Phytochemical screening and antibacterial assessment of two macroalgae Ulva papenfussi and Ulva nematoidea (Chlorophyta) against the bacterium Vibrio parahaemolyticus.

This study was conducted to determine whether the macroalgae Ulva papenfussi and Ulva nematoidea could be alternatives for preventing Litopenaeus vannamei vibriosis caused by the bacterium Vibrio parahaemolyticus. Phytochemical screening was performed on methanolic extracts to qualitatively determine the main groups of bioactive compounds, previous to an in vitro antibacterial test against V. parahaemolitycus. Phenols, polyphenols, flavonoids, and the high presence of carbohydrates were found in both macroalgae. U. papenfussi showed more presence of lipids and alkaloids than U. nematoidea . Macroalgae extracts prepared (v:v) with a 1:1 methanol: dichloromethane solvent was used for the in vitro test using the disc diffusion method (MDD). Filter paper discs impregnated with 1.0, 1.5, 2.0, 3.0, and 4.0 mg of the extracts showed antibacterial activity against V. Parahaemolitycus in a dose-dependent manner in both macroalgae. The inhibition zone varied significantly (p < 0.05) from 8.33 ± 0.12 to 11.41 ± 0.73 mm for 1 to 3 mg of extract levels, respectively. In conclusion, both macroalgae have antibacterial activity in their crude extracts against this bacteria. It is suggested to evaluate it as a feed additive for L. vannamei. This study is the first report on a phytochemical screening and antibacterial activity of these macroalgae against V. parahaemolyticus.

Sperm handling in aquatic animals for artificial reproduction.

Artificial reproduction involves collection and handling of gametes in a way that secures their quality and maximizes the fertilization outcome. In addition to initial sperm quality, numerous steps can affect the final result of fertilization, from the sperm collection process until gamete mixing (or co-incubation) when the spermatozoon enters or fuses with the oocyte. In this review, we summarize the whole process of sperm handling, from collection until fertilization for fish, penaeid shrimp, bivalve mollusks and marine mammals. To obtain sperm from captive animals, techniques vary widely across taxa, and include stripping by abdominal massage or testis surgical removal in fish, spermatophore collection in penaeid shrimps, gonadal scarification or temperature shock in bivalve mollusks, and voluntary collection via positive reinforcement in mammals. In most cases, special care is needed to avoid contamination by mucus, seawater, urine, or feces that can either activate sperm motility and/or decrease its quality. We also review techniques and extender solutions used for refrigerated storage of sperm across the aforementioned taxa. Finally, we give an overview of the different protocols for in vivo and in vitro fertilization including activation of sperm motility and methods for gamete co-incubation. The present study provides valuable information regarding breeder management either for animal production or species conservation.

Effects of dietary Bacillus subtilis and Shewanella algae in expression profile of immune-related genes from hemolymph of Litopenaeus vannamei challenged with Vibrio parahaemolyticus.

B. subtilis and S. algae effects in growth, survival and innate immunity were assessed on L. vannamei juveniles. During 60 days, shrimp were reared in three treatments: Bs, fed with 106 CFU of B. subtilis per gram of commercial feed, Sa, fed with 106 CFU of S. algae per gram of commercial feed and Control (without bacterial addition). Then, the animals were subjected to a V. parahaemolyticus challenge. For this purpose, four treatments were established: Control (shrimp not submitted to probiotic treatments), Vibrio (Vibrio challenged shrimp), Vibrio + Bs (Bs challenged shrimp) and Vibrio + Sa (Sa challenged shrimp). Shrimp hemolymph was sampled 45-days after rearing and 24 h post-challenge for quantification of prophenoloxidase (proPO), lipopolysaccharide and ß-1,3-glucan-binding protein (LGBP) and hemocyanin (HEM) transcripts by qPCR. Moreover, shrimp final weight and survival were also verified. B. subtilis administration enhanced shrimp growth and improved proPO, LGBP and HEM expression levels before and after challenge. After 60-days of feeding, Sa final weight was higher than the Control, whereas Vibrio + Sa cumulative mortality after 48 h of Vibrio challenge was lower than Vibrio group. These results could be correlated with the proPO and LGBP up regulation in Vibrio + Sa compared to Vibrio group, protecting L. vannamei from the bacterial infection. Together, these results suggest the probiotic potential of B. subtilis e S. algae in the modulation of immune-related genes as a tool to control V. parahaemolyticus infection inside shrimp.

