RESUMEN: El BPA es un disruptor endocrino multifuncional con presencia ubicua en los ecosistemas acuáticos. La Meseta Central mexicana habitada por peces vivíparos endémicos, es una zona severamente impactada por la contaminación. Sin embargo, los esfuerzos por comprender los efectos del BPA en especies nativas como Goodea atripinnis son inexistentes. Este estudio se centró en proporcionar evidencia in vivo de alteraciones en los testículos de machos de G. atripinnis debido a la exposición aguda al BPA en concentraciones de prueba de 1 mg/L, 10 mg/L y 50 mg/L durante 96 h. La exposición a BPA 1 mg/L y 10 mg/L mostró degeneración y desorganización en el tejido germinal. Además, hubo una disminución notable de los espermatozoides dentro de los túbulos seminíferos de machos expuestos a 10 mg/L de BPA. En todos los tratamientos las células somáticas presentaron alteraciones por engrosamiento del tejido conectivo y aumento de las fibras de colágeno. Además, se produjo inflamación y sangrado en los testículos de machos expuestos a 1 y 10 mg/L de BPA. Las alteraciones en los testículos de G. atripinnis están relacionadas con la toxicidad del BPA, lo que puede provocar apoptosis en las células germinales aumentando el tejido conectivo. Finalmente, si bien los cambios producidos por el BPA se hicieron evidentes en la exposición aguda (96 h), sus efectos probablemente sean irreversibles, comprometiendo la reproducción de G. atripinnis.

SUMMARY: BPA is a multifunctional endocrine disruptor with ubiquitous presence in aquatic ecosystems. The Mexican Central Plateau is an area severely impacted by pollution, inhabited by endemic viviparous fish. However, efforts to understand the effects of BPA on native species such as Goodea atripinnis are non-existent. This study focused on providing in vivo evidence of alterations in the testes of G. atripinnis males due to acute exposure to BPA at test concentrations of 1 mg/L, 10 mg/L, and 50 mg/L for 96 h. BPA exposition 1 mg/L and 10 mg/L showed degeneration and disorganization in germinal tissue. Furthermore, there was a notable decrease in sperm within the seminiferous tubules of males exposed to 10 mg/L of BPA. In all treatments, somatic cells had alterations by connective tissue thickening and an increase in collagen fibers. Additionally, inflammation and bleeding occurred in the testes of males exposed to 1 and 10 mg/L BPA. The alterations in the testes of G. atripinnis are related to BPA toxicity, which can lead to apoptosis in germ cells increasing connective tissue. Finally, even though the changes produced by BPA became evident in acute exposure (96 h), its effects are probably irreversible, compromising the reproduction of G. atripinnis.

Resumo Aspectos da biologia reprodutiva de Donax striatus foram estudados. Os indivíduos foram coletados mensalmente de fevereiro de 2021 a janeiro de 2022 na praia Gado Bravo, no município de Tibau do Norte, estado do Rio Grande do Norte, Brasil. Donax striatus é uma espécie dióica sem dimorfismo macroscópico externo (na concha) ou interno (gônadas). Assim, é necessário um exame microscópico das células reprodutivas. Para a caracterização dos estágios de desenvolvimento gonadal e determinação do tamanho na primeira maturidade sexual (L50), 30 espécimes foram selecionados mensalmente entre fevereiro de 2021 e janeiro de 2022 e submetidos ao processamento histológico. O índice de Condição (IC) de cada indivíduo foi estimado e as variações mensais foram estatisticamente avaliadas. O tamanho de primeira maturação (L50) foi estimado em 14,2 mm de comprimento de concha. Para promover a conservação da espécie, recomenda-se a captura de indivíduos maiores que 14,2 mm. Os menores índices de condição foram encontrados na estação seca, com maior ocorrência de organismos em fase de eliminação e exibindo reorganização do tecido gonadal. Índices maiores foram encontrados no período chuvoso, com presença de indivíduos maduros. A natureza contínua da gametogênese em Donax stritatus reflete a influência das chuvas na região. Machos e fêmeas apresentam pico de eliminação de gametas com pausas durante o ano, mas com presença de indivíduos em maturação e eliminação ao longo do ano. Como a mariscagem visando Donax striatus é excessiva na Praia de Gado Bravo, no estado do Rio Grande do Norte, espera-se que os resultados do presente estudo possam contribuir para o estabelecimento de medidas de manejo da atividade e estratégias de conservação da espécie. estudados 202 Bravo Norte Brasil concha gônadas. gônadas . (gônadas) Assim reprodutivas L50, L50 L , 3 histológico IC (IC avaliadas (L50 142 14 2 14, recomendase recomenda se seca chuvoso maduros região ano esperase espera 20 (gônadas L5 (L5 1 (L

