Oxidative stress, biotransformation enzymes and histopathological alterations in Nile tilapia (Oreochromis niloticus) exposed to new and used automotive lubricant oil.

Lubricant oils are among oil-based products that are not fully consumed during its use, thereby producing non-biodegradable residues which can cause contamination of natural systems. This study evaluated the toxicity of new and used lubricating oil (0.01 and 0.1 mL L-1) in adult Nile tilapia (Oreochromis niloticus), by assessing the effects on oxidative stress, biotransformation enzymes (liver and gills), and histopathological alterations on hepatic and pancreatic tissues after 3 and 7 days of exposure. Results showed that 3-days exposure to 0.1 mL L-1 of used and new lubricating oil increased the activity of superoxide dismutase (SOD) and malondialdehyde (MDA) levels in liver of O. niloticus, respectively. In gills, catalase (CAT) was decreased in fish exposed to 0.1 mL L-1 of non-used oil after 3 days, but pronounced increases in CAT was detected after 7 days-exposure to both new and used oil. Shorter exposure to both concentrations of new and used oil also raised glutathione-S-transferase activity (GST) in gills. Ethoxyresorufin-O-deethylase (EROD) was induced in liver of fish exposed to 0.1 mL L-1of used oil after 3 and 7 days, however a reduced response of this enzyme was detected in gills of animals from both oil treatments. In vitro analysis showed that hepatic EROD was inhibited by lubricating oil exposures, with more pronounced responses in treatments containing used oil. Hepatic lesions, such as cytoplasmic vacuolization, nuclei abnormally, changes in hepatocytes shape, steatosis, cholestasis, eosinophilic inclusions and necrosis were mainly increased by 7 days exposure to used lubricating oil at higher concentration.

Variação morfológica no padrão lobar e nas fissuras dos pulmões: relato de caso / Morphological variation in the lobar pattern and in the fissures of the lungs: a case report

Modelo do estudo: Relato de caso. Importância do problema: Lobos e fissuras acessórias nos pulmões podem induzir a erros de interpretação e diagnóstico, além de favorecer a disseminação de patologias como a pneumonia para lobos adjacentes. Dessa forma, a constatação dessas alterações anatômicas pode colaborar em diagnósticos clínicos e procedimentos cirúrgicos. Comentários: Relatamos um caso raro de modificação anatômica nos lobos e fis-suras dos pulmões. O pulmão esquerdo exibia um lobo médio acessório e uma fissura horizontal completa. Por outro lado, o pulmão direito apresentava duas fissuras acessórias incompletas situadas inferiormente na face costal do lobo médio. Além disso, observamos que o hilo do pulmão esquerdo continha dois brônquios lobares (superior e inferior), um brônquio lobar acessório e três artérias pulmonares. O conhecimento das variações anatômicas aqui relatadas é fundamental, pois auxiliam os profissionais da saúde nos diagnósticos e nas decisões terapêuticas e cirúrgicas.(AU)

Study: Case Report. Importance: Lobos and accessory fissures in the lungs can induce errors of interpretation and diagnosis, as well as favor the spread of pathologies such as pneumonia to adjacent lobes. Thus, the confirmation of these anatomical alterations can collaborate in clinical diagnoses and surgical procedures. Comments: We report a rare case of anatomical variation in the lobes and fissures of the lungs. The left lung showed an accessory middle lobe and a complete horizontal fissure. On the other hand, the right lung had two incomplete accessory fissures located inferiorly in the costal face of the middle lobe. In addition, we observed that the left lung hilum contained two lobar bronchi (upper and lower), one accessory lobar bronchus and three pulmonary arteries. Knowledge of the anatomical variations reported here is fundamental, as they help health professionals in diagnoses and therapeutic and surgical decisions (AU)

Effects of methylphenidate on the aggressive behavior, serotonin and dopamine levels, and dopamine-related gene transcription in brain of male Nile tilapia (Oreochromis niloticus).

The occurrence of pharmaceuticals in the aquatic environment has increased considerably in the last decades, causing negative biochemical, physiological, and behavioral effects in aquatic organisms. In this study, we evaluated the effects of methylphenidate (MPH) on the aggressive behavior, dopamine-related gene transcript levels, monoamine levels, and carboxylesterase transcript levels and activity in the brain of male Nile tilapia (Oreochromis niloticus). Carboxylesterase activity was also measured in the liver and gills. Fish were exposed for 5 days to MPH at 20 and 100 ng L-1. Fish exposed to 100 ng L-1 of MPH showed increased aggressiveness and decreased dopamine (DA) and serotonin (5-HT) levels. No changes were observed in plasma testosterone levels and in the transcript levels of D1 and D2 dopamine receptors, dopamine transporter (DAT), and carboxylesterase 2 (CES2). Exposure to 100 ng L-1 of MPH caused a decrease in the transcript levels of carboxylesterase 3 (CES3) and an increase in tyrosine hydroxylase (TH), while exposure to 20 ng L-1 of MPH increased the transcript levels of D5 dopamine receptor. Carboxylesterase activity was unchanged in the brain and liver and increased in the gills of fish exposed to 20 ng L-1. These results indicate that MPH at 100 ng L-1 increases aggressiveness in Nile tilapia, possibly due to a decrease in 5-HT levels in the brain and alterations in dopamine levels and dopamine-related genes.

