Plesiomonas Shigelloides Isolates Research Articles

Pathogenic characteristics of an aggregated diarrhea event caused by Plesiomonas shigelloides from stream.

This study aimed to investigate the cause of a foodborne disease outbreak in Huzhou on August 14, 2023. Multiple enteropathogens were detected using FilmArray, and the pathogen was subsequently isolated and cultured from anal swabs of the cases and stream water. The isolated strains were identified using VITEK MS, and antimicrobial susceptibility test, pulsed field gel electrophoresis (PFGE) molecular typing, and whole genome sequencing (WGS) were performed on the isolates of Plesiomonas shigelloides. Gene annotation and sequence alignment were used to analyze the virulence genes and drug resistance genes of the strains. A phylogenetic tree was constructed based on single nucleotide polymorphism (SNP), and homology analysis was conducted to trace the origin of P. shigelloides. A total of 7 strains of P.shigelloides were isolated, with 3 from stream water and 4 from anal swabs. All 7 strains exhibited the same PFGE pattern and showed resistance to amikacin, trimethoprim-sulfamethoxazole, chloramphenicol, tetracycline, cefazolin, streptomycin, and florfenicol. The isolated strains carried the same resistance genes and virulence factors. In the sequences of the isolated strains from this outbreak, 11 mutation sites were detected. The phylogenetic tree based on SNP sites showed that these strains were homologous. This foodborne disease outbreak caused by P.shigelloides was the first reported in Huzhou. WGS can be used as a complementary method to PFGE for epidemiological investigations of disease outbreaks.

Antimicrobial resistance in freshwater Plesiomonas shigelloides isolates: Implications for environmental pollution and risk assessment

Antibiotic resistance is known to impact treatment efficiency of Plesiomonas infections negatively with fatal outcomes. This study investigated antibiogram fingerprint of P. shigelloides (n = 182) isolated from three South Africa rivers using the disc diffusion technique. Environmental pollution and analogous health risk (given infections) that could associate with the freshwaters and empirical treatment of Plesiomonas were assessed using Antibiotic Resistance Index (ARI) and Multiple Antibiotic Resistance Indices (MARI), respectively. Thirteen EUCAST recommended (ERAs) and eleven non-recommended antibiotics (NAs) used as first line agents in the treatment of gastroenteritis and extraintestinal infections were tested. Resistance against ERAs decreased from cefoxitin (37.91%), cefuroxime (35.17%), cefepime (31.87%), ceftriaxone (29.67%), ciprofloxacin (18.13%), trimethoprim-sulfamethoxazole (10.44%), piperacillin/tazobactam (8.79%), ertapenem (4.95%), norfloxacin (4.40%), levofloxacin (2.75%), meropenem (1.10%) to imipenem (0.55%). The isolates had higher resistance (≥36.07%) against NAs but were susceptible to amikacin (67.58%), gentamycin (73.08%), and tetracycline (80.77%). MARI of the isolates were significantly different between ERAs and NAs (P-value < 0.05) and had an average of 0.17 ± 0.18 and 0.45 ± 0.13, respectively. About 33.87% and 95.63% of the isolates had MARI value from 0.23 to 0.62 and 0.27–0.82 to ERAs and NAs, respectively. Also, ERAs-based and NAs-based ARI across sampling units showed significantly different (P-value < 0.05) means of 0.18 ± 0.09 and 0.46 ± 0.05, respectively. MARI attributed low risk of empirical treatment to recommended antibiotics but higher risk to non-recommended antibiotics. Model estimated successful and unsuccessful empirical treatment of infections risks due to resistance in the isolates using recommended antibiotics as 65.93% and 34.07%, respectively; 1.65% and 98.35% in the case of non-recommended antibiotics, respectively. ARI based on recommended antibiotics identified potential environmental pollutions in a number of sites. Resistance in freshwater P. shigelloides especially against cephalosporin, quinolones and fluoroquinolones is distressing and might suggests high pollution of the freshwaters in the Eastern Cape Province.

