Modelling of Pore Water Nutrient Distribution and Benthic Fluxes in Shallow Coastal Waters (Gulf of Trieste, Northern Adriatic)

Pore waters, extracted monthly from short cores at two sedimentologically and biologically different locations (AA1 and F) in the Gulf of Trieste (Northern Adriatic), were analyzed for NH₄ ⁺, NO₃ ⁺, PO₄ ³⁺, Si(OH)₄, DIC and periodically for DOC, DON and DOP. Nutrient concentrations were used to model vertical profiles using a diffusion-reaction model which included the macrofaunal influence on sediment-water exchange rates. Winter nutrient profiles showed nearly an exponential increase, or decrease in the case of NO₃ ⁺, in nutrient concentrations with depth while the profiles from other seasons exhibited concentration maximum at 3–6 cm, a minimum around 8–10 cm, and then, except for NO₃ ⁺, a gradual increase or constant values. This vertical distribution is attributed to seasonal variations in the benthic infauna activity, mostly composed of polychaetes and bivalves and concentrated in the top 4–5 cm, being less active during the winter. The vertical profiles of DOC and DON showed the vertical distribution described above in all periods, while DOP was similar to that of PO₄ ³. The comparison of modelled fluxes of nutrients across the sediment-water interface at the location AA1 and those measured usingin situ benthic chamber showed quite good agreement for NH₄ ⁺ and PO₄ ³, fluxes but not for NO, and Si(OH)₄. Discrepancies could be caused by a topography effect and for Si(OH)₄ by an additional dissolution of the solid phase balancing the diffusive loss into burrows and lessening the effect of bioturbation.     

Cryopreservation of llama semen using a combination of permeable cryoprotectants

Semen cryopreservation is not available for massive use in South American Camelids (SACs) due to the lack of an efficient protocol and the low pregnancy rates obtained with artificial insemination (AI). The use of a single cryoprotectant (CP) is commonly used in SACs frozen semen. The objective of the study was to evaluate the combined cryoprotective capacity of two permeable CPs at different stages of the cryopreservation protocol in llama semen. Sixteen ejaculates from 4 llama males were analysed, and sperm quality was assayed in raw semen, at 5°C, after equilibration of samples with the CPs and when samples were thawed. The following CPs and combination were used: 6% glycerol (GL), 6% dimethylformamide (DMF) and the combination of both CPs: 3% GL and 3% DMF. A Kruskal–Wallis test and an experimental factorial design, considering one factor with four levels (raw semen, 6% GL, 6% DMF and GL/DMF), were used. Total sperm motility and live sperm with intact acrosomes remained unchanged after equilibration of samples (p > .05). A significant decrease in the percentage of functional membrane, motile and live sperm with intact acrosomes was observed when samples were thawed (GL, DMF and GL/DMF). Nevertheless, the cryopreservation protocols used preserved sperm DNA quality; thus, sperm chromatin condensation and DNA fragmentation were unaffected (p > .05) when GL, DMF and GL/DMF were used. To conclude, no superiority was found between the use of a single or a combination of permeable cryoprotectants to freeze llama semen.     

