scholarly journals Effects of salinity and shaking condition on the growth and virulence of Vibrio parahaemolyticus

2020 ◽  
Vol 18 (2) ◽  
pp. 349-362
Author(s):  
Tran Ngoc My Hanh ◽  
Tran Van Nhi ◽  
Nguyen Thi Thu Hoai
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Extracellular Enzymes ◽  
Brain Heart Infusion ◽  
Halophilic Bacterium ◽  
Chitinase Activity ◽  
Early Mortality Syndrome ◽  
Potential Pathogen ◽  
Salinity Increase ◽  
The One ◽  
Acute Hepatopancreatic Necrosis Disease

Vibrio parahaemolyticus is a Gram-negative halophilic bacterium that is found in estuarine, marine and coastal environment. This organism is the major causative agent of Early Mortality Syndrome (EMS) or Acute Hepatopancreatic Necrosis Disease (AHPND) which resulted in serious damages to cultured shrimp industry. Understanding the effect of environmental factors on the growth and virulence of this potential pathogen would be beneficial for preventing its outbreak. In this study, the growth and virulence of V. parahaemolyticus was examined under different salinity and shaking condition. V. parahaemolyticus XN9 was cultured in Brain Heart Infusion (BHI) medium with different sodium chloride concentrations (2.0, 2.5 and 3.0%) and different shaking conditions (0, 120 and 240 rpm). The growth of the bacterium was recorded over 8h and six extracellular enzymes of V. parahaemolyticus XN9 including caseinase, hemolysin, lecithinase, lipase, gelatinase, chitinase were investigated using agar-based method. The growth of V. parahaemolyticus was varied among different salinity and shaking conditions. It showed the best growth at 2.0% NaCl and 240 rpm. No change in the enzymatic activity (EA) of the tested extracellular enzymes was observed while changing salinity except the significant decline of gelatinase from 3.49±0.19 to 2.77±0.17 mm following salinity increase (p < 0.05). On the other hand, regarding shaking condition, lipase was the one to increase its activity significantly following the increase of shaking speed (p < 0.05). While caseinase, lecithinase, gelatinase and lipase were well expressed in V. parahaemolyticus, no hemolytic and chitinase activity was observed in any tested conditions. In summary, our study showed that 2.0 % NaCl and 240 rpm shaking promoted the best growth of V. parahaemolyticus and resulted in highest activity of gelatinase and lipase in this bacterium.

scholarly journals Effects of different shrimp extracts on Vibrio parahaemolyticus growth and virulence

2021 ◽  
Vol 63 (4) ◽  
pp. 62-65
Author(s):  
Nguyen Thuan Thien Truong ◽  
◽  
The Hao Nguyen ◽  
Cong Chinh Bui ◽  
Thi Thu Hoai Nguyen ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Vibrio Parahaemolyticus ◽  
Extracellular Enzymes ◽  
Brain Heart Infusion ◽  
Growth Curves ◽  
Bacterial Virulence ◽  
Counting Method ◽  
Tiger Prawn ◽  
Extracellular Enzymatic Activities ◽  
Acute Hepatopancreatic Necrosis Disease

Vibrio parahaemolyticus is the main causative agent of acute hepatopancreatic necrosis disease (AHPND) in shrimp. This study aimed to investigate how shrimp extracts affect the growth and virulence of an AHPND-causative strain known as V. parahaemolyticus XN9. To this end, the bacteria was cultured in media containing 3% extract of each of five shrimp types and their growth kinetics were compared against that from bacteria grown in brain-heart infusion (BHI) media. Eight-hour growth curves were constructed using the plate-counting method. The activity of five extracellular enzymes that contribute to bacterial virulence was examined using the agar-based method. The results showed that V. parahaemolyticus XN9’s growth was strongly enhanced in all five shrimp extract media with the highest increase (25% greater than the BHI medium) found in the giant tiger prawn extract. Additionally, all the shrimp extracts boosted the extracellular enzymatic activity of V. parahaemolyticus XN9, although to different extents. In summary, the shrimp extracts, particularly that from the prawns, not only promoted the viability and growth of V. parahaemolyticus XN9 but also its extracellular enzymatic activities.

scholarly journals Draft Genome Sequence of Vibrio owensii Strain SH-14, Which Causes Shrimp Acute Hepatopancreatic Necrosis Disease

2015 ◽  
Vol 3 (6) ◽  
Author(s):  
Liyuan Liu ◽  
Jinzhou Xiao ◽  
Xiaoming Xia ◽  
Yingjie Pan ◽  
Shuling Yan ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Genome Sequence ◽  
Vibrio Parahaemolyticus ◽  
Sequence Similarity ◽  
Draft Genome ◽  
Draft Genome Sequence ◽  
Toxin Genes ◽  
The One ◽  
Acute Hepatopancreatic Necrosis Disease

