Evaluation of β-(1 → 3, 1 → 6)-glucans and High-M alginate used as immunostimulatory dietary supplement during first feeding and weaning of Atlantic cod (Gadus morhua L.)

Stimulation of the non-specific defence enhances the disease resistance and growth, and has good potentials as a measure for increased microbial control in juvenile production of marine fish and shellfish. So far, the most commonly used immunostimulants are β-(1 → 3, 1 → 6)-glucans, and in this study the stimulatory potential of a β-(1 → 3, 1 → 6)-glucan of marine origin, the storage polysaccharide from the marine diatom Chaetoceros mülleri, was examined. The glucan (chrysolaminaran) was extracted from cultures of C. mülleri, and used as a dietary supplement in two first feeding experiments with larvae of Atlantic cod Gadus morhua L. In one experiment the microalgal glucan was compared to the commercial yeast-glucan product MacroGard®, and in the other to an alginate with a high content of mannuronic acid (High-M alginate) isolated from Durvillaea antarctica. The stimulants were given via rotifers, and weaning to formulated feed was initiated at day 17 or 18 after hatching. The survival ± SEM at day 27 after hatching was 24.5 ± 2.0%, 14.8 ± 4.5% and 13.1 ± 1.4% for the groups fed C. mülleri-glucan, yeast glucan and for the control, respectively, in the first experiment. The group fed C. mülleri-glucan group had higher survival compared to the control (P < 0.05) group, whereas the yeast glucan had no positive effect on the survival (p > 0.05). The dry weights of the groups at day 27 were low, with 203.2 ± 52.2, 165.2 ± 43.4 and 198.5 ± 58.1 μg per larva for the C. mülleri-glucan, yeast glucan and control groups, respectively. In the second experiment the survival in the period of feeding formulated feed (days 18-30) were 44.6 ± 4.3%, 44.7 ± 1.3%, and 33.8 ± 4.1% survival for the C. mülleri-glucan, High-M alginate and control group, respectively. The cod larvae fed C. mülleri-glucan reached an average weight of 531.6 ± 17.2 μg at day 30, which was significantly higher (p < 0.05) than the control group that had an average of 473.6 ± 3.5 μg. The larvae fed High-M alginate had an average weight of 470.3 ± 31.6 μg per larva at day 30, and not significantly different from the control (p > 0.05). The early weaning to formulated diet had detrimental effect on the growth of the larvae. In both experiments the C. mülleri-glucan group was the only group showing a positive growth rate in the period of weaning to dry feed. The microbial conditions in larval gut and water were monitored with respect to total colony forming units on Marine agar, and Vibrio- and Pseudomonas-like species on selective agars (TCBS and marine Pseudomonas Agar with CFC-supplement). The larvae were rapidly colonised after hatching, but no or weak effects of the stimulants were observed on the colonisation rates or the composition. The total CFU varied from 10¹ to 10² CFU per μg larva after initiation of the first feeding. The percentages of Pseudomonas-like bacteria increased throughout the period, whereas the levels of Vibrio-like bacteria were low and stable. The chrysolaminaran from the diatom C. mülleri was shown to be a promising candidate for use as an immunostimulatory feed additive, and which should be further explored.     

Influence of storage conditions on viability of quiescent copepod eggs (Acartia tonsa Dana): effects of temperature, salinity and anoxia

Copepods have proven to be an ideal source of live food for the production of marine fish larvae in aquaculture. Therefore, there is a need to develop new methods for production and storage of copepod eggs that can be hatched and used at fish farms. In the present study quiescent eggs of Acartia tonsa were stored for periods up to 35 weeks at different temperatures, salinities and oxygen conditions in a full factorial experiment. None of these storage conditions seemed to induce diapause in eggs even though this has been reported by other authors. The most promising storage conditions were those involving low temperature (<5°C), medium salinity (10–20 ppt) and anoxia. The practical aspects of these results for aquaculture are discussed.     

