Oxidative activity of corpus luteum and ovarian parenchyma in Bos taurus indicus heifers

Abstract The aim of this study was to evaluate the oxidative stress in ovaries and corpus luteum (CL) of Bos taurus indicus females and the oxidant effect of CL in ovarian tissues in regions near, intermediate, or distant from it. Ovaries (n=12) of Nelore heifers (n=6) were collected from a slaughterhouse and fragmented. Experiment 1, each ovary was obtained from three fragments, resulting in 18 fragments of ovaries with CL (OV+CL) and another 18 fragments of ovaries without CL (OV-CL). Three fragments were generated from CL, totaling 18 CL fragments. In experiment 2, the ovarian fragments were removed from specific regions near, intermediate, or distant from the CL. All the fragments were placed in Eppendorf-type microtubes (1 mL), kept in a thermal container at 4 ºC, and then stored in a -80 ºC freezer for analysis of oxidative stress (TBARS and NBT) and antioxidant potential (FRAP and ABTS). In the antioxidant activity analysis, luteal tissues showed more antioxidant activity than ovarian tissue (FRAP = P < 0.0001; ABTS = P < 0.02). In the oxidative stress analysis, CL had lower levels of reactive oxygen species (ROS; TBARS = P < 0.03; NBT = P < 0.0001) than ovarian tissues. There was no difference in antioxidant activity and oxidative stress between the fragments obtained from different regions (OV+CL versus OV-CL; P > 0.05). The presence of CL in the ovaries of Bos taurus indicus females did not influence the oxidative stress or antioxidant potential of the gonad. Thus, the removal of ovarian fragments with or without the presence of CL indicates that biotechnologies such as in vitro follicle cultivation is possible.

Gonadal characterization of the Amazonian fish Serrapinnus kriegi (Characidae: Cheirodontinae) / Caracterização gonadal do peixe Amazônico Serrapinnus kriegi (Characidae: Cheirodontinae)

Abstract The knowledge of the testicular and ovarian morphology of a particular fish species is of paramount importance. Such analyze enables the development of studies and techniques aiming the improvement of their reproduction, management, commercialization and even their conservation. This study performed the ovarian and testicular characterization of the ornamental Amazon fish Serrapinnus kriegi. A total of three males and three females had their gonads analyzed by optical microscopy. Females present ovaries filled with oocytes in asynchronous development, indicating partial spawning in the species. Moreover, the micropyle and micropilar cell formation was observed in primary growing oocytes, representing a precocious oocyte development; and the zona radiata in the final vitellogenic oocytes is thicker than other related species, evidencing the development of a better protection to the embryos in function of the waters' turbulence that characterize it spawning sites in the Amazonian streams. The male specimens' present anastomosed tubular testes with unrestricted spermatogonia spread along the entire seminiferous tubules. The present data elucidate the dynamic of spermatogenesis and oogenesis of an ornamental Amazonian species, through the description of the male and female germ cells development.

Resumo O conhecimento da morfologia testicular e ovariana de uma determinada espécie de peixe é de suma importância, pois através destas análises é possível o desenvolvimento de estudos e técnicas visando o melhoramento de sua reprodução, manejo e comercialização e até mesmo auxiliar em sua conservação. Este estudo realizou a caracterização ovariana e testicular do peixe Amazônico ornamental Serrapinnus kriegi. Um total de três machos e três fêmeas tiveram suas gônadas analisadas através de microscopia óptica. As fêmeas apresentam ovários preenchidos por oócitos em desenvolvimento assincrônico, indicando desova parcelada da espécie. Além disso, observou-se a formação de micrópila e célula micropilar em oócitos em crescimento primário, representando o desenvolvimento precoce do oócito; a zona radiata nos oócitos vitelogênicos finais é mais espessa em comparação a outras espécies relacionadas, evidenciando o desenvolvimento de uma melhor proteção aos embriões, em função das águas turbulentas que caracterizam seu local de desova nos córregos amazônicos. Os machos apresentam testículos do tipo tubular anastomosado com espermatogônias irrestritas, espalhadas por todo o túbulo seminífero. Os dados apresentados elucidam a dinâmica da espermatogênese e oogênese de uma espécie de peixe ornamental amazônica, por meio da descrição das células germinativas masculinas e femininas.

Oxidative activity of corpus luteum and ovarian parenchyma in Bos taurus indicus heifers

The aim of this study was to evaluate the oxidative stress in ovaries and corpus luteum (CL) of Bos taurus indicus females and the oxidant effect of CL in ovarian tissues in regions near, intermediate, or distant from it. Ovaries (n=12) of Nelore heifers (n=6) were collected from a slaughterhouse and fragmented. Experiment 1, each ovary was obtained from three fragments, resulting in 18 fragments of ovaries with CL (OV+CL) and another 18 fragments of ovaries without CL (OV-CL). Three fragments were generated from CL, totaling 18 CL fragments. In experiment 2, the ovarian fragments were removed from specific regions near, intermediate, or distant from the CL. All the fragments were placed in Eppendorf-type microtubes (1 mL), kept in a thermal container at 4 ºC, and then stored in a -80 ºC freezer for analysis of oxidative stress (TBARS and NBT) and antioxidant potential (FRAP and ABTS). In the antioxidant activity analysis, luteal tissues showed more antioxidant activity than ovarian tissue (FRAP = P < 0.0001; ABTS = P < 0.02). In the oxidative stress analysis, CL had lower levels of reactive oxygen species (ROS; TBARS = P < 0.03; NBT = P < 0.0001) than ovarian tissues. There was no difference in antioxidant activity and oxidative stress between the fragments obtained from different regions (OV+CL versus OV-CL; P > 0.05). The presence of CL in the ovaries of Bos taurus indicus females did not influence the oxidative stress or antioxidant potential of the gonad. Thus, the removal of ovarian fragments with or without the presence of CL indicates that biotechnologies such as in vitro follicle cultivation is possible.(AU)