We sequenced Vibrio owensii strain SH-14, which causes serious acute hepatopancreatic necrosis disease (AHPND) in shrimp. Sequence analysis showed a large extrachromosomal plasmid, which encoded pir toxin genes and shared highly sequence similarity with the one observed in AHPND-causing Vibrio parahaemolyticus strains. The results suggest that this plasmid appears to play an important role in shrimp AHPND.

scholarly journals Selection of Bacillus spp. isolates having enzyme producing ability and antagonistic to Vibrio parahaemolyticus causing the early mortality syndrome (EMS) on shrimp

2018 ◽  
Vol 1 (6) ◽  
pp. 23-31
Author(s):  
Dung Thi Thanh Do ◽  
Binh Thanh Le ◽  
Duong Thi Dang Hoang ◽  
Quang Dinh Vo ◽  
Trang Thi Phuong Phan
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Extracellular Enzymes ◽  
Early Mortality ◽  
Testing Methods ◽  
Shrimp Ponds ◽  
Early Mortality Syndrome ◽  
Rdna Sequencing ◽  
Potential Applications ◽  
Bacillus Spp ◽  
Selection Of

The aim of this study is to select some Bacillus isolates which are capable of yielding several beneficial enzymes and antagonism to Vibrio parahaemolyticus causing the EMS shrimp disease. In this study, we isolated and screened total of 54 Bacillus isolates from 30 mud, water and shrimp samples at shrimp ponds in Soc Trang province. Among these, 19 isolates were resistant against Vibrio parahaemolyticus strains causing the EMS shrimp disease via two testing methods. Three of them including NA2B13, NA10B2, NA8B1 isolates showed strongresistance and strong one to three kinds of extracellular enzymes to produce. Result of 16S rDNA sequencing and MALDI -TOF showed that NA2B13 and NA8B1 were Bacillus subtilis and NA10B2 was B. amyloliquefaciens. These two species were regarded safe and having potential applications in the production of biological products to prevent EMS shrimp disease.

scholarly journals INVESTIGATING THE PRODUCTION OF EXTRACELLULAR ENZYMES OF VIBRIO PARAHAEMOLYTICUS STRAINS ISOLATED IN VIETNAM

2018 ◽  
Vol 15 (4) ◽  
pp. 703-710
Author(s):  
Ngo Nguyen Vu ◽  
Pham Thi Thu Hien ◽  
Le Nguyen Minh Tien ◽  
Do Ngoc Phuc Chau ◽  
Hoang Tung ◽  
...  
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Extracellular Enzymes ◽  
Extracellular Enzyme ◽  
Negative Bacterium ◽  
Southern Vietnam ◽  
Core Enzyme ◽  
Short Period ◽  
Pcr Method ◽  
Encoding Gene ◽  
Acute Hepatopancreatic Necrosis Disease

Acute Hepatopancreatic Necrosis Disease (AHPND) is a widespread disease targeting cultured shrimps, which results in heavy losses in many regions around the world including Vietnam. The causative agent of the disease is Vibrio paraheamolyticus which is a Gram-negative bacterium present ubiquitously in marine environment. However, this normal floral organism once acquired a plasmid containing Pir- toxin encoding gene (AHPND plasmid) can transform into a highly toxic strain (AHPND strain) with the ability to kill a large number of cultured shrimps in a short period. To understand more on the virulence of V. parahaemolyticus, 17 different V. parahaemolyticus isolates collected from different locations in Southern Vietnam have been analyzed for the presence of AHPND plasmid using PCR method and their extracellular enzyme profile using agar- based method. Seven enzymes which are known to be important virulence factors of the bacterium were studied including caseinase, gelatinase, lecithinase, hemolysin, elastase, lipase and chitinase. Results showed that among 17 studied isolates, six were detected to have AHPND plasmid. Enzymatic activity was observed for caseinase, gelatinase and lecithinase while undetectable for others. The ability to produce these three enzymes varied among different V. parahaemolyticus isolates. Lecithinase appeared in all isolates while the presence of caseinase and gelatinase varied. This indicated that lecithinase seems to be core enzyme of V. paraheamolyticus and the extracellular enzymes do not correlate with the presence of AHPND plasmid. In conclusion, our data suggested that the virulence of Vibrio parahaemolyticus mostly depends on the presence of AHPND plasmid other than extracellular enzymes.

scholarly journals Prevalence and Antimicrobial Resistance of Vibrio spp. in Retail and Farm Shrimps in Ecuador

