Antibiotic resistant Vibrio harveyi isolated from shrimp pond and anti-Vibrio activity of Combretum quadrangulare Kurz. and Mimosa pudica extracts

Aims@#The occurrence of bacterial disease in shrimp ponds is a major problem faced in shrimp farming. Thus, the aims of this study were to isolate and evaluate antibiotic resistant profile of Vibrio harveyi strain isolated from shrimp pond water, as well as to study the potential anti-Vibrio activity of Combretum quadrangulare Kurz. (CQ) and Mimosa pudica (MP) leaves extracts.@*Methodology and results@#Vibrio harveyi WSC103 was isolated from water in white shrimp (Litopenaeus vannamei) culture pond and identified using 16S rRNA gene sequencing analysis. This strain showed characteristics of multidrug-resistant (7 antibiotics). It had become more sensitive to antibiotics (9 out of 10 antibiotics) after plasmid curing. It is showed CQ and MP leaves extracts contain potent bioactive compounds (tannins, flavonoids, steroids, cardiac glycosides and alkaloids) against V. harveyi WSC103. The aqueous, 95% ethanolic and 75% acetone extracts of CQ (MIC value of 3.13-12.50 mg/mL) and MP (MIC value of 3.13-25.00 mg/mL) leaves revealed strong vibriostatic activity, but aqueous and 95% ethanolic extracts in both plants showed vibriocidal activity. The 95% ethanolic extract of both CQ and MP leaves displayed the excellent vibriocidal property with MBC value of 100 mg/mL with zone of inhibition at 11.44 ± 1.01 and 11.78 ± 1.01 mm by agar disc diffusion.@*Conclusion, significance and impact of study@#The isolated Vibrio harveyi WSC103 was successfully characterized as a novel multidrug-resistant strain. The ethanolic C. quadrangulare Kurz. and M. pudica extracts exhibited prominent vibriostatic and vibriocidal capacities. These finding is proven that C. quadrangulare Kurz. and M. pudica extracts would be an alternative anti-Vibrio agent for aquaculture infectious treatment.

Anthelmintic effect of Cassia fistula and Combretum leprosum protein fractions against goat gastrointestinal nematodes / Efeito antihelmíntico das frações proteicas de Cassia fistula e Combretum leprosum contra nematodeos gastrintestinais de caprinos

Abstract In this study, we evaluated the ovicidal and larvicidal activity of protein preparations obtained from Cassia fistula L. and Combretum leprosum Mart. leaves on the gastrointestinal parasites of goats. Protein preparations were obtained after the extraction of C. fistula L. and C. leprosum Mart. leaves, followed by protein fractionation (with ammonium sulfate saturation percentages of 30%, 30%-60%, and 60%-90%) and dialysis, which resulted in protein fractions (called F1, F2, and F3, respectively). The fractions were evaluated by egg hatching (the eggs were recovered in stool samples from naturally infected goats) and larval development tests. The results reveled that the inhibition of hatching of eggs caused by the protein fractions of C. fistula (38%) were similar to that of the control drug, thiabendazole. In addition, the fractions of C. fistula caused significant inhibition (61-69%) of larval development also. However, C. leprosum did not reveal significant inhibition of egg hatching and larval development. We conclude that C. fistula L. showed better ovicidal and larvicidal activity against endoparasites.

Resumo Neste estudo, foram avaliadas as atividades ovicida e larvicida de preparações proteicas de Cassia fistula L. e Combretum leprosum Mart. em parasitas gastrointestinais de caprinos. As preparações proteicas foram obtidas por extração das folhas de C. fistula L. e C. leprosum Mart. seguido pelo fracionamento proteico (com porcentagens de saturação de sulfato de amônio de 30%, 30-60%, 60-90%) e diálise, resultando nas frações proteicas (intituladas F1, F2 e F3, respectivamente). As frações foram avaliadas nos testes de eclosão de ovos (os ovos foram recuperados em amostras de fezes de cabras naturalmente infectadas) e de desenvolvimento larvar. Os resultados revelaram que a inibição da eclosão de ovos causada pelas frações proteicas de C. fistula (38%) foi semelhante à do fármaco controle, o tiabendazol. Além disso, as frações de C. fistula também causaram inibição significativa (61-69%) do desenvolvimento larvar. No entanto, C. leprosum não revelou inibição significativa na eclosão dos ovos e no desenvolvimento larvar. Concluiu-se que C. fistula L. mostrou uma melhor atividade ovicida e larvicida contra endoparasitas.

