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Microbiology Spectrum Sep 2023The first cases of in Argentina were detected in three (Pre) recovered from two hospitals in Buenos Aires city in 2013. The isolates were genetically related, but the...
The first cases of in Argentina were detected in three (Pre) recovered from two hospitals in Buenos Aires city in 2013. The isolates were genetically related, but the plasmid profile was different. Here, we characterized the -harboring plasmids of the first three cases detected in Argentina. Hybrid assembly obtained from short- and long-read sequencing rendered in Col3M plasmids of . 320 kb (p15268A_320) in isolate PreM15268, 210 kb (p15758B_210) in PreM15758, and 225 kb (p15973A_225) in PreM15973. In addition, PreM15758 harbored a 98-kb circular plasmid (p15758C_98) flanked by a putative recombination site (-Tn), with 100% nucleotide ID and coverage with p15628A_320. Analysis of PFGE/S1-nuclease gel, Southern hybridization with probe, hybrid assembly of short and long reads suggests that pM15758C_98 can integrate by homologous recombination. The three -plasmids were non-conjugative . Moreover, genes were incomplete, and oriT was not found in the three -plasmids. In two isolates, bla was embedded in a partially conserved structure flanked by two IS. In addition, all plasmids harbored , , and D1 genes and , , B3, and as part of a class 1 integron. Also, p15268A_320 and p15973A_225 harbored . To the best of our knowledge, this is the first report of clinical harboring bla in an atypical genetic environment and located in unusual chimeric Col3M plasmids. The study and continuous surveillance of these pathogens are crucial to tracking the evolution of these resistant plasmids and finding solutions to tackle their dissemination. IMPORTANCE Infections caused by carbapenem hydrolyzing enzymes like NDM (New Delhi metallo-beta-lactamase) represent a serious problem worldwide because they restrict available treatment options and increase morbidity and mortality, and treatment failure prolongs hospital stays. The first three cases of NDM in Argentina were caused by genetically related recovered in two hospitals. In this work, we studied the genetic structure of the plasmids encoding in those index cases and revealed the enormous plasticity of these genetic elements. In particular, we found a small plasmid that was also found inserted in the larger plasmids by homologous recombination as a co-integrate element. We also found that the plasmids were not able to transfer or move to other hosts, suggesting their role as reservoir elements for the acquisition of resistance genes. It is necessary to unravel the dissemination strategies and the evolution of these resistant plasmids to find solutions to tackle their spread.
PubMed: 37732774
DOI: 10.1128/spectrum.01651-23 -
Journal of Travel Medicine 2014
Topics: Diarrhea; Escherichia coli; Female; Humans; Irritable Bowel Syndrome; Male; Providencia; Travel
PubMed: 24734916
DOI: 10.1111/jtm.12111 -
Applied and Environmental Microbiology Jan 2018Flies have the capacity to transfer pathogens between different environments, acting as one of the most important vectors of human diseases worldwide. In this study, we...
Flies have the capacity to transfer pathogens between different environments, acting as one of the most important vectors of human diseases worldwide. In this study, we trapped flies on a university campus and tested them for mobile resistance genes against colistin, a last-resort antibiotic in human medicine for treating clinical infections caused by multidrug-resistant Gram-negative bacteria. Quantitative PCR assays we developed showed that 34.1% of (86/252) and 51.1% of (23/45) isolates were positive for the gene, 1.2% of (3/252) and 2.2% of (2.2%, 1/45) isolates were positive for , and 5.2% of (13/252) and 44.4% of (20/45) isolates were positive for Overall, 4.8% (9/189) of bacteria isolated from the flies were positive for the gene (: 8.3%, 4/48; : 12.5%, 1/8; : 11.8%, 2/17; : 4.9%, 2/41), while none were positive for and Four -positive isolates (two and two ) from blow flies trapped near a dumpster had a MIC for colistin above 4 mg/ml. This study reports carriage in spp. and detection of and after their initial identification in Belgium and China, respectively. This study suggests that flies might contribute significantly to the dissemination of bacteria, carrying these genes into a large variety of ecological niches. Further studies are warranted to explore the roles that flies might play in the spread of colistin resistance genes. Antimicrobial resistance is recognized as one of the most serious global threats to human health. An option for treatment of the Gram-negative ESKAPE (, , , , , and species) bacteria with multiple drug resistance was the reintroduction of the older antibiotic colistin. However, a mobile colistin resistance gene () has recently been found to occur widely; very recently, two other colistin resistance genes ( and ) have been identified in Belgium and China, respectively. In this study, we report the presence of colistin resistance genes in flies. This study also reports the carriage of colistin resistance genes in the genus and detection of and after their initial identification. This study will stimulate more in-depth studies to fully elucidate the transmission mechanisms of the colistin resistance genes and their interaction.
