Production and essential oil quality of Varroniacurassavica DC.submitted to different spacing between plants, harvest season and drying temperatures of leaves / Produção e qualidade do óleo essencial deVarronia curassavicaDC. submetida à diferentes espaçamentos entre plantas, épocas de colheita e temperaturas de secagem das folhas

ABSTRACT: Varroniacurassavica is a Brazilian native medicinal species. Among the critical points influencing the phytochemical quality of bioactive compounds is the spacing between plants, harvest and post-harvest. This research aimed evaluated the influence of plant distance, harvest season, and leaves drying temperature on the yield and phytochemical quality of V. curassavica essential oil. The organic cultivation was carried out in 2018/2019 using 0.6 x 1.0; 0.8 x 1.0; 1.0 x 1.0; 1.0 x 1.6 m spacing between plants. The macro and micronutrient contents of the leaves were evaluated and no considerable changes were observed. In 2018 the harvest was performed in summer, autumn, and winter, and the harvested leaves were immediately submitted to the drying process at 40 °C. In 2019 the harvest was performed in winter, and the leaves were submitted to the drying process at 40, 50, and 60 °C. The essential oil was extracted by hydrodistillation and the chemical constituents were evaluated using CG-MS. The essential oil yield was significantly higher in winter and used 0.8 x 1.0 m and 1.0 x 1.0 m spacing between plants. The alpha-humulene content remained within the recommended standards at all analyzed temperatures. Although, the drying temperatures tested did not compromise the alpha-humulene content, the increasing temperature caused a reduction in the essential oil yield. Thus, it is recommended for the organic cultivation of V. currassavica the spacing of 0.8 x 1.0 m and 1.0 m x 1.0 m, and the drying of its leaves between 40 and 50 °C to earn the highest essential oil yield and phytochemical quality.

RESUMO: Varronia curassavica DC. é uma espécie medicinal nativa do Brasil. Dentre os pontos críticos que influenciam a qualidade fitoquímica dos compostos bioativos destacam-se o espaçamento entre plantas, a colheita e a pós-colheita. Este trabalho teve como objetivo avaliar a influência do espaçamento entre plantas, da época de colheita e da temperatura de secagem das folhas na produtividade e na qualidade fitoquímica do óleo essencial de V. curassavica. O cultivo orgânico foi realizado em 2018/2019 utilizando-se os seguintes espaçamentos entre plantas: 0.6 x 1.0; 0.8 x 1.0; 1.0 x 1.0; 1.0 x 1.6 m. Os teores de macro e micronutrientes das folhas foram avaliados e não se observou alterações consideráveis. Em 2018, as colheitas foram realizadas no verão, outono e inverno, sendo as folhas colhidas imediatamente submetidas ao processo de secagem a 40 °C. Em 2019, a colheita foi realizada no inverno, sendo as folhas submetidas ao processo de secagem a 40, 50 e 60 °C. O óleo essencial foi extraído por hidrodestilação e os constituintes químicos identificados CG-MS. A produção de óleo essencial foi significativamente maior no inverno e nos espaçamentos de 0,8 x 1,0 m e 1,0 x 1,0 m entre plantas. O teor de alfa-humuleno manteve-se dentro dos padrões recomendados em todas as temperaturas analisadas. Embora as temperaturas de secagem testadas não tenham comprometido o teor de alfa-humuleno, o aumento da temperatura ocasionou redução no rendimento do óleo essencial. Assim, recomenda-se o cultivo orgânico de V. currassavica no espaçamento entre 0,8 x 1,0 m e 1,0 m x 1,0 m, e a secagem de suas folhas entre 40 e 50 °C visando obter o maior rendimento e melhor qualidade fitoquímica de óleo essencial.

Production and essential oil quality of Varroniacurassavica DC. submitted to different spacing between plants, harvest season and drying temperatures of leaves / Produção e qualidade do óleo essencial deVarronia curassavicaDC. submetida à diferentes espaçamentos entre plantas, épocas de colheita e temperaturas de secagem das folhas

Varroniacurassavica is a Brazilian native medicinal species. Among the critical points influencing the phytochemical quality of bioactive compounds is the spacing between plants, harvest and post-harvest. This research aimed evaluated the influence of plant distance, harvest season, and leaves drying temperature on the yield and phytochemical quality of V. curassavica essential oil. The organic cultivation was carried out in 2018/2019 using 0.6 x 1.0; 0.8 x 1.0; 1.0 x 1.0; 1.0 x 1.6 m spacing between plants. The macro and micronutrient contents of the leaves were evaluated and no considerable changes were observed. In 2018 the harvest was performed in summer, autumn, and winter, and the harvested leaves were immediately submitted to the drying process at 40 °C. In 2019 the harvest was performed in winter, and the leaves were submitted to the drying process at 40, 50, and 60 °C. The essential oil was extracted by hydrodistillation and the chemical constituents were evaluated using CG-MS. The essential oil yield was significantly higher in winter and used 0.8 x 1.0 m and 1.0 x 1.0 m spacing between plants. The alpha-humulene content remained within the recommended standards at all analyzed temperatures. Although, the drying temperatures tested did not compromise the alpha-humulene content, the increasing temperature caused a reduction in the essential oil yield. Thus, it is recommended for the organic cultivation of V. currassavica the spacing of 0.8 x 1.0 m and 1.0 m x 1.0 m, and the drying of its leaves between 40 and 50 °C to earn the highest essential oil yield and phytochemical quality.

