Control potential of Meloidogyne javanica and Ditylenchus spp. using fluorescent Pseudomonas and Bacillus spp

ABSTRACT Plant Growth Promoting Rhizobacteria (PGPR) have different mechanisms of action in the development of plants, such as growth promotion, production of phytohormones and antibiotic substances and changes in root exudates. These help to control plant diseases. In order to evaluate the potential of microorganisms in the control of Meloidogyne javanica and Ditylenchus spp., five rhizobacteria isolated from rhizosphere of garlic cultivated in the Curitibanos (SC) region were tested. Hatching chambers were set on Petri dishes, in which were added 10 mL of bacterial suspension and 1 mL of M. javanica eggs suspension, at the rate of 4500, on the filter paper of each chamber. The same procedure was performed with 300 juvenile Ditylenchus spp. The experimental design was completely randomized, with four replications. The evaluations were performed every 72 h for nine days. The antagonized population of nematodes was determined in Peters counting chamber, determining the percentage hatching (for M. javanica) and motility (for Ditylenchus spp). Isolates CBSAL02 and CBSAL05 significantly reduced the hatching of M. javanica eggs (74% and 54.77%, respectively) and the motility of Ditylenchus spp. (55.19% and 53.53%, respectively) in vitro. Isolates were identified as belonging to the genera Pseudomonas (CBSAL05) and Bacillus (CBSAL02).

Meloidogyne and Pratylenchus species in sugarcane fields in the state of Alagoas, Brazil / Espécies de Meloidogyne e de Pratylenchus em áreas cultivadas com cana-de-açúcar no estado de Alagoas

ABSTRACT: The objective of this study was to accomplish a survey on populations of Meloidogyne and Pratylenchus species in sugarcane farming areas in the state of Alagoas, Brazil. Twenty samples of soil and roots were processed to extract and quantify nematodes; however, the identification of Meloidogyne species was performed using only 12 samples. Pratylenchus spp. were reported at moderate population levels of 68-1556 specimens 50g-1 of roots and 2-298 specimens 100cm-3 of soil in twenty analyzed samples. For Meloidogyne spp., these values were of 12-487 specimens 50g-1 of roots and 0-140 specimens 100cm-3 of soil. Based on electrophoresis of esterase isozymes, M. incognita was reported to be the most frequent species, followed by M. javanica and M. arenaria. Pratylenchus species identified through morphometrical and morphological characteristics were P. zeae and P. brachyurus , with predominance for the first species. No significant correlation (P≤0.05) were reported between nematode populations and sugarcane cropping systems.

RESUMO: O objetivo deste trabalho foi realizar um levantamento de espécies de Meloidogyne e de Pratylenchus existentes em áreas de cultivo de cana-de-açúcar, no estado de Alagoas. Vinte amostras de solo e raízes foram processadas para a extração e quantificação, sendo a identificação das espécies de Meloidogyne realizada em 12 amostras. Detectou-se a presença de Pratylenchus spp. em níveis populacionais médios de 68-1556 espécimes 50g-1 de raízes e 2-298 100cm-3 de solo, em todas as amostras. Para Meloidogyne spp., esses valores foram de 12-487 50g-1 de raízes e 0-140 100cm-3 de solo. Com base na eletroforese da isoenzima esterase, M. incognita foi a espécie mais frequente, seguida por M. javanica e M. arenaria . As espécies de Pratylenchus identificadas pelas características morfológicas e morfométricas, foram P. zeae e P. brachyurus , com predomínio da primeira espécie. Não houve correlações significativas (P≤0,05) entre as populações de nematoides e as características dos sistemas de cultivo de cana-de-açúcar.

