International study on Artemia. XVIII. The hatching rate of Artemia cysts — A comparative study

A comparative study has been carried out on the hatching rate of Artemia cysts of different geographical origin and on different batches of the same source. Substantial variation, among as well as within geographical strains, indicates that the hatching-rate criterion is not strain specific.

In an attempt to identify the parameters which influence the hatching rate, it was found that the cyst-drying technique as well as the storage conditions may significantly affect this criterion. It has also been demonstrated that the cysts' hatchability is influenced by their previous drying and storing conditions. Optimal hatching results are guaranteed when a fluidized bed dryer is used and the cysts are stored under vacuum or nitrogen.     

International study on Artemia: XIX. Hatching data for ten commercial sources of brine shrimp cysts and re-evaluation of the “hatching efficiency” concept

A comparative hatching study has been carried out with Reference Artemia Cysts as well as cysts from Argentina, Australia, Brazil, Canada, France, Italy, the People's Republic of China, the Philippines and the U.S.A. Hatching rate, percentage and efficiency vary considerably from one cyst source to another. These hatching criteria are, however, not strainspecific since significant variation is found among cyst batches from the same geographical origin.

The limitations of the “hatching efficiency” concept are discussed and a new criterion “hatching output”, i.e. the biomass of nauplii expressed in mg dry weight produced per gram cyst product, is proposed for evaluation of the hatching quality of Artemia cyst brands.

The hatching quality of Reference Cysts and cysts from Canada and Argentina can be significantly improved by incubation of the cysts at low salinity.

For almost all commercial sources the use of decapsulated cysts results in a significant increase of the hatching output.     

Enrichment of Artemia with free methionine

Different enrichment procedures of the free amino acid (FAA) methionine were tested for Artemia nauplii. A direct enrichment protocol (methionine dissolved in the culture water) was compared with liposome enrichment protocols that varied in their membrane composition. During 16 h of direct enrichment in 5.3 mM methionine, the nauplii increased their content of free methionine between 20 and 30 times compared to the unenriched control (43.1±1.2 and 68.4±3.8 pmol·nauplius⁻¹ in two separate experiments vs. 2.4±1.0 pmol·nauplius⁻¹ in control). However, by encapsulating the identical amount of methionine into liposomes made from pure egg yolk phosphatidylcholine (PC) (>99% PC) and cholesterol, the nauplii content of free methionine reached 148.8±27.6 pmol·nauplius⁻¹, which is approximately 60 times more than in the unenriched control. Another liposome composition tested, made from crude egg yolk PC (>60% PC) and cholesterol, resulted in 90.5±4.1 pmol·nauplius⁻¹. The enriched nauplii still retained 80% of the free methionine after 8 h of incubation at conditions simulating feeding for Atlantic halibut larvae (13°C, 33.5 g·l⁻¹).

In conclusion: (1) Artemia nauplii can successfully be enriched with free methionine, (2) the high retention of free methionine in the Artemia nauplii following transfer to fish tanks shows that it is possible to offer fish larvae a feed with a high level of FAA, based on enrichment of Artemia nauplii.     

Brachionus vs Artemia duel: Optimizing first feeding of Upogebia pusilla (Decapoda: Thalassinidea) larvae

Larval rearing of many marine organisms is dependent on the availability of live food. The aim of this study was to optimize larval first feeding for the mud shrimp Upogebia pusilla, by comparing the effectiveness of the two most commonly used live feeds: Brachionus plicatilis and Artemia sp. nauplii. Survival, larval duration, molt synchronism and megalop size were compared using five feeding treatments: Artemia from zoea I to IV (B0), Brachionus during zoea I and Artemia from zoea II to IV (B1), Brachionus during zoea I and II and Artemia during zoea III and IV (B2), Brachionus from zoea I to III and Artemia during zoea IV (B3) and Brachionus from zoea I to IV (B4). The proportion of larvae that reached the megalop stage was 0.00% in treatment B0, 3.33% in treatment B1, 33.33% in treatment B2, 66.67% in treatment B3 and 76.67% in treatment B4. Larvae fed on rotifers until zoea III or zoea IV stages had a higher survival but no differences were found either in time to reach megalop or in megalop size. This study demonstrates that rotifers are essential for the survival and development of U. pusilla early larval stages but that rotifers can be successfully replaced by Artemia nauplii in the zoea IV stage.     

