A reference diet for nutritional studies of the giant tiger prawn Penaeus monodon

Critical evaluation of the nutrient requirements of an animal requires the use of a purified reference diet. In this study, the effects of either a purified reference experimental diet, a practical experimental diet and an imported commercial diet on the growth and nutritional condition of the prawn Penaeusmonodon were compared. The reference diet, based on widely available ingredients of consistent and known nutrient content, supported excellent prawn growth when fed either ad libitum or subsatiation (≈ 75% of the mean initial satiation intake). In a 6-week experiment, growth (as percentage increase in weight, ± SEM) of the prawns fed ad libitum differed significantly (P < 0.05) between each diet in the order: practical (142 ± 10%), reference (121 ± 9%) and commercial (91 ± 16%). Growth for the reference diet fed subsatiation was 116 ± 4%, and was not significantly (P > 0.05) different from that obtained with the same diet fed ad libitum. Dry matter food conversion for the reference diet was significantly (P < 0.05) better when fed subsatiation (1.58 ± 0.08) rather than ad libitum (2.08 ± 0.06) and better than that obtained with either the practical (3.40 ± 0.15) or commercial (3.02 ± 0.28) diets, which were not significantly (P > 0.05) different from each other. The lipid content (mg g^–1 of prawn) of the digestive gland of prawns fed ad libitum was similar for reference, practical and commercial diets (6.03 ± 0.38, 4.92 ± 0.90 and 4.92 ± 0.95 mg g^–1 of prawn, respectively), but significantly (P < 0.05) higher than that for the reference diet fed subsatiation (4.73 ± 0.38 mg g^–1 of prawn).     

Effects of adding sucrose on Penaeus monodon (Fabricius, 1798) growth performance and water quality in a biofloc system

A 60‐day indoor growth trial was conducted to study the effects of biofloc on the growth performance of a Penaeus monodon (Fabricius, 1798), water quality and biological indicators including biofloc volume, chlorophyll‐a, heterotrophic bacteria and Bacillus quantity. Two concentrations of sucrose (0 and 75%) were added daily to the P. monodon culture systems (2.94 ± 0.11 g), which were conducted indoors in fibre‐glass tanks (500 L). Results showed that the final body weight and weight gain of the adding 75% sucrose group were significantly higher (P < 0.05) than that of the control, as well as significantly (P < 0.05) improved specific growth rates and survival rates, and reduced feed coefficient. Adding 75% sucrose promoted heterotrophic bacteria, Bacillus and phytoplankton reproduction, and significantly (P < 0.05) reduced the concentration of ammonia‐N (NH₄‐N), nitrite‐N (NO₂‐N) and nitrate‐N (NO₃‐N). The changes of water quality indicators in the two groups showed the similar trend at the end of the experiment, and the ammonia‐N, nitrite‐N, nitrate‐N and phosphate‐P concentrations in the 75% sucrose group were significantly (P < 0.05) lower than those of the control group, Chlorophyll‐a concentrations peaked at 389.12 μg/L in the biofloc sucrose group at 18:00 h, and heterotrophic bacteria peaked 8 h after sucrose was added. The addition of sucrose also reduced the pH of the water. Our research showed that adding sucrose promoted biofloc formation and shortened the formation time; increased the number of heterotrophic bacteria and algae which might play a role in improving water quality by assimilating ammonia‐N and other harmful substances in the water; supplemented food for P monodon growth; and reduced the feed coefficient.     

The use of passive acoustics to measure feed consumption by Penaeus monodon (giant tiger prawn) in cultured systems