Vibriosis and Aeromonas infection in shrimp: Isolation, sequencing, and control

Background and Aim: Shrimp is one of the most commonly consumed types of seafood. It is a very nutritious healthy food. Shrimp is low in calories and rich in protein and healthy fats. It also contains a treasure trove of vitamins and minerals. On the negative side, it may be affected by many bacterial diseases which affect its health. Furthermore, it may be incriminated as a vector of foodborne illnesses that range from mild gastrointestinal upset to life-threatening diseases. This study was designed to assess the clinical picture and zoonotic importance of vibriosis and Aeromonas infection in live shrimp and to study the antibacterial effect of citric acid (lemon juice) and acetic acid (vinegar) on these pathogens. Materials and Methods: A total of 170 live shrimp (Metapenaeus monoceros) samples were collected from Suez City, Egypt. The samples were examined clinically, and then, they were enriched into alkaline peptone water and cultivated on thiosulfate-citrate-bile salts-sucrose agar and ampicillin MacConkey agar for the isolation of Vibrio and Aeromonas species, respectively. The recovered isolates were confirmed biochemically and genotypically using duplex polymerase chain reaction (PCR) and sequencing. The germicidal effects of vinegar and lemon on artificially contaminated shrimp samples with Aeromonas hydrophila and Vibrio parahaemolyticus at different times (0.25, 1, 1.5, and 24 h) and temperatures (5° and 30°C) were studied. Results: The results revealed that some of the infected shrimp were hypoxic, lethargic with abnormal swimming behavior. In most cases, body appendages, telsons, uropods, and gills took black coloration. In addition, the hepatopancreas appeared soft, swollen, and congested. The prevalence rates of vibriosis in each of the musculature and hepatopancreas were 4.7%, while the prevalence rates of Aeromonas infection in the musculature and hepatopancreas were 11.8% and 11.2%, respectively. Duplex PCR showed that Aeromonas isolates gave double bands: 237 bp specific for gcat and 500 bp specific for 16S rRNA, while Vibrio spp. and Plesiomonas shigelloides isolates gave single band at 500 bp. The effect of organic acid treatment showed that acetic acid (vinegar 5%) had increasing reduction rates that reached its maximum level after 24 h; where it caused (100% inhibition) for A. hydrophila at both temperatures and (33.63% and 60% inhibition) for V. parahaemolyticus at refrigerator and room temperatures, respectively. Moreover, acetic acid was more effective at room temperature than at refrigerator temperature. Concerning the effect of lemon juice (citric acid), it was more effective than acetic acid at short marination (0.25 and 1 h) at both temperatures for the two pathogens. Moreover, lemon was more effective at refrigerator temperature than at room temperature at the same aforementioned time. The difference between the reduction effects of the two acids on both pathogens was statistically significant (p&lt;0.0001). Conclusion: Overall, the examined shrimp samples were found to be vectors for Vibrio and Aeromonas spp. Application of hygienic measures during handling and cooking of shrimp should be esteemed. The organic acid treatment trial showed that vinegar and lemon juice can be used as a safe and economic method to limit the microbial contamination in seafood.

Incidence and antimicrobial susceptibility fingerprints of Plesiomonas shigelliodes isolates in water samples collected from some freshwater resources in Southwest Nigeria

