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.

Effects of supplementation with combination of polyunsaturated oils in diet of horses in maintenance and during marcha training / Efeito da suplementação com combinação de óleos poliinsaturados na dieta de cavalos em manutenção e durante treino de marcha

This study aimed to evaluate the effects of supplementation with polyunsaturated oil on lipid and mineral blood biomarkers in two different groups of horses. Was used 6 healthy horses in maintenance (MT-g) and 10 healthy horses in marcha gait training (GT-g). The MT-g and the GT-g received 100mL and 300mL/day/animal of combination of polyunsaturated oils, respectively, during 8 weeks. Blood was collected in pre-test and after 4 and 8 weeks, after overnight fasting and it was measured: total cholesterol, HDL cholesterol, LDL cholesterol, triglyceride, NEFA, LPS, glycerol, Ca, P, Mg, and Cl. The supplementation produced significant increase in NEFA and reduction in lipase in both groups. In addition, the LPS content in both groups had a large decrease (~35%) from the pre-test to the 8th week of supplementation. In the MT-g, the LDL cholesterol and glycerol increased significantly after 4 weeks but returned to the pre-test levels after 8 weeks, and the triglyceride content decreased (~62%) after 4 weeks. The total cholesterol concentration in the GT-g increased after 4 weeks (~70%), but after 8 weeks it decreased to levels similar to those observed in the pre-test. The mineral biomarkers increased in the MT-g and GT-g, after 8 weeks. In the MT-g and in the GT-g, Ca decreased. In conclusion, supplementation with a combination of polyunsaturated oils increased NEFA concentration and reduced LPS activity in both groups, unlike the concentrations of the other biomarkers that showed different variations between groups, indicating that both NEFA and LPS the changes in their concentrations during supplementation may be more sensitive and more indicated for these types of evaluation. In addition, some blood mineral concentrations reduced significantly in both groups of horses, and thus, they need to be evaluated in horses under chronic oil supplementation.

O objetivo deste estudo foi avaliar os efeitos da suplementação com uma combinação de óleos poliinsaturado nos biomarcadores lipídicos e minerais sanguíneos em dois diferentes grupos de equinos. Foram utilizados 6 equinos saudáveis em manutenção (MT-g) e 10 equinos saudáveis em treinamento de marcha (GT-g). O MT-g e o GT-g receberam 100,0mL e 300mL/dia/animal de uma combinação de óleos poliinsaturado, respectivamente, durante 8 semanas. O sangue foi coletado no pré-teste e após 4 e 8 semanas, após a noite, medidas: colesterol total, colesterol HDL, colesterol LDL, triglicérides, NEFA, LPS, glicerol, Ca, P, Mg e Cl. A suplementação produziu aumento significativo de NEFA e redução da lipase nos dois grupos. Além disso, o conteúdo de LPS em ambos os grupos ocorreu um decréscimo (~ 35%) desde o pré-teste até a 8ª semana de suplementação. Na MT-g, o colesterol LDL e o glicerol aumentaram significativamente após 4 semanas, mas retornaram aos níveis pré-teste após 8 semanas, e o conteúdo de triglicérides diminuiu (~ 62%) após 4 semanas. A concentração total de colesterol no GT-g aumentou após 4 semanas (~ 70%), mas após 8 semanas diminuiu para valores semelhantes aos observados no pré-teste. Os biomarcadores minerais aumentaram no MT-g e GT-g, após 8 semanas. No MT-g e no GT-g, o Ca diminuiu. Em conclusão, a suplementação com uma combinação de óleos poliinsaturado elevou a concentração do NEFA e reduziu a atividade de LPS em ambos os grupos estudados, diferentemente das concentrações das demais biomarcadores que apresentaram variações diferentes entre os grupos, indicando que tanto o NEFA como a LPS podem ser mais sensíveis as modificações em suas concentrações durante a suplementação e mais indicados para esse tipo de avaliação. Além disso, a concentração sanguínea de alguns minerais reduziu significativamente em ambos os grupos de equinos e, portanto, precisam ser avaliados em cavalos sob suplementação crônica de óleos.