Varronia curassavica DC. é uma espécie medicinal nativa do Brasil. Dentre os pontos críticos que influenciam a qualidade fitoquímica dos compostos bioativos destacam-se o espaçamento entre plantas, a colheita e a pós-colheita. Este trabalho teve como objetivo avaliar a influência do espaçamento entre plantas, da época de colheita e da temperatura de secagem das folhas na produtividade e na qualidade fitoquímica do óleo essencial de V. curassavica. O cultivo orgânico foi realizado em 2018/2019 utilizando-se os seguintes espaçamentos entre plantas: 0.6 x 1.0; 0.8 x 1.0; 1.0 x 1.0; 1.0 x 1.6 m. Os teores de macro e micronutrientes das folhas foram avaliados e não se observou alterações consideráveis. Em 2018, as colheitas foram realizadas no verão, outono e inverno, sendo as folhas colhidas imediatamente submetidas ao processo de secagem a 40 °C. Em 2019, a colheita foi realizada no inverno, sendo as folhas submetidas ao processo de secagem a 40, 50 e 60 °C. O óleo essencial foi extraído por hidrodestilação e os constituintes químicos identificados CG-MS. A produção de óleo essencial foi significativamente maior no inverno e nos espaçamentos de 0,8 x 1,0 m e 1,0 x 1,0 m entre plantas. O teor de alfa-humuleno manteve-se dentro dos padrões recomendados em todas as temperaturas analisadas. Embora as temperaturas de secagem testadas não tenham comprometido o teor de alfa-humuleno, o aumento da temperatura ocasionou redução no rendimento do óleo essencial. Assim, recomenda-se o cultivo orgânico de V. currassavica no espaçamento entre 0,8 x 1,0 m e 1,0 m x 1,0 m, e a secagem de suas folhas entre 40 e 50 °C visando obter o maior rendimento e melhor qualidade fitoquímica de óleo essencial.

Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum

Starch processing industries use amylases, accounting for approximately 30% of the world's enzyme market. Previously, an amylase-producing strain of Epicoccum nigrumwas isolated from maize grains. Although E. nigrumamylase production is already reported in the literature, no published data on production optimization or characterization of the produced enzyme exists. The objectives of this work were to improve the amylase production by theE. nigrumPG 16 strain and to purify and characterize the produced enzyme. The E. nigrumPG 16 amylase production best conditions in submerged culture were: inoculum of 4% (v v-1) of a five-days-old stationary culture homogenate, agitation at 100 rpm, 25°C, natural light, 72 hours of incubation, starch as thecarbon source, and an initial medium pH of 7.0. A molecular exclusion chromatography profile has shown the production of only one amylase, which was partially purified with ammonium precipitation and dialysis. The enzyme optima pH and temperature are 6.0 and 50°C, respectively. The partially purified enzyme lostits activity when incubated for 30 min in temperatures above 40°C, presenting a T50of 46.25°C. The KMand Vmaxof the partially purified enzyme are 1.72 mg mL-1of starch and 0.15 mgmin-1of degraded starch, respectively. The ion Ca2+slightly activated the studied enzyme. The ions Cu2+, Zn2+, and Fe3+and the detergents SDS and Tween 80 acted as inhibitors of the studied enzyme. The partially purified enzyme released glucose from p-nitrophenyl α-D-glucopyranoside (p-NPG). Glucose was the enzyme's main product from starch hydrolysis, as evidenced by thin-layer chromatography. The E. nigrumPG 16 studied enzyme is a glucoamylase and represents an alternative for enzymatic starch hydrolysis.(AU)

Effect of clarification on some characteristics of pomegranate juice processed by two different methods during storagein Ali Ahmed / Efeito daclarificação sobre algumas características do suco de romã processado por dois métodos diferentes durante o armazenamento