Leafy Vegetables Free Of Coliforms And Parasites After Washing In Chlorine Treated Water

The production and preparation of vegetables for consumption involve procedures that pose health risks to consumers. In this context the water plays an important role in the quality of minimally processed vegetables. We assessed the hygienic-sanitary quality of leafy vegetables and obtained data of interest to health surveillance agencies and public health. They were tested for the presence of total and faecal coliforms, and also parasitic protozoa and helminths. The vegetable samples were purchased in supermarkets of the Midwest of the State of Santa Catarina, Brazil. We performed bacteriological analyses of wash water samples using chromogenic substrates to obtain the most probable number of the coliform group at 35 °C and Escherichia coli. We used Hoffman's spontaneous sedimentation technique for the parasitological analysis in order to check the occurrence of heavy and light helminths eggs and protozoan cysts. The analyses indicated the presence of coliforms, nematodes and freeliving protozoa, except for the last wash. We concluded that the washing process using running water and no active chlorine was not efficient in reducing the load of microorganisms in the vegetables. Therefore, intensive educational programmes should be implemented by health authorities to encourage hygienic-sanitary practices and risk reduction of food-borne diseases.(AU)

New record of Pelecitus sp. (Nematoda, Onchocercidae) as a parasite of Athene cunicularia (Strigiformes, Strigidae) in southeastern Brazil / Novo registro de Pelecitus sp. (Nematoda, Onchocercidae) como parasita de Athene cunicularia (Strigiformes, Strigidae) no Sudeste do Brasil

The aim of this study was to report the burrowing owl Athene cunicularia as a new host for the filarid nematode Pelecitus sp. in southeastern Brazil for the first time, as well as reporting the occurrence of this nematode species in the body cavity, near the cervical air sac and lung region. This study contributes towards knowledge of parasitism in Brazilian wild birds and an anatomical region of the host as an infection site for Pelecitus sp.

O objetivo do estudo foi relatar, pela primeira vez, a coruja buraqueira Athene cunicularia como um novo hospedeiro para o nematóide filarídeo Pelecitus sp. do Sudeste do Brasil, bem como a ocorrência desta espécie de nematóides na cavidade corporal, próximos à região dos sacos aéreos cervicais e pulmões. Este estudo contribui para o conhecimento do parasitismo em aves silvestres brasileiras e uma região anatômica do hospedeiro como sítio de infecção para Pelecitus sp.

Romanomermis culicivorax (Nematoda:Mermithidae): una alternativa para el control de vectores de malaria en Nicaragua / Romanomermis culicivorax (Nematoda: Mermithidae): an alternative for malaria vector control in Nicaragua

Introducción: la aparición de casos de malaria en zonas donde se creía controlada, como lo es la Región Autónoma Atlántica Norte en Nicaragua, se debe a múltiples factores como el costo de los productos químicos para el control vectorial, zonas de difícil accesibilidad y altos índices de pobreza. Recientemente existe un marcado interés en disminuir la incidencia y prevalencia de la malaria en esta área, mediante la producción y aplicación de alternativas biológicas de control. Objetivos: establecer y estandarizar la cría de Romanomermis culicivorax para la producción masiva en condiciones de laboratorio y su utilización en criaderos naturales para reducir la población de Anopheles albimanus en 3 municipios de la Región Autónoma Atlántica Norte. Métodos: se utilizaron cultivos de una cepa de Romanomermis culicivorax provenientes del Instituto de Medicina Tropical Pedro Kourí de La Habana, Cuba. El establecimiento y la producción de esta cepa se hizo siguiendo la metodología descrita por especialistas cubanos. El trabajo se realizó en 7 criaderos de 5 barrios pertenecientes a los municipios Puerto Cabezas, Waspam y Rosita, desde noviembre de 2009 a marzo de 2011. Resultados: se estableció y estandarizó la cría del nematodo, utilizando la dosis de infestación de 7:1 (7 pre-parasíticos por larva) en condiciones de laboratorio. La dosis de aplicación en los criaderos fue 1 000 pre-parasíticos/m²,demostrando ser adecuada para las condiciones de campo. Se lograron porcentajes de reducción elevados y en menor tiempo en los municipios Waspam y Rosita. Se demostró la permanencia del nematodo en los criaderos. Conclusiones: se establece por primera vez y con éxito la producción masiva de Romanomermis culicivorax en Nicaragua. Los resultados en el terreno demostraron la eficiencia de este nematodo para controlar densidades larvales de Anopheles albimanus, lo cual constituye una alternativa de control vectorial(AU)

