Genetic parameters for spawning and growth traits in the Pacific white shrimp (Penaeus (Litopenaeus) vannamei)

The aim of this study was to estimate genetic parameters for spawning traits and growth traits in a breeding line of Pacific white shrimp, Penaeus (Litopenaeus) vannamei, selected for growth and survival. Traits studied were number of eggs (NE) and number of nauplii (NN) and female body weight at insemination (FWI) and body weight at 130 days of age (BW130). Genetic parameters were estimated using a multivariate animal model. Heritability for NE and NN were estimated as 0.13 ± 0.04 and 0.03 ± 0.04 respectively. The contribution to NN total variation due to ‘factors associated with male’ effect was estimated as 0.47 ± 0.07. In the cases of FWI and BW130, heritability was estimated as 0.44 ± 0.08 and 0.19 ± 0.03 respectively. Genetic correlation between FWI and NE was estimated as 0.49 ± 0.15, between FWI and NN as 0.54 ± 0.39 and between NE and NN as 0.27 ± 0.41, whereas the genetic correlations of FWI, NE and NN with BW130 were 0.30 ± 0.13, ?0.21 ± 0.19 and ?0.25 ± 0.38 respectively. Although it is important to perform more studies on this issue, our results found no evidence of a genetic antagonistic effect between female reproductive traits and body weight at harvesting (130 days of age) in P. vannamei.     

凡纳滨对虾不同世代生长性状的变异

对从美国进口的选育凡纳滨对虾(Litopenaeus vannamei)海南群体(进口亲虾繁育的第1世代,G1)、山东和饶平群体(G2)、湛江2和湛江3群体(G3)、湛江1和上海群体(G4)共7个养殖群体4个世代1150个个体的生长性状体长和体重进行了分析。7个群体的平均体长(范围)分别为14.76(13.25~15.99)、8.46(6.28~10.48)、9.24(4.28~10.70)、7.75(5.13~9.36)、11.38(8.13~14.12)、5.25(3.47~6.83)和7.14(4.14~9.00),变异系数分别为0.04、0.08、0.08、0.09、0.12、0.14、0.14,平均体重(范围)分别为33.41(24.33~39.74)、5.19(1.80~9.68)、6.95(3.18~11.34)、4.62(1.52~9.87)、15.03(6.00~26.96)、1.47(0.48~3.42)、3.29(0.49~6.20),变异系数分别为0.10、0.23、0.21、0.27、0.32、0.39、0.36。体长和体重的变异系数随着繁育世代的增加而增加,其中体重的变异系数每繁殖1代增加10%,其第1代的变异系数与美国选育的亲本群体相同。体长、体重相关与回归分析表明,体长与体重相关极显著(P<0.01),体长和体重的回归方程为W=0.01L2.93。表明随着繁育世代的增加,生长性状逐代分化。     

Genetic parameters and response to selection for harvest body weight of pacific white shrimp,Litopenaeus vannamei

Genetic parameters and response to selection were estimated for the harvest body weight of Litopenaeus vannamei. The data consisted of 24 072 progeny from 178 sires and 171 dams in two generations (G₀ and G₁) with a nested mating structure. All families were randomly divided into two groups and then cultured in two farms at different locations (Huanghua and Qingdao). The heritability estimates from G₀ and G₁ were 0.278 ± 0.136 and 0.423 ± 0.065 respectively. Over two generations, the heritability estimate was 0.335 ± 0.087, and the common environmental effect was 0.084 ± 0.031. A bivariate animal model was used to estimate variance and covariance components, whereby the body weight in the two farms was treated as a genetically distinct trait. Genetic correlation was close to unity (0.943 ± 0.066), indicating that a genotype by environmental interaction for harvest body weight was small. The response to selection in harvest body weight was estimated using two methods (the realized and predicted responses). The realized response was estimated from the difference in the least squares means of body weight for the selection and control populations. The predicted response was obtained from the difference in the mean estimated breeding values between generations. The realized response was 2.30%, while the predicted responses were 2.00% and 1.37% for within‐ and across‐generation datasets using two sets of genetic parameters respectively. The results would provide crucial information in pacific white shrimp breeding programs in China.     

