Development of monoclonal antibodies against VP28 of WSSV and its application to detect WSSV using immunocomb

White spot disease (WSD) is an important viral disease of penaeid shrimp caused by white spot syndrome virus (WSSV). WSSV isolated from WSD outbreaks in commercial shrimp (Penaeus monodon) farms in India were propagated in the laboratory in healthy shrimp. The virus was purified from the infected tissues by sucrose gradient centrifugation. The VP28 was electroeluted from SDS-PAGE gels and was used to immunize Balb/c mice to produce hybridomas secreting monoclonal antibodies (MAb) against WSSV. A total of five hybridoma clones secreting MAbs to VP28 were produced. The MAbs were of the isotypes IgG1, IgG2b and IgM. The MAbs reacted with VP28 of WSSV and not with any other viral or shrimp protein in western blot. The MAbs were used to develop dot immunoblot assay using an immunocomb to detect WSSV from field samples. The test developed had an analytical sensitivity of 625 pg and a diagnostic sensitivity of 100% compared to single step polymerase chain reaction (PCR). The test can be used as an alternate for first step PCR to detect WSSV from field samples.     

Epidemiological Investigation of Brackish Water Culture Systems in West Bengal,India

An epidemiological investigation was done in brackish water culture systems in three coastal districts of West Bengal. A total of 198 farms were randomly surveyed with a structured questionnaire. The data showed that there was a significant difference in outbreak of white spot disease (WSD) (p < 0.01), shell-associated problems (p < 0.01), and gill-associated problems (p < 0.05) among the culture systems. Among all systems, stunted and uneven growth and white fecal disease (only in shrimp monoculture) were the dominant emerging disorders. WSD remained the most prevalent disease. Some farms tested (polymerase chain reaction [PCR]) positive for WSD, but the animals were apparently healthy. Chlorination, use of PCR screening, application of immunostimulants, and strict bio-security measures play major roles in containing disease outbreaks.     

Breeding for disease resistance of Penaeid shrimps

Diseases are a major constraint on the intensive production of shrimps. Conditions in production ponds favour disease development, and epidemics of several previously unreported diseases have occurred and caused severe losses. When elimination, eradication or cultural control is difficult, selective breeding for host resistance to the pathogen may be an attractive option for disease control. However, host resistance is not a panacea and should only be considered when (a) the disease causes severe damage (b) there are no other existing simple cost effective control measures and (c) there is demonstrable genetic variation in resistance and this is not coupled with an excessive level of negative associations with other desirable characteristics. Shrimp have only recently been domesticated and breeding for resistance only began in the mid 1990s; there is limited experience with shrimp breeding in particular and crustaceans in general. Consequently, the principles and concepts behind breeding programmes are based largely on experiences with other species in both the plant and animal kingdoms. Commercial growers now seed ponds with shrimp populations selected for resistance to Taura Syndrome Virus with excellent results, whilst up to now development of White Spot Syndrome Virus resistant populations has been an elusive goal. The original TSV resistant populations were developed using simple mass selection techniques (Colombia). In later generations family based selection has been applied on populations, which initially had survival rates of about 30%, with care taken to reduce inbreeding and loss of genetic variation. This suggests that when the original populations have a reasonable level of resistance, and straightforward, effective selection protocols exist, it is relatively simple to breed for resistance. With catastrophic diseases, such as WSSV, which cause mortalities of 98% or more the frequency of resistance is low and it is suggested that for theoretical reasons single gene, rather than polygenic, resistance is likely to develop. The low frequency of resistance genes in breeding populations may cause genetic bottlenecks which will greatly reduce the genetic variation in the populations. In order to maintain the genetic variation the genes from the small numbers of survivors should be introgressed into populations with broader genetic variability. Furthermore, in order to minimize the probability of breakdown of resistance pyramiding of resistant genes on different loci would be advantageous.Genetic variation in resistance may be encountered either in the initial base populations or may spontaneously arise due to mutations or new recombinants. With extremely prolific species such as shrimps, millions of animals can readily be screened for survival and hence resistant mutants or recombinants may be identified. Once genetic variation has been detected the most appropriate breeding methodology will depend on the nature of both the resistance and the disease or diseases that are of interest to the producers.     