Carotenoids and retinoids in the gonad of broodstock pikeperch: accumulation during vitellogenesis and influence on egg quality in farmed pikeperch Sander lucioperca

Carotenoids are determinants of reproductive fitness and egg quality. Here we studied the accumulation of astaxanthin (AX), canthaxanthin (CA) zeaxanthin (ZX), lutein (LU), retinol (RX) and dehydroretinol (DR) during vitellogenesis comparing previtellogenic and vitellogenic pikeperch (Sander lucioperca) eggs (n = 5 each), as well as selected tissues (liver, fat and muscles) in first süawning females (1176-1450 g). Futhermore, we compared egg batches with high (88­99% hatching rate, n = 5) or low (40­67% hatching rate, n= 5) egg quality. Vitellogenic follicles revealed higher concentrations of DR, RX, ZX and LU compared to previtellogenic follicles. Neither CA nor AX was detectable. In parallel, DR and RX were mobilized in the liver. In adipose and muscle tissue, comparing previtellogenic and vitellogenic females, no significant differences in carotenoid/retinoid content were observed. In high quality egg batches, both DR and RX were increased. LU was lower in high quality than in low quality eggs. In a conclusion, the amount of retinoids seems suboptimal in low quality egg batches and increased DR and RX are desirable in pikeperch. Since hypervitaminosis of retinoids can be problematic though, supplementation of the food with carotenoids, which can serve as precursors for retinoids, has to be carried out carefully.(AU)

Thecoma in an elderly crioulo mare

Background: Ovarian enlargement in mares can either be a physiological or pathological condition associated with neoplastic or non-neoplastic causes. Neoplasia is more prominent in elderly than young horses. Granulosa cell tumors are the most common neoplastic cause of ovarian enlargement, and thecoma is the rarest. Thecoma is a benign ovarian stromal neoplasia originating from the sexual cords of the embryonic gonad and can affect the production of steroid hormones, leading to changes in behavior and the estrous cycle. If only one ovary is affected, after unilateral ovariectomy, usually the mare can still be used for reproductive purposes. Herein, we report a case of ovarian thecoma in an elderly Crioulo mare. Case: A 15-year-old Crioulo mare was evaluated due to progressive weight loss. The mare presented a low body score, and on physical examination, all parameters were within reference limits. Hematological examination revealed anemia, hyperfibrinogenemia, and neutrophilia. Serum biochemistry examination were within the physiological limits. Rectal palpation revealed an increase in volume in the left ovary (4.3 × 2.9 × 3 cm), smooth consistency and no associated pain in either of the ovaries. Rectal ultrasonography revealed a heterogeneous pattern, suggesting a neoplasm. As a treatment modality and for definitive diagnosis, the mare underwent a unilateral flank ovariectomy laparoscopically in the quadrupedal position. On histopathological evaluation of the left ovary, the sample showed cysts associated with proliferating spindle cells arranged in a solid arrangement, occasionally forming eddies, in addition to hemosiderosis, with clear cytoplasm and lipid vacuoles. Macroscopically, the ovary presented with a regular surface, smooth to firm consistency, a homogeneous brownish appearance with blackened foci, and brownish fluid-filled cysts. These findings confirmed thecoma of the left ovary in the mare. Discussion: Ovarian enlargement in mares can be associated to the presence of neoplasia. Clinical signs of ovarian neoplasms can be nonspecific; therefore, to achieve a definitive diagnosis, it is imperative to evaluate and compare the patient's history, physical examination, and results of complementary examinations. Excessive production of hormones lead to behavioral changes in mares with ovarian tumors, such as masculinized or aggressive behavior, and anestrus persistence. Thecomas may or may not interfere with steroid production and levels. On ultrasound examination, ovarian neoplasms may present similar echogenicity patterns. Histopathological evaluation is the best diagnostic approach after an ovariectomy. The prognosis of ovariectomy is considerably successful with the mare regaining reproductive health, although some mares may become infertile, with a small, inactive contralateral ovary. Thecoma is a rare neoplasm in horses; however, owing to its potential to cause altered reproductive functions, it should be considered as a differential diagnosis in mares with increased ovarian volume.

Intra-abdominal torsion of a non-neoplastic cryptorchid testis in an adult dog / Torção intra-abdominal de testículo criptorquideo sem neoplasia em cão adulto

Intra-abdominal or intrascrotal testicular torsion in dogs occurs due to spermatic cord rotation. Dogs with testicular torsion commonly present severe pain and require surgical intervention. Torsion of intra-abdominal retained testicles in cryptorchid adult dogs is often associated with the presence of testicular neoplasia. Herein, we reported the case of a 5-year-old male poodle with uncommon intra-abdominal testicular torsion (ITT) of a non-neoplastic testicle. The dog was referred to the veterinary hospital with acute abdominal pain in the hypogastric region. An intra-abdominal gonad and alterations compatible with testicular torsion were visualized during ultrasound examination. Orchiectomy and histopathological analysis of the testes confirmed the diagnosis of ITT in the absence of neoplasia. The patient recovered uneventfully from the situation. This report showed that ITT can occur in adult dogs in the absence of testicular neoplasia and reinforce the message that it should be included as a differential diagnosis in cases of acute abdominal pain in cryptorchid dogs. Furthermore, the ultrasound examination in this case of pain in the hypogastric region was decisive for the diagnosis of ITT.