Sampling strategies for soil water content to estimate evapotranspiration

When the soil water balance method is applied at a field scale, estimation of the spatial variability and confidence interval of actual evapotranspiration is rare, although this method is sensitive to the spatial variability of the soil, and thus to the sampling strategy. This work evaluated the effect of soil sampling strategies for soil water content and water flux at the bottom of the soil profile on the estimation of the daily and cumulative evapotranspirations. To do that, according to the statistical properties of daily measurements in a field experiment with a soybean crop, the water content and flux through the base to the soil profile in space (field scale) and time (daily scale) were simulated. Four different sampling strategies were then compared, and their effects on daily and seasonal cumulative evapotranspirations quantified. Strategy 1 used ten theoretical sites randomly located in the field. The daily water content estimates were assumed to be available each day from these same ten locations, which were located from 0.15 m to 1.55 m in depth, with space steps of 0.10 m. Strategy 2 assumed that daily water content estimates combined two sources: in the 0.00–0.20 m soil layer, ten theoretical sites were selected but changed every day, with thin soil layers for soil moisture sampling, from 1 to 5 cm in thickness. In the 0.20–1.60 m soil layer, the daily water content estimates were assumed to come from the same ten locations (the first soil moisture estimate was located at 0.25 m, and the others were located every 0.10 m until 1.55 m). Strategy 3 used ten theoretical sites located in the field, as in strategy 1, however the water content estimates in the 0.00–0.20-m soil layer were assumed to come from accurate water content measurements (soil layers from 1 to 5 cm in thickness), while for the 0.20–1.60 m soil layer, the strategy was similar to strategies 1 and 2. Strategy 4 used 10 new theoretical locations of measurement every day. Precise water content estimates for thin layers were assumed to be available in the 0.00–0.20 m soil layer as in strategy 2. The layers for water content estimates in the 0.20–1.60 m were similar to those of strategies 1, 2, and 3. Results showed that the spatial variability of the daily actual evapotranspiration may not be negligible, and differences from approximately ±1.0 mm d ^–1 to ±3.0 mm d ^–1 were calculated between the four sampling strategies. Strategy 1 gave the worst results, because variations in the water content of the top soil layers were neglected, and thus the daily evapotranspiration was underestimated. Strategy 2 led to a considerable variability for estimating daily evapotranspiration which was explained by the effect of the spatial variability due to the daily site sampling for the top soil layers (0 to 0.2 m). Strategy 3 appeared to be the best practical compromise between practical field considerations and the necessity to obtain accurate evapotranspiration measurements. The accuracy of daily evapotranspiration could reach ± 0.5 mm d^–1, and could be further improved by increasing the number of measurement sites. The best results were obtained with strategy 4, although such a destructive and time-consuming strategy is not likely to be practical.     

Effect of ochratoxin A-producing Aspergilli on stilbenic phytoalexin synthesis in grapes

Berries of Vitis vinifera L. cv. Barbera were infected, at veraison and during ripening, by a conidial suspension of A. japonicus, A. ochraceus, A. fumigatus and two isolates of A. carbonarius to control ochratoxin A production and stilbene induced synthesis. The experimental design provided also for intact and punctured berries and incubation temperature of 25 degrees C and 30 degrees C. All the tested fungi, except A. fumigatus, significantly increased trans-resveratrol synthesis over the control, while trans-piceid was not affected; only A. ochraceus significantly elicited the berries to synthesize piceatannol. The two isolates of A. carbonarius produced higher amounts of ochratoxin A than did the other fungi. A positive correlation between ochratoxin A and trans-resveratrol synthesis occurred. trans-Resveratrol and piceatannol showed fungicidal activity against A. carbonarius, being able to completely inhibit fungal growth at a concentration of 300 microg/g and 20 microg/g, respectively.     

Oxidative stress parameters in silver catfish (Rhamdia quelen) juveniles infected with Ichthyophthirius multifiliis and maintained at different levels of water pH

The aim of this study was to determine oxidative stress parameters in the liver, gill and muscle of silver catfish juveniles infected with Ichthyophthirius multifiliis and maintained at pH 5.0 or 7.0 for three days. Juveniles were infected by adding one I. multifiliis-infected juvenile and water containing theronts to tanks. After the appearance of white spots on the skin, infected juveniles exposed to pH 5.0 and 7.0 showed significantly higher thiobarbituric acid reactive substances (TBARS) levels in the liver and gills compared to uninfected juveniles. Liver of infected juveniles exposed to pH 7.0 showed higher catalase (CAT) and lower glutathione-S-transferase (GST) activities, but those maintained at pH 5.0 showed significantly higher GST activity than uninfected juveniles. The gills of infected juveniles showed significantly higher CAT (day two) and GST activity at both pH 5.0 and 7.0 compared to uninfected juveniles. Muscle of infected juveniles showed significantly lower CAT and GST activity and TBARS levels (at day three) when maintained at both pH 5.0 and 7.0 compared to uninfected juveniles. In conclusion, I. multifiliis infection induces liver and gill damage via lipid peroxidation products in silver catfish, but higher antioxidant enzyme activity could indicate a greater degree of protection against this parasite.     

