Production of mycelial biomass, proteases and protease inhibitors by Ganoderma lucidum under different submerged fermentation conditions / Produção de biomassa micelial, proteases e inibidores de proteases por Ganoderma lucidum cultivado sob diferentes condições de fermentação submersa

Ganoderma lucidum is a medicinal mushroom widely recognized as a source of biomolecules with pharmacological properties, however, little is known about the factors that influence the synthesis of bioactive proteins by this fungus when cultivated under submerged fermentation. The objective of this work was to evaluate the production of mycelial biomass and intracellular proteases and protease inhibitors by G. lucidum cultivated under different submerged fermentation conditions. The cultivation was carried out in a medium composed of glucose (10 or 20 g.L-1), soy peptone (2.5 or 5 g.L-1) and yeast extract (5 g.L-1), with incubation under agitation (120 rpm) and non-agitation, totaling 8 experimental conditions. Biomass production was determined from the dry weight, while glucose consumption was estimated by quantification of reducing sugars. The proteins were extracted in NaCl (0.15 M), and the protein extracts were submitted to protein quantification by the Bradford method, total proteolytic activity using azocasein, caseinolytic and fibrinolytic activity in Petri dishes, activity of serine (trypsin and chymotrypsin) and cysteine (papain) protease inhibitors. Cultivation in agitated condition showed higher biomass production with a maximum value of 7 g.L-1, in addition to higher activities of trypsin, chymotrypsin and papain inhibitors, with 154 IU.mg-1, 153 IU.mg-1 e 343 IU.mg-1 of protein, respectively. The non-agitated condition showed a greater potential for obtaining proteins, total proteases, caseinolytic and fibrinolytic enzymes, with maximum values of 433 mg.g-1 of extract, 71 U.mL-1 of extract, 63.62 mm2 and 50.27 mm2, respectively. Thus, a medium composed of soy peptone, yest extract and glucose in a 1:2:4 proportion is recommended, under agitation to produce protease inhibitors, and the non-agitated condition when the target is, mainly caseinolytic and fibrinolytic enzymes.

Ganoderma lucidum é um cogumelo medicinal amplamente reconhecido como fonte de biomoléculas com propriedades farmacológicas, entretanto, pouco se conhece acerca dos fatores que influenciam a síntese de proteínas bioativas por esse fungo, quando cultivado sob fermentação submersa. O objetivo deste trabalho foi avaliar a produção de biomassa micelial e de proteases e inibidores de proteases intracelulares por G. lucidum cultivado em diferentes condições de fermentação submersa. O cultivo foi realizado em meio contendo glicose (10 ou 20 g.L-1), peptona de soja (2,5 ou 5 g.L-1) e extrato de levedura (5 g.L-1), com incubação sob agitação (120 rpm) ou não-agitado, totalizando 8 condições experimentais. A produção de biomassa foi determinada a partir do peso seco e o consumo de glicose a partir da quantificação de açúcares redutores. As proteínas foram extraídas em NaCl (0,15 M) e os extratos proteicos foram submetidos à quantificação de proteínas pelo método de Bradford, atividade proteolítica total usando azocaseína, atividade caseinolítica e fibrinolítica em placa de Petri, atividade de inibidores de serino-proteases (tripsina e quimotripsina) e cisteíno-protease (papaína). O cultivo em condição agitada apresentou maior produção de biomassa com valor máximo de 7g.L-1, além de maiores atividades de inibidores de tripsina, quimotripsina e papaína, com 154 UI.mg-1, 153 UI.mg-1 e 343 UI.mg-1 de proteína, respectivamente. A condição não-agitada demonstrou maior potencial para a obtenção de proteínas, proteases totais, enzimas caseinolíticas e fibrinolíticas, com valores máximos de 433 mg.g-1 de extrato, 71 U.mL-1 de extrato, 63,62 mm2 e 50,27 mm2, respectivamente. Assim, recomenda-se o meio composto de peptona de soja, extrato de levedura e glicose na proporção 1:2:4, em condição agitada para a produção de inibidores de proteases, e a condição não-agitada para a síntese de proteases, principalmente enzimas caseinolíticas e fibrinolíticas.

Eurytrema coelomaticum infection: correlation between parasite burden and impairment of pancreatic exocrine enzyme secretion / Eurytrema coelomaticum: correlação entre carga parasitária e insuficiência pancreática exócrina

Eurytrema coelomaticum is a trematode reported in the pancreatic ducts of ruminants. It is conjectured that may cause disorders in the pancreas, as well as digestive and metabolic processes dependent on them. This study, determined if there is an impairment of exocrine pancreatic function, and correlated it with parasite burden. Pancreas, blood, and fecal samples were collected from 119 bovines at a abattoir. Stool samples were subjected to the gelatin and x-ray film digestion tests (to detect the presence of trypsin in feces). Using blood samples, the following biochemical tests were performed: amylase, lipase, glucose, fructosamine, cholesterol, triglycerides, total protein, albumin, and globulins. Analyses were correlated with pancreatic parasite burden. Cattle with a high parasitic load presented higher incidence of negative tests in both gelatin digestion and x-ray film digestion tests (P < 0.001) when compared to non-parasitized animals and those with a low parasitic load. Changes in those tests only occurred if the parasitemia was moderate or severe. The activity of the amylase and lipase enzymes was significantly higher in animals with low parasitemia (P < 0.05), compared to non-parasitized animals and with a high parasitic burden. In this study, in cases of high parasitemia, negative results were observed in both gelatin and x-ray film in the feces digestion tests. However, the low infection of E. coelomaticum, higher levels of serum amylase and lipase that also indicated loss of pancreatic exocrine functions were reported.

Eurytrema coelomaticum, um trematódeo de ductos pancreáticos de ruminantes. Conjectura-se que possa ocasionar transtornos nas funções pancreáticas, mais especificamente nos processos digestivos e metabólicos dependentes destas. Neste estudo, o objetivo foi determinar se há comprometimento da função pancreática exócrina, correlacionado-a a carga parasitária. Foram utilizados pâncreas e respectivas amostras de sangue e fezes de 119 bovinos. As amostras de fezes foram submetidas aos testes de digestão da gelatina em tubo e digestão de filme radiográfico, ambos para detecção de tripsina nas fezes. Foram realizados os seguintes exames bioquímicos em amostras de sangue: amilase, lipase, glicemia, frutosamina, colesterol, triglicerídeos, proteínas totais, albumina e globulinas. Após isto, as análises bioquímicas foram correlacionadas com a quantidade numérica de parasitas encontrados no pâncreas (post-mortem). Houve maior quantidade de testes negativos (digestão do filme radiográfico e prova de digestão da gelatina) nos animais com alta carga parasitária (P < 0.001), quando comparados aos animais não parasitados e com baixa carga parasitária. Portanto, os exames supracitados se alteram somente se a quantidade de parasitas for moderada ou severa. As atividades das enzimas amilase e lipase foram significativamente maiores nos animais que apresentavam baixa parasitemia (P < 0.05), em comparação com os animais com alta carga parasitária e não parasitados. Conclui-se que em quadros de alta parasitemia há alteração significativa nos testes de digestão nas fezes, e que em quadros de baixa parasitemia há alterações significativas nos valores de amilase e lipase séricas, ambos comprovando alterações pancreáticas importantes, de acordo com o quadro de parasitemia.

