Carangoides bartholomaei (Cuvier, 1833) stomach: a source of aspartic proteases for industrial and biotechnological applications / Estômago de Carangoides bartholomaei (Cuvier, 1833): uma fonte de proteases aspárticas para aplicações industriais e biotecnológicas

The viscera and other residues from fish processing are commonly discarded by the fishing industry. These by products can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 U·mg-¹. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.

As vísceras e outros resíduos do processamento de peixes são geralmente descartados pela indústria pesqueira. Esses resíduos podem ser uma fonte de enzimas digestivas com potencial industrial e biotecnológico. Neste estudo, objetivamos a extração, caracterização e aplicação de proteases aspárticas do estômago de Carangoides bartholomaei (Cuvier, 1833). Um extrato bruto do estômago foi obtido e submetido a um processo de purificação parcial, que obteve um Extrato Purificado (EP) com uma atividade proteolítica específica de 54,0 U·mg-¹. Foi obtida uma purificação de 1,9 vezes e um rendimento de 41%. O EP apresenta duas isoformas de proteases ácidas e atividade proteolítica máxima a 45 °C e pH 2,0. A atividade proteolítica do EP foi estável na faixa de pH de 1,5 a 7,0 e temperatura de 25 °C a 50 °C. O EP manteve 35% de sua atividade proteolítica na presença de NaCl a 15% (m/v), mas foi totalmente inibida pela pepstatina A. As proteases ácidas do EP apresentaram alta atividade na presença de metais pesados como o Cd2+, Hg2+, Pb2+, Al3+ e Cu2+. A utilização de EP como aditivo enzimático na extração de colágeno a partir de escamas de tilápia do Nilo dobrou o rendimento do processo. Os resultados indicam um potencial dessas proteases para aplicações industriais e biotecnológicas.

Carangoides bartholomaei (Cuvier, 1833) stomach: a source of aspartic proteases for industrial and biotechnological applications / Estômago de Carangoides bartholomaei (Cuvier, 1833): uma fonte de proteases aspárticas para aplicações industriais e biotecnológicas

The viscera and other residues from fish processing are commonly discarded by the fishing industry. These by products can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 U·mg-¹. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.(AU)

As vísceras e outros resíduos do processamento de peixes são geralmente descartados pela indústria pesqueira. Esses resíduos podem ser uma fonte de enzimas digestivas com potencial industrial e biotecnológico. Neste estudo, objetivamos a extração, caracterização e aplicação de proteases aspárticas do estômago de Carangoides bartholomaei (Cuvier, 1833). Um extrato bruto do estômago foi obtido e submetido a um processo de purificação parcial, que obteve um Extrato Purificado (EP) com uma atividade proteolítica específica de 54,0 U·mg-¹. Foi obtida uma purificação de 1,9 vezes e um rendimento de 41%. O EP apresenta duas isoformas de proteases ácidas e atividade proteolítica máxima a 45 °C e pH 2,0. A atividade proteolítica do EP foi estável na faixa de pH de 1,5 a 7,0 e temperatura de 25 °C a 50 °C. O EP manteve 35% de sua atividade proteolítica na presença de NaCl a 15% (m/v), mas foi totalmente inibida pela pepstatina A. As proteases ácidas do EP apresentaram alta atividade na presença de metais pesados como o Cd2+, Hg2+, Pb2+, Al3+ e Cu2+. A utilização de EP como aditivo enzimático na extração de colágeno a partir de escamas de tilápia do Nilo dobrou o rendimento do processo. Os resultados indicam um potencial dessas proteases para aplicações industriais e biotecnológicas.(AU)

Carangoides bartholomaei (Cuvier, 1833) stomach: a source of aspartic proteases for industrial and biotechnological applications

Abstract The viscera and other residues from fish processing are commonly discarded by the fishing industry. These by-products can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 Umg-1. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.

Resumo As vísceras e outros resíduos do processamento de peixes são geralmente descartados pela indústria pesqueira. Esses resíduos podem ser uma fonte de enzimas digestivas com potencial industrial e biotecnológico. Neste estudo, objetivamos a extração, caracterização e aplicação de proteases aspárticas do estômago de Carangoides bartholomaei (Cuvier, 1833). Um extrato bruto do estômago foi obtido e submetido a um processo de purificação parcial, que obteve um Extrato Purificado (EP) com uma atividade proteolítica específica de 54,0 Umg-1. Foi obtida uma purificação de 1,9 vezes e um rendimento de 41%. O EP apresenta duas isoformas de proteases ácidas e atividade proteolítica máxima a 45 °C e pH 2,0. A atividade proteolítica do EP foi estável na faixa de pH de 1,5 a 7,0 e temperatura de 25 °C a 50 °C. O EP manteve 35% de sua atividade proteolítica na presença de NaCl a 15% (m/v), mas foi totalmente inibida pela pepstatina A. As proteases ácidas do EP apresentaram alta atividade na presença de metais pesados como o Cd2+, Hg2+, Pb2+, Al3+ e Cu2+. A utilização de EP como aditivo enzimático na extração de colágeno a partir de escamas de tilápia do Nilo dobrou o rendimento do processo. Os resultados indicam um potencial dessas proteases para aplicações industriais e biotecnológicas

Carangoides bartholomaei (Cuvier, 1833) stomach: a source of aspartic proteases for industrial and biotechnological applications / Estômago de Carangoides bartholomaei (Cuvier, 1833): uma fonte de proteases aspárticas para aplicações industriais e biotecnológicas

The viscera and other residues from fish processing are commonly discarded by the fishing industry. These byproducts can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 U·mg-1. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.

