Effect of laser-assisted hatching on birth weight in blastocyst cultureof low-grade cleavage embryos / 预防医学

Objective@#To examine the impact of laser-assisted hatching (LAH) technique on perinatal outcomes in blastocyst culture of low-grade cleavage embryos, so as to provide insights into improving the utilization of low-grade cleavage embryos and embryo quality.@*Methods@#A total of 369 single live births after transfer of thawed blastocysts following in vitro fertilization or intracytoplasmic sperm injection at Zhejiang Provincial People's Hospital were selected as subjects, and they were divided into 51 conventional culture blastocysts and 318 LAH blastocysts based on whether LAH was performed on day 4 of blastocyst culture. Gestational age, birth weight, birth defects and maternal perinatal information were collected, and the prevalence of premature birth, birth weight and birth defects were analyzed after propensity score matching (PSM).@*Results@#After PSM, 98 matched cases were included in the study. There were no statistically significant differences in maternal age, body mass index, type of infertility and blastocyst age (P>0.05), indicating effective matching. The birth weight of offspring in the LAH group was lower than that in the conventional culture group [(3 261.08±432.24) g vs. (3 464.18±444.46) g; P<0.05].@*Conclusion@#The birth weight of offspring can be reduced by using LAH during blastocyst culture of low-grade cleavage embryos.

Anidación de la tortuga marina Lepidochelys olivacea (Testudinata: Cheloniidae) en Campamento Tortuguero Cedeño, Honduras (2011-2021) / Nesting of olive ridley sea turtle Lepidochelys olivacea (Testudinata: Cheloniidae) in Campamento Tortuguero Cedeño, Honduras (2011-2021)

Introducción: El Campamento Tortuguero de Cedeño ha sido el sitio menos investigado del Golfo de Fonseca, donde se protege a la tortuga golfina en Honduras desde 1975. Objetivo: Evaluar la anidación de la tortuga Golfina (Lepidochelys olivacea) durante la temporada de veda entre el 2011 y 2021 en Campamento Tortuguero Cedeño, Choluteca, Honduras. Métodos: Entre 2011 a 2021, se llevó a cabo el monitoreo diario de las actividades de anidación durante la veda del 1 al 25 de septiembre. Los patrullajes se realizaron entre las 6:00-18:00 h, y las 18:00-5:00 h. Se registró el número total de tortugas que anidaban y se recogieron sus huevos, que se transportaron al criadero, donde se tabularon los resultados de las puestas y las crías. Resultados: Se registró un total 1 065 tortugas de L. olivacea, 95 051 huevos recolectados, 1 065 nidos marcados en tres playas que fueron reubicados en viveros artificiales y una eclosión exitosa de 62 747 neonatos. La playa Las Doradas fue el sitio con el mayor número de tortugas anidadoras, seguido de Los Delgaditos y por último Cedeño. El promedio de la frecuencia de anidación fue de 96 nidos. Del 2011 al 2021 el esfuerzo de recolección de los nidos aumentó en un 91.6 %, pasando de 84 a 161 nidos. El número de personas patrullando se asoció con la cantidad de nidos detectados en las playas. Conclusión: Los esfuerzos de monitoreo y conservación para la especie han indicado que ha habido un incremento en la anidación de L. olivacea en las tres playas, con un mayor incremento en Playa Las Doradas. Este escenario comprueba la funcionalidad de la veda en esta zona.

Introduction: The Cedeño Turtle Camp has been the least researched site in the Fonseca Gulf, where Olive Ridley Turtles in Honduras have been protected since 1975. Objective: To evaluate the nesting of Olive Ridley turtles (Lepidochelys olivacea) during the closed season from 2011 to 2021 in Campamento Tortuguero Cedeño, Choluteca, Honduras. Methods: From 2011 to 2021, daily monitoring of nesting activities was conducted during the closed season from the 1st to 25th of September. Patrols were conducted between 6:00-18:00 h, and 18:00-5:00 h. The total number of nesting turtles was recorded, and their eggs were collected and transported to the hatchery, where clutch and hatchling performance were tabulated. Results: A total of 1 065 L. olivacea turtles were recorded, 95 051 eggs collected, 1 065 nests marked on three beaches that were relocated in artificial hatcheries and a successful hatching of 62 747 hatchlings. Las Doradas beach was the site with the highest number of nesting turtles, followed by Los Delgaditos and lastly Cedeño. The average nesting frequency was 96 nests. From 2011 to 2021 the nest collection effort increased by 91.6 %, from 84 to 161 nests. The number of people patrolling was associated with the number of nests detected on the beaches. Conclusion: Monitoring and conservation efforts for L. olivacea in the Campamento Tortuguero Cedeño show a positive trend in nesting with a greater increase in Playa Las Doradas. This scenario proves the functionality of the closed season in this area.

Immunochemical studies of Toxocara canis proteases

@#Proteases of nematodes play a crucial role in larval molting and, in addition to their active role in egg hatching, proteases are also considered a crucial factor in tissue invasion and connective tissue remodeling. In Toxocara canis, proteases play important roles throughout the complex life cycle. They can degrade components of a model of extracellular matrix, basement membranes and different physiological substrates. In the present study, measurements of the proteolytic activity of the perivitelline fluid (PF) surrounding Toxocara canis embryos at different stages of development, the hatching fluid (HF) surrounding the infective larvae, as well as the excretory secretory (ES) products of the larvae in the culture media were performed. Measurements were made using casein as substrate following the Sigma non-specific protease activity assay. The results showed that enzyme activity increased as the embryo matured. The infective larvae were found to continuously produce proteases in the surrounding HF and ES products after in vitro cultivation indicating that Toxocara canis proteases might be important for the worm in the egg and the host. Optimal enzymatic activity was found at pH 8. Incubation of the antiserum from infected mice with the HF and ES products decreased their proteolytic activities, suggesting that there may be a link between the proteases present in these fluids and the immune response.

Efecto de la reubicación de nidos en el éxito reproductivo de la tortuga marina Lepidochelys olivacea (Testudinata: Cheloniidae) / Effect of nest relocation on the reproductive success of the marine turtle Lepidochelys olivacea (Testudinata: Cheloniidae)

Resumen Introducción: Para incrementar el número de crías de tortugas marinas, es necesario mejorar estrategias de conservación, como incubación de nidos, en condiciones de vivero que contrarresten los efectos de las condiciones climáticas extremas. Objetivo: Comparar cinco parámetros de éxito reproductivo (éxito de eclosión, huevos sin desarrollo aparente, crías muertas, mortalidad embrionaria y período de incubación) de la tortuga marina Lepidochelys olivacea. Métodos: Se evaluó la incubación de nidos bajo dos períodos de reubicación de nidos (P1 ~28.8 °C agosto-septiembre y P2 ~27.1 °C octubre-noviembre, temperatura ambiental promedio, bajo condiciones de vivero, en el 2018 en Guerrero, México. Resultados: Se encontraron diferencias significativas en el éxito de eclosión y la mortalidad embrionaria entre los períodos de reubicación de nidos (P < 0.001). En P1, se observaron valores más bajos de éxito de eclosión (77.0 % P1 vs 88.6 % P2) y valores más altos de mortalidad embrionaria (13.7 % P1 vs 3.3 % P2) comparados con los de P2. Es importante señalar que, en el presente estudio la mortalidad embrionaria se presentó en una etapa tardía de desarrollo, es decir, en embriones casi completamente desarrollados. No se encontraron diferencias entre periodos respecto a los parámetros de huevos sin desarrollo aparente, crías muertas y período de incubación. Conclusiones: Las diferencias entre las condiciones ambientales durante el primer período de reubicación de nidos en comparación con el segundo período parecen afectar el éxito de eclosión y mortalidad embrionaria de L. olivacea. Por lo tanto, es importante tomar medidas al respecto durante esta etapa de incubación para tratar de mejorar la incubación de nidos bajo condiciones de vivero.

