Assessment of Detoxifying Markers for Florfenicol in Rainbow Trout Liver

Florfenicol (FF) is employed in fish farms to contest or prevent bacterial infections. However, this pharmaceutical may produce reactive oxygen species that may cause biochemical changes in antibiotic-treated fish. The aim of this study was to evaluate the effect of FF on Rainbow Trout Oncorhynchus mykiss treated for 10 d with 7.5 and 15 mg/kg FF followed by a withdrawal period of 5 d. Superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase, glyoxalase I and glyoxalase II, total glutathione, lactic dehydrogenase, and alkaline phosphatase were investigated in the livers of treated and untreated fish. A general impairment of antioxidant enzymes and metabolic indicators was measured in FF-treated Rainbow Trout. Onset of oxidative damage may have occurred during the antibiotic treatment as a consequence of the effect of FF toxicity at mainly the highest dose. Nevertheless, the rise in levels of total glutathione and glutathione S-transferase even after the withdrawal period may shield the antibiotic-mediated oxidative processes.

Received December 22, 2015; accepted May 26, 2016 Published online October 28, 2016     

Pharmacokinetics of IV Ketorolac in Horses Undergoing Orchiectomy

Ketorolac (KET) is a nonsteroidal anti-inflammatory drug for human use, with a potent analgesic activity, that is used in the relief of moderate-to-severe postoperative pain. The pharmacokinetics of KET tromethamine was evaluated after single IV injection at 0.5 mg/kg body weight, after intubation and 10 minutes before surgery, to six Arabian colts undergoing orchiectomy. Intraoperative cardiorespiratory variables were monitored. Blood samples were collected for 36 hours, and serum samples were analyzed by high performance liquid chromotography with ultraviolet-visible detection. During surgery, all monitored physiological parameters were stable. Intermittent positive pressure ventilation, and normocapnia were maintained throughout the procedure in all animals. No adverse effects were observed. The kinetics of KET was described by a two-compartment models, and also a noncompartmental analysis was performed. The distribution and elimination half-lives were t_1/2λ1$t_{1/2{\lambda }_1}$ 0.06 ± 0.02 and t_1/2λ2$t_{1/2{\lambda }_2}$ 0.59 ± 0.21 hours, respectively. Body clearance and mean residence time were 339.99 ± 120.19 mL/h/kg and 0.49 ± 0.22 hours, respectively. The volume of distribution at steady state and volume of distribution based on the terminal phase were 218.83 ± 134.26 mL/kg and 522.5 ± 529.3 mL/kg, respectively. The serum protein binding was 75.8 ± 2.9%. The results indicate that KET at 0.5 mg/kg IV was very rapidly eliminated and thus was likely not effective in the postoperative period. However, further studies including a control group and at higher doses are suggested to investigate the KET kinetics and the analgesic efficacy in horse and define the most appropriate dosage scheme.     

Deltamethrin-impregnated collars for the control of canine leishmaniasis: evaluation of the protective effect and influence on the clinical outcome of Leishmania infection in kennelled stray dogs

A 2-year field study on kennelled stray dogs living in a highly endemic area of leishmaniasis was designed to evaluate whether deltamethrin-impregnated collars (Scalibor) Protector Band) could confer protection against leishmaniasis in this peculiar setting, and to assess differences in clinical outcomes between collared and uncollared dogs. A cohort of 120 clinically healthy and Leishmania-seronegative dogs was enrolled, 50% of which were collared before the 2003 transmission season, and then re-collared before the subsequent season. Collared and uncollared animals were allowed to live with infected dogs in same groups within the kennel. Follow-up included serological (IFAT) assessment twice a year with parasitological Leishmania confirmation, and clinical evaluation performed every 3 months on seroconverted dogs from both groups. Collar losses during the two seasons were high (35%). About 50% of enrolled dogs were lost at follow-up because of death or they were moved to other locations. After the 2003 season, cross-sectional serological examinations tested positive in 5 out of 44 collared animals (11.4%) and in 14 out of 34 controls (41.2%), with 72.3% estimated protection (P<0.005). After the 2004 season, 7/31 seronegative collared dogs seroconverted (22.6%) compared with 7/17 seronegative controls (41.2%), with 45.1% protection (P=0.15). At the end of the study, the cumulative rate of protection was 50.8% (P=0.005). At the clinical evaluation of 21 seroconverted dogs from both groups, canine leishmaniasis signs were significantly more frequent (90% versus 36%, P=0.017) and rapidly progressive in uncollared than in collared dogs. Reasons for such partial clinical protection in collared dogs may be found in the vector anti-feeding effect of protector bands, resulting in a lower number of infectious bites and, probably, in the reduction of antigenic stimuli necessary to shift toward a non-protective immune response.     

