• Journal of Microbiology and Biotechnology
• /
• 제22권10호
• /
• pp.1350-1358
• /
• 2012

Viral pathogens, alongside other pathogens, have major effects on crustacean aquaculture. Hepatopancreatic parvovirus (HPV) is an emerging virus in the shrimp industry and has been detected in shrimp farms worldwide. The HPV genome has greater diversity than other shrimp viruses owing to its wide host range and geographical distribution. Therefore, developing diagnostic tools is essential to detect even small copy numbers from the target region of native HPV isolates. We have developed two easy to use quantitative real-time PCR kits, called Green Star and Dual Star, which contain all of the necessary components for real-time PCR, including HPV primers, using the primers obtained from the sequences of HPV isolates from Korea, and analyzed their specificity, efficiency, and reproducibility. These two kits could detect from 1 to $1{\times}10^9$ copies of cloned HPV DNA. The minimum detection limits obtained from HPV-infected shrimp were $7.74{\times}10^1$ and $9.06{\times}10^1$ copies in the Green Star and Dual Star assay kits, respectively. These kits can be used for rapid, sensitive, and efficient screening for HPV isolates from Korea before the introduction of postlarval stages into culture ponds, thereby decreasing the incidence of early development of the disease.

• 한국어병학회지
• /
• 제25권3호
• /
• pp.271-277
• /
• 2012

2012년 5월에서 11월 사이에 우리나라 넙치, 조피볼락, 무지개송어, 뱀장어, 흰다리새우, 전복 양식장에서 발생하는 폐사피해를 표본조사를 통해 모니터링하였다. 조사의 방식은 해당 양식장과 거래하고 있거나 관계를 맺고 있는 전국의 수산질병관리사 조사요원의 직접 현장 방문을 통한 면접과 표준 매뉴얼을 통한 진단을 통하여 실시하였다. 조사기간 중 넙치 양식장에서는 총 27.9%의 누적 피해율을 보여주었고, 이중 스쿠티카병, 연쇄구균병, VHS, 비감염성 원인에 의한 폐사, 비브리오병, 활주세균병의 순으로 폐사피해가 높은 것으로 나타났다. 조피볼락 양식장에서는 13.6%의 누적 피해율을 보여주었고, 아가미 흡충병, 연쇄구균병, 비브리오병, 활주세균병, 비감염성 원인에 의한 폐사 순의 폐사피해가 나타났다. 무지개송어의 경우 총 17.1%의 피해율을 기록하였고, 비감염성 원인에 의한 폐사와 백점병이 주된 폐사 원인으로 집계되었다. 뱀장어의 경우 비교적 낮은 0.6%의 피해율을 기록하였으며, 에드워드병, 백점병, 아가미흡충병이 주된 피해요인이었다. 흰다리새우는 71.2%의 피해율을 보였으며, 대부분 비감염성 원인에 의한 폐사으로 집계되었고, 바이러스성 흰반점병도 12.5%의 피해를 준 것으로 나타났다. 전복의 경우 10.7%의 피해율 모두가 비감염성 원인에 의한 폐사 피해인 것으로 조사되었다.

• 한국수산과학회지
• /
• 제52권6호
• /
• pp.656-665
• /
• 2019

Environmental monitoring was conducted to identify the characteristics and patterns of sedimentary environments of tidal flats in 12 areas on the west coast of Korea. The habitat of the mud shrimp Upogebia major contained higher organic and mud contents compared to the habitat of the manila clam Ruditapes philippinarum. In particular, the mud content increased from 84.38% to 89.18% in the Seongam-4 area, where the mud shrimp and manila clam coexist, and the mean grain size exhibited a finer particle size, from 5.48 Φ to 5.80 Φ. In the mud shrimp habitat, the sediment mud content was > 60% and the mean grain size was > 5 Φ. Additionally, the mud shrimp only inhabited open coast tidal flat areas. The management of shellfish aquaculture farms by physical methods should be continued based on comparison of the sedimentary environments in the Boryeong and Seongam areas in response to the damage to the mud shrimp habitat.

• 한국어병학회지
• /
• 제28권1호
• /
• pp.1-7
• /
• 2015

In 2009 outbreaks of an emerging disease in shrimp farms were first reported in China. The disease was known as early mortality syndrome (EMS), or acute hepatopancreatic necrosis disease (AHPND). Since 2009, the disease has been spread to Vietnam, Malaysia, Thailand and Mexico. The causative agent of the disease was identified as Vibrio parahaemolyticus. It is a common seawater inhabitant bacterium, and the pathogen can sometimes contaminate seafood. The disease has caused mass mortality of cultivated shrimp, and huge economic losses in the countries named above. In order to prevent the introduction and establishment of AHPND, emergency measures, such as strengthening of import conditions and even import bans, were put in place by many other countries. In Korea, on the other hand, there are large quantities of shrimp imports from countries, such as Thailand and Vietnam. Transportation of live and fresh dead shrimp is highly likely, and could be a transmission pathway if the shrimp are sourced from populations in AHPND endemic areas. It is important to recognize that importing countries may provisionally adopt sanitary or phytosanitary measures on the basis of available pertinent information, including that from the relevant international organizations, as well as from sanitary or phytosanitary measures applied by other countries based on "Agreement on Application of Sanitary and Phytosanitary Measures". It is pertinent that Korea also takes proper emergent measures to keep out diseases and provide safe seafood.

