Evaluation of Agave lechuguilla by‐product crude extract as a feed additive for juvenile shrimp Litopenaeus vannamei

Agave lechuguilla is a succulent plant species, mainly distributed in the northeast of Mexico and south of the United States of America. The main use for this plant is the fibre´s extraction (known as Tampico fibres), resulting in 15% of fibres and 85% of a by‐product waste named guishe. The lechuguilla collectors, normally incinerate the guishe, thus causing environmental contamination. Interestingly, recent studies showed that guishe contains molecules with nutritional properties, such as saponins, flavonoids and sugars. Therefore, in this work, we evaluated the effect of the crude extract of guishe as a feed additive in whiteleg shrimp diets. According to that, MS‐HPLC analysis of the extract showed the presence of saponins such as diosgenin, smilagenin, hecogenin, manogenin, tigogenin hexose, yucagenin, chlorogenin, diosgenin diglucoside and the flavonol, quercetin. After chemical analysis, the crude extract was included into an experimental diet in four levels; 0% (L0%), 0.1% (L0.1%), 0.3% (L0.3%) and 0.6% (L0.6%). Dietary incorporation of the extract was evaluated by zootechnical performance, haemolymph biochemistry, histomorphology and digestive enzyme activity of shrimps. After 5‐week feeding, the L0.3% diet showed significantly higher growth and better feed utilization among treatments. A significant increase in tubule epithelium height and tubule coverage area from hepatopancreas in shrimp under L0.3% diet compared with the control diet suggest an improvement of the health and nutritional status of the shrimp. Inclusion of L0.3% and L0.6% of the crude extract resulted in a reduction in amylase activity, without effect in glucose levels in the haemolymph. Thus, we suggest that lechuguilla guishe crude extract contains nutritional molecules that may be used as a feed additive to promote shrimp productivity.     

Evaluation of dietary chitosan effects on growth performance,immunity, body composition and histopathology of Nile tilapia (Oreochromis niloticus) as well as the resistance to Streptococcus agalactiae infection

The present investigation aimed to evaluate the effect of dietary chitosan supplementation on growth performance, body composition, immune response and histopathology of Nile tilapia, and also the in vitro antibacterial activity of chitosan against Streptococcus agalactiae (S. agalactiae). About 180 fish (average body weight 39.3 ± 0.3 g) were randomly divided into three groups according to chitosan supplementation: control group (basal diet without chitosan), Ch3 group (3 g chitosan/kg diet) and Ch5 group (5 g chitosan/kg diet). Growth performance parameters and body proximate composition were measured before infection but biochemical parameters and lysozyme and antibacterial activities before and after experimental infection. Results of the present investigation showed dietary chitosan (5 g chitosan/kg diet) significantly (p < .05) improved growth performance parameters, body composition (dry matter, crude protein, ether extract, ash, and carbohydrate) and serum biochemistry (total protein, albumin, globulin, with no effect on AST, ALT, urea and creatinine) before infection in Ch5 group than the control. After infection, liver enzymes (serum AST and ALT) were maintained lower in fish fed Ch3 or Ch5 than the control. Serum lysozyme and bactericidal activities significantly increased (p < .05) in chitosan groups before and after the challenge. The mortality rate was markedly reduced in the Ch3 group and prohibited in the Ch5 group after the experimental infection. In conclusion, feeding 3 or 5 g chitosan/kg diet increased the growth rate and improved FCR of Nile tilapia. In addition, it reduced mortality by its antibacterial and immunostimulant effects.     

Evaluation of limb autotomy as a promising strategy to improve production performances of mud crab (Scylla olivacea) in the soft‐shell farming system

Soft‐shell crab, a newly moulted crab having non‐calcified and hydrated soft exoskeleton, farming has been increasing tremendously in recent years throughout the world due to its high market price and consumer preferences. However, high aggressiveness and cannibalistic behaviour of mud crab limit its culture to an individual chamber system which ultimately makes the soft‐shell mud crab farming space and labour‐intensive. Therefore, the aim of this study was to improve the efficiency of soft‐shell crab production by autotomy of the claw and/or walking legs to reduce the moulting duration and intensify the culture system through double stocking in the existing individual chamber culture system. To achieve our objectives, a series of experiment were conducted at both indoor and on‐farm levels by removing the claw and/or walking legs of Scylla olivacea for two consecutive moulting cycles. Results showed that both claw ablation significantly (p < .01) shortened the moulting duration and increased the carapace width, weight gain, specific growth rate, feeding efficiency and yield parameter compared to the non‐ablated crabs more prominently at 1st moulting than the 2nd moulting in both indoor and on‐farm trials. Ablation of all walking legs while keeping the claw intact did not significantly influence (p > .05) the moulting duration and carapace width, but significantly (p < .01) improved the specific growth rate, weight gain, feeding efficiency and yield parameters at 1st moulting compared to the non‐ablation and unilateral three walking leg ablation groups. However, ablation of all appendages (both claw and walking legs) showed significant (p < .01) improvements of carapace width, weight gain, specific growth rate, feeding efficiency and yield parameters, and shortened the moulting duration up to 23 days from 34 days during 1st moulting and up to 27 days from 40 days at 2nd moulting. The survivals of different autotomized mud crabs were statistically non‐significant (p > .05) to the non‐autotomized crabs in various indoor and on‐farm trials. Besides improving the production performances, ablation of bilateral claw and all appendages allowed to double the stocking density in the existing individual chamber‐based soft‐shell crab farming that enhanced the production about threefold higher than the existing individual chamber‐based culture system. Finally, our results suggest that autotomy could be a promising technology for the commercial farming of soft‐shell mud crab in near future.     

