The spore forming Bacillus cereus (66) was screened for the induction of β-lactamase in presence of an inducer using iodometric assay. A significant induction in marker enzyme was observed in B. cereus 66 at maximum residual limit (MRL) of penicillin, ampicillin, cloxacillin, amoxicillin, cefalexin, and cephazolin belonging to β-lactam group of antibiotics. A microbial based assay, where enzyme induction was optimized at pH 7.0, temperature 30°C, and whey powder (0.25%) after 4 h of incubation. The spore based assay was tested with milk samples spiked with 6 different β-lactam antibiotics. The results were 100 and 83.33% in correlation with microbial receptor and inhibition based assay, respectively. Overall, spore based assay can be a useful and cost effective tool for the specific detection of β-lactam group of antibiotics in milk.
Calf diarrhoea remains the biggest challenge both in the small and large farms. Infectious diarrhoea is associated with many pathogens, E coli being one but, majority are systematically treated with antibiotics. Since antimicrobial resistance (AMR) is a growing menace, the need to find alternative prophylactic solutions using popular kitchen herbs such as Trachyspermum ammi (Carom seeds), Curcuma Longa (Turmeric) and Cinnamon (Cinnamomum sp.) extracts is been investigated against virulent form of E. coli isolated from calf diarrhoea. The virulence factors identified in these isolates were ST (32.5%), LT (20%), eaeA (15%), stx1 (2.5%) and stx2 (5%) with the occurrence of the most common serogroups as O18 (15%) followed by O111 (12.5%). Highest resistance was seen with β lactamase inhibitor (Amoxicillin/Clavalinic acid) followed by β lactams (Ampicillin, Cefuroxime and Cefepime). Antibacterial effect of cinnamon (methanol) and carom seed (ethanol) extracts was found to be highly sensitive at higher concentration of the extract (500 to 250 µg/ml) while, minimum inhibitory concentration (MIC) of turmeric was significantly higher (P<0.05) followed by cinnamon. Turmeric, cinnamon and carom had the potency of inhibiting the pathogenic E coli which maybe suggestive of its use in calf diets as prophylaxis against diarrhoea.
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