Introduction: Bacterial isolates and control strains stocking is an integral part of clinical microbiology laboratories. This is an essential step in maintaining quality. Preserving the strains without altering the character is an essentiality. There are different stock culture preparations studied in past showing varied level of performance. Aim: To evaluate the performance in terms of longevity and phenotypic character preservation of Peptone Glycerol Broth (PGB) in comparison to Brucella Glycerol Broth (BGB) and Skim Milk (SKM). Materials and Methods: The present study was a prospective analytical study. Three quality control strains and seven clinical isolates with different types of resistance were stocked in triplicates with cryobead based peptone broth with 15% glycerol, Brucella Broth (BB) with 15% glycerol and 10% SKM and stored at -80°C. Isolates were revived in monthly pattern, quarterly pattern and once after 10 months to assess the variations in viability and loss of phenotypic properties arising out of repeated freeze thaw and contaminations. Viability was assessed by time taken to produce observable confluent growth on revival. Metabolic characters and antibiotic susceptibility testing were compared before and after stock revival at intervals. Results: The phenotypic characters like metabolic features and antibiotic susceptibility were preserved with all three preparations both with repeated freeze thaw and single freeze thaw at the end of 10 months. PGB and BGB had a 100% revival rate of stored isolates with a confluent growth at 24 hours in comparison to 93.56% with SKM. Conclusion: Cryobead preparation of peptone broth-15% glycerol can be used as an effective preparation for stock culture maintenance of non-fastidious bacteria and yeast.
Sepsis, the second leading cause of death is due to infections. Intensive care units (ICUs) are having the highest burden of treating the patients with sepsis and nosocomial infections compared to other areas of hospitals. Our objective was to identify the bacteriological profile and their antibiogram of sepsis cases in all ICUs. A sum of 102 blood samples were collected from patients with clinically suspected sepsis with elevated CRP. Processed by an automated method using Bact/Alert & growth were identified by Standard guidelines. Out of 102 samples, 54 (53%) were shown positive by culture. Gram-negative bacilli were the predominant and their number were 33 (61.1% ) and the commonest organisms were from the Enterobacteriaceae family. Escherichia coli was the highest number with 15 (27.7%) followed by Klebsiella pneumoniae 10 (18.51%), & the rest were single isolates of Salmonella typhi, Proteus mirabilis and Citrobacter koseri. Nonfermenter isolated were Acinetobacter baumanii 3 (5.6%), Pseudomonas aeruginosa 2 (3.7%). The Gram-positive cocci were 17 & 32.4% of culture positivity. Coagulase-negative Staphylococcus was the highest isolated accounting for 9 (16.6%) followed by Staphylococcus aureus 6 (11.1%) and Enterococcus faecalis (3.7%). Culture positivity will be more when CRP is also included in the selection of samples for sepsis and Gram-negative bacilli are the leading cause in septicemia and organisms belonging to the Enterobacteriaceae family still dominate in septicemia infections in ICUs and a real challenge for treatment are MDRs which needs to be detected regularly by using screening tests.
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