As vísceras e outros resíduos do processamento de peixes são geralmente descartados pela indústria pesqueira. Esses resíduos podem ser uma fonte de enzimas digestivas com potencial industrial e biotecnológico. Neste estudo, objetivamos a extração, caracterização e aplicação de proteases aspárticas do estômago de Carangoides bartholomaei (Cuvier, 1833). Um extrato bruto do estômago foi obtido e submetido a um processo de purificação parcial, que obteve um Extrato Purificado (EP) com uma atividade proteolítica específica de 54,0 U·mg-1. Foi obtida uma purificação de 1,9 vezes e um rendimento de 41%. O EP apresenta duas isoformas de proteases ácidas e atividade proteolítica máxima a 45 °C e pH 2,0. A atividade proteolítica do EP foi estável na faixa de pH de 1,5 a 7,0 e temperatura de 25 °C a 50 °C. O EP manteve 35% de sua atividade proteolítica na presença de NaCl a 15% (m/v), mas foi totalmente inibida pela pepstatina A. As proteases ácidas do EP apresentaram alta atividade na presença de metais pesados como o Cd2+, Hg2+, Pb2+, Al3+ e Cu2+. A utilização de EP como aditivo enzimático na extração de colágeno a partir de escamas de tilápia do Nilo dobrou o rendimento do processo. Os resultados indicam um potencial dessas proteases para aplicações industriais e biotecnológicas.

Influence of foliar-applied glycinebetaine on growth, yield and some key physiological and biochemical attributes of grape (Vitis vinifera L) / Influência da glicinabetaína aplicada via foliar no crescimento, rendimento e alguns atributos fisiológicos e bioquímicos essenciais da uva (Vitis vinifera L)

Glycinebetaine (GB) plays an imperative role to mitigate the opposing impact of several environmental stresses in various crops. The objective of this investigation was to scrutinize the response of grape (Vitis vinifera L.) to the foliar application of GB. Diverse levels of GB (0, 10, 20 and 30 mM) were applied three times with fifteen days interval to grape cv. King's Ruby. All levels of GB improved the physiological, biochemical and growth attributes of the grape. As compared to control treatment, foliar spray of GB applied at 30 mM increased the number of leaves/vine, leaf area and number of newly emerged branches/plant by 20.55%, 12.28% and 48.13%, respectively. The pH of grape juice was decreased by increasing GB levels. However, total soluble solids, total chlorophyll contents, grape yield and photosynthesis rate was recorded maximum with foliar spray of GB applied at 30 mM. The regression model predicted that each increment in GB level enhanced the number of leaves/plant and the number of newly emerged branches/plant by 1.8 and 0.7, respectively. Thus, it is concluded that exogenous application GB applied at 30 mM might be more useful to obtain the optimum growth and quality of grapes.

A glicina betaína (GB) desempenha um papel fundamental para mitigar o impacto oposto de vários estresses ambientais em várias culturas agrícolas. O objetivo desta pesquisa foi examinar a resposta da uva (Vitis vinifera L.) à aplicação foliar de GB. Diversos níveis de GB (0, 10, 20 e 30 mM) foram aplicados três vezes com intervalo de 15 dias à videira cv. King's Ruby. Todos os níveis de GB melhoraram os atributos fisiológicos, bioquímicos e de crescimento da uva. Em comparação ao tratamento controle, a pulverização foliar de GB aplicada à 30 mM aumentou o número de folhas / videira, área foliar e número de ramos / planta recém-emergidos em 20,55%, 12,28% e 48,13%, respectivamente. O pH do suco de uva diminuiu com o aumento dos níveis de GB. No entanto, sólidos solúveis totais, conteúdo de clorofila total, rendimento de uva e taxa de fotossíntese foram registrados no máximo com pulverização foliar de GB aplicada a 30 mM. O modelo de regressão previu que cada incremento no nível de GB aumentava o número de folhas/planta e o número de ramos/planta recém-emergidos em 1,8 e 0,7, respectivamente. Assim, conclui-se que a aplicação exógena GB aplicada à 30 mM pode ser mais útil para obter o crescimento e a qualidade ideais das uvas.

Influence of foliar-applied glycinebetaine on growth, yield and some key physiological and biochemical attributes of grape (Vitis vinifera L) / Influência da glicinabetaína aplicada via foliar no crescimento, rendimento e alguns atributos fisiológicos e bioquímicos essenciais da uva (Vitis vinifera L)

Glycinebetaine (GB) plays an imperative role to mitigate the opposing impact of several environmental stresses in various crops. The objective of this investigation was to scrutinize the response of grape (Vitis vinifera L.) to the foliar application of GB. Diverse levels of GB (0, 10, 20 and 30 mM) were applied three times with fifteen days interval to grape cv. King's Ruby. All levels of GB improved the physiological, biochemical and growth attributes of the grape. As compared to control treatment, foliar spray of GB applied at 30 mM increased the number of leaves/vine, leaf area and number of newly emerged branches/plant by 20.55%, 12.28% and 48.13%, respectively. The pH of grape juice was decreased by increasing GB levels. However, total soluble solids, total chlorophyll contents, grape yield and photosynthesis rate was recorded maximum with foliar spray of GB applied at 30 mM. The regression model predicted that each increment in GB level enhanced the number of leaves/plant and the number of newly emerged branches/plant by 1.8 and 0.7, respectively. Thus, it is concluded that exogenous application GB applied at 30 mM might be more useful to obtain the optimum growth and quality of grapes.(AU)