Enriching rotifers with “premium” microalgae. Isochrysis aff. galbana clone T-ISO

The effect of semi-continuous culture on the nutritional value of microalgae was tested in the rotifer Brachionus plicatilis in short-term enrichment experiments. Isochrysis aff. galbana clone T-ISO was cultured semi-continuously with renewal rates from 10 to 50% of the volume of the culture per day and used to feed the rotifers. After 24 h, dramatic differences in dry weight and protein, lipid and carbohydrate contents were observed in the rotifers depending on the renewal rate applied to the microalgal culture. Rotifers fed T-ISO cultured with low renewal rates showed low dry weight and organic content, whereas rotifers fed microalgae from nutrient-sufficient, high renewal rate cultures showed higher dry weight and increases up to 60% in protein, 35% in lipid and 100% in carbohydrate contents. Feed conversion rate (FCR) and organic FCR decreased with increasing renewal rates, indicating a more efficient assimilation of the microalgal biomass obtained at high growth rates. The fatty acid profile of rotifers reflected that of T-ISO, with maximum content of polyunsaturated fatty acids (PUFAs), n-3 fatty acids and docosahexaenoic acid (DHA) being found in the rotifers fed microalgae from the renewal rate of 40%. Results demonstrate that the biochemical composition of B. plicatilis is strongly modified through the use of semi-continuous cultures of microalgae in short-term enrichment processes. This technique provides an excellent tool to improve the nutritional value of the live feed used in fish larvae cultures.     

纳米硒对齐口裂腹鱼生长、肌肉成分、血清生化及抗氧化指标的影响

为探究饲喂富硒酵母强化卤虫无节幼体对大口黑鲈仔鱼的生长、存活率、脂肪酸组成、肝肠组织结构以及抗逆性的影响,设置了0 (Y-se0)、5 (Y-se5)、10 (Y-se10)、15 (Y-se15)、20 mg/L (Y-se20)酵母硒添加浓度对卤虫无节幼体进行硒营养强化,并使用强化后的卤虫饲喂养殖在玻璃缸中的3 000尾大口黑鲈仔鱼(2.1 mg/尾,200尾/缸) 20 d。饲养实验结束后,将每缸10尾仔鱼放入35 °C恒温水浴缸中进行急性温度胁迫试验。结果显示：① Y-se15组的存活率、终末体重、体长以及特定生长率均显著高于对照组,肥满度(CF)在各组间无显著差异。② Y-se5组EPA含量显著低于对照组,Y-se5及Y-se15组DHA/EPA值显著高于对照组。③ Y-se10、Y-se15和Y-se20组肝脏中过氧化氢酶(CAT)活性显著高于对照组,所有处理组肝脏中,谷胱甘肽过氧化物酶(GSH-Px)活性显著高于对照组,丙二醛(MDA)含量显著低于对照组,处理组和对照组间总超氧化物歧化酶(T-SOD)活性无显著差异。④ Y-se15组肝脏组织空泡化现象较轻,酵母硒添加组的肠道绒毛长度显著高于对照组。⑤ Y-se15组在35 °C应激条件下,存活时间显著高于对照组。研究表明,卤虫无节幼体强化液酵母硒添加浓度为15 mg/L时可显著提高大口黑鲈仔鱼生长、存活率及DHA/EPA水平,增强机体抗氧化以及抗应激能力,改善鱼体健康状况。本研究初步探究了饲喂酵母硒强化卤虫无节幼体对大口黑鲈仔鱼的影响,旨在为进一步完善水生动物苗种开口饵料营养强化技术提供参考,促进水产养殖业高质量发展。

     

Ontogeny of the digestive tract of thick lipped grey mullet (Chelon labrosus) larvae reared in “mesocosms”