饲料添加益生菌对多病原阳性的凡纳滨对虾生长与存活的影响

采用假交替单胞菌(Pseudoalteromonas sp.) KL-3 2010和微小杆菌(Exiguobacterium sp.) KL-C2 2014作为益生菌,进行凡纳滨对虾(Litopenaeus vannamei)投喂实验,研究上述菌株对带毒对虾的生长与存活的影响。假交替单胞菌KL-3 2010对致急性肝胰腺坏死副溶血弧菌(Vibrio parahemolyticus) (VPAHPND 20130629002S01)有拮抗作用和胞外蛋白酶活性,微小杆菌KL-C2 2014有胞外蛋白酶活性。待试对虾经检测为白斑综合征病毒(WSSV)、致急性肝胰腺坏死病副溶血弧菌(VPAHPND)和虾肝肠胞虫(EHP)弱阳性。经过为期60 d的养殖实验,结果显示,与投喂普通颗粒饲料的对照组相比,投喂假交替单胞菌KL-3 2010的对虾存活率提高了213%±43% (P<0.01);投喂微小杆菌KL-C2 2014的对虾平均生长率提高了105.5%±28.1% (P<0.05);交替投喂2株菌的对虾存活率提高了184%±52% (P<0.05),平均生长率提高了70.6%±32.8%。肠道可培养优势菌研究表明,2株益生菌的投喂显著影响了对虾肠道优势菌群的种类。本研究为带毒虾苗的养殖提供一种有效的病害防控和促生长的手段。     

A stochastic approach for analysis of the influence of white spot disease, zootechnical parameters, water quality, and management factors on the variability of production of shrimp Litopenaeus vannamei cultivated under intensive commercial conditions

We investigated the variability of shrimp Litopenaeus vannamei production by incorporating stochastic elements into deterministic stock models and determined the contribution that white spot disease, zootechnical parameters, water quality, and alternative management strategies have on variability. The model was calibrated for intensive shrimp cultivation in the State of Nayarit, Mexico. Mean annual production increased as a consequence of improved management from 8000 kg ha⁻¹ to 22,000 kg ha⁻¹ when cultivation was not affected by the disease and from 3200 kg ha⁻¹ to 10,400 kg ha⁻¹ when the disease affected production. When simultaneously considering both cases, mean annual production increased from 6300 kg ha⁻¹ to 16,800 kg ha⁻¹. White spot disease was a major factor determining variability of production. Shrimp production was particularly sensitive to levels of dissolved oxygen when management was inadequate, while final weight and mortality rate of shrimp were more sensitive when management improved. Water temperature and salinity had intermediate importance, and mortality caused by the disease and the time when mortality occurred had intermediate or low relevance. Improving management increased shrimp production and diminished variability. The duration of cultivation and stocking density were the most important management variables controlling variability of production when cultivation was affected by the white spot disease. When the disease was not present, pond size and duration of cultivation were the main factors affecting production. Starting time of aeration had relatively lower importance in determining variability, while the stochastic values of dissolved oxygen, in contrast, became most important. These results call for studies on improving aeration management to reduce variability of dissolved oxygen in ponds.     

Light and electron microscopic evidence of white spot disease in the giant tiger shrimp, Penaeus monodon (Fabricius), and the kuruma shrimp, Penaeus japonicus (Bate), cultured in Taiwan