Quantity - but not diversity - of secreted peptides and proteins increases with age in the tree frog Pithecopus nordestinus

Amphibians inhabit the terrestrial environment, a conquest achieved after several evolutionary steps, which were still insufficient to make them completely independent of the aquatic environment. These processes gave rise to many morphological and physiological changes, making their skin (and cutaneous secretion) rich in bioactive molecules. Among the tree frogs, the secretion is composed mainly of peptides; but alkaloids, proteins and steroids can also be found depending on the species. The most known class of biologically active molecules is the antimicrobial peptides (AMPs) that act against bacteria, fungi and protozoans. Although these molecules are well-studied among the hylids, AMPs ontogeny remains unknown. Therefore, we performed peptidomic and proteomic analyses of Pithecopus nordestinus (formerly Phyllomedusa nordestina) in order to evaluate the peptide content in post-metamorphosed juveniles and adult individuals. Methods: Cutaneous secretion of both life stages of individuals was obtained and analyzed by LC-MS/MS after reduction and alkylation of disulfide bonds or reduction, alkylation and hydrolysis by trypsin. Results: Differences in the TIC profile of juveniles and adults in both treatments were observed. Moreover, the proteomic data revealed known proteins and peptides, with slight differences in the composition, according to the life stage and the treatment. AMPs were identified, and bradykinin-potentiating peptides were observed in trypsin-treated samples, which suggests a protein source of such peptide (cryptide). Conclusion: In general, skin secretion contents were similar between juveniles and adults, varying in quantity, indicating that the different stages of life are reflected in the number of molecules and not on their diversity.(AU)

Quantity - but not diversity - of secreted peptides and proteins increases with age in the tree frog Pithecopus nordestinus

Amphibians inhabit the terrestrial environment, a conquest achieved after several evolutionary steps, which were still insufficient to make them completely independent of the aquatic environment. These processes gave rise to many morphological and physiological changes, making their skin (and cutaneous secretion) rich in bioactive molecules. Among the tree frogs, the secretion is composed mainly of peptides; but alkaloids, proteins and steroids can also be found depending on the species. The most known class of biologically active molecules is the antimicrobial peptides (AMPs) that act against bacteria, fungi and protozoans. Although these molecules are well-studied among the hylids, AMPs ontogeny remains unknown. Therefore, we performed peptidomic and proteomic analyses of Pithecopus nordestinus (formerly Phyllomedusa nordestina) in order to evaluate the peptide content in post-metamorphosed juveniles and adult individuals. Methods: Cutaneous secretion of both life stages of individuals was obtained and analyzed by LC-MS/MS after reduction and alkylation of disulfide bonds or reduction, alkylation and hydrolysis by trypsin. Results: Differences in the TIC profile of juveniles and adults in both treatments were observed. Moreover, the proteomic data revealed known proteins and peptides, with slight differences in the composition, according to the life stage and the treatment. AMPs were identified, and bradykinin-potentiating peptides were observed in trypsin-treated samples, which suggests a protein source of such peptide (cryptide). Conclusion: In general, skin secretion contents were similar between juveniles and adults, varying in quantity, indicating that the different stages of life are reflected in the number of molecules and not on their diversity.(AU)

Hematologic response and serum and peritoneal fluid proteinogram of cattle affected by intestinal diseases and traumatic reticuloperitonitis / Resposta hematológica e proteinograma sérico e do líquido peritoneal de bovinos acometidos com enfermidades intestinais e reticuloperitonite traumática

This study aimed to evaluate the hematologic response and the serum and peritoneal fluid (PF) proteinogram of cattle affected by digestive diseases. Twenty-seven animals were distributed in two groups: GI (intestinal diseases) and GII (traumatic reticuloperitonitis, TRP). The animals were previously submitted to a physical exam. Subsequently, blood samples were collected to perform the complete blood count, determine the plasma protein and fibrinogen, and obtain the serum for proteinogram in polyacrylamide gel (SDS-PAGE). Simultaneously, PF was collected to perform physical and chemical evaluation and the electrophoretic profile (SDS-PAGE). ANOVA at the 5% probability level was used to compare the groups. The animals showed signs of apathy, dehydration, and gastrointestinal hypomotility in both groups. However, GI animals showed more significant clinical changes. The blood count of both groups (P > 0.05) showed leukocytosis due to neutrophilia and a regenerative left shift with hyperfibrinogenemia. The proteinogram of both body fluids allowed the identification of proteins albumin (ALB), transferrin (TRF), ceruloplasmin, haptoglobin, α1-acid glycoprotein (α1-AGP), MW 23000 Da, α1-antitrypsin, IgA, and IgG. The [PT] PF/[PT] blood serum ratio of each of the identified proteins increased, showing statistical differences between groups (P < 0.05) regarding PT, ALB, TRF, α1-AGP, and IgG values, with GI animals showing the highest ratio. Intestinal diseases and TRP triggered a systemic and local response characterized by clinical, hematological, and serum and PF proteinogram alterations. The proteins α1-GPA, haptoglobin, and TRF measured in PF were good inflammation biomarkers and useful as an auxiliary tool for the diagnosis and prognosis of digestive diseases in cattle.(AU)

O objetivo deste estudo foi avaliar a resposta hematológica e o proteinograma sérico e do líquido peritoneal (LP) de bovinos acometidos com doenças digestivas. Foram avaliados 27 bovinos distribuídos em dois grupos, GI (enfermidades intestinais) e GII (reticuloperitonite traumática-RPT). Os animais foram previamente submetidos ao exame físico. Posteriormente foram colhidas amostras de sangue para realização do hemograma, determinação plasmática da proteína e do fibrinogênio e obtenção do soro para realização do proteinograma em gel de poliacrilamida (SDS-PAGE). Simultaneamente foi colhido o LP para avaliação física e química, assim como a realização do perfil eletroforético (SDS-PAGE). Empregou-se a análise de variância ao nível de 5% de probabilidade visando à comparação entre os grupos. Em ambos os grupos os animais demonstraram sinais de apatia, desidratação e hipomotilidade gastrointestinal, no entanto, os animais do GI apresentaram alterações clínicas mais expressivas. No hemograma observou-se em ambos os grupos (P > 0,05) leucocitose por neutrofilia e desvio à esquerda regenerativo com hiperfibrinogenia. O proteinograma de ambos os fluidos corpóreos permitiu a identificação das proteínas albumina (ALB), transferrina (TRF), ceruloplasmina, haptoglobina, α-1 glicoproteína ácida (α1-GPA), PM 23.000 Da, α-1 anti-tripsina, IgA e IgG. Os valores da relação [PT] LP / [PT] soro sanguíneo de cada uma das proteínas identificadas demonstrou elevação dos mesmos, bem como diferença estatística entre grupos (P < 0,05) nos valores da PT, ALB, TRF, α1-GPA e IgG, nos quais a relação foi mais elevada nos animais do GI. As enfermidades intestinais e a RPT desencadearam resposta sistêmica e local caracterizada pelas alterações clínicas, hematológicas, e do proteinograma sérico e do LP. A α1-GPA, a haptoglobina e a TRF, mensuradas no LP se mostraram bons biomarcadores de inflamação, sendo úteis como recurso auxiliar de diagnóstico e prognóstico das doenças digestivas dos bovinos.(AU)

