Pakistan Journal of Biochemistry and Biotechnology

Characterization of Pectobacterium Carotovoratum Subsp. Carotovorum causing Bacterial Soft Rot Disease of Turnip in Pakistan

Salman Ghuffar — 2023-06-21

This study aims to identify the Pectobacterium carotovorum subsp. carotovorum (Pcc) causing bacterial soft rot disease of turnip in Pakistan through conventional and molecular techniques. During the survey, four different bacterial isolates (VRSPCC01-04) were obtained from symptomatic turnip steckling (cv. purple top) at Vegetable Research Station Sahiwal, Punjab, Pakistan, and subjected for further analysis. All the isolates were characterized and identified based on morphological, biochemical, and molecular analysis (16S Ribosomal RNA). During morphological studies, the colony characters found deep cavities or pits formation in Crystal violet pectate medium (CVP), Creamy yellowish colony color was found on Yeast Extract Glucose calcium carbonate Medium (YGC), while creamy in color with the oval-shaped colony was observed on nutrient agar (NA) medium. In addition, the Pathogenicity test proved that all the tested isolates developed a typical water-soak symptom after 24 h and initiation of soft rot began the 48 h after inoculation on turnip roots. Results of biochemical tests showed that all isolates had a positive reaction for catalase, citrate, and an adverse reaction for Phosphatase and Indole test. The 16S rRNA sequences of isolates were 99-100% similar to the corresponding 16S rRNA sequence of the Pectobacterium carotovorum (PCC) reference isolates. Based on this, all identified bacterial isolates are first time reported as a causative agent of soft rot disease of turnip in Pakistan.

Knowledge, Attitudes and Practices Regarding the Prevention of Thalassemia in Parents of Thalassemic Children in Swat

Murad Ali Rahat — 2023-06-21

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.

CRISPR/Cas9-mediated Inhibition of Vacuolar Invertase (VInv) Gene in Potato

Tahmina Sattar — 2023-06-30

Potato is the third most important food crop worldwide after the wheat and rice. It has potential to overcome the issues of world’s food security. Potato crop faces some challenges during storage as Cold Induce Sweetening (CIS). During cold storage, the accumulation of reducing sugars such as fructose and glucose takes place in potato tubers. These reducing sugars later react with free amino acid and produce dark-brown pigmentation on potato products during frying. That leads towards reduction of potato quality and consumption. The increased quantity of sucrose is further hydrolyzed by Vacuolar Invertase (VInv) gene into reducing sugars. VInv gene acts as a key player in inducing CIS in potato. The objective of study was cloning of CRISPR/Cas9 construct and successful inhibition of VInv gene function in potato. Agrobacterium-mediated transformation of VInv gene in potato tissues was performed. The knockout of VInv gene in transformed plants was analyzed by using specific primers through polymerase chain reaction (PCR). Real time polymerase chain reaction (RT-PCR) was carried out to analyze the expression of transgene in transgenic potato plants. The knockout of VInv gene using CRISPR/Cas9 strategy could maintain the quality of potato tubers during cold storage.

A Review and Analysis of Heavy Metals in Freshwater Reservoirs of Pakistan: their Bioaccumulation, Biological Magnification and Biotic Transmission

Bushra Nasir Khan — 2023-06-30

Problems associated with pollution are becoming more severe as industrialization advances rapidly. Inorganic and organic wastes, both water-soluble and insoluble, are discharged into the aquatic environment, contributing to one of the most critical problems: water pollution. The discharge of waste in water channels allows heavy metal ions, which are toxic to all kinds of life, to enter the food chain. Lead, nickel, sodium, potassium, zinc, copper, iron lithium, cadmium, and chromium (VI) are among the most hazardous metal ions. Because they don't break down, metal ions build up in the environment and move up the food chain. Therefore, higher trophic level animals are more susceptible to their toxicityHeavy metals found in freshwater and sediments have been found to bioaccumulate in fish tissues. Rawal Dam, Mangla Dam, and Zabi Dam were identified as highly contaminated areas of freshwater that had detrimental effects on fish and human health. The Rawal Dam, which received a lot of industrial and sewage drainage from Punjab's metropolitan areas and industry, was significantly contaminated compared to other Pakistani Dams. Changhoz and Ghol Dams, with their massive water flows and relatively few accompanying industrial facilities, have the healthiest ecosystems of all Pakistan's dams. Changhoz Dam's freshwater fish are considered to be safe for human consumption. Heavy metals are accumulated by migratory birds at Rawal Dam from eating fish from freshwater reservoirs. Heavy metal pollution has been linked to a variety of social, ecological, and economic issues. The detrimental effects of heavy metals on aquatic organisms and human well-being can be reduced if wastewater is purified before being released into freshwater.

Virtual Screening for the identification of potential inhibitors for Vp35 protein of Marburg Virus

Hafsah Muhammad — 2023-06-30

Marburg virus (MARV) causes Marburg hemorrhagic disease (MHD) or Marburg hemorrhagic fever (MHF) in humans as well as in Non-Human Primates with a high mortality rate. It is a category A, risk group 4, an extremely dangerous pathogen that can also be used as a bioweapon. To date, no licensed medication against MARV infection is available. The
current study attempts to design putative antiviral drugs or vaccine candidates against vp35, a polymerase cofactor protein of Marburg virus, using a computational virtual screening approach. From the zinc database 1600 compounds were selected for docking purpose. Top 20 molecules obtained via virtual screening approach were considered as potential inhibitors against the polymerase cofactor VP35 of MARV, based on Lipinski’s rule of five, binding affinity between -6.1 to -5.6 kcal/mol, a Root Mean Square Deviation (RMSD) value of zero, strong hydrogen bonding with vp35 and proper Absorption, Distribution, Metabolism and Excretion (ADME), hence can inhibit VP35. The features of these drugs can be further analyzed on the basis of in-vivo and in-vitro analysis in order to recommend any of these drugs for trials in humans and NHPs against Marburg hemorrhagic fever.