Physio-Chemical Characteristics of Seminal Plasma and Development of Media and Methods for the Cryopreservation of European eel Sperm

The high sperm density, together with the short spermatozoa swimming time, makes European eel sperm manipulation and assessment for quality difficult. Two diluting media (K15 and K30) previously designed for Japanese eel sperm were tested. After 24 h, European eel sperm showed significant reduction in the percentage of motile spermatozoa after activation and different motility parameters (VAP, angular velocity; VCL, curvilinear velocity; VSL, straight line velocity; BCF, beating cross frequency), concluding that these media are not suitable to preserve the sperm of this species. After a hormonal treatment to induce spermiation, sperm volume, density and motility were recorded at weekly samplings. The variation of the osmolality (325–330 mOsm kg⁻¹), pH (8.4–8.6) and the ionic composition (concentration of Na⁺, K⁺, Mg²⁺ and Ca²⁺) of the seminal plasma were registered. Physio-chemical results were related with sperm quality throughout the treatment, to determine which must be the suitable characteristics of one extender for the sperm of this species, and to find the best conditions to obtain suitable cryopreservation media for European eel sperm. K⁺ concentration increased, while Ca²⁺ and Mg²⁺ concentrations showed a progressive reduction in correlation with the sperm quality improvement. Na⁺ showed a decreasing, but not significant tendency. P1 and P2 freezing media were designed considering the physio-chemical parameters as well as the ionic composition shown by the best quality sperm samples, and then compared with the previously described solutions, TNK and K30. Sperm quality was determined, checking the percentage of motile spermatozoa and motility parameters using computer-assisted sperm analysis (CASA) software. Samples were frozen after dilution (1:5, 1:20, 1:100) in different freezing media supplemented with 10% dimethyl sulfoxide (DMSO). After thawing, samples frozen with low dilution ratio (1:5) in TNK and P1 media showed higher, although not significant, spermatozoa survival (35.5 ± 14.5 and 36.6 ± 6.7%). The addition of l-α-phosphatidylcholine to the media seems to have a positive effect, as reported in the Japanese eel.     

Effect of Senna occidentalis seeds on immunity in broiler chickens

This study investigated possible immunotoxic effects of Senna occidentalis (So) seeds incorporated in broiler chicken rations at different concentrations (0.0%, 0.25%, 0.50% and 0.75%), for 28 or 42 days. We evaluated innate immune function (macrophage activities of spreading, phagocytosis, peroxide and nitric oxide production) and acquired immune function (humoral and cellular immune responses), as well as lymphoid organ weights and pathology. There was enhanced macrophage activity, as hydrogen peroxide production increased (P < 0.05) in cells of birds given 0.75%So, but there were no other pro-inflammatory effects. Birds receiving 0.75% of So in ration for 42 days gained less weight (P < 0.01), and showed a decrease in relative weight of the bursa of Fabricius (P < 0.05) and spleen (P < 0.01). In addition, morphological changes were also noted in these lymphoid organs, with depletion of lymphoid cells on the spleen and bursa of Fabricius, resulting in lower relative weight of both lymphoid organs. No impairment of humoral immune response against Newcastle disease and in cellular immune response after a phytohaemagglutinin challenge was found. It is probable that mitochondrial damage and related apoptosis may be responsible for the enhanced peroxide production and the reduced relative weight of the bursa of Fabricius and spleen.     

On the independence between clupeid stocks and their recruitment

The statistical independence between the reproductive stock and its corresponding recruitment to a fishery was tested for 43 stocks of clupeids in the Ransom Myers database by using the chi-squared test of a contingency table. The method was validated by simulating stock and recruitment (SR) contingency tables for data close to hypothetical SR curves that represent several known phenomena. The clupeid SR data were grouped by the criteria of percentile points to build the contingency tables. The independence test of the clupeid stocks was rejected in only 15 cases, thus the independence hypothesis was sustained in the remaining 28 cases. This demonstrates that fitting an SR model in these clupeid stocks is not justified because their abundance could be affected by several causes other than reproductive stock fluctuations.

     

Effect of dietary <Emphasis Type="SmallCaps">l</Emphasis>-tryptophan on cannibalism,survival and growth in pikeperch <Emphasis Type="Italic">Sander lucioperca</Emphasis> (L.) post-larvae

The effect of supplemented commercial diets with crystalline l-tryptophan (TRP—5, 10 and 20 g TRP kg^?1) on cannibalism, survival and growth parameters of pikeperch post-larvae (Sander lucioperca) was evaluated. Fifteen-day-old pikeperch larvae (mean weight—6.8 mg) were reared during the next 28 days (20.5 °C, 16L:8D) in glass aquaria in a recirculating aquaculture system. The enzyme-linked immunosorbent assay showed that TRP-supplemented diets were effective in increasing the levels of serotonin (5-HT) in the body tissue of pikeperch. TRP supplementation resulted in a slight decrease in both types of cannibalism, although the reduction in cannibalism impact did not amount to more than a few percent. TRP treatment had no significant influence on the final survival of pikeperch post-larvae (ranged from 20.1 ± 12.4 to 29.0 ± 12.9 %). However, contrary to the earlier studies conducted on other fish species, no significant difference in the growth rates and feeding behavior of pikeperch between TRP-fed and control group were found. The final body weight and growth rate ranged from 0.211 ± 0.014 to 0.243 ± 0.016 and from 12.19 ± 0.38 to 12.76 ± 0.35 % day^?1, respectively. To our knowledge, this is the first study reporting the effects of TRP supplementation on the cannibalism–survival–growth relations in fish in the post-larval stage.     