Plesiomonas shigelloides, is an emerging and significant enteric pathogen in water having implication in both localised and gastrointestinal infections with characteristic of displaying high resistance against commonly used antibiotics. This study evaluated the prevalence of Plesiomonas shigelloides and their antibiogram fingerprints in water sample collected from four rivers in South-western Nigeria. In all, 148 presumptive Plesiomonas shigelloides isolates was recovered from the rivers out of which 66 (44.6%) were confirmed positive for the organism using polymerase chain reaction techniques. Confirmed isolates were evaluated for their antibiogram profiles against a panel of 20 antimicrobials using the disc diffusion method and further screened for relevant antibiotic resistance genes. Resistance of the isolates against the antimicrobials followed the order: sulphamethoxazole (100%), erythromycin (93%), ampicillin (90%), cephalotin (82%), streptomycin (64%), and chloramphenicol (58%), amoxicillin (53%), cefotaxime (50%), tetracycline (49%), neomycin (38%) and trimethoprim + sulphamethoxazole (38%). Conversely, all the isolates were susceptible against netilmicin, and susceptibility against the other antibiotics follows the order: meropenem (94%), gentamicin (88%), imipenem (79%), amikacin (70%), ciprofloxacin (70%), norfloxacin (59%), trimethoprim (56%) and ceftazidine (56%). The multiple antibiotic resistance indices of the organism were higher than the accepted threshold of 0.2. The incidence of 11 antimicrobial resistance determinants were obtained as follows: [sulphonamides; (sulI (18%), sulII (20%), dfr1 (70%), dfr(18) (5%)), [beta-lactams; (ampC 37%)], [tetracyclines; (tetA (78%), tetE (57%)], [phenicols; (catII (16%), cmlA1 (11%)] and [aminoglycosides; (aphA2 (36%) and strA (67%)]. Pearson chi-square exact test revealed positive associations among tetA, tetE, sullI and catII and tetA genes. To the best of our knowledge, this is the first report on the incidence and antibiogram fingerprint of P. shigelloides in these freshwater resources and we conclude that these rivers are important reservoirs of multiple antimicrobial resistant biotypes of this organism, and consequently a threat to public health.

Assessment of Aquatic Aeromonas spp. Isolates' Susceptibility to Cinnamaldehyde, Vanillin, and Crude Kigelia africana Fruit Extracts

AbstractAeromonas spp. are primary and opportunistic fish pathogens that are responsible for severe economic losses in aquaculture, and are exacerbated by their increasing incidence of antimicrobial resistance. Phytochemicals are being explored as alternatives to conventional antimicrobial agents since they have destressing, growth‐promoting, immune‐stimulating, and antimicrobial properties. The susceptibility of 93 aquatic Aeromonas spp. and six Plesiomonas shigelloides isolates (from cultured catfish, koi carp, tilapia, and seawater) to three phytochemicals, viz.: cinnamaldehyde (10–1250 µg/mL), vanillin (5–250 µg/mL), and crude Kigelia africana fruit extracts (4–10 mg/mL ethyl acetate, dichloromethane, methanol, and hexane) were assessed using the disk diffusion assay and compared to ampicillin and tetracycline, using activity indices. No zones of inhibition were obtained with 10 µg/mL of cinnamaldehyde, or with 5–250 µg/mL of vanillin. However, varying degrees of inhibition were observed with higher concentrations of cinnamaldehyde, as well as with the K. africana methanol extract. Cinnamaldehyde (≥500 µg/mL) and the K. africana methanol extract displayed better antimicrobial activity against study isolates in comparison to vanillin and ampicillin. They appear to be promising and sustainable phytochemicals that might be useful as alternatives to the antimicrobial agents currently in use in aquaculture.

Occurrence of Plesiomonas shigelloides in Cultured Red Hybrid Tilapia (Oreochromis niloticus) from Tropical Rivers, East Coast Malaysia

A total of 182 isolates of Plesiomonas shigelloides were identified from 40 healthy red hybrid tilapia, Oreochromis niloticus cultured at two important rivers in Terengganu, Malaysia namely Como River and Terengganu River from east coast Malaysia. P. shigelloides count in Digestive Tract Content (DTC) and Muscle (MUS) of red hybrid tilapia cultured at Terengganu River was 1000-fold higher than Como River. Antibiotic susceptibility test was also performed on Plesiomonas shigelloides isolates. The incidence of antibiotic resistance was higher in Plesiomonas shigelloides isolated from red hybrid tilapia cultured at Terengganu River compared to Como river. Thus, the findings of the study indicate that P. shigelloides from tilapia muscle and an intestine could be an alarming for serious public health risk to consumers.