Abstract Aspects of the reproductive biology of Donax striatus were studied from individuals collected from Gado Bravo Beach in the municipality of Tibau do Norte, state of Rio Grande do Norte, Brazil. Donax striatus is a dioic species without external (on the shell) or internal (gonads) macroscopic dimorphism. Thus, a microscopic examination of the reproductive cells is necessary. For the characterization of the gonadal development stages and determination of the size at first sexual maturity (L50), 30 specimens were selected monthly between February 2021 and January 2022 and submitted to histological processing. The condition index (CI) of each individual was estimated and monthly variations were statistically assessed. The size at first maturity (L50) was estimated to be 14.2 mm in shell length. To foster conservation of the species, catches of individuals larger than 14.2 mm is recommended. The lowest condition indices were found in the dry season, with a greater occurrence of organisms in the elimination stage and exhibiting gonad tissue reorganization. Higher indices were found in the rainy season, with the presence of mature individuals. The continuous nature of gametogenesis in Donax stritatus reflects the influence of rainfall in the region. Males and females have peak gamete elimination with pauses during the year, but with the presence of maturing and eliminating individuals throughout the year. As shellfish gathering targeting Donax striatus is excessive on Gado Bravo Beach in the state of Rio Grande do Norte, it is hoped that the results of the present study can contribute to the establishment of management measures for the activity and conservation strategies for the species. Norte Brazil gonads (gonads dimorphism Thus necessary L50, L50 L , 3 202 processing CI (CI assessed (L50 142 14 2 14. length recommended season reorganization region year L5 20 (L5 1 (L

Resumo O polvo com chifres Eledone cirrhosa, uma espécie de tamanho médio encontrada no Golfo Arábico ao largo da Arábia Saudita, foi coletado mensalmente no Golfo Arábico ao largo da cidade de Dammam durante o período de janeiro a dezembro de 2022. As amostras foram dissecadas e preparadas para exame por microscopia eletrônica. Durante a maturação genital, os túbulos seminíferos se estabelecem nos testículos, com espermatogônias ativas se dividindo. Espermatócitos 1 são observados no túbulo, seguidos de aumento de espermatogônias e espermatócitos em agosto. A espermiogênese começa com espermátides esféricas evoluindo para espermátides alongadas. Em setembro, as espermatogônias ativas, as divisões meióticas e o aumento da espermiogênese continuam. Os espermatozoides aparecem na Bolsa de Needham, indicando maturidade sexual. O ovário passa por vários estágios de desenvolvimento, com oócitos no estágio I em junho e julho, seguido pelo estágio II em outubro e novembro. No estágio IIIc, os cordões foliculares invadem o citoplasma do oócito, formando numerosas inclusões lipídicas e grânulos proteicos. O citoplasma contém cisternas de retículo endoplasmático e um Aparelho de Golgi pouco desenvolvido. O estágio IV ocorre em novembro, caracterizado pelo máximo desenvolvimento dos cordões foliculares e pelo início da vitelogênese. O ooplasma contém numerosas inclusões lipídicas, um sincício e células secretoras. A partir de dezembro, os oócitos do estágio V estão principalmente presentes, indicando a fase de atividade de secreção máxima. As plaquetas da gema acumulam-se no ooplasma do ovócito e o córion se forma na zona pelúcida. Em janeiro, os primeiros óvulos lisos são encontrados no ovário de alguns polvos e sua proporção aumenta continuamente. Este estudo teve como objetivo investigar a ação mitogênica da gonadotrofina bem como identificar os períodos de intensa multiplicação celular durante o ciclo sexual por meio de métodos citológicos. cirrhosa Saudita 2022 eletrônica genital testículos dividindo túbulo agosto alongadas setembro continuam Needham julho novembro IIIc oócito proteicos desenvolvido vitelogênese secretoras presentes máxima acumulamse acumulam pelúcida continuamente citológicos 202 20 2

Abstract The horned octopod Eledone cirrhosa, a medium-sized species found in Arabian Gulf off Saudi Arabia, was collected monthly from the Arabian Gulf off Dammam city during January to December 2022. Samples were dissected and prepared for examination using transmission electron microscopy. During genital maturation, seminiferous tubules are established in the testis, with active spermatogonia dividing. Spermatocytes 1 are observed in the tubule, followed by an increase in spermatogonia and spermatocytes in August. Spermiogenesis begins, with spherical spermatids evolving into elongated spermatids. In September, active spermatogonia, meiotic divisions, and increased spermiogenesis continue. Spermatozoa appear in Needham's pouch, indicating sexual maturity. The ovary undergoes various stages of development, with oocytes at stage I in June and July, followed by stage II in October and November. In stage III, follicular cords invade the oocyte's cytoplasm, forming numerous lipid inclusions and protein granules. The cytoplasm contains cisternae of endoplasmic reticulum and a poorly developed Golgi apparatus. Stage IV occurs in November, characterized by the maximum development of follicular cords and the beginning of vitellogenesis. The ooplasm contains numerous lipid inclusions, a syncytium, and secretory cells. From December, stage V oocytes are mainly present, indicating the activity phase of maximum secretion. Yolk platelets accumulate in the oocyte ooplasm, and chorion forms at the zona pellucida. In January, the first smooth eggs are found in some octopuses' ovary, with their proportion increasing steadily. This study aimed to investigate the mitogenic action of gonadotropin and identify the periods of intense cell multiplication during the sexual cycle using cytological methods. cirrhosa mediumsized medium sized Arabia 2022 microscopy maturation testis dividing tubule August begins September divisions continue Needhams Needham s pouch maturity July November III granules apparatus vitellogenesis syncytium cells present secretion pellucida octopuses steadily methods 202 20 2