Abstract Introduction: In order to increase the number of sea turtle hatchlings, it is necessary to improve conservation strategies, such as nest incubation in hatchery conditions that counteract the effects of extreme climatic conditions. Objective: To compare five reproductive success parameters (hatching success, eggs with no apparent embryonic development, dead hatchlings, embryonic mortality, and incubation period) of the sea turtle Lepidochelys olivacea. Methods: The incubation of nests was evaluated under two periods of nest relocation, P1 ~28.8 °C (August and September) and P2 ~27.1 °C (October and November), average ambient temperature, under hatchery condition, in 2018 in Guerrero, Mexico. Results: Significant differences were found in hatching success and embryonic mortality between nest relocation periods (P < 0.001). In P1, lower values of hatching success (77.0 % P1 vs 88.6 % P2) and higher values of embryonic mortality (13.7 % P1 vs 3.3 % P2) were observed compared to those of P2. It's important pointing that, in the present study, embryonic mortality occurred at a late stage of development, that is, in almost fully developed embryos. No differences were found between periods regarding the parameters eggs with no apparent embryonic development, dead hatchlings and incubation period. Conclusions: The differences between the environmental conditions during the first period of nest relocation compared to the second period, respectively, seem to affect the hatching success and embryonic mortality of L. olivacea. Therefore, it is important to take action on this issue during this incubation stage to try to improve the incubation of nests under hatchery conditions.

Nesting trends of olive ridley sea turtles Lepidochelys olivacea (Testudinata: Cheloniidae) on two beaches in Northwestern Mexico after 30 and 40 years of conservation / Tendencias de anidación de la tortuga golfina Lepidochelys olivacea (Testudinata: Cheloniidae) en dos playas del noroeste de México después de 30 y 40 años de conservación

Abstract Introduction: Although olive ridley sea turtle (Lepidochelys olivacea) are the most abundant sea turtles in the world, quantitative information is scarce and unevenly distributed among regions. There are many management and conservation programs for this species, and assessments are necessary to identify nesting trends and effectively manage current conservation programs. PROTORMAR-UAS is a Research and Conservation program for the olive ridley turtle created by the Autonomous University of Sinaloa, Mexico. The program utilizes two biological stations: Santuario Playa Ceuta (SPC) and Playa Caimanero (PC). Objective: To evaluate the nesting trend of olive ridley turtles on two beaches in Northwestern Mexico and to predict prospective nesting trends for the next 30 years. Methods: Using annual nesting data collected over 40 years at SPC (1976-2016) and 30 years at PC (1986-2016), we evaluated nesting trends, hatching success, predation and poaching of olive ridley turtles on the two beaches in Northwestern Mexico. Then, prospective nesting estimates for the next 30 years were calculated predictive time series model. Results: A positive and significant correlation was identified between the number of annual nests and time for both beaches (rho = 0.850, P ≤ 0.01 for SPC; rho = 0.677, P ≤ 0.01 for PC); the average hatching success rates were 65.09 at SPC and 60.72 % at PC. The predictive time-series model indicated that the numbers of nests will continue to increase through 2045, increasing three-fold at SPC and six-fold at PC with respect to the last year of monitoring. Conclusions: There was a clear positive trend in the number of olive ridley sea turtle nests at both sites, which is consistent with trends found in other recent studies from the region. Therefore, we suggest that PC be designated a legally protected nesting area since it is located within the latitudinal limits of olive ridley nesting and given the need for resources for camp operation considering increased nesting and current problems with predation and poaching. Because in Mexico operating a nesting beach without any protection status implies not having a budget for its management.

Resumen Introducción: A pesar de que las tortugas golfinas (Lepidochelys olivacea) son las tortugas marinas más abundantes del mundo, su información cuantitativa disponible es escasa y se encuentra distribuida de manera desigual entre regiones. Existen muchos programas de manejo y conservación para esta especie, y sus evaluaciones son necesarias para identificar tendencias de anidación y poder manejar de manera efectiva los programas de conservación actuales. PROTORMAR-UAS es un programa de Investigación y Conservación de la tortuga golfina creado por la Universidad Autónoma de Sinaloa, México. El Programa cuenta con dos estaciones biológicas: Santuario de Playa Ceuta (SPC) y Playa Caimanero (PC). Objetivo: Evaluar la tendencia de anidación de la tortuga golfina en dos playas del noroeste de México y predecir las tendencias prospectivas de anidación para los próximos 30 años. Métodos: A partir de los datos de registros anuales de anidación de 40 años para SPC (1976-2016) y 30 años para PC (1986-2016), evaluamos las tendencias de anidación, el éxito de la eclosión y los problemas de depredación y saqueo de nidos de la tortuga golfina en las dos playas del noroeste de México. Posteriormente, se calcularon las estimaciones prospectivas de anidación para los próximos 30 años usando un modelo predictivo de series de tiempo. Resultados: Se identificó una correlación positiva y significativa entre el registro anual de nidos y el tiempo de estudio para ambas playas (rho = 0.850, P ≤ 0.01 para SPC; rho = 0.677 y P ≤ 0.01 para PC); así como el éxito de eclosión promedio de 65.09 para SPC y de 60.72 % para PC. El modelo predictivo de series de tiempo indicó que las anidaciones continuarán aumentando para el 2045, tres veces para SPC y seis para PC, con respecto al último año de monitoreo. Conclusiones: Hay una clara tendencia positiva de anidación de la tortuga golfina en ambos sitios, lo cual es consistente con la tendencia observada en otros estudios recientes de la región. Por lo tanto, sugerimos incluir a PC como un área de anidación legalmente protegida, la cual se ubica en los límites latitudinales de anidación de la tortuga golfina, dada la necesidad de contar con recursos disponibles para la operación del campamento ante el aumento de anidaciones y de problemas de depredación y saqueo. Porque en México operar una playa de anidación sin ningún estatus de protección implica no tener presupuesto para su manejo.

The spawning method affects the reproductive efficiency of piracanjuba fish (Brycon orbignyanus) in restocking programs / El método de desove afecta la eficiencia reproductiva de piracanjuba (Brycon orbignyanus) en programas de repoblación / O método de desova afeta a eficiência reprodutiva de piracanjuba (Brycon orbignyanus) em programas de repovoamento

Abstract Background: Preserving the genetic diversity of wild fish is an important consideration for restocking programs, as inbreeding can compromise progeny survival as well as impact the resilience of natural populations. Objective: To evaluate the influence of spawning method: semi-natural (SN) or strip-spawning (ST), and the number of breeders (1♀:3♂ and 2♀:6♂) on the reproductive efficiency and genetic diversity of B. orbignyanus progeny destined for restoration of wild stocks. Methods: Rates of fertilization, hatching and broodfish mortality were recorded. For genetic evaluations (allele frequency, observed and expected heterozygosity, Shannon index, inbreeding coefficient, molecular variance analysis, and genetic differentiation), breeders (n=24), and their progenies (90 larvae/treatment) were sampled and analyzed using eight microsatellite markers. Results: Higher fertilization and hatching rates, and lower broodfish mortality were observed for the SN method (p<0.05), whereas the number of breeders did not affect these parameters (p>0.05). Interaction between spawning method and number of breeders was not significant (p>0.05). The amplified microsatellite loci produced a total of 30 alleles, with sizes between 80 and 225 bp and their frequencies indicated an increase (p<0.05) of genetic diversity in the progenies, but low genetic differentiation between treatments (p>0.05). Conclusion: The spawning methods and number of breeders tested increased equally the genetic diversity of the progeny, with low genetic differentiation between treatments. In contrast, rates of fertilization, hatching and brood fish mortality revealed that the SN method resulted in the best reproductive efficiency due to the handling stress and injuries caused by ST. Thus, SN proves to be the most suitable spawning-method for B. orbignyanus in restocking programs.