Phenotypic characterization and putative virulence factors of human, animal and environmental isolates of Plesiomonas shigelloides

Plesiomonas shigelloides (a bacterium widely distributed in aquatic ecosystems causing both intestinal and extra-intestinal diseases) shows a host of putative virulence markers, such as hemolysins, cytotoxins, production of exoenzymes associated with pathogenicity, adhesive ability and vacuolation of cell lines in vitro. Technical difficulties in detecting some of these virulence factors together with scantiness of epidemiological information, due to the lack of routine analysis for P. shigelloides as etiological agent of gastroenteritis, lead to sporadic and occasional finding of these bacteria. All this casts doubt on the real virulence potential of P. shigelloides and fuels a debate about assignment of these bacteria to the list of human pathogens. Here we demonstrated the phenotypic diversity and the putative virulence markers by examining serotype biochemical and virulence properties of 60 strains of P. shigelloides isolated from human, animal and environmental samples in different countries, which showed the unpredictable occurrence of the above properties depending on various locations and diverse sources.

Identification of Plesiomonas spp.: Serological and MALDI-TOF MS methods

Biochemical and serological profiles of isolates of Plesiomonas shigelloides were assayed using standard procedures in isolates from various clinical samples. Seventy-four isolates, including P. shigelloides type strain, were further characterized by MALDI-TOF MS using 3-methoxy-4-hydroxycinnamic acid as matrix. Multiple ions in the 3- to 12-kDa mass range were found in the spectra of each strain, from which the "species-identifying" unique biomarker ions were identified. After creating the species-specific patterns, a spectral database was generated for reliable, rapid, reproducible and accurate identification of Plesiomonas strains. The classical strain description (biochemical and serological) was thus complemented with the metabolic (proteomic) characterization.

Identification of human enteric pathogens in gull feces at Southwestern Lake Michigan bathing beaches

Ring-billed (Larus delawarensis Ord, 1815) and herring (Larus argentatus Pontoppidan, 1763) gulls are predominant species of shorebirds in coastal areas. Gulls contribute to the fecal indicator burden in beach sands, which, once transported to bathing waters, may result in water quality failures. The importance of these contamination sources must not be overlooked when considering the impact of poor bathing water quality on human health. This study examined the occurrence of human enteric pathogens in gull populations at Racine, Wisconsin. For 12 weeks in 2004 and 2005, and 7 weeks in 2006, 724 gull fecal samples were examined for pathogen occurrence on traditional selective media (BBL CHROMagar-Salmonella, Remel Campy-BAP, 7% horse blood agar) or through the use of novel isolation techniques (Campylobacter, EC FP5-funded CAMPYCHECK Project), and confirmed using polymerase chain reaction (PCR) for pathogens commonly harbored in gulls. An additional 226 gull fecal samples, collected in the same 12-week period in 2004, from a beach in Milwaukee, Wisconsin, were evaluated with standard microbiological methods and PCR. Five isolates of Salmonella (0.7%), 162 (22.7%) isolates of Campylobacter, 3 isolates of Aeromonas hydrophila group 2 (0.4%), and 28 isolates of Plesiomonas shigelloides (3.9%) were noted from the Racine beach. No occurrences of Salmonella and 3 isolates of Campylobacter (0.4%) were found at the Milwaukee beach. A subset of the 2004 samples was also examined for Giardia and Cryptosporidium and was found to be negative. Information as to the occurrence of human pathogens in beach ecosystems is essential to design further studies assessing human health risk and to determine the parameters influencing the fate and transport of pathogens in the nearshore environment.

