![]() ![]() Individual colonies were subcultured twice to ensure purity (Kuzina et al. Different colony types of bacteria were selected and purified on nutrient agar plates by subculturing. After incubation, the plates were examined and bacterial colonies that were similar in terms of colony and color morphology were selected. The plates were incubated at 25–36 ☌ for 24–48 h. Then, 100 μl hemolymph suspension, taken from the hemocoel, was spread on nutrient agar plates. Sterile 1 ml syringes were used to insert into the hemocoel of the insect. After surface sterilization, considering aseptic conditions samples were washed by a sterilized water. Before the isolation process, the dead larvae were individually placed into 70% ethanol and gently shaken for 3 min (Yaman et al. melolontha larvae suspected of bacterial symptoms after a macroscopic examination were used for bacterial isolation (Thiery and Frachon 1997). pityocampa larvae were collected from pine trees in Sinop province, Turkey, brought to the laboratory, and fed on fresh pine needles. Lymantria dispar larvae were collected from İzmir province, Turkey, brought to the laboratory, and fed on apple leaves. micans larvae were obtained from the forest fields near Ordu, Turkey their cultures were maintained in a laboratory set up by the Turkish Ministry of Forestry in Ordu province. 2010), Thaumetopoea pityocampa, and Lymantria dispar, were used. For bioassay tests, three economic insect pests, namely Dendroctonus micans (Yaman et al. melolontha larvae were collected from Ordu province, Turkey, targeting isolate entomopathogenic bacteria. melolontha larvae were presented and their potentials on three economic insect pests were documented. In this study, isolation and characterization of non-spore-forming bacteria from M. It is a soil-inhabiting insect pest at the larval stage and possible target for entomopathogenic bacteria in soil. The European cockchafer, Melolontha melolontha ( Linnaeus, 1758) (Coleoptera: Scarabaeidae) causes significant economic damage. Scarabs are infected by a number of entomopathogens (Yaman et al. Numerous larvae of Scarabaeidae (Coleoptera) feed on plant materials in the soil, and many of them are known as important plant pests. While the entomopathogenic bacteria domain has been well represented by spore-forming bacteria, members of Bacillaceae family, non-spore-forming entomopathogenic bacteria have been also documented with insects (Ruiu et al. Soil plays an important role as a habitat for spore-forming and non-spore-forming entomopathogenic bacteria. Entomopathogenic bacteria are commonly isolated from host insects or soil (Thiery and Frachon 1997). Current goals include searching for entomopathogenic bacteria and determining their potentials (Egami et al. Entomopathogenic bacteria such as Bacillus thuringiensis are generally known as lower risk pesticides than chemicals. Among them, bacteria and their toxins are the most commercially microbial insecticides used successfully in biological control. The successful use of entomopathogens results in an alternative pest management for insect control. Entomopathogens have relative specificity and lower environmental impact. New pest management strategies tend to minimize the impact on the environment and non-target organisms (Ruiu et al. The isolated bacteria can be considered in integrated pest control programs. liquefaciens 45, 65.2, and 36% mortality rates on the larvae of D. Laboratory experiments, carried out to evaluate the virulence of these isolates, showed that all isolated bacteria had a pathogenic effect on the tested pests. ![]() The bacteria were identified as Enterobacter cloacae complex (isolate 1M), Serratia marcescens (isolate 3M), Pseudomonas aeruginosa (isolate 4M), Kocuria kristinae (isolate 5M), and Serratia liquefaciens (isolate 8M). All isolated bacteria were cultured and identified using VITEK bacterial identification systems and 16S rRNA gene sequence analysis. Their potential was tested against the three economic insect pests, the great spruce bark beetle, Dendroctonus micans Kugelann (Coleoptera: Curculionidae) the pine processionary, Thaumetopoea pityocampa (Lepidoptera: Thaumetopoeidae) and the gypsy moth, Lymantria dispar (Linn.) (Lepidoptera: Erebidae), to find an effective biological control agent. Five non-spore-forming bacteria were isolated from the European cockchafer, Melolontha melolontha ( Linnaeus, 1758) (Coleoptera: Scarabaeidae). ![]()
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