Volume 64, 2012

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Genetic Diversity among Wild and Cultured Stocks of Sparus aurata on Turkish Mediterranean Coasts Revealed by Mitochondrial DNA Sequences
(2012) Dognakaya, L.; Bekcan, S.
The sequences of three mitochondrial DNA genes - 12S rRNA, cytochrome b, cytochrome oxidase II - were used to deduce genetic diversity and relationships among wild and cultured populations of gilthead sea bream (Sparus aurata) on Turkish coasts and coastal waters. The 415 bp of 12S rRNA, 605 bp of cytochrome oxidase II, and 557 bp of cytochrome b genes were sequenced and the data set was analyzed with Neighbor Joining and Minimum Evolution. Bootstrap analyses (1000 replicates) were performed and the relationships between samples are given in phylogenetic trees. Genetic diversity was estimated using gene diversity, number of haplotypes, and nucleotide diversity. There were no significant differences in variation in any of the three gene sequences. The maximum genetic variation occurred in cytochrome b (0.0050). Similarly, results showed very little divergence between sampling sites. Findings indicate a single wild population in the eastern Mediterranean, supporting the current classification that lacks subdivisions. Further, no differentiation between wild and culture stocks was inferred.
Nibbling Frequency of Carps in Periphyton-Based Aquaculture Systems with and without Supplemental Feed
(2012) Rai, Sunila; Yi, Yang
The nibbling frequency of five carp species (rohu Labeo rohita, mrigal Cirrhinus mrigala, catla Catla catla, common carp Cyprinus carpio, silver carp Hypophthalmichthys molitrix) on bamboo lateral sticks (kanchi) colonized by periphyton was examined in fed and unfed systems. There were three treatments: (a) no carp and no supplemental feed (control), (b) carp without supplemental feeding (unfed treatment), and (c) carp with supplemental feeding (fed treatment). For 12 h (07:30-19:30) during six days, nibbling behavior was observed in real time via a digital video camera and recorded on a camcorder for later viewing. Rohu, catla, and common carp nibbled on the kanchi, while mrigal and silver carp did not. In rohu and catla, the nibbling frequency was significantly higher in the unfed treatment than in the fed treatment (p<0.05); supplemental feeding reduced nibbling frequency by 81% and 91%, respectively. Hence, in periphyton-based aquaculture systems, there is no need for a high density of substrates in ponds that receive supplemental feed. Alternatively, the amount of supplied feed can be reduced to force these species to consume more periphyton.
Effects of Dietary Fish Oil Substitution with Palm Oil on Growth, Survival, and Muscle Proximate Composition of Cirrhinus mrigala (Hamilton, 1822)
(2012) Singh, Soibam Khogen; Rather, Mohd Ashraf; Mandal, Sagar C.; Das, Pronob; Pawar, Nilesh; Singh, Y. Jackie; Dar, Shabir Ahmad
A 60-day growth experiment was carried out to evaluate the effect of replacing dietary cod liver oil with palm oil on the growth, survival, and muscle proximate composition of the Indian major carp, Cirrhinus mrigala. Five diets were fed to triplicate groups of 20 advanced C. mrigala fry (2.16±0.01 g): 100% cod liver+0% palm (control), 25% palm+75% cod liver, 50% palm+50% cod liver, 75% palm+25% cod liver, and 100% palm+0% cod liver. At the end of the experiment, mean weight gain, average daily growth, specific growth rate, feed conversion efficiency, feed conversion ratio, and percentage survival were assessed. Growth performance and survival were not compromised when fish oil was substituted with palm oil up to 25%. However, beyond this level, growth was significantly retarded (p<0.01). There were no significant (p>0.01) differences in feed efficiency or muscle proximate composition. Results confirm the feasibility of substituting up to 25% dietary fish oil by palm oil in diets for C. mrigala advanced fry without negatively affecting growth or feed utilization efficiency.
Effect of Stocking Density on Growth and Survival of Sub-Adult Tench (Tinca tinca Linnaeus 1758)
(2012) Pantazis, Panagiotis A.