A torção testicular intra-abdominal ou intra-escrotal em cães ocorre devido à rotação do cordão espermático. Os cães com torção testicular apresentam comumente dor intensa e necessitam de intervenção cirúrgica. Em cães adultos, a torção de testículos retidos no abdômen está geralmente associada à presença de neoplasia testicular. Este relato descreve um caso incomum de cão macho, de cinco anos de idade, da raça Poodle, encaminhado ao Hospital Veterinário com dor abdominal aguda na região hipogástrica. Durante o exame ecográfico, foi possível evidenciar um testículo e alterações compatíveis com torção testicular. A orquiectomia intra-abdominal e posterior análise histopatológica confirmaram a ITT sem neoplasia associada. O paciente apresentou evolução clínica satisfatória. Este relato mostra que a ITT na ausência de neoplasia pode ocorrer em cães adultos, reforçando a mensagem de que deve ser considerada como um diferencial diagnóstico em casos de dor abdominal aguda. Além disso, o exame ultrassonográfico realizado neste caso de dor na região hipogástrica foi decisivo para o diagnóstico de ITT.

Desenvolvimento tecnológico no cultivo de filosomas de lagostas palinurídeos / Technological development in the culture of phyllosoma of palinurid spiny lobster / Desarrollo tecnológico en lo cultivo de filosomas de langostas palinurídeos

O desenvolvimento larval completo dos estágios da filosoma da lagosta palinurídeo foi feito em condições de laboratório por vários pesquisadores. A duração dos estágios larvais parece ser específica em relação às espécies. Para as larvas filosomas recentes podem ser fornecidas náuplios de Artemia em combinação com pedaços de gônadas de mexilhões (Mytilus edulis). Gônadas de mexilhões podem ser utilizados exclusivamente após o segundo ou terceiro instar. Grande mortalidade pode ser observada no primeiro instar. A par disso, poucas espécies foram cultivadas devido ao seu dilatado e complexo desenvolvimento larval. A proposta desta revisão é resumir as informações sobre os estudos do cultivo de filosomas.(AU)

The complete larval development of the phylosoma stages of palinurid spiny lobster has been made under laboratory condition by several researchers. Duration of the larval stage seems to be species specific. For the early phyllosomas, food can be consisted of Artemia nauplii in combination with pieces of mussel (Mytilus edulis) gonad. Mussel gonad can be used exclusively after the 2nd or 3rd instar. Heavy mortalities can be noted at the 1st instar. Few spiny lobsters have been raised from egg to puerulus in the lab. Spiny lobsters have a long and complex larval development. The purpose of this review is to sumarize the studies on phyllosoma culture.(AU)

El desarrollo larvario completo de los estadios de filosoma de la langosta espinosa palinúrida ha sido realizado en condiciones de laboratorio por varios investigadores. A duración de las etapas larvais parece ser específica en relacion a las especies. Para las larvas filosomas recientes pueden ser ofrecidas nauplios de Artemia en cobinacion con pedazos de gónadas de mejillones (Mytilus edulis). Gónadas de mejillones pueden ser utilizados exclusivamente após o segundo el tercero instar. Grande mortalidad puede ser observada en lo primeiro instar. Al mismo tiempo, pocas especies han sido cultivadas debido a su dilatado y complejo desarrollo larval. La propuesta desta revisión es resumir la información sobre los estúdios sobre el cultivo de filosomas.(AU)

Reproductive alterations of Biomphalaria glabrata (Say, 1818) infected with Angiostrongylus cantonensis (Chen, 1935) and exposed to Euphorbia milii var. hislopii latex / Alterações na atividade reprodutiva de Biomphalaria glabrata (Say, 1818) infectada com Angiostrongylus cantonensis (Chen, 1935) e exposta ao látex de Euphorbia milii var. hislopii

The natural phytochemical latex of Euphorbia milii var. hislopii is one of the most promising natural molluscicides for the control of Biomphalaria glabrata, and has been widely studied under laboratory conditions for selective control of schistosomiasis transmission. However, the effect of this product on B. glabrata infected by other helminths had not yet been investigated. The present study reports evaluation of the effect of E. milii var. hislopii latex on the survival and reproductive activity of B. glabrata infected by Angiostrongylus cantonensis. For this purpose, the following groups were formed: control (C), exposed (E), infected (I) and infected and exposed for different time intervals (1 day - I+E-1D, 7 days - I+E-7D, 14 days - I+E-14D, 21 days - I+E-21D and 28 days - I+E-28D). The experimental infection was performed with L1 larvae of A. cantonensis and exposure to 0.08 mg/L (LC50) of E. milii latex for a period of 24 hours. We analyzed the effects of E. milii latex on the survival of snails during four weeks, reproductive parameters and possible histophysiopathological changes in the gonad and albumen gland of the snails. Regarding survival, there was reduction of more than 50% in the groups exposed to latex (E and I + E) compared to the control group. As for the number of ovigerous masses, eggs, and average number of hatched snails, significant increases were observed in the I+E-1D group at the 4th week compared to the control group and the other weeks in the same group. Angiostrongylus cantonensis larvae were observed in the gonad and albumen gland from day 21 and 28 of infection in groups I and I+E, respectively, with granuloma-like formation. At these observation periods and in these groups, an increase in galactogen was observed in the albumen gland, which influenced egg laying, suggesting the existence of a fecundity compensation mechanism phenomenon. It was possible to conclude that both stressors - A. cantonensis infection and exposure to E. milii latex - directly influenced the survival and reproductive parameters of B. glabrata.

