AP4 method for two-tube nested PCR detection of AHPND isolates of Vibrio parahaemolyticus

Dangtip, Sirintip; Sirikharin, Ratchanok; Sanguanrut, Piyachat; Thitamadee, Siripong; Sritunyalucksana, Kallaya; Taengchaiyaphum, Suparat; Mavichak, Rapeepat; Proespraiwong, Porranee; Flegel, Timothy W.

doi:10.1016/j.aqrep.2015.10.002

Cited by 136 publications

(115 citation statements)

References 12 publications

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“…sistemas de diagnósticos moleculares basados en el PCR, siendo capaces de detectar la presencia de la cepa portadora de los genes que codifican para las toxinas (OIE, 2013;Nunan et al, 2014;Dangtip et al, 2015;Hong et al, 2015;Sritunyalucksana et al, 2015).…”

Section: Diagnóstico Del Ahpndunclassified

Necrosis aguda del hepatopáncreas: una revisión de la enfermedad en Penaeus vannamei

Mejías

Peña²,

Aranguren

2017

Mesoam. Agron.

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Section: Diagnóstico Del Ahpndunclassified

Necrosis aguda del hepatopáncreas: una revisión de la enfermedad en Penaeus vannamei

Mejías

Peña²,

Aranguren

2017

Mesoam. Agron.

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“…A list of primers to identify AHPND positive V. parahaemolyticus is given in Table 1. Dangtip et al (2015) *Primer AP4-F1 is identical to primer AP3-F from the AP3 method.…”

Section: Methodsmentioning

confidence: 99%

“…ldh (lecithin dependent hemolysin) as described by Taniguchi et al (1985). AP3 and AP4 primers were also used to check the species specific pathogenicity as described by Sirikharin et al (2014) and Dangtip et al (2015), respectively; these primers target contigs that belong to a plasmid in pathogen. A list of primers to identify AHPND positive V. parahaemolyticus is given in Table 1.…”

Section: Methodsmentioning

confidence: 99%

“…The histopathology of AHPND is characterized by massive cell sloughing of HP tubule epithelial cells together with the dysfunction of B, F, R and E cells of HP of affected shrimps (Flegel 2012;Joshi et al 2014). Detection of V. parahaemolyticus isolates is typically based on molecular biological analysis that amplify species-specific gene ldh (lecithin dependent hemolysin) (Taniguchi et al 1985); and to identify the AHPND positive V. parahaemolyticus strains AP3 (Sirikharin et al 2014) and AP4 (Dangtip et al 2015) primers are commonly used.…”

Section: Introductionmentioning

confidence: 99%

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Molecular Identification of AHPND Positive Vibrio Parahaemolyticus causing an outbreak in South-West Shrimp Farming Regions of Bangladesh

Eshik

Abedin

Punom

et al. 2018

J Bangladesh Acad Sci

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Acute hepatopancreatic necrosis disease (AHPND) of shrimps is a bacterial disease, first appeared in China in 2009 and causes mortality up to 100 % which usually occurs early (within approximately first 35 days) after stocking shrimp fry of black tiger shrimp, Penaeus monodon (Fabricius) and white leg shrimp, Litopenaeus vannamei (Boone). The purpose of this study was to isolate and identify the pathogenic strain of V. parahaemolyticus causing AHPND in cultured shrimps (P. monodon) using molecular techniques. After a disease outbreak in April 2017, shrimp samples were collected from three different locations of south-west shrimp farming region of Bangladesh viz. Satkhira, Khulna and Bagerhat districts. In this study, three selective media were used for primary isolation of V. parahaemolyticus. Among 28 primary isolates, representative14isolates were checked for the species-specific detection of V. parahaemolyticus using ldh primers and all of them were found to be positive. The isolates were further characterized to check their AHPND positivity using AP3 and AP4 primers. Four isolates showed positive results for both AP3 and AP4 which indicated that the isolates were AHPND positive. This study also report that all AHPND positive strains showed sensitivity to the antibiotics Tetracycline and Nalidixic Acid. The results of this study will help the farmers and policy makers to make plan to protect shrimps from AHPND and thereby sustain the shrimp farming in Bangladesh.