Topics: Animals; Anti-Bacterial Agents; Bacterial Proteins; China; Colistin; Diptera; Drug Resistance, Bacterial; Enterobacter cloacae; Enterobacteriaceae; Escherichia coli; Escherichia coli Proteins; Houseflies; Phylogeny; Providencia
PubMed: 29030447
DOI: 10.1128/AEM.01736-17 -
Frontiers in Microbiology 2020Symbiotic bacterial communities that colonize the digestive tract of tephritid fruit flies interact with nutrient intake to improve the flies' fitness and immunity. Some...
Symbiotic bacterial communities that colonize the digestive tract of tephritid fruit flies interact with nutrient intake to improve the flies' fitness and immunity. Some bacterial species consistently inhabit the tephritid guts and are transmitted to the next generation vertically. These species contribute significantly to some aspects of their host's physiology. In the current study, we examined the role of four vertically transmitted bacteria (, , , and ) on the fitness parameters and immunity of larvae that were fed a nutritionally manipulated diet. For this purpose, eggs were collected from axenic, gnotobiotic, and symbiotic adult flies, and larvae were reared on four types of diets in which carbohydrate and/or protein contents were reduced and then compared with larvae raised on a control diet. The diet and bacterial interactions significantly affected the fitness and immunity of . Larvae of axenic flies grew slower and displayed weaker immune-based responses (PO activity, antibacterial activity, survival) than larvae of gnotobiotic and symbiotic flies. Overall, larvae reared on the low-protein diet grew slower than those reared on the control or low-carbohydrate diets. Survival, PO activity, and antibacterial activity were significantly lower in the hemolymph of larvae reared on low-protein diets. Our results also revealed that the levels of hemolymph protein, glucose, trehalose, and triglyceride in larvae from axenic flies were significantly lower than those in larvae of the symbiotic group after they fed on most of the tested diets. These results strongly infer that diet and vertically transmitted bacteria are both essential contributors to the fitness and immunity of .
PubMed: 33193277
DOI: 10.3389/fmicb.2020.596352 -
Clinical Microbiology and Infection :... Sep 2018A carbapenem-resistant Providencia rettgeri (PR1) isolate was recovered from a wound infection in Missouri, USA. This isolate possessed an EDTA-inhibitable carbapenemase...
OBJECTIVES
A carbapenem-resistant Providencia rettgeri (PR1) isolate was recovered from a wound infection in Missouri, USA. This isolate possessed an EDTA-inhibitable carbapenemase that was unidentified using the Xpert CARBA-R assay. Our objective was to elucidate the molecular determinant of carbapenem resistance in this isolate. We then sought to test the transmissibility of bla loci in clinical P. rettgeri and Proteus mirabilis isolates.
METHODS
In October 2016 the novel ambler Class B carbapenemase bla, was reported in two different Proteus mirabilis (PM185 and PM187) isolates. Broth mating assays for transfer of carbapenemase activity were performed for the three clinical isolates with recipient sodium azide-resistant Escherichia coli J53. Antibiotic susceptibility testing and phenotypic carbapenemase activity testing were performed on the clinical isolates, J53 and transconjugants using the Kirby-Bauer disc diffusion method according to CLSI guidelines. Plasmid DNA from PM187, PR1 and their transconjugants were used as input for Nextera Illumina sequencing libraries and sequenced on a NextSeq platform.
RESULTS
PR1 was resistant to both imipenem and meropenem. PM187 and PR1 could transfer resistance to E. coli through plasmid conjugation (pPM187 and pPR1). pPM187 had a virB/virD4 type IV secretion system whereas pPR1 had a traB/traD type IV secretion system.
CONCLUSION
Two of three bla-bearing clinical isolates tested could conjugate resistance into E. coli. The resulting transconjugants became positive for phenotypic carbapenemase production but did not pass clinical resistance breakpoints. bla can be transmitted on different plasmid replicon types that rely on distinct classes of type IV secretion system for horizontal transfer.
Topics: Anti-Bacterial Agents; Bacterial Proteins; Disk Diffusion Antimicrobial Tests; Gene Transfer, Horizontal; High-Throughput Nucleotide Sequencing; Humans; Imipenem; Meropenem; Plasmids; Proteus mirabilis; Providencia; Sequence Analysis, DNA; Thienamycins; beta-Lactamases
PubMed: 29496594
DOI: 10.1016/j.cmi.2018.02.018 -
Microorganisms Jul 2023This study aimed to characterize the pathogenicity of bacteria isolated from the starter of two traditional beers produced and consumed in Benin. After standard...