Three hundred and fifty-nine (359) sub-adult tench were allocated to twelve tanks to investigate the effect of stocking density on survival and growth. The experimental population consisted of two weight groups (small = 11.27-11.36 g and large = 15.38-15.44 g) and two initial stocking densities (1.4 kg/m3 and 2.5 kg/m3). The experiment lasted seven months (217 days). The total lack of deformities in the caudal peduncle in all groups and the low incidence of mortality indicate that sub-adult tench cultured in a water recirculation system perform well when stocked at a relatively high stocking density (2.5 kg/m3) and fed 1.0% of their body weight per day. By the end of the experiment, the weight of the best-performing groups increased 83-90% and confirmed the hypothesis that high densities favor the growth and survival of sub-adult tench reared in artificial tanks in a water recirculation system.
An Improved Method of DNA Extraction from the Shell of the Pacific Oyster, Crassostrea gigas
(2012) Zhang, Guofan; Wang, Xiaotong; Song, Xiaorui; Li, Li
We earlier developed a method to extract DNA from in vivo-sampled mantle tissue of the bivalve Pacific oyster (Crassostrea gigas) for molecular marker-assisted selection (MAS) in breeding. However, mortality was too high when using this method. In the current study, a more efficient and safer method of extracting DNA from the oyster shell was developed, based on the improved phenol-chloroform DNA extraction method. Both nuclear DNA and mitochondrial DNA were successfully extracted from the oyster shell, and mortality was 0%. These results illustrate the safety of extracting DNA from the bivalve shell. In addition, because DNA is a cellular component, the results provide molecular evidence that cells are likely involved in shell formation.
Sperm Characteristics of Wild European Flounder (Platichthys flesus luscus)
(2012) Sahin, Temel; Günes, Erdinç; Aydin, Ilhan; Kurtoglu, Ilker Zeki
The spermatologic characteristics of European flounder (Platichthys flesus luscus) were determined. Flounder were collected during the spawning season and sperm of eight males was extracted by abdominal massage. Body weight and total length of the flounder were measured, volume, motility, duration of motility, spermatocrit, density, and pH of the sperm were determined, and correlations between the spermatologic characteristics and weight and length were investigated. Mean values were 0.7±0.16 ml for sperm volume, 87.5±3.66% for motility, 22.0±1.49 min for duration of motility, 94.0±1.22% for spermatocrit, 2.7±0.16 × 109/ml for density, and 6.9±0.05 for pH. Body length and sperm volume had positive correlations with body weight (p<0.01), but the correlation between length and sperm volume was negative (p<0.01). Likewise, the correlations between spermatocrit and total length, sperm volume, and density were negative (p<0.05).
Pigmentary and Zootechnical Responses of Juvenile Litopenaeus vannamei (Boone, 1931) Maintained on Diets Supplemented with Xanthophylls of Marigold Tagetes erecta Flowers
(2012) Piña-Valdéz, Pablo; Aguirre-Hinojosa, Eduardo; Garza-Aguirre, María del Carmen; Montoya-Olvera, Ricardo; Torres-Quiroga, José Odón; Nieves-Soto, Mario
Practical diets supplemented with 75 or 150 ppm xanthophylls (75% zeaxanthin, 15% lutein) industrially extracted from marigold (Tagetes erecta L.) flowers increased the astaxanthin and total carotenoid concentrations in juvenile Litopenaeus vannamei, compared to shrimp fed a practical control diet. Our results paralleled or exceeded those obtained with a diet containing 75 ppm supplementary synthetic astaxanthin. The post-feeding astaxanthin concentration accounted for more than 84% of the total carotenoid concentration in shrimp fed either diet, while beta-carotene, zeaxanthin, lutein, and other non-identifiable carotenoids comprised a minority of the total concentration. That this was seen in both the tail exoskeleton and abdominal muscle indicates that L. vannamei can metabolize precursor xanthophylls to produce astaxanthin. In most cases, more than 60% of the astaxanthin was esterified. In general, survival improved in shrimps fed the supplemented diets compared to those fed the control diet. There were no differences in growth.
Stress Survival in Larvae of Florida Pompano (Trachinotus carolinus) Fed Enriched Rotifers (Brachionus plicatilis) and Nauplii of the Calanoid Copepod (Pseudodiaptomus pelagicus)
(2012) Cassiano, Eric J.; Ohs, Cortney L.; DiMaggio, Matthew A.
