Evaluation of three seaweeds Gracilaria bursa-pastoris, Ulva rigida and Gracilaria cornea as dietary ingredients in European sea bass (Dicentrarchus labrax) juveniles

The aim of this study was to evaluate the inclusion of three seaweeds Gracilaria bursa-pastoris (GP), Ulva rigida (UR) and Gracilaria cornea (GC) as dietary ingredients on the performance, nutrient utilisation and body composition of European sea bass juveniles. Six experimental diets were formulated to replace 5% (GP-5, UR-5, and GC-5 Diets) and 10% (GP-10, UR-10 and GC-10 Diets) fish protein hydrolysate (CPSP) by each of the three seaweeds. A control diet was used, without inclusion of any seaweed. Diets were fed to duplicate groups of 25 European sea bass (Dicentrarchus labrax) juveniles (IBW = 4.7 g) for 10 weeks. Growth performance was only significantly reduced (P < 0.05) in fish fed the GC-10 diet, whereas the feed conversion ratio increased significantly in those fish. The apparent digestibility coefficients of dry matter and lipid were significantly lower in fish fed diet GC-10 relative to those fed the control diet. Carcass composition was similar among treatments, although fish fed GC-10 exhibited significantly higher ash content.The results obtained in this study suggest that the inclusion of G. bursa-pastoris (GP) and U. rigida (UR), up to 10%, can be considered as very interesting ingredients in diets for sea bass juveniles, as no negative consequences on growth performance, nutrient utilization or body composition were observed. On the other hand, the inclusion of G. cornea (GC) should be limited to 5% of the diet.     

Identification and characterization of a green-type mutant of Porphyra tenera Kjellman var. tamatsuensis Miura (Bangiales, Rhodophyta)

We identified two green-colored conchocelis strains of cultivated Porphyra, ‘Oba-green’ and HGT-6 (F₁ strain of ‘Oba-green’), based on PCR-RFLP and sequence analyses of the nuclear ITS region and the plastid RuBisCO spacer region. These molecular analyses confirmed that the two strains were green-type mutants of the endangered species Porphyra tenera, although their blade color was not green. Under the same culture conditions, blade length increase was significantly higher in HGT-6 than in a wild strain HGT-1 of P. tenera, and the blade shape of HGT-6 was extremely elongate compared with that of HGT-1. From the growth characteristics and ITS-1 sequence data, HGT-6 was confirmed to be a green-type mutant of P. tenera var. tamatsuensis, a vigorously growing cultivar. Although photosynthetic pigment contents were lower in HGT-6 than in HGT-1, total content of four major free amino acids was higher and the blade thickness was almost the same in HGT-6 and HGT-1. These results suggest that the green-type mutant HGT-6 has potential as breeding material for further development of Porphyra breeding.     

The mariculture potential of the Indonesian reef-dwelling sponge Callyspongia (Euplacella) biru: Growth, survival and bioactive compounds

In the present study, the Indonesian reef-dwelling sponge Callyspongia (Euplacella) biru de Voogd [de Voogd, N.J., 2004. Callyspongia (Euplacella) biru spec. nov. (Porifera: Demospongiae: Haplosclerida) from Indonesia. Zool. Meded. 78, 477-483.] (Demospongiae, Callyspongiidae) was cultured for a period of 6 months at three sites in the Spermonde Archipelago, Indonesia. A total of 250 sponge cuttings of two size classes (30 and 40 mm in length) were threaded on either polyethylene or very fine nylon rope and attached to horizontal mooring systems. All sponge cuttings were photographed at regular time intervals. Survival rates were high (82%-100%) among all treatments used. Growth rates differed significantly among treatments, but did not differ significantly per location, farming method, explant size and explant position obtained from the parent sponge. The concentration of the bioactive compound amphitoxin in the cultured explants was not significantly lower than in the natural population, but the concentration did vary significantly between explants used in the different treatments, and also between the different fragments of the natural population. The high growth potential and high survival rate suggest that this sponge species is a promising candidate for further mariculture development.     