The feeding inefficiencies associated with intensively cultured prawn systems have a significant financial cost and environmental impact. Initial trials of a commercial system using sound to manage feeding within cultured systems have achieved promising results with an impressive food conversion (food weight/biomass) ratio of 1.42. Whilst these results demonstrate the potential benefit of employing passive acoustics for feed management, the underlying technologies are not well understood by industry or the research community. Consequently, a sound based study of feeding tiger prawns is conducted to investigate the key challenges associated with passive acoustic approaches; sound detection and feed demand estimation. The study finds that tiger prawns produce impulsive sound signatures during feeding that can be used as a proxy of feeding activity. Spectral features of the feeding signatures can be used to detect feeding activity within acoustically complex farm ponds (feeding signal to noise ratios less than −40 dB), given they are largely disjoint from the background noise spectrum (aerators) across all farm recordings. One of the potential challenges identified with sound based feed detection is that other sources of event driven interference arise (i.e. rain and faulty aerators), which can be misclassified as feeding. Whilst our investigation indicates that sound based detection of feed events are plausible, it is unclear how accurate it is to estimate the quantity of consumed pellets from feeding sound. Our study provides evidence to support its feasibility, given the temporal evolution of feeding sounds and pellet consumption were linearly related across tank and pond based feeds, respectively (R² = 0.95 and R² = 0.96).     

Effects of biofloc under different carbon sources and protein levels on water quality,growth performance and immune responses in black tiger shrimp Penaeus monodon (Fabricius, 1978)

A 75‐day 2 × 3 factorial experiment was conducted to evaluate the effect of two levels of dietary protein (32 and 40%) and two different carbon sources (rice flour–R and molasses‐M), and without carbohydrate (control–C) in black tiger shrimp Penaeus monodon juveniles (3.37 ± 0.04 g) at 100 nos. m^?3 in 100 L fibre reinforced plastic tanks. Biofloc volume and total suspended solid were higher in molasses added groups (32 + M and 40 + M) followed by rice flour (32 + R and 40 + R) and controls (32C and 40C). Molasses and rice flour addition significantly reduced (P < 0.01) the total ammonia–N compared to controls. The highest Vibrio, Bacillus and Lactobacillus counts were recorded in 40 + M, 32 + M and 32 + R respectively. Among the treatments, significantly higher (P < 0.01) final body weight was recorded in 40 + R (8.5 ± 0.3 g), 40 + M (7.8 ± 0.3 g) and 32 + R (7.5 ± 0.4 g) compared to control groups, 32C (6.1 ± 0.3 g), 40C (6.4 ± 0.3 g) and molasses added group, 32 + M (5.7 ± 0.4 g). Rice flour supplementation significantly increased (P < 0.01) the total haemocyte count (×10⁶ cells mL^?1) in 32 + R (45.7 ± 3.7) and 40 + R (44.3 ± 3.1) compared to controls, 32C (27.3 ± 3.4) and 40C (25.8 ± 0.9). Similarly, higher superoxide dismutase, catalase, serum protein and glucose were recorded in the rice flour added groups, 40 + R followed by 32 + R. Among the treatments, the highest level of prophenoloxidase (OD 490 nm, 0.3 ± 0.0) and survival after challenge with Vibrio harveyi (55.6%) was observed in 32 + R. The study elucidates that rice flour addition produces optimum level of biofloc with better growth and immune responses compared to molasses and control. Furthermore, rice flour addition at 32% protein level could replace 40% protein feed.     

Effects of dietary mannan oligosaccharide on growth,survival, physiological,immunological and gut morphological conditions of black tiger prawn (Penaeus monodon Fabricius 1798)

The aim of this experiment was to determine the effects of dietary inclusion with mannan oligosaccharide (Bio‐Mos, Alltech, Nicholasville, KY, USA) on growth, survival, physiological and immunological conditions and gut morphology of the black tiger prawn (Penaeus monodon). Five diets supplemented with MOS at 0 g kg^?1 (control diet), 1, 2, 4 and 8 g kg^?1 were fed to the prawn juveniles (0.4 ± 0.06 g, total weight) for the duration of 63 days. Growth was the highest (P < 0.05) when the prawns were fed the 1 g kg^?1 MOS included diet. Wet tail muscle index (Tw/B), dry tail muscle index (Td/B) and tail muscle protein (Tmp) were higher (P < 0.05) in the prawns fed MOS included diets MOS compared with the prawns fed the control diet. Total haemocyte counts (THCs) of the prawns fed MOS included diets were higher (P < 0.05) than THCs of the prawns fed the control diet. Epithelium layer and epidermal cell density of the gut of the prawns fed 1 g kg^?1 and 2 g kg^?1 MOS diets were better than the prawns fed the control and other MOS diets. The results imply a positive effect of dietary supplementation of 1–2 g kg^?1 MOS in the culture of black tiger prawns.     

