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.

Length at maturity in three pelagic sharks (Lamna nasus, Isurus oxyrinchus, and Prionace glauca) from New Zealand

Reproductive data collected from porbeagle, shortfin mako, and blue sharks caught around New Zealand were used to estimate the median length at maturity. Data on clasper development, presence or absence of spermatophores or spermatozeugmata, uterus width, and pregnancy were collected by observers aboard tuna longline vessels. Direct maturity estimates were made for smaller numbers of sharks sampled at recreational fishing competitions. Some data sets were sparse, particularly over the vital maturation length range, but the availability of multiple indicators of maturity made it possible to develop estimates for both sexes of all three species. Porbeagle shark males matured at 140–150 cm fork length and females at about 170–180 cm. New Zealand porbeagles therefore mature at shorter lengths than they do in the North Atlantic Ocean. Shortfin mako males matured at 180–185 cm and females at 275 –285 cm. Blue shark males matured at about 190 –195 cm and females at 170–190 cm; however these estimates were hampered by small sample sizes, difficulty obtaining representative samples from a population segregated by sex and maturity stage, and maturation that occurred over a wide length range. It is not yet clear whether regional differences in median maturity exist for shortfin mako and

Maturity, Pulp Removal and Storage Effects on the Germination of Livistona rotundifolia Seeds

The Livistona rotundifolia species is native to Oceania, and has a high potential for landscaping use and as a pot plant. This work aimed to study the effects of the maturation stage, pulp removal and storage on the germination of L. rotundifolia seeds. The experimental design was entirely randomized in a factorial arrangement 5x2x2 (five storage periods: 15, 30, 45, 60 and 75 days; two maturation stages: green and ripe; and the presence or absence of the pulp - exocarp and mesocarp) with four replications of 25 seeds each. After sorting out the fruits by the maturity stage and removing the pulp out of half of the fruits from each plot, the seeds were placed in closed bottles, which were sealed and stored in a cold chamber at 10 degrees C. The seeds were removed from the cold chamber and left to germinate in plastic boxes (gerbox type) with sphagnum. The boxes were kept at 25-35 degrees C and photoperiod of 12 hours. The germination rate was determined when seed germination was steady. The highest germination rate was found when green fruits had their pulp removed. The germination rate gradually decreased with the increase of the storage period regardless the maturation stage and the presence or absence of the pulp.

Modeling systems and vis/NIR device to improve peach and nectarine pre and post-harvest fruit maturity management

During the last decade peach and nectarine fruit have lost considerable market share, due to increased consumer dissatisfaction with quality at retail markets. This is mainly due to harvesting of too immature fruit and high ripening heterogeneity. The main problem is that the traditional used maturity indexes are not able to objectively detect fruit maturity stage, neither the variability present in the field, leading to a difficult post-harvest management of the product and to high fruit losses. To assess more precisely the fruit ripening other techniques and devices can be used. Recently, a new non-destructive maturity index, based on the vis-NIR technology, the Index of Absorbance Difference (IAD), that correlates with fruit degreening and ethylene production, was introduced and the IAD was used to study peach and nectarine fruit ripening from the “field to the fork”. In order to choose the best techniques to improve fruit quality, a detailed description of the tree structure, of fruit distribution and ripening evolution on the tree was faced. More in details, an architectural model (PlantToon®) was used to design the tree structure and the IAD was applied to characterize the maturity stage of each fruit. Their combined use provided an objective and precise evaluation of the fruit ripening variability, related to different training systems, crop load, fruit exposure and internal temperature. Based on simple field assessment of fruit maturity (as IAD) and growth, a model for an early prediction of harvest date and yield, was developed and validated. The relationship between the non-destructive maturity IAD, and the fruit shelf-life, was also confirmed. Finally the obtained results were validated by consumer test: the fruit sorted in different maturity classes obtained a different consumer acceptance. The improved knowledge, leaded to an innovative management of peach and nectarine fruit, from “field to market”.

