Actinomycetes, as one of the rhizosphere bacteria, also produce a wide range of hydrolytic exoenzymes (e.g. chitinases, cellulase, etc.), and are therefore primary contributors to the cycling of carbon in organic matter derived from fungi and plants. Because of the importance and potential growth advantages of these bacteria,

several studies have focused on the isolation and visualization see more of actively growing actinomycetes in the guts of beetles, termites and millipedes (Bignell et al., 1979; Gozev & Byzov, 2006; Scott et al., 2008). Previously, nonpathogenic microbiota associated with honeybees have mostly been examined using classical culture-based techniques, and chemotaxonomic characterization of the isolates, which have described a group of Gram-variable pleomorphic bacteria in honeybee guts but not in adequate detail (Gilliam, 1997). Although data from the latest pyrosequencing technology applied to honeybee gut microbiota are yet to be published, few metagenomic studies have revealed the presence of actinomycetes in this environment (Cox-Foster et al., 2007). Also, it is known that PCR amplification of bacterial 16S rRNA genes with universal primers could have dramatically underestimated the population

of high-GC Actinobacteria in a complex community (Stach et al., 2003). However, one culture-based report indicated that Streptomyces PR-171 nmr sometimes could become dominant in bee guts (Mohr & Tebbe, 2007). To our knowledge, no antibiotic-producing actinomycetes from the

guts of honeybees have ever been characterized, though Streptomyces are among the microorganisms found in honey (Snowdon & Cliver, 1996) and honey products have well-known antimicrobial properties (Kwakman et al., 2008). Honey has been a popular folk medicine for healing wound and soothing sore throat since ancient times. In this report, selective media were used to isolate actinomycetes from the digestive tract of adult honeybees. The antibiotic activities produced under laboratory conditions were evaluated against bee indigenous Bacillus strains, Escherichia coli and two drug-resistant human pathogens. One frequently encountered Vasopressin Receptor isolate identified as a species of Nocardiopsis was further characterized and the expression of an antibiotic biosynthetic gene was analyzed. Adult worker honeybees were collected from six locations, most of which have <10 isolated hives. Within 12 h of capture, bees were externally sterilized with 70–100% alcohol and dissected under sterile conditions. The digestive tracts, from crop to rectum, were pooled, lightly homogenized and suspended in saline and plated on selective agar plates. The gut contents from each bee were spread on one plate. To better investigate the actinomycete diversity in the complex microbial milieu of the insect gut, different selective media were used for the colony isolation.