Inbreeding and disease in tropical shrimp aquaculture: a reappraisal and caution

The disease crisis facing shrimp aquaculture may be propelled, in part, by an interaction between management practices that cause inbreeding, and the amplification by inbreeding of susceptibility to disease and environmental stresses. The study describes and numerically simulates gene flow from Penaeus (Litopenaeus) vannamei hatcheries that employ a ‘Breeder Lock’ to discourage use of their PL as breeders, through ‘copy hatcheries’ that breed the locked PL, to inbred shrimp in farm ponds. Re‐analysis of published data shows that inbreeding depression under stress is exceptionally strong in shrimp. Inbreeding is currently overlooked as a problem because: (1) procedures recommended for well‐managed hatcheries do not consider their implications for the copy hatcheries that supply most farmed shrimp (estimated 70%), (2) inbreeding in hatcheries is often reported as zero even though zero is the mathematical expectation of the usual estimator (Fis, fixation index) whatever the true genealogy of the broodstock. Simulation shows, however, that inbreeding can be estimated with Wang's trioML estimator, that Fis can differentiate Breeder Locked from copy PL and that simple tests can verify the lock status of PL. The importance of inbreeding should be re‐evaluated in the context of disease and environmental stress. Unrecognized inbreeding may increase the incidence, prevalence and lethality of WSSV, IHHNV, EMS (AHPND) and other diseases.     

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.     

Update on early mortality syndrome/acute hepatopancreatic necrosis disease by April 2018

Outbreaks of acute hepatopancreatic necrosis disease (AHPND) have caused great economic losses to many shrimp‐producing countries in Asia since its first appearance in 2009. The causative agent was reported in 2013 as specific isolates of Vibrio parahaemolyticus (VP_AHPND) that were later found to harbor a plasmid (pVA) encoding the Pir‐like binary toxin genes Pir ^vpA and Pir ^vpB. VP_AHPND isolates colonize the shrimp stomach and release the binary toxins that cause massive sloughing of tubule epithelial cells followed by shrimp mortality. More recent information indicates that pVA plasmid and variants occur in many V. parahaemolyticus serotypes and also in other Vibrio species such as Vibrio campbellii, Vibrio harveyi, and Vibrio owensii. Information on such genomic and proteomic studies of different VP_AHPND isolates from different countries are reviewed. A cohort study carried out in Thailand in 2014 indicated that AHPND outbreaks account for only a portion of the disease outbreaks reported by shrimp farmers as outbreaks of early mortality syndrome (EMS). It is recommended that a regional research network and surveillance program for newly emerging or re‐emerging pathogens be established to speed up the process of diagnosis and the implementation of coordinated control measures and to avoid a repeat of the EMS/AHPND scenario.     

对虾养殖场底泥中副溶血性弧菌的接合型质粒多样性及其与毒力之间的关系

为了阐明引起急性肝胰腺坏死病(acute hepatopancreatic necrosis disease, AHPND)副溶血性弧菌的接合型质粒在对虾养殖环境中的遗传多样性,实验从中国5个沿海省份的虾场收集了100个底泥样品,以质粒上编码接合转移蛋白的保守基因为目标,利用PCR法检测相关质粒的存在情况,并对质粒进行测序。结果显示,100个样品中有39个样品含有质粒的接合转移蛋白片段。从100个底泥样品中分离出15株副溶血性弧菌,其中13株含有1～2个质粒。质粒序列测序结果显示,这些质粒可分为8种类型/谱型,其中7种不携带pirAB,但均含有编码接合转移的基因簇。根据分离副溶血性弧菌携带质粒的8种谱型,分别选择8株副溶血性弧菌进行凡纳滨对虾攻毒实验,发现这些菌株对凡纳滨对虾的毒性有显著差异,实验虾死亡率为15%～100%。只有pirAB阳性菌株会对实验虾产生AHPND症状,死亡率为100%。对质粒组成进行分析表明,质粒之间遗传物质交换频繁,大部分质粒的遗传组成都来自一个183 kb的超大质粒pVP2HP。综上,本实验通过探究对虾养殖场底泥中结合性质粒的多样性,增强了人们对副溶血性弧菌...     