This work describes the ontogeny of the digestive tract in thick lipped grey mullet (Chelon labrosus) larvae reared until day 36 post-hatching with the semi-extensive technology in mesocosms. Diet was constituted by live preys, rotifers, Artemia and wild zooplankton, then compound diet was added from day 20 (p. h.). Linear growth, weight growth and digestive enzymes specific activities were studied during larval ontogeny. Pancreatic enzymes (trypsin and amylase) and intestinal enzymes (leucine-alanine peptidase “Leu-ala”, aminopeptidase N “AN” and alkaline phosphatase “AP”) were assayed in larvae sampled throughout the rearing trial to evaluate gastrointestinal maturation along the development.The trypsin specific activities were very high during the first two weeks and then declined as observed in marine fish species. A following increase in trypsin specific activity from day 20 was attributed notably to ingestion of particle compound diet. In contrast to the pattern generally described in fish larvae, amylase specific activity showed a continuous increase. This could be attributed to the fact that C. labrosus is an omnivorous species and suggests that the fish might be able to use efficiently diets containing higher levels of starch or other carbohydrates since the end of larval development.Relative expression of intestinal brush border membrane enzymes (AP and AN) and cytosolic enzyme (Leu-ala), showed an abrupt increase of both AP/leu-ala and AN/leu-ala ratios at day 8 (p. h.), indicating that maturation of intestinal tract in C. labrosus larvae is particularly precocious. It is assumed that larvae of C. labrosus might support early co-feeding and weaning strategies, which could reasonably be initiated since mouth opening.     

Use of formulated diets as replacements for Artemia in the rearing of juvenile American lobsters (Homarus americanus)

American lobsters, Homarus americanus, have been successfully reared in hatchery operations for over a century, yet formulated diets have never been commercially produced. In recent years, commercial Artemia replacement (CAR) diets have been developed and marketed for use in aquaculture production of marine shrimp. Three separate experiments assessed the utility of rearing American lobsters on these shrimp CAR diets. First, survival and growth of stage IV American lobsters fed one of three CAR diets (Artemac 5, CAR1; Economac 4, CAR2; and Progression 3, CAR3) were compared to those of animals fed frozen adult n-3 fatty acid enriched Artemia. Survival to 3 months was highest for animals fed CAR3 (85%), while animals fed Artemia had the greatest weight gain (> 6 % day^− 1). A cost / benefit ratio analysis showed that CAR2 was the most cost efficient for juvenile production because of its low overall purchase cost. Second, stage IV lobsters were fed either CAR2 or frozen adult n-3 fatty acid enriched Artemia exclusively, or in combination (2 : 5, and 5 : 2). Again, CAR2 was a cost effective feed to use, even as a partial replacement for Artemia. Survival was higher in diets that included CAR2, and feeding it two days per week compensated for low quality Artemia. Finally, 1.5 year old lobsters fed a gelatin-bound mix of 80% CAR2 and 20% frozen Artemia for five months survived and grew equally well compared to lobsters fed gelatin-bound frozen adult Artemia, and better than a custom formulated maintenance diet. The benefits of incorporating formulated feeds into American lobster rearing programs to increase the effectiveness of enhancement programs is discussed.     

Beneficial effects of n-3HUFA enriched Artemia as food for larval palmetto bass (Morone saxatilis × M. chrysops)

