Vibrio Parahaemolyticus Bacteria Research Articles

Komparasi Metode Isolasi DNA dalam Mendeteksi Gen toxR Bakteri Vibrio parahaemolyticus pada Udang Vaname (Litopenaeus vannamei)

Vibrio parahaemolyticus bacteria is one of the dangerous pathogens in vaname shrimp farming (Litopenaeus vannamei). Infection with this bacterium can cause mass mortality in shrimp and considerable losses for farmers. This bacterium also has a specific gene, the toxR gene, which is used to detect its presence. The successful detection of this bacterium determines the success of handling the disease it causes. The purpose of the study was to determine the optimal DNA isolation method in detecting the toxR gene of Vibrio parahaemolyticus bacteria in vaname shrimp. The research was conducted by means of molecular biology-based identification using two DNA isolation protocols, namely Chloroform and Boilling lysis buffer. Visualization results showed that toxR gene DNA appeared in samples isolated with Boiling Lysis Buffer in Tryptic Soy Broth (TSB) media culture preparation. This means that the boiling lysis buffer method is a suitable DNA isolation method for detecting the toxR gene of the Vibrio parahaemolyticus bacteria in white shrimp.

ANTIBACTERIAL SCREENING OF SARGASSUM SP EXTRACT. OPPOSING VIBRIO PARAHAEMOLYTICUS BACTERIA

Antibiotics cause resistance to pathogenic bacteria, and the emergence of resistance and bacterial pathogenic infections makes scientists try to find new drugs. One of the efforts is to use marine organisms as a natural antibacterial agent, one of them is Sargassum sp. Sargassum sp contains alginate and iodine which are used in the food, pharmaceutical, cosmetic and textile industries. Apart from that, Sargassum sp. contains active compounds of steroids, alkaloids, phenols and triterpenoids which function as antibacterials, can be used to inhibit the growth of Vibrio sp bacteria which is the cause of firefly disease which is most dangerous for tiger prawns. The purpose of this study was to determine the ability of the extract Sargassum sp. In inhibiting the growth of Vibrio parahaemolyticus bacteria with different levels of solvent start with methanol, ethyl asetat, and n-hexane. The rejuvenation of Vibrio parahaemolyticus and Vibrio harveyimelalai bacteria is carried out in two stages, the first is planting on Triptyc Soy Broth (TSB) medium and the second stage is planting on Triptyc Soy Agar (TSA) medium. Antibacterial sensitivity test using disc diffusion method. The results of the test showed that 1). sargassum sp which is extracted by metanol solvent inhibits the growth of Vibrio parahemolyticus bacteria. 2). sargassum sp which extract by etil sestat and n - heksan solvent can't inhibit the Vibrio parahemolyticus bacteria.

Hydrothermal synthesis of Cu2CoSnS4 nanoparticles: characterization and their applications of electrochemical, antibacterial and photocatalytic performances

Abstract A hydrothermal technique was used to successfully synthesize tetragonal Cu2CoSnS4 (CCTS) nanoparticles and investigate the effect of various thiourea concentrations on structural, morphological and optical properties. XRD analysis revealed the formation of tetragonal CCTS nanoparticles and the average crystallite size (nm) varied from 26 to 40 nm. The Raman studies confirmed the vibrational modes of the CCTS nanoparticles. The FE-SEM images revealed that the thiourea concentrations induced morphological changes in the CCTS nanoparticles, which exhibited the nanosheets changing into spherical structures. TEM images indicated that the CCTS sample had a spherical structure and the SAED pattern demonstrated a polycrystalline nature. The valance states of metallic species, such as Cu+ and Co2+ were further confirmed by XPS. The optical band gap (1.53 eV) was calculated from UV–Visible data and the obtained bandgap value from the literature. The electrochemical measurements of the CCTS TU-10 electrode exhibited pseudocapacitive behavior with a notable specific capacitance of 198 Fg−1 at a scan rate of 10 mV/S along with favorable electrocatalytic activity. In the photocatalytic application, when compared to MB dye (78 %), the CCTS catalyst was found to have a higher degradation efficiency toward CV dye (84 %). For antibacterial studies, the Vibrio parahaemolyticus bacteria exhibited the maximum zone of inhibition at 20 mm for 100 μg/ml. Finally, the experimental results suggested that the synthesized CCTS nanoparticles had better electrochemical, photocatalytic, and antibacterial properties.

