Integrative approach of histopathology and histomorphometry of common carp (Cyprinus carpio L.) organs as a marker of general fish health state in pond culture

The semi‐intensive system of common carp monoculture in earthen ponds is currently the most practiced fish rearing system in several countries in Eastern Europe. This system is based on the addition of supplemented feed to ponds to achieve faster growth. In this study, the general state of the fish health was monitored when three different types of feeds were used: grain cereals (CF), commercial pelleted feed and extruded feed. The markers used were: (i) the histopathological index (HP) of the main organs: gills, liver and kidney; and (ii) the histomorphometric parameters of the hepatocytes and enterocytes to assess the state of the digestive system in fish fed different types of supplemented feed. At the end of the experiment, the fish were in good health, since irreversible lesions were not noticed to a greater extent in any of the fish organs. There were also no other severe lesions present. However, the results showed differences in the HP indices of the liver and gills, as well as the total HP index, indicating a better health status of the fish in the CF group. The histomorphometric results showed larger cytoplasmic areas of hepatocytes in fish fed CF, which is attributed to glycogen accumulation and higher numbers of goblet cells in the distal intestines of fish fed compound feeds. This can be attributed to slight inflammation.     

The influence of sex on biochemical markers of bone turnover in dogs

Biochemical markers of bone turnover have been shown to be useful as inexpensive and noninvasive tools for monitoring skeletal health. The reference range for bone markers in dogs has been set by different age groups. However, other sources of biological variations were not fully investigated in dogs. To explore whether sex influences the interpretation of bone marker data we examined serum bone markers in 33 male and 25 female dogs. The bone markers selected for this study were: bone alkaline phosphatase (BALP) and osteocalcin (OC) as indicators of bone formation, and C-terminal telopeptide (CTx) of type I collagen as marker of bone resorption. All concentrations of bone markers were lower, but still within the reference range reported for dogs. We found statistically significant differences of the median OC and CTx serum concentrations between males and females. The results of this study suggest that there are sex differences in biochemical markers of bone turnover in dogs which should be considered in interpretation of bone marker data.     

Interpreting the dependence of soil respiration on soil temperature and water content in a boreal aspen stand

Continuous half-hourly measurements of soil CO₂ efflux made between January and December 2001 in a mature trembling aspen stand located at the southern edge of the boreal forest in Canada were used to investigate the seasonal and diurnal dependence of soil respiration (R_s) on soil temperature (T_s) and water content (θ). Daily mean R_s varied from a minimum of 0.1 μmol m⁻² s⁻¹ in February to a maximum of 9.2 μmol m⁻² s⁻¹ in mid-July. Daily mean T_s at the 2-cm depth was the primary variable accounting for the temporal variation of R_s and no differences between Arrhenius and Q₁₀ response functions were found to describe the seasonal relationship. R_s at 10 °C (R_s10) and the temperature sensitivity of R_s (Q_10Rs) calculated at the seasonal time scale were 3.8 μmol m⁻² s⁻¹ and 3.8, respectively. Temperature normalization of daily mean R_s (R_sN) revealed that θ in the 0–15 cm soil layer was the secondary variable accounting for the temporal variation of R_s during the growing season. Daily R_sN showed two distinctive phases with respect to soil water field capacity in the 0–15 cm layer (θ_fc, 0.30 m³ m⁻³): (1) R_sN was strongly reduced when θ decreased below θ_fc, which reflected a reduction in microbial decomposition, and (2) R_sN slightly decreased when θ increased above θ_fc, which reflected a restriction of CO₂ or O₂ transport in the soil profile.Diurnal variations of half-hourly R_s were usually out of phase with T_s at the 2-cm depth, which resulted in strong diurnal hysteresis between the two variables. Daily nighttime R_s10 and Q_10Rs parameters calculated from half-hourly nighttime measurements of R_s and T_s at the 2-cm depth (when there was steady cooling of the soil) varied greatly during the growing season and ranged from 6.8 to 1.6 μmol m⁻² s⁻¹ and 5.5 to 1.3, respectively. On average, daily nighttime R_s10 (4.5 μmol m⁻² s⁻¹) and Q_10Rs (2.8) were higher and lower, respectively, than the values obtained from the seasonal relationship. Seasonal variations of these daily parameters were highly correlated with variations of θ in the 0–15 cm soil layer, with a tendency of low R_s10 and Q_10Rs values at low θ. Overall, the use of seasonal R_s10 and Q_10Rs parameters led to an overestimation of daily ranges of half-hourly R_s (ΔR_s) during drought conditions, which supported findings that the short-term temperature sensitivity of R_s was lower during periods of low θ. The use of daily nighttime R_s10 and Q_10Rs parameters greatly helped at simulating ΔR_s during these periods but did not improve the estimation of half-hourly R_s throughout the year as it could not account for the diurnal hysteresis effect.     

