Reduction in the in vitro ruminal methane production with the inclusion of Cucurbita foetidissima / Reducción en la producción ruminal de metano in vitro con la inclusión de Cucurbita foetidissima

ABSTRACT Objective. This research aimed to evaluate the inclusion of Cucurbita foetidissima (BG) leaves as a partial or total substitution of alfalfa hay (AH) in beef cattle diets on in vitro methane output, gas kinetics and volatile fatty acids production. Materials and Methods. Five experimental treatments were formulated with the inclusion of BG as alfalfa hay (AH) substitute at 0, 25, 50, 75 and 100% in experimental treatments denominated CON (control), BG25, BG50, BG75 and BG100, respectively. Results. Lignin and organic matter decreased with BG inclusion (p<0.05). However, NDF, CT and TPC increased with higher levels of BG (p<0.05). Maximum gas production diminished with BG inclusion (p<0.05); whereas, microbial protein production, specific gas production rate and latency period were not affected (p>0.05). Methane production decreased linearly with BG inclusion (p<0.05). Nevertheless, CO2 production showed no changes with inclusion of graded levels of BG in the experimental treatments (p>0.05). Additionally, acetate and butyrate were not affected by BG inclusion (p>0.05); although, propionate increased linearly among treatments (p<0.05). Furthermore, inclusion of 75% of BG reduced 31% in vitro methane production without changes in CO2 production, suggesting an inhibition of endogenous methanogenesis. Conclusions. These results promote the inclusion of BG leaves as a potential and environmentally friendly forage source for beef cattle feeding.

RESUMEN Objetivo. El objetivo de este estudio fue evaluar la inclusión de hojas de Cucurbita foetidissima (BG) como sustituto parcial o total de la alfafa (AH) en la dieta de ganado de engorda sobre la cinética de producción de gas y producción ruminal in vitro de metano y ácidos grasos volátiles. Materiales y Métodos. Se formularon cinco dietas con la inclusión de BG como sustituto de heno de alfalfa a 0, 25, 50, 75 y 100% en los tratamientos experimentales que se denominaron CON (control), BG25, BG50, BG75 y BG100, respectivamente. Resultados. La lignina y la materia orgánica disminuyeron con la inclusión de BG (p<0.05). La máxima producción de gas disminuyó con la inclusión de BG (p<0.05); mientras que la producción de proteína microbiana, la tasa específica de producción de gas y el tiempo de latencia no mostraron cambios(p>0.05). La producción de metano disminuyó linealmente con la inclusión de BG (p<0.05). No obstante, la producción de CO2 no mostró cambios con la inclusión de BG en los tratamientos (p<0.05). Adicionalmente, el acetato y el butirato no fueron afectados por la inclusión de BG (p>0.05); a pesar de que el propionato se incrementó linealmente (p<0.05). Además, la inclusión de BG en un 75% redujo la producción de metano 31% sin afectar la producción de CO2, lo que sugiere una inhibición de la metanogenesis endógena. Conclusiones. Estos resultados promueven la inclusión de hojas de BG como una fuente de forraje potencial y amigable con el ambiente en la alimentación de ganado de engorda.

Investigation on in Vitro Antioxidant Activity and Chemical Composition of Different Polar Parts of Extract of Rosa cymosa Roots / 中国实验方剂学杂志

