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3 results on '"Piyanuch PROMPAMORN"'

1. Effects of Water Temperature on the White Spot Syndrome Virus Infection in Postlarvae Litopenaeus vannamei

Author

Sutee WONGMANEEPRATEEP, Puttharat BAOPRASERTKUL, Piyanuch PROMPAMORN, Kanittada THONGKAO, Chalor LIMSUWAN, and Niti CHUCHIRD

Subjects
White spot syndrome virus, temperature, infection, Litopenaeus vannamei, postlarvae, Technology (General), T1-995, Science (General), Q1-390
Abstract

This study evaluated the effects of high water temperature (32 ± 1 °C) on the white spot syndrome virus (WSSV) infection in Litopenaeus vannamei postlarvae (PL15). WSSV challenge was done by immersion. One group of PL15 was continuously maintained at 32 ± 1 °C until the end of the experiment after challenge and a control group of PL15 was constantly maintained at 28 ± 1 °C until the end of the experiment after challenge. Other groups were kept at 32 ± 1 °C until temperature was altered from 32 ± 1 °C to 28 ± 1 °C at 0, 1, 3, 5 and 7 days after infection. Gross signs and mortality were monitored every 12 h until the end of the experiment. WSSV infections were confirmed by nested-PCR, histopathology, immunohistochemistry and bioassay methods. Challenged shrimp were kept at 32 ± 1 °C for 0, 1, 3 and 5 days before the temperature was reduced to 28 ± 1 °C revealing that maintaining the temperature at 32 ± 1 °C for a longer period could delay clinical signs and onset of mortalities. Nevertheless, 100 % mortalities occurred in all groups and the control group within 7 days. All moribund PL15 were WSSV-positive by nested-PCR assay as well as histopathology, immunohistochemistry and bioassay methods. In contrast, PL15 constantly maintained at 32 ± 1 °C until the end of the experiment, and for 7 days after challenge before switching to 28 ± 1 °C did not show clinical signs and mortality. Surviving PL15 from both groups were WSSV-negative by nested-PCR assay as well as histopathology, immunohistochemistry and bioassay methods. This study clearly indicated that postlarvae maintained constantly at 32 ± 1 °C for 7 days were able to eliminate/clear WSSV infection.

Published
2011
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2. The development of loop-mediated isothermal amplification combined with lateral flow dipstick for detection of Vibrio parahaemolyticus

Author

Siwaporn Longyant, Parin Chaivisuthangkura, Sombat Rukpratanporn, Paisarn Sithigorngul, Piyanuch Prompamorn, and Pattarin Sridulyakul

Subjects
Detection limit, Vibrio parahaemolyticus, Loop-mediated isothermal amplification, food and beverages, Dipstick, Biology, Amplicon, biology.organism_classification, Applied Microbiology and Biotechnology, Molecular biology, Vibrio, law.invention, law, bacteria, Thermolabile, Polymerase chain reaction
Abstract

Aims: The current study was aimed to develop a loop-mediated isothermal amplification (LAMP) combined with amplicon detection by chromatographic lateral flow dipstick (LFD) assay for rapid and specific detection of Vibrio parahaemolyticus. Methods and Results: Biotinylated LAMP amplicons were produced by a set of four designed primers that recognized specifically the V. parahaemolyticus thermolabile haemolysin (tlh) gene followed by hybridization with an FITC-labelled probe and LFD detection. The optimized time and temperature conditions for the LAMP assay were 90 min at 65°C. The LAMP–LFD method accurately identified 28 isolates of V. parahaemolyticus but did not detect 24 non-parahaemolyticus Vibrio isolates and 35 non-Vibrio bacterial isolates. The sensitivity of LAMP–LFD for V. parahaemolyticus detection in pure cultures was 120 CFU ml−1. In the case of spiked shrimp samples without enrichment, the detection limit for V. parahaemolyticus was 1·8 × 103 CFU g−1 or equivalent to 3 CFU per reaction while that of conventional PCR was 30 CFU per reaction. Conclusions: The established LAMP–LFD assay targeting tlh gene was specific, rapid and sensitive for identification of V. parahaemolyticus. Significance and Impact of the Study: The developed LAMP–LFD assay provided a valuable tool for detection of V. parahaemolyticus and can be used effectively for identification of V. parahaemolyticus in contaminated food sample.

Published
2011
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3. Rapid identification and differentiation of Vibrio parahaemolyticus from Vibrio spp. in seafood samples using developed monoclonal antibodies

Author

Piyanuch Prompamorn, Chalinan Pengsuk, Siwaporn Longyant, Paisarn Sithigorngul, and Parin Chaivisuthangkura

Subjects
Physiology, medicine.drug_class, Cross Reactions, Monoclonal antibody, medicine.disease_cause, Applied Microbiology and Biotechnology, Dot blot immunoassay, Cross-reactivity, Sensitivity and Specificity, Microbiology, Western blot, medicine, Immunoassay, Bacteriological Techniques, biology, medicine.diagnostic_test, Vibrio parahaemolyticus, food and beverages, Antibodies, Monoclonal, General Medicine, biology.organism_classification, Virology, Antibodies, Bacterial, Vibrio, Shrimp, Rapid identification, Seafood, bacteria, Biotechnology
Abstract

Monoclonal antibodies (MAbs) specific to Vibrio parahaemolyticus were successfully generated. According to the specificity of V. parahaemolyticus, MAbs can be classified into 5 groups. The MAbs VP-2D and VP-11H were specific to the O2 and O4 groups of V. parahaemolyticus, respectively. The MAb VP-11B reacted with 11 out of 30 isolates of V. parahaemolyticus used in this study. The MAb VP-516 bound to 27 out of 30 isolates of V. parahaemolyticus and cross reacted with all 10 isolates of V. alginolyticus. The MAb VP-618 demonstrated positive reactivity to 29 out of 30 isolates of V. parahaemolyticus and demonstrated slight cross reactivity to 3 out of 30 isolates of V. harveyi. The sensitivity of the MAbs ranged from 10(8) to 10(7) c.f.u. ml(-1) for V. parahaemolyticus obtained from pure cultures and depended on the group of MAbs. However, the detection capability could be improved to be equivalent to that of the PCR technique following pre-incubation of the samples in alkaline peptone water (APW). Using these MAbs along with MAbs specific to V. alginolyticus (VA-165), V. cholerae (VC-63), V. harveyi (VH-9B and VH-20C) and Vibrio spp. (VC-201) from previous studies, V. parahaemolyticus could be identified and differentiated from Vibrio spp. in various seafood samples including shrimp, green mussels, blood clams and oysters by a simple dot blot immunoassay without the requirement for bacterial isolation or biochemical characterization.

Published
2012

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