Effect of Tamarindus indica L. Leaf Extract and Citrus sinensis L. Osbeck Peel Extract on Meloidogyne spp (Root Knot Nematodes) in Jos, Plateau State

Aim: The Aim of this study is to determine the effect of aqueous leaf extract of Tamarindus indica and peel extract Citrus sinensis on Root Knot Nematodes, in vitro. Study Design: The experiment was carried out using a completely randomized design. Place of Study: This study was carried out in the biology laboratory of the Department of Plant Science and Biotechnology, University of Jos, Nigeria. Methods: Extraction of phytochemicals from Tamarindus indica leaf and Citrus sinensis peels was carried out using an aqueous solvent. Tamarindus indica leaves and Citrus sinensis peels were collected within the province of Jos, and were dried at constant room temperature to a constant weight after which pulverized, sieved and soaked in water for 24 hours. The extraction was done using the cold maceration method. Eighty (80) second-stage juveniles of Meloidogyne spp. were exposed to the extracts at different concentrations (100, 80, 60, 40, and 20mg/ml) for 6, 12, 18, 24. 30, 32 and 48 hours and, each treatment was replicated 5 times. The average number of dead nematodes per concentration were determined and recorded as mean ± SEM with statistical value at P < 0.05 considered significant. Results: Results of phytochemical analysis showed that Alkaloids, Flavonoids, Tannins, Steroids, Phenols, and Cardioglycosides were present in T. indica leaf extracts while C. sinensis peels contains Flavonoids, Alkaloids, Saponins, Tannins, Phenols, and Resins. These phytochemicals present are suspected to be responsible for the nematicidal activities of the plants. Nematode mortality in the two treatment groups (T. indica and C. sinensis extracts) was highest at 100mg/ml concentration (48 hours of exposure); It was followed by 80mg/ml in decreasing order while the least mortality rate was recorded at 20mg/ml. Hence, as the concentration of the extracts were increased with increase in time, high nematode mortality was observed. Percentage mortality of treated nematodes was statistically significant (P < 0.05) for all concentrations of the extracts at the different time intervals when compared to the control. Statistical analysis showed that T. indica had significantly higher effect on Root-Knot nematodes mortality than C. sinensis at varying concentrations and time of exposure. Benefits of Results: Synthetic pesticides are the principal means used to control nematodes, but the natural products may provide a safe alternative. Hence, the active compounds (phytochemicals) in T. indica L. and C. sinensis L. can be harnessed to be used as biopesticides. Conclusion: The study suggests that extracts of T. indica and C. sinensis can be utilized in the control of root knot nematodes and may have the potential for the production of biopesticides that can be used for the control of nematodes as it displayed nematicidal activities.

Bacterial Culture Filtrates Antinematode Activity of Abomasum Bacterial Culture Filtrates against Haemonchus contortus in Small Ruminants

Haemonchosis is a parasitic disease of small ruminants that adversely affects livestock production. Haemonchus contortus is one of the most prevalent nematode parasites that infect the abomasum of small ruminants. This parasite reduces milk production, overall growth and sometimes causes the death of the infected animals. The evaluation of the biocontrol potential of some abomasum bacterial isolates against H. contortus is investigated in this study. Out of which, three isolates—Comamonas testosteroni, Comamonas jiangduensis, Pseudomonas weihenstephanesis—show significant effect against the nematode L3, adult, and egg hatch inhibition assays. Various concentrations of metabolites from these bacteria are prepared and applied in different treatments compared with control. In the case of adult mortality assay, 50% metabolites of C. testosteroni and P. weihenstephanesis show 46% adult mortality, whereas C. jiangduensis shows 40% mortality. It is observed that decreasing the concentration of bacterial metabolite, lowers nematode mortality. The minimum nematode mortality rate is recorded at the lowest filtrates concentration of all the bacterial isolates. The same trend is observed in egg hatch inhibition assay, where the higher concentration of bacterial culture filtrates shows 100% inhibition of H. contortus egg. It is concluded that the effect of bacterial culture filtrates against H. contortus is dose-dependent for their activity against nematode L3, adult, and inhibition of egg hatchment.

