. Guppies exposed to GBH at 1.82 mg GLIL(-1) showed time-dependent histopathological response in different epithelial and muscle cell types. The histopathological indexes indicate that GBH cause regressive, vascular and progressive disorders in the gills of guppies. This study helped to unravel the molecular and tissue mechanisms associated with GBH toxicity, which are potential biomarkers for biomonitoring water pollution by herbicides.

Morphologic analysis of developmental phases and gill ontogenesis in neotropical species Poecilia vivipara (Cyprinodontiformes: Poeciliidae) exposed to different salinities

The present study evaluated and characterized the different phases of development and the cell types present in the gill epithelium of embryos, larvae, and fingerlings of guppy (Poecilia vivipara Bloch & Schneider, 1801) specimens exposed to water with salinity ranging from 0‰ to 20‰. The development of P. vivipara was classified into eight phases. The yolk was intensely eosinophilic, with glycidic and lipidic components between phases 1 to 7. After the development of their branchial epithelium, the presence of pavement cells, mitochondria-rich cells, and mucous cells (MCs) was observed. The understanding of ontogenesis indicates the sequence of cellular origin. Only type IV MCs located in the gill rays was detected from phase 3 through phase 7. Other types of MCs were observed only when the fingerlings were exposed to salinity changes and showed to be lectin-reactive, thus indicating that P. vivipara modulates the cellular behavior when submitted to salinity changes. Exposure to variable saline concentrations did not interfere in the initial development of guppies. The presence of a yolk sac until the fingerling phase characterized this species as ovoviviparous.

Chronic exposure to cannabidiol induces reproductive toxicity in male Swiss mice: MALE REPRODUCTIVE EFFECTS OF CBD

Children and adults with frequent and severe episodes of epilepsy that do not respond to standard treatments (such as carbamazepine, phenytoin and valproate) have long been prescribed cannabidiol (CBD) as an anticonvulsant drug. However, the safety of its chronic use in relation to reproduction has not been fully examined. This study aimed to assess the effects of chronic CBD exposure on the male reproductive system. CBD was orally administered to 21‐day‐old male Swiss mice at doses of 15 and 30 mg kg−1 daily (CBD 15 and 30 groups, respectively), with a control group receiving sunflower oil, for 34 consecutive days. After a 35 day recovery period, the following parameters were evaluated: weight of reproductive organs, testosterone concentration, spermatogenesis, histomorphometry, daily sperm production and its morphology. The CBD 30 group had a 76% decrease in total circulating testosterone, but it remained within the physiological normal range (240–1100 ng dl−1). CBD treatment induced a significant increase in the frequency of stages I–IV and V–VI of spermatogenesis, and a decrease in the frequency of stages VII–VIII and XII. A significant decrease in the number of Sertoli cells was observed only in the CBD 30 group. In both CBD groups the number of spermatozoa in the epididymis tail was reduced by 38%, sperm had head abnormalities, and cytoplasmic droplets were observed in the medial region of flagellum. These results indicated that chronic CBD exposure was associated with changes in the male reproductive system, suggesting its reproductive toxicity.

Análise do desenvolvimento e das glicoproteínas presentes nos folículos ovarianos de Poecilia vivipara (Cyprinodontiformes, Poeciliidae)

The morphofunctional aspects of oogenesis of Poecilia vivipara were studied aiming to understand the reproductive biology and development of species with internal fertilization, particularly those belonging to the family Poeciliidae. The stages of gonadal maturation and follicular development were characterized using mesoscopic, histological, histochemical, and lectin cytochemical analyses. Through mesoscopic evaluation the ovarian development was classified in six phases of development: immature, in maturation I, in maturation II, mature I, mature II, and post-spawn. Based on microscopic examination of the ovaries, we identified the presence of oocytes types I and II during the previtellogenic phase and types III, IV, and V during the vitellogenic phase. As oogenesis proceeded the oocyte cytosol increased in volume and presented increased cytoplasmic granule accumulation, characterizing vitellogenesis. The zona radiata (ZR) increased in thickness and complexity, and the follicular epithelium, which was initially thin and consisting of pavimentous cells, in type III oocytes exhibited cubic simple cells. The histochemical and cytochemical analyses revealed alterations in the composition of the molecular structures that form the ovarian follicle throughout the gonadal development. Our study demonstrated differences in the female reproductive system among fish species with internal and external fertilization and we suggest P. vivipara can be used as experimental model to test environmental toxicity.

Ovary histology and quantification of hemolymph proteins of Rhipicephalus (Boophilus)microplus treated with Melia azedarach

This study aimed to analyze ovary histology and quantify total protein in the hemolymph of Rhipicephalus (Boophilus) microplus females treated with hexane extracts from green fruits of Melia azedarach. Eight engorged females were immersed in the extract at 0.25% concentration, and eight in water containing 5% acetone (control). The females were dissected 72 hours after treatment, and the ovaries were weighed and subjected to standard histological techniques. The total protein concentration was measured in the hemolymph of 200 females, of which 100 were treated as described above and 100 served as a control. In the treated group, ovary weight reduction and predominance of immature oocytes were observed. In addition, there were decreases in the diameters of the cytoplasm and germ vesicle of the oocytes in the treated group, compared with the controls. The protein concentration in the hemolymph was higher in the treated group than in the controls. The morphological changes observed in the treated ovaries included: presence of vacuolization; alteration of oocyte morphology, which changed from rounded to elongated; deformation of the chorion; and disorganization of the yolk granules. These results demonstrate the action of M. azedarach fruit extracts on R. (B.) microplus oogenesis.