A glicina betaína (GB) desempenha um papel fundamental para mitigar o impacto oposto de vários estresses ambientais em várias culturas agrícolas. O objetivo desta pesquisa foi examinar a resposta da uva (Vitis vinifera L.) à aplicação foliar de GB. Diversos níveis de GB (0, 10, 20 e 30 mM) foram aplicados três vezes com intervalo de 15 dias à videira cv. King's Ruby. Todos os níveis de GB melhoraram os atributos fisiológicos, bioquímicos e de crescimento da uva. Em comparação ao tratamento controle, a pulverização foliar de GB aplicada à 30 mM aumentou o número de folhas / videira, área foliar e número de ramos / planta recém-emergidos em 20,55%, 12,28% e 48,13%, respectivamente. O pH do suco de uva diminuiu com o aumento dos níveis de GB. No entanto, sólidos solúveis totais, conteúdo de clorofila total, rendimento de uva e taxa de fotossíntese foram registrados no máximo com pulverização foliar de GB aplicada a 30 mM. O modelo de regressão previu que cada incremento no nível de GB aumentava o número de folhas/planta e o número de ramos/planta recém-emergidos em 1,8 e 0,7, respectivamente. Assim, conclui-se que a aplicação exógena GB aplicada à 30 mM pode ser mais útil para obter o crescimento e a qualidade ideais das uvas.(AU)

Respostas da aplicação foliar de glicina betaína em cana-de-açúcar submetida a estresse hídrico e reidratação / Responses of the leaf application of glycine betaine on sugarcane submitted to drought stress and rehydration

O crescimento das plantas de cana-de-açúcar é bastante reduzido devido à ocorrência de déficit hídrico, o que acaba afetando a produção. Diante disso, vem sendo utilizadas diferentes estratégias para atenuar condições de estresse, como a aplicação exógena de glicina betaína. Com este trabalho, objetivou-se avaliar a taxa de crescimento e a massa seca de cana-de-açúcar aplicada glicina betaína, e submetida adéficit hídrico e reidratação. O experimento foi realizado em casa de vegetação com seis tratamentos: G0I - plantas sem glicina betaína com irrigação adequada, G0E - plantas sem glicina betaína e estresse hídrico, G1I - plantas com uma aplicação de glicina betaína e irrigação adequada, G1E - plantas com umaaplicação de glicina betaína e estresse hídrico, G2I - plantas com duas aplicações de glicina betaína e irrigação adequada, G2E - plantas com duas aplicações de glicina betaína e estresse hídrico. Aos seis dias após o estresse as plantas foram reidratadas. As taxas relativas de crescimento das plantas de cana-de-açúcar reduziram com o estresse hídrico. No entanto, estas reduções foram minimizadas com a aplicação foliar de glicina betaína, principalmente, com a reidratação em que a taxa relativa de crescimento reduziu apenas 19 e 18,4%, respectivamente, em G1 e G2, enquanto em G0 a redução foi de 37,2%. A glicina betaína também minimizou as reduções na produção de massa seca total sob estresse hídrico, com reduções de 5,8 e 5,9% em G1 e G2, respectivamente, enquanto em G0 foi de 15,1%. A aplicação foliar de glicina betaína melhora a taxa relativa de crescimento de plantas de cana-de-açúcar sob estresse hídrico, sobretudo após a reidratação o que diminui os efeitos negativos do estresse sobre a produção de massa seca.

The growth of sugarcane plants is greatly reduced due to the occurrence of drought stress, which ultimately affects production. Therefore, different strategies have been used to attenuate stress conditions, such as the exogenous application of glycine betaine. The objective of this work was to evaluate the growth rate and dry mass of sugarcane that received a glycine betaine application and was submitted todrought stress and rehydration. The experiment was carried out in a greenhouse with six treatments: G0I - plants without glycine betaine with adequate irrigation, G0E - plants without glycine betaine and drought stress, G1I - plants with an application of glycine betaine and adequate irrigation, G1E - plants with an application of glycine betaine and drought stress, G2I - plants with two applications of glycine betaine and adequate irrigation, G2E - plants with two applications of glycine betaine and drought stress. Six days after the stress, the plants were rehydrated. The relative growth rates of sugarcane plants decreased with the drought stress. However, these reductions were partially minimized with the foliar application of glycine betaine, mainly with the rehydration, with the relative growth rate decreasing by only 19% and 18.4%, respectively, in G1 and G2, while the reduction was 37.2% in G0. Glycine betaine also minimized reductions in total dry mass production under drought stress, with decreases of 5.8% and 5.9% in G1 and G2, respectively, while it was 15.1% in G0. The leaf application of glycine betaine improves the relative growth rate of sugarcane plants under drought stress, especially after rehydration, which decreases the negative effects of stress on dry mass production.

