Follicular fluid concentrations of free insulin-like growth factor (IGF)-I during follicular development in mares

The objective of the present study was to evaluate changes in concentrations of free insulin-like growth factor (IGF)-I in follicular fluid (FFL) during follicle development in the mare. Mares (n = 14) were classified as either in the follicular phase (n = 8) or luteal phase (n = 6). Follicles (n = 92) were categorized as small (6–15 mm; n = 54), medium (16–25 mm; n = 23) or large (>25 mm; n = 15) and FFL was collected. Free IGF-I levels in FFL in large follicles of follicular phase mares were greater (P < 0.05) than in large follicles of luteal phase mares and small or medium follicles of luteal and follicular phase mares. Free IGF-I concentrations were greater (P < 0.05) in large follicles of luteal phase mares than small but not medium follicles of luteal phase mares. FFL ratio of estradiol:progesterone paralleled changes in free IGF-I. Free IGF-I concentrations were negatively correlated (P < 0.05) with insulin-like growth factor binding protein (IGFBP)-2, -4 and -5 but not IGFBP-3 levels. In addition, free IGF-I concentrations in FFL were positively correlated (P < 0.01) with FFL estradiol, progesterone, androstenedione, estradiol:progesterone ratio, total IGF-I and total IGF-II. We conclude that increases in intrafollicular levels of bioavailable (free) IGF-I are associated with increased steroidogenesis in developing mare follicles.     

Differentiation between canine cutaneous and visceral leishmaniasis by the detection of immunoglobulin G specific for Leishmania (Viannia) braziliensis and Leishmania (Leishmania) chagasi antigens using flow cytometry

Flow cytometry employing Leishmania (L.) chagasi (Lc) and L. (Viannia) braziliensis (Lb) antigen was used to establish the differential diagnosis between visceral (VL) and cutaneous leishmaniasis (CL) in dogs. Flow cytometry permitted the detection of Leishmania-specific immunoglobulin G in sera from 19 dogs: nine with CL and 10 with VL. A significant difference in the percentage of positive staining was observed in sera from dogs with CL between the homologous antigen (69% for Lb) and the heterologous antigen (42% for Lc). However, this difference was not significant in sera from dogs with VL (61% for Lb and 73% for Lc). No significant staining was observed in control sera (0.6% for Lb and 0.4% for Lc) consisting of samples from healthy dogs, or in the group with sporotrichosis (1.8% for Lb and 1.5% for Lc), a differential diagnosis of CL. The results suggest that flow cytometry might be useful for the differentiation between CL and VL in dogs, with practical applications in areas where the two infections overlap.     

Molecular evolution of tomato black ring virus and de novo generation of a new type of defective RNAs during long-term passaging in different hosts

Tomato black ring virus (TBRV) is a worldwide-distributed RNA virus infecting a wide range of different host plants, including crop species, trees, shrubs, and weeds. Here, we investigated the molecular evolution of TBRV and its adaptability to different plant species. The TBRV-Pi isolate was used to generate five independent evolution lineages serially passaged in either quinoa, tobacco, or tomato plants. After 15 passages, the genetic variability present in all the lineages was characterized for the movement (MP) and coat (CP) coding cistrons. We addressed two main questions: to what extent does the amount of genetic variability in the TBRV genome depend on the host species, and are there host species-specific adaptive mutations? Overall, 201 different nucleotide substitutions emerged during the evolution experiment, with some of them appearing multiple times in different lineages; two of them (one in CP and one in MP) were unique for a particular host plant. We have shown that the degree of genetic differentiation depends on the host species in which the virus evolved, and that positive selection is operating upon certain residues, particularly in CP. Moreover, we have characterized new types of defective RNAs that arose during the TBRV-Pi evolution in tobacco. Furthermore, this is the first report of a defective RNA from the RNA2 of TBRV.     

Optimisation of Healthy-Lipid Content and Oxidative Stability during Oil Extraction from Squid (Illex argentinus) Viscera by Green Processing

Green extraction was applied to Argentinean shortfin squid (Illex argentinus) viscera, consisting of a wet pressing method including a drying step, mechanic pressing, centrifugation of the resulting slurry, and oil collection. To maximise the oil yield and ω3 fatty acid content and to minimise the oil damage degree, a response surface methodology (RSM) design was developed focused on the drying temperature (45–85 °C) and time (30–90 min). In general, an increase of the drying time and temperature provided an increase in the lipid yield recovery from the viscera. The strongest drying conditions showed a higher recovery than 50% when compared with the traditional chemical method. The docosahexaenoic and eicosapentaenoic acid contents in the extracted oil revealed scarce dependence on drying conditions, showing valuable ranges (149.2–166.5 and 88.7–102.4 g·kg⁻¹ oil, respectively). Furthermore, the values of free fatty acids, peroxides, conjugated dienes, and ω3/ω6 ratio did not show extensive differences by comparing oils obtained from the different drying conditions. Contrary, a polyene index (PI) decrease was detected with increasing drying time and temperature. The RSM analysis indicated that optimised drying time (41.3 min) and temperature (85 °C) conditions would lead to 74.73 g·kg⁻¹ (oil yield), 1.87 (PI), and 6.72 (peroxide value) scores, with a 0.67 desirability value.     

