Volume 8, issue 3

Methods: The current study was conducted to evaluate the prevalence of colibacillosis in young broiler chickens and antibiogram of E. coli. This study was done during February 2019 to June 2019; a total of 200 (n=200) liver samples of freshly dead young broiler chicks were randomly collected from the postmortem section of Veterinary research and disease investigation center Abbottabad (VRDICA). The identification of E. coli was confirmed by gram staining and biochemical tests. Susceptibility pattern to 13 antibiotics was also checked.

Results: Total of 200 (n=200) samples were brought to the VRDICA for the diagnosis of disease and antibiotic susceptibility testing. Out of these 130 were positive (Overall prevalence =65%).Area wise prevalence varied from area to area and highest prevalence was recorded in Abbottabad (81.11%) followed by Havelian (75%), Manghal (72.22%), Mansehra (60%), Qalandarabad (50%) and Haripur (28.57%). About age group, highest mortality was found in 11-15 days old chicks (93%) as compared to 6-10days (83.33%) and 1-5 days old chicks (21.42%). Antibiogram showed highest sensitivity to Gentamicin (100%) afterward Colistin Sulphate (92.30%). However, lincomycin (92%) and streptomycin (96%) display highest resistance.

Conclusion:  In this study, the highest prevalence of infection was observed in young chicks that play a crucial role in their mortality. E. coli had showed resistance to commonly recommended antibiotic so care should be taken while prescribing the drugs.  

Methods: Different rotten fruit samples were collected from different shops of Moon Market and Neelam block, Allama Iqbal Town, Lahore and used to purify bacterial cultures by growing on simple N-agar medium. Biochemical characterization was performed by different tests including gram staining, catalase, mannitol salt agar, glucose and fructose fermentation and nitrate reduction. Bacterial strains were further subjected to additional tests like HCN, H₂S production, metal resistance and antibiotic sensitivity.

Results: Nineteen bacterial strains were found positive for different tests and were characterized as Bacillus sp, Staphylococcus aureus, Micrococcus varians, Staphylococcus epidermidis and Staphylococcus saprophyticus. Many bacterial strains were resistant to antibiotics and high doses of most metals specially mercury. 

Conclusion: Due to the use of high doses of mercury for the storage of fruits, microorganisms have evolved resistance. It is an urgent need to take alternative measures for the storage of fruits for the safe lives.  

Methods: In the current study, remains of human bones are analyzed that dates back to 1000-1200 BC from different burial sites of Khyber Pakhtunkhwa province of Pakistan. Hypervariable region 1 (HVR1) as a genetic marker was selected. Multiple extractions, qubit assays quantification and cloning of PCR products were performed to get the sequences from the hypervariable region 1 of mtDNA. In this study, we also evaluate the possibility of extracting DNA from degraded human tissues (bones and teeth) such as remains buried in archaeological sites and remains of dead bodies buried for many decades. 

Results: Mitochondrial DNA HVR1 of 2000 years old degraded bone specimens were successfully determined by analyzing haplotype, defining polymorphisms in the hypervariable region 1. The prominent haplogroup found was eastern European R  (63%) that was also found in present day population by previous studies.

Conclusion: The protocol developed in this study could be used for the extraction of DNA from old degraded and ancient bone samples.  

Methods: For this, 11 samples of 5 different implants were taken from Tertiary Care Hospital Multan, Pakistan. Bacteria were isolated and identified by culture plate method. Tryptone soy broth (TBS) media was used for biofilm development by microbes in plastic tubes. Developed biofilm in tubes was visualized with crystal violet staining method and then biofilm forming potential was estimated by measuring the optical density through spectrophotometer. Antibiotic susceptibility was done by Kirby Bauer disk diffusion method to determine the resistance and susceptibility pattern of biofilm producers.

Results: Out of 11 different samples of indwelling medical devices, a total of 131 bacterial strains were isolated. The percentage of bacterial isolates which produced biofilms were Staphylococcus spp. (41%) followed by Escherichia coli (18%), Pseudomonas spp. (4%), Proteus spp. (7.2%), Klebsiella spp. (8.6%), Bacillus spp. (8.6%), Fusobacterium spp. (1.4%) Clostridium spp.(1.4%), Enterococcus spp. (7.2%) and Neisseria spp. (1.4%). Sixty-nine isolates were considered positive for biofilm formation while 58 were considered negative. The resistance was maximum against ampicillin (42%) followed by ceftazidime (17.1%), tetracycline (34%) and streptomycin (30%) while against vancomycin no resistance was observed.