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)

Effects of Lysophospholipid and Lipase Enzyme Supplementation to Low Metabolizable Energy Diets on Growth Performance, Intestinal Morphology and Microbial Population and Some Blood Metabolites in Broiler Chickens

This experiment aimed to evaluate the impacts of dietary lysophospholipid (LPL) and lipase enzyme complementation based on low-energy diets on growth performance, intestinal morphology, blood metabolites, immune response, and carcass traits in broiler chickens. Two hundreds broiler chicks were assigned to a completely randomized design with five treatments and four replications with ten one-day old chicks. The five treatments were: positive control (PC) without LPL supplementation and adequate in all nutrients, negative control (NC) without LPL the reduced 150 kcal/kg of metabolizable energy, NC+ 0.15% LPL (LPL15), NC+ lipase (NCL), NC+ 0.15% LPL+ lipase (NCLL). Feeding LPL improved body weight gain and feed conversion ratio (FCR). In contrast, lipase supplementation showed no significant improvement on weight gain and FCR. Supplementation of LPL and lipase did not have significant effect on immune organ, abdominal fat, and liver and thigh but decreased heart and gizzard and increased breast relative weight (p 0.05). Digestibility of dry matter did not show significant effect but crude protein and ether extract improved digestibility in LPL15 and NCLL group in contrast to NC group (p 0.05). Dietary treatment showed no significant improvement on the metabolic blood factors (p 0.05). The inclusion of LPL to negative diet (LPL15) and LPL+lipase to negative control diet raised villus height, ratio of villi height to crypt depth and increased crypt depth. Overall, LPL inclusion to diet increased weight gain and improved FCR, crude protein and fat digestibility, and improved villus height and ratio of villi height to crypt depth to NC group.(AU)

Effects of supplementation with combination of polyunsaturated oils in diet of horses in maintenance and during marcha training / Efeito da suplementação com combinação de óleos poliinsaturados na dieta de cavalos em manutenção e durante treino de marcha

This study aimed to evaluate the effects of supplementation with polyunsaturated oil on lipid and mineral blood biomarkers in two different groups of horses. Was used 6 healthy horses in maintenance (MT-g) and 10 healthy horses in marcha gait training (GT-g). The MT-g and the GT-g received 100mL and 300mL/day/animal of combination of polyunsaturated oils, respectively, during 8 weeks. Blood was collected in pre-test and after 4 and 8 weeks, after overnight fasting and it was measured: total cholesterol, HDL cholesterol, LDL cholesterol, triglyceride, NEFA, LPS, glycerol, Ca, P, Mg, and Cl. The supplementation produced significant increase in NEFA and reduction in lipase in both groups. In addition, the LPS content in both groups had a large decrease (~35%) from the pre-test to the 8th week of supplementation. In the MT-g, the LDL cholesterol and glycerol increased significantly after 4 weeks but returned to the pre-test levels after 8 weeks, and the triglyceride content decreased (~62%) after 4 weeks. The total cholesterol concentration in the GT-g increased after 4 weeks (~70%), but after 8 weeks it decreased to levels similar to those observed in the pre-test. The mineral biomarkers increased in the MT-g and GT-g, after 8 weeks. In the MT-g and in the GT-g, Ca decreased. In conclusion, supplementation with a combination of polyunsaturated oils increased NEFA concentration and reduced LPS activity in both groups, unlike the concentrations of the other biomarkers that showed different variations between groups, indicating that both NEFA and LPS the changes in their concentrations during supplementation may be more sensitive and more indicated for these types of evaluation. In addition, some blood mineral concentrations reduced significantly in both groups of horses, and thus, they need to be evaluated in horses under chronic oil supplementation.(AU)

O objetivo deste estudo foi avaliar os efeitos da suplementação com uma combinação de óleos poliinsaturado nos biomarcadores lipídicos e minerais sanguíneos em dois diferentes grupos de equinos. Foram utilizados 6 equinos saudáveis em manutenção (MT-g) e 10 equinos saudáveis em treinamento de marcha (GT-g). O MT-g e o GT-g receberam 100,0mL e 300mL/dia/animal de uma combinação de óleos poliinsaturado, respectivamente, durante 8 semanas. O sangue foi coletado no pré-teste e após 4 e 8 semanas, após a noite, medidas: colesterol total, colesterol HDL, colesterol LDL, triglicérides, NEFA, LPS, glicerol, Ca, P, Mg e Cl. A suplementação produziu aumento significativo de NEFA e redução da lipase nos dois grupos. Além disso, o conteúdo de LPS em ambos os grupos ocorreu um decréscimo (~ 35%) desde o pré-teste até a 8ª semana de suplementação. Na MT-g, o colesterol LDL e o glicerol aumentaram significativamente após 4 semanas, mas retornaram aos níveis pré-teste após 8 semanas, e o conteúdo de triglicérides diminuiu (~ 62%) após 4 semanas. A concentração total de colesterol no GT-g aumentou após 4 semanas (~ 70%), mas após 8 semanas diminuiu para valores semelhantes aos observados no pré-teste. Os biomarcadores minerais aumentaram no MT-g e GT-g, após 8 semanas. No MT-g e no GT-g, o Ca diminuiu. Em conclusão, a suplementação com uma combinação de óleos poliinsaturado elevou a concentração do NEFA e reduziu a atividade de LPS em ambos os grupos estudados, diferentemente das concentrações das demais biomarcadores que apresentaram variações diferentes entre os grupos, indicando que tanto o NEFA como a LPS podem ser mais sensíveis as modificações em suas concentrações durante a suplementação e mais indicados para esse tipo de avaliação. Além disso, a concentração sanguínea de alguns minerais reduziu significativamente em ambos os grupos de equinos e, portanto, precisam ser avaliados em cavalos sob suplementação crônica de óleos.(AU)

