Abstract
This study was carried out in order to isolate and diagnosis of common bacteria from bovine mastitis in Baghdad city which have the ability to produce biofilm and detect its molecular composition. Twenty five milk samples were collected from different regions in Baghdad city from udders of cows suffering from clinical and subclinical mastitis.Then cultured on blood agar. Gram staining was done to differentiate between bacteria gram positive which cultured on Mannitol Salt Agar and Nutrient Agar while gram negative bacteria were cultured on MacConkey agar and Eosin Methylene Blue agar. All bacterial isolates were subjected to different biochemical tests, API 20 E System, API Staph System and RapIDTMONE System kit to confirm the diagnosis.Christensen tube method was used to detect the ability of the diagnostic bacterial isolates to produce biofilm. Specific forward and reverse primers were designed according to a program from NCBI-Genbank for Staphylococcus aureus (Genbank code: gb |KR265472.1|), for Escherichia coli : gb|JQ781567.1| and for Klebsiella pneumoniae gb|KT944736.1|, To study the sequence of these genes after amplification of 16srRNA genes by using PCR was appeared because it is sensitive and highly specific assay serve as suitable molecular diagnostic tool for detection and compare these genes sequencing with references strains.Results showed that 8 (32%) out of 25 milk samples were positive for Staphylococcus , 6 (24%) out of 25 samples were positive for Klebsiella pneumonia . and 11(44%) were positive for E.coli . The results showed that 22 (88%) isolates out of 25 milk have the ability to produce biofilm . Genetic identities results showed that Staphylococcus aureus and Klebsiella pneumonia isolates gave 99% matching and resembling the reference strains while Escherichia coli isolates identity showed resemble match of 100% with the reference strain
Highlights
Mastitis is inflammation of the parenchyma of the mammary gland regardless of the cause, mastitis characterized by a range of physical and chemical changes in the milk and pathological changes in the glandular tissue[22]. the most important changes in the milk include discoloration, the presence of clots and the presence of large numbers of leukocytes, there is swelling, heat, pain, and odema in the mammary gland in many clinical cases [19]
The search for a successful treatment of biofilm infections that can prevent and eradicate biofilms in the clinical environment is still ongoing [21].From all major pathogens in bovine mastitis biofilm behavior is best studied in Staphylococcus aureus, the reason for the preference of S. aureus is presumably related to the importance of this species both for bovine mastitis and human [17]
There are a variety of ways in which biofilms can cause health issues in humans,as well as animals with wounds and mastitis being two common clinical challenges in animals, research is still compiling evidence on the role of biofilms in veterinary medicine,much of the information regarding biofilm infection and disease has been extrapolated from human research and applied to the veterinary environment as there are still many gaps in veterinary biofilm research [9].The diseases caused by a particular strain of E. coli depend on distribution and expression of many virulence determinants such as adhesion
Summary
Mastitis is inflammation of the parenchyma of the mammary gland regardless of the cause, mastitis characterized by a range of physical and chemical changes in the milk and pathological changes in the glandular tissue[22]. the most important changes in the milk include discoloration, the presence of clots and the presence of large numbers of leukocytes, there is swelling, heat, pain, and odema in the mammary gland in many clinical cases [19]. The most important changes in the milk include discoloration, the presence of clots and the presence of large numbers of leukocytes, there is swelling, heat, pain, and odema in the mammary gland in many clinical cases [19]. The formation of bacterial biofilms of E.coli and Klebsiella pneumonia in a host in general seems to be, based on current evidence, to a large extent anintracellularevent[4]. There are a variety of ways in which biofilms can cause health issues in humans ,as well as animals with wounds and mastitis being two common clinical challenges in animals, research is still compiling evidence on the role of biofilms in veterinary medicine ,much of the information regarding biofilm infection and disease has been extrapolated from human research and applied to the veterinary environment as there are still many gaps in veterinary biofilm research [9].The diseases caused by a particular strain of E. coli depend on distribution and expression of many virulence determinants such as adhesion
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