Implication of mucus-secreting cells,acidophilic granulocytes and monocytes/macrophages in the resolution of skin inflammation caused by subcutaneous injection of λ/κ-carrageenin to gilthead seabream (Sparus aurata) specimens

To date, the mechanisms of inflammation have been poorly studied in fish of commercial interest, due to the lack of development of appropriate experimental models. The current study evaluated a local inflammation triggered by a polymeric carrageenin mixture (a mucopolysaccharide derived from the red seaweed Chondrus crispus) in the skin of gilthead seabream (Sparus aurata). Fish were injected subcutaneously with phosphate-buffered saline (as control) or λ/κ-carrageenin (1%), and skin samples from the injection sites were collected 1.5, 3 and 6 hr post-injection, processed for inclusion in paraplast and stained with haematoxylin–eosin, Alcian blue or periodic acid–Schiff. Furthermore, immunohistochemistry and expression analyses of several cells’ markers and proinflammatory genes were also analysed in samples of the injected sites. Microscopic results indicated an increased number of skin mucus-secreting cells and acidophilic granulocytes in the skin of fish studied at 1.5 hr and 3 hr post-injection with carrageenin, respectively, with respect to the data obtained in control fish. Otherwise, both the gene expression of the non-specific cytotoxic cell marker (granzyme B, grb) and the proinflammatory cytokine (interleukin-1β, il-1β) were up-regulated at 1.5 hr in the skin of fish injected with carrageenin compared with the control fish, whilst the gene expression of acidophilic granulocyte markers (NADPH oxidase subunit Phox22 and Phox40, phox22 and phox40) was up-regulated at 3 and 6 hr in the carrageenin group, compared with the control group. In addition, the gene expression of myeloperoxidase (mpo) was also up-regulated at 6 hr in the skin of fish injected with carrageenin in comparison with control samples. The present results indicate the chronological participation of two important immune cells involved in the resolution of the inflammation in the skin of gilthead seabream.     

Different mycorrhizal fungal strains determine plant community response to nitrogen and water availability

Most research on the mycorrhizal positive–negative responsiveness continuum (or “mutualism–parasitism continuum”) has focused on individual plant species growing at different levels of P availability. Here, we explore this continuum in an experimental plant community inoculated with three arbuscular mycorrhizal (AM) fungal strains (both single and mixed) growing under four resource availability scenarios. These scenarios are a factorial combination of two levels of water and N availability. Each AM fungal strain had a different origin: an arid ecosystem, a farmland, and a mine. We hypothesized that the response of the plant community to mycorrhizal inoculum would depend on the associated AM fungal strain and would be negatively related with increased nitrogen and water availability. Our results showed that mixed‐strain AM fungal inoculation had more positive effects along a wider range of water and N availability scenarios than single‐strain inoculation. In contrast, mycorrhizal growth response of plants inoculated with a single AM fungal strain shifted from positive to neutral and negative depending on resource availability. Adaptation of each strain to its local conditions might confer different properties to the mycorrhizal symbiosis. Therefore, we conclude that AM fungal origin and environmental limiting resources are crucial factors to predict plant community mycorrhizal growth response in changing ecosystems.     

Characterization of Industrial Onion Wastes (<Emphasis Type="BoldItalic">Allium cepa</Emphasis> L.): Dietary Fibre and Bioactive Compounds

The food industry produces a large amount of onion wastes, making it necessary to search for possible ways for their utilization. One way could be to use these onion wastes as a natural source of high-value functional ingredients, since onion are rich in several groups of compounds, which have perceived benefits to human health. The objective of this work is to gain knowledge of any differences between the different onion wastes obtained from industry and non-commercial bulbs to use them as food ingredients rich in specific compounds. The results showed that brown skin and top–bottom could be potentially used as functional ingredient rich in dietary fibre, mainly in insoluble fraction, and in total phenolics and flavonoids, with high antioxidant activity. Moreover, brown skin showed a high concentration of quercetin aglycone and calcium, and top–bottom showed high concentration of minerals. Outer scales could be used as source of flavonols, with good antioxidant activity and content of dietary fibre. However, inner scales could be an interesting source of fructans and alk(en)yl cystein sulphoxides. In addition, discarded onions (cvs Recas and Figueres) could be used as a good source of dietary fibre, and cv Recas also as a source of phenolics compounds.     