Effects of Lysophospholipid and Lipase Enzyme Supplementation to Low Metabolizable Energy Diets on Growth Performance, Intestinal Morphology and Microbial Population and Some Blood Metabolites in Broiler Chickens

This experiment aimed to evaluate the impacts of dietary lysophospholipid (LPL) and lipase enzyme complementation based on low-energy diets on growth performance, intestinal morphology, blood metabolites, immune response, and carcass traits in broiler chickens. Two hundreds broiler chicks were assigned to a completely randomized design with five treatments and four replications with ten one-day old chicks. The five treatments were: positive control (PC) without LPL supplementation and adequate in all nutrients, negative control (NC) without LPL the reduced 150 kcal/kg of metabolizable energy, NC+ 0.15% LPL (LPL15), NC+ lipase (NCL), NC+ 0.15% LPL+ lipase (NCLL). Feeding LPL improved body weight gain and feed conversion ratio (FCR). In contrast, lipase supplementation showed no significant improvement on weight gain and FCR. Supplementation of LPL and lipase did not have significant effect on immune organ, abdominal fat, and liver and thigh but decreased heart and gizzard and increased breast relative weight (p 0.05). Digestibility of dry matter did not show significant effect but crude protein and ether extract improved digestibility in LPL15 and NCLL group in contrast to NC group (p 0.05). Dietary treatment showed no significant improvement on the metabolic blood factors (p 0.05). The inclusion of LPL to negative diet (LPL15) and LPL+lipase to negative control diet raised villus height, ratio of villi height to crypt depth and increased crypt depth. Overall, LPL inclusion to diet increased weight gain and improved FCR, crude protein and fat digestibility, and improved villus height and ratio of villi height to crypt depth to NC group.

Nitrogen source and pH interact and modulate lipase secretion in a non-clinical strain of Candida parapsilosis

Lipases (E.C. 3.1.1.3) are serine-hydrolases, and act on long chain fatty acid ester bonds. They exhibit specific and enantioselective activities, which are desirable for many industrial applications. This study aimed at screening and optimizing the production of lipases by wild yeast strains from a variety of substrates, as well as characterizing the enzyme. An initial selection was made in oxygenated oil-supplemented minimum medium, and the enzymatic activity of the supernatant was tested over p-nitrophenyl palmitate. One-hundred and twenty-four yeast strains from different substrates were tested, and twenty-three showed significantly higher lipolytic activity (p<0.01). One yeast in particular, QU110, showed best lipase production and therefore was selected for the optimization and characterization processes. This yeast exhibits enzyme secretion in initial pH 6.0, with olive oil and tryptone as carbon and nitrogen sources, respectively. There was a strong interaction between nitrogen source and initial pH, and pH 9.0seems to inhibit enzyme secretion. The crude enzyme (cell-free supernatant) shows stability in surfactants and n-hexane, but not in ethanol or methanol. A Response Surface Model was created and optimal enzyme activity conditions were observed at 36°C and pH 8.0. The lipase is appropriate for transesterification reactions, as the enzyme is more stable in strong apolar solvents than moderately apolar ones. Also, secretion by pH was not reported elsewhere, which should be further investigated and contribute for other yeast bioprocesses as well.

Use of emulsifier and lipase in feeds for broiler chickens / Uso de emulsificante e lipase em rações de frangos de corte

The objective of this study was to evaluate the effect of exogenous emulsifier and lipase in diets on performance, digestibility, and organ biometry of broiler chickens. A completely randomised design with seven treatments and seven replications was adopted. The treatments were as follows: T1 (positive control; PC): 3000, 3100, 3200, and 3250 Kcal of metabolisable energy (ME) kg-1 of diet for phases 1 to 10, 11 to 21, 22 to 31, and 32 to 37 days, respectively; T2: PC with reduction in ME of 30 Kcal kg-1 of diet; T3: PC with reduction in ME of 60 Kcal kg-1 of feed; T4 (negative control; NC): PC with reduction in ME of 90 Kcal kg-1 of feed; T5: NC with inclusion of exogenous lipase (10 000 U kg-1); T6: NC with inclusion of emulsifier (250 g t-1); and T7: NC with inclusion of lipase (10 000 U kg-1) and emulsifier (250 g t-1) in the period from 1 to 37 days of rearing. Performance characteristics (weight gain (WG), feed intake (FI), and feed conversion (FC)), carcass yield, cut yield, the relative weight of abdominal fat and organs (small intestine, liver, and pancreas), and relative intestinal length, in addition to dry matter digestibility (CDADM), ethereal extract (CDAEE), crude metabolisable energy (CMACE), and apparent metabolisable energy (AME) of the diets, were evaluated. In the initial phase, the CDAEE was higher for the PC group than for the emulsifier + lipase group. The AME determined in the final phase for the group supplemented with an emulsifier was higher by approximately 50 Kcal (EM) than the NC group. The WG of the lipase group was similar to that of the PC group. However, the groups with emulsifier and emulsifier + lipase showed a lower WG than the PC group. The additives used did not recover the FC to the same level observed in the PC group. The reduction in ME of 90 Kcal kg-1 generated a lower WG and worse FC...