Genetic differentiation in hatchery strains and wild white shrimp Penaeus (Litopenaeus) vannamei (Boone, 1931) from northwest Mexico

Ten enzymatic systems were analyzed to determine allozyme genetic differentiation among three hatchery strains (A, B and C) of white shrimp, Penaeus (Litopenaeus) vannamei, commonly used in shrimp farming in northwest Mexico. A wild population from northern Sinaloa was used as a reference. Fifteen loci were detected, nine of which were polymorphic (ACP-1*, ACP-2*, AKP-2*, EST-2*, EST-3*, EST-4*, EST-5*, LAP*, and LDH*). Polymorphism of A, B and C were 53, 53, and 40%. The mean observed heterozygosity per locus was 0.071, 0.093, and 0.050 without any significant difference among them or with respect to the wild population (0.056). In all samples observed heteroxygosity was smaller than expected. The mean number of alleles per locus was 1.8 in A and 1.87 in both B and C. Strain A was the only sample without rare alleles. Only EST-3* and LAP* of strain A were in Hardy-Weinberg equilibrium; the other loci were in disequilibrium in all samples. High inbreeding values and heterozygote deficiencies were detected in all samples. Distribution of allelic frequencies was heterogeneous among the samples (G-test), involving all polymorphic loci and suggesting a genetic differentiation. According to Fst, a moderate genetic differentiation (7.4%) was detected among the samples. Greater differences were between strains A and C. Based on genetic distance, the samples were grouped into two pairs, B–C and A-wild. Strain A is a young strain related to the wild sample, whereas strains B and C have a different geographic origin than the wild sample.     

Genetic analysis of the Pacific white shrimp (Litopenaeus vannamei): heterosis and heritability for harvest body weight

The aim of this study was to estimate heterosis and heritability for harvest body weight of the Pacific white shrimp (Litopenaeus vannamei) measured at commercial farm conditions. Heterosis and heritability were estimated using a base population from diallel crosses of eight introduced strains. The base population included 9936 shrimp from 207 families that were produced with 188 sires and 172 dams using a nested mating design by artificial insemination. Heterosis was calculated basing on the least squares means (LSM) of harvest body weight. The results showed that most of the hybrids (75%) have positive heterosis for harvest body weight, which ranged from ?13.36% (UA2 × UA5) to 13.80% (UA6 × UA5) with a mean of 2.41%. The high amount of heterosis manifested in the hybrids indicated the usefulness of these hybrids for improving the growth. Variance components and heritability for harvest body weight were estimated using an animal model. The heritability estimate for harvest body weight was 0.092 ± 0.082 (h²) when genetic groups were excluded from the pedigree, but it was decreased when genetic groups were included in the pedigree ( = 0.066 ± 0.050), implying that there are strain additive genetic effect and heterosis in the base population. However, the heritability estimates for harvest body weight were significantly different from zero (P < 0.05) and there was no significant difference between h² and (P > 0.05). The results from this study indicated that significant improvement for growth is possible through cross‐breeding and selective breeding in L. vannamei.     

Quantitative genetic parameter estimates for size and growth rate traits in Pacific white shrimp, Penaeus vannamei (Boone 1931) when reared indoors

Heritabilities of size traits and growth rate traits, as well as genetic, phenotypic and environmental correlations were estimated at three ages for a captive population of Pacific white shrimp (Penaeus vannamei) grown indoors. A covariate, mean size or mean growth rate during early growth in individual tanks before tagging and mixing, was introduced in the analyses to reduce the error caused by unique previous growth conditions on variance components. Heritabilities of size traits increased with age, with the h² for TL, AL, TWt and Wi1AS being 0.15, 0.20, 0.20 and 0.22, respectively, at 17 weeks, increasing to 0.28, 0.33, 0.34 and 0.35 at 29 weeks of age. Heritabilities of growth rate traits estimated between consecutive growth periods decreased from the first (h² for ΔTL 0.65, ΔAL 0.71, ΔTWt 0.63 and ΔWi1AS 0.84) to the second period (h² for ΔTL 0.34, ΔAL 0.50, ΔTWt 0.54 and ΔWi1AS 0.52). Phenotypic correlations were always larger than genetic correlations for both, size and growth rate traits. Genetic correlations between size traits within age were high (r_G >0.95), but those between the same size trait at different ages decreased as the age difference increased in spite of a consistently high environmental correlation (r_E 0.80–0.85) between the same trait at different ages. Phenotypic and genetic correlation's between the same growth rate trait at the two different growth periods evaluated were negative or zero (r_G TL –0.26, AL –0.24, Wi1AS 0.00) with the exception of total weight (r_G TW 0.35) and the environmental correlations between growth periods were also low (r_E 0.13–0.32).     

