RESUMEN
Campylobacter concisus is an opportunistic bacterial pathogen linked with a range of human diseases. The objective of this study was to investigate the viable but nonculturable (VBNC) state of the bacterium. To induce the VBNC state, C. concisus cells were maintained in sterilized phosphate-buffered saline at 4°C for three weeks. The VBNC cells were monitored using quantitative analysis by propidium monoazide (PMAxx) coupled with quantitative real-time PCR (PMAxx-qPCR), targeting the DNA gyrase subunit B gene. The results demonstrated that C. concisus ATCC 51562 entered the VBNC state in 15 days, while ATCC 51561 entered the VBNC state in 9 days. The viable cell counts, assessed by PMAxx-qPCR, consistently remained close to the initial level of 107 CFU ml-1, indicating a substantial portion of the cell population had entered the VBNC state. Notably, morphological analysis revealed that the VBNC cells became coccoid and significantly smaller. The cells could be resuscitated through a temperature increase in the presence of a highly nutritious growth medium. In conclusion, under environmental stress, most C. concisus cells converted to the VBNC state. The VBNC state of C. concisus may be important for its environmental survival and spread, and the presence of VBNC forms should be considered in environmental and clinical monitoring.
RESUMEN
Campylobacter concisus is a bacterium that inhabits human oral cavities and is an emerging intestinal tract pathogen known to be a biofilm producer and one of the bacterial species found in dental plaque. In this study, biofilms of oral and intestinal C. concisus isolates were phenotypically characterized. The role of the luxS gene, which is linked to the regulation of biofilm formation in other pathogens, was assessed in relation to the pathogenic potential of this bacterium. Biofilm formation capacity was assessed using phenotypic assays. Oral strains were shown to be the highest producers. A luxS mutant was created by inserting a kanamycin cassette within the luxS gene of the highest biofilm-forming isolate. The loss of the polar flagellum was observed with scanning and transmission electron microscopy (SEM and TEM). Furthermore, the luxS mutant exhibited a significant reduction (p < 0.05) in biofilm formation, motility, and its expression of flaB, in addition to the capability to invade intestinal epithelial cells, compared to the parental strain. The study concluded that C. concisus oral isolates are significantly higher biofilm producers than the intestinal isolates and that LuxS plays a role in biofilm formation, invasion, and motility in this bacterium.
RESUMEN
The bacterium Helicobacter pylori is a human gastric pathogen that can cause a wide range of diseases, including chronic gastritis, peptic ulcer and gastric carcinoma. It is classified as a definitive (class I) human carcinogen by the International Agency for Research on Cancer. Flagella-mediated motility is essential for H. pylori to initiate colonization and for the development of infection in human beings. Glycosylation of the H. pylori flagellum with pseudaminic acid (Pse; 5,7-diacetamido-3,5,7,9-tetradeoxy-l-glycero-l-manno-nonulosonic acid) is essential for flagella assembly and function. The sixth step in the Pse biosynthesis pathway, activation of Pse by addition of a cytidine 5'-monophosphate (CMP) to generate CMP-Pse, is catalyzed by a metal-dependent enzyme pseudaminic acid biosynthesis protein F (PseF) using cytidine 5'-triphosphate (CTP) as a cofactor. No crystal-structural information for PseF is available. This study describes the first three-dimensional model of H. pylori PseF obtained using biocomputational tools. PseF harbors an α/ß-type hydrolase fold with a ß-hairpin (HP) dimerization domain. Comparison of PseF with other structural homologs allowed identification of crucial residues for substrate recognition and the catalytic mechanism. This structural information would pave the way to design novel therapeutics to combat bacterial infection.
RESUMEN
Salmonellosis, an acute invasive enteric infection, is endemic in Bangladesh. We analyzed 128,312 stool samples of diarrheal patients to identify Salmonella spp. during 2005-2013. A total of 2120 (1.7%) Salmonella spp. were isolated and the prevalence of Salmonella spp. decreased significantly over time (2â1%, P < 0.001). Among the typhoidal Salmonella (TS) serogroups, S. Typhi was predominant (404, [65.1%]) followed by S. Paratyphi B (139, [22.4%]) and S. Paratyphi A (78, [12.6%]). Of the nontyphoidal Salmonella (NTS) isolates, the serogroup C1 (560, [37%]) was predominant followed by B (379, [25%]), C2 (203, [14%]), E (127, [9%]), and D (94, [6%]). Most of the resistance was found towards nalidixic acid (40%), ampicillin (36%), cotrimoxazole (20%), chloramphenicol (13%), ciprofloxacin (4%), and ceftriaxone (4%). Interestingly, 32% of the isolates showed reduced susceptibility to Cip. Multiantibiotic resistance (MAR, ≥3 drugs) was more common among TS than NTS strains (P < 0.001). Among the representative ceftriaxone-resistant isolates, bla TEM gene was detected among 88% (7/8) of the strains, whereas only one strain of S. Typhi was positive for both bla TEM and bla CTX-M genes. The study reflects higher prevalence of MAR Salmonella spp. and is the first to report the bla TEM gene mediated ESBL production among Salmonellae in Bangladesh. Emergence of MAR Salmonella spp. in particular ESBL strains should be considered a public health concern.