scholarly journals Detection of Vibrio harveyi using hemolysin primer in tiger shrimp Penaeus monodon

2015 ◽  
Vol 12 (2) ◽  
pp. 101
Author(s):  
Irma Suriyani ◽  
Ince Ayu Khairana Kadriah ◽  
Ilmiah Kuruseng
Keyword(s):  
Rapid Detection ◽  
Vibrio Harveyi ◽  
Penaeus Monodon ◽  
Chain Reaction ◽  
Immersion Method ◽  
Tiger Shrimp ◽  
Hemolysin Gene ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Pcr Techniques

<p class="BasicParagraph" align="center"><strong>ABSTRACT</strong></p><p class="BasicParagraph"> </p><p class="BasicParagraph">This study was aimed to analyze the sensitivity and ability of primer hemolysin in detecting pathogenetic <em>Vibrio</em> on tiger shrimp post-larvae (PL) exposed under different exposure times in media inoculated with <em>Vibrio harveyi</em>. The PL of tiger shrimp were infected with 10<sup>6</sup> cfu/mL of <em>V. harveyi</em> by immersion method for three, six, 12, 24, 48 and 72 hours. The presence of hemolisin genes was detected by PCR techniques. The electrophoresis detected narrow hemolysin genes after PL were exposed for three and six hours. Clear visible bands of DNA <em>Vibrio</em> were observed for 12 hours of exposure. In contrast, no detected hemolysin gene of <em>Vibrio</em> was observed for PL exposed within 24, 48, and 72 hours. The rapid detection on <em>Vibrio</em> pathogenic for tiger shrimp PL should be conducted within three to 12 hours of exposure. No recommendation in utilizing this rapid detection for tiger shrimp PL exposed beyond 12 hours of <em>V. harveyi</em>.</p><p class="BasicParagraph"> </p><p class="BasicParagraph">Keywords: specific primer, luminous <em>Vibrio</em> bacteria, pathogenic, PCR method, hemolysin gene</p><p class="BasicParagraph"> </p><p class="BasicParagraph"> </p><p class="BasicParagraph" align="center"><strong>ABSTRAK</strong></p><p class="BasicParagraph"> </p><p class="BasicParagraph">Penelitian ini bertujuan untuk mengetahui kemampuan atau sensitivitas primer hemolisin dalam mendeteksi <em>Vibrio</em> patogen dengan lama pemaparan berbeda. Penelitian ini dilakukan dengan menginfeksikan <em>Vibrio harveyi</em> pada benur udang dengan metode perendaman pada konsentrasi 10<sup>6</sup> cfu/mL. Pengambilan sampel dilakukan pada waktu tiga, enam, 12, 24, 48, dan 72 jam pascainfeksi. Keberadaan gen hemolisin pada bakteri <em>V. harveyi</em> dideteksi menggunakan teknik <em>polymerase chain reaction </em>(PCR). Hasil elektroforesis memperlihatkan bahwa pada pemaparan tiga dan enam jam keberadaan gen hemolisin dari bakteri <em>Vibrio</em> patogen yang diinfeksikan sudah dapat terdeteksi pada benur walaupun masih terlihat tipis. Pada pemaparan 12 jam terlihat sangat jelas pita-pita DNA dari bakteri patogen. Sedangkan pada pemaparan 24, 48, dan 72 jam sudah tidak terdeteksi lagi gen hemolisin dari bakteri <em>Vibrio</em>. Hal ini diduga disebabkan terjadinya penurunan populasi bakteri <em>Vibrio</em> yang hidup dalam tubuh dan media pemeliharaan udang. Pentingnya deteksi cepat diawal udang terinfeksi bakteri (3–12 jam) karena setelah 12 jam infeksi sudah sulit untuk mendeteksi keberadaan bakteri patogen di dalam tubuh udang.</p><p class="BasicParagraph"> </p><p>Kata kunci: primer spesifik, bakteri <em>Vibrio</em> berpendar, patogenik, metode PCR, gen hemolisin</p><p> </p>

Evaluation of the Polymerase Chain Reaction Method for Identification of Vibrio vulnificus Isolated from Marine Environments

1997 ◽  
Vol 60 (1) ◽  
pp. 81-83 ◽  
Author(s):  
EIJI AONO ◽  
HARUO SUGITA ◽  
JUNJIRO KAWASAKI ◽  
HIROSHI SAKAKIBARA ◽  
TAKUMI TAKAHASHI ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Hybridization ◽  
Vibrio Vulnificus ◽  
Polymerase Chain Reaction Method ◽  
Tokyo Bay ◽  
Phenotypic Characteristics ◽  
Chain Reaction ◽  
Hemolysin Gene ◽  
Pcr Method ◽  
Polymerase Chain