Some Aspects of the Growth of Gracilaria birdiae (Gracilariales, Rhodophyta) in an Estuary in Northeast Brazil

The agarophyte Gracilaria birdiae Plastino & Oliveira was cultivated under field conditions in an estuary over a 6-month period. Biomass production varied significantly (p<0.01) and ranged from 900.0 to 3537.0 g m⁻² with a mean of 2124.4±1004.36 g m⁻² over the cultivation period. The epiphyte-biomass varied from 64.7 to 313 g m⁻², with maximum values observed in July. Relative growth rate (RGR) of G. birdiae varying from −0.59 to 4.67% day⁻¹, with a maximum mean observed in July. Correlation analysis showed that RGR was positively correlated with epiphyte-biomass (R-Pearson=0.69; p< 0.01) and negatively correlated with salinity (R-Pearson=−0.41; p<0.05). These correlations were used in a mathematical regression model to estimate the growth of Gracilaria in an estuary. The regression model explained 61% of RGR variability demonstrating that growth rates of Gracilaria were directly related to the variables salinity and epiphytes. The model was developed to help to predict production at a cultivation site in order to evaluate its suitability for Gracilaria cultivation.     

Heritability for body weight at harvest size in the Pacific white shrimp, Penaeus (Litopenaeus) vannamei, from a multi-environment experiment using univariate and multivariate animal models

To estimate family BLUP breeding values and the heritability of body weight at harvest size (BW) in the Pacific white shrimp, Penaeus (Litopenaeus) vannamei, an experiment was conduced using information from two farm units of a Mexican hatchery and two shrimp population densities at each location. Data consisted of 12,658 shrimps that were siblings from 48 sires and 77 dams with a nested dam-sire structure. Shrimps were individually weighed at an average age of 130 days post-hatching. BW phenotypic mean (S.D.) was 18.2 (2.4) g, with values ranging from 8.4 to 30.0 g. Data were analyzed using univariate and multivariate models that considered BW within location by density pond environment as a different trait and included or not a common full-sib effect (c). The multivariate animal model included fixed effects of days from hatching and sex. For univariate models that included c effects, BW heritability (S.E.) estimates ranged from 0.24 (0.14) to 0.35 (0.18) across environments (heritability was zero in one environment). For multivariate models (excluding the environment with zero heritability) the heritabilities increased and ranged from 0.37 (0.06) to 0.45 (0.09). Standard errors of heritabilities and c effects were both drastically reduced in the multivariate analysis. Pairwise genetic correlations between environments were from 0.80 (0.08) to 0.86 (0.04). These differences may be indicative of genotype-environment interaction for BW at 130 days post-hatching. Statistical problems found to separate c from additive genetic effects both in univariate models were reduced using multivariate models. Correlation between family raw phenotypic means and family BV means from the multivariate analysis was 0.93 indicating a rather low risk of miss selecting superior families if BLUP solutions were neglected using replicated environment data. It is also concluded that use of incorrect statistical models or unreplicated data may lead to biased or inaccurate estimates of genetic parameters in shrimp breeding programs.     

Controlling the growth of a cyanobacterial contaminant, Synechoccus sp., in a culture of Tetraselmis chui (PLY429) by varying pH: Implications for outdoor aquaculture production

In aquaculture, large volumes of phytoplankton are often grown outdoors to reduce costs. However, growing microalgae in an environment not as well-controlled as in the laboratory can lead to unwanted phytoplankton, including cyanobacteria, contaminating a culture. A cyanobacterial contaminant was isolated from an outdoor culture of Tetraselmis chui (PLY429) at the Milford Laboratory. This study investigated the growth of PLY429 and the cyanobacterium in pure cultures and a mixed culture in a pH range of 6.5-9.5. The division of PLY429 was greater at a pH range of 7.0-8.0; whereas, for the cyanobacterium, higher growth was obtained at pH 8.0-9.0. Results from combined cultures of PLY429 and the cyanobacterium grown at various pHs indicated that maintaining pH near 7.1 yields higher growth of PLY429 than those of the cyanobacterium. These findings suggest that controlling pH may reduce the population of a cyanobacterial contaminant in an aquaculture feed culture.     