Resumen Antecedentes: Mantener la diversidad genética de los peces salvajes es una consideración importante para los programas de repoblación, ya que la endogamia puede comprometer la supervivencia de la progenie y afectar la supervivencia de las poblaciones naturales. Objetivo: Evaluar la influencia del método de desove (seminatural - SN o en franjas - ST) y el número de reproductores (1♀:3♂ y 2♀:6♂) sobre la eficiencia reproductiva y la diversidad genética de progenies de B. orbignyanus destinados a la restauración de poblaciones silvestres. Métodos: Se monitorearon las tasas de fertilización, eclosión y mortalidad de reproductores. Para las evaluaciones genéticas (frecuencias alélicas, heterocigosidad observada y esperada, índice de Shannon, coeficiente de endogamia, análisis de varianza molecular y diferenciación genética) los reproductores (n=24) y su progenie (90 larvas/tratamiento) se muestrearon y analizaron utilizando ocho marcadores microsatélites. Resultados: La interacción entre el método de desove y el número de reproductores no fue significativa (p>0,05). Se obtuvieron mejores tasas de fecundación y eclosión (p<0,05), y una menor mortalidad de reproductores (p<0,05) con el método SN, mientras que el número de reproductores no tuvo efecto (p>0,05). Los loci de microsatélites amplificados produjeron un total de 30 alelos con tamaños entre 80 y 225 pb, y sus frecuencias indicaron un aumento (p<0,05) en la diversidad genética de las progenies, pero una baja diferenciación genética entre tratamientos (p>0,05). Conclusión: Los métodos de desove y el número de reproductores evaluados aumentaron de la misma manera la diversidad genética de las progenies, con baja diferenciación genética entre tratamientos. En contraste, las tasas de fecundación, eclosión y mortalidad de peces reproductores revelaron que el SN tuvo la mejor eficiencia reproductiva, un hecho relacionado con el estrés del manejo y las lesiones causadas por el ST. Por lo tanto, el SN demuestra ser el método de desove más adecuado para B. orbignyanus en los programas de repoblación.

Resumo Antecedentes: A manutenção da diversidade genética dos peixes selvagens é uma importante consideração para programas de repovoamento, já que a endogamia pode comprometer a sobrevivência da progênie, além de impactar na resiliência das populações naturais. Objetivo: Avaliar a influência do método de desova (semi-natural - SN ou por extrusão - ST) e número de reprodutores (1♀:3♂ e 2♀:6♂) na eficiência reprodutiva e na diversidade genética de progênie de B. orbignyanus destinados à recuperação de estoques selvagens. Métodos: Taxas de fertilização, eclosão e mortalidade de reprodutores foram monitorados. Para avaliações genéticas (frequências alélicas, heterozigosidade observada e esperada, índice de Shannon, coeficiente de endogamia, análise de variância molecular e diferenciação genética), reprodutores (n=24) e sua progênie (90 larvas/tratamento) foram amostrados e analisados utilizando oito marcadores microssatélites. Resultados: Interação entre método de desova e número de reprodutores não foi significante (p>0,05). Melhores taxas de fertilização e eclosão (p<0,05), e menor (p<0,05) mortalidade de reprodutores foram observados para o método SN, enquanto que o número de reprodutores não afetou esses parâmetros (p>0,05). Os loci microssatélites amplificados produziram um total de 30 alelos, com tamanhos entre 80 e 225 pb e suas frequências indicaram aumento (p<0,05) da diversidade genética nas progênies, mas baixa diferenciação genética entre os tratamentos (p>0,05). Conclusão: Os métodos de desova e números de reprodutores avaliados aumentaram igualmente a diversidade genética das progênies, com baixa diferenciação genética entre tratamentos. Em contraste, as taxas de fecundação, eclosão e mortalidade de reprodutores revelaram que SN obteve a melhor eficiência reprodutiva, fato relacionado com o estresse de manipulação e injurias causadas por ST. Por isso, SN se mostrou como o método de desova mais adequado para B. orbignyanus em programas de repovoamento.

Evaluation of seasonal breeding of the domestic canary (Serinus canaria) in an artificial environment / Avaliação da reprodução sazonal no canário doméstico (Serinus canaria) em ambiente artificial

ABSTRACT: The domestic canary (Serinus canaria) has been bred for hundreds of years to improve the quality of its plumage and its song. Reproduction in this species occurs seasonally, stimulated by a gradual increase in day length. Although, the occurrence of seasonal breeding in canaries is well known, whether canary reproduction can be manipulated remains unknown. Our objective was to determine whether the conditions of captivity (photoperiod and temperature) can be adjusted to enable canaries to reproduce outside of their natural breeding season. Thirty days before the natural breeding season, canary pairs were assigned and separated into three different groups: External Control (housed outdoors under ambient conditions), Artificial Control (housed artificially indoors under conditions similar to the external conditions), and Artificial Altered (housed artificially indoors for five months, with the photoperiod gradually manipulated to simulate that of the natural breeding season) groups. The number of clutches laid was greater in the Artificial Control than in External Control; however, more birds hatched in the External Control. In the Artificial Altered group, the beginning of the breeding season was delayed when the same parameters were used. Although, further research is needed, this study presents new data to assist in the development of protocols that entail gradual changes in environmental conditions to try to reduce the impacts of the processes on animal welfare.

RESUMO: O canário doméstico (Serinus canaria) tem sido criado há centenas de anos para melhorar a qualidade da plumagem e do canto. A reprodução nesta espécie é sazonal, sendo estimulada por um aumento gradual da duração do dia. Embora a reprodução sazonal de canários seja bem conhecida, ainda não se sabe se a reprodução dos canários pode ser manipulada. O objetivo foi determinar se as condições em cativeiro (fotoperíodo e temperatura) podem ser ajustadas para permitir que os canários se reproduzam fora de sua estação natural de reprodução. Trinta dias antes da estação natural de reprodução, os pares foram designados e separados em grupos: Controle Externo (alojado em gaiolas sob condições ambientais externas); Controle Artificial (alojado em gaiolas mantidas em condições artificiais semelhantes ao lado externo); e Alterado Artificial (alojado em gaiolas mantidas em condições artificiais constantes por cinco meses, após o fotoperíodo foi gradualmente manipulado para simular o da estação natural de reprodução). O número de posturas foi maior no grupo Controle Artificial do que no Controle Externo; entretanto, mais aves eclodiram no Controle Externo. No grupo Alterado Artificial, o início do período reprodutivo foi atrasado considerando os mesmos parâmetros reprodutivos. Embora mais pesquisas sejam necessárias, este estudo apresenta novos dados para auxiliar no desenvolvimento de protocolos que promovam mudanças graduais das condições ambientais visando reduzir os impactos no bem-estar animal.

