Specific polymerase chain reaction for differential diagnosis of Dirofilaria immitis and Dipetalonema reconditum using primers derived from internal transcribed spacer region 2 (ITS2)

Both Dirofilaria immiti and Dipetalonema reconditum may be found in blood of infected dogs but it is not easy to distinguish D. immitis from D. reconditum in morphology. We cloned and sequenced the contiguous internal transcribed spacer (ITS) region, ITS1-5.8S-ITS2, of these two different parasites and published on GenBank as AF217800 for D. immiti and AF217801 for D. reconditum in this study. We designed two pairs of specific primers derived from ITS2 being used for polymerase chain reaction (PCR). The amplicons of ITS2 from D. immiti and D. reconditum are 302 and 348bp, respectively. Moreover, the limitation for amplifying ITS2 gene using this PCR demonstrated that 1 x 10(-2) microfilaria of each species of parasite smashed or even with mixed samples could be detected and the PCR products were predicted as the same as that described above. Thus, D. immiti and D. reconditum could be differentially diagnosed by this specific PCR. Seventeen clinical cases were evaluated and all of them were correctly identified. In this study, ITS1-5.8S-ITS2 of D. immiti or D. reconditum were the first time sequenced and analyzed. No significant similarity of ITS1 and ITS2 between D. immiti and D. reconditum could be observed.     

Influence of diet and feeding strategy on the performance of nitrifying trickling filter,oxygen consumption and ammonia excretion of gilthead sea bream (Sparus aurata) raised in recirculating aquaculture systems

Gilthead sea bream (Sparus aurata) was raised in six individual recirculating aquaculture systems (RAS) whose biofilters’ performance was analyzed. Fish were fed with three different diets (a control diet, a fishmeal-based diet (FM), and a plant meal-based diet (VM)) and with three different feeding strategies (manual feeding to apparent satiation, automatic feeding with restricted ration, and auto-demand feeding). For every combination of diet and feeding strategy, the mean oxygen consumption, ammonia excretion, and ammonia removal rate were determined. Fish fed with the VM diet consumed the most oxygen (20.06?±?1.80 gO₂ consumed kg^?1 day^?1). There were significant differences in ammonia excretion depending on the protein content and protein efficiency of the diet, as well as depending on feeding strategy, which in turn affected ammonia removal rates. Fish fed by auto-demand feeders led to the highest mean ammonia removal rate (0.10 gN-TAN removed m^?2 biofiltration area day^?1), while not leading to peaks of high ammonia concentration in water, which preserve fish welfare and growth.

     

Vermicomposting leachate as liquid fertilizer for the cultivation of sugarcane (Saccharum sp.)

In vermicomposting, the main product is the worm casts, but a leachate is generated that contains large amounts of plant nutrients. This leachate is normally diluted to avoid plant damage. We investigated how dilution of vermicompost leachate combined with different concentrations of nitrogen (N) - phosphorus (P) - potassium (K) triple 17 fertilizer, and polyoxyethylene tridecyl alcohol as dispersant and polyethylene nonylphenol as adherent to increase efficiency of fertilizer uptake, affected sugarcane plant development. The vermicomposting leachate with pH 7.8 and electrolytic conductivity 2.6 dS m^?1, contained 834 mg potassium (K) l^?1, 247 mg nitrate (NO₃^?) l^?1 and 168 mg phosphate (PO₄^3?) l^?1, was free of pathogens and resulted in a 65% germination index. Vermicompost leachate did not inhibit sugarcane growth and mixed with 170 g l^?1 NPK triple 17 fertilizer resulted in the best plant development. No dispersant or adherent was required to improve plant height and stem development.     

Influence of Salinity in the Bioavailability of Zn in Sediments of the Gulf of Cádiz (Spain)

This study assesses the effect of salinity in bioavailability and toxicity of Zn by means of laboratory bioassays by observing contamination in both sediment and water, accumulation of Zn in biological tissues, and histopathological damage in the gills and guts tissues of Ruditapes philippinarum clams, which were exposed to different types of sediments from the Gulf of Cádiz (SW Spain) as well as two dilutions of toxic mud coming from an accidental mining spill. With this objective, the coefficients of distribution (K _D) for Zn between overlying water and sediments were calculated, the histopathological frequencies in the tissues of the gills and guts of clams were determined, and the biota-sediment bioaccumulation factors as well as the bioaccumulation factors were quantified in the different stations. Results showed that the greatest histopathological damages appeared when the salinity values decreased. Statistical results showed that salinity was inversely correlated with histopathological damage (p?<?0.01) for the lesion index for gills. The most outstanding results were observed in the two dilutions of toxic mud (0.3% and 7.9%) at a salinity value of 10. Salinity was inversely correlated with the concentration of Zn in biological tissues (p?<?0.05) and inversely correlated with the concentration of Zn in water and sediment. Zn mobilization to the overlying water is produced when salinity values decrease.     