Characterization of Vibrio parahaemolyticus causing acute hepatopancreatic necrosis disease in southern Thailand

Vibrio parahaemolyticus was isolated from shrimp of five farms located in the Pattani and Songkhla provinces of southern Thailand. Using a PCR method targeted to the unique DNA sequences derived from the plasmid (AP2 primers) and the toxin gene (AP3 primers) of V. parahaemolyticus that caused acute hepatopancreatic necrosis disease (AHPND), a total of 33 of 108 isolates were positive. In contrast, all 63 and 66 isolates of clinical and environmental V. parahaemolyticus, respectively, obtained previously from 2008 to 2014 from the same area were negative. This implied that these strains were likely to be the cause of the outbreak of AHPND in this area. Intestinal samples proved to be a better source for the isolation of V. parahaemolyticus AHPND than the hepatopancreas. All isolates were investigated for haemolytic activity, virulence genes, serotypes, genotypes and antibiotic susceptibility. All the AHPND isolates had a unique O antigen, but small variations of the K antigens were detected from different farms. In addition, the DNA profiles of V. parahaemolyticus AHPND isolates were similar, but distinct from those clinical and environmental isolates. It is postulated that the causative agent of AHPND might have originated from one clone and then slightly different serotypes subsequently developed.     

Studies on the occurrence of infectious myonecrosis virus in pond‐reared Litopenaeus vannamei (Boone, 1931) in India

Whiteleg shrimp, Litopenaeus vannamei, with clinical sign of muscle opaqueness with reddish colour at the distal abdominal segments were observed in farms located in West Bengal State, India. The mortality of shrimp in all disease outbreak ponds ranged from 20% to 50%, and mortality increased gradually. The RT‐PCR assay of these samples using primer sets specific to infectious myonecrosis virus (IMNV) revealed its presence in the disease outbreak ponds. The IMNV infection was reproduced in healthy shrimp by intramuscular injection to satisfy River's postulates. The virus caused mortality in intramuscularly challenged shrimp, but failed to cause mortality by oral route. Tissue distribution of IMNV in infected shrimp by RT‐PCR assay revealed the presence of this virus in haemolymph, gill, hepatopancreas and muscle. This study confirms that the disease outbreak which occurred in the shrimp farms located at Purba Medinipur District, West Bengal, India, was due to IMNV.     

Identification of potential general markers of disease resistance in Exopalaemon carinicauda via three‐round challenge selection with an acute hepatopancreatic necrosis disease‐causing strain of Vibrio parahaemolyticus

Sixteen candidate disease‐resistant parameters were selected through which to evaluate the acute hepatopancreatic necrosis disease (AHPND)‐resistant capability of Exopalaemon carinicauda after three generations of selection for AHPND‐causing strain of Vibrio parahaemolyticus (VP_AHPND) resistance in our previous study. However, these parameters required further verification. In this study, another AHPND‐resistant E. carinicauda series was obtained through a short‐term selection procedure, consisting of three virulent challenge rounds of selection (about three‐week interval for each challenge) with VP_AHPND infection. After this selection, the survival rate at 144 hr post infection (hpi) increased from 23.33% to 37.78% and the observed 48‐hr LD₅₀ of VP_AHPND to shrimp increased from 10^5.5 cfu/ml to 10^6.5 cfu/ml. Then, the immune response of this AHPND‐resistant E. carinicauda was studied using the 16 candidate AHPND‐resistant parameters selected for in our previous study. The improved VP_AHPND clearance rate in hpi, increased total haemocyte counts, haemocyanin concentration, alkaline phosphatase activity and expressions of six immune‐related genes (Tollip and ALF in haemocytes and hepatopancreas; lysozyme, crustin and cathepsin B in hepatopancreas; and LGBP in haemocytes) at 24 hpi after the three‐round challenge selection suggest that these immune parameters may be reliable markers for the evaluation of the physiological status and potential AHPND‐resistant phenotypes in E. carinicauda.     