The first larval stage (Zoea I) description of the caridean shrimp Ogyrides occidentalis (Ortmann, 1893) (Alpheoidea: Ogyrididae) reveals congruence with taxonomic status / Descrição do primeiro estágio larval (Zoea I) do camarão carídeo Ogyrides occidentalis (Ortmann, 1893) (Alpheoidea: Ogyrididae) revela congruência com o status taxonômico

Abstract: A complete and detailed description of the first zoeal stage of Ogyrides occidentalis is provided. Larvae were obtained in the laboratory from a female with embryos collected in Ubatuba, State of São Paulo, Brazil. The morphological characters are compared with previous description of the close related O. alphaerostris. Despite of some similarities (number of appendages, pleonites, and setae on the majority of appendages) substantial differences were found between the two species, as the size of larvae and rostrum and segmentation of some structures (antenna exopod, first maxilliped coxa and basis). However, these differences must be interpreted carefully because larval description of O. alphaerostris was conducted before the proposed standardization for decapod larval morphology descriptions. The present larval description furnished additional information to corroborate the recent resurrection of O. occidentalis and will be useful for future comparative and ecological research.

Resumo: Foi realizada uma descrição completa e detalhada da primeira zoea de Ogyrides occidentalis. As larvas foram obtidas em laboratório a partir de uma fêmea com embriões coletada em Ubatuba, estado de São Paulo, Brasil. Os caracteres morfológicos são comparados com a descrição anterior da espécie proximamente relacionada O. alphaerostris. Apesar de algumas semelhanças (número de apêndices, somitos abdominais e cerdas na maioria dos apêndices) foram encontradas diferenças substanciais entre as duas espécies, como o tamanho das larvas e do rostro, e segmentação de algumas estruturas (exopodito da antena e entre coxa e base do primeiro maxilípede). No entanto, essas diferenças devem ser interpretadas com cuidado porque a descrição larval de O. alphaerostris foi realizada antes da padronização proposta para as descrições da morfologia larval de decápodes. A presente descrição larval forneceu informações adicionais para corroborar a recente ressurreição de O. occidentalis e será útil para futuros estudos ecológicos e comparativos.

Long-term monitoring of marine turtle nests in northeastern Brazil / Monitoramento de longo prazo de ninhos de tartarugas marinhas no nordeste do Brasil

Abstract: This study monitored marine turtle nests in a region known as the Potiguar Basin, which stretches from the northern region of Rio Grande do Norte State (5°4'1.15" S, 36°4'36.41" W) to eastern Ceará State (4°38'48.28" S, 37°32'52.08" W) in Brazil. We collected data from January 2011 to December 2019 to identify species of sea turtles that spawn in the basin, to analyze the nesting spatial-temporal pattern and nests characteristics, and to record effects of environmental and anthropic factors on nests. A field team examined sea turtle tracks and nests signs. Turtle clutches were monitored daily until hatchings emerged from the nests. We monitored nests of hawksbill (Eretmochelys imbricata; n = 238) and olive Ridley turtles (Lepidochelys olivacea; n = 103). The nesting season for E. imbricata occurred between December and May and for L. olivacea from March to August. Hawksbills had clutch size, incubation time, number of unhatched eggs, and dead hatchlings higher than olive Ridley turtles; nevertheless, they presented lower hatching success. Precipitation between 0 and 22 mm and relative humidity (RH) higher than 69% increased the hatching success rate for E. imbricata; however, rainfall above 11 mm and RH 64% had the same effect for L. olivacea. Signs of egg theft and human presence (e.g. vehicle traffic and plastic residues on the beach) were recorded and are considered threats to nests. The results of our long-term monitoring study in the Potiguar Basin provide basis for the implementation of mitigation measures and adoption of management policies at nesting beaches in this Brazilian region.

Resumo: Este estudo permitiu o monitoramento de ninhos de tartarugas marinhas em uma região conhecida como Bacia Potiguar, que se estende da região norte do Estado do Rio Grande do Norte (5 ° 4'1,15 "S, 36 ° 4'36,41" W) até o leste do Estado do Ceará (4 ° 38'48,28 "S, 37 ° 32'52,08" W), no Brasil. Coletamos dados de janeiro de 2011 a dezembro de 2019 com o objetivo de identificar as espécies de tartarugas marinhas que desovam na bacia, analisar o padrão espaço-temporal de nidificação, as características dos ninhos, e registrar os efeitos de fatores ambientais e antrópicos. Uma equipe de campo examinou rastros de tartarugas marinhas e sinais de ninhos. As ninhadas das tartarugas foram monitoradas diariamente até que os filhotes emergissem dos ninhos. Monitoramos ninhos de tartarugas-de-pente (Eretmochelys imbricata; n = 238) e tartarugas-oliva (Lepidochelys olivacea; n = 103). A época de nidificação para E. imbricata ocorreu entre dezembro e maio e para L. olivacea de março a agosto. Tartarugas-de-pente apresentaram maior tamanho das ninhadas, tempo de incubação, número de ovos não eclodidos e número de filhotes mortos quando comparado com tartarugas-oliva; no entanto, apresentaram menor sucesso de eclosão. Precipitação entre 0 e 22 mm e umidade relativa (UR) maior que 69% aumentaram a taxa de sucesso de eclosão para E. imbricata; entretanto, chuvas acima de 11 mm e UR 64% tiveram o mesmo efeito para L. olivacea. Sinais de roubo de ovos e presença humana (por exemplo, tráfego de veículos e resíduos de plástico na praia) foram registrados e são considerados ameaças aos ninhos. Os resultados do nosso estudo de monitoramento de longo prazo na Bacia Potiguar fornecem base para a implementação de medidas de mitigação e adoção de políticas públicas em praias de desova nesta região brasileira.

Reproducción en cautiverio de vizcaína Curimata mivartii con extracto pituitario de carpa / Captivity reproduction of vizcaina Curimata mivartii induced with extract of carp pituitary / Reprodução em cativeiro de vizcaína Curimata mivartii com extrato de hipófise de carpa

Resumen Con el objetivo de reproducir vizcaína Curimata mivartii, ejemplares de dos años de edad (n= 54), mantenidos en estanques en tierra (0,1 pez/m2) fueron inducidos con extracto pituitario de carpa (EPC) (Argent, USA) a razón de 6 mg/Kg de peso para las hembras en dos aplicaciones (10 % y 90% con 12 horas entre aplicación); mientras que los machos fueron inducidos con 4,8 mg/Kg EPC en dosis única. Antes de la inducción hormonal a cada hembra le fue realizada biopsia para verificar su estado de maduración gonadal. La muestra de ovocitos fue tratada con solución Serra (Chemí, Colombia) y después de tres minutos, con la ayuda de un estereoscopio de luz, (Leica, EZ4 W, Alemania) en objetivo de 4x, se determinó posición de la vesícula germinal (céntrico, migrando, periférico, sin núcleo). El análisis de la biopsia ovárica mostró que las hembras se encontraban en maduración final. Los machos inducidos se encontraron en fase de espermiación; los cuales a ligera presión abdominal liberaron líquido seminal. Se estimó el índice de ovulación, fecundidad (absoluta y relativa), diámetro de los ovocitos y volumen seminal. El porcentaje de fertilidad fue estimado a las cuatro horas post-eclosión (hpe) y el porcentaje de eclosión fue estimado a las 10 hpe. Las hembras ovularon entre 5 y 6 horas (28±1,0°C) e inmediatamente los huevos fueron obtenidos por extrusión, así como el semen. La fecundidad absoluta fue de 178331,6±28773,7 ovocitos/hembra y la relativa 137,7±19,6 g ovocitos/Kg de hembra. El diámetro de los ovocitos recién desovados fue de 687,2±42,8 µm. La fertilidad fue de 90,7±1,4% y la eclosión de 85,3±1,5%. Los resultados permiten concluir que EPC a dosificación de 6mg/Kg de peso es un buen inductor de la reproducción de vizcaína, con alto índice de ovulación y altos porcentajes de fertilidad y eclosión.