Individual variability in morphometry and emergence of seeds of Combretum leprosum / Variabilidade individual na morfometria e emergência de Combretum leprosum

Combretum leprosum Mart. is a medicinal species of the Brazilian savanna with good potential potential for use in the regeneration of degraded areas, mainly because it is resistant to burning. This study aimed to evaluate the fruit and seed morphometry, emergence and early plant development of C. leprosum. The seeds of C. leprosum were extracted from mature fruits from five different mother plants in Mossoró-RN. The fruit and seeds lots varied in length, diameter and mass and mother plant 1 had the lowest seed diameters and mass and also the lower averages for all evaluated characteristics of the seedlings. Seeds of mother plant 2, 3 and 5 had a short period of emergence. There was variation in emergence rates among the individuals. Combretum leprosum individuals has variability in emergence and biometrics of seeds, and these characteristics influence the quality of seedlings.

Combretum leprosum Mart. é uma espécie medicinal do cerrado brasileiro com uso potencial na regeneração de áreas degradadas, principalmente por ser resistente à queimada. O objetivo deste trabalho foi avaliar a morfometria de frutos e sementes, emergência e desenvolvimento inicial de C. leprosum. As sementes de C. leprosum foram extraídas de frutos maduros de cinco plantas-mãe diferentes em Mossoró-RN. Os lotes de frutos e sementes variaram em comprimento, diâmetro e massa e a planta-mãe 1 apresentou os menores diâmetros e massa de sementes e também as menores médias para todas as características avaliadas das mudas. As sementes da planta-mãe 2, 3 e 5 tiveram um curto período de emergência. Houve variação nas taxas de emergência entre os indivíduos. Os indivíduos de Combretum leprosum tem variabilidade na emergência e biometria de sementes, e essas características influenciam a qualidade das mudas.

In-vitro antibacterial screening of methanol extracts of three combretum species against seven strains of methicillin resistant staphylococcus aureus (MRSA)

Background: Medicinal plants are valued as sources of natural compounds some of which are effective against several infectious diseases. Plants from the genus Combretum have been used traditionally in various African societies to treat variety of medical problems especially infectious diseases.Objective: In this study, we investigated the antibacterial activity of the methanol extracts from the leaves of three Combretum species namely Combretum hispidum, Combretum racemosum and Combretum platypterum against seven strains of methicillin resistant Staphylococcus aureus (MRSA) in vitro.Materials and methods: Methanol extract from Combretum hispidum, Combretum racemosum and Combretum platypterum were subjected to agar diffusion assay and broth micro-dilution test for the determination of antibacterial activity and minimum inhibitory concentration (MIC), all the extracts were tested against eight different strain of MRSA. Imipenem and ciprofloxacin were used as control.Results: Extract from Combretum racemosum leaves showed significant anti-MRSA activity with zone of inhibition as high as 37 mm and MIC values ranging from 0.16 - 1.25 mg/mL on all tested strains of MRSA. Extracts from the two other species did not exhibit any activity on all tested strain.Conclusion: Methanol extract from C. racemosum is highly effective against MRSA and it could be a potential source of newer antimicrobial agent against MRSA infections

Long-lasting endothelium-dependent relaxation of isolated arteries caused by an extract from the bark of Combretum leprosum / Extrato das cascas de Combretum leprosum causa relaxamento dependente de endotélio de longa duração em artérias isoladas

