Threonine-to-lysine ratio in laying hens: physiological parameters and organ weight

The objective of this study was to evaluate the effect of threonine:digestible lysine ratio in the diet on the physiological variablesand weight of organs of light laying hens. Two hundred and ten 47 week-old Dekalb White laying henswere distributed in a completely randomized design, with five levels of threonine (0.507; 0.552; 0.597; 0.642 and 0.677%) and seven replicates of six birds each. The experimental period was 10 weeks, totaling 62 days and more eight days for the animals to adapt. The physiological parameters of cloacal temperature (CT), respiratory rate (RR) and average surface temperature (AST) were recorded weekly (7:00 am, 10:00 am, 1:00 pm, 4:00 pm, and 7:00 pm); after solid and water fasting,the birds were slaughtered to assess the absolute weight of the organs. The time of day influenced (p < 0.05) the physiological parameters RR and AST, and CT showed a significant effect (p < 0.05) of increasing levels of digestible threonine. The total weight of the pancreas, proventriculus and lung showed a significant effect (p < 0.05) of the increase in the levels of digestible threonine. The respiratory rate is affected by the levels of threoninein the diet. The 0.687% level promoted hypertrophy of the pancreas, proventriculus and lung, promoting more significant activity of these organs.(AU)

Adjusted Thr: Lys Ratio Improved the Performance and Efficiency of Japanese Quail

Two hundred and forty Japanese quail aged 125 days were randomly allocated to five treatment groups based on laying (%) and quail's weight (90.71 ± 1.8% egg/day × 100 and 178.05 ± 9.38 g, respectively), each of which included six replicates of eight quails. The diets were formulated based on corn, soybean meal, and industrial amino acids. An optimal diet achieves 100% of amino acids required by the quail requirements, except for threonine. Five treatments were made: 20% less amino acid; 10% less amino acid; optimal diet; 10% more amino acid; and 20% more amino acids than those in the optimal diet. The increase in amino acid levels in a fixed Lys: amino acid ratio led to histological alterations in the liver and uterine epithelium, reduction in blood lipid peroxidation, lower hepatic HSP70 gene expression, and the performance of laying Japanese quail. The optimal diet based on the NRC with an adjusted Thr: Lys 78 ratio (Lys 1.0%) improved the performance and efficiency of Japanese quail from 125 to 230 days of age.(AU)

Effect of Sunflower Kernel Peptides Produced by Dual-Degradation on the Growth Performance, Nutrient Digestibility, and Health Status of Broilers

The purpose of this study was to investigate the effect of sunflower kernel peptides produced by enzymatic digestion, fermentation, or both on the growth performance, nutrient digestibility, and health status of broilers. Four diets contained 20% of sunflower kernel meal as its raw form (CON) or degraded by protease (ESM), Bacillus pumilus (FSM), or both (DSM). A total of 480 yellow broilers at one day old were randomly distributed to 4 groups with 6 replicates of 20 chicks each. The feeding trial lasted for 63 d. Results showed that peptide content was increased (p<0.001) from 3.97% (CON) to 32.5% (ESM), 24.2% (FSM), and 39.1% (DSM). The three sunflower peptide groups improved (p≤0.001) feed intake and body weight gain. The peptide groups increased (p≤0.015) ileal apparent digestibility of dry matter, energy, crude protein, and amino acids (methionine, lysine, tryptophan, and threonine). Furthermore, the peptide groups improved (p≤0.029) the health status by increasing serum immunoglobulins (IgA, IgG) and glutathione peroxidase. Additionally, among the peptide groups, DSM showed more pronounced effects (p<0.05) on these parameters than ESM or FSM. It is concluded that dual-degradation by enzymolysis and fermentation has a better improvement in the nutrition and application of sunflower kernel meal in broilers.(AU)

Standardized ileal amino acid digestibility of plant-based feedstuffs and phytase supplementation in broiler chicken diets / [Digestibilidade ileal estandardizada de aminoácidos de alimentos de origem vegetal e suplementação de fitase em dietas para frangos de corte]

The objective was to determine the standardized ileal amino acid digestibility (SIAAD) in soybean meal (SBM), corn + SBM, wheat + SBM and a protein free diet (PFD) associated with phytase. A total of 672 Cobb 500 (493±10g) male chicks were distributed in a completely randomized design with twelve treatments (PFD, PFD + SBM, PFD + corn + SBM and PFD + wheat + SBM, supplemented with 0, 500 and 1000 FTU), eight replicates and seven birds per cage from 14 to 23 days. Phytase supplementation at 500 FTU increased the SIAAD of methionine in SBM (P<0.05) and threonine in corn + SBM (P<0.05). There was no significant effect (P>0.05) for the SIAAD of methionine, arginine and histidine in wheat + SBM as the phytase supplementation. However, the SIAAD of lysine, threonine, isoleucine, phenylalanine, valine, cystine, alanine, aspartic acid, glutamic acid, glycine, serine and tyrosine differed (P<0.05). In general, the SIAAD for SBM, corn + SBM and wheat + SBM are 90.32, 88.65 and 83.97% (0 FTU); 91.31, 88.81 and 88.36% (500 FTU); and 91.36, 87.09 and 87.87% (1000 FTU). In conclusion, the efficacy of phytase for improve the SIAAD vary according to the feedstuff and level of supplementation.

Objetivou-se determinar a digestibilidade ileal estandardizada dos aminoácidos (SIAAD) do farelo de soja (FS), milho + FS, trigo + FS e uma dieta livre de proteína (PFD) com fitase. Um total de 672 pintos machos (493±10g), Cobb 500, foi distribuído em um delineamento inteiramente ao acaso, com 12 tratamentos (PFD, PFD + FS, PFD + milho + FS e PFD + trigo + FS, suplementados com 0, 500 e 1000 FTU), oito repetições e sete aves por gaiola, no período de 14 a 23 dias. A suplementação de fitase em 500 FTU aumentou o SIAAD da metionina do FS (P<0,05) e da treonina do milho + FS (P<0,05). Não houve efeito significativo (P>0,05) para o SIAAD de metionina, arginina e histidina no trigo + farelo de soja à medida que os níveis de fitase aumentaram. No entanto, o SIAAD de lisina, treonina, isoleucina, fenilalanina, valina, cistina, alanina, ácido aspártico, ácido glutâmico, glicina, serina e tirosina diferiu (P<0,05). Em geral, a SIAAD do FS, milho + FS e trigo + FS foi de 90,32, 88,65 e 83,97% sem a suplementação de fitase (0 FTU); 91,31, 88,81 e 88,36% (500 FTU); e 91,36, 87,09 e 87,87% (1000 FTU). Em conclusão, a eficácia da suplementação com fitase para aumentar a SIAAD varia de acordo com o alimento e o nível de suplementação.