Proteome modifications of Pacific white shrimp (Litopenaeus vannamei) muscle under biofloc system

The objective of the study was to examine the effects of biofloc technology on the muscle proteome of Litopenaeus vannamei. Two biofloc treatments and one control were compared: biofloc‐based tanks under zero‐water exchange fed with 150 g/kg crude protein (BF15), or with 250 g/kg crude protein (BF25) diets, and clear water tanks with 50% of daily water exchange stocked with shrimp fed with similar amount of a 250 g/kg crude protein diet, referred to as control. The shrimp (5.28 ± 0.42 g) were divided into the 300‐L fibreglass tanks (water volume of 200 L) at a density of 35 shrimp per tank and were cultured for 35 days. The biofloc groups displayed better growth and survival compared to the control. The muscle tissue from the control and BF25 groups was subjected to proteomic analysis. Lactate dehydrogenase, enolase, arginine kinase, mitochondrial ATP synthase subunit alpha, mitochondrial ATPase inhibitor factor 1 precursor, serpin 3 and myeloid differentiation factor 88 had an increased abundance in the BF25 group, while myosin heavy chain type 1 and myosin heavy chain type 2 showed a decreased abundance. The results indicate that biofloc technology could alter the expression of proteins involved in structure, metabolism and immune status of cultured shrimp.     

Genetic variation of white spot syndrome virus (WSSV) in Pacific white shrimp Litopenaeus vannamei (Boone 1931) culture of Thailand

White spot syndrome virus (WSSV) is highly pathogenic to penaeid shrimp and has caused significant economic losses in the shrimp farming industry in Thailand. Genotyping analysis was done in 124 WSSV isolates from cultured Pacific white shrimp Litopenaeus vannamei. These samples were obtained during 2007–2014 from eight provinces in Thailand. We investigated five variable loci in the virus genome: deletions in two variable regions, VR14/15 and VR23/24, and three variable number tandem repeats (VNTR) located in open reading frame (ORF) 75, 125 and 94. WSSV genotype was characterized as (X_14/15, X_23/24) (N₇₅‐N₁₂₅‐N₉₄) where X is the number of base pair deletion in the variable region and N is the number of repeat units (RUs) in a specific ORF. The deletion pattern in VR14/15 and VR23/24 regions characterized three WSSV genotypes. The most prevalent genotype was (5950_14/15, 10971_23/24), and it was found in all studied areas. At least 33 genotypes of WSSV were analysed based on 3 VNTR loci, indicating that the VNTRs of WSSV genome are highly variable. From 124 WSSV samples, two samples presented the characteristic of all five variable loci similar to WSSV collected during 2010 in Saudi Arabia (5950_14/15, 10971_23/24) (3₇₅‐6₁₂₅‐7₉₄). Many different WSSV genotypes shown in this study as compared to previously reported genotypes in Thailand suggests current status of disease epidemiology, as well as probable movements of WSSV between countries.     