EFFECT OF HYDROSTATIC PRESSURE ON THE VIABILITY OF EPN NATIVE ITALIAN STRAINS STEINERNEMA FELTIAEAND HETERORHABDITIS BACTERIOPHORA

The use of entomopathogenic nematodes (EPNs)as a non-chemical treatment system is one of the most mo-dern techniques promoted by the 2008/128/EC standards for the biological control of plant pathogens. The distri-bution ofEPNscould be carried out with the same sprayers used for the distribution of pesticides. This research is focusedon the effects of the mechanical stress due to hydraulic compression on EPNsItalianstrains,Stei-nernemafeltiaeand Heterorhabditisbacteriophora, to assess their residual strength and vitality under laboratory tests. Liquid suspensions of nematodes were subjected to 8 different levels of hydrostatic pressure from 5 to 40 bar. The hydraulic compression test was conducted through a hydrostatic compression system specifically de-signed for the experimental test. Although correlation curves show an increment of14%in nematode mortality, the statistical analysisdid not show any significant effect of static pressure on the vitality and infectivity of EPNs, that makes them suitable for field application with sprayers. These preliminary positive results on the effects of compression on the viability of EPNsare preparatory to subsequent field tests under high pressure

Nematicidal Activity of Herbal Crude Extract Against Meloidogyne Incognita in Musa Acuminate (Red Banana)

The current study was assumed to identify the novel activities from different plant species for the control of nematodes. To resolve the egg hatchability and Nematode mortality rates of root knot Nematodes. Root knot Nematodes are most abundant and destructive Nematode around all the tropics and sub tropic regions of the world. These root knot Nematodes isolated from the Musa acuminata were treated with extracts of five plant leaves namely Azadirachta indica (neem), Arthemisia pallens (marikolunthu), Ocimum tenuiflorum (tulasi), Hibiscus rosasinensis (Sembaruthi), Ficus hispida (fig) with methanol were evaluated against root knot Nematodes Meloidogyna incognita under laboratory conditions. The Nematicidal activity of Methnolic and leaves of five plant species extract, decreases the Nematodes viability as the concentration of the extracts increases. Hence they examine the Nematicidal capability of the plant species against Meloidogyna incognita. This article reports a reduction in egg hatching and increased Nematicidal activity.

Biological Control Associated With Plant Nutrition for Meloidogyne javanica and Pratylenchus brachyurus Management in Soybean

Meloidogyne javanica and Pratylenchus brachyurus stand out among the main nematodes in soybean crops. Research on integrated management are often conducted, due to the low efficiency of the main control methods when they are applied alone. Thus, the aim of the present study was to assess the potential of biological control and plant nutrition products to control these nematodes in soybean. The effect of each product alone on nematode hatching and mortality was also assessed. A greenhouse experiment was also carried out, evaluating five doses of the product for biological control based on Bacillus and Trichoderma, with and without the presence of the product for nutrition, inoculated with 2000 eggs and juveniles for the gall nematode or 1000 specimens for the nematode lesions. After 30 days of multiplication, the aerial part was removed and the soil was revolved to receive the new sowing of the soybean with the respective treatments mentioned above. After 60 days, the experiments were evaluated for nematological parameters. Both products reduced hatching and increased nematode mortality. Treatments with biological control were efficient in reducing M. javanica and P. brachyurus, mainly when applied at doses close to 5 and 8 kg ha-1, respectively. The nutrition product negatively influences the biological control.

CONTROLE BIOLÓGICO DE Bursaphelenchus cocophilus POR RIZOBACTÉRIAS E Trichoderma spp.