ADDITION OF Caulerpa racemosa EXTRACT ON RPS AND CLINICAL SIGNS OF VANAME SHRIMP AFTER Vibrio parahaemolyticus INFECTION

Prevention of AHPND in shrimp can be done by giving immunostimulants, one of which comes fromseaweed extract, explicitly Caulerpa racemosa. This study aimed to determine the effect of the administration of Caulerpa racemosa extract on the relative percent survival (RPS), clinical signs, and behavioral changes of white shrimp after being infected with Vibrio parahaemolyticus. This study used a completely random design with five treatments and three replications, namely K-, K+, and three treatments with the addition of Caulerpa racemosa extract, names P1 (3 μg/g), P2 (6 μg /g), and P3 (9 μg/g). The infection process with Vibrio parahaemolyticus bacteria was carried out 24 hours after being injected with Caulerpa racemosa extract. The results of this study indicate that the highest Relative Percent Survival (RPS) value was P3. Clinical signs of shrimp infected with Vibrio parahaemolyticus were characterized by empty intestines and a whitened hepatopancreas. In addition, there were behavioral changes in shrimp infected with Vibrio parahaemolyticus, namely decreased appetite, passive shrimp movement at the bottom of the aquarium, and erratic swimming movement of shrimp. Based on these results, it could be said that the best treatment is in P3 with a dose of 9 μg/g. This study can provide the best quantity of C. racemosa extract to treat AHPND in shrimp.

Optimisation of the process of acquiring tannin compounds with anti - Vibrio parahaemolyticusfrom the leaves of sea almond tree (Terminalia catappa L.)

Terminalia catappa L. (so-called Bang in Vietnamese) is considered an important medicinal plant in folk medicine for the treatment of digestive disorders and other inflammatory conditions. This plant contains several medicinally essential components, especially the tannin group. This study evaluated a number of factors affecting tannin extraction from the leaves of sea almond tree with the help of response surface methodology (RSM). The factors used to evaluate are solvent concentration, solvent:material ratio (v/w) and soaking time (hours). Research results show that the highest total tannin content (TTC) is 1.54 mg GAE/g raw material, the optimal conditions are determined as follows: ethanol 54%, solvent:material ratio (S:MR) is 41:1 (v/w) and soaking time 2 hours. The experimental values (TTC=1.54±0.02 mg TAE/g raw material) show good agreement with the predicted value (TTC=1.54 mg TAE/g raw material). Experiments evaluating antibacterial activity show that the optimal extract of sea almond tree leaves has the ability to resist Vibrio parahaemolyticus bacteria. The results of this study could pave the way for the mass production of medicinal compounds containing tannins from sea almond tree leaves and improve the usage value of this popular plant.

The Influence of Adding Api-Api Mangrove Leaf Extract as an Immunostimulant on Vaname Shrimp Against Vibriosis Disease Caused by Vibrio parahaemolyticus Bacteria

Vanname shrimp cultivation is one of the most developed aquacultures in Indonesia and makes it one of the commodities with the highest foreign exchange inputs for the Indonesian state from the fisheries sector. Disease attack is one of the main causes of failure in vaname shrimp culture, especially disease attacks caused by Vibrio parahaemolyticus bacteria. Avicennia alba mangrove leaf extract is known to have active compounds that can act as antibacterial. This research was conducted by mixing Avicennia alba mangrove leaf extract with doses of 150 ppm (VA), 250 ppm (VB) and 350 ppm (VC) into commercial feed by re-pelletting. Positive control (KV+) treatment used commercial vitamins added to the feed. The vaname shrimp used was 60 days old with an average initial weight of 8.9 grams. The aquariums used were 15 pieces with a size of 60 x 30 x 40 cm and a volume of 54 liters of water. The white shrimp challenge test with Vibrio parahaemolyticus bacteria with a concentration of 106 Cfu/ml as much as 0.2 ml. The clinical symptoms of white vaname shrimp include pale white hepatopancreas, blackened tail, grippy swimming legs, necrosis in some parts of the body, necrosis of the tail, blackened and necrotic rectum, broken intestines that are partially brown and starting to turn white, empty intestines, presence of white feces. appears in the aquarium, decreased appetite, as well as not actively swimming. The results showed that Avicennia alba leaf extract had an effect on absolute weight growth (W), growth rate (GR), specific growth rate (SGR), survival rate (SR), total haemocyte count (THC) and phagocytic activity of white shrimp. The dose of 350 ppm VC treatment was the most influential dose as an immunostimulant in this study.