Potential bioavailability of protein and lipids in feed ingredients for the three‐spot cichlid Amphilophus trimaculatus: An in vitro assessment

For the commercial culture of the three‐spot cichlid (Amphilophus trimaculatus), it is important to have available a set of protein and lipid ingredients suitable for feed manufacturing, which allow to optimize the culture at affordable costs. The in vitro digestibility was evaluated with pH‐stat technique, using stomach and intestine multi‐enzymatic extracts from A. trimaculatus juveniles. The digestion of protein ingredients was taken place both in acid and in alkaline conditions, quantifying the total of free amino acids at the end of the hydrolysis, whereas the digestion of lipid ingredients was taken place only in alkaline conditions. The digestive enzymes of this species showed a better affinity to raw materials of animal origin such as beef meal, fish meal and premium chicken meal, which presented better digestibility. The best lipid ingredient that could be used is the fish oil. However, it is possible to use ingredients from the plant sources, such as soy or cereals, and vegetable oils such as olive, corn or soy oil. It is necessary to complement this study in vitro with the digestibility analysis in vivo, in order to determine the inclusion percentages in feeds and costs and therefore to develop an efficient feed formulation for the aquaculture of A. trimaculatus.     

Minimally invasive evaluation of the anaesthetic efficacy of MS‐222 for ornamental discus fish using skin mucus biomarkers

Skin mucus has been demonstrated to provide stress biomarkers for evaluating the physiological status, providing new convenient and non‐invasive methods to detect stress response in fish. Here, we investigated the anaesthetic efficacy of tricaine methanesulphonate (MS‐222; 75–115 mg/L) for discus Symphysodon aequifasciata (34.27 ± 4.46 g; 8.10 ± 0.59 cm) using skin mucus stress biomarkers. The induction time, recovery time and respiratory frequency were also determined. According to the criteria for anaesthesia and recovery, discus fish to reach stage A3 (deep anaesthesia) within 3 min and to reach stage R4 (full recovery of normal behaviour) within 5 min were observed at 95–105 mg/L MS‐222. Respiratory frequency increased first and then decreased during MS‐222 exposure and increased after recovery. At 10 min after deep anaesthesia, a lower mucus glucose was only observed at 115 mg/L MS‐222. No change in mucus cortisol and increased lactate were observed in all treatments. Increased mucus protein was observed at 75, 85 and 95 mg/L MS‐222. At 10 min after recovery, increased mucus glucose and decreased mucus protein were observed at 85, 95 and 115 mg/L MS‐222, but increased mucus cortisol only at 115 mg/L and lactate only at 75 and 105 mg/L MS‐222. At 24 hr after recovery, mucus glucose returned to the initial level only at 75, 95 and 105 mg/L MS‐222, while cortisol at 75 and 85 mg/L and protein and lactate at 75 mg/L respectively. Overall, the effective dose of MS‐222 for discus fish has been suggested to be 95–105 mg/L.     

Enhancing rotifer Brachionus calyciflorus population growth with commercial probiotics

The effect of a commercial probiotic (NanoCrusta, Altacrusta, Mexico City, Mexico) on the growth performance of Brachionus calyciflorus Pallas, 1766, was evaluated. In a first approach, probiotics were supplied in four densities (2.0 × 10³, 1.1 × 10⁵, 2.1 × 10⁵ and 2.1 × 10⁶ cells/ml), alone and in combination with Chlorella vulgaris (1 × 10⁶ cells/ml). The test rotifer did not grow on the probiotic alone. However, when probiotics + C. vulgaris were added, the maximum densities (D_max; ind/ml) and population growth rates (r) observed were higher. In the second experiment, probiotics were supplied at five higher densities (2.1 × 10⁶, 4.2 × 10⁶, 8.5 × 10⁶, 1.7 × 10⁷ and 3.4 × 10⁷ cells/ml) with C. vulgaris and a control treatment with only C. vulgaris (probiotic‐free). Treatments supplied with probiotics between 2.1 × 10⁶ and 1.7 × 10⁷ cells/ml showed significantly higher D_max and r than the control treatment. The results showed a positive effect of probiotic bacteria when supplied with C. vulgaris. The best outcome showed a D_max 2.16 times and an r 1.63 times higher than the density of the control treatment. Growth rates were higher in the treatments with probiotics compared to the control. We conclude that application of NanoCrusta is feasible to improve B. calyciflorus production, but the effects need to be tested in larger scales.     