Respostas da aplicação foliar de glicina betaína em cana-de-açúcar submetida a estresse hídrico e reidratação / Responses of the leaf application of glycine betaine on sugarcane submitted to drought stress and rehydration

O crescimento das plantas de cana-de-açúcar é bastante reduzido devido à ocorrência de déficit hídrico, o que acaba afetando a produção. Diante disso, vem sendo utilizadas diferentes estratégias para atenuar condições de estresse, como a aplicação exógena de glicina betaína. Com este trabalho, objetivou-se avaliar a taxa de crescimento e a massa seca de cana-de-açúcar aplicada glicina betaína, e submetida adéficit hídrico e reidratação. O experimento foi realizado em casa de vegetação com seis tratamentos: G0I - plantas sem glicina betaína com irrigação adequada, G0E - plantas sem glicina betaína e estresse hídrico, G1I - plantas com uma aplicação de glicina betaína e irrigação adequada, G1E - plantas com umaaplicação de glicina betaína e estresse hídrico, G2I - plantas com duas aplicações de glicina betaína e irrigação adequada, G2E - plantas com duas aplicações de glicina betaína e estresse hídrico. Aos seis dias após o estresse as plantas foram reidratadas. As taxas relativas de crescimento das plantas de cana-de-açúcar reduziram com o estresse hídrico. No entanto, estas reduções foram minimizadas com a aplicação foliar de glicina betaína, principalmente, com a reidratação em que a taxa relativa de crescimento reduziu apenas 19 e 18,4%, respectivamente, em G1 e G2, enquanto em G0 a redução foi de 37,2%. A glicina betaína também minimizou as reduções na produção de massa seca total sob estresse hídrico, com reduções de 5,8 e 5,9% em G1 e G2, respectivamente, enquanto em G0 foi de 15,1%. A aplicação foliar de glicina betaína melhora a taxa relativa de crescimento de plantas de cana-de-açúcar sob estresse hídrico, sobretudo após a reidratação o que diminui os efeitos negativos do estresse sobre a produção de massa seca.(AU)

The growth of sugarcane plants is greatly reduced due to the occurrence of drought stress, which ultimately affects production. Therefore, different strategies have been used to attenuate stress conditions, such as the exogenous application of glycine betaine. The objective of this work was to evaluate the growth rate and dry mass of sugarcane that received a glycine betaine application and was submitted todrought stress and rehydration. The experiment was carried out in a greenhouse with six treatments: G0I - plants without glycine betaine with adequate irrigation, G0E - plants without glycine betaine and drought stress, G1I - plants with an application of glycine betaine and adequate irrigation, G1E - plants with an application of glycine betaine and drought stress, G2I - plants with two applications of glycine betaine and adequate irrigation, G2E - plants with two applications of glycine betaine and drought stress. Six days after the stress, the plants were rehydrated. The relative growth rates of sugarcane plants decreased with the drought stress. However, these reductions were partially minimized with the foliar application of glycine betaine, mainly with the rehydration, with the relative growth rate decreasing by only 19% and 18.4%, respectively, in G1 and G2, while the reduction was 37.2% in G0. Glycine betaine also minimized reductions in total dry mass production under drought stress, with decreases of 5.8% and 5.9% in G1 and G2, respectively, while it was 15.1% in G0. The leaf application of glycine betaine improves the relative growth rate of sugarcane plants under drought stress, especially after rehydration, which decreases the negative effects of stress on dry mass production.(AU)

Effect of high hydrostatic pressure on the antioxidant capacity and peptic hydrolysis of whey proteins / Efeito da alta pressão hidrostática na hidrólise péptica das proteínas do soro de leite e na capacidade antioxidante

The effect of high hydrostatic pressure (HHP) application on whey protein concentrate was evaluated both before (pre-treatment - PT) and during (hydrolysis assisted - HA) hydrolysis processes. A factorial design 22 with 3 central points was used with pressure (100, 250, 400 MPa) and time (5, 20 and 35 minutes) as independent variables. The hydrolysis was evaluated and monitored by soluble protein, aromatic amino acid contents and RP-HPLC. ABTS and ORAC tests were used to evaluate the in vitro antioxidant capacity. The reduction of soluble protein content was approximately 20% for conventional hydrolysis and for all PT treatments up to 4 h of reaction, while HHP assisted hydrolysis at 100 MPa showed a 35% protein reduction after 35 minutes of reaction. In addition, pressurization favored peptic hydrolysis of -lactoglobulin by up to 98% and also improved the in vitro antioxidant capacity of the hydrolysates, which increased from 34.25 to 60.89 moles TE g-1 of protein in the best treatment. The results suggest that the use of HHP assisted hydrolysis favored the peptic hydrolysis, with a reduction in hydrolysis time and increased antioxidant activity.(AU)

Neste estudo, o efeito da aplicação de alta pressão hidrostática (HHP) sobre o concentrado proteico de soro de leite foi avaliado antes (pré-tratamento - PT) e durante os processos de hidrólise (assistida por hidrólise - HA). Utilizou-se o delineamento fatorial 22 com três pontos centrais, onde as variáveis independentes foram pressão (100, 250, 400 MPa) e tempo (5, 20 e 35 minutos). A hidrólise foi avaliada pelo conteúdo de proteínas solúveis e aminoácidos aromáticos, além do perfil peptídico por RP-HPLC. As análises de ABTS e ORAC foram utilizadas para avaliar a capacidade antioxidante in vitro. A redução do teor de proteína solúvel foi de aproximadamente 20% para a hidrólise convencional e para todos os pontos de PT até 4h de reação, enquanto a hidrólise assistida por HHP a 100 MPa mostrou uma redução de 35% de proteína em 35 minutos de reação. Além disso, a pressurização favoreceu a hidrólise péptica da -lactoglobulina em até 98% e também melhorou a capacidade antioxidante in vitro dos hidrolisados, que aumentaram de 34,25 para 60,89 moles de TE g-1 de proteína no melhor tratamento. Os resultados sugerem que o uso da hidrólise assistida por HHP favoreceu a hidrólise péptica, com redução no tempo de hidrólise e aumento da atividade antioxidante.(AU)

