Page 90 - AIH-1-2

Artificial Intelligence in Health Combating XDR-bacteria as we approach 2050

local vendor near the Midnapore station area and chicken
meat from a local shop in Midnapore city. Antibiotic
plates containing ampicillin, cefotaxime, tetracycline,
streptomycin, ciprofloxacin, and erythromycin were
selected for the analysis of all collected samples.
Next, we presented a few biochemical data supporting
the heterogeneity of the MDR bacterial population in
chicken meat and human hair. Figure 7 illustrates the
results of the methyl red assay; Figure 8 demonstrates the
Figure 6. Detection of large plasmids and small plasmids in MDR
bacteria selected with six old drugs. Plasmid DNA was isolated from results of Simmon’s citrate utilization assay; Figure 9 depicts
6 mL bacterial overnight culture and loaded onto 0.8% agarose gel and the urea test results; and Figure 10 demonstrates the sugar
run for 4 h at 30 volts and ethidium bromide-stained (0.5 µg/mL in utilization test results to indicate the highly heterogeneous
1×TAE buffer). Lambda HindIII DNA and pBR322 plasmid DNA were population of MDR bacteria. We utilized E. coli KT-1_mdr
used as markers. Six bacteria were isolated from a chicken meat sample and P. aeruginosa DB-2_mdr as standards. The urea test
(Midnapore city), and six other bacteria were isolated from human hair
(salon in Kolkata city) and selected with six different antibiotic plates yielded negative results for both MDR bacterial strains,
while our selection of bacteria from the six antibiotic
plasmids and then transfected them into the Escherichia plates yielded few positive results. In addition, our
coli DH5α laboratory strain that had no mdr gene and was 16S rRNA gene sequencing identified Panalkaligenes and
highly sensitive to multiple antibiotics, either singly or in Stenotrophonomas bacteria (Figure 11). However, due to
combination. We observed ampicillin resistance in the 3 kb our laboratory’s lack of Biosafety Level 3 certification and
plasmid, whereas there was no chloramphenicol resistance the identification of potentially pathogenic MDR bacterial
in the 30 kb plasmid, suggesting that the cat gene was isolates in our data, we made the decision to suspend the
located in larger plasmids (>200 kb), making it challenging project.
to isolate using the conventional method from agarose Recent research by Williams et al. involved sequencing
gel at the Oriental Institute of Science and Technology the MDR bacterial population isolated from the stools of
(OIST) laboratory. Unfortunately, the plasmid contents workers in poultry farms in Bangladesh. They compared
in E. coli KT-1_mdr and E. coli KC-1_mdr were high and
heterogeneous, resulting in a smear instead of distinct the mdr genes with those found in poultry cecal and
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bands. Initially, we attributed this to contamination of our waste water resources. They discovered many mdr genes,
plasmid preparation with chromosomal DNA. Now, we including tetQ, blaTEM-1, blaSHV-1, and blaSHV-11,
understand that the MDR phenomenon is a mechanism in human fecal MDR bacteria. The other most abundant
of “genesis within” aimed at protecting bacteria in the mdr genes are macrolide-lincosamide-streptogramin-
intestine from the synthesis of 20 vitamins, which humans resistant genes. In the poultry cecal samples, however,
cannot produce on their own. Despite advancements, SHV-27, blaSHV-110, blaOXA-65, and blaOXA-641-like
hunger persists, and the world population still does not mdr genes were also located. In wastewater, the blaOXA-58
have access to a balanced diet every day. In addition, many mdr gene, associated with MDR Acinetobacter bacteria,
people, especially those from impoverished backgrounds, was found to be predominant, and such a protein
are reluctant to use multivitamin tablets. Consequently, could hydrolyze carbapenem drugs. The WHO has
we continue to depend on intestinal flora or probiotics. reported high contamination of poultry meat with CRE
All creatures, including humans, depend on nature, and Enterobacteriaceae (E. coli), Acinetobacter baumannii, and
all habitats on Earth live symbiotically in different ways. Pseudomonas aeruginosa with isolated mdr genes showing
Thus, it is important to conserve the environment and homology to previously described plasmid sequences
utilize phytoantibiotics, as proclaimed by the World Health (accession numbers: CP058135, LC75731, CP054855,
Organization (WHO). KY860573, and MN436715). 24,25

3.2. Isolation of chicken meat, milk, and human hair 3.3. Purification of abundant phytochemicals
bacteria and their characterization Our extraction and purification procedures for active
Our understanding suggests the presence of MDR bacteria chemicals from the ethanol extract using TLC are
in water, soil, and the intestinal tract. To investigate straightforward, as demonstrated in Figures 12 and 13.
further, we collected human hair from a salon in South We utilized eight 20×15 cm silica gel plates and processed
Kolkata, washed it with LB media, and plated it on 2.5 – 3 mL extract per TLC step in four tanks with lids.
LB-agar + ampicillin. Similarly, we obtained milk from a By employing repeated TLC, we were able to concentrate

Volume 1 Issue 2 (2024) 84 doi: 10.36922/aih.2284