Anti‐PirA‐like toxin immunoglobulin (IgY) in feeds passively immunizes shrimp against acute hepatopancreatic necrosis disease

Acute hepatopancreatic necrosis disease (AHPND), caused by a toxin‐producing Vibrio parahaemolyticus strain, has become a serious threat to shrimp aquaculture. The need to regulate antibiotic use prompted the development of alternative ways to treat infections in aquaculture including the use of chicken egg yolk immunoglobulin (IgY) for passive immunization. This study evaluated the protective effect of IgY against AHPND infection in Litopenaeus vannamei (Boone). IgY was isolated from eggs laid by hens immunized with recombinant PirA‐like (rPirA) and PirB‐like (rPirB) toxins. Whole‐egg powders having IgY specific to rPirA (anti‐PirA‐IgY) and rPirB (anti‐PirB‐IgY) and IgY from non‐immunized hen (control‐IgY) were mixed with basal diets at 20% concentrations and used to prefeed shrimp 3 days before the bacterial challenge test. Survival rates of the challenged shrimp fed the anti‐PirA‐IgY, anti‐PirB‐IgY and control‐IgY diets were 86%, 14% and 0%, respectively. Only the feed containing anti‐PirA‐IgY protected shrimp against AHPND. Increasing the concentration of rPirA antigen to immunize hens and lowering the amount of egg powder in feeds to 10% consistently showed higher survival rates in shrimp fed with anti‐PirA‐IgY (87%) compared with the control (12%). These results confirm that addition of anti‐PirA‐IgY in feeds could be an effective prophylactic method against AHPND infection in shrimp.     

Comparing Profits from Shrimp Aquaculture with and without Green-Water Technology in the Philippines

To reduce disease impact, Philippine farmers developed the green-water (GW) system, which has been spreading rapidly since 2008. In the most applied GW-system, the shrimp pond receives water from a reservoir stocked with tilapia. We collected financial data on GW and non-GW systems from farms having similar management and environmental context. All farms had more than one pond; the average pond area was < 1 ha, and total pond areas per farm ranged from 1.4 to 139 ha. The total variable and fixed costs per ha of shrimp pond were not different for GW and non-GW farms, but on GW farms the cost per kg of shrimp produced was lower. The latter was higher than the 2009 market price of 30 g shrimp for non-GW farms mainly. In GW farms the cost of seed was lower, and survival rates and individual shrimp weights at harvest were higher for an equally long culture period.     

“Summer syndrome” in Litopenaeus stylirostris grow out ponds in New Caledonia: Zootechnical and environmental factors

The “Summer Syndrome”, a septicemic vibriosis caused by Vibrio nigripulchritudo, is responsible for one of the two main seasonal mortalities which affect shrimp aquaculture in New Caledonia. It was identified for the first time in December 1997 in an intensive shrimp farm (called farm DF) and has been enzootic ever since. The “Summer syndrome” affects shrimp during warm-season growout. Although the geographic area concerned is limited, it is a potential threat for the industry in that the disease could spread. Analysis of grow out data from 1991 to 2002 in this farm was carried out in order to characterize the disease and the environmental conditions in relation to mortality. Results of farm DF were compared to data from farms not affected by the disease (called farms HC). The “Summer syndrome” occurs during the warm season which is characterized by an increase in water temperature and sunny duration. This disease is characterized by simultaneously highest densities and weakest drying duration between crops in the farms studied. Between 1991 and 2002, the increase of feed input and nitrogen rate in feed has led to a significant increase of the growth rate over the years in farm DF. At the same time, water management has not changed and has induced an increase in environmental waste production. In consequence, an early eutrophication of water is observed, which is concomitant with the beginning of the mortality outbreaks. This could play a role by inducing directly or indirectly a stress for shrimp and/or a growth and/or virulence factors of the pathogen.     