Abstract Reproductive control is one of the biggest challenges in tilapia production and triploidy was developed as an alternative to sterilization. In general, polyploids present chromosomal instability but for triploid Nile tilapia it has yet to be reported. This study evaluated the chromosomal instability from juveniles to adulthood, growth performance and gonadal status of tilapia hatched from eggs submitted or not to heat shock for triploid induction. Nile tilapia oocytes were fertilized (1,476 oocytes), half of the eggs were subjected to a four-minute shock in 41 °C water four minutes after fertilization and the other half were not (Control group). The eggs were incubated (at 27°C) and 160 larvae from the treated group hatched and survived after yolk sac absorption. The determination of ploidy was performed by flow cytometry at 85th (juveniles) and 301st (adults) days of age post yolk sac absorption. At the time of the first cytometry analysis there were 73 surviving juveniles from the treated group, and only 14 were confirmed triploid. However, at the analysis of adult ploidy, one out of 8 surviving adult tilapias from the 14 confirmed triploid juveniles remained triploid. Gonadal histology showed that the non-remaining triploids continued to produce gametes. The growth performance of triploid tilapia was initially superior to that of diploid tilapia during the juvenile phase, but similar in adults. Once the chromosome sets are lost and the tilapias become diploid again, at least in tissues with a high proliferation rate, such as the hematopoietic tissue that was analyzed (and possibly in gonads), all possible advantages of triploids are probably lost. Thus, our results suggest that, due to genomic instabilities, the triploid generation of tilapia has low efficiency. sterilization general reported adulthood induction 1,476 1476 1 476 (1,47 oocytes, , oocytes) fourminute minute 4 C Control group. . group) 27°C 27C 27 16 absorption th (juveniles st adults (adults 7 However nonremaining non remaining gametes phase again rate gonads, gonads gonads) Thus instabilities efficiency 1,47 147 47 (1,4 2 1,4 (1, 1, (1 (

Abstract The reproductive cycle is an essential aspect of oyster farming. Understanding the period of each sexual stage aids in planning oyster harvest, seed collection with artificial collectors, and reproductive season for hatchery seed production. In this sense, this work evaluated the reproductive cycle of Crassostrea gasar oysters cultivated in three important oyster farming locations in the Estuarine Complex of Paranaguá, Rasa Island, Medeiros, and Ponta Oeste (Mel Island), with monthly oyster sampling in each sampling site. Temperature and salinity were recorded in loco at each sampling time. In the laboratory, oysters were identified and measured; tissue samples were collected for molecular identification of species; and histology was conducted for reproductive cycle analysis. Histological examination of the gonadal tissue was performed with slides stained with Harris hematoxylin and eosin. Salinity and temperature data showed no differences between the three sampling sites during the studied period. Molecular analysis showed that the oysters sampled were C. gasar (100%). Histological analysis showed intense spawning of C. gasar in December and February (26.72 °C) and June (20 °C). There was a resting stage in both males and females during winter. Some oysters (n = 21) were parasitized by Bucephalus sp., and it was impossible to determine oyster sex. harvest collectors production sense Paranaguá Island Medeiros Mel , Island) site time laboratory measured species eosin C 100%. 100 100% . (100%) 26.72 2672 26 72 (26.7 °C 20 (2 °C. winter n 21 sp sp. sex 10 (100% 26.7 267 2 7 (26. ( 1 (100 26. (26 (10 (1

Abstract Clinical case description: This report presents the case of a 32-year-old man with palpable solid nodule in the left testicle with a diagnostic hypothesis of testicular cancer, which turned out to be splenic tissue through pathological anatomy analysis. Relevance: The splenogonadal fusion is a rare and benign congenital anomaly related to the presence of ectopic spleen tissue in the gonadal parenchyma, caused by fusion between these two tissues during the fifth or sixth week of embryonic development. This condition is of great importance because its diagnosis is difficult and often confused with other gonadal disorders such as testicular neoplasm. Clinical implications: The most common consequence of this testicular disorder is the unnecessary orchiectomy, in which a variety of cases are submitted. To avoid testicle removal, intraoperative pathological examination can be used in these cases. Conclusions: The ectopic spleen in testicular parenchyma is a rare condition and its diagnosis is difficult. It is important to consider this condition as a differential diagnosis in testicular disorders, especially in the case of diagnostic hypothesis of testicular cancer, so that testicle removal can be prevented by the use of intraoperative pathological exam.