Objetivou-se com este estudo avaliar a utilização de emulsificante e lipase em rações sobre a digestibilidade, desempenho e biometria de órgãos para frangos de corte. Foi adotado um delineamento inteiramente casualizado, com sete tratamentos e sete repetições. Os tratamentos foram constituídos da seguinte forma: T1 (Controle Positivo - CP): 3000, 3100, 3200, 3250 Kcal de energia metabolizável (EM) kg-1 de ração para as fases 1 a 10, 11 a 21, 22 a 31 e 32 a 37 dias, respectivamente; T2: CP com redução de EM em 30 Kcal kg-1 de ração; T3: CP com redução de 60 Kcal EM kg-1 de ração; T4 (Controle Negativo - CN): CP com redução de 90 Kcal EM kg-1 de ração; T5: CN + inclusão de lipase exógena (10000 U kg-1); T6: CN com inclusão de emulsificante (250 g t-1); T7: CN com a utilização de lipase (10000 U kg-1) e emulsificante (250 g t-1) no período de 1 a 37 dias de criação. Foram avaliadas características de desempenho (ganho de peso (GP), consumo de ração (CR) e conversão alimentar (CA)), rendimento de carcaça, rendimento de cortes, peso relativo da gordura abdominal e de órgãos (intestino delgado, fígado e pâncreas), comprimento relativo do intestino e digestibilidade da matéria seca (CDAMS), do extrato etéreo (CDAEE) e metabolizabilidade da energia bruta (CMAEB) das rações. Na fase inicial, o CDAEE foram maiores para o grupo CP quando comparado ao grupo com emulsificante+lipase. A EMA determinada na fase final do grupo submetido ao uso de emulsificante apresentou superioridade em cerca de 50 Kcal (EM) em comparação ao grupo CN. O GP do grupo lipase foi semelhante ao observado no grupo CP. Por outro lado, os grupos com emulsificante e emulsificante + lipase apresentaram menor GP quando comparado ao grupo CP. Os aditivos utilizados não proporcionaram recuperação na CA àquela observada no grupo CP. A redução de EMA em 90 Kcal/kg gerou menor GP e pior CA. O uso de aditivos em conjunto foi o grupo que mais aproximou ao desempenho observado no grupo CP...

Use of emulsifier and lipase in feeds for broiler chickens / Uso de emulsificante e lipase em rações de frangos de corte

The objective of this study was to evaluate the effect of exogenous emulsifier and lipase in diets on performance, digestibility, and organ biometry of broiler chickens. A completely randomised design with seven treatments and seven replications was adopted. The treatments were as follows: T1 (positive control; PC): 3000, 3100, 3200, and 3250 Kcal of metabolisable energy (ME) kg-1 of diet for phases 1 to 10, 11 to 21, 22 to 31, and 32 to 37 days, respectively; T2: PC with reduction in ME of 30 Kcal kg-1 of diet; T3: PC with reduction in ME of 60 Kcal kg-1 of feed; T4 (negative control; NC): PC with reduction in ME of 90 Kcal kg-1 of feed; T5: NC with inclusion of exogenous lipase (10 000 U kg-1); T6: NC with inclusion of emulsifier (250 g t-1); and T7: NC with inclusion of lipase (10 000 U kg-1) and emulsifier (250 g t-1) in the period from 1 to 37 days of rearing. Performance characteristics (weight gain (WG), feed intake (FI), and feed conversion (FC)), carcass yield, cut yield, the relative weight of abdominal fat and organs (small intestine, liver, and pancreas), and relative intestinal length, in addition to dry matter digestibility (CDADM), ethereal extract (CDAEE), crude metabolisable energy (CMACE), and apparent metabolisable energy (AME) of the diets, were evaluated. In the initial phase, the CDAEE was higher for the PC group than for the emulsifier + lipase group. The AME determined in the final phase for the group supplemented with an emulsifier was higher by approximately 50 Kcal (EM) than the NC group. The WG of the lipase group was similar to that of the PC group. However, the groups with emulsifier and emulsifier + lipase showed a lower WG than the PC group. The additives used did not recover the FC to the same level observed in the PC group. The reduction in ME of 90 Kcal kg-1 generated a lower WG and worse FC...(AU)

