MOLECULAR AND BIOCHEMICAL CHARACTERIZATION OF PSEUDOMONAS SPECIES ISOLATED FROM SUBCLINICAL MASTITIS MILK AND ICE CREAM AND ITS SUSCEPTIBILITY TO ALLIUM SATIVUM AND COMMIPHORRA MOLMOL PLANT EXTRACTS IN EGYPT.

Mastitis is the most important economic illness affecting dairy cattle worldwide.Milk is nutrient and essential daily diet for growing human and young animals. Also, serve as good medium for several microbial growth. Identification of contamination milk pathogens still depends principally on culture and phonotype method. This study aimed to isolate, characterize and evaluate Pseudomonas spp. isolated from subclinical mastitis raw milk and ice cream by using molecular and biochemical assays manual and automated system in Egypt. Sixty (60) random milk samples were collected aseptically from dairy farms (cow and buffaloes), supermarkets and street venders. Also, Fifty (50) Ice cream samples were collected from supermarkets and street venders at four different Governorates in Egypt. Number of positive pathogenic bacteria were isolated from raw milk (26) and (22) from Ice cream samples. Not only Pseudomonas SPP were isolated from samples but also major pathogenic bacteria were isolated as E.coli, Staphylococcus auraus and Salmonella. Putative Pseudomonas spp were partial characterized by biochemical test, API 20NE, Enzymatic activity lipase, lecthinase and proteolytic activity, automated system (VETIC system). Molecular characterization by polymerase chain reaction (PCR through detection of aprA and exotoxin A genes, amplifying a 396- pb region of the exotoxin A and 1580-bp region of apr A genes. The inhibitory values of essential oil were evaluated by disc diffusion test and minimal inhibition concentration (MIC) for each oil plant extracts compared with reference antibiotics. Commiphorra molmol essential oil extract has a marked inhibitory effects against pseudomonas spp ( 26.6 ± 0.33 ) followed by A. sativam essential oil extract IZD were (21.6±0.48) respectively. P.aeruginosa was as the most predominant isolate followed by P. fluoresces. Diagnosis of Pseudomonas spp with molecular methods is a rapid, sensitive and accurate method for identification, detection of virulence factors and differentiation of Pseudomonas spp isolated from dairy milk and ice cream. .Also, essential oils from Commiphorra molmol and Allium sativam have a marked antibacterial effect against Pseudomonas spp and may be use a novel antibacterial agents and food preservative but it’s still need extensive study for safety and clinical study in vivo.