The combined effects of temperature and GnRHa treatment on the final stages of sexual maturation in Atlantic salmon (Salmo salar L.) females

Atlantic salmon (Salmo salar L.) females (2 SW), maturing for the first time, were reared under one of three temperature regimes (high: 14.3 ± 0.5°C; natural: 10.6 ± 1.0°C; and cold: 6.9 ± 1.0°C) in combination with one of two experimental treatments; an injection of GnRH analogue (GnRHa) contained in biodegradable microspheres, or a sham injection (microspheres only). The six experimental groups were then reared under simulated natural photoperiod for 4 weeks. Blood samples were drawn for analysis of plasma steroid levels and the fish were inspected for ovulation weekly. Batches of stripped eggs were incubated in triplicate incubators in raceways until the eyed stage. Treatment with GnRHa resulted in a substantial advancement and synchronization of ovulation at all temperatures, while exposure to cold water also appeared to advance ovulation slightly. While 75% (warm and cold) to 90% (natural) of GnRHa fish ovulated during the 4-week trial, only 30% of sham-treated females exposed to cold water, and none of the sham-treated fish held at higher temperatures, ovulated during this period. Survival rates of embryos to the eyed-stage were significantly higher for broodstock exposed to cold water. Plasma levels of testosterone (T), 17β-oestradiol (E₂), and 17α,20β-dihydroxy-4-pregnen-3-one (17,20βP) were all significantly affected by treatment with GnRHa and, to a lesser extent, temperature. The efficiency of GnRHa in counteracting the negative effects of high temperature on ovulation and the associated changes in circulating sex steroids suggest that temperature inhibition operates at least in part at the brain or pituitary.     

The N-acetyl-D-glucosamine specific adhesin from Mannheimia haemolytica activates bovine neutrophils oxidative burst

In this work we identified specific bovine leukocytes that were bound by the Mannheimia haemolytica adhesin molecule (MhA) and the biological effect on the leukocytes. Histochemical staining and flow cytometry showed that MhA bind neutrophils (90%) and monocytes (5%). MhA induced an oxidative response in purified neutrophils; this effect was 1.5-fold higher than the effect observed with control cells activated with Zymosan. Cellular binding by MhA was inhibited with GlcNAc and its oligomers, as well as by glycoproteins containing tri- and tetra-antennary N-glycosydically linked glycans. MhA-induced oxidative burst was significantly inhibited by GlcNAc, iodoacetamide, superoxide dismutase, and piroxicam (p<0.05). Our findings suggest that among bovine leukocytes, neutrophils are the main target for MhA, inducing production of oxidative radicals by non-opsonic mechanism that seem to play an important role in tissue damage during mannheimiosis.     

Lectin from Phaseolus acutifolius var. escumite: chemical characterization, sugar specificity, and effect on human T-lymphocytes

Purification of the lectin from Phaseolus acutifolius var. escumite was achieved by affinity chromatography on a column containing glutaraldehyzed membranes from blood group O erythrocytes. The lectin is a tetrameric glycoprotein of 121 kDa with 10% of sugar by weight composed by four subunits of 30 kDa as determined by SDS-PAGE. The lectin is composed of four isolectins as determined by ion-exchange chromatography on a mono-S column. The lectin and its isolectins showed identical NH2 terminal residues (ANDLSFNFQR FNETN) with homology to the PHA leucoagglutinin-precursor. Peptide mass fingerprint from each lectin isoform determined from tryptic peptides by MALDI-TOF (matrix assisted laser desorption ionization-time-of-flight) showed differences among subunits, thus suggesting microheterogeneity in their amino acid sequences or different glycosylation patterns. The lectin and its four isolectins agglutinated erythrocytes without serological specificity and showed mitogenic activity on human leukocytes; moreover, the main effect was rather toward CD8+ than to CD4+ human peripheral lymphocytes. The lectin from escumite was not inhibitable by simple sugars; however, the specificity of the lectin and its isoforms was mainly addressed toward galactose residues present in bi- or triantennary N-acetyllactosamine-type glycans.     