Investigating fragment size for culturing reef-building corals (Porites lobata and P. compressa) in ex situ nurseries

Culturing small propagules of coral has the potential for high yield with low environmental impact, provided that mortality is low. This study investigated the size-specific mortality and growth of minute fragments (as small as 5 polyps) of two of the most abundant reef-building corals in Hawaii (Porites lobata and P. compressa). Two ex situ nursery systems differing in cost, design complexity, and labor intensity were compared. The first nursery experiment lasted for four months in a large tank with high water motion/surge and extensive manual cleaning to remove competitive algae. The corals were then transferred to a simple low-flow tank containing sea urchins (Tripneustes gratilla) and reduced cleaning, where they were grown for six more months (Nursery II). ‘Nursery I’ resulted in 92% of P. lobata and 73% of P. compressa fragments surviving and nearly doubling in area, in spite of a brief infestation by a nudibranch (Phestilla sibogae) that primarily fed on larger P. compressa fragments. There was a significant positive relationship between fragment size and growth rate, and survivorship was significantly different between donor colonies (genets), but there was no evidence of size-specific mortality. ‘Nursery II’ on the other hand had clear size-specific mortality and higher urchin damage for smaller fragments, resulting in moderate survivorship (78% for P. lobata and 76% for P. compressa), and only a slight increase in the total area covered by coral tissue. Fragments larger than 3 cm² were undamaged and had the highest survival and growth rates. This study illustrates how size-specific mortality can be reduced by ex situ nursery conditions.     

Recruitment and effects of Discocotyle sagittata (Monogenea) infection on farmed trout

Farmed rainbow trout Oncorhynchus mykiss and brown trout Salmo trutta were monitored over 3 years for infection with the blood-feeding gill fluke Discocotyle sagittata. Parasite transmission is seasonal: new infections take place during summer/autumn, and transmission is generally negligible during winter/spring. There are 2 sources of infection for naïve fish-of-the-year: limited invasion when fish are in the raceways by riverborne larvae originating external to the farm; and internally, within the farm, when 0+ fish are transferred to ponds previously occupied by older cohorts of infected fish. Thereafter, infection levels continue to increase in rainbow trout primarily through transmission within the farm. Prevalence rose to 100% in 1+ fish by the end of their second summer. In O. mykiss, mean abundance reached 194 worms/host for 1+ fish (up to 489 worms/host) and 160 worms/host for 2+ fish. By contrast, in S. trutta, parasite prevalence never exceeded 85% and, after the first year's invasions, infection levels decreased over time: in 1+ and 2+ brown trout, parasite mean abundance was < 4 (maximum 15) worms/host. We present evidence of the detrimental effects of D. sagittata on the host: high burdens are associated with pale gills, decreased body condition and host mortality. Parasite burdens become overdispersed during the warmer part of the year, as prevalence and mean abundance increase. However, the degree of parasite overdispersion decreases over winter; we cannot distinguish whether decreased aggregation is due to parasite losses from infected fish (including immune-mediated parasite mortality) or parasite-induced host mortality.     

Chicken-derived IgY recognizes developing and mature stages of Loma salmonae (Microsporidia) in Pacific salmon, Oncorhynchus spp.