Classification of wine and alcohol vinegar samples based on near-infrared spectroscopy. Feasibility study on the detection of adulterated vinegar samples

Near-infrared (NIR) spectroscopy was used to discriminate between wine vinegar (red or white) and alcohol vinegar. One orthogonal signal correction method (OSC) was applied on a set of 73 vinegar NIR spectra from both origins and artificial blends made in the laboratory in order to remove information unrelated to a specific chemical response (tartaric acid), which was selected due to its high discriminant ability to differentiate between wine vinegar and alcohol vinegar samples. These corrected NIR spectra, as well as raw NIR spectra and 14 physicochemical variables, were used to develop separate classification models using the potential functions method as a class-modeling technique. The aforementioned models were compared to evaluate the suitability of NIR spectroscopy as a rapid method for discriminating between vinegar origins. The transformation of vinegar NIR spectra by means of an orthogonal signal correction method prompted a notable improvement in the specificity of the constructed classification models. The classification model developed was then applied to artificial vinegar blends made in the laboratory to test its capacity to recognize adulterated vinegar samples.     

Histological alterations in gills of shrimp Litopenaeus vannamei in low‐salinity waters under different stocking densities: Potential relationship with nitrogen compounds

Two experimental modules with different stocking densities (M1 = 70 and M2 = 120 shrimp /m²) were examined weekly over a culture cycle in tanks with low‐salinity water (1.9 g/L) and zero water exchange. Results showed survival rates of 87.7 and 11.9% in M1 and M2, respectively. Water temperature, pH, dissolved oxygen, electrical conductivity and chlorophyll a were not significantly (p > .05) different between modules. In contrast, the concentrations of nitrogen compounds were significantly (p < .05) different between modules, except nitrite‐N (M2 were 2.31 ± 1.38 mg/L N‐TAN, 0.18 ± 0.49 mg/L N‐NO₂^? and 6.83 ± 6.52 mg/L N‐NO₃^?; in M1: 0.97 ± 0.73 mg/L N‐TAN, 0.05 ± 0.21 mg/L N‐NO₂^? and 0.63 ± 0.70 mg/L N‐NO₃^?). When waters of both modules reached higher levels of ammonia and nitrite, histological alterations were observed in gills. The histological alterations index (HAI) was higher in M2 (5‐112) than in M1 (2‐22).     

Post-fire production of mushrooms in Pinus pinaster forests using classificatory models

This study was aimed at describing post-fire mushroom production in a Mediterranean ecosystem dominated by Pinus pinaster Ait. in the northwest of Spain and assessing the results by classificatory models. During the autumn periods of 2003–2006, fruit bodies from 115 fungal taxa were collected in burned and unburned areas and were further grouped into the following categories: saprotrophic/mycorrhizal; and edible/non-edible. After wildfires, a significant reduction in the number of fungal species and fruit body biomass production was observed. Based on this relevant information, the first simple classificatory model for this aim is provided. Nine alternative models based on classifications according to combinations of edibility and functional groups were fitted, and four fruiting body biomass production classes were defined as possible responses. As explanatory factors, time after fire and climatic variables significantly related to fruit body production were included. The best predictive results were obtained for edible and edible-mycorrhizal models, for which the correct classification rate of production classes was between 92 and 85 %. Moreover, the models obtained were applied to analyse the effect of time after fire on fungal production. Mycorrhizal and edible fungal production after fire was classified into the lowest class, whereas saprotrophic and non-edible species followed a contrary trend. The classificatory models can be useful to optimise management and harvest of these increasingly appreciated non-timber forest resources.     

Characterization of Collagen from Different Discarded Fish Species of the West Coast of the Iberian Peninsula

ABSTRACT

Skin collagen of six discarded fish species was analyzed and compared. Acid soluble collagen (ASC) was extracted; a characteristic sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profile for type I collagen was obtained, except for Chimaera mostrosa. Contents of collagen calculated from HPro (31.85% average) were higher than those determined from ASC extracts (17.75% average), with Galeus spp. being the species with the higher percentage. Amino acid analysis revealed the typical composition of collagen, with very few differences among species. Specific profiles were obtained after protease digestion. Denaturation temperature of ASC correlated well with imino and hydroxyproline contents.

Results demonstrate the feasibility of using the obtained collagens in different industrial applications.     

Dual-label time-resolved fluoroimmunoassay for simultaneous quantification of haptoglobin and C-reactive protein in meat juice from pigs

A new method was developed to simultaneously measure 2 acute-phase proteins (APPs) by time-resolved immunofluorometry. The assay, based on double-label quantification of haptoglobin (Hp) and C-reactive protein (CRP) in meat juice samples from pigs, was constructed by use of a combination of europium and samarium chelate lanthanides as labels. Meat juice samples from 154 pigs were used for analytic and clinical validation of the assay through determination of precision, accuracy, limit of detection, and quantification. The analytic performance of the assay was satisfactory, with good intra-assay and interassay precision and accuracy. The levels of Hp and CRP were increased in the meat juice samples of diseased animals compared with healthy ones. According to the results, higher sensitivity could be achieved if the cut-off values of both proteins were taken into account for clinical relevance rather than used individually. Since the dual assay saved both time and sample, it could be used as a rapid and sensitive screening test in porcine production.