The polymerase chain reaction (PCR) method for identification of Vibrio vulnificus in the marine environment was evaluated by comparing it to both the conventional and DNA-DNA hybridization methods. Of 13,325 isolates obtained from seawater and sediment samples, and oyster and goby specimens collected from the coastal waters of Tokyo Bay, Japan, only 61 isolates were identified as V. vulnificus on the basis of phenotypic characteristics and the amplification of the cytotoxin-hemolysin gene by the PCR method. All 61 isolates were further confirmed to be V.vulnificus by a DNA-DNA hybridization method and the API 20E system although they were divided into 13 groups on the basis of their API 20E profiles. These results strongly suggest that the PCR method is useful for identification of this organism.

KUALITAS INDUK UDANG WINDU (Penaeus monodon) DI PERAIRAN PANTAI PANGANDARAN, SELATAN JAWA BARAT

2012 ◽  
Vol 12 (1) ◽  
Author(s):  
Suhendar I Sachoemar ◽  
Ratu Siti Aliah
Keyword(s):  
Coastal Area ◽  
National Level ◽  
Penaeus Monodon ◽  
Necrosis Virus ◽  
West Java ◽  
Black Tiger Shrimp ◽  
Tiger Shrimp ◽  
Pcr Method ◽  
Polymerase Chain

An evaluation of the quality of the Black Tiger Broodstock Shrimp (Penaeus monodon) in the southern coastal area of Pangandaran, West Java was conducted during period of Juny-October 2005. The evaluation was focused on the waterquality identification and the quality of the broodstock collected from the farmer sorround the Pangandaran Area. The result of the evaluation showed that the water quality of the coastal area of Pangandaran was good compare to the northern coastal area of West Java in which the organic and inorganic pollutantwas . This situation was affected on the quality of the  broodstock. The identification result on the quality of broodstock using polymerase chain reaction (PCR) method shows that the broodstock shrimp of the Black Tiger Shrimp(Penaeus monodon) in the coastal area of Pangandaran is free from the White Spot Syndrome Virus (WSSV) and Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV). From 99 broodstock identified, only 4 indicated by very light contaminated WSSV and 2 by IHHNV. This condition shows that in general the quality of the broodstock shrimp of the Black Tiger Shrimp (Penaeus monodon) in the coastal area of Pangandaran was excellent and these sources are potentialto be applied as broodstock sources for developing the Black Tiger Shrimp (Penaeus monodon) culture in the national level.

scholarly journals Detection of Aedes aegypti Resistance towards Cypermethrin using Polymerase Chain Reaction (PCR) Techniques in Ambarawa Semarang Regency

2020 ◽  
Vol 9 (1) ◽  
pp. 71-77
Author(s):  
Widya Hary Cahyati ◽  
Laila Fitriani
Keyword(s):  
Polymerase Chain Reaction ◽  
Aedes Aegypti ◽  
Molecular Detection ◽  
Sampling Techniques ◽  
Chain Reaction ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Homozygous Resistant ◽  
Development Center ◽  
Pcr Techniques

Ambarawa Regency is one of the endemic sub-districts since 2017 which always accounts for the highest DHF cases in Semarang Regency. Molecular detection of Aedes aegypti resistance towards sipermetrin using Polymerase Chain Reaction (PCR) techniques. The study was conducted to see mutations in the Aedes aegypti VGSC gene. This research was a pure experimental study. The Aedes aegypti samples examined were 10 in every village taken from 3 endemic DHF villages with high fogging intensity in Ambarawa. The research used random sampling techniques taken with ovitrap which was first installed during the month of August. Resistance detection test using the PCR method were conducted at the Banjarnegara Research and Development Center in September. Data were analyzed descriptively to illustrate the results of the study. The results showed that in Tambakboyo Village, 2 samples were susceptible (V / V), 7 samples were detected as homozygous (G / G) resistant, 1 sample was detected as heterozygous (V / G) resistant; in Kupang District 5 samples were detected as being homozygous resistant (G / G) and 5 samples were detected as heterozygous resistant (V / G); and in Panjang Village 1 susceptible sample (V / V), 8 samples were detected as homozygous resistant (G / G), 1 sample was detected as heterozygous resistant (V / G). Based on the results of mutation studies had been found in the VGSC gene in codon V1016G. The proper implementation of management, selection and rotation of insecticides is expected to reduce the risk of resistance in the Aedes aegypti mosquito population.

scholarly journals White Spot Syndrome Virus (WSSV) and Vibrio sp. in the Fresh Feed Used as Tiger Shrimp, Penaeus monodon, Broodstock Diet

