Abdul Haleem Shah

Characterization of gut bacterial flora of Apis mellifera from north-west Pakistan

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.

Controlling honeybee pathogen by using neem and Barbaka plant extracts

The honeybees (Apis mellifera) population is declining. The involved causes may be pathogens (mites, viruses and bacteria) and parasites, due to honeybees compromised immune system, leading to various bee-associated infections. Therefore, the present study assessed the comparative efficacy of plant extracts, including neem (Azadirachta indica) and Barbaka (Vitex trifolia) against gut bacteria and ectoparasitic mite Varroa destructor of honeybee A. mellifera (Hymenoptera: Apidae). The in vitro activities of the plant extracts were determined by using standard methods against five bee gut bacterial isolates, including the well-known bee pathogenic bacteria Paenibacillus larvae. Miticides were also assessed in field against honeybee mites. The obtained results from the phytochemical screening of Barbaka and neem extracts efficiency showed inhibitory zones with diameters of 23 mm with 60 mg/mL against P. larvae and 14 mm with 60 mg/mL against Escherichia coli, respectively. None of the extracts proved to be effective against Salmonella enterica and the neem extract showed intermediate activity against Bacillus subtilis and Staphylococcus hominis. Likewise, Barbaka plant extracts were not effective against B. subtilis. Similarly, the relative treatment efficacies of neem and Barbaka extracts, together with conventional miticides against honeybee Varroa mites, varied significantly. However, the effect of Barbaka and neem extracts on the mite-infested colonies was lower than the effect of other treatments, but it was also higher than in the control colonies. This study concluded that Barbaka and neem extracts have antibacterial and miticidal activity and are reasonably safe. However, more trials have to be conducted, in order to validate these results.

Prevalence of American foul brood disease of honeybee in north-west Pakistan

American foul brood (AFB) disease is a deleterious bacterial disease worldwide, caused by spore forming bacterium Peanibacillus larvae that affects honeybee larvae and causes a significant decrease in the honeybee population. Following symptomatical and bacteriological approaches combined with 16S rDNA sequencing, an assessment has been made to evaluate the presence of AFB disease in North-West Pakistan as no record for bee-associated bacterial disease from Pakistan is available. A total of 1276 samples from 1520 bee colonies (15 apiaries) were collected, of which 476 samples (37.30%) were found with symptoms of AFB. Biochemical and 16S rDNA analysis indicated that all these farms have Peanibacillus larvae infection. It is concluded that the prevalence of AFB bacterial disease to such an extent in these regions of Pakistan will devastate the apicultural industries in a large scale across the country.

PCR/RFLP-Based Analysis of Genetically Distinct Plasmodium vivax Population of Pvmsp-3α and Pvmsp-3β genes in Pakistan

BackgroundPlasmodium vivax is one of the widespread human malarial parasites accounting for 75% of malaria epidemics. However, there is no baseline information about the status and nature of genetic variation of Plasmodium species circulating in various parts of Pakistan. The present study was aimed at observing the molecular epidemiology and genetic variation of Plasmodium vivax by analysing its merozoite surface protein-3α (msp-3α) and merozoite surface protein-3β (msp-3β) genes, by using suballele, species-specific, combined nested PCR/RFLP detection techniques.MethodsA total of 230 blood samples from suspected subjects tested slide positive for vivax malaria were collected from Punjab, Sindh, Khyber Pakhtunkhwa, and Balochistan during the period May 2012 to December 2013. Combined nested PCR/RFLP technique was conducted using Pvmsp-3α and Pvmsp-3β genetic markers to detect extent of genetic variation in clinical isolates of P. vivax in the studied areas of Pakistan.ResultsBy PCR, P. vivax, 202/230 (87.82%), was found to be widely distributed in the studied areas. PCR/RFLP analysis showed a high range of allelic variations for both msp-3α and msp-3β genetic markers of P. vivax, i.e., 21 alleles for msp-3α and 19 for msp-3β. Statistically a significant difference (p ≤ 0.05) was observed in the genetic diversity of the suballelic variants of msp-3α and msp-3β genes of P. vivax.ConclusionIt is concluded that P. vivax populations are highly polymorphic and diverse allelic variants of Pvmsp-3α and Pvmsp-3β are present in Pakistan.