Technical efficiency analysis of marine shrimp farming (Litopenaeus vannamei) in biofloc and conventional systems: A case study in northeastern Brazil.

Shrimp farming has a great ecological, economic, and social importance in northeastern Brazil. Although the conventional farming system is widely used, biofloc technology (BFT) system has been developed to reduce environmental impacts and optimizing the production. Thus,the present study evaluated the technical efficiency of production cycles of Litopenaeus vannamei in conventional and BFT systems.We analyzed 48 production cycles in the conventional system in 2013 and 2014 and 9 cycles in the BFT system in 2014 through data envelopment analysis (DEA). The inputs corresponded to the population density (post-larvae m-2), amount of feed (kg ha-1 cycle-1), labor (man ha-1) and power supply (HP ha-1), whereas the yield (kg ha-1 cycle-1) was considered the output. The results indicated four production cycles (7.0%) technically efficient, three in the conventional system and one in the BFT. By comparing the productive systems, there was significant difference in their mean scores of technical efficiency. There was a greater influence of the management inefficiency on the conventional system, while the production scale reduced the average scores of technical efficiency in the BFT system. These results may help to improve the development of sustainable L. vannamei farming in both systems by reducing waste and increasing profits.

Coliform risk assessment through use of the clam Anomalocardia brasiliana as animal sentinel for shellfish harvesting areas in Brazil's northeast.

Outbreaks of food-borne diseases related to consumption of contaminated shellfish have been reported in many countries, but not in Brazil, possibly due to deficient reporting. Here we investigated the suitability of the clam Anomalocardia brasiliana as an animal sentinel for coliform monitoring in shellfish harvesting areas of Brazil's northeast. Samples of shellfish meats (40 clams per sample; n = 8 per collection) were collected at random from April 2009 through March 2010 in the bay area of Mangue Seco (state of Pernambuco). The numbers of thermotolerant coliforms were analyzed through the most probable number technique, and these contamination levels were tentatively correlated with the precipitation recorded on the day of sampling or 24 to 48 h beforehand. A. brasiliana shellfish meats from local retail shops (250 g per sample/ n = 3 per market) sold frozen were also investigated from August 2010 through June 2011. We found that the highest coliform contamination levels were correlated with recent rainfall events, limited to 24 h before sampling. However, irrespective of the rainfall level, the mean contamination above the Brazilian legal threshold of < 3 × 10(2) MPN/ 100 g for shellfish harvesting areas ranged from 18.7 to 93.7 % of samples analyzed monthly. Additionally, a large number of samples obtained from retail shops were also highly contaminated by coliforms during rainy periods, and therefore were not proper for human consumption. We conclude that A. brasiliana can be successfully used to monitor the contamination levels of coliforms in shellfish harvesting areas in Brazil's northeast coast.

Análise comparativa da criação dos camarões-rosa Farfantepenaeus brasiliensis e FarfantePenaeus paulensis criados em gaiolas em ambiente estuarino / Comparative analysis of pink shrimp Farfantepenaeus brasiliensis and FarfantePenaeus paulensis reared in estuarine cage culture system

A análise da criação de Farfantepenaeus brasiliensis e Farfantepenaeus paulensis em gaiolas, na Lagoa dos Patos, Rio Grande do Sul (RS), foi realizada a partir de juvenis produzidos em cativeiro com peso médio inicial de aproximadamente 1,2g. O experimento foi realizado em seis gaiolas (três gaiolas/tratamento), com abertura de malha de 5mm, área de fundo de 4m² durante 65 dias. A distribuição dos indivíduos nos tratamentos (F. brasiliensis e F. paulensis) foi aleatória, mantendo-se a densidade de 20 camarões m-2 nas unidades experimentais. A cada 15 dias foram realizadas biometrias para ajuste da quantidade de ração fornecida e avaliação do crescimento dos camarões. Ao final do experimento, foi realizada a biometria total dos camarões para avaliação da sobrevivência. Não houve diferença significativa entre a sobrevivência de F. brasiliensis (94,17 ± 9,04) e F. paulensis (98,50±0,71). O peso médio final foi significativamente maior para o F. brasiliensis (7,98± 0,94g); porém, não foram observadas diferenças significativas na produção de biomassa (127,81±17,93 e 126,65±1,74g m-2) e conversão alimentar aparente (1,39±0,27 e 1,57±0,09) de F. brasiliensis e F. paulensis, respectivamente. Os resultados indicam que F. brasiliensis apresenta potencial para produção em estruturas alternativas e incentivam que novas pesquisas sejam realizadas para o desenvolvimento de um pacote tecnológico de produção dessas espécies.