O fitoquímico natural látex de Euphorbia milii var. hislopii é um dos mais promissores moluscicidas naturais para o controle de Biomphalaria glabrata, tendo sido amplamente estudado em condições laboratoriais como controle seletivo da transmissão da esquistossomose. No entanto, o efeito deste produto em B. glabrata infectada por outro helminto, ainda não tinha sido investigado. O presente trabalho tem por objetivo avaliar o efeito do látex de E. milii var. hislopii na sobrevivência e atividade reprodutiva de B. glabrata infectada por Angiostrongylus cantonensis. Para isso, foram formados os seguintes grupos: controle (C), exposto (E), infectado (I) e infectado e exposto em diferentes intervalos de tempo de infecção (1 dia - I+E-1D, 7 dias - I+E-7D, 14 dias - I+E-14D, 21 dias - I+E-21D e 28 dias - I+E-28D). A infecção experimental foi realizada com larvas L1 de A. cantonensis e a exposição com 0,08 mg/L (LC50) do látex de E. milii por um período de 24 horas. Foram analisados os efeitos do látex de E. milii quanto à sobrevivência dos moluscos ao longo de quatro semanas, aos parâmetros reprodutivos e as possíveis alterações histofisiopatológicas na gônada e na glândula de albúmen dos moluscos analisados. Em relação à sobrevivência foi possível observar redução de mais de 50% nos grupos expostos ao látex (E e I + E) em comparação ao grupo controle. Quanto ao número de massas ovígeras, ovos e número médio de moluscos eclodidos pode-se observar um aumento significativo no grupo I+E-1D na 4ª semana em comparação ao grupo controle e as demais semanas no mesmo grupo. Larvas de A. cantonensis foram observadas na glândula e na glândula de albúmen, a partir dos 21º dias e 28º dia de infecção nos grupos I e I+E, respectivamente, com a formação de granuloma-like. Nesses períodos de observação e nesses grupos pode-se observar na glândula de albúmen, um aumento do conteúdo de galactogênio, o que influenciou a postura dos ovos, sugerindo um fenômeno de compensação da fecundidade. Foi possível concluir que ambos agentes estressores, infecção por A. cantonensis e exposição ao látex de E. milii influenciou diretamente a sobrevivência e os parâmetros reprodutivos de B. glabrata.

Biobanking and use of gonadal tissues - a promising strategy for conserving wildlife from the Caatinga biome

Biological Resource Banks (BRB) or Genetic Resource Banks (GRB) are critical tools for the conservation of animal biodiversity. According to the International Union for Conservation of Nature, more than 38,500 species are threatened with extinction, out of a total of 138,300 surveyed species. These banks are repositories of biological samples and data recovered and preserved for the long term by zoos, universities, research centers and other conservation organizations. In recent years, BRB have increasingly included ovarian and testicular tissues as additional options to rescue and propagate wild species, especially those at risk of extinction. After in vitro culture or grafting, gonadal tissues are potential sources of matured gametes that can be used for Assisted Reproduction Technologies while informing about gametogenesis or mechanisms involved in infertility. It therefore is crucial to properly recover, cryopreserve, and culture these tissues using species-specific protocols. Developing BRBs is currently one of the strategies to preserve species from the Caatinga biome - an exclusively Brazilian biome with a rich wild fauna that suffers from anthropogenic activities. Among wild species from this biome, studies have been primarily conducted in collared peccaries, agoutis, cavies, and armadillos to preserve their ovarian and testicular tissues. Additionally, domestic species such as the domestic cat and donkeys have been proposed as models for wild species that are phylogenetically close. This review addresses the main technical aspects involved in obtaining BRB derived from gonadal tissues in some wild species of the Caatinga biome. It reports recent advances and perspectives to use these biological materials for wildlife conservation.(AU)

Morphology and immunolocalization of intertubular steroidogenic cell in mesonephros of Podocnemis expansa during gonadal differentiation

Sex steroid hormones are critical in gonadal differentiation in turtles. The gonads are not the only organs responsible for producing these hormones during this phase. Mesonephros play an important role in steroidogenesis. The present study aimed to investigate the presence of steroidogenic cells in mesonephros of Podocnemis expansa during gonadal differentiation and to evaluate their morphology and ultrastructure. Ten embryos of P. expansa were collected from 5 nests on day 36 of incubation, during spawning period on an artificial beach. Embryos were extracted from eggs by slicing the shell and euthanized. They were dissected under a stereoscopic microscope to collect the gonad-mesonephro complex, in which were fixed and subsequently processed for light microscopy, immunohistochemistry and transmission electron microscopy analysis. During histological analysis was observed mesonephros has typical morphological structure. Immunohistochemistry showed immunoreaction to aromatase in cells of intertubular space. Confirming these findings, it was possible to observe a type of intertubular cell in several regions of mesonephro, being more predominant in region close to blood vessels, distal and proximal tubules. In ultrastructural analysis these cells were characterized by having a clear, large, and rounded nucleus with evident nucleolus and cytoplasm rich in electron-dense droplets. This study demonstrated for the first time the presence of cells with morphological, immunohistochemical and ultrastructural characteristics similar to steroid-producing cells in P. expansa mesonephrons, suggesting that this organ may contribute to gonadal differentiation in this species.(AU)

Ovarian development of Xiphopenaeus kroyeri (Crustacea: Penaeidae) from Espírito Santo, southeastern Brazil / Desenvolvimento ovariano de Xiphopenaeus kroyeri (Crustacea: Penaeidae) no Espírito Santo, sudeste do Brasil