Biochemical composition of some Echinodermata (Holothuroidea, Echinoidea) from the Red Sea, Egypt

Abstract In the present study, the biochemical composition of some Echinodermata (Holothuroidea, Echinoidea) from the Red Sea, Egypt was investigated. The results showed that the highest percentage of saturated fatty acids (SFA) was in Holothuria nobilis and the lowest percentage was recorded in Holothuria scabra. The highest percentage of monounsaturated fatty acids (MUFA) was recorded in Pearsonothuria graeffei while the lowest percentage was recorded in Holothuria atra. The highest percentage of polyunsaturated fatty acids (PUFA) was recorded in Holothuria nobilis and the lowest percentage was recorded in Holothuria atra. The highest percentage of omega-3 fatty acids was recorded in Stichopus hermanni however, the lowest percentage was recorded in Diadema setosum. The highest percentage of omega-6 fatty acids was recorded in Bohadschia vitiensis where, the lowest percentage was recorded in Holothuria atra. In addition, nonessential amino acid (NEAA) glycine were the highest in all species and serine was the lowest while Essential amino acid (EAA) arginine was the highest and cysteine was the lowest. All of the NEAA recorded highest concentrations in S. hermanni except proline. The recorded essential amino acids were histidine, isoleucine, phenylalanine, and leucine. The latter was high in Pearsonothuria graeffei. Tyrosine, valine, threonine, and arginine were high in Stichopus hermanni. The present study proved that sea cucumbers has great nutritional value.

Resumo No presente estudo, investigou-se a composição bioquímica de algumas espécies Echinodermata (Holothuroidea, Echinoidea) do Mar Vermelho no Egito. Os resultados mostraram que a maior porcentagem de ácidos graxos saturados (SFA) foi na espécie Holothuria nobilis e a menor porcentagem foi registrada em Holothuria scabra. A maior porcentagem de ácidos graxos monoinsaturados (MUFA) foi registrada na espécie Pearsonothuria graeffei, enquanto a menor porcentagem foi registrada em Holothuria atra. O maior percentual de ácidos graxos poli-insaturados (PUFA) foi registrado em Holothuria nobilis e o menor percentual foi registrado em Holothuria atra. A maior porcentagem de ácidos graxos ômega-3 foi registrada na espécie Stichopus hermanni, entretanto, a menor porcentagem foi registrada em Diadema setosum. A maior porcentagem de ácidos graxos ômega-6 foi registrada em Bohadschia vitiensis e a menor porcentagem foi registrada em Holothuria atra. Além disso, o nível de aminoácido não-essencial (NEAA) glicina foi o mais alto em todas as espécies e o de serina foi o mais baixo, enquanto o nível de aminoácido essencial (EAA) arginina foi o mais alto e a cisteína foi o mais baixo. Todos os NEAA registraram concentrações mais altas em S. hermanni, exceto prolina. Os aminoácidos essenciais registrados foram histidina, isoleucina, fenilalanina e leucina. A leucina apresentou alto nível na espécie Pearsonothuria graeffei. Tirosina, valina, treonina e arginina apresentaram níveis elevados em Stichopus hermanni. Assim, o presente estudo comprovou que o pepino-do-mar possui grande valor nutricional.

Biochemical composition of some Echinodermata (Holothuroidea, Echinoidea) from the Red Sea, Egypt / Composição bioquímica de alguns Echinodermata (Holothuroidea, Echinoidea) do Mar Vermelho, Egito

In the present study, the biochemical composition of some Echinodermata (Holothuroidea, Echinoidea) from the Red Sea, Egypt was investigated. The results showed that the highest percentage of saturated fatty acids (SFA) was in Holothuria nobilis and the lowest percentage was recorded in Holothuria scabra. The highest percentage of monounsaturated fatty acids (MUFA) was recorded in Pearsonothuria graeffei while the lowest percentage was recorded in Holothuria atra. The highest percentage of polyunsaturated fatty acids (PUFA) was recorded in Holothuria nobilis and the lowest percentage was recorded in Holothuria atra. The highest percentage of omega-3 fatty acids was recorded in Stichopus hermanni however, the lowest percentage was recorded in Diadema setosum. The highest percentage of omega-6 fatty acids was recorded in Bohadschia vitiensis where, the lowest percentage was recorded in Holothuria atra. In addition, nonessential amino acid (NEAA) glycine were the highest in all species and serine was the lowest while Essential amino acid (EAA) arginine was the highest and cysteine was the lowest. All of the NEAA recorded highest concentrations in S. hermanni except proline. The recorded essential amino acids were histidine, isoleucine, phenylalanine, and leucine. The latter was high in Pearsonothuria graeffei. Tyrosine, valine, threonine, and arginine were high in Stichopus hermanni. The present study proved that sea cucumbers has great nutritional value.

No presente estudo, investigou-se a composição bioquímica de algumas espécies Echinodermata (Holothuroidea, Echinoidea) do Mar Vermelho no Egito. Os resultados mostraram que a maior porcentagem de ácidos graxos saturados (SFA) foi na espécie Holothuria nobilis e a menor porcentagem foi registrada em Holothuria scabra. A maior porcentagem de ácidos graxos monoinsaturados (MUFA) foi registrada na espécie Pearsonothuria graeffei, enquanto a menor porcentagem foi registrada em Holothuria atra. O maior percentual de ácidos graxos poliinsaturados (PUFA) foi registrado em Holothuria nobilis e o menor percentual foi registrado em Holothuria atra. A maior porcentagem de ácidos graxos ômega-3 foi registrada na espécie Stichopus hermanni, entretanto, a menor porcentagem foi registrada em Diadema setosum. A maior porcentagem de ácidos graxos ômega-6 foi registrada em Bohadschia vitiensis e a menor porcentagem foi registrada em Holothuria atra. Além disso, o nível de aminoácido não-essencial (NEAA) glicina foi o mais alto em todas as espécies e o de serina foi o mais baixo, enquanto o nível de aminoácido essencial (EAA) arginina foi o mais alto e a cisteína foi o mais baixo. Todos os NEAA registraram concentrações mais altas em S. hermanni, exceto prolina. Os aminoácidos essenciais registrados foram histidina, isoleucina, fenilalanina e leucina. A leucina apresentou alto nível na espécie Pearsonothuria graeffei. Tirosina, valina, treonina e arginina apresentaram níveis elevados em Stichopus hermanni. Assim, o presente estudo comprovou que o pepino-do-mar possui grande valor nutricional.

