Multiple paternity in reproduction of European eel Anguilla anguilla (L. 1758) by artificial mixing of different sperm in equal volumes

Because of its complex life cycle and due to multiple stress factors, the European eel is suffering a dramatic decline and has been declared Critically Endangered by the IUCN. A possible way to address this problem could be relieve the pressure on natural stocks by implementing its mass production by artificial breeding protocols. A previous study based on direct observation and parentage assignment underlined the presence of an allegedly hierarchic structure among European eel males in semi-natural mating conditions, with a consequent bias in F1 proportion assigned to each one. The aim of the present study is to attest if a different fertilization protocol based on the artificial mixing of female eggs with different males’ milt in equal volume (1 F:4 M) could represent a solution to the disproportion observed in semi-natural conditions. For this purpose, six families of European eels were generated, and 10 species-specific microsatellite loci were used to infer offspring composition by paternity assignment on 280 samples. Due to the asynchronous ovarian development of female European eels, the per- centage of fertilized eggs for each female showed a great variability, ranging from 4.70% to 94.50%. A pro- portion of 94.02% of genotyped offspring were assigned with high confidence to their true parents. As regard males’ fertilization pattern, no substantial differences from natural mating were observed: a single male accounted for most offspring, which was just mostly composed of full sibs. Concluding, the obtained results suggest that the admixture of an equal volume of different males’ milt seems to contribute to the single-locus genetic variability (observed heterozygosity higher than expected in 7 out 9 loci), but it is not sufficient to ensure all the males the same chance to transmit their gene pool, and new fertilization strategies must be developed

Evaluation of reproductive performances of the common octopus (Octopus vulgaris) reared in water recirculation systems and fed different diets

The reproductive performance of Octopus vulgaris broodstocks fed two different diets (mixed fish [F group, BW 1,048.14 g] or mixed crustaceans [C group, BW 998.44 g]) was analyzed using an experimental recirculating aquaculture system consisting of a tank equipped with spawning and incubation chambers. A total of 8 females (F1–4; C1–4), and 8 males (M1–M8) were selected. DI of the C group females was significantly (p < 0.05) higher (3.0 ± 0.29%) than the F group (2.16 ± 0.67%). SGR in C group was significantly higher (1.43 ± 0.12%) than the F group (1.18 ± 0.25%). Egg clusters, number of clusters, number of clusters/kg BW, and total length were more favorable in the C group than the F group. The number of clusters/kg BW of C females was 2.5 times higher than that of F females (78.1 ± 6.5 vs 31.1 ± 13.3). The total eggs number, number of eggs/cm, number of eggs/kg BW in the C group were significantly (p < 0.05) higher compared with the F group; the number of eggs/kg BW and paralarvae/kg BW were 5 times higher in the C group (115,928 ± 12,513 C vs 22,109 ± 7912 F and 114,953 ± 12,591 vs 20,729 ± 7104, respectively). Hatching rate of the C group was significantly (p < 0.05) higher compared to the F grou

Comparison of zootechnical performances, welfare condition and quality traits of hybrid striped bass (Morone chrysops x Morone saxatilis) reared in different Italian farms.

Growth performances of sunshine bass (initial m.b.w.=200\ub150 g) were examined in 3 different rearing conditions: concrete square basins (HSB-1); raceways (HSB-2); natural ponds (HSB-3). Fish received the same extruded feed (prot. 48.8%; lip. 17.8%). The trial lasted 16 months in HSB-1 and HSB-2 and 12 months in HSB-3 due to unfavourable welfare status and low growth performances. At harvest (16 months), sunshine bass reached a satisfactory mean weight without significant differences between groups. HSB-2 fish were longer with a smaller circumference, probably due to different hydrodynamics of the raceway. Glucose, cholesterol, triglycerides, protein, lactic dehydrogenase (LDH), aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), creatine kinase (CK), glycogen of plasma or tissue were not affected by rearing conditions as well as the proximate composition and the fatty acid profile of the fillet

A methodological approach to assess nutrient requirements of heavy pigs in Italy

This paper reports the results of the work of the "Nutrient requirements of heavy pigs" ASPA Commission. The Commission work was mainly focused on the problem of identifying a good and flexible methodology for establishing nutrient requirements of heavy pigs in Italy, in a situation where the major part of the pigs are used for ham and other typical products. Nutrient allowance was considered to be a variable open to manipulation, since its optimal definition depends on the specific circumstances of each single production unit. It appeared that the most logical way to account for the many interactions involved was to integrate available information using computer simulation techniques. A methodology based on the following steps was proposed and analyzed: i) selection of a mathematical model to describe the chemical growth of pigs over time as result of the interactions of genotype, feed and climate; ii) developing suitable equations to predict the quality parameters of the carcass and, if possible, of the single cuts from the body's chemical status and implementation of these equations in the model; iii) identification of production targets and use of the model as a tool to simulate the effect of different genotypes, feed and climate on the productive and the economic results. A model strongly rooted in a theoretical framework, simple enough to be applied in practice and requiring a limited number inputs was selected from literature. From a preliminary evaluation, performed by using experimental calorimetric data, it was concluded that the model was potentially able to accurately predict the chemical growth of pigs under different conditions of feeding and climate. The major lack of knowledge regarded the values of genotypic descriptors of the national pig populations, the evaluation for which some simple experimental protocols are described in literature. Other lack of knowledge involved relationships to predict the anatomical growth and the evolution of quality parameters of carcass and cuts from the chemical composition of the body. If a correlation between the chemical composition of the pig's body and production targets is confirmed, then a link between the farmer and the industry can be established and the model could be used to identify more convenient feeding strategies. The conceptual basis and the way to assess nutrient requirements of heavy pigs have been clearly identified

