HOST SPECIES AND FOREST HABITAT SHAPE THE EXTERNAL BACTERIAL MICROBIOMES OF FOREST BLOWFLIES (DIPTERA: CALLIPHORIDAE) IN MALAYSIA

Ahmad Azani Othman, Atikah Insyirah Azizi, Zolkapli Eshak, Mohammad Adam Adman, Zulkiflee Abd Latif, Muhamad Azizi Mustapa, Mohamad Faiz Foong Abdullah, Azwandi Ahmad

Abstract


Necrophagous flies harbour diverse bacterial communities that reflect their interactions with decomposing substrates and the surrounding environment. This study characterised the external bacterial microbiomes of two forest-associated blowflies, Chrysomya villeneuvi Patton and Phumosia promittens Walker, across two urban forest fragments (Ayer Hitam Forest Reserve and Bukit Rupa UKM Forest Reserve) and one continuous rainforest (Kuala Keniam Rainforest, Pahang National Park). Sampling was conducted using modified hanging traps baited with 72-hour decomposed mackerel fish. In total, 74 Ch. villeneuvi and 158 P. promittens individuals were captured, and external microbiomes from five samples per species at each location were aseptically isolated and characterised using 16S rRNA gene amplicon sequencing. Principal coordinates analysis demonstrated clear separation between the microbiomes of Ch. villeneuvi and P. promittens in Kuala Keniam, whereas samples from Ayer Hitam and UKM exhibited greater overlap. PERMANOVA revealed a significant effect of host species identity on bacterial community composition across rainforest habitats (P<0.05). Core microbiome analysis indicated that Ch. villeneuvi uniquely contained Klebsiella, while P. promittens possessed several unique taxa, including Orbus, Streptococcus, Kaistella, Ligilactobacillus, Staphylococcus, and Orbus sasakiae. Microbiome composition also varied among forest habitat contexts, with Ch. villeneuvi from Kuala Keniam forming more distinct clusters than populations from fragmented forests, while P. promittens showed greater overlap among locations. PERMANOVA further indicated a significant effect of forest type on bacterial community composition in both species (P<0.05). Collectively, these findings provide evidence that host species identity is a key factor shaping the external bacterial microbiomes of forest-associated blowflies. Additionally, habitat context, particularly differences between continuous and fragmented rainforest ecosystems, may contribute to microbiome variation. These patterns are considered preliminary and require further investigation using larger sample sizes and synchronised sampling designs to minimise potential temporal effects. The findings provide baseline information for future studies exploring blowfly-associated microbiomes as potential indicators of tropical rainforest ecosystem health.


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