scholarly journals Distinction between Antimicrobial Resistance and Putative Virulence Genes Characterization in Plesiomonas shigelloides Isolated from Different Sources

Antibiotics ◽  
2022 ◽  
Vol 11 (1) ◽  
pp. 85
Author(s):  
Samy Selim ◽  
Mohammed S. Almuhayawi ◽  
Shadi Ahmed Zakai ◽  
Ahmed Attia Salama ◽  
Mona Warrad

Plesiomonas shigelloides are gram-negative, thermotolerant, motile, and pleomorphic microorganisms that are only distantly related to those of the Enterobacteriaceae and Vibrionaceae families. One of the most common sources of P. shigelloides contamination is human stool, but it may also be found in a wide range of other animals, plants, and aquatic habitats. Antimicrobial resistance in P. shigelloides from seawater and shellfish was investigated, and pathogenicity involved genes were characterized as part of this study. Out of 384 samples of shellfish, 5.7% included P. shigelloides. The presence of P. shigelloides was also discovered in 5% of the seawater sampled. The antimicrobial resistance of 23 P. shigelloides isolates derived from those samples was investigated. All isolates were sensitive to nalidixic acid, carbenicillin, cephalothin, erythromycin, kanamycin, tetracycline, and ciprofloxacin in the study. Several strains isolated from diseased shellfish were tested for virulence in shellfish by intraperitoneal injections. The LD50 values ranged from 12 × 108 to 3 × 1012 cfu/shellfish. When looking for possible virulence factors that may play a significant role in bacterial infection in the current study, we found that all of these genes were present in these strains. These include genes such as elastase, lipase, flagellin, enterotoxin, and DNases. According to these findings, shellfish may serve as a reservoir for multi-resistant P. shigelloides and help spread virulence genes across the environment.

2018 ◽  
Author(s):  
Mahmoud M. Ashawesh ◽  
Robert Markus ◽  
Christopher N. Penfold ◽  
Kim R. Hardie

AbstractBacterial infection of humans, animals and plants relies heavily on secreted proteases that degrade host defences or activate bacterial toxins. The largest family of proteins secreted by Gram-negative pathogenic bacteria, the Autotransporters (ATs), includes key proteolytic virulence factors. There remains uncertainty about the mechanistic steps of the pathway ATs share to exit bacteria, and how it is energetically driven. This study set out to shed light on the AT secretion pathway with the ultimate aim of uncovering novel antimicrobial targets that would be unlikely to trigger the development of resistance mechanisms in bacteria. To do this, two AT virulence factors with distinct proteolytic functions, EspC (secreted from EnteropathogenicEscherichia coli) and AaaA (tethered to the extracellular surface ofPseudomonas aeruginosa) were chosen. EspC and AaaA were fluorescently labelled using two separate methods to establish the localization patterns of ATs as they are secreted from a bacterial cell. Super resolution microscopy revealed that localization of ATs occurs via a helical route along the bacterial cytoskeleton. In addition to requiring the conserved C-terminal β-barrel translocator domain of the AT, we present the first evidence that secretion is dependent on a dynamic interaction with a structure reliant upon the actin homologue MreB and the Sec translocon. These findings provide a step forward in the mechanistic understanding of the secretion of this widely distributed family of proteins that have pivotal roles in bacterial pathogenesis and conserved structural properties that could serve as novel broad-range antimicrobial targets.SignificanceSecreted bacterial proteases facilitate the infection of human, animal and plant hosts by degrading host defences or activating bacterial toxins. The autotransporter family is the largest family of proteins secreted from Gram-negative bacteria, and includes proteolytic virulence factors crucial to bacterial infection. Precisely how autotransporters migrate from the inside to the outside of the cell, and how this movement is energetically driven is a mystery. We demonstrate a spiral pathway of autotransporter secretion, presenting evidence that it involves a dynamic interaction with the actin homologue MreB that comprises the bacterial cytoskeleton. Our findings open the way to unravelling the mechanism of autotransporter secretion and offer the possibility to identify novel antimicrobial targets unlikely to trigger the development of antimicrobial resistance.


2019 ◽  
Author(s):  
Gustavo Enck Sambrano ◽  
Gustavo P Riboldi ◽  
Keli C Reiter ◽  
Thiago Galvão da Silva Paim ◽  
Neidmar Correa Tolfo ◽  
...  

