Isolation and identification of bacteria from Thaumetopoea pityocampa Den. and Schiff. (Lep., Thaumetopoeidae) and determination of their biocontrol potential

World Journal of Microbiology and Biotechnology (Impact Factor: 1.78). 12/2008; 24(12):3005-3015. DOI: 10.1007/s11274-008-9845-9


The pine processionary moth Thaumetopoea pityocampa (Den. and Schiff.) is one of the most harmful insect pest for pine species in Mediterranean countries including Turkey. The
objective of the present study is to find a more effective and safe biological control agent against T. pityocampa. Thus, we investigated the bacterial flora of the pest insect, collected from the Middle Black Sea Region of Turkey from
2003 to 2004. Based on morphological, physiological, biochemical and molecular methods, 14 different bacterial isolates were
determined. The identified bacterial flora of T. pityocampa consisted of bacteria belonging to the Enterobacteriaceae (Tp1), Arthrobacter sp. (Tp2), Staphylococcus spp. (Tp3 and 10), Bacillus subtilis (Tp4), Serratia liquefaciens (Tp5), Bacillus thuringiensis subsp. morrisoni (Tp6 and 14), an acrystalliferous form Bacillus thuringiensis (Tp7), Staphylococcus cohnii (Tp8), Bacillus licheniformis (Tp9), Bacillus pumilus (Tp11), Brevibacterium sp. (Tp12) and Bacillus simplex (Tp13). After analysing the conclusions of conventional and molecular tests Tp1 (Enterobacteriaceae), Tp2 (Arthrobacter sp.) and Tp12 (Brevibacterium sp.) were assigned as novel bacterial species. Isolate Tp5 had a promising insecticidal effect on third instar larvae of
T. pityocampa (up to 70% mortality within 10days).