Objective:To investigate the antioxidant activity and chemical composition of 75% ethanol extract of <italic>Rosa cymosa</italic> roots and its different polar parts. Method:The 75% ethanol extract of <italic>R. cymosa</italic> roots was divided into dichloromethane， ethyl acetate， <italic>n</italic>-butanol and water parts by organic solvent extraction. <italic>In vitro</italic> antioxidant activity of each fraction was determined by 1，1-diphenyl-2-picrylhydrazyl （DPPH） and 2，2'-azino-bis（3-ethylbenzothiazoline-6-sulfonic acid） （ABTS） free radical scavenging assays， as well as ferric reducing antioxidant power （FRAP） test. The contents of total triterpenes， total phenols， total tannins and condensed tannins in each fraction were determined by spectrophotometry. SPSS 24.0 software was used to conduct Pearson correlation analysis between the antioxidant activity of each fraction and the content of the main components， and then the main active fraction and the main active components were determined. The chemical constituents of the active fraction was analyzed by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MS/MS）， and the structures of the main chromatographic peaks were predicted. Result:Each fraction of <italic>R. cymosa</italic> roots had certain antioxidant activity， and there was a significant dose-effect relationship within a certain concentration range， but the antioxidant activity of different polar parts was different. In DPPH and ABTS free radical scavenging tests， the antioxidant activity of each fraction and vitamin C （VC， the positive drug） was ranked as ethyl acetate fraction>VC><italic>n</italic>-butanol fraction>ethanol extract>water fraction>dichloromethane fraction. In FRAP test， the activity of ethyl acetate fraction was weaker than that of VC， and the other order was unchanged. The contents of total triterpenes， total phenols， total tannins and condensed tannins in ethyl acetate fraction were 3.81%， 50.33%， 3.32%， and 39.79%， in <italic>n</italic>-butanol fraction were 0.88%， 41.42%， 2.25% and 23.55%， in ethanol extract were 2.90%， 41.95%， 3.43% and 20.14%， in water fraction were 0， 26.80%， 16.90% and 7.57%， and in dichloromethane fraction were 21.23%， 12.90%， 1.59%， and 6.17%， respectively. Correlation analysis results showed that the contents of total phenols and condensed tannins were positively correlated with the antioxidant activity， the contents of total triterpenes were negatively correlated with the antioxidant activity， and the correlation between total tannins and antioxidant activity was not obvious. A total of 26 compounds were identified from the ethyl acetate fraction by UPLC-Q-TOF-MS/MS， including 11 condensed tannins， 4 hydrolysable tannins， 6 triterpenes， 3 flavonoids， 1 benzoic acid derivative and 1 chlorogenic acid analogue. Conclusion:Ethyl acetate fraction is the main antioxidant active site of <italic>R. cymosa</italic> roots， and phenols mainly composed of condensed tannins are the main active components. The results can provide experimental basis for the development of natural antioxidants.

Nutrient Composition, Metabolites and Microbial Counts in Litter Material of Broiler Chicken Fed Condensed Tannins Containing Guava Leaf Meal Supplemented Diets: Reduce Environmental Pollution and Alternate Feedstuffs for Ruminants

Present study was conducted in 240 day-old broiler chicks to assess the effect of condensed tannins (CT) containing guava leaf meal (GLM) supplemented diet on nutrient composition, litter metabolites viz. pH, moisture and ammonia concentration) and microbial counts after completion of feeding trial of 42 days. Chicks were randomly allocated into 4 dietary treatments, with 4 replicates of 15 chicks in each pen in a completely randomized block design (CRD). Chicks were fed basal diets supplemented with GLM @ 1.5, 2.5 and 3.5 percent, in T1, T2 and T3 groups, respectively, while, T0 group fed only basal diet. Litter materials of chicks showed good nutrient profiles and properly sterilized litter material act as alternative feed ingredient for ruminants feed. Supplementation of GLM in the diets of chicks significantly (P<0.05) reduced moisture, pH, NH3 concentration, harmful Eschericia coli and Clostridia spp. counts in the litter materials and reduced environmental pollution. The litter material of GLM supplemented (T1, T2 and T3) chicken showed significantly higher (P<0.05) CP contents as compared to un-supplemented control. It can be concluded that properly sterilized litter material of GLM supplemented chicks act as cheaper alternative feedstuffs for ruminant’s ration and reduce environmental pollution by lowering NH3 concentration, harmful E. coli and Clostridia counts in broiler litter material

