Shafiq ur Rehman

Demographic and clinico-epidemiological features of dengue fever in Faisalabad, Pakistan.

This cross-sectional study was carried out to explore the epidemiological and clinical features of dengue fever in Faisalabad, Pakistan during 2011 and 2012. During the study period, anti-dengue IgM positive cases were reported in the post-monsoon period during the months of August–December. Certain hotspots for the dengue infection were identified in the city that coincide with the clusters of densely populated urban regions of the city. Out of total 299 IgM positive patients (male 218 and female 81); there were 239 dengue fever (DF) and 60 dengue hemorrhagic fever (DHF) patients. There was decrease in the median age of dengue patients from 31 years in 2011 to 21.5 years in 2012 (p<0.001). Abdominal pain was seen in 35% DHF patients followed by nausea in 28.3%, epistaxis in 25% and rash in 20% patients (p<0.05). Patients reported to be suffering from high-grade fever for an average of 8.83 days in DHF as compared to 5.82 days in DF before being hospitalized. Co-morbidities were found to be risk factor for the development of DHF in dengue patients. Clinical and laboratory features of dengue cases studied could be used for the early identification of patients at risk of severe dengue fever.

Screening of different contaminated environments for polyhydroxyalkanoates-producing bacterial strains

Total sixteen bacterial strains were isolated and purified from the samples collected from sugarcane molasses soil, sewage water and long-chain-hydrocarbon-contaminated area of the Punjab University, Lahore, Pakistan. Tolerance to different antibiotics was studied and strains showed multiple antibiotic resistance. All strains were characterized for Gram stain, biochemical reactions and polyhydroxyalkanoate (PHA) production. Total fourteen strains were Gram negative and two were Gram positive, while biochemically nine PHA producers showed affiliation to Pseudomonas, Enterobacter, Citrobacter, Bacillus and Escherichia. Screening for PHA production was done by Sudan black staining and nine out of sixteen strains exhibited PHA producing ability. PHA production was optimized for different growth parameters, like nitrogen concentration, pH and temperature. PHA extraction was done by solvent extraction method. Bacterial strains US1 and M1 accumulated up to 30% PHA of their cell dry weight on PHA extraction by solvent extraction method. Bacterial strain US1 was identified by 16S rRNA gene analysis as P. aeruginosa (DQ455691). PHA production was confirmed by PCR amplification of 500 bp fragment from PHA polymerase (Pha C) gene; five strains from nine PHA producers gave positive results on PCR. Pha C gene fragment of US1 was sequenced and submitted to Gene Bank under the accession number DQ455690. The amino acid sequence showed homology using the protein BLAST at 129–132 sites with different PHA synthases of the Pseudomonas sp.

Biodegradation of chlorpyrifos by bacterial genus Pseudomonas

Chlorpyrifos is an organophosphorus pesticide commonly used in agriculture. It is noxious to a variety of organisms that include living soil biota along with beneficial arthropods, fish, birds, humans, animals, and plants. Exposure to chlorpyrifos may cause detrimental effects as delayed seedling emergence, fruit deformities, and abnormal cell division. Contamination of chlorpyrifos has been found about 24u2009km from the site of its application. There are many physico‐chemical and biological approaches to remove organophosphorus pesticides from the ecosystem, among them most promising is biodegradation. The 3,5,6‐trichloro‐2‐pyridinol (TCP) and diethylthiophosphate (DETP) as primary products are made when chlorpyrifos is degraded by soil microorganisms which further break into nontoxic metabolites as CO2, H2O, and NH3. Pseudomonas is a diversified genus possessing a series of catabolic pathways and enzymes involved in pesticide degradation. Pseudomonas putida MAS‐1 is reported to be more efficient in chlorpyrifos degradation by a rate of 90% in 24u2009h among Pseudomonas genus. The current review analyzed the comparative potential of bacterial species in Pseudomonas genus for degradation of chlorpyrifos thus, expressing an ecofriendly approach for the treatment of environmental contaminants like pesticides.

The phage P.E1 isolated from hospital sewage reduces the growth of Escherichia coli

ABSTRACT This study involves partial characterisation of a lytic bacteriophage P.E1 against a multi drug-resistant clinical isolate of Escherichia coli, isolated from hospital sewage supply. The phage P.E1 has showed a narrow host range suitable for its use in phage therapy. Phage showed lytic activity up to 70°C and at alkaline conditions, but at higher acidic conditions its activity decreased. Latent period and burst size of P.E1 estimated from single-step growth curve was 40 min and 185 plaque-forming units per cell, respectively. The phage P.E1 reduced the growth of host bacteria during the initial 12u2005h of infection; however, the host bacteria developed resistance afterwards. During the 24-hour observation period, the bacteriophage could still reduce the growth of its host bacteria evident by lower optical density in the phage-treated samples compared with control. The phage genome was double-stranded DNA and larger than 12u2005kb in size. Further manipulations of genome and proteins may help to unveil the unique aspects of this phage, to use it in phage therapy against E. coli.