Partial characterization of digestive proteases in juveniles of Microphis brachyurus (short-tailed pipefish) (Syngnathiformes: Syngnathidae)

RESUMEN El pez pipa (Microphis brachyurus) es un organismo dulceacuícola con alto potencial económico para la acuarofilia; sin embargo, es necesario implementar su cultivo a través de estudios de fisiología digestiva. Se evaluó el efecto del pH, temperatura e inhibidores sobre las actividades enzimáticas de proteasas ácidas y alcalinas. El pH óptimo de proteasas estomacales es de 2, mientras que el de proteases intestinales es de 10. La temperatura óptima de proteasas ácidas es de 35 ºC y las alcalinas de 45 ºC. La estabilidad térmica para proteasas ácidas y alcalinas es a los 35 ºC (más de 100% de actividad residual). La estabilidad a los diferentes pH de las proteasas ácidas es en 2 (50 % de la actividad residual), mientras que para las proteasas alcalinas es en 10 (90 % de la actividad residual). Las proteasas ácidas fueron inhibidas en 80% con pepstatina A y las proteasas alcalinas fueron altamente inhibidas con TLCK para tripsina (75%) y TPCK quimitripsina (80%). Finalmente, las metaloproteasas fueron inactivadas con EDTA en 70%. En conclusión, M. brachyurus tiene una buena capacidad digestiva al degradar una amplia variedad de proteinas debido a su alta actividad proteolítica.

ABSTRACT Short-tailed pipe fish (Microphis brachyurus) is a freshwater organism with high economic potential for the aquarium hobby, so it is necessary to implement methods to promote its culture through studies of digestive physiology. General activities of acid and alkaline proteases were evaluated, as well as the effect of pH, temperature and inhibitors. The optimal pH of stomach proteases was 2, while the optimal pH of intestinal proteases was 10. Optimal temperature for the acidic proteases was 35 ºC, while for alkaline proteases it was 45 ºC. Thermal stability showed high resistance at 35 ºC for both acid and alkaline proteases (above 100% residual activity). Acid proteases are resistant at pH 2 (50% of residual activity), meanwhile alkaline proteases were highly resistant at pH 10 (90% of residual activity). Acid proteases were inhibited by 80% with pepstatin A and alkaline proteases were inhibited with TLCK and TPCK for trypsin (75%) and chymotrypsin (80%), respectively. Finally, metallo-proteases were 75% partially inhibited some serine proteases by 75% with EDTA. In conclusion, M. brachyurus has a good digestive capacity, since they can degrade a wide variety of proteins due to their greater proteolytic activity.

Citral as food additive for common snook - zootechnical parameters and digestive enzymes / Citral como aditivo na ração para robalo flecha - parâmetros zootécnicos e enzimas digestivas

Essential oils of plants whose main compound is citral showed beneficial effects when added to fish feed. The objective of the present study was to evaluate the dietary effect of the addition of citral on zootechnical parameters and digestive enzyme activities of Centropomus undecimalis. Juveniles were fed for 45 days with diets containing different amounts of citral (0.0 - control, 0.5, 1.0, and 2.0 mL per kg of diet). The water quality parameters were kept stable during the experiment and no mortality was observed. At the end of the experimental period, the treatment 0.5 mL citral per kg of diet had the lowest weight gain and specific growth rate, and the highest feed conversion, while the same parameters did not differ between the other treatments. Pepsin activity was higher in the stomach of fish fed with 0.5 mL citral per kg of diet and amylase activity was higher in the intestine of fish fed with 0.5 and 2.0 mL citral per kg of diet compared to the control group. Intestinal lipase activity was higher in all groups that were fed with citral compared to the control group. Chymotrypsin and trypsin activities showed no difference between groups. Consequently, dietary addition of citral at any of the levels tested is not recommended for common snook.(AU)

Óleos essenciais de plantas, cujo composto majoritário é o citral, mostraram efeitos benéficos quando adicionados à ração para peixes. O objetivo do presente estudo foi avaliar o efeito da adição de citral sobre os parâmetros zootécnicos e atividades das enzimas digestivas de Centropomus undecimalis. Os juvenis foram alimentados por 45 dias com dietas contendo diferentes quantidades de citral (0,0 - controle, 0,5, 1,0 e 2,0 mL por kg de ração). Os parâmetros de qualidade da água foram mantidos estáveis ​​durante o experimento e nenhuma mortalidade foi observada. Ao final do período experimental, o tratamento 0,5 mL citral por kg de ração teve o menor ganho de peso e taxa de crescimento específico e a maior conversão alimentar, enquanto os mesmos parâmetros não diferiram entre os demais tratamentos. A atividade da pepsina foi maior no estômago de peixes alimentados com 0,5 mL de citral por kg de ração e a atividade de amilase foi maior no intestino de peixes alimentados com 0,5 e 2,0 mL citral por kg de ração comparado ao grupo controle. A atividade da lipase intestinal foi maior em todos os grupos que foram alimentados com citral, em comparação ao grupo controle. As atividades de quimotripsina e tripsina não mostraram diferença entre os grupos. Consequentemente, a adição de citral na ração em qualquer um dos níveis testados não é recomendada para robalo flecha.(AU)

Effect of replacing fish meal by full fat soybean meal on growth performance, feed utilization and gastrointestinal enzymes in diets for African catfish Clarias gariepinus / Efeito da substituição de farinha de peixe por farelo de soja integral no desempenho de crescimento, utilização de ração e enzimas gastrointestinais em dietas para bagre africano Clarias gariepinus