Prevalence of Francisella noatunensis subsp. orientalis in Cultured Tilapia on the Island of Oahu,Hawaii

Abstract

Francisellosis is an emergent disease in cultured and wild aquatic animals. The causative agent, Francisella noatunensis subsp. orientalis (Fno), is a gram-negative bacterium recognized as one of the most virulent pathogens of warmwater fish. The main objective of this project was to investigate the prevalence of Fno in cultured tilapia (specifically, Mozambique Tilapia Oreochromis mossambicus, Koilapia [also known as Wami Tilapia] O. hornorum, Blue Tilapia O. aureus, and Nile Tilapia O. niloticus hybrids) on the island of Oahu, Hawaii, using conventional and real-time PCR assays followed by statistical modeling to compare the different diagnostic methods and identify potential risk factors. During 2010 and 2012, 827 fish were collected from different geographical locations throughout the island of Oahu. Upon collection of fish, the water temperature in the rearing system and the length of individual fish were measured. Extraction of DNA from different tissues collected aseptically during necropsy served as a template for molecular diagnosis. High correlation between both molecular methods was observed. Moreover, the bacterium was isolated from infected tilapia on selective media and confirmed to be Fno utilizing a species-specific Taqman-based real-time PCR assay. Although a direct comparison of the prevalence of Fno between the different geographical areas was not possible, the results indicate a high prevalence of Fno DNA in cultured tilapia throughout the farm sites located on Oahu. Of the different tilapia species and hybrids currently cultured in Hawaii, Mozambique Tilapia were more susceptible to infection than Koilapia. Water temperature in the rearing systems and fish size also had a strong effect on the predicted level of infection, with fish held at lower temperatures and smaller fish being more susceptible to piscine francisellosis.

Received November 30, 2012; accepted February 22, 2013     

Effects of salinity on various growth stages of muskmelon cv. Revigal

Salt tolerance of muskmelon cv. Revigal was assessed during germination, seedling growth and growth in hydroponic culture. First, germination tests were conducted using four salinity treatments (0, 390, 780 and 1560 mS irf'). At the second stage, seedling growth was studied at salinities of 225,550,775 and 1000 mS m“1. Finally, the influence of salt on the size and organoleptic characteristics of fruit and on the content of various elements in the leaves was studied at salinity levels of 250,500 and 750 mS m'1. Abilities of the seed to germinate and of the seedling to grow adequately were determined under saline conditions of up to 780 mS m_l and 550 mS m“1, respectively. During the last stage, a slight decrement in yield was observed at 500 mS nT1 and fruit mean weight reached 82% of the control. Results suggest that this cultivar is moderately salt-tolerant and can be grown, in principle, at salinities of up to 500 mS m‘. An interesting relationship has been established between the influence of salinity on leaf area of the seedling and on the yield obtained during hydroponic culture; we are therefore proposing seedling leaf area as a major indicator of salt tolerance.     

Mycorrhizal synthesis between Pisolithus arhizus and adult clones of Arbutus unedo in vitro and in nursery

Arbutoid mycorrhizae were synthesized between adult se-lected clones of Arbutus unedo L. and Pisolithus arhizus. Two micro-propagated clones were tested:AL1, in vitro and C1 (acclimatized plants) in nu...     

Debaryomyces hansenii L2‐enriched diet enhances the immunity status,gene expression and intestine functionality in gilthead seabream (Sparus aurata L.)