Genetic Variability Among Isolates of Plesiomonas shigelloides from Fish, Human Clinical Sources and Fresh Water, Determined by RAPD Typing

Plesiomonas shigelloides has been a suspected pathogen for the past 40 years. Random amplified polymorphic DNA (RAPD) analysis with two random primers was used to examine the genetic diversity among 26 strains of P. shigelloides: 10 from fresh water, 6 from fish, and 10 from human clinical sources. There was notable genetic variability among most of the isolates, and none of the isolates had the same composite RAPD profile. Our results indicate that most of the isolates from the same source grouped together and that the isolates from fish had a closer linkage to the human clinical isolates than did the freshwater isolates, suggesting that fish may be the more serious source for potential risk of infection. We also found that certain human clinical isolates had almost the same RAPD profiles as certain of the isolates from freshwater and fish, indicating that P. shigelloides from both sea food and freshwater should be considered potential pathogens.

Serotypes and anti-microbial susceptibility of Plesiomonas shigelloides isolates from humans, animals and aquatic environments in different countries

A total of 73 strains of Plesiomonas shigelloides isolated from humans (24 strains) animals (21 strains) and aquatic environment (28 strains) were determined for their O:H serotype and susceptibility to 18 anti-microbial substances and to the vibriostatic agent O/129. Of all strains, 86.3% were typeable by the O and 94.5% by the H anti-sera used. The serotype distribution was heterogeneous within a country and between the countries. Of the 57 different serotypes identified, O11:H2 (2 strains), O22:H3 (4 strains), O35:HH11 (2 strains), O52:H3 (2 strains) and O90:H6 (2 strains) were found among isolates from humans and animals (mainly in cats) in Finland and Cuba, and O23:H1a1b (3 strains) among isolates from environmental sources in Slovak Republic and Italy. Most (93–100%) of all strains were susceptible to all anti-microbials tested but resistant (92–96%) to the broad-spectrum penicillins (ampicillin, mezlocillin). No correlation between anti-microbial resistance patterns and serotypes was found.

Beta-lactamase expression in Plesiomonas shigelloides.

We have examined the production of beta-lactamases by 11 clinical and nine environmental isolates of Plesiomonas shigelloides from Czechoslovakia, the Czech Republic and Cuba. Ten isolates (50%) expressed detectable amounts of a chromosomally encoded, non-inducible beta-lactamase, though all isolates showed a broadly similar resistance profile: low-level resistance to ampicillin and higher-level resistance to carbenicillin. All strains were susceptible to cephalosporins and meropenem. Three clinical isolates expressed a beta-lactamase similar to a class 2c carbenicillinase, with a pI of 5.2 and three expressed an enzyme similar to a class 2d oxacillinase, with a pI of 5.3. The environmental isolates produced a variety of penicillinases, indicating that there is a reservoir of heterogeneous beta-lactamase genes in this species.

The Isolation of Plesiomonas shigelloides in Polymicrobial Septicaemia Originating from the Biliary Tree

The first two cases are reported in which Plesiomonas shigelloides was isolated from patients with mixed aerobic/anaerobic septicaemia originating from the biliary tree. The first patient who was previously well survived after removal of a gangrenous gallbladder. The second patient suffered from an extrahepatic cholangiocarcinoma and died of infection thus highlighting the poor prognostic outcome of P. shigelloides septicaemia in immunocompromised patients. Furthermore these cases demonstrate that P. shigelloides can be isolated from mixed infections and may act synergistically with other bacteria to produce disease.

In vitro susceptibilities of Plesiomonas shigelloides to 24 antibiotics and antibiotic-beta-lactamase-inhibitor combinations

The antibiotic susceptibilities of 29 isolates of Plesiomonas shigelloides were studied with 24 antibiotics and antibiotic-inhibitor combinations. Results indicated that all isolates were susceptible to the cephalosporins, penicillins combined with a beta-lactamase inhibitor, aztreonam, and ciprofloxacin. Most isolates were resistant to the penicillins, possibly via production of a penicillinase.

Antimicrobial susceptibility of Plesiomonas shigelloides from patients with diarrhea.

