Isolation and Characterization of Photorhabdus Spp. (Enterobacteriaceae) Isolated From Heterorhabditis Bacteriophora in Kashere, Nigeria

Photorhabdus bacteria are symbiotically associated with the infective juveniles of entomopathogenic nematodes of the genus Heterorhabditis. These bacteria after infecting a susceptible host, produces a host of lethal toxins that killthe insect hosts within 24 to 72 hours. Consequently they have emerged and are trending as excellent biological control agents against insect pests of agricultural crops. Therefore, this study aim to isolate, identify and test the pathogenicity of Photorhabdus bacteria isolated from Heterorhabditis bacteriophora, entomopathogenic nematode from Kashere, Gombe State, Nigeria. To this end phenotypic and biochemical tests were conducted. The tests conducted showed that the isolate exhibited characteristics similar to those of Photorhabdus bacteria that have been identified. Last instar larvae of G. mellonella exposed to different concentrations of the bacterial isolate showed some level of susceptibility of the larvae to the bacterial isolate confirming its biological control potential.

Bio-efficacy of entomopathogenic nematodes, Steinernema feltiae and Heterorhabditis bacteriophora against the Cabbage butterfly (Pieris brassicae [L.]) under laboratory conditions

Background Entomopathogenic nematodes (EPNs) have the potential to supersede larvicidal activity for the management of various insect pests. Result Lab experiments were conducted to test the pathogenicity of 2 EPNs local species; Steinernema feltiae and Heterorhabditis bacteriophora at different (IJs/cm2) concentrations against the cabbage butterfly, Pieris brassicae (L.). The native isolate was obtained from soil samples, collected from Rajgarh, Hamachi Pradesh, India. Petri dish bioassay used the EPNs species (S. feltiae HR1 and H. bacteriophora HR2) at the concentrations (0, 10, 20, 40, 80, 160 IJs/cm2). Based on the pathogenicity of the strains, only 2 isolates effectively showed larvicidal activity. The highest (%) (72.08 and 67.42%), at the 2nd instar larval mortality was recorded in the treatments with H. bacteriophora and S. feltiae at160 IJs/cm2, respectively. At the 4th instar larvae, respective larval mortality (85.38, 69.50%) was recorded in treatment with H. bacteriophora, and S. feltiae, respectively, at160 IJs/cm2. In case of pupae, the mortality rates were (62.12, 58.58%) for H. bacteriophora and S. feltiae, respectively, at 160 IJs/cm2; (74 and 12%) for both the tested EPNs, respectively, at 80 IJs/cm2. Percent of P. brassicae larval mortality treated with the tested EPN isolates was significantly higher than the untreated control. Results revealed that the percent of larval mortality significantly increased with the increase in time periods, being maximum at 72 h. S. feltiae and H. bacteriophora, strains showed potent larvicidal activity at low concentration even at 48 and 72 h of exposure. Conclusion This study revealed that the local strains of EPNs (S. feltiae HR1 and H. bacteriophora HR2) were found as a biocontrol agent against P. brassicae.

Factors affecting the fitness of an entomopathogenic nematode (Heterorhabditis bacteriophora SUP strain) for use in biocontrol in a horticultural area of Argentina

Summary The optimum conditions for storage, infectivity and multiplication of the nematode Heterorhabditis bacteriophora SUP strain, isolated from horticultural soils in Argentina, were determined for later use in biocontrol. This strain was able to kill Tenebrio molitor larvae (over 90%) at 16 and 25°C, after 7 and 4 days, respectively, with mortality being significantly higher at 25°C. No infectivity was reported at 4 and 11°C. Higher concentrations (over 150 infective juveniles (IJ) insect−1) produced higher percentages of mortality. However, increasing inoculum density reduced offspring production and increased emergence time. Storage density between 2500 and 7500 IJ ml−1 had no effect on survival of IJ. Survival of IJ maintained at 4 and 16°C was around 80% after 4 weeks storage and 30% after 32 weeks. However, at 25°C, a 60% IJ survival was observed at 4 weeks, decreasing to around 10% after 16 weeks and with no survival at 24 weeks. An overall significant decrease in virulence was observed after 90 and 120 days of storage but this was more pronounced at a storage temperature of 4°C than at 16 and 25°C. IJ stored at 4°C had reduced virulence after 90 and 120 days, but not after 60 days. This study provides useful information about some fundamental environmental conditions influencing important nematode traits of the species H. bacteriophora.