Introduction: the emergence of malaria cases in areas where the disease was thought to be controlled, such as the North Atlantic Autonomous Region in Nicaragua, is due to a variety of factors, such as the cost of chemicals for vector control, the inaccessibility of areas and the high rates of poverty. A marked interest has recently arisen in reducing the incidence and prevalence of malaria in the region by developing and implementing biological control alternatives. Objectives: establish and standardize the breeding of Romanomermis culicivorax for mass production under laboratory conditions and for its use in natural breeding sites to reduce the population of Anopheles albimanus in three municipalities of the North Atlantic Autonomous Region. Methods: the study was based on cultures of a Romanomermis culicivorax strain obtained from Pedro Kourí Tropical Medicine Institute in Havana, Cuba. The strain was established and produced following the methodology described by Cuban specialists. The research was conducted at 7 breeding sites from 5 districts in the municipalities of Puerto Cabezas, Waspam and Rosita, from November 2009 to March 2011. Results: breeding of the nematode was established and standardized with a 7:1 infestation dose (7 preparasites per larva) under laboratory conditions. The application dose at breeding sites was 1 000 preparasites/m², which proved to be appropriate for field conditions. High reduction percentages were obtained. These were achieved in a shorter time in the municipalities of Waspam and Rosita. It was found that the nematodes remained at the breeding sites. Conclusions: successful mass production of Romanomermis culicivorax is established for the first time in Nicaragua. Field results show the efficiency of this nematode to control larval densities of Anopheles albimanus, constituting a vector control alternative(AU)

Structural and functional analysis of cathepsin S of Heterodera spp: A promising candidate for its control.

Cysteine proteinases are required for a wide range of physiological processes in all living organisms. In parasitic nematodes, they are particularly crucial for the digestion of host tissues and evasion of host immune responses. Therefore, in general, these are identified as primary targets for the control of parasitic nematodes. Herein, cathepsin S-like cysteine proteinase of Heterodera avenae (Hacp-s) has been cloned and analysed for the first time. The predicted protein is 298 amino acids long and showed significant similarity with cathepsin S of Heterodera glycines (Hgcp-s). The sequence of cathepsin S contains a signal peptide of 30 amino acids which suggests its role in extracellular functions. Multiple sequence alignment revealed the presence of ERFNIN motif and conserved catalytic residues. Three dimensional structure (3D) of Hgcp-s was modelled using homology modelling. In order to illustrate the plausible mode of interaction of cathepsin S (Hgcp-s), docking analysis was performed with E-64 cysteine proteinase inhibitor. Docking studies revealed the hydrogen bonding of E-64 with Gln153, His299 and Gly203 as well as close interaction with catalytic residues Cys159 and Asn320. Expression analysis of Hacp-s using qRT-PCR showed high expression of cathepsin S in pre parasitic J2s and female stages suggesting its significant role in both pre-parasitic and parasitic stages of the nematode life cycle.

COMPORTAMIENTO DE MATERIALES DE GUAYABO (Psidium guajava LINNEO) AL PARASITISMO DEL NEMATODO FORMADOR DE AGALLAS [ Meloidogyne incognita (KOFOID & WHITE) CHITWOOD Y Meloidogyne javanica (TREUB) CHITWOOD] / BEHAVIOR OF GUAVA MATERIALS (Psidiumguajava LINNEO) ON THE ROOT KNOT NEMATODE [ Meloidogyne incognita (KOFOID & WHITE) CHITWOOD AND Meloidogyne javanica (TREUB) CHITWOOD] PARASITISM

