Acclimation to higher temperature and antioxidant supplemented diets improved rainbow trout (Oncorhynchus mykiss) resilience to heatwaves.

Coldwater species are challenged with increasing water temperatures and fluctuations over their upper thermal limits. This study evaluated the potential of acclimation to higher temperature and dietary antioxidants capacity to mitigate the adverse effects of heat shocks in rainbow trout. To this end, rainbow trout fingerlings were acclimated at optimal (14 °C) and high (20 °C) temperatures and fed on selenium (5 mg/kg) and polyphenol (2 g/kg) supplemented diets for 60 days and then were exposed to heat shocks by increasing water temperature up to 30 °C. Growth performance, survival rate, haemato-immunological parameters, and expression of HSP70α, HSP70ß, HSP90ß, and IL-1ß genes were measured to evaluate the hypothesises. The rainbow trout acclimated to 20 °C and fed on antioxidants supplemented diets showed a significantly higher aftershock survival rate. Moreover, fish acclimated to higher temperature showed higher red blood cell counts as well as serum total protein and albumin during the acclimation trial and heat shocks phase. Acclimation to higher temperature and feeding on antioxidants remarkably enhanced fish immune and antioxidant capacity in comparison to fish adapted to cold water and fed on the basal diet measured by improved respiratory burst and lysozyme activities and upregulation of IL-1ß expression during exposure of fish to heat shocks. Furthermore, fish acclimated to higher temperature, especially those fed on antioxidant supplemented diets, showed lower expression levels of HSPs genes during the heat shock phase, indicating that high heat shocks were less stressful for these fish in comparison to cold water acclimated fish. This finding was also supported by lower cortisol levels during heat shocks in fish acclimated to higher temperature. In conclusion, the results of this study indicated that acclimation to higher temperature and/or fed on diets supplemented by selenium and polyphenol, can help to mitigate the adverse effects of the heat shock in rainbow trout.

Genetic species identification and population structure of grouper Epinephelus coioides (Hamilton, 1822) collected from fish markets along the Persian Gulf and the Oman Sea.

Many ecologically important and valuable fisheries marine species have been misidentified in terms of both the statistical data and market demand. Correct identification at the species level and the population genetic structure of the orange-spotted grouper (Epinephelus coioides), a precious fish in the Persian Gulf and the Oman Sea, was tested using mitochondrial cytochrome oxidase subunit I (DNA barcoding) and D-loop sequencing. The results revealed that the Epinephelus species found in the region, including E. coioides, E. bleekeri, E. polylepis, and E. chlorostigma were all mistakenly grouped together and identified as only E. coioides. Moreover, the analysis of molecular variance (AMOVA) of E. coioides samples using the D-loop showed a significantly unique genetic structure (ΦST = 0.068, p < 0.001) within the E. coioides population throughout the Persian Gulf and the Oman Sea, with the pairwise genetic difference between sampling locations in UAE and the Iranian coast. Moreover, D-loop sequences analysis showed two distinct haplotype groups scattered among the sampling locations, which did not correlate with the geographic distance between the sampling locations. These findings indicate that the issue of misidentification should be highlighted in the management and conservation of E. coioides. As this type of misidentification is likely to happen to other threatened marine species as well, the efficacy of using genetic markers for the correct identification, both at the species and the population level, is vital.

Removal of nitrate and phosphate from simulated agricultural runoff water by Chlorella vulgaris.

Microalgae such Chlorella vulgaris can effectively absorb nitrate and phosphate from contaminated water. This work characterized nitrate and phosphate removal from simulated agricultural runoff using C. vulgaris. Statistically designed experiments were used to model the following responses: (1) algal growth; (2) nitrate removal; (3) phosphate removal; (4) protein in the algal biomass; (5) chlorophyll content of the biomass; (6) the biomass phenolics content; and (7) the free radical scavenging antioxidant activity of the biomass. These response were modelled for the following key experimental factors: initial nitrate concentration in the simulated runoff (1080-3240 mg L-1, as NaNO3), initial phosphate concentration (20-60 mg L-1, as K2HPO4), photoperiod (8-24 h of light/day) and culture duration (5-15 days). The validated models were used to identify the factor levels to maximize the various responses. Nitrate removal was maximized at 85.6% when initial nitrate and phosphate concentrations were 2322 mg L-1 and 38 mg L-1 (N:P atom ratio ≈ 125:1), respectively, with a 17.2 h daily photoperiod in a 13-day culture. Phosphate removal was maximized at 95% when the initial nitrate and phosphate concentrations were 1402 mg L-1 and 56.7 mg L-1 (N:P ≈ 51:1), respectively, with a 15.7 h daily photoperiod in a 14.7-day culture. At least ~14 h of a daily photoperiod and a ~11-day culture period were required to maximize all the studied responses. C. vulgaris is edible and may be used as animal feed. Nutritional aspects of the biomass were characterized. Biomass with more than 24% protein could be produced. Under the best conditions, the chlorophyll (potential food colorants) content of the biomass was 8.5% and the maximum level of total phenolics (antioxidants) in the biomass was nearly 13 mg gallic acid equivalent g-1.