Objective To describe and to characterize the relaxing effect of an extract of the bark of Combretum leprosum on isolated arterial rings from different animals.Methods Rings (3 to 4mm) from rabbit, rat, or porcine arteries rings were suspended in an organ bath (Krebs, 37°C, 95%O2/5%CO2) to record isometric contractions. After the stabilization period (2 to 3 hours) contractions were induced by the addition of phenylephrine (0.1 to 0.3µM) or U46619 (10 to 100nM), and Combretum leprosum extract was added on the plateau of the contractions. Experiments were performed to determine the potency, duration, reversibility, and to get insights on the potential mechanism involved in extract-induced relaxations.Results In all rings tested, Combretumleprosum extract (1.5μg/mL) was able to cause relaxations, which were strictly endothelium-dependent. In rabbit or rat thoracic aorta rings, the relaxations were reversed by vitamin B12a or L-NG-nitroarginine. In porcine right coronary arteries and rabbit abdominal aorta, extract caused both L-NG-nitroarginine-sensitive and L-NG-nitroarginine-resistant relaxations. In rabbit thoracic aorta, the extract was relatively potent (EC50=0.20µg/mL) and caused relaxations; intriguingly the endothelium continued to produce relaxing factors for a long period after removing the extract. The magnitude of extract-induced relaxations was significantly reduced in the absence of extracellular Ca2+; in addition, the TRPs channels blocker ruthenium red (10µM) was able to revert extract-induced relaxations. Phytochemical analyses indicated that the extract was rich in polyphenol-like reacting substances.ConclusionsCombretum leprosum extract contains bioactive compounds capable of promoting Ca2+-dependent stimulation of endothelial cells which results in a prolonged production of relaxing factors.

Objetivo Descrever e caracterizar os relaxamentos induzidos por um extrato das cascas de Combretum leprosum em anéis de artérias de diferentes espécies de animais.Métodos Anéis (3 a 4mm) de artérias de coelho, rato e porco foram montados em cubas para órgão isolado (Krebs, 37°C, 95%O2/5%CO2) para registro das contrações isométricas. Após um período de estabilização (2 a 3 horas), as contrações foram induzidas com fenilefrina (0,1 a 0,3µM) ou U46619 (10 a 100nM); no platô dessas contrações, adicionamos o extrato Combretum leprosum. Diferentes protocolos foram realizados para determinar potência, duração, reversibilidade e mecanismo dos relaxamentos induzidos pelo extrato.Resultados Em todas as preparações testadas, o extrato de Combretum leprosum (1,5µg/mL) provocou relaxamentos dependentes de endotélio. Em aorta torácica de coelho ou rato, os relaxamentos foram revertidos pela vitamina B12a ou L-NG-nitro-arginina. Em anéis de aorta abdominal de coelho e de artérias coronárias de porco, o extrato causou relaxamentos sensíveis e resistentes à L-NG-nitro-arginina. Em aorta torácica de coelho, o extrato foi relativamente muito potente (EC50=0,20μg/mL) e quando causou relaxamentos; intrigantemente o endotélio continuou a produzir fatores relaxantes por um longo período após remoção do extrato. A magnitude dos relaxamentos induzidos pelo extrato foi significativamente reduzida em ausência Ca2+ extracelular; ademais, o vermelho de rutênio (10μM), um bloqueador de canais TRPs, foi capaz de reverter os relaxamentos induzidos pelo extrato. Análises preliminares indicaram que o extrato continha compostos com reatividade química semelhante à polifenóis.Conclusão O extrato de Combretum leprosum contem compostos bioativos capazes de promover estimulação dependente de Ca2+ das células endoteliais a qual resulta numa produção prolongada de fatores relaxantes.