2015 ◽  
Vol 78 (11) ◽  
pp. 2089-2092 ◽  
Author(s):  
L. SPERLING ◽  
T. ALTER ◽  
S. HUEHN
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Antimicrobial Agents ◽  
Vibrio Vulnificus ◽  
Common Species ◽  
Retail Markets ◽  
Early Mortality Syndrome ◽  
Resistance Patterns ◽  
Acute Hepatopancreatic Necrosis Disease ◽  
Shrimp Farms ◽  
Vibrio Spp

The aim of this study was to investigate the prevalence of Vibrio spp. in shrimp at retail and in shrimp farms in Ecuador and to determine the antimicrobial agent resistance patterns of farm isolates. The presence of genes linked to early mortality syndrome (EMS) or acute hepatopancreatic necrosis disease (AHPND) also was evaluated. Vibrio spp. were isolated from retail shrimps in Cuenca, Ecuador, and farm shrimps originating from provinces El Oro and Guayas, Ecuador. A total of 229 shrimp samples were collected, of which 71 originated from retail markets in Cuenca and 158 came from shrimp farms. Overall, 219 (95.6%) samples tested positive for Vibrio spp. Vibrio parahaemolyticus (80.8%) was the most common species detected, followed by Vibrio alginolyticus (50.2%), Vibrio cholerae (11.3%), and Vibrio vulnificus (3.5%). None of the V. parahaemolyticus isolates carried the virulence-associated tdh and trh genes. In V. parahaemolyticus shrimp farm isolates, high resistance was found to ampicillin (92.2%), and intermediate resistance was found to tetracycline (51.3%) and amikacin (22.1%). Of the V. parahaemolyticus strains, 68 were resistant to at least three antimicrobial agents, and 2 were resistant to seven antimicrobial agents simultaneously. Up to 18 resistant isolates were found for V. alginolyticus, whereas V. vulnificus and V. cholerae isolates were more susceptible. None of the V. parahaemolyticus isolates carried the EMS-AHPND plasmid. The results of this study revealed the ubiquitous occurrence of Vibrio spp. in shrimps at retail and on shrimp farms in Ecuador.

scholarly journals A Natural Vibrio parahaemolyticus ΔpirAVppirBVp+ Mutant Kills Shrimp but Produces neither PirVp Toxins nor Acute Hepatopancreatic Necrosis Disease Lesions

2017 ◽  
Vol 83 (16) ◽  
Author(s):  
Kornsunee Phiwsaiya ◽  
Walaiporn Charoensapsri ◽  
Suwimon Taengphu ◽  
Ha T. Dong ◽  
Pakkakul Sangsuriya ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Vibrio Parahaemolyticus ◽  
Gene Sequence ◽  
Culture Broth ◽  
Upstream Region ◽  
Enzyme Linked Immunosorbent Assay ◽  
Content Type ◽  
Early Mortality Syndrome ◽  
Reverse Transcription Pcr ◽  
Acute Hepatopancreatic Necrosis Disease

ABSTRACT Acute hepatopancreatic necrosis disease (AHPND) of shrimp is caused by Vibrio parahaemolyticus isolates (VPAHPND isolates) that harbor a pVA plasmid encoding toxins PirA Vp and PirB Vp . These are released from VPAHPND isolates that colonize the shrimp stomach and produce pathognomonic AHPND lesions (massive sloughing of hepatopancreatic tubule epithelial cells). PCR results indicated that V. parahaemolyticus isolate XN87 lacked pirA Vp but carried pirB Vp . Unexpectedly, Western blot analysis of proteins from the culture broth of XN87 revealed the absence of both toxins, and the lack of PirB Vp was further confirmed by enzyme-linked immunosorbent assay. However, shrimp immersion challenge with XN87 resulted in 47% mortality without AHPND lesions. Instead, lesions consisted of collapsed hepatopancreatic tubule epithelia. In contrast, control shrimp challenged with typical VPAHPND isolate 5HP gave 90% mortality, accompanied by AHPND lesions. Sequence analysis revealed that the pVA plasmid of XN87 contained a mutated pirA Vp gene interrupted by the out-of-frame insertion of a transposon gene fragment. The upstream region and the beginning of the original pirA Vp gene remained intact, but the insertion caused a 2-base reading frameshift in the remainder of the pirA Vp gene sequence and in the downstream pirB Vp gene sequence. Reverse transcription-PCR and sequencing of 5HP revealed a bicistronic pirAB Vp mRNA transcript that was not produced by XN87, explaining the absence of both toxins in its culture broth. However, the virulence of XN87 revealed that some V. parahaemolyticus isolates carrying mutant pVA plasmids that produce no Pir Vp toxins can cause mortality in shrimp in ponds experiencing an outbreak of early mortality syndrome (EMS) but may not have been previously recognized to be AHPND related because they did not cause pathognomonic AHPND lesions. IMPORTANCE Shrimp acute hepatopancreatic necrosis disease (AHPND) is caused by Vibrio parahaemolyticus isolates (VPAHPND isolates) that harbor the pVA1 plasmid encoding toxins PirA Vp and PirB Vp . The toxins are produced in the shrimp stomach but cause death by massive sloughing of hepatopancreatic tubule epithelial cells (pathognomonic AHPND lesions). V. parahaemolyticus isolate XN87 harbors a mutant pVA plasmid that produces no Pir toxins and does not cause AHPND lesions but still causes ∼50% shrimp mortality. Such isolates may cause a portion of the mortality in ponds experiencing an outbreak of EMS that is not ascribed to VPAHPND. Thus, they pose to shrimp farmers an additional threat that would be missed by current testing for VPAHPND. Moribund shrimp from ponds experiencing an outbreak of EMS that exhibit collapsed hepatopancreatic tubule epithelial cells can serve as indicators for the possible presence of such isolates, which can then be confirmed by additional PCR tests for the presence of a pVA plasmid.