Occurrence and characterization of microplastics in white shrimp, Metapenaeus affinis, living in a habitat highly affected by anthropogenic pressures, northwest Persian Gulf.

Microplastics (MPs) are a modern societal concern and recognized as a growing environmental hazard by careless disposal. This study aimed to assess the MPs content in white shrimp (Metapenaeus affinis) inhabiting in a natural habitat affected by high anthropogenic pressures, and recognize if the shrimp could be a suitable bioindicator for MPs pollution. To assess spatial changes of MPs presence in shrimps, sampling was carried out by a trawl net from 13 stations across the entire Musa Bay. Tissues of shrimps were examined for MPs following floatation, digestion, microscopic observation and identified by Confocal Raman Spectroscopy. MPs were observed in the shrimps of all stations. The average MPs abundance was 1.02 items/g of digestive tissues. About 37% of recorded MPs in M. affinis samples exceeding 500 µm that could be related to surface area and stickiness as notable controls beyond ingestion. The dominant shape of MPs was fiber, followed by film. Five different colors were recorded in tissues of M. affinis samples, and the white/transparent MPs were the most abundant, followed by blue and black. In addition, a wide range of recorded colors of MPs in the study area could suggest a variety of sources of MPs. Confocal Raman Spectroscopy confirmed that polyethylene terephthalate (46%), polypropylene (27%) and polystyrene (27%) were dominant polymers. As the average annually consumption of shrimp in the region is 2.3 g/person/day, therefore each person could consume 857 MPs per year. In conclusion, the results of this research provide a detailed and useful information for a better understanding of MPs contamination in the region and suggest Jinga shrimp as a suitable species for monitoring MPs in marine ecosystems.

Immunomodulation, antioxidant enhancement and immune genes up-regulation in rainbow trout (Oncorhynchus mykiss) fed on seaweeds included diets.

This study investigated the stimulatory effects of dietary inclusion of Gracilariopsis persica (GP), Hypnea flagelliformis (HF) and Sargassum boveanum (SB) on immune indices, antioxidant capability and immune related genes expression of rainbow trout (Oncorhynchus mykiss). Seven iso-nitrogenous and iso-caloric diets with 0, 5 and 10% of each macroalgae were prepared and fed to rainbow trout juveniles for 83 days. Serum lysozyme (Lyz) and respiratory burst activity (NBT) along with activity of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) and expression of LyzII, TNFα and IL-1ß genes in head kidney samples were determined by days 47 and 83. Our results revealed that dietary inclusion of seaweeds improved fish immune status. Long term feeding of fish on seaweed contained diets (except for GP10) improved serum Lyz activity in comparison to control group. Similarly, extended feeding on GP5 and HF10 and HF10 included diets improved SOD and POD levels, respectively. Genes expression studies revealed that seaweeds contained diets noticeably enhanced expression of LyzII, TNFα and IL-1ß in comparison to control fish. However, results revealed that such stimulatory effects were more evident at lower dietary inclusion level and shorter feeding time. In conclusion, the results depicted that dietary inclusion of the seaweeds effectively improved serum immune indices and head kidney antioxidant status and immune related genes expression in a time and dose dependent manner.

Implications of anthropogenic effects on the coastal environment of Northern Persian Gulf, using jinga shrimp (Metapenaeus affinis) as indicator.

The distribution and sources of PAHs in jinga shrimp (Metapenaeus affinis), and human health risks due to consumption was evaluated in collected samples from the Musa Bay, Persian Gulf. The total concentration of PAHs (∑16PAHs) ranged from 10 to 144 µg kg-1 dry weight, indicating low to moderate level of pollution. The PAHs were dominated by three- (41%) and two-ring (38%) compounds. Source identification analyses indicated the PAH pollution mostly originated from petroleum inputs. A preliminary evaluation of human health risk using chronic daily intake, hazard index, benzo[a]pyrene-equivalent (BaPeq) concentration (PEC) as well as the incremental life cancer risk and non-cancer risk assessment suggest low potential health risk for consumers of the Metapenaeus affinis. However, the results indicate minimal risks associated with the intake of PAHs via shrimp consumption, but long-term monitoring is required to evaluate the changes in ecological and human health impacts of contaminants in the region. MAIN FINDING: PAHs in Metapenaeus affinis from Musa bay, which influenced by anthropogenic activities were low to moderate level of pollution. Human health risk indicates low potential health risk for consumers.