Since 1992, mass mortalities among cultured giant tiger shrimp, Penaeus monodon (Fabricius), and kuruma shrimp, Penaeus japonicus (Bate), have been observed in Taiwan. The condition is known as 'white spot disease' (WSD), based on the characteristic white spots on the cuticle of diseased shrimp. With the scanning electron microscope, two sizes of white spots were observed. Each spot represented a protrusion on the inside surface of the carapace. The composition of white spots was similar to that of the cuticule, most calcium, as determined with an energy dispersive spectrometer. Histological studies of moribund, infected specimens revealed degenerated cells, characterized by hypertrophied nuclei, in various meso- and ectodermal tissues. Infected tissues included cuticular epidermis, connective tissue, lymphoid organ, antennal gland, and haematopoietic, gill and nervous tissue. Nuclei were Feulgen-positive and no occlusion body was found in the necrotic tissue. Transmission electron microscopy revealed the presence of rod-shaped and enveloped virions in the hypertrophied nuclei. The virions measured 298 ± 21 × 107 ± 8 nm in the giant tiger shrimp and 248 ± 12 × 104 ± 8 nm in the kuruma shrimp. In an experimental infection trial, cumulative mortality was 40% within 14 days under stress conditions. No mortality was observed in controls or in non-stressed infected shrimp. Experimental infections show that environmental stressors such as ammonia may enhance the severity of WSD virus infections in cultured shrimp.     

Quantitative genetics of disease resistance in different groups of Penaeus semisulcatus from preliminary controlled challenge test with Vibrio parahaemolyticus the aetiological agent of acute hepatopancreatic necrosis disease (AHPND)

Acute hepatopancreatic necrosis disease (AHPND) has caused losses to shrimp farmers worldwide owing to Vibrio parahaemolyticus (VP_AHPND). Selective breeding for disease resistance of shrimp was used as an alternative strategy for disease control. Five subsamples of shrimp from 20 full-sib families were subjected to a cohabitation challenge test. Juveniles of P. semisulcatus (mean weight 2.5 ± 0.17 g) were tested for survival after challenge with VP_AHPND. Mortality in the challenge test was recorded daily, and the experiment was terminated after 96 days when the mortality ceased. Moderate heritabilities were estimated in resistance to VP_AHPND based on a linear model (LM) and threshold model (TM). This indicates that disease resistance is a heritable trait, and a challenge test may serve as a basis for selection for resistance to VP_AHPND in P. semisulcatus. However, further investigation is needed to confirm and quantify the additive genetic variation for resistance to VP_AHPND in P. semisulcatus.     

Dynamic stock model for analysing semi‐intensive production of whiteleg shrimp Litopenaeus (Penaeus) vannamei affected by the acute hepatopancreatic necrosis disease: Assessment of disease severity indicators and relationships with pond water quality parameters

The production of Litopenaeus vannamei was analysed when affected by the acute hepatopancreatic necrosis disease using a dynamic stock model and primary data of seven production cycles from a shrimp farm in Mexico from 2013 to 2016. Significant results (p < .05) of the correlation analysis indicated that during those years mortalities by the disease were more severe when water salinity was high and productivity was low. Significant results from ANOVA showed that throughout the period, disease severity and salinity diminished while pond productivity initially declined but subsequently improved. Significant results from regression analyses conducted for each production cycle also indicated the importance of salinity and productivity on disease severity and showed that early mortality by the disease occurred in ponds with warmer water. Within the observed range of water quality parameters, increases of 1 cm in water transparency and 1 g/L in salinity resulted in increments within 0.17%–0.25% and 1.7%–3.1% in shrimp mortality by the disease. When increases of 1°C in water temperature were recorded, outbreaks occurred 0.2–1.57 weeks earlier. In conclusion, the disease strongly determines the dynamics of shrimp production, and the role of salinity, productivity and temperature is worthy of further delving.     