The Effectiveness of Lactic Acid Bacteria on the Immune System of Vannamei Shrimp Infected with Bacteria Vibrio Parahaemolyticus

Diseases are a major obstacle in shrimp farming because they can cause relatively high mortality and decrease the quality of the aquaculture environment. One dangerous shrimp disease is caused by Vibrio bacteria. Today, people are turning to biological control methods by utilizing lactic acid bacteria that live as microflora in the digestive tract of aquaculture animals. This study aimed to analyze the potential of lactic acid bacteria isolates from the gut of vannamei shrimp as probiotic candidates and to analyze selected lactic acid bacteria isolates that were effective in increasing the immune response and survival of vannamei shrimp infected with Vibrio parahaemolyticus bacteria. Four BAL isolates were isolated, designated as 8A, 11B, F1, and G2. BAL isolates showed antibacterial activity against Vibrio parahaemolyticus bacteria with an inhibition zone of up to 11.5 mm, capable of living at acidic to alkaline pH (1.5-7.2), bile salts (3000 ppm) and negative catalase. The characterization of several tests such as gram-positive, inhibition test and sugar fermentation test and comparison of LAB characterization showed that LAB isolates (G2) from the intestine of vannamei shrimp were classified as Pediococcus acidilactici. The application of probiotics, namely LAB (G2) to vannamei shrimp through feed can stimulate the vaname shrimp immune system after being challenged with V. parahaemolyticus, as indicated by an increase in the number of hemocytes (THC), phagocytosis activity, and suppressing the population of V. parahaemolyticus in vannamei shrimp.

Activity of Garlic (Allium Sativum L.) Extract Against Vibrio Parahaemolyticus Bacteria

Vibriosis is the primary bacterial illness shrimp farmers face due to mass death in young shrimp, called Early Mortality Syndrome (EMS). The etiology of this disease is Vibrio parahaemolyticus bacterium. Garlic (Allium sativum L.) is one of the natural compounds that can suppress the growth of Vibrio parahaemolyticus bacteria. This study aims to assess the effect of garlic extract on the growth of Vibrio parahemolyticus bacteria in 96 percent ethanol solvent at garlic extract concentrations of 20%, 40%, 60%, and 80%. The Kirby-Bauer disc diffusion method is utilized, which estimates the diameter of the inhibitory zone surrounding the disc. Making garlic extract by maceration using 96% ethanol solvent and evaporated with a rotary vacuum evaporator. The results showed that a ethanol 96% extract of garlic with concentrations of 20%, 40%, 60%, and 80% had antibacterial activity on V. parahaemolyticus bacteria with an average inhibition zone diameter of 4.9 mm, 8.4 mm, 9.4 mm, 11.9 mm, respectively. This study concluded that garlic extracts have antibacterial activity against Vibrio parahaemolyticus.

LptD-antigen system on gold nanoparticles: an innovative strategy in the nanovaccine development

Nanovaccine development is a growing research field in which the development of new carriers and bioconjugation approaches is a priority. In this sense, this report describes for the first time, the development of a novel conjugate that consists of gold nanoparticles (AuNPs) obtained by a one-step synthesis using an immunogenic peptide of the Lipopolysaccharide-assembly protein LptD from Vibrio parahaemolyticus bacteria as a reducing and capping agent. The resulting LptD@AuNPs compounds were fully characterized and the results showed the high capacity of the peptide to form complexes and reduce gold ions. The reaction yield estimated was higher than 83% and the chemical integrity of the peptide on the NP surface revealed a tyrosine amino acid bonding on the AuNP surface. Furthermore, the LptD@AuNP system showed high colloidal stability in a wide pH range (3–11 pH values), where the hydrodynamic diameter and Zeta potential behavior were strongly influenced by the functional groups of the antigenic peptide. The cytotoxicity assays showed that the obtained system is safe for mouse leukocytes, while immunized mice with LptD@AuNPs produced specific IgG antibodies. These encouraging results revealed the efficacy of some antigenic peptides as reducers and capping agents, in addition, opening the path to determine immunogenicity and immunoprotective efficacy of the LptD@AuNP system against the disease induced by Vibrio parahaemolyticus.