This study aimed to describe and characterize the stages of gonadal development of females of Xiphopenaeus kroyeri caught by artisanal fishers in Espírito Santo state, southeastern region of Brazil. All females (n= 1,831) were subjected to macroscopic and microscopic morphological analysis (n= 333) of the ovaries. From the morphology, coloration and degree of turgidity of the fresh ovary, macroscopic analysis determined five stages of gonadal development. The macroscopic analysis showed difficulties in differentiating the immature and spawning stages due to the similarity between the colors of the ovaries, which confirms the need to perform the macroscopic and histological analysis simultaneously for fisheries management studies. Microscopic observations allowed us to analyze the following six stages of cell development: oogonia, previtellogenic oocytes, primary vitellogenic oocytes, secondary vitellogenic oocytes, mature oocytes and atretic oocytes. From this, five stages of gonadal development were defined, i.e., immature, early development, advanced development, mature and spawned. The presence of peripheral bodies was not observed in this species. These results help to clarify and better understand the reproductive and population aspects of the Atlantic Seabob, which are fundamental for the establishment of management and conservation measures of this resource.

Este estudo teve como objetivo descrever e caracterizar os estágios de desenvolvimento gonadal de fêmeas de Xiphopenaeus kroyeri capturado por pescadores artesanais no Estado do Espírito Santo, região sudeste (Região sudeste = Rio de Janeiro, São Paulo, Minas Gerais e Espírito Santo) do Brasil. Todas as fêmeas (n= 1.831) foram submetidas à análise morfológica macroscópica e microscópica (n= 333) dos ovários. A partir da morfologia, coloração e grau de turgidez do ovário fresco, a análise macroscópica determinou cinco estágios de desenvolvimento gonadal. A análise macroscópica mostrou dificuldades em diferenciar os estágios imaturo e reprodutivo devido à semelhança entre as cores dos ovários, o que confirma a necessidade de realizar a análise macroscópica e histológica simultaneamente para estudos de manejo pesqueiro. As observações microscópicas permitiram analisar os seguintes seis estágios de desenvolvimento celular: oogônias, ovócitos prévitelogênicos, ovócitos em vitelogênese primária, ovócitos em vitelogênese secundária, ovócitos maduros e ovócitos atrésicos. A partir disso, foram definidos cinco estágios de desenvolvimento gonadal, ou seja, imaturo, desenvolvimento inicial, desenvolvimento avançado, maduro e desovado. A presença de corpos periféricos não foi observada nesta espécie. Estes resultados ajudam a esclarecer e compreender melhor os aspetos reprodutivos e populacionais do camarão do Atlântico, que são fundamentais para o estabelecimento de medidas de gestão e conservação deste recurso.

Sexual maturity of Callinectes danae and C. ornatusfrom Paraguaçu river estuary, Bahia state, Brazil / Maturidade sexual de Callinectes danaee C. ornatusno estuário do rio Paraguaçu, Bahia, Brasil

Callinectes danae and C. ornatus are economically important species in several countries, including Brazil. Therefore, this study aimed to estimate the L50 of these species using morphological, morphometric, and physiological data. The estimated values were statically compared and results from previous studies were analyzed to verify latitudinal variations among the sizes of sexual maturity at different latitudes of the Brazilian coast. Individuals were monthly collected in the Paraguaçu River estuary from August 2013 to July 2014, using cage traps. L50 estimates for C. danae were morphological=55.80 mm, morphometric=59.04, mm, and physiological=60.41 mm for males and morphological=54.63 mm, morphometric=55.33 mm, and physiological=57.29 mm for females. Considering C. ornatus, estimates were morphological=42.63 mm, morphometric=50.81 mm, and physiological=43.95 mm for males and morphological=42.33 mm, mor-phometric=42.75 mm, and physiological=40.43 mm for females. Our results indicated that the minimum catch sizes should be equal to 61.00 mm for C. danae and 51.00 for C. ornatus and that the mesh of traps used by fishermen must be adjusted to prevent the capture of immature animals.

Callinectes danae e C. ornatus são siris economicamente importantes em vários países, incluindo o Brasil. O presente estudo objetivou determinar o L50 dessas espécies com base em dados morfológicos, morfométricos e fisiológicos. Os valores estimados foram comparados estatisticamente e os resultados de estudos anteriores no Brasil foram analisados para verificar variações latitudinais nos tamanhos de maturidade sexual em diferentes regiões da costa brasileira. Os exemplares foram coletados mensalmente, de agosto de 2013 a julho de 2014, por meio de armadilhas do tipo gaiola. As estimativas de L50 em C. danae foram morfológica=55,80 mm, morfométrica=59,04 mm e fisiológica=60,41 mm para machos e morfológica=54,63 mm, morfométrica=55,33 mm e fisiológica=57,29 mm para fêmeas. Considerando C. ornatus, as estimativas foram: morfológica=42,63 mm, morfométrica=50,81 mm e fisiológica=43,95 mm para machos e morfológica=42,33 mm, morfométrica=42,75 mm e fisiológica=40,43 mm para fêmeas. Nossos resultados indicaram que os tamanhos mínimos de captura deveriam ser iguais a 61,00 mm para C. danae e 51,00 mm para C. ornatus e a malha das armadilhas usadas pelos pescadores ajustada para impedir a captura de animais imaturos.

