Modulation of tobacco bacterial disease resistance using cytosolic ascorbate peroxidase and Cu,Zn‐superoxide dismutase

The effect of over‐expression in tobacco plants of cytosolic Cu,Zn‐superoxide dismutase (cytsod) and ascorbate peroxidase (cytapx) alone, or in combination, against bacterial wildfire and crown gall diseases, caused by Pseudomonas syringae pv. tabaci and Agrobacterium tumefaciens, respectively, was investigated. Disease tolerance was observed in all the transgenic lines against the two causal agents, with various levels of resistance, with the double transformants (lines 35 and 39) the most resistant against bacterial wild fire. In the case of P. syringae pv. tabaci, disease tolerance and symptom decrease was associated with a lower bacterial population and a higher level of several antioxidant defence enzymes. Transgenic lines also exhibited an enhanced tolerance against A. tumefaciens, with the transgenic line harbouring cytapx (line 51) the most resistant to crown gall disease. However, this was only observed with strain C58 among the three pathogenic strains tested. These results suggest that cytosolic antioxidant defences have a role in increasing tolerance to the oxidative stress caused by some bacterial pathogens, and resistance of these tobacco lines to wildfire disease seems to be independent of tissue necrosis.     

Prevalence of infectious diseases in feral cats in Northern Florida

Objectives of this study were to determine prevalence of infection in feral cats in Northern Florida with a select group of infectious organisms and to determine risk factors for infection. Blood samples or sera from 553 cats were tested with a panel of antibody, antigen or PCR assays. Male cats were at higher risk for FIV, Mycoplasma haemofelis, and M. haemominutum. Infection with either FeLV or FIV was associated with increased risk for coinfection with the other retrovirus, M. haemofelis, or M. haemominutum. Bartonella henselae had the highest prevalence and was the only organism that did not have any associated risk for coinfection with other organisms. Feral cats in this study had similar or lower prevalence rates of infections than those published for pet cats in the United States. Thus, feral cats assessed in this study appear to be of no greater risk to human beings or other cats than pet cats.     

Sustained availability of trimethoprim in drinking water to achieve higher plasma sulphonamide-trimethoprim antibacterial activity in broilers

(1) In order to make trimethoprim (TMP) available to broilers throughout the day, a sustained release formulation (SRF) of the drug in the form of granules was added to the water tank that supplies drinking water. (2) Broilers were initially dosed with sulphachloropiridazine-TMP (SCP-TMP 5:1) and then further medicated throughout the day, achieving in the end a dose of 30 mg/kg each of SCP and TMP (group A). Group B received a preparation with the same dose of SCP and TMP (1:1) as group A, but administered as a single dose without the SRF of TMP. Group C received the customary SCP-TMP 5:1 preparation (30 and 6 mg/kg, respectively). Water tanks were completely consumed in 3 to 4 h. (3) Broilers were bled at different times and concentration of antibacterial activity in serum determined by correlating the composite antibacterial activity of SCP and TMP with actual concentrations of these drugs by means of a microbiological agar diffusion assay. (4) Time vs serum concentrations of activity were higher in group B; the increments in the maximum serum concentration for group B over groups A and C being 39 and 67%, respectively. (5) However, the sustained concentration of activity over time, measured as the area under the cu)rve, was highest in group A. Group B had higher values for area under the curve than group C. (6) An additional dose of TMP to achieve 30 mg/kg of both SCP and TMP improves the serum concentration of this combination over the customary 5:1 proportion. The best values for sustaining antibacterial activity were obtained using a 1:1 ratio as in group A. The use of a SRF as in group A may translate into better clinical results.     

Pharmacokinetics and residues of a new oral amoxicillin formulation in piglets: a preliminary study

The pharmacokinetics of amoxicillin (Amx) were determined in pigs following intravenous (IV) administration of a single dose of 15 mg/kg and a single dose of 15 mg/kg of a new oral formulation (Amx-FP containing 10% amoxicillin). Residue studies were performed to determine residues in edible tissues of healthy pigs after chronic oral administration of Amx-FP at a daily dose of 15 mg/kg for five consecutive days. After IV administration, the plasma concentration was characteristic of a two-compartment open model. The main pharmacokinetic variables were: t(1/2lambda(n)), MRT=90.1 min, V(darea)=0.81 L/kg and Cl(b)=3.9 mL/kg/min. After single oral administration the main pharmacokinetic variables were: C(max)=758 mug/L, t(max)=347 min and Cl(b/f)=3.7 mL/kg/min for Amx-FP. The oral bioavailability (F) was calculated at 11% for Amx-FP. Based on maximum residue levels (MRL) for AMX in pigs established at 50 microg/kg for all tissues, the withdrawal times of AMX in muscle and skin plus fat were estimated (95% tolerance limit and 95% confidence) to fall below the MRL after a withdrawal period of seven days. Levels of AMX in the liver and kidneys were estimated to fall below the MRL after a withdrawal period of four days.     