scholarly journals The infection of Vibrio parahaemolyticus in shrimp and human

2018 ◽  
Vol 1 (1) ◽  
pp. 44 ◽  
Author(s):  
Rian Ka Praja
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Animal Health ◽  
Alternative Therapy ◽  
Penaeus Monodon ◽  
Early Mortality ◽  
Economic Losses ◽  
Early Mortality Syndrome ◽  
Thermostable Direct Hemolysin ◽  
Acute Hepatopancreatic Necrosis Disease ◽  
Very High

<p class="15" align="justify"><em>Vibrio parahaemolyticus</em> is an aquatic zoonotic agent that can threaten human and aquaculture animal health. Humans can be infected by consuming contaminated raw seafood or wound-related infections. Generally infection of <em>V. parahemolyticus</em> is orally transmitted and causes gastroenteritis in humans while in aquaculture animals especially shrimp can cause Acute Hepatopancreatic Necrosis Disease (AHPND) or Early Mortality Syndrome (EMS) with a very high mortality rate and cause economic losses. Shrimp species susceptible to infection are <em>Litopenaeus vannamei, Penaeus monodon,</em> and <em>P. chinensis</em>. <em>V. parahaemolyticus</em> produces several toxins in human disease such as thermostable direct hemolysin (TDH), TDH-related haemolysin (TRH), and thermolabile hemolysin (TLH). Meanwhile, Photorabdus insect-related (Pir) toxins consisting of PirA<sup>vp</sup> and PirB<sup>vp</sup> are the toxins associated with AHPND in shrimp. The genes that encode the toxin are used as targets to diagnose <em>V. parahaemolyticus</em> pathogens molecularly. Until now the treatment of <em>V. parahaemolyticus</em> infection is using antibiotics and fluid therapy, but there were <em>V. parahaemolyticus</em> isolates from aquaculture that have been resistant to antibiotics so that the use of antibiotics in aquaculture must be controlled and the use of alternative therapy are very important to be developed to control <em>V. parahaemolyticus</em> infection.</p><p class="15" align="justify"> </p><p>Keywords: <em>V. parahaemolyticus</em>, zoonotic, gastroenteritis, Acute Hepatopancreatic Necrosis Disease (AHPND), Early Mortality Syndrome (EMS).</p>

scholarly journals NGHIÊN CỨU KHẢ NĂNG KHÁNG VI KHUẨN Vibrio spp. GÂY BỆNH HOẠI TỬ GAN TỤY CẤP (AHPND) TRÊN TÔM CHÂN TRẮNG (Litopenaeus vannamei) CỦA DỊCH CHIẾT TỪ CÂY CHÓ ĐẺ THÂN XANH (Phyllanthus amarus Schum)

2019 ◽  
Vol 128 (1E) ◽  
pp. 99-106
Author(s):  
Trần Vinh Phương ◽  
Hoàng Thị Ngọc Hân ◽  
Đặng Thanh Long ◽  
Phạm Thị Hải Yến ◽  
Nguyễn Quang Linh
Keyword(s):  
Litopenaeus Vannamei ◽  
Vibrio Parahaemolyticus ◽  
Early Mortality ◽  
Phyllanthus Amarus ◽  
Early Mortality Syndrome ◽  
Acute Hepatopancreatic Necrosis Disease ◽  
Vibrio Spp

