Abstract

BackgroundBiological control using entomopathogenic nematodes (EPN) has demonstrated good potential to contribute to the integral control of mosquito larvae, which as adults are vectors of diseases such as Dengue fever, Zika and Chikungunya. However, until now there are no records of the presence of EPN or their killing capacity in Yucatán state, southern México. The objectives of the current study were: (1) to report the entomopathogenic nematodes present in Yucatán soils and (2) to determine the killing capacity of the most frequent and abundant EPN against Aedes aegypti mosquito larvae and the microbial community developed by Ae. Aegypti exposed to this EPN.MethodsThe nematodes were collected by the insect trap technique using the great wax moth Galleria mellonella. Internal transcribed spacer (ITS), 28S gene of ribosomal DNA and phylogenetic analyses were performed to identify the EPN. For the bioassay, four concentrations of the most frequent and abundant EPN were tested: 1,260:1 infective juveniles (IJs) per mosquito larvae, 2,520 IJs:1, 3,780 IJs:1 and 5,040 IJs:1. High-throughput sequencing of the 16S rRNA gene was used to identify bacterial amplicon sequences in the mosquito larvae infected with EPN.ResultsSix isolates of Heterorhabditis were recovered from 144 soil samples. Heterorhabditis indica (four isolates) was the most frequent and abundant EPN, followed by Heterorhabditis n. sp. (two isolates). Both nematodes are reported for the first time for Yucatán state, Mexico. The concentration of 2,520 IJs:1 produced 80% of mosquito larvae mortality in 48 h. Representative members of Photorhabdus genus were numerically dominant (74%) in mosquito larvae infected by H. indica. It is most likely that these bacteria produce secondary toxic metabolites that enhance the mortality of these mosquito larvae.

Highlights

  • Biological control using entomopathogenic nematodes (EPN) has demonstrated good potential to contribute to the integral control of pests of agricultural and medical importance (Chitra, Sujatha & Alagarmalai, 2017)

  • Molecular identification and phylogenetic analysis of Heterorhabditis The comparison of nucleotide sequences with the sequences published in GenBank through BLAST allowed us to molecularly identify our EPN from Yucatán state, where some of our specimens had a high percentage of similarity with H. indica (99.73–100% in Internal transcribed spacer (ITS) and 99.89–100% in 28S)

  • The phylogenetic tree obtained with the ITS data set showed that the new sequences of this study were grouped into two clades, one corresponding to H. indica and another one that belongs to an unidentified species: Heterorhabditis n. sp. (Fig. 2)

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Summary

Introduction

Biological control using entomopathogenic nematodes (EPN) has demonstrated good potential to contribute to the integral control of pests of agricultural and medical importance (Chitra, Sujatha & Alagarmalai, 2017). In the case of nematodes of the families Steinernematidae and Heterorhabditidae, their killing capacity is based on their symbiotic relationship with bacteria of the genera Xenorhabdus and Photorhabdus, as well as on their capacity to infect insect hosts. The bacterial community produces secondary metabolites with cytotoxic, antimicrobial, antiparasitic and insecticide activity, causing septicaemia in a period of 24 to 48 h after infection (Bode, 2009). These nematodes feed on the host tissues and reproduce for two or three generations, depending on the availability of food resources and the size of the host (San-Blas, Gowen & Pembroke, 2008). It is most likely that these bacteria produce secondary toxic metabolites that enhance the mortality of these mosquito larvae

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