EDUCAÇÃO INCLUSIVA E A FORMAÇÃO DE PROFESSORES DE CIÊNCIAS: O PAPEL DAS UNIVERSIDADES FEDERAIS NA CAPACITAÇÃO DOS FUTUROS EDUCADORES

A inclusao de alunos com necessidades educacionais especiais (NEE’s) no sistema regular de ensino e hoje a diretriz principal das politicas publicas educacionais tanto a nivel federal, quanto estadual e municipal. Entretanto, os licenciados egressos das instituicoes de Ensino Superior estao verdadeiramente preparados para o atendimento adequado aos portadores de NEE’s? No sentido de responder este questionamento foi que demos inicio ao desenvolvimento dessa pesquisa que analisou as opinioes de academicos dos cursos de Fisica, Quimica e Biologia - Licenciaturas, do segundo e do ultimo periodo de cada curso, a fim de observar qual a reflexao dos mesmos em relacao a inclusao educacional e a ausencia de disciplinas desta area nos cursos de licenciatura nas universidades publicas federais.

Análise da Pluviosidade Mensal e Sazonal nas Bacias Hidrográficas do Ribeirão Santo Antônio e José Manoel no estado de Goiás

Este estudo teve como objetivo compreender a distribuicao da precipitacao pluvial, nas escalas mensal e sazonal, das bacias hidrograficas do Ribeirao Santo Antonio e Jose Manuel, localizadas no estado de Goias.xa0 Os dados pluviometricos foram disponibilizados pelo site da Agencia Nacional de Aguas (ANA), atraves do sistema hidroweb (2015). Foram baixados dados de chuva em 4 estacoes proximas a area de estudo, referentes ao periodo de 1974 a 2016. Os dados foram tabulados no programa Excel , extraindo a media mensal da serie historica analisada. Os resultados obtidos pela analise mensal de toda a serie historica mostraram que os meses de janeiro e dezembro foram os que apresentaram as maiores medias pluviometricas, com valores registrados acima dos 250mm, exceto para a estacao de Montividiu/GO, que registrou uma media abaixo de 240mm para toda a serie. Com isso, conclui-se que os meses com menores valores foram maio, junho, julho e agosto, obtendo media mensal de menos 30mm para a serie historica. Para a escala sazonal da area de estudo, o verao caracterizou-se como a estacao com maiores indices pluviometricos e o inverno a estacao mais seca.

In vitro activity of 3β-O-tigloylmelianol from Guarea kunthiana A. Juss (Meliaceae) on oogenesis and ecdysis of the cattle tick Rhipicephalus (Boophilus) microplus (Canestrini) (Acari: Ixodidae)

We evaluated the effects of 3β-O-tigloylmelianol from Guarea kunthiana A. Juss (Meliaceae) on oogenesis, as a larvicide and on ecdysis of the larvae and the nymphs of the cattle tick Rhipicephalus (Boophilus) microplus (Canestrini) (Acari: Ixodidae). On the oogenesis test, 48 engorged females were divided into three groups, evaluated at 24, 48 and 72 h post-treatment. Half of the females were treated with 0.01% 3β-O-tigloylmelianol diluted in distilled water and 5% dimethyl sulfoxide (DMSO), while the other half (controls) were exposed to distilled water and 5% DMSO. After treatment, the ovaries were weighed in order to measure the gonadosomatic index (GSI) and were also subjected to standard histological technical tests. On the larvicide and ecdysis tests, 3β-O-tigloylmelianol was tested at concentrations of 0.01, 0.005, 0.0025 and 0.00125%. Compared with the controls, there was a reduction of GSI of approximately 50% on the treated group, which started at 48 h post treatment. Overall, the protolimonoid 3β-O-tigloylmelianol has caused a significant reduction in the number of oocytes. It has also caused alteration of the cytoplasmic and germinal vesicle diameters. Morphological changes, such as vacuolization, chorion irregularity which has modified the oocytes morphology as well as alterations on the yolks granules were also observed. The compound was not larvicide, however, interfered in the ecdysis of the larvae and the nymphs. This study shows that the protolimonoid 3β-O-tigloylmelianol from G. kunthiana acts on oogenesis and ecdysis of R. (B.) microplus, but not as larvicide, indicating that it acts on the endocrine system of the tick.

Pseudobrânquia do guaru Poecilia reticulata (Peter, 1859): análise estrutural, morfométrica e histoquímica para detecção de glicoconjugados

The morphology, cytomorphometric parameters, and glycoconjugates present in the pseudo-gill of guppy, Poecilia reticulata Peter, 1859 (Cyprinodontiformes: Poeciliidae), were investigated by light microscopy coupled to image capture and analysis system, and also by lectin histochemistry. The microscopic anatomy indicates that P. reticulata has a glandular pseudo-gill formed by two lobes, located underneath the pharynx epithelium. The organ is formed by vascularized parenchyma rich in pseudo-gill cells. This cell type exhibits active cytophysiological state with an abundant system of biomembranes and lacking of ostium in apical surface, which in turn is found in the mitochondria-rich cells of the holobranch. This indicates that the pseudo-gill cells distinguishe from the holobranch cells in their morphology, histochemistry and physiology. Due to these intrinsic characteristics, the pseudo-gill of guppy fingerlings may have non-respiratory function in the initial phase of their development. The characterization of guppys pseudo-gill could facilitate further studies about the effect of water pollutants on biomonitor species, such as P. reticulata.