Integration of substrate in biofloc based system: Effects on growth performance,water quality and immune responses in black tiger shrimp,Penaeus monodon culture

To evaluate effect of substrate integration in biofloc based system, a 52‐day growth experiment was conducted using black tiger shrimp, Penaeus monodon juveniles (3.32 ± 0.07 g). The factorial design consisted of floc, F (with or without) as first factor and substrate (bamboo mat, B; nylon mesh, N; and without substrate) as second factor. This resulted six treatments; F + B, F + N, F, B, N and a control without biofloc and substrate. Shrimps were stocked at 110 nos. m^–3 in Fibre Reinforced Plastic (FRP) tanks and, rice flour was used as carbon source in biofloc based treatments. Incorporation of nylon mesh and bamboo mat in biofloc system trapped 31.3%–38.6% and 8.5%–13.5% total suspended solids respectively and reduced bottom solid deposition. Among the substrate based groups, significantly better development of biofilm with higher microbial population noticed in F + B compared with nylon mesh. Similarly, significantly higher final growth (p < 0.01) was recorded in F + B system followed by F + N while no significant difference in body weight recorded among floc, F or substrate based groups (B, N). Biofloc and substrate integration (F + B and F + N) resulted significantly (p < 0.01) lower feed conversion ratio compared to control and floc. Incoporation of bamboo substrate in biofloc, (F + B) improved shrimp immune responses through higher hemocyte counts and prophenoloxidase activity compared to other treatments. The study revealed that integration of substrate in the biofloc system improved growth performance, FCR and immune parameters in shrimp by trapping the suspended biofloc particles, better water quality parameters, enhanced biofilm growth and provision of quality natural food.     

饲料中不同蛋白含量对斑节对虾幼虾生长及消化酶的影响

选用鱼粉、豆粕和大豆浓缩蛋白为主要蛋白源,配置6个蛋白质水平梯度(36%、38%、40%、42%、44%、46%)的饲料,对斑节对虾幼虾(1.03±0.02 g)进行56 d养殖实验,研究饲料中不同蛋白含量对斑节对虾幼虾生长及消化酶活性的影响。结果表明,(1)摄食40%蛋白饲料的斑节对虾的增重率(417.35%)和特定生长率(2.93%·d-1)获得最大值并显著高于36%蛋白饲料组(P<0.05),但与其它各组无显著差异。饲料蛋白水平在38%时的饲料系数(1.88)最低且与44%和46%蛋白饲料组存在显著性差异(P<0.05)。(2)虾体蛋白含量在44%饲料蛋白组最高,且显著高于36%、38%和42%组(P<0.05)。(3)斑节对虾的蛋白质表观消化率在42%蛋白组最高,44%和46%蛋白饲料组的蛋白质效率显著低于36%～42%蛋白饲料组(P<0.05)。(4)随着蛋白水平的提高,斑节对虾的蛋白酶活性呈现由高到低波动的趋势;淀粉酶活性随着饲料蛋白含量的升高而呈下降趋势,胃淀粉酶活性在38%～42%蛋白饲料组显著高于36%蛋白饲料组(P<0.05),肠淀粉酶活性在42%蛋白饲料组显著高于其它各组(P<0.05)。通过饲料中蛋白质含量与增重率的回归分析,斑节对虾幼虾饲料蛋白的适宜含量为39.70%。     

Genetic evidence for a single stock of giant tiger prawns Penaeus monodon in demarcated prawn fishing zones of Tanzania