The beneficial effects of feeding n−3 highly unsaturated fatty acids (HUFA ≥ 20 carbon fatty acids with three or more double bonds) to palmetto bass (striped bass × white bass) larvae, 4–30 days of age, were studied using Artemia diets enriched with six n-3HUFA levels. Dietary n-3HUFA concentrations were < 0.03% (control diet), 0.33%, 0.63%, 0.87%, 1.26%, and 2.27% of dry-wt Artemia. Larval n-3HUFA contents were reduced at a faster rate with decreasing dietary n-3HUFA intake, and were significantly different by 30 days posthatch (4–20 mg g⁻¹ dry-wt fish). Starved larvae selectively conserved endogenous n-3HUFA reserves, indicating an essential role of n-3HUFA in larval development. Mass mortality in the control and 0.33% n-3HUFA diets occurred at metamorphosis (26–28 days posthatch). At harvest, all fish, except those fed the two highest n-3HUFA diets, suffered from handling stress (shock syndrome) with increasing severity (25 to 100%) at decreasing dietary n-3HUFA intake. Recovery from shock syndrome was 100% at the 0.63% and 0.87% n-3HUFA diets, 63% at the 0.33% n-3HUFA diet and 0% at the control diet. Post-harvest survival was similar among the four highest dietary n-3HUFA groups (64–73%), whereas the two lowest n-3HUFA groups differed significantly (0 and 37%). Growth promotion was evident with increased dietary n-3HUFA intake as fish fed the highest n-3HUFA diet were twice the size of those fed the control diet (99 vs 52 mg wet-wt). Final fish sizes at the three highest n-3HUFA diets were similar. Given similar culture conditions, a minimum dietary n-3HUFA intake of 1.26% of dry-wt Artermia is recommended to avoid handling stress and promote growth in larval palmetto bass.     

“Indirect selection” for genetic change: A quantitative analysis illustrated with Macrobrachium rosenbergii

“Indirect selection” is exerted on a trait indirectly, by means other than artificial selection for the trait itself. Expressions are derived for calculating the intensity of indirect selection in aquaculture environments. The calculations are illustrated with growth-rate data on M. rosenbergii from farm ponds in Thailand, and with computer-generated data which simulate measurements made during a multiple mark-recapture experiment. Indirect selection for growth is probably negligible in Thai prawn farms and hatcheries at present, but small changes in management practice could exert strong indirect selection on growth rate. One very effective change appears to be collecting broodstock as early in the production cycle as possible. Control of indirect selection may be useful for the genetic improvement of aquaculture stock, especially in developing countries and other situations where an intensive artificial selection program is not economically or biologically desirable. Like all selection programs, the probability of success is critically dependent on the genetics of the traits being selected.     

Co-feeding of live feed and inert diet from first-feeding affects Artemia lipid digestibility and retention in Senegalese sole (Solea senegalensis) larvae

The present study intended to evaluate the effects of early introduction of inert diet in lipid digestibility and metabolism of sole, while larval feed intake, growth and survival were also monitored. Solea senegalensis larvae were reared on a standard live feed regime (ST) and co-feeding regime with inert diet (Art R). Trials using sole larvae fed with Artemia enriched with two different lipid emulsions, containing glycerol tri [1-¹⁴C] oleate (TAG) and L-3-phosphatidylcholine-1,2-di-[1-¹⁴C] oleoyl (PL), were performed at 9 and 17 days after hatching (DAH) to study lipid utilization. Co-feeding did not affect sole survival rates (ST 59.1 ± 15.9%; Art R 69.56 ± 9.3%), but was reflected in significantly smaller final weight at 16 DAH (ST 0.71 ± 0.20; Art R 0.48 ± 0.14 mg). Higher feed intake was observed in sole larvae fed on Artemia enriched with labeled PL at 9 DAH but not at 17 DAH. At 17 DAH, the smaller larvae (Art R treatment) ingested proportionally more Artemia in weight percentage, independently of enrichment. At 9 DAH lipid digestibility was equal among treatments and higher than 90%, while at 17 DAH it was higher in ST treatment (around 73%) compared to the Art R group (around 66%). Lipid retention efficiency at 9 DAH was higher in the Art R treatment, reaching values of 50%, while these values almost duplicated at 17 DAH, ranging up to 80% in both treatments without significant differences. These results show that co-feeding of live feed and inert diet from first-feeding in Senegalese sole has a toll in terms of growth and lipid digestibility but does not seem to compromise lipid metabolic utilization.     