Identification of RAPD‐SCAR marker linked to white spot syndrome virus resistance in populations of giant black tiger shrimp,Penaeus monodon Fabricius

White spot disease (WSD) caused by white spot syndrome virus (WSSV) creates severe epizootics in shrimp aquaculture industry worldwide. Despite several efforts, no such permanent remedy was yet developed. Selective breeding using DNA markers would be a cost‐effective strategy for long‐term solution of this problem. In the present investigation, out of 30 random primers, only one primer produced a statistically significant (P < 0.01) randomly amplified polymorphic DNA (RAPD) marker of 502 bp, which provided a good discrimination between disease resistant and disease susceptible populations of Penaeus monodon from three geographical locations along the East coast of India. Because RAPD markers are dominant, a sequence characterized amplified region (SCAR) marker was developed by cloning and sequencing of 502 bp RAPD fragment, which generates a single 457 bp DNA fragment after PCR amplification only in the disease resistant shrimps. Challenge experiment was also conducted to validate this 457 bp SCAR marker, and the results suggested that the WSSV loads were 2.25 × 10³ fold higher in disease susceptible than that in disease resistant shrimps using real‐time PCR. Therefore, this 457 bp DNA SCAR marker will be very valuable towards the development of disease‐free shrimp aquaculture industry.     

The first record of acute hepatopancreatic necrosis disease in the Philippines

A new and emerging disease is threatening the shrimp industry, a bacterial disease which contains a highly pathogenic plasmid, creating a deadly toxin that causes high mortality in shrimps. The disease has been identified as acute hepatopancreatic necrosis disease (AHPND) or commonly known as early mortality syndrome (EMS). To help the efforts of sustaining the shrimp industry, the study focused on detecting Vibrio parahaemolyticus causing AHPND/EMS affecting Penaeus vannamei (Boone, 1931) (Pacific white shrimp) and Penaeus monodon (Fabricius 1798) (Black Tiger shrimp) in different locations in the Philippines. The presence of the disease was determined by microbiological methods and polymerase chain reaction (PCR) and was confirmed by the histopathology of the hepatopancreas of infected shrimp. Results show that the prevalence of the pathogenic strain of V. parahaemolyticus from the shrimps collected were 33% in Luzon, 21% in Visayas and 5% in Mindanao. The study presents the first record of AHPND/EMS in the country. The detection of this newly emerging disease in the shrimp industry is very crucial as it is the first step in identifying affected sites towards strategizing ways to combat the disease.     

Use of GIS and machine learning to predict disease in shrimp farmed on the east coast of the Mekong Delta,Vietnam

Diseases in shrimp farms in the Mekong Delta of Vietnam cause significant crop losses and are therefore of great concern to producers. Once a pond becomes infected, it is difficult to prevent spread of the disease to nearby shrimp farming areas. Thus, predicting the occurrence of disease is an essential part of reducing the risk for shrimp farmers. In this study, we applied an integrated geographic information system and machine learning system to predict three serious diseases of shrimp, namely, acute hepatopancreatic necrosis, white spot syndrome disease, and Enterocytozoon hepatopenaei infection, based on data collected from shrimp farms in the Tra Vinh, Bac Lieu, Soc Trang, and Ca Mau provinces of Vietnam. We first constructed a map showing the distribution of these diseases using the locations of affected farms, and then we conducted spatial analysis to acquire the geographical features of the affected locations. This latter information was combined with environmental factors and clinical signs to form the set of independent variables affecting the outbreak of diseases. The neural network model outperformed the logistic regression, random forest, and gradient boosting methods in terms of predicting infection to estimate the probability of disease occurrence in farmed areas. Acute hepatopancreatic necrosis disease infected farms downstream of the Co Chien and Hau Rivers of Tra Vinh and west of Ca Mau. Enterocytozoon hepatopenaei infection is distributed in Soc Trang Province, while white spot syndrome virus has spread to the coastal districts of Soc Trang and Bac Lieu Provinces, where it is highly associated to water from a complex canal system.