Resumen Descripción del caso clínico: En este reporte se presenta el caso de un hombre de 32 años con nódulo sólido palpable en testículo izquierdo con hipótesis diagnóstica de cáncer de testículo, que en realidad la anatomía patológica descubrió que se trataba de tejido esplénico. Relevancia: La fusión esplenogonadal es una anomalía congénita rara y benigna relacionada con la presencia de tejido esplénico ectópico en el parénquima gonadal, causada por la fusión entre estos dos tejidos durante la quinta o sexta semana de desarrollo embrionario. Esta condición es de gran importancia porque su diagnóstico es difícil y muchas veces se confunde con otros trastornos gonadales como la neoplasia testicular. Implicaciones clínicas: La consecuencia más frecuente de este trastorno testicular es la orquiectomía innecesaria, a la que se someten diversos casos. Para evitar la extirpación del testículo, en estos casos se puede utilizar el examen patológico intraoperatorio. Conclusiones: El bazo ectópico en parénquima testicular es una entidad rara y de difícil diagnóstico. Es importante considerar esta condición como un diagnóstico diferencial en los trastornos testiculares, especialmente en el caso de la hipótesis diagnóstica de cáncer testicular, para que la extracción del testículo pueda ser evitada mediante el examen patológico intraoperatorio.

ABSTRACT Extraovarian granulosa cell tumors (GCTs) develop from ectopic gonadal tissue situated along the embryonal route of the genital ridge. Primary retroperitoneal tumors are extremely rare, with an incidence of 02% -06% and 80-85% probability of malignancy. Only eight such case reports have been published previously. We herein, report a rare case of extraovarian retroperitoneal GCT in a 55-year-old woman who presented with intermittent left lumbar region pain of one-year duration. She had a history of hysterectomy and bilateral salpingo-oophorectomy 8 years ago for uterine leiomyoma. Laparotomy revealed a retroperitoneal mass measuring 8cm x 10cm x 20cm in size, solid cystic with areas of necrosis and hemorrhage. The gross features, classical histopathology, and positive immunostaining of the retroperitoneal mass with inhibin, calretinin, PR, WT1 and immunonegativity for EMA were characteristic of adult-type GCT. Excluding any previous history of primary ovarian GCT in this patient, a de-novo retroperitoneal diagnosis was established.

Abstract 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. (BRB GRB (GRB biodiversity Nature 38500 38 500 38,50 extinction 138300 138 300 138,30 zoos universities organizations years grafting infertility recover cryopreserve speciesspecific specific protocols activities peccaries agoutis cavies Additionally close 3850 3 50 38,5 13830 13 30 138,3 385 5 38, 1383 1 138,

Abstract: In this study we tried to reveal if under conditions of high-stress mussels presented some trade off about reproductive aspects like size/frequency of oocytes and the reproductive cycle of the mussel Brachidontes rodriguezii. This species dominates the rocky intertidal coast of Buenos Aires province, Argentine and is a prey item of several exploited species. Mussels were collected from December 2011 to May 2013 in the upper level of it distribution in the high intertidal coast. Gonad tissue samples were processed according to standard histological procedures. Four gonadal development stages were described for males and females all year round. According to the significant decrease in the frequency of mature oocytes, the spawning events were in February 2012/2013. This study also reports hermaphrodite and parasite mussels in very low frequency. We concluded that the frequency and duration of the post-spawning gonadal development stage were the most notable difference along the vertical distribution, suggesting that the high-stress condition of high intertidal impacts negatively on gonad restoration.

Resumen: En este estudio evaluamos si bajo condiciones de alto estrés los mejillines Brachidontes rodriguezii presentaban algún compromiso en aspectos reproductivos como el tamaño/frecuencia de los ovocitos y el ciclo reproductivo. Esta especie domina el intermareal rocoso de la provincia de Buenos Aires, Argentina y es presa de varias especies explotadas. Los mejillines se recolectaron desde diciembre de 2011 hasta mayo de 2013 en el nivel superior del intermareal costero. Se procesaron muestras de tejido gonadal según procedimientos histológicos estándar. Se describieron cuatro etapas de desarrollo gonadal para machos y hembras durante todo el año. De acuerdo con la disminución significativa en la frecuencia de ovocitos maduros, los eventos de desove fueron en febrero de 2012/2013. Este estudio también reporta mejillines hermafroditas y parásitos en muy baja frecuencia. Concluimos que la frecuencia y la duración de la etapa de desarrollo gonadal postdesove resultó ser la diferencia más notable a lo largo de la distribución vertical, sugiriendo que la condición de alto estrés del intermareal superior impacta negativamente en la restauración gonadal.