Las raíces del guayabo son parasitadas por el nematodo del nudo radical (Meloidogyne spp.), ocasionando pérdidas de la producción entre 30 y 60%. Debido a esto, el principal objetivo de esta investigación fue evaluar en condiciones de almácigo la reacción de cuatro variedades de guayabo al ataque del complejo Meloidogyne incognita y M. javanica. En el vivero Jaibaná Frutales, del municipio de Pereira (Risaralda), plántulas de cuatro variedades: Pera y Común (Psidium guajava), Agrio (Psidium friedrichsthalianum) y Peruano (Psidium cattleianum), fueron sembradas en bolsas de plástico que contenían suelo y cascarilla de arroz en proporción 3:1. Veinte días después de sembradas, a 15 plántulas de cada una de las variedades le fueron inoculadas una mezcla de 1.000, 5.000 y 10.000 huevos y juveniles de Meloidogyne, y los respectivos testigos sin inocular. Las plántulas fueron asignadas bajo un diseño experimental completamente aleatorio. Las variedades Peruano con 5,75g y 8,03g de peso seco de raíces y parte aérea, respectivamente, y Agrio con 10,27g y 9,23g de peso seco de raíces y parte aérea, respectivamente, presentaron el menor índice de nudosidad con 0,1 y 1,1, y reproducción de Meloidogyne spp. con valores entre 75 y 4.575 huevos y juveniles/100g de raíces. Estas variedades fueron estadísticamente diferentes a la variedad Pera con 13,96g y 11,33g de peso seco de raíces y parte aérea, respectivamente, y Común con 11,35g y 13,9g de peso seco de raíces y parte aérea, respectivamente, quienes presentaron mayor índice de nudosidad con 5 y reproducción con 149.775 huevos y 72.950 juveniles/100g de raíces. Las variedades Agrio y Peruano fueron resistentes e inmunes, respectivamente, al ataque de Meloidogyne incognita y M. javanica, permitiendo ser utilizadas como portainjertos o patrones; mientras que las variedades Pera y Común fueron altamente susceptibles.

Guava roots are parasitized by the root knot nematode (Meloidogyne spp.), which causes production losses between 30 and 60%. Due to this, the main goal of this research was to evaluate the reaction of four varieties of guava under nursery conditions to the attack of Meloidogyne incognita and M. javanica complex. In the Jaibaná Frutales nursery, in the municipality of Pereira (Risaralda), seedlings of four varieties of guava: Guava pear and Common guava (Psidiumguajava), Cas guava (Psidiumfriedrichsthalianum), and Peruvian guava (Psidiumcattleianum), were planted in plastic bags containing soil and rice hull in a 3:1 proportion. Twenty days after planted, 15 seedlings of each variety were inoculated with a mixture of 1,000, 5,000 and 10,000 eggs and Meloidogyne spp juveniles, and their respective controls without inoculation. Seedlings were assigned under a completely randomized design. The Peruvian guava variety with 5.75g and 8.03g roots and aerial parts dry weight respectively, and Cas guava variety with 10.27g and 9.23g roots and aerial parts dry weight, respectively, had the lowest root gall index with 0.1 and 1.1, and Meloidogyne spp. reproduction with values between 75 and 4.575 eggs and juveniles/100g of roots. These varieties were statistically different from the guava Pear variety with 13,96g y 11,33g roots and aerial parts dry weight respectively, and Common guava with 11,35g y 13,9g roots and aerial parts dry weight respectively, which had the highest root gall index with 5 and reproduction with 149.775 eggs and 72.950 juveniles/100g of roots. In conclusion, the Cas and Peruvian varieties were resistant and immune respectively, to Meloidogyne incognita and M. javanica, allowing them to be used as rootstocks or patronages, while guava Pear and Common guava were highly susceptible to both species.

Conjoint effect of oil-seed cakes and Pseudomonas fluorescens on the growth of chickpea in relation to the management of plant-parasitic nematodes

Soil application of organics has been explored as an alternative means of organic management of plant-parasitic nematodes. Efficiency of different oil-seed cakes of neem (Azadirachta indica), castor (Ricinus communis), groundnut (Arachis hypogaea), linseed (Linum usitatissimum), sunflower (Helianthus annuus) and soybean (Glycine max) were evaluated in field conditions with association of Pseudomonas fluorescens in relation to growth parameters of chickpea and population of plant-parasitic nematodes. Their efficacious nature was highly effective in reducing the population of these dominant soil nematodes. Significant improvement was observed in plant-growth parameters such as plant weight, percent pollen fertility, pod numbers, root-nodulation and chlorophyll content of chickpea, seemed to be due to reduction in disease incidence and might be due to growth promoting substances secreted by P. fluorescens. The multiplication rate of nematodes was less in the presence of P. fluorescens as compared to its absence. Most effective combination of P. fluorescens was observed with neem cake.