Development and evaluation of a fish‐based index to assess biological integrity of Mediterranean streams

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Decapsulated Artemia Cysts: A Suitable Dietary Supplement for Juvenile Tench (Tinca tinca L.)

There are no specifically formulated dry foods for tench (Tinca tinca L.), which forces farmers to use diets formulated for other fish species. This has major drawbacks, such as high mortality, slow growth, and body deformities. A 120 day experiment was performed with five-month-old juvenile tench (initial mean weight: 0.388 g; total length: 31.78 mm) to evaluate decapsulated Artemia cysts as a supplement to a dry diet for other fish species. Three treatments, differing in the daily supplement, were tested: 1,800 freshly hatched nauplii, 1,800 cysts, and 300 cysts per g of tench biomass. Final survival ranged between 95.3% and 97.9%. Juvenile tench that received the supplement of 1,800 decapsulated Artemia cysts had a specific growth rate (1.28), weight (1.83 g), and total length (52.30 mm) significantly higher than those with the same amount of nauplii. The lowest supplement (300 cysts/g of fish biomass) allowed significantly lower growth and higher condition coefficient (1.40) than the rest. Animals with body deformities (1.06%) were only recorded in the groups that received the lowest cyst supplement. Results showed Artemia cysts are a suitable dietary supplement for juvenile tench, being an advantageous alternative to live nauplii.     

Biochemical,Physical, Chemical,and Microbiological Assessment of Blue Shrimp (Litopenaeus stylirostris) Stored in Ice

Postmortem changes in blue shrimp (Litopenaeus stylirostris) muscle were studied on the basis of biochemical, chemical, physical, and microbiological changes during an 18 day storage period at 0°C. Adenosine 5′-triphosphate (ATP) content, breakdown products, K-value, pH, trimethylamine nitrogen (TMA-N), total volatile basic nitrogen (TVB-N), water holding capacity (WHC), color, and texture (shear force) changes were examined. Also, total mesophilic and psychrophilic bacterial counts were measured. K-value increased linearly (r² = 0.98) from an initial value of 1.37 ± 0.59 to 59.42 ± 6.05% at Day 18. Spoilage indicators TVB-N and TMA-N increased from 29.56 ± 1.33 and 0.69 ± 0.25 to 39.04 and 2.04 ± 0.59 mg of N/100 g at Day 18, respectively; meanwhile, the total viable counts of mesophilic and psychrophilic bacteria increased from 3.48 ± 0.44 and 2.61 ± 0.29 log CFU/g to 6.27 ± 0.21 and 7.14 ± 0.39 log CFU/g, respectively, which indicated spoilage at the end of the storage period. The pH, texture, WHC, and color were affected (p < 0.05) during the storage period. Overall, results indicate that blue shrimp muscle quality was maintained for 12 days of storage in ice.     

Effects of formalin on total haemocytes count and histopathological changes in the shrimp Litopenaeus schmitti (Pérez‐Farfante & Kensley 1997)

Formalin is used in shrimp industry to eliminate ectoparasites and as a disinfectant. However, the effect of formalin on shrimp's defense mechanism is unknown. Several biomarkers were used to assess the immune response of juveniles of the shrimp Litopenaeus schmitti exposed to different concentrations of formalin (10, 25 and 50 mg L^?1) during 24 and 48 h. Formalin concentrations of 25 and 50 mg L^?1 produced a significant reduction in the total haemocyte count at both 24 and 48 h. Peroxidase and phenoloxidase activity did not show variations. The total protein concentration in haemolymph of shrimps was not significantly different between experimental groups. Histological analysis showed hyperplasia in the gills of animals exposed at 10 mg L^?1 for 24 h; however, the exposure at 25 and 50 mg L^?1 caused necrosis in gills, hepathopancreas and the antennal gland.     