The Florida pompano, Trachinotus carolinus, is a highly prized marine fish whose larviculture includes the feeding of live rotifers and brine shrimp nauplii. In a previous study, growth and survival of pompano larvae fed nauplii of the calanoid copepod, Pseudodiaptomus pelagicus, were compared to those of larvae fed enriched rotifers, Brachionus plicatilis. There were advantages to including the copepod in the larvae diet. The current study examines the stress tolerance of such larvae. Two trials were conducted: for seven (trial 1) and nine (trial 2) days post-hatch. Larvae were fed diets that included enriched rotifers and/or P. pelagicus nauplii and subjected to varying durations of air exposure (‘sieve stress’). Larvae fed copepods exhibited significantly greater stress tolerance than larvae fed only enriched rotifers. In trial 1, stress tolerance increased as the number of days on which copepods were fed increased. It is possible that stress tolerance improved because of a better nutritional profile of the copepod nauplii.
Effects of Total Replacement of Fishmeal with Spirulina Powder and Soybean Meal on Juvenile Rainbow Trout (Oncorhynchus mykiss Walbaum)
(2012) Hernández, Luis Héctor Hernández; Flores, Gerardo Hernández; Araiza, Mario Alfredo Fernández; López, Omar Angeles
Juvenile rainbow trout (Oncorhynchus mykiss) were fed diets in which fishmeal was totally replaced by mixtures of Spirulina powder and soybean meal. Three experimental diets were formulated with 75% Spirulina and 25% soybean meal, 50% Spirulina and 50% soybean meal, and 25% Spirulina and 75% soybean meal. A commercial diet and a diet with 100% fishmeal served as controls. Diets were fed to triplicate groups of 10 juveniles with an initial weight of 6.8±0.4 g for 50 days. At the end of the experiment, growth performance, oxygen consumption, nitrogen and phosphorus excretion, protein digestibility, and serum protein content and lysozyme activity were evaluated. Data were analyzed by one-way ANOVA. Growth performance was not significantly affected by the experimental diets, but values tended to be lower as the level of dietary soybean meal increased. Among the experimental diets, the 75% Spirulina/25% soybean diet produced the best growth performance. Further, in fish fed this diet, the apparent coefficient of digestibility of protein was higher than 98% (as with all diets), oxygen consumption was lower and lysozyme activity was higher than in fish fed other replacement diets, and dissolved phosphorus and phosphorus in feces were similar or lower than in the controls. The present research shows the possibility of using mixtures of Spirulina powder and soybean meal as total substitutes of fishmeal in diets for juvenile rainbow trout.
Effect of Dietary Supplementation of Achyranthes aspera Seed on the Immune System of Labeo rohita Fry
(2012) Chakrabarti, R.; Srivastava, P. Kumar
Labeo rohita (rohu) fry (0.547±0.01 g) were fed one of four diets containing 0 (control), 0.1, 0.5, or 1.0% Achyranthes aspera seed. After 30 days, the fry were immunized with chicken red blood cells (c-RBC). Blood samples were collected 7, 14, and 21 days after immunization. Significantly (p<0.05) higher average weight (2.565±0.02 g) and SGR were were obtained in fry fed the 1.0% diet compared to others. The increase in average weight was directly related to the increasing dose of seed. FCR was significantly (p<0.05) lower in fry fed the 1.0% diet while total serum protein, albumin, and globulin were higher in treated groups than in the unsupplemented control. The antigen-specific antibody titer level was significantly (p<0.05) lower in rohu fed the control diet than in rohu fed the supplemented diets. Titer levels were 32-128, 128-256, 256-1024, and 256-1024 in the 0, 0.1, 0.5, and 1.0% diets, respectively. Serum glutamic oxaloacetic transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), and alkaline phosphatase (ALP) were significantly (p<0.05) higher in rohu fed the control diet than in rohu fed the supplemented diets while myeloperoxidas was lower in the former. Among the treated groups, myeloperoxidase was significantly (p<0.05) higher in rohu fed the 1.0% diet (1.561-2.558, ? 450 nm) than in those fed other diets. The present study documents the immunostimulatory properties of A. aspera seed and finds that a dose of 1.0% might be suitable for rohu fry.