This study aimed to characterize the pathogenicity of bacteria isolated from the starter of two traditional beers produced and consumed in Benin. After standard microbial identification, species were identified by specific biochemical tests such as catalase, coagulase, and API 20 E. Antibiotic sensitivity was tested according to the French Society of Microbiology Antibiogram Committee. The crystal violet microplate technique evaluated the biofilm production and conventional PCR was used to identify genes encoding virulence and macrolide resistance. According to our data, the traditional starter known as that is used to produce beer is contaminated by and staphylococci species. Thus, 28.43% of the isolated bacteria were coagulase-negative staphylococci (CNS), and 10.93% coagulase-positive staphylococci (CPS). Six species such as (1.38%), (4.14%) (5.51%) (6.89%) (15.16%), and (27.56%) were identified among . Those bacterial strains are multi-resistant to conventional antibiotics. The hight capability of produced biofilms was recorded with , (100%), (75%), and spp (60%). (4%) and coagulase-negative (5.55%) harbor the macrolide resistance gene. For other strains, these genes were not detected. Foods contaminated with bacteria resistant to antibiotics and carrying a virulence gene could constitute a potential public health problem. There is a need to increase awareness campaigns on hygiene rules in preparing and selling these traditional beers.
PubMed: 37630499
DOI: 10.3390/microorganisms11081939 -
Current Biology : CB Nov 2020Finding a suitable oviposition site is a challenging task for a gravid female fly, because the hatched maggots have limited mobility, making it difficult to find an...
Finding a suitable oviposition site is a challenging task for a gravid female fly, because the hatched maggots have limited mobility, making it difficult to find an alternative host. The oriental fruit fly, Bactrocera dorsalis, oviposits on many types of fruits. Maggots hatching in a fruit that is already occupied by conspecific worms will face food competition. Here, we showed that maggot-occupied fruits deter B. dorsalis oviposition and that this deterrence is based on the increased β-caryophyllene concentration in fruits. Using a combination of bacterial identification, volatile content quantification, and behavioral analyses, we demonstrated that the egg-surface bacteria of B. dorsalis, including Providencia sp. and Klebsiella sp., are responsible for this increase in the β-caryophyllene contents of host fruits. Our research shows a type of tritrophic interaction between micro-organisms, insects, and insect hosts, which will provide considerable insight into the evolution of insect behavioral responses to volatile compounds.
Topics: Animals; Behavior, Animal; Female; Fruit; Host Specificity; Host-Parasite Interactions; Klebsiella; Larva; Oviposition; Ovum; Polycyclic Sesquiterpenes; Providencia; Smell; Tephritidae; Volatile Organic Compounds
PubMed: 32946751
DOI: 10.1016/j.cub.2020.08.080 -
PloS One 2012Rhomboids are ubiquitous proteins with unknown roles in mycobacteria. However, bioinformatics suggested putative roles in DNA replication pathways and metabolite...
BACKGROUND
Rhomboids are ubiquitous proteins with unknown roles in mycobacteria. However, bioinformatics suggested putative roles in DNA replication pathways and metabolite transport. Here, mycobacterial rhomboid-encoding genes were characterized; first, using the Providencia stuartii null-rhomboid mutant and then deleted from Mycobacterium smegmatis for additional insight in mycobacteria.
METHODOLOGY/PRINCIPAL FINDINGS
Using in silico analysis we identified in M. tuberculosis genome the genes encoding two putative rhomboid proteins; Rv0110 (referred to as "rhomboid protease 1") and Rv1337 ("rhomboid protease 2"). Genes encoding orthologs of these proteins are widely represented in all mycobacterial species. When transformed into P. stuartii null-rhomboid mutant (ΔaarA), genes encoding mycobacterial orthologs of "rhomboid protease 2" fully restored AarA activity (AarA is the rhomboid protein of P. stuartii). However, most genes encoding mycobacterial "rhomboid protease 1" orthologs did not. Furthermore, upon gene deletion in M. smegmatis, the ΔMSMEG_4904 single mutant (which lost the gene encoding MSMEG_4904, orthologous to Rv1337, "rhomboid protease 2") formed the least biofilms and was also more susceptible to ciprofloxacin and novobiocin, antimicrobials that inhibit DNA gyrase. However, the ΔMSMEG_5036 single mutant (which lost the gene encoding MSMEG_5036, orthologous to Rv0110, "rhomboid protease 1") was not as susceptible. Surprisingly, the double rhomboid mutant ΔMSMEG_4904-ΔMSMEG_5036 (which lost genes encoding both homologs) was also not as susceptible suggesting compensatory effects following deletion of both rhomboid-encoding genes. Indeed, transforming the double mutant with a plasmid encoding MSMEG_5036 produced phenotypes of the ΔMSMEG_4904 single mutant (i.e. susceptibility to ciprofloxacin and novobiocin).
