Study of antagonistic effects of Lactobacillus strains as probiotics on multi drug resistant (MDR) bacteria isolated from urinary tract infections (UTIs)

Atiyeh Naderi, Roha Kasra-Kermanshahi, Sara Gharavi, Abbas Ali Imani Fooladi, Meghdad Abdollahpour Alitappeh, Parvaneh Saffarian, Atiyeh Naderi, Roha Kasra-Kermanshahi, Sara Gharavi, Abbas Ali Imani Fooladi, Meghdad Abdollahpour Alitappeh, Parvaneh Saffarian

Abstract

Objective(s): Urinary tract infection (UTI) caused by bacteria is one of the most frequent infections in human population. Inappropriate use of antibiotics, often leads to appearance of drug resistance in bacteria. However, use of probiotic bacteria has been suggested as a partial replacement. This study was aimed to assess the antagonistic effects of Lactobacillus standard strains against bacteria isolated from UTI infections.

Materials and methods: Among 600 samples; those with ≥10,000 cfu/ml were selected as UTI positive samples. Enterococcus sp., Klebsiella pneumoniae, Enterobacter sp., and Escherichia coli were found the most prevalent UTI causative agents. All isolates were screened for multi drug resistance and subjected to the antimicrobial effects of three Lactobacillus strains by using microplate technique and the MICs amounts were determined. In order to verify the origin of antibiotic resistance of isolates, plasmid curing using ethidium bromide and acridine orange was carried out.

Results: No antagonistic activity in Lactobacilli suspension was detected against test on Enterococcus and Enterobacter strains and K. pneumoniae, which were resistant to most antibiotics. However, an inhibitory effect was observed for E. coli which were resistant to 8-9 antibiotics. In addition, L. casei was determined to be the most effective probiotic. RESULTS from replica plating suggested one of the plasmids could be related to the gene responsible for ampicillin resistance.

Conclusion: Treatment of E. coli with probiotic suspension was not effective on inhibition of the plasmid carrying hypothetical ampicillin resistant gene. Moreover, the plasmid profiles obtained from probiotic-treated isolates were identical to untreated isolates.

Keywords: Antibiotic resistance; Lactobacillus; Pathogenic bacteria; Probiotic; Urinary tract infection.

Figures

Figure 1
Figure 1
Frequency of all bacterial strains isolated from urine samples
Figure 2
Figure 2
Frequency of antibiotic resistance in isolated Escherichia coli from urine samples
Figure 3
Figure 3
Frequency of antibiotic resistance in isolated Klebsiella pneumoniae from urine samples
Figure 4
Figure 4
Frequency of antibiotic resistance in Enterobacter strains isolated from urine samples
Figure 5
Figure 5
Frequency of antibiotic resistance in Enterococcus strains isolated from urine samples
Figure 6
Figure 6
Isolation of ampicillin sensitive mutant strains created by treating the strain number 5 with Ethidium bromide and acridine orange using replica plating method. Plate A: plate without ampicillin; plate B: plate containing ampicillin
Figure 7
Figure 7
Isolation of ciprofloxacin sensitive mutant strains created by treating the strain number 5 with Ethidium bromide and acridine orange using replica plating method. Plate A: plate without ciprofloxacin; plate B: plate containing ciprofloxacin
Figure 8
Figure 8
Agarose gel electrophoresis showing plasmid patterns of Lane 1: small colonies growth on plates containing amikacin after treating with ethidium bromide; Lane 2: maternal strain
Figure 9
Figure 9
Agarose gel electrophoresis. Lane 1: plasmid patterns of the strain number 5; Lane 2: plasmid patterns of the strain number 5 after treatment with ethidium bromide; Lane 3: plasmid patterns of the strain number 5 after treatment with acridine orange; Lane 4: maternal strain
Figure 10
Figure 10
Agarose gel electrophoresis showing plasmid pattern of strain number 6. Lane 1: treated strain with L. casei supernatant; Lane 2: treated strain with L. acidophilus supernatant; Lane 3: treated strain with L. rhamnosus supernatant

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