Cytobrushing of the oral mucosa as a possible tool for early detection of testudinid herpesvirus in Horsfield’s tortoises with nonspecific clinical signs

Forty-five Horsfield’s tortoises (Testudo horsfieldii; syn. Agrionemys horfieldii, Russian tortoise) belonging to different owners had decreased appetite and respiratory issues. Twenty-nine tortoises had epiphora, dyspnea, and white necrotic diphtheroid oral plaques (group G1). Ten of the remaining 16 tortoises had serious dehydration, appetite disorder, and depression (G2). The last 6 tortoises had only decreased appetite and moderate conjunctival discharge (G3). During the physical examination of all 45 tortoises, a cytologic sample and an oral swab for herpesvirus and Mycoplasma agassizii PCR testing were taken. In 20 of 29 specimens from G1, in 8 of 16 from G2, and 0 of 6 from G3, the cytologic exam revealed intranuclear acidophilic inclusion bodies, multinucleate cellular syncytia, and further abnormalities caused by herpesviral infection. Moreover, all 45 tested subjects were found to be positive for testudinid herpesvirus 1; 2 were positive for M. agassizii. This prospective study suggests that Horsfield’s tortoises with such signs would benefit from this screening procedure, given that it was effective in a significant proportion of infected and symptomatic animals, and no negative effects were seen.     

Evaluation of ‘di Romagna’ and ‘Frenchgrey’ shallots for quality characters connected to bulb colourand volatile oil content and composition

Local landraces of shallot from the Romagna region(Northern Italy) and North-western France, called Scalogno di Romagna and French grey,respectively, have recently been evaluated for morphological,biochemical and molecular characters. These populations appeared very different from common shallots and onions, so were re-classified as Allium oschaninii O.Fedtsch., whereas almost no variability was observed within these types. Four di Romagna and three French accessions were grown in Italy, and further evaluated for quality characters such asbulb colour and volatile oil content and composition. Colour was measured by means of a tristimulus colorimeter. Volatile oil was extracted by hydrodistillation and its components separated and identified by GC/MS. Colour parameters differentiated thedi Romagna from the French greyaccessions. Essential oil composition differed from what has been reported for most other Alliums, by a lower amount of1-propenyl- substituents. No individual oil component allowed a good differentiation of the two geographic origins.However, the French and di Romagna accessions were perfectly separated by the discriminant analysis of oil composition.Minor 1-propenyl- components seemed more critical for discrimination. The shallot types examined seemed biochemically differentiatedfrom Allium cepa. Variation in the examined quality characters was detected both between the two provenances and among the accessions of common geographic origin.     

Effect of antioxidant protection of must on volatile compounds and aroma shelf life of Falanghina (Vitis vinifera L.) wine

Two vinification methods involving different degrees of antioxidant protection of Falanghina must during prefermentative steps, and referred as HAMP (high antioxidant must protection) and LAMP (low antioxidant must protection), were compared in terms of fermentation performances of four different yeast strains, composition of the volatile fraction of wines at the end of alcoholic fermentation, and shelf life of wines during storage. The use of HAMP technology resulted in wines with lower volatile acidity and higher concentrations of medium-chain fatty acid ethyl esters, acetates, and volatile fatty acids. For two of the four strains a lower concentration of isoamyl alcohol was also observed. HAMP wines also revealed increased shelf life because of the higher concentration of odor active esters at the end of storage and better preservation of varietal aromas.     

Phenolic antioxidants and the protection of low density lipoprotein from peroxynitrite-mediated oxidations at physiologic CO2

Dietary phenolic antioxidants have been shown to prevent LDL modifications mediated by several physiologic oxidants including peroxynitrite. However, more recent data demonstrated that CO(2) affected the fate of peroxynitrite in biological fluids and significantly reduced peroxynitrite scavenging by polyphenols, raising doubts concerning their antioxidant activity. We found that the oxidation of LDL lipids mediated by peroxynitrite decreased in the presence of bicarbonate, while Trp oxidation and 3-nitroTyr formation increased, suggesting a redirection of peroxynitrite reactivity toward the protein moiety. We therefore evaluated the protective activity of some phenolic antioxidants (quercetin, oleuropein, resveratrol, (+)-catechin, (-)-epicatechin, tyrosol, alpha- and gamma-tocopherol, ascorbate) on peroxynitrite-mediated oxidation of LDL aromatic residues. Some of these phenols protected LDL Trp from oxidation better than ascorbate or alpha-tocopherol, although protection at 100 microM did not exceed 30-40%. However, the same phenolic antioxidants were more active in inhibiting 3-nitroTyr formation and those with a catechin structure provided significant protection (IC(50%) 40-50 microM). Red wine, a polyphenol-rich beverage, showed a protective effect comparable to that of the most active phenolic antioxidants. Direct EPR studies showed that bicarbonate significantly increased the peroxynitrite-dependent formation of O-semiquinone radicals in red wine, supporting the hypothesis that polyphenols are efficient scavengers of radicals formed by peroxynitrite/CO(2). Ascorbate was a poor inhibitor of peroxynitrite/CO(2)-induced LDL tyrosine nitration, but the simultaneous addition to the most active polyphenols halved their IC(50%). In conclusion, although cooperation with other antioxidants can further decrease the IC(50%) of polyphenolics, as demonstrated for ascorbate, their antioxidant activity appears to occur at concentrations at least 1 order of magnitude higher than their bioavailability.     