CORRELATION OF ARTICULAR CARTILAGE THICKNESS MEASUREMENTS MADE WITH MAGNETIC RESONANCE IMAGING,MAGNETIC RESONANCE ARTHROGRAPHY,AND COMPUTED TOMOGRAPHIC ARTHROGRAPHY WITH GROSS ARTICULAR CARTILAGE THICKNESS IN THE EQUINE METACARPOPHALANGEAL JOINT

Osteoarthritis of the metacarpophalangeal joint is common cause of lameness in equine athletes, and is hallmarked by articular cartilage damage. An accurate, noninvasive method for measuring cartilage thickness would be beneficial to screen for cartilage injury and allow for prompt initiation of interventional therapy. The objective of this methods comparison study was to compare computed tomographic arthrography (CTA), magnetic resonance imaging (MRI), and magnetic resonance arthrography (MRA) measurements of articular cartilage thickness with gross measurements in the metacarpophalangeal joint of Thoroughbred horses. Fourteen cadaveric, equine thoracic limbs were included. Limbs were excluded from the study if pathology of the metacarpophalangeal articular cartilage was observed with any imaging modality. Articular cartilage thickness was measured in nine regions of the third metacarpal bone and proximal phalanx on sagittal plane MRI sequences. After intra‐articular contrast administration, the measurements were repeated on sagittal plane MRA and sagittal CTA reformations. In an effort to increase cartilage conspicuity, the volume of intra‐articular contrast was increased from 14.5 ml, to maximal distention for the second set of seven limbs. Mean and standard deviation values were calculated, and linear regression analysis was used to determine correlations between gross and imaging measurements of cartilage thickness. This study failed to identify one imaging test that consistently yielded measurements correlating with gross cartilage thickness. Even with the use of intra‐articular contrast, cartilage surfaces were difficult to differentiate in regions where the cartilage surfaces of the proximal phalanx and third metacarpal bone were in close contact with each other.     

Concentration,Activity and Biochemical Characterization of Myeloperoxidase in Fresh and Post‐Thaw Equine Semen and their Implication on Freezability

Myeloperoxidase (MPO) is a pro‐oxidant enzyme associated with decreased motility in thawed equine semen. This study aimed to describe MPO concentration, activity and subunits in raw and thawed semen and to correlate these data with motilities in raw and thawed semen. Semen samples from five stallions were collected four times. Motilities were assessed in raw and thawed semen. MPO assays were performed in raw seminal plasma, raw sperm‐rich pellet and thawed semen. Total and active MPO concentrations were, respectively, assayed by enzyme‐linked immunosorbent assay and specific immunological extraction followed by enzymatic detection. MPO subunits present in semen were characterized by Western blot. Purified active MPO was added in saline solution and freezing extender to control its activity during freezing procedure. Differences between medians were determined using Kruskal–Wallis test, and correlations were determined using Spearman's test for nonparametric data. Active MPO concentration was low in seminal plasma and thawed semen, but high in pellet (p = 0.0058), as the opposite relation was observed for total MPO concentration (p < 0.0001). In seminal plasma and post‐thaw semen, inactive 86‐kDa MPO precursor was mainly observed. Purified MPO activity was decreased in the extender (p = 0.0286). MPO activity in pellet was highly correlated with thawed progressive motility (r = ?0.5576, p = 0.0086). Inactive MPO precursor and unknown low molecular weight inactive MPO precursor subunits explain low MPO activity in semen. Major MPO activity was observed in pellet, and post‐thaw loss of activity is partially explained by MPO inactivation in extender. Thawed semen motility was negatively correlated with MPO activity in pellet, becoming a potential freezability predictor.     