Conclusion: Biofilms produced on medical implants by different bacteria are considered to be the major cause of hospital acquired infections and are very difficult to eradicate. These biofilms associated implant infections are challenging to treat because of their resistance towards various antibiotic therapies. Therefore, some efficient methods of prevention of biofilm formation should be introduced. 

Methods: A total of 150 unsexed broiler chickens were used for this experiment which were having five categories of treatment as T₁, T₂, T₃, T₄, and T₅. Each category had thirty birds. T₁ was treated with a standard diet, T₂was treated with acetic acid, T₃ with butyric acid, T₄ with citric acid and T₅ with formic acid.  The duration of this experiment was 28 days. After the specified time of this experiment, the data of uptake of feed and body weight was gathered on weekly basis.  Comparison of all the five treatments was done by using the Duncan's multiple range test.

Results: Feed Conversion Ratio (FCR) was found lower in T₅ than other treatments. T₅ group showed the highest average value of final body weight of broilers in contrast to the T₃ group which showed the lowest final body weight.  Feed intake was found significantly different within the treatments. T₃ showed significantly lower value as compared to other treatments. The lowest value of average regular uptake of grains was found in T₃ treatment group. Broilers fed on formic acid have shown a better protein efficiency ratio than that of butyric acid and citric acid. Broilers treated with citric acid have a significant difference which indicates more water consumption as compared to other treatments.

Conclusion: Organic acids have a productive effect on the growth of animals and broiler chickens. Organic acids including butyric acid, acetic, citric, formic, fumaric, and propionic acid vary in their biochemical actions in the system of animals. Organic acid affects the final weight gain, average regular gain in weight, total uptake of feed, and feed to gain ratio, daily intake of protein, protein efficiency ratio, total water uptake, average water intake, and water feed ratio. Based on the present study, further analysis is required to look over the impacts of addition of organic acid on the growth accomplishment of broiler chicks. 

Methods: The survey was conducted using a well-structured questionnaire that focused on the scope and potential of organic farming in Gilgit-Baltistan (GB), Pakistan. The data were collected from randomly selected 150 respondents in nine districts of GB between May and June (2019). This survey analyzed the factors influencing the willingness of farmers to pay attention to organic farming and evaluated the possibilities of espousal organic farming as an effective approach.

Results: In general, mixed perceptions were revealed by the behavior and attitudes of the farmers, however, it is necessary to notice the intentions of the farmers for substitute methods to make organic farming feasible. Based on the survey, people were not reluctant to adopt organic farming to ensure better market value for their products. As a result, they will sustainably get a better livelihood.

Conclusion: The current study is concluded that the espousal of organic farming could have a positive and noteworthy outcome on a farmer’s life in terms of profitability, compatibility, productivity, and sustainability in GB. There is a dire need for organic certification to increase the market value both at local and international levels. 

Methods: The current work focused on the exploration of probiotic potential of indigenously isolated Lactobacillus strains from dairy products and their encapsulation and utilization in yogurt formation.

Results: These Lactobacillus strains were identified as Lactobacillus fermentum MGA23-1 and Lactobacillus fermentum LMEM19 and found resistant to inhibitory substances like phenol (0.2%), bile salts (0.3%), pancreatin (0.5%) and pepsin (0.3%). The highest antibacterial activity was observed by Lactobacillus fermentum MGA23-1 against Pseudomonas aeruginosa (13mm). Encapsulation experiment showed that the number of bacterial CFU/g increased significantly (p < 0.05) in beads during storage up to 7 days. Chemical characterization of microcapsules was assessed using FTIR and showed characteristics wavelength major at 1541 – 1716 cm^-1 and 3336 cm^-1. Yogurt was prepared using a single probiotic strain, in a consortium of Lactobacillus fermentum MGA23-1 and Lactobacillus fermentum LMEM19 and in the form of beads. Best results were observed in the case of microencapsulation.

Conclusion: It was concluded that both strains had the potential to be used as a probiotic in the dairy industry. 