SUMMARY: Studies have shown the adverse effects of epileptic seizures on reproductive health. The aim of the present study was to investigate morphological changes, apoptosis and GABA localization in the testis tissue of genetic absence epilepsy rats. Testis tissues of GAERS and Wistar rats were processed for paraffin embedding and electron microscopy. Sections were stained with hematoxylin and eosin, Masson’s trichrome and periodic acid-Schiff reaction. GABA immunohistochemistry was applied for determining the alterations in GABA levels. GABA immunoreactivity was observed in the seminiferous tubules and interstitial areas of both GAERS and Wistar rats. GABA immunoreactivity was found to be decreased in GAERS compared to Wistar group. Electron microscopic observations showed that GABA was present in manchette microtubules, sperm tail and neck at different phases of spermiogenesis. Qualitative observations revealed that testis tissues of GAERS showed reduced sperm in the seminiferous tubules compared to the Wistar controls. In conclusion, we demonstrated GABAergic system in the seminiferous tubules of control and GAERS rats, in parallel with the previous studies; and there were alterations in this system in GAERS. We suggest that these alterations in absence epilepsy may also affect the gonadal system, resulting in decreased sperm production.

RESUMEN: Los estudios han demostrado los efectos adversos de las convulsiones epilépticas sobre la salud reproductiva. El objetivo del presente estudio fue investigar los cambios morfológicos, la apoptosis y la localización de GABA en el tejido testicular de ratas con epilepsia de ausencia genética. Se procesaron tejidos testiculares de ratas GAERS y Wistar para inclusión en parafina y microscopía electrónica. Las secciones se tiñeron con hematoxilina y eosina, tricrómico de Masson y reacción de ácido peryódico de Schiff. Se aplicó inmunohistoquímica de GABA para determinar las alteraciones en los niveles de GABA. Se observó inmunorreactividad de GABA en los túbulos seminíferos y las áreas intersticiales de las ratas GAERS y Wistar. Se encontró que la inmunorreactividad de GABA estaba disminuida en GAERS en comparación con el grupo Wistar. Las observaciones microscópicas electrónicas mostraron que GABA estaba presente en los microtúbulos, la cola y el cuello del espermatozoide en diferentes fases de la espermiogénesis. Las observaciones cualitativas revelaron que los tejidos testiculares de GAERS mostraron una reducción de los espermatozoides en los túbulos seminíferos en comparación con los controles Wistar. En conclusión, demostramos el sistema GABAérgico en los túbulos seminíferos de ratas control y GAERS, en paralelo con estudios previos; y además se observaron alteraciones en este sistema en GAERS. Sugerimos que estas alteraciones en epilepisa de ausencia genética también pueden afectar el sistema gonadal, resultando en una disminución de la producción de semen.

Resumen Objetivos: La etiología de la disgenesia gonadal y el síndrome ovotesticular se desconoce en la mayoría de los casos. Para realizar la caracterización citogenética y molecular de un grupo de pacientes con síndrome ovotesticular y disgenesia gonadal completa a partir de muestras de sangre periférica y tejido gonadal. Material y métodos: Se incluyeron un total de 6 pacientes, 3 con diagnóstico de síndrome ovotesticular 46, XX, uno diagnosticado con 46, XY síndrome ovotesticular; uno con sospecha de disgenesia gonadal 46, XX y otro con disgenesia gonadal completa 46, XY. Resultados: Todos los pacientes fueron evaluados con cariotipo, hibridación in situ fluorescente (FISH) para SRY, amplificación de sonda dependiente de ligación múltiple (MLPA) e hibridación genómica comparativa (aCGH) en muestras de sangre periférica. En los casos con tejido gonadal disponible, los niveles de expresión genética de SOX3, SRY y SOX9 se determinaron mediante PCR en tiempo real e inmunofluorescencia. Se descartaron reordenamientos relacionados con el gen SRY. No se detectaron deleciones/duplicaciones o variaciones en el número de copias (NVC) como etiología del trastorno del desarrollo sexual en ninguno de los pacientes estudiados. En un caso de síndrome ovotesticular 46, XX, el cariotipo gonadal era diferente del cariotipo en sangre periférica. Se observó expresión aberrante de SOX3 y SOX9 en tejido gonadal de un caso con síndrome ovotesticular 46, XX. Conclusiones: Se documentaron niveles más bajos de expresión de SRY y SOX9 en comparación con los niveles en líneas celulares humanas de testículo embrionario y Sertoli en el tejido gonadal de un caso con síndrome ovotesticular 46, XY. Los estudios citogenéticos y moleculares de las gónadas como complemento del estudio de sangre periférica tienen el potencial de enriquecer la comprensión de los trastornos del desarrollo sexual en pacientes que son XX o XY en sangre periférica.

