Journal of Microbiology and Related Research

 Bluetongue disease (BT) is a vector borne infectious but noncontagious disease of wild and domestic ruminants. The BTV isolate (7bp) of sheep origin was inoculated to 9-11 day old chicken embryo followed by BHK-21 cell culture. Upon appearance of 75% cytopathic effect in cell culture viral nucleic acid was extracted. The viral nucleic acid showed BTV specific migration pattern of 3:3:3:1 in RNA-PAGE. The group specific ns1 gene RT-PCR confirmed the sample as BTV. The vp2 gene based serotype specific RT-PCR revealed the isolate as BTV16. The nucleic acid sequence of vp2 gene PCR products showed a high degree of identity (>99.0%) with other BTV16 isolates from different regions of India. It also showed maximum nucleotide identity of 99.7- 96.4% with several other eastern BTV16 viruses from India, Israel, Japan, Cyprus, Greece etc. Sequence identity study also revealed that 7bp isolate only showed 75.5% identity with western isolate of BTV16 from Nigeria. The phylogenetic study also showed a close relation between isolate in study and BTV16 isolates from India Japan, Israel and Greece which form a separate eastern cluster. Thus, molecular study showed that the isolate in study is of eastern origin and closer to BTV16 isolates from India, Greece, Japan, and Israel. 

Key words: Bluetongue Virus 16; Topotype; vp2 Gene; RT-PCR.

 Introduction: Urinary tract infection (UTI) in a very common bacterial infection in both sexes and all age groups and is also a major cause of hospitalization. E.coli is the most common causative organism in UTI with almost 85% isolates being E.coli . The main aim of antibiotic treatment is to completely eliminate the bacteria from the urinary tract. However, with the changing pattern of antibiotic sensitivity of urinary pathogens to commonly used antibiotics and the emergence of ESBL producing E.coli the situation has worsened. A new drug, Fosfomycin , has now been frequently used as an alternative drug in the treatment of uncomplicated UTI. The present study was carried out in our microbiology department with the aim of finding out the susceptibility pattern of Fosfomycin to frequently isolated urinary pathogens. Materials and Methods: This was a retrospective study and urine culture of patients from January to December 2015 were included in the study. A total of 2759 urine cultures of all ages and both sexes were included in the study. All urine specimens were inoculated on Blood and MacConkey agar plates and incubated at 370C for 24 hours .A growth of >105CFU/ml of bacteria was labelled as significant and was further processed.The bacteria were identified on the basis of Gram staining and colony morphology and accordingly the panel for identification & susceptibility was chosen to be processed on Vitek II ( (Biomerieux). Results: Out of 2759 urine cultures ,1341 (48.6%) cultures showed growth of bacteria. 1418 (51.3%) cultures were sterile .E. Coli (86.7%) was the commonest organism in followed by Klebsiella (8.50%), Pseudomonas (2.53%), Staphylococcus (1.41%),Enterococci (0.67%) and Proteus (0.14%). Highest sensitivity(100%) was observed in Proteus,followed by Pseudomonas(82.3%), Klebsiella(73.6%), Enterococci(66.6%) E.Coli(57.6%),and least for Staph.aureus(51.8%). Conclusion: The presence study suggests that resistance to Fosfomycin is on the rise and should be used with caution and resistance pattern should be monitored periodically. 

Keywords: Fosfomycin; UTI; Antibiotic Susceptibility.

 Introduction: Staphylococcus aureus is increasingly recognized as a cause of hospital associated (HA) and community associated (CA) infections. The Macrolide-lincosamide-StreptograminB (MLSB), family of antibiotics serves as one such alternative, clindamycin being the preferred agent due to its excellent pharmacokinetic properties. However widespread use of clindamycin led to increase in resistance due to target site modification mediated by erm genes which can be expressed either constitutively or inducibely so use of D-test in a routine laboratory enables us to guide clinicians in judicious use of clindamycin. Aims and Objective: To study prevalence of inducible and constitutive clindamycin resistance among Staphylococcus aureus and to compare in between MRSA and MSSA isolates. Material and Methods: A total of 107 Staphylococcus aureus isolates were subjected to routine antibiotic susceptibility testing including cefoxitin (30mcg) by Kirby Bauer disc diffusion method. Inducible clindamycin resistance was detected by using D test, as per CLSI guidelines on erythromycin resistant isolates. Results: A total of 67 isolates were resistant to erythromycin. Among 67 isolates, 17(25.37%) showed inducible Clindamycin resistance, 27(40.2%) showed MS phenotype and Constitutive resistance was seen in 23(34.3%) isolates. Constitutive and inducible clindamycin resistance was found to be higher in MRSA as compared to MSSA. Conclusion: For efficient use of clindamycin, D-test should be used as a mandatory method in routine disc diffusion testing to detect Inducible clindamycin resistance. 

Keywords: Constitutive; Inducible Clindamycin Resistance; MRSA; MSSA. 