Tanzania demarcated three prawn fishing zones along its coast to spread fishing pressure and help the prawn fishery to recover. However, it is unknown whether the demarcated zones correspond to the genetic stock structure of the world's most commercially important prawn, the giant tiger prawn (Penaeus monodon). Thus, this study used partial mitochondrial control region sequences (534 base pairs) to test the hypotheses that (1) giant tiger prawns in Tanzania's demarcated prawn fishing zones are a single stock and (2) the giant tiger prawns on the Tanzanian coast experienced a recent demographic expansion. The sequences showed high haplotype diversity (h = 0.998–1.0) and low nucleotide diversity (θ_π = 1.89%–2.24%). The neutrality test and mismatch analysis showed that the hypothesis of recent demographic expansion could not be rejected. The analysis of molecular variance revealed low and insignificant fixation indices between the zones (F_ST = 0.00025, p > 0.05; Φ_ST = −0.00027, p > 0.05), suggesting that the three demarcated fishing zones constitute a single stock and that fishers may be targeting the same stock. Furthermore, it was discovered that Zone 2 has the potential to replenish depleted areas; thus, it should be prioritised in future conservation planning.     

Lagenidium callinectes (Couch, 1942) infection and its control in cultured larval Indian tiger prawn, Penaeus monodon Fabricius

A mycosis in larval tiger prawn, Penaeus monodon Fabricius, is described for the first time from India. The hyphae of Lagenidium callinectes are contorted, irregularly branched, sparingly septate, and contain a cell wall and membrane, vacuoles, mito chondria, ribosomes, small and large vesicles, and Woronin bodies. The spores occur singly or in pairs. The fungal mycelium may either invade and embed itself in the tissues, or alternatively, replace all the muscle tissues of the infected larval P. monodon. Fungus infected, untreated populations of nauplii, zoea and myksis exhibited mortalities of 5.33 ± 0.55%, 24.68 ± 4.58% and 47.89 ± 0.27%, respectively. A 0.5 ppm treatment with trifluralin significantly reduced the mortality of infected larval populations (i.e. 1.1% nauplii, 3.28% zoea and 5.21% myksis mortality). Lagenidium sp. exhibited growth in potato dextrose agar medium and in Sabouraud's agar at 28 °C.     

Probiotic bacteria effects on growth and bacterial composition of black tiger shrimp (Penaeus monodon)

The effects of Bacillus probiotic forms on growth performance and microbiota were examined in black tiger shrimp (Penaeus monodon). Shrimp were fed for 120 days with one of three diets; a control diet alone or supplemented with either probiotics consisting of live‐sprayed Bacillus (LS) or freeze‐dried Bacillus (FD). Culturable bacteria counts in the hepatopancreas, intestine and culture water of the three treatments were relatively constant during the experiment. Bacillus numbers in the hepatopancreas and intestines of probiotic‐treated groups increased significantly (P < 0.05) after 30 treatment days and remained relatively constant afterwards while no appreciable changes occurred in the control. Shrimp supplemented with LS and FD contained significantly lower concentrations of Vibrio (P < 0.05) in the hepatopancreas, intestine and culture water, compared to those in the control group. Shrimp treated with LS and FD probiotic bacteria showed a significant improvement (P < 0.05) in specific growth rate (SGR) and feed efficiency (FE), compared to those of control group. Probiotic bacteria have the potential to enhance shrimp culture by reducing Vibrio and improving growth performance.     

Astaxanthin profiles and corresponding colour properties in Australian farmed black tiger prawn (Penaeus monodon) during frozen storage

The colour of commercial cooked black tiger prawns (Penaeus monodon) is a key quality requirement to ensure product is not rejected in wholesale markets. The colour, due to the carotenoid astaxanthin, can be impacted by frozen storage, but changes in colour or astaxanthin profile, during frozen storage, have not been studied in detail. Subsequently in this study, the aims were to define the astaxanthin (as cis, trans, mono‐ester and di‐ester forms) content, together with the colour properties, in both pleopods (legs) and abdominal segments. Changes in astaxanthin content and colour properties were further determined during frozen storage (?20°C). Total astaxanthin content was seen to decrease in all samples over time, with the rate of degradation being significantly greater (P < 0.05) in pleopods than abdomen. In both pleopods and abdomen, rate of degradation of esterified forms was significantly greater (P < 0.05) than non‐esterified forms. Hue angle (increase), a* value (decrease) and L value (increase) were all seen to significantly change (P < 0.05) during storage, with changes being more prevalent in the pleopods. The pleopods are the key indicator of astaxanthin and colour loss in cooked black tiger prawns and preservation strategies are required to preserve astaxanthin and colour during frozen storage.     