Abstract Objectives: The etiology of gonadal dysgenesis and ovotesticular syndrome is unknown in a majority of cases. The aim of the study was to perform cytogenetic and molecular characterization of a group of patients with ovotesticular syndrome and complete gonadal dysgenesis from peripheral blood and gonadal tissue samples. Materials and methods: A total of 6 patients were included, 3 with diagnosis of 46,XX ovotesticular syndrome, one diagnosed with 46,XY ovotesticular syndrome; one with suspected 46,XX gonadal dysgenesis, and one with 46,XY complete gonadal dysgenesis. Results: All patients were evaluated with karyotype, fluorescent in situ hybridization (FISH) for SRY, multiplex ligation-dependent probe amplification (MLPA) and comparative genomic hybridization (aCGH) on peripheral blood samples. In cases with available gonadal tissue, the levels of genetic expression of SOX3, SRY, and SOX9 were determined through real-time PCR and immunofluorescence. Rearrangements involving SRY gene were ruled out. No deletions/duplications or copy-number variations (CNV) were detected as the etiology for sexual development disorder in any of the patients studied. In a case of 46,XX ovotesticular syndrome, the gonadal karyotype was different from the karyotype in peripheral blood. Aberrant expression of SOX3 and SOX9 in gonadal tissue of a case with 46,XX ovotesticular syndrome was observed. Conclusions: Lower levels of expression of SRY and SOX9 compared to the levels in human cellular lines of embryonic testicle and Sertoli were documented in the gonadal tissue of a case with 46,XY ovotesticular syndrome. Cytogenetic and molecular studies of the gonads as a complement to peripheral blood study have the potential to enrich the understanding of sexual development disorders in patients who are XX or XY in peripheral blood.

Abstract Objective: Disorders of sexual development are a group of congenital diseases that affect the normal formation of genital structures. Within the pathophysiologic mechanisms described, there are genetic factors caused by chromosomal or sex-determining gene alterations. Therefore, chromosomal analysis is an essential priority in the diagnostic approach. Alterations in the chromosomes and the SRY gene as a cause of disorder of sexual development was analyzed herein. Material and methods: G or R-banding karyotype and FISH analyses for the SRY gene were performed in lymphocytes, gonadal tissue, and scrotal tissue in twelve cases, three cases, and one case, respectively. The clinical information was obtained from the patients’ medical reports. Results: In 9 (73%) cases, the assigned sex was male, and in 3 (27%) cases, it was female. Karyotype 46,XY was found in 8 (66%) cases, 46,XX in 2 (17%) cases, and mosaic karyotype in 2 (17%) cases with idic(Y). A single case of gonadal tissue showed mosaicism due to the presence of a tetraploid cell line. The most common clinical diagnosis was abnormal genital differentiation in 8 (67%) cases, followed by hypospadias in 5 (41.7%) cases. Conclusions: The results show the importance of applying different cytogenetic tests in making the diagnosis, and the need for a multidisciplinary team to address the disorder.

Resumen Objetivo: Los trastornos del desarrollo sexual son un grupo de enfermedades congénitas que afectan la formación normal de los genitales. Dentro los mecanismos fisiopatológicos descritos existen factores genéticos causados por alteraciones cromosómicas o en los genes determinantes en la diferenciación sexual. En este trabajo se analizaron alteraciones cromosómicas y en el gen SRY como posible causa del trastorno. Se realizó cariotipo con bandas G o R y FISH para SRY en linfocitos, tejido gonadal y tejido escrotal. Materiales y métodos: La información clínica de los sujetos de investigación se obtuvo de los informes de los médicos tratantes. En 9 (73%) casos el sexo asignado fue masculino y en 3 (27%) casos fue femenino. 8 de los casos (66%) tuvieron cariotipo 46,XY; 2 casos (17%) 46,XX y en 2 casos (17%) se reportaron mosaicos con presencia de idic(Y). Un solo caso de tejido gonadal mostró mosaicismo debido a la presencia de una línea celular tetraploide. El diagnóstico clínico más frecuente fue de genitales ambiguos en 8 casos (67%). Seguido de hipospadias en 5 casos (41,7%). Conclusiones: Los resultados muestran la importancia de aplicar diferentes pruebas citogenéticas en el diagnóstico y la necesidad del seguimiento de los pacientes por un equipo transdisciplinario para abordar estas condiciones clínicas.

