Haematology reference values for Dicentrarchus labrax and Sparus aurata: A systematic review and meta-analysis.

Haematological parameters are frequently used as physiological indicators in aquaculture studies. These parameters also have extended applications in clinical evaluation, diagnosis and prognosis in fish health status. However, no normal reference range of values has been demonstrated in depth for any of these haematological parameters for the European sea bass (Dicentrarchus labrax) or gilthead seabream (Sparus aurata). The main objective of this article is to present for the first time through extended literature review, the haematological parameters normal range values for the two most important aquaculture fish species farmed in Mediterranean Sea, D. labrax and S. aurata, and to demonstrate their similarities and their differences. In this article, we also discuss the environmental and external factors affecting their normal blood parameters values and we propose fundamental guidelines on the reporting units.

Freshwater-adapted sea bass Dicentrarchus labrax feeding frequency impact in a lettuce Lactuca sativa aquaponics system.

The aim of this study is to investigate the effect of three daily fish feeding frequencies, two, four and eight times per day (FF2, FF4, and FF8, respectively) on growth performance of sea bass (Dicentrarchus labrax)and lettuce plants (Lactuca sativa) reared in aquaponics. 171 juvenile sea bass with an average body weight of 6.80 ± 0.095 g were used, together with 24 lettuce plants with an average initial height of 11.78 ± 0.074 cm over a 45-day trial period. FF2 fish group showed a significantly lower final weight, weight gain and specific growth rate than the FF4 and FF8 groups. Voluntary feed intake was similar for all the three feeding frequencies treatmens (p > 0.05). No plant mortality was observed during the 45-day study period. All three aquaponic systems resulted in a similar leaf fresh weight and fresh and dry aerial biomass. The results of the present study showed that the FF4 or FF8 feeding frequency contributes to the more efficient utilization of nutrients for better growth of sea bass adapted to fresh water while successfully supporting plant growth to a marketable biomass.

Bacterial biofilm development during experimental degradation of Melicertus kerathurus exoskeleton in seawater.

Chitinolytic bacteria are widespread in marine and terrestrial environment, and this is rather a reflection of their principle growth substrate's ubiquity, chitin, in our planet. In this paper, we investigated the development of naturally occurring bacterial biofilms on the exoskeleton of the shrimp Melicertus kerathurus during its degradation in sea water. During a 12-day experiment with exoskeleton fragments in batch cultures containing only sea water as the growth medium at 18 °C in darkness, we analysed the formation and succession of biofilms by scanning electron microscopy and 16S rRNA gene diversity by next generation sequencing. Bacteria belonging to the γ- and α-Proteobacteria and Bacteroidetes showed marked (less or more than 10%) changes in their relative abundance from the beginning of the experiment. These bacterial taxa related to known chitinolytic bacteria were the Pseudolateromonas porphyrae, Halomonas aquamarina, Reinekea aestuarii, Colwellia asteriadis and Vibrio crassostreae. These bacteria could be considered as appropriate candidates for the degradation of chitinous crustacean waste from the seafood industry as they dominated in the biofilms developed on the shrimp's exoskeleton in natural sea water with no added substrates and the degradation of the shrimp exoskeleton was also evidenced.

Dynamics of biofilm formation by Listeria monocytogenes on stainless steel under mono-species and mixed-culture simulated fish processing conditions and chemical disinfection challenges.

The progressive ability of a six-strains L. monocytogenes cocktail to form biofilm on stainless steel (SS), under fish-processing simulated conditions, was investigated, together with the biocide tolerance of the developed sessile communities. To do this, the pathogenic bacteria were left to form biofilms on SS coupons incubated at 15°C, for up to 240h, in periodically renewable model fish juice substrate, prepared by aquatic extraction of sea bream flesh, under both mono-species and mixed-culture conditions. In the latter case, L. monocytogenes cells were left to produce biofilms together with either a five-strains cocktail of four Pseudomonas species (fragi, savastanoi, putida and fluorescens), or whole fish indigenous microflora. The biofilm populations of L. monocytogenes, Pseudomonas spp., Enterobacteriaceae, H2S producing and aerobic plate count (APC) bacteria, both before and after disinfection, were enumerated by selective agar plating, following their removal from surfaces through bead vortexing. Scanning electron microscopy was also applied to monitor biofilm formation dynamics and anti-biofilm biocidal actions. Results revealed the clear dominance of Pseudomonas spp. bacteria in all the mixed-culture sessile communities throughout the whole incubation period, with the in parallel sole presence of L. monocytogenes cells to further increase (ca. 10-fold) their sessile growth. With respect to L. monocytogenes and under mono-species conditions, its maximum biofilm population (ca. 6logCFU/cm2) was reached at 192h of incubation, whereas when solely Pseudomonas spp. cells were also present, its biofilm formation was either slightly hindered or favored, depending on the incubation day. However, when all the fish indigenous microflora was present, biofilm formation by the pathogen was greatly hampered and never exceeded 3logCFU/cm2, while under the same conditions, APC biofilm counts had already surpassed 7logCFU/cm2 by the end of the first 96h of incubation. All here tested disinfection treatments, composed of two common food industry biocides gradually applied for 15 to 30min, were insufficient against L. monocytogenes mono-species biofilm communities, with the resistance of the latter to significantly increase from the 3rd to 7th day of incubation. However, all these treatments resulted in no detectable L. monocytogenes cells upon their application against the mixed-culture sessile communities also containing the fish indigenous microflora, something probably associated with the low attached population level of these pathogenic cells before disinfection (<102CFU/cm2) under such mixed-culture conditions. Taken together, all these results expand our knowledge on both the population dynamics and resistance of L. monocytogenes biofilm cells under conditions resembling those encountered within the seafood industry and should be considered upon designing and applying effective anti-biofilm strategies.