Evaluation of two native entomopathogenic nematodes against Odontotermes obesus (Rambur) (Isoptera: Termitidae) and Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae)

Background Entomopathogenic nematodes (EPNs) are one of the widely studied biological control agents. The present study was conducted to evaluate the efficacy of two EPNs species, Heterorhabditis bacteriophora (Poinar) (Rhabditida: Heterorhabditidae) and Steinernema aciari (Qui, Yan, Zhou, Nguyen and Pang) (Rhabditida: Steinernematidae), isolated locally from soils of Majuli river island, Assam, India against two important subterrenean pests; Odontotermes obesus (Rambur) (Isoptera: Termitidae) and Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae) under laboratory conditions. Results In case of O. obesus, mortality percent was recorded by H. bacteriophora after 72 h. at 300 IJs/termite and by S. aciari at 250 and 300 IJs/termite after 96 h. The lowest LD50 and LT50 values obtained for H. bacteriophora were 13.054 IJs/termite and 26.639 h., respectively, while those of S. aciari were 42.040 IJs/termite and 31.761 h., respectively. With respect to A. ipsilon, H. bacteriophora registered a highest mortality rate at 300 IJs/larvae after 144 h. S. aciari showed 100 percent mortality at 300 IJs/larva after 168 h. The lowest values of LD50 and LT50 for H. bacteriophora were 35.711 IJs/larva and 83.050 h., respectively. The lowest values of LD50 and LT50 for S. aciari were 71.192 IJs/larvae and 97.921 h., respectively. Overall, H. bacteriophora displayed more virulence toward O. obesus and A. ipsilon than S. aciari. Conclusion Both native EPNs were found effective against O. obesus and A. ipsilon. However, H. bacteriophora was more virulent toward O. obesus and A. ipsilon than S. aciari under the laboratory conditions.

Characterisation of the complete mitochondrial genome of the insect-parasitic nematode Heterorhabditis bacteriophora: an idiosyncratic gene order and the presence of multiple long non-coding regions

Summary We present here the complete mtDNA genome (mitogenome) of Heterorhabditis bacteriophora, an important biological control agent of soil-dwelling insect pests in agriculture and horticulture. This is the first description of a mitogenome for a member of the family Heterorhabditidae. The genome contains the typical chromadorean complement of 12 protein-coding genes, 22 tRNA genes and two rRNA genes. All genes are transcribed in the same direction and have a nucleotide composition high in A and T. For the entire genome, the nucleotide contents are 47.02% (T), 28.81% (A), 16.10% (G), 8.08% (C) and 75.83% (AT). Heterorhabditis bacteriophora has a unique, idiosyncratic gene arrangement. It differs from that of Caenorhabditis elegans in having a block of seven genes: trnQ-trnF-cytb-trnL1-cox3-trnT-nad4 translocated to a position between nad3 and nad5, as well as having a change in the position of the four tRNA block gene cluster, trnC-trnM-trnD-trnG, where trnC and trnM have switched places and trnD and trnG have translocated between nad4 and nad5 genes. The H. bacteriophora mitogenome is 18 128 bp long, and thus is ca 4 kb larger than the mitogenomes of most chromadoreans. This relatively large genome is due to the presence of five non-coding regions (NCR): NCR1 (114 bp), NCR2 (159 bp), NCR3 (498 bp), NCR4 (1917 bp) and NCR5 (2154 bp), which make up 26.7% of the genome. The NCR5 had the highest A + T content of 83.47% indicating that this region is the likely AT-rich control region. The complete 498 bp NCR3 sequence is duplicated in NCR4 and in NCR5 (the putative AT-rich control region). Such an organisation has not been reported previously in nematode mtDNA.

Efficacy of some native entomopathogenic nematodes against the alfalfa weevil, Hypera postica (Gyllenhal) (Coleoptera: Curculionidae), and the lucerne beetle, Gonioctena fornicata (Brüggemann) (Coleoptera: Chrysomelidae), adults under laboratory conditions

Background Entomopathogenic nematodes (EPNs) have more important role in biological control of economic insect pests. The effect of native EPNs on adults of the lucerne beetle, Gonioctena fornicata (Brüggemann, 1873) (Coleoptera: Chrysomelidae), and the alfalfa weevil, Hypera postica (Gyllenhal, 1813) (Coleoptera: Curculionidae), which are important alfalfa pests in Turkey and around the world, was investigated. Results Dose-mortality assays were carried out with 5 isolates [Steinernema carpocapsae (Weiser, 1955) (Nematoda: Steinernematidae) (Black sea isolate), S. feltiae Filipjev, 1934 (isolate 09-31), Heterorhabditis bacteriophora Poinar, 1976 (Nematoda: Heterorhabditidae) (isolate 09-43), H. bacteriophora Tokat-Songut, and S. carpocapsae Tokat-Ulas] using doses of 500, 1000, and 2000 IJs ml−1 under the laboratory conditions. Studies showed that all isolates had an effect 90% and more at 2000 IJs ml−1 and at the end of 112 h [except, H. bacteriophora (isolate 09-43) and H. bacteriophora Tokat-Songut isolates against H. postica]. In addition, LT30, LT50, and LT90 values at 1000 IJs ml−1 were determined. Conclusions According to the results, G. fornicata adults were susceptible to all isolates tested in the study and H. postica adults were susceptible to the isolates S. carpocapsae (Black sea isolate), S. feltiae (isolate 09-31), and S. carpocapsae Tokat-Ulas. This is the first study conducted in Turkey for the virulence of EPNs against G. fornicata and H. postica.