Case Report: Vibrio fluvialis isolated from a wound infection after a piercing trauma in the Baltic Sea

Vibrio spp. are Gram-negative bacteria found in marine ecosystems. Non-cholera Vibrio spp. can cause gastrointestinal infections and can also lead to wound infections through exposure to contaminated seawater. Vibrio infections are increasingly documented from the Baltic Sea due to extended warm weather periods. We describe the first isolation of Vibrio fluvialis from a wound infection acquired by an impalement injury in the shallow waters of the Baltic Sea. The severe infection required amputation of the third toe. Whole genome sequencing of the isolate was performed and revealed a genome consisting of two circular chromosomes with a size of 1.57 and 3.24 Mb.

Virulence and Antibiotic Resistance Characteristics of Vibrio Isolates From Rustic Environmental Freshwaters

The study investigated the occurrence of antimicrobial resistance genes and virulence determinants in Vibrio species recovered from different freshwater sheds in rustic milieu. A total of 118 Vibrio isolates comprising Vibrio fluvialis (n=41), Vibrio mimicus (n=40) and V. vulnificus (n=37) was identified by amplification of ToxR, vmh and hsp60 genes. The amplification of virulence genes indicated that V. mimicus (toxR, zot, ctx, VPI, and ompU) genes were detected in 12.5%, 32.5%, 45%, 37.5% and 10% respectively. V. fluvialis genes (stn, hupO and vfh) were harboured in 48.8%, 14.6% and 19.5% isolates congruently. The other virulence genes that include vcgC and vcgE were observed in 63.1% and 29% of isolates belonging to V. vulnificus. With the exceptions of imipenem, meropenem and ciprofloxacin, most isolates exhibited more than 50% resistance to antibiotics. The antimicrobial resistance was more prevalent for polymyxin B (100%), azithromycin (100%) and least in ciprofloxacin (16.1%). Multiple antibiotic resistance index range was 0.3 and 0.8 with most isolates showing MARI of 0.8. The blaTEM, AmpC, blaGES, blaIMP, blaOXA-48 and blaKPC genes were detected in 53.3%, 42%, 29.6%, 16.6%, 15%, 11.3% and 5.6% of the isolates. Non-beta lactamases such as streptomycin resistance (aadA and strA), gentamicin resistance (aphA1) and quinolone resistance gene (qnrVC) were found in 5.2%, 44.3%, 26% and 2.8%. Chloramphenicol resistance genes (cmlA1 and catII) were found in 5.2% and 44.3% among the isolates. Our findings reveal the presence of antimicrobial resistance genes and virulent Vibrio species in aquatic environment which can have potential risk to human and animal’s health.

Chiral Synthesis of 3-Amino-1-phenylbutane by a Multi-Enzymatic Cascade System

Asymmetric synthesis of chiral amines from prochiral ketones using transaminases is an attractive biocatalytic strategy. Nevertheless, it is hampered by its unfavorable thermodynamic equilibrium. In the present work, an insitu by-product removal strategy was applied for the synthesis of 3-amino-1-phenylbutane (3-APB) by coupling a transaminase with a pyruvate decarboxylase (PDC), which does not require the use of any expensive additional cofactor. Using this strategy, the pyruvate obtained in the transamination reaction is transformed by PDC into acetaldehyde and CO2 which are of high volatility. Two different transaminases from Chromobacterium violaceum (CviTA) and Vibrio fluvialis (VflTA) were characterized to find out the appropriate pH conditions. In both cases, the addition of PDC dramatically enhanced 3-APB synthesis. Afterwards, different reaction conditions were tested to improve reaction conversion and yield. It was concluded that 30 °C and a 20-fold alanine excess lead to the best process metrics. Under the mentioned conditions, yields higher than 60% were reached with nearly 90% selectivity using both CviTA and VflTA. Moreover, high stereoselectivity for (S)-3-APB was obtained and ee of around 90% was achieved in both cases. For the first time, the asymmetric synthesis of 3-APB using PDC as by-product removal system using CviTA is reported.

Vibrio fluvialis – Unusual Case of Cellulitis Leading to Sepsis

Vibrio fluvialis, an enteric, Gram negative bacterium commonly isolated from sewage/ sea water contaminated with human and animal’s faeces. Infections with this unusual organism can cause cholera like bloody diarrhoea and also wound infection. Vibrio fluvialis causing skin infection and sepsis is uncommon with very few cases reported worldwide. It is an emerging pathogen with distinct features as compares to the other species of Vibrio group of bacteria in terms of high virulence and adaptability in hostile environments. Here is a case report of skin and soft tissue infection by Vibrio fluvialis, in a patient suffering from severe pedal oedema due to nephrotic syndrome and right lower limb deep vein thrombosis . Case Report: A young male, farmer, presented with ascites, pedal oedema, puffy face due to nephrotic syndrome.He developed skin and soft tissue infection resulted from skin atrophy and ulcer due to pedal oedema as well as right limb deep vein thrombosis . He had high grade fever, leucocytosis, anaemia, hypoproteinaemia, right leg cellulites and features of sepsis requiring wound debridement. Immunodeficiency and corticosteroids associated immunosuppression were absent in this patient. He was diagnosed with nephrotic syndrome due to minimal change disease. Conclusion: Vibrio fluvialis infection can occur due to wound contaminated with sewage water leading to skin and soft tissue infection and life threatening sepsis.