Interacting effects of diet and environmental temperature on biochemical parameters in the liver ofLeuciscus idus melanotus (Cyprinidae: Teleostei)

One year old golden ide (Leuciscus idus melanotus) were starved for 2 weeks at 20°C (time zero) and then they were fed either freeze-dried mosquito larvae (natural diet) or a commercial fish chow (artificial diet) at an ambient temperature of 14°C and 20°C, respectively. Growth parameters and biochemical data in the liver were measured at time zero and after 3 and 7 weeks of specific regimen. If compared to natural food, the artificial diet caused transient increase in anabolic activity of the liver, but prevented long-lasting hepatocyte proliferation (increase of total liver DNA). After 3 weeks on artificial diet, the body weight was significantly higher and the liver mass doubled compared to ide kept on the natural regimen; tissue DNA content indicated that the hepatocyte volume increased mainly by increased protein content and corresponding uptake of water; lipid and glycogen contents were increased by a factor of 3–6, and the RNADNA ratio was increased accordingly. The growth of the ide, as reflected by the condition factor and hepatosomatic index (HSI), was virtually stopped when the artificial regimen was maintained for 7 weeks, whereas on natural food the condition factor was increased and liver weight and DNA were doubled. Growth of the fish maintained at 14°C was significantly lower than that of fish maintained at 20°C, though the total liver DNA was still increased on the natural regimen during the experiment. Less lipid was stored by fish maintained at 14°C, whereas protein and glycogen deposits were enlarged, if compared to fish maintained at 20°C. The specific activity of cytochrome oxidase in liver mitochondria and of NADPH cytochrome c reductase in the microsomal fraction were found independent of diet and ambient temperature. Oxidative capacity of hepatocytes (mitochondrial protein/mg DNA) remained unchanged, and microsomal protein/cell appeared reduced in response to the artificial food. However, reduction of cell number on this diet resulted in less total mitochondrial and microsomal protein in the organ. Golden ide liver is found to be a sensitive experimental model to characterize the mutual influence of diet and temperature on fish; the results are discussed with regard to the usage of golden ide as test fish for water pollution.     

Lower prevalence of fin erosion in mature than in immature Atlantic salmon (Salmo salar) parr

Samples of hatchery-reared Atlantic salmon parr (age 0+ and 1+) from two different Norwegian river strains were investigated for connections between sexual maturity and prevalence of dorsal fin ray erosion. Mature males showed consistently lower prevalence as well as better recovery than immature males. Among the immature fish, no differences were detected between males and females. The results are discussed relative to morphological and physiological changes in maturing salmon parr.     

Effects of egg disinfection and incubation temperature on early life stages of spotted wolffish

Eggs of spotted wolffish (Anarhichas minor Olafsen)were incubated at constant 4, 6 and 8 °C, and disinfected withglutaric dialdehyde (150 p.p.m. for 5 min) once ortwice a month during two thirds of the incubation period, to prevent growth ofmicroorganisms. Hatching of apparently normal larvae started earlier when eggswere disinfected twice a month compared to once a month at all incubationtemperature regimes. The time to 50% hatch was 900 and 920 day-degrees (16 and16,5 weeks) at 8 °C, 835 and 880 day-degrees (20 and 21 weeks)at 6 °C and 725 and 800 day-degrees (26 and 28,5 weeks) at 4°C, in the egg groups disinfected twice or once a month,respectively. The best survival until hatching was noted when eggs weredisinfected twice a month and incubated at 6 and 8 °C.Survivalwas very low at 4 °C. Prematurely hatched larvae wereregistered in all egg groups disinfected twice a month and the highestfrequencywas noted in the 8 °C groups. The larval weight at normalhatching in the 6 and 8 °C groups was negatively correlatedwith incubation temperature and intervals of disinfection during the incubationperiod, but after 42 days feeding with live feed (unenrichedArtemia) the weights of the larvae were not significantlydifferent. The specific growth rates of the larvae from the eggs incubated at 6°C and 8 °C were 3.0% and 3.2%, respectively.The mean survival of larvae was between 88% and 96% at 42 days post-hatching.Young wolffish originating from the 6 °C incubation groupsshowed lowest mortality.     

Marennine,Promising Blue Pigments from a Widespread Haslea Diatom Species Complex

In diatoms, the main photosynthetic pigments are chlorophylls a and c, fucoxanthin, diadinoxanthin and diatoxanthin. The marine pennate diatom Haslea ostrearia has long been known for producing, in addition to these generic pigments, a water-soluble blue pigment, marennine. This pigment, responsible for the greening of oysters in western France, presents different biological activities: allelopathic, antioxidant, antibacterial, antiviral, and growth-inhibiting. A method to extract and purify marennine has been developed, but its chemical structure could hitherto not be resolved. For decades, H. ostrearia was the only organism known to produce marennine, and can be found worldwide. Our knowledge about H. ostrearia-like diatom biodiversity has recently been extended with the discovery of several new species of blue diatoms, the recently described H. karadagensis, H. silbo sp. inedit. and H. provincialis sp. inedit. These blue diatoms produce different marennine-like pigments, which belong to the same chemical family and present similar biological activities. Aside from being a potential source of natural blue pigments, H. ostrearia-like diatoms thus present a commercial potential for aquaculture, cosmetics, food and health industries.     

