Aspergillus niger fermented plant protein mix as a potential substitute for fishmeal in the diet of Penaeus vannamei (Boone, 1931)

A 45‐day feeding trial was conducted to evaluate the utilization of fermented plant protein mix (FPPM) as a fishmeal substitute in the diet of Penaeus vannamei. Six isonitrogenous and isolipidic diets were formulated by substituting fishmeal at 0% (control), 50%, 60%, 70%, 80% and 90% (FR‐0, FR‐50, FR‐60, FR‐70, FR‐80 and FR‐90, respectively) using FPPM. Shrimps fed with diets FR‐50, FR‐60 and FR‐70 had no significant difference in weight gain (173.22%–178.07%) with the control group (179.50%). However, the broken‐line regression indicated that 70.8% fishmeal could be substituted by FPPM. The trend of digestibility was reflected in growth performance. Survival had no significant difference among the treatments. Fishmeal substitution had significant (p < .05) variations in protease and amylase activity, but not in lipase. Except lipid, other body composition was not affected due to dietary change. A decrease (p < .05) in haemolymph total protein, cholesterol and triglycerides was observed to increase in fishmeal substitution. A higher fishmeal substitution (FR‐90) had significantly (p < .05) lower total haemocyte count and the reverse was true for phenoloxidase. Results conclude that a combination of fermented ingredients would be more effective rather using alone in shrimp feed, with a great potential in reducing the pressure on fishmeal.     

Effect of fungal fermentation on apparent digestibility coefficient for dry matter,crude protein and amino acids of various plant protein sources in Penaeus vannamei

Four commercial solvent‐extracted oilseed meals/cakes, viz. soybean meal (SBM), groundnut oil cake (GNC), rapeseed meal (RSM) and sunflower oil cake (SFC), were fermented with fungus Aspergillus niger, and its effect on apparent digestibility coefficient (ADC) was studied in Penaeus vannamei. Reference diet and eight experimental diets containing 700 g/kg reference diet and 300 g/kg test ingredient with 5 g/kg chromic oxide were formulated. Each diet was randomly allotted to three tanks containing ten shrimps. Shrimps were adapted to the experimental diets for a week, and faeces were collected using Falcon tube from second week onwards. The ADC of all the ingredients significantly (p < .05) increased with fermentation and the increase being higher in SBM (78.46%–91.71%) for dry matter and in SFC (71.51%–87.02%) for protein. Analysis of variance showed that the ADC of both dry matter and protein significantly (p < .05) differed in treatments (p = <.001) and ingredients (p = <.001). The average ADC of ingredients was ranked as SBM > GNC > RSM > SFC. The most digestible essential amino acid (EAA) in fermented ingredients was methionine in SBM, arginine in GNC, valine in RSM and histidine in SFC. A better improvement in amino acid digestibility was observed in fermented SFC. Results indicated that P. vannamei efficiently digests fermented ingredients compared to unfermented ones.     

Apparent digestibility coefficients of fungal fermented plant proteins in two different penaeid shrimps—A comparative study

Using the 70:30 replacement method and chromium as an inert marker, the digestibility of four fermented oilseed meals/cakes (soybean meal (FSBM), groundnut oil cake (FGNC), rapeseed meal (FRSM), and sunflower oil cake (FSFC)) were determined in Penaeus monodon and Penaeus indicus. Apparent dry matter digestibility (ADMD) of fermented ingredients was ranked as FSBM > FGNC > FRSM > FSFC. The critical variations in apparent protein digestibility (APD) in P. monodon (0.816%) and P. indicus (0.608%) were lower than the ADMD. Appaprent amino acid digestibility (AAD) was >90% in FSBM for both the species and was lower for other ingredients. Protein had a higher digestibility than total amino acids and was in the range of 0.69%–2.71% in P. monodon and 0.32%–2.75% in P. indicus. A correlation between the ADC of total amino acids and protein was found to be r = 0.8229 in P. indicus and r = 0.7447 in P. monodon. Data were further subjected to two way analysis of variance for assessing the digestibility variations between the species. It was observed that P. indicus had higher values of ADMD than P. monodon in FSBM (2.97%) and FRSM (1.22%) and the reverse was true in FGNC and FSFC. The APD was high in P. indicus for FSBM, FGNC and FRSM but not for FSFC. However, significant variations could be noticed in AAD between the species.     

