Isolation and Identification of Culturable Gut Microbiota in the Larval Stage of Lesser Mealworm (<i>Alphitobius diaperinus</i>)
The highly prevalent pest <i>Alphitobius diaperinus</i> (Coleoptera: Tenebrionidae) causes significant structural damage in poultry farms. Despite previous investigations on its carriage of pathogenic microorganisms, our understanding of its microbiome remains limited. This study aimed t...
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2023-11-01
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| author | Gisele Ivonne Antonuccio Diego Herman Sauka |
| author_facet | Gisele Ivonne Antonuccio Diego Herman Sauka |
| author_sort | Gisele Ivonne Antonuccio |
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| description | The highly prevalent pest <i>Alphitobius diaperinus</i> (Coleoptera: Tenebrionidae) causes significant structural damage in poultry farms. Despite previous investigations on its carriage of pathogenic microorganisms, our understanding of its microbiome remains limited. This study aimed to analyze the diversity of culturable gut microbiota in <i>A. diaperinus</i> obtained from laboratory breeding. Fifteen seventh instar larvae underwent a 24-h starvation period, followed by surface disinfection. Dissected midguts were homogenized and plated on nutrient agar (NA), brain heart infusion agar (BHI), and Bacillus cereus agar (BC). The cultured isolates were subjected to gram staining, phylogenetic analysis, biochemical property evaluation, and metabolic activity assessment. Bacterial counts were higher in BHI (2.51 × 10<sup>5</sup> CFU/gut) than in NA (2.25 × 10<sup>5</sup> CFU/gut), possibly due to nutrient richness. NA exhibited a dominant colony morphology of gram-negative bacilli, while BHI displayed additional distinct colonies of gram-positive cocci. Surprisingly, yeast-like colonies were observed on BC plates. Based on 16S rRNA gene sequences, eight bacterial isolates were identified as <i>Enterobacter</i> sp., and two as <i>Staphylococcus</i> sp. Using RNA gene ITS region sequences, two yeast isolates were identified as <i>Debaryomyces</i> sp. and <i>Hyphopichia</i> sp. A preliminary species-level identification of bacteria (<i>Enterobacter cloacae</i>, <i>Staphylococcus gallinarum</i>, and <i>Staphylococcus succinus</i>) was achieved using API systems and complementary biochemical tests. Discrepancies between phylogenetic analysis and phenotypic data suggest the potential existence of new species or subspecies. Further comprehensive studies are required to confirm this hypothesis. |
| format | Article |
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| publishDate | 2023-11-01 |
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| spelling | doaj-art-94f6201f3e31474cbcd58ee63a4214152025-08-20T02:00:35ZengMDPI AGBiology and Life Sciences Forum2673-99762023-11-013111210.3390/ECM2023-16465Isolation and Identification of Culturable Gut Microbiota in the Larval Stage of Lesser Mealworm (<i>Alphitobius diaperinus</i>)Gisele Ivonne Antonuccio0Diego Herman Sauka1Instituto de Microbiología y Zoología Agrícola (IMYZA), Instituto Nacional de Tecnología Agropecuaria (INTA), Buenos Aires 1686, ArgentinaInstituto de Microbiología y Zoología Agrícola (IMYZA), Instituto Nacional de Tecnología Agropecuaria (INTA), Buenos Aires 1686, ArgentinaThe highly prevalent pest <i>Alphitobius diaperinus</i> (Coleoptera: Tenebrionidae) causes significant structural damage in poultry farms. Despite previous investigations on its carriage of pathogenic microorganisms, our understanding of its microbiome remains limited. This study aimed to analyze the diversity of culturable gut microbiota in <i>A. diaperinus</i> obtained from laboratory breeding. Fifteen seventh instar larvae underwent a 24-h starvation period, followed by surface disinfection. Dissected midguts were homogenized and plated on nutrient agar (NA), brain heart infusion agar (BHI), and Bacillus cereus agar (BC). The cultured isolates were subjected to gram staining, phylogenetic analysis, biochemical property evaluation, and metabolic activity assessment. Bacterial counts were higher in BHI (2.51 × 10<sup>5</sup> CFU/gut) than in NA (2.25 × 10<sup>5</sup> CFU/gut), possibly due to nutrient richness. NA exhibited a dominant colony morphology of gram-negative bacilli, while BHI displayed additional distinct colonies of gram-positive cocci. Surprisingly, yeast-like colonies were observed on BC plates. Based on 16S rRNA gene sequences, eight bacterial isolates were identified as <i>Enterobacter</i> sp., and two as <i>Staphylococcus</i> sp. Using RNA gene ITS region sequences, two yeast isolates were identified as <i>Debaryomyces</i> sp. and <i>Hyphopichia</i> sp. A preliminary species-level identification of bacteria (<i>Enterobacter cloacae</i>, <i>Staphylococcus gallinarum</i>, and <i>Staphylococcus succinus</i>) was achieved using API systems and complementary biochemical tests. Discrepancies between phylogenetic analysis and phenotypic data suggest the potential existence of new species or subspecies. Further comprehensive studies are required to confirm this hypothesis.https://www.mdpi.com/2673-9976/31/1/12<i>Alphitobius diaperinus</i>gut microbiotaculturable microorganismsbacterial diversityyeast |
| spellingShingle | Gisele Ivonne Antonuccio Diego Herman Sauka Isolation and Identification of Culturable Gut Microbiota in the Larval Stage of Lesser Mealworm (<i>Alphitobius diaperinus</i>) Biology and Life Sciences Forum <i>Alphitobius diaperinus</i> gut microbiota culturable microorganisms bacterial diversity yeast |
| title | Isolation and Identification of Culturable Gut Microbiota in the Larval Stage of Lesser Mealworm (<i>Alphitobius diaperinus</i>) |
| title_full | Isolation and Identification of Culturable Gut Microbiota in the Larval Stage of Lesser Mealworm (<i>Alphitobius diaperinus</i>) |
| title_fullStr | Isolation and Identification of Culturable Gut Microbiota in the Larval Stage of Lesser Mealworm (<i>Alphitobius diaperinus</i>) |
| title_full_unstemmed | Isolation and Identification of Culturable Gut Microbiota in the Larval Stage of Lesser Mealworm (<i>Alphitobius diaperinus</i>) |
| title_short | Isolation and Identification of Culturable Gut Microbiota in the Larval Stage of Lesser Mealworm (<i>Alphitobius diaperinus</i>) |
| title_sort | isolation and identification of culturable gut microbiota in the larval stage of lesser mealworm i alphitobius diaperinus i |
| topic | <i>Alphitobius diaperinus</i> gut microbiota culturable microorganisms bacterial diversity yeast |
| url | https://www.mdpi.com/2673-9976/31/1/12 |
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