零换水工厂化养殖模式下凡纳滨对虾生长和存活性状遗传参数估计

针对凡纳滨对虾生长和存活性状,本实验估算了零换水养殖模式下的遗传参数,分析该模式与大换水量养殖模式的基因型与环境互作效应(genotype by environment interaction effect,G×E),为后续留种、配种方案制定等育种规划工作提供依据。通过人工授精技术,定向交尾建立了51个凡纳滨对虾全同胞家系。利用荧光标记识别家系,采用零换水养殖模式混养52 d后,测量并记录2个养殖池中的3822尾存活虾的体质量、体长及性别等信息。采用线性(广义线性)混合效应模型和REML算法,基于个体动物模型和父母本阈值模型估计生长和存活性状的方差组分和遗传力。结果显示,收获体质量和存活率具有较大变异系数。收获体质量和体长的遗传力分别为0.49±0.08和0.43±0.07,均属于中高遗传力(h2≥0.15);存活的遗传力为0.11±0.03,属于低遗传力(h20.15)。收获体质量与收获体长的遗传相关系数为0.98±0.01;收获体质量和体长与存活的遗传相关系数分别为0.31±0.15和0.34±0.15。对于体质量和存活性状,零换水和大换水量养殖模式间的遗传相关系数分别为0.62±0.11和0.65±0.11,G×E效应显著(K0.5)。综上所述,凡纳滨对虾在零换水养殖模式下存在较高的遗传变异,但是与大换水量常规养殖模式相比,家系间存在较大的重排序效应,因此针对该养殖模式应单独建立选育系进行新品种培育。     

Evaluation of Schizochytrium meal in microdiets of Pacific white shrimp (Litopenaeus vannamei) larvae

A feeding trial was conducted to assess the effects of dietary Schizochytrium meal supplementation on survival, growth performance, activities of digestive enzymes and fatty acid composition in Pacific white shrimp (Litopenaeus vannamei) larvae (initial body weight 4.21 ± 0.10 mg). Four isonitrogenous and isolipidic diets were formulated to contain graded levels of Schizochytrium meal: 0% (S0, the control diet), 2% (S2), 4% (S4) and 6% dry matter (S6). Results showed that there was no significant difference in survival of shrimps among dietary treatments (P > 0.05). Shrimps fed diets with 2% and 4% microalgae meal had significantly higher specific growth rate (SGR) than that of shrimps fed diets with 0% and 6% microalgae meal, and no significant differences were observed between shrimps fed diets with 2% and 4% microalgae meal (P > 0.05). Activity of trypsin in the pancreatic and intestinal segments, and activity of amylase in the pancreatic segments were not significantly affected by dietary microalgae meal levels (P > 0.05). Specific activities of both alkaline phosphatase and leucine‐aminopeptidase in intestine and purified brush border membrane of intestine were significantly higher in shrimps fed diet with 2% microalgae meal (P < 0.05). There were no significant differences in C18:2n‐6, n‐3 fatty acids, n‐6fatty acids, PUFA and n‐3/n‐6 in muscle samples among dietary treatments. C16:1n‐7, C18:1n‐9, MUFA, C18:3n‐3 and C20:5n‐3 decreased, however, C20:4n‐6 increased in the muscle as dietary microalgae meal level increased. In conclusion, 4% Schizochytrium meal in microdiets of shrimps can improve growth performance and may be a valuable additive in the microdiets of shrimps.     

Genetics of the resistance to hypoxia in postlarvae and juveniles of the Pacific white shrimp Penaeus (Litopenaeus) vannamei (Boone 1931)

The genetics of resistance to hypoxia was studied in shrimp using two approaches, heritability estimation and heterosis estimation. The heritability of resistance to hypoxia was evaluated in two populations of shrimp postlarvae families produced at different times, and heterosis was estimated through the reciprocal crosses of two genetically distinct shrimp populations at postlarvae and juvenile. Heritabilities of resistance to hypoxia in postlarvae were large and out of the range for the heritability (1.73±0.09, 1.35±0.14) regardless of population evaluated. When covariates (survival and mean total weight to postlarvae 15 days old) were introduced in the analyses to correct for effects during rearing of the larvae, the heritabilities did not change substantially for the first population, but were decreased for the second population depending on the covariate used for correction (1.15±0.10 and 1.08±0.11 for the covariates survival and total weight respectively). Those large heritabilities could be explained by the presence of confounded common environmental variance in the estimated genetic variance (maternal effects), or by dominance and epistatic effects being an important part of the estimated genetic variance. Heterosis analyses dissecting maternal effect indicated that both effects, maternal and dominance, are involved in the resistance to hypoxia at postlarvae, and that although maternal effects decreased at juvenile stage, the trait was still characterized by presenting a large heterosis value (62–65% for postlarvae and 36% for juveniles).     