1-Methylcyclopropene Delays Softening in Tomato Slices

I-Methylcyclopropene (1-MCP) has the potential in tomato to reduce ethylene-associated changes in texture. Tomato cv. 'Revolution' was harvested at the 'pink' maturity stage and whole fruit treated with 0, 0.1, 1.0 or 10.0 µL.L-' 1-MCP at 20 "C for 12 h. Slices of 7-mm thickness were cut using a commercial slicer, and the slices stored in vertical stacks in plastic containers at 5°C for 7 days. The application of 1-MCP reduced both ethylene production and respiration rate of slices and resulted in firmer pericarp firmness. Ethylene production was 24%, 40%, and 62% lower following 0.1, 1.0, 10.0 µL L-' 1-MCP, respectively, compared with controls. In addition, respiration rate was reduced 6%, 10% and 20% by those 1-MCP treatments. 1-MCP treatments produced 20%, 34%, and 24% higher pericarp firmness, respectively, than in fruit not treated with 1-MCP.

Effect of an Ethylene Absorbent on Quality of Tomato Slices

Ethylene production is stimulated during the slicing of fresh cut tomato slices. Experiments were conducted to investigate whether the inclusion of ethylene absorbents in packaging affects the quality of tomato slices cv. Revolution during storage at 5OC. ‘Pink’ maturity stage tomatoes were cut into 7mm thick slices and vertically stacked in closed glass containers for 12 days with or without Purafil® to remove ethylene. The ethylene removal treatment resulted in reduced ethylene, less CO2 accumulation, and firmer slices.

The Application of Near Infrared Spectroscopy for the Assessment of Avocado Quality Attributes, Infrared Spectroscopy. Chapter 14

Quality and safety evaluation of agricultural products has become an increasingly important consideration in market/commercial viability and systems for such evaluations are now demanded by customers, including distributors and retailers. Unfortunately, most horticultural products struggle with delivering adequate and consistent quality to the consumer. Removing inconsistencies and providing what the consumer expects is a key factor for retaining and expanding both domestic and international markets. Most commercial quality classification systems for fruit and vegetables are based on external features of the product, for example: shape, colour, size, weight and blemishes. However, the external appearance of most fruit is generally not an accurate guide to the internal or eating quality of the fruit. Internal quality of fruit is currently subjectively judged on attributes such as volatiles, firmness, and appearance. Destructive subjective measures such as internal flesh colour, or objective measures such as extraction of juice to measure sweetness (oBrix) or assessment of dry matter (DM) content are also used, although obviously not for every fruit – just a sample to represent the whole consignment. For avocado fruit, external colour is not a maturity characteristic, and its smell is too weak and appears later in its maturity stage (Gaete-Garreton et al., 2005). Since maturity is a major component of avocado quality and palatability, it is important to harvest mature fruit, so as to ensure that fruit will ripen properly and have acceptable eating quality. Currently, commercial avocado maturity estimation is based on destructive assessment of the %DM, and sometimes percent oil, both of which are highly correlated with maturity (Clark et al., 2003; Mizrach & Flitsanov, 1999). Avocados Australia Limited (AAL (2008)) recommend a minimum maturity standard for its growers of 23 %DM (greater than 10% oil content) for the ‘Hass’ cultivar, although consumer studies indicate a preference for at least 25 %DM (Harker et al., 2007).

Fusarium graminearum and Fusarium pseudograminearum caused the 2010 head blight epidemics in Australia