Association of a specific major histocompatibility complex class IIβ single nucleotide polymorphism with resistance to lactococcosis in rainbow trout,Oncorhynchus mykiss (Walbaum)

Major histocompatibility complex (MHC) loci encode glycoproteins that bind to foreign peptides and initiate immune responses through their interaction with T cells. MHC class II molecules are heterodimers consisting of α and β chains encoded by extremely variable genes; variation in exon 2 is responsible for the majority of observed polymorphisms, mostly concentrated in the codons specifying the peptide‐binding region. Lactococcus garvieae is the causative agent of lactococcosis, a warm‐water bacterial infection pathogenic for cultured freshwater and marine fish. It causes considerable economic losses, limiting the profitability and development of fish industries in general and the intensive production of rainbow trout, Oncorhynchus mykiss (Walbaum), in particular. The disease is currently controlled with vaccines and antibiotics; however, vaccines have short‐term efficacy, and increasing concerns regarding antibiotic residues have called for alternative strategies. To explore the involvement of the MHC class II β‐1 domain as a candidate gene for resistance to lactococcosis, we exposed 400 rainbow trout to naturally contaminated water. One single nucleotide polymorphism (SNP) and one haplotype were associated with resistance (P < 0.01). These results are promising for using MHC class IIβ as a molecular marker in breeding rainbow trout resistant to lactococcosis.     

Benthic Fluxes of Dissolved Inorganic Carbon, Nutrients and Oxygen in the Gulf of Trieste (Northern Adriatic)

Benthic fluxes of dissolved inorganic N, Si and P nutrients, alkalinity, dissolved inorganic C (DIC), and O2 from sediments in the Gulf of Trieste (northern Adriatic, Italy) were measured monthly in the period September 1995 - August 1996 using in situ incubated light benthic chambers. The highest efluxes of DIC, NH4+, PO43-, Si(OH)4, and NO3- influxes encountered in late summer - early autumn were the consequence of degradation of benthic microalgae, and in autumn mostly of sedimented phytoplankton. High NO3- efflux was observed in spring. Only NH4+ and Si(OH)4 fluxes were significantly correlated with temperature. This correlation suggests that the rate of downward input and the quality and quantity of sedimentary organic matter (autochthonous and allochthonous) were superimposed on the temperature fluctuations. High DIC, NH4+ and Si(OH)4 effluxes observed in July 1996 were due to the late spring - early summer degradation of sedimentary organic matter produced by benthic microalgae, while the autumn phytoplankton bloom was quickly reflected in enhanced benthic fluxes due to higher temperature. Significant correlations between NH4+, PO43- and Si(OH)4 fluxes suggested their parallel regeneration and utilization at the sediment-water interface. The nutrient fluxes were linked to O2 consumption, suggesting that aerobic oxidation processes were important at the sediment-water interface in the Gulf. The N, P and Si nutrients released from sediment pore waters are probably utilized in benthic microalgal and bottom-water primary production. This indicates that pelagic and benthic communities in the central part of the Gulf of Trieste function relatively independently of each other.     

Estimating heterotrophic and autotrophic soil respiration in a semi-natural forest of Lombardy,Italy

We studied a semi-natural forest in Northern Italy that was set aside more than 50 years ago, in order to better understand the soil carbon cycle and in particular the partitioning of soil respiration between autotrophic and heterotrophic respiration. Here we report on soil organic carbon, root density, and estimates of annual fluxes of soil CO₂ as measured with a mobile chamber system at 16 permanent collars about monthly during the course of a year. We partitioned between autotrophic and heterotrophic respiration by the indirect regression method, which enabled us to obtain the seasonal pattern of single components.The soil pool of organic carbon, with 15.8 (±4.5) kg m^?2, was very high over the entire depth of 45 cm. The annual respiration rates ranged from 0.6 to 6.9 μmol CO₂ m^?2 s^?1 with an average value of 3.4 (±2.3) μmol CO₂ m^?2 s^?1, and a cumulative flux of 1.1 kg C m^?2 yr^?1. The heterotrophic component accounted for 66% of annual CO₂ efflux. Soil temperature largely controlled the heterotrophic respiration (R² = 0.93), while the autotrophic component followed irradiation, pointing to the role of photosynthesis in modulating the annual course of soil respiration.Most studies on soil respiration partitioning indicate autotrophic root respiration as a first control of the spatial variability of the overall respiration, which originates mainly from the uppermost soil layers. Instead, in our forest the spatial variability of soil respiration was mainly linked to soil carbon, and deeper layers seemed to provide a significant contribution to soil respiration, a feature that may be typical for an undisturbed, naturally maturing ecosystem with well developed pedobiological processes and high carbon stocks.