Study aimed to evaluate the effect of replacing fish meal with different levels of full fat soybean meal (FFSBM) on growth and digestive enzyme activities in the stomach, Liver and intestine for Clarias gariepinus. Four diets (D1, D2, D3 and D4) were formulated with 0, 15, 20 and 20 g 100-1 protein + DL-methionine by alternating FFSBM with fish meal. The growth of C. gariepinus was found to be significantly decreased when FFSBM replacement increased. Final body weight was 89.69, 79.70, 70.82 and 68.29 g for fish fed on D1, D2, D3 and D4, respectively, with significant differences between treatments. Specific growth rate (SGR) ranged between 3.11 to 2.78%. Proteolytic activity was higher only with alkaline pHs, whereas only very low activity was shown with acidic. Results of liver showed approximately similar results at acid and alkaline. In contrast, higher proteolytic activity in the stomach was observed at acid pHs 3.0 and 4.0 g tyrosine-1 minute-1 mg-1 protein) whereas lower values were observed at neutral pH 7.0 g tyrosine-1 minute-1 mg-1 protein for catfish fed on the experimental diets. Moreover, trypsin activity was higher for the stomach, followed by the intestine and liver. However, higher amount of amylase observed in the liver than intestine and stomach.(AU)

Objetivou-se avaliar o efeito da substituição da farinha de peixe por diferentes níveis de farelo de soja integral (FSI) sobre o crescimento e atividades de enzimas digestivas no estômago, fígado e intestino de Clarias gariepinus. Quatro dietas (D1, D2, D3 e D4) foram formuladas com 0, 15, 20 e 20 g 100-1 proteína + DL-metionina, alternando FSI com farinha de peixe. O crescimento de C. gariepinus foi significativamente reduzido quando aumentou a substituição de farinha de peixe por FSI . O peso corporal final foi de 89,69, 79,70, 70,82 e 68,29 g para peixes alimentados com D1, D2, D3 e D4, respectivamente, com diferenças significativas entre os tratamentos. Taxa de crescimento específico (TCE) variou entre 3,11 a 2,78%. A atividade proteolítica foi maior apenas com pHs alcalinos, enquanto que atividades muito baixas foram somente mostradas com ácido. Os resultados no fígado mostraram valores aproximadamente semelhantes tanto em meio ácido quanto alcalino. Em contraste, observou-se maior atividade proteolítica no estômago em pH ácido (3,0 e 4,0 g de tirosina-1 minuto-1 mg-1 de proteína), ao passo que valores mais baixos foram observados em pH neutro (7,0 g de tirosina-1 minuto-1 mg-1 de proteína) para os bagres que se alimentaram com as dietas experimentais. Além disso, a atividade da tripsina foi maior para o estômago, seguida pelo intestino e pelo fígado. No entanto, observou-se maior quantidade de amilase no fígado,intestino e estômago.(AU)

Partial characterization of digestive proteases in juveniles of Microphis brachyurus (short-tailed pipefish) (Syngnathiformes: Syngnathidae)

Short-tailed pipe fish (Microphis brachyurus) is a freshwater organism with high economic potential for the aquarium hobby, so it is necessary to implement methods to promote its culture through studies of digestive physiology. General activities of acid and alkaline proteases were evaluated, as well as the effect of pH, temperature and inhibitors. The optimal pH of stomach proteases was 2, while the optimal pH of intestinal proteases was 10. Optimal temperature for the acidic proteases was 35 ºC, while for alkaline proteases it was 45 ºC. Thermal stability showed high resistance at 35 ºC for both acid and alkaline proteases (above 100% residual activity). Acid proteases are resistant at pH 2 (50% of residual activity), meanwhile alkaline proteases were highly resistant at pH 10 (90% of residual activity). Acid proteases were inhibited by 80% with pepstatin A and alkaline proteases were inhibited with TLCK and TPCK for trypsin (75%) and chymotrypsin (80%), respectively. Finally, metallo-proteases were 75% partially inhibited some serine proteases by 75% with EDTA. In conclusion, M. brachyurus has a good digestive capacity, since they can degrade a wide variety of proteins due to their greater proteolytic activity.(AU)

El pez pipa (Microphis brachyurus) es un organismo dulceacuícola con alto potencial económico para la acuarofilia; sin embargo, es necesario implementar su cultivo a través de estudios de fisiología digestiva. Se evaluó el efecto del pH, temperatura e inhibidores sobre las actividades enzimáticas de proteasas ácidas y alcalinas. El pH óptimo de proteasas estomacales es de 2, mientras que el de proteases intestinales es de 10. La temperatura óptima de proteasas ácidas es de 35 ºC y las alcalinas de 45 ºC. La estabilidad térmica para proteasas ácidas y alcalinas es a los 35 ºC (más de 100% de actividad residual). La estabilidad a los diferentes pH de las proteasas ácidas es en 2 (50 % de la actividad residual), mientras que para las proteasas alcalinas es en 10 (90 % de la actividad residual). Las proteasas ácidas fueron inhibidas en 80% con pepstatina A y las proteasas alcalinas fueron altamente inhibidas con TLCK para tripsina (75%) y TPCK quimitripsina (80%). Finalmente, las metaloproteasas fueron inactivadas con EDTA en 70%. En conclusión, M. brachyurus tiene una buena capacidad digestiva al degradar una amplia variedad de proteinas debido a su alta actividad proteolítica.(AU)

Changes in digestive enzymes activities during the initial ontogeny of wolf cichlid, Parachromis dovii (Perciformes: Cichlidae)

Wolf cichlid, Parachromis dovii, is a species with a high potential for aquaculture in Central America; however, the knowledge of the digestive physiology in larvae period is limited. For these reason, this study evaluated the changes on digestive enzymes (alkaline and acid proteases, trypsin, chymotrypsin, aminopeptidase, carboxypeptidase, lipases, amylases, and phosphatases) during early ontogeny by biochemical analysis. All digestive enzymes were detected at first feeding (6 days after hatching, DAH, 9.49 mm, 168 degree-days DD). Afterwards all enzymes reached two main peaks in activity at 14 or 22 DAH (15.10 mm, 364 DD and 20.83 mm, 550 DD, respectively). Later, there was a gradual decrease in activity for trypsin and acid and alkaline phosphatases until reach the lowest values at 41 DAH. In the case of acid proteases, chymotrypsin, aminopeptidase, carboxypeptidase, lipase and amylase, all activities reached their maximum values at the end of the larval period, except for alkaline proteases, which showed the maximum value at 14 DAH (15.10 mm, 364 DD). Parachromis dovii larvae have an early capability to hydrolyze exogenous food, agreeing with other carnivorous neotropical cichlid species, for this reason we proposed that the weaning process could begin at 14 DAH.(AU)

