Identification of Sex and Female’s Reproductive Stage in Commercial Fish Species through the Quantification of Ribosomal Transcripts in Gonads

The estimation of maturity and sex of fish stocks in European waters is a requirement of the EU Data Collection Framework as part of the policy to improve fisheries management. On the other hand, research on fish biology is increasingly focused in molecular approaches, researchers needing correct identification of fish sex and reproductive stage without necessarily having in house the histological know-how necessary for the task. Taking advantage of the differential gene transcription occurring during fish sex differentiation and gametogenesis, the utility of 5S ribosomal RNA (5S rRNA) and General transcription factor IIIA (gtf3a) in the molecular identification of sex and gametogenic stage was tested in different economically-relevant fish species from the Bay of Biscay. Gonads of 9 fish species (, Atlantic, Atlantic-chub and horse mackerel, blue whiting, bogue, European anchovy, hake and pilchard and megrim), collected from local commercial fishing vessels were histologically sexed and 5S and 18S rRNA concentrations were quantified by capillary electrophoresis to calculate a 5S/18S rRNA index. Degenerate primers permitted cloning and sequencing of gtf3a fragments in 7 of the studied species. 5S rRNA and gtf3a transcript levels, together with 5S/18S rRNA index, distinguished clearly ovaries from testis in all of the studied species. The values were always higher in females than in males. 5S/18S rRNA index values in females were always highest when fish were captured in early phases of ovary development whilst, in later vitellogenic stages, the values decreased significantly. In megrim and European anchovy, where gonads in different oogenesis stages were obtained, the 5S/18S rRNA index identified clearly gametogenic stage. This approach, to the sexing and the quantitative non-subjective identification of the maturity stage of female fish, could have multiple applications in the study of fish stock dynamics, fish reproduction and fecundity and fish biology in general.

The reproductive cycle of the thorny skate (Amblyraja radiata) in the western Gulf of Maine

The thorny skate (Amblyraja radiata) is a large species of skate that is endemic to the waters of the western north Atlantic in the Gulf of Maine. Because the biomass of thorny skates has recently declined below threshold levels mandated by the Sustainable Fisheries Act, commercial harvests from this region are prohibited. We have undertaken a comprehensive study to gain insight into the life history of this skate. The present study describes and characterizes the reproductive cycle of female and male thorny skates, based on monthly samples taken off the coast of New Hampshire, from May 2001 to May 2003. Gonadosomatic index (GSI), shell gland weight, follicle size, and egg case formation, were assessed for 48 female skates. In general, these reproductive parameters remained relatively constant throughout most of the year. However, transient but significant increases in shell gland weight and GSI were obser ved during certain months. Within the cohort of specimens sampled monthly throughout the year, a subset of females always had large preovulatory follicles present in their ovaries. With the exception of June and September specimens, egg cases undergoing various stages of development were observed in the uteri of specimens captured during all other months of the year. For males (n=48), histological stages III through VI (SIII−SVI) of spermatogenesis, GSI, and hepatosomatic index (HSI) were examined. Although there appeared to be monthly fluctuations in spermatogenesis, GSI, and HSI, no significant differences were found. The production and maintenance of mature spermatocysts (SVI) within the testes was observed throughout the year. These findings collectively indicate that the thorny skate is reproductively active year round.

Reproductive biology of carpenter seabream (Argyrozona argyrozona) (Pisces: Sparidae) in a marine protected area

The carpenter seabream (Argyrozona argyrozona) is an endemic South African sparid that comprises an important part of the handline fishery. A three-year study (1998−2000) into its reproductive biology within the Tsitsikamma National Park revealed that these fishes are serial spawning late gonochorists. The size at 50% maturity (L50) was estimated at 292 and 297 mm FL for both females and males, respectively. A likelihood ratio test revealed that there was no significant difference between male and female L50 (P>0.5). Both monthly gonadosomatic indices and macroscopically determined ovarian stages strongly indicate that A. argyrozona within the Tsitsikamma National Park spawn in the astral summer between November and April. The presence of postovulatory follicles (POFs) confirmed a six-month spawning season, and monthly proportions of early (0−6 hour old) POFs showed that spawning frequency was highest (once every 1−2 days) from December to March. Although spawning season was more highly correlated to photoperiod (r = 0.859) than temperature (r = −0.161), the daily proportion of spawning fish was strongly correlated (r= 0.93) to ambient temperature over the range 9−22oC. These results indicate that short-term upwelling events, a strong feature in the Tsitsikamma National Park during summer, may negatively affect carpenter fecundity. Both spawning frequency and duration (i.e., length of spawning season) increased with fish length. As a result of the allometric relationship between annual fecundity and fish mass a 3-kg fish was calculated to produce fivefold more eggs per kilogram of body weight than a fish of 1 kg. In addition to producing more eggs per unit of weight each year, larger fish also produce significantly larger eggs.