O objetivo do trabalho foi avaliar o potencial de rizobactérias e Trichoderma spp. no controle biológico de Bursaphelenchus cocophilus. Foram avaliados 19 isolados de Pseudomonas spp. (P), 29 de Bacillus spp. (B) e 27 de Trichoderma spp. (T), obtidos de plantio comercial. Os isolados foram caracterizados quanto a produção de compostos bioquímicos. Foi avaliada a taxa de mortalidade de B. cocophilus in vitro e in vivo, neste último realizou-se também a quantificação das enzimas relacionadas à patogênese. Os isolados B14 e P41 proporcionaram 69% e 56% de taxa de mortalidade de nematoides, respectivamente e foram tanto proteolíticos como solubilizadores de fosfato. Os isolados T41 e T54 apresentaram 96% de mortalidade de B. cocophilus in vitro e foram produtores de quitinases, protease e sideróforos. No experimento in vivo, os isolados de rizobactérias B14, B23 e P23 promoveram maior taxa de mortalidade do B. cocophilus do que os demais tratamentos. As mudas de coqueiro inoculadas com o B. cocophilus apresentaram aumento nas atividades das enzimas peroxidases, quitinase e β-1,3-glucanases, quando comparadas com a testemunha (mudas não inoculadas). Os isolados B41, P14, T41 e T54 apresentaram-se como potenciais agentes de controle biológico do B. cocophilus.Palavras-chave: anel vermelho, antagonismo, nematoid. BIOLOGIC CONTROL OF Bursaphelenchus cocophilus WITH RHIZOBACTERIA AND Trichoderma isolates ABSTRACT:The purpose of this study was to evaluate the potential of rhizobacteria Pseudomonas spp. and Bacillus spp. and Trichoderma spp. isolates on the biological control of the nematode Bursaphelenchus cocophilus. The potential biological control isolates (48 rhizobacteria and 27 Trichoderma spp.) were characterized biochemically. The biological control experiments were carried out "in vitro" and "in vivo", and in the later one, the mortality rate of the nematodes and the pathogenesis related enzymes were evaluated. Rhizobacterial isolates B14 and P41 provided a high nematode mortality rate (69% and 56%, respectively) and both were proteolytic and phosphate solubilizers. In the "in vivo" experiment, rhizobacterial isolates B14, B23 and P23 tested the highest mortality rates of B. cocophilus than the isolates of Trichoderma spp. and of the control as coconut saplings challenged with B. cocophilus showed indices higher in enzyme activities, peroxidases, chitinase and β-1,3-glucanases when compared to witness (seedling not inoculated). The isolates B41, P14, T41 and T54 were presented as potential biological control agents of B. cocophilus.Keywords: red ring, antagonism, nematode. DOI:

The Effect of Bacteria Colony Pseudomonas fluorescens (UB_Pf1) and Bacillus subtilis (UB_Bs1) on the Mortality of Pratylenchus coffeae (Tylenchida: Pratylenchidae)

<p>Parasitic Root-Lession nematode of <em>Pratylenchus coffeae</em> can reduce the Indonesian coffee plants productivity. Several studies reported that <em>Pseudomonas fluorescens </em>and <em>Bacillus subtilis </em>endophytic bacteria were antagonistic bacteria to nematode. The objective of this research was to reveal the effectiveness of bacterial colonies density of <em>P. fluorescens </em>(UB_Pf1), <em>B.subtilis </em>(UB BS1), and a combination of both bacteria on nematode mortality using median lethal concentration (LC₅₀) and median lethal time 50 (LT₅₀). The densities of bacteria used in this study were 10⁷, 10⁹, 10¹¹and 10¹³cfu/ml. 35 testing nematodes were used and the mortality was counted at 6, 12, 24, 36, and 48 hours after treatments. The results showed that LC₅₀ values of <em>P. fluorescens </em>was (UB_Pf1) was 4,3x10⁸cfu/ml, LC₅₀<em>B. subtilis </em>(UB_Bs1) was 1,9x10⁹cfu/ ml, and LC₅₀combination of both bacteria was, 8x10⁷cfu/ml. It implies that the application of the combination of both bacteria are more pathogenic than single bacterial treatment. The results also showed that the highest LT₅₀value was 13.21 hours combination of bacterial colonies with a density of 10¹³cfu/ml and the lowest LT₅₀value was 52.00 hours on <em>P. fluorescens </em>(UB_Pf1) treatment with colonies density of 10⁷cfu/ml.</p><p><strong>How to Cite</strong></p><p>Purwaningtyas, P. M., Rahardjo, B. T., &amp; Tarno, H. (2016). The Effect of Bacteria Colony <em>Pseudomonas fluorescens</em> (UB_Pf1) and <em>Bacillus subtilis</em> (UB_Bs1) on the Mortality of <em>Pratylenchus coffeae</em> (Tylenchida: Pratylenchidae). <em>Biosaintifika: Journal of Biology &amp; Biology Education</em>, 8(3), 286-293.</p><p> </p>

