Molecular characterization of equine rotaviruses circulating in Argentinean foals during a 17-year surveillance period (1992-2008)

P[12]G3 and P[12]G14 equine rotaviruses (ERVs) are epidemiologically important in horses. In Argentina, the prevalent ERV strains have been historically P[12]G3. The aim of this study was the detection and characterization of ERV strains circulating in foals in Argentina during a 17-year study (1992-2008). Additionally, the gene sequences of VP7, VP4 and NSP4 encoding genes of representative Argentinean ERV strains were determined and phylogenetic analyses were performed to elucidate the evolutionary relationships of the ERV strains in Argentina. ERVs were detected in 165 (21%) out of 771 diarrheic stool samples, which corresponded to 45 (39%) of 116 outbreaks from the surveyed thoroughbred horse farms. From the positive cases, 51% (n=23) were G3, 33% (n=15) were G14, 4% (n=2) represented a G3+G14 mixed infection and 11% (n=5) of the cases could not be characterized. G3 ERV was detected during the entire period, while G14 ERV was first detected in 2000 and increased its incidence specially in 2006 and 2007. All the analyzed strains belonged to the VP4 P[12] genotype, except for one G3 case which belonged to the P[3] genotype, constituting the first report of a P[3]G3 ERV strain. Phylogenetic analysis of VP7 protein revealed that the G3 Argentinean ERV strains clustered with ERVs from Ireland, while the G14 Argentinean ERV strains formed a distinct cluster within the G14 genotype. The VP4 of the P[12] ERV strains clustered with P[12] strains from Ireland and France. The NSP4 of the Argentinean ERV strains clustered with the NSP4 genotype E12, along with those of guanaco and bovine strains from Argentina, suggesting the a close evolutionary relationship among these Argentinean strains. The results of this study showed changes in the incidence of G3 and G14 during the studied period. The increase in the frequency of G14 ERV, not included in the vaccine, in the second half of the period, may have implications for vaccine design.     

Phylogenetic analysis of the sigma 2 protein gene of turkey reoviruses

The open reading frame of the S3 segment encoding the sigma2 protein of four turkey reovirus field isolates was analyzed for sequence heterogeneity. The turkey reoviruses we present here have a 97% amino acid identity to turkey NC 98. The S3 nucleotide and amino acid sequence similarity was < or =61% and 78%-80%, respectively, when compared to the chicken reovirus isolates. Comparison of amino acid sequences from chickens and turkeys with that of a duck isolate revealed a 53% and 55% similarity, respectively. Phylogenetic analyses, based on both nucleotide and amino acid sequence, resulted in three major groups among the avian reoviruses; these groups were clearly separated by species. The results of this study provide further evidence, based on the deduced sigma2 sequence, that turkey reoviruses form a distinct, separate group relative to chicken and duck isolates. In addition, as a result of the limited sequence identity with their avian counterparts, turkey reoviruses could potentially be considered a separate virus species within subgroup 2 of the Orthoreovirus genus.     

Supplementation of L‐glycine and L‐glutamate to Japanese quails from 01 to 36 days of age using the ideal protein concept

Experiment I: T_1‐1 = basal diet with 25% crude protein (CP) + limiting amino acids (LA); T_1‐2 = 20% CP + LA; T_1‐3 = 20% CP + LA + L‐glycine; T_1‐4 = 20% CP + LA + L‐glutamate; T_1‐5 = 20% CP + LA + L‐glycine + L‐glutamate. Experiment II: T_2‐1 = basal diet with 22% CP + LA; T_2‐2 = 20% CP + LA; T_2‐3 = 17.6% CP + LA + L‐glycine; T_2‐4 = 17.6% CP + LA + L‐glutamate; T_2‐5 = 17.6% CP + LA + L‐glycine + L‐glutamate. The reduction of dietary protein based on the concept of ideal protein decreases nitrogen excretion in quails when L‐glycine is added to the diets. Quails fed diets supplemented with L‐glutamate as the non‐specific nitrogen source equivalent to the nitrogen level of the control diet had increased nitrogen excretion. However, quails had reduced nitrogen excretion in both experiments when L‐glycine was added to diets with L‐glutamate. Carcass fat was increased by reducing dietary protein, but fat deposition was reduced by adding L‐glutamate and L‐glycine, or both. The dietary addition of L‐glutamate and L‐glycine in quails based on the ideal protein concept is not necessary (Exp. I). Although the total nitrogen, electrolytic balance and glycine level were adjusted in diets, quails had decreased performance. Therefore, other hypotheses besides protein reduction need to be studied (Exp. II). Protein reduction with supplementation of only limiting essential amino acids does not affect quail performance. Dietary addition of L‐glycine reduces nitrogen excretion.     

