Transport of juvenile dusky grouper Epinephelus marginatus under different packing densities: Metabolic and haematological responses

The aim of this study was to investigate a suitable packing density for the transport of juvenile dusky grouper, Epinephelus marginatus, based on the evaluation of stress responses in blood. After acclimation, fish were placed in plastic bags and transported for 8 hr on paved road at densities of 28, 45 and 64 g/L. Water quality was monitored before and after transport. Blood was collected before, upon arrival (0 hr), after 2 and 24 hr of transport. Plasma cortisol, blood glucose, partial pressures of O₂ (pO₂) and CO₂ (pCO₂), blood pH and HCO₃^? were evaluated. Blood smears were prepared for the verification of leucocyte profile and neutrophils:lymphocyte ratio (N:L). Blood pCO₂, pH and HCO₃^? increased significantly after transport for all treatments compared with pre‐transport. Glucose levels increased at the higher density whereas no effects were observed on plasma cortisol and pO₂ levels. Upon arrival, all treatments showed lymphopenia and neutrophilia which increased N:L ratio. Although lymphopenia was observed in higher densities until 2 hr after transport, haematological parameters were fully restored within 24 hr post transport. Furthermore, no mortalities were observed throughout the experimental period. Based on the transient physiological changes observed in this study, juvenile dusky grouper can be safely transported in plastic bags for 8 hr at a density of up to 64 g/L.     

Mycorrhizal activity and diversity in a long-term organic Mediterranean agroecosystem

In organic agriculture, soil fertility and productivity rely on biological processes carried out by soil microbes, which represent the key elements of agroecosystem functioning. Arbuscular mycorrhizal fungi (AMF), fundamental microorganisms for soil fertility, plant nutrition and health, may play an important role in organic agriculture by compensating for the reduced use of fertilizers and pesticides. Though, AMF activity and diversity following conversion from conventional to organic farming are poorly investigated. Here we studied AMF abundance, diversity and activity in short- and long-term organically and conventionally managed Mediterranean arable agroecosystems. Our results show that both AMF population activity, as assessed by the mycorrhizal inoculum potential (MIP) assay, the percentage of colonized root length of the field crop (maize) and glomalin-related soil protein (GRSP) content were higher in organically managed fields and increased with time since transition to organic farming. Here, we showed an increase of GRSP content in arable organic systems and a strong correlation with soil MIP values. The analysis of AMF spores showed differences among communities of the three microagroecosystems in terms of species richness and composition as suggested by a multivariate analysis. All our data indicate that AMF respond positively to the transition to organic farming by a progressive enhancement of their activity that seems independent from the species richness of the AMF communities. Our study contributes to the understanding of the effects of agricultural managements on AMF, which represent a promising tool for the implementation of sustainable agriculture.     

Phyto-dehydration of confined sludge: a sustainable approach for the management of polluted ponds

Purpose

Leaf transpiration drives many of the processes involved in phyto-technologies, and it can represent a useful mechanism to remove water from different kind of storage basins presenting inorganic, organic or microbiological contamination (phyto-dehydration), with the aim to reduce the risk of environmental contamination. In this framework, a mesocosm-scale trial was carried out to test the capacity of different helophyte species to reduce the excess of water in an artificial pond filled with oversaturated sludge.

Materials and methods

The sludge derives from the digestion of pig slurries, presents high levels of zinc and copper and for most of the year is covered by a water layer of about 20 cm due to rainfalls. This layout (water layer over the sludge) was reproduced inside the mesocosms, where four helophyte species (Phragmites australis and a mix of Carex acutiformis, Iris pseudacorus and Juncus effusus) were planted on floating frames. Plant growth and functionality were monitored for one year, along with their water consumption capacity; the vegetation impact on sludge chemistry, sludge microbial community and sludge greenhouse gases emission/uptake were also evaluated. The sensitivity of the phyto-dehydration system to the reduction of water level occurring during summer in the pond was investigated reducing the water input to the mesocosms.

Results and discussion

P. australis and C. acutiformis successfully established in the mesocosms, while a significant mortality was recorded for I. psudacorus and J. effusus. Once established, plants were able to grow in the mesocosms, and no metal toxicity effect was observed on photosynthesis rates that were comparable with values reported for natural stands of the species. Plants significantly increased (from 24 to 63%, depending on the species) the amount of water lost by the mesocosms and counteracted the reduction of sludge organic carbon that could lead to a mobilization of the heavy metals bound to organic matter: furthermore, plants decreased the rates of mesocosm greenhouse gas emission and reduced the sludge pathogen (Enterobacteriaceae) occurrence. Water limitations reversibly reduced the water consumption and CO₂ uptake capacity of the mesocosms.

Conclusions

The results of this study demonstrated that the water balance of a sludge/water system can be effectively modified through the phyto-dehydration approach, increasing significantly the amount of water lost. Although the low tolerance of two species to the sludge/water environment after plantation needs to be further investigated, this phyto-technology can represent a promising approach to manage the excess of water in polluted ponds.