Antibacterial activity of lupeol from the bark of Dehaasia cuneate (Lauraceae)

Dehaasia is a member Lauraceae. It is locally known as ‘gajus hutan’ or ‘pekan’. A triterpenoid, lupeol was isolated from the bark of Dehaasia cuneate. The structure of the isolated compound was determined using spectroscopic methods, such as UV–vis, FT-IR, 1D and 2D NMR, and mass spectrometer. The isolated compound was tested against Gram-negative and positive bacteria using agar disc diffusion technique. The results showed that lupeol had a moderate inhibition zone value of 10.0±0.00 mm against Gram-negative Serratia marcescens ATCC 14756 whereas low inhibition which is 7.0±0.00 mm against Escherichia coli ATCC 25922, Vibrio fluvialis ATCC 33809, Bacillus subtilis ATCC 6633, and Methicillin-resistant Staphylococcus aureus (MRSA) ATCC 43300.

Antibiogram signatures of non-cholera causing Vibrio species recovered from environmental niches in Eastern Cape, South Africa.

Background: The use of antibiotics globally has helped reduce mortality and morbidity rate due to its ability to effectively treat bacterial infections in both humans and animals. However, the menace of antimicrobial resistance has become a challenge to public health due to its increased mortality and morbidity rate. This study determined the antibiogram pattern of non-cholera causing Vibrio species against a panel of 11 antibiotics that are wildly used for treatment. Multiple antibiotic resistance phenotype, multiple antibiotic resistant indices and minimum inhibitory concentration (MIC) of test antibiotics were also determined.Results: Polymerase chain reaction (PCR) was used to confirm 100 isolates of Vibrio parahaemolyticus, 82 and 46 isolates of Vibrio vulnificus and Vibrio fluvialis respectively, collected from the culture collections of the Applied and Environmental Microbiology Research Group (AEMREG), University of Fort Hare. Thereafter, disc diffusion method was used to determine the antibiogram pattern of target non-cholera causing Vibrio species against a panel of 11 antibiotics that are of clinical importance. The highest rate of Vibrio parahaemolyticus resistance was observed against tetracycline (22 %) and nalidixic acid (16 %). Vibrio fluvialis also displayed highest rate of resistance against tetracycline (28 %) and nalidixic acid (28 %), while Vibrio vulnificus isolates exhibited highest rate resistance against imipenem (40 %) and tetracycline (22 %). A total of 38 MARP patterns were observed and the MAR indices ranged between 0.3 and 0.8. Against the resistant Vibrio parahaemolyticus and Vibrio fluvialis isolates, minimum inhibitory concentration ranged from 16 µg/ml to 2048 µg/ml for both tetracycline and nalidixic acid, while against Vibrio vulnificus isolates, minimum inhibitory concentration ranged from 8 µg/ml to 256 µg/ml for both imipenem and nalidixic acid. Conclusions: Results obtained from this study is an indication that antibiotic resistant bacteria that could pose as threat to health of humans and animals are present in the environment.

Functional insights of two MATE transporters from Vibrio fluvialis

SummaryFunctional characterization of H- and D-MATE (Multidrug and Toxin Extrusion) transporters from clinical isolates of Vibrio fluvialis revealed H-type conferred resistance to fluoroquinolones, ethidium bromide and safranin whereas D-type exhibited marginal resistance towards ethidium bromide only. Both H-/D-type transporters were inhibited by reserpine resulting in increased intracellular norfloxacin concentration. The efflux was facilitated by both Na+/K+ ions, suggesting that these efflux pumps were ion-dependent. In presence of various classes of EPIs, there was decrease in MIC exhibited by H-/D-type efflux pumps towards norfloxacin which didn’t translate into transport inhibition. But reserpine presented a conclusive pattern with decrease in MIC towards norfloxacin and increased norfloxacin accumulation inferring maximum inhibition. Substrate binding and electrostatic charge distribution of both the transporters was similar to other known MATE transporters. The H-type exhibited 10 transmembranes and D-type exhibited 11 TMs which was different from other MATE transporters known to have 12 TMs (Transmembranes). Data derived from molecular docking and ion binding studies revealed that Aspartic Acid residue in 1st TM acts as ion binding site with transport mechanism similar to NorM. Electrostatic potential map of both the transporters revealed that there is a cavity formation within the transporters surrounded by charged electronegative amino acid residues. Interestingly, surface models of both transporters revealed that 1st TM forms covalent bond with 7th TM towards extracellular space. Docking studies also revealed that reserpine covalently binds to central pocket of both transporters and serves as excellent EPI against these transporters as evidenced by MIC and drug accumulation assays.