This study evaluates the effects of the dietary administration of the live yeast Debaryomyces hansenii strain L2 on the immune responses of gilthead seabream for 4 weeks. Cellular immune parameters were measured from serum and head‐kidney leucocytes respectively. The expression levels of immune‐associated genes were quantified by real‐time polymerase chain reaction. In addition, the profile of intestinal microbiota was studied by denaturing gradient gel electrophoresis. The results showed that seabream fed a diet containing D. hansenii had significantly increased cellular immune parameters. The yeast‐supplemented diet up‐regulated the expression of most seabream genes at week 2 and down‐regulated all of them at week 4, except in the head‐kidney. Finally, a reduction in the diversity of the intestinal microbiota was detected in those specimens receiving the yeast‐supplemented diet. These results support the idea that the live yeast D. hansenii strain L2 stimulates the immune system of gilthead seabream.     

Enrichment of gilthead seabream (Sparus aurata L.) diet with microalgae: effects on the immune system

The present study assessed the effects of three orally administered microalgae (Nannochloropsis gaditana, Tetraselmis chuii and Phaeodactylum tricornutum) on different immune parameters and immune-related gene expression of gilthead seabream (Sparus aurata L.). Fish were fed a control or one of six experimental diets and sampled at 2 and 4?weeks of treatment. At the end of the trial, growth performance and different systemic and local immune activities were measured (natural haemolytic complement activity, IgM levels, phagocytosis and respiratory burst). The expression levels of different immune-associated genes (EF-1α, IgM_H, TCR-β, MHCIα, MHCIIα, CSF-1R and β-defensin) were analysed in head-kidney and gut. Administration of N. gaditana and T. chuii resulted in a significant increase in haemolytic complement activity, phagocytic capacity, as well expression level of β-defensin, as well as MHCIIα and CSF-1R, respectively. The P. tricornutum-supplemented diet provoked immunostimulation, and very little effect on gene expression was observed. These results suggest that dietary microalgae enhance gilthead seabream defence activity, which could be very important in order to consider such microalgae as a possible additive in fish diets.     

Use of the probiotic Shewanella putrefaciens Pdp11 on the culture of Senegalese sole (Solea senegalensis, Kaup 1858) and gilthead seabream (Sparus aurata L.)

The application of probiotics on aquatic animals is increasing for a better fish welfare status as well as an environment-friendly activity which are actual demands of modern aquaculture industry. A bacterium from skin mucus of healthy gilthead seabream (Sparus aurata L.) has been isolated and identified as Shewanella putrefaciens Pdp11. Different studies have been done to know its application as probiotic in the Senegalese sole and gilthead seabream farming. This article reviews the studies carried out with this probiotic microorganism focusing on the current knowledge of its in vitro and in vivo mechanisms of action. The results suggested that the probiotic S. putrefaciens Pdp11, due to its beneficial effects, could be used in the aquaculture activity of both species.     

Differential effects of spot blotch on photosynthesis and grain yield in two barley cultivars

Barley spot blotch (SB), caused by Cochliobolus sativus, is an important barley disease which causes extensive grain yield losses. These losses may not always correlate directly with the amount of diseased leaf area. Two barley cultivars, Quebracho (susceptible to SB) and Carumbé (with intermediate susceptibility to SB), were compared in field experiments in 2003, 2004 and 2006. Plots of each cultivar were either inoculated with C. sativus or protected with fungicide under field conditions to generate contrasting treatments: i) diseased, and ii) free of disease, respectively. SB severity over the growing season, photosynthetic rate on leaves with no visible symptoms and grain yield were assessed for each treatment and year. There was no treatment effect on cv. Carumbé, while cv. Quebracho showed a significant yield reduction, even though SB severity during the grain filling period was <10 %. This yield reduction was associated with a reduced photosynthetic rate at the beginning of the grain filling period in cv. Quebracho. A similar experiment was conducted under greenhouse conditions, adding a treatment without inoculum or fungicide. There were no differences in photosynthetic rate or grain yield per plant among treatments. These results suggest a distinct physiological response to SB infection among cultivars affecting leaf photosynthetic rate, and SB severity may not be the best estimator of yield losses caused by SB.