Long-term effects of three probiotics, singular or combined, on serum innate immune parameters and expressions of cytokine genes in rainbow trout during grow-out.

The long-term effects of three dietary probiotics on rainbow trout during grow-out (mean body weight = 250 ±â€¯50 g) were investigated by feeding for 130 days on eight diet treatments supplemented with Lactobacillus buchneri, L. fermentum and Saccharomyces cerevisiae at 107 CFU/g, singularly or in combination. Fish samples were taken for biochemical and immunological analysis in addition to growth performance indices at days 30 and 130 of the experiment. The expression levels of TNF-α and IL-1ß genes were also measured at day 130. A positive effect on food conversion was observed in rainbow trout with dietary inclusion of S. cerevisiae (P < 0.05) over 130 days. Also, the total number of white blood cells and their differential count (blood neutrophils, lymphocytes and monocytes), as well as respiratory burst activity were all significantly affected by different treatments at 130 days (P < 0.05). Moreover, at 130 days there was a significant increase in the expression of TNF-α and IL-1ß in yeast present treatment compared to the control group (P < 0.05), but no significant difference in the combined probiotic treatments from control group. Yeast and L. buchneri showed a contrary effect on the immune gene expression regulation. Serum cholesterol was significantly lower in all treatments receiving yeast as a dietary probiotic, either alone or in combination with other probiotics. However, none of the probiotic treatments had a significant effect on trout growth performance, or total protein, albumin, globulin, triglyceride and the red blood cell count after 30 or 130 days. Overall, the results suggest that inclusion of a single dietary probiotic, especially S. cerevisiae, in rainbow trout during grow-out has a greater positive effect than combinations of probiotics on the immune system.

Antibacterial activity of Oliveria decumbens against Streptococcus iniae in Nile tilapia (Oreochromis niloticus) and its effects on serum and mucosal immunity and antioxidant status.

The aims of this study were to investigate the antibacterial, immunostimulatory and antioxidant properties of different derivatives of Oliveria decumbens, in vitro and in vivo. The GC-MS spectrometry analysis showed γ-terpinene as the most frequent compound in essential oil, whereas carvacrol and thymol were the most common ones in aromatic water. Plant essential oil and hydroethanolic extract showed a positive in vitro bactericidal activity against Streptococcus iniae as evaluated by disc diffusion, minimum inhibitory concentration and minimum bactericidal concentration methods. Also, in vivo resistance against S. iniae and immune and antioxidant responses of Nile tilapia (Oreochromis niloticus) were assayed after 60 days treatment with O. decumbens derivatives. Plant hydroethanolic extract and essential oil and their 1:1 combination were added to diet at 0 (negative control), 0.01, 0.1 and 1% (w:w). The plant aromatic water at doses of 0.0312, 0.0625 and 0.1250% were also used as bath treatment. The results showed that aromatic water at lowest dose was more effective than other treatments in increment of fish resistance against S. iniae (7.14% mortality in comparison with 50% mortalities in control fish) and modulation of post-challenge respiratory burst activity. The bactericidal activity and biochemical contents of skin mucus did not change significantly among treatments. The levels of superoxide dismutase and catalase antioxidant enzymes activities in spleen tissue were significantly higher in treatments received extract, essential oils and their combination in comparison to other groups, while treatments did not affect peroxidase level. In conclusion, administration of different derivatives of Oliveria decumbens showed remarkable antibacterial activity against streptococcosis and enhanced antioxidant status and post-challenge immunity in Nile tilapia.

Changes in growth, blood immune parameters and expression of immune related genes in rainbow trout (Oncorhynchus mykiss) in response to diet supplemented with Ducrosia anethifolia essential oil.

An 8- week feeding trial was conducted to investigate the effects of Ducrosia anethifolia essential oil on growth, blood immune parameters and immune related genes expression in juvenile rainbow trout (Oncorhynchus mykiss). Fish were allocated into 4 groups and fed on diet containing different levels of essential oil (0, 0.001, 0.01 and 0.1%) to apparent satiation in 30 min 3 times daily. Growth and immunological parameters were measured every ten days and tissue samples were taken from kidney and spleen on days 10, 30 and 50 to study the expression of IL-1ß and TNF-α. The changes of measured parameters in different treatments and over sampling time series were statistically analysis based on repeated measurement method (P < 0.05). Results showed that growth did not affected by essential oil at different treatments. The highest level of hematocrit was observed in 0.001 treatment. The mean of RBC showed no significant differences among treatments. The mean of WBC in 0.01 and 0.1 groups were higher than those in 0.001 and control groups. Total protein, albumin and globulin and serum bactericidal activity showed no significant difference in different treatments. Neither treatments nor sampling times affected serum lysozyme activity. The highest mean of respiratory burst activity was observed in 0.01 group. The highest expression of both IL-1ß and TNF-α genes in kidney was observed at 0.001 dose on day 30, while in spleen, the highest expression of IL-1ß and TNF-α was obtained on day 30 at doses 0.1 and 0.01%, respectively. In conclusion the results of this study showed that feeding with lower and medium level of D. anethifolia for 30 days led to immunostimulatory effects in juvenile rainbow trout.