Antinociceptive effect on mice of the hydroalcoholic fraction and (-) epicatechin obtained from Combretum leprosum Mart & Eich

Previous studies on Combretum leprosum, a tree growing in the Northeastern states of Brazil, have shown antinociceptive effects of the ethanol extract of its leaves and bark, but studies examining its constituents are rare. The objective of this study was to evaluate the antinociceptive effect of the hydroalcoholic fraction (HF) of one of its constituents, the flavonoid (-) epicatechin (EPI), administered orally to mice (20-30 g) in models of chemical nociception, and the possible mechanisms involved. Different doses of HF (62.5 to 500 mg/kg) and EPI (12.5 to 50 mg/kg) were evaluated in models of abdominal writhing, glutamate, capsaicin, and formalin in animals pretreated with different antagonists: naloxone, ondansetron, yohimbine, ketanserin, pindolol, atropine, and caffeine in the abdominal writhing test. To determine the role of nitric oxide, the animals were pretreated with L-arginine (600 mg/kg, ip) in the glutamate test. The HF was effective (P < 0.05) in all protocols at different doses and EPI was effective in the abdominal writhing, capsaicin and glutamate tests (P < 0.05) at doses of 25 and 50 mg/kg. However, in the formalin test it was only effective in the second phase at a dose of 25 mg/kg. The antinociceptive effect of HF was inhibited when HF was associated with yohimbine (0.15 mg/kg), ketanserine (0.03 mg/kg), and L-arginine (600 mg/kg), but not with the other antagonists. HF and EPI were effective in models of chemical nociception, with the suggested participation of the adrenergic, serotonergic and nitrergic systems in the antinociceptive effect of HF.

Screening of Twenty-four South African Combretum and six Terminalia Species (Combretaceae) for Antioxidant Activities

The dried leaves of Combretum and Terminalia species (Combretaceae)were extracted with acetone; hexane; dichloromethane and methanol. Thin layer chromatography (TLC) plates were developed under saturated conditions and sprayed with 0.22;2-diphenyl-1-picryl hydrazyl (DPPH) in methanol for antioxidant screening. Visualization of separated bands exhibiting antioxidant activities enabled the localization and the subsequent identification of the potential active compounds. The acetone and methanol extracts displayed the presence of antioxidant activity after spraying the chromatogram with DPPH. Hexane and dichloromethane extracts did not have any antioxidant activity. C. hereroense had the highest number of active compounds; followed by C. collinum ssp. taborense; which were 16 and 10; respectively. Acetone extracts of all tested Combretum species had 53 active bands and methanol had 55. All Terminalia species extracted with acetone and methanol had antioxidant activity. T. gazensis and T. mollis methanol extracts had 11 and 14 active compounds respectively in one of the solvent systems used. The qualitative DPPH assay on TLC was successfully used in this study to systematically assess the total antioxidant activity of the Combretum and Terminalia species extracts

Screening of Twenty-Four South African Combretum and Six Terminalia Species (Combretaceae) for Antioxidant Activities

The dried leaves of Combretum and Terminalia species (Combretaceae) were extracted with acetone; hexane; dichloromethane and methanol. Thin layer chromatography (TLC) plates were developed under saturated conditions and sprayed with 0.22;2-diphenyl-1-picryl hydrazyl (DPPH) in methanol for antioxidant screening. Visualization of separated bands exhibiting antioxidant activities enabled the localization and the subsequent identification of the potential active compounds. The acetone and methanol extracts displayed the presence of antioxidant activity after spraying the chromatogram with DPPH. Hexane and dichloromethane extracts did not have any antioxidant activity. C. hereroense had the highest number of active compounds; followed by C. collinum ssp. taborense; which were 16 and 10; respectively. Acetone extracts of all tested Combretum species had 53 active bands and methanol had 55. All Terminalia species extracted with acetone and methanol had antioxidant activity. T. gazensis and T. mollis methanol extracts had 11 and 14 active compounds respectively in one of the solvent systems used. The qualitative DPPH assay on TLC was successfully used in this study to systematically assess the total antioxidant activity of the Combretum and Terminalia species extracts