Streamlined downstream process for efficient and sustainable (Fab)2 antivenom preparation

Antivenoms are the only validated treatment against snakebite envenoming. Numerous drawbacks pertaining to their availability, safety and efficacy are becoming increasingly evident due to low sustainability of current productions. Technological innovation of procedures generating therapeutics of higher purity and better physicochemical characteristics at acceptable cost is necessary. The objective was to develop at laboratory scale a compact, feasible and economically viable platform for preparation of equine F(ab')2 antivenom against Vipera ammodytes ammodytes venom and to support it with efficiency data, to enable estimation of the process cost-effectiveness. Methods: The principle of simultaneous caprylic acid precipitation and pepsin digestion has been implemented into plasma downstream processing. Balance between incomplete IgG breakdown, F(ab')2 over-digestion and loss of the active drug's protective efficacy was achieved by adjusting pepsin to a 1:30 substrate ratio (w/w) and setting pH at 3.2. Precipitation and digestion co-performance required 2 h-long incubation at 21 °C. Final polishing was accomplished by a combination of diafiltration and flow-through chromatography. In vivo neutralization potency of the F(ab')2 product against the venom's lethal toxicity was determined. Results: Only three consecutive steps, performed under finely tuned conditions, were sufficient for preservation of the highest process recovery with the overall yield of 74%, comparing favorably to others. At the same time, regulatory requirements were met. Final product was aggregate- and pepsin-free. Its composition profile was analyzed by mass spectrometry as a quality control check. Impurities, present in minor traces, were identified mostly as IgG/IgM fragments, contributing to active drug. Specific activity of the F(ab')2 preparation with respect to the plasma was increased 3.9-fold. Conclusion: A highly streamlined mode for production of equine F(ab')2 antivenom was engineered. In addition to preservation of the highest process yield and fulfillment of the regulatory demands, performance simplicity and rapidity in the laboratory setting were demonstrated. Suitability for large-scale manufacturing appears promising.(AU)

Streamlined downstream process for efficient and sustainable (Fab)2 antivenom preparation

Antivenoms are the only validated treatment against snakebite envenoming. Numerous drawbacks pertaining to their availability, safety and efficacy are becoming increasingly evident due to low sustainability of current productions. Technological innovation of procedures generating therapeutics of higher purity and better physicochemical characteristics at acceptable cost is necessary. The objective was to develop at laboratory scale a compact, feasible and economically viable platform for preparation of equine F(ab')2 antivenom against Vipera ammodytes ammodytes venom and to support it with efficiency data, to enable estimation of the process cost-effectiveness. Methods: The principle of simultaneous caprylic acid precipitation and pepsin digestion has been implemented into plasma downstream processing. Balance between incomplete IgG breakdown, F(ab')2 over-digestion and loss of the active drug's protective efficacy was achieved by adjusting pepsin to a 1:30 substrate ratio (w/w) and setting pH at 3.2. Precipitation and digestion co-performance required 2 h-long incubation at 21 °C. Final polishing was accomplished by a combination of diafiltration and flow-through chromatography. In vivo neutralization potency of the F(ab')2 product against the venom's lethal toxicity was determined. Results: Only three consecutive steps, performed under finely tuned conditions, were sufficient for preservation of the highest process recovery with the overall yield of 74%, comparing favorably to others. At the same time, regulatory requirements were met. Final product was aggregate- and pepsin-free. Its composition profile was analyzed by mass spectrometry as a quality control check. Impurities, present in minor traces, were identified mostly as IgG/IgM fragments, contributing to active drug. Specific activity of the F(ab')2 preparation with respect to the plasma was increased 3.9-fold. Conclusion: A highly streamlined mode for production of equine F(ab')2 antivenom was engineered. In addition to preservation of the highest process yield and fulfillment of the regulatory demands, performance simplicity and rapidity in the laboratory setting were demonstrated. Suitability for large-scale manufacturing appears promising.(AU)

Use of Betaine in Post-Hatch Feed for Broiler Chicks

Post-hatch delayed placement damages the physical and physiological development of broiler chicks. This study was designed to find adequate levels of betaine inclusion in pre-hatching and pre-starter feed, in order to minimize the negative effects of post-hatch delayed placement on broiler chicks. Newly-hatched chicks were allotted in a completely randomized design, with five treatments, five replicates of ten birds each. Five dietetic levels of betaine (control, 0.070, 0.130, 0.200 and 0.260%) were used in the pre-starter ration, offered to the chicks in the transporting box and during the pre-starter phase. Performance, yolk sac retraction, plasma glucose concentration, weight and histomorphometry of the small intestine were evaluated, after 24 hours of feed access and at 7 days of age. A metabolic trial was performed from seven to ten days of age. Betaine supplementation linearly influenced the chicks ileum crypt depth after 24 hours in the transportation box. There was a quadratic effect, with an increase in feed intake up to 0.152% betaine supplementation. There was an improvement in the quality of the jejunum with betaine supplementation above 0.1%. The metabolizable coefficient of the ether extract was improved until reaching 0.163% of betaine supplementation. Betaine supplementation around 0.150% increases feed intake, the use of ether extract and interferes with the intestinal villi of chicks at seven days of age.(AU)