     

Morphological characterization of the adherence and invasion of Streptococcus agalactiae to the intestinal mucosa of tilapia Oreochromis sp.: An in vitro model

Streptococcosis in tilapia Oreochromis sp. is possibly the most important bacterial disease for fish production worldwide. In Colombia, streptococcosis is caused by Streptococcus agalactiae (GBS), but in other countries, Streptococcus iniae is also involved. Prevention of streptococcosis is required and must be addressed for economic, social, international trade and public health reasons. This research used an in vitro culture of tilapia intestine to detail the intestinal mucosal response once the pathogen contacts the epithelium. We show that S. agalactiae sheds off its capsule to adhere to the epithelium. The bacterium adheres as a single individuum, in groups or in chains and is able to divide on the apical border of enterocytes. GBS adheres at and invades exclusively through the apical portion of the intestinal folds, using the transepithelial route. Once within the cytoplasm of enterocytes, the bacteria continue to divide. On the basolateral side of the epithelium, the microorganisms leave the cells to reach the propria and travel through the microcirculation. No evidence of an immuno‐inflammatory reaction or goblet cell response in the epithelium or the lamina propria was seen during the process of adherence and invasion of the pathogen.     

Alginate Hydrogels Coated with Chitosan for Wound Dressing

In this work, a coating of chitosan onto alginate hydrogels was realized using the water-soluble hydrochloride form of chitosan (CH-Cl), with the dual purpose of imparting antibacterial activity and delaying the release of hydrophilic molecules from the alginate matrix. Alginate hydrogels with different calcium contents were prepared by the internal setting method and coated by immersion in a CH-Cl solution. Structural analysis by cryo-scanning electron microscopy was carried out to highlight morphological alterations due to the coating layer. Tests in vitro with human mesenchymal stromal cells (MSC) were assessed to check the absence of toxicity of CH-Cl. Swelling, stability in physiological solution and release characteristics using rhodamine B as the hydrophilic model drug were compared to those of relative uncoated hydrogels. Finally, antibacterial activity against Escherichia coli was tested. Results show that alginate hydrogels coated with chitosan hydrochloride described here can be proposed as a novel medicated dressing by associating intrinsic antimicrobial activity with improved sustained release characteristics.     

Optimizing initial feeding of the Pike silverside Chirostoma estor: oil droplet depletion,point of no return,growth and fatty acid utilization in larvae fed enriched rotifers

The point of no return (PNR) and disappearance of the oil droplet were measured in Chirostoma estor larvae as a function of the time of first feeding. In a separate trial, growth and survival of larvae fed rotifers enriched with Chlorella sp., cod liver oil and corn oil were assessed. Fatty acid and lipid composition of eggs, oil droplets, egg yolk, feed and larvae were also evaluated. The PNR was found between 7 and 8 days posthatching (dph). Total oil droplet depletion occurred between 7 and 11 dph, depending on the time of first feeding. Best growth and survival were obtained in larvae fed with Chlorella‐enriched rotifers, followed by those fed cod liver oil‐enriched rotifers. In larvae fed corn oil, Chlorella and cod liver oil‐enriched rotifers, total oil droplet depletion took place on days 9, 10 and 11, respectively. There was a direct relationship between presence and duration of oil droplets and the survival of larvae under different starvation conditions. The feed source could prolong the existence of the oil droplet depending on particular dietary supply of essential fatty acids; the time of its disappearance could be a useful indicator of larval vigour and health status.     

Introgressive Hybridization in Potato Revealed by Novel Cytogenetic and Genomic Technologies

Potato is the third most important food crop in the world and is crucial to ensure food security. However, increasing biotic and abiotic stresses jeopardize its stable production. Fortunately, breeders count on a rich pool of wild relatives that provide sources for disease resistance and tolerance to environmental stresses. To use such traits effectively, breeders require tools that facilitate exploration and exploitation of the genetic diversity of potato wild relatives. Introgression programs to incorporate such alien chromatin into the crop have so far relied on cytogenetic and genetic studies to tap desired traits from these wild resources. The available genetic and cytogenetic tools, supplemented with more recent genomic technologies, can assist in the use of potato relatives in pre-breeding. This information can also facilitate cisgenesis and genome editing to improve potato cultivars. Despite the abundant and rapidly growing genomic information of potato, that of its wild relatives is still limited.     

Artificial reproduction protocol,from spawning to metamorphosis,through noninvasive methods in Patella caerulea Linnaeus, 1758

Controlled reproduction is a requirement for developing effective mollusc cultivation for commercial or restoration purposes. In this study, a protocol for spawning induction using noninvasive methods in limpets was developed, using the common Mediterranean species, Patella caerulea Linnaeus, 1758. Six nonlethal spawning induction treatments were tested: three chemical (two concentrations of H₂O₂ and KCl) and three physical (bubbling, warm and cold thermal shock). All treatments, except thermal shocks, induced the spawning of fertile gametes. Bubbling resulted the best treatment in providing spawning response, being the easiest and least invasive method tested. After eggs fertilization, larval development was followed until metamorphosis, testing fed and unfed conditions. Settlement took place after 7 days. The developed protocol represents a benchmark for further application to other limpets, for aquaculture or repopulation.