Effects of water exchange protocols on water quality,sedimentation rate and production performance of Penaeus monodon in earthen ponds

This study was carried out in farmers' fields to quantify the total water and consumptive water use in grow‐out culture of Penaeus monodon under recommended package of practice with two different water management protocols: T₁, with no water exchange and T₂, with regulated water exchange. Treatment‐wise estimated total water use, was 2.09 and 2.43 ha‐m 122 day^?1, while the computed consumptive water use index (m³ kg^?1 biomass) was 5.35 and 6.02 in T₁ and T₂ respectively. Lower rates of water exchange (T₂) showed significantly improved (P < 0.05) crop performance in terms of performance index (19.75 ± 0.75), production‐size index (74.1 ± 3.4), survival rate (80.13 ± 1.7%) and productivity (2.44 ± 0.08 t) over the zero water exchange. The shrimp pond water quality suitability index (WQSI) infers that regulated water exchange (T₂) improved the overall suitability of water quality for shrimp culture. WQSI up to 90 days of culture ranged between 7.5–9.0 in T₂, needs little management while in the last month of rearing, it was good with moderate management requirements. Treatment‐wise sediment load ranged between 50.4–56.3 m³ t^?1 shrimp biomass. High intensity of water exchange and low apparent feed conversion ratio influenced in lowering the sedimentation rate. Regulated water exchange protocol (T₂) performed well (higher net total water productivity and net consumptive water productivity) against no water exchange (T₁). A higher OV:CC ratio (ratio of the output value to the cost of cultivation) indicated that T₂ had a distinct edge over the T₁ protocol.     

Suitable dietary chitosan improves the growth performance,survival and immune function of tiger shrimp,Penaeus monodon

Different levels of dietary chitosan on growth performance, survival and stress tolerance to air exposure was studied in tiger shrimp, Penaeus monodon. Shrimp (mean initial wet weight about 1.16 g) were fed with six different diets (C0, C0.05, C0.1, C0.2, C0.3 and C0.4) containing six level of chitosan (0%, 0.05%, 0.1%, 0.2%, 0.3% and 0.4% respectively) in triplicate for 60 days. Growth performance [final body wet weight (FBW); weight gain (WG); biomass gain (BG)] of shrimp fed chitosan‐containing diets were higher (P < 0.05) than that of shrimp fed the basal diet, shrimp fed C0.1 diet showed the highest value of growth performance. Survival of shrimp in C0.1 and C0.2 diet groups were higher (P < 0.05) than that of shrimp in C0, C0.05 and C0.4 diet groups but without statistical difference (P > 0.05) in shrimp fed C0.3 diet group. Whole body and muscle lipid contents decreased with increasing dietary chitosan levels. Plasma total cholesterol and triglyceride contents of shrimp fed C0 diet was significantly higher (P < 0.05) than that of shrimp fed chitosan‐containing diets. Plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities of shrimp fed C0 diet were higher than those of shrimp fed chitosan‐containing diets. Digestive gland malondialdehyde (MDA) and carbonyl protein contents of shrimp fed chitosan‐containing diets were lower (P < 0.05) than that of shrimp fed C0 diet. Total haemocyte count of shrimp fed C0 diet was lower (P < 0.05) than that of shrimp fed chitosan‐containing diets. On the contrary, the haemolymph clotting time of shrimp fed C0 diet was higher (P < 0.05) than that of shrimp fed chitosan‐containing diets. In conclusion, all results suggested that dietary intake containing 0.1% and 0.2% chitosan enhanced the growth of shrimp, whereas a higher level than 0.3% and 0.4% decreased growth of shrimp. Second‐degree polynomial regression analysis of WG and BG indicated that the optimum supplement of dietary chitosan level should be 0.19–0.21%.     