Wheat crops in southeast Queensland (Qld) and northern New South Wales (NSW) were infected with fusarium head blight (FHB)-like symptoms during the 201011 wheat growing season. Wheat crops in this region were surveyed at soft dough or early maturity stage to determine the distribution, severity, aetiology and toxigenicity of FHB. FHB was widespread on bread wheat and durum, and Fusarium graminearum and/or F.pseudograminearum were diagnosed from 42 of the 44 sites using species-specific PCR primers directly on spikelets or from monoconidial cultures obtained from spikelets. Stem base browning due to crown rot (CR) was also evident in some samples from both states. The overall FHB and CR severity was higher for NSW than Qld. Deoxynivalenol (DON) concentration of immature grains was more than 1 mg kg-1 in samples from 11 Qld and 14 NSW sites, but only 13 of 498 mature grain samples sourced from the affected areas had more than 1 mg kg-1 DON. DON concentration in straw also exceeded 1 mg kg-1 in eight Qld and all but one NSW sites but this was not linked to DON concentration of immature grains. The proportion of spikelets with positive diagnosis for F.graminearum and/or F.pseudograminearum and weather-related factors influenced DON levels in immature grains. The average monthly rainfall for AugustNovember during crop anthesis and maturation exceeded the long-term monthly average by 10150%. Weather played a critical role in FHB epidemics for Qld sites but this was not apparent for the NSW sites, as weather was generally favourable at all sites.

Zeaxanthin biofortification of sweet-corn and factors affecting zeaxanthin accumulation and colour change

Zeaxanthin, along with its isomer lutein, are the major carotenoids contributing to the characteristic colour of yellow sweet-corn. From a human health perspective, these two carotenoids are also specifically accumulated in the human macula, and are thought to protect the photoreceptor cells of the eye from blue light oxidative damage and to improve visual acuity. As humans cannot synthesise these compounds, they must be accumulated from dietary components containing zeaxanthin and lutein. In comparison to most dietary sources, yellow sweet-corn (Zea mays var. rugosa) is a particularly good source of zeaxanthin, although the concentration of zeaxanthin is still fairly low in comparison to what is considered a supplementary dose to improve macular pigment concentration (2 mg/person/day). In our present project, we have increased zeaxanthin concentration in sweet-corn kernels from 0.2 to 0.3 mg/100 g FW to greater than 2.0 mg/100 g FW at sweet-corn eating-stage, substantially reducing the amount of corn required to provide the same dosage of zeaxanthin. This was achieved by altering the carotenoid synthesis pathway to more than double total carotenoid synthesis and to redirect carotenoid synthesis towards the beta-arm of the pathway where zeaxanthin is synthesised. This resulted in a proportional increase of zeaxanthin from 22% to 70% of the total carotenoid present. As kernels increase in physiological maturity, carotenoid concentration also significantly increases, mainly due to increased synthesis but also due to a decline in moisture content of the kernels. When fully mature, dried kernels can reach zeaxanthin and carotene concentrations of 8.7 mg/100 g and 2.6 mg/100 g, respectively. Although kernels continue to increase in zeaxanthin when harvested past their normal harvest maturity stage, the texture of these 'over-mature' kernels is tough, making them less appealing for fresh consumption. Increase in zeaxanthin concentration and other orange carotenoids such as p-carotene also results in a decline in kernel hue angle of fresh sweet-corn from approximately 90 (yellow) to as low as 75 (orange-yellow). This enables high-zeaxanthin sweet-corn to be visually-distinguishable from standard yellow sweet-corn, which is predominantly pigmented by lutein.

Reprodução de peixes de riachos: uma abordagem comparativa entre as estratégias r e K

