Molecular Epidemiology of Malarial Parasite From Punjab, Pakistan and its Control Through Plants Based Extracts and Nanoparticles

Pakistan, with 35% of the country’s landmass is classified as endemic for malaria and sixty percent population lives in such regions. Epidemiologically, Pakistan is considered as a moderate malaria endemic country where outbreak of malaria in different areas is varied in accordance to its population and temperature. The present study was based upon the survey of various government and private hospitals in ten cities of Northern and Southern Punjab and to find out the spatiotemporal pattern of microscopically positive cases of malarial infection in 2015. The 925 microscopically malaria positive blood samples were evaluated for distribution among the study area, seasonal variations, age groups and gender. However the suspected cases during the research period were 16075. The parasite species were identified both microscopically and on molecular basis. The Phylogenetic relationship of Plasmodium species was determined by DNA sequence analysis using software MEGA vs 7. The area wise distribution of malarial infection revealed that out of 925 microscopically positive cases, 364 cases were recruited from Northern Punjab with the highest from Rawalpindi (n=93, 25.5%) and the lowest from Chakwal ( (n=58, 15.9%). However 561 cases were observed from Southern Punjab with highest values from Rajanpur (n=120, 21.4%) and lowest one from Multan and Rahim Yar Khan i.e. (n=104, 18%). The slide positivity rate, Annual Parasite Prevalence and Annual Blood Examination Rate (in all recruited cities of both zones of Punjab were found to be 5.75%, 0.12 per 1000 population and 0.22% respectively. Seasonal variation of malarial prevalence revealed that it began to increase in the spring, peak in summer and Post monsoon, while showed minimum cases in winter. The age wise prevalence of malaria in 2015 revealed that it was highest among the young ones (1-20 years) while lowest cases were observed among people at the age of (41-60 years). However the gender wise prevalence indicated that the higher cases in males as compared to females and no age to gender wise relationship was observed. Microscopic and molecular diagnosis of malarial parasites revealed the occurrence of only two species of Plasmodium i.e. P. vivax and P. falciparum. The microscopic studies of 925 cases indicated the high abundance of P. vivax 16.7% (n=617) followed by P. falciparum 219 (23.7%) in the blood of recruited cases however 89 (9.62%) cases showed the mixed infection. Molecular analysis also revealed the same trend i.e. P vivax showed the higher occurrence followed by P. falciparum and the mixed infection. The overall prevalence of treatment seeking patients in all recruited cities of Punjab was 4.88%. However the comparison of Northern and Southern Punjab indicated that the prevalence was higher in Southern Punjab (5.54%) as compared to the Northern Punjab (4.01%). The overall results indicated that malarial infection is most prevalent in the Southern Punjab as compared to Northern Punjab and it is most prevalent during summer and moonsoon. Moreover people under the age of 20 were most susceptible and over all P. vivax showed the higher occurrence in the patients as compared to other Plasmodium species.