Peanut and Amaranthus leucocarpus lectins discriminate between memory and naive/quiescent porcine lymphocytes

Lectins are relevant tools to isolate and characterize different cellular sub-populations. In this work, we used the lectins Arachis hypogaea (Peanut agglutinin, PNA) and Amaranthus leucocarpus (ALL), specific for Galss1, 3GalNAc, to characterize naive and memory lymphocytes from pigs, experimentally infected with the porcine rubulavirus (RvP). Our results showed that both lectins recognized preferentially lymphocytes with the CD4(+)CD8(+) phenotype (P<0.05). The phenotypic analysis of the cells recognized by these lectins indicated that PNA(+) lymphocytes showed higher rate of the CD29 antigen (PNA(+)CD29(high)) than ALL(+) (ALL(+)CD29(low)). The number of PNA(+)CD29(high) lymphocytes increased after 8 weeks of experimental infection with RvP, and most of the ALL(+)CD29(low) cells became CD29(middle). PNA(+) lymphocytes isolated from infected pigs proliferated after stimulation with the RvP, whereas ALL(+) cells did not. In vitro assays indicated that the ALL(+) cells from previously infected pigs diminished from 7.5 +/- 2 to 0.5 +/- 0.3% after RvP stimulation; whereas PNA(+) cells increased from 4 +/- 1 to 42 +/- 2%, whereas no modification in ALL(+) or PNA(+) cellular population was identified in lymphocytes from naive animals after RvP stimulation. Our results suggest that the cellular distribution/organization of the O-glycosydically linked glycans on lymphocytes may correlate with biological functions, and that PNA could be a tool to isolate specifically porcine memory T cell subsets, whereas ALL could be useful to isolate naive/quiescent T lymphocytes.     

Induced Spawning of Wild American Shad Alosa sapidissima Using Sustained Administration of Gonadotropin-Releasing Hormone Analog (GnRHa)

American shad Alosa supidissima broodstock were collected from the Susquehanna River during their spawning migration. Mean volume of expressible milt (± standard deviation) was 2.5 (±1.7) mL/kg body weight; mean spermatozoid count was 66.2 ± 10⁹ (±163 ± 10⁹) spermntozoa/mL; and duration of 50% motility was 36.5 (±10.3) see. Ovarian biopsies indicated the presence of oocytes of various sizes (200–2,000 μm in diameter) and stages of development. Fish were implanted with a delivery system loaded with gonadotropin-releasing hormone analog (GnRHa) and started spawning 2 d after treatment. Fertile eggs were collected daily for the next 9 d, for a total of 50,100 eggs/kg body weight with a mean fertilization success of 62%. Upon cessation of spawning, the ovaries of all females still contained large numbers of oocytes at various stages of development, as at the beginning of the experiment, but with a greater number of atretic oacytes. Our observations show that American shad have an asynchronous ovarian development, and treatment with a GnRHa delivery system is effective in inducing several successive spawns of fertile eggs.     

Long photoperiod delayed spawning and increased somatic growth in gilthead seabream (Sparus aurata)

The effects of extended photoperiods, mimicking the longest day of the year, were studied in 1- and 2-year seabream. The photoperiod regimes started in late July, 36 and 39 days after the summer solstice and continued for 11 months, well beyond the natural reproductive season of December–March. Regime 1 (long day, 15.5L:8.5D), which used natural and fluorescent light, reduced the incidence of maturity in both year classes and females did not spawn although some gonadal development was observed. Among all 1-year sampled fish of regime 1, a maximum of 5% became spermiating males (March) and 5% reached the yolk granule stage of vitellogenesis (VO3; 250–400 μm diameter) by May. Among 2-year sampled fish of regime 1, 45% became spermiating males and 25% were females, which reached the advanced vitellogenesis stage (VO4; 400–600 μm) by April. Regime 2 (skeleton photoperiod), consisting of natural light and a 1.5-h pulse of fluorescent light during the period 14–15.5 h after sunrise, postponed gonadal development and spawning for up to 3 months. In this regime, a maximum of 80% of 1-year sampled fish were spermiating males in February and a maximum of 10% were VO3 stage females in March. In the sampled 2-year fish, the maximum levels were 50% spermiating males in February and 25% VO3 stage females in March. Control fish, which were exposed to the natural photoperiod (29°34′N), spawned during their natural season. The maximum levels for 1-year sampled control fish were 95% spermiating males and no females in December, while among 2-year sampled fish, maxima of 75% males in February and 45% VO4 stage females in November. Final average weights of photoperiod treated fish (1-year=430 g—regime 1, 400 g—regime 2; 2-year=582 g—regime 1, 518 g—regime 2) were significantly greater (p<0.05) than control fish (1-year=341 g; 2-year=476 g). Daily feed consumption (g/100 g fish) dropped from an average of 1.83 to 0.93 g for 1-year fish during August–December and from 0.88 to 0.54 g for 2-year fish during the same period. This was correlated with reduced autumn and winter water temperatures (26–20°C summer to winter change) and increased fish weight in all treatments. Daily feed consumption was also affected by the onset of spawning in the control and regime 2 (skeleton photoperiod) treatments of both 1- and 2-year fish. The efficiency of feed utilization (FCR) and protein and energy retention were all affected by the photoperiod regimes and followed the same pattern of decrease as weight gain, regime 1 (long day)>regime 2 (skeleton photoperiod)>control.     