A study of the arachidonic acid requirements of the giant tiger prawn, Penaues monodon

This study examined the dietary requirement of arachidonic acid (ARA) when that of linoleic acid (LOA), the natural precursor to ARA, was also satisfied with linolenic acid (LNA) and also with and without the other key dietary highly unsaturated fatty acids (HUFA). Growth by prawns fed diets supplemented with ARA was poorer than in diets where it was not present. Supplementation of ARA to diets with either optimized HUFA or just optimised poly unsaurated fatty acids (PUFA) (i.e. LOA, LNA) resulted in poorer growth. Growth was poorest by prawns (215 ± 13%) fed diets with ARA supplemented at 20% of the total fatty acids but including 7% LOA, 21% LNA and 4% of both eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Growth was best in prawns fed diets devoid of ARA but with 7% LOA and 21% LNA (350 ± 19%). Prawns fed the reference diet (348 ± 21%) and the other diet devoid of ARA but containing about 7% LOA, 21% LNA and 4% of both EPA and DHA (345 ± 18%) had similar growth. The growth responses were not effects of altered lipid or fatty acid digestibilities. Indeed supplementation of ARA to the diet marginally improved the digestibility of the total neutral lipid in the diet and the digestibilities of some other dietary fatty acids. The amount of lipid in the digestive glands of prawns fed with the diets was reduced by the inclusion of ARA in the dietary lipids. Composition of the lipids in the digestive gland (DG) of the prawns was almost directly related to the composition of their dietary lipids. The proportion of ARA in the total fatty acids increased with level of supplementation of dietary ARA. An increased level of dietary ARA reduced the proportion of EPA, DHA in the DG lipid and also the total n‐3 and n‐6 fatty acids in the DG lipid. The results of this study support that addition of ARA to the diet of Penaues monodon when the other key essential fatty acids (EFA) have been optimized, does not improve their growth performance. It is suggested that key cause for this response may lie in the importance of the balance of the n‐3 to n‐6 fatty acids in the diet of these animals.     

Nutrient efficacy of microalgae as aquafeed additives for the adult black tiger prawn,Penaeus monodon

This study used a small amount of several lyophilized microalgae (5% of dry weight in aquafeed) to feed adult black tiger prawns Penaeus monodon for 4 weeks, aiming to promote prawn health and nutritional level. Apart from slight increases in ash content, chlorophyll and β‐carotene in the microalgae‐added feeds, there was no difference from the commercial diet in their biochemical composition. After 28 days of feeding, prawn body length, body weight, condition index and also total amount of fatty acids and amino acids of the flesh (2nd abdominal segment) were not affected by microalgae additives as compared with the control commercial diet‐fed group (P > 0.05). Prawns fed the microalgae diets, however, had a relatively better survival rate than those fed the commercial control diet, although no statistical significance was detected in any except for the diatom Melosira sp. The elevated survival rate was concomitant with a higher phagocytosis rate and superoxide dismutase (SOD) activity in the immune response analysis, and significantly a higher astaxanthin content detected in the tail muscle of microalgae diet‐fed prawns (5.36–10.78 ng of astaxanthin per gram of muscle DW). These findings demonstrated that low‐inclusion microalgal additives in prawn diets over a short feeding period (4 weeks) can result in healthier adult prawn, and hence could contribute to the development of an optimized feeding strategy for prawn aquaculture close to harvesting.     

Effects of C:N ratio and substrate integration on periphyton biomass, microbial dynamics and growth of Penaeus monodon juveniles

An outdoor growth trial was conducted for 75 days to investigate the effect of C:N ratio and bamboo substrate (S) in brackish water shrimp culture. Penaeus monodon juveniles (0.35 ± 0.03 g) were stocked in experimental tanks with and without bamboo substrate. C:N ratio of 10 and 20 was manipulated with shrimp feed containing 32 % crude protein and rice flour as carbohydrate source. Addition of substrate and higher C:N ratio (CN20+S) significantly reduced (p < 0.01) inorganic nitrogen NH₃–N by 48.2 %, NO₃–N by 41.6 %, NO₂–N by 42.7 % compared with CN10. Substrate addition significantly (p < 0.05) reduced the turbidity by 22.4 % (CN10+S against CN10) and 20.7 % (CN20+S against CN20). Periphyton biomass and total heterotrophic bacterial load were significantly higher (p < 0.05) in CN20+S in comparison with CN10+S treatment. The autotrophic index increased from 123.4 ± 0.62 to 158.9 ± 3.8 for CN10+S and, 121.8 ± 1.6 to 228 ± 9.11 for CN20+S system. Provision of substrate and carbohydrate addition resulted in the highest body weight, 4.87 ± 0.12 g in CN20+S against 3.66 ± 0.07 g in CN20 and 2.90 ± 0.12 g in CN10 through provision of natural food in the form of periphytic algae and heterotrophic bacterial community. Treatment with substrates showed a higher survival rate by 7 % in comparison with without substrate treatments as it acted as shelter to shrimp during molting and there by reduced cannibalism. Thus, our results demonstrated that high C:N ratio and substrate addition improved growth, reduced FCR and better water quality conditions.     