Phenotypic and genetic relationships between yield components in maize

Summary Physiological components of kernel development — LAG period, effective filling period duration (EFPD) and grain filling rate (GFR) — ear moisture release (U), ear size (row number and kernels per row), days from emergence to silking and number of leaves, were examinated on 45 F₁ hybrids (10×10 diallel cross) in order to study their genetic relationships with yield. Combining ability analysis revealed that all trait variability derived mainly from g.c.a. effects. LAG period and EFPD were the traits most affected by genotype-environment interaction.Covariation analysis (path method) based on mean phenotypic values and on g.c.a. effects yielded similar information. It is shown that GFR and EFPD are both related to plant yield, but GFR made the most important contribution. On the contrary, a significant relationship between yield and LAG was not detected. Ear size components were also positively related to yield and had negative effects on GFR. These results indicate that, for our material, the dry matter accumulation rate is the main limiting factor of yield.Considering s.c.a. effects, kernel number per row made the most important contribution.     

Effect of chelating and antioxidant agents on morphology and DNA methylation in freeze-drying rabbit (Oryctolagus cuniculus) spermatozoa

Freeze-drying (FD) has been exhaustively tried in several mammalian species as an alternative technique to sperm cryopreservation, but few studies have been done in rabbits (Oryctolagus cuniculus). The main objective of this study was to compare the protective effect of various antioxidants added to EDTA medium on structural and functional components of FD rabbit spermatozoa and on their status of global DNA methylation. FD media used were composed of basic FD medium (10 mM Tris-HCl buffer and 50 mM NaCl) supplemented with either 50 mM EDTA alone (EDTA) or added with 105 µM of rosmarinic acid (RA, EDTA-RA) or 10 µM of melatonin (MLT, EDTA-MLT). The effect of each medium on the preservation of FD spermatozoon structure was evaluated with light and scanning electron microscopy (SEM). Global DNA methylation was quantified in all FD sperm samples as well as in fresh spermatozoa. Morphologically, fracture points were evidenced in the neck, mid and principal piece of the spermatozoon tail. No differences in spermatozoon fracture points were evidenced among FD treatments: intact spermatozoa were the largest (p < .01) category, whereas the most frequent (p < .01) injury was the neck fracture, resulting in tailless heads. At SEM, the head of spermatozoa showed a well-conserved shape and intact membrane in all treatments. DNA methylation status was the same in all FD treatments. In conclusion, supplementation of EDTA, EDTA-RA and EDTA-MLT during FD preserved rabbit sperm morphological integrity and methylation status as well. Therefore, the difficulty of getting viable offspring using FD semen is likely unrelated to the impact of the lyophilization process on DNA methylation and morphology of lyophilized spermatozoa.     

Hydrolysis of wine aroma precursors during malolactic fermentation with four commercial starter cultures of Oenococcus oeni

The ability of four commercial preparations of Oenococcus oeni lactic acid bacteria (EQ 54, Lalvin OSU, Uvaferm Alpha, and Lalvin 31) to hydrolyze wine aroma precursors was evaluated by measuring the concentration of free and bound aroma compounds at the end of malolactic fermentation carried out in model wines containing a mixture of glycosides extracted from Muscat wine. At pH 3.4 there was a decrease in glycosylated compounds matched by a concomitant increase in free forms in all starter cultures tested. When malolactic fermentation was carried out at pH 3.2, a significant decrease in the ability to hydrolyze aroma precursors was observed for two of the cultures tested (Uvaferm Alpha and Lalvin 31). Large differences in the extent of hydrolysis and in the specificity of this activity toward specific aroma precursors were observed and appeared to be related to the chemical structure of the aglycon as well as to individual characteristics of each starter culture. The amounts of glycosylated aroma compounds released during malolactic fermentation suggest that O. oeni can alter the sensory characteristics of wine through the hydrolysis of aroma precursors.     