Kết quả nghiên cứu nhằm xác định khả năng kháng khuẩn của dịch chiết từ cây chó đẻ thân xanh (Phyllanthus amarus); nồng độ ức chế tối thiểu (MIC) và nồng độ tiêu diệt tối thiểu (MBC) trên 2 chủng vi khuẩn Vibrio parahaemolitycus (K15) và Vibrio sp. (VT33) ở mật độ 106 CFU/mL gây bệnh hoại tử gan tụy cấp (Acute Hepatopancreatic Necrosis Disease –AHPND) còn được gọi là hội chứng tôm chết sớm (Early Mortality Syndrome - EMS) trên tôm chân trắng (Litopenaeus vannamei) nuôi tại Thừa Thiên Huế, kết quả ban đầu ghi nhận: dịch chiết được chiết xuất dạng cao từ cây chó đẻ thân xanh (P. amarus) có khả năng kháng cả 2 chủng Vibrio parahaemolitycus (K15) và Vibrio sp. (VT33) thử nghiệm từ nồng độ 250 đến 1.000 mg/mL, thể hiện ở đường kính vòng kháng khuẩn dao động từ 16,60–21,40 mm đối với chủng Vibrio parahaemolitycus (K15) và từ 17,60 – 23,60 mm đối với Vibrio sp. (VT33). Kết quả cũng xác định hiệu quả của dịch chiết trên 2 chủng Vibrio parahaemolyticus (K15) và Vibrio sp. (VT123) thể hiện giá trị MIC và MBC tương ứng đạt 125 mg/mL và 500 mg/mL; 62,5 và 500 mg/mL.

scholarly journals Effects of pH, temperature and oxygen-limited condition on the virulence of Vibrio parahaemolyticus

2020 ◽  
Vol 23 (3) ◽  
pp. First
Author(s):  
Van Nhi Tran ◽  
Phuong Nhat Vi Nguyen ◽  
Thi Thu Hoai Nguyen
Keyword(s):  
Enzymatic Activity ◽  
Vibrio Parahaemolyticus ◽  
Extracellular Enzymes ◽  
Brain Heart Infusion ◽  
Effect Of Temperature ◽  
Oxygen Level ◽  
Shrimp Culture ◽  
Gelatinase Activity ◽  
Limited Condition ◽  
Extracellular Enzymatic Activity

Introduction: Vibrio parahaemolyticus is a popular Gram-negative bacterium in the marine and estuarine regions. It can cause Early Mortality Syndrome (EMS), now named Acute Hepatopancreatic Necrosis Disease (AHPND), which resulted in severe losses to the shrimp culture. This study aimed to investigate the effect of pH, temperature, and oxygen-limited condition on the extracellular enzymatic activity of V. parahaemolyticus. Methods: V.  parahaemolyticus XN9, an AHPND-causing strain, was cultured in Brain Heart Infusion (BHI) medium at different pHs (7.5, 8.0, 8.5 and 9.0), temperatures (25oC, 30oC, and 35oC) and different oxygen conditions (either 120rpm shaking or static with the presence of oxygen absorber packages). The activity of five extracellular enzymes, including caseinase, lecithinase, chitinase, gelatinase, and lipase, was assessed using the agar-based method with the corresponding media. Results: When pH was increased from 7.5 to 9.0, caseinase and lipase activity was decreased significantly by 88% and 44%. In contrast, gelatinase activity increased markedly from 0 to 1.38 ± 0.17 (+) mm, and lecithinase reached the highest activity, which was 2.96 ± 0.13 mm (++) at pH 8.5. Regarding effect of temperature, highest activity of caseinase (0.85 ± 0.13 mm (+)) and gelatinase (1.37 ± 0.25 mm (+)) was obtained at 350C, lecithinase at 30oC and lipase at 25oC. Regarding the effect of oxygen level, the activity of most tested enzymes decreased significantly following the decrease of oxygen level. The highest activity of caseinase, gelatinase, and lipase was observed when the bacteria were cultured and tested in a fully oxygenated condition while lecithinase showed the highest activity when the bacteria were cultured in oxygenated condition but tested in oxygen-limited condition. No chitinase activity was observed in any of the tested conditions. Conclusion: Our data suggested that extracellular enzymatic activity of V. parahaemolyticus is significantly influenced by environmental conditions. No particular testing condition resulted in the highest activity for all tested enzymes. However, warm temperature (30/ 35oC), mildly alkaline pH (pH 8.0), and fully oxygenated condition could increase the overall extracellular enzymatic activity of V. parahaemolyticus, thus increase its potential virulence.

Sign in / Sign up

Export Citation Format

Share Document