Five bacterial strains were isolated from the hemocoel of the greater wax moth larvae (Galleria mellonella) infected with the entomopathogenic nematodes: Heterorhabditis bacteriophora HP88, Heterorhabditis indicus RM1 and Heterorhabditis sp (S1), Steinernema abbasi and Steinernema sp. (S II). Strains were identified as Photorhabdus luminescens HRM1, P. luminescens HS1, P. luminescens HP88, Xenorhabdus indica and X. nematophila ATTC19061 using 16S rDNA sequence analysis. To reveal the genetic diversity among these strains, three molecular markers (RAPD, ISSR and SRAP) were employed. RAPD analysis showed 73.8 and 54.5 polymorphism percentages for the Photorhabdus and Xenorhabdus strains, respectively. ISSR analysis resulted in 70.1 and 75.2 polymorphism percentages among the Photorhabdus and Xenorhabdus strains, respectively. The SRAP analysis indicated that 75.6 and 61.2% genetic polymorphism was detected among Photorhabdus and Xenorhabdus strains, respectively. The cluster analysis grouped the three Photorhabdus strains together in one cluster and the two Xenorhabdus strains together in another cluster indicating the phylogenetic relationships among them. The genotype-specific markers detected from the three molecular markers (RAPD, ISSR and SRAP) were sufficient to distinguish between the different bacterial strains tested and can be used in the future IBM program that could be built on the use of these strains.
Biological control of hard ticks, Hyalomma dromedarii (Acari: Ixodidae) using entomopathogenic nematode of two families; Heterorhabditidae and Steinernematidae was studied. The protective effect of controlled ticks including haemolymph and haemocytes against these biological agents were also investigated. It was found that heterorhabditid strains cause a higher effect in biological control of engorged female H. dromedarii than those of stienernematid strains. It induced mortality rates ranged from 12-92% versus 4-88% for stienernematid strains. It was also found that these entomopathogenic nematodes can not reproduce within the exposed ticks. SDS-PAGE of proteins extracted from midguts and salivary glands infected with 4000 IJs tick(-1) separated 21 and 25 protein bands versus 13 and 19 protein bands from non-infected ones, respectively. It was concluded that entomopathogenic nematodes of family Heterorhabditidae proved to have a potential acaricidal effect in the control of hard ticks. Moreover, the controlled ticks released unknown proteins in their haemolymph that may promote the haemocytes to phagocyte the nematodes as a type of defense mechanism.
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