A polyclonal IgY preparation, purified from yolk of immunized hens, is shown to be useful for the detection of Loma salmonae in Pacific salmon. Purity of the IgY was confirmed by SDS-PAGE and western blots of yolk protein extracts. Reactivity of the IgY preparation to L. salmonae was determined through indirect-fluorescent antibody tests on purified spores and by immunohistochemistry. Developing xenomas in histological sections of heart and gill tissue of infected chinook salmon (Oncorhynchus tshawytscha) were visualized as early as 4 weeks post-exposure (PE). Non-lethal detection of L. salmonae was obtained by indirect-fluorescent microscopy of acetone-fixed gill smears. The IgY preparation was shown to react with another Loma sp., L. branchialis. In contrast, no reactivity was observed with other microsporidian parasites of fish such as Pseudoloma neurophilia and Glugea anomala. Yolk derived IgY may be useful in the detection of other fish pathogens.     

Oxytetracycline (OTC) uptake following bath treatment in gilthead sea bream (Sparus aurata)

The uptake and elimination profile of oxytetracycline (OTC) following a prolong bath treatment in gilthhead sea bream (Sparus aurata) were investigated in this study. The bath experiment was carried out using a OTC concentration of 50 μg/ml for 24 h at 17-18 °C water temperature. Plasma and muscle fish samples were analysed at 1, 3, 6 and 24 h during and at 1, 2, 3, 4 and 6 d following the bath. Detectable OTC levels were revealed only at the end of bath treatment (24 h) in examined tissues of gilthead sea bream, where drug concentration was measured to be as low as 0.096 and 0.047 μg/g or ml in muscle plus skin and plasma, respectively. The findings of the present study indicate that OTC bath treatment under this dosage schedule is unlikely to confront systemic bacterial infections.     

Early development of the shortfin silverside Chirostoma humboldtianum (Valenciennes, 1835) (Atheriniformes: Atherinopsidae)

The shortfin silverside Chirostoma humboldtianum has been considered for culture in Mexico, but success has been limited by a poor knowledge of its early development. First synthesis of the early development of the shortfin silverside is presented to determine conditions suitable for rearing. Brooder maturation was induced through photothermal cycles. C. humboldtianum ova were fertilized in vitro. The eggs were incubated in reconstituted water (160-180 mg/L CaCO₃) at 18 °C and 5 gm of NaCl per litre. During the hatching day, 300 shortfin silversides were stocked and followed up until metamorphosis in order to establish the timing of exogenous feeding, changes in food type, growth and development during critical periods for survival, according to the theory of saltatory ontogeny. Free embryos hatched 12 days after fertilization at 18 °C. First critical point for survival is the beginning of exogenous feeding. Free embryos started mixed feeding on day four of post-hatching (dph), point of no-return was presented towards the end of mixed feeding on 6 dph, larval period began at six (dph) when the anus is opened, and metamorphosis to juvenile was presented at 65 dph with a SL of 19.34 ± 2.28 mm, when scales and fins were well developed. Differences in growth between periods were detected: free embryos growth slower than larvae but mouth size depicted a larger growth rate in the former. Cephalic length and mouth size were negatively related to standard length in embryos and larvae. Mouth size was positively related to cephalic length in free embryos but negative in larvae. Results suggest that during the free embryo phase, growth priorities are directed to the development of apparatuses and systems; whereas, during the larval period, energy is directed to growth in length, mouth size and development of fins, which allows them to increase their swimming velocity, grants them a greater capacity to obtain exogenous food and, in consequence, increases fitness for survival.     

Distribution of a fish pathogen Listonella anguillarum in the Japanese flounder Paralichthys olivaceus hatchery