Optimization of fixation methods for image analysis of the hepatopancreas in whiteleg shrimp,Penaeus vannamei (Boone)

Pathology in penaeid shrimps relies on histology, which is subjective, time‐consuming and difficult to grade in a reproducible manner. Automated image analysis is faster, objective and suitable for routine screening; however, it requires standardized protocols. The first critical step is proper fixation of the target tissue. Bell & Lightner's (A Handbook of Normal Penaeid Shrimp Histology, 1988, The World Aquaculture Society, Baton Rouge) fixation protocol, widely used for routine histology of paraffin sections, is not optimized for image analysis, and no protocol for frozen sections is described in the available literature. Therefore, the aim of this study was to optimize fixation of the hepatopancreas (HP) from whiteleg shrimp (Penaeus vannamei) for both paraffin and frozen sections using a semiquantitative scoring system. For paraffin sections, four injection volumes and three injection methods were compared, for frozen sections, four freezing methods and four fixation methods. For paraffin sections, optimal fixation was achieved by increasing threefold the fixative volume recommended by Bell and Lightner, from 10% to 30% of the shrimp body weight, combined with single injection into the HP. Optimal fixation for frozen sections was achieved by freezing the cephalothorax with liquid nitrogen, followed by fixation of the section with 60% isopropanol. These optimized methods enable the future use of image analysis and improve classical histology.     

Partial replacement of fishmeal with biofloc meal in the diet of postlarvae of the Pacific white shrimp Litopenaeus vannamei

This study evaluated the potential for the replacement of fishmeal (FM) by biofloc meal (BM) in the diet of Litopenaeus vannamei postlarvae. Four isonitrogenous (437.42 g kg^?1) and isocaloric (19.84 kJ g^?1) diets were formulated, in which FM was replaced with BM at 0% (T0), 7.5% (T7.5), 15% (T15) and 30% (T30). A commercial diet (CD) was used as reference. The study was conducted over 42 days in 50 L tanks connected to a water recirculation system. All tanks were stocked with three postlarvae (initial weight 0.0023 g) per litre. Shrimp survival, weight gain (WG), final weight (FW), specific growth rate (SGR) and protein efficiency ratio (PER) were measured. Survival rates were high (>91.1%) in all treatments. As the T30 treatment returned a significantly (<0.05) better performance in WG, FW, SGR and PER than all other treatments, partial replacement of FM with BM does not appear to affect productivity. In fact, a regression analysis indicated that a FM to BM replacement level of over 20% may actually improve shrimp growth. In addition to providing a cost‐effective alternative ingredient for L. vannamei postlarvae diets, the production of meal also represents a more sustainable way of disposing of the excess of biofloc produced by BFT systems.     

Effects of a partitioned aquaculture system on water quality and growth of Penaeus vannamei

The effects of system partition on the water quality were studied using three shrimp ponds (Ts), and each one of the shrimp pond was connected individually with a fish pond by piping and pump. Three other isolated shrimp ponds were designated as the Cs treatment. The results showed that the implementation of the system can improve the water quality in the shrimp ponds. Particulate organic matter (POM), chemical oxygen demand (COD), nitrite nitrogen (NO₂–N) and nitrate nitrogen (NO₃–N) in the Ts were lower than those in the Cs during the experiment, and in some cases, they were significantly lower (p < 0.05). Throughout most of the experimental period, the soluble reactive phosphorus in the Ts treatment were greater than that in the Cs treatment (p < 0.05), and N/P ratio of the water column was lower in the Ts treatment than in the Cs treatment throughout most of the experimental period (p < 0.05). These results indicate that the partitioned system contributed to the stability of water quality and N/P ratio in the water. Vibrio parahaemolyticus was dominant in the third to seventh weeks in Cs, and the dominant condition was found only in seventh week in Ts. The result indicated that system may contribute to the slowing down of the breakout of V. parahaemolyticus in the shrimp pond. According to the redundancy analysis, COD, chlorophyll a and POM were mainly positively related to the dominance index of V. parahaemolyticus, and maybe POM can serve as an indicator to reflect the trend of V. parahaemolyticus.     

Effect of starvation and refeeding on the hepatopancreas of whiteleg shrimp Penaeus vannamei (Boone) using computer‐assisted image analysis

Under normal farming conditions, shrimp can experience starvation periods attributable to disease outbreaks or adverse environmental conditions. Starvation leads to significant morphological changes in the hepatopancreas (HP), being the main organ for absorption and storage of nutrients. In the literature, limited research has described the effect on the HP of periods of starvation followed by refeeding and none in whiteleg shrimp (Penaeus vannamei) using computer‐assisted image analysis (CAIA). This study describes the effect of starvation and starvation followed by refeeding on the HP of whiteleg shrimp using CAIA. Visiopharm^® software was used to quantify the following morphological parameters, measured as ratio to the total tissue area (TLA): total lumen area (TLA:TTA), haemocytic infiltration area in the intertubular spaces (HIA:TTA), B‐cell vacuole area (VBA:TTA), lipid droplet area within R cells (LDA:TTA) and F‐cell area (FCA:TTA). Significant changes were measured for HIA:TTA and LDA:TTA during starvation (increase in HIA:TTA associated with decrease in LDA:TTA) and starvation followed by refeeding (decrease in HIA:TTA associated with increase in LDA:TTA). In the future, HIA:TTA and LDA:TTA have the potential to be used in a pre‐emptive manner to monitor the health of the HP, facilitate early diagnosis of diseases and study the pathophysiology of the organ.     