Optimisation of sodium and potassium concentrations and pH in the artificial seminal plasma of common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.

The effect of sodium and potassium concentrations as well as optimal pH on the motility of common carp Cyprinus carpio L. sperm during short-term storage in artificial seminal plasma (ASP) was investigated. Sperm was collected from individual males (n?=?5) and each sample diluted tenfold (1:9) in ASP (sperm:extender) containing 2 mM CaCl₂, 1 mM Mg₂SO₄ and 20 mM Tris at pH 8.0 and supplemented by the following concentrations of sodium and potassium (mM/mM): 0/150, 20/130, 40/110, 75/75, 110/40, 130/20 and 150/0. The osmolality of all ASP variants was set at 310 mOsm kg^?1. Sperm motility was measured using a CASA system during 72 h of storage. Immediately after dilution, sperm motility was high (90%) both in each variant and in the control group (fresh sperm). After 72-h storage, the highest sperm motility was noted in ASP containing 110 mM NaCl and 40 mM KCl. No differences were found in the motility of samples preserved within the pH range of 7.0–9.0. Our data suggest that for the short-term storage of common carp sperm, whereas the pH of the solution does not play a crucial role, a specific potassium concentration of around 40 mM is required.     

Effect of supplemental feed type on water quality,plankton and benthos availability and carp (Cyprinus carpio L.) growth in semi‐intensive monoculture ponds

A four‐and‐a‐half months study was conducted in nine earthen ponds to evaluate the addition of different supplemental feeds as a management tool for enhancing natural food availability and common carp growth, while maintaining optimal water quality in the semi‐intensive system. Three supplemental feeds were used: commercial extruded and pelleted feed with 25% protein and 7% fat and cereals. The type of supplemental feed did not influence water quality, except hardness, but significantly affected abundance of cyanobacteria, natural food availability and common carp growth. The use of pelleted feed was related to the lowest abundance of cyanobacteria in the ponds. For the two groups of large zooplankton, Cladocera and Copepoda, abundances were higher in the ponds with pelleted feed compared with the ponds where cereals and extruded feed were used. The abundance of benthic macroinvertebrates in the treatment with pelleted feed was three times higher than in the other two treatments. The results of this study indicate that pelleted feed can help farmers not only as a source of nutrients for carp growth but also indirectly as a management tool for maintaining ecological stability and control of cyanobacterial bloom in ponds.     

Preservation of common carp germ cells under hypothermic conditions: Whole tissue vs isolated cells

The aim of this study was to optimize the conditions for hypothermic storage of spermatogonial stem cells (SSCs) and oogonial stem cells (OSCs) of common carp Cyprinus carpio. This was conducted by storing gonadal tissue or isolated cells for 24 hr under hypothermic conditions in the first experiment and by testing two different storage media (L‐15 or DMEM supplemented with 10% FBS and 25 mM HEPES) and regular medium change (every 4 days) during two weeks of hypothermic storage in the second experiment. During the first 24 hr, isolated cells showed no decrease in viability, while cells obtained from hypothermically stored tissues displayed significantly lower viability after only 6 hr (Tukey's HSD, p < 0.01) indicating that hypothermic storage of isolated cells is superior to storing tissue pieces. The 2‐week trial demonstrated that storage media have a profound influence, while regular medium exchange does not have a positive effect on cell viability. Viability of SSCs and OSCs after two weeks was approximately 40% and 25%, respectively; however, survival of ~70% was obtained after 10 days of storage for SSCs and 7 days for OSCs. Hypothermic storage developed in this study has many practical applications during the development of surrogate broodstock technologies for common carp, but also in carp hatcheries and for the conservation of genetic resources of closely related cyprinid species.     