Abstract With the aim of reproducing the vizcaina Curimata mivartii, two-year-old specimens (n = 54), kept in ponds on land (0,1 fish/m2) were induced with carp pituitary extract (EPC) (Argent, USA) at a doses of 6 mg/Kg of weight for females in two applications (10% and 90% with 12 hours between applications), while males were induced with 4,8 mg / Kg EPC in a single dose. Before the hormonal induction, each female underwent an ovarian biopsy to verify the oocytes maturation stage. The oocyte sample was treated with Serra solution (Chemi, Colombia) and after three minutes, with the help of a light stereoscope (Leica, EZ4 W, Germany) in 4x objective, the position of the germinal vesicle was determined (central, migrating, peripheral, breakdown). The analysis of the ovarian biopsy showed that the females were in final maturation. Induced males were found in the spermiation phase; which at light abdominal pressure released seminal fluid. The ovulation index, fecundity (absolute and relative), oocyte diameter and seminal volume were estimated. The percentage of fertility was estimated at four hours after hatching (hah); and the hatching percentage was estimated at 10 hah. The females ovulated between 5 and 6 hours (28±1,0°C) and immediately the eggs were obtained by extrusion, as well as the semen. The absolute fecundity was 178331,6±28773,7 oocytes/female and the relative 137,7±19,6 g oocytes/Kg of female. The diameter of the newly spawned oocytes was 687,2±42,8 µm. Fertility was 90,7±1,4% and hatching 85,3±1,5%. The results allow to conclude that EPC at dosage of 6mg/Kg of weight is a good inducer of the vizcaína reproduction, which high ovulation index and high rates of fertility and hatching.

Resumo Com o objetivo de reproduzir vizcaína Curimata mivartii, espécimenes de dois anos de idade (n = 54), mantidos em tanques em terra (0,1 peixes / m2) foram induzidos com extrato de hipófise de carpa (EPC) (Argent, USA) a uma taxa de 6 mg / kg de peso para as fêmeas em duas aplicações (10% e 90% com 12 horas entre aplicações), enquanto os machos foram induzidos com 4,8 mg / kg de EPC em dose única. Antes da indução hormonal para cada fêmea, uma biópsia ovariana foi realizada para verificar o seu estado de maturação gonadal. A amostra de oócitos foi tratada com solução de Serra (Chemi, Colômbia) e após três minutos, com auxílio de um estereoscópio de luz (Leica, EZ4 W, Alemanha) em objetivo 4x, foi determinada a posição da vesícula germinativa (central, migrando, periférico, sem nucleo). A análise da biópsia ovariana mostrou que as fêmeas estavam em maturação final. Os machos induzidos foram encontrados na fase de espermizaҫã; que na pressão abdominal leve liberou fluido seminal. O índice de ovulação, fecundidade (absoluta e relativa), diâmetro do oócito e volume seminal foram estimados. A percentagem de fertilidade foi estimada em quatro horas pós-eclosão (hpe); enquanto a porcentagem de eclosão foi estimada em 10 hp. As fêmeas ovularam entre 5 e 6 horas (28±1,0°C) e imediatamente os ovos foram obtidos por extrusão, assim como o sêmen. A fecundidade absoluta foi 178331,6±28773,7 oócitos / fêmea e a relativa 137,7±19,6 g oócitos / kg de fêmea. O diâmetro dos oócitos recém-gerados foi 687,2±42,8 µm. A fertilidade foi de 90,7±1,4% e incubação de 85,3±1,5%. Os resultados permitem concluir que a EPC na dosagem de 6mg/Kg de peso é um bom indutor de reprodução de vizcaína, com alto índice de ovulação e altas taxas de fertilidade e eclosão.

Efficacy of three sealing methods on hatchability of micro- cracked eggs from broiler breeder hens / Eficacia de tres métodos de sellado de huevos con micro grietas sobre la incubabilidad de gallinas reproductoras de pollos de engorde / Eficácia de três metodos de selagem de microfissuras em ovos na incubadora provenientes de galinhas reprodutoras de frangos de corte

Abstract Background: Shell fragility of hatching eggs can have negative implications on the economic performance of hatcheries. Objective: To determine the efficacy of sealing eggshell micro cracks with either coloured or uncoloured nail varnish, and molten paraffin on hatchability, embryonic mortality (EM) and hatched chick weight (CW). Methods: Eggs (n= 576) with micro-cracks were assigned among four groups (n=144 each group) for a 21 d incubation period. One group was untreated (CE). In the other groups, the micro cracked area of eggshells was sealed with uncolored nail varnish (NV), colored nail varnish (CV), or molten paraffin wax (MP). A positive control group of un-cracked eggs (UE) was also included (n= 144). Results: The eggshell sealant treatments allowed normal conductance related to egg weight loss after 18 d of incubation (11.45%), and chick weights were normal among treatment groups (44.7 g). Hatchability and embryonic mortality in the early and late incubation periods of the NV group was similar to UE (84.02 vs 86.11% for hatchability, 6.95 vs 10.42% for EM on days 1-10, and 2.08 vs 1.39% for EM on days 18-21 respectively; p>0.05). The CV group had lower hatchability than the NV (77.77 vs 84.02% respectively; p<0.05), whereas MP showed similar hatchability compared to the CE group (59.72 vs 72.92%, respectively p>0.05). Conclusions: Application of uncoloured nail varnish on shell micro-cracks improves egg hatchability.

Resumen Antecedentes: la rotura de la cáscara es un problema que afecta a los huevos para incubación, reduciendo la eficiencia económica de la producción. Objetivo: determinar la eficacia del sellado de micro grietas de huevos utilizando parafina fundida, esmalte de uñas transparente, o esmalte de color sobre la incubabilidad, mortalidad embrionaria (EM) y peso de los pollitos (CW). Métodos: los huevos (n= 576) con micro fisuras fueron asignados a uno de cuatro grupos durante un período de incubación de 21 días. Un grupo se mantuvo sin tratar (CE), en los otros grupos se selló el área micro-agrietada de la cascara con esmalte incoloro de uñas (NV), esmalte de color (CV), o parafina fundida (MP). Tambien se incluyó un grupo de huevos no agrietados (UE, n= 144). Resultados: los tres tratamientos aplicados a la cascara permitieron obtener una pérdida de peso normal de los huevos a 18 d (11,45%), asi como un peso normal de los pollitos (44,7 g). El grupo NV presentó incubabilidad y EM similares a UE durante el periodo de incubación (84,02 vs 86,11% incubalidad y 6,95 vs 10,42% EM entre los días 1-10; y 2,08 vs 1,39% EM entre los días 18-21 respectivamente; p>0,05). El grupo CV tuvo incubabilidad inferior a la de NV (77,77 vs 84,02% respectivamente; p<0,05), mientras que el grupo MP mostró incubabilidad similar a la obtenida por el grupo CE (59,72 vs 72,92%, respectivamente p>0,05). Conclusiones: la aplicación de esmalte incoloro de uñas en las micro-grietas de la cascara mejora la incubabilidad de los huevos.