Use of Betaine in Post-Hatch Feed for Broiler Chicks

Post-hatch delayed placement damages the physical and physiological development of broiler chicks. This study was designed to find adequate levels of betaine inclusion in pre-hatching and pre-starter feed, in order to minimize the negative effects of post-hatch delayed placement on broiler chicks. Newly-hatched chicks were allotted in a completely randomized design, with five treatments, five replicates of ten birds each. Five dietetic levels of betaine (control, 0.070, 0.130, 0.200 and 0.260%) were used in the pre-starter ration, offered to the chicks in the transporting box and during the pre-starter phase. Performance, yolk sac retraction, plasma glucose concentration, weight and histomorphometry of the small intestine were evaluated, after 24 hours of feed access and at 7 days of age. A metabolic trial was performed from seven to ten days of age. Betaine supplementation linearly influenced the chicks ileum crypt depth after 24 hours in the transportation box. There was a quadratic effect, with an increase in feed intake up to 0.152% betaine supplementation. There was an improvement in the quality of the jejunum with betaine supplementation above 0.1%. The metabolizable coefficient of the ether extract was improved until reaching 0.163% of betaine supplementation. Betaine supplementation around 0.150% increases feed intake, the use of ether extract and interferes with the intestinal villi of chicks at seven days of age.

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)

Absence of Trichinella spiralis in swine carcasses slaughtered in the Midwestern region of Paraná state, Brazil / Ausência de Trichinella spiralis em carcaças de suínos abatidos na região Centro-Oeste do estado do Paraná, Brasil

Trichinella spiralis causes severe infections in humans, resulting in various clinical symptoms and even death, in several countries worldwide. Human trichinellosis has been reported in 55 countries, and mainly affects certain ethnic groups and tourists. It is caused by the ingestion of raw, undercooked meat or pork, or their by-products containing parasitic larvae. Infections in swine especially occur due to the parasitic contamination of food containing animal products, predation by rodents and other animals infected with T. spiralis, and due to the frequent habit of cannibalism in intensive production systems. This study aimed to evaluate the prevalence of T. spiralis in pig carcasses from a slaughterhouse located in the midwestern region of Paraná State, Brazil. Pools of 100 g of diaphragm, containing at least 1 g of tissue from each test animal, were collected, ground, and subjected to acid digestion with pepsin (1:10,000) and 25% hydrochloric acid. Sampling was performed in accordance to the methodology recommended by the European Union. All the 735,902 samples collected between 2010 and 2013, and in 2018, were negative for Trichinella spp. larvae. The pigs came from 483 farms from 90 municipalities from the three states of southern Brazil (Paraná, Santa Catarina and Rio Grande do Sul). The absence of the parasite in different animal species, reported by surveys conducted in Brazil, could confirm the country as free from trichinellosis. Although T. spiralis is not endemic to Brazil, it is necessary to subject the meat of adult pigs slaughtered here and exported to other countries to inspections and parasitological examinations. It is therefore necessary to continually inspect pig carcasses for T. spiralis in slaughterhouses worldwide.

Trichinella spiralis causa infecções graves em humanos, resultando em vários sintomas clínicos e até morte, em vários países do mundo. A triquinelose humana foi relatada em 55 países e afeta principalmente certos grupos étnicos e turistas. É causada pela ingestão de carne crua ou malpassada, ou seus subprodutos contendo larvas parasitas. As infecções em suínos ocorrem principalmente devido à contaminação parasitária de alimentos contendo produtos de origem animal, predação de roedores e outros animais infectados com T. spiralis, além do hábito frequente de canibalismo em sistemas de produção intensiva. Este trabalho objetivou avaliar a prevalência de T. spiralis em carcaças de suínos de um abatedouro localizado na região centro-oeste do Estado do Paraná. Conjuntos de 100 g de diafragma, contendo pelo menos 1 g de tecido de cada animal testado, foram coletados, triturados e submetidos a digestão ácida com pepsina (1:10.000) e ácido clorídrico a 25%. A amostragem foi realizada de acordo com a metodologia recomendada pela União Europeia. Todas as 735.902 amostras coletadas entre 2010 e 2013, e em 2018, foram negativas para larvas de Trichinella spp. Os suínos foram provenientes de 483 fazendas, de 90 municípios, dos três estados do sul do Brasil (Paraná, Santa Catarina e Rio Grande do Sul). A ausência do parasita em diferentes espécies animais, relatada por pesquisas realizadas no Brasil, poderia confirmar o país como livre de triquinelose. Embora T. spiralis não seja endêmica no Brasil, é necessário submeter a carne de suínos adultos abatidos aqui e exportados para outros países, a inspeções e exames parasitológicos. É, portanto, necessário inspecionar continuamente carcaças de suínos para T. spiralis em abatedouros de todo o mundo.