     

Effect of Water Temperature on Reproductive Tract Condition of Penaeus setiferus Adult Males

Abstract— This study was conducted to determine the effect of water temperature on male reproductive tract degenerative syndrome (MRTDS) and male reproductive system melanization (MRSM) in Penaeus setiferus , and to evaluate the effect of water temperature on spermatophore regeneration time and sperm quality in electrically ejaculated adult males. Variation in sperm quality, gonadosomatic index, and histological changes in the reproductive system were used to assess reproductive tract changes in the first experiment. Sperm quality and regeneration time were measured before and after successive regenerations in the second experiment. Sperm quality was stable when shrimp were held at 26 C for 30 d but was reduced when shrimp were held at 30 C. Reproductive tissues of 20% of the shrimp held at 30 C were melanized to some degree but tissue melanization was found in only 2.5% of the shrimp held at 26 C. Spennatophore regeneration time was also affected by temperature. Average times for first spermatophore regeneration were 192 h at 25 C, 152 h at 30 C, and 144 h at 33 C. Sperm quality of regenerated shrimp was not affected at 25 C but was reduced for regenerated animals held at 33 C. Based on these results, we recommend a water temperature between 25 and 27 C to obtain adequate spermatophore regeneration and to maintain healthy male P. setiferus broodstock for at least 30 d.     

蚯蚓与蝇蛆对中国对虾生长及抗白斑综合征病毒感染的研究

以中国对虾（Fenneropenaeus chinensis）为实验材料,分别投喂配合饲料、蚯蚓、蛤蜊、蝇蛆等四种饵料,利用生长和抗病性指标综合评价蚯蚓和蝇蛆作为中国对虾饵料的可行性。生长实验前测定每个实验组的初始体长和体重,养殖40d后再次测定生长指标,之后,分别投喂不同毒饵量进行人工感染实验。方差分析表明：投喂四种饵料后,蛤蜊组生长最快,其次是蚯蚓,再次是蝇蛆,最后是配合饲料,各组之间对虾体长增长差异显著（P<0.05）,体重增长差异极显著(P<0.01) 。利用线性固定模型分析不同饵料及不同病毒量感染下对中国对虾存活率的影响,其中蚯蚓组存活率最高,配合饲料组存活率最低,并且蚯蚓组存活率显著高于蛤蜊组和配合饲料组(P<0.05),蝇蛆组显著高于配合饲料组（P<0.05）。研究结果表明：蚯蚓和蝇蛆可显著提高中国对虾的生长速度,明显提高中国对虾的抗病性,是很好的对虾饵料。     

罗氏沼虾开放和闭锁核心育种群生长和存活性能分析

开放核心育种体系可增加核心群体的选择强度,进而增加育种目标性状的遗传进展。本研究以罗氏沼虾(Macrobrachium rosenbergii)为研究对象,向闭锁核心群(closed nucleus population,NP)内引入扩繁群(multiplier population, MP)个体,构建两个杂交群体(正交群体NP/MP,反交群体MP/NP),并以闭锁核心群(NP/NP)为对照,应用线性混合效应模型(linear mixed effects model, LME)和广义线性混合模型(generalized linear mixed model, GLMM)对不同群体收获体重和存活率的估计边际均值进行比较,评估在虾类选择育种中构建开放核心育种体系的可行性。结果显示:(1) NP/NP、MP/NP和NP/MP群体的收获体重估计边际均值分别为45.83 g、49.57 g和46.62 g;与NP/NP群体相比, MP/NP和NP/MP群体分别提高了8.16%和1.72%。(2) NP/NP、MP/NP和NP/MP群体的存活率估计边际均值分别为72.92%、68.04%和66.55%;与NP/NP群体相比,MP/NP和NP/MP群体分别降低了6.69%和8.74%。综上所述,在罗氏沼虾核心群中引入扩繁群个体,构建开放核心育种群体,可以有效地增加收获体重的遗传进展;同时在制定选择指数时,应加大存活性状的权重,选择生长和存活性能均优良的家系生产扩繁群体,导入闭锁核心育种群,构建生长和存活性能均优良的开放核心育种群。     

