Akhtar Rasool

Cutaneous Leishmaniasis (CL) is a vector-borne disease caused by Leishmania species, which is responsible for anthroponotic and zoonotic CL in Pakistan. Due to the unavailability of reports on the molecular epidemiology and risk factors assessment of CL in Khushab, Punjab, Pakistan, this study aimed to investigate the incidence rate of CL, associated risk factors, and molecular identification of Leishmania species in this region. The clinical and demographic data of 423 suspected CL patients were collected. Their skin scrapings and blood samples were analyzed microscopically for amastigote detection and for genomic DNA extraction, respectively. A partial fragment (~ 400 bp) of ITS1 gene was amplified followed by restriction fragment length polymorphism (RFLP) assay and sequencing for species identification. Meteorological data were obtained from the Pakistan Meteorological Department, Lahore. Statistical analysis was conducted using Univariate and multivariate regression models using SPSS. Microscopy confirmed 41.8% of the cases, while PCR detected Leishmania DNA in 33.8% of the samples. CL infection was more common in females (57%) and in the < 1–20 age group (54.8%). The majority of the CL lesions were dry (63.2%), single (69.4%), predominantly affecting the facial region (46.3%), occurred with a duration of less than 2 months (75.7%), and were less than 1 cm in size (62.7%). Cases were highest in January (19.3%) and lowest in June (2.6%). The average temperature (22–27 °C) and reduced rainfall (4 mm) were key factors for sand-fly breeding. The infection rate was high (44%) among the residents of Padhrar. Molecular analysis (ITS1-RFLP) confirmed the presence of L. tropica, with ITS1 sequences exhibiting 100% similarity to isolates reported from India, Pakistan, Iran and Afghanistan. This study highlights the increasing number of CL cases in non-endemic areas, underscoring the importance of public health education, improved disease control programs, and cross-sector collaboration to mitigate the impact of CL and prevent its further spread.

Global access to clean and safe drinking water remains a formidable challenge, contributing to a myriad of health issues. This research exposes the existence of waterborne parasites in seemingly pristine spring waters, indicating potential contamination. Daily extensive sampling of Seventeen water sources was conducted in the untarnished freshwater streams of Tehsil Babuzai, District Swat, Khyber Pakhtunkhwa, Pakistan, from February to September 2021. Employing a stringent filtration process, the collected samples were effectively concentrated to detect any waterborne parasites. Subsequent application of the wet mount technique, combined with the capabilities of a compound microscope, revealed a disconcerting reality: all examined samples tested positive for various parasites. Identified parasites included Schistosoma species, Ascaris lumbricoides, Trichiuria trichiuria, Taenia saginata, Entamoeba histolytica, Amoeba, Lacrymana olor, Tintinnids, Paramecium, Dileptus, Euglena, Loxodes striatus, Acanthocyclops lynceus, Spondylosium, Oscillatoria, Cyanobacteria, Cilindros, Cilindros cerro, Commensal amoeba mature cysts,, Filliform larva of Strongyloides, Cercaria larva, Larva of Taenia solium, Egg of Enterobius vermiculais, Egg of Isospora belli, Egg of Tapeworm, Egg of Schistosoma species, Egg of Toxocara, and Egg of Diphyllobothrium latum. These findings clearly demonstrate the presence of a diverse array of parasites in the freshwater springs of Tehsil Babuzai, Swat, Pakistan. Implementing robust water treatment protocols, conducting regular monitoring and testing, and raising awareness about the risks of waterborne parasites are crucial steps to safeguard public health in the region.

Introduction: Thalassemia is a set of inherited disorder caused by a partial or complete mutation in one or more chain of the globin genes. Beta Thalassemia is the most common genetic disorder throughout the world including in Pakistan. Objectives: To assess the knowledge, attitudes and practices of parents of the children suffering from Thalassemia with regards to disease prevention. It also determined the degree of social and religious opposition faced by these parents during implementation of preventive practices Methods: A descriptive cross-sectional survey was conducted in district Swat, KP, via three regional thalassemia centers. Total of 200 parents of Thalassemia were interviewed using a pre-designed pre-tested structured questionnaire. All statistical analyses were conducted using Statistical Package for Social Sciences (IBM SPSS 23.0) Results: Out of the total participants, male parents were (67.7%), while females were (32.3%). A total of responses were gathered, and analyzed. Most respondents had adequate knowledge positive attitude towards thalassemia. Age, education, gender, and family income were key factors that correlated with or predicted the youth’s KAP towards thalassemia. Older respondents and women were more likely to have good KAP. 77.5% of parents opted for prenatal diagnosis, and 77% opted for termination of the affected fetus. Among these individuals, 7.5% faced opposition from their family members when they went for diagnostic testing, and this opposition rose to 18.5% when they had to opt for termination. The local religious clerics opposed prenatal diagnosis in 3.5% of the cases and termination in 4% of the cases. Conclusions: When compared to previous studies, the knowledge, attitudes, and practices of our study population were better. Thus, population screening programs have a positive impact on the knowledge, attitudes, and practices of careers of thalassemia patients. However, social and religious opposition needs to be countered for further implementation of effective preventive practices.

