Comparison of microbiota and volatile organic compounds in milk from different sheep breeds.

In this study, we compared the microbiota and volatile organic compounds (VOC) present in the milk obtained from 3 different sheep breeds, namely Merino, Lacaune, and Assaf. Udder milk was collected from 21 animals, 7 from each breed. Bacterial microflora was determined metagenomically by extracting the DNA from the milk and analyzing the V3-V4 region of the 16S rRNA gene. Headspace solid-phase microextraction gas chromatography-mass spectrometry method was used to analyze VOC. The metagenomic analysis revealed (for Merino, Lacaune, and Assaf milk, respectively) Firmicutes (66.32, 69.36, and 57.08%), Actinobacteria (19.09, 7.67, and 19.40%), Proteobacteria (13.76, 21.06, and 22.19%), and Bacteroidetes (0.84, 1.91, and 1.33%) phyla in the milk samples. Lactobacillus was highly abundant in the milk of 3 breeds (29.64, 43.50, and 18.70%). The genera constituting more than 2% of all bacteria in all groups were Jeotgalicoccus (7.19, 5.34, and 10.77%), Enterococcus (5.18, 9.78, and 3.64%), and Corynebacterium (4.08, 3.00, and 13.44%). A total of 32 different VOC were identified by headspace solid-phase microextration analysis with 9, 30, and 24 different compounds from Merino, Lacaune, and Assaf breeds, respectively. Although ketone was the most abundant compound in Merino milk (71.84%), hydrocarbons were the most detected in Lacaune and Assaf milk (37.18% and 55.42%, respectively). A positive correlation was found between acetone, which was detected at the highest level in all groups, with Salinicoccus, Alloiococcus, Psychrobacter, and Dietzia. In addition, a negative correlation was found between the Lactobacillus genus, detected at the highest level in all groups, with methyl cyclopentane, 3-methylheptane, octane, decane, 3,3-dimethyloctane, and dodecane. Thus, differences were observed in the bacterial microflora and VOC in the sheep milk from different breeds under different feeding and breeding conditions.

Discrimination of viable and dead Vibrio parahaemolyticus subjected to low temperatures using Propidium Monoazide - Quantitative loop mediated isothermal amplification (PMA-qLAMP) and PMA-qPCR.

The aim of this study was to determine the effect of cold (4 °C) and subzero (-18 °C, -45 °C) temperatures on the occurrence time of membrane damage to provide Propidium Monoazide (PMA) penetration of Vibrio parahaemolyticus inoculated to the sea bass. Direct plate counting (DPC) and PMA-based quantitative loop-mediated isothermal amplification (qLAMP) and qPCR was utilized for discrimination of dead and live bacteria on the designated storage days (1, 3, 7, and 14). The optimum amount of PMA was 50 µM for inhibition of amplification derived from dead cells in spiked samples. The number of live V. parahaemolyticus was detectable at the end of the 14. day using PMA-qLAMP and PMA-qPCR at all the temperatures. On the 7th day, culturability has lost at any of the storage temperatures and DPCs at -18 °C and -45 °C revealed a difference of about 1 log10 CFU/ml between 1st and 3rd days. The same difference was also observed in PMA-qLAMP and PMA-qPCR on the same days (0.59-0.95 log10 CFU/ml). Subzero temperatures have the highest rate of viability while causing the fastest decrease in culturability in sample groups as a result of the higher level of transition to VBNC state. qLAMP and qPCR methods in the PMA-treated and nontreated groups on the storage days at all temperatures gave similar results (p > 0.05).