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“…Mortalities were monitored for 2 d. The moribund and surviving shrimp infected with the strain were collected for histopathological examination and polymerase chain reaction (PCR) of the stomach and HP. Pure isolates of the pathogenic strain (ID 01082013-LS) were molecularly identified by PCR in accordance with the methodology previously described by Flegel & Lo (2014), and the more sensitive AP4 method was also run (Dangtip et al 2015) to detect AHPND. In addition, the typical histopathological lesions of the disease as described by Tran et al (2013) were analyzed.…”

Section: Confirmation Of Bacterial Virulence (Preliminary Bioassay)mentioning

confidence: 99%

Evaluation of medicinal plants and colloidal silver efficiency against Vibrio parahaemolyticus infection in Litopenaeus vannamei cultured at low salinity

Morales-Covarrubias

García-Aguilar²,

Bolan-Mejía³

et al. 2016

Dis. Aquat. Org.

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In shrimp aquaculture, reduction in the use of synthetic antibiotics is a priority due to the high incidence of resistant bacteria (Vibrio) in the white shrimp Litopenaeus vannamei. An increasing number of studies show bactericidal activity of natural treatments in aquaculture. The effectiveness of neem (Azadirachta indica) and oregano (Lippia berlandieri) aqueous extracts and colloidal silver against V. parahaemolyticus were evaluated in low salinity shrimp culture. Results show that aqueous extracts of oregano and neem each present a minimum inhibitory concentration (MIC) of 62.50 mg ml −1 and inhibitory halos of 12.0 to 19.0 mm. Colloidal silver gave a MIC of 2 mg ml −1 , and the inhibitory halos were found to be between 11.8 and 18.8 mm, depending on treatment concentrations. An in vivo challenge test was conducted on white shrimp postlarvae cultured at low salinity (5 practical salinity units, PSU), and a significant increase (p < 0.05) in survival was demonstrated in the presence of the aqueous extracts (oregano 64%, neem 76% and colloidal silver 90%), when compared to the control (0%) in the challenge test. However, no significant differences were ob served between treatments, suggesting that they all act as alternative bactericidal source agents against V. parahaemolyticus infections for L. vannamei postlarvae when cultured at 5 PSU.KEY WORDS: Neem · Oregano · Colloidal silver · Vibrio parahaemolyticus · White shrimp · EMS · Acute hepatopancreatic necrosis syndrome · AHPNS Resale or republication not permitted without written consent of the publisherDis Aquat Org 122: [57][58][59][60][61][62][63][64][65] 2016 organisms for human consumption (Mioso et al. 2014).For over 2000 yr, medicinal plants have been used to combat pathogenic agents, and within the last decades, studies have focused on natural alternatives that can be employed as bactericidal treatment in aquaculture (Torres et al. 2007, Ramírez & Marín-Castaño 2009, García-Aguilar 2013.To transform a medicinal plant for conventional medical use, the most common procedures are infusion and decoction to extract the active substances from the vegetable tissues. When the water ebullition point (100°C) is reached and the water makes contact with fresh or dry plants, the vegetable cells explode and discharge active substances. The scientific term for this is 'aqueous extracts', and the principal advantage of using this preparation method is provision of easy and fast in situ preparations (Citarasu 2010).One plant considered an alternative medicine in aquaculture is neem Azadirachta indica; its principal active metabolite is nortriterpenoide (also known as azadiractine) with the presence of limonoides, which are structurally related and are biologically active (Hammer et al. 1999, López-Pantoja et al. 2007, Banerjee et al. 2013, Dhama et al. 2013a,b, Peña-Navarro et al. 2013. Another plant for consideration is oregano Lippia berlandieri Schauer, which possesses thymol and carvacrol as its bio active substances, which both show significant b...

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AP4 method for two-tube nested PCR detection of AHPND isolates of Vibrio parahaemolyticus

Cited by 136 publications

References 12 publications

Necrosis aguda del hepatopáncreas: una revisión de la enfermedad en Penaeus vannamei

Necrosis aguda del hepatopáncreas: una revisión de la enfermedad en Penaeus vannamei

Molecular Identification of AHPND Positive Vibrio Parahaemolyticus causing an outbreak in South-West Shrimp Farming Regions of Bangladesh

Evaluation of medicinal plants and colloidal silver efficiency against Vibrio parahaemolyticus infection in Litopenaeus vannamei cultured at low salinity

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