Microbiological analysis of hospital cafeteria fast foods in preference to the presence of antibiotic resistant bacteria and their association with pathogenicity

Abstract

Antibiotic resistance is an emerging national and global threat to the food chain. Rising popularity of fast foods along with their capacity of harboring different multiple antibiotic resistant pathogenic bacteria displays a leading issue related to public health across the globe. The research focused on the identification of various multidrug-resistant (MDR) bacterial species that are currently in circulation, along with the existence of virulence and antibiotic resistance determinants among the isolates of eight hospital cafeteria fast foods (sandwich and burger), as well as the evaluation of the level of knowledge and related practices involved with food handling amidst the food handling personnel of these hospital cafeterias in Dhaka, Bangladesh. The total viable bacterial counts from burger and sandwich samples ranges from 2.1× 102 to 4.0× 106 (CFU/g) and 7.0× 102 to 1.0 × 106 (CFU/g) respectively. According to the total bacterial count, the majority of the sandwich and burger samples were in an unsatisfactory condition. A total of 137 bacterial species were isolated from the sandwich and burger samples, of which 87 (63.50%) were Gram-negative and 50 (36.50%) were Gram-positive. Among the Gram-negative isolates, 52.87% were retrieved from sandwiches, while 47.12% were from burgers. In case of Gram-positive isolates, 58% and 42% were retrieved from sandwiches and burgers respectively. The phenotypic characteristics of the isolates were screened through selective identification (morphological) in different selective media and Gram staining followed by different biochemical tests and gene specific PCR of ribosomal gene (16S rRNA) sequencing. DNA sequence homology and phylogeny analysis were employed to find out the close relatives. The Gram-negative isolates were distributed into 10 different genotypes using Randomly Amplified Polymorphic DNA (RAPD) typing namely Escherichia spp., Enterobacter spp., Klebsiella spp., Salmonella spp., Pseudomonas spp., Citrobacter spp. and Acinetobacter spp. The Gram-negative isolates were widely dispersed among the various bacterial species, with Klebsiella spp. accounting for the highest percentage (42.53%) and Salmonella spp. and Acinetobacter spp. the lowest (1.15%). Based on various biochemical tests, the isolated Gram-positive organisms were divided into 6 groups encompassing Bacillus spp., Planococcus spp., Micrococus spp., Streptococcus spp., Clostridium spp. and Staphylococcus spp., with the highest percentage of isolates from Bacillus spp. (58%). and lowest in Staphylococcus spp. (2%). About 16.09% of the Gram-negative and 6% i of the Gram-positive isolates were strong biofilm formers, while 13.79% and 6% of the Gram-negative and Gram-positive strains showed β-hemolytic properties respectively. Using 17 antibiotics from 11 different antibiotic classes, the susceptibility pattern against antibiotics was examined. Approximately 80.99% of the Gram-negative and 54% of the Gram-positive isolates were resistant against multiple antibiotics. The study isolates had significantly higher percentages of resistance to particular antibiotic classes; with the maximum resistance against ampicillin and penicillin preceded by meropenem and the lowest resistance against imipenem. The virulence and antibiotic resistance genes among the Gram-negative isolates were detected by employing gene specific PCR. Among the harbored virulence genes, all Pseudomonas spp. were oprI positive and 12.5% were oprL positive. Salmonella isolate was invA positive and 50% of the E. coli strains were uidA positive. In case of antibiotic resistance genes, blaSHV showed the highest prevalence (20%) among the class A β-lactamase; amidst the carbapenemases resistance, blaOXA-48 showed the highest prevalence (12.22%) followed by blaOXA-1 (2.22%), blaKPC-1 (2.22%). Through the fluoroquinolone resistance, gyrB showed the highest prevalence (50%) followed by qnrS (41.67%). Most of the food handlers had poor knowledge (62.50%) and practice scores (71.67%). The training condition (P<0.05), educational status (P<0.05) and monthly income (P<0.05) significantly influenced the practices of the food handling personnel as revealed by univariate analysis. According to the outcomes of multivariate logistic regression, food handling personnel practice score was significantly influenced by the level of education (P<0.05) and training status (P<0.05, AOR=0.049, 95% CI =0.011, 0.225). Food handler’s knowledge and practice score was significantly related (P<0.05) to each other. Prevalence of such virulence and resistant determinants along with poor food handling knowledge and practices in hospital cafeteria fast food indicates the risky situation regarding hospital acquired pathogen, increases the risks for already infected and vulnerable patients, impedes treatment and threatens recovery as well as increases the health costs of the country and may jeopardizes the attainment of sustainable development goals (SDGs) particularly the SDG 3.