Biochemical composition of some Echinodermata (Holothuroidea, Echinoidea) from the Red Sea, Egypt / Composição bioquímica de alguns Echinodermata (Holothuroidea, Echinoidea) do Mar Vermelho, Egito

In the present study, the biochemical composition of some Echinodermata (Holothuroidea, Echinoidea) from the Red Sea, Egypt was investigated. The results showed that the highest percentage of saturated fatty acids (SFA) was in Holothuria nobilis and the lowest percentage was recorded in Holothuria scabra. The highest percentage of monounsaturated fatty acids (MUFA) was recorded in Pearsonothuria graeffei while the lowest percentage was recorded in Holothuria atra. The highest percentage of polyunsaturated fatty acids (PUFA) was recorded in Holothuria nobilis and the lowest percentage was recorded in Holothuria atra. The highest percentage of omega-3 fatty acids was recorded in Stichopus hermanni however, the lowest percentage was recorded in Diadema setosum. The highest percentage of omega-6 fatty acids was recorded in Bohadschia vitiensis where, the lowest percentage was recorded in Holothuria atra. In addition, nonessential amino acid (NEAA) glycine were the highest in all species and serine was the lowest while Essential amino acid (EAA) arginine was the highest and cysteine was the lowest. All of the NEAA recorded highest concentrations in S. hermanni except proline. The recorded essential amino acids were histidine, isoleucine, phenylalanine, and leucine. The latter was high in Pearsonothuria graeffei. Tyrosine, valine, threonine, and arginine were high in Stichopus hermanni. The present study proved that sea cucumbers has great nutritional value.

No presente estudo, investigou-se a composição bioquímica de algumas espécies Echinodermata (Holothuroidea, Echinoidea) do Mar Vermelho no Egito. Os resultados mostraram que a maior porcentagem de ácidos graxos saturados (SFA) foi na espécie Holothuria nobilis e a menor porcentagem foi registrada em Holothuria scabra. A maior porcentagem de ácidos graxos monoinsaturados (MUFA) foi registrada na espécie Pearsonothuria graeffei, enquanto a menor porcentagem foi registrada em Holothuria atra. O maior percentual de ácidos graxos poli insaturados (PUFA) foi registrado em Holothuria nobilis e o menor percentual foi registrado em Holothuria atra. A maior porcentagem de ácidos graxos ômega-3 foi registrada na espécie Stichopus hermanni, entretanto, a menor porcentagem foi registrada em Diadema setosum. A maior porcentagem de ácidos graxos ômega-6 foi registrada em Bohadschia vitiensis e a menor porcentagem foi registrada em Holothuria atra. Além disso, o nível de aminoácido não-essencial (NEAA) glicina foi o mais alto em todas as espécies e o de serina foi o mais baixo, enquanto o nível de aminoácido essencial (EAA) arginina foi o mais alto e a cisteína foi o mais baixo. Todos os NEAA registraram concentrações mais altas em S. hermanni, exceto prolina. Os aminoácidos essenciais registrados foram histidina, isoleucina, fenilalanina e leucina. A leucina apresentou alto nível na espécie Pearsonothuria graeffei. Tirosina, valina, treonina e arginina apresentaram níveis elevados em Stichopus hermanni. Assim, o presente estudo comprovou que o pepino-do-mar possui grande valor nutricional.

Biochemical composition of some Echinodermata (Holothuroidea, Echinoidea) from the Red Sea, Egypt / Composição bioquímica de alguns Echinodermata (Holothuroidea, Echinoidea) do Mar Vermelho, Egito

In the present study, the biochemical composition of some Echinodermata (Holothuroidea, Echinoidea) from the Red Sea, Egypt was investigated. The results showed that the highest percentage of saturated fatty acids (SFA) was in Holothuria nobilis and the lowest percentage was recorded in Holothuria scabra. The highest percentage of monounsaturated fatty acids (MUFA) was recorded in Pearsonothuria graeffei while the lowest percentage was recorded in Holothuria atra. The highest percentage of polyunsaturated fatty acids (PUFA) was recorded in Holothuria nobilis and the lowest percentage was recorded in Holothuria atra. The highest percentage of omega-3 fatty acids was recorded in Stichopus hermanni however, the lowest percentage was recorded in Diadema setosum. The highest percentage of omega-6 fatty acids was recorded in Bohadschia vitiensis where, the lowest percentage was recorded in Holothuria atra. In addition, nonessential amino acid (NEAA) glycine were the highest in all species and serine was the lowest while Essential amino acid (EAA) arginine was the highest and cysteine was the lowest. All of the NEAA recorded highest concentrations in S. hermanni except proline. The recorded essential amino acids were histidine, isoleucine, phenylalanine, and leucine. The latter was high in Pearsonothuria graeffei. Tyrosine, valine, threonine, and arginine were high in Stichopus hermanni. The present study proved that sea cucumbers has great nutritional value.(AU)

No presente estudo, investigou-se a composição bioquímica de algumas espécies Echinodermata (Holothuroidea, Echinoidea) do Mar Vermelho no Egito. Os resultados mostraram que a maior porcentagem de ácidos graxos saturados (SFA) foi na espécie Holothuria nobilis e a menor porcentagem foi registrada em Holothuria scabra. A maior porcentagem de ácidos graxos monoinsaturados (MUFA) foi registrada na espécie Pearsonothuria graeffei, enquanto a menor porcentagem foi registrada em Holothuria atra. O maior percentual de ácidos graxos poli insaturados (PUFA) foi registrado em Holothuria nobilis e o menor percentual foi registrado em Holothuria atra. A maior porcentagem de ácidos graxos ômega-3 foi registrada na espécie Stichopus hermanni, entretanto, a menor porcentagem foi registrada em Diadema setosum. A maior porcentagem de ácidos graxos ômega-6 foi registrada em Bohadschia vitiensis e a menor porcentagem foi registrada em Holothuria atra. Além disso, o nível de aminoácido não-essencial (NEAA) glicina foi o mais alto em todas as espécies e o de serina foi o mais baixo, enquanto o nível de aminoácido essencial (EAA) arginina foi o mais alto e a cisteína foi o mais baixo. Todos os NEAA registraram concentrações mais altas em S. hermanni, exceto prolina. Os aminoácidos essenciais registrados foram histidina, isoleucina, fenilalanina e leucina. A leucina apresentou alto nível na espécie Pearsonothuria graeffei. Tirosina, valina, treonina e arginina apresentaram níveis elevados em Stichopus hermanni. Assim, o presente estudo comprovou que o pepino-do-mar possui grande valor nutricional.(AU)