Variability in spawning frequency and reproductive development of the narrow-barred Spanish mackerel (Scomberomorus commerson) along the west coast of Australia

The narrow-barred Spanish mackerel (Scomberomorus commerson) is widespread throughout the Indo-West Pacific region. This study describes the reproductive biology of S. commerson along the west coast of Australia, where it is targeted for food consumption and sports fishing. Development of testes occurred at a smaller body size than for ovaries, and more than 90% of males were sexually mature by the minimum legal length of 900 mm TL compared to 50% of females. Females dominated overall catches although sex ratios within daily catches vary considerably and females were rarely caught when spaw n ing. Scomberomorus commerson are seasonally abundant in coastal waters and most of the commercial catch is taken prior to the reproductive season. Spawning occurs between about August and November in the Kimberley region and between October and January in the Pilbara region. No spawning activity was recorded in the more southerly West Coast region, and only in the north Kimberley region were large numbers of fish with spawning gonads collected. Catches dropped to a minimum when spawning began in the Pilbara region, when fish became less abundant in inshore waters and inclement weather conditions limited fishing on still productive offshore reefs. Final maturation and ovulation of oocytes took place within a 24-hour period, and females spawned in the afternoon-evening every three days. A third of these spawning females released batches of eggs on consecutive days. Relationships between length, weight, and batch fecundity are presented.

A riqueza e diversidade de anfíbios anuros da Serra do Mendanha, Estado do Rio de Janeiro, RJ: grau de conservação da floresta, variação altitudinal e uso de recursos hídricos

Os anfíbios anuros que habitam o bioma Mata Atlântica, especialmente em sua floresta ombrófila densa, são pouco conhecidos sob diversos aspectos de sua ecologia, existindo poucas informações disponíveis na literatura científica. Estes dados estão limitados a poucas localidades, geralmente estudos realizados em fragmentos remanescentes do sudeste brasileiro. A mata bem conservada que recobre os 3300 ha da Serra do Mendanha, apesar de localizada na região metropolitana do Estado do Rio de Janeiro, ainda não tinha sua anurofauna conhecida. No presente estudo, recolhemos informações ecológicas sobre as espécies que habitam a região, especialmente da assembléia encontrada no folhiço que recobre o solo, com o uso de diferentes metodologias: armadilhas de queda; parcelas cercadas; procura visual e auditiva em transecções e encontros ocasionais; assim como dados abióticos do ambiente regional (profundidade do folhiço; níveis de pH; taxa de oxigênio dissolvido; temperaturas do ar e da água; umidade do ar). Exemplares-testemunho foram fixados e depositados na coleção de anfíbios do Museu Nacional, Rio de Janeiro. A anurofauna da Serra do Mendanha é composta por pelo menos 44 espécies que estão distribuídas em 12 famílias, sendo a família Hylidae com o maior número de espécies (50%, n = 22), enquanto que Physalaemus signifer foi a espécie mais abundante (18%, n = 272), sendo habitante do folhiço. A espécie Rhinella ornata contribuiu com a maior biomassa (m = 548 g). Identificamos uma nova espécie de anuro (Brachycephalidae), que também habita o folhiço das cotas altimétricas acima de 700 m. Biogeograficamente a comunidade de anuros da área estudada indicou ser mais similar (68%) com a comunidade da região da Serra da Tiririca e arredores. Comparando-se os três tipo de fisionomias, ou mesohábitats, existentes na Serra do Mendanha, em termos de riqueza e diversidade, a floresta secundária indicou ter os mais elevados índices, seguido pela floresta pouco perturbada e pela monocultura de bananeiras. A assembléia de anuros que habita o folhiço da Serra do Mendanha é composta por nove espécies, que pouco diferiu ao longo de um gradiente altitudinal (0 a 900 m), com o registro das espécies Euparkerella brasiliensis, H. binotatus, Leptodactylus marmoratus e Zachaenus parvulus na maioria das cotas altimétricas. A altitude, a declividade e profundidade do folhiço foram as variáveis abióticas que mais influenciaram na distribuição de abundância e de riqueza de espécies do folhiço. Na estação úmida (setembro a março) a densidade de anuros no folhiço foi de 10,4 ind./100m2, enquanto que na estação seca (abril a agosto) houve uma redução de 34,6% (6,8 ind./100m2). As assembléias de anuros utilizam os recursos hídricos disponíveis na Serra do Mendanha (água da chuva, córregos, fitotelmas, rios e umidade do ar), de diferentes formas, associadas diretamente ao modo reprodutivo de cada espécie. A altitude, a temperatura da água e o pH afetaram a distribuição de espécies de girinos nos diferentes sítios reprodutivos. O modo reprodutivo 1 foi o mais freqüente (n = 18) entre os 14 modos identificados. O estudo de 12 poças (lênticas e lóticas) indicou que estas diferiram consistentemente entre si, seja nas suas dimensões, na composição de suas assembléias de girinos e nos seus componentes abióticos. Diversos predadores de girinos foram encontrados em algumas poças. A assembléia de girinos de cada poça indicou ser moldada pela interação de diferentes fatores ambientais, ecológicos e filogenéticos de cada espécie