• 한국어병학회지
• /
• 제27권2호
• /
• pp.91-97
• /
• 2014

수산생물질병관리법이 제정 시행 (2009)된 이후 매년 우리나라 양식산 흰다리새우에 대한 국내 미발생 법정전염병을 모니터링하고 있다. 최근 외래질병의 유입가능성이 증가함에 따라 질병 감시를 강화하고자 조사대상 법정전염병을 2종 (YHD, IMN)에서 5종 (YHD, IMN, IHHN, TS, WTD)으로 확대하고 조사지역과 양식장 개소를 매년 상향 조절하여 실시하였다. 2013년에는 지정 법정전염병 5종에 대해 흰다리새우 종묘생산장 및 양식장 29개소에서 총 2,650마리의 샘플을 조사한 결과, 검사한 모든 시료에서 바이러스가 검출되지 않았다. 새우 양식장에서의 향후 새로운 질병 유입에 대한 위험성을 낮추고 국가적 질병관리 및 차단 방역을 위해서는 지속적인 질병 예찰 및 질병 감시가 필수적일 것이다.

• Fisheries and Aquatic Sciences
• /
• 제11권4호
• /
• pp.195-204
• /
• 2008

Viral diseases are major emerging problems of shrimp that have affected the production, and even complete losses for shrimp farms. In this study, we developed a sensitive TaqMan real-time PCR method to quantify white spot syndrome virus (WSSV) and hepatopancreatic parvovirus (HPV) in the shrimp and pond water in which fleshy shrimp, Fenneropenaeus chinensis, and Pacific white shrimp, Litopenaeus vannamei, are reared. WSSV and HPV in pond seawaters ranged from $1.65{\times}10^3$ to $2.43{\times}10^9$ and from 0 to $4.43{\times}10^5$ copies/L of seawater, respectively. Of 20 ponds analyzed, all pond water and shrimp were positive for WSSv. L. vannamei showed higher susceptibility to WSSV than F chinensis. HPV was detected only in the pond water for F chinensis. In shrimp tissue, however, HPV was found in both species, with 23-times higher infection rate in F chinensis than L. vannamei. The total bacterial counts in the pond water ranged from $2.23{\times}l0^3$ to $1.98{\times}l0^5\;CFU/mL$. The variations in total bacterial count for each pond appeared to correlate to the variations of the WSSV load. Statistical analysis indicated that there was no significant difference (P>0.05) between the WSSV load in pond water and shrimp, and there was no relationship between total bacterial load and viral load in the pond water. However, a significant difference (P<0.01) was found between HPV load and L. vannamei and F chinensis pond water.

• 한국양식학회지
• /
• 제15권1호
• /
• pp.7-13
• /
• 2002

Since 1993, the White Spot Syndrome Virus (WSSV) disease occurred in China among cultured shrimps resulting in mass mortality. Epizootiological surveys undertaken during the outbreak period of 1993-1994 indicated that all stages of Penaeus chinensis, P. japonicus and P. monodon were infected. Consequent to the transport of contaminated shrimp seedlings and seawater, the disease spread all over the farms of China. The disease was more rapidly transmitted at temperatures above $25^{\circ}C$. Challenge experiments showed the causative agent was highly virulent. White spots appeared on the carapace of both span-taneous and experimentally infected shrimps. Moribund shrimps contained turbid hemolymph, hypertrophied Iymphoid organ and a necrotic mid-gut gland. Electron microscopy showed the presence of viral particles in the gills, stomach, lymphoid organ, and epidermal tissue of the infected shrimp. The visions were slightly ovoid with an envelope and averaged 350 $\times$ 150 nm; nucleocapsids measured 375 $\times$ 157 nm. With discontinuous sucrose gradient of 35, 50 and 60% (w/v), the virus was separated from hemolymph of the infected shrimp. The estimated molecular weight of genomic DNA was 237 Kb with EcoR I, 247 Kb with Hind III and 241kb with Pst I. A total of 9 hybridoma colones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with WSSV. The immunofluorescence assay of gill tissue showed that the MAbs reacted with diseased but not with healthy shrimp. The MAbs belonged to IgGl, IgG2b subclass and IgM class, all with kappa light Immune-electron-microscopy with colloidal gold marker showed the presence of 5 MAbs epitopes on the envelope and one on the capsid of the virus. Baculoviral mid-gut gland necrosis showed the specificity of the MAbs produced. For diagnosis 5 different methods were selected. Using Kimura primers for PCR, or MAbs for immunoblot, ELISA or FAT method, in situ hybridization was carried out to show the gene. All these methods detected WSSV in the organ samples of the diseased shrimp but not in healthy one.