Antimicrobial susceptibility testing was performed on 72 fecal isolates of Plesiomonas shigelloides from patients with diarrhea by using a standard agar dilution technique. All isolates were obtained from a single geographic location. Based on the in vitro activities demonstrated in this study, norfloxacin, ciprofloxacin, and trimethoprim may be effective as oral agents in the treatment of P. shigelloides diarrheal disease. For the parenteral treatment of extraintestinal infection, netilmicin and the cephalosporins had the best in vitro activity.

Isolation of Plesiomonas shigelloides from nutrient broth with brilliant green: its use in screening stool samples from an African population.

The suitability and selectivity of nutrient broth with bile salts and varying concentrations of brilliant green as an enrichment medium was tested for the isolation of Plesiomonas shigelloides from faeces. The broth was most effective at a concentration of 0.001 g/l brilliant green, using faeces deliberately contaminated with Plesiomonas. Two hundred and nineteen faecal samples from Nigeria were then examined for the presence of Plesiomonas and other bacterial pathogens. One yielded Plesiomonas following enrichment in bile salts brilliant green broth (BBG broth) whereas no samples were positive on direct plating.

Isolation of Plesiomonas shigelloides and its distribution in fresh water environments.

A selective medium (KS5) for the isolation of Plesiomonas shigelloides and Aeromonas hydrophila from fresh water fish and water samples was developed by modifying the Rimler-Shotts medium (RS). KS5 medium contains fructose and sucrose as fermentable carbohydrates, and penicillin G and oleandomycin phosphate as selective inhibitors. The mean recovery rates for Plesiomonas and Aeromonas strains on KS5 plates were 93 and 67%, respectively. Plesiomonas strains formed whitish blue or blue colonies while Aeromonas strains produced yellow or yellow orange colonies. On differentiation between Plesiomonas and Aeromonas isolated from fresh water environments, their morphology on KS5 plates were fully consistent with the characteristics of each genus including O/129 sensitivity, starch hydrolysis, and lysine and ornithine decarboxylation. Anaerobic culture was recommended for the selection of Plesiomonas because some reference strains of Pseudomonas could grow on KS5 plates under aerobic conditions. At the drainage outlet of tilapia culture ponds and within about 2km downstream, Plesiomonas shigelloides was isolated on KS5 medium in relatively high densities.

Feasibility of same-day identification of members of the family Vibrionaceae by the API 20E system.

Sixty isolates, comprising nine species of the family Vibrionaceae, were tested with the API 20E 5-h same-day procedure (Analytab Products, Plainview, N.Y.). Included were 27 Aeromonas hydrophila isolates, 10 Aeromonas sobria isolates, 7 Aeromonas caviae isolates, 3 Plesiomonas shigelloides isolates, 3 Vibrio alginolyticus isolates, 3 Vibrio cholerae isolates, 1 Vibrio fluvialis isolate, 5 Vibrio parahaemolyticus isolates, and 1 Vibrio vulnificus isolate. The 5-h profile numbers were specific for the five Vibrio species and the Plesiomonas isolates. The three Aeromonas species shared seven 5-h profile numbers. Of the 63 5-h profile numbers generated by testing each isolate twice, 22 were identical to those found in the overnight analytical profile index. Of these, 20 were correct identifications, and two were incorrect. The remaining 41 5-h profile numbers were not found in the overnight analytical profile index. Because the 5-h analytical profile index does not contain any oxidase-positive organisms, the oxidase value was subtracted from the 63 5-h profile numbers to determine whether misidentifications could occur if the oxidase test was either not performed or not performed correctly. Only five of these factored profile numbers resulted in a possible misidentification. It is feasible, within limitations, to use the 5-h API 20E same-day procedure to identify the more commonly occurring members of the Vibrionaceae. The manufacturer should develop a data base for this purpose.

Inclusion bodies in Plesiomonas shigelloides.

Inclusion bodies were discovered in seven environmental isolates of Plesiomonas shigelloides and the P. shigelloides control (ATCC 14029). Differential staining indicated that the inclusion bodies may be composed of polyphosphates, and developmental stages of the bodies may occur. The inclusion bodies may be useful for rapid presumptive identification of this organism.