This study aimed to assess the effectiveness of two clarifying procedures and their effects on some properties in thermally or microwave-pasteurized pomegranate juices. The experiment consisted in combining pectinase and protease as well as chitosan and gelatin once they were being storedin the refrigerator. The experiment consisted in three parts, being the first one a fresh juice without any clarification treatment, whereas, the second one was the fresh juice treated with clarifying agent consisted of pectinaseand protease mixture ratio (2:1) at 0.75 v/v,and 50°C for 20 min. Finally, the third one was fresh juice treated with chitosan and gelatin mixture at 0.4 and 0.8) g/L, respectively, at 50°C for (20) min. The pasteurization of all three experiments was done by using two techniques, i.e., one with thermal water bath at 85°C for twomin and the microwave at 400 Watts for twomin. All pomegranate juice bottles were stored at 4°C for threemonths. The results showed a significant effect of the clarificationmethod variable on the properties studied, especially turbidity, polyphenol and anthocyanin values. Moreover, the juice clarified with the enzymatic clarification method had better characteristics than the traditional ones during storage, what has therefore a better commercial appeal. The area of significance was founded with the use of traditional clarification with concentration at (0.4 and 0.8) g/l, and microwave pasteurization with 400 watts and zero month of storage at 4°C, respectively, which is provided a minimum turbidity value.(AU)

Este estudo teve como objetivo avaliar a efetividade de dois procedimentos esclarecedores e seus efeitos sobre algumas propriedades em sucos de romã termicamente ou pasteurizados por micro-ondas. O experimento consistiu em combinar pectinase e protease, bem como quitosana e gelatina, uma vez que estavam sendo armazenadas na geladeira. O experimento consistiu em três partes, sendo a primeira um suco fresco sem qualquer tratamento de clarificação, enquanto a segunda foi o suco fresco tratado com agente clarificante consistiu na proporção de mistura de pectinase e protease (2:1) a 0,75 v/v, e 50°C por 20 min. Finalmente, o terceiro foi suco fresco tratado com mistura de quitosana e gelatina a 0,4 e 0,8) g/L, respectivamente, a 50°C por (20) min. A pasteurização dos três experimentos foi realizada utilizando-se duas técnicas, ou seja, uma com banho de água termal a 85°C por doismin e a micro-ondas a 400 Watts por doismin. Todas as garrafasde suco de romã foram armazenadas a 4°C por trêsmeses. Os resultados mostraram um efeito significativo da variável método de clarificação sobre as propriedades estudadas, especialmente os valores de turbidez, polifenóis e antocianinas. Além disso, o suco clarificado com o método de clarificação enzimática apresentou melhores características do que os tradicionais durante o armazenamento, o que tem, portanto, um melhor apelo comercial. A área de significância foi fundada com o uso de clarificação tradicional com concentração a (0,4 e 0,8) g/l, e pasteurização por micro-ondas com 400 watts e zero mês de armazenamento a 4°C, respectivamente, o que proporciona um valor mínimo de turbidez.(AU)

The effect of olsalazine of chinese generic drugs on ulcerative colitis induced by dextran sulfate sodium salt in BALB/c mice

Purpose: To explore effect and mechanism of olsalazine of Chinese generic drugs on ulcerative colitis induced by dextran sulfate sodium salt (DSS) in BALB/c mice. Methods: The mouse model of ulcerative colitis was induced by free drinking of 3% (w/v) DSS aqueous solution for seven days. The mice were treated with olsalazine (0.6 g·kg-1) of Chinese generic drugs. The therapeutic effect of olsalazine on ulcerative colitis mice was evaluated by measuring disease activity index (DAI), colonic mucosal injury index (CMDI), histopathological score (HS), and detected the expression levels of interleukin (IL)-2, IL-10, tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), IL-1ß in serum and IL-7, IL-17, IL-22, epidermal growth factor (EGF), transforming growth factor ß1 (TGF-ß1) in colonic homogenate of mice. Results: Olsalazine significantly increased the contents of IL-2, IL-10, IL-22, TGF and EGF in ulcerative colitis rats, and significantly decreased the scores of DAI, CMDI, HS and the contents in IL-7, IL-17, TNF-α, IL-1ß and IFN-γ when compared with the model group. It improved the degree of colonic lesion in ulcerative colitis mice. Conclusions: It was suggested that olsalazine has a therapeutic effect on ulcerative colitis induced by DSS in mice, and the mechanism may be related to the increase of IL-2, IL-10, IL-22, TGF, and EGF and the decrease of the expression of IL-7, IL-17, TNF-α, IL-1ß, and IFN-γ.

The inhibitory and anticancer properties of Annona squamosa L. seed extracts / Propriedades inibitórias e anticancerígenas de extratos de sementes de Annona squamosa L

Although Annona squamosa Linn. (Annonaceae) has been used in traditional medicine and is known to have several pharmacological properties, its impact on EGFR kinase has not been fully investigated. An assay (biochemical) was used to govern the potential of different A. squamosa seed extracts to scavenge free radicals in petroleum ether, acetone, ethanol, and methanol. We also tested A. squamosa leaf extracts for their ability to inhibit the growth of HEK 293, MCF7, and HepG2 cell lines. The PSE, ASE, ESE, and MSE all contained anti-cancer substances like anethole, cyclopentane, 1,1,3-trimethyl, and phosphonate oxide tributyl, according to phytochemical analysis. ESE extracts from A. squamosa seeds have been selected based on free radical generation probabilities, cytotoxicity studies, and phytochemical analysis. Subsequent insilico studies have been conducted, and the results have shown that interactions between compounds present in ESE extracts and the EGFR kinase are what give these compounds their inhibitory effects. Preliminary phytochemical and pharmacological activities were studied and reported. A. squamosa ESE extracts inhibited the growth of MCF7 cells, and a pharmacokinetic study showed that the compounds anethole, cyclopentane, 1,1,3-trimethyl, and phosphonium oxide tributyl had few undesirable side effects. These substances can be used to both prevent and treat cancer diseases.