Objetivou-se com este estudo avaliar a utilização de emulsificante e lipase em rações sobre a digestibilidade, desempenho e biometria de órgãos para frangos de corte. Foi adotado um delineamento inteiramente casualizado, com sete tratamentos e sete repetições. Os tratamentos foram constituídos da seguinte forma: T1 (Controle Positivo - CP): 3000, 3100, 3200, 3250 Kcal de energia metabolizável (EM) kg-1 de ração para as fases 1 a 10, 11 a 21, 22 a 31 e 32 a 37 dias, respectivamente; T2: CP com redução de EM em 30 Kcal kg-1 de ração; T3: CP com redução de 60 Kcal EM kg-1 de ração; T4 (Controle Negativo - CN): CP com redução de 90 Kcal EM kg-1 de ração; T5: CN + inclusão de lipase exógena (10000 U kg-1); T6: CN com inclusão de emulsificante (250 g t-1); T7: CN com a utilização de lipase (10000 U kg-1) e emulsificante (250 g t-1) no período de 1 a 37 dias de criação. Foram avaliadas características de desempenho (ganho de peso (GP), consumo de ração (CR) e conversão alimentar (CA)), rendimento de carcaça, rendimento de cortes, peso relativo da gordura abdominal e de órgãos (intestino delgado, fígado e pâncreas), comprimento relativo do intestino e digestibilidade da matéria seca (CDAMS), do extrato etéreo (CDAEE) e metabolizabilidade da energia bruta (CMAEB) das rações. Na fase inicial, o CDAEE foram maiores para o grupo CP quando comparado ao grupo com emulsificante+lipase. A EMA determinada na fase final do grupo submetido ao uso de emulsificante apresentou superioridade em cerca de 50 Kcal (EM) em comparação ao grupo CN. O GP do grupo lipase foi semelhante ao observado no grupo CP. Por outro lado, os grupos com emulsificante e emulsificante + lipase apresentaram menor GP quando comparado ao grupo CP. Os aditivos utilizados não proporcionaram recuperação na CA àquela observada no grupo CP. A redução de EMA em 90 Kcal/kg gerou menor GP e pior CA. O uso de aditivos em conjunto foi o grupo que mais aproximou ao desempenho observado no grupo CP...(AU)

Enzymatic comparison and mortality of Beauveria bassiana against cabbage caterpillar Pieris brassicae LINN

Beauveria bassiana, an entomopathogenic fungus, is the alternative biocontrol agent exploited against major economic crop pests. Pieris brassicae L. is an emerging pest of the Brassicaceae family. Therefore, in the present study, fungal isolates of Beauveria bassiana, viz. MTCC 2028, MTCC 4495, MTCC 6291, and NBAII-11, were evaluated for their virulence against third instar larvae of P. brassicae. Among all these fungal isolates, maximum mortality (86.66%) was recorded in B. bassiana MTCC 4495 at higher concentration of spores (109 conidia/ml), and the minimum mortality (30.00%) was recorded in B. bassiana MTCC 6291 at a lower concentration (107 conidia/ml) after ten days of treatment. The extracellular cuticle-degrading enzyme activities of fungal isolates were measured. Variability was observed both in the pattern of enzyme secretion and the level of enzyme activities among various fungal isolates. B. bassiana MTCC 4495 recorded the maximum mean chitinase (0.51 U/ml), protease (1.12 U/ml), and lipase activities (1.36 U/ml). The minimum mean chitinase and protease activities (0.37 and 0.91 U/ml, respectively) were recorded in B. bassiana MTCC 6291. The minimum mean lipase activity (1.04 U/ml) was recorded in B. bassiana NBAII-11. Our studies revealed B. bassiana MTCC 4495 as the most pathogenic isolate against P. brassicae, which also recorded maximum extracellular enzyme activities, suggesting the possible roles of extracellular enzymes in the pathogenicity of B. bassiana against P. brassicae.(AU)

Characterization of biotechnologically relevant extracellular lipase produced by Aspergillus terreus NCFT 4269.10