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.

     

Biochemical changes and tissue distribution of Enterocytozoon hepatopenaei (EHP) in naturally and experimentally EHP‐infected whiteleg shrimp,Litopenaeus vannamei (Boone, 1931), in India

Stunted growth in pond‐reared Litopenaeus vannamei was observed in different farms located in Tamil Nadu and Andhra Pradesh, India. No mortality was associated with stunted growth. PCR assay on these samples revealed the presence of Enterocytozoon hepatopenaei (EHP) in stunted shrimp. Tissue distribution of EHP in naturally and experimentally infected shrimp was studied by PCR and histology. Histological examination revealed the presence of EHP in hepatopancreas and gut, but not in other organs. The PCR assay revealed the presence of EHP in all the organs tested in both naturally and experimentally infected shrimp. Healthy shrimp were challenged with E. hepatopenaei by intramuscular injection and oral route, and no mortality was observed in both routes after 30 days post‐challenge. Different developmental stages of the microsporidian parasite were observed in the hepatopancreatic epithelial cells. Biochemical parameters such as total protein, albumin, aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase were measured in the haemolymph of naturally and experimentally EHP‐infected shrimp. All biochemical parameters mentioned were found to be significantly higher in EHP‐infected shrimp when compared to normal shrimp. This is the first report relating AST and ALT levels to EHP infection in naturally and experimentally infected shrimp.     

Influence of variation in water temperature on survival,growth and yield of Pacific white shrimp Litopenaeus vannamei in inland ponds for low‐salinity culture

There is considerable interest in the culture of whiteleg shrimp (Litopenaeus vannamei) in inland low‐salinity water in Alabama and other states in the Sunbelt region of the US. However, the growing season is truncated as compared with tropical or subtropical areas where this species is typically cultured, and temperature is thought to be a major factor influencing shrimp production in the US. This study, conducted at Greene Prairie Aquafarm located in west‐central Alabama, considered water temperature patterns on a shrimp farm in different ponds and different years; and sought possible effects of bottom water temperature in ponds on variation in shrimp survival, growth and production. Water temperature at 1.2 m depth in 22 ponds and air temperature were monitored at 1‐hr intervals during the 2012, 2013, 2014 and 2015 growing seasons. Records of stocking rates, survival rates and production were provided by the farm owner. Correlation analysis and linear mixed model analysis of variance were used. Results showed that hourly water temperatures differed among ponds. The range of water temperature in each pond explained 41% of the variance in average final weight of shrimp harvested from each pond. In conclusion, the results suggest that variation in water temperature patterns has considerable influence on shrimp growth and survival in ponds.     

Polyclonal antibody‐based farmer‐friendly flow‐through test for the detection of acute hepatopancreatic necrosis disease in shrimp

Early mortality syndrome (EMS) or acute hepatopancreatic necrosis disease (AHPND) is currently the most significant disease of shrimp in farms of Vietnam, Thailand, Malaysia, China and Mexico, and there is a great risk that it may spread to other shrimp farming countries. Although, an array of sophisticated detection tools for AHPND available, there is a need for a sensitive, simple and rapid detection method. In this study, a simple, sensitive, rapid and polyclonal antibody‐based farmer‐friendly flow‐through assay (FTA) test has been developed for the detection of AHPND pathogen. The recombinant Photorhabdus insect‐related (Pir) A toxin‐like protein of AHPND pathogen was used to immunize rabbits at 21‐day interval observed for highest antibody titre after third booster by ELISA. The raised rabbit antiserum was purified by affinity chromatography and characterized by Western blot. The antiserum showed no cross‐reactivity with AHPND‐free Vibrio parahaemolyticus, V. anguillarum, White Spot Virus (WSV), Aeromonashydrophila and Aphanomycesinvadans. This polyclonal rabbit antiserum was used to develop a farmer‐friendly FTA test for the detection of AHPND pathogen. This simple FTA testis is more sensitive and could detect PirA^VP toxin up to 0.121 µg/ml, compared with 0.242 µg/ml by immunodot assay. Furthermore, FTA test requires only 8–10 min for completion, compared with 3 hr by immunodot thus found to be more sensitive, specific and cost‐effective. Collectively, sensitive FTA test would help shrimp farmers to take real‐time management decisions, especially emergency harvest and finally be a better hope for the prevention of AHPND.     