Embora a Annona squamosa Linn. (Annonaceae) tenha sido utilizada na medicina tradicional e seja conhecida por diversas propriedades farmacológicas, seu impacto na EGFR quinase ainda não foi totalmente investigado. Um ensaio bioquímico foi utilizado para controlar o potencial de diferentes extratos de sementes de A. squamosa para eliminar radicais livres em éter de petróleo, acetona, etanol e metanol. Extratos de folhas de A. squamosa também foram analisados em relação à sua capacidade de inibir o crescimento de linhagens celulares HEK 293, MCF7 e HepG2. O PSE, ASE, ESE e MSE continham substâncias anticancerígenas como anetol, ciclopentano, 1,1,3-trimetil e óxido de fosfonato tributil, de acordo com a análise fitoquímica. Extratos de ESE de sementes de A. squamosa foram selecionados com base em probabilidades de geração de radicais livres, estudos de citotoxicidade e análise fitoquímica. Estudos in silico subsequentes foram realizados e os resultados mostraram que as interações entre os compostos presentes nos extratos de ESE e a EGFR quinase são o que confere a esses compostos seus efeitos inibitórios. As atividades fitoquímicas e farmacológicas preliminares foram estudadas e relatadas. Os extratos de ESSE de A. squamosa inibiram o crescimento de células MCF7, e um estudo farmacocinético mostrou que os compostos anetol, ciclopentano, 1,1,3-trimetil e óxido de fosfônio tributil tiveram poucos efeitos colaterais indesejáveis. Essas substâncias podem ser usadas para prevenir e tratar doenças cancerígenas.

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.

Sperm cell capacitation status of ram semen after cooling

Background: The use of conventional artificial insemination (AI) in sheep production is usually associated with lower fertility rates when frozen semen is used. Cooled ram semen has been an alternative over frozen semen due to the higher viability, seminal quality and fertility rates following AI. The semen preservation process promotes sperm cell modifications similar to capacitation (capacitation-like) that causes cell damage affecting viability and seminal quality, but such effects are unclear for cooled semen. The aim of this study was to determine the status of sperm cell capacitation (CA) and acrosome reaction (AR) during ram semen processing and cooling under different extenders, dilution factors, and aerobiosis conditions as a function of storage time at 5o C. Materials, Methods & Results: Two consecutive ejaculates per day per male were collected from 2 adult rams by artificial vagina at 48-72 h intervals, in three replications. After macro- and microscopic evaluations, semen was segregated into groups under 3 extenders (Tris-egg yolk or TY, citrate-egg yolk or CY, skimmed milk or SM), 2 dilution factors (1 x 109 or Bi, 100 x 106 or Mi cells/mL), and 2 aerobiosis conditions (aerobic or A, semi-anaerobic or SA). Diluted semen was cooled to 5ºC and stored for up to 72 h, with evaluations every 24 h. Aliquots of fresh ejaculates and of each cooled diluted subgroup, according to extender, dilution, and aerobiosis, were collected at times T0 and T72 for determination of acrosome status and membrane integrity by the chlortetracycline (CTC) and trypan blue-Giemsa stainings, respectively. No differences were detected in sperm cell motility (M) and motility vigor (V) between fresh and diluted semen. After cooling, a significant decrease in M was observed after 48 h in CY and SM compared with fresh semen and 0 h of cooling, while V started to decrease after 24 h in CY compared with TY. Likewise, M/V from different dilutions and aerobic conditions decreased more significantly after 48 and 24 h of cooling, respectively. The sperm capacitation status did not show differences in the proportion of non-capacitated (NCA), CA and AR sperm cells between TY, CY, and SM extenders (NCA: 75.0%, 71.3%, 74.0%; CA: 15.7%, 17.2%, 15.9%; AR: 9.3%, 11.5%, 10.2%) or between Bi and Mi dilutions (NCA: 74.0%, 72.9%; CA: 15.9%, 16.6%; AR: 10.1%, 10.5%), respectively. However, differences (P < 0.05) were observed between A and SA aerobic conditions, with CA (17.0% vs. 15.5%) and AR (11.9% vs. 8.7%) rates being higher in A than SA, respectively, with no differences in NCA (71.1% vs. 75.8%), irrespective of the storage time. Sperm cell viability decreased after 48 h, especially in CY (P < 0.05). Discussion: Ram sperm cells can suffer irreversible damage due to thermal shock during cooling. Egg yolk-based extenders provide phospholipids and cholesterol to protect the sperm cell membrane during the thermal shock caused by the change in temperature. In this study, sperm cells had irreversible decreases in M/V, with increase in acrosome and plasma membrane damage after cooling to 5ºC. The largest and smallest decreases in M and V over time were observed in the CY and TY extenders, respectively. In addition to the extender type, the semen preservation method and storage time promoted changes in the capacitation status, AR and in sperm cell viability, which per se were associated with a decrease in semen fertility. In fact, the proportions of CA and/or AR sperm cells gradually increased over time after dilution and storage at 5ºC, with a negative correlation between sperm cell viability and M/V over time. In summary, extender and cooling time affected mostly M/V, while aerobiosis condition and dilution factor were more associated with acrosome status and sperm survival, with the extender having less impact on the acrosome status as a function of time.