Evaluación de variables agronómicas, calidad del forraje y contenido de taninos condensados de la leguminosa Lotus corniculatus en respuesta a biofertilizante y fertilización química en condiciones agroecológicas de trópico alto andino colombiano

En la sabana de Bogotá, 2600 m.s.n.m., se analizaron variables agronómicas; altura de planta, número y longitud de ramas, producción de materia seca total ha-1, y variables de calidad del forraje, digestibilidad de materia seca, proteína cruda, extracto etéreo, fibra detergente ácido, fibra detergente neutra, calcio, así como el contenido de taninos condensados de la leguminosa Lotus corniculatus, mediante un diseño completo al azar de cuatro tratamientos aplicados al suelo Tl: sin fertilizante; T2: biofertilizante; T3: fertilización química, y T4: biofertilizante más fertilización química, con tres repeticiones por tratamiento. Se utilizó análisis de varianza (ANOVA) y prueba de comparación de medias de Tukey al 0,1%. Los resultados señalan que el cultivo de L corniculatus, responde muy bien a la biofertilización, ya que el T2 exhibió el mejor comportamiento para la mayoría de variables agronómicas y de calidad del forraje, mientras que T4 mostró los mejores resultados para número de ramas y menos contenido de taninos condensados. En digestibilidad de la materia seca T2 y T4 ostentaron un buen comportamiento con diferencias altamente significativas con respecto a TI y T3; en contenidos de grasa y calcio, el mejor comportamiento se presentó en el T3 y TI con diferencias altamente significativas. Se concluye que la biofertilización incide de manera favorable en las características edáficas, producción y calidad del forraje Lotus corniculatus, con mejores comportamientos en las principales variables evaluadas en el estudio.

In the savannah of Bogotá, 2600 m.s.n.m., were analyzed Agronomic variables; plant height, number and length of branches, total dry matter yield ha-1, and quality variables of forage, dry matter digestibility, raw protein, ethereal extract, Fiber Acid Detergent, Neutral Detergent Fiber calcium, as well as the condensed tannin content of the legume Lotus corniculatus, by means of a complete random design of four treatments applied to the soil Tl: no fertilizer; T2: biofertilizer; T3: chemical fertilization, and T4: biofertilizer with chemical fertilization, with three replicates per treatment. It was used analysis of variance (ANOVA) and comparison test ofTukey averages at 0.1%. The results indicate that the cultivation of L corniculatus responds very well to biofertilization because T2 exhibited the best performance for most changeful agronomic and forage quality while T4 showed the best results for number of branches and less content of condensed tannins. In dry matter digestibility T2 and T4 showed good behavior with highly significant differences with respect to Tl and T3; In fat and calcium contents, the best behavior was presented in T3 and Tl with highly significant differences. It is concluded that biofertilization favorably influences the edaphic characteristics, production and quality of the forage Lotus corniculatus, with better behaviors in the main variables evaluated in the study

Na planície de Bogotá, 2600 m.s.n.m., variáveis agronômicas analisadas; altura das plantas, número e comprimento das ramificações, a produção de matéria seca total ha-1, e digestibilidade qualidade da forragem variável de matéria seca, proteína bruta, extrato etéreo, fibra em detergente ácido, neutro fibra em detergente, de cálcio, e o taninos condensados conteúdo Lotus corniculatus leguminosa de, utilizando um desenho de blocos completos ao acaso, quatro tratamentos aplicados ao solo T1: não fertilizados; T2: biofertilizante; T3: adubação química, e T4: biofertilizantes fertilização mais química, com três réplicas por tratamento. análise de variância (ANOVA) e teste de Tukey comparação foi utilizada a 0,1%. Os resultados indicam que a cultura de L. corniculatus, responde muito bem ao Biofertilização como o T2 exibiu o melhor desempenho para a maioria das características agronômicas e qualidade da forragem, enquanto T4 apresentaram os melhores resultados para número de agências emenor teor de taninos condensados. Na seca T2 de digestibilidade e T4 que ostentava um bom desempenho com diferenças altamente significativas em relação ao T1 e T3; em gordura e teor de cálcio, o melhor desempenho foi apresentado nas diferenças altamente significativas T3 e T1. Conclui-se que Biofertilização favoravelmente impactos sobre as características do solo, produção e qualidade de forragem Lotus corniculatus, com melhores performances nas principais variáveis avaliadas no estudo.

