QUANTIFICATION OF Staphylococcus aureus AND Escherichia coli FROM BOVINE SUBCLINICAL MILK SAMPLES BY CONVENTIONAL PCR
Journal Title: Journal of Experimental Biology and Agricultural Sciences - Year 2018, Vol 6, Issue 5
Abstract
The present study explores the application of conventional PCR and Mole v/s Avogadro’s number using single copy genes of Staphylococcus aureus and Escherichia coli for assessing bacterial load in milk samples from mastitis cases. The nuc and uidA genes produced the lowest detection limit for Staph. aureus and E. coli respectively. The present findings estimated that, as few as 60 and 89 genome copies or organisms of Staph. aureus and E. coli were detected, respectively. Based on the LOD of molecules, standard graphs for these genes were generated and this knowledge was applied to milk samples from the field. Spiking known amount of genomic DNA (which in turn indicates organisms based on Avogadro’s number) in LB broth and pasteurized milk was carried out to compare the input with output and simulating field conditions. A total of 90 samples from subclinical cases of mastitis were collected from four organized farms located at various villages. Out of 90 samples, 29 (32.22 %) were showed culture and duplex PCR positive. Of these, 12 (13.33 %) were found to be positive for E. coli alone, nine (10 %) were found to be positive for Staph. aureus alone, and eight (8.88 %) carried a mixed infection for both. In conclusion, this study suggests that quantification of bacteria directly from milk by the conventional PCR can be an alternative to time consuming conventional culture method and expensive real-time PCR, but requires extensive standardization.
Authors and Affiliations
K. M. Chandrashekar, Shrikrishna Isloor, D. Rathnamma, B. M. Veeregowda, Raveendra Hegde, R. Sharada, G. S. Naveen Kumar, Nagendra R. Hegde
Keywords
Related Articles
- EP ID EP593081
- DOI 10.18006/2018.6(5).808.815
- Views 60
- Downloads 0
An overview on the functional food concept: prospectives and applied researches in probiotics, prebiotics and synbiotics
The principal role of diet is to supply adequate nutrients providing energy to sustain physiologic functions and well-being. Every foods and feeds are functional and consumption of bioactive molecules is facilitated by d...
Nutritional quality of maize in response to drought stress during grain-filling stages in mediterranean climate condition
Maize is considered one of the most essential dietary components in human food and animal feeding. The objectives of the present study were to quantify the effects of drought stress on qualitative traits of maize at grai...
EVALUATION OF FORMOL GEL SLIDE TEST WITH REFERENCE TO BLOOD MICROSCOPY AND ENZYME LINKED IMMUNOSORBANT ASSAY FOR DIAGNOSIS OF SURRA IN CATTLE
The objective of the present study was to evaluate the conventional Formol Gel Slide Test (FGST) vis-a-vis Enzyme Linked Immunosorbant Assay (ELISA) in diagnosis of Trypanosoma evansi infection in cattle. In all, 51 adul...
AEROBIC VERSUS ANAEROBIC GERMINATION PERFORMANCE OF SELECTED RICE (Oryza sativa L.) GENOTYPES WITH OR WITHOUT SUBMERGENCE TOLERANCE
Ten selected rice genotypes adapted to rainfed lowland ecosystem of Assam was evaluated for their ability to germinate and grow underwater (anoxic condition) in comparison to the aerobic condition. Anaerobic germination...
MICROBIAL POPULATION WITH POTENTIAL TO SIMULTANEOUSLY DEGRADE ORGANOCHLORINE INSECTICIDES
Organochlorine pesticides (OCPs) such as 1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane (p,p’-DDT) and ?-hexachlorocyclohexane (Lindane) pose serious health effects to the environment and community. Although both these c...