Enzyme production by Aspergillus terreus NCFT 4269.10 was studied under liquid static surface and solid-state fermentation using mustard oil cake as a substrate. The maximum lipase biosynthesis was observed after incubation at 30 °C for 96 h. Among the domestic oils tested, the maximum lipase biosynthesis was achieved using palm oil. The crude lipase was purified 2.56-fold to electrophoretic homogeneity, with a yield of 8.44%, and the protein had a molecular weight of 46.3 kDa as determined by SDS-PAGE. Enzyme characterization confirmed that the purified lipase was most active at pH 6.0, temperature of 50 °C, and substrate concentration of 1.5%. The enzyme was thermostable at 60 °C for 1 h, and the optimum enzymesubstrate reaction time was 30 min. Sodium dodecyl sulfate and commercial detergents did not significantly affect lipase activity during 30-min incubation at 30 °C. Among the metal ions tested, the maximum lipase activity was attained in the presence of Zn2+, followed by Mg2+ and Fe2+. Lipase activity was not significantly affected in the presence of ethylenediaminetetraacetic acid, sodium lauryl sulfate and Triton X-100. Phenylmethylsulfonyl fluoride (1 mM) and the reducing, -mercaptoethanol significantly inhibited lipase activity. The remarkable stability in the presence of detergents, additives, inhibitors and metal ions makes this lipase unique and a potential candidate for significant biotechnological exploitation. (AU)

Surface response methodology for the optimization of lipase production under submerged fermentation by filamentous fungi

A PlackettBurman Factorial Design of 16 experiments was conducted to assess the influence of nine factors on the production of lipases by filamentous fungi. The factors investigated were bran type (used as the main carbon source), nitrogen source, nitrogen source concentration, inducer, inducer concentration, fungal strain (Aspergillus niger or Aspergillus flavus were selected as good lipase producers via submerged fermentation), pH and agitation. The concentration of the yeast extract and soybean oil and the pH had a significant effect (p 0.05) on lipase production and were consecutively studied through a Full Factorial Design 23, with the concentration of yeast extract and pH being significant (p 0.05). These variables were optimized using a central composite design, obtaining maximum lipolytic activities with the use of 45 g/L of yeast extract and pH 7.15. The statistical model showed a 94.12% correlation with the experimental data.(AU)

Process optimization for production and purification of a thermostable, organic solvent tolerant lipase from Acinetobacter sp. AU07

The purpose of this study was to isolate, purify and optimize the production conditions of an organic solvent tolerant and thermostable lipase from Acinetobacter sp. AU07 isolated from distillery waste. The lipase production was optimized by response surface methodology, and a maximum production of 14.5 U/mL was observed at 30 ºC and pH 7, using a 0.5% (v/v) inoculum, 2% (v/v) castor oil (inducer), and agitation 150 rpm. The optimized conditions from the shake flask experiments were validated in a 3 L lab scale bioreactor, and the lipase production increased to 48 U/mL. The enzyme was purified by ammonium sulfate precipitation and ion exchange chromatography and the overall yield was 36%. SDS-PAGE indicated a molecular weight of 45 kDa for the purified protein, and Matrix assisted laser desorption/ionization time of flight analysis of the purified lipase showed sequence similarity with GDSL family of lipases. The optimum temperature and pH for activity of the enzyme was found to be 50 ºC and 8.0, respectively. The lipase was completely inhibited by phenylmethylsulfonyl fluoride but minimal inhibition was observed when incubated with ethylenediaminetetraacetic acid and dithiothreitol. The enzyme was stable in the presence of non-polar hydrophobic solvents. Detergents like SDS inhibited enzyme activity; however, there was minimal loss of enzyme activity when incubated with hydrogen peroxide, Tween 80 and Triton X-100. The kinetic constants (Km and Vmax) revealed that the hydrolytic activity of the lipase was specific to moderate chain fatty acid esters. The Vmax, Km and Vmax/Km ratio of the enzyme were 16.98 U/mg, 0.51 mM, and 33.29, respectively when 4-nitrophenyl palmitate was used as a substrate.(AU)

Aproveitamento de resíduos da agroindústria do óleo de dendê para a produção de lipase por Aspergillus Níger / Agro-industrial residues utilization of palm oil for lipase production by Aspergillus niger

Neste trabalho, foi avaliado o desempenho de duas linhagens de Aspergillus niger (mutante 11T53A14 e selvagem C) previamente selecionadas como promissoras para a produção de lipases, utilizando meios de cultivo formulados a partir da torta de dendê (palmiste) e da borra alcalina do refino do óleo de dendê (borra de dendê), resíduos provenientes da agroindústria do óleo de dendê (palma) por fermentação em estado sólido (FES). Os experimentos de produção da enzima em FES foram conduzidos em colunas aeradas, incubadas em banho-maria a 32ºC com entrada controlada de ar não umedecido de 1,0vvm. Os melhores resultados foram obtidos com a cepa mutante A. niger 11T53A14 em meio contendo torta de dendê umedecida com solução sulfato de amônio (1,2%) e com a adição de 3% da borra de dendê. O valor máximo da atividade da lipase neste meio foi de 72,57U gss-1 em 48 horas. Esse valor foi 47,5% superior ao obtido no meio sem a borra de dendê. A comparação do desempenho da cepa mutante com a cepa selvagem mostrou que o meio composto por torta de dendê adicionado com sulfato de amônio e borra de dendê induziu ambas as linhagens a produzir lipases com bons níveis de atividade, além de reduzir o tempo de processo de fermentação.(AU)