Aquatic pollutants are associated with reproductive alterations and genotoxicity in estuarine fish (Sciades herzbergii - Bloch, 1794) from the Amazon Equatorial Coast / Poluentes aquáticos estão associados a alterações reprodutivas e genotoxicidade em peixe estuarino (Sciades herzbergii - Bloch, 1794) da Costa Equatorial Amazônica

Estuaries are constantly subject to continuous environmental impacts of human activities, such as fisheries, port or industry, and domestic sewage, with fish being one of the most affected aquatic animals, reflecting the impacts directly on their bodies. Thus, the aim of this study was to carry out the biomonitoring of an estuary located on the Amazonian Equatorial Coast through analysis of PAHs (Polycyclic Aromatic Hydrocarbons) in the water, in addition to trace metals, histopathological alterations and analysis of erythrocyte micronuclei in Sciades herzbergii. S. herzbergii was used as a model species, due to its estuarine-resident behavior. Gonad and gill samples were subjected to histopathological evaluations. The quantification of trace metals was performed in samples of skeletal muscles of the animals collected, where concentrations of Lead (Pb), Copper (Cu), Zinc (Zn), Cadmium (Cd), Magnesium (Mg), Iron (Fe) and Aluminum (Al) were found. Except for Cadmium (Cd), all the concentrations were above the recommended limits. The PAHs analysis revealed the presence of Naphthalene and Acenaphthene in the water samples Histopathological and genotoxic analyses revealed of lesions in 100% of the study specimens. Thus, the histological and genotoxic alterations found in 100% of S. herzbergii specimens captured in São José Bay-MA are potentially associated with PAH concentrations present in the water. These results are potentially associated with the presence of PAH and trace metals, both in water and in animal tissues, inferring a general scenario of environmental contamination which directly implies a risk to the health and survival of the local biota. This study shows the relevance of continuous biomonitoring of estuarine ecosystems, in order to guide authorities regarding sewage management and ensure the evolutionary development of estuarine species, especially fishes of importance in the local cuisine, therefore related to human food security.

Os estuários estão constantemente sujeitos a impactos ambientais de atividades antropogênicas, como pesca, movimentação portuária, indústria ou esgoto doméstico, onde os peixes são um dos animais aquáticos mais atingidos, sendo capazes de refletir os impactos diretamente em seu organismo. Assim, o objetivo desse estudo foi realizar o biomonitoramento de um estuário da Costa Equatorial Amazônica por meio de análises de PAHs (Hidrocarboneto Policiclico Aromáticos) na água, além de metais traços, alterações histopatológicas e análises de micronúcleos eritrocitários em Sciades herzbergii.. O S. herzbergii foi utilizado como espécie modelo, devido seu comportamento estuarino residente. Amostras de gônadas e brânquias foram submetidas a avaliações histopatológicas. A quantificação de metais traços foi realizada em amostras de músculos esqueléticos dos animais coletados, onde foram encontrados teores de Chumbo (Pb), Cobre (Cu), Zinco (Zn), Cádmio (Cd), Magnésio (Mg), Ferro (Fe) e Alumínio (AL), e foram encontradas em concentrações acima dos limites recomendados, à exceção do Cádmio (Cd). A análise dos PAHs revelou a presença de Naftaleno e Acenafteno nas amostras de água. Análises histopatológicas e genotóxicas revelaram lesões em 100% dos espécimes estudados, resultados estes, potencialmente associados à presença de PAH e de metais traços, tanto na água quanto nos tecidos animais, comprovando um cenário geral de contaminação ambiental implicando diretamente em risco à saúde e sobrevivência da biota local. Esse estudo mostra a relevância do biomonitoramento contínuo dos ecossistemas estuarinos, afim de nortear autoridades no manejo de efluentes e assegurar o desenvolvimento evolutivo de espécies estuarinas, sobretudo dos peixes de importância na culinária regional, diretamente relacionados à segurança alimentar humana.

Potential use of bacterial pigments as anticancer drugs and female reproductive toxicity: a review / Uso potencial de pigmentos bacterianos como drogas anticâncer e toxicidade reprodutiva feminina: uma revisão

Natural bioactive compounds obtained from microorganisms, have awakened particular interest in the industry nowadays. This attention comes when natural resources depletion is pronounced, and the acquisition of both new plant origin resources and bioactive products, represents a challenge for the next generations. In this sense, prospecting for large-scale production and use of bacterial pigments is a necessary strategy for the development of novel products. A wide variety of properties have been attributed to these substances and, among them, their therapeutic potential against important diseases, such as cancer. There is consensus that available chemotherapy protocols are known to detrimentally affect cancer patients fertility. Hence, considerable part of the deleterious effects of chemotherapy is related to the drugs cytotoxicity, which, in addition to cancer cells, also affect normal cells. Therefore, the intrinsic properties of bacterial pigments associated with low cytotoxicity and relevant cell selectivity, certified them as potential anticancer drugs. However, little information is available about reproductive toxicity of these new and promising compounds. Thus, the present review aims to address the main bacterial pigments, their potential uses as anticancer drugs and their possible toxic effects, especially on the female gonad.

Os compostos bioativos naturais obtidos de microrganismos têm despertado especial interesse da indústria nos últimos anos. Esta atenção ocorre em um momento em que o esgotamento de recursos naturais é pronunciado, e a aquisição de novos insumos e produtos bioativos de origem vegetal representa um desafio para as próximas gerações. Neste sentido, a prospecção para a produção e uso em larga escala dos pigmentos bacterianos tem representado uma importante estratégia para o desenvolvimento de novos produtos. Uma grande variedade de propriedades foi atribuída a estas substâncias, entre elas, o potencial terapêutico contra doenças importantes, como o câncer. Existe um consenso de que os protocolos quimioterápicos disponíveis são conhecidos por afetarem negativamente a fertilidade de pacientes com câncer. Grande parte dos efeitos deletérios da quimioterapia está relacionado à citotoxicidade das drogas usadas para este fim, que além das células cancerosas, afetam as células normais. Nesse sentido, as propriedades naturais atribuídas aos pigmentos bacterianos associadas à baixa citotoxicidade e relevante seletividade, os qualificaram como potenciais drogas anticâncer. No entanto, pouco se tem de informação a respeito da toxicidade reprodutiva destes novos e promissores compostos. Dessa forma, a presente revisão tem o objetivo de abordar os principais pigmentos bacterianos, suas utilizações potenciais como drogas anticâncer, bem como os seus possíveis efeitos tóxicos, sobretudo, sobre a gônada feminina.