The core objective of forensic DNA typing is developing DNA profiles from biological evidence for personal identification. The present study was designed to check the validation of the IrisPlex system and the Prevalence of eye colour in the Pakhtoon population residing within the Malakand Division. Methods: Eye colour digital photographs and buccal swab samples of 893 individuals of different age groups were collected. Multiplexed SNaPshot single base extension chemistry was used, and the genotypic results were analysed. Snapshot data were used for eye colour prediction through the IrisPlex and FROG-kb tool. Results: The results of the present study found brown eye colour to be the most prevalent eye colour in comparison to intermediate and blue coloured. Overall, individuals with brown-coloured eyes possess CT (46.84%) and TT (53.16%) genotypes. Blue eye-coloured individuals are solely of the CC genotype, while individuals of intermediate eye colour carry CT (45.15%) and CC (53.85%) genotypes in rs12913832 SNP in the HERC2 gene. It was also revealed that brown-coloured eyes individuals were dominant among all age groups followed by intermediate and blue. Statistical analysis between particular variables and eye colour showed a significant p-value (SLC45A2 gene, rs12913832 SNP in HERC2 gene, rs1393350 SNP in SLC45A2, districts and gender. The rest of the SNPs were non-significant with eye colour, respectively. The rs12896399 SNP and SNP rs1800407 were found significant with rs16891982 SNP. The result also demonstrated that the study group differs from the world population based on eye colour. The two eye colour prediction results were compared, and it was discovered that IrisPlex and FROG-Kb had similar higher prediction ratios for Brown and Blue eye colour. Conclusions: The results of the current study revealed brown eye colour to be the most prevalent amongst members of the local population of Pakhtoon ethnicity in the Malakand Division of northern Pakistan. A set of contemporary human DNA samples with known phenotypes are used in this research to evaluate the custom panel’s prediction accuracy. With the aid of this forensic test, DNA typing can be supplemented with details about the appearance of the person from whom the sample was taken in cases involving missing persons, ancient human remains, and trace samples. This study may be helpful for future population genetics and forensics studies.

Fish are cold blooded vertebrates identification on the bases of morphology is not more precise and required high taxonomic expertise therefore molecular identification is used as an alternative and more accurate technique for the identification of fishes. In the current investigation 39 fish specimens were collected from May 2021 to February 2022. With this aim, the recent study was conducted in the freshwaters of River Swat in Malakand Division, Khyber Pakhtunkhwa, Pakistan to explore the genetic diversity of genus Triplophysa. DNA was extracted and amplified using gene specific primers. The PCR products carefully were sequenced and Phylogenic analysis was performed using neighbor-joining, maximum likelihood through MEGA software. Nucleotide composition and genetic pairwise distance recommend that the calculated species of the subfamily Nemacheilinae are organically dissimilar. The River Swat is the study area and it is inhabited by three Triplophysa species, Triplophysa naziri, Triplophysa microps, and Triplophysa choprai. The evolutionary tree shows that these species are clearly separated. The mean of total length calculated in the three species of the genus Triplophysa such as 8.175±0.198cm for Triplophysa naziri, 10.14±0.35cm for Triplophysa microps, 11.052±0.23cm for Triplophysa choprai. This study provides a model for the improvement of identification in cryptic diversity and field of ichthyofauna.

The present research was carried out to explore the spider fauna of Buner valley with taxonomic study from February 2018 to January 2019. For this purpose samples were collected, four times at each month from 4 tehsils: Daggar, Gagra, Mandan and Totalai. Two methods were used, hand picking and sweep net for collection of samples. During day and night, three habitats, arid area, agriculture land and building area were search for collection. A total of 534 samples of spider were collected from four sampling sites, in which 379 were belonging to family Araniedae. After confirmation, the identified species were belonging to 8 genera (Neoscona, Argiope, Cyclosa, Araneus, Cyrtophora, Larinia, Erivoxia and Poltys) and 19 species. 18 of them were identified to specie level while a single specie to its generic level. The genus Neoscona was the dominant genus 26.31% having 5 species while the genus Argiope 21.05% is the second dominant having 4 species followed by Cyclosa 15.78% having 3 species followed by Cyrtophora and Araneus 10.52% having two species both. The Poltys and Larinia 5.26% are the rarest genera represent single-single specie both. Statistical analysis show that specie richness (D) = 5.77, Simpson index (1-D) = 0.87, Shannon index (H) = 2.33. Diversity of spiders was evenly distributed and calculated Evenness value was H/InS = 0.5408. There is also few atypical species and Fisher alpha estimate high value (Fisher α) = 4.42. Chao-1 estimated we have reported 22 species.