RESUMO: Embora Trachemys scripta elegans seja uma espécie exótica popular como animal de estimação no Brasil, estudos sobre biologia e capacidade reprodutivas são inexistentes no Cerrado brasileiro. Este estudo analisou características ovarianas e do oviduto e a capacidade de produção de ovos em T. scripta elegans criadas neste bioma, correlacionando estes achados ao tamanho dos espécimes e a maturidade sexual, visando comparações com populações nativas e exóticas, bem como interespecíficas e contribuir para a compreensão de seu impacto nos ecossistemas invadidos e com o estabelecimento de programas de erradicação. Assim, 39 fêmeas tiveram avaliadas a biometria corporal e a morfologia e morfometria dos ovários e ovidutos. De acordo com o tamanho dos folículos ovarianos as fêmeas foram separadas em G1 (N= 10): com folículos Classe I (>5-10 mm), G2 (N= 20): com folículos Classe I e Classe II (>10-25 mm) e G3 (N= 9) com folículos Classe I, Classe II e Classe III (>25 mm). À análise de variância, teste de Scott-Knott e à análise de correlação de Pearson verificou-se que não houve diferença significativa entre os grupos na biometria corporal; no índice gonadossomático médio e na morfometria gonadal, apenas a largura dos ovidutos no antímero direito e a massa e a largura no antímero esquerdo foram maiores no G3, o único que apresentou ovos. Houve desenvolvimento positivo e harmônico entre massa corporal, carapaça e plastrão e o crescimento gonadal ocorreu concomitante ao crescimento corporal, indicando maior potencial reprodutivo e relação positiva entre o tamanho da ninhada de ovos e o da fêmea. O índice gonadossomático mostrou-se um bom indicador reprodutivo e a avaliação ovariana um melhor indicador da maturidade sexual que o comprimento máximo da carapaça. Ovários foram estruturas irregulares, sem delimitação entre a região cortical e medular e repletos de folículos vitelogênicos de diferentes diâmetros, folículos atrésicos e corpos lúteos, que refletiram a complexidade ovariana da espécie e a presença de hierarquia folicular. No estroma escasso foram observados dois leitos germinativos por ovário e a presença de lacunas muito próximas aos folículos e associadas aos vasos sanguíneos. A análise do tecido gonadal revelou três tipos de oócitos de acordo com as características do citoplasma: homogêneo, vesicular ou vesicular no córtex com grânulos aparentes. Ovidutos eram funcionais e separados, unindo-se apenas na porção final para formar a cloaca e subdividiam-se em infundíbulo, tuba uterina, istmo, útero e vagina. A estrutura da tuba uterina era constituída de serosa, muscular e mucosa, a qual era repleta de glândulas. A presença de ovos nos ovidutos indicou que os espécimes podem se reproduzir no cerrado brasileiro. Este estudo fornece informações básicas e relevantes da biologia e capacidade reprodutivas de T. scripta elegans no Cerrado brasileiro e pode contribuir com a compreensão de seu impacto nos ecossistemas invadidos e com o estabelecimento de programas de erradicação, uma vez que a extração de fêmeas com capacidade reprodutiva pode contribuir com a diminuição do rendimento reprodutivo anual da espécie e diminuir seu efeito sobre a biodiversidade local.

ABSTRACT: Although Trachemys scripta elegans is an exotic species popular as a pet in Brazil, studies on reproductive biology and capacity are non-existent in the Brazilian Cerrado. This study analyzed ovarian and oviduct characteristics and the egg production capacity of T. scripta elegans grown in this biome. The findings will associate with the size of the specimens and the sexual maturity, aiming at comparisons with native and exotic populations, as well as interspecific and contributing to the understanding of its impact on the invaded ecosystems and the establishment of eradication programs. Thus, 39 females had evaluated the body biometry and the morphology and morphometry of the ovaries and oviducts. G2 (N=20): with Class I (>5-10mm) follicles, with Class I and Class II (>10-fold) follicles, 25mm) and G3 (N=9) with Class I, Class II and Class III (>25mm) follicles. Analysis of variance, Scott-Knott’s test, and Pearson’s correlation analysis showed that there was no significant difference between the groups in body biometry; in the mean gonadosomatic index and gonadal morphometry, only the width of the oviducts in the right antimer and the mass and width in the left antimer were higher in G3, the only one that presented eggs. There was positive and harmonic development between body mass, carapace, and plastron, and gonadal growth occurred concomitantly with body growth, indicating a higher reproductive potential and a positive relationship between the size of the litter and the female litter. The gonadosomatic index proved to be an excellent reproductive indicator, and the ovarian evaluation was a better indicator of sexual maturity than the maximum carapace length. Ovaries were irregular structures, without delimitation between the cortical and medullary regions and filled with vitelogenic follicles of different diameters, atresic follicles, and corpora lutea, which reflected the ovarian complexity of the species and the presence of follicular hierarchy. In the scarce stroma, two germinative beds were observed per ovary and the presence of gaps very close to the follicles and associated with the blood vessels. Analysis of gonadal tissue revealed three types of oocytes according to cytoplasmic characteristics: homogeneous, vesicular or vesicular in the cortex with apparent granules. Oviducts were functional and separated, joining only in the final portion to form the cloaca and subdivided into infundibulum, tuba, isthmus, uterus, and vagina. The structure of the uterine tube was composed of serosa, muscular and mucous, which was full of glands. The presence of eggs in the oviducts indicated that the specimens can reproduce in the Brazilian Cerrado. This study provides necessary and relevant information on the reproductive biology and capacity of T. scripta elegans in the Brazilian Cerrado and can contribute to the understanding of its impact on the invaded ecosystems and the establishment of eradication programs. The extraction of females with capacity can reduce the annual reproductive yield of the species and decrease its effect on local biodiversity.