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)

Isolation and characterization of five novel disulfide-poor conopeptides from Conus marmoreus venom

Background: Conopeptides from cone snail venom have aroused great interest related to the discovery of novel bioactive candidates, due to their excellent prospects for the treatment of various health problems such as pain, addiction, psychosis and epilepsy. In order to explore novel biopeptides, we investigated the structure and function of five novel conopeptides isolated from the venom of Conus marmoreus from South China Sea. Methods: C. marmoreus crude venom was prepared, fractionated and purified by HPLC system. The primary sequences of the five novel disulfide-poor conopeptides Mr-1 to Mr-5 were identified by comprehensive analysis of de novo MALDI-TOF tandem mass spectrometry and Edman degradation data. In order to investigate their function, these five conopeptides were synthesized by Fmoc-SPPS chemistry, and their biological effects at several heterologous rat nicotinic acetylcholine receptor (nAChR) subtypes (α1β1δε, α3β2, α3β4, α4β2) were determined by electrophysiological technique. Results: Five novel disulfide-poor conopeptides were identified and named as follows: Mr-1 (DWEYHAHPKPNSFWT), Mr-2 (YPTRAYPSNKFG), Mr-3 (NVIQAPAQSVAPP NTST), Mr-4 [KENVLNKLKSK(L/I)] and Mr-5 [NAVAAAN(L/I)PG(L/I)V]. None of them contains a disulfide bond. The sequences of conopeptides Mr-2 to Mr-5 do not belong to any category of the known disulfide-poor conopeptides. No significant activity against the above nAChR subtypes were observed for the five conopeptides at 100 µM. Conclusion: We purified and structurally characterized five novel disulfide-poor conopeptides from C. marmoreus crude venom and first investigated their nAChR inhibitory effects. This work expanded our knowledge on the structure and function of disulfide-poor conopeptides from this cone snail venom.(AU)

Effects of thermal environment on dairy cattle under a grazing system in the Western Amazon, Brazil / Efeitos do ambiente térmico em bovinos leiteiros em sistema de pastejo na Amazônia Ocidental, Brasil

This study aimed to evaluate the thermal conditions to which animals are exposed and their effect on the comfort/discomfort of dairy herds in a grazing system in the municipality of Sena Madureira, Acre State, Brazil. Eight farms and a total of 113 lactating crossbred cows were evaluated. Dry-bulb temperature (DBT, °C), relative humidity (RH, %), black globe temperature (BGT, °C), and wind speed (v, km h¹) were measured in January, February, and March (rainy season) to calculate temperature-humidity index (THI), black globe-humidity index (GTWBI), and radiant heat load (RHL). Moreover, the animals were measured for the physiological variables: rectal temperature (RT), heart rate (HR), and respiratory rate (RR). Significant differences (P<0.05) were observed for THI, GTWBI, and RHL. THI reached a mean value of 80 on farm G in January. Farm F had a GTWBI value of 79 in February. RHL reached the highest value (510 W m²). RT and HR showed no differences among the analyzed months. Thus, the environmental conditions in which dairy cows are housed at milking time rarely promoted thermal discomfort, with farm G being the most affected by THI.