Background: Streptococcus pyogenes, a Group A streptococci (GAS), is an important human pathogen that causes a wide range of infections. Methods: Twenty five clinical isolates of S. pyogenes were submitted to an emm typing and to a Real-time PCR analysis for 23 important virulence factors. Results: Fourteen emm types were found and the emm1 type was the most prevalent. The majority of the isolates were classified as emm pattern E, followed by A-C3. No pattern D was found. Among the virulence factors, the most prevalent were SpeG, Slo, C5a-peptidase and SPNA. Phage encoded virulence genes were also found among the strains, such as mf-2, SpeJ and SpeL. Discussion: The emm1 type was the most prevalent while the 13 others M types were distributed along the strains. No tissue tropism was found on the isolates. The virulence factors analysis demonstrated that chromosomally and phage-encoded genes were found, which confers a potential for high virulent micro-organisms.


2019 ◽  
Author(s):  
Gustavo Enck Sambrano ◽  
Gustavo P Riboldi ◽  
Keli C Reiter ◽  
Thiago Galvão da Silva Paim ◽  
Neidmar Correa Tolfo ◽  
...  

Background: Streptococcus pyogenes, a Group A streptococci (GAS), is an important human pathogen that causes a wide range of infections. Methods: Twenty five clinical isolates of S. pyogenes were submitted to an emm typing and to a Real-time PCR analysis for 23 important virulence factors. Results: Fourteen emm types were found and the emm1 type was the most prevalent. The majority of the isolates were classified as emm pattern E, followed by A-C3. No pattern D was found. Among the virulence factors, the most prevalent were SpeG, Slo, C5a-peptidase and SPNA. Phage encoded virulence genes were also found among the strains, such as mf-2, SpeJ and SpeL. Discussion: The emm1 type was the most prevalent while the 13 others M types were distributed along the strains. No tissue tropism was found on the isolates. The virulence factors analysis demonstrated that chromosomally and phage-encoded genes were found, which confers a potential for high virulent micro-organisms.


2017 ◽  
Vol 07 (04) ◽  
pp. 016-020
Author(s):  
Juliet Roshini Mohan Raj ◽  
Rajeshwari Vittal ◽  
Santosh Kogaluru Shivakumaraswamy ◽  
Vijaya Kumar Deekshit ◽  
Indrani Karunasagar

AbstractDownstream water systems provide for a conducive environment for horizontal gene transfer. The objective of this study was to determine the burden of antimicrobial resistance in waste water effluents from different sources and their impact on human health. Gram negative bacteria were isolated from 30 samples each of industrial, hospital and domestic effluents. The antimicrobial susceptibility of the 367 isolates from 90 effluent samples was determined by disc diffusion test and presence of antimicrobial resistance genes by polymerase chain reaction. Resistance to ampicillin was 62% in hospital effluents and was higher than that recorded for industrial and domestic effluents. While the highest percentage of resistance to tetracycline was observed in isolates from industrial effluents (42%) a low of 9.5% was observed in hospital effluents. Antimicrobial resistance determinants present on mobile genetic elements were observed in a small fraction (~10%) of the resistant isolates. The resistance profile of isolates in effluents reflect the practices of different industries. Resistant isolates in domestic effluents could be a reflection of the indiscriminate use of antibiotics andthat many of the contents of disinfectants and cleaning agents routinely used may contain structural analogs of antimicrobials used in therapy. Though by phenotypic test a higher prevalence of antimicrobial resistance was recorded the genotypic study revealed the prevalence to be low. This could be due to the limited number of antimicrobial resistance genes included in this study.


Author(s):  
N. Sujatha ◽  
K. Lakshmi Kavitha ◽  
K.V. Subramanyam ◽  
T. Srinivasa Rao ◽  
R.N. Ramani Pushpa

Background: Pasteurella multocida is the causative agent of many economically important diseases in a wide range of hosts. The mechanisms by which these bacteria can invade the mucosa, evade innate immunity and cause systemic disease are slowly being elucidated. Many key virulence factors are yet to be identified, including those required for initial attachment and invasion of host cells and for persistence in a relatively nutrient poor and hostile environment. This has led to intensive research to understand host adaptation mechanisms and virulence factors in order to develop effective vaccines. Methods: The present study was carried out to know the distribution of virulence genes viz., haemoglobin binding proteins (hgbA and hgbB), outer membrane protein (ompH), fimbrial antigen (ptfA), filamentous haemagglutinin (pfhA) and transferrin binding protein (tbpA) by PCR in P. multocida CapA isolates from apparently healthy or carrier animals and CapB isolates from field Haemorrhagic septicemia (HS) cases to monitor the epidemiological associations of virulence genes in Cap A and Cap B isolates.Result: The study revealed that all the six virulence associated genes were present in Cap B isolates. None of the Cap A isolates harboured tbpA and pfhA genes. These two genes were closely related to serotype B causing Haemorrhagic septicemia and were epidemiologically associated with disease status.