Effects of varying Artemia enrichment on growth and survival of juvenile seahorses, Hippocampus abdominalis

Seahorses are the focus of recent aquaculture ventures in a variety of countries. Many of these ventures utilise enriched live Artemia (to varying degrees) to feed their seahorses. Given the range of commercially available enrichment products that can be used to enrich Artemia, it is imperative to determine which of these may promote the best growth and survival in seahorses, as well as being most cost-effective. This investigation tested the effects on juvenile seahorse (Hippocampus abdominalis Leeson, 1827) growth and survival through enriching Artemia with three commercial enrichment products: Super Selco®, DHA Protein Selco® and Algamac-3050. These were tested against a low-cost Artemia on-growing mixture used in this instance as an enrichment (90% Eyre Peninsula Aquaculture brine shrimp food (EPABSF)/10% spirulina).

After 3 months, there was a significant difference in juvenile length between the enrichment treatments, with juveniles in the DHA Protein Selco® and Algamac-3050 treatment longer than juveniles in the Super Selco® treatment, but not longer than juveniles in the EPABSF/spirulina treatment. There was also a significant difference in juvenile wet weights between the treatments, with juveniles in the Super Selco® treatment weighing significantly less than juveniles in the other three treatments.

In terms of condition factor (CF), there was a slight significant difference between the treatments, with juveniles in the DHA Protein Selco® and EPABSF/spirulina treatment having higher CFs than juveniles in the Super Selco® treatment, but not the Algamac-3050 treatment. Mean daily specific growth rate (SGR) for Super Selco® was lower than the other three treatments. There was no effect of enrichment treatment on seahorse survival with 100% survival in all four treatments.

Fatty acid analysis revealed marked differences in the Artemia enriched with the various enrichment products. Super Selco®- and Algamac-3050-enriched Artemia had the highest percentage of docosahexanoic acid (DHA), 22:6(n−3), while Super Selco®-enriched Artemia had the highest percentage of eicosapentaenoic acid (EPA), 20:5(n−3). EPABSF/spirulina-enriched Artemia had the highest percentages of C18 fatty acids. Percentage of polyunsaturated fatty acids (PUFA) and n−3 PUFAs were highest in Super Selco®- and Algamac-3050-enriched Artemia. Proximate analysis revealed little difference among all four enrichment treatments in terms of protein, fat and carbohydrate levels.

The results from this experiment demonstrate that when culturing H. abdominalis, all three commercial enrichment products produce good seahorse growth and excellent survival. However, on a cost/benefit basis, all three commercial products were outperformed by the cheaper Artemia on-growing mixture (EPABSF/spirulina).     

Comparison of growth and survival of white shrimp postlarvae (Litopenaeus vannamei) fed dried Artemia biomass versus four commercial feeds and three crustacean meals

The use of dried Artemia biomass meal as an exclusive feed for postlarval white shrimp (Litopenaeus vannamei) was compared with four commercial feeds and three crustacean meals in a series of trials. Postlarvae (PL1–PL6) were stocked at a density of 1.5–2.5/litres in 16 tanks (100 litres volume) and fed, ad libidum, five times a day, over 23–29 days. Feeding postlarval shrimp with dried Artemia biomass resulted in a significantly larger size than feeding with three of the commercial feeds, and the crustacean meals. There was no significant size difference observed in animals fed with Artemia biomass and the commercial 'Golden Pearls' feed for postlarvae, however the coefficient of variation among the size of the 'Golden Pearls' fed animals was significantly higher. The weight increase of animals fed with Artemia biomass was higher than in animals fed with all the tested feeds. The survival rate was not significantly different in animals fed with Artemia flakes from 'Salt Creek', 'Bio-Marine', 'Golden Pearls' and Artemia biomass, however the survival rate was significantly larger in animals fed with Artemia biomass than in animals fed with the crustacean meals and 'Artemac.' Results suggest that dried Artemia biomass is a well-suited feed for postlarval L. vannamei.     