凡纳滨对虾生长性状遗传参数的估计

采用平衡巢式设计方法和人工授精技术,每个雄体配3个雌体,构建了18个父系半同胞家系和54个母系全同胞家系.分别测定了每个母系生长到5月龄60个全同胞个体的体质量、全长、体长、头胸甲长、头胸甲宽、头胸甲高、第一腹节背高、第三腹节背高和第一腹节背宽9个生长性状,应用数量遗传学原理,采用方差和协方差分析的方法,估算了5月龄凡纳滨对虾生长性状的遗传力及各性状间的遗传相关及表型相关.结果表明,利用父系半同胞组内相关法估计的遗传力是凡纳滨对虾各生长性状遗传力的无偏估计值.其中,体质量、全长、体长、头胸甲长、头胸甲宽、头胸甲高、第一腹节背高、第三腹节背高和第一腹节背宽狭义遗传力的估计值分别为0.460、0.392、0.303、0.234、0.251、0.330、0.282、0.321和0.356,属于中高等遗传力范围,显示出较高的选择育种潜力.基于父系半同胞遗传协方差组分及表型协方差分别估计的各性状间的遗传相关及表型相关表明,各个性状间均表现出高的正相关,其中遗传相关在0.750 ～0.976,体质量—全长的遗传相关为最大(0.976),全长—第三腹节背高的遗传相关为最小(0.750),表型相关为0.507～0.947,体质量—全长为最大(0.947),第一腹节背高—头胸甲高为最小(0.507).经t检验,各性状间遗传相关及表型相关均达到极显著水平,表明以任意一个生长性状为参数进行选育,均可达到改良凡纳滨对虾生长情况的效果.     

Response of Pacific white leg shrimp (Litopenaeus vannamei) on exposure to aflatoxin in feed

A research was accomplished to find the response of Litopenaeus vannamei to different levels of aflatoxin in feed. Two hundred live L. vannamei were collected from a shrimp breeding station located on the south of Tiab, in Hormozgan province. Aflatoxin contaminated diets with 0.18, 96.21, 184.74, 711.35, 977.11 and 1605.61 µg/kg were prepared and shrimps in six experimental groups with three replicates in each group for 28 days were fed with these diets. The results showed that activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) in haemolymph and histological alternation index (HAI) value of hepatopancreas in treatement groups were significantly higher than control group (p < 0.05). Decreased total protein and fat in haemolymph and increased protein in muscle of treatment shrimps were observed compared with the control group (p < 0.05). The HAI value of 977.11 and 1605.61 μg/kg groups were significantly higher than the control and other groups (p < 0.05). Histological study results showed that calculated HAI (more than 100) in these groups as a result of necrosis lesion observed in hepatopancreas. Moderate to severe alteration was recognized in group fed by 711.35 µg/kg aflatoxin. Minor aflatoxin levels (0.18, 96.21 and 184.74 µg/kg) caused inappreciable tissue alteration. This study demonstrated body composition, haemolymph parameters and histology of hepatopancreas were affected in the aflatoxin‐contaminated feed when the concentration was over 955.14 µg/kg.     

Hydrolysed salmon meal as a replacement for salmon meal in practical diets for Pacific white shrimp (Litopenaeus vannamei)

A series of growth, feed stability and consumption trials were conducted to evaluate the efficacy of salmon by‐product in practical diets for Pacific white shrimp, Litopenaeus vannamei. This included a salmon by‐product meal (Salmon meal: SM) and a silage hydrolysate (hydrolysed salmon meal: HSM). The basal diet containing 120 g/kg SM was incrementally replaced (0. 25, 50, 75, 100%) by HSM to produce five test diets used in two trials. A sixth diet was included which evaluated gelatin supplementation (trial 1) or pH neutralization (trial 2). In trial 1, each diet was produced using two processing conditions (laboratory extruded and formed with meat grinder) and offered to shrimp in a clear water system. The results demonstrate that up to 50% of the SM can be used to replace with HSM; however, further increases resulted in reduced performance of shrimp. The addition of gelatin reduced leaching but there was limited effect of processing on leaching. There were no detectible effects of pH adjustment of the diets. Results indicated that the growth performance of shrimp has not influenced by HSM up to 60 g/kg to replace 50% of the SM in practical diets; however, higher levels resulted in significant decrease in performance.     