The correlation between ectoparasite infestation and total Vibrio parahaemolyticus bacteria in Pacific white shrimp (Litopenaeus vannamei) in Super Intensive Ponds

This study aimed to determine the type, intensity, degree of ectoparasite infestation and its correlation to the total V. parahaemolyticus bacteria in super-intensive pacific white shrimp culture. In this study, sampling was carried out in 3 super-intensive pacific white shrimp pond areas spread out in East Java, namely Bangil, Tuban, and Lamongan with 50 shrimps (PL30-PL 40). The obtained data underwent regression and correlation analysis. Based on the results, there were three types of ectoparasites, namely Zoothamnium sp., Epistylis sp. and Vorticella sp. High ectoparasite intensity was found in pacific white shrimp from Lamongan and Tuban ponds, namely 76 and 55 individuals/shrimp, respectively, showing the heavy infestation. High total V. parahaemolyticus bacteria was found in Tuban (1.16 × 105 CFU/gr) and Lamongan (1.16 × 105 CFU / gr) ponds. Based on the results, the coefficient value was R = 0.807 showing positive correlation of V. parahaemolyticus with the increasing parasite intensity and low oxygen levels.

Janus Micromotors Coated with 2D Nanomaterials as Dynamic Interfaces for (Bio)-Sensing.

In this work, we study the interaction of graphdiyne oxide (GDYO)-, graphene oxide (GO)-, or black phosphorous (BP)-wrapped Janus micromotors using a model system relying on a fluorescence-labeled affinity peptide, which is released upon specific interaction with a target Cholera Toxin B. Such ON-OFF-ON system allows mimicking similar processes occurring at (bio)-interfaces and to study the related sorption and desorption kinetics. The distinct surface properties of each nanomaterial play a critical role in the loading/release capacity of the peptide, greatly influencing the release profiles. Sorption obeys a second-order kinetic model using the two-dimensional (2D) nanomaterials in connection with micromotors, indicating a strong influence of chemisorption process for BP micromotors. Yet, release kinetics are faster for GDYO and GO nanomaterials, indicating a contribution of π and hydrophobic interactions in the probe sorption (Cholera Toxin B affinity peptide) and target probe release (in the presence of Cholera Toxin B). Micromotor movement also plays a critical role in such processes, allowing for efficient operation in low raw sample volumes, where the high protein content can diminish probe loading/release, affecting the overall performance. The loading/release capacity and feasibility of the (bio)-sensing protocol are illustrated in Vibrio cholerae and Vibrio parahaemolyticus bacteria cultures as realistic domains. The new concept described here holds considerable promise to understand the interaction of micromotor with biological counterparts in a myriad of biomedical and other practical applications, including the design of novel micromotor-based sensors.

Saccharification of macroalgal polysaccharides through prioritized cellulase producing bacteria

Marine macroalgal cell wall is predominantly comprised of cellulose (polysaccharide) with the complex chain of glycosidic linkages. Bioethanol production from macroalgae entails breaking this complex chain into simple glucose molecule, which has been the major challenge faced by the industries. Cellulases have been preferred for hydrolysis of cellulose due to the absence of inhibitors affecting the subsequent fermentation process. Cellulose degrading bacteria were isolated from wide-ranging sources from marine habitats to herbivore residues and gastrointestinal region. The investigation reveals that Vibrio parahaemolyticus bacteria has higher hydrolytic capacity with salt tolerance up to 14% and 3.5% salinity is optimum for growth. Higher hydrolytic activity of 2.45 was recorded on carboxymethyl cellulose medium at 48 h and hydrolytic activity of 2.46 on Ulva intestinalis hydrolysate, 3.06 on Ulva lactuca hydrolysate at 72 h of incubation. Total activity of enzyme of 2.11 U/ml and specific activity of 6.05 U/mg were recorded at 24 h. Enzyme hydrolysis of macroalgal biomass; U. intestinalis and U. lactuca produced 135.9 mg/g and 107.6 mg/g of reducing sugar respectively. The study reveals that the enzyme extracted from salt tolerant Vibrio parahaemolyticus bacteria is suitable for optimal saccharification of seaweed polysaccharides towards biofuel production.