一种复合益生菌对凡纳滨对虾抗副溶血弧菌感染能力的影响

本研究选择1株地衣芽孢杆菌(Bacillus licheniformis)BL-9、1株枯草芽孢杆菌(Bacillus subtilis)BS-12、1株金丽假交替单胞菌(Pseudoalteromonas flavipulchra)CDM8,以1∶1∶1将其组成复合益生菌,各益生菌水体添加浓度为107 CFU/ml,进行为期30 d的凡纳滨对虾(Litopenaeus vannamei)养殖实验。实验分为暂养期(7d)、益生菌处理期(15d)、副溶血弧菌(Vibrio parahaemolyticus)攻毒期(10 d)。结果显示,养殖水体中添加复合益生菌能显著增加水体和对虾肠道可培养细菌总数(P<0.05),攻毒实验结束时,实验组对虾累积存活率为(73.33±6.83)%,显著高于阳性对照组(25.33±15.43)%。对虾抗病基因热激蛋白70(Heat shock proteins 70,Hsp70)、β-1,3-葡聚糖结合蛋白-脂蛋白(Beta-1,3-glucan-binding protein-lipoprotein,βGBP-HDL)、脂多糖-β-1,3-葡聚糖结合蛋白(Lipopolysaccharide-β-1,3-glucan binding protein,LGBP)、抗菌肽Crustin在益生菌处理阶段均出现不同程度的上调,在攻毒阶段虽呈现各自不同的表达情况,但所有基因都经历了更大幅度上调。研究表明,水体中添加芽孢杆菌和假交替单胞菌组成的复合益生菌可提高凡纳滨对虾抗副溶血弧菌感染能力,对虾抗病力的提高可能与益生菌增加对虾肠道可培养细菌数量、抗病相关基因表达水平及过氧化氢酶(CAT)活性有关。     

DEVELOPMENT OF AN INTENSIVE SHRIMP CULTURE SYSTEM IN KUWAIT1

Development of an intensive culture system is presently being conducted in Kuwait as a means of farming penaeid shrimp in arid lands. Efficiency of commercial-scale hatchery production of Penaeus semisulcatus and P. japonicus showed significant improvement in 1981 over previous years. Three one-month hatchery rearing cycles in three 15 m³ concrete tanks yielded a total of 9.1 million post-larval shrimp with an average density of 160 PL/liter being obtained for one of these trials. Four other trials were discontinued due to low spawning rates and disease. Installation of a heat exchange system made it possible to begin larviculture in February, two months earlier than in previous years. Research in nursery and grow-out phases of intensive shrimp culture is being directed towards raceways, although Shigueno culture systems are also being investigated. Experimental work with six 18 m × 1.5 m raceways demonstrated that low water exchange rates (1 tank volume/day), combined with high aeration (9.5 liter/min) yielded the highest shrimp biomass. Unheated greenhouse structures constructed over the raceways showed that a single layer of 0.25 mm clear plastic sheeting could maintain water temperatures up to 2.7°C above those in unenclosed tanks. Rearing trials in two Shigueno tank systems demonstrated the importance of high water flow rates (3 tank volumes/day minimum) and adequate aeration. Due to sub-optimal conditions, survival was reduced to 7.5%, however, growth rates up to 6 g/month were obtained for P. semisulcatus.     

Concurrent rice‐shrimp‐crab farming systems in the Mekong Delta: Are conditions (sub) optimal for crop production and survival?

The Mekong Delta is the most important rice‐ and shrimp‐producing region for food and economic security in Vietnam. Rice‐shrimp farming is practised where salinity fluctuates substantially between wet and dry seasons. Research points to several potential risk factors for rotational systems, but how these link directly to both rice and shrimp production remains poorly quantified for systems that stock and harvest animals year‐round. We examined water and soil quality of 18 rice‐shrimp‐crab ponds, in which shrimp and crab are grown in both wet and dry seasons, in the Cà Mau Province of Vietnam. Multiple lines of evidence indicated that environmental conditions experienced by both rice and shrimp were suboptimal and contributed to low yields and survival. Year‐round cropping of shrimp and crab was associated with sustained suboptimal salinity, intensified by drought, for the wet‐season cultivation of rice. Although rice seedlings were sown in all 18 ponds, only three had a harvestable crop. Low shrimp production and survival was associated with sustained suboptimal water temperatures (too high), salinity (too high in the dry season and too low in the wet season) and dissolved oxygen concentrations (too low). Food availability and quality may also have affected shrimp production. Improving productivity of rice‐shrimp‐crab ponds in the study region may require (1) separation of rice and shrimp crops and improving efficiency of soil washing practices such that salinity conditions are more suitable for each when grown, and (2) management intervention to increase oxygenation of water, and the availability and quality of food for shrimp.     