Ash and biochar amendment of coarse sandy soil for growing crops under drought conditions

Appropriate soil amendments may increase plant available water and crop yields on coarse sandy soils under drought conditions. In this study, we applied straw ash or straw biochar from gasification to a Danish coarse sandy subsoil to assess the effects on soil water retention, evapotranspiration and crop yields. Spring barley (2016, 2017) and winter wheat (2018) were grown over three years in columns containing 25cm of organic matter-rich topsoil, 80 cm of amended coarse sandy soil (1.5%, 3%, 6% wt. ash or 1% wt. biochar or control soil) and 45 cm of un-amended subsoil. Precipitation, evaporative demands and soil moisture were recorded across the growth seasons, with 2018 having severe drought conditions. This year evapotranspiration levels increased with increasing ash and biochar content (by 54% and 33% for the 6% ash- and 1% biochar-amended soils, respectively), and plant dry matter increased by 18% in both the 1% biochar- and 6% ash-treated soils compared to the untreated control. A linear relationship was established between in situ field capacity and ash dosage (R² = .96), showing an increase of 2.2% per percentage (wt.) of ash added, while the 1% biochar treatment increased the capacity by 3.5%, indicating a higher efficiency than for ash. However, we did not find significant positive effects on grain yields. The results show that ash and biochar have the potential to significantly increase soil water retention, evapotranspiration and total dry matter yield in drought conditions, but that this may not correspond to an increase in grain yield.     

Mites of the genus Carabodes (Acari,Oribatida) in Norwegian coniferous forests: occurrence in different soils,vegetation types and polypore hosts

Oribatid mites (Acari) represent a considerable part of the biodiversity in Fennoscandian forests, but our knowledge about their habitat requirements is limited. We studied 10 Carabodes species in the forest floor of seven coniferous forest types, and in dead fruiting bodies (sporocarps) of 6 species of wood-living polypore fungi in southern Norway. The most common Carabodes species in soil were rare in sporocarps, and vice versa. The density of several ground-living species was significantly influenced by vegetation type and soil type. Carabodes willmanni and C. subarcticus were considered as lichen feeders on the ground, and occurred abundantly in Cladonia-rich pine forests. Three species, C. femoralis, C. areolatus and C. reticulatus, seem to be sporocarp specialists. Their relative numbers were rather similar in dead sporocarps of five different fungal species, including annual and perennial sporocarps, soft and hard. This was in contrast to beetles from the same sporocarps, which in a previous study proved to be strongly host-specific. Although being tolerant to different fungal species, the association of certain Carabodes species to dead sporocarps could make them vulnerable in forests with little dead wood and few sporocarps.     

Investigating the mechanisms for the opposing pH relationships of fungal and bacterial growth in soil

Soil pH is one of the most influential variables in soil, and is a powerful factor in influencing the size, activity and community structure of the soil microbial community. It was previously shown in a century old artificial pH gradient in an arable soil (pH 4.0-8.3) that bacterial growth is positively related to pH, while fungal growth increases with decreasing pH. In an attempt to elucidate some of the mechanisms for this, plant material that especially promotes fungal growth (straw) or bacterial growth (alfalfa) was added to soil samples of the pH gradient in 5-day laboratory incubation experiments. Also, bacterial growth was specifically inhibited by applying a selective bacterial growth inhibitor (bronopol) along the entire pH gradient to investigate if competitive interaction caused the shift in the decomposer community along the gradient. Straw benefited fungal growth relatively more than bacterial, and vice versa for alfalfa. The general pattern of a shift in fungal:bacterial growth with pH was, however, unaffected by substrate additions, indicating that lack of a suitable substrate was not the cause of the pH effect on the microbial community. In response to the bacterial growth inhibition by bronopol, there was stimulation of fungal growth up to pH 7, but not beyond, both for alfalfa and straw addition. However, the accumulation of ergosterol (an indicator of fungal biomass) during the incubation period after adding alfalfa increased at all pHs, indicating that fungal growth had been high at some time during the 5-day incubation following joint addition of alfalfa and bronopol. This was corroborated in a time-series experiment. In conclusion, the low fungal growth at high pH in an arable soil was caused to a large extent by bacterial competition, and not substrate limitation.