Alcaligenes faecalis is one of the most important and clinically significant environmental pathogens, increasing in importance due to its isolation from soil and nosocomial environments. The Gram-negative soil bacterium is associated with skin endocarditis, bacteremia, dysentery, meningitis, endophthalmitis, urinary tract infections, and pneumonia in patients. With emerging antibiotic resistance in A. faecalis, it has become crucial to understand the origin of such resistance genes within this clinically significant environmental and gut bacterium. In this research, we studied the impact of antibiotic overuse in poultry and its effect on developing resistance in A. faecalis. We sampled soil and faecal materials from five poultry farms, performed whole genome sequencing & analysis and identified four strains of A. faecalis. Furthermore, we characterized the genes in the genomic islands of A. faecalis isolates. We found four multidrug-resistant A. faecalis strains that showed resistance against vancomycin (MIC >1000 μg/ml), ceftazidime (50 μg/ml), colistin (50 μg/ml) and ciprofloxacin (50 μg/ml). From whole genome comparative analysis, we found more than 180 resistance genes compared to the reference sequence. Parts of our assembled contigs were found to be similar to different bacteria which included pbp1A and pbp2 imparting resistance to amoxicillin originally a part of Helicobacter and Bordetella pertussis. We also found the Mycobacterial insertion element IS6110 in the genomic islands of all four genomes. This prominent insertion element can be transferred and induce resistance to other bacterial genomes. The results thus are crucial in understanding the transfer of resistance genes in the environment and can help in developing regimes for antibiotic use in the food and poultry industry.
Cytophaga hutchinsonii can efficiently degrade crystalline
cellulose, in which the cell surface cellulases secreted by the
type IX secretion system (T9SS) play important roles, but
the degradation mechanism remains unclear, and the anchor
mechanism of cellulases on the outer membrane in C.
hutchinsonii has not been studied. Here, chu_2177 was identified
by transposon mutagenesis and was proved to be indispensable
for cellulose utilization in C. hutchinsonii. Disruption
of chu_2177 resulted in O-antigen deficiency and chu_
177 could confer O-antigen ligase activity upon an Escherichia
coli waal mutant, indicating that chu_2177 encoded the Ontigen
ligase. Moreover, deletion of chu_2177 caused defects
in cellulose utilization, cell motility, biofilm formation, and
stress resistance. Further study showed that the endoglucanase
activity was markedly decreased in the outer membrane
but was increased in the culture fluid without chu_2177.
Western blot proved that endoglucanase CHU_1336 was not
located on the outer membrane but was released in the culture
fluid of the Δ2177 mutant. Further proteomics analysis
showed that many cargo proteins of T9SS were missing in
the outer membrane of the Δ2177 mutant. Our study revealed
that the deletion of chu_2177 affected the localization of
many T9SS cargo proteins including cellulases on the outer
membrane of C. hutchinsonii.
A novel Gram-stain-negative, aerobic, motile by means of
gliding, and short rod-shaped bacterium, designated strain
SH35T, was isolated from the dry surface of a tidal flat in
Hwasung-si, South Korea. Growth occurred at 10–40°C
(optimum 30°C), at pH 6.0–8.0 (optimum pH 7.0), in 1–12%
NaCl (optimum 2%), and was inhibited in the absence of
NaCl and Ca2+ ions. Phylogenetic analysis based on the 16S
rRNA gene sequences showed that strain SH35T belonged
to the genus Gramella and was a member of the family Flavobacteriaceae
with highest sequence similarity to Gramella
flava JLT2011T (96.1%), followed by Gramella oceani CCAMSZ-
TT (95.6%), and 93.0–94.9% to other recognized Gramella
species. The major cellular fatty acids (> 5% of the total)
of strain SH35T were iso-C15:0, Iso-C16:0, anteiso-C15:0, iso-C17:0
3-OH and summed feature 9 (C16:0 10-methyl and/or C17:1
iso ω9с). The major polar lipids were phosphatidylethanolamine,
two unidentified aminolipids and nine unidentified
polar lipids. The major respiratory quinone and the predominant
polyamine were menaquinone-6 (MK-6) and symhomospermidine,
respectively. The DNA G + C content was
40.5 mol% (39.7% based on total genome calculations). Based
on phylogenetic analysis and physiological and biochemical
characterization, strain SH35T represents a novel species of
the genus Gramella, for which the name Gramella fulva sp.
nov. is proposed. The type strain is SH35T (= KACC 19447T
= JCM 32369T).