Isolation of Hermetia illucens larvae core gut microbiota by two different cultivation strategies.

Hermetia illucens larvae (black soldier fly larvae, BSFL) convert efficiently organic waste to high quality biomass. To gain knowledge on the specific functions of gut microbes in this process it is a prerequisite to culture members of the core gut microbiota. Two different cultivation strategies were applied here for this purpose, a dilution-to-extinction cultivation and direct plating using six different media to culture aerobic heterotrophic bacteria. A total of 341 isolates were obtained by the dilution-to-extinction cultivation and 138 isolates by direct plating from guts of BSFL reared on chicken feed. Bacterial isolates were phylogenetically identified at the genus level by 16S rRNA gene sequencing (phylotyping) and differentiated at the strain level by genomic fingerprinting (genotyping). The main proportion of isolates was assigned to Proteobacteria, Firmicutes (Bacilli), and Actinobacteria. Predominant genera discussed in literature as member of a potential BSFL core gut microbiota, Providencia, Proteus, Morganella, Enterococcus, Bacillus, and members of the family Enterobacteriaceae, were isolated. A high intra-phylotype diversity was obtained by genomic fingerprinting which was especially enhanced by the dilution-to-extinction cultivation. This study showed that the application of different cultivation strategies including a dilution-to-extinction cultivation helps to culture a higher diversity of the BSFL gut microbiota and that genomic fingerprinting gives a better picture on the genetic diversity of cultured bacteria which cannot be covered by a 16S rRNA gene sequence based identification alone.

The gut and feed residue microbiota changing during the rearing of Hermetia illucens larvae.

Larvae of Hermetia illucens, commonly known as black soldier fly, efficiently convert organic waste into nutrient-rich supplements for different applications. Here we performed a preliminary experiment to investigate the dynamics of the H. illucens gut microbiota and changes in the composition of the bacterial community in the residue of the larval feed during rearing. We furthermore quantified the presence of antibiotic resistance and disinfectant genes in the gut and feed microbiota during the rearing process to elucidate if rearing leads to a reduction, increase, and/or transfer of resistance genes from the feed to larvae and vice versa. We found that the gut and feed residue bacterial communities were distinct throughout the rearing process. The gut microbiome remained more stable compared to the feed residue microbiome varying in both bacterial abundance and community structure during rearing. Antibiotic-resistance genes were present in both, gut and feed residues, with a significant increase in pupae and residue samples taken at the end of the rearing process. Disinfectant-resistance genes were present in the feed residue and even increased during the rearing process but were not transferred to the gut microbiome. We conclude that H. illucens larvae have a stable gut microbiome that does not change significantly over the course of larval development, whereas bacterial communities in the feed residue are strongly affected by rearing. If the presence of antibiotics and disinfectants during rearing, can promote the spread of antibiotic/disinfectant-resistance genes among feed and larvae needs to be evaluated in further experiments.

Genome sequencing of three bacteria associated to black band disease from a Colombian reef-building coral.

We announce the draft genome sequence of three Gram-negative bacteria isolated from coral tissues affected with the black band disease (BBD), identified with the NCBI's Assembly Database accession numbers: MBQF, MAYB and MBQE. These genome drafts constitute an useful tool for the characterisation of these bacteria and for the understanding of the relationship between the microbial consortia associated with the disease and the onset and progression of the pathology.