Comparative evaluation of the effects of different activating media and temperatures on European eel (Anguilla anguilla) sperm motility assessed by computer assisted sperm analysis

The European eel is a critically endangered teleost fish with very poor success rate for captive breeding and artificial reproduction. Therefore, to support its conservation, new strategies are needed to ensure fertilization. Objective analysis of sperm motility may be critical as it potentially represents one of the most important reproductive quality parameters. Spermatozoa acquire motility once in contact with hyperosmotic solutions as saltwater, yet the exact mechanisms and the role of temperature are still to be clarified. The main aim of the study was to assess the effects of 3 activating media (artificial sea water, tank water and commercial Actifish®) at 4 and 20 °C on sperm motility, by means of computer assisted sperm analysis. Secondary aim was to test 2 different concentrations of Actifish® mimicking sea water pH/osmolality, at 4 °C. The results suggested how both temperature and activating media have effects on spermatozoa motility and kinematics, with temperature mainly acting upon interaction with the media type. The samples activated with tank water at 20 °C showed the poorest motility outcomes (mean 38.1%), while the ones activated with Actifish® diluted 1:4 and artificial sea water, at 4 °C, the highest (means 51.8 and 51.5% respectively). Additionally, diluting Actifish® to reach same pH and osmolality of seawater led to worse motility outcomes, suggesting that composition may be the critical factor for activation rather than osmolality itself

A methodological approach to assess nutrient requirements of heavy pigs in Italy

This paper reports the results of the work of the "Nutrient requirements of heavy pigs" ASPA Commission. The Commissionwork was mainly focused on the problem of identifying a good and flexible methodology for establishing nutrient requirements of heavy pigs in Italy, in a situation where the major part of the pigs are used for ham and other typical products. Nutrient allowance was considered to be a variable open to manipulation, since its optimal definition depends on the specific circumstances of each single production unit. It appeared that the most logical way to account for the many interactions involved was to integrate available information using computer simulation techniques. A methodology based on the following steps was proposed and analyzed: i) selection of a mathematical model to describe the chemical growth of pigsover time as result of the interactions of genotype, feed and climate; ii) developing suitable equations to predict the qualityparameters of the carcass and, if possible, of the single cuts from the body's chemical status and implementation ofthese equations in the model; iii) identification of production targets and use of the model as a tool to simulate the effectof different genotypes, feed and climate on the productive and the economic results. A model strongly rooted in a theoreticalframework, simple enough to be applied in practice and requiring a limited number inputs was selected from literature.From a preliminary evaluation, performed by using experimental calorimetric data, it was concluded that the modelwas potentially able to accurately predict the chemical growth of pigs under different conditions of feeding and climate.The major lack of knowledge regarded the values of genotypic descriptors of the national pig populations, the evaluationfor which some simple experimental protocols are described in literature. Other lack of knowledge involved relationshipsto predict the anatomical growth and the evolution of quality parameters of carcass and cuts from the chemical compositionof the body. If a correlation between the chemical composition of the pig's body and production targets is confirmed,then a link between the farmer and the industry can be established and the model could be used to identify more convenientfeeding strategies. The conceptual basis and the way to assess nutrient requirements of heavy pigs have beenclearly identified

Plasma sexual steroids and fatty acid analysis of adult Adriatic sturgeon during reproductive season