Vertical distribution and migration of fish larvae in the NW Iberian upwelling system during the winter mixing period: implications for cross‐shelf distribution

The vertical distribution and vertical migrations of fish larvae and implications for their cross‐shelf distribution were investigated in the northern limit of the NE Atlantic upwelling region during the late winter mixing period of 2012. The average positive values of the upwelling index for February and March of this year were far from normal, although the average hydrographic conditions during the period of study were of downwelling and the water column was completely mixed. Fish larvae, most in the preflexion stage, were concentrated in the upper layers of the water column and their distribution was depth stratified, both day and night. However, the larval fish community was not structured in the vertical plane and fish larvae did not show significant diel vertical migration (DVM), although five species showed ontogenetic vertical migration. In regions of coastal upwelling and in the absence of DVM, the location of fish larvae in the water column is crucial for their cross‐shelf distribution. Thus, the cross‐shelf distribution of the six most abundant species collected in this study can be explained by the surface onshore flow associated with coastal downwelling, retaining larvae of the coastal spawning species with a relatively shallow distribution in the shelf region and transporting larvae of slope spawning species onto the shelf. The wide vertical distribution shown by larvae of the offshore spawning species could be an adaptation of these species to ensure that some larvae reach the inshore nursery areas.     

Effect of dietary microminerals in early weaning diets on growth,survival, mineral contents and gene expression in gilthead sea bream (Sparus aurata,L) larvae

In previous studies, combined inclusion of Zn, Mn and Se in early weaning diets improved larval growth, but suggested a potential toxicity by one of these elements. The present study aimed to determine the effect of the single inclusion of Zn, Mn, Se or Cu, their combination (Control+) or their absence (Control?) on larval diets. At the end of the trial, survival was significantly (p < .05) lowest in fish fed C+ diet (17.16 ± 7% mean ± SD), followed by that of larvae fed Mn diet (21.91 ± 7%). The highest survival was obtained by Cu diet (35.27 ± 15%), followed by C? diet (34.58 ± 9%). Cu and Se supplementation significantly improved total length and body weight, in comparison with the C? fish. On the contrary, fish fed Mn and C+ showed the lowest growth. Supplementation with Zn or Cu significantly increased CuZnsod, whereas gpx was significantly upregulated in fish fed Se and C+ diets. ARA/EPA level was significantly highest and DHA/EPA lowest in larvae fed the Cu diet in fish fed C+ diet. The results pointed out the importance of supplementation with Cu, as well as Se and Zn, on early weaning diets for gilthead sea bream, and the potential toxic effect of Mn.     

Increase in auxiliary photoprotective photosynthetic pigments in wheat seedlings induced by Azospirillum brasilense

Inoculation of wheat seedlings with the plant growth-promoting bacterium Azospirillum brasilense Cd was immobilized in alginate microbeads and, without applying any stress, significantly increased the quantity of several photosynthetic pigments, such as chlorophyll a, chlorophyll b, and the auxiliary photoprotective pigments violaxanthin, zeaxanthin, antheroxanthin, lutein, neoxanthin, and β-carotene. This resulted in greener plants with no apparent visible stress. After monitoring the quantity of photosynthetic pigments for 4 weeks, we observed that inoculated plants had higher quantities of pigments in shoot and stem. The greatest difference in the quantity of all pigments between inoculated and noninoculated plants occurred in the first week of growth. Regardless of treatment, the quantity of pigments in stems was three to four times less than the quantity of these pigments in shoots. Application of Azospirillum, either as liquid inoculant or as alginate microbeads, did not alter the positive effect of the bacteria on pigment production or the positive response of the plants towards A. brasilense Cd inoculation.     

Contribution of malolactic fermentation by Oenococcus oeni and Lactobacillus plantarum to the changes in the nonanthocyanin polyphenolic composition of red wine

The changes in the nonanthocyanin phenolic composition during red wine malolactic fermentation carried out spontaneously and by four different starter cultures of the species Oenococcus oeni and Lactobacillus plantarum were examined to determine whether differences in nonanthocyanin polyphenolic compounds could be attributed to the lactic acid bacteria (LAB) strain that performs this important step of the wine-making process. The polyphenolic compounds were analyzed by high-performance liquid chromatography with photodiode array detection and HPLC with electrospray ionization-mass spectrometry detection. The malolactic cultures selected for this study were indigenous wine LAB strains from the A.O.C. Rioja (Spain). Results showed different malolactic behaviors in relation to wine phenolic compositions for O. oeni and L. plantarum, and also, a diversity was found within each group. The hydroxycinnamic acids and their derivatives, the flavonols and their glycosides, the flavanol monomers and oligomers, and trans-resveratrol and its glucoside were the main compounds modified by the different LAB. The wild LAB population exerted a greater impact in the wine content of some of these phenolic compounds than the inoculated selected monocultures of this study.