Multivariate modeling strategies to predict nutritional requirements of essential amino acids in semiheavy second-cycle hens

An experiment with 23 diets was performed to evaluate the effect of digestible lysine (Lys), digestible methionine + cysteine (Met+Cys), and digestible threonine (Thr) on egg production of H&N Brown second-cycle laying hens (SCLH) for 20 weeks (92-111 weeks of age) in cages under environmental conditions. Body weight (BW), feed intake (FI), feed conversion ratio (FCR), egg weight (EW), number of hen-housed eggs, and livability were also evaluated during the experiment. Diets were formulated from a central composite design that combined five levels of Lys, Met+Cys, and Thr ranging from 727 to 1159, 662 to 1055, and 552 to 882 mg/kg, respectively. Egg production (EP) data were evaluated through three different modeling strategies: egg production models, multivariate polynomial models, and artificial neural networks (ANN). A cascade-forward neural network with logsigmoid transfer function was selected as the best model according to goodness-offit statistics in both identification and validation data. One of the best scenarios for EP of H&N Brown SCLH under specific outdoor conditions was established at Lys, Met+Cys, and Thr levels of 1138, 1031, and 717 mg/hen·day, respectively. The ANN model may be an appropriate tool to study and predict EP of H&N Brown SCLH based on the combination of three different levels of essential digestible amino acids. The strategies included in this work may contribute to improving poultry performance based on modeling techniques to study other production parameters in terms of different nutritional requirements and productive conditions.

Use of protein kinase C and phospholipase A2 inhibitors in bovine endometrial cells treated with estradiol and calcium ionophore / Uso de inibidores de proteína quinase C e fosfolipase A2 em células endometriais bovinas tratadas com estradiol e ionóforo de cálcio

The release of endometrial prostaglandin-F2α (PGF2α) in bovine females can be induced in vivo by estradiol (E2). However, its role in this mechanism has not been clarified. We hypothesized that E2 stimulates the activity and abundance of protein kinase C (PKC) and phospholipase A2 (PLA2). Our objective in this study was to analyze the effects of PKC and PLA2 inhibitors on PGF2α synthesis induced by E2 and calcium ionophore (CI) in bovine endometrial cells (BEND cells; Experiment 1). Additionally, we evaluated the abundance of PKC and PLA2 in endometrial explants of cows treated or not with E2 17 days after estrus (D17, D0 = estrus; Experiment 2). In Experiment 1, BEND cells were submitted to a PKC inhibitor (10 µM of C25H24N4O2; bisindolylmaleimide I, or BIS I), a PLA2 inhibitor (20 µM of arachydoniltrifluoromethane or AACOCF3), or none. The BEND cells were subsequently treated with E2 and CI, and PGF2α concentrations were measured in the culture medium through radioimmunoassay. For DIF-12 (PGF2α concentration 12 h after treatment subtracted from PGF2α concentration at hour 0), no PKC inhibitor effect was observed (P= 0.2709). However, DIF-12 was lower (P < 0.05) for groups treated with the PLA2 inhibitor and PLA2 inhibitor + CI + E2 groups than the control and CI + E2 groups. Thus, AACOCF3 was an efficient PLA2 inhibitor in the BEND cells culture system, and E2 did not stimulate the synthesis of PKC and PLA2. In Experiment 2, cyclic Nellore heifers received none (n = 5) or 3 mg (n = 6) of 17ß-E2 on D17 and were slaughtered 2 h after administration. The abundance of PKC and PLA2 in the endometrial tissue was evaluated using Western blotting analysis. No E2 effect was observed on PKC (P = 0.08) and PLA2 (P = 0.56). We concluded that E2 did not stimulate the activity and abundance of PKC and PLA2.(AU)

A liberação endometrial de prostaglandina-F2α (PGF2α) em fêmeas bovinas pode ser induzida in vivo pelo estradiol (E2). Entretanto o seu mecanismo de ação ainda não foi bem esclarecido. Nossa hipótese é que o E2 estimula a atividade e a abundância da proteína quinase C (PKC) e da fosfolipase A2 (PLA2). Nosso objetivo com este estudo foi analizar os efeitos de inibidores de PKC e PLA2 na síntese de PGF2α induzida por E2 e ionóforo de cálcio (CI) em células endometriais bovinas (células BEND; Experimento 1). Adicionalmente, nós avaliamos a abundância de PKC e PLA2 em explantes endometriais de vacas tratadas com ou sem E2 17 dias após o estro (D17, D0 = estro; Experimento 2). No Experimento 1, células BEND foram submetidas ao inibidor de PKC (10 µM de C25H24N4O2; bisindolylmaleimide I, ou BIS I), e ao inibidor de PLA2 (20 µM de arachydoniltrifluoromethane ou AACOCF3) ou a nenhum inibidor. As células BEND foram subsequentemente tratadas com E2 e CI e concentrações de PGF2α foram mensuradas no meio de cultura por radioimunoenssaio. Para DIF-12 (concentração de PGF2α 12 horas depois do tratamento, subtraída da concentração de PGF2α na hora 0), não foi observado efeito do inibidor de PKC (P = 0.2709). Entretanto DIF-12 foi menor (P < 0.05) nos grupos tratados com inibidor de PLA2 e inibidor de PLA2 + CI + E2 quando comparados com o grupo controle e o grupo CI + E2. O AACOCF3 foi um eficiente inibidor de PLA2 em sistema de cultura de células BEND e o E2 não estimulou a síntese de PKC e PLA2. No Experimento 2, novilhas Nelore cíclicas receberam 3 mg de 17ß-E2 (n = 6) ou nenhum tratamento (n = 5) no D17 e foram abatidas duas horas depois da administração dos tratamentos. A quantidade de PKC and PLA2 no tecido endometrial foi avaliada pela técnica de Western Blotting. Não foi observado efeito do E2 sobre a PKC (P= 0.08) e nem sobre a PLA2 (P= 0.56). Conclui-se que o E2 não estimulou a atividade e abundância de PKC e PLA2.(AU)

