Molecular quantitation and characterization of Vibrio cholerae from different seafood obtained from wetmarket and supermarket

Vibrio cholerae still represents a significant threat to human health worldwide despite the advances in hygiene, consumer knowledge, food treatment and food processing. In Malaysia, statistics in year 2009 have shown that among the food and water borne diseases, food poisoning has the highest incidence rate of 36.17 per 100,000 populations and with a mortality rate of 0.01 per 100,000 populations. In this study, 22 seafood samples comprising of fish, squid, crustacean and mollusks purchased from wet market and supermarket were analyzed. The Most Probable Number (MPN) and real time PCR was used to enumerate the Vibrio cholerae in seafood sample. The results showed that MPN-real time PCR of the samples from wet market had a maximum of > 1100 MPN/g compare to 93 MPN/g enumerated from the MPN plate. The MPN-real time PCR in the samples from supermarket indicated 290 MPN/g as compared to 240 MPN/g enumerated from the MPN plate. The standard curves showed that there was a good linear correlation between the Ct values. The minimum level of detection of Vibrio cholerae standard DNA at targeted gene was 3 × 10 -5 ng/μl

Editorial : novel approaches to the treatment of multidrug-resistant bacteria, Volume II

No abstract available.http://www.frontiersin.org/Pharmacologyam2023Paraclinical Science

Editorial : Novel approaches to the treatment of multidrug-resistant bacteria

No abstract available.https://www.frontiersin.org/journals/pharmacologydm2022Paraclinical Science

Vibrio vulnificus: An Environmental and Clinical Burden

Vibrio vulnificus is a Gram negative, rod shaped bacterium that belongs to the family Vibrionaceae. It is a deadly, opportunistic human pathogen which is responsible for the majority of seafood-associated deaths worldwide. V. vulnificus infection can be fatal as it may cause severe wound infections potentially requiring amputation or lead to sepsis in susceptible individuals. Treatment is increasingly challenging as V. vulnificus has begun to develop resistance against certain antibiotics due to their indiscriminate use. This article aims to provide insight into the antibiotic resistance of V. vulnificus in different parts of the world as well as an overall review of its clinical manifestations, treatment, and prevention. Understanding the organism's antibiotic resistance profile is vital in order to select appropriate treatment and initiate appropriate prevention measures to treat and control V. vulnificus infections, which should eventually help lower the mortality rate associated with this pathogen worldwide

Prevalence and characterisation of antibiotic resistance of Vibrio parahaemolyticus from seafood in Selangor, Malaysia / Vengadesh Letchumanan

Aquaculture industry has been professed as one of the fast-growing industries that serves a major source of seafood and revenue to many countries worldwide. Despite the nutritional benefits of seafood consumption, health risks linked to seafood consumption cannot be disregarded. Microbiological safety of seafood is of global concern recent years due to occurrence of seafood-borne cases and increase reports on antibiotic resistance among Vibrio parahaemolyticus isolated from seafood. The emergence of antimicrobial resistant V. parahaemolyticus poses treat to human health. In regard to increase reports on V. parahaemolyticus as a causative agent of seafood-borne illness, the study aimed to enumerate and characterise the antibiotic resistance profiles of V. parahaemolyticus isolated from seafood. A total of 770 seafood samples namely shrimp and shellfish were collected from both local wetmarket and supermarket in Selangor. The enumeration and identification using microbiological plating method on selective agar, thiosulphate citrate bile salt sucrose (TCBS) agar revealed that all seafood samples collected from wetmarket and supermarket sites were contaminated with Vibrio sp. The seafood samples analyzed had a microbial load of 2.29 log CFU/g to 6.63 log CFU/g. The toxR-PCR assay identified positive amplification of toxR gene in 50% (385/770) of the presumptive isolates. 32/385 (8.3%) isolates harboured the thermostable-related direct haemolysin (trh) gene and none with thermostable direct hemolysin (tdh) gene. The antibiotic susceptibility test revealed a total of 102 different types of antibiograms profiles among the V. parahaemolyticus isolates. The isolates were seen to be resistant to at least one type of antibiotic tested with MAR index ranged from 0 to 0.79. The chloramphenicol (catA2) gene was detected in 18/22 chloramphenicol-resistant isolates and 18/193 kanamycin-resistant isolate was positive for kanamycin aphA-3 gene. Further analysis on the plasmid profiles of V. parahaemolyticus isolates revealed 1-7 plasmids, with sizes ranging from 1.2kb to 10kb. There was no correlation seen between the plasmid profiles and antibiotic resistance patterns. Even within the isolates with same resistance profiles, the plasmid profiles were different and a few isolates even did not exhibit any plasmids. The isolates either demonstrated plasmidial or chromosomally mediated antibiotic resistance after plasmid curing assay. In conclusion, the results demonstrate that all the seafood samples collected are contaminated with V. parahaemolyticus regardless the sampling location and some of which carried the trh-gene which are potential to cause foodborne illness. The occurrence of multidrug resistance emphasizes the importance of study of antibiotic susceptibility of V. parahaemolyticus. Hence, constant monitoring of the prevalence and characterisation of resistance profiles of V. parahaemolyticus is needed to ensure food safety and human wellbeing