The reproductive physiology of sturgeons is a topic of constant relevance, due to some peculiarities that characterize the reproductive cycle of these fish and the need to better understand reproduction to improve methods used in aquaculture programs. Female sturgeons need more than a year to complete their roe maturation and their reproduction occurs every 2-5 years according to species and other variables, while captive males could reproduce every years. The aim of the present study was to investigate the concentrations of estradiol 17\u3b2 (E2), testosterone and fatty acids extracted from Adriatic sturgeon plasma collected during the spawning season, in order to evaluate if plasma analysis could provide useful information to sturgeon farmers in broodstock selection. Fifty-six adult captive Adriatic sturgeon from the breeding stock of Parco del Ticino were included in the trial. Blood samples were collected in mid-June from the caudal vasculature and a gonadal biopsy was collected to determine the sex. The stage of maturity of the female ovarian follicles was evaluated by visual inspection of oocyte/ovarian follicle. Stage of maturity in males was not assessed. Plasma was obtained after centrifugation at 3000g for 15 min of heparinized blood. Sex steroids were measured by RIA analysis, and fatty acids were measured by gas chromatography after direct plasma methylation. The E2 concentration ranged from 0.01 to 19.24 ng/ml, while testosterone ranged from 3.84 to 132.60 ng/ml. Testosterone and E2 concentrations were not different between males and non reproductive females , while their concentration was elevated (109 .1 \ub115.3 and 9.7 \ub15.3 ng/ml respectively) only in females that would spawn during the year of sampling. Female sturgeon with immature oocytes had E2 concentrations that were not statistically different from males (0.42\ub10.54 versus 0.07\ub10.08 ng/ml). Plasma fatty acid analysis was not different between sexes. The most representative fatty acid found was oleic acid (18:1 n-9) followed by palmitic acid (16:0) and DHA (22:6 n-3). Finally, the total amount of plasma fatty acid did not differ between sexes or among females with different stages of gonadal maturation. Our study suggest that E2 plasma concentration could be used as marker to identify which females of Adriatic sturgeon are ready to be used as reproducers

Multifactorial causes of chronic mortality in juvenile sturgeon (Huso huso)

This investigation focused on an episode of chronic mortality observed in juvenile Huso huso sturgeons. The examined subjects underwent pathological, microbiological, molecular, and chemical investigations. Grossly severe body shape deformities, epaxial muscle softening, and multifocal ulcerative dermatitis were the main observed findings. The more constant histopathologic findings were moderate to severe rarefaction and disorganization of the lymphohematopoietic lymphoid tissues, myofiber degeneration, atrophy and interstitial edema of skeletal epaxial muscles, and degeneration and atrophy of the gangliar neurons close to the myofibers. Chemical investigations showed a lower selenium concentration in affected animals, suggesting nutritional myopathy. Other manifestations were nephrocalcinosis and splenic vessel wall hyalinosis. Septicemia due to bacteria such as Aeromonas veronii, Shewanella putrefaciens, Citrobacter freundii, Chryseobacterium sp., and pigmented hyphae were found. No major sturgeon viral pathogens were detected by classical methods. Next-generation sequencing (NGS) analysis confirmed the absence of viral pathogens, with the exception of herpesvirus, at the order level; also, the presence of Aeromonas veronii and Shewanella putrefaciens was confirmed at the family level by the metagenomic classification of NGS data. In the absence of a primary yet undetected biological cause, it is supposed that environmental stressors, including nutritional imbalances, may have led to immune system impairment, facilitating the entry of opportunistic bacteria and mycotic hyphae

Gyrodactylus salinae n. sp. (Platyhelminthes: Monogenea) infecting the south European toothcarp Aphanius fasciatus (Valenciennes) (Teleostei, Cyprinodontidae) from a hypersaline environment in Italy

Background: Historically, non-native species of Gambusia (Poeciliidae) have been used to control larval stages of the Asian tiger mosquito, Stegomyia albopicta Reinert, Harbach et Kitching, 2004 throughout Italy. The potential utility of indigenous populations of Aphanius fasciatus (Valenciennes) (Teleostei: Cyprinodontidae) as an appropriate alternative biological control is currently being explored. A sub-sample of ten fish collected from Cervia Saline, Italy (salinity 65 ppt; 30°C) to assess their reproductive capability in captivity, harboured a moderate infection of Gyrodactylus von Nordmann, 1832 (Platyhelminthes, Monogenea). A subsequent morphological and molecular study identified this as being a new species. Results: Gyrodactylus salinae n. sp. is described from the skin, fins and gills of A. fasciatus. Light and scanning electron microscopical (SEM) examination of the opisthaptoral armature and their comparison with all other recorded species suggested morphological similarities to Gyrodactylus rugiensoides Huyse et Volckaert, 2002 from Pomatoschistus minutus (Pallas). Features of the ventral bar, however, permit its discrimination from G. rugiensoides. Sequencing of the nuclear ribosomal DNA internal transcribed spacers 1 and 2 and the 5.8S rRNA gene and a comparison with all species listed in GenBank confirmed they are unique and represent a new species (most similar to Gyrodactylus anguillae Ergens, 1960, 8.3% pair-wise distance based on 5.8S+ITS2). This represents the first species of Gyrodactylus to be described from Aphanius and, to date, has the longest ITS1 (774 bp) sequenced from any Gyrodactylus. Additional sampling of Cervia Saline throughout the year, found G. salinae n. sp. to persist in conditions ranging from 35 ppt and 5°C in December to 65 ppt and 30°C in July, while in captivity a low level of infection was present, even in freshwater conditions (0 ppt). Conclusions: The ability of G. salinae n. sp. to tolerate a wide range of salinities and temperatures shows its potential to readily adapt to several environmental conditions. These findings, together with the fact that A. fasciatus is a protected species and is considered as a biological control organism, necessitate further studies on the ecology and virulence of G. salinae n. sp