Effect of docosahexaenoic acid enrichment in Artemia on growth of Pacific bluefin tuna Thunnus orientalis larvae

Feeding enriched Artemia induces growth failure in Pacific bluefin tuna (PBT) Thunnus orientalis larvae; however, feeding of yolk-sac larvae of marine fish promotes larval growth. It is considered that this growth failure partly results from dietary docosahexaenoic acid (22:6n-3, DHA) deficiency. Therefore, we examined the effect of DHA contents in enriched Artemia on the growth of PBT larvae. Artemia nauplii were enriched with graded levels of DHA ethyl ester, and fed to PBT larvae for 9 days. Yolk-sac larvae of Japanese parrotfish Oplegnathus fasciatus were used as a reference diet. The DHA contents in Artemia increased with the enrichment from 0 mg g^− 1 dry weight basis (DW) to 25 mg g^− 1 DW, while the content in the reference diet was 21 mg g^− 1 DW. Feeding of enriched Artemia significantly improved the growth of PBT larvae. However, this improvement was negligible when compared with the excellent growth of the larvae that were fed the reference diet. PBT larvae that were fed the reference diet accumulated approximately twice or much higher levels of DHA in the neutral and polar lipids in the body when compared with the larvae that were fed enriched Artemia. These results show that PBT larval growth cannot be promoted by feeding enriched Artemia even if the DHA contents in Artemia are elevated to the same levels as those of yolk-sac larvae. The incorporation of dietary DHA into phospholipids in the fish body may be desirable for the normal growth of PBT larvae.     

Rapid differentiation of Artemia spp. Populations by thin layer isoelectric focusing

Artemia spp. of various geographic origins, yearly collection lots, developmental stages, and a binary geographic mixture were analyzed by thin layer polyacrylamide gel isoelectric focusing (IEF). IEF of geographical Artemia samples yielded unique profiles for each population in the pH range employed. Parthenogenetic populations of Artemia exhibited IEF patterns which were, generally, dominated by fewer proteins; bisexual populations exhibited more evenly distributed, complex protein patterns. There were similarities reflected in IEF profiles of geographically isolated Artemia descended from a common ancestry. Yearly collections of Artemia exhibited no apparent differences in IEF protein patterns. There were differences noted in three developmental stages of Artemia. A binary mixture of geographic populations yielded IEF patterns intermediate to those of the component pure samples.     

Inter-specific variation of the mitochondrial r16S gene among silversides, “Peces Blancos”, (Atherinopsidae: Menidiinae) and its utilization for species identification

Analysis of 570 nucleotide sequences of the mitochondrial r16S gene of four species of silversides “peces blancos” (Chirostoma sp.) and of two euryhaline related species Membras martinica and Menidia menidia indicated variation that aided in development of a DNA-based method for species identification. The nucleotide sequence data showed levels of inter-generic genetic variation that ranged from 3.3–7.9% and from 1.06 to 1.6% at the inter-specific level. The variation observed allowed the identification of 18 diagnostic restriction enzymes that in any combination of two or three could aid in the diagnosis of species of “peces blancos”; six of these enzymes could also discriminate between the genera Membras and Menidia; the enzymes AseI, CviAII and FatI discriminate the genus Membras from Chirostoma; and the combination of any of the enzymes BsaAI, CviAII, FatI or HpyCH4IV discriminate Menidia from the “peces blancos”. PCR-RFLP analysis of amplified mitochondrial r16S fragments of 90 un-identified “peces blancos” from Lake Patzcuaro utilizing the restriction enzymes HpyCH41V, TaqI, BsmF1 and BbvI allowed us to diagnose unambiguously cultivated Chirostoma estor and C. humboldtianum in juvenile and adult stages. The use of a non-invasive method to PCR-amplify specific DNA markers from live brood stocks suggests use of this tool for identification of endangered or threatened species.     

Domestication affects spawning of the ide (Leuciscus idus)—preliminary study

Ide, Leuciscus idus, breeders in Polish fish farm are cultured or are collected from the wild. The data about differences in reproduction of different ide forms in captivity are limited. In the present paper domesticated and wild stock are reproduced artificially. Cultured fish do not require application of spawning agents. For obtaining good-quality oocytes only stimulation with temperature and photoperiod is necessary.