The cage culture of Farfantepenaeus brasiliensis and Farfantepenaeus paulensis was analyzed in the Patos Lagoon estuary using juveniles produced in captivity. Mean initial weight of the juveniles was approximately 1.2g. The experiment was conducted in 6 cages (3 per treatment), with mesh size of 5mm and bottom area of 4m², during 65 days. The individuals were randomly distributed into two treatments (F. brasiliensis e F. paulensis), keeping the stocking density of 20 shrimps m-2 in the experimental units (cages). Each 15 days shrimps were weighted to adjust the amount of feed and to evaluate growth. In the end of the experiment, all the shrimp were weighed and counted to determine the survival. Survival did not differ significantly between F. brasiliensis (94.17±9.04) and F. paulensis (98.50±0.71). Although the mean final weight was significantly higher for F. brasiliensis (7.98±0.94g), there were no significant differences in terms of total biomass production (127.81±17.93 e 126.65±1.74g m-2) and apparent feed conversion ratio (1.39±0.27 e 1.57±0.09) between F. brasiliensis and F. paulensis, respectively. The results indicate that F. brasiliensis show potential to be cultured in alternative systems and motivate the development of the technological package for culture of this species in the Patos Lagoon estuary.

Métodos de extrusão manual e elétrica dos espermatóforos de reprodutores selvagens do camarão-rosa Farfantepenaeus paulensis (Decapoda: Penaeidae) / Manual and electrical spermatophore extrusion methods of the pink shrimp Farfantepenaeus paulensis (Decapoda: Penaeidae) wild broodstock

O presente estudo comparou os métodos manual e elétrico de extrusão do espermatóforo do camarão-rosa Farfantepenaeus paulensis com o objetivo de verificar se os métodos de extrusão exercem influência na quantidade de células espermáticas e na regeneração de novos espermatóforos. Os machos foram extrusados no início (dia zero) e no final do experimento (dia 43) para verificação da eficiência dos métodos no processo de regeneração. A extrusão manual foi realizada por meio de pressão na região do quinto par de pereiópodos e o método elétrico com uso de eletrodo para transmissão de impulso elétrico de 9 volts na mesma região. Os dois métodos foram considerados eficientes, não sendo encontradas diferenças significativas entre estes (P>0,05) para o número de células espermáticas. Entretanto, foi verificada, no final do experimento, a perda de peso corporal, peso de espermatóforo e menor índice espermatossomático (IES) (P<0,05) nos animais submetidos a extrusão do espermatóforo por estímulo elétrico. Os valores médios finais (±DP) do número de espermatozóides foram 1,46 (±0,84) e 3,25 milhões (±2,12) para os tratamentos com método elétrico e manual, respectivamente. Os resultados indicam que ambos os métodos podem ser utilizados para a coleta inicial de espermatóforos e testes de qualidade de espermatozóide. Entretanto, para a reutilização dos machos após a extrusão inicial do espermatóforo, o método manual é mais indicado pela manutenção do número de células espermáticas, peso do espermatóforo, peso corporal e índice espermatossomático após a regeneração.