Cloning and expression of pigeon IFN-γ gene

This is the first paper describing the cloning of pigeon IFN-γ gene (PiIFN-γ) and the analysis of the in vitro expressed recombinant protein. The PiIFN-γ gene was identified by RT-PCR as a 498 bp, fragment coding for a precursor protein of 165 amino acids instead of 164 amino acids, as observed in the other avian species. The recombinant protein was expressed in vitro by an eukaryotic system and the biological properties of the cytokine were tested using a chicken macrophage cell line. The high degree of amino acid and nucleotide identity, shared with the ChIFN-γ, and the fact that the pigeon protein was functional on chicken cells, indicates a cross-reactivity between pigeon and chicken IFN-γ. The detection of the PiIFN-γ could represent an useful instrument in understanding the role played by this cytokine in immune response related to vaccinations and infectious diseases in the pigeon.     

Acute phase proteins and biomarkers of oxidative status in feline spontaneous malignant mammary tumours

Acute phase proteins (APP) and biomarkers of oxidative status change in human and canine mammary tumours, however, they have not been studied in feline mammary tumours. The aims of this study were to investigate the APP and antioxidant responses in feline malignant mammary tumours, to evaluate their relation with tumour features, and to assess their prognostic value. Serum amyloid A (SAA), haptoglobin (Hp), albumin, butyrylcholinesterase (BChE), insulin‐like growth factor1 (IGF1), paraoxonase1 (PON1), total serum thiols (Thiol), glutathione peroxidase (GPox) and total antioxidant capacity determined by different assays, including trolox equivalent antioxidant capacity assessed by two different methodologies (TEAC1/2), ferric reducing ability of plasma (FRAP), and cupric reducing antioxidant capacity (CUPRAC), were determined in serum of 50 queens with spontaneous mammary carcinomas and of 12 healthy female cats. At diagnosis, diseased queens presented significantly higher SAA and Hp, and lower albumin, BChE, GPox, TEAC1, TEAC2 and CUPRAC than controls. Different tumour features influenced concentrations of APP and antioxidants. Increases in serum Hp, and decreases in albumin, Thiol and FRAP were significantly associated with neoplastic vascular emboli, metastasis in regional lymph nodes and/or in distant organs. Distant metastasis development during the course of the disease was associated with increases in SAA and TEAC1. At diagnosis, decreased albumin was associated with a longer survival, and BChE <1.15 μmoL/mL.minute was associated with a shorter survival time on multivariate analysis. Feline malignant mammary tumours are associated with an APP response and oxidative stress, and different tumour features influence the inflammatory response and the oxidative damage. Furthermore, some of these analytes proved to have prognostic value.     

Transvenous coil embolisation for the treatment of single congenital portosystemic shunts in six dogs

This article describes the treatment of single congenital portosystemic shunts (CPSs) (intrahepatic and extrahepatic) using an interventional radiology technique involving embolisation of anomalous vessels with percutaneous coils. Briefly, a multipurpose catheter was introduced into the caudal vena cava and then into the portosystemic shunt. An autoexpandable stent was placed in the caudal vena cava, next to the shunt, in order to avoid coil migrations, and a cobra-like vascular catheter was used to pass through the stent and to place the coils in the shunt. This technique was used for treatment of CPS in six dogs. The results indicate that percutaneous embolisation of a CPS using coils, a less invasive technique than the traditional surgical technique, may result in complete closure of the anomalous vessel without development of portal hypertension.     