Ginger and its component shogaol inhibit Vibrio biofilm formation in vitro and orally protect shrimp against acute hepatopancreatic necrosis disease (AHPND)

Specific isolates of Vibrio parahaemolyticus bacteria that cause acute hepatopancreatic necrosis disease (AHPND) colonize and form biofilms on the chitin lining of the shrimp stomach where they release toxins that cause necrosis in the adjacent hepatopancreas. One potential strategy to prevent AHPND is to inhibit biofilm formation in the stomach using feed additives. Thus, we tested ethanolic extracts of ginger (0.2 mg/ml and 2 mg/ml) for their ability to inhibit biofilm formation by an AHPND-causing isolate of V. parahaemolyticus (3HP) using chitosan coated 96-well polystyrene plastic plates to mimic the shrimp stomach lining. The extract significantly (p < .05) inhibited biofilm formation in those plates but did not inhibit cell growth in broth culture in parallel uncoated plates. Subsequent in vivo immersion-challenge experiments using 3HP (106 CFU/ml) showed that use of extract-supplemented feed (0.2 mg/g and 2 mg/g) for 7 days prior to challenge protected whiteleg shrimp (Penaeus vannamei) against mortality (40%–60% higher survival) when compared to control shrimp fed un-supplemented feed. Extract supplementation had no apparent negative effect on feed palatability or on shrimp growth. Three potentially bioactive compounds known to occur in ginger extracts (6-gingerol, 8-gingerol, and 6-shogaol) were also tested for efficacy in biofilm inhibition in vitro, and 6-shogaol was found to be the most potent followed by the others in descending order. None were found to affect bacterial growth in broth. The results of this study suggested that the use of ginger-based or other feed additives that inhibit biofilm formation may constitute a practical approach to reduce the negative impact of AHPND in shrimp aquaculture.

Detection of tlh and tdh genes in Vibrio Parahaemolyticus inhabiting farmed water ecosystem used for L. Vannamei aquaculture

Vibrio parahaemolyticus is hallophilic gram-negative bacteria that live as natural inhabitant in aquatic environment. All Vibrio parahaemolyticus strain known to have thermolabile hemolysin encoded by tlh gene as species marker. Thermostable direct hemolysin encoded by tdh gene is responsible for regulating virulence factor in Vibrio parhaemolyticus. Aim of this research is to detect tlh and tdh gene from water of L. vannamei aquaculture in Rembang regency. Colonies of green-blueish bacteria grew from isolation of L. vannamei aquaculture water in CD-VP media which was identified as Vibrio parahaemolyticus. Colonies of V. parahaemolyticus grew to be small and green-blueish bacteria colonies in TCBS agar. Result of molecular analysis showed that bacteria isolated from water sample are specifically identified as Vibrio parahaemolyticus bacteria by the detection of tlh gene. Vibrio parahaemolyticus isolated from water of L. vannamei aquaculture detected as tdh negative that indicates tdh gene is not present in isolated bacteria. Vibrio parahaemolyticus isolate were cultured in Wagatsuma agar for tdh gene confirmation test that showed Kanagawa negative result, which indicated that V. parahaemolyticus did not produce thermostable direct hemolysin. These results showed that Vibrio parahaemolyticus isolated from aquatic environment of L. vannamei aquaculture in Rembang regency did not show virulence factors.

Seasonal variation in abundance of total and pathogenic Vibrio parahaemolyticus bacteria in oysters along the southwest coast of India.

The seasonal abundance of Vibrio parahaemolyticus in oysters from two estuaries along the southwest coast of India was studied by colony hybridization using nonradioactive labeled oligonucleotide probes. The density of total V. parahaemolyticus bacteria was determined using a probe binding to the tlh (thermolabile hemolysin) gene, and the density of pathogenic V. parahaemolyticus bacteria was determined by using a probe binding to the tdh (thermostable direct hemolysin) gene. Furthermore, the prevalence of V. parahaemolyticus was studied by PCR amplification of the toxR, tdh, and trh genes. PCR was performed directly with oyster homogenates and also following enrichment in alkaline peptone water for 6 and 18 h. V. parahaemolyticus was detected in 93.87% of the samples, and the densities ranged from <10 to 10(4) organisms per g. Pathogenic V. parahaemolyticus could be detected in 5 of 49 samples (10.2%) by colony hybridization using the tdh probe and in 3 of 49 samples (6.1%) by PCR. Isolates from one of the samples belonged to the pandemic serotype O3:K6. Twenty-nine of the 49 samples analyzed (59.3%) were positive as determined by PCR for the presence of the trh gene in the enrichment broth media. trh-positive V. parahaemolyticus was frequently found in oysters from India.