Quality Standard for Yuhuang Sprays / 中国药师

Objective: To establish the quality standard for Yuhuang sprays. Methods: Berberine hydrochloride in Yuhuang sprays was qualitatively identified by TLC and determined by UV. The Vanillin-H2 SO4 method was employed for the determination of condensed tannins. Results:The spots obtained from the sample solutions showed the same color as those from the reference substance solution without interference from the negative sample. The calibration curve of berberiny hydrochloride was linear within the range of 1. 13-13. 60μg·ml-1(r=0. 999 9), and the average recovery was 99. 68% with RSD of 1. 53%(n=9). The calibration of catechin was linear within the range of 0. 03-0. 30 mg· ml-1(r=0. 999 3), and the average recovery was 99. 6% with RSD of 1. 76%(n=9). Conclusion:The method can be used in the quality control of Yuhuang sprays.

Analysis of condensed tannins' constituent units by thiolysis with cysteamine hydrochloride and analysis of thiolysis products' HPLC behavior / 国际药学研究杂志

Objective To establish a method of analyzing the structural units of condensed tannins from Chinese patent medicine Wei MaiNing. Methods The ethanol extract of Wei MaiNing capsule was depolymerized in the presence of cysteamine and analyzed by HPLC, and eight major thiolysis products were identified by LC-MS analysis combined with reference sample. Meanwhile, the influence of mobile phases with different pH values on the HPLC behavior of thiolysis products was investgated. Results Eight major thiolysis products were 4β-(2-aminoethylthio)catechin, 4β-(2-aminoethylthio)epicatechin, catechin, 4β-(2-aminoethylthio)epicatechin-3-O-gallate, epicatechin, 4β-(2-aminoethylthio)catechin-3-O-gallate, epicatechin-3-O-gallate and catechin-3-O-gallate. Variation of the pH value had more impact on the retention time of catechins' aminomethylthio derivatives than that of catechins', which led to the changes of chromatographic peaks order. Conclusion The method is simple, reliable, and could be used to determine the structural units of condensed tannins. In order to ensure the stability of derivatives retention time, pH values of mobile phases should be controlled strictly when thiolysis products are analyzed by HPLC.

Polifenoles y actividad antioxidante en Equisetum giganteum L. (Equisetaceae) / Polyphenols and antioxidant activity in Equisetum giganteum L. (Equisetaceae)

Equisetum giganteum L. (Equisetaceae) is a species native to South America and Central America, which is used in traditional medicine as a diuretic and in treating of various diseases. The aim of this paper was to study the polyphenol profile and antioxidant activity of extracts obtained from lateral branches and main stem of the plants. Quali- and quantitative differences were detected with higher contents of phenols, tannins, condensed tannins, flavonoids and hydroxicinnamic acids in the lateral branches. The proanthocyanidins propelargonidin and procyanidin only were detected in lateral branches. These extracts showed the highest antioxidant activity. Due to their higher concentration and chemical composition, lateral branches could be considered as a sort of source of compounds that would act as antioxidant.

Equisetum giganteum L. (Equisetaceae) es una especie nativa de Sudamérica y América Central, que es empleada en la medicina tradicional como diurético y en el tratamiento de diversas patologías. El objetivo de este trabajo fue estudiar el perfil de polifenoles y la actividad antioxidante de extractos obtenidos de las ramas laterales (tallos finos), tallos entrenudos y tallos basales. Se detectaron diferencias cuali-cuantitativas, con mayor contenido de fenoles, taninos, taninos condensados, flavonoides y ácidos hidroxicinámicos en las ramas laterales. Las proantocianidinas propelargonidina y procianidina sólo se detectaron en las ramas laterales. En estos extractos se detectó la mayor actividad antioxidante. Debido a la alta concentración y composición química, los tallos laterales podrían ser empleados como fuente de compuestos con actividad antioxidante.