No presente trabalho, objetivou-se caracterizar a estratégia reprodutiva, enfatizando o investimento energético, de duas espécies de peixe do rio Ubatiba, Maricá, Rio de Janeiro: Parotocinclus maculicauda (K-estrategista) e Astyanax hastatus (r-estrategista). Foram realizadas coletas bimestrais de Junho de 2010 a Abril de 2011 totalizando 236 exemplares amostrados de A. hastatus e 234 de P. maculicauda. Para cada exemplar foram registrados os dados de comprimento padrão (Cp, cm), peso total (Pt, g), peso gonadal (Pg, g), sexo e estádio de maturação. Através da estrutura de tamanho, observamos que as fêmeas atingem maior comprimento, em relação aos machos, para as duas espécies. A relação peso/ comprimento evidenciou para ambas as espécies, crescimento alométrico negativo (inferior a 3), demonstrando crescimento mais longelíneo. Para a proporção sexual, o teste χ2 foi aplicado e indicou que, para as duas espécies, há significativamente mais fêmeas. A distribuição sexual no ano mostrou que as fêmeas se mantêm em maioria durante todo o ano para P. maculicauda. Para A. hastatus este padrão também se mantém, porém com exceção do bimestre Novembro/Dezembro, quando o número de machos torna-se um pouco maior. O tamanho de primeira maturação mostrou-se o mesmo para ambas as espécies (2,5 a 3,0 cm). A variação temporal da freqüência de indivíduos reprodutivos e não reprodutivos juntamente com a distribuição temporal dos valores individuais de IGS mostrou que P. maculicauda se reproduz com maior intensidade nas estações chuvosas (Setembro a Abril), reduzindo sua atividade reprodutiva de maneira significativa nas estações secas (Maio a Agosto). Já A. hastatus demonstrou regular atividade reprodutiva durante todo o ano com pequeno pico no bimestre Novembro/ Dezembro. Desova do tipo total foi registrada para Astyanax, enquanto que para Parotocinclus registrou-se desova parcelada em três lotes. Em ambas as espécies foi observada relação inversa entre volume e a quantidade de ovócitos produzidos, com A. hastatus produzindo muitos ovócitos (fecundidade: 463 + 213 ovócitos/grama de peixe) de reduzido volume (diâmetro = 800 μm e volume = 0,26 mm3) e P. maculicauda produzindo número bem inferior (fecundidade: 47 + 13 ovócitos/grama de peixe), porém com volume superior (diâmetro = 1.600 μm e volume = 2,14 mm3). Com isso o valor energético relativo também se mostrou superior, com A. hastaus produzindo ovócitos vitelogênicos com 0,4+ 0,08 cal/unidade e P. maculicauda produzindo os mesmos ovócitos com 1,8+ 1,1 cal/unidade. Para a produção energética total investida na produção de gametas, foi considerado o tipo de desova de cada espécie, com Astyanax investindo 50,5 + 24 calorias/grama/grama de peixe e Parotocinclus investindo 88,4 + 72,46 cal/grama/grama de peixe, porém sem diferenças significativas (Mann-Whitney; U =235,0, p=0,08), indicando, portanto que independente da estratégia adotada (r ou K), o gasto energético na produção de ovócitos é a mesma.

Reproductive biology, spawning season, and growth of female rex sole (Glyptocephalus zachirus) in the Gulf of Alaska

Rex sole (Glyptocephalus zachirus) have a wide distribution throughout the North Pacific, ranging from central Baja California to the western Bering Sea. Although rex sole are an important species in the commercial trawl fisheries off the U.S. West Coast, knowledge of their reproductive biology is limited to one study off the Oregon coast where ovaries were analyzed with gross anatomical methods. This study was initiated to determine reproductive and growth parameters specific to rex sole in the Gulf of Alaska (GOA) stock. Female rex sole (n=594) ranging in total length from 166 to 552 mm were collected opportunistically around Kodiak Island, Alaska, from February 2000 to October 2001. All ovaries were analyzed by using standard histological criteria to determine the maturity stage. Year-round sampling of rex sole ovaries confirmed that rex sole are batch spawners and have a protracted spawning season in the GOA that lasts at least eight months, from October to May; the duration of the spawning season and the months of spawning activity are different from those previously estimated. Female rex sole in the GOA had an estimated length at 50% maturity (ML50) of 352 mm, which is greater than the previously estimated ML50 at southern latitudes. The maximum age of collected female rex sole was 29 years, and the estimated age at 50% maturity (MA50) in the GOA was 5.1 years. The von Bertalanffy growth model for rex sole in the GOA was significantly different from the previously estimated model for rex sole off the Oregon coast. This study indicated that there are higher growth rates for rex sole in the GOA than off the Oregon coast and that there are differences in length at maturity and similarity in age at maturity between the two regions.