CONCLUSIONS/SIGNIFICANCE
Mycobacterial rhomboid-encoding genes exhibit differences in complementing aarA whereby it's only genes encoding "rhomboid protease 2" orthologs that fully restore AarA activity. Additionally, gene deletion data suggests inhibition of DNA gyrase by MSMEG_4904; however, the ameliorated effect in the double mutant suggests occurrence of compensatory mechanisms following deletion of genes encoding both rhomboids.
Topics: Anti-Bacterial Agents; Bacterial Proteins; Biofilms; Ciprofloxacin; Drug Resistance, Bacterial; Gene Knockout Techniques; Genetic Complementation Test; Mycobacterium smegmatis; Novobiocin; Peptide Hydrolases; Phenotype; Providencia
PubMed: 23029216
DOI: 10.1371/journal.pone.0045741 -
Mikrobiyoloji Bulteni Jul 2018Our country is the epicenter of the OXA-48-like carbapenemase-producing Klebsiella and Escherichia coli; and in the recent years, the concern has been increasing due to...
Our country is the epicenter of the OXA-48-like carbapenemase-producing Klebsiella and Escherichia coli; and in the recent years, the concern has been increasing due to both spreading of this resistance to other members of Enterobacteriaceae family and acquiring other carbapenemases by the OXA-48-producing strains. In this study, OXA-48 and NDM-1 co-production was presented in Providencia rettgeri. Two P.rettgeri strains that were resistant to all antimicrobials except colistin and tigecyclin, were isolated from two patients in the burn unit of our hospital, including one from the urine sample of a 68 years female in April 2017, and the other from a burn wound swab of a 35 years old male, in November 2017. Minimal inhibitory concentrations (MICs) of the isolates for imipenem and meropenem were measured as ≥ 32 µg/ml; and for colistin and tigecyclin were 1 ve 0.5 µg/ml, respectively. Multiplex PCR analysis showed that both strains were carrying and carbapenemases, and extended spectrum beta-lactamase genes. By using DNA sequence analysis, the TEM gene was typed as blaTEM-1. The Pulsed Field Gel Electrophoresis (PFGE) analysis indicated that these two strains which were consecutively isolated from two different patients in a single unit within about seven months were genetically indistinguishable. No significant data that could explain the spread of these isolates was obtained from our retrospective analysis of the medical records including the results of environmental surveillance cultures, and patients' history. Nevertheless, hospital infection control committee enforced the infection control measures in that unit, and no further isolation was observed within three months period following the last isolation, neither from environmental nor from clinical samples. With this study, it was emphasized that the co-production of OXA-48 and NDM-1 carbapenemases which was reported from only three Enterobacteriaceae species up to date was ongoing for spreading to other species by using horizontal route, and also showing a potential to be a growing problem in the hospitals, by clonal expansion (vertical route). Effectively using of the molecular epidemiological methods will provide useful data to better understand the transmission dynamics of such rare, but problematic species in hospitals.
Topics: Adult; Anti-Bacterial Agents; Bacterial Proteins; Enterobacteriaceae Infections; Female; Humans; Male; Microbial Sensitivity Tests; Providencia; Retrospective Studies; beta-Lactamases
PubMed: 30156516
DOI: 10.5578/mb.67153 -
Antimicrobial Agents and Chemotherapy Jan 2012qnr genes are plasmid-mediated quinolone resistance genes mainly harbored on large conjugative multiresistant plasmids. The qnrD gene was recently observed in Salmonella...
qnr genes are plasmid-mediated quinolone resistance genes mainly harbored on large conjugative multiresistant plasmids. The qnrD gene was recently observed in Salmonella enterica on a small nonconjugative plasmid (p2007057). We describe two strains of Providencia rettgeri harboring qnrD on nonconjugative plasmids. The plasmids were 99% identical, with 2,683 bp and four open reading frames, including qnrD, but exhibited only 53% identity with the plasmid found in S. enterica.
Topics: Anti-Bacterial Agents; Base Sequence; DNA, Bacterial; Drug Resistance, Bacterial; Enterobacteriaceae Infections; Feces; Humans; Microbial Sensitivity Tests; Molecular Sequence Data; Open Reading Frames; Plasmids; Providencia; Quinolones; Salmonella Infections; Salmonella enterica; Sequence Homology, Nucleic Acid
PubMed: 21986831
DOI: 10.1128/AAC.00081-11