Growth and survival of the juvenile yellowleg shrimp Farfantepenaeus californiensis cohabiting with the green feather alga Caulerpa sertularioides at different temperatures

This study was performed to determine the growth and survival patterns of the juvenile yellowleg shrimp Farfantepenaeus californiensis cohabiting with the green feather alga Caulerpa sertularioides over a range of temperatures (18, 22, 25, 28 and 32°C) found throughout the year in the Gulf of California. From an initial weight of 46 ± 10 mg, shrimp cohabiting with green feather algae increased in weight by an average of 0.57 (±0.07), 5.44 (±0.45), 6.36 (±0.37), 7.66 (±0.26) and 5.96 (±0.59) g at 18, 22, 25, 28 and 32°C, respectively, whereas shrimp grown without seaweed increased in weight by an average of 0.28 (±0.026), 2.66 (±0.24), 3.15 (±0.49), 4.00 (±0.67), and 3.54 (±0.09) g respectively. Statistically significant differences were observed for the growth rates of juvenile yellowleg shrimp in the presence of C. sertularioides and peak growth was observed at 28°C in experiments with macroalgae. No significant differences were observed for shrimp survival except at 32°C without macroalgae, where survival was lower. These results suggest a beneficial and growth‐promoting effect of cohabitation with live C. sertularioides on F. californiensis.     

Ovulation induction in jundiá (Rhamdia quelen,Heptapteridae) using carp pituitary extract or salmon GnRH analogue combined with dopamine receptor antagonists

Jundiá (Rhamdia quelen) is an important species for aquaculture in temperate and subtropical climates. In the present study, the results of different forms of GnRH analogue treatments on ovulation in jundiá have been compared with those from treatment with carp pituitary. Seven out of eight females ovulated in groups treated with carp pituitary (4.0 mg kg^?1 BW) and Ovaprim (0.5 mL kg^?1 BW). Injection with sGnRHa (10 μg kg^?1 BW) combined with metoclopramide (20 mg kg^?1 BW) caused a significantly lower rate of ovulation with three out of eight females responding. None of the fish injected with sGnRHa (10 μg kg^?1 BW) alone ovulated. Mean PGSI [(weight of stripped egg mass/BW of the female before stripping) × 100] and mean fertilization rate values were high and similar between treatment groups. The obtained results indicate that in jundiá, there is a strong dopamine inhibitory tone on gonadotropin secretion. The gonadotropin releasing activity of sGnRHa can be potentiated by dopamine receptor antagonists and the combined treatment is effective in inducing ovulation. In this regard, domperidone proved to be more potent than metoclopramide.     

Tissue composition,blood biochemistry and histology of digestive organs in Senegalese sole (Solea senegalensis) juveniles fed diets containing different plant protein ingredients

The aim of this study was to determine the impact of diets with different plant protein ingredients on proximate composition, tissue metabolites and tissue fatty acid composition, as well as hepatic and intestinal histological modifications in Senegalese sole (Solea senegalensis). Fish (21.5 ± 2.8 g body weight) were fed six isonitrogenous and isoenergetic diets during 11 weeks. A control diet containing fish meal as the main protein source was compared with five experimental diets replacing 30% fish meal protein by different plant protein sources: soybean meal (SBM), soybean protein concentrate (SPC), soybean protein isolate (SPI), wheat gluten meal (WGM) or pea protein concentrate (PPC). The inclusion of different plant protein did not significantly affect growth and proximate composition of fish. The hepatosomatic index was not significantly different to the control group; however, utilization of WGM significantly increased hepatocyte size. Plasma protein values and muscle triglycerides were influenced by the inclusion of SBM and WGM in the diets respectively. Feeding fish on SBM, WGM and PPC diets significantly enhanced n‐6 fatty acid levels in muscle, particularly linoleic acid. None of the plant protein ingredient used in the diets decreased arachidonic, eicosapentaenoic as well as docosahexaenoic acid values in liver and muscle. Overall, histological studies did not reveal the existence of any intestinal alterations induced by the inclusion of different plant proteins. Despite moderate changes produced by SBM, SPC and WGM, inclusion of dietary plant protein ingredients has no major impact on growth, tissue and blood biochemistry, fatty acid profile and gut integrity of Senegalese sole juveniles.     

Non‐invasive fast real‐time PCR assay for detection of the enteric parasite Enteromyxum scophthalmi in cultured turbot (Scophthalmus maximus L.)

Enteromyxum scophthalmi is a myxozoan parasite that causes severe parasitic diseases in cultured turbot affecting mainly the intestine of the host. It is characterized by producing acute enteritis, starvation and eventually death. Current diagnosis of E. scopthalmi use traditional techniques, based on the identification of the morphology of the parasite. These techniques take extended time to be carried out and do not favour the adoption of control measure at turbot farms and require the sacrifice of fish. This study develops a fast real‐time PCR molecular tool for the detection of E. scophthalmi in infected farmed turbot. This methodology is applicable for routine controls on the farm at every stage of the parasite infection. Results of the study demonstrate the robustness, specificity, efficiency and reliability of the technique. In addition, this study also provides a non‐invasive procedure of sampling through swaps. This allows control, prevention and diagnosis of the parasite infection at turbot farms while maintaining the welfare of the cultivated fish and avoiding sacrifice of the fish sampled.