 Ultrasonography machines are ideal vectors for cross infections. A busy machine may be used to scan many patients a day. The infection can be transmitted via ultrasound probes and coupling gel. Staphylococcus aureus, Enterococci, Klebsiella, Pseudomonas, E. coli are frequent cause of infections in both community and hospital. Organisms isolated from unclean US probe are important nosocomial pathogens and infections due to it are difficult to manage due to resistance to multiple antibiotics. So this study aimed to determine the percentage of bacteria isolated from unclean US probes and to determine the antibiotic sensitivity pattern.

Keywords: Ultrasonography; Bacteria; Antibiogram.

 Various microorganisms have been found to colonise the male genital tract which may play an important role in altering seminal parameters and thereby reducing male fertilizing potential. A total of 35 semen samples, obtained from PGIMER, Sector-12, Chandigarh, India were subjected to routine semen analysis according to WHO guidelines. The volume of all the samples, motility of 88.5% samples, pH of 80% of the samples was optimum. On culturing these samples on Brain Heart Infusion agar plates, it was found that 80% of the isolates so obtained were Gram positive cocci, 18% Gram negatives and 2% were yeast. Out of these isolates, 4% showed complete immobilization whereas 12% led to agglutination of spermatozoa. Scanning electron microscopy showed morphological alterations in sperm head, neck and mid piece etc. when incubated with the sperm immobilizing and sperm agglutinating strains. Further when the enzymatic activity was looked for, the results showed that these organisms were able to produce either protease, phospholipase, lipase or all of these. Hemolysis on sheep blood agar showed that only 2% of the isolates were capable of causing complete hemolysis and 6% showed partial hemolysis. The Gram positive bacteria were maximally sensitive to Oxacillin and Gram negative were sensitive to tobramycin and gentamycin. In conclusion, various Gram positive and Gram negative bacteria inhabiting male reproductive tract might produce certain factors which may impair sperm parameters. Eradication of these microbes by use of antibiotics can be a probable cure of microorganism induced infertility.

 Title : Blood stream Infections in Intensive Care Units. A Study from North India. Study Design: Prospective study. Place and Duration of Study: Sher-i-Kashmir Institute of Medical Sciences, Srinagar Kashmir. (July 2012 and Dec 2014). Methodology: A prospective analysis of blood specimens from various intensive care units (ICUs) was done over a period of four years. Antimicrobial susceptibility of culture positive isolates to various antibiotics was performed as per Clinical Laboratory Standards Institute (CLSI) guidelines.Gram-negative bacteria (GNB) were screened for extended spectrum -lactamase (ESBL) and metallo- ï¢-lactamase (MBL) production; whereas methicillin and vancomycin resistance was searched in staphylococci and nterococci isolates respectively. Results: The frequencies of Gram-positive and Gramnegative bacteria were 16.83% with yeast recovered in 5.78% of the specimens. Acinetobacter spp and K. pneumoniae were the most common Gram-negative bacteria and S. aureus the most common Gram-positive one. High level resistance to all the antimicrobials was seen; with Acinetobacter spp being the most multidrug resistant GNB isolated in the ICU setting. ESBL production was highest in K. pneumoniae isolates (77.1%). Also 49.6% of Acinetobacter isolates were found to be MBL producers. Methicillin resistance was seen in 95% of S. aureus and 91% of coagulase negative staphylococci (CoNS) isolates with vancomycin resistance seen in 46% of enterococcal isolates. Conclusion: An increasing trend over the years in the antibiotic resistance of blood stream pathogens in ICUs of this north Indian state was seen that calls for urgent measures to limit their continued rise.

Keywords: Gram-Negative Bacteria (GNB); Coagulase Negative Staphylococci (CoNS); Metallo-- actamase (MBL).

 Background and Objective: Health status of rural females is the most neglected in India. Hepatitis B is one of the highly infectious and sexually transmitted diseases which have a direct impact on the health of rural females as well on their siblings and family. So this study was undertaken to know the burden of Hepatitis B viral (HBV) infection in rural females so a prevention strategy can be made to curtail such dreaded infection in rural society. Method: A total number of 5035 female subjects of different age ranging from neonate to 90 years were grouped in 8 different groups designated as A to H. They were screened for detecting HBsAg both by Rapid Diagnostic tests and ELISA technique Results: Maximum number of subjects belonged to age group 21-30 yrs of group C, followed by group D(31-40yrs) and B(11-20 yrs). Of 5035 subjects, 256(5.08%) were positive for HBsAg. The positivity was maximum in both the extremes of life, 7.8%% & 21.9 t & 19.5% percent respectively. In other group the HBs Antigen positivity varied from 2.5 to 5.8 percent. Interpretation and Conclusion: The high incidence of Hepatitis B Viral infection clearly reflects the uneducated and neglected health status of rural females. Prevention strategy and education can reduce the further transmission and thereby reducing morbidity and mortality with HBV infection. 

Keyword: HBsAg; HBV; HBV in Females.