El guapote lagunero (Parachromis dovii) es una especie con un alto potencial para la acuicultura en la región de América Central; sin embargo, existe un conocimiento limitado sobre la capacidad digestiva en el periodo larval. Por este motivo, este estudio evaluó los cambios de las enzimas digestivas (proteasas alcalinas y ácidas, tripsina, quimotripsina, aminopeptidasa, carboxipeptidasa, lipasas, amilasas y fosfatasas) durante la ontogenia temprana mediante análisis bioquímico. Todas las enzimas digestivas analizadas se detectaron en la primera alimentación (6 días después de la eclosión, DAH, 9.49 mm, 168 día-grados DD). Después, todas las enzimas alcanzaron dos picos máximos a los 14 o 22 DAH (15.10 mm, 364 DD and 20.83 mm, 550 DD, respectivamente). Después las actividades tripsina, fosfatasas ácidas y alcalina disminuyeron a sus valores más bajos a los 41 DAH. En el caso de las proteasas ácidas y alcalinas, quimotripsina, aminopeptidasa, carboxipeptidasa, lipasa y amilasa, los niveles de actividad aumentaron y alcanzaron su máximo valor al final del período larvario, excepto las proteasas alcalinas, que mostraron su máximo valor a los 14 DAH (15.10 mm, 364 DD). Las larvas de P. dovii tienen una capacidad temprana para hidrolizar alimentos exógenos, lo que concuerda con otras especies de cíclidos neotropicales carnívoros, por lo que proponemos que el proceso de destete inicie a los 14 DAH.(AU)

Changes in digestive enzymes activities during the initial ontogeny of wolf cichlid, Parachromis dovii (Perciformes: Cichlidae)

Wolf cichlid, Parachromis dovii, is a species with a high potential for aquaculture in Central America; however, the knowledge of the digestive physiology in larvae period is limited. For these reason, this study evaluated the changes on digestive enzymes (alkaline and acid proteases, trypsin, chymotrypsin, aminopeptidase, carboxypeptidase, lipases, amylases, and phosphatases) during early ontogeny by biochemical analysis. All digestive enzymes were detected at first feeding (6 days after hatching, DAH, 9.49 mm, 168 degree-days DD). Afterwards all enzymes reached two main peaks in activity at 14 or 22 DAH (15.10 mm, 364 DD and 20.83 mm, 550 DD, respectively). Later, there was a gradual decrease in activity for trypsin and acid and alkaline phosphatases until reach the lowest values at 41 DAH. In the case of acid proteases, chymotrypsin, aminopeptidase, carboxypeptidase, lipase and amylase, all activities reached their maximum values at the end of the larval period, except for alkaline proteases, which showed the maximum value at 14 DAH (15.10 mm, 364 DD). Parachromis dovii larvae have an early capability to hydrolyze exogenous food, agreeing with other carnivorous neotropical cichlid species, for this reason we proposed that the weaning process could begin at 14 DAH.(AU)

El guapote lagunero (Parachromis dovii) es una especie con un alto potencial para la acuicultura en la región de América Central; sin embargo, existe un conocimiento limitado sobre la capacidad digestiva en el periodo larval. Por este motivo, este estudio evaluó los cambios de las enzimas digestivas (proteasas alcalinas y ácidas, tripsina, quimotripsina, aminopeptidasa, carboxipeptidasa, lipasas, amilasas y fosfatasas) durante la ontogenia temprana mediante análisis bioquímico. Todas las enzimas digestivas analizadas se detectaron en la primera alimentación (6 días después de la eclosión, DAH, 9.49 mm, 168 día-grados DD). Después, todas las enzimas alcanzaron dos picos máximos a los 14 o 22 DAH (15.10 mm, 364 DD and 20.83 mm, 550 DD, respectivamente). Después las actividades tripsina, fosfatasas ácidas y alcalina disminuyeron a sus valores más bajos a los 41 DAH. En el caso de las proteasas ácidas y alcalinas, quimotripsina, aminopeptidasa, carboxipeptidasa, lipasa y amilasa, los niveles de actividad aumentaron y alcanzaron su máximo valor al final del período larvario, excepto las proteasas alcalinas, que mostraron su máximo valor a los 14 DAH (15.10 mm, 364 DD). Las larvas de P. dovii tienen una capacidad temprana para hidrolizar alimentos exógenos, lo que concuerda con otras especies de cíclidos neotropicales carnívoros, por lo que proponemos que el proceso de destete inicie a los 14 DAH.(AU)

Effect of replacing fish meal by full fat soybean meal on growth performance, feed utilization and gastrointestinal enzymes in diets for African catfish Clarias gariepinus

Abstract Study aimed to evaluate the effect of replacing fish meal with different levels of full fat soybean meal (FFSBM) on growth and digestive enzyme activities in the stomach, Liver and intestine for Clarias gariepinus. Four diets (D1, D2, D3 and D4) were formulated with 0, 15, 20 and 20 g 100-1 protein + DL-methionine by alternating FFSBM with fish meal. The growth of C. gariepinus was found to be significantly decreased when FFSBM replacement increased. Final body weight was 89.69, 79.70, 70.82 and 68.29 g for fish fed on D1, D2, D3 and D4, respectively, with significant differences between treatments. Specific growth rate (SGR) ranged between 3.11 to 2.78%. Proteolytic activity was higher only with alkaline pHs, whereas only very low activity was shown with acidic. Results of liver showed approximately similar results at acid and alkaline. In contrast, higher proteolytic activity in the stomach was observed at acid pHs 3.0 and 4.0 g tyrosine-1 minute-1 mg-1 protein) whereas lower values were observed at neutral pH 7.0 g tyrosine-1 minute-1 mg-1 protein for catfish fed on the experimental diets. Moreover, trypsin activity was higher for the stomach, followed by the intestine and liver. However, higher amount of amylase observed in the liver than intestine and stomach.

Resumo Objetivou-se avaliar o efeito da substituição da farinha de peixe por diferentes níveis de farelo de soja integral (FSI) sobre o crescimento e atividades de enzimas digestivas no estômago, fígado e intestino de Clarias gariepinus. Quatro dietas (D1, D2, D3 e D4) foram formuladas com 0, 15, 20 e 20 g 100-1 proteína + DL-metionina, alternando FSI com farinha de peixe. O crescimento de C. gariepinus foi significativamente reduzido quando aumentou a substituição de farinha de peixe por FSI . O peso corporal final foi de 89,69, 79,70, 70,82 e 68,29 g para peixes alimentados com D1, D2, D3 e D4, respectivamente, com diferenças significativas entre os tratamentos. Taxa de crescimento específico (TCE) variou entre 3,11 a 2,78%. A atividade proteolítica foi maior apenas com pHs alcalinos, enquanto que atividades muito baixas foram somente mostradas com ácido. Os resultados no fígado mostraram valores aproximadamente semelhantes tanto em meio ácido quanto alcalino. Em contraste, observou-se maior atividade proteolítica no estômago em pH ácido (3,0 e 4,0 g de tirosina-1 minuto-1 mg-1 de proteína), ao passo que valores mais baixos foram observados em pH neutro (7,0 g de tirosina-1 minuto-1 mg-1 de proteína) para os bagres que se alimentaram com as dietas experimentais. Além disso, a atividade da tripsina foi maior para o estômago, seguida pelo intestino e pelo fígado. No entanto, observou-se maior quantidade de amilase no fígado,intestino e estômago.