2021 ◽  
Vol 12 (3) ◽  
pp. 698-710
Author(s):  
Peter Rabinowitz ◽  
Bar Zilberman ◽  
Yair Motro ◽  
Marilyn C. Roberts ◽  
Alex Greninger ◽  
...  

Brucellosis has a wide range of clinical severity in humans that remains poorly understood. Whole genome sequencing (WGS) analysis may be able to detect variation in virulence genes. We used Brucella melitensis sequences in the NCBI Sequence Read Archive (SRA) database to assemble 248 whole genomes, and additionally, assembled 27 B. melitensis genomes from samples of human patients in Southern Israel. We searched the 275 assembled genomes for the 43 B. melitensis virulence genes in the Virulence Factors of Pathogenic Bacteria Database (VFDB) and 10 other published putative virulence genes. We explored pan-genome variation across the genomes and in a pilot analysis, explored single nucleotide polymorphism (SNP) variation among the ten putative virulence genes. More than 99% of the genomes had sequences for all Brucella melitensis virulence genes included in the VFDB. The 10 other virulence genes of interest were present across all the genomes, but three of these genes had SNP variation associated with particular Brucella melitensis genotypes. SNP variation was also seen within the Israeli genomes obtained from a small geographic region. While the Brucella genome is highly conserved, this novel and large whole genome study of Brucella demonstrates the ability of whole genome and pan-genome analysis to screen multiple genomes and identify SNP variation in both known and novel virulence genes that could be associated with differential disease virulence. Further development of whole genome techniques and linkage with clinical metadata on disease outcomes could shed light on whether such variation in the Brucella genome plays a role in pathogenesis.


Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1724
Author(s):  
Hyo Jung Kang ◽  
Sunghyun Yoon ◽  
Koeun Kim ◽  
Young Ju Lee

Enterococci, which are considered environmental mastitis-causing pathogens, have easily acquired aminoglycoside-resistant genes that encode various aminoglycoside-modifying enzymes (AME). Therefore, this study was conducted to compare the distribution of high-level aminoglycoside-resistant (HLAR) and multidrug-resistant (MDR) Enterococcus faecalis (E. faecalis) bacteria isolated from bulk tank milk in four dairy companies in Korea. Moreover, it analyzed the characteristics of their antimicrobial resistance genes and virulence factors. Among the 301 E. faecalis bacteria studied, 185 (61.5%) showed HLAR with no significant differences among the dairy companies. Furthermore, 129 (69.7%) of the 185 HLAR E. faecalis showed MDR without significant differences among companies. In contrast, HLAR E. faecalis from companies A, B, and C were significantly higher in resistance to the four classes than those in company D, which had the highest MDR ability against the three antimicrobial classes (p < 0.05). In addition, in the distribution of AME genes, 72 (38.9%) and 36 (19.5%) of the isolates carried both aac(6′)Ie-aph(2″)-la and ant(6)-Ia genes, and the ant (6)-Ia gene alone, respectively, with significant differences among the companies (p < 0.05). In the distribution of virulence genes, the ace (99.5%), efa A (98.9%), and cad 1 (98.4%) genes were significantly prevalent (p < 0.05). Thus, our results support that an advanced management program by companies is required to minimize the dissemination of antimicrobial resistance and virulence factors.


2018 ◽  
Vol 46 (1) ◽  
pp. 6
Author(s):  
Carolina Maciel Malgarin ◽  
Karine Ludwig Takeuti ◽  
Anne Caroline de Lara ◽  
David Emilio Santos Neves de Barcellos