Efeito de frações tânicas sobre parâmetros biológicos e nutricionais de Spodoptera frugiperda (Lepidoptera: Noctuidae) / Effect of tanical fractions on biological and nutritional parameters of Spodoptera frugiperda (Lepidoptera: Noctuidae)

Conduziu-se este trabalho, com o objetivo de quantificar taninos condensados presentes nas cascas do caule de Ochroma pyramidalis (Bombacaceae), Schinus terebinthifolius (Anacardiaceae), Enterolobium contortisiliquum (Fabaceae) e Peltophorum dubium (Fabaceae) e verificar o efeito de frações tânicas dessas espécies sobre parâmetros biológicos e nutricionais de S. frugiperda. A partir das cascas das espécies vegetais se prepararam extratos, dos quais alíquotas foram tomadas para a quantificação dos taninos. Em seguida, os extratos foram submetidos a fracionamento para a obtenção das frações tânicas, que foram incorporadas à dieta artificial e oferecidas para lagartas de S. frugiperda. Todas as espécies vegetais estudadas apresentaram taninos condensados, sendo que S. terebinthifolius apresentou menor quantidade desses compostos. A fração tânica de P. dubium causou redução no peso das lagartas e pupas, diminuição na sobrevivência de S. frugiperda durante as fases larval e pupal, e acarretou menor consumo alimentar, além de prolongar a duração das fases imaturas desse inseto. O tratamento com S. terebinthifolius causou redução na percentagem de sobrevivência, durante a fase de pupa. Pupas provenientes de lagartas que receberam tratamento com a fração de E. contortisiliquum tiveram aumento no seu período de desenvolvimento. Lagartas alimentadas com a fração proveniente de P. dubium apresentaram a menor taxa de crescimento relativo (RGR), menor eficiência de conversão do alimento digerido (ECD) e maior custo metabólico (CM). A digestibilidade aproximada (AD) foi maior para os tratamentos com as frações tânicas de P. dubium e S. terebinthifolius.

The objective of this study was to quantify condensed tannins present in the stem bark of Ochroma pyramidalis (Bombacaceae), Schinus terebinthifolius (Anacardiaceae), Enterolobium contortisiliquum (Fabaceae) and Peltophorum dubium (Fabaceae) and to verify the effect of tannic fractions of these species on biological and nutritional parameters of S. frugiperda. Extracts were prepared from the barks of the plant species and aliquots were taken from them to quantify the tannins. The extracts were then submitted to fractionation to obtain the tannic fractions, which ones were incorporated into an artificial diet and offered to the larvae of S. frugiperda. All the studied species showed condensed tannins, and S. terebinthifolius showed the lowest amount of these compounds. The tannin fraction of P. dubium caused a reduction in the weight of larvae and pupae, decreased the survival of S. frugiperda during the larval and pupal stages and led to a reduction of intake, beyond to extend the duration of immature stages of this insect. The treatment with S. terebinthifolius caused a reduction in the survival percentage during the pupal stage. Pupae from larvae that received the treatment with the fraction of E. contortisiliquum had an increase in its development period stage. Larvae fed with the fraction from P. dubium showed the lowest relative growth rate (RGR), lower efficiency of conversion of ingested food (ECI) and a higher metabolic cost (CM). The approximate digestibility (AD) was higher for the treatments with tannic fractions of P. dubium and S. terebinthifolius.

