Abstract
This study focus on cultivation, isolation and characterization of Bacteriocin from fresh cow milk (FCM) and fresh cow meat (FMS) samples obtained from Lapai Market in Niger State, Nigeria. Potential bacteriocinogenic bacteria were screened with agar diffusion method on culture plates seeded with Staphylococcus and E.coli . Bacteriocinogenic isolates from FCM were Enterococcus faecium FCM5 and Bacillus megaterium FCM6, while isolates from FMS include Bacillusbadius FMS1 and Micrococcus varians FMS2. Bacteriocins produced were assayed with pre-enriched MRS broth culture incubated at 30 0 C with adjusted pH (7.1) for 48 hrs and stored (4 0 C). The primed cultures were centrifuged (20,000rpm) for 30mins, supernatant were filtered (0.2μm membrane filters), precipitated with ammonium sulphate and steam sterilized (121 0 C at 15psi) for 15mins. Bacteriocin-like products (BLPs) were crystallized and tagged enterocin FCM5, bacillocin FCM6, bacillocin FMS1 and micrococin FMS2 according to bacteriocinogenic isolates' typing. After 48 hrs, Bacteriocinogenic load ranged between 5.1 - 6.13Log10 cfu/mL. Biopreservative indexes (BI) of BLPs (0.25 - 0.75 mg/mL) were effective against predominant food spoilers ( Saccharomyces , Pseudomonas , Klebsiella and Micrococcus spp isolated from range of beverages, starchy and meaty meals). Quantified BLPs (20 - 1880 IU/mL) strengths had no direct correlation with the antibacterial spectral (6 - 19mm) expressed as zone of inhibition. Thus, efficacy of BLPs observed in this study, were not a function of BLP quantity but quality. Conclusively,Enterocin FCM5, Bacilliocins (FCM6 and FMS1) and Micrococin FMS2 are thermal and pressure stable BLPs that are effective against predominant food spoilers. Keywords: Bacteriocin, cultivation, milk, meat, Lapai, bacteriociongenic
Highlights
Bacteriocins are proteinous metabolites of bacteria that have inhibitory activities against some related or non-related bacteria (Zhou et al, 2014)
This study aimed to cultivate, isolate and assess bacteriocin from fresh cow milk and meat samples obtained from Lapai Market in Niger State, Nigeria
1 mL of the solution was diluted serially with 0.1% peptone water upto the 3rd fold and 1mL of each sample were pour plated on lactic acid Bacteria medium (LABM), nutrient agar and De Man Rogosa sharpe (MRS) broth (Oxoid) incubated at 300C for 24 hours
Summary
Bacteriocins are proteinous metabolites of bacteria that have inhibitory activities against some related or non-related bacteria (Zhou et al, 2014). Bacteriocinlike products in combination with hydrogen peroxides, lactic acid and other organic acids are metabolites of LABs that naturally preserve foods from spoilage microbes. LABs that synthesize bacteriocins have been isolated from different food types and beverages such as beers, milk, fermented cucumber, cheese (Rekhef et al, 1995 Moham etal., 2012, Gonzales et al, 1994; Van Reenen et al, 1998) and fermented vegetables (Zhou et al.2014). LABs with bacteriocin-like metabolites have been isolated from meat products (Bromberg et al 2004). Bacteriocins (Nisin) status as GRAS (Mohammed et al, 2016), have increased detailed studies on the potency and application of other bacteriocin as natural alternative substance for food preservations. This study aimed to cultivate, isolate and assess bacteriocin from fresh cow milk and meat samples obtained from Lapai Market in Niger State, Nigeria
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