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.     

Thermal challenge of the Indian shrimp Penaeus indicus

This study investigated short‐term effects of increasing water temperature from 27 to 41°C on survival and feed consumption of Penaeus indicus at three different ages: PL₂₅ (postlarvae 25 days old), PL₅₀ and PL₉₀. For each age group, water temperature was maintained at 27°C in the control, but increased to 32, 35, 38 and 41°C at a rate of 1°C every eight hours. The temperature was then kept stable until the end of the 7‐day experiment. Results showed that increasing water temperature affected both survival and feed consumption of the experimental shrimps (p < .01). Survival was highest at 32 and 35°C ranging from 93.8% to 100%, but significantly reduced to 40.0%–81.6% at 38°C. No shrimp survived the 41°C treatment. PL₂₅ were more tolerant to 38–41°C than PL₅₀ and PL₉₀ in terms of survival. Increasing water temperature had no effects on feed consumption of PL₂₅ (p > .05). For PL₅₀ and PL_90, feed consumption significantly increased at 38 and 41°C (p < .01) and was similar within the range of 27–35°C. This study suggests that P. indicus in tropical areas can tolerate water temperatures of at least 35°C and should be considered for farming during the summer time.     

Effects of dietary administration of Macsumsuk® on growth and stress to low salinity and low dissolved oxygen in whiteleg shrimp,Litopenaeus vannamei (Boone, 1931) juveniles

An 8‐week study was conducted to explore the results of Macsumsuk^® as a feed additive on the stress tolerance and growth of Litopenaeus vannamei in 15 culture tanks of 36 L each. Three hundred shrimp averaging 0.1 ± 0.01 g were fed with five isonitrogenous (48.38 ± 0.38% CP) diets (in triplicate groups) containing kaolinite (Macsumsuk^®) at 0%, 0.3%, 0.6%, 1.2% and 2.4%, namely Mk₀, Mk_0.3, Mk_0.6, Mk_1.2 and Mk_2.4. Specific growth rate (SGR) and weight gain (WG) of shrimp fed diets Mk_1.2 and Mk_2.4 were significantly better than those of shrimp fed diet Mk₀ (p < .05). However, SGR and WG of shrimp fed diets Mk_0.6, Mk_1.2 and Mk_2.4 were not significantly different. Protein efficiency ratio (PER) and feed efficiency (FE) of shrimp fed diets Mk_1.2 and Mk_2.4 were significantly better than those of shrimp fed diets Mk₀, Mk₀.₃ and Mk_0.6. Furthermore, the survival of shrimp fed diet Mk_2.4 was significantly lower than that of shrimp fed diet Mk_0.6 (p < .05). Cumulative mortality of shrimp fed diet Mk_1.2 was significantly lower than that of shrimp fed diet Mk₀ at 1–1.5 hr post‐stress to low dissolved oxygen (from 6.1 mg/L to 2.9 mg/L) and 4–5 hr post‐stress to low salinity (from 32‰ to 1‰) (p < .05). The optimum dietary Macsumsuk^® level for juvenile L. vannamei was determined as 1.97% by the polynomial regression analysis of weight gain.     

Partial replacement of fishmeal by fermented soybean meal in diets for juvenile white shrimp (Litopenaeus vannamei)

As an important feed ingredient, fermented soybean meal (FSM) has been widely used in aquatic animals due to its stable sources and reasonable price. Here, we evaluated the potential for replacement of fishmeal (FM) with FSM in diet of Litopenaeus vannamei. Five isonitrogenous (410 g/kg) and isolipidic (80 g/kg) diets were formulated: a control diet containing 320 g/kg FM and four experimental diets in which FM in control diet was replaced by FSM at 10 (FSM10%), 20 (FSM20%), 30 (FSM30%) and 40% (FSM40%). An 8‐week feeding trial was conducted in fifteen fibreglass tanks with 50 shrimps per tank. After 8 weeks trial, FSM20% had significantly enhanced growth performance (p < 0.05). No significant difference was found in body composition and digestive enzyme activities of all groups (p > 0.05). Through real‐time quantitative PCR analysis, tor and s6k expression levels of FSM20% were significantly up‐regulated (p < 0.05). Results of western blot showed that the phosphorylation of S6K was not significantly affected by different dietary treatments (p > 0.05), which suggested mTOR signalling pathway was not affected by FSM diets. Based on the above data, 20% replacement of FM with FSM was reasonable and advantageous for L. vannamei diet.     

