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 AiM  Vol.7 No.5 , May 2017
Different Preservation Methods for Long Term Maintenance of Haemophilus influenzae
Abstract: Haemophilus species are Gram-negative coccobacilli that require factor X and factor V for growth. Beyond this, it is a finicky bacterium to culture, and any modification of culture procedures greatly reduces isolation rates. Poor quality of laboratories in developing countries results in its poor isolation rates. This study was done with the objective of finding out the optimal cultural environment and media so that it could be maintained for a longer period in economical settings like ours which was done using H. influenzae ATCC 49,766. In this study, several culture media were tested as a means to preserve H. influenzae ATCC like TSB + glycerol + sheep blood, BHI broth, BHI broth + glycerol, BHI broth+ glycerol + sheep blood, Chocolate agar slant and satellitism plate. Three sets of respective media were inoculated with 18 - 24 hours growth of H. influenzae. They were incubated at 37 oC 48 hours in a candle extinction jar. The media were checked for growth by subculturing them on chocolate agar plates and identified by biochemical reactions. Each set was maintained at 2 oC - 8 oC, -20 oC and at room temperature and checked for the viability 24 hourly by subculturing them on chocolate agar. Results showed best growth of H. influenza on chocolate agar slants for 15 - 20 days, followed by BHI + glycerol + sheep blood broth and satellitism plate for 4 - 6 days followed by BHI broth for 2 - 4 days. There was no growth in TSB + glycerol + sheep blood broth and BHI + glycerol broth media. Present study showed similar results as done by NS Srikanth et al. 2003 with growth on chocolate agar & satellitism plate for 3 - 5 days but no growth in TSB + Glycerol + Sheep blood broth media. Chocolate agar slant is by far the most long term preserving media for H. influenzae. However, growth on BHI broth with various modifications is also showed a good preservation for 3 - 5 days, so with further experiments we can hope to maintain the organism in these media also.
Cite this paper: Prajapati, K. , Rajdev, S. and Mullan, S. (2017) Different Preservation Methods for Long Term Maintenance of Haemophilus influenzae. Advances in Microbiology, 7, 343-348. doi: 10.4236/aim.2017.75028.
References

[1]   Musher, D.M. (1996) Haemophilus Species. In: Baron, S., Ed., Medical Microbiology. 4th Edition. University of Texas Medical Branch at Galveston, Galveston, Chapter 30.

[2]   Ananthanarayan, B.N. (2013) “Haemophilus” Textbook of Microbiology. 9th Edition, Universities Press, Hyderabad, 327-331.

[3]   https://www.cdc.gov/meningitis/lab-manual/chpt14-storage-shipping.html

[4]   Srikanth, N.S. and Macaden, R. (2003) Chocolate Glycerol Broth for Maintenance of Haemophilus influenzae. Indian Journal of Medical Microbiology, 21, 221.

[5]   de Saab, O.C.A., de Castillo, M.C., de Ruiz Holgado, A.P. and de Nader, O.M. (2001) A Comparative Study of Preservation and Storage of Haemophilus influenzae. Memórias do Instituto Oswaldo Cruz, 96, 583-586. https://doi.org/10.1590/S0074-02762001000400022

[6]   Clinical and Laboratory Standards Institute (2015) Performance Standards for Antimicrobial Susceptibility Testing; 21th Informational Supplement (M100-S25). Clinical and Laboratory Standards Institute, Wayne, 204-206.

 
 
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