Enhancement of growth performance, digestive enzyme activities and disease resistance in black tiger shrimp (Penaeus monodon) postlarvae by potential probiotics

Outbreaks of multidrug-resistant pathogens have been constraint on aquaculture production in Thailand, thereby controlling shrimp pathogens by preventive probiotics being importance to sustain the aquaculture system. In this study, the effect of potential probiotics, Bacillus subtilis and Enterococcus sp. was related to growth, digestive enzyme activities (trypsin and chymotrypsin) and pathogenic resistance by postlarval black tiger shrimp (Penaeus monodon). The experiment was divided into two treatments, in triplicate, with and without supplementation of probiotics as a food additive. Shrimp fed with probiotics for a culture period of 84 days showed a significant increase (P < 0.05) in weight (2.03 ± 0.29 g) and survival (71.91 ± 3.15 %) in comparison with non-treated shrimp (1.53 ± 0.28 g and 65.20 ± 5.68 %, respectively). Trypsin activity of treated shrimp was significantly higher (P < 0.05) than that of control shrimp whereas chymotrypsin activities of the two treatments were not significantly different (P > 0.05). After challenging P. monodon postlarvae with a shrimp pathogen, Vibrio harveyi for 10 days, percentage of mortality of P. monodon postlarvae fed with probiotics was 46.67 ± 1.44 %, significantly lower (P < 0.05) than that of non-treated shrimp (61.67 ± 6.29 %). This study showed that potential probiotics was appropriate for application in P. monodon postlarvae cultivation under laboratory condition due to improvement of shrimp weight and survival, enhancement of trypsin activity and reduced mortality causing by pathogenic V. harveyi. This is the first publication reported the effect of probiotics on trypsin and chymotrypsin activities of P. monodon postlarvae.     

Spermatophore cryopreservation and artificial insemination of black tiger shrimp, Penaeus monodon (Fabricius)

To develop an appropriate cryopreservation protocol for spermatophores of black tiger shrimp, Penaeus monodon, three cryoprotectants (dimethyl sulphoxide (DMSO), methanol (MeOH) and ethylene glycol (EG)) at two concentrations (5% and 10%) were examined. Artificial implantation of spermatophores was also carried out to assess the fertilizing ability of fresh and post‐thaw spermatophores. Spermatophores were collected during consecutive regenerations (15‐day intervals) and assessed for qualitative and quantitative changes and also for fertilizing ability by implantation. The mean fertilization rate for artificial insemination using post‐thaw spermatophore was 79.9±3.7%, lower than the fertilization rates observed for artificial implantation using fresh spermatophore and natural mating. Mean hatch rates for fresh spermatophore, frozen‐thawed spermatophore and natural mating were 88.8±0.6%, 87.8±0.4% and 88.3±0.5%, respectively; and there was no difference among the three groups. The mean fertilization rate of spermatophores collected during the first stripping was higher (90.6±0.6) than during the second stripping (85.7±2.6), but the mean hatch rate was not different between the two strippings. The highest mean sperm viability (79.7±0.4%) was obtained from DMSO (5%), with no survival observed in the 10% MeOH treatment. Spermatophore weight, total sperm count and percentage of abnormal sperm were not different between spermatophores collected at the first and second stripping. This is the first study to report high fertilization and hatch rates from cryopreserved spermatophore using artificial implantation of spermatophore before spawning.