Non-starch polysaccharide in diets for hybrid surubim: effects on metabolism plasticity / Polissacarídeos não amiláceos em dietas para híbrido de surubim: efeitos sobre a plasticidade do metabolismo

This study aimed to analyze the effect of mango meal, a non-starch polysaccharide, as a carbohydrate source on the metabolism plasticity and performance indicators for hybrid surubim (Pseudoplatystoma fasciatum x P. corruscans), a tropical carnivorous fish. Juveniles (±14.5g initial weight) were fed 3% of total biomass for 60 days in four experimental diets replacing 0, 33, 66 and 100% of the cornmeal. Growth by final weight was evaluated, but statistical differences (p<0.05) were not observed among the treatments. The activity of total alkaline protease was statistically lower in the fish group fed 100% cornmeal substitution. However, trypsin levels also decreased when 100% of the cornmeal was replaced by mango meal, but without statistical significance. Zymograms showed bands of protease and amylase activities for all treatments. Cholesterol (0%- 141.83±46.10; 100%- 71.36±14.40 mg.dL-1) and total protein (0%- 3.98±0.94; 100%- 1.70±1.03 mg.dL-1) in the plasma decreased as mango meal levels increased in the feed. On the other hand, glucose (0%- 105.08±31.24; 100%- 128.11±24.51 mg.dL-1) and free amino acids (0%- 24.51±2.62; 100%- 38.13±8.94 nmoles.ml-1) in the plasma increased. Although growth was not affected, the carbohydrate source affected fish metabolism, which showed metabolic adaptation by the species. The result suggests that mango meal can replace up to 33% of the cornmeal in diets for Pseudoplatystoma sp.

O objetivo deste estudo foi avaliar o efeito da farinha de manga, um polissacarídeos não amiláceo, como fonte de carboidrato na plasticidade do metabolismo e indicadores de desempenho no surubim híbrido (Pseudoplatystoma fasciatum x P. corruscans) um peixe carnívoro tropical. Juvenis (±14.5g peso inicial) foram alimentados com 3% da biomassa total durante 60 dias, com 4 dietas experimentais substituindo 0, 33, 66 e 100% do farelo de milho. O crescimento final foi avaliado, mas diferenças significativas (p<0.05) não foram observadas entre os tratamentos. A atividade da protease total alcalina foi significativamente menor no tratamento onde houve 100% de substituição do farelo de milho. Entretanto, níveis de tripsina também diminuíram quando 100% do farelo de milho foi substituído por farinha de manga, mas sem diferença significativa. O zimograma mostrou bandas de atividade de protease e amilase em todos os tratamentos. Colesterol (0%- 141.83±46.10; 100%- 71.36±14.40 mg.dL-1) e proteína total (0%- 3.98±0.94; 100%- 1.70±1.03 mg.dL-1) plasmáticos, diminuíram à medida que os níveis de farinha de manga aumentaram nas dietas. Por outro lado, glicose (0%- 105.08±31.24; 100%- 128.11±24.51 mg.dL-1) e aminoácidos livres (0%- 24.51±2.62; 100%- 38.13±8.94 nmoles.ml-1) no plasma, aumentaram. Embora o crescimento não foi aumentado, a fonte de carboidrato afetou o metabolismo do animal, o que mostra adaptação metabólica do híbrido. O resultado sugere que farinha de manga pode substituir até 33% de farelo de milho em dietas para Pseudoplatystoma sp.

Growth and digestive enzymes of silver catfish fed diets containing fruit residue / Crescimento e enzimas digestivas de jundiás alimentados com dietas contendo resíduos de frutas

In fish farming, the use of alternative ingredients has been studied, so that alternative sources can be used to minimize feed costs. This study evaluated the incorporation of grape, orange, guava, and fig residues in diets for silver catfish and its effects on growth, digestive enzymes and body composition. A total of 180 fish (initial mean weight = 22.93±0.75 g) were reared in a recirculation aquaculture system. There was no difference (P>0.05) in the parameters of growth, dry matter, mineral matter, plasma protein, cholesterol, triglycerides, lipase, and trypsin of fish. Glucose levels were higher in fish fed diets containing fig, orange, and grape residue (P<0.05). Lipase activity was higher in fish fed orange residue, compared to guava (P<0.05). Diets containing guava and fig provided more body protein in silver catfish. The fish fed with diet containing orange residue had a higher content of body lipids. It can be concluded that the tested fruit residues can be used in silver catfish feeding.(AU)

Na piscicultura, o uso de ingredientes alternativos vem sendo estudado, de modo que seja possível utilizar fontes alternativas para minimizar custos com a alimentação. Neste estudo, foi avaliada a incorporação de resíduos de uva, laranja, goiaba e figo em rações para jundiá e seus efeitos no crescimento, em enzimas digestivas e na composição corporal. Foram utilizados 180 peixes (peso médio inicial = 22,93±0,75 g), criados em sistema de recirculação. Não houve diferença (P>0,05) nos parâmetros de crescimento, matéria seca, matéria mineral, proteína plasmática, colesterol, triglicerídeos, lipase e tripsina dos peixes. Os níveis de glicose foram maiores nos peixes alimentados com dietas contendo figo, laranja e uva (P<0,05). A atividade de lipase foi maior nos peixes alimentados com resíduo de laranja, comparado com o de goiaba (P<0,05). As dietas contendo goiaba e figo proporcionaram mais proteína corporal nos jundiás. Os jundiás alimentados com dieta contendo resíduo de laranja apresentaram maior teor de lipídeos corporal. Conclui-se que os resíduos de frutas testados podem ser usados na alimentação do jundiá.(AU)

Growth and digestive enzymes of silver catfish fed diets containing fruit residue / Crescimento e enzimas digestivas de jundiás alimentados com dietas contendo resíduos de frutas

In fish farming, the use of alternative ingredients has been studied, so that alternative sources can be used to minimize feed costs. This study evaluated the incorporation of grape, orange, guava, and fig residues in diets for silver catfish and its effects on growth, digestive enzymes and body composition. A total of 180 fish (initial mean weight = 22.93±0.75 g) were reared in a recirculation aquaculture system. There was no difference (P>0.05) in the parameters of growth, dry matter, mineral matter, plasma protein, cholesterol, triglycerides, lipase, and trypsin of fish. Glucose levels were higher in fish fed diets containing fig, orange, and grape residue (P<0.05). Lipase activity was higher in fish fed orange residue, compared to guava (P<0.05). Diets containing guava and fig provided more body protein in silver catfish. The fish fed with diet containing orange residue had a higher content of body lipids. It can be concluded that the tested fruit residues can be used in silver catfish feeding.(AU)