Economic characteristics of extensive shrimp farms in the Mekong delta

Abstract

Extensive forms of shrimp aquaculture have become an important source of income for farm households in the brackish water region of the Mekong Delta, Vietnam. The economic and production characteristics of farms are discussed in this paper, based on a 1997 farm survey, with particular emphasis on shrimp farming practices. Two types of shrimp fanning practice were observed, one based on natural recruitment of seedstock with few supplementary inputs, and the other based on relatively high cash investments in Penaeus monodon seedstock and other inputs. Households practising monodon culture made significantly more income, but faced high risk associated with shrimp mortality. An investigation of the factors affecting monodon yields indicated that investment in higher quality inputs had a positive effect on yield and income. However, further research is needed on the extent to which management can reduce risk of shrimp mortality by investing in such premium quality inputs.     

White spot syndrome virus epizootic in cultured Pacific white shrimp Litopenaeus vannamei (Boone) in Taiwan

White spot syndrome virus (WSSV) has caused significant losses in shrimp farms worldwide. Between 2004 and 2006, Pacific white shrimp Litopenaeus vannamei (Boone) were collected from 220 farms in Taiwan to determine the prevalence and impact of WSSV infection on the shrimp farm industry. Polymerase chain reaction (PCR) analysis detected WSSV in shrimp from 26% of farms. Juvenile shrimp farms had the highest infection levels (38%; 19/50 farms) and brooder shrimp farms had the lowest (5%; one of 20 farms). The average extent of infection at each farm was as follows for WSSV‐positive farms: post‐larvae farms, 71%; juvenile farms, 61%; subadult farms, 62%; adult farms, 49%; and brooder farms, 40%. Characteristic white spots, hypertrophied nuclei and basophilic viral inclusion bodies were found in the epithelia of gills and tail fans, appendages, cephalothorax and hepatopancreas, and virions of WSSV were observed. Of shrimp that had WSSV lesions, 100% had lesions on the cephalothorax, 96% in gills and tail fans, 91% on appendages and 17% in the hepatopancreas. WSSV was also detected in copepoda and crustaceans from the shrimp farms. Sequence comparison using the pms146 gene fragment of WSSV showed that isolates from the farms had 99.7–100% nucleotide sequence identity with four strains in the GenBank database – China ( AF332093 ), Taiwan ( AF440570 and U50923 ) and Thailand ( AF369029 ). This is the first broad study of WSSV infection in L. vannamei in Taiwan.     

Economics of Sustainable Intensification of Aquaculture: Evidence from Shrimp Farms in Vietnam and Thailand