Use of protein kinase C and phospholipase A2 inhibitors in bovine endometrial cells treated with estradiol and calcium ionophore / Uso de inibidores de proteína quinase C e fosfolipase A2 em células endometriais bovinas tratadas com estradiol e ionóforo de cálcio

The release of endometrial prostaglandin-F2α (PGF2α) in bovine females can be induced in vivo by estradiol (E2). However, its role in this mechanism has not been clarified. We hypothesized that E2 stimulates the activity and abundance of protein kinase C (PKC) and phospholipase A2 (PLA2). Our objective in this study was to analyze the effects of PKC and PLA2 inhibitors on PGF2α synthesis induced by E2 and calcium ionophore (CI) in bovine endometrial cells (BEND cells; Experiment 1). Additionally, we evaluated the abundance of PKC and PLA2 in endometrial explants of cows treated or not with E2 17 days after estrus (D17, D0 = estrus; Experiment 2). In Experiment 1, BEND cells were submitted to a PKC inhibitor (10 µM of C25H24N4O2; bisindolylmaleimide I, or BIS I), a PLA2 inhibitor (20 µM of arachydoniltrifluoromethane or AACOCF3), or none. The BEND cells were subsequently treated with E2 and CI, and PGF2α concentrations were measured in the culture medium through radioimmunoassay. For DIF-12 (PGF2α concentration 12 h after treatment subtracted from PGF2α concentration at hour 0), no PKC inhibitor effect was observed (P= 0.2709). However, DIF-12 was lower (P < 0.05) for groups treated with the PLA2 inhibitor and PLA2 inhibitor + CI + E2 groups than the control and CI + E2 groups. Thus, AACOCF3 was an efficient PLA2 inhibitor in the BEND cells culture system, and E2 did not stimulate the synthesis of PKC and PLA2. In Experiment 2, cyclic Nellore heifers received none (n = 5) or 3 mg (n = 6) of 17ß-E2 on D17 and were slaughtered 2 h after administration. The abundance of PKC and PLA2 in the endometrial tissue was evaluated using Western blotting analysis. No E2 effect was observed on PKC (P = 0.08) and PLA2 (P = 0.56). We concluded that E2 did not stimulate the activity and abundance of PKC and PLA2.(AU)

A liberação endometrial de prostaglandina-F2α (PGF2α) em fêmeas bovinas pode ser induzida in vivo pelo estradiol (E2). Entretanto o seu mecanismo de ação ainda não foi bem esclarecido. Nossa hipótese é que o E2 estimula a atividade e a abundância da proteína quinase C (PKC) e da fosfolipase A2 (PLA2). Nosso objetivo com este estudo foi analizar os efeitos de inibidores de PKC e PLA2 na síntese de PGF2α induzida por E2 e ionóforo de cálcio (CI) em células endometriais bovinas (células BEND; Experimento 1). Adicionalmente, nós avaliamos a abundância de PKC e PLA2 em explantes endometriais de vacas tratadas com ou sem E2 17 dias após o estro (D17, D0 = estro; Experimento 2). No Experimento 1, células BEND foram submetidas ao inibidor de PKC (10 µM de C25H24N4O2; bisindolylmaleimide I, ou BIS I), e ao inibidor de PLA2 (20 µM de arachydoniltrifluoromethane ou AACOCF3) ou a nenhum inibidor. As células BEND foram subsequentemente tratadas com E2 e CI e concentrações de PGF2α foram mensuradas no meio de cultura por radioimunoenssaio. Para DIF-12 (concentração de PGF2α 12 horas depois do tratamento, subtraída da concentração de PGF2α na hora 0), não foi observado efeito do inibidor de PKC (P = 0.2709). Entretanto DIF-12 foi menor (P < 0.05) nos grupos tratados com inibidor de PLA2 e inibidor de PLA2 + CI + E2 quando comparados com o grupo controle e o grupo CI + E2. O AACOCF3 foi um eficiente inibidor de PLA2 em sistema de cultura de células BEND e o E2 não estimulou a síntese de PKC e PLA2. No Experimento 2, novilhas Nelore cíclicas receberam 3 mg de 17ß-E2 (n = 6) ou nenhum tratamento (n = 5) no D17 e foram abatidas duas horas depois da administração dos tratamentos. A quantidade de PKC and PLA2 no tecido endometrial foi avaliada pela técnica de Western Blotting. Não foi observado efeito do E2 sobre a PKC (P= 0.08) e nem sobre a PLA2 (P= 0.56). Conclui-se que o E2 não estimulou a atividade e abundância de PKC e PLA2.(AU)

Nutritive evaluation, metabolisable energy and digestible amino acid contents of different indigenous feedstuff for Nile tilapia (Oreochromis niloticus) / Avaliação nutricional, energia metabolizável e conteúdo de aminoácidos digestíveis de diferentes alimentos indígenas para tilápia do Nilo (Oreochromis niloticus)

Three trials were executed to examine the nutritive profile, metabolisable energy and digestible amino acid (AA) contents of four indigenous feed ingredients including wheat (W), wheat middling (WM), canola meal (CM) and rapeseed meal (RSM) in Nile tilapia. Three samples of each test ingredient were collected from three different locations of Multan (MUL) and Sukkar (SKR), of Pakistan. The collected three samples were pooled thereafter to make a homogenous/ representative sample of each test ingredient from a particular study site. Nutrients composition, AA and energy digestibility of these indigenous ingredients were evaluated by using laboratory analyses and fish studies. Proximate analysis indicated variations in some of the nutrients due to location (p < 0.05). Differences were also observed in some AA including arginine, lysine, serine, cysteine, glutamic and aspartic acids, histidine, valine and glycine contents of these ingredients (p < 0.05). Digestibility of leucine, glycine and glutamic acid was higher (p < 0.05) in RSM from MUL. Among W samples from MUL, AA digestibility for lysine, threonine, and aspartic acid was higher (p < 0.05). Crude protein, arginine, alanine, serine, and aspartic acid had higher digestibility (p < 0.05), whereas digestibility was lower (p < 0.05) for threonine, valine and tyrosine in RSM from MUL. Metaboliseable energy contents did not differ among W, WM, CM and RSM regarding their origin (p > 0.05). The results indicated that nutritional profiles and their digestibility indices vary with the location for Nile tilapia.(AU)