Collagen from the Marine Sponges Axinella cannabina and Suberites carnosus: Isolation and Morphological, Biochemical, and Biophysical Characterization.

In search of alternative and safer sources of collagen for biomedical applications, the marine demosponges Axinella cannabina and Suberites carnosus, collected from the Aegean and the Ionian Seas, respectively, were comparatively studied for their insoluble collagen, intercellular collagen, and spongin-like collagen content. The isolated collagenous materials were morphologically, physicochemically, and biophysically characterized. Using scanning electron microscopy and transmission electron microscopy the fibrous morphology of the isolated collagens was confirmed, whereas the amino acid analysis, in conjunction with infrared spectroscopy studies, verified the characteristic for the collagen amino acid profile and its secondary structure. Furthermore, the isoelectric point and thermal behavior were determined by titration and differential scanning calorimetry, in combination with circular dichroism spectroscopic studies, respectively.

Vertebrae length and ultra-structure measurements of collagen fibrils and mineral content in the vertebrae of lordotic gilthead seabreams (Sparus aurata).

Skeletal deformities of gilthead seabream (Sparus aurata) are a major factor affecting the production cost, the external morphology and survival and growth of the fish. Adult individuals of S. aurata were collected from a commercial fish farm in Greece and were divided into two groups: one with the presence of lordosis, a skeletal deformity, and one without any skeletal deformity. Fishes were X-rayed, and cervical, abdominal and caudal vertebrae lengths were measured. Vertebrae were taken from the site of the vertebral column where lordosis occurred. One part was decalcified and prepared for collagen examination with transmission electron microscopy, and the rest were incinerated, and the Ca and P contents were measured. The stoichiometries of the samples were obtained by EDS (Energy Dispersive Spectroscopy). The same procedure was followed for fish without skeletal deformities (vertebrae were taken from the middle region of the vertebral column). The decalcified vertebrae parts were examined with TEM, collagen micrographs were taken and the fibrils' periods and diameters were measured. There were no significant differences for both Ca and P or the collagen fibrils' periods between the two fish groups. The mean lengths of the cervical, abdominal and caudal vertebrae where lordosis occurred were similar to the lengths of the respective regions of the individuals without the skeletal deformity. The TEM examination showed a significantly smaller mean vertebrae collagen fibril diameter from the fishes with lordosis compared with those from the controls, revealing the significance of collagen to bone structure.

Collagen fibrils in cultured and wild sea bream (Sparus aurata) Liver. An electron microscopy and image analysis study.

This study aims to measure liver collagen fibril diameter in cultured and wild sea breams (Sparus aurata). Cultured sea breams were fed three isonitrogenous diets. The organically produced feed contained sustainable certified fish meal (45%), fish oil (14%), and organic certified wheat; the laboratory feed contained fish meal (45%), fish oil (14%), wheat meal, and soya meal; and the commercial feed included fish meal (46%), fish oil (17%), soya meal, wheat meal, and corn gluten meal. The organic diet had higher amounts of vitamins A, C, and E; specific amino acids; and minerals that enhanced the biosynthesis of collagen. This study shows that fish fed the organic feed had significantly bigger collagen fibril diameters than the fish fed the conventional feed. Furthermore, the organically fed fish had similarly sized collagen fibril diameters as wild fish. More research is needed to understand the long-term effects and the mechanism and function of fish collagen peptide intake on lipid absorption and metabolism; and to identify dietary regimes that are able to improve whole body lipid profiles and suppress the transient increase of plasma triglycerides.

Collagen fibril diameter in relation to bone site and to calcium/phosphorus ratio.

The collagen fibril diameter was measured in cortical bone samples from the femoral neck, rear and front tibia of female and male rats and rabbits using electron microscopy analysis. In most cases, statistically significant differences in mean fibril diameter values between different bone sites were detected. The order of magnitude for the above structural parameter was the same for both genders in both experimental species. In rats, the greatest mean diameter value was that for the femoral, while in rabbits, the one for the rear tibia demonstrating a dependence on bone use and life style. An important aspect was the agreement between these observations and the mean values for Ca/P ratio, as observed in previous experiments, in the same bone sites and animals. Collagen fibril diameter and Ca/P ratio can both serve as indexes of bone quality.

Collagen fibril diameter in relation to bone site. a quantitative ultrastructural study.

The collagen fibril diameter was measured in cortical bone samples from the femoral neck, rear and front tibia of rats using electron microscopy analysis. Statistically significant differences (0.001

Bone calcium, phosphorus detection by Auger electron spectroscopy.

Auger electron spectroscopy was used to detect calcium and phosphorus of cortical bone from rat femoral neck and rear tibia. Spectra were taken from bone pieces as well as from disks prepared from grinded bone material. Experimental conditions were found whereby the samples could be analyzed without conductive coatings. The results of this preliminary investigation demonstrate that Auger electron spectroscopy can be used to study bone mineral elements.