In the present study the

In the present study, the PCR assay used for the identification of F. verticillioides isolates was quick, accurate and more sensitive, as compared to conventional methods. It was shown that 194 out of 245 isolates were morphologically identified as F. verticillioides and this was further confirmed by VERTF1/VERT-R species-specific primers (Table 4). Among the remaining 51 isolates of Fusarium, 33 were identified as F. proliferatum and 18 as Fusarium species. F. proliferatum and F. verticillioides are having similar morphological characters. Fabrico Lanza et al. [41] did molecular identification of Fusarium isolates by PCR and species-specific primer pairs. Initially they identified five species as F. proliferatum based on morphological criteria but based on PCR test they were identified as F. verticillioides. Molecular analysis using species-specific PCR primers makes possible the precise identification of Fusarium species in such cases where morphological identification is not possible. PCR method also differentiates morphologically similar but toxigenically different F. verticillioides isolates. Present study revealed the association of 53.98% of fumonisin producing F. verticillioides with cereal samples collected from different regions of Karnataka state. In conclusion, data on the per cent incidence, frequency and relative density of F. verticillioides would be of great significance for predicting the extent of post-harvest infection, colonization and subsequent deterioration of cereals. Further, it also helps us to know the dry hippo pathway loss, nutritional changes and the extent of fumonisin levels during storage. Such a data is of immense value for assessing the possible health hazards in humans and animals upon consumption of such contaminated food grains. The high incidence of mycotoxigenic F. verticillioides is of primary concern for policy makers and food experts in this region to reduce the economic losses caused by these fungi and also to minimize the exposure of human and animal life to the potential risks of mycotoxins.
Conflict of interest
Acknowledgements We thank the Department of Science and Technology (DST-SERB) India, for providing grants through Young Scientist FAST TRAK research project (No. SR/FT/LS-176/2009; 30.04.2012) to Dr. M Y Sreenivasa, Principal Investigator. We thank the Institute of Excellence, University of Mysore, for their valuable support.
Introduction Many serotypes of Shiga toxin producing Escherichia coli (STEC) have emerged as a major cause of food-borne infections in the past 30 years [1–3]. E. coli serotype O157:H7 STEC is considered one of the most important pathogens for public health concerns, classified as an adulterant by the US Food and Drug Administration and the USDA Food Safety and Inspection Service (FSIS) [4,5]. Recently, it has also been recognized that a large number of E. coli non O157:H7 serotypes can be responsible for many E. coli outbreaks [6]. A study at the Centers for Disease Control and Prevention (CDC) showed that from 1983 to 2002 approximately 70% of non-O157 STEC infections were caused by strains from one of the six major serogroups known as “Big Six”, including O26, O111, O45, O121, O103 and O145 [7]. STEC can potentially cause a variety of clinical manifestations including diarrhea, hemorrhagic colitis and hemolytic uremic syndrome (HUS) [8]. Pathotypically, STEC can usually be classified as enterohemorrhagic E. coli (EHEC) [9]. As such, the FDA Bacteriological Analytical Manual (BAM) protocol now requires analysts who perform food testing to screen samples for both E. coli O157:H7 and non O157:H7 STEC [8,10]. For public health concerns, there is a need for rapid methods to identify, characterize and serotype pathogens associated with contaminated food during the surveillance and outbreak investigation. In addition to the traditional culture-based methods, several molecular assays including WGS, real-time (RT) PCR, DNA optical mapping. ELISA or whole genome microarray have emerged over the past several years [11,12]. Our study focuses on using RT-PCR analyses and DNA microarray methods to assist an outbreak investigation of a local E. coli non-O157:H7 outbreak causing diarrhea in a few people from a food borne outbreak.