Objetivou-se avaliar as condições térmicas a que os animais estão submetidos, seu efeito no conforto/desconforto de rebanhos leiteiros em sistema de pastejo, no município de Sena Madureira-Acre. Foram avaliadas oito propriedades e o total de 113 vacas mestiças em lactação. As medições ocorreram nos meses: janeiro, fevereiro e março (inverno amazônico), mensurando-se: temperatura do bulbo seco (TBS, °C), umidade relativa do ar (UR, %) e temperatura de globo negro (TGN, °C), velocidade do vento (v, km/h) para cálculos dos índices da temperatura e umidade (ITU), índice da temperatura de globo e umidade (ITGU) e carga térmica radiante (CTR), além da temperatura retal (TR), frequência cardíaca (FC), frequência respiratória (FR). Entre as propriedades analisadas, houve diferenças significativas (P<0,05) para ITU, ITGU e CTR. Destaca-se que o ITU na propriedade G, em janeiro, atingiu o valor médio de 80. A propriedade F, em fevereiro, teve valor de 79 para o ITGU. A CTR apresentou carga elevada (510 W.m³). A TR e a FC não apresentaram diferença entre os meses analisados. Concluiu-se que o ambiente ao qual as vacas leiteiras estão alojadas na hora da ordenha encontraram-se poucas vezes em situação de desconforto térmico, sendo a propriedade, a mais afetada pelo ITU.

Pantanals fish native meatballs has the nutritional values increased with the use of pequi in its recipe

The s tudy aimed to evaluate whether the addition of pequi increases the nutritional v alue of meatballs made with piranha and pacu fillets. Piranhas were obtained fro m the colony of fishermen; pacu with the fish farmer and the pequis were collected in nature. Fish were filleted and triturated, and the result ing masses were used to prepare 4 types of meatballs: piranha fillet with 2.0% pequi pulp, piranha fillet without pequi pulp, pacu fillet with w ith 2.0% pequi pulp, and pacu fillet without pequipulp. After preparation, accep tance, purchase intention and frequency of consumption were evaluated with untra ined tasters. Chemical composition parameters evaluated were crude protein, lipi ds, ash, moisture, carotenoids and antioxidant activity. Results of acce ptability for all formulations were similar. The chemical composit ion of piranha meatballs presented differences (p<0.05) for lipids and carotenoi ds. Pacu meatballs showed no difference (p>0.05) for moisture, carotenoids,and a ntioxidant activity. The addition of pequi did not alter the organoleptic c haracteristics, however increased the nutritional values, therefore, it can add value to the product to be marketed and be more nutritionally attractive to the consumer.(AU)

Genetic diversity in Amazonian Jundiá (Leiarius marmoratus) stocks using he erologous primers

With the rise of world fish farming, the national scenario is favorable for using native fish for intensive farming. Among the catfish, the Amazonian Jundiá (Leiarius marmoratus) is a robust candidate, easy to grow and with good organoleptic characteristics in its flesh. For productive success in c aptivity, it is necessary to consider some questions about the species, such as genetic variability, which must have an acceptable level in a breeding stoc k, in order to maintain a good diversity; this reduces losses due to inbreeding and low diversity. Therefore, the objective of this study was to characterize the genetic variability of commercial stocks of L. marmoratusfrom the State of Mato Grosso through microsatellite molecular markers. We analyzed 143 individuals from three stocks. The mean hete rozygosity and the inbreeding coefficients observed were 0.060; 0.084; 0 .141; and 0.539; 0.562; 0.514, respectively, for the stocks of Campo V erde, Juína,and Nova Mutum. The Deviation in the Hardy-Weinberg equilib rium was observed in most of the lociin the three populations. Considering the genetic differentiation, it is concluded that the three populations are very clo se genetically, which requires introduction of new genetic material in the stock s to enrich them genetically for a later reproductive program.(AU)

Determination of the Relationship of Serum Amino Acid Profile with Sex and Body Weight in Healthy Geese by Liquid Chromatography-Tandem Mass Spectrometry

The aim of this study is to determine the serum amino acid profile using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS / MS) in healthy male and female geese of the same age that were raised in similar care and feeding conditions. The animal material of the study consisted of a total of 41 geese, 18 male, and 23 female of the same age (9 months). After a 12h fasting period of the geese, blood samples were taken from V. subcutanaeulnaris into tubes without anticoagulant. After separating the serums, the samples were preserved at -20 O C degrees until Methylglutaryl (Met-Glu), Valine (Val), Leucyl-Isoleucine (Leu-Ile), Methionine (Met), Phenylalanine (Phe), Argininosuccinate (ASA), Tyrosine (Tyr), Aspartic acid (Asp), Alanine (Ala), Arginine (Arg), Citrulline (Cit), Glycine (Gly), Ornithine (Orn), Glutamic acid (Glu) were analyzed. The Val, Asp, Arg, Cit, Gly, and Orn levels of male geese were higher compared to female geese in the research (p<0.05). It was determined that Asp, Arg, Cit, and Gly levels increased as the body weights of the geese increased (p<0.05). It was also determined that the effect of Gender x Body Weight interaction on Val, Cit, and Orn levels was significant (p<0.05). As a result; it has been concluded that the serum amino acid profile of healthy geese can vary according to gender and live weight, and more studies are needed to elucidate the reasons for these changes.(AU)

Digestive system of Anourosorex squamipes - appearance and morphological features

