Isolation and Identification of Lactic Acid Bacteria from Buffalo’s Raw Milk in Basrah Province by Sequencing the 16S rRNA.

Abstract

Milk is a nutrient substance that screted from mammary glands of mammals. Lactic acid bacteria (LAB)
of raw milk are natural microflora and might be starter cultures for milk fermentation. This study aimed to
isolate LABs from raw buffalo’s milk and identify them by 16S rRNA sequencing for possible uses in the
manufacture of food product., A total of 30 buffalo’s raw milk samples were collected from different areas
of Basrah province (Shut-Alarab,Al-Zubair,Al-Qurna, Basrah conter and Abi-Elkhasib through the period
extended from October to December 2019 to isolate the LABs using conventional bacteriology test and
identify them by 16S rRNA sequencing using Polymerase Chain Reaction assay (DNA-based method).
DNA extraction was carried out in duplicate for all raw milk samples. The results showed buffalo’s milk
raw showed positive result for bacteriological test (Gram positive-catalase negative)and for the presence
of the 16S rRNA gene. There was no significant difference among ages, months, and area of the current
study. Buffalo’s raw milk at >4-8 year of age showed a higher ratio of LABs (71.43%) compared with that
of buffalo at >1-4 year of age (47.83%). In addition, the highest ratio of LABs in buffalo’s raw milk was in
October (66.6%) and the lowest one was in November (36.3%). Furthermore, the highest ratio of LABs in
buffalo’s raw milk was in Abi- Elkhasib (100 %) and the lowest one was in AL-Zubair (20%). In conclusion
Lactic acid bacteria were found in buffalo’s raw milk. Biochemical and genotypic tests are effectively used
to identify the LABs. All LABs isolates could be survived well in buffalo’s raw milk in any age per year,
months, and area of study. These findings suggest the possibility that LABs isolates from buffalo’s raw milk
might use to inhibit pathogenic and spoilage bacteria in the food products and might improve flavour and
quality of the food products.