Antioxidant capacity of extracts and isolated compounds from Stryphnodendron obovatum Benth / Capacidade antioxidante de extratos e compostos isolados de Stryphnodendron obovatum Benth

The extract from stem bark of Stryphnodendron obovatum Benth. was chromatographed on a Sephadex® LH-20 column, and yielded nine compounds: gallic acid (GA), p-hydroxybenzoic acid (PHB), gallocatechin (GC), epigallocatechin (EPG), 4'-O-methylgallocatechin (MGC), epigallocatechin-(4β→8)-epigallocatechin (EPEP), epigallocatechin-(4β→8)-gallocatechin (EPGC), robinetinidol-(4α→8)-gallocatechin (ROGC) and robinetinidol-(4β→8)-epigallocatechin (ROEP). Evaluation of the antioxidant capacity in vitro by the methods of DPPH free radical (IC50; μg/mL) and reduction of the phosphomolybdenum complex (RAC) gave the following results, respectively: crude extract 4.52 and 0.8242; ethyl-acetate fraction 4.04 and 0.9537; aqueous fraction 5.58 and 0.9275. The crude extract and ethyl-acetate fraction were shown to possess an antioxidant capacity comparable to that of vitamin C (4.93 and 1.0). The values obtained by the DPPH free-radical method for the isolated compounds were IC50 (μM): GA=8.89; PHB=10.12; GC=16.46; EPG=13.20; MGC=21.00; EPEP=6.89; EPGC=4.91; ROGC=7.78 and ROEP=6.20. Vitamin C and trolox showed 30.11 and 30.10, respectively. Dimers showed greater activity in scavenging free radicals, possibly related to the number of hydroxyls. However, compounds without a hydroxyl at position 5 of the A-ring (5-deoxy-proanthocyanidins) did not change the antioxidant activity of the DPPH free radical, as evaluated here for the first time. Among the monomers, there appeared to be a direct relationship in scavenging of free radicals because of the stereochemistry of the compounds. The presence of a methyl radical on the B-ring significantly reduced the scavenging of free radicals of gallocatechin. All compounds showed greater scavenging of radicals than vitamin C and trolox, and these two compounds showed no significant difference from each other.

O extrato das cascas de Stryphnodendron obovatum submetido à cromatografia em coluna, Sephadex® LH-20, forneceu nove substâncias: ácido gálico (GA), ácido p-hidróxibenzóico (PHB), galocatequina (GC), epigalocatequina (EPG), 4'-O-metilgalocatequina (MGC), epigalocatequina-(4β→8)-epigalocatequina (EPEP), epigalocatequina-(4β→8)-galocatequina (EPGC), robinetinidol-(4α→8)-galocatequina (ROGC) e robinetinidol-(4β→8)-epigalocatequina (ROEP). A capacidade antioxidante in vitro pelos métodos do radical livre DPPH (IC50; μg/mL) e do complexo fosfomolibdênio (CAR) apresentou os seguintes resultados, respectivamente: extrato bruto 4,52 e 0,8242; fração acetato de etila 4,04 e 0,9537 e fração aquosa 5,58 e 0,9275 demonstrando possuírem capacidade antioxidante quando comparados com vitamina C 4,93 e 1,0. Os valores obtidos pelo método do radical livre DPPH com as substâncias isoladas foram: IC50 (μM): GA=8,89; PHB=10,12; GC=16,46; EPG=13,20; MGC=21,00; EPEP=6,89; EPGC=4,91; ROGC=7,78 e ROEP=6,20. Vitamina C e trolox mostraram valores de 30,11 e 30,10, respectivamente. Os dímeros mostraram maior atividade no sequestro de radicais livres, possivelmente relacionada com o número de hidroxilas. No entanto, substâncias com ausência de hidroxila na posição 5 do anel A (5-desoxi-proantocianidinas) não alteraram o poder antioxidante frente ao radical livre DPPH, tendo sido avaliadas aqui pela primeira vez. Entre os monômeros, parece haver uma relação direta com a estereoquímica. A presença de um radical metila no anel B reduziu significativamente a atividade da galocatequina. Todas as substâncias isoladas mostraram maior atividade do que a vitamina C e trolox, e estes não mostraram diferença significativa entre si.