In this paper it was evaluated the performance of two strains of Aspergillus niger (mutant 11T53A14 and wild type C) previously selected as promising for lipase production, from cultivation media formulated from palm kernel cake (kernel) and alkaline sludge from refining were evaluated palm oil (palm oil sludge), palm oil (palm) waste industrialization by solid state fermentation (SSF). Experiments of enzyme production in SSF were conducted in aerated columns, incubated in a water bath at 32°C with controlled inlet of 1.0vvm. The best results were obtained with the mutant strain A. niger 11T53A14 in medium containing palm kernel cake moistened with a solution of ammonium sulfate (1.2%) and with the addition of 3% of palm oil sludge. The maximum lipase activity in this medium was 72.57U gdw-1 in 48 hours and 47.5% higher than in the medium without sludge palm. A comparison of the performance of the mutant strain with the wild-type strain showed that the medium composed of palm kernel cake added with ammonium sulfate and blurs palm induced both strains to produce lipases with good activity levels and reduced the time of the fermentation process.(AU)

Synthesis of structured triacylglycerols enriched in n-3 fatty acids by immobilized microbial lipase

The search for new biocatalysts has aroused great interest due to the variety of micro-organisms and their role as enzyme producers. Native lipases from Aspergillus niger and Rhizopus javanicus were used to enrich the n-3 long-chain polyunsaturated fatty acids content in the triacylglycerols of soybean oil by acidolysis with free fatty acids from sardine oil in solvent-free media. For the immobilization process, the best lipase/support ratios were 1:3 (w/w) for Aspergillus niger lipase and 1:5 (w/w) for Rhizopus javanicus lipase using Amberlite MB-1. Both lipases maintained constant activity for 6 months at 4 °C. Reaction time, sardine-free fatty acids:soybean oil mole ratio and initial water content of the lipase were investigated to determine their effects on n-3 long-chain polyunsaturated fatty acids incorporation into soybean oil. Structured triacylglycerols with 11.7 and 7.2% of eicosapentaenoic acid + docosahexaenoic acid were obtained using Aspergillus niger lipase and Rhizopus javanicus lipase, decreasing the n-6/n-3 fatty acids ratio of soybean oil (11:1 to 3.5:1 and 4.7:1, respectively). The best reaction conditions were: initial water content of lipase of 0.86% (w/w), sardine-free faty acids:soybean oil mole ratio of 3:1 and reaction time of 36 h, at 40 °C. The significant factors for the acidolysis reaction were the sardine-free fatty acids:soybean oil mole ratio and reaction time. The characterization of structured triacylglycerols was obtained using easy ambient sonic-spray ionization mass spectrometry. The enzymatic reaction led to the formation of many structured triacylglycerols containing eicosapentaenoic acid, docosahexaenoic acid or both polyunsaturated fatty acids.(AU)

Assessment of lipolytic activity of isolated microorganisms from the savannah of the Tocantins / Avaliação da atividade lipolítica de microrganismos isolados do Cerrado tocantinense

Current study assesses the biodiversity and selects lipase-producer microorganisms with industrial interest, from the savannah of the state of Tocantins, Brazil. Seventeen pequi microorganisms (Caryocar brasiliense) were isolated in the decomposition stage and 35 microorganisms were retrieved from the soil fraction under the collected pequi. Yarrowia lypolitica strain was used as positive control in all assays. The 52 strains were subjected to tests in a solid medium with Tween 20 for checking halos formed by crystals, indicating lipase production by inoculated strains. Another test to confirm lipase producers was conducted in microplates with liquid medium and enriched with p-nitrophenyl palmitate (pNPP) monitored at 410 m. The AS16 and AP5 strains showed the highest activity for test conditions, namely, 0.072 and 0.067 U mL-1 respectively. Rates were higher than the lipase activity of Yarrowia lypolitica(0.052 U mL-1), a reference strain in current assay.(AU)

O atual estudo avalia a biodiversidade e selecionou microorganismos produtores de lipases de interesse industrial, a partir do cerrado tocantinense. O principal objetivo deste trabalho é avaliar a biodiversidade e selecionar microrganismos do cerrado tocantinense, produtores de lipases de interesse industrial. Foram isolados 17 microrganismos de Pequi (Caryocar brasiliense) em estágio de decomposição e também 35 microrganismos provenientes da fração do solo que se encontrava abaixo do pequi coletado. Foi utilizada uma cepa de Yarrowia lypolitica como controle positivo em todos os ensaios. Todas as 52 cepas foram submetidas a testes em meio sólido, utilizando Tween 20 para a verificação da presença de halos formados por cristais, indicando produção de lipase pelas linhagens inoculadas. Outro teste para confirmação dos produtores de lipase foi realizado em microplacas contendo meio líquido e enriquecido com pNPP (Palmitato de p-nitrofenila), monitorado a 410 m. As cepas AS16 e AP5 apresentaram as maiores atividades para as condições de ensaio a que foram submetidas: 0,072 e 0,067 U mL-1 respectivamente, sendo estes valores maiores que a atividade lipolítica da Yarrowia lypolitica (0,052 U mL-1), considerada como cepa referência para este trabalho.(AU)

