Barkha Sharma

Occurrence of multidrug resistant Escherichia coli in groundwater of Brij region (Uttar Pradesh) and its public health implications

Aim: The study evaluates the microbial as well as physicochemical pollution of groundwater of Brij region of Uttar Pradesh, a major tourist destination in the country along with estimating the drug resistance evident in the isolated Escherichia coli. Materials and Methods: A total of 60 samples of groundwater were collected from six different sites and assessed for physicochemical (pH, color, taste, turbidity, total dissolved solids [TDS], total hardness [TH], chlorides, fluorides, nitrates, and iron) and microbiological parameters (standard plate count [SPC], most probable number test [MPN], E. coli). Results: A majority of the samples were found to be out of the range for most of the parameters except iron. Particularly, high values of TDS (up to 9000 ppm), TH (1500 mg/L), chlorides (3250 mg/L), fluorides (2.5 mg/L), and nitrates (100.2 mg/L) were observed at most of the sites in the region highlighting the fact that groundwater of the area is not potable. Samples were turbid and salty to taste. High SPC values, up to 3500 colony-forming unit/ml and coliforms beyond BIS range were found in 40% samples suggesting gross microbial contamination. Only 2 sites (G3 and G5) had low MPN values. Overall 16 (26.67%) E. coli were isolated with 3 (18.75%) producing red colonies on conge red agar, hence supposed to be pathogenic. No E. coli O157:H7 was isolated. High antimicrobial resistance was observed against amoxicillin and erythromycin, whereas E. coli isolates were sensitive toward cefotaxime-clavulanic acid and imipenem. 12 isolates (75%) were multidrug resistant (MDR) with MDR index >20%, and 2 isolates (12.5%) were found to be extended spectrum beta-lactamases positive. Conclusion: Groundwater is considered to be a safe option for potable water but it is obvious from the findings of this study that considerable physicochemical and microbial contamination is there in groundwater samples of Brij region. The occurrence of MDR E. coli in these waters is a matter of great public health concern.

A study on association of virulence determinants of verotoxic Escherichia coli isolated from cattle calves

Aim: The present study was conducted to find the association among virulence determinants of verotoxic Escherichia coli (VTEC) isolated from cattle calf feces. Materials and Methods: A total of 216 cattle calf fecal samples were collected aseptically and processed under required conditions for the isolation of E. coli. The isolates were further subjected to multiplex polymerase chain reaction (mPCR) for the detection of virulent genes. All the VTEC isolates were serotyped at the Central Research Institute, Kasauli, Himachal Pradesh. The VTEC isolates were observed for the enterohemolysin production on washed sheep blood agar (wSBA). Results: A total of 177 presumptive E. coli were isolated from 216 calf fecal samples revealing an overall prevalence of E. coli to be 81.94%. A total of 32 (14.81%) isolates were detected as VTEC through mPCR. The prevalence of verotoxin genes vt1, vt2, and combination of vt1+vt2 in the VTEC isolates was found to be 12 (37.5%), 14 (43.75%), and 6 (18.75%), respectively. Other virulent genes eaeA and hlyA were found in 6 and 11 VTEC strains with prevalence values of 18.75% and 34.37%, respectively. A total of 13 different O serogroups were revealed in serotyping of 32 VTEC isolates. Out of 32 VTEC strains, only 26 (81.25%) were enterohemolytic on wSBA as they produced the characteristic small, turbid zone of hemolysis around the streaking line. Although enterohemolysin production has been attributed to the presence of hlyA gene, only 11 of 26 enterohemolysin producing VTEC were found to be harboring the hlyA gene (11/26) 42.03%. Conclusion: The present study concludes that there might be an association between the presence of verotoxin genes and enterohemolysin production in VTEC group of E. coli.

Effect of incorporation of calcium lactate on physico-chemical, textural, and sensory properties of restructured buffalo meat loaves

Aim: The present study was conducted to develop a functional meat product by fortifying calcium (in the form of calcium lactate) with restructured buffalo meat loaf (RBML). Materials and Methods: Deboned buffalo meat obtained from the carcass of adult female buffalo within 5-6 h of slaughter and stored under frozen condition. Calcium fortified RBML were prepared by replacing the lean buffalo meat with calcium lactate powder at 0%, 1%, 1.25%, and 1.5% level through the pre-standardized procedure. The developed products were evaluated for physico-chemical properties, proximate composition, calcium concentration (mg/100 g), water activity (aw), Lovibond® tintometer color units, texture profile analysis (TPA), and sensory qualities as per-standard procedures. Results: Of the various product quality parameters evaluated, cooking yield (%), product pH, moisture (%), protein (%), fat (%), and water activity (aw) decreases significantly with increasing level of calcium lactate. Calcium content of fortified functional RBMLs was 135.02, 165.73, and 203.85 mg/100 g as compared to 6.48 mg/100 g in control. Most of the sensory scores at 1% and 1.25% levels of calcium lactate in treatment products remained comparable among themselves and control product, with a gradual decline. Conclusions: The present study concluded that 1.25% calcium lactate was the optimum level for the fortification of calcium in RBML without affecting the textural and sensory properties which could meet out 15% of recommended dietary allowance for calcium.

Genotypic Study of Verocytotoxic E. coli in Cattle by Multiplex Polymerase Chain Reaction

E. coli is the most commonly observed gastrointestinal flora of animals and environmental contaminant considered as important food borne pathogen causing serious complications in man and animals. The study was undertaken to detection of virulence gene using polymerase chain reaction (PCR) in cattle. In present study, a total of 160 samples were processed for isolation of verocytotoxic E. coli (VTEC). All samples were positive for Escherichia coli. Out of 160 Escherichia coli, 14 samples were found positive for VTEC. Out of 14 VTEC, 6 were found to be positive for stx1 gene (180 bp), 5 were stx2 (255 bp) and 3 were for stx1, stx2 and hlyA gene.