Background: Anourosorex squamipes have a wide range of feeding habits, which is a kind of omnivorous animal. As an indispensable part of organisms, the differentiation degree of digestive system indicates the evolution degree of species. The higher the evolution degree of animals, the higher the differentiation of digestive system. At present, the research on Anourosorex squamipes digestive system mainly focuses on its feeding habits and mainly depends on the direct observation of gastric contents with the naked eye, but the research on the morphology of digestive system has not been reported. Materials, Methods & Results: Twenty adult Anourosorex squamipes were selected to investigate the characteristics of the appearance, including the contour parameters (body weight, body length, forefoot length, posterior foot length, nasal length, tail length), digestive system parameters (length of esophagus, intestinal length, intestinal weight, stomach weight and liver weight) and morphological structure of the small intestine by histological methods. The results showed that there were no significant differences in body weight, body length, forefoot length, posterior foot length, nasal length and tail length between males and females in Anourosorex squamipes, and no significant differences were observed in parameters of digestive system in length of esophagus, intestinal length, intestinal weight, stomach weight and liver weight. There were no significant differences in villus height, muscular layer thickness and villus height/recess depth (V/C) between males and females, but only ileal crypt depth decreased significantly in males. Discussion: Animal morphological characteristics are closely related to their life habits, unique organs and lifestyles ensure that they thrive on earth. In our study, we found that unique characters are found by observing the appearance of Anourosorex squamipes, the snout is blunt and short, the eyes are degenerated, and the tail is very short and covered with scales, which may be related to its feeding habits, the living environment-long-term nocturnal life in underground with minimal exposure to sunlight, and the unique tail can reduce the friction between tail and soil during the movement. There are certain differences in the appearance parameters of male and female Anourosorex squamipes, but are not significant. The esophagus, stomach, intestine, liver and other parameters of Anourosorex squamipes have little difference between males and females, indicating that Anourosorex squamipes don't have obvious distinctions between males and females in food intake. However, the liver weight of male Anourosorex squamipes is larger than female, indicating that the physiological metabolic capacity of male is larger than female. It is speculated that this may be related to the difference in the amount of activity between males and females in daily life. Males are more active and require more energy, but their digestion and absorption abilities are weaker than females. Therefore, the male Anourosorex squamipes needs more food to provide energy to meet the daily physiological metabolism, and the larger stomach capacity provides the essential conditions for the male to store more food. In addition, we found that the crypt depth of male Anourosorex squamipes is generally greater than that of females, especially in the ileal crypt depth, indicating that the female's cell maturation rate is greater than that of the male. It is speculated that the female has a greater demand for nutrient absorption and better digestion and absorption of food, because females need to give birth to offspring and require greater nutrition.

Chemical composition and antioxidant activity of leaf essential oil from Calyptranthes concinna DC. (Myrtaceae)

The biodiversity found in Brazilian's ecosystems brings the possibility of discovering new natural products with wide application potentials. However, knowing their availability and chemical composition is crucial. Thus, the aim of this study was to evaluate the extraction yield, chemical composition and antioxidant activity of essential oil from fresh leaves of Calyptranthes concinnaDC., a native species of Myrtaceae occurring in Brazilian Atlantic Rain Forest. Plant samples were collected in SoutheasternBrazil and the essential oil was extracted by hydrodistillation. The chemical composition was evaluated by Gas Chromatography associated with Mass Spectrometry and antioxidant activity was measured using ABTS, DPPH and FRAP methods. The extraction yield obtained was 0.015% (v), and the chemical composition revealed elemicin, a phenylpropanoid as the major component (36.46%). Still, ß-caryophyllene (16.94%), germacrene B (8.28%) and spathulenol (7.33%) proved to be relevant for the same essential oil. Antioxidant activity was obtained for ABTS and DPPH radical scavenge (134.82 ± 2,9 and 93.70 ± 1.7 µM TE mL-1, respectively) and FRAP (11.31 ± 0.2 µM FeSO4mL-1OE), revealing hydrogen-donation as the main antioxidant mechanism. To our knowledge, this is the first report of antioxidant activity of C. concinna essential oil. The product presented compounds of great relevance, with possibilities of application in different areas including food, agriculture and pharmaceutical segments(AU)

Comparative characterization of digestive proteases in redhead cichlid (Vieja melanurus) and twoband cichlid (Vieja bifasciata) (Percoidei: Cichlidae)

