RESUMO
Aims: The activation of cellular and humoral immunity depends upon nature of antigens. Complex proteins like bacterial outer membrane proteins (OMP) usually successfully activate both humoral and cellular immunity. Whereas antigens like bacterial lipopolysaccharides (LPS) usually elicit T-independent immunity i.e. humoral immunity without the activation of cellular immune wing. The present study was under taken to evaluate the comparative immunologic behavior of both the important molecules (bacterial lipopolysaccharide and outer membrane proteins) of Pasteurella multocida alone and in combination in bovine calves in field conditions. Methodology and results: Pasteurella multocida was isolated, purified and identified from an outbreak by mean of culture and biochemical methods. The pathogenicity of the confirmed isolates was done in rabbits (Oryctolagus cuniculus) on the principles of Koch’s postulates. Alum based vaccine against P. multocida was prepared and antibody titer against Outer membrane protein (OMP) and lipopolysaccharides (LPS) were determined by complement fixation test (CFT). The results showed that the antibody titer against OMP and LPS in whole culture vaccine is significantly higher than the respective tested vaccines. These results concluded that OMP no doubt is an active T-dependent immunogenic molecule but its immunogenicity increases many times when combined with LPS in whole culture vaccine. Conclusion, significance and impact of study: Lipopolysaccharides (LPS) in combination with outer membrane proteins (OMP) synergistically boost the humoral immune response in vaccinated animal.
RESUMO
Aims: The study was carried out firstly, to evaluate the prevalence of extended spectrum beta lactamase (ESBL), multidrug resistant Klebsiella pneumoniae isolates from Punjab, Pakistan and secondly, to characterize the genotypes of their beta lactamase producing enzymes and optimization of PCR based method for rapid and authentic detection of antibiotic resistant gene. Methodology and results: Two hundred of K. pneumonia strains were isolated from different clinical samples. Blood and MacConkey Agar were used to isolate and identify bacterial microorganisms while Muller Hinton Agar was used to evaluate the antimicrobial susceptibility against different antibiotics as per CLSI 2012 guidelines. ESBL producing bacteria were screened by double disk synergy /combination disk test. PCR was optimized and performed for resistant gene (CTX-M). The results showed that most of the isolates were resistant to multiple antibiotic including cephalosporin, aztreonam, sulphamethoxazole/trimethoprim, ciprofloxacin, doxycyclin and were sensitive to imipenam and amikacin. Frequency of CTX-M gene in ESBL producing K. pneumoniae was 94%. Conclusion, significance and impact of study: Based on the finding of this study it is suggested that prevalence of CTX-M gene (95%) is very high among ESBL producing isolates. Therefore, PCR based method may help clinicians for rapid detection and treatment of patients by choosing right medication against the resistant bacteria as early as possible.
RESUMO
Objective: To determine the residing microbial flora of ethylene oxide [EtO] sterilized medical devices and optimization of safe dose of gamma radiation [Cobalt 60 source] for the complete elimination of microbial load
Study Design: Experimental study
Place and Duration of Study: Department of Biotechnology, Lahore College for Women University, Lahore, Pakistan from September 2014 to June 2015
Methodology: Thirty-six samples of EtO sterilized medical devices of same batch of three different companies were collected for this study. Isolation and enumeration of microbes were done by using different selective and differential media. Gram staining and biochemically characterization by API 20 [Bio Merieux, France] kit was done for identification of the microorganisms. The medical devices having high microbial load were sent to Pakistan Radiation Services [PARAS] for gamma irradiations at 3 different selected doses [20 KGy, 25 KGy, and 30 KGy]
Results: Different types of Gram positive bacteria [Staphylococcus epidermidis, Staphylococcus aureus and Bacillus subtilis] were isolated from the EtO sterilized samples. Gram negative bacteria and fungi were not detected on these medical devices. Gamma irradiations results showed that 30 KGy was optimized dose for complete elimination of microbial flora on endotracheal, Nelaton, and tracheostomy tubes
Conclusion: Gamma radiations [Co 60 source] effectively decontaminate the microbial flora on the equipment previously sterilized by the ethylene oxide gas; and 30 KGy is the optimized dose for all these medical devices
RESUMO
To check the contribution of GLC3A locus to primary congenital glaucoma in the Pakistani population. We enrolled twenty-nine sporadic cases and three families with multiple individuals affected with recessive primary congenital glaucoma in the year 2013. It was a genetic linkage study accomplished jointly in Department of Biotechnology of Lahore College for Women University and School of Biological Sciences, University of the Punjab, Lahore. Samples from all affected individuals were checked for homozygosity for alleles of microsatellite markers spanning CYP1B1 at GLC3A locus. Genotyping was performed with fluorescently labeled primers by capillary electrophoresis. For familial cases, linkage was evaluated by checking the co-segregation of the phenotype with the genotypes. Two-point LOD score was calculated for each microsatellite marker with MLINK. Our study revealed that GLCA3 may contribute to glaucoma in 17% of the sporadic cases and patients in 2 of the 3 families. This data suggests that the GLC3A may make an important contribution to autosomal recessive primary congenital glaucoma in the Pakistani population. Genotyping and Sequencing of more families will be helpful to identify the common mutations in CYP1B1 in future