Três experimentos foram executados para examinar o perfil nutritivo, a energia metabolizável e o conteúdo de aminoácidos digestíveis (AA) de quatro ingredientes alimentícios, incluindo trigo (W), farelo de trigo (WM), farelo de canola (CM) e farelo de colza (RSM) em tilápia do Nilo. Três amostras de cada ingrediente do teste foram coletadas de dois locais diferentes (Multan (MUL) e Sukkar (SKR), do Paquistão) e assim agrupadas. A composição nutricional, AA e digestibilidade energética desses ingredientes indígenas foram avaliadas por meio de análises laboratoriais e estudos de peixes. A análise imediata indicou variações (p < 0,05) em alguns dos nutrientes devido à localização. Variações (p < 0,05) também foram observadas em alguns teores de AA desses ingredientes. A digestibilidade da leucina, glicina e ácido glutâmico foi maior (p < 0,05) em RSM de MUL. Entre as amostras de W da MUL, a digestibilidade de AA para Lys, Thr e Asp foi maior (p < 0,05). Proteína Crud, arginina, alanina, serina e ácido aspártico apresentaram maior digestibilidade (p < 0,05), e menor (p < 0,05) para treonina, valina e tirosina em MRS. Nenhuma diferença (p> 0,05) relacionada a energia metabolizável foi observada entre esses ingredientes em relação à sua origem. Os resultados indicaram que os perfis nutricionais e sua digestibilidade variam com a localização.(AU)

The role of Rho-associated kinase inhibitor, Y-27632 on primary culture of ovine spermatogonial stem cells

Abstract The access to sufficient numbers of spermatogonial stem cells (SSCs) is a prerequisite for the study of their regulation and further biomanipulation. Rho kinase (ROCK) belongs to a family of serine/threonine kinases and involves in a wide range of fundamental cellular functions. The aim of the present study was to study the effect of ROCK inhibitor, Y-27632 (0.1-40 µM), during the primary culture of ovine SSCs. SSCs were collected from 3-5-month-old's lamb testes. The viability of SSCs, the apoptosis assay of SSCs, the intracellular reactive oxygen species (ROS) analysis, and the SSCs markers and apoptosis-related gene expressions were detected by MTT reduction assay, Annexin V-FITC/ Propidium Iodide (PI) dual staining, flow cytometry and real-time-PCR studies, respectively. Morphological analyses indicated that the 5-10 µM Y-27632 had an optimal effect on the number of presumptive SSCs colonies and the area covered by them after a 10 days culture. The cell viability, apoptosis and necrosis of SSCs after 10 days' culture were not affected in comparison with the control group, and the 20 µM of Y-27632 resulted in significantly decreased cell viability (P<0.05) and an increased necrosis of cells. On day 10 after culture, the expression of P53 was decreased with an increase from 0 to 10 µM in the Y-27632 dose. In the 20 µM Y-27632 group, the expressions of P53 and Bax were higher and the Bcl-2 was lower than other groups and these values were significantly different from 5 and 10 µM Y-27632 groups (P<0.05). The level of intracellular ROS was decreased with an increase in the Y-27632 dose from 5 to 20 µM in comparison with the control group. In conclusion, the present study demonstrated that Y-27632 at a concentration of 5-10 µM provided optimal culture conditions for the primary culture of ovine SSCs.

The neuroprotection of cerebrolysin after spontaneous intracerebral hemorrhage through regulates necroptosis via Akt/ GSK3B signaling pathway

ABSTRACT Purpose: Spontaneous intracerebral hemorrhage (ICH) is a major cause of death and disability with a huge economic burden worldwide. Cerebrolysin (CBL) has been previously used as a nootropic drug. Necroptosis is a programmed cell death mechanism that plays a vital role in neuronal cell death after ICH. However, the precise role of necroptosis in CBL neuroprotection following ICH has not been confirmed. Methods: In the present study, we aimed to investigate the neuroprotective effects and potential molecular mechanisms of CBL in ICH-induced early brain injury (EBI) by regulating neural necroptosis in the C57BL/6 mice model. Mortality, neurological score, brain water content, and neuronal death were evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, Evans blue extravasation, Western blotting, and quantitative real-time polymerase chain reaction (PCR). Results: The results show that CBL treatment markedly increased the survival rate, neurological score, and neuron survival, and downregulated the protein expression of RIP1 and RIP3, which indicated that CBL-mediated inhibition of necroptosis, and ameliorated neuronal death after ICH. The neuroprotective capacity of CBL is partly dependent on the Akt/GSK3β signaling pathway. Conclusions: CBL improves neurological outcomes in mice and reduces neuronal death by protecting against neural necroptosis.

The role of Rho-associated kinase inhibitor, Y-27632 on primary culture of ovine spermatogonial stem cells

The access to sufficient numbers of spermatogonial stem cells (SSCs) is a prerequisite for the study of their regulation and further biomanipulation. Rho kinase (ROCK) belongs to a family of serine/threonine kinases and involves in a wide range of fundamental cellular functions. The aim of the present study was to study the effect of ROCK inhibitor, Y-27632 (0.1-40 µM), during the primary culture of ovine SSCs. SSCs were collected from 3-5-month-olds lamb testes. The viability of SSCs, the apoptosis assay of SSCs, the intracellular reactive oxygen species (ROS) analysis, and the SSCs markers and apoptosis-related gene expressions were detected by MTT reduction assay, Annexin V–FITC/ Propidium Iodide (PI) dual staining, flow cytometry and real-time-PCR studies, respectively. Morphological analyses indicated that the 5-10 µM Y-27632 had an optimal effect on the number of presumptive SSCs colonies and the area covered by them after a 10 days culture. The cell viability, apoptosis and necrosis of SSCs after 10 days culture were not affected in comparison with the control group, and the 20 µM of Y-27632 resulted in significantly decreased cell viability (P<0.05) and an increased necrosis of cells. On day 10 after culture, the expression of P53 was decreased with an increase from 0 to 10 µM in the Y-27632 dose. In the 20 µM Y-27632 group, the expressions of P53 and Bax were higher and the Bcl-2 was lower than other groups and these values were significantly different from 5 and 10 µM Y-27632 groups (P<0.05). The level of intracellular ROS was decreased with an increase in the Y-27632 dose from 5 to 20 µM in comparison with the control group. In conclusion, the present study demonstrated that Y-27632 at a concentration of 5-10 µM provided optimal culture conditions for the primary culture of ovine SSCs.(AU)

