Gut microbiota has been recognized to play a beneficial role in honey bees (Apis mellifera). Present study was designed to characterize the gut bacterial flora of honey bees in north-west Pakistan. Total 150 aerobic and facultative anaerobic bacteria from guts of 45 worker bees were characterized using biochemical assays and 16S rDNA sequencing followed by bioinformatics analysis. The gut isolates were classified into three bacterial phyla of Firmicutes (60%), Proteobacteria (26%) and Actinobacteria (14%). Most of the isolates belonged to genera and families of Staphylococcus, Bacillus, Enterococcus, Ochrobactrum, Sphingomonas, Ralstonia, Enterobacteriaceae, Corynebacterium and Micrococcineae. Many of these bacteria were tolerant to acidic environments and fermented sugars, hence considered beneficial gut inhabitants and involved the maintenance of a healthy microbiota. However, several opportunistic commensals that proliferate in the hive environment including members Staphylococcus haemolyticus group and Sphingomonas paucimobilis were also identified. This is the first report on bee gut microbiota from north-west Pakistan geographically situated at the crossroads of Indian subcontinent and central Asia.

We aimed to characterize the relationship of the oral microbiome with diabetes in Pakistan. Saliva samples were collected from diabetic patients (n = 49) and healthy individuals (n = 55). 16S metagenomics saliva was carried out by NGS technology. We observed that the phylum Firmicutes (p-value = 0.024 at 95% confidence interval) was significantly more abundant among diabetic patients than among the controls. We found that the abundance of phylum Actinobacteria did not significantly vary among both groups in contrast to a similar report from the USA (Long et al., 2017). On genus level, acidogenic bacteria Prevotella (p-value = 0.024) and Leptotrichia (p-value = 1.5 × 10 ⁻³ ); and aciduric bacteria Veillonella (p-value = 0.013) were found to be in higher abundance in diabetic patients. These bacteria are found in dental biofilm and involved in the metabolism of fermentable carbohydrates. Stratified analysis by gender revealed healthy and diabetic females to be more divergent. Abundance of Prevotella (p-value = 4.4 × 10-3) and Leptotrichia (p-value = 0.015) was significantly associated with male patients. A comparison of oral bacteriome between two groups revealed the dominance of acidogenic and aciduric bacteria in diabetics which suggested the involvement of these eubacteria in oral dysbacteriosis in diabetes mellitus.

Betel nut addiction is recognized as the causative agent of oral microbiome dysbiosis and other systematic disorders. A number of betel nut preparations containing ingredients such as slaked lime, catechu extract and tobacco are being commonly used particularly in South Asia. The underlying variations in the oral microbiome due to usage of betel nut preparations are poorly understood. We evaluated salivary microbiome in response to chewing of betel nut preparation(s). In order to assess the microbiome dynamics, metagenomic analysis of 16S rRNA gene (V3-V4 hypervariable region) from salivary bacteria in chewers of betel nut preparation (n = 16) and non-chewers (n = 55) was carried out by Greengenes and SILVA ribosomal sequence databases. It was observed that Gutka chewers demonstrated lower alpha diversity and number of bacterial genera than the non-chewers. Taxonomic assignment on phylum level revealed Firmicutes (p-value = 0.042 at 95% confidence interval) to be significantly more abundant in Gutka chewers in comparison with non-chewers. Beta diversity analysis at genus level by weighted unifrac distance matrices unveiled both groups to be divergent from each other. On the genus level, Veillonella (p-value = 0.015), Streptococcus (p-value = 0.026), Leptotrichia (p-value = 0.022) and Serratia (p-value = 0.022) species appeared to be significantly more abundant in Gutka chewers in comparison to non-chewers. The present study suggests salivary dysbiosis in response to gutka chewing and concludes that gutka chewers possess higher abundance of acidogenic and aciduric bacteria. This study contributes additional information regarding oral microbiome variations with response to gutka consumption.