Low Mercury Levels in Lake Kinneret Fish
(2012) Blanchfield, Paul J.; Shapiro, James; Sukenik, Assaf; Orihel, Diane M.; Shefer, Edna
Human exposure to the neurotoxin, methylmercury (MeHg), occurs primarily through the consumption of contaminated fish that are present even in pristine freshwater ecosystems. Lake Kinneret is the sole freshwater lake in Israel and supports an important commercial fishery. We examined total mercury (THg; often equivalent to MeHg in fish muscle tissue) for nine fish species in Lake Kinneret. Concentrations were low for all fish species, 6-409 ng/g, and below 500 ng/g MeHg, the level generally considered safe for human consumption. Of particular relevance are the very low levels of THg (<60 ng/g) in Galilee St. Peter’s fish (Sarotherodon galilaeus) and lavnun (Acanthobrama terrascantae), the most commercially important species in Lake Kinneret. Biomagnification was apparent among trophic guilds, with THg concentrations increasing from primary to secondary to tertiary consumers. This study suggests that consumption of commercial species, especially primary consumers, will result in low MeHg exposure to humans.
Level of L-ascorbyl-2-monophosphate-Mg as a Vitamin C Source in Practical Diets for the Asian sea bass, Lates calcarifer
(2012) Catacutan, Mae R.; Pagador, Gregoria E.; Doyola-Solis, Ellen Flor; Ishikawa, Manabu; Teshima, Shinichi
The stable vitamin C derivative, L-ascorbyl-2-monophosphate-Mg (AMP), was used as a supplement in practical diets for Asian sea bass, Lates calcarifer. Optimum growth, liver ascorbic acid (AA) saturation, and wound healing were determined. Sea bass (78.9±0.4 g) were fed a maintenance diet without vitamin C supplement for 25 days, then distributed into fifteen 1500-l oval fiberglass tanks at 30 fish each and fed one of five practical diets containing 0, 50, 100, 200, or 400 AMP mg/kg diet for 14 weeks. Fish fed the AMP-free diet exhibited clinical signs of vitamin C deficiency and significantly poorer final average weight, specific growth rate, protein efficiency ratio, feed conversion ratio, and hematocrit level (p<0.05). The level of AA in the sea bass brain increased as the level of dietary AMP increased. The AA concentration in the liver was similar at all AMP dietary levels. Body calcium of sea bass fed the AMP-free or 50 mg diets was significantly lower (p<0.05) than in fish fed the other diets. AMP enhanced wound healing regardless of supplement level, but fish fed the highest dietary AMP (400 mg/kg diet) exhibited histopathological changes in the liver. The dietary level of 50 mg AMP/kg diet was adequate for optimum growth, liver AA saturation, and prevention of clinical signs of vitamin C deficiency in 80-220 g sea bass. However, 100 mg AMP per kg diet was necessary for optimum body calcium.
Zinc Supplementation and its Effect on Thermal Stress Resistance in Carassius auratus Fry
(2012) Babitha Rani, A. Hasnat; Kohli, M.P.S.; Chandraprakash, G.
Goldfish (Carassius auratus) fry (80 mg) were fed purified diets based on casein as a protein source and containing different levels of supplementary zinc (0, 30, 60, 90, 120, 150 mg Zn/kg diet) for nine weeks. There were overt signs of zinc deficiency (suppressed growth, high mortality) in fish fed the zinc-deficient control diet, while the highest growth was obtained in fish fed the 60 mg Zn/kg diet. Alkaline phosphatase (ALP) activity in the muscle and super-oxide dismutase (SOD) activity in the liver were highest in fish fed the 60 mg Zn/kg diet and did not significantly improve beyond this level. Muscle and whole body zinc concentrations increased proportionately with the dietary zinc supplementation but iron and copper concentrations in fish tissue were not affected by the supplementary zinc. After the 9-week trial, twelve fish were subjected to thermal stress (28-32°C) for another four weeks to observe growth and the physiological response of the stress enzyme, SOD. After thermal stress, growth improved with the increase in zinc. Likewise, SOD activity in the fish liver increased, indicating that fish utilize more zinc in high temperatures to counteract stress.
Effects of Dietary Pyridoxine on Growth and Physiological Responses of Labeo rohita Fingerlings Reared in High Water Temperature
(2012) Akhtar, M.S.; Pal, A.K.; Sahu, N.P.; Ciji, Alexander; Meena, D.K.