Resumo Antecedentes: a quebra da casca do ovo é um dos problemas mais importantes durante a incubação, reduzindo a eficiência económica da produção. Objetivo: determinar a eficácia da selagem de microfissuras na casca de ovo com verniz de unhas, transparente ou colorido, e parafina fundida na eclosão, mortalidade embrionária (EM), e peso dos pintos ao nascimento (CW). Métodos: ovos (n= 576) apresentando microfissuras foram divididos em quatro grupos (n= 144) e colocados a incubar por um período de 21 dias. Um dos grupos não foi tratado (CE). Os ovos dos restantes grupos foram selados com verniz de unhas transparente (NV), verniz de unhas colorido (CV) ou parafina fundida (MP). Foi ainda incluido um controlo positivo constituído por ovos sem microfissuras (UE) (n= 144). Resultados: em todos os grupos tratados, observou-se uma perda de peso normal após 18 dias de incubação (11,45%) assim como um peso vivo normal dos pintos ao nascimento (44,7 g). Observou-se uma taxa de eclosão e mortalidade embrionária similar dos períodos iniciais e finais de incubação entre os grupos NV e UE (84,02 vs 86,11% para eclodibilidade, 6,95 vs 10,42% para EM entre os dias 1-10, e 2,08 vs 1,39% para EM entre os dias 18-21 respectivamente; p>0,05). O grupo CV evidenciou uma taxa de eclosão menor do que a do grupo NV (77,77 vs 84,02% respectivamente; p<0,05). No entanto, o grupo MP apresentou uma eclodibilidade similar à observada no grupo CE (59,72 vs 72,92%, respectivamente p>0,05). Conclusões: o uso de verniz de unhas transparente é um método apropriado para incrementar a eclodibilidade de ovos com microfissuras.

Establishment and application of quality control system for detection of schistosomiasis in Jiangsu Province II Evaluation of pathogenic detection capacity of county-level personnel / 中国血吸虫病防治杂志

Objective To evaluate the pathogenic detection capacity of county-level organizations in Jiangsu Province,and to improve the field schistosomiasis detection capacity of professional personnel,thus to provide the technical support for the con-struction of quality control system of schistosomiasis field detection.Method The eggs of Schistosoma japonicum were obtained from rabbit schistosomiasis models and were produced into suspensions at four different concentrations.The county-level workers were invited to hatch the eggs,and the accuracy,detection error rate and omission rate were compared between the detection re-sults and the standard results.The single-blind method was used in the capacity examination. Result A total of 560 suspen-sions were detected by 28 counties(districts,cities),and 283 positive samples and 203 negative samples were detected.The total accuracy was 86.79%,total error rate was 9.38%,and total omission rate was 15.77%.The difference between the detection re-sult and standard result was statistically significant(χ2=12.99,P<0.01).Twenty out of 28 counties(districts,cities)had omis-sion detections,accounting for 71.43%;and 13 had fault detections,accounting for 46.43%.The error rates of workers from the river marshland,hilly areas,water networks,and lake marshland ranged from 4.55% to 43.75%,and the difference was statistical-ly significant(χ2=30.34,P<0.01).The omission rate ranged from 4.17% to 20.45%,and the difference was not statistically sig-nificant(χ2=5.09,P=0.17).The error rates and omission rates of workers from the transmission control areas and transmission interruption areas were 7.50%,13.33% and 10.42%,17.13%,respectively,and the differences were not statistically significant (χ2=0.229,0.575,both P>0.05).The error rates and omission rates of workers from the areas with or without at least ten years history of transmission control were 11.81%,5.00% and 16.67%,14.17%,respectively,and the differences were not statistically significant(χ2=2.804,2.848,both P>0.05).The error rates of workers from the areas with or without at least ten years history of transmission interruption were 11.54% and 10.00%, respectively, and the difference was not statistically significant(χ2=0.069,P=0.792),while the correspondent omission rates were 10.90% and 35.00% respectively,and the difference was statisti-cally significant(χ2=17.364,P<0.01).Conclusion The detection error and omission exist in the schistosomiasis examina-tions in the county-level organizations in Jiangsu Province,and therefore,the field pathogen detection capacity of the professional personnel needs to be further improved.

Assisted Hatching in Couples with Advanced Maternal Age: A Systematic Review and Meta-analysis / 华中科技大学学报（医学）（英德文版）

This systematic review aimed to evaluate the efficacy and safety of assisted hatching (AH) performed in couples with advanced maternal age.We searched for randomized controlled trials (RCTs) in electronic databases,including MEDLINE,EMBASE and CENTRAL (from inception to January 2018);in addition,we hand-searched the reference lists of included studies and similar reviews.We included RCTs comparing AH versus no treatment (control).The meta-analysis was performed by RevMan 5.3 software.The search retrieved 943 records and 8 RCTs were included,comprising 870 cycles (n=440 for AH,and n=430 for control).There was no significant difference in the rates of live birth (RR 0.88,95％ CI 0.65 to 1.18,3 RCTs,n=427,I2=0％),clinical pregnancy (RR 1.00,95％ CI 0.83 to 1.19,8 RCTs,n=870,I2=22％),implantation (RR 1.07,95％ CI 0.83 to 1.39,4 RCTs,n=1359,I2=0％),miscarriage (RR 1.13,95％ CI 0.66 to 1.94,2 RCTs,n=116,I2=0％) and multiple pregnancy (RR 0.89,95％ CI 0.31 to 2.52,1 RCT,n=97,I2=not applicable) between the treatment group and control group.No reasonable conclusions could be drawn regarding reproductive outcomes after AH in patients with advanced maternal age due to the small sample pooled in meta-analyses.Studies of high methodological quality and with adequate power are necessary to further investigate the value of AH in assisted conception of those patients.

Assisted Hatching in Couples with Advanced Maternal Age: A Systematic Review and Meta-analysis / 华中科技大学学报（医学）（英德文版）

This systematic review aimed to evaluate the efficacy and safety of assisted hatching (AH) performed in couples with advanced maternal age.We searched for randomized controlled trials (RCTs) in electronic databases,including MEDLINE,EMBASE and CENTRAL (from inception to January 2018);in addition,we hand-searched the reference lists of included studies and similar reviews.We included RCTs comparing AH versus no treatment (control).The meta-analysis was performed by RevMan 5.3 software.The search retrieved 943 records and 8 RCTs were included,comprising 870 cycles (n=440 for AH,and n=430 for control).There was no significant difference in the rates of live birth (RR 0.88,95％ CI 0.65 to 1.18,3 RCTs,n=427,I2=0％),clinical pregnancy (RR 1.00,95％ CI 0.83 to 1.19,8 RCTs,n=870,I2=22％),implantation (RR 1.07,95％ CI 0.83 to 1.39,4 RCTs,n=1359,I2=0％),miscarriage (RR 1.13,95％ CI 0.66 to 1.94,2 RCTs,n=116,I2=0％) and multiple pregnancy (RR 0.89,95％ CI 0.31 to 2.52,1 RCT,n=97,I2=not applicable) between the treatment group and control group.No reasonable conclusions could be drawn regarding reproductive outcomes after AH in patients with advanced maternal age due to the small sample pooled in meta-analyses.Studies of high methodological quality and with adequate power are necessary to further investigate the value of AH in assisted conception of those patients.