2007 ◽  
Vol 3 (1) ◽  
pp. 19
Author(s):  
R.W. Haliman
Keyword(s):  
Polymerase Chain Reaction ◽  
White Spot Syndrome Virus ◽  
Penaeus Monodon ◽  
White Spot ◽  
Chain Reaction ◽  
Infection Level ◽  
Tiger Shrimp ◽  
Polymerase Chain ◽  
White Spot Syndrome ◽  
Broodstock Diet

<p>The experiment was conducted to evaluate the presence of white spot syndrome virus (WSSV) and <em>Vibrio </em>sp. in clam, oyster crabs, squid and sea worm as fresh feed for tiger shrimp, <em>Penaeus monodon, </em>Broodstock. The presence of WSSV was detected by using polymerase chain reaction (PCR) method, while the <em>Vibrio </em>sp. was grown in TCBS agar and counted as cfu/g fresh feed. The result shows that all the feed have already infected by WSSV with light infection level. <em>Vibrio </em>sp. can be isolated from all samples and their population were 1.5 x 103 - 1,8 x 104 cfu/g fresh feed.</p> <p>Key words: WSSV, <em>Vibrio </em>sp., fresh feed, tiger shrimp broodstock</p> <p> </p> <p>ABSTRAK</p> <p>Penelitian ini dilakukan untuk mengetahui keberadaan <em>white spot syndrome virus </em>(WSSV) dan <em>Vibrio </em>sp. pada pakan segar untuk induk udang windu. <em>Penaeus monodon. </em>Pakan segar yang dievaluasi terdiri dari kerang, tiram, kepiting, cumi-cumi dan cacing laut. WSSV dideteksi dengan metode <em>polymerase chain reaction </em>(PCR). Kandungan <em>Vibrio </em>sp. dihitung dengan menginokulasikan 0,1 ml suspensi pakan segar pada agar TCBS. kemudian diinkubasikan pada suhu ruang selama 24 jam. Koloni bakteri yang tumbuh dihitung dan dinyatakan dalam cfu/g pakan. Hasil pengamatan menunjukkan bahwa semua jenis pakan segar terinfeksi WSSV pada skala ringan. <em>Vibrio </em>sp. dapat diisolasi dari semua jenis pakan segar, dan jumlah populasinya 1,5 x 103- 1,8 x 104 cfu/g pakan.</p> Kata kunci: WSSV, <em>Vibrio </em>sp., pakan segar, induk udang windu

scholarly journals A comparative evaluation of two chromogenic media for surveillance of carbapenem-resistant Enterobacterales with non-carbapenemase-producing or carbapenemase-producing strains other than blaKPC: blaGES-5, blaNDM-1 or blaVIM-2

2019 ◽  
Vol 43 (3) ◽  
pp. 173-176
Author(s):  
Chang-Hun Park
Keyword(s):  
Rapid Detection ◽  
Comparative Evaluation ◽  
Susceptibility Testing ◽  
Rapid Identification ◽  
Surveillance Program ◽  
Contact Precaution ◽  
Chain Reaction ◽  
Carbapenem Resistant ◽  
Control And Prevention ◽  
Polymerase Chain

Abstract Background Infections caused by carbapenem-resistant Enterobacterales (CREs) are an emerging problem associated with high rates of morbidity and mortality. CREs are divided into two categories (carbapenemase-producing [CP] CRE and non-CP CRE). The most prevalent carbapenemase produced by Enterobacterales is Klebsiella pneumoniae carbapenemase (KPC) in Korea. Rapid identification of CREs is clinically important in infection control precaution. We compared the performance of two chromogenic media (chromID CARBA agar and CHROMagar KPC agar) for non-CP CREs or CP CREs with blaGES-5, blaNDM-1 or blaVIM-2 in a Korean hospital. Methods The study was carried out during a 3-month period from April to June 2017 during the surveillance program for CRE colonization. Antimicrobial susceptibility testing (AST) and polymerase chain reaction (PCR) were performed at the Korean Centers for Disease Control and Prevention. Results A total of 45 rectal swabs from 42 hospitalized patients were examined. Sensitivity of both chromID CARBA and CHROMagar KPC were 100% for CP CREs; and 50% and 100% for non-CP CREs, respectively. Specificity of chromID CARBA and CHROMagar KPC were 89.2% and 70.3% for CP CRE, respectively; and 76.9% and 66.7% for non-CP CRE, respectively. Conclusions The CHROMagar KPC is useful to monitor non-CP and CP CREs. The chromID CARBA is efficient for rapid detection of CP CREs requiring high contact precaution.

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