The present study compared manual and electrical methods to extrude the spermatophore of the pink shrimp Farfantepenaeus paulensis aiming to analyze their influence on the number of spermatic cells and spermatophore regeneration. The males were extruded in the beginning (day zero) and in the end (43rd day) of the experiment to evaluate the efficiency of these methods in the regeneration process. For the extrusion, a gentle pressure was applied manually in the fifth pair of pereiopods or electrically by a 9 volt pulse in the same area. Both methods were efficient in removing the spermatophore and no significant differences were found (P>0.05) in the number of sperm cells. Nevertheless, significant decreases (P<0.05) in the body weight, spermatophore weight and spermatosomatic index (ESI) at the end of the experimental period were observed by using the electrical stimulation. The mean values (±SD) of the number of sperm cells were 1.46 (±0.84) and 3.25 (±2.12) millions for the electrical and manual treatments, respectively. Results indicate that both methods may be applied to collect initial samples of spermatophores as well as for sperm quality testing. However, when previously spermatophore-extruded males are to be used, the manual method is indicated as the number of spermatic cells, spermatophore weight, body weight, and ESI are maintained after regeneration.

Shrimp farming as an alternative to artisanal fishermen communities: the Case of Patos Lagoon, Brazil

The shrimp Farfantepenaeus paulensis is considered a suitable species for culture in the Patos Lagoon estuary, southern Brazil, where its catch has been decreasing in the last few years. Considering the potential of estuarine areas for shrimp farming, the application of low-cost structures to culture shrimp, such as the pen enclosures, is being proposed as an alternative source of income for artisanal fishermen and farmers living near Patos Lagoon. From an economical standpoint, these alternative systems present several advantages over the traditional pond-based culture systems since land, water pumping and artificial aerations are not required, high natural productivity is available and inexpensive materials are employed in the construction of grow-out structures. This paper reviews the farming of F. paulensis in the pen enclosures as an alternative to the artisanal fishing communities of the Patos Lagoon estuary. Particular emphases are placed on the development of culture systems, social and environmental impacts, constraints and perspectives.

O camarão-rosa Farfantepenaeus paulensis é considerado uma espécie com potencial para criação no estuário da Lagoa dos Patos, RS, onde o volume desembarcado deste recurso vem decrescendo nos últimos anos. A criação deste camarão em cercados está sendo proposta como uma fonte de renda alternativa para os pescadores artesanais e agricultores que habitam o entorno deste estuário. Do ponto de vista econômico, estes sistemas alternativos de criação apresentam vantagens sobre os sistemas tradicionais em viveiros de terra: não se necessita ter posse da terra, nem equipamentos de bombeamento d'água ou aeração. Além disso, materiais relativamente baratos são utilizados na construção das estruturas de engorda e há uma grande disponibilidade de alimento natural. Esta revisão apresenta alguns aspectos da criação de F. paulensis em cercados como uma alternativa às comunidades de pescadores. Maior ênfase será dada no desenvolvimento dos sistemas de criação, manejo, impactos sociais e ambientais, dificuldades e possibilidades.

Perspectivas para o desenvolvimento dos cultivos de camarões marinhos no estuário da Lagoa dos Patos, RS / Perspectives for the development of marine shrimp farming in the estuary of Patos Lagoon, RS, Brazil

A produção brasileira de camarões vem apresentando um crescimento significativo nos últimos anos, concentrando-se na região nordeste, principal pólo produtor do país. Muito embora as condições climáticas da região Sul não permitam a produção durante todo o ano, uma quantidade considerável de investimentos vem sendo aplicada. No estuário da Lagoa dos Patos, RS, os cultivos da espécie nativa Farfantepenaeus paulensis em cercados e gaiolas e de Litopenaeus vannamei em viveiros estão sendo desenvolvidos como novas atividades, as quais certamente poderão favorecer o desenvolvimento social e econômico da região. Entretanto, a implantação desordenada da carcinicultura pode gerar sérios problemas ambientais. O presente trabalho apresenta uma visão das características ambientais da região em questão e descreve os sistemas de cultivo atualmente utilizados. Além disso, as principais dificuldades enfrentadas por esta nova atividade, assim como a discussão de uma legislação adequada às condições locais e a definição de áreas apropriadas para o cultivo, são apresentadas. Finalmente, o desenvolvimento sustentável da carcinicultura na região é discutido sob as perspectivas social, econômica e ambiental.