Na piscicultura, o uso de ingredientes alternativos vem sendo estudado, de modo que seja possível utilizar fontes alternativas para minimizar custos com a alimentação. Neste estudo, foi avaliada a incorporação de resíduos de uva, laranja, goiaba e figo em rações para jundiá e seus efeitos no crescimento, em enzimas digestivas e na composição corporal. Foram utilizados 180 peixes (peso médio inicial = 22,93±0,75 g), criados em sistema de recirculação. Não houve diferença (P>0,05) nos parâmetros de crescimento, matéria seca, matéria mineral, proteína plasmática, colesterol, triglicerídeos, lipase e tripsina dos peixes. Os níveis de glicose foram maiores nos peixes alimentados com dietas contendo figo, laranja e uva (P<0,05). A atividade de lipase foi maior nos peixes alimentados com resíduo de laranja, comparado com o de goiaba (P<0,05). As dietas contendo goiaba e figo proporcionaram mais proteína corporal nos jundiás. Os jundiás alimentados com dieta contendo resíduo de laranja apresentaram maior teor de lipídeos corporal. Conclui-se que os resíduos de frutas testados podem ser usados na alimentação do jundiá.(AU)

Non-starch polysaccharide in diets for hybrid surubim: effects on metabolism plasticity / Polissacarídeos não amiláceos em dietas para híbrido de surubim: efeitos sobre a plasticidade do metabolismo

This study aimed to analyze the effect of mango meal, a non-starch polysaccharide, as a carbohydrate source on the metabolism plasticity and performance indicators for hybrid surubim (Pseudoplatystoma fasciatum x P. corruscans), a tropical carnivorous fish. Juveniles (±14.5g initial weight) were fed 3% of total biomass for 60 days in four experimental diets replacing 0, 33, 66 and 100% of the cornmeal. Growth by final weight was evaluated, but statistical differences (p<0.05) were not observed among the treatments. The activity of total alkaline protease was statistically lower in the fish group fed 100% cornmeal substitution. However, trypsin levels also decreased when 100% of the cornmeal was replaced by mango meal, but without statistical significance. Zymograms showed bands of protease and amylase activities for all treatments. Cholesterol (0%- 141.83±46.10; 100%- 71.36±14.40 mg.dL-1) and total protein (0%- 3.98±0.94; 100%- 1.70±1.03 mg.dL-1) in the plasma decreased as mango meal levels increased in the feed. On the other hand, glucose (0%- 105.08±31.24; 100%- 128.11±24.51 mg.dL-1) and free amino acids (0%- 24.51±2.62; 100%- 38.13±8.94 nmoles.ml-1) in the plasma increased. Although growth was not affected, the carbohydrate source affected fish metabolism, which showed metabolic adaptation by the species. The result suggests that mango meal can replace up to 33% of the cornmeal in diets for Pseudoplatystoma sp.(AU)

O objetivo deste estudo foi avaliar o efeito da farinha de manga, um polissacarídeos não amiláceo, como fonte de carboidrato na plasticidade do metabolismo e indicadores de desempenho no surubim híbrido (Pseudoplatystoma fasciatum x P. corruscans) um peixe carnívoro tropical. Juvenis (±14.5g peso inicial) foram alimentados com 3% da biomassa total durante 60 dias, com 4 dietas experimentais substituindo 0, 33, 66 e 100% do farelo de milho. O crescimento final foi avaliado, mas diferenças significativas (p<0.05) não foram observadas entre os tratamentos. A atividade da protease total alcalina foi significativamente menor no tratamento onde houve 100% de substituição do farelo de milho. Entretanto, níveis de tripsina também diminuíram quando 100% do farelo de milho foi substituído por farinha de manga, mas sem diferença significativa. O zimograma mostrou bandas de atividade de protease e amilase em todos os tratamentos. Colesterol (0%- 141.83±46.10; 100%- 71.36±14.40 mg.dL-1) e proteína total (0%- 3.98±0.94; 100%- 1.70±1.03 mg.dL-1) plasmáticos, diminuíram à medida que os níveis de farinha de manga aumentaram nas dietas. Por outro lado, glicose (0%- 105.08±31.24; 100%- 128.11±24.51 mg.dL-1) e aminoácidos livres (0%- 24.51±2.62; 100%- 38.13±8.94 nmoles.ml-1) no plasma, aumentaram. Embora o crescimento não foi aumentado, a fonte de carboidrato afetou o metabolismo do animal, o que mostra adaptação metabólica do híbrido. O resultado sugere que farinha de manga pode substituir até 33% de farelo de milho em dietas para Pseudoplatystoma sp.(AU)

Resistance of maize cultivars to Sitophilus zeamais (Coleoptera: Curculionidae) / Resistência de cultivares de milho a Sitophilus zeamais (Coleoptera: Curculionidae)

Five Zea mays cultivars (BRS Caatingueiro, BRS Gorutuba, BRS Sertanejo, BRS Asa Branca and BR 106) were evaluated considering their effect on the nutrition of the maize weevil Sitophilus zeamais, by analysis of total protein in adult fed with these cultivars and for the presence of lectins and trypsin inhibitors in grains. In addition, free-choice and no-choice assays were performed to investigate the resistance of grains of the Z. mays cultivars to an attack by S. zeamais. The BR 106 cultivar showed the lowest susceptibility index, followed by BRS Caatingueiro, BRS Asa Branca, BRS Sertanejo and BRS Gorutuba. The number of emerged adults in the Z. mays cultivars ranged from 213.17 to 74.0, and the lowest number of insects was recorded for the BR 106 cultivar. The insects were able to feed on grains of all cultivars, but the BR 106 cultivar showed the least reduction in dried biomass. Lectins were detected in extracts from BR 106, BRS Asa Branca, BRS Sertanejo and BRS Gorutuba, and the highest activity was shown by BR 106. The lowest protein assimilation was detected in the insects from treatments with BRS Asa Branca. The extracts from all cultivars were able to inhibit the activity of bovine trypsin, but this effect was not related to the resistance degree of Z. mays cultivars. The results suggest the resistance of BR 160 to the attack of S. zeamais, as well as indicating that the presence of lectin in the grains is the cause of this resistance.(AU)