Evaluation of Muscle Chemical and Amino Acids Composition in Broiler Chicks Fed Sorghum or Sorghum-Pea Diets

The study evaluated the chemical and amino acids (AA) composition of breast and thigh muscle in broilers fed sorghum and sorghum-pea diets, as partial substitute of corn and soybean meal (SBM). A total of 540 3-wk-old broilers (Cobb 500) randomly assigned to three groups were fed with corn-SBM control diet (C), corn-sorghum-SBM diet (S) and corn-sorghum-peas-SBM diet (SP) for finisher phase. At slaughter, muscle samples were collected for chemical analyses. The results showed that dietary sorghum or sorghum-pea inclusion did not affect (p>0.05) the chemical composition (dry matter, protein, fat and ash) of broilers muscle tissue. The total AA (TAA), essential AA (EAA) or flavour-related AA (FAA) concentrations from breast or thigh muscle did not differ (p>0.05) between treatments. A significant effect (p<0.05) was found for some individual EAA. The valine and phenylalanine concentrations were higher, and cysteine and methionine levels were lower in both muscles than the C group. The higher deposition (p<0.05) was found for most AA, except glycine and arginine, in the breast vs thigh, as an effect of muscle tissue. As results, the TAA, EAA, NEAA, FAA and EAA/NEAA ratio increased in the breast vs thigh muscle. Interactions between diets and muscle tissue were noticed for serine, threonine, methionine, isoleucine, phenylalanine, lysine, arginine, TAA and EAA. In conclusion, sorghum or sorghum-peas can partially substitute the corn and SBM in broiler diets, with no adverse effects on chemical composition and beneficial nutrients, such as EAA and FAA that are important for the nutritional quality of meat.(AU)

Evaluation of Muscle Chemical and Amino Acids Composition in Broiler Chicks Fed Sorghum or Sorghum-Pea Diets

The study evaluated the chemical and amino acids (AA) composition of breast and thigh muscle in broilers fed sorghum and sorghum-pea diets, as partial substitute of corn and soybean meal (SBM). A total of 540 3-wk-old broilers (Cobb 500) randomly assigned to three groups were fed with corn-SBM control diet (C), corn-sorghum-SBM diet (S) and corn-sorghum-peas-SBM diet (SP) for finisher phase. At slaughter, muscle samples were collected for chemical analyses. The results showed that dietary sorghum or sorghum-pea inclusion did not affect (p>0.05) the chemical composition (dry matter, protein, fat and ash) of broilers muscle tissue. The total AA (TAA), essential AA (EAA) or flavour-related AA (FAA) concentrations from breast or thigh muscle did not differ (p>0.05) between treatments. A significant effect (p<0.05) was found for some individual EAA. The valine and phenylalanine concentrations were higher, and cysteine and methionine levels were lower in both muscles than the C group. The higher deposition (p<0.05) was found for most AA, except glycine and arginine, in the breast vs thigh, as an effect of muscle tissue. As results, the TAA, EAA, NEAA, FAA and EAA/NEAA ratio increased in the breast vs thigh muscle. Interactions between diets and muscle tissue were noticed for serine, threonine, methionine, isoleucine, phenylalanine, lysine, arginine, TAA and EAA. In conclusion, sorghum or sorghum-peas can partially substitute the corn and SBM in broiler diets, with no adverse effects on chemical composition and beneficial nutrients, such as EAA and FAA that are important for the nutritional quality of meat.

Influence of a blend of dietary essential amino acids on growth, enzymatic activity, and intestinal histology of blue discus / Influência de um “blend” de aminoácidos essenciais sobre o crescimento, atividade enzimática e histologia intestinal de acará disco

Symphysodon aequifasciatus is a fish with a disk-shaped body and bright colors, important characteristics of ornamental fish. We evaluated amino acid supplementation strategies to reduce crude protein in the diet for evaluation of performance, the content of digestive enzymes, liver metabolism, and intestinal histopathology. A total of 180 fish were randomly distributed in 12 separate 50 L glass aquariums, consisting of a completely randomized design with four treatments (DC - Control diet with 34.4% crude protein; DL - Control diet plus 1% of lysine; DEAA - Control diet plus 1% free essential amino acids (threonine, phenylalanine, leucine, valine, arginine, and tryptophan); and DHP - Diet with a high level of crude protein 48.4%), three repetitions, lasting 60 days. The use of DL and DEAA diets resulted in higher intestinal villus height and higher zootechnical performance. The use of DL diet increased alkaline phosphatase and digestive amylase activity. The use of DHP diets promotes severe liver changes due to increased activity of Alanine aminotraserase. Therefore, it was possible to observe that the use of amino acids can supply the nutritional need of blue discus. Supplementation of diets with AAs allows the reduction of dietary protein, which is a strategy for feeding management.

Symphysodon aequifasciatus é um peixe com corpo em forma de disco e cores brilhantes, características importantes dos peixes ornamentais. Avaliamos estratégias de suplementação de aminoácidos (AAs) para reduzir a proteína bruta na dieta com relação ao desempenho, atividade das enzimas digestivas, metabolismo hepático e histopatologia intestinal. Um total de 180 peixes foram distribuídos aleatoriamente em 12 aquários de vidro de 50 L, consistindo em um delineamento inteiramente casualizado, com quatro tratamentos (DC – Dieta controle com 34,4% de proteina bruta; DL - Dieta controle acrescida de 1% de lisina; DEAA – Dieta controle acrescida de 1% aminoácidos essenciais livres (treonina, fenilalanina, leucina, valina, arginina e triptofano) e DHP – Dieta com alto nível de proteína bruta 48,4%), e três repetições, com duração de 60 dias. Os resultados demostraram que a utilização das dietas DL e DEAA refletiram em maior altura de vilosidade intestinal e maior desempenho zootécnico. A utilização de dieta DL promoveu aumento da atividade da fosfatase alcalina e amilase digestiva. A utilização de dietas DHP resultou em alterações hepáticas graves, devido a maior atividade da Alanina aminotransferase. Portanto, foi possível observar que a utilização de aminoácidos pode suprir a necessidade nutricional de acará disco. A suplementação de dietas com AAs permite a redução da proteína dietética, o que é uma estratégia para o manejo alimentar.