Development of a peptide nucleic acid array platform for the detection of genetically modified organisms in food

Two previously developed platforms, a multiplex polymerase chain reaction (PCR) and a peptide nucleic acid (PNA) array, the former allowing for the simultaneous detection of five transgenes and two endogenous controls in food and feed matrices and the latter for the assessment of the identity of amplified PCR products, were combined in order to develop a PNA array device for the screening of genetically modified organisms (GMOs) in food. PNA probes were opportunely designed, synthesized, and deposited on commercial slides. The length of the probes as well as the distance of the probes from the surface were evaluated and found to be critical points. The most suitable probes were found to be 15-mer PNAs linked to the slide surface by means of two 2-(2-aminoethoxy)ethoxyacetic acids as spacers. The device was tested on a model system constituted by flour samples containing a mixture of standards at known concentrations of transgenic material, in particular Roundup Ready soybean and Bt11, Bt176, Mon810, and GA21 maize: The DNA was amplified using the specific multiplex PCR method and tested on the PNA array. The method proposed was found to be able to correctly identify every GMO present in the tested samples.     

Automated on-line solid-phase extraction-liquid chromatography-electrospray tandem mass spectrometry method for the determination of ochratoxin A in wine and beer

An automated on-line solid-phase extraction-liquid chromatography-electrospray tandem mass spectrometry (SPE-LC-ESI-MS/MS) method was developed for the determination of ochratoxin A (OTA) in alcoholic beverages. Mean recoveries for wine and beer were, respectively, 75 and 82%. Detection was achieved in negative ionization with a Q TRAP mass spectrometer operating in multiple-reaction monitoring (MRM) mode or enhanced product ion (EPI) mode, using the third quadrupole as linear ion trap. The MRM mode turned out to be more sensitive; the method allowed accurate determination of OTA in the range of 0.01-25 ng mL(-1) using external calibration. Within-day and between-day relative standard deviation percentages were <6.2 and <9.1%, respectively. In EPI mode, fragmentation spectra at the limit of quantification (0.03 ng mL(-1)) and good linearity could be obtained. Application of the method (MRM mode) to the analysis of several wine and beer samples purchased in local stores revealed OTA levels in the ranges of 0.03-1.44 ng mL(-1) for wines and 0.02-0.14 ng mL(-1) for beers.     

Management practices to control gastrointestinal parasites in dairy and beef goats in Minas Gerais; Brazil

Parasitic infection is recognized worldwide as a limiting factor in the production of goats, and various control methods are used to reduce economic losses, often without considering the epidemiology of the parasites. This has led to the development of highly tolerant parasite populations and the presence of chemical residues in the beef and milk. The objective of this study was to determine the level of knowledge of goat farmers about parasitic diseases and to correlate this with the epidemiology of endoparasites and parasite control practices in goat farms in the state of Minas Gerais, Brazil. The analysis was based on a questionnaire applied by trained veterinarians. The sample was homogeneous throughout the state, covering 18.4% (157/853) of municipalities. Eighty-four dairy goat farms in 81 municipalities and 200 properties with beef goats in 76 municipalities were evaluated. The herd size per goat farm ranged from 4 to 57 (average 24) for beef herds and from 2 to 308 (average 63) for dairy farms. The majority of the beef herd production was extensive and semi-extensive (98.5%), while the dairy herds were maintained under intensive farming (98.8%). The mixed production of goats and sheep was reported by 36.5% of beef goat farmers and by 20.2% of dairy goat farmers. Among the beef goats farms on which the technological level was determined, 2.0% were categorized as having high technological level, 34.5% as medium, and 63.5% as low. Of the 84 dairy farms, 30% operated at a high, 47% at a medium, and 23% at a low technological level. The adoption of practices to reduce parasitism, such as the quarantine of animals, treatment of newly arrived animals, regular cleaning of the floor, and technical assistance, was significantly higher on dairy farms than on beef farms. Although 85.7% of dairy farmers and 83% of beef farmers medicate their animals, the treatments were performed without technical criteria, and deworming intervals ranged from 30 to 120 days or more. The average interval between treatments was significantly longer in dairy goat herds (4.8 months) than in the beef herds (3.6 months). The most commonly used drugs were macrocyclic lactones (37.7% in dairy and 39.5% in beef herds) and benzimidazoles (48.9% in dairy and 31.5% in beef herds). Goat production in Minas Gerais is still in its infancy, and even though using a control program associated with other health practices, producers still rely heavily on chemicals to get satisfactory results.