AUTHOR=Milanović Vesna , Osimani Andrea , Roncolini Andrea , Garofalo Cristiana , Aquilanti Lucia , Pasquini Marina , Tavoletti Stefano , Vignaroli Carla , Canonico Laura , Ciani Maurizio , Clementi Francesca 

TITLE=Investigation of the Dominant Microbiota in Ready-to-Eat Grasshoppers and Mealworms and Quantification of Carbapenem Resistance Genes by qPCR

JOURNAL=Frontiers in Microbiology

VOLUME=9

YEAR=2018

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2018.03036

DOI=10.3389/fmicb.2018.03036

ISSN=1664-302X

ABSTRACT=<p>In this study, 30 samples of processed edible mealworms (<italic>Tenebrio molitor</italic> L.) and 30 samples of grasshoppers (<italic>Locusta migratoria migratorioides</italic>) were obtained from producers located in Europe (Belgium and the Netherlands) and Asia (Thailand) and subjected to PCR-DGGE analyses. The PCR-DGGE analyses showed that species in the genus <italic>Staphylococcus</italic> were predominant in the samples of mealworms from Belgium and grasshoppers from the Netherlands; species in the genus <italic>Bacillus</italic> were detected in the samples of mealworms and grasshoppers from Thailand. Moreover, <italic>Weissella cibaria/confusa</italic>/spp. was found in grasshoppers from Belgium. Since data concerning the role of novel foods such as edible insects in the dissemination of carbapenem resistance are currently lacking, the quantification of five carbapenemase encoding genes (<italic>bla</italic><sub>NDM−1</sub>, <italic>bla</italic><sub>VIM</sub>, <italic>bla</italic><sub>GES</sub>, <italic>bla</italic><sub>OXA−48</sub>, and <italic>bla</italic><sub>KPC</sub>) by qPCR was also carried out in all the samples under study. The genes coding for GES and KPC were not detected in the analyzed samples. A very low frequency of <italic>bla</italic><sub>OXA−48</sub> (3%) and <italic>bla</italic><sub>NDM−1</sub> (10%) genes was detected among mealworms. In contrast, grasshoppers were characterized by a high incidence of the genes for OXA-48 and NDM-1, accounting for 57 and 27% of the overall grasshopper samples, respectively. The <italic>bla</italic><sub>VIM</sub> gene was detected exclusively in two grasshopper samples from Thailand, showing only 7% positivity. The analysis of variance showed that all the effects (producers, species, and producers × species) were statistically significant for <italic>bla</italic><sub>NDM−1</sub>, whereas for <italic>bla</italic><sub>OXA−48</sub> and <italic>bla</italic><sub>VIM</sub>, no significant effects were detected for the same source of variation. Further studies are necessary to assess the possible role of edible insects as reservoirs for the resistance to carbapenems and to understand the correlation with the insect microbiota. Furthermore, an intensified surveillance plan examining the occurrence of carbapenemase encoding genes in the food chain and in environmental compartments is needed for a proper risk assessment. In such a context, the appropriate use of antimicrobials represents the main preventive action that should always be applied.</p>