The present study was undertaken to investigate the distribution of Listonella anguillarum in a Japanese flounder (Paralichthys olivaceus) hatchery. A total of 2704 isolates were obtained from the developing fish, live diets and artificial feeds of Japanese flounder and their rearing water, 439 of which were identified as L. anguillarum by the combining incubation on thiosulfate-citrate-bile salt-sucrose (TCBS) agar at 35 °C overnight with polymerase chain reaction (PCR) detection for the VAH1 hemolysin gene. L. anguillarum was detected in all seven rotifer samples, with densities of 2.5 × 10³ to 4.6 × 10⁶ colony forming units (CFU) g^− 1. Both the analyzed samples of Nannochloropsis oculata contained this bacterium at densities of 1.6 × 10⁴ to 1.4 × 10⁵ CFU g^− 1. L. anguillarum was detected in only one of four samples of Artemia nauplii with a density of 4.8 × 10⁵ CFU g^− 1 (35%) and it was not detected in the two analyzed artificial feed samples. L. anguillarum was detected in 11 of 18 specimens of larval and juvenile Japanese flounder at densities of 5.0 × 10¹ to 7.4 × 10⁵ CFU g^− 1, while it was not detected in the two analyzed egg specimens of Japanese flounder. These results indicate that L. anguillarum associated with the developing Japanese flounder is likely derived from rearing water and live diets such as rotifers. Further, it is strongly suggested that L. anguillarum is a transient bacterium of the intestinal microflora for the Japanese flounder but is a permanently indigenous one for the Japanese flounder hatcheries.     

Feed preferences of Salminus brasiliensis (Pisces, Characidae) larvae in fish ponds

Feed preferences of dorado, Salminus brasiliensis, in the early stages of life was studied in larvae, stocked 5 days after hatching in three earthen commercial ponds (810 m², 900 m² and 1480 m²) at a density of 30 m^− 2. Larvae were fed on natural food from ponds supplemented with a commercial ration (40% crude protein), and for 12 days fish samples were taken daily in the morning and in the afternoon from each pond to analyze the food items present in the diet. Zooplankton available in the environment was also analyzed. Feeding was intensive since 94% of the larvae guts examined contained food. Since the commercial diet was not found in the dorado guts, the natural diet dominated the artificial food, but the cannibalistic behavior typically found in laboratory conditions was not observed. Insect larvae and cladocerans were the main prey items found in the gut of the larvae. In fish ponds dorado larvae can be considered a generalist feeder, since small amounts of different food items were consumed, but they tend to specialize in some items, according to their developmental stage.     

Suitability of the copepod, Acartia clausi as a live feed for Seabass larvae (Lates calcarifer Bloch): Compared to traditional live-food organisms with special emphasis on the nutritional value

Though artificial propagation of Asian seabass Lates calcarifer (Bloch) in captivity through induced breeding techniques is standardized under Indian conditions, larval and nursery rearing techniques including suitable nursery feeds have to be standardized to obtain better survival and growth. Feeding experiments in triplicate were conducted to evaluate the suitability of the marine copepod Acartia clausi as live prey for fourteen day-old seabass larvae (6.53 ± 0.06 mm; 8.58 ± 0.33 mg) and compared with the traditional live prey, rotifers and Artemia nauplii. While A. clausi and rotifers were mass produced using algae Isochrysis galbana, Chaetoceros affinis and Chlorella marina, Artemia nauplii were produced using cysts. Nutritional quality of cultured copepods was evaluated based on the proximate composition, amino acid and fatty acid composition, and compared with that of rotifers and Artemia nauplii. Proximate composition varied significantly (P < 0.05) among the different live feeds. A. clausi showed higher protein (63.12%) and lipid (16.65%) content than Artemia nauplii and rotifers. Total essential amino acids content was 2% lower in A. clausi compared to that in Artemia nauplii. Fatty acid profiles of the live feed organisms showed that A. clausi is a rich source of n − 3 fatty acids. The total n − 3 fatty acid content of A. clausi was 33.94%. Length, weight overall weight gain and survivorship were significantly (P < 0.05) different among the dietary treatments, and weight gain was comparatively higher in A. clausi fed larvae. Survival of seabass larvae fed A. clausi was obtained highest as 58.13% against the lower values of 39.93% and 41.62% in larvae fed rotifer and Artemia nauplii respectively. Final carcass composition of the larvae of L. calcarifer fed different live-food organisms showed significant differences (P < 0.05) among the dietary treatments. The fatty acid composition of the dietary treatments was reflected to a certain extent in the fatty acid composition of the seabass larvae. The present investigation revealed the nutritional value of calanoid copepod and thus underlining its usefulness as a suitable live-food organism for rearing larvae of the commercially valuable Asian seabass.     