Gene cloning, expression, and characterization of the Bacillus amyloliquefaciens PS35 lipase

Abstract Lipases are enzymes of immense industrial relevance, and, therefore, are being intensely investigated. In an attempt to characterize lipases at molecular level from novel sources, a lipase gene from Bacillus amyloliquefaciens PS35 was cloned, heterologously expressed in Escherichia coli DH5α cells and sequenced. It showed up to 98% homology with other lipase sequences in the NCBI database. The recombinant enzyme was then purified from E. coli culture, resulting in a 19.41-fold purification with 9.7% yield. It displayed a preference for long-chain para-nitrophenyl esters, a characteristic that is typical of true lipases. Its optimum pH and temperature were determined to be 8.0 and 40 °C, respectively. The half-lives were 2.0, 1.0 and 0.5 h at 50 °C, 60 °C and 70 °C, respectively. The metal ions K+ and Fe3+ enhanced the enzyme activity. The enzyme displayed substantial residual activity in the presence of various tested chemical modifiers, and interestingly, the organic solvents, such as n-hexane and toluene, also favored the enzyme activity. Thus, this study involves characterization of B. amyloliquefaciens lipase at molecular level. The key outcomes are novelty of the bacterial source and purification of the enzyme with desirable properties for industrial applications.(AU)

Milk-deteriorating exoenzymes from Pseudomonas fluorescens 041 isolated from refrigerated raw milk

The practice of refrigerating raw milk at the farm has provided a selective advantage for psychrotrophic bacteria that produce heat-stable proteases and lipases causing severe quality problems to the dairy industry. In this work, a protease (AprX) and a lipase (LipM) produced by Pseudomonas fluorescens 041, a highly proteolytic and lipolytic strain isolated from raw milk obtained from a Brazilian farm, have been purified and characterized. Both enzymes were purified as recombinant proteins from Escherichia coli. The AprX metalloprotease exhibited activity in a broad temperature range, including refrigeration, with a maximum activity at 37 °C. It was active in a pH range of 4.0 to 9.0. This protease had maximum activity with the substrates casein and gelatin in the presence of Ca+2. The LipM lipase had a maximum activity at 25 °C and a broad pH optimum ranging from 7.0 to 10. It exhibited the highest activity, in the presence of Ca+2, on substrates with long-chain fatty acid residues. These results confirm the spoilage potential of strain 041 in milk due to, at least in part, these two enzymes. The work highlights the importance of studies of this kind with strains isolated in Brazil, which has a recent history on the implementation of the cold chain at the dairy farm.(AU)

Concentration, characterization and application of lipases from Sporidiobolus pararoseus strain

Lipases produced by a newly isolated Sporidiobolus pararoseus strain have potential catalytic ability for esterification reactions. After production, the enzymatic extracts (conventional crude and precipitated, 'CC' and 'CP', and industrial crude and precipitated, 'IC' e 'IP') were partially characterized. The enzymes presented, in general, higher specificity for short chain alcohols and fatty acids. The precipitated extract showed a good thermal stability, higher than that for crude enzymatic extracts. The 'CC' and 'CP' enzymes presented high activities after exposure to pH 6.5 and 40 ºC. On the other hand, the 'IC' and 'IP' extracts kept their activities in a wide range of pH memory but presented preference for higher reaction temperatures. Preliminary studies of application of the crude lipase extract in the enzymatic production of geranyl propionate using geraniol and propionic acid as substrates in solvent-free system led to a reaction conversion of 42 ± 1.5%.(AU)

Molecular characterization of a proteolysis-resistant lipase from Bacillus pumilus SG2

Proteolysis-resistant lipases can be well exploited by industrial processes which employ both lipase and protease as biocatalysts. A proteolysis resistant lipase from Bacillus pumilus SG2 was isolated, purified and characterized earlier. The lipase was resistant to native and commercial proteases. In the present work, we have characterized the lip gene which encodes the proteolysis-resistant lipase from Bacillus pumilus SG2. The parameters and structural details of lipase were analysed. The lip gene consisted of 650 bp. The experimental molecular weight of SG2 lipase was nearly double that of its theoretical molecular weight, thus suggesting the existence of the functional lipase as a covalent dimer. The proteolytic cleavage sites of the lipase would have been made inaccessible by dimerisation, thus rendering the lipase resistant to protease.