There is growing interest in sustainable intensification of aquaculture production. Yet little economic analysis has been done on farm‐level effects of the economic sustainability of production intensification. Data from 83 shrimp farms (43 in Vietnam and 40 in Thailand) were used to identify (through principal component and cluster analyses) 13 clusters of management practices that reflected various scales of production intensity that ranged from 0–1999 kg/ha/crop to 10,000 kg/ha/crop and above, for both Penaeus monodon and Litopenaeus vannamei in Vietnam and Thailand. The clusters identified reflected sets of management practices that resulted in differing yields despite similarities in stocking densities among some clusters. The enterprise budget analysis developed showed that the more intensively managed clusters outperformed the less intensively managed clusters in economic terms. More intensively managed farm clusters had lower costs per metric ton of shrimp produced and were more profitable. The greater yields of shrimp produced per hectare of land and water resources in more intensively managed shrimp farms spread annual fixed costs across a greater volume of shrimp produced and reduced the cost per metric ton of shrimp. Costs per metric ton of shrimp produced decreased from the lowest to the highest intensity level (from US$10,245 at lowest intensity to US$3484 at highest for P. monodon and from US$24,301 to US$5387 for L. vannamei in Vietnam and from US$8184 at the lowest intensity level to US$3817 at the highest intensity level per metric ton for L. vannamei in Thailand). Costs of pond amendments used in shrimp production were particularly high in Vietnam and largely unwarranted, whereas fixed costs associated with the value of land, production facilities, equipment, and labor were sufficiently high in Thailand so that net returns were negative in the long run. Nevertheless, economic losses in Thailand were less at greater levels of intensification. The study demonstrated a clear value proposition for shrimp farmers to use natural resources (such as land) and other inputs in an efficient manner and supports findings from corresponding research on farm‐level natural resource use efficiency. Additional research that incorporates economic analysis into on‐farm studies of sustainable intensification of aquaculture is needed to provide ongoing guidance related to sustainable management practices for aquaculture.     

Progress in research on acute hepatopancreatic necrosis disease (AHPND)

Acute hepatopancreatic necrosis disease (AHPND) of shrimps is an important disease, first appeared in China in 2009. Since then, AHPND has caused serious drops in shrimp production (up to 20 % worldwide). Although AHPND [originally termed as acute hepatopancreatic necrosis syndrome (AHPNS)] first appeared in 2009, it was not until 2013 that a laboratory infection model was devised and the causative agent identified as certain strains of Vibrio parahaemolyticus. AHPND has caused mortality from 40 to 100 % which usually occurs early (within approximately 35 days) after stocking shrimp fry in shrimp ponds; therefore, it was initially referred to as early mortality syndrome (EMS). Confusingly, other pathogens and environmental factors also cause EMS and are often attributed to AHPND by shrimp farmers. Frequently, farmers do not send samples for confirmatory tests requiring detection of the unique histopathology at the acute stage of disease (massive sloughing of hepatopancreatic epithelial cells without any accompanying signs of a pathogen). The gross signs presumptive of AHPND (lethargy, slow growth, empty stomach and midgut, and a pale to white, atrophied hepatopancreas) are insufficient for confirmatory diagnosis. Recently, molecular detection of AHPND bacteria using PCR has been developed, which has sped up diagnosis and increased research on the causative agent, alternative detection methods, and possible therapies. We hope that this review of research progress on AHPND will serve as a useful introduction for researchers who are currently unfamiliar with AHPND, but have backgrounds in bacterial virulence, detection, and epidemiology, and may be encouraged to participate in the research effort to reduce AHPND’s impact on shrimp cultivation.     

Pond-to-pond variability in post-larval shrimp, Litopenaeus vannamei, survival and growth in inland low-salinity waters of west Alabama

Despite the recent success of Alabama shrimp farmers in culturing the Pacific white shrimp, Litopenaeus vannamei , in inland low-salinity waters there is large variability in growth and survival among ponds. Farmers suspect that high mortality occurs during the first weeks of culture following stocking of postlarvae (PL). In order to determine the effect of pond ionic composition on PL growth and survival, three trials were carried out at a shrimp farm. Trials 1 and 2 evaluated PL growth and survival over 21- and 28-day periods, respectively, using static water from different production ponds. Trial 3 evaluated growth and survival over 7, 14, 21 and 28 days using water from one production pond. Results suggest that initial mortality (8–10%) is attributed to the acclimation process and occurs immediately following stocking. Pond-to-pond variations in ionic profiles could be a contributing factor but are not likely the major reason for variable 'survival'. Mortalities after stocking appear to occur quickly as there were no differences in survival from 7 to 28 days post stock. Variable survival is likely due to a combination of reasons including environmental factors, but is largely due to poor handling of PL and stocking errors that produce perceived reductions in survival.