Nutritional evaluation of fermented Jatropha protein concentrate in Labeo rohita fingerlings

Jatroha protein concentrate (JPC) was prepared by iso‐electric precipitation method. A 45‐day feeding trial was conducted for the nutritional evaluation of JPC and fermented Jatropha protein concentrate (FJPC) in Labeo rohita fingerlings. Seven iso‐nitrogenous (340 g kg^?1 CP) and iso‐energetic (17 MJ kg^?1) diets such as C (soy protein concentrate⁾, T₁ (50 g kg^?1 JPC), T₂ (100 g kg^?1 JPC), T₃ (200 g kg^?1 JPC), T₄ (50 g kg^?1 FJPC), T₅ (100 g kg^?1 FJPC) and T₆ (200 g kg^?1 FJPC) were fed to respective groups. The weight gain % and protein effciency ratio showed a decreasing trend with higher inclusion of JPC whereas these parameters increased gradually on higher inclusion of FJPC. FCR showed the reverse trend. The protease, GOTand GPT activities of FJPC fed groups were significantly higher than their non‐fermented counterparts whereas a reverse trend was observed for LDH and MDH enzyme activities in muscle and liver. The survival rate was 100% in all the treatment groups except for T₂ and T₃. Phorbol esters (PEs) in FJPC was non‐detectable. Results suggested that solid state fermentation with Aspergillus niger is an effective method for removal of PEs and FJPC is a promising protein source in aquafeed.     

Evaluation of ultra‐low gossypol cottonseed and regular glandless cottonseed meals as dietary protein and lipid sources for Litopenaeus vannamei reared under zero‐exchange conditions

The current study evaluated transgenic cotton lines with normal levels of gossypol/terpenoids in the vegetative and floral tissues, but with ultra‐low gossypol in the seeds as a replacement for glandless cottonseed meal (GCSM) and fishmeal. A 64‐day growth trial evaluated the ability of cottonseed meals from a natural glandless cotton variety/mutant, two transgenic Ultra‐low Gossypol Cottonseed (ULGCS) lines, a non‐transgenic parental control and a commercial variety, to replace 355 g kg^?1 fishmeal in a diet containing 350 g kg^?1 crude protein. Juvenile Litopenaeus vannamei (1.48 ± 0.29 g) were stocked (40 shrimp m^?3) with six replicates. No significant differences were found between all formulated diets in terms of final weight, survival and feed conversion ratio. The commercial cottonseed variety displayed a significantly lower feed efficiency ratio and protein efficiency ratio than one of the ULGCS diets. These results suggest that GCSM and/or transgenic ULGCS meals can be used to replace fishmeal in commercial shrimp diets.     

Nutritional evaluation of an improved soybean meal as a fishmeal replacer in the diet of Pacific white shrimp,Litopenaeus vannamei

A 12‐week growth trial was conducted to assess an improved soybean meal (ISBM) in the diet of Pacific white shrimp (PWS). Four isonitrogenous and isolipidic experimental diets were formulated: the control diet (ISBM‐0) contained 30% fishmeal (FM) and three test diets (ISBM‐33, ISBM‐66 and ISBM‐100) were formulated by gradually replacing FM on an isonitrogenous basis. Groups of PWS (3.2 g mean initial weight) were fed their respective experimental diets three times per day. Production performance of PWS was unaffected by diet, except the final weight of shrimp‐fed ISBM‐66 diet was significantly lower than those fed ISBM‐0. Whole‐body moisture was significantly higher and protein was significantly lower in PWS‐fed ISBM‐100 versus shrimp‐fed ISBM‐0. PWS‐fed ISBM‐66 had significantly higher oleic acid than shrimp‐fed ISBM‐0 and linolenic acid versus shrimp‐fed ISBM‐0 and ISBM‐100. The lowest linoleic acid and n‐6 polyunsaturated fatty acids levels were found in shrimp‐fed ISBM‐100. Based on second‐degree polynomial regression analysis of final weight and weight gain, the estimated optimum levels of ISBM to replace FM in PWS diets are 89.13% and 95.56% respectively. Overall, results indicate alternative, sustainable, plant‐based protein sources such as ISBM may potentially replace FM in PWS diets.     