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    • "During their development, PPMs are susceptible to attack by a range of hymenopteran and dipteran parasitoid species, including parasitoids of egg, larvae and pupae (Biliotti, 1955; Du Merle, 1969; Tsankov et al., 1996; Schmidt et al., 1999; Lòpez-Sebastián et al., 2007; Bonsignore et al., 2011). PPMs are also attacked by invertebrate predators, including ants, beetles and spiders, as well as by vertebrates, such as birds, shrews and voles, and also by pathogens (Way et al., 1999; Battisti et al., 2000; Er et al., 2007; Ince et al., 2008; Hatice et al., 2009; Barbaro & Battisti, 2011). However, it has been difficult to quantify the effect of parasitoids on PPM developmental stages that occur underground, in particular, which species of parasitoids are involved and how they influence the PPM population dynamics. "
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    ABSTRACT: Parasitoids are significant enemies of many economically important insects and there is some evidence to suggest that their actions have a role in terminating the outbreaks of forest Lepidoptera populations. In this study, we examined the impact of parasitoids on the pupae of the pine processionary moth, and highlighted the presence of several parasitoid species for this developmental stage. A higher rate of parasitism was found when the pupal density in the soil was reduced, but the rate of parasitism was not influenced by pupal morphological traits or by the presence or absence of a cocoon around a pupa. Of the external factors examined, a delay in the time of descent of larvae from the trees had a positive effect on the level of parasitism. Observational data indicated that dipteran and hymenopteran were the most abundant parasitoids to emerge from moth pupae. Our study highlights the complexity of the parasitoid–host dynamics, and stresses the importance of carefully determining environmental effects on host–parasitoid relations.
    Bulletin of Entomological Research 06/2015; 105(5):1-8. DOI:10.1017/S0007485315000541 · 1.91 Impact Factor
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    • "Efforts have been increased to discover more effective and safe biological control against hazardous insects to replace potentially hazardous chemical agents (Ince et al. 2008). Moreover, investigating a new pathogen from various environments against a target pest is always desirable. "
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    ABSTRACT: Agrotis segetum Schiff. (Lepidoptera: Noctuidae) is one of the most serious pests of nearly all vegetables in Turkey. In this study, to find a more effective and safe biological control agent, the bacterial flora of Agrotis segetum were investigated in larvae collected from different populations in the vegetable fields of the Black Sea region of Turkey, and 9 bacterial isolates were cultured from these populations. Seven of these bacteria were determined and characterized at the species level and the rest were characterized at the genus level. Morphological, physiological, and biochemical characteristics of the bacterial isolates were determined by conventional and routine techniques. Characterization of the isolates was also performed using the VITEK 32 bacterial identification system. In addition, a 16S rRNA gene sequence was determined for these isolates to query against the NCBI genetic database and to construct a phylogenetic tree using closely related species. Consequently, the isolates were identified as Bacillus cereus (Ags1), Bacillus sp. (Ags2), Bacillus megaterium (Ags3), Enterobacter aerogenes (Ags4), Acinetobacter calcoaceticus (Ags5), Enterobacter sp. (Ags6), Pseudomonas putida (Ags7), Enterococcus gallinarum (Ags8), and Stenotrophomonas maltophilia (Ags9). All isolates were determined the first time from the flora of A. segetum. Furthermore, the insecticidal activities of the bacterial isolates were tested against A. segetum larvae, and Enterococcus gallinarum (Ags8) was found to cause the highest insecticidal activity (60%) 8 days after application. Agrotis segetum Schiff. (Lepidoptera: Noctuidae)'un bakterileri ve bu bakterilerin insektisidal etkileri üzerine yeni bir çalışma Özet: Agrotis segetum Schiff. (Lepidoptera: Noctuidae) Türkiye' de hemen hemen bütün sebzelerin en önemli zararlılarından birisidir. Bu çalışmada, daha etkili ve güvenli biyolojik mücadele etmeni bulmak için Agrotis segetum'un bakteriyal florası Karadeniz Bölgesi'ndeki sebze tarlarında farklı populasyonlardan toplanan larvalar üzerinde araştırıldı. Bu populasyonlardan dokuz bakteri izolatı kültüre alındı. Bu bakterilerden yedi tanesi tür seviyesinde, diğerleri ise cins seviyesinde tanımlandı ve karakterize edildi. Bakteriyal isolatların morfolojik, fizyolojik ve biyokimyasal özellikleri, rutin ve geleneksel teknikler kullanılarak belirlendi. İzolatların karakterizasyonu, VITEK 32 bakteriyal tanımlama sistemi kullanılarak da gerçekleştirildi. Buna ilaveten, bütün izolatların NCBI genetik veritabanında karşılaştırmalarını yapmak ve ilişkili türler ile filogenetik yakınlıklarını belirlemek için 16S rRNA gen sekans analizleri yapıldı. Sonuç olarak, izolatlar Bacillus cereus (Ags1), Bacillus sp. (Ags2), B. megaterium (Ags3), Enterobacter aerogenes (Ags4), Acinetobacter calcoaceticus (Ags5), Enterobacter sp. (Ags6), Pseudomonas putida (Ags7), Enterococcus gallinarum (Ags8) ve Stenotrophomonas maltophilia (Ags9) olarak tanımlandı. Bütün izolatlar A. segetum'un florasında ilk kez belirlendi. Ayrıca, bakteriyal izolatların insektisidal etkileri A. segetum larvalarına karşı test edildi. Uygulamadan 8 gün sonra Enterococcus gallinarum (Ags8)'un en yüksek aktiviteye (%60) sebep olduğu bulundu.
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    ABSTRACT: The pine processionary moth Thaumetopoea pityocampa Den. and Schiff. (Lep., Thaumetopoeidae) is one of the most harmful insect pests for pine species in Mediterranean countries including Turkey. Two Bacillus thuringiensis isolates obtained from T. pityocampa were identified and characterized in terms of crystal shape using electron microscopy, SDS–PAGE analysis, cry gene contents, H-serotype and insecticidal activity. Examination by a scanning electron microscope showed that Tp6 and Tp14 isolates have flat square and bipyramidal crystal shapes, respectively. PCR analysis showed that Tp6 contains cry3 gene and Tp14 isolate contains cry1 and cry2 genes. On the other hand, the presence of Cry3 and Cry1 proteins were confirmed by observation of approximately 65- and 130-kDa proteins by SDS–PAGE in Tp6 and Tp14 isolates, respectively. According to H-serotype results, these isolates were identified as Bacillus thuringiensis ssp. morrisoni (H8a8b). Toxicity tests were performed against six insect species belonging to Lepidoptera and Coleoptera. The highest insecticidal activity was 100% for Tp6 isolate on larvae of Agelastica alni and Leptinotarsadecemlineata and 100% for Tp14 isolate on larvae of Malacosoma neustria. Our results indicate that isolates Tp6 and Tp14 may be valuable biological control agents for various coleopteran and lepidopteran pests.
    Biocontrol Science and Technology 05/2009; 19(5):475-484. DOI:10.1080/09583150902836377 · 0.94 Impact Factor
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