The role of dietary pyridoxine (vitamin B6) on the growth and physiological responses of Labeo rohita fingerlings reared in a high temperature of 33°C was examined in a 60-day trial. Two hundred and seventy fingerlings (6.71±0.32 g) were randomly distributed into six treatments in triplicate. Five iso-caloric and iso-nitrogenous purified diets were prepared with graded levels of pyridoxine: 0, 10, 50, 100, and 200 mg. The control group was given a diet containing 10 mg pyridoxine and reared at ambient temperature (26°C). Pyridoxine supplementation at 100 or 200 mg/kg diet significantly augmented the specific growth rate but there were no significant (p>0.05) effects of pyridoxine supplementation on the feed conversion ratio. Superoxide-dismutase, catalase, blood glucose, and serum cortisol were significantly lower and acetylcholinesterase (AchE) significantly (p<0.05) higher in the pyridoxine-fed groups. Results indicate that dietary pyridoxine supplementation at the rate of 100 mg/kg diet augments growth and helps lower stress in L. rohita fingerlings reared in a high water temperature.
Detection of Acylated Homoserine Lactone (AHL) in the Heterotrophic Bacteria Z-TG01 and Its Ecological Action on the Algae, Chlorella vulgaris
(2012) Tang, Xuexi; Bi, Xiangdong; Xing, Kezhi; Zhou, Wenli
Heterotrophic bacteria Z-TG01 was isolated from the phycosphere of Chlorella vulgaris and identified as Sphingomonas paucimobilis by VITEK 2 Compact automated system with an excellent confidence level (99.0% probability). We detected acylated homoserine lactones (AHL) bioactivity in the growth process of Z-TG01 using Agrobacterium tumefaciens KYC55. The effects of C. vulgaris on AHL bioactivity, as well as the mutual influence of C. vulgaris and Z-TG01 in a co-culture system, were investigated. Results showed that AHL bioactivity of bacteria Z-TG01 varied with the bacteria density and was inhibited by supernatants of C. vulgaris culture in the early and middle exponential phases within 2 h (p<0.01), and in the late exponential phase within 1.5 h (p<0.05 or p<0.01). Within a certain cell density, bacteria Z-TG01 and C. vulgaris promote mutual growth in a co-culture system.
Ultrastructure of Erythrophores and Xanthophores of the Siamese Fighting Fish, Betta splendens
(2012) Khoo, Gideon; Lim, Tit Meng; Phang, Violet P.E.
The ultrastructural morphology of brightly colored pigment cells (chromatophores) of Siamese fighting fish, Betta splendens, was investigated by transmission electron microscopy. The major pigment cells in the epidermis and dermis of the red and golden strains were erythrophores and xanthophores, respectively. Specific combinations of these chromatophores formed the basis of pigmentation patterns in both strains. The ultrastructure of the erythrophores was characterized by ellipsoidal electron-lucent vesicles that had limiting membranes and inner lamellae. The latter appeared whorl-like due to a concentric arrangement of parallel membranes. The xanthophores contained small and large cytoplasmic vesicles that appeared hollow and electron-lucent, with some vesicles displaying slightly electron dense particles. Sections of some large vesicles also revealed a very thin membrane enveloping these droplet-like vesicles.
Polysaccharides, Saponins, and Water Decoction of Astragalus membranaceus Significantly Enhance the Non-Specific Immune Response of Spotted Maigre (Nibea albiflora)
(2012) Wang, Xuepeng; Ding, Lei; Yan, Maocang; Chai, Xueliang; Lu, Rongmao; Wang, Qishuo; Li, Fuchang
The effects of polysaccharides, saponins, and water decoction of the Chinese herb Astragalus membranaceus on the immune response of spotted maigre (Nibea albiflora) were investigated. Fish with an average initial weight of 49.6±5.5 g were fed a diet containing 2% extract for four weeks. The lysozyme and phagocytic activities of the fish were determined and compared with those of control fish fed an unsupplemented feed on days 0, 3, 7, 14, 21, and 28. Both activities were significantly higher in groups fed the Astragalus extract than in the control group (p<0.01) but there were no differences between the three Astragalus-fed groups (p>0.05). Most important, the Astragalus-fed groups were significantly protected against Vibrio vulnificus challenge compared to the control group (p<0.01). Our results indicatd that polysaccharides and saponins are main active components of Astragalus extracts and can enhance the non-specific immune response in spotted maigre.