Utilization of fermented soybean meal for fishmeal substitution in diets of Pacific white shrimp (Litopenaeus vannamei)

Two feeding experiments were carried out to evaluate the utilization of fermented soybean meal (FSBM) in diets for Pacific white shrimp (Litopenaeus vannamei). The digestibility experiment was assessed with four diets: a reference diet and three ingredient test diets containing FSBM, local and Chilean fishmeal. The growth experiment was conducted including four isonitrogenous and isocaloric diets formulated to contain the graded levels of dietary fishmeal (0 g/kg, 150 g/kg, 300 g/kg and 450 g/kg) substituted by FSBM. The results indicated that ADCs of crude protein in FSBM were high (888.4 g/kg) and equal in local fishmeal but little lower than Chilean fishmeal while without any significant differences observed in ADCs of crude lipid among the different test ingredients. After 75 days of growth trial, survival rates and feed conversion ratio were not significantly different among the experimental treatments. However, shrimp fed the diets with increasing inclusions of FSBM had a tendency to reduce weight gain and specific growth rate. Based on the correlation between weight gain and substituted fishmeal level analysed by broken‐line regression, the optimum level of fishmeal replaced by FSBM in diet was 253.6 g/kg without adverse effects on growth and feed utilization of Pacific white shrimp.     

Effects of the probiotic Bacillus subtilis (BP11 and BS11) on the growth and survival of Pacific white shrimp,Litopenaeus vannamei

The effect of Bacillus S11 (BS11)‐ and/or Bacillus P11 (BP11)‐supplemented feeds on the growth performance, survival, immunoenhancement and disease resistance of cultured Pacific white shrimp, Litopenaeus vannamei, was evaluated. Four feeding treatments of (i) regular feed (control), (ii) BS11‐supplemented feed, (iii) BP11‐supplemented feed and (iv) BS11‐and BP11‐supplemented feed were prepared and used for shrimp cultivation in closed recirculating cement tanks (~400 L) in two trials, one for juvenile and PL‐30 shrimp at 60 and 90 days, respectively. The results showed that BS11 gave a higher probiotic potential than BP11 for both age groups of L. vannamei in cultivation, because the average weight and survival of shrimp fed BS11‐supplemented feed were significantly higher (P < 0.05) than those of the control and the other two groups. The survival of shrimp fed either BS11‐or both BS11‐and BP11‐supplemented feed was significantly higher (P < 0.05) than that of the control group. In addition, the highest total haemocyte and granular haemocyte counts and phenoloxidase activity were found in shrimp fed with the BS11‐supplemented feed. After challenge with Vibrio harveyi 639 (~10⁷ CFU mL^?1) by immersion, the lowest cumulative death (%) and disease resistance were clearly found in shrimp fed with the BS11‐supplemented feed.     

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.     

氨氮胁迫下白斑综合征病毒对凡纳滨对虾的致病性

为了评价养殖水环境中氨氮(NH_4-N)对凡纳滨对虾(Litopenaeus vannamei)的危害性,开展了NH_4-N胁迫对凡纳滨对虾感染白斑综合征病毒(WSSV)后的死亡率、WSSV增殖速率和对虾主要免疫相关酶活性影响的实验。在NH_4-N胁迫质量浓度为15.6 mg·L-1,分别注射2×105和2×106个WSSV粒子,结果显示,NH_4-N胁迫下注射2×105个WSSV粒子的凡纳滨对虾第144小时死亡率达到53.3%,显著高于无胁迫组(40.0%)。对虾鳃组织WSSV荧光定量PCR检测结果显示,NH_4-N胁迫下凡纳滨对虾鳃组织内WSSV的增殖加快。此外,免疫相关酶活性结果显示,NH_4-N浓度突变会促使对虾血清中酚氧化酶(PO)、酸性磷酸酶(ACP)和碱性磷酸酶(AKP)活性短暂升高后持续降低。由此可见,NH_4-N胁迫会加快WSSV在患病凡纳滨对虾体内的增殖,导致更高死亡率,这可能是因为胁迫造成了对虾免疫相关酶活性降低和抗病原感染能力下降。