The pond's shape matters: differential growth,physiological condition and survival of epibenthic Farfantepenaeus aztecus postlarvae

As a function of the water quality provided by square, circular and oval experimental ponds, the growth, survival and oxygen requirements in epibenthic postlarvae of Farfantepenaeus aztecus were analysed in relation to their routine metabolism and apparent heat increment. Temperature, oxygen concentration, pH and salinity were measured daily in two experimental ponds of each shape. The postlarvae oxygen consumption during two 24‐h cycles, their growth, physiological condition and survival and the productivity in the ponds were estimated. Low values of pH, oxygen concentration and phytobenthos productivity, and reduced postlarvae relative growth and survival were observed in the square ponds. We suggest that the latter results from a deficient water circulation related to the effect of the pond's shape on dissolved oxygen levels and, consequently, on growth and survival. The postlarvae routine metabolism, including feeding, varied between 1.91 and 2.25 mg O₂ h^?1 g^?1 wet weight, whereas the minimum oxygen concentration needed in the ponds is approximately 4.25 mg O₂ L^?1. These conditions were achieved in the oval ponds concurrent with higher survival and growth values, in which individuals distributed randomly, for which we suggest that oval‐shaped ponds could be the most adequate for the culture of this and other penaeid species.     

Estimation of actual evapotranspiration for a drip-irrigated Merlot vineyard using a three-source model

A study was performed in order to evaluate the three-source model (Clumped model) for direct estimation of actual evapotranspiration (ET_a) and latent heat flux (LE) over a drip-irrigated Merlot vineyard trained on a vertical shoot positioned system (VSP) under semi-arid conditions. The vineyard, with an average fractional cover of 30%, is located in the Talca Valley, Region del Maule, Chile. The performance of the Clumped model was evaluated using an eddy covariance system during the 2006/2007 and 2007/2008 growing seasons. Results indicate that the Clumped model was able to predict ET_a with a root mean square error (RMSE), mean bias error (MBE), and model efficiency (EF) of 0.33, −0.15 mm day⁻¹ and 74%, respectively. Also, the Clumped model simulated the daytime variation of LE with a RMSE of 36 W m⁻², MBE of −8 W m⁻², and EF of 83%. Major disagreement (underestimated values) between observed and estimated values of ET_a was found for clear days after rainfall or foggy days, but underestimated values were less than 10% of the data analysis. The results obtained in this study indicate that the Clumped model could be used to directly estimate vine water requirements for a drip-irrigated vineyard trained on a VSP. However, application of the Clumped model requires a good characterization of the drip-irrigated vineyard architecture.     

Salmonid fish viruses and cell interactions at early steps of the infective cycle

A flow cytometric virus-binding assay that directly visualizes the binding and entry of infectious pancreatic necrosis virus (IPNV), infectious haematopoietic necrosis virus (IHNV) and virus haemorrhagic septicaemia virus (VHSV) to several cell lines was established. The highest efficiency of binding was shown by the BF-2 cell line and this was used to study, at the attachment level, the interactions of these cells with salmonid fish viruses in coinfections, and to further determine if the earliest stage of the viral growth cycle could explain the previously described loss of infectivity of IHNV when IPNV is present. Our results demonstrated that IPNV binds to around 88% of cells either in single or dual infections, whereas IHNV attachment always decreased in the presence of any of the other viruses. VHSV binding was not affected by IPNV, but coinfection with IHNV reduced the percentage of virus-binding cells, which suggests competition for viral receptors or co-receptors. Internalization of the adsorbed IHNV was not decreased by coinfection with IPNV, so the hypothetical competence could be restricted to the binding step. Treatment of the cells with antiviral agents, such as amantadine or chloroquine, did not affect the binding of IPNV and VHSV, but reduced IHNV binding by more than 30%. Tributylamine affected viral binding of the three viruses to different degrees and inhibited IPNV or IHNV entry in a large percentage of cells treated for 30 min. Tributylamine also inhibited IHNV cytopathic effects in a dose-dependent manner, decreasing the virus yield by 4 log of the 50% endpoint titre, at 10 mm concentration. IPNV was also inhibited, but at a lower level. The results of this study support the hypothesis that IHNV, in contrast to VHSV or IPNV, is less efficient at completing its growth cycle in cells with a simultaneous infection with IPNV. It can be affected at several stages of viral infection and is more sensitive to the action of antiviral compounds.     