In vitro activity of VEGF-E produced by orf virus strains isolated from classical and severe persistent contagious ecthyma

Proliferative orf virus infections in adult sheep have increased in Italy in the past few years: these extreme cases are frequently fatal and difficult to differentiate from other infectious diseases of sheep such as blue tongue. A probable explanation for the proliferative and highly vascularized nature of the lesions was found in the expression of the VEGF-E gene encoded by the orf virus. To investigate a possible role of the viral VEGF in the pathogenesis of severe persistent orf virus lesions, the activity of four VEGF-E variants was compared by an angiogenesis in vitro model. Similar angiogenic activity was found between strains isolated from the classical and the proliferative forms of the disease, even if the latter was able to develop a higher number of vessels during the first 24 h of infection. Our in vitro findings seems to exclude that the VEGF variants encoded by the strain isolated from the atypical form of the disease could be the responsible for the histopathological aspect of the proliferative lesions.     

Comparative analysis of the effects of locally used herbicides and their active ingredients on a wild-type wine Saccharomyces cerevisiae strain

Herbicides are released to the environment with potential ecotoxicological risks for mammals. Yeast is a good model to elucidate toxicity mechanisms. We investigated how three commercial herbicides (Proper Energy, Pointer, and Silglif) and their active ingredients (respectively, fenoxaprop-P-ethyl, tribenuron methyl, and glyphosate) can affect biological activities of an oenological Saccharomyces cerevisiae strain, which may be resident on grape vineyards of the same geographical areas where herbicides are used. The use of commercial grade herbicides employed in Italy allowed us to reproduce the same conditions applied in crops; at the same time, assaying pure single active compounds made it possible to compare the effects obtained with commercial formulations. Interestingly, we found that while pure active compounds affect cell growth and metabolism at a lower extent, commercial preparations have a significant major negative influence on yeast biology.     

Polyphenol content and antioxidative activity in some species of freshly consumed salads

Ten genotypes belonging to Lactuca sativa, Cicorium intybus, Plantago coronopus, Eruca sativa, and Diplotaxis tenuifolia and used in fresh mixed salads were investigated for their polyphenol contents. Flavonoids and hydroxycinnamic acids were characterized by high-performance liquid chromatography (HPLC)/diode array detection/mass spectrometry. Quercetin, kaempferol, luteolin, apigenin, and crysoeriol derivatives were identified; hydroxycinnamic acids were all caffeoyl derivatives. The total polyphenol content was obtained through the Folin-Ciocalteu test and from the HPLC data. The amounts ranged between 0.9 and 4.7 mg/g fresh weight. The antiradical activity was determined by the reaction with the stable DPPH* radical. The Fe2+ chelating activity was determined with a spectrophotometric test. From the complex of data, a quite complete picture of the characteristics of the vegetables emerges. A cultivated C. intybus cultivar exhibited the highest polyphenol content, while a wild C. intybus genotype exhibited the highest antiradical activity. In every case, the characteristics of the different salads as functional foods have been pointed out.     

Evolution of minor polar compounds and antioxidant capacity during storage of bottled extra virgin olive oil

We characterized "Olivastra Seggianese" extra virgin olive oil (EVOO) and evaluated its chemical and sensory characteristics and antioxidant and antiradical activities during storage under novel conditions. Two oils (A and B) were analyzed for the commodity characteristics at blending (t0) and after 9, 12, and 18 months; panel tests were performed and minor polar compounds (MPC) content was assessed at blending (t0) and after 6, 9, 12, and 18 months. Antioxidant and antiradical activities in vitro were evaluated at t0 and after 12 months, by human low density lipoprotein (LDL) and 1,1-diphenyl-2-picrylhydrazil radical (DPPH*) tests. Oil A, which had an initially higher MPC content, possessed "harder" organoleptic characteristics than oil B, which had a lower MPC content and was endowed with a "smoother" taste profile. Statistical analyses showed that secoiridoids, particularly deacetoxy-oleuropein aglycone, should be quantified to evaluate EVOO stability during storage. The antioxidant activity toward human LDL was linked to MPC content and to storage time. The tests on the stable free radical DPPH* confirmed the results on human LDL. We propose this as an additional parameter to evaluate olive oil quality and stability over time.     