Absence of Trichinella spiralis in swine carcasses slaughtered in the Midwestern region of Paraná state, Brazil / Ausência de Trichinella spiralis em carcaças de suínos abatidos na região Centro-Oeste do estado do Paraná, Brasil

Trichinella spiralis causes severe infections in humans, resulting in various clinical symptoms and even death, in several countries worldwide. Human trichinellosis has been reported in 55 countries, and mainly affects certain ethnic groups and tourists. It is caused by the ingestion of raw, undercooked meat or pork, or their by-products containing parasitic larvae. Infections in swine especially occur due to the parasitic contamination of food containing animal products, predation by rodents and other animals infected with T. spiralis, and due to the frequent habit of cannibalism in intensive production systems. This study aimed to evaluate the prevalence of T. spiralis in pig carcasses from a slaughterhouse located in the midwestern region of Paraná State, Brazil. Pools of 100 g of diaphragm, containing at least 1 g of tissue from each test animal, were collected, ground, and subjected to acid digestion with pepsin (1:10,000) and 25% hydrochloric acid. Sampling was performed in accordance to the methodology recommended by the European Union. All the 735,902 samples collected between 2010 and 2013, and in 2018, were negative for Trichinella spp. larvae. The pigs came from 483 farms from 90 municipalities from the three states of southern Brazil (Paraná, Santa Catarina and Rio Grande do Sul). The absence of the parasite in different animal species, reported by surveys conducted in Brazil, could confirm the country as free from trichinellosis. Although T. spiralis is not endemic to Brazil, it is necessary to subject the meat of adult pigs slaughtered here and exported to other countries to inspections and parasitological examinations. It is therefore necessary to continually inspect pig carcasses for T. spiralis in slaughterhouses worldwide.(AU)

Trichinella spiralis causa infecções graves em humanos, resultando em vários sintomas clínicos e até morte, em vários países do mundo. A triquinelose humana foi relatada em 55 países e afeta principalmente certos grupos étnicos e turistas. É causada pela ingestão de carne crua ou malpassada, ou seus subprodutos contendo larvas parasitas. As infecções em suínos ocorrem principalmente devido à contaminação parasitária de alimentos contendo produtos de origem animal, predação de roedores e outros animais infectados com T. spiralis, além do hábito frequente de canibalismo em sistemas de produção intensiva. Este trabalho objetivou avaliar a prevalência de T. spiralis em carcaças de suínos de um abatedouro localizado na região centro-oeste do Estado do Paraná. Conjuntos de 100 g de diafragma, contendo pelo menos 1 g de tecido de cada animal testado, foram coletados, triturados e submetidos a digestão ácida com pepsina (1:10.000) e ácido clorídrico a 25%. A amostragem foi realizada de acordo com a metodologia recomendada pela União Europeia. Todas as 735.902 amostras coletadas entre 2010 e 2013, e em 2018, foram negativas para larvas de Trichinella spp. Os suínos foram provenientes de 483 fazendas, de 90 municípios, dos três estados do sul do Brasil (Paraná, Santa Catarina e Rio Grande do Sul). A ausência do parasita em diferentes espécies animais, relatada por pesquisas realizadas no Brasil, poderia confirmar o país como livre de triquinelose. Embora T. spiralis não seja endêmica no Brasil, é necessário submeter a carne de suínos adultos abatidos aqui e exportados para outros países, a inspeções e exames parasitológicos. É, portanto, necessário inspecionar continuamente carcaças de suínos para T. spiralis em abatedouros de todo o mundo.(AU)

Can the Fine Wheat Bran be a Betaine Source in Poultry Diets?

The effect of fine wheat bran (FWB) as a methyl donor source on performance, metabolism, body composition and blood traits of growing broilers was studied. Three hundred and twenty broilers from eight to 28 d of age, distributed in a randomized block design, with five treatments and eight replicates of eight animals each were used. The experimental diets were: NC, formulated with 72% of the Met+Cys requirement; Met, formulated with 85% of the Met+Cys equivalents by DL-methionine addition; Bet, formulated with 85% of the Met+Cys equivalents by anhydrous betaine addition; Fwb-, formulated with 72% of the Met+Cys requirement and 20% FWB; and Fwb+, formulated with 85% of the Met+Cys requirement and 20% FWB. Feed intake was reduced (p 0.05) by FWB inclusion but the feed conversion ratio was the same (p>0.05) between the positive control diets. Supplementation of DL-methionine and anhydrous betaine showed the same (p>0.05) metabolizability of nutrients. Treatments with higher DL-methionine levels (Met and Fwb+) promoted more weight of feathers (p 0.05). Animals fed with FWB showed the lowest (p 0.05) body gains. In conclusion, FWB inclusion did not promote methyl radicals supply.(AU)

Can the Fine Wheat Bran be a Betaine Source in Poultry Diets?

The effect of fine wheat bran (FWB) as a methyl donor source on performance, metabolism, body composition and blood traits of growing broilers was studied. Three hundred and twenty broilers from eight to 28 d of age, distributed in a randomized block design, with five treatments and eight replicates of eight animals each were used. The experimental diets were: NC, formulated with 72% of the Met+Cys requirement; Met, formulated with 85% of the Met+Cys equivalents by DL-methionine addition; Bet, formulated with 85% of the Met+Cys equivalents by anhydrous betaine addition; Fwb-, formulated with 72% of the Met+Cys requirement and 20% FWB; and Fwb+, formulated with 85% of the Met+Cys requirement and 20% FWB. Feed intake was reduced (p 0.05) by FWB inclusion but the feed conversion ratio was the same (p>0.05) between the positive control diets. Supplementation of DL-methionine and anhydrous betaine showed the same (p>0.05) metabolizability of nutrients. Treatments with higher DL-methionine levels (Met and Fwb+) promoted more weight of feathers (p 0.05). Animals fed with FWB showed the lowest (p 0.05) body gains. In conclusion, FWB inclusion did not promote methyl radicals supply.