Physiological responses of Quercus oleoides (Schltdl & Cham) to soils contaminated by diesel

ABSTRACT

Oil pollution is a worldwide threat to the environment that affects the development of plants. The effect of soil contaminated by diesel on the physiological responses of seedlings of Quercus oleoides was investigated in two independent experiments. We proposed that physiological performance will decrease when seedlings are exposed to higher concentration of contamination. At the first experiment, levels of pollution with diesel were of 0%, 5%, 10% and 15%, and 0%, 2% and 3.5% in the second one. In the first experiment, photosynthetic rate, stomatal conductance, transpiration and total chlorophyll of Q. oleoides were higher in the control seedling, and lower in treatments of pollution with 5%, 10% and 15% of diesel during 3 days of treatment. Only, seedlings in soil polluted with 5% of diesel survived up to 16 days; they showed a photosynthetic rate of 5 μmol m^?2 s^?1, which was lower than control seedlings (9 μmol m^?2 s^?1). This pattern was observed in stomatal conductance, transpiration and relative water content. Surprisingly, in the second experiment, seedlings showed a higher photosynthetic rate and growth at 2% of diesel-contaminated soil than control seedlings, a phenomenon known as hormesis. In both experiments, soil respiration was proportional to soils contaminated. We concluded that Q. oleoides is highly vulnerable in soils contaminated with above 5% of diesel, but it maintains its physiological activities in soils contaminated below 2%, suggesting that seedlings can grow under low concentration of diesel contaminant, and may be used in phytoremediation of soils with low concentrations of diesel contamination.     

Design and validation of a RT‐qPCR procedure for diagnosis and quantification of most types of infectious pancreatic necrosis virus using a single pair of degenerated primers

Infectious pancreatic necrosis virus (IPNV) is an important virus which affects the salmonid aquaculture industry worldwide; therefore, it is important to develop rapid and reliable methods of diagnosis to detect the disease at early stages. Nowadays, RT‐qPCR is replacing other methods because it provides additional information on the viral load, which is important to have a better understanding of the virus replication level and of the stage of the infection and its risk level. The main problem stems from the high diversity of this virus, which can compromise the reliability of the diagnosis. In this study, we have designed an RT‐qPCR procedure for diagnosis and quantification of IPNV based on a single pair of primers targeted to segment B. The procedure has been validated, in vitro and in vivo, testing two different types of standards against seven reference strains and 23 field isolates from different types. The procedure is reliable for the detection of any type, with a detection limit of 31 TCID₅₀ mL^?1, 50 pfu mL^?1 or 66 RNA copies mL^?1, depending on the standard. All the standard curves showed high reliability (R² > 0.95). The results support the high reliability of this new procedure for the diagnosis and quantification of IPNV.     

Soybean molasses as an organic carbon source in the farming of Litopenaeus vannamei (Boone, 1931) in a biofloc system

Soybean molasses was evaluated as a partial replacement for sugarcane molasses as a carbon source for biofloc development in the superintensive culture of Pacific white shrimp (Litopenaeus vannamei). A 50‐day study was conducted with juvenile (3.2 g) shrimp stocked in 16 800 L tanks at a stocking density of 250 shrimp m^?3. Control of total ammonia concentration was performed by the addition of combined mixtures of soybean and sugarcane molasses to the culture water. Three different molasses treatments were evaluated using different soybean‐to‐sugarcane molasses ratios: 15–85%, 38–62% and 60–40% respectively. The control group was treated only with sugarcane molasses. Water quality, chlorophyll a concentration, heterotrophic bacterial load, Vibrio spp. concentration and zootechnical indexes were all evaluated. Total ammonia concentration was controlled by heterotrophic and chemotrophic pathways. Biofloc formation, as quantified by measuring the total suspended solids, was not altered. The Vibrio spp. concentration showed a significant reduction in treatments with soybean‐to‐sugarcane molasses ratios of 38–62% and 60–40%. All combined mixtures of soybean and sugarcane molasses could maintain water quality and productivity in the superintensive culture of L. vannamei using the biofloc system. Thus, the potential use of a residue from agroindustry as a carbon source in a biofloc culture is demonstrated.