Anatomical characterization of the digestive system of the pufferfish (Chilomycterus spinosus spinosus)

Descriptive studies of the fish digestive system are fundamental because they provide information on the biology of the species. Thus, the objective of this study was to morphologically describe the digestive system of the pufferfish, Chilomycterus spinosus spinosus. For this, adult specimens of pufferfish (n = 10) of both sexes were used. The animals were fixed with 10% aqueous formaldehyde solution, dissected, analyzed descriptively and photographed. The results demonstrate that the pufferfish has a morphologically modified digestive system, which is adapted to the defense behavior. This species presents a pouch-shaped diverticulum, that is called abdominal pouch, which allows the expansion of the celomatic cavity and the temporary storage of food. Although it is used to store food, macroscopically the abdominal pouch does not show gastric folds. However, this absence is compensated by a small intestine containing innumerable villi.

Anatomical description of the salivary glands: parotid, mandibular and sublingual of the crab-eating-fox (Cerdocyon thous) / Descrição anatômica das glândulas salivares: parótida, mandibular e sublingual do cachorro-do-mato (Cerdocyon thous)

Descriptive anatomical studies of wild animals are fundamental, since they providesubsidies for the elaboration of more adequate techniques of sustained management, contributing to thepreservation of species threatened with extinction. Thus, the aim of the study is to anatomically describethe salivary glands: parotid, mandibular and sublingual of the crab-eating-fox (Cerdocyon thous). For thispurpose, three carcasses of crab-eating-fox donated by Ibama were used. The animals were fixed with 10%aqueous formaldehyde solution, dissected, analyzed descriptively and photographed. It was found that theparotid gland of the crab-eating-fox is located caudally to the branch of the mandible, in the rostroventralmargin of the auricular cartilage. The mandibular gland is located in the atlantal fossa and extends to thebasi-hyoid bone. The sublingual gland of the crab-eating-fox shows two parts: monostomatic andpolystomatic. The monostomatics are located in the occipitomandibular region of the digastric muscle andthe portion polysomatic is situated between the tunica mucosa of the oral cavity and the buccinator muscle.Based on the results, we conclude that the salivary glands of the crab-eating-fox are found distributed inthe facial region and present anatomical characteristics that follow the same structural pattern described forother species of carnivorous mammals.

Estudos anatômicos descritivos de animais silvestres são fundamentais, pois fornecemsubsídios para a elaboração de técnicas mais adequadas de manejo sustentado, contribuindo com apreservação de espécies ameaçadas de extinção. Dessa forma, o objetivo do estudo é descreveranatomicamente as glândulas salivares: parótida, mandibular e sublingual do cachorro-do-mato (Cerdocyonthous). Para isso, foram utilizadas três carcaças de cachorro-do-mato doados pelo Ibama. Os animais foramfixados com solução aquosa de formol a 10%, dissecados, analisados descritivamente e fotografados.Constatou-se que a glândula parótida do cachorro-do-mato situa-se caudalmente ao ramo da mandíbula,disposta na margem rostroventral da cartilagem auricular. A glândula mandibular está situada na fossaatlantal e estende-se até o osso basi-hióide. A glândula sublingual do cachorro-do-mato apresenta duasporções: monostomática e polistomática. A monostomática situa-se na região occiptomandibular domúsculo digástrico e a polistomática entre a túnica mucosa da cavidade oral e o músculo bucinador. Combase nos resultados concluímos que as glândulas salivares do cachorro-do-mato são encontradasdistribuídas pela região facial e apresentam características anatômicas que seguem o mesmo padrãoestrutural descrito para outras espécies de mamíferos carnívoros.

Leporinus elongatus induced spawning using carp pituitary extract or mammalian GnRH analogue combined with dopamine receptor antagonists.