Anti-oxidant status in embryonic,post-hatch and larval stages of Asian seabass (<Emphasis Type="Italic">Lates calcarifer</Emphasis>)

The concentrations of anti-oxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and selenium-dependent glutathione peroxidase (SeGPx), and low molecular weight free-radical scavengers such as reduced glutathione (GSH) and ascorbic acid (vitamin C) were evaluated during the period from gastrulation (GS) to 25 days post-hatch (dph) in the larvae of Asian Seabass, Lates calcarifer. Oxidative damage due to lipid peroxidation (LPO) was also assessed, by evaluation of the formation of malondialdehyde (MDA). All the three anti-oxidant enzymes, SOD, CAT and GPx, showed high activities during gastrulation, suggesting an increased metabolic rate during the period of embryonic development. Though the SOD activity apparently decreased progressively during 3–20 dph of larval development, the difference was not significant. CAT showed high activity during gastrulation and remained constant up to 3 dph, suggesting an increased need to metabolise hydrogen peroxide (H₂O₂) and organic peroxides. In contrast, SeGPx activity increased progressively from 5 dph to 25 dph during larval development, indicating an increased need to detoxify lipid peroxides. This is evident from the observation of increased lipid peroxidation from 10 dph to 25 dph during larval development. GSH levels were low at gastrulation, indicating increased metabolic rate and formation of lipid radicals during this period, corresponding to the decrease in the level of ascorbic acid, which is consumed for regeneration of GSH.     

Regulated deficit irrigation in green bean and watermelon greenhouse crops

Mediterranean greenhouse growers of watermelon and green bean crops tend to reduce slightly the soil water availability during the flowering phase to enhance the fruit number and yield, but without measuring the soil or the plant water status. This deficit irrigation strategy (RDI) was studied on two representative growth cycles of green bean and one of watermelon. In each case, a well-watered crop acted as control.In the three well-watered vegetable crops, soil water matric potential (SMP) values were between −20 and −30 kPa throughout most of the respective growth cycles. These values avoid water deficits in Mediterranean greenhouse vegetable crops. The watermelon under RDI presented similar SMP to the well-watered crop, except during the flowering period when it reached values of −50 to −60 kPa, which are similar to, or slightly lower than, those recommended to prevent water deficits for cucurbitaceae crops. The autumn–winter and spring cycles of green bean under RDI presented progressively lower SMP values from the vegetative phase to the first fruit setting than the well-watered crops, reaching minimum SMP values of around −55 kPa for the autumn–winter cycle and of −75 kPa for the spring one. These minimum SMP values are similar for the autumn–winter cycle and lower for the spring cycle than those recommended to avoid water deficits in green bean crops grown in medium-fine textured soils. Overall, mild water deficits during flowering of watermelon and green bean crops grown in Mediterranean greenhouses did not improve the final fruit number or yield. In the two spring cycles (watermelon and green bean) the RDI strategy reduced the aboveground biomass and yield, whereas in the autumn–winter green bean cycle the RDI strategy reduced the vegetative biomass but did not affect yield. SMP threshold values can, however, be used by growers as a tool for controlling the equilibrium between the vegetative and reproductive growth of greenhouse soil-grown crops.     

Evaluation of the Influence of Natural and Antrhopogenic Processes on Water Quality in Karstic Region

This paper presents a comprehensive study on the quality of surface and groundwater in an environmentally sensitive karstic watershed strongly pressed by urban growth. The objective of the study was to assess the integrated effects of human activities and natural characteristics of karstic environments on the quality of surface and groundwater using multivariate statistical techniques. Data from 18 physical, chemical, and microbiological water quality variables obtained throughout a hydrological year were analyzed using principal components analysis (PCA) and cluster analysis. The PCA was carried out individually for surface water and groundwater. Our goal was to study the behavior of the water quality variables at each of these systems, as well as to infer the importance of these variables on the dynamics of the water resources in the region. Our results for surface water showed that the 18 original variables could be reduced to five principal components which together accounted for 69% of the total variation in the data, whereas for groundwater, 70% of the total variation in the data was explained by five principal components. In order to evaluate the impacts of anthropic activities on the water quality at the monitoring locations, the cluster analysis was applied to the ten sampling points. The analysis identified five clusters, two of which are formed by points with low contamination levels and three where the anthropic interference is noticeable. The results suggest the existence of specific contamination sources in many points, including in the groundwater, and highlight the natural vulnerability of the karstic environments.     

Prevalence and genetic characterization of Hepatitis E virus in paired samples of feces and serum from naturally infected pigs

This study describes the distribution of Hepatitis E virus (HEV) in a naturally infected swine population and the genetic relatedness of HEV strains on swine farms in Spain. Of fecal and serum samples collected from 131 pigs and manure-ditch samples collected from 17 farms, HEV was detected in 16%, 14%, and 59%, respectively, for an overall prevalence rate of 23%. The maximum prevalence rates for feces and serum were in pigs 5 to 12 wk old. A high prevalence of the virus in feces (18%) was observed in sows. Gene sequencing was performed on 6 strains from feces, serum, and manure ditch: the nucleotide identities varied from 81.5% to 99% when compared with those of other strains of genotype 3 isolated from swine. This is the first study in Europe to show the variation in virus distribution by age in feces and serum in a naturally infected swine population.     

Mixed germ cell tumor in the eye of a dog.

A 3-year-old female neutered Staffordshire Bull Terrier presented with a mixed germ cell tumor involving the base of the iris and the ciliary body of the right eye. The tumor mass was composed primarily of packeted vacuolated, polygonal (hepatoid) cells and small round cells; epithelial cells lining tubuloacinar structures were a less prominent component. The hepatoid and round cells stained positively for alpha-fetoprotein and cytokeratin. The epithelial cells stained positively for cytokeratin only, and some contained cytoplasmic mucin droplets. The polygonal cells were interpreted as a hepatoid variant of yolk sac tumor, and the epithelial cells were considered a teratomatous component. Trabeculae of bone were observed within the mass and may have been metaplastic or a teratomatous element. Extragonadal germ cell tumors are rare in dogs and have previously been reported only in the suprasellar region. This is the first report of this tumor type in the eye of a nonhuman species.