Accuracy of pairwise methods in the reconstruction of family relationships, using molecular information from turbot (Scophthalmus maximus)

Many estimators and algorithms have been developed to infer the genealogical relationships from molecular marker data when there is no pedigree information. Most pairwise methods provide estimates in a continuous range that needs to be converted into genealogical relationships (namely full-sibs, half-sibs and unrelated) if there is a previous knowledge of the population structure. Transformations are usually based on arbitrary thresholds, but the possibility of correcting the coancestry estimates via explicit pedigree reconstruction methods has been suggested. Using molecular data for ten highly polymorphic microsatellite loci on a population of turbot (Scophthalmus maximus) with a known genealogy, the behaviour of four pairwise marker-based coancestry estimators and the molecular coancestry has been evaluated. The population consisted on 138 families with 4 full-sib individuals each and one family with 8 full-sib individuals (up to 15 half-sib families were also present due to the sharing of parents between some of the full-sibs families). Our results suggested that transforming the pairwise estimators and the molecular coancestry to family relationships through the establishment of thresholds performs slightly better than the explicit pedigree reconstruction method, when accuracy is measured in a pairwise basis. However, if joint relationships between more than two individuals were tested at a time, the threshold methods led to a high percentage of incongruous triads of full-sib individuals, with Mendelian incompatibilities appearing in congruous full-sib families (more than 70% and 60% in our study, respectively). The explicit pedigree reconstruction approach, due to its nature, is free from such problems. Therefore, the pedigree reconstruction approach seems to be a valuable tool to provide a congruent and compatible pedigree when the pairwise marker-based coancestry matrices or the molecular coancestry need to be transformed.     

Morphological characterization of larval stages and first juvenile of the freshwater prawn Cryphiops caementarius (Molina, 1782) (Decapoda: Palaemonidae) under laboratory conditions

Research has been in progress for several years on various aspects of the biology and ecology of the freshwater prawn Cryphiops caementarius, an inhabitant of rivers in northern Chile. The commercial value of this prawn fomented the accomplishment of studies on its reproduction and development with the aim of producing juveniles under controlled conditions, to be followed by growout to commercial size in managed culture systems. The present study describes larval culture of this species from eggs of gravid females obtained in the field, from the first developmental stage (Zoea I) through the first juvenile stage. The larvae were cultured at 25 °C in UV sterilized water at variable salinities based on the requirements of the developmental stages. Larvae were fed with Nannochloris, Isochrysis and Artemia nauplii as required. This report describes in detail the 18 larval stages of this prawn, as well as its first juvenile form.     

Comparison of diagnosis techniques for the protozoan parasite Bonamia ostreae in flat oyster Ostrea edulis

In recent years, the cultured production of the flat oyster Ostrea edulis has suffered a dramatic decrease in Europe partially attributed to the protozoan parasite Bonamia ostreae, the causative agent of the Bonamiosis. In this paper the results of a PCR assay for the diagnosis of B. ostreae were compared with those obtained using two classical methods of diagnosis recommended by the OIE (Office International des Epizooties) and the European Union, histology and cytology. The same samples were analyzed by two different laboratories, showing that the results obtained with the PCR method have high sensitivity and good correlation between laboratories. This method is cheaper and faster than histopathology and cytology, with no need of specifically trained personnel to perform the diagnoses. It is appropriate for fast screening of stocks of great numbers of oysters.     