The Lytic SA Phage Demonstrate Bactericidal Activity against Mastitis Causing Staphylococcus aureus

Abstract Staphylococcus aureus is the major causative agent of mastitis among dairy animals as it causes intramammary gland infection. Due to antibiotic resistance and contamination of antibiotics in the milk of diseased animals; alternative therapeutic agents are required to cure mastitis. Lytic bacteriophages and their gene products can be potential therapeutic agents against bacteria as they are host specific and less harmful than antibiotics. In this study, Staphylococcus aureus were isolated from milk samples of the infected animals and identified biochemically. SA phage was isolated from sewage water showing lytic activity against Staphylococcus aureus isolates. The highest lytic activity of bacteriophages was observed at 37°C and pH 7, and the most suitable storage condition was at 4°C. SA phage efficiently reduced bacterial growth in the bacterial reduction assay. The characterization and bacterial growth reduction activity of the bacteriophages against Staphylococcus aureus signifies their underlying potential of phage therapy against mastitis.

Amelioration of Cadmium Stress in Gladiolus (Gladiolus grandiflora L.) by Application of Potassium and Silicon

ABSTRACT Gladiolus corms were grown in media contaminated with cadmium (Cd) (50 mg kg−1) and supplemented with silicon (Si) and potassium (K). The role of Si and K for mitigation of Cd toxicity was evaluated. Cd-induced stress generated significantly increased level of oxidative stress markers including hydrogen peroxide (H2O2), and malondialdehyde (MDA) in gladiolus. The application of K and Si improved the production of protein and proline in the treated plants. Moreover, K and Si supplemented plants exhibited an improvement in the activity of antioxidant enzymes and a reduction in the level of MDA, H2O2 and Cd uptake under Cd stress. Application of K and Si also enhanced the uptake of mineral nutrients including calcium (Ca), magnesium (Mg), manganese (Mn), sulfur (S) and K. The plants supplemented with K and Si exhibited a higher amount of total phenolics and flavonoids. The combined effect of Si and K was more pronounced regarding beneficial effects on gladiolus plants compared to individual effect of these elements under Cd stress. The current research reveals that Si and K may improve gladiolus growth by decreasing the oxidative stress and Cd uptake and by increasing the activity of antioxidant defense enzymes, the quantity of secondary metabolites and plant nutrition.

Assessment of biofilm removal capacity of a broad host range bacteriophage JHP against Pseudomonas aeruginosa

Pseudomonas aeruginosa is an efficient biofilm‐dwelling microbial pathogen, associated with nosocomial infections. These biofilm‐associated infections are resistant to antibiotics and immune defenses, therefore pose major problem against their treatment. This scenario demands alternative therapeutic regimens, and bacteriophage therapy is one among potential strategies for clinical management of multiple drug resistance. In this investigation, the efficacy of a bacteriophage, JHP, is evaluated to eradicate P. aeruginosa biofilms. Growth kinetics of P. aeruginosa biofilm revealed that the highest cell density biofilm (1.5 × 1016 CFU/mL) was established within the polystyrene microtiter plate at 72 h post inoculation. Pseudomonas aeruginosa biofilms of different ages, treated with JHP (0.6 MOI) for different post‐infection durations, reduced biomass from 2 to 4.5 logs (60–90%). JHP treatment before biofilm development reduced the bacterial load up to 9 logs (>95% bacterial load reduction) as compared with untreated control, which highlights its potential to prevent biofilm formation in indwelling medical devices. Combinations of JHP with other phages or antibiotics could be an efficient alternative for P. aeruginosa biofilm removal in clinical and industrial settings.

Isolation and characterization of lytic phages TSE1-3 against Enterobacter cloacae

Abstract The emergence of antibiotic resistant bacterial pathogens is becoming a major challenge for patient care. The utilization of alternative therapies for infectious diseases other than antibiotics is an urgent need of today medical practice. The utilization of lytic bacteriophages and their gene products as therapeutic agents against antibiotic resistant bacteria is one of the convincing alternative approaches. Here we present the isolation and characterization of three lytic bacteriophages TSE1-3 against Enterobacter cloacae from sewage effluent. The isolates maintained antibacterial activity for 10 hours of incubation followed by the development of phage resistance. Their stability at different temperatures and pH, established their possible application in phage therapy. The highest activity of the phages was observed at 37°C and pH 7.0, while they gave lytic activity up to 60°C. The latent period of all the TSE phages was 20 minutes, while the burst size was 360 for TSE1, 270 for TSE2 and 311 for TSE3. The phages were harboring double-stranded DNA larger than 12kb in size. Further research into the phages genome and proteins, animal experiments, delivery parameters and clinical trials may lead to their utilization in phage therapy.

Insight into Acinetobacter baumannii : pathogenesis, global resistance, mechanisms of resistance, treatment options, and alternative modalities

Acinetobacter baumannii, once considered a low-category pathogen, has emerged as an obstinate infectious agent. The scientific community is paying more attention to this pathogen due to its stubbornness to last resort antimicrobials, including carbapenems, colistin, and tigecycline, its high prevalence of infections in the hospital setting, and significantly increased rate of community-acquired infections by this organism over the past decade. It has given the fear of pre-antibiotic era to the world. To further enhance our understanding about this pathogen, in this review, we discuss its taxonomy, pathogenesis, current treatment options, global resistance rates, mechanisms of its resistance against various groups of antimicrobials, and future therapeutics.