Differential expression of subunits of 20ß-hydroxysteroid dehydrogenase during gametogenesis in rainbow trout (Oncorhychus mykiss).

The patterns of expression of two subunits of 20ß-hydroxysteroid dehydrogenase (20ß-HSD), key enzyme involved in the biosynthesis and activation of steroid hormones, were examined in rainbow trout by using a combination of quantitative real-time PCR and in-situ hybridization. The expression of targeted genes was examined in mRNA extracted from different tissues at different gonadal stages in male and female trout. Both subunits of 20ß-HSD were found to be widely distributed in tissues. The highest expression of 20ß-HSD A was found in intestine followed by skin, stomach, liver and gills, whereas, the highest expression of 20ß-HSD B was observed in stomach followed by head kidney, ovary - at late vitellogenesis stage- and trunk kidney. In ovarian tissue 20ß-HSD A was highly expressed in mature oocytes, and the highest expression of 20ß-HSD B was in ovary at late vitellogenesis stage. There were no differences in the level of expression of either subunit among groups of rainbow trout at different stages of maturational competence. In male fish, 20ß-HSD A was highly expressed in testis stage I in contrast to 20ß-HSD B which was highly expressed in testis stage VIII. In situ- hybridization results showed that the 20ß-HSD gene was highly expressed in gastrointestinal organs, while only slightly expressed in the gonadal tissue of fish at stage 62day-post-fertilization (dpf). Overall, the results confirm the ubiquitous presence of 20ß-HSD among tissues in rainbow trout with relatively minor fluctuations in expression associated with reproductive cycles which collectively suggests a wider metabolic role of these enzymes than just an association with the synthesis of control hormones for reproduction.

Comparison of methods to improve induction of spermiation in wild-caught carp (Cyprinus carpio carpio), a threatened species from the Caspian Sea basin.

Wild carp (Cyprinus carpio carpio) forms the basis of an important fishery in the Southern Caspian Sea Basin which is increasingly underpinned by the release of cultured juveniles. A significant bottleneck to hatchery-rearing of juveniles is the spermiation of male broodstock. Therefore, four approaches to improving spermiation were investigated. The effectiveness of two delivery methods for the sustained release of salmon gonadotropin releasing hormone analogue (sGnRHa; i.e., via intramuscular cholesterol pellet vs emulsion injection) on the spermiation success and duration, sperm quality and quantity over 14days in wild-caught carp were compared to single injection of sGnRHa with Pimozide(®) (Linpe method) or carp pituitary extract (CPE). The consequence of the spermiation treatments on resulting embryonic quality was evaluated for subsequent fertilization and hatching success from wild male carp (mean weight±S.D. 1021±112g). All hormonal treatments, except for Linpe method, led to 100% spermiation of treated fish compared to only 25% in the control with no hormone intervention. The duration of spermiation, as well as the various quantitative variables of the sperm and the mean total sperm production were all generally improved with the sustained hormone delivery compared with the acute treatments. The GnRHa-FIA was the most effective method for improving spermiation.

Concentration of some heavy metals in rice types available in Shiraz market and human health risk assessment.

This investigation was conducted to survey the levels of some heavy metals such as cadmium, lead, chromium, nickel and cobalt in domestic cultivated and imported rice sold on the Shiraz - Iran markets. The potential human health risk assessment was conducted by considering estimated weekly intake (EWI) of toxic metals from eating rice and compared calculated values with provisional tolerable weekly intake (PTWI). The mean values for lead and cadmium in domestic cultivated and imported rice were considerably higher than allowable limits set by FAO/WHO. In combination of recent rice consumption data, the estimated weekly intakes of toxic element were calculated for Iranian population. EWI for cadmium, nickel, chromium through imported and domestic cultivated rice consumption was lower than the PTWI. The EWI for lead were considerably higher than other measured toxic metals. The highest mean level of EWI for lead was observed in some imported rice samples (25.76 µg/kg body weight).