Method: Blood samples were collected in EDTA coated vacutainer from patients after their consent. DNA was extracted from serum through the phenol-chloroform method; Hepatitis B surface gene was cloned in TA cloning vector, subclone in pET 28a expression vector. An expression vector containing the Surface gene was then transformed into a competent bacterial host BL21 and inducted with IPTG at 0.1-0.2mM concentration for expression. The expressed proteins (soluble and pellet form) were analyzed on SDS PAGE.

Results: Hepatitis B Surface gene of 681bp after PCR were detected under UV light then successfully cloned and subcloned in pET 28 expression vector. The restricted fragment indicating the gene of interest was 681bp when analyzed on 1.2% Agarose gel under UV light. The required protein of 25kDa was obtained in soluble form when detected on 12% SDS PAGE after staining with Coomassie Blue dye.

Conclusion: Hepatitis B surface gene was successfully expressed in both insoluble and pellet forms using E.coli. The expression of surface protein needs to maximize through optimizing conditions to be used as potent candidate for vaccine production to prevent hepatitis B infection.  

Methods: A cross sectional online study was conducted. A self-developed online questionnaire was made by Google forms and sent to the author’s network with local residents in Pakistan. The questionnaire contained 35 questions among which 6 questions were related with the demographic features, 14 questions assess knowledge of participants, 4 questions evaluate the attitude, 7 questions assess the practices of the participants towards COVID-19 and 4 questions assess the psychological impact towards COVID-19.

Results: Among the participants (n=528) 66.7% were female and 33.3% were male. Among them 49.8% of the participants were from the age group of 18-24 and the percentage of students was higher i.e. 39.8%. The overall range for correct answers for the knowledge questionnaire was 97.5%-46.8%, for attitude 75.2%-91.9% and for practices 39%-95%. 79% of the participants were confident that Pakistan will overcome the disaster situation of COVID-19.

Conclusion: The findings of the current study indicated that Knowledge among the Pakistani residents about COVID-19 was found satisfactory. A significant number of the participants still lacking confidence when compared with the other countries. However, to effectively control the infection spread and improving COVID-19 knowledge the well-structured awareness programs must be launched by the government those could be helpful for Pakistani residents to hold optimistic attitudes and maintain appropriate practices. 

Methods: In this study, we analysed DNA methylation levels of 48 tumour suppressor genes loci in 24 bladder tissues (19 bladder cancer samples and 5 control samples taken from histologically normal bladders).  DNA Methylation analysis was done using Human Tumour Suppressor Genes EpiTect Methyl II Complete PCR Array from Qiagen ^TM.

Results: We identified significant difference in DNA hypermethylation levels at APC, BRCA1, CDH1, CDH13, CDKN2A, DAPK1, ESR1, FHIT, MGMT, RASSF1, SOCS1, TIMP3, TP73, VHL, WIF1 between controls and cancerous samples. It was also observed that CADM1 and DKK3 were differentially methylated in non-muscle invasive versus muscle invasive bladder cancer samples. Additionally, DNA hypermethylation of ESR1 was notified as the novel tumour suppressor gene specific for the Saudi population in bladder cancer.

Conclusion: Our findings suggest that these aberrant DNA methylation patterns in bladder cancer are disease and population specific and have a potential to develop as distinct DNA methylation-based biomarkers in future. 

Methods: Immunoinformatics approach was used to design subunit vaccine (SV) by joining 8 MHC-I bindings, 6 MHC-II bindings, and 8 B-Cell epitopes with AAV, GPGPG, and KK amino acid linkers, respectively. The efficacy of the SV was enhanced through Mtb protein Rv3763 (LpqH, PDB ID= 4ZJM) as an adjuvant at the N-terminal of SV. The in silico analyses evaluated the SV to predict allergenicity, antigenicity, and physico-chemical properties.

Results: Predictions revealed that SV is non-allergic and highly antigenic. The physico-chemical analysis showed that the SV was stable and basic in nature. The three-dimensional structure of SV was stable with a high binding affinity against the mouse TLR2 receptor. In silico cloning suggested the effective transformation of SV into the eukaryotic expression vector.

Conclusion: This study permits preclinical validation of the designed SV in mouse host to confirm its immunogenic potential and efficacy, which will help in controlling tuberculosis.