Friday, 25 July 2014

Chapter # 5 Discussion

Discussion:-
This body of work has largely been concerned with bacteriocin production from the E.coli. The bacteriocins are the proteinaceous substance act as the bactericidal agent that inhibit and often kill the some closely related species of the bacteria.  Bacteriocins inhibit the spoilage of food and also pathogenic bacteria.Bacteriocins are produced by both type of bacteria Gram –ve and Gram +ve bacteria. The bacteriocins from E.coli are of two type’scolicin which occupies the large molecular weight and microcin with low molecular weight. In this we only check that either bacteriocin produced from our isolated strains of E.coli or not.
We began this study by collecting the samples. The E.coli is belonging to the Enterobacteriaceae family it commonly found in gut flora of humans and animals. In the first step design the criteria how to collect sample we isolate pure cultures of E.coli by streaking method. Than by ager overlay method check the bacteriocin production but in the later studies the bacteriocin production was measured in the quantitative way but here our data is represent only which strain produced bacteriocin and which not. In our conducted study result indicates that 70% of E.coli strains produced bacteriocins.
In the next step we check the bacteriocin effect with different strains of S.aureus. Our results only explain the inhibitory effect on the S.aureus and have no quantitative data and shows that the S.aureus strains were inhibited by bacteriocins of E.coli but at very low level.The extracts from E.coli showed zones of inhibition against different pathogenic strains of diverse bacteria Staphylococcus aureus.
As in a study by Yang, et. al., a very little amount of bacteriocins was isolated due to adsorption on to the cell surface
The varied range of inhibition zones was observed but we cannot measure it. Sometime the bacteria acquired resistance against bacteriocins. such as in the study of Michael Feld garden and Margaret A. Riley in 1997 present  survey of colicin resistance in 158 E. coli isolates from a variety of hosts. They have screened for resistance in 137 isolates of non-E.coli enteric, which are close relatives of E. coli. Their data suggest that resistance levels are high. Similarly high levels of interspecific resistance to colicins are also observed. The frequency of colicin resistance to eighteen colicins is exceptionally high in all four E. coli collections surveyed. On average, 75% of the strains surveyed were resistant to a particular colicin.
Similarly Riley and Gordon in 1996 do the surveys of over 1000 isolates reveal that, on average, 30% of E. coli produce colicin.
In earlier study the David and Claire in 2005collect the 266 faecal isolates of Escherichia coli from humans was assayed for the production bacteriocins and screened using a PCR-based method for the presence of eleven colicins and seven microcins.
Our results are also in compliance of the previous work performed by Savadagoet al.(2006). However, in contrast, they have isolated eight strains of lactic acid bacteria from fermentedmilk to produce bacteriocins. Isolated bacteriocin showed inhibitory activity against Enterococcus faecalis, Bacillus cereus, Staphylococcus aureus and Escherichia coli. Gram-positive indicator bacteria were most inhibited. Their results indicate that the isolated strains of lactic acid bacteria are able to synthesize inhibitory substances against pathogenic bacteria. These inhibitory substances act differently on pathogenic reference indicator bacteria.
A recent study conducted by Adesokan et al. (2008) have observed less inhibitory activity against Gram negative bacteria as compared to Gram positive bacteria by studying the effect of bacteriocin on Staphylococcus aureus, Pseudomonas aeroginosa and Escherichia coli. It has been observed that the inhibiting substances produced by lactic acid bacteria are generally protein in nature (Klaenhammer, 1993).
In contrast to our work  Goraya, et.al in 2013 determine the Antibacterial activity of bacteriocins from LAB was evaluated against known food spoiling bacteria such as Escherichia coli, Enterococcus  faecalis and Staphylococcus aureus (obtained from Institute of  Microbiology UAF Faisalabad).A disc diffusion assay procedure was followed for it and measure the zone of inhibition.Most inhibited strain were Staphylococcus aureus and Enterococcus faecalis

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