Environmentally sustainable land-based marine aquaculture

Reduced fishery harvests and increased consumer demand for seafood have precipitated an increase in intensive fish farming, predominantly in coastal and open ocean net-pens. However, as currently practiced, aquaculture is widely viewed as detrimental to the environment and typical operations are vulnerable to environmental influences, including pollution and endemic diseases. Here we report the development of a land-based, marine recirculating aquaculture system that is fully contained, with virtually no environmental impact as a result of highly efficient biological waste treatment and water recycling. Over 99% of the water volume was recycled daily by integrating aerobic nitrification to eliminate toxic ammonia and, for the first time, simultaneous, anaerobic denitrification and anaerobic ammonium oxidation, to convert ammonia and nitrate to nitrogen gas. Hydrogen sulfide generated by the separated endogenous organic solids was used as an electron source for nitrate reduction via autotrophic denitrification and the remaining organic solids were converted to methane and carbon dioxide. System viability was validated by growing gilthead seabream (Sparus aurata) from 61 g to 412 g for a total of 1.7 tons in a record 131 days with 99% fish survival. Ammonia nitrite and nitrate did not exceed an average daily concentration of 0.8 mg/l, 0.2 mg/l and 150 mg/l, respectively. Food conversion values were 16% lower than recorded levels for net-pen aquaculture and saltwater usage of less than 16 l/every kg of fish produced. The system is site-independent, biosecure, devoid of environmental contaminants and is not restricted to a single species.     

Sperm Characteristics of Precocious 1-year-old Male Striped Bass Morone saxatilis

Abstract.— Captive-reared, 10-mo-old, male striped bass Morone saxatilis were sampled monthly for testicular development between February and June 1994. One of the five males sampled in February showed precocious testicular development and had a gonado-somatic index (GSI) of 1.26%. while the other four fish had immature testes with a mean GSI ± s^x, of 0.17 ± 0.03%. Spermiating individuals were present from April to June. In April the average body weight (BW) of spermiating males was 65 ± 4 g and their GSI reached a mean value of 4.75 ± 0.52%. In June, milt collected from ten precocious males contained motile spermatozoa with a mean of 31 ± 7% of the sperm showing forward movement. Mean milt volume and sperm concentration were 1.67 ± 0.41 mL/kg BW and 92.3 ± 1.8 ± 10⁹ spermatozoa/mL. respectively. These data show that male striped bass reared in captivity can reach sexual maturity during their first year. This is one year earlier than previously reported for striped bass in mid-Atlantic regions.     

A diluent for sperm cryopreservation of gilthead seabream, Sparus aurata

Sperm of gilthead seabream, Sparus aurata, was diluted with solutions of different osmolarities and pH. The effect of the different diluents on sperm motility (intensity and percentage of motile sperm) was studied. Motility was induced as early as 10 s after mixing the sperm with diluents having an osmotic pressure higher than 500 mOsm/l. The intensity of motility decreased when the osmotic pressure was reduced, and was zero or significantly inhibited when the osmotic pressure of the diluent (300–380 mOsm/l) was close to that of the fish's seminal plasma (364.6±3.03 mOsm/l). The pH of the diluent did not have any effect on sperm motility (in a range of 6.8 to 8.9). A diluent which prevents spermatozoa motility (osmotic pressure 375 mOsm/l and pH 7.35) was successfully used to cryopreserve S. aurata sperm at −196°C. This diluent is considered promising for the long-term preservation of gilthead seabream sperm.