Partial replacement of fish meal with peanut meal in practical diets for the Pacific white shrimp,Litopenaeus vannamei

In this study, we replaced fish meal with peanut meal (PM) in isonitrogenous and isolipidic diets for Pacific white shrimp at inclusion levels of 0, 70, 140, 210, 280 and 350 g kg^?1. The diets were hand‐fed to three independent groups of shrimp three times a day over a 6‐week period. Shrimp fed PM diets at a level of 280 g kg^?1 or higher had lower per cent weight gain compared with those fed the basal diet, whereas shrimp fed PM diets at 140 g kg^?1 or higher had a lower feed utilization and protein efficiency ratio compared with shrimp fed the basal diet. The feeding rate in shrimp fed PM diets at 350 g kg^?1 and the survival and protease activity in shrimp fed PM diets at 210 g kg^?1 or higher were lower than that in shrimp fed the basal diet. Diets containing 280 g kg^?1 or higher of PM caused an increase in the whole‐body moisture content of the shrimp, but decreased whole‐body protein and ash contents compared with the basal diet. Nutrient digestibility was lower or tended to be lower in shrimp fed a PM diet compared with those fed the basal diet. The activities of peroxidase and acid and alkaline phosphatases in plasma decreased with increasing levels of PM inclusion up to 210 g kg^?1. Superoxide dismutase activity decreased at dietary PM levels of 280 g kg^?1 or higher. Aflatoxin B₁ residue in the muscle was not affected by any of the treatments and remained low. The data suggest that up to 140 g kg^?1 of PM could be included in practical diets for Pacific white shrimp.     

Effects of replacing fishmeal with fermented and non‐fermented rapeseed meal on the growth,immune and antioxidant responses of red sea bream (Pagrus major)

The effect of rapeseed meal (RM) and Aspergillus oryzae fermented rapeseed meal (RM‐Koji) on red sea bream (Pagrus major) was examined. Three groups of fish (initial weight, 4.5 ± 0.02 g) were fed a basal diet (RM0) and two test diets where half of fishmeal was replaced by RM (RM50) and RM‐Koji (FRM50) for 56 days. The obtained results showed that fish fed RM0 and FRM50 exerted significantly higher growth performance, feed utilization and haemoglobin level but lower triglyceride and cholesterol than RM50 group (p < 0.05). Interestingly, except of antiprotease activity, all the immune parameters including lysozyme, respiratory burst (NBT) and bactericidal activities were significantly increased in fish fed RM0 and FRM50 diets compared to RM50 diet (p < 0.05). In addition, malondialdehyde and reactive oxygen metabolites were significantly reduced in RM0 and FRM50 groups over RM50 group (p < 0.05). The present results suggest that fermented RM induced better growth performance and immune responses than feeding red sea bream with non‐fermented RM and both RM and RM‐Koji improved the antioxidative status of fish, making RM‐Koji an interesting candidate as a functional feed for aquatic animals.     

Replacement of fish oil with a DHA‐rich Schizochytrium meal on growth performance,activities of digestive enzyme and fatty acid profile of Pacific white shrimp (Litopenaeus vannamei) larvae

This trial was conducted to evaluate the effects of replacing dietary fish oil with Schizochytrium meal for Pacific white shrimp (Litopenaeus vannamei) larvae (initial body weight 4.21 ± 0.10 mg). Six test microdiets were formulated using Schizochytrium meal to replace 0 g/kg, 250 g/kg, 500 g/kg, 750 g/kg, 1000 g/kg or 1500 g/kg fish oil DHA. No significant differences were observed in survival, growth, final body length and activities of digestive enzyme among shrimp fed different diets (p > .05). No significant differences were observed in C20:5n‐3 (EPA) in muscle samples (p > .05). C18:3n‐3 and C20:4n‐6 in muscle increased as Schizochytrium meal replacement level increased (p < .05). No significant differences were observed in C22:6n‐3 (DHA) and n‐3 fatty acids among shrimp fed diets that algae meal replaced 0 g/kg ‐ 1000 g/kg of fish oil. Shrimp fed diet R150 had higher DHA content than other groups and had higher n‐3 fatty acids than that of shrimp fed diets R50, R75 and R100 (p < .05). C18:2n‐6, PUFA and n‐6 fatty acids in muscle increased, while n‐3/n‐6 ratio decreased with increasing algae meal replacement level from 0 g/kg to 1000 g/kg (p < .05). In conclusion, Schizochytrium meal could replace 1500 g/kg fish oil DHA in the microdiets without negatively affecting shrimp larvae survival, growth and activities of digestive enzyme.