Several studies have been developed to support the replacement of the crude carp pituitary extract (CPE) by synthetic products for induced reproduction of South American rheophilic species. However, results have been quite heterogeneous and there is no consensus or a routine use of synthetic products in these species. Thus, the aim of this study was to evaluate the ovulatory process in L. elongatus using different protocols of hormonal induction. Thus, fifteen wild mature females maintained at the Experimental Fish Station, Salto Grande, SP, Brazil were submitted to three different hormonal treatments: CPE (fractioned dose: 0.5 and 5.0 mg kg-1); mGnRHa (single dose: 3.5 µg kg-1) and mGnRHa (single dose: 5.0 µg kg-1). The spawning rate and absolute fecundity were similar among the treatments, but fertility rates were higher for CPE treatment (23.60 ± 9.40) then for mGnRHa treatments (close to or zero zero). Although females ovulated in all treatments, none of them provided viable embryos, showing hatching rates close to zero or zero. Both mGnRHa treatments were more potent for inducing the ovulatory process then CPE treatment, which was evidenced by the fact that the formers showed higher volume density of postovulatory follicles (POF). Accordingly, E2 and 17α-OHP plasma levels were higher for the mGnRHa treated females compared to the CPE one at the time of ovulation. In this study we confirmed previous scientific evidence that, regardless of whether promoting ovulation, the use of conventional CPE and GnRH doses are not appropriate for some South American migratory species, due to the non-attainment of viable embryos. Moreover, we have brought new information about the relationship between reproductive performance and gonadal steroids concentrations using different hormonal therapies, contributing to understand the reasons for Leporinus elongatus embryo loss in induced spawning.

Estrogenic and anti-androgenic effects of the herbicide tebuthiuron in male Nile tilapia (Oreochromis niloticus).

Tebuthiuron is a phenylurea herbicide widely used in agriculture that can reach the aquatic environments, possibly posing negative effects to the aquatic biota. Phenylurea herbicides, such as diuron, are known to cause estrogenic and anti-androgenic effects in fish, but no such effects were yet reported for tebuthiuron exposure. Thus, the aim of this study was to evaluate if tebuthiuron, at environmentally relevant concentrations (100 and 200ng/L) and after 25days of exposure have estrogenic and/or anti-androgenic effects on male of Nile tilapia (Oreochromis niloticus), through the evaluation of plasmatic testosterone (T) and estradiol (E2) levels, brain aromatase (CYP19) levels (western-blot), and by evaluating the histology of the testicles. When compared to the control group, plasmatic T levels decreased about 76% in the animals exposed to 200ng/L of tebuthiuron, while E2 levels increased about 94%, which could be related to a significant increase (77%) in CYP19A1 levels, an enzyme that catalyzes the conversion of androgens into estrogens. Histological analyses of the testicles also demonstrated that tebuthiuron at both tested concentrations caused a decrease in the diameter of the seminiferous tubules and in the diameter of the lumen. Therefore, the gonadosomatic index (GSI) was reduced by 36% % in the animals exposed 200ng/L to tebuthiuron. Indeed, the relative frequency of spermatocytes and spermatids increased respectively 73% (200ng/L) and 61% (100ng/L) in the tebuthiuron exposed animals, possibly due to the impairment of sperm release into the lumen, that was decreased 93% (200ng/L) in the treated animals compared to the control. These results confirm that tebuthiuron causes estrogenic and anti-androgenic effects in Nile tilapias at environmentally relevant concentrations.

Diuron metabolites act as endocrine disruptors and alter aggressive behavior in Nile tilapia (Oreochromis niloticus).

Diuron and its biodegradation metabolites were recently reported to cause alterations in plasma steroid hormone concentrations with subsequent impacts on reproductive development in fish. Since steroid hormone biosynthesis is regulated through neurotransmission of the central nervous system (CNS), studies were conducted to determine whether neurotransmitters that control hormone biosynthesis could be affected after diuron and diuron metabolites treatment. As the same neurotransmitters and steroid hormones regulate behavioral outcomes, aggression was also evaluated in male Nile tilapia (Oreochromis niloticus). Male tilapias were exposed for 10 days to waterborne diuron and the metabolites 3,4-dichloroaniline (DCA), 3,4-dichlorophenyl-N-methylurea (DCPMU), at nominal concentrations of 100 ng L-1. In contrast to Diuron, DCA and DCPMU significantly diminished plasma testosterone concentrations (39.4% and 36.8%, respectively) and reduced dopamine levels in the brain (47.1% and 44.2%, respectively). In addition, concentrations of the stress steroid, cortisol were increased after DCA (71.0%) and DCPMU (57.8-%) exposure. A significant decrease in aggressive behavior was also observed in animals treated with the metabolites DCA (50.9%) and DCPMU (68.8%). These results indicate that biotransformation of diuron to active metabolites alter signaling pathways of the CNS which may impact androgen and the stress response as well as behavior necessary for social dominance, growth, and reproduction.

Estrogenic activities of diuron metabolites in female Nile tilapia (Oreochromis niloticus).