Viabilidade do cultivo do camarão-rosa Farfantepenaeus paulensis (Crustácea, Decapoda) em gaiolas sob diferentes densidades durante o outono no sul do Brasil / Viability of culturing the shrimp Farfantepenaeus paulensis (Crustacea, Decapoda) in cages under different stocking densities during autumn in southern Brazil

Neste trabalho, foi estudada a possibilidade de cultivar o camarão-rosa Farfantepenaeus paulensis em gaiolas com diferentes densidades de estocagem durante o outono (abril a junho) no estuário da Lagoa dos Patos, RS. Juvenis com peso médio de 4,95g (n 1,69 dp foram estocados em gaiolas nas densidades de 20, 40, 80, 100 e 120 ind/my. Os camarões foram alimentados diariamente ad libitum com pedacos de Corvina (Micropogonias furnieri). As sobrevivências e os pesos foram observados nos tempos 20, 40 e 60 dias de experimento. Durante o período de cultivo a temperatura da água decresceu de 24 para 11ºC (média = 17,1ºC) e a salinidade oscilou entre 3 e 26 (média = 17,5). As taxas de sobrevivência nas densidades de 20, 40, 80, 100 e 120 ind/my foram 65 por cento, 68 por cento, 46 por cento, 52 por cento e 43,3 por cento respectivamente, sendo significativamente maiores nas densidades de 20 e 40 ind/my. Após 60 dias, os camarões estocados em 20 e 40 ind/my atingiram peso superior a 8,0g, sendo significativamente maiores (P<0,05) que nos demais tratamentos. A maior biomassa (356g) foi obtida na densidade de 120 ind/my. Foram produzidos de 250 a 350g de camarões com peso médio de 6,5 a 8,0g por my, mesmo em altas densidades de estocagem (40 a 120 ind/my). As taxas de sobrevivência dos camarões mantidos em 20 e 40 ind/my podem ser consideradas satisfatórias, contudo, após a temperatura ficar abaixo de 18ºC, as taxas de crescimento foram reduzidas. Os resultados confirmam o efeito negativo da densidade de estocagem sobre o crescimento dos camarões, sendo recomendada a estocagem de 40 a 120 juvenis/my. Foi, contudo, comprovada a possibilidade de cultivar F. paulensis durante o outono no sul do Brasil.

Recent developments on broodstock maturation and reproduction of Farfantepenaeus paulensis

Esta revisão constitui a primeira iniciativa de reunir em um só documento os recentes desenvolvimentos/estudos sobre maturação e reprodução de Farfantepenaeus paulensis em cativeiro. O objetivo do presente estudo foi revisar informações que poderão colaborar para a produção sustentável de náuplios de F. paulensis em cativeiro. Foi dada ênfase aos estudos que vem sendo realizados na última década no Laboratório de Maricultura, Fundação Universidade Federal do Rio Grande, RS. A discussão aborda aspectos relativos ao ciclo reprodutivo, maturação ovariana, sistemas de maturação e desova, inseminação artificial, performance de reprodutores e manutenção de reprodutores em cativeiro.

The influence of water renewal rates on the reproductive and molting cycles of Penaeus paulensis in captivity

The present study analyzed the reproduction of wild-caught Penaeus paulensis in relation to its molt cycle. The experimental design consisted of two treatments (continuous water flow and batch renewal) with two replicates. The stocking density in experimental tanks (1.50 x 0.96m) was approximately 7 animals/m², resulting in 4 males (23.9 ± 2.3 g) and 6 females (52.0 ± 5.5 g) per tank. Shrimp had their uropods cut for individual marking and female maturation was induced through the unilateral eyestalk-ablation. The intermolt period of females and males (17.4 ± 3.2 and 17.8 ± 4.6 days, respectively), number of days between molting and the first spawn (6.9 ± 2.8 days) and number of spawns in the intermolt period (1.4 ± 0.5), presented no significant differences (P>0.05) between treatments. No influences on molt cycle or reproductive performance parameters could be related to differences in water quality parameters, especially nitrogenous compounds, suggesting a trend towards reduce water exchange in shrimp maturation. However, a relative decrease in the number of eggs per spawn was observed. This possibly was due to the smaller maturation tanks. The results of P. paulensis molt cycle could be useful for accompaniment and better planning of the reproduction in captivity