Assessment of dietary lipid sources in tropical gar,Atractosteus tropicus larvae: Growth parameters and intermediary lipogenic gene expression

Five experimental diets containing different lipid sources, fish oil (D1), soybean lecithin (D2), corn oil (D3), canola oil (D4) and olive oil (D5), were evaluated in Atractosteus tropicus larvae for the relative gene expression of the enzymes fatty acid synthase (fas), acetyl‐CoA carboxylase 1 (acc1) and carnitine palmitoyltransferase 1C (cpt1c), in addition to their effects on larval growth, survival and cannibalism during a 30‐day feeding trial. Higher growth and survival were obtained in treatments D1 and D2, and lower performance in diets D3, D4 and D5. The highest levels of expression of fas and acc1 occurred in larvae fed with D1, which contained high amounts of n‐3 long‐chain polyunsaturated fatty acids (LC‐PUFA), mainly DHA and EPA FA are regulators of lipogenesis. The higher cpt1c expression in plant‐based diets is attributed to the fact that these diets are rich in α‐linolenic acid (ALA) and low DHA, EPA and ARA levels that favour ß‐oxidation. In conclusion, the diets with fish oil (D1) and soybean lecithin (D2) were the best treatments for larval growth, survival and cannibalism and thus appear to meet both lipid and energy requirements of A. tropicus larvae, meanwhile the use of vegetable oils influences the expression of intermediary lipogenic genes.     

Laboratory and on‐farm evaluation of low‐cost salt mixtures for use during salinity acclimation and the nursery phase of Pacific white shrimp (Litopenaeus vannamei)

The current study evaluates the efficacy of a low‐cost salt mixture (LCSM) to replace expensive reconstituted sea salt (RSS) in the salinity acclimation and nursery phase of Pacific white shrimp under laboratory and farm conditions. LCSM was formulated to yield sodium, potassium, calcium and magnesium concentrations closely comparable to that of diluted seawater. Laboratory‐based nursery trials were conducted at 2, 6 and 15 g/L salinities, incrementally replacing RSS with LCSM (25%, 50%, 75% and 100%) at four replicates per treatment. Thirty postlarvae were reared for 7 days in 24‐L aquaria during the 2 and 6 g/L trials, while the nursery trial for 15 g/L salinity was conducted for 21 days with 400 postlarvae stocked in 150‐L tanks. On‐farm evaluation of LCSM was carried out in two tank‐based systems installed on levees adjacent to shrimp production ponds. RSS was incrementally replaced with LCSM (0%, 50%, 75% and 100%) and 100 postlarvae stocked into each 800‐L tank. Salinity acclimation was done from 30 g/L to 6 or 1.5 g/L within 2–3 days by pumping water from adjacent shrimp production ponds. Following salinity acclimation, the S4 system maintained flow‐through at 1.5 g/L, while N10 system was maintained static at 6 g/L salinity. At the conclusion, no significant differences were observed for either survival or growth of shrimp postlarvae between RSS and LCSM treatments at all salinities examined. Results reflect the potential use of LCSM to replace RSS, which could be an excellent solution to bring down the cost of production in inland shrimp aquaculture.     

Development of a method for live octopus (Octopus vulgaris) transportation for long distance at high densities

The development of new octopus‐based products with a growing economic value ensures the commercial interest of this species, making live octopus export an activity of great interest. The aim of this study was to develop a method for long‐distance transportation of live octopus at high densities. The system was composed by 220‐L tanks with cooling and aeration, where the animals were kept separated from each others. The water temperature was maintained at 10°C, after a decreasing of 1°C/hr. Live octopus transportation was tested for 48 hr at two densities: 50 kg/m³ and 100 kg/m³. During this period, water parameters were monitored. Stress response was evaluated through the analysis of haemolymph, muscle and brain tissues. No mortality was registered after 48 hr for both treatments. In all trials, water quality remained within the normal limits in both densities; there was, however, a significative increase of ammonia levels in the water. Ammonia, dopamine and Hsp70 levels were analysed in the beginning and at the end of the experiment for both densities; however, no significant differences were found among them. In general, this system seems to be a viable solution for live octopus 48‐hr transportation at a density of 100 kg/m³.     

Development and testing of a Bayesian population model for the bigeye thresher shark,Alopias superciliosus,in an area subset of the western North Pacific

A Bayesian population modelling tool integrating separable virtual population analysis, per‐recruit models and age‐structured demographic analysis was developed for the bigeye thresher Alopias superciliosus (Lowe) population in an area subset of the western North Pacific. The mortality rates for years 1989–2016 were estimated, various biological reference points and associated risks of decline were also estimated, and alternative harvest strategies for the stock were evaluated. Estimates of the posterior mean of fishing mortality for bigeye thresher shark suggest fishing pressure has been high in recent years (2011–2016). The estimated population growth rate (λ) (without fishing) obtained from age‐structured demographic model was relatively low (λ = 1.01 per year; 95% confidence intervals of 1.00 and 1.03 per year). Risk analyses revealed that only low levels of fishing pressure (10% of the current fishing pressure) over a wide range of ages could maintain a relatively low risk of population decline for bigeye threshers. Sensitivity testing indicated that the model is robust to prior specification. The developed framework could be used as an assessment tool to evaluate the risk of decline for other widely distributed pelagic shark species where insufficient catch and effort data are available.