Some endocrine disrupting chemicals (EDCs) can alter the estrogenic activities of the organism by directly interacting with estrogen receptors (ER) or indirectly through the hypothalamus-pituitary-gonadal axis. Recent studies in male Nile tilapia (Oreochromis niloticus) indicated that diuron may have anti-androgenic activity augmented by biotransformation. In this study, the effects of diuron and three of its metabolites were evaluated in female tilapia. Sexually mature female fish were exposed for 25 days to diuron, as well as to its metabolites 3,4-dichloroaniline (DCA), 3,4-dichlorophenylurea (DCPU) and 3,4-dichlorophenyl-N-methylurea (DCPMU), at concentrations of 100 ng/L. Diuron metabolites caused increases in E2 plasma levels, gonadosomatic indices and in the percentage of final vitellogenic oocytes. Moreover, diuron and its metabolites caused a decrease in germinative cells. Significant differences in plasma concentrations of the estrogen precursor and gonadal regulator17α-hydroxyprogesterone (17α-OHP) were not observed. These results show that diuron metabolites had estrogenic effects potentially mediated through enhanced estradiol biosynthesis and accelerated the ovarian development of O. niloticus females.

Anti-androgenic activities of diuron and its metabolites in male Nile tilapia (Oreochromis niloticus).

Diuron (3-(3,4-dichlorophenyl)-1,1-dimethylurea) is a widely used herbicide which has been frequently detected in surface waters throughout the world. In vivo bioassay guided fractionation studies indicated that diuron may have estrogenic activity augmented by biotransformation. This study evaluated the effects of diuron and three of its metabolites on plasma hormone concentrations and spermatogenesis of the freshwater fish Nile tilapia (Oreochromis niloticus). Sexually mature male fish were exposed for 25 days to diuron, as well to its metabolites 3,4-dichloroaniline (DCA), 3,4-dichlorophenylurea (DCPU) and 3,4-dichlorophenyl-N-methylurea (DCPMU), at concentrations of 200ng/L. Testosterone levels were decreased by diuron, but had limited effects on gonadal histology. Diuron metabolites, however, caused significant decreases in testosterone and in 11-ketotestosterone, gonadosomatic index, diameter of seminiferous tubules and in the mean percentages of germ cells (spermatids and spermatozoa). We conclude that these metabolites have antiandrogenic activity to male Nile tilapia, potentially causing reproductive impairment in male fish.

Dynamics of ovarian maturation during the reproductive cycle of Metynnis maculatus, a reservoir invasive fish species (Teleostei: Characiformes)

In this study, we evaluated the dynamics of ovarian maturation and the spawning processes during the reproductive cycle of Metynnis maculatus. Adult females (n = 36) were collected bimonthly between April 2010 and March 2011. The mean gonadosomatic index (GSI) was determined, ovarian and blood samples were submitted for morphometric evaluation and the steroid plasma concentration was determined by ELISA. This species demonstrated asynchronous ovarian development with multiple spawns. This study revealed that, although defined as a multiple spawning species, the ovaries of M. maculatus have a pattern of development with a predominance of vitellogenesis between April and August and have an intensification in spawning in September; in October, a drop in the mean GSI values occurred, and the highest frequencies of post-ovulatory follicles (POFs) were observed. We observed a positive correlation between the POF and the levels of 17α-hydroxyprogesterone. Metynnis maculatus has the potential to be used as a source of pituitary tissue for the preparation of crude extracts for hormonal induction; the theoretical period for use is from September to December, but specific studies to determine the feasibility of this approach must be conducted.

Neste estudo, avaliamos a dinâmica da maturação ovariana a desova durante o ciclo reprodutivo de Metynnis maculatus. Fêmeas adultas (n = 36) foram coletadas bimestralmente entre abril de 2010 e março de 2011. O índice gonadossomático (IGS) foi calculado e amostras de ovário e de sangue foram submetidas à avaliação morfométrica e das concentrações plasmáticas dos esteroides por ELISA, respectivamente. A espécie apresenta desenvolvimento ovariano assincrônico, com múltiplas desovas. Neste estudo revelamos que mesmo sendo de desova parcelada, os ovários do M. maculatus mostraram um padrão de desenvolvimento com predomínio de atividade vitelogênica entre abril a agosto e intensificação da desova em setembro. Em outubro houve uma diminuição nos valores médios de IGS, bem como registramos as maiores frequências de folículos pós-ovulatórios (FPOs). Observamos uma correlação positiva entre a frequência de FPOs e a concentração plasmática de 17 α-OHP. O M. maculatus tem potencial para ser usado como fonte para uso de hipófise para preparo de extrato bruto para indução hormonal, sendo o período teórico para coleta de hipófises de setembro a outubro, mas estudos específicos para esta finalidade ainda precisam ser desenvolvidos.