ABSTRACT. Triploid fish are usually sterile. Thus, the energy and nutrients intended for sexual maturation may be available to enhance flesh quality and physical growth. The present study aimed to investigate differences in the metabolic substrates, lipids and proteins, between storage tissues from diploid and triploid female rainbow trout. Monthly, metabolic substrates were quantified in liver, muscle, and ovaries, which were collected during the first reproductive cycle. In general, it was possible to identify a seasonal and similar deposition of metabolites in different tissues of 2n and 3n females, mainly at early stages of gonadal maturation. However, from the stages 5-6, the ovaries showed great differences between ploidies, with higher concentration of lipids and protein in 2n females. This result reflects the incorporation of vitellogenin in oocytes, which is a process that does not occur in 3n females. It was possible to observe seasonal hepato-somatic index changes in 2n females, with higher values observed in the post-ovulatory stage, and the triploid animals showed lower values compared to 2n, with no seasonal difference. Viscero-somatic index can reflect the mobilization of substrates, with higher values found for 2n females in stage 5-6, which is the period of active mobilization of tissue substrates.

Resumo: O hábito alimentar de peixes associado à morfologia do tubo digestivo pode fornecer informações biológicas e ecológicas úteis. Entre janeiro e dezembro de 2012, examinamos 16 espécies de peixes descartadas por pesca de arrasto na Bacia Potiguar, nordeste do Brasil, e as classificamos em quatro grupos, de acordo com o Índice Alimentar-IAi. Nossos objetivos foram: (1) fornecer informações sobre a composição da dieta de uma espécie representante de cada grupo (Opisthonema oglinum, Conodon nobilis, Larimus breviceps e Sphoeroides testudineus), (2) examinar a morfologia de seu tubo digestivo e (3) associar os itens alimentares ingeridos com as características morfológicas descritas. O conteúdo estomacal foi coletado e classificado de acordo com o IAi nas categorias: principal, acessória e secundária; fragmentos de esôfago, estômago e intestino delgado foram examinados pela técnica de secção em parafina. Os dados biométricos (comprimento total-CT, comprimento padrão-CP e peso total-PT), peso do estômago-PE e peso gonadal-PG foram registrados e aplicados em dois índices: (1) Índice de Repleção Estomacal - IR (IR = PE / PT × 100) e (2) Índice Gonadossomático - IGS (IGS = PG / PT × 100) para análises de atividade alimentar e reprodutiva, respectivamente. As principais categorias alimentares foram: (1) Teleostei para C. nobilis, (2) Crustacea para L. breviceps e O. oglinum, e (3) Mollusca para S. testudineus. Maior atividade alimentar ocorreu durante a estação seca, e o IGS foi maior na estação chuvosa. O exame morfológico revelou estrutura regular dos tecidos analisados, destacando-se a presença de glândulas gástricas tubulares e camadas musculares espessas no estômago (O. oglinum; L. breviceps); cecos pilóricos bem desenvolvidos no intestino (O. oglinum; C. nobilis; L. breviceps); e presença de pseudoestômago (S. testudineus). Nosso estudo confirma que o tubo digestivo das espécies examinadas possui adequação morfológica ao hábito alimentar carnívoro e fornece informações úteis sobre as estruturas dos tecidos associados à composição da dieta.

Abstract: Fish feeding habit associated to the morphology of their digestive tube can provide useful biological and ecological information. Between January and December 2012, we examined 16 fish species discarded by trawl in the Potiguar basin, northeastern Brazil, and classified them into four groups according to Alimentary Index - IAi. Our aims were to: (1) provide information on diet composition of one species represented by each group (Opisthonema oglinum, Conodon nobilis, Larimus breviceps and Sphoeroides testudineus), (2) examine the morphology of their digestive tube, and (3) associate ingested food items with the morphological features. Stomach contents were collected and classified according to IAi: main, accessory and secondary categories; fragments of esophagus, stomach, and small intestine were examined using paraffin section technique. Biometric data (total length-TL, standard length-SL and total weight-TW), stomach weight-SW, and gonadal weight-GW were recorded and applied in two indices: (1) the stomach Repletion Index - SRI (SRI = SW/TW × 100) and (2) the gonadosomatic index - GSI (GSI = GW/TW × 100) for the analyses of feeding and reproductive activities, respectively. The main food categories were (1) Teleostei for C. nobilis, (2) Crustacea for L. breviceps and O. oglinum, and (3) Mollusca for S. testudineus. The feeding activity was higher during the dry season, while the GSI was higher in the rainy season. The morphological analysis revealed a regular structure of the tissues analyzed. We highlight the presence of tubular gastric glands and strong muscular walls in the stomach (O. oglinum; L. breviceps); well-developed pyloric caecum in the intestine (O. oglinum; C. nobilis; L. breviceps); and presence of pseudo-stomach (S. testudineus). Our study confirms that the digestive tube of the examined species has morphological adequacy for carnivorous feeding habit, providing useful information on tissue structures associated to diet composition.