Detection of Vibrio Cholerae and Vibrio parahaemolyticus in seafood using MPN-PCR based technique, and their molecular characteristics

Seafood is professed by consumers worldwide to be healthy and nutritious food due to abundance of scientific and documented health benefits. Approximately 90% of global aquaculture production is based in Asia. Nevertheless, recent food borne outbreaks are closely associated with seafood consumption. Seafood is known as a vehicle of transmission of food borne bacteria and causes human illness worldwide. In Malaysia, statistics in year 2009 have shown among the food and water borne diseases, food poisoning has the highest incidence rate of 36.17 per 100,000 populations and with a mortality rate of 0.01 per 100,000 populations. The purpose of this study is to apply and enumerate Vibrio cholerae and Vibrio parahaemolyticus from seafood samples by utilizing the Most Probable Number Method (MPN) and several molecular typing methods including polymerase chain reaction (PCR), enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) and plasmid profiling. The application of conventional method using the Most Probable Number (MPN) method with selective enrichment broth and agar medium is very useful in isolating Vibrio cholerae and Vibrio parahaemolyticus. This method is coupled with PCR based method to obtain specific, sensitive and precise results. The densities enumerated by the MPN-Real Time PCR targeting epsM gene and the MPN-PCR targeting toxR gene is higher than those by MPN-Plate. Genomic DNA of 104 Vibrio cholerae isolates was confirmed by a specific optimized multiplex PCR program targeting hem gene, hlyA gene, ctx gene and zot gene. 10 isolates were tested positive for cholera toxin gene (ctx) gene. All the isolates were positive to hem gene at 519bp and hlyA gene at 738bp but none was tested positive for zot gene. On the other hand, all the 100 Vibrio parahaemolyticus isolates were tested positive for regulatory gene, toxR yielded 368bp. The PCR amplification of the respective genes is a rapid and reliable method of detecting Vibrio cholerae and Vibrio parahaemolyticus isolates from seafood samples. ERIC sequences are short, highly conserved 126 bp non-coding regions found in the Enterobacteriaceae. Its location in bacterial genomes allows discrimination at the genus, species and serovars levels. Dendrogram of ERIC-PCR were analyzed using the Bionumerics Version 6.0 (Applied Maths, Germany) software. From 104 isolates of Vibrio cholerae, ERIC-PCR with primers ERIC-1 and ERIC-2 produced 15 clusters and 9 single isolates at 40% similarity. Where else, 100 isolates of Vibrio parahaemolyticus produced 15 clusters and 6 single isolates at 40% similarity. The results demonstrated that ERIC-PCR is a excellent tool for differentiation and characterization of Vibrio species. Plasmids of Vibrio cholerae and Vibrio parahaemolyticus vary in size from 2.2 Kb to more than 7.4 Kb. Despite limited knowledge on their function, their presence is frequently used for strain differentiation in epidemiological studies. Plasmid profiling of 104 Vibrio cholerae isolates clustered into 19 groups based on the number and pattern of the bands. Where else, plasmid profiling of 100 Vibrio parahaemolyticus isolates clustered into 11 groups based on the number and pattern of the bands. As a conclusion, the concern about possible health illness from Vibrio species, especially when seafood remains as a vehicle of transmission of Vibrio, will continue into the likely future. Therefore, to establish effective control measures to reduce the risk of this bacterium infection and to ensure the safety of foods, surveillance and epidemiology, the employment of molecular methods for the detection of Vibrio cholerae and Vibrio parahaemolyticus in food and environment is important. The results from this study could serve as vital information in Vibrio spp. epidemiology, surveillance, better infection control measures and support of public health policy

A revolutionizing approach to autism spectrum disorder using the microbiome

The study of human microbiota and health has emerged as one of the ubiquitous research pursuits in recent decades which certainly warrants the attention of both researchers and clinicians. Many health conditions have been linked to the gut microbiota which is the largest reservoir of microbes in the human body. Autism spectrum disorder (ASD) is one of the neurodevelopmental disorders which has been extensively explored in relation to gut microbiome. The utilization of microbial knowledge promises a more integrative perspective in understanding this disorder, albeit being an emerging field in research. More interestingly, oral and vaginal microbiomes, indicating possible maternal influence, have equally drawn the attention of researchers to study their potential roles in the etiopathology of ASD. Therefore, this review attempts to integrate the knowledge of microbiome and its significance in relation to ASD including the hypothetical aetiology of ASD and its commonly associated comorbidities. The microbiota-based interventions including diet, prebiotics, probiotics, antibiotics, and faecal microbial transplant (FMT) have also been explored in relation to ASD. Of these, diet and probiotics are seemingly promising breakthrough interventions in the context of ASD for lesser known side effects, feasibility and easier administration, although more studies are needed to ascertain the actual clinical efficacy of these interventions. The existing knowledge and research gaps call for a more expanded and resolute research efforts in establishing the relationship between autism and microbiomes