Thinning and drilling laser-assisted hatching in thawed embryo transfer: A randomized controlled trial / 대한생식의학회지

OBJECTIVE: In frozen and thawed embryos, the zona pellucida (ZP) can be damaged due to hardening. Laser-assisted hatching (LAH) of embryos can increase the pregnancy rate. This study compared thinning and drilling of the ZP before frozen embryo transfer (FET). METHODS: Patients were randomly allocated into two groups for LAH using thinning or drilling on day 2 after thawing. Twenty-five percent of the ZP circumference and 50% of the ZP thickness was removed in the thinning group, and a hole 40 µm in diameter was made in the drilling group. RESULTS: A total of 171 in vitro fertilization/intracytoplasmic sperm injection FET cycles, including 85 cycles with drilling LAH and 86 cycles with thinning LAH, were carried out. The thinning group had a similar β-human chorionic gonadotropin-positive rate (38.4% vs. 29.4%), implantation rate (16.5% vs. 14.4%), clinical pregnancy rate (36.0% vs. 25.9%), miscarriage rate (5.8% vs. 2.4%), ongoing pregnancy rate (30.2% vs. 23.5%), and multiple pregnancy rate (7.0% vs. 10.6%) to the drilling LAH group. There were no significant differences in pregnancy outcomes between subgroups defined based on age (older or younger than 35 years) or ZP thickness (greater or less than 17 µm) according to the LAH method. CONCLUSION: The present study demonstrated that partial ZP thinning or drilling resulted in similar outcomes in implantation and pregnancy rates using thawed embryos, irrespective of women's age or ZP thickness.

Effects of three-area laser-assisted zona thinning in 8-cell human embryos on pregnancy outcomes in vitro fertilization / 대한생식의학회지

OBJECTIVE: This study conducted a preliminary examination of the effects of three-area laser-assisted zona thinning (LAZT) during the cleavage stage of embryo development on the hatching process in human in vitro fertilization-embryo transfer (IVF-ET) with subjects of advanced female age or frozen-thawed (FT) embryos. METHODS: Eight-cell stage embryos were treated with LAZT in three areas of the zona pellucida at 120° intervals. The control group was embryos without LAZT. Of the 72 consecutive fresh cycles and the 28 FT embryo transfer cycles, the patients in 55 fresh cycles and 17 FT cycles declined LAZT, and those cycles were defined as the control group. RESULTS: In the fresh cycles, the pregnancy rates were similar in the LAZT and control groups. However, in the FT cycles, the pregnancy rate was significantly higher in the LAZT group than in the control group (45.5% in the LAZT group vs. 23.5% in the control group, p < 0.05). CONCLUSION: These results show that multi-area LAZT resulted in significantly improved pregnancy outcomes in human 8-cell embryos compared to controls.

Metaplasia in swim bladder epithelium of the endangered annual fish Austrolebias nigrofasciatus (Cyprinodontiformes: Rivulidae) results in inadequate swimming and delayed growth

Laboratory breeding of annual fish is often impaired when fish fail to inflate the swim bladder after hatching. These fish, which are known as belly-sliders, cannot swim in the water column, moving mainly on the bottom of water. Therefore, we wanted to understand the factors underlying failed inflation of swim bladder in the annual fish Austrolebias nigrofasciatus and determine the implications of this condition on the growth of juveniles. To identify how the initial filling of the gaseous vesicle is compromised, and to determine the implications of this condition on juvenile growth, we analysed the histology of the swim bladder of both normal swimming fish and belly-sliders during their initial growth phase (first month of life). In addition to slower growth rate compared with fish swimming normally, we found that belly-sliders have metaplasia characterized by epithelium peeling of the swim bladder and the presence of haemorrhage. Based on our findings, it can be concluded that metaplasia of the swim bladder is recurrent in annual fish bred in the laboratory, which explains the large number of unviable larvae. However, its causes remain unknown.(AU)

A criação de peixes anuais é frequentemente comprometida pela ocorrência de peixes que não inflam a vesícula gasosa após a eclosão. Estes peixes não têm capacidade de se manter e nadar na coluna d'água e se locomovem no fundo da água, por isso são chamados belly-sliders. Portanto, objetivamos entender os fatores subjacentes à inflação falhada da bexiga natatória no peixe anual Austrolebias nigrofasciatus e determinar as implicações dessa condição no crescimento de juvenis. Para identificar de que forma o enchimento inicial da vesícula gasosa é comprometido, e determinar as implicações desta condição no crescimento dos juvenis, analisamos histologicamente a vesícula gasosa de peixes com natação normal e de belly-sliders durante o crescimento inicial (primeiro mês de vida). Verificamos que os belly-sliders apresentam metaplasia com descamação do epitélio da vesícula gasosa e presença de hemorragia, além de apresentar crescimento mais lento em relação aos peixes com natação normal. Concluímos que a incidência desta patologia é um problema recorrente e de grande relevância para a manutenção de peixes anuais em laboratório, sendo responsável por uma grande quantidade de juvenis inviáveis, no entanto suas causas permanecem desconhecidas.(AU)

Desarrollo embrionario del capitán de la sabana, Eremophilus mutisii (Pisces: Trichomycteridae) / Embryonic development of Eremophilus mutisii, "the captain of the savannah" (Pisces: Trichomycteridae) / Desenvolvimento embrionário do capitán de la sabana, Eremophilus mutisii (Pisces: Trichomycteridae)

Resumen Eremophilus mutisii es una especie nativa, endémica, del altiplano central colombiano, donde tiene un carácter emblemático y con características que permiten inferir algún potencial piscícola. Con el objetivo de avanzar en el conocimiento sobre aspectos relacionados con su reproducción, se adelantó un seguimiento del desarrollo embrionario bajo condiciones controladas de laboratorio. Para ello, ejemplares silvestres y sexualmente maduros fueron inducidos con extracto de hipófisis de carpa, en dosis totales de 5,5 mg kg-1 en hembras y 4,0 mg kg-1 en machos, siguiendo protocolos con respuesta demostrada. La fertilización se realizó en seco y la incubación se adelantó en frascos de vidrio de 3,6 L a 16,15 ± 1,1 °C. Se colectaron muestras cada hora desde ovocélula hasta eclosión, y se establecieron los estadios por medio de observación en estereoscopio y con una descripción también apoyada con material fotográfico. Se trata de huevos esféricos, adherentes, con 1052 ± 60,2 de diámetro, telolecíticos, y con segmentación meroblástica. Para los estadios de cigoto, clivajes, blastomeración, blástula, gástrula, organogénesis y eclosión, la descripción se realiza en 20 momentos clave, referenciados temporalmente en horas posfertilización (HPF), grados hora (grados h-1) y grados día (grados d-1). Comparado con otros bagres nativos, el periodo de desarrollo se considera extenso, con eclosión completa a las 100 HPF (1615 grados h-1; 387,6 grados d-1). Las larvas nacen con una longitud total de 3148 ± 176 m, con un saco vitelino de forma redondeada con un diámetro medio de 0,9 ± 0,08 mm y un volumen de 0,382 mm3.