Identification and phylogenetic analysis of Vibrio vulnificus isolated from diseased Trachinotus ovatus in cage mariculture

In October 2005, an infectious disease occurred as outbreaks of high mortality within one week, which was responsible for important economic losses in intensive culture of Trachinotus ovatus around the Gulf Coast of Yangjiang city (Guangdong Province, China). The present study documented the causative agent of the diseased fish suffering from external haemorrhages and ulcers, haemorrhagic gills, livers and intestine. The strain was isolated from the diseased fish by ZoBell 2216 E agar and confirmed the pathogenicity by challenge experiments. Biochemically, the strain showed properties and biochemical characteristics similar to Vibrio vulnificus, with the exception of several atypical biochemical characteristics, especially the sucrose-positive. Meanwhile, sequencing of the 16S rRNA gene and the hemolysin gene revealed that the isolated strain was highly homogeneous with V. vulnificus. To sum up, the isolated strain was confirmed as V. vulnificus on the basis of phenotypic characteristics, phylogenetic analysis of the 16S rRNA sequences and sequence analysis of the hemolysin gene. To our knowledge, this is the first report on the V. vulnificus infection in T. ovatus.     

In situ studies on the bioaccumulation of microcystins in the phytoplanktivorous silver carp (Hypophthalmichthys molitrix) stocked in Lake Taihu with dense toxic Microcystis blooms

The phytoplanktivorous silver carp is an important biomanipulation fish to control cyanobacterial blooms and is also a food fish with the greatest production in China. The accumulation of the hepatotoxic microcystins (MCs) determined by LC-MS in various organs of silver carp was studied monthly in Lake Taihu dominated by toxic Microcystis aeruginosa. Average recoveries of spiked fish samples were 78% for MC-RR and 81% for MC-LR. The highest content of MCs was found in the intestine (97.48 μg g^− 1 DW), followed by liver (6.84 μg g^− 1 DW), kidney (4.88 μg g^− 1 DW) and blood (1.54 μg g^− 1 DW), and the annual mean MC content was in the order of intestine > liver > kidney > blood > muscle > spleen > gallbladder > gill. Silver carp could effectively ingest toxic Microcystis cells (up to 84.4% of total phytoplankton in gut contents), but showed fast growth (from 141 g to 1759 g in 1 year in mean weight). Silver carp accumulated less microcystins in liver than other animals in the same site or other fish from different water bodies at similar level of toxin ingestion. There was possible inhibition of the transportation of the most toxic MC-LR across the gutwall. Muscle of silver carp in Lake Taihu should not be consumed during period of dense Microcystis blooms while viscera were risky for consumption in more months.     

Chilled storage of white shrimp (Litopenaeus vannamei) spermatophores

Chilled storage of spermatophores from white shrimp (Litopenaeus vannamei) is needed to generate a consistent and reliable supply of spermatozoa for domestication purposes. The objective of this study was to develop a protocol for the chilled storage of white shrimp spermatophores and to evaluate bacterial propagation during such storage. In the first experiment, spermatophores were immersed in four extenders, mineral oil, Ringer's solution, phosphate buffer and 0.85% NaCl, and stored at low temperature (2-4 °C) for 35 days. Characteristics of preserved spermatophores changed the least and viable sperm was highest when spermatophores were stored in mineral oil. Spermatophores preserved with mineral oil appeared morphologically normal. Bacillus circulans, Staphylococcus hominis and S. lugdunensis, S. sciuri, S. xylosus and Micrococcus spp. were identified as the predominant bacteria during chilled storage, and total bacterial counts gradually increased during the experiment. A second experiment investigated the effect of antibiotic on chilled storage. Spermatophores were preserved in only mineral oil or mineral oil with 0.1% penicillin-streptomycin. These were evaluated for changes in external morphology of spermatophores, sperm viability and total bacteria count every week during a 35-day experimental period. Percentages of viable sperm (69.5 ± 3.9%) were significantly higher (P < 0.05) among spermatophores preserved in mineral oil with 0.1% antibiotic compared with those preserved only in mineral oil (57.7 ± 3.4%) over 35 days. The number of total bacteria in the treatment with mineral oil ranged between 28.3 ± 4.8 and 2416.7 ± 299.4 CFU/g, but in mineral oil containing antibiotic bacteria were undetectable. This study suggests that chilled storage of spermatophores is a feasible approach for the management and spawning of white shrimp broodstock.