Presença da mutação do gene BRCA1 em cadelas com tumores mamários malignos / Presence of BRCA1 gene mutation in bitches with malignant mammary tumors

Mammary tumors (MTs) in bitches are similar to breast cancers in women. Thus, they can be used as a model for human breast cancer and findings can be extrapolated for use in human medicine. BRCA1 is a tumor suppressor gene. When the gene has a mutation, it cannot repair damaged DNA, which causes genetic instability and tumorigenesis. Therefore, we aimed to study the frequency of single nucleotide polymorphisms (SNPs) in the BRCA1 gene that are associated with distinct histological types of malignant MT in bitches. The study population consisted of 91 bitches, including a control group of 6 animals with healthy mammary glands and 85 animals with MTs. All animals underwent a presurgery evaluation consisting of a questionnaire administered to the person responsible for the animal, a physical examination, collection of peripheral blood for hematological and serum biochemistry evaluations, an electrocardiogram, and a preanesthesia evaluation. In addition, distant metastasis was studied via chest radiography and abdominal ultrasound. After evaluations were complete, the animals that could undergo surgery were administered general anesthesia and underwent a mastectomy or mammary gland sample collection. Histopathological examination and molecular analysis were performed to identify mutations in the BRCA1 gene. Histopathological examinations found 10 different types of malignant tumors in 36 sick animals. Tumor samples plus samples from the 6 control animals were subjected to DNA extraction, polymerase chain reaction (PCR) analysis, and genetic sequencing. The tumor with the highest incidence (33.33%) was a complex carcinoma, followed by carcinoma in mixed tumor (13.88), tubular carcinoma (13.88) and carcinosarcoma (13.88). Molecular analysis revealed 3 different SNP points in 5 samples (4006G>A, 3619A>G, and 3761C>T). The allelic variant 4006G>A (1/36) resulted in the alteration of the amino acid valine by isoleucine (V1336 I). The mutation 3619A>G (2/36) inserted the amino acid alanine instead of threonine (T1207 A). The mutation 3761C>T (2/36) led to the alteration of the amino acid serine by phenylalanine (S1254 F), a mutation for which there are no published reports. The histological types that showed BRCA1 mutations were complex carcinoma (1/5), carcinoma in mixed tumor (1/5), papillary carcinoma (1/5) and tubular carcinoma (2/5). Software analysis identified the new SNP (nucleotide 3761) in BRCA1 and 2 point mutations in nucleotides 4006 and 3619 and responsible for genetic instability. The development of breast cancer is caused by many endogenous and exogenous factors. The results of our study show that these factors have a greater presence in female, mixed breed, uncastrated, and older dogs, confirming the data in the veterinary literature. In the present study, we found different histological types of malignant breast tumors with mutations in the BRCA1 gene, as other authors have reported. However, we also found the mutation 3761C>T, which, to the best of our knowledge, has not been reported in the literature. This shows the need for studies in veterinary medicine that assess mutations in the BRCA1 gene and the most common histological types. In conclusion, SNPs in the BRCA1 gene cause genetic instability, resulting in additional mutations that lead to the development of breast tumors. They are point mutations that affect transcription, resulting in truncated proteins. These proteins may have a loss of function, leading to carcinogenesis.(AU)

Influence of a blend of dietary essential amino acids on growth, enzymatic activity, and intestinal histology of blue discus / Influência de um “blend” de aminoácidos essenciais sobre o crescimento, atividade enzimática e histologia intestinal de acará disco

Symphysodon aequifasciatus is a fish with a disk-shaped body and bright colors, important characteristics of ornamental fish. We evaluated amino acid supplementation strategies to reduce crude protein in the diet for evaluation of performance, the content of digestive enzymes, liver metabolism, and intestinal histopathology. A total of 180 fish were randomly distributed in 12 separate 50 L glass aquariums, consisting of a completely randomized design with four treatments (DC - Control diet with 34.4% crude protein; DL - Control diet plus 1% of lysine; DEAA - Control diet plus 1% free essential amino acids (threonine, phenylalanine, leucine, valine, arginine, and tryptophan); and DHP - Diet with a high level of crude protein 48.4%), three repetitions, lasting 60 days. The use of DL and DEAA diets resulted in higher intestinal villus height and higher zootechnical performance. The use of DL diet increased alkaline phosphatase and digestive amylase activity. The use of DHP diets promotes severe liver changes due to increased activity of Alanine aminotraserase. Therefore, it was possible to observe that the use of amino acids can supply the nutritional need of blue discus. Supplementation of diets with AAs allows the reduction of dietary protein, which is a strategy for feeding management.(AU)

Symphysodon aequifasciatus é um peixe com corpo em forma de disco e cores brilhantes, características importantes dos peixes ornamentais. Avaliamos estratégias de suplementação de aminoácidos (AAs) para reduzir a proteína bruta na dieta com relação ao desempenho, atividade das enzimas digestivas, metabolismo hepático e histopatologia intestinal. Um total de 180 peixes foram distribuídos aleatoriamente em 12 aquários de vidro de 50 L, consistindo em um delineamento inteiramente casualizado, com quatro tratamentos (DC Dieta controle com 34,4% de proteina bruta; DL - Dieta controle acrescida de 1% de lisina; DEAA Dieta controle acrescida de 1% aminoácidos essenciais livres (treonina, fenilalanina, leucina, valina, arginina e triptofano) e DHP Dieta com alto nível de proteína bruta 48,4%), e três repetições, com duração de 60 dias. Os resultados demostraram que a utilização das dietas DL e DEAA refletiram em maior altura de vilosidade intestinal e maior desempenho zootécnico. A utilização de dieta DL promoveu aumento da atividade da fosfatase alcalina e amilase digestiva. A utilização de dietas DHP resultou em alterações hepáticas graves, devido a maior atividade da Alanina aminotransferase. Portanto, foi possível observar que a utilização de aminoácidos pode suprir a necessidade nutricional de acará disco. A suplementação de dietas com AAs permite a redução da proteína dietética, o que é uma estratégia para o manejo alimentar.(AU)