New Gammaproteobacteria were isolated from 3rd stage fly larvae of the parasitic fly Wohlfahrtia magnifica. Phylogenetic analysis of the new isolates showed that these bacteria belong to a distinct lineage close to Ignatzschineria larvae, which was originally isolated from the same species of fly. The low similarity values in 16S rRNA gene sequences (93.8-94.8 %), and differences in fatty acid profiles, RiboPrint patterns, MALDI-TOF mass spectra of cell extracts, and physiological and biochemical characteristics differentiate the isolates from the type strain of Ignatzschineria larvae (DSM 13226 T ), and indicate that our isolates represent a new genus within the Gammaproteobacteria. The major isoprenoid quinone of the strains is Q8, the major fatty acids are C 18 : 1 and C 14 : 0 , and the predominant polar lipids are phosphatidylglycerol, phosphatidylethanolamine and phosphatidylserine. The G+C content of the DNA of the type strain is 44.3 mol%. The name Wohlfahrtiimonas chitiniclastica gen. nov., sp. nov., is proposed for this novel genus and species. The type strain is S5 T (5DSM 18708Myiasis is an infestation of vertebrate animals with dipterous larvae. The obligate parasitic fly Wohlfahrtia magnifica develops only in live vertebrates and is a serious pest of livestock of Eastern Europe, the Mediterranean and Middle Asia (Nedelchev, 1988;Ruiz-Martinez et al., 1991;Hall & Wall, 1995;Farkas et al., 1997;Tó th et al., 1998 (Homonnay, 2006). The aim of the present work was to determine the taxonomic position of strains S5T and E43 and characterize them by using a polyphasic approach.
Strain S5T was isolated from the homogenate of 3rd stage larvae of Wohlfahrtia magnifica (Diptera: Sarcophagidae), collected from a primary laboratory culture of the fly. Three larvae were taken out of the cultivation medium, washed three times in sterile distilled water and homogenized in a sterile glass mortar. Strain E43 was isolated from the aseptically excised foregut of a 3rd stage maggot of Wohlfahrtia magnifica, collected from the infested vulval wound of a Romney breed sheep at Mezőfalva State Farm, Hungary (18 u 409 E, 46 u 509 N), in 2002. It was washed three times in sterile 0.025 M sodium phosphate buffer solution (pH 6.8) and the foregut was dissected under a preparatory microscope under aseptic conditions. Bacterial strains S5T and E43 were isolated and maintained on King B agar medium (King et al., 1954).Colony morphology of strains was tested on King B agar medium by direct observation of single colonies. Cell morphology and motility were studied in native preparations Abbreviations: PE, phosphatidylethanolamine; PG, phosphatidylglycerol; PS, phosphatidylserine.The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain S5T is AM397063.A transmission electron micrograph, a dendrogram based on MALDI-TOF data and the polar lipid pattern of strain S5