Enzymatic hydrolysis (pepsin) assisted by ultrasound in the functional Properties of hydrolyzates from different collagens / Hidrólise enzimática (pepsina) assistida por ultrassom nas propriedades Funcionais de hidrolisados de diferentes colágenos

Enzymatic hydrolysis (pepsin) assisted with or without ultrasound in the functional properties of hydrolyzates from different collagens were analyzed. Degree of hydrolysis, antioxidant activity (DPPH) and antimicrobial activity (MIC) were assessed. The treatment that resulted in greater antioxidant activity for the fiber sample was with the use of 4% of enzyme and concomitant ultrasound (40.7%), leading to a degree of hydrolysis of 21.7%. For the powdered fiber sample the hydrolysis treatment with use of 4% of enzyme resulted in lower protein content (6.97mg/mL), higher degree of hydrolysis (19.9%) and greater antioxidant activity (38.6%). The hydrolyzates showed inhibitory capacity against gram-negative bacteria Salmonella choleraesuis and gram-positive bacteria Staphylococcus aureus. It can be concluded that enzymatic hydrolysis concomitant or not with the use of ultrasound increased the functionality of the fiber and powdered fiber samples, for the other samples its use as supplementary treatment was not productive, due to the worse results of antioxidant activity (DPPH) reported. However, it provided greater hydrolysis degree.(AU)

Foram avaliados os efeitos da hidrólise enzimática (pepsina) assistida com ou sem ultrassom nas propriedades funcionais de hidrolisados de diferentes colágenos. Foi analisado o grau de hidrólise, a atividade antioxidante (DPPH) e a atividade antimicrobiana (MIC). O tratamento que possibilitou maior atividade antioxidante para a amostra fibra foi com a utilização de 4% de enzima e ultrassom concomitante (40,7%), levando a um grau de hidrólise de 21,7%. Para a amostra fibra pó o tratamento de hidrólise com uso de 4% de enzima resultou em menor teor de proteína (6,97mg/mL), maior grau de hidrólise (19,9%) e maior atividade antioxidante (38,6%). Os hidrolisados mostraram capacidade inibitória contra a bactéria gram-negativa Salmonella choleraesuis e gram-positiva Staphylococcus aureus. Pode-se concluir que a hidrólise enzimática concomitante, ou não, ao uso do ultrassom apresentou aumento da funcionalidade das amostras fibra e fibra pó. Para as demais amostras, sua utilização como tratamento complementar, a hidrólise não foi interessante, devido aos piores resultados de atividade antioxidante (DPPH) encontrados. Porém, proporcionou maior grau de hidrólise.(AU)

Enzymatic hydrolysis (pepsin) assisted by ultrasound in the functional Properties of hydrolyzates from different collagens / Hidrólise enzimática (pepsina) assistida por ultrassom nas propriedades Funcionais de hidrolisados de diferentes colágenos

Enzymatic hydrolysis (pepsin) assisted with or without ultrasound in the functional properties of hydrolyzates from different collagens were analyzed. Degree of hydrolysis, antioxidant activity (DPPH) and antimicrobial activity (MIC) were assessed. The treatment that resulted in greater antioxidant activity for the fiber sample was with the use of 4% of enzyme and concomitant ultrasound (40.7%), leading to a degree of hydrolysis of 21.7%. For the powdered fiber sample the hydrolysis treatment with use of 4% of enzyme resulted in lower protein content (6.97mg/mL), higher degree of hydrolysis (19.9%) and greater antioxidant activity (38.6%). The hydrolyzates showed inhibitory capacity against gram-negative bacteria Salmonella choleraesuis and gram-positive bacteria Staphylococcus aureus. It can be concluded that enzymatic hydrolysis concomitant or not with the use of ultrasound increased the functionality of the fiber and powdered fiber samples, for the other samples its use as supplementary treatment was not productive, due to the worse results of antioxidant activity (DPPH) reported. However, it provided greater hydrolysis degree.

Foram avaliados os efeitos da hidrólise enzimática (pepsina) assistida com ou sem ultrassom nas propriedades funcionais de hidrolisados de diferentes colágenos. Foi analisado o grau de hidrólise, a atividade antioxidante (DPPH) e a atividade antimicrobiana (MIC). O tratamento que possibilitou maior atividade antioxidante para a amostra fibra foi com a utilização de 4% de enzima e ultrassom concomitante (40,7%), levando a um grau de hidrólise de 21,7%. Para a amostra fibra pó o tratamento de hidrólise com uso de 4% de enzima resultou em menor teor de proteína (6,97mg/mL), maior grau de hidrólise (19,9%) e maior atividade antioxidante (38,6%). Os hidrolisados mostraram capacidade inibitória contra a bactéria gram-negativa Salmonella choleraesuis e gram-positiva Staphylococcus aureus. Pode-se concluir que a hidrólise enzimática concomitante, ou não, ao uso do ultrassom apresentou aumento da funcionalidade das amostras fibra e fibra pó. Para as demais amostras, sua utilização como tratamento complementar, a hidrólise não foi interessante, devido aos piores resultados de atividade antioxidante (DPPH) encontrados. Porém, proporcionou maior grau de hidrólise.