Gonadal development and sexuality of Larkinia grandis (Arcida: Arcidae) inhabiting southeastern Gulf of California

ABSTRACT Larkinia grandis (Broderip & G.B. Sowerby I, 1829), an important fishing resource for Mexican communities, is an Arcidae clam. It is also considered a species with aquaculture potential. In this work we investigated the gonadal phases and sexuality in a population of L. grandis in the Gulf of California. Our findings support the hypothesis that there is one male per female in the population studied. It also documents that the shape, position and color of the gonads of L. grandis are consistent with observations in other Arcidae species. Additionally, five gonadal phases are differentiated and described in males and females (development, mature, spawning, post-spawning and resting), with a noticeable presence of brown cells during post-spawning and the onset of the resting phase, suggesting that those cells are involved in the reabsorption of remnants. Additionally, asynchronous gametogenesis in males, synchronic gametogenesis in females and batch spawning are defined. The results of this contribution can be used in the efforts to protect this bivalve.

Testisacs microanatomy and spermatogenesis of Helobdella simplex (Hirudinida: Glossiphoniidae)

The aim of this study was to describe the testisac microanatomy and spermatogenesis of Helobdella simplex (Moore, 1911). The leeches were collected in the Sauce Chico River (Tornquist) during April 2012. The collection of material was carried out manually by checking rocks, logs, leaves and artificial substrates in the river bed. Helobdella simplex has six pairs of testisacs located between the gastric caeca. The testisac wall has two structurally and functionally differentiated cell types: the lining mesothelial cells and the germline cells. The lining cells are flat and form a continuous layer. In H. simplex no ciliated cells were observed into the testisacs, nor the formation of spermatogonia due to detachment of the testicular wall. Cell types were microscopically characterized, and five spermatogenesis stages are described, taking into account the maturation of germinal cells, the morphological changes in the cytophore and phagocytic activity. Testicular phagocytes actively participate during spermatogenesis. They are involved both in the removal of defective cells during the early stages of spermatogenesis and in the removal of the cytophore during the reacting reabsorption. Our study adds the knowledge of spermatogenesis in H. simplex and might be useful to differentiate the stages of development during the reproductive cycle, and to interpret aspects of its population dynamics.(AU)

Testisacs microanatomy and spermatogenesis of Helobdella simplex (Hirudinida: Glossiphoniidae)

ABSTRACT The aim of this study was to describe the testisac microanatomy and spermatogenesis of Helobdella simplex (Moore, 1911). The leeches were collected in the Sauce Chico River (Tornquist) during April 2012. The collection of material was carried out manually by checking rocks, logs, leaves and artificial substrates in the river bed. Helobdella simplex has six pairs of testisacs located between the gastric caeca. The testisac wall has two structurally and functionally differentiated cell types: the lining mesothelial cells and the germline cells. The lining cells are flat and form a continuous layer. In H. simplex no ciliated cells were observed into the testisacs, nor the formation of spermatogonia due to detachment of the testicular wall. Cell types were microscopically characterized, and five spermatogenesis stages are described, taking into account the maturation of germinal cells, the morphological changes in the cytophore and phagocytic activity. Testicular phagocytes actively participate during spermatogenesis. They are involved both in the removal of defective cells during the early stages of spermatogenesis and in the removal of the cytophore during the reacting reabsorption. Our study adds the knowledge of spermatogenesis in H. simplex and might be useful to differentiate the stages of development during the reproductive cycle, and to interpret aspects of its population dynamics.

The neuroendocrine pathways and mechanisms for the control of the reproduction in female pigs

Abstract Within the hypothalamic-pituitary-gonad (HPG) axis, the major hierarchical component is gonadotropin-releasing hormone (GnRH) neurons, which directly or indirectly receive regulatory inputs from a wide array of regulatory signals and pathways, involving numerous circulating hormones, neuropeptides, and neurotransmitters, and which operate as a final output for the brain control of reproduction. In recent years, there has been an increasing interest in neuropeptides that have the potential to stimulate or inhibit GnRH in the hypothalamus of pigs. Among them, Kisspeptin is a key component in the precise regulation of GnRH neuron secretion activity. Besides, other neuropeptides, including neurokinin B (NKB), neuromedin B (NMB), neuromedin S (NMS), α-melanocyte-stimulating hormone (α-MSH), Phoenixin (PNX), show potential for having a stimulating effect on GnRH neurons. On the contrary, RFamide-related peptide-3 (RFRP-3), endogenous opioid peptides (EOP), neuropeptide Y (NPY), and Galanin (GAL) may play an inhibitory role in the regulation of porcine reproductive nerves and may directly or indirectly regulate GnRH neurons. By combining data from suitable model species and pigs, we aim to provide a comprehensive summary of our current understanding of the neuropeptides acting on GnRH neurons, with a particular focus on their central regulatory pathways and underlying molecular basis.