Response of Penaeus vannamei to Dietary Calcium, Phosphorus and Calcium: Phosphorus Ratio

Two experiments, consisting of 7 d conditioning periods and 33 d feeding trials, were conducted to evaluate the response of juvenile Penaeus vannamei to dietary calcium, phosphorus and the effects of the calcium to phosphorus ratio on growth and tissue mineralization. In both experiments, the main effects, calcium and phosphorus, and their interaction significantly affected growth and survival of the shrimp. In the absence of supplemental calcium, the basal diet containing 0.35% phosphorus was adequate to maintain good growth and survival of shrimp. In the presence of replete phosphorus, supplementation of 1.0 and 2.0% calcium to the diet depressed survival and did not appear to increase the nutritive value of the diet, indicating that a dietary calcium supplement is not required under these conditions. In the presence of 1.0 and 2.0% supplemental calcium, supplementation of 0.5–1.0% phosphorus and 1.0–2.0% phosphorus, respectively, were required to maintain normal growth of the shrimp. At high levels of calcium supplementation, poor growth was observed at the levels of phosphorus supplementation evaluated. Although the calcium and phosphorus content of the hepatopancreas and the carapace responded to dietary supplementation, there was no clear correlation of tissue mineralization to shrimp growth.     

A Microsatellite DNA Marker Developed for Identifying Disease‐resistant Population of Giant Black Tiger Shrimp,Penaeus monodon

White spot disease caused by white spot syndrome virus (WSSV) poses major problems that result in huge economic losses each year in shrimp aquaculture throughout the world. In the present study, microsatellite‐based DNA fingerprints have been compared between naturally occurring WSSV disease‐resistant and susceptible populations of giant black tiger shrimp, Penaeus monodon, to find DNA markers. For the first time, we report here a microsatellite locus, which, after amplification by polymerase chain reaction, provides a highly statistically significant DNA fingerprint of 71 bp, only in disease susceptible populations but not in disease‐resistant shrimp populations, whereas a 317 bp band is common in both. The absence of the former DNA marker will be very useful to identify disease‐resistant broodstock of P. monodon for marker‐assisted selection in breeding programs to generate disease‐free shrimps (P. monodon) in the aquaculture industry.     

Bath efficacy of sodium hypochlorite,oxytetracycline dihydrate and chloramphenicol against bacterial black disease in fairy shrimp Branchinella thailandensis

To control black disease infecting fairy shrimp Branchinella thailandensis, the effects of concentrations and exposure time to three effective antimicrobials, which inhibited the pathogens in vitro, were evaluated. Exposure to sodium hypochlorite (NaOCl) caused a great toxicological response in the shrimp, 100% mortality was observed within 30 min–2 h at 5–20 μg mL^?1. For oxytetracycline dihydrate (OTC) and chloramphenicol (CP), short‐term exposure to four high concentrations up to 5 h and long‐term exposure (12 days) to four low concentrations were used to determine an appropriate method for bath efficacy. Long‐term exposure to low concentrations was more toxic than the short‐term. Short‐term exposure to OTC showed the highest survival rate and CP was considered more toxic. The minimum survival rate of the shrimp exposed to both antibiotics at 250 μg mL^?1 for 3 h was 83.3%. For determination of the bath efficacy, a short–term exposure (3 h) to OTC and CP was conducted using artificially infected shrimp. Administration of OTC and CP at 250 and 500 μg mL^?1 resulted in the highest survival rates of 56.7% and 46.7% respectively. This study demonstrated that bath administration with OTC could be an alternative method for the treatment of black disease in fairy shrimp cultivation.