Arsenic Mining Waste in the Catchment Area of the Madrid Detrital Aquifer (Spain)

In recent years, elevated arsenic concentrations in groundwater used for drinking water supplies have been recognised in the Madrid Tertiary detrital aquifer. Although only natural causes have been suggested as the source of arsenic, this study aims to highlight that the anthropogenic contribution cannot be disregarded. During the sub-catchment??s areas sampling, we found many geographical sites where natural arsenopyrite [FeAsS] originally encapsulated in pegmatite bodies and quartz veins, was artificially outcropped and dumped out, since mining wastes were scattered and exposed to weathering. Several mineral and ground specimens were collected to analyse its mineralogical and chemical composition by X-Ray Diffraction (XRD) and X-Ray Fluorescence (XRF) spectrometry and by Environmental Scanning Electron Microscope (ESEM). Both, the abundant existence of secondary phases, such as scorodite [FeAsO₄?2H₂O] and jarosite [KFe₃(SO₄)₂(OH)₆], much more soluble than arsenopyrite, and the lixiviation experiments of arsenopyrite in acidic media to simulate acid mine drainage (AMD) conditions, usually found in old mining districts, point to a potential risk of arsenic contamination of surface water bodies, which operate as recharged waters of the aquifer in the studied area. The elemental determination of heavy metals present in ground samples by XRF analyses, reaching up to 1,173 mg kg^?C1 of copper, 347 mg kg^?C1 of lead and 113,702 mg kg^?C1 of arsenic; and the physicochemical and arsenic fractionation studies of soil samples, led us to classify the soil as Spolic Technosol (Toxic). The contamination of the area due to old mining activities could release arsenic to Madrid water supplies; accordingly, additional decontamination studies should be performed.     

Secoiridoids, tocopherols, and antioxidant activity of monovarietal extra virgin olive oils extracted from destoned fruits

The effect of olive stone removal before processing on the degradation level, secoiridoid and tocopherol contents, and antioxidant activity of monovarietal extra virgin olive oils (EVOOs) was studied. EVOOs were extracted from olives of the Leccino, Moraiolo, Frantoio, Pendolino, Taggiasca, and Colombaia varieties both in the presence and in the absence of the stones. The degradation level of EVOOs was evaluated by acidity, peroxide number, and spectroscopic indices K(232) and K(270), according to EU regulation. The secoiridoid compounds typical of EVOO, namely, the oleuropein and ligstroside derivatives, hydroxytyrosol, tyrosol, and tocopherols were analyzed by HPLC. The antioxidant activity was evaluated by the xanthine oxidase/xanthine system, generating superoxide radical and hydrogen peroxide, and by the 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl test. Results showed that EVOOs obtained from both stoned and destoned olives had a very low degradation level, which was not affected by destoning. Destoning lowered slightly the alpha-tocopherol content in EVOOs but increased the total secoiridoid content and the antioxidant activity of EVOOs (up to 3.5-fold). However, these effects were variety-dependent and negligible in some conditions. It was concluded that a better knowledge of the reactions occurring during olive processing, and particularly on the involvement of endogenous pulp and stone enzymes, is essential to predict the effect of destoning on EVOO quality.     

Collection and morphological characterization of Panicum coloratum L. in Argentina

Panicum coloratum (Poaceae) is a perennial grass native to South Africa, distributed all over the world, but poorly adopted by ranchers in Argentina. Adapted to a wide range of soil and climatic conditions, it constitutes a good forage resource for zones with edaphic and climatic restrictions, where cattle rising activities are here displaced. The high polymorphism in the species has made it difficult to delineate sub specific taxonomic categories. It has been classified in botanical varieties, types and/or cultivars. In order to document the morphological variability in P. coloratum and to investigate the differences between var. makarikariense and var. coloratum, a collection was gathered in INTA Rafaela composed by fourteen accessions from different provinces of Argentina: six accessions of var. makarikariense and eight from var. coloratum. The evaluation of 152 individuals gives evidence of a wide morphological variation in the collection in traits related to forage and caryopses production; seven out of nine traits showed significant differences between var. makarikariense and var. coloratum. Based on the study of local type specimens, a morphological key to distinguish between P. coloratum varieties is provided. These results report new information about the diversity in P. coloratum in Argentina and provide valuable insights for future use of the collection as the base material of a breeding program.