• Journal of Microbiology and Biotechnology
• /
• 제11권3호
• /
• pp.394-398
• /
• 2001

Because of the great concern over the possibility of contamination from the rod-shaped nuclear virus (PRDV) from Japan and white spot virus (WSSV) from Taiwan, most eggs used in Korean shrimp farms are currently obtained from local broodstock. In addition, the screening of imported broodstock for any viral presence at the National Fisheries Research and Development Institute is also mandatory. Nonetheless, massive mortality from white spot syndrome continues in Korea. In the present study, we present an improved PCR method to use tissue-extracted DNA instead of viral DNA extracted from a purified virus based on a sucrose density gradient, and produced results within 8 h. In 1998, this modified PCR method was able to detect that diseased Penaeus japonicus were infected within 8 h. In 1998, this modified PCR method was able to detect that diseased Penaeus japonicus were infected only with PRDV, while Fenneropenaeus chinensis were infected with both PRDV and WSSV. In 1999, PRDV and WSSV were detected in F. chinensis with signs of infection, but not with WSSV alone.

• 한국어병학회지
• /
• 제37권1호
• /
• pp.139-145
• /
• 2024

In this study, nylon mash and silicone tube mainly used as shrimp farm equipment were contaminated with Enterocytozoon hepatopenaei (EHP) which is the cause of Hepatopancreatic microsporidiosis (HPM), and were treated with calcium hypochlorite or UV-C disinfection methods for EHP eradication. As a result, similar with the control group (not disinfected), EHP was detected on the nested PCR until the 10 days in the UV-C single treated group. On the other hand, EHP was not detected from 7 days in calcium hypochlorite single treated group (total concentration 200 ppm as available chlorine), and combination of calcium hypochlorite and UV-C treated group revealed no detection of EHP from 3 days. It is appropriate that treated with UV-C and calcium hypochlorite for 3 days or single treated with calcium hypochlorite for 7 days to eradicate EHP on contaminated instrument used in shrimp farms. In contrast, disinfection effect of only using UV-C is very low.

• 한국양식학회지
• /
• 제20권4호
• /
• pp.270-277
• /
• 2007

흰반점바이러스(WSSV, white spot syndrome virus)는 세계적으로 새우양식산업에 가장 심각한 피해를 주는 질병으로 우리나라에는 1993년 서해안에서 처음 보고된 이래 해마다 축제식 새우양식장에 반복적으로 대량폐사를 가져오고 있다. WSSV는 전염성이 매우 강한 바이러스로서 새우양식장에 내에서 감염된 새우들 간에 공식에 의하여 빠르게 감염이 확산될 수 있다. 황복과 같은 육식성 어류는 감염새우 혹은 약한 새우를 선택적으로 포식함으로써 바이러스의 발병을 지연 혹은 억제시킬 수 있는 것으로 알려져 있어 양식현장에서 새우와 함께 복합적으로 양식이 시도되고 있지만 황복의 감염새우에 대한 선택적 포식의 정도와 효과에 대해서는 보고되어 있지 않다. 본 연구에서는 WSSV에 감염된 새우와 비감염된 건강한 새우를 대상으로 사육생물의 연령과 크기에 따른 황복의 선택적 포식효과를 조사하기 위하여 흰다리새우 Litopenaeus vannamei의 치하와 5개월산 황복, 중형 새우와 16개월산 황복으로 구분하여 원형수조($28.26\;m^2$)에서 사육실험을 실시한 결과, 치하와 5개월산 황복의 복합사육의 경우 건강새우와 감염새우의 생존율은 각각 89.1%, 46.0%였으며 중형새우와 16개월산 황복의 실험에서는 건강새우와 감염새우의 생존율이 각각 48%, 4%로서 모두 감염새우가 건강새우에 비해 생존율이 크게 떨어져 황복은 건강새우보다는 감염새우를 선택적으로 포식하는 것으로 나타났다. 황복은 크기에 관계없이 단위체중당 하루에 포식하는 새우의 총 중량은 크게 차이가 나지 않으며 새우의 크기와 밀도의 차이에 따라서도 차이를 보이지 않았다. 또한 황복은 감염새우를 선택적으로 포식하기는 하지만 감염새우가 부족할 경우에는 새우의 건강상태와는 무관하게 일정량의 새우를 포식하기 때문에 새우양식장에 황복을 넣을 때는 황복의 입식시기와 밀도 뿐 아니라 황복의 총중량도 충분히 고려되어야 한다.