Sulfated Polysaccharide Extracted from the Green Algae Codium bernabei: Physicochemical Characterization and Antioxidant,Anticoagulant and Antitumor Activity

Codium bernabei is a green alga that grows on Chilean coasts. The composition of its structural polysaccharides is still unknown. Hence, the aim of this work is to isolate and characterize the hot water extracted polysaccharide fractions. For this purpose, the water extracts were further precipitated in alcohol (TPs) and acid media (APs), respectively. Both fractions were characterized using different physicochemical techniques such as GC-MS, GPC, FTIR, TGA, and SEM. It is confirmed that the extracted fractions are mainly made of sulfated galactan unit, with a degree of sulfation of 19.3% (TPs) and 17.4% (ATs) and a protein content of 3.5% in APs and 15.6% in TPs. Other neutral sugars such as xylose, glucose, galactose, fucose, mannose, and arabinose were found in a molar ratio (0.05:0.6:1.0:0.02:0.14:0.11) for TPs and (0.05:0.31:1.0:0.03:0.1:0.13) for ATs. The molecular weight of the polysaccharide samples was lower than 20 kDa. Both polysaccharides were thermally stable (Tonset > 190 °C) and showed antioxidant activity according to the ABTS^•+ and DPPH tests, where TPs fractions had higher scavenging activity (35%) compared to the APs fractions. The PT and APTTS assays were used to measure the anticoagulant activity of the polysaccharide fractions. In general, the PT activity of the TPs and APs was not different from normal plasma values. The exception was the TPs treatment at 1000 µg mL⁻¹ concentration. The APTTS test revealed that clotting time for both polysaccharides was prolonged regarding normal values at 1000 µg mL⁻¹. Finally, the antitumor test in colorectal carcinoma (HTC-116) cell line, breast cancer (MCF-7) and human leukemia (HL-60) cell lines showed the cytotoxic effect of TPs and APs. Those results suggest the potential biotechnological application of sulfate galactan polysaccharides isolated from a Chilean marine resource.     

Interspecific germ cell transplantation: a new light in the conservation of valuable Balkan trout genetic resources?

Interspecific transplantation of germ cells from the brown trout Salmo trutta m. fario and the European grayling Thymallus thymallus into rainbow trout Oncorhynchus mykiss recipients was carried out in order to improve current practices in conservation of genetic resources of endangered salmonid species in the Balkan Peninsula. Current conservation methods mainly include in situ efforts such as the maintenance of purebred individuals in isolated streams and restocking with purebred fingerlings; however, additional ex situ strategies such as surrogate production are needed. Steps required for transplantation such as isolation of high number of viable germ cells and fluorescent labeling of germ cells which are to be transplanted have been optimized. Isolated and labeled brown trout and grayling germ cells were intraperitoneally transplanted into 3 to 5 days post hatch rainbow trout larvae. Survival of the injected larvae was comparable to the controls. Sixty days after transplantation, fluorescently labeled donor cells were detected within the recipient gonads indicating successful incorporation of germ cells (brown trout spermatogonia and oogonia—27%; grayling spermatogonia—28%; grayling oogonia—23%). PCR amplification of donor mtDNA CR fragments within the recipient gonads additionally corroborated the success of incorporation. Overall, the transplantation method demonstrated in this study presents the first step and a possible onset of the application of the germ cell transplantation technology in conservation and revitalization of genetic resources of endangered and endemic species or populations of salmonid fish and thus give rise to new or improved management strategies for such species.     

Beneficial Effect of Virgin Coconut Oil on Alloxan-Induced Diabetes and Microbiota Composition in Rats

The gut microbiota is composed of a diverse population of obligate and facultative anaerobic microorganisms which are shown to influence host metabolism and immune homeostasis. This study investigated the effects of virgin coconut oil on the weekly fasting glycaemia, daily food and water intake and weekly body mass gain over 16 weeks, as well as the changes in composition of gut microbiota in both non-diabetic and alloxan-induced diabetic rats. Although the intake of virgin coconut oil did not decrease the diabetes-induced hyperglycemia, it affected the secondary parameters, such as food and water intake and average body mass gain. Furthermore, its potential to positively affect the fecal microbiome was proved, since it significantly increased the abundance of probiotic bacteria, such as Lactobacillus, Allobaculum and Bifidobacterium species.