Metal ions restore the proteolytic resistance of denatured conglutin gamma, a lupin seed glycoprotein, by promoting its refolding

The susceptibility to trypsin of conglutin gamma, a lupin seed glycoprotein affected by this enzyme only when in a non-native conformation, was used to study the effect of Zn(2+) and other metal ions on the structural dynamics of the protein. When acid-treated trypsin-susceptible conglutin gamma was incubated at neutral pH in the presence of Zn(2+), it became resistant to tryptic attack, contrary to the protein treated in the absence of Zn(2+). The time course of this refolding event has been quantitatively evaluated by SDS-PAGE. Amino acid sequencing of the major polypeptide fragments, produced by trypsin before completion of the refolding process, indicated that only a few cleavable bonds were accessible to the enzyme. This suggested that the presence of metal ions affected the pathway of degradation of the protein, by inducing its folding. Among the other metal ions tested, Ni(2+) also promoted the adoption of a trypsin-resistant conformation of conglutin gamma, whereas Mn(2+) and Ca(2+) had only much lower effects. The relevance of these findings for a deeper understanding of the in vivo degradation of plant food proteins and how it is affected by metal ions are discussed.     

Occurrence of 6-methoxymellein in fresh and processed carrots and relevant effect of storage and processing

The occurrence of 6-methoxymellein (6-MM) in fresh and conventionally processed carrot products (for a total of 176 samples) marketed in European locations and the effect of Alternaria spp. infection and storage conditions on 6-MM accumulation were investigated. 6-MM was found in 78% of tested samples with levels ranging from 0.02 to 76.00 microg/g, with only 1 of 79 fresh carrots exceeding the "just noticeable difference" level for 6-MM. Storage of carrots at 1 degree C was suitable to maintain low levels of 6-MM for a period of at least 17 weeks. No effect of Alternaria spp. infection was observed on 6-MM occurrence. The fate of 6-MM during carrot juice processing was also investigated by using different enzyme formulations for maceration and blanching procedures. Levels of 6-MM in blanched carrots obtained by boiling water or steam treatment were reduced by 69 or 33%, respectively, as compared to fresh carrots. No decrease in 6-MM levels was observed after maceration with pectinolytic enzyme preparations (Rapidase Carrot Juice and Ultrazym AFP-L). A reduction of 6-MM by 85 or 94% was obtained after the entire cycle of carrot juice processing, depending on the blanching procedure used.     

Effects of land use and landscape on spatial distribution and morphological features of gullies in an agropastoral area in Sardinia (Italy)

According to the United Nations Convention to Combat Desertification (UNCCD), soil erosion is one of the main causes of desertification in the Northern Mediterranean. Recently, the role of channelled erosion in sediment delivery has been underlined by several studies. The aim of this paper is to describe a simplified experimental methodology settled up for studying the relationships between gully erosion and land use.The experimental site, which is wide about 720 ha, is located in a hilly area threatened by desertification in Central Eastern Sardinia (Italy). This area is characterised by irregular morphology with steep slopes, where agropastoral activities, particularly tillage with heavy machinery and overgrazing, are causing severe soil erosion and compromising agricultural productivity. Previous studies carried out in the same area highlighted the need to quantify and monitor soil erosion in order to outline management guidelines for farmers. In this study, an integrated methodology that uses aerial photographs and field surveys was applied to inventory the gullies of the area. The degree of activity and morphological characteristics of gullies were assessed in the field by measuring the position and shape of gully heads, and their width and depth. Total gully volume was also estimated. Relationships among distribution, density and morphological characteristics of gullies, and geomorphological features, land use and land use historical dynamics of the area have been studied. The results show the very significant impact of agricultural activities on gully activity and density.