Infectious agents cause serious diseases in humans worldwide and are responsible for the high rate of morbidity and mortality. The prevalence and epidemiology of infectious disease (HCV) in the hospital visited patients referred by the physicians through the initial findings and their associated risk factors were studied in Swat. The data of 174 infected patients were collected during the period of 2015 to 2017 from two clinical laboratories of Tehsil Matta Swat. Inform consent form was taken before blood collection. After taking informed consent blood samples were collected and ICT test was performed and then ICT positive cases were conform through PCR. A total of 174 ICT positive samples [106 male and 68 females] were included in this study. Age was considered from 10 to 72 years. Of the 174 ICT strip positive, 99 [63 males, 36 females] were confirmed through PCR. The prevalence rate was recorded 56.89%. I.V/I.M injection was recorded in 100% of the individuals. Visits to the barber shop was reported in (58%) of the individuals, married individuals were (81.0), surgical operation was reported in (44.8%), sharing toothbrush was observed in (29.9%), piercing was reported in (39.7%), family history was reported in (26.4%), dental treatment was observed in (21.8%), jaundice were (13.2%) and tattooing was (1.7%). Blood transfusion, surgical operations, Jaundice, family history and dental treatment were found significant risk factors for acquiring HCV infection. It was concluded that proper implementation of precautionary measures should be needed to control the spread of HCV in far near future.

For the proper growth of fish, it is necessary to feed the fish with a proper and balanced diet. A study was conducted to find out the effect of different protein-based diets on fingerlings of Tor putitora (mahseer). A feed with dietary protein levels of 35%, 40%, 45%, and 50% were prepared. The effect of different protein-based feed on weight gain, standard growth rate (SGR), food conversion ratio (FCR), percent weight gain, food conversion efficiency (FCE), and protein efficiency ratio (PER) was studied. An increase was observed in the growth rate with an increase in protein concentration up to 45%. The fingerlings fed a 45% protein diet shown the highest growth, followed by 50%, 40%, and 35%. The SGR value was greatest for 45% protein diet (8.56) followed by 50% and 40%, while the least values were observed for 35% protein feed (1.57). The same trend was observed for FCE. The highest PER values was observed in fishes fed 45% protein-based feed (0.65) followed by 50% (0.56), 40% (0.38) and35% (0.17). The food conversion ratio was lowest for 45% protein diet (3.41), while the greatest for 35% protein feed (16.85). It was concluded that a 45% protein-based diet was the best feed formulation for higher production of Tor putitora. However, research on the same percentage of protein diet is recommended for yearlings.

Plasmodium vivax is the most common human malaria parasite in Asian countries including Pakistan. Present study was designed to explore the genetic diversity of plasmodium vivax genotypes based on Pvmsp-3α and Pvmsp-3βgenes using allelic specific nested PCR and RFLP assays markers from field isolates in district Mardan, Pakistan. Blood samples of 200 P. vivax malarial patients were collected after taking their written informed consent. Genetic diversity in nested PCR products was determined by Restriction Fragment Length Polymorphism (RFLP) utilizing Alu1 and PstI restriction enzymes for alpha and beta gene products digestion, respectively. For analysis the genetic diversity of the sub allelic variants of Pvmsp3α and Pvmsp3β genes, Chi-Square test was performed by utilizing Minitab programming software 18. The P value 0.05 was considered as statistically significant. For Pvmsp-3α genes after gel electrophoresis of digested products, four distinct genotypes were obtained from total of 50 samples; type A: 35 (70%) (1.5-2.0 kb), 12 of type B (24%) (1.5-1.7 kb), 2 of type C (4%) (0.5-1.5) and one for type D (2%) (0.5-0.65 kb) which could be characterized into 9 allelic pattern (A1-A4, B1-B3, C1, D), in which A3 remained the most predominant. For Pvmsp-3βgenes, three distinct genotypes were obtained from 50 samples; 40(80%) of type A (1.5-2.5 kb), 9 (18%) of type B (1.0-1.5kb) and 1(2%) of type C (0.65 kb) which could be characterized into 6 allelic patterns (A1-A3, B1-B2, and C1). Most dominant one in Type A was A1 alleles which were noted (46%), while in Type B, the most dominant were B1 (10%).This study is the first ever report of molecular epidemiology and genetic variation in Pvmsp-3α and Pvmsp-3β genes of P. vivax isolates by using PCR/RFLP from District Mardan and showed a remarkable level of genetic diversity in the studied genes of circulating parasites in the study area. The results of this study will contribute in future studies about the genetic structure of parasite and vaccine development against the malaria.