In the Southeast of Mexico, there are many native cichlids with commercial interest such as redhead cichlid (Vieja melanurus) and twoband cichlid (V. bifasciata), which have a great local demand and excellent meat quality. However, it is necessary to implement their culture based on nutrition studies and digestive biochemistry. This study's objective was to characterize these two cichlids' digestive proteases (pH, temperature, and inhibitors) through biochemistry techniques. Results showed that V. melanurus and V. bifasciata have a digestive capacity analogous to other omnivore fishes, where the optimal pH values of stomach proteases (4 and 2, respectively) and intestinal proteases (6 and 12, respectively), the optimal temperature of acid (35°C and 55°C, respectively) and alkaline proteases (45°C and 55°C, respectively) are quite similar. Both species presented high thermal and pH stabilities. Inhibition showed that V. melanurus is more sensitive to specific inhibitors for alkaline proteases than V. bifasciata. In conclusion, V. bisfasciata and V. melanurus have different digestive protease patterns. Both species can hydrolyze different protein ingredients to formulate a specific diet. Nevertheless, V. bifasciata is more resistant to the presence of inhibitors, which allow it to include vegetable proteins in its diet.(AU)

En el sureste de México, existen muchas especies de cíclidos nativos de interés comercial como el cíclido rojo (Vieja melanurus) y el cíclido de dos bandas (V. bifasciata), los cuales tienen una gran demanda local y tienen una excelente calidad de carne; sin embargo, es necesario implementar su cultivo con base en estudios de nutrición y bioquímica digestiva. El objetivo de este estudio fue caracterizar las proteasas digestivas (pH, temperatura e inhibidores) de estos dos cíclidos nativos mediante técnicas bioquímicas. Los resultados mostraron que V. melanurus y V. bifasciata tienen una capacidad digestiva similar a otros peces omnívoros, donde los valores óptimos de pH de proteasas estomacales (4 y 2, respectivamente) e intestinales (6 y 12, respectivamente), la temperatura óptima de proteasas ácidas (35°C y 55°C, respectivamente) y alcalinas (45°C y 55°C, respectivamente) son muy parecidas. Ambas especies presentaron alta estabilidad térmica y de pH. La inhibición mostró que V. melanurus es más sensible a inhibidores específicos de proteasas alcalinas que V. bifasciata. En conclusión, V. bisfasciata y V. melanurus tienen diferentes patrones de proteasas digestivas, pero ambas especies pueden hidrolizar diversos ingredientes proteicos para formular dietas específicas; sin embargo, V. bifasciata es más resistente a la presencia de inhibidores, lo que permitiría incluir proteínas vegetales en su dieta(AU)

Comparative characterization of digestive proteases in redhead cichlid (Vieja melanurus) and twoband cichlid (Vieja bifasciata) (Percoidei: Cichlidae)

In the Southeast of Mexico, there are many native cichlids with commercial interest such as redhead cichlid (Vieja melanurus) and twoband cichlid (V. bifasciata), which have a great local demand and excellent meat quality. However, it is necessary to implement their culture based on nutrition studies and digestive biochemistry. This study's objective was to characterize these two cichlids' digestive proteases (pH, temperature, and inhibitors) through biochemistry techniques. Results showed that V. melanurus and V. bifasciata have a digestive capacity analogous to other omnivore fishes, where the optimal pH values of stomach proteases (4 and 2, respectively) and intestinal proteases (6 and 12, respectively), the optimal temperature of acid (35°C and 55°C, respectively) and alkaline proteases (45°C and 55°C, respectively) are quite similar. Both species presented high thermal and pH stabilities. Inhibition showed that V. melanurus is more sensitive to specific inhibitors for alkaline proteases than V. bifasciata. In conclusion, V. bisfasciata and V. melanurus have different digestive protease patterns. Both species can hydrolyze different protein ingredients to formulate a specific diet. Nevertheless, V. bifasciata is more resistant to the presence of inhibitors, which allow it to include vegetable proteins in its diet.(AU)

En el sureste de México, existen muchas especies de cíclidos nativos de interés comercial como el cíclido rojo (Vieja melanurus) y el cíclido de dos bandas (V. bifasciata), los cuales tienen una gran demanda local y tienen una excelente calidad de carne; sin embargo, es necesario implementar su cultivo con base en estudios de nutrición y bioquímica digestiva. El objetivo de este estudio fue caracterizar las proteasas digestivas (pH, temperatura e inhibidores) de estos dos cíclidos nativos mediante técnicas bioquímicas. Los resultados mostraron que V. melanurus y V. bifasciata tienen una capacidad digestiva similar a otros peces omnívoros, donde los valores óptimos de pH de proteasas estomacales (4 y 2, respectivamente) e intestinales (6 y 12, respectivamente), la temperatura óptima de proteasas ácidas (35°C y 55°C, respectivamente) y alcalinas (45°C y 55°C, respectivamente) son muy parecidas. Ambas especies presentaron alta estabilidad térmica y de pH. La inhibición mostró que V. melanurus es más sensible a inhibidores específicos de proteasas alcalinas que V. bifasciata. En conclusión, V. bisfasciata y V. melanurus tienen diferentes patrones de proteasas digestivas, pero ambas especies pueden hidrolizar diversos ingredientes proteicos para formular dietas específicas; sin embargo, V. bifasciata es más resistente a la presencia de inhibidores, lo que permitiría incluir proteínas vegetales en su dieta.(AU)