Abstract Eremophilus mutisii is a native, endemic species of the Colombian central highlands, with an emblematic character and characteristics that allow inferring some fish farming potential. In order to advance our knowledge about aspects related to its reproduction, embryonic development was monitored under controlled laboratory conditions. To this effect, wild and sexually mature specimens were induced with carp pituitary extract, in total doses of 5.5 mg kg-1 in females and 4.0 mg kg-1 in males, following protocols with demonstrated response. Fertilization was performed dry and incubation was carried out in glass vials of 3.6 L at 16.15 ± 1.1 °C. Samples were collected every hour from egg cells to hatching, and stages were established by means of stereoscopic observation and described with supporting photographic material. They are spherical, adherent eggs, with a diameter of 1052 ± 60.2 pn, telolecithal, and with meroblastic segmentation. For the stages of zygote, cleavages, blastocyst formation, blastula, gastrula, organogenesis, and hatching, description was done in 20 key moments, referenced temporarily in post-fertilization hours (PFH), degrees/hour (degrees h-1), and degrees/day (degrees d-1). Compared with other native catfish, the development period is considered extensive, with full hatching at 100 PFH (1615 degrees h-1; 387.6 degrees d-1). The larvae were born with a total length of 3148 ± 176 m, with a rounded yolk sac with a mean diameter of 0.9 ± 0.08 mm and a volume of 0.382 mm3.

Resumo Eremophilus mutisii é uma espécie nativa, endêmica, do altiplano central colombiano, onde têm um caráter emblemático e com características que permitem inferir algum potencial piscícola. Com o objetivo de avançar no conhecimento sobre aspectos relacionados com sua reprodução, fez-se um seguimento do desenvolvimento embrionário sob condições controladas de laboratório. Para isso, exemplares silvestres e sexualmente maduros foram induzidos com extrato de hipófise de carpa, em doses totais de 5,5 mg Kg-1 em Fêmeas e 4,0 mg Kg-1 em machos, seguindo protocolos com resposta demonstrada. A fertilização realizou-se em seco e a incubação foi realizada em frascos de vidro de 3,6 L a 16,15 ± 1,1 °C. coletaram-se amostras cada hora desde ovocélula hasta eclosão, e se estabeleceram os estados por meio de observação em estereoscópio e com uma descrição também apoiada com material fotográfico. Se trata de ovos esféricos, aderentes, com 1052 ± 60,2 de diâmetro, telolécitos, e com segmentação meroblástica. Para os estados de zigoto, clivagens, blastomeração, blástula, gástrula, organogênese e eclosão, a descrição se realiza em 20 momentos chave, referenciados temporariamente em horas pós-fertilização (HPF), graus hora (graus h-1) e graus dia (graus d-1). Comparado com outros bagres nativos, o período de desenvolvimento se considera extenso, com eclosão completa às 100 HPF (1615 graus h-1; 387,6 grados d-1). As larvas nascem com uma longitude total de 3148 ± 176 m, com um saco vitelino de forma arredondada com um diâmetro médio de 0,9 ± 0,08 mm e um volume de 0,382 mm3.

Eggs viability of Aedes aegypti Linnaeus (Diptera, Culicidae) under different environmental and storage conditions in Manaus, Amazonas, Brazil / Viabilidade de ovos de Aedes aegypti (Diptera, Culicidae) em diferentes condições de armazenamento em Manaus, Amazônia, Brasil

Abstract The viability of Aedes aegypti eggs was assessed in the Amazon region. The eggs were maintained under different conditions: indoors (insectarium) and outdoors (natural environment), as well as in different storage types (plastic cup, paper envelope, plastic bag) for different days. Egg viability was measured as the mean of hatchings observed from egg-bearing sheets of filter paper immersed in water, using three sheets randomly selected from each storage type and at both sites. There were significant differences in the viability of Ae. aegypti eggs with respect to the location (F=30.40; DF=1; P<0.0001), storage type (F=17.66; DF=2; P<0.0001), and time of storage (F=49.56; DF=9; P<0.0001). The interaction between storage site versus storage type was also significant (F=15.96; DF=2; P<0.0001). A higher hatching mean was observed for the eggs kept in the insectarium than for those outdoors (32.38 versus 7.46). Hatching rates of egg batches stored for 12 to 61 days ranged between 84 and 90%. A reduction was observed between 89 and 118 days, with values of 63 and 48%, respectively. With respect to type of storage, mean egg hatching was higher for the eggs in plastic cups (44.46). It was concluded that the viability of the eggs of Ae. aegypti in the Amazon region remains high up to 4 months, after which it declines drastically, although in this study hatching occurred for up to 8 months in very low percentages.

Resumo Testou-se a viabilidade dos ovos de Ae. aegypti na região amazônica armazenados em copos plásticos, envelopes de papel e sacos plásticos que foram mantidos em área interna e em área externa por dias diferentes. Verificaram-se diferenças significativas na viabilidade dos ovos considerando-se a localização (F=30,40; GL=1; P<0,0001), os tipos de armazenamento (F=17,66; GL=2; P<0,0001) e os tempos (F=49,56; GL=9; P<0,0001) e na interação localização versus tipo de armazenamento (F=15,96; GL=2; P<0,0001). A média de eclosão dos ovos armazenados no insetário foi maior: 32,38, do que a dos lotes armazenados na área externa: 7,46. As taxas de eclosão dos ovos armazenados de 12 a 61 dias ficaram entre 90% e 84% com reduções a partir de 89 e 118 dias, quando atingiu de 63% e 48%, respectivamente. Em relação aos tipos de armazenamento a média de ovos dos copos foi significativa (44,46). Conclui-se que a viabilidade dos ovos de Ae. aegypti na região amazônica é mantida em níveis elevados até 4 meses, e a partir daí ocorrem reduções drásticas, com eclosões até 8 meses em percentuais muito baixos.

Effects of laser-assisted hatching and exposure time to vitrification solution on mouse embryo development / 대한생식의학회지

OBJECTIVE: This study was conducted to investigate the efficacy of laser-assisted hatching (LAH) and various vitrification times for embryonic development and blastocyst cell numbers. METHODS: First, 2-cell and 8-cell embryos were collected by flushing out the oviducts. In the control groups, they were vitrified for 8 or 10 minutes without LAH. The LAH groups underwent quarter laser zona thinning-assisted hatching before vitrification (4, 6, and 8 minutes or 4, 7, and 10 minutes, respectively). After incubation, double-immunofluorescence staining was performed. RESULTS: The hatched blastocyst rate 72 hours after the 2-cell embryos were thawed was significantly higher in the 2LAH-ES8 group (33.3%) than in the other groups (p < 0.05). In the control-8 group (22.1±4.6), the cell number of the inner cell mass was higher than in the LAH groups (p < 0.05). The number of trophectoderm cells was higher in the 2LAH-ES6 group (92.8±8.9) than in the others (p < 0.05). The hatched blastocyst rate 48 hours after the 8-cell embryos were thawed was higher in the 8LAH-ES4 group (45.5%) than in the other groups, but not significantly. The inner cell mass cell number was highest in the 8LAH-ES7 group (19.5±5.1, p < 0.05). The number of trophectoderm cells was higher in the 8LAH-ES10 group (73.2±12.1) than in the other groups, but without statistical significance. CONCLUSION: When LAH was performed, 2-cell embryos with large blastomeres had a lower hatched blastocyst rate when the exposure to vitrification solution was shorter. Conversely, 8-cell embryos with small blastomere had a higher hatched blastocyst rate when the exposure to vitrification solution was shorter.