Condition factor and length-weight relationship of Nemipterus japonicus (Bloch) off Bombay coast

Fluctuations in the K values of Nemipterus japonicus (Bloch) off Bombay coast were interpreted regarding sex, month and females maturity stage. These indicate differential growth rates in males and females. Males and females attain first maturity at 145 mm and 115 mm respectively, second maturity is attained by both the sexes at 195 mm. First spawning occurs when both are of 155 mm length and at second spawning males and females attain 215 and 205 mm of length respectively. The fish mature and breed at "O" year; the main spawning period is from August to November with peak spawning activities in October. It grows about 155 mm in first year at 12.91mm per month and about 215 mm in the second year at 5.0 mm per month on an average. Length-weight relationships for males and females are given. The rate of growth of females by weight was found to be slower below 150 mm, but faster than that of males above 150 mm specimens.

Escala de madurez gonadal de merluza peruana Merluccius gayi peruanus (Ginsburg, 1954)

Se determinó la escala de madurez gonadal macroscópica, validada microscópicamente del recurso merluza peruana Merluccius gayi peruanus. Para ello, se analizaron histológicamente gónadas de hembras y machos colectadas en los cruceros de Evaluación de Recursos Demersales, desde el otoño del 2002 hasta el verano del 2004. En base a las observaciones microscópicas de cada estadio de madurez gonadal, se establecen las características visuales más conspicuas que diferencian a cada uno de ellos, estableciéndose seis estadios tanto para hembras como para machos: 0 (virginal), I (reposo), II (en maduración), III (maduro), IV (desovante/expulsante), V (recuperación/ post expulsante).

Escala de madurez gonadal de cabrilla Paralabrax humeralis (Valenciennes, 1828)

Se realiza la validación de la escala de madurez gonadal macroscópica de la cabrilla Paralabrax humeralis. Se analizaron 197 ovarios y 188 testículos provenientes del seguimiento de la pesquería demersal y cruceros de investigación de recursos demersales en el 2005 (Enero-febrero y mayo-junio). Sobre la base del análisis histológico se asignó a cada individuo un estadio de madurez macroscópico. Se realizó la descripción de los estadios de madurez que van desde el estadio 0 (virginal) hasta el 5 (recuperación en hembras, post expulsante en machos). Se realizó una comparación con escalas anteriormente realizadas, discutiendo además los beneficios de su uso.

Compactação do solo e produção de cultivares de milho em latossolo vermelho: I - características de planta, solo e índice S

Este trabalho teve como objetivo avaliar, em condições de campo, a resposta de duas cultivares de milho com características genéticas distintas em Latossolo Vermelho em seis níveis de compactação. O experimento foi conduzido em faixas, no delineamento de blocos completos casualizados, com quatro repetições. Utilizaram-se os híbridos de milho DKB 390 e DAS 2B710. Após a semeadura do milho, coletaram-se amostras indeformadas de solo nas profundidades de 0-10, 10-20 e 20-30 cm, para determinação de propriedades físicas do solo e índice S. No estádio de maturidade fisiológica dos grãos do milho, foram determinados: a altura das plantas, a altura de inserção da primeira espiga, o diâmetro do segundo colmo acima do solo e a massa de matéria seca das plantas. As espigas foram colhidas e debulhadas para determinação da produtividade de grãos, corrigida para a umidade-padrão de 13 %, calculando-se também o número de espigas por planta e o índice de colheita. O índice S apresentou correlação positiva com a produtividade, porém, abaixo do limite de S < 0,035, estabelecido para uma condição de solo desestruturado, ocorreram perdas acentuadas na produtividade de milho. Os híbridos simples de milho DKB 390 e DAS 2B710 não apresentaram diferenças quando submetidos aos diferentes níveis de compactação. A produtividade dos híbridos de milho foi significativamente menor quando a resistência do solo à penetração atingiu 2,15 MPa.