Growth and Survival of Anadara inaequivalvis (Bruguiere, 1789) in Sufa Lagoon, Izmir, Turkey
(2012) Acarli, Sefa; Lok, Aynur; Yigitkurt, Selcuk
The growth and survival of 5, 7, 9, and 11-mm juvenile blood cockle (Anadara inaequivalvis) cultured in suspended nets in Sufa Lagoon, Izmir, Turkey, were investigated. Shell length, width, and thickness and total weight were measured monthly during May 2007-May 2008. Temperature was measured every 6 h. Salinity, chlorophyll a, particulate organic matter, particulate inorganic matter, and total particulate matter were followed every two weeks. Mean increases were 16.68 and 13.46 mm and 3.62 and 5.74 g for small and large spat, respectively. Length increased significantly faster in small cockle than in large cockle (p<0.05). Survival was 65% and 100% in small and large cockles, respectively (p<0.05). Von Bertalanffy growth parameters L∞ (27 mm) and k (0.28/month) showed that growth performance (Ф′ = 3.22) was greater than for other species in the natural population. Slope b from the length-weight relationship was 3.098±0.115, indicating significantly better allometric growth (p≤0.05). Results indicate that A. inaequivalvis is a good candidate for suspended aquaculture in the Sufa Lagoon, Turkey.
Effect of Marigold Flower and Beetroot Meals on Growth Performance, Carcass Composition, and Total Carotenoids of Snow Trout (Schizothorax richardsonii)
(2012) Jha, Ghanshyam Nath; Sarma, Debajit; Qureshi, T.A.; Akhtar, M.S.
A 60-day experiment was carried out to elucidate the effect of marigold (Tagetes erecta) flower and beetroot (Beta vulgaris) meals on growth performance, carcass composition, and total carotenoids of snow trout, Schizothorax richardsonii. Two hundred and seventy fingerlings (9.19±0.29 g) were randomly distributed into nine treatments in triplicate (10 fish per tank). Nine isonitrogenous (35.25±0.9% crude protein) diets were prepared with graded levels (3%, 5%, 7%, 10%) of either marigold flower meal or beetroot meal; the control diet contained neither marigold flower nor beetroot meal. Weight gain and specific growth rate were significantly (p<0.05) higher in fish fed the diet containing 10% beetroot meal than in those fed the control diet. Body carotenoid was significantly enhanced (p<0.05) by the dietary supplements and increased linearly with the increase of marigold flower meal (Y = 0.532x + 1.126, R2 = 0.9803) and beetroot meal (Y = 0.491x + 1.341, R2 = 0.9376). Results indicate that inexpensive and readily available natural carotenoid sources such as marigold flower and beetroot meals can be incorporated into diets for S. richardsonii to enhance pigmentation and ornamental value.
Effect of Astacin on Growth and Color Formation of Juvenile Red-White Ornamental Carp (Cyprinus carpio var. koi L)
(2012) Liang, Yong-jun; Bai, Dong-qing; Yang, Guang; Wei, Dong; Guo, Mei; Yan, Shan-shan; Wu, Xuan; Ning, Bo
Body color is an important quality and determines the value of koi. Fish are unable to fully synthesize carotenoid pigments. Therefore, carotenoid needs to be added to their diets. This study determined the effect of astacin on the growth and body color enhancement in healthy red-white ornamental carp (Cyprinus carpio var. koi). Fish were fed diets containing 0 (control), 50, 100, 150, 200, or 250 mg astacin/kg diet for 8 weeks. At the end of the experiment, samples of red-white, red, and white skin, head, scales, and fin ray were collected for carotenoid content analysis and visual observation. Fish weight and specific growth rate were significantly higher in fish fed the 150, 200, and 250 mg diets than in fish fed the control while the feed conversion rates in the 200 and 250 groups were significantly lower than in the control. Carotenoid deposition in the skin, head, scale, and fin ray of fish fed diets containing astacin was significantly higher than that those fed the control. In addition, a higher astacin content resulted in a brighter body color. The highest carotenoid concentration was achieved when 250 mg astacin/kg feed was added. Astacin was mainly deposited in the skin, scales, and head.

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