Foram avaliadas cinco cultivares de Zea mays (BRS Caatingueiro, BRS Gorutuba, BRS Sertanejo, BRS Asa Branca e BR 106) e seu efeito na nutrição do gorgulho-do-milho Sitophilus zeamais, por meio da análise de proteína total em adultos alimentados com esses cultivares e a presença de lectinas e inibidores da tripsina nos grãos. Além disso, foram realizados ensaios com e sem chance de escolha para investigar a resistência dos cultivares de Z. mays ao ataque de S. zeamais. O cultivar BR 106 apresentou o menor índice de susceptibilidade, seguido por BRS Caatingueiro, BRS Asa Branca, BRS Sertanejo e BRS Gorutuba. O número de adultos emergidos nos cultivares de Z. mays variou de 213,17 a 74,0, e o menor número de insetos foi registrado para o cultivar BR 106. Os insetos foram capazes de se alimentar de todos os cultivares, no entanto, o BR 106 mostrou a menor redução na biomassa seca. As lectinas foram detectadas em extratos de BR 106, BRS Asa Branca, BRS Sertanejo e BRS Gorutuba, e a maior atividade foi demonstrada pela BR 106. A menor assimilação de proteína foi detectada nos insetos que se alimentaram com BRS Asa Branca. Os extratos de todos os cultivares foram capazes de inibir a atividade da tripsina bovina, mas esse efeito não está correlacionado ao grau de resistência dos cultivares de Z. mays. Os resultados sugerem a resistência da BR 160 ao ataque de S. zeamais, além de indicar que a presença de lectina nos grãos é a causa dessa resistência.(AU)

Resistance of maize cultivars to Sitophilus zeamais (Coleoptera: Curculionidae) / Resistência de cultivares de milho a Sitophilus zeamais (Coleoptera: Curculionidae)

Five Zea mays cultivars (BRS Caatingueiro, BRS Gorutuba, BRS Sertanejo, BRS Asa Branca and BR 106) were evaluated considering their effect on the nutrition of the maize weevil Sitophilus zeamais, by analysis of total protein in adult fed with these cultivars and for the presence of lectins and trypsin inhibitors in grains. In addition, free-choice and no-choice assays were performed to investigate the resistance of grains of the Z. mays cultivars to an attack by S. zeamais. The BR 106 cultivar showed the lowest susceptibility index, followed by BRS Caatingueiro, BRS Asa Branca, BRS Sertanejo and BRS Gorutuba. The number of emerged adults in the Z. mays cultivars ranged from 213.17 to 74.0, and the lowest number of insects was recorded for the BR 106 cultivar. The insects were able to feed on grains of all cultivars, but the BR 106 cultivar showed the least reduction in dried biomass. Lectins were detected in extracts from BR 106, BRS Asa Branca, BRS Sertanejo and BRS Gorutuba, and the highest activity was shown by BR 106. The lowest protein assimilation was detected in the insects from treatments with BRS Asa Branca. The extracts from all cultivars were able to inhibit the activity of bovine trypsin, but this effect was not related to the resistance degree of Z. mays cultivars. The results suggest the resistance of BR 160 to the attack of S. zeamais, as well as indicating that the presence of lectin in the grains is the cause of this resistance.(AU)

Foram avaliadas cinco cultivares de Zea mays (BRS Caatingueiro, BRS Gorutuba, BRS Sertanejo, BRS Asa Branca e BR 106) e seu efeito na nutrição do gorgulho-do-milho Sitophilus zeamais, por meio da análise de proteína total em adultos alimentados com esses cultivares e a presença de lectinas e inibidores da tripsina nos grãos. Além disso, foram realizados ensaios com e sem chance de escolha para investigar a resistência dos cultivares de Z. mays ao ataque de S. zeamais. O cultivar BR 106 apresentou o menor índice de susceptibilidade, seguido por BRS Caatingueiro, BRS Asa Branca, BRS Sertanejo e BRS Gorutuba. O número de adultos emergidos nos cultivares de Z. mays variou de 213,17 a 74,0, e o menor número de insetos foi registrado para o cultivar BR 106. Os insetos foram capazes de se alimentar de todos os cultivares, no entanto, o BR 106 mostrou a menor redução na biomassa seca. As lectinas foram detectadas em extratos de BR 106, BRS Asa Branca, BRS Sertanejo e BRS Gorutuba, e a maior atividade foi demonstrada pela BR 106. A menor assimilação de proteína foi detectada nos insetos que se alimentaram com BRS Asa Branca. Os extratos de todos os cultivares foram capazes de inibir a atividade da tripsina bovina, mas esse efeito não está correlacionado ao grau de resistência dos cultivares de Z. mays. Os resultados sugerem a resistência da BR 160 ao ataque de S. zeamais, além de indicar que a presença de lectina nos grãos é a causa dessa resistência.(AU)

The differential expression and snp analysis of the ovoinhibitor gene in the ovaries of laying duck breeds (Anas platyrhynchos)

Ovoinhibitor (OIH) is the main proteinase inhibitor in the egg white. In the present study, real-time quantitative PCR and Western-Blot were used to analyze different expression pattern of OIH in ovaries as a candidate gene of reproductive traits in Jingjiang ducks (JJ ducks) and Shaoxing ducks (SX ducks) during three laying stages. To study the polymorphism of the OIH gene in those two duck populations, we designed five pairs of primers to detect SNPs of exon 3-5, 5-6, 14-16 and intron 7, 9 of the OIH gene by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA pool sequencing methods. The results showed that OIH expression increased during the laying stage in the ovaries of both duck breeds. The relative expression levels of OIH in the egg at hatch and 180days of age were lower in JJ ducks than in SX ducks, but higher in JJ ducks than SX ducks at 500 days of age. Only exon 5-6 locus had a novel SNP. One variation (389G>A) was detected in the two tested duck populations and it was associated with some laying traits, such as body weight of hatch, age at first egg, weight at first egg, egg number at 72weeks of age. The AG genotype was associated with inferior body weight of hatch and superior weight at first egg, age at first egg and egg number at 72weeks of age. Therefore, these results suggest that OIH may be a strong candidate gene related to some laying traits in ducks.(AU)

The differential expression and snp analysis of the ovoinhibitor gene in the ovaries of laying duck breeds (Anas platyrhynchos)

Ovoinhibitor (OIH) is the main proteinase inhibitor in the egg white. In the present study, real-time quantitative PCR and Western-Blot were used to analyze different expression pattern of OIH in ovaries as a candidate gene of reproductive traits in Jingjiang ducks (JJ ducks) and Shaoxing ducks (SX ducks) during three laying stages. To study the polymorphism of the OIH gene in those two duck populations, we designed five pairs of primers to detect SNPs of exon 3-5, 5-6, 14-16 and intron 7, 9 of the OIH gene by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA pool sequencing methods. The results showed that OIH expression increased during the laying stage in the ovaries of both duck breeds. The relative expression levels of OIH in the egg at hatch and 180days of age were lower in JJ ducks than in SX ducks, but higher in JJ ducks than SX ducks at 500 days of age. Only exon 5-6 locus had a novel SNP. One variation (389G>A) was detected in the two tested duck populations and it was associated with some laying traits, such as body weight of hatch, age at first egg, weight at first egg, egg number at 72weeks of age. The AG genotype was associated with inferior body weight of hatch and superior weight at first egg, age at first egg and egg number at 72weeks of age. Therefore, these results suggest that OIH may be a strong candidate gene related to some laying traits in ducks.