Microbiota from Meloidogyne exigua egg masses and evidence for the effect of volatiles on infective juvenile survival

Egg masses ofMeloidogyne exiguafrom coffee plants are subjected to a range of microbial populations and these resident soil organisms in the egg masses may affect nematode communities. The incidence of fungi and bacteria inM. exiguaegg masses was investigated and the toxic effect of their volatile organic compounds (VOCs) was tested on second-stage juveniles (J2). Bacteria and fungi were isolated from egg masses on coffee root and identified to species. The nematicidal activity of VOCs from bacterial and fungal strains was tested againstM. exiguainin vitroexperiments. Several bacterial and fungal strains were found inM. exiguaegg masses and produced VOCs that were toxic toM. exiguaJ2. Bacterial strains induced more nematode mortality compared with fungal strains. The continued colonisation of fungi and bacteria inM. exiguaegg masses occurred during the year and their VOCs reduced the viable inocula ofM. exiguaand should be explored as biocontrol agents.

UJI KESESUAIAN EMPAT ISOLAT TRICHODERMA SPP. DAN DAYA HAMBAT IN VITRO TERHADAP BEBERAPA PATOGEN TANAMAN

Compatibility test of four Trichoderma spp. Isolates and in vitro inhibition ability on several plant pathogens. In vitro descriptive compatibility research was carried out to know the compatibility among Trichoderma spp. isolates and their inhibition ability toward several plant pathogens. Four Trichoderma spp. isolates used were ginger, shallot, banana, and pineapple isolates; while the pathogens used were pathogenic fungi (Fusarium, Colletotrichum, Phytophthora, and Sclerotium), bacteria (Ralstonia), and nematode (Meloidogyne dan Globodera). Observation was done toward inhibition zone between Trichoderma spp. isolates, colony radial growth, mycelial dry weight, and nematode mortality. Result of the research indicated that the four Trichoderma spp. isolates were compatible and no growth inhibition was observed. The inhibition ability of all Trichoderma spp. isolates varied and the ginger isolate had the highest inhibition ability and mortality on all fungal species and the nematodes, while on the pathogenic bacteria there was no inhibition.

Nematicidal effect of volatile organic compounds (VOCs) on the plant-parasitic nematode Meloidogyne javanica

Previous studies have demonstrated that volatile organic compounds (VOCs), produced by the yeast Saccharomyces cerevisiae, were able to inhibit the development of phytopathogenic fungi. In this context, the nematicidal potential of the synthetic mixture of VOCs, constituted of alcohols and esters, was evaluated for the control of the root-knot nematode Meloidogyne javanica, which causes losses to crops of high economic value. The fumigation of substrate containing second-stage juveniles with VOCs exhibited nematicidal effect higher than 30% for the lowest concentration tested (33.3 µL g-1 substrate), whereas at 66.6 and 133.3 µL g-1 substrate, the nematode mortality was 100%. The present results stimulate other studies on VOCs for nematode management.