Desenvolvimento e diferenciação gonadal em embriões de Podocnemis unifilis (Testudines: Podocnemididae) em temperatura controlada / Development and gonadal differentiation in embryos of Podocnemis unifilis (Testudines: Podocnemididae) at controlled temperature

Esse estudo objetivou caracterizar o desenvolvimento e a diferenciação gonadal de Podocnemis unifilis em temperatura controlada de 32°C em incubadora artificial e com controle de umidade feito por um higrômetro interno. Foram realizadas análises macroscópicas, por microscopia de luz e por microscopia eletrônica de transmissão. O desenvolvimento gonadal foi agrupado em três etapas: células germinativas primordiais (CGP), estabelecimento da gônada indiferenciada e diferenciação gonadal. As CGP só foram visualizadas a partir do 7º dia de incubação, e se encontravam na região caudal do corpo do embrião, e entre os dias 9 e 11 de incubação foi possível observar a migração das CGP a partir do saco vitelínico em direção à região ventromedial do mesonefro. Com 20 dias de desenvolvimento, a gônada indiferenciada estava estabelecida, sendo identificadas duas regiões distintas: a região cortical externa, caracterizada pela presença de CGP, e a região medular interna, marcada por cordões sexuais primitivos. Nesse estudo, a diferenciação gonadal só ocorreu em testículo. A diferenciação teve início após 35 dias de incubação, com o início da organização dos cordões sexuais em túbulos seminíferos. As etapas de diferenciação gonadal foram semelhantes às de outras espécies de quelônios com determinação do sexo pela temperatura, no entanto, neste estudo, usando a temperatura pivotal descrita para a espécie, todos os indivíduos analisados diferenciaram-se em machos, sendo necessária, portanto, a realização de mais estudos com diferentes temperaturas.(AU)

This study aimed to characterize the development and gonadal differentiation of Podocnemis unifilis at a controlled temperature of 32° C in artificial incubator and with humidity control performed by an internal hygrometer. Macroscopic analyzes, light microscopy and transmission electron microscopy were performed. We grouped gonadal development into three stages: primordial germ cells (PGC), establishment of the undifferentiated gonad and gonadal differentiation. We only visualized the PGC from the 7th day of incubation, and in the caudal region of the embryo body; between days 9 and 11 of incubation we were able to observe the migration of the PGC from the viteline sac towards the ventromedial region of the mesonefro. At 20 days of development, the undifferentiated gonad was established and two distinct regions were identified: the external cortical region characterized by the presence of PGC, and the internal medullary region marked by primitive sexual cords. In this study, only gonadal differentiation occurred in the testicle. Gonadal differentiation began after 35 days of incubation with the beginning of the organization of sexual cords in the seminiferous tubules. The stages of gonadal differentiation were similar to those of other species of turtles with temperature determined sex, however in this study, using the pivotal temperature described for the species all the individuals analyzed were males, and it is there for necessary to carry out further studies using a higher incubation temperature.(AU)

Desenvolvimento e diferenciação gonadal em embriões de Podocnemis unifilis (Testudines: Podocnemididae) em temperatura controlada / Development and gonadal differentiation in embryos of Podocnemis unifilis (Testudines: Podocnemididae) at controlled temperature

Esse estudo objetivou caracterizar o desenvolvimento e a diferenciação gonadal de Podocnemis unifilis em temperatura controlada de 32°C em incubadora artificial e com controle de umidade feito por um higrômetro interno. Foram realizadas análises macroscópicas, por microscopia de luz e por microscopia eletrônica de transmissão. O desenvolvimento gonadal foi agrupado em três etapas: células germinativas primordiais (CGP), estabelecimento da gônada indiferenciada e diferenciação gonadal. As CGP só foram visualizadas a partir do 7º dia de incubação, e se encontravam na região caudal do corpo do embrião, e entre os dias 9 e 11 de incubação foi possível observar a migração das CGP a partir do saco vitelínico em direção à região ventromedial do mesonefro. Com 20 dias de desenvolvimento, a gônada indiferenciada estava estabelecida, sendo identificadas duas regiões distintas: a região cortical externa, caracterizada pela presença de CGP, e a região medular interna, marcada por cordões sexuais primitivos. Nesse estudo, a diferenciação gonadal só ocorreu em testículo. A diferenciação teve início após 35 dias de incubação, com o início da organização dos cordões sexuais em túbulos seminíferos. As etapas de diferenciação gonadal foram semelhantes às de outras espécies de quelônios com determinação do sexo pela temperatura, no entanto, neste estudo, usando a temperatura pivotal descrita para a espécie, todos os indivíduos analisados diferenciaram-se em machos, sendo necessária, portanto, a realização de mais estudos com diferentes temperaturas.

This study aimed to characterize the development and gonadal differentiation of Podocnemis unifilis at a controlled temperature of 32° C in artificial incubator and with humidity control performed by an internal hygrometer. Macroscopic analyzes, light microscopy and transmission electron microscopy were performed. We grouped gonadal development into three stages: primordial germ cells (PGC), establishment of the undifferentiated gonad and gonadal differentiation. We only visualized the PGC from the 7th day of incubation, and in the caudal region of the embryo body; between days 9 and 11 of incubation we were able to observe the migration of the PGC from the viteline sac towards the ventromedial region of the mesonefro. At 20 days of development, the undifferentiated gonad was established and two distinct regions were identified: the external cortical region characterized by the presence of PGC, and the internal medullary region marked by primitive sexual cords. In this study, only gonadal differentiation occurred in the testicle. Gonadal differentiation began after 35 days of incubation with the beginning of the organization of sexual cords in the seminiferous tubules. The stages of gonadal differentiation were similar to those of other species of turtles with temperature determined sex, however in this study, using the pivotal temperature described for the species all the individuals analyzed were males, and it is there for necessary to carry out further studies using a higher incubation temperature.