Background: One of the most frequent health problems in the swine industry is the post-weaning diarrhea in nursery pigs, which leads to significant losses due to weight loss, dehydration, cost of medication and mortality. Escherichia coli  (E. coli) is one of the main bacterial agents of the post-weaning diarrhea. To investigate the possibility of enterotoxigenic  E. coli (ETEC) transmission through drinking water to nursery piglets, the objective of this study was to isolate, characterize by virulence factors, and compare the antimicrobial resistance profiles of E. coli from drinking water samples in nurseries and from rectal swabs of their piglets presenting post-weaning colibacillosis.Materials, Methods & Results: Fifteen rectal swabs from diarrheic piglets in their first three weeks after weaning and one water sample were collected from each of ten nurseries located in Rio Grande do Sul State, south of Brazil. After enrichment with a commercial broth medium, water samples were cultured in blood agar, as well as the rectal swab samples, and the characteristic colonies were identified by standard biochemical analysis. Following isolation and identification of E. coli, the colonies from water samples and their corresponding piglets’ samples were characterized by multiplex PCR in order to determine specific ETEC fimbria and toxin genes. Finally, all E. coli isolates were submitted to antimicrobial susceptibility testing. Virulence factors and antimicrobial sensitivity could then be compared between water and piglets’ samples. The difference in the antimicrobial resistance frequency for each of the sample groups were compared using the multi comparison test. E. coli was isolated in four out of the ten water samples, although none of the water samples presented ETEC virulence factors. From 60 rectal swab samples (15 from each of the four positive farms with E. coli isolated from water samples), 21 E. coli were isolated and seven demonstrated characteristic ETEC virulence factors. The fimbriae exhibited in higher frequency were F18 (62.5%) and F4 (25%) and the toxins were STb (100%) and STaP (75%). E. coli isolated from water samples presented higher resistance to the antimicrobials apramycin, florfenicol, lincomycin, lincomycin+spectinomycin, oxytetracycline, and sulfamethoxazole+trimethoprim; it did not present resistance to colistin and fosfomycin. The seven ETEC from rectal swab samples presented a higher resistance to lincomycin, and lower resistance frequency to fosfomycin. The other 14 E. coli non-ETEC from rectal swab samples presented a higher resistance to florfenicol and no resistance to colistin.Discussion: Enterotoxigenic E. coli is an important agent causing post-weaning colibacillosis, although, differently from other studies, this experiment did not find the agent in most of the sampled animals. In contrast to other authors, ETEC was not found in water, as the development of its virulence factors may depend on conditions presented exclusively in the animal. By the results we can conclude that, although E. coli was isolated from the drinking water, it was not a significant mechanism for nursery piglets’ infection with ETEC in this experiment. The samples analyzed presented a wide range of resistance to different antimicrobials, including multi-resistance. In some cases, E. coli found in water presented different antimicrobial profile from the bacterium found in the rectal swab samples. Enterotoxigenic E. coli was susceptible to fosfomycin and its use may represent a prudent antimicrobial choice to the swine industry.


Author(s):  
Mohamed Sabry Abd Elraheam Elsyaed ◽  
Mary Mounir

Aims: A great concern directed to non-O157 Shiga toxin-producing Escherichia coli (STEC) serotypes due to their public health importance. Detecting the existence, antimicrobial profiles, and virulence repertoire of different STEC serotypes from animals essential for human food are important. Study Design: This study aimed to investigate the presence of STEC in different hosts, the distribution pattern of stx1, stx2, eaeA, and hlyA genes encoding Shiga toxins 1 and 2, intimin, and enterohemolysin, respectively, and the antimicrobial resistance of the detected serotypes. Results: A total of 75 samples were collected, 20 fecal samples from broilers, 15 fecal samples from ducks, 20 beef samples, and 20 human urine samples. Escherichia coli was detected at a rate of 60/75 (80%) distributed as;17 (85%), 8 (53.3%), 15(75%), and 20 (100%) from broilers, ducks, human urine, and beef samples, respectively. There was a significant difference between the isolation rates of E. coli from different sources with p<0.05. The prevalent serotypes were O78, O2:H6, O15:H2, and O26: H1. The frequency of stx1 gene was 56/60 (93.3%), stx2 gene was 55/60 (91.6%), the eaeA gene was 35/60 (58.3%), and hlyA gene was 26/60 (43.3%). The most effective antimicrobials were amoxicillin/clavulanic acid and ampicillin with the efficacy of 52/60 (86.6%) for each, while the efficacy of chloramphenicol and doxycycline was 5/60 (8.3%) and 44/60 (73.3%), respectively. Conclusion: The high frequency of non-O157 Shiga toxin-Producing Escherichia coli from different samples at Sadat City, high prevalence of virulence factors and resistance to chloramphenicol and doxycycline will help in monitoring the distribution of virulent serotypes and contribute to the establishment of control measures to reduce the spread of infection.


2015 ◽  
Vol 60 (3) ◽  
pp. 1874-1877 ◽  
Author(s):  
S. Baron ◽  
S. Delannoy ◽  
S. Bougeard ◽  
E. Larvor ◽  
E. Jouy ◽  
...  

This study investigated antimicrobial resistance, screened for the presence of virulence genes involved in intestinal infections, and determined phylogenetic groups ofEscherichia coliisolates from untreated poultry and poultry treated with ceftiofur, an expanded-spectrum cephalosporin. Results show that none of the 76 isolates appeared to be Shiga toxin-producingE. colior enteropathogenicE. coli. All isolates were negative for the major virulence factors/toxins tested (ehxA,cdt, heat-stable enterotoxin [ST], and heat-labile enterotoxin [LT]). The few virulence genes harbored in isolates generally did not correlate with isolate antimicrobial resistance or treatment status. However, some of the virulence genes were significantly associated with certain phylogenetic groups.


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