Detection and quantification of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis> in irrigation waters

A quantitative RT real-time PCR method was developed for the detection and quantification of Tomato mosaic virus (ToMV) in irrigation waters. These have rarely been monitored for the presence of plant pathogenic viruses, mostly due to the lack of efficient and sensitive detection methods. The newly developed method presented here offers a novel approach in monitoring the health status of environmental waters. ToMV was reliably detected at as low as 12 viral particles per real-time PCR reaction, which corresponds to the initial concentration of approximately 4.2 × 10^?10 mg (6,300 viral particles) of ToMV per ml of sample. The sensitivity of the method was further improved by including the Convective Interaction Media^® (CIM) monolithic chromatographic columns for quick and efficient concentration of original water samples. Seven out of nine water sources from different locations in Slovenia tested positive for ToMV, after concentrating the sample. Four samples tested ToMV-positive without the concentrating procedure. The presence and integrity of infective ToMV particles in the original sample, as well as in the chromatographic fraction, was confirmed using different methods from test plants, DAS ELISA to electron microscopy and real-time PCR. In this study, we propose a unique and simple diagnostic scheme for rapid, efficient, and sensitive monitoring of irrigation waters that could also be adopted for other plant, human or animal viruses.     

The corky root rot pathogen Pyrenochaeta lycopersici secretes a proteinaceous inducer of cell death affecting host plants differentially

Pathogenic isolates of Pyrenochaeta lycopersici, the causal agent of corky root rot of tomato, secrete cell death in tomato 1 (CDiT1), a homodimeric protein of 35 kDa inducing cell death after infiltration into the leaf apoplast of tomato. CDiT1 was purified by fast protein liquid chromatography, characterized by mass spectrometry and cDNA cloning. Its activity was confirmed after infiltration of an affinity-purified recombinant fusion of the protein with a C-terminal polyhistidine tag. CDiT1 is highly expressed during tomato root infection compared with axenic culture, and has a putative ortholog in other pathogenic Pleosporales species producing proteinaceous toxins that contribute to virulence. Infiltration of CDiT1 into leaves of other plants susceptible to P. lycopersici revealed that the protein affects them differentially. All varieties of cultivated tomato (Solanum lycopersicum) tested were more sensitive to CDiT1 than those of currant tomato (S. pimpinellifolium). Root infection assays showed that varieties of currant tomato are also significantly less prone to intracellular colonization of their root cells by hyphae of P. lycopersici than varieties of cultivated tomato. Therefore, secretion of this novel type of inducer of cell death during penetration of the fungus inside root cells might favor infection of host species that are highly sensitive to this molecule.     

Parameters of inner quality of the apple scab resistant and susceptible apple cultivars (Malus domestica Borkh.)

Individual organic acids and sugars were analysed in the fruits of scab resistant and susceptible apple cultivars. The total sugars ranged between 128.2 and 191.6 g/kg, and the total organic acid between 5.1 and 13.4 g/kg. In the flesh and peels of different apple varieties single phenolics (gallic, protocatehuic, chlorogenic, caffeic, ferulic and p-coumaric acid, phloridzin, epicatechin, catechin, quercitrin and rutin) were analysed together with their total phenolic content (TPC). ‘Golden Delicious’ was the cultivar with the lowest TPC whereas ‘Rubinola’, ‘Jonagold’ and ‘Goldrush’ had the highest level of TPC in the pulp. Peels showed a 2–9 times higher phenolic content than the pulp. ‘Goldrush’ had the highest content of TPC in its peel. The total antioxidant capacity of peels was about 2–5 times higher than respective pulps. Scab resistant apple cultivars had significantly higher content of some single and total phenolic contents in comparison with the scab susceptible, especially the pulp.     

IMAGING DIAGNOSIS—CONVENTIONAL AND FUNCTIONAL MAGNETIC RESONANCE IMAGING OF A BRAIN ABSCESS IN A GOAT

A 2‐month‐old female goat was presented for depressed mental status and multifocal central neurologic signs 3 weeks after hot‐iron disbudding. Conventional magnetic resonance imaging (MRI) findings included a large intra axial mass in the left frontal lobe that was T2 hyperintense and T1 hypointense centrally with a contrast‐enhancing peripheral capsule and perilesional T2 hyperintensity. A restrictive pattern was present in diffusion‐weighted imaging. Magnetic resonance spectroscopy demonstrated an increased amount of succinate, acetate, amino acids, lipids; minimal amounts of lactate; and decreased amounts of N‐acetyl aspartate and choline. A cerebral abscess due to Trueperella pyogenes was confirmed from necropsy and tissue culture.     

Pharmacokinetics of carprofen in anaesthetized pigs: a preliminary study

ObjectiveTo investigate the pharmacokinetics of carprofen after a single intravenous (IV) dose and multiple oral doses administered to pigs undergoing electroporation of the pancreas.Study designProspective experimental study.AnimalsA group of eight female pigs weighing 31.74 ± 2.24 kg (mean ± standard deviation).MethodsCarprofen 4 mg kg^?1 was administered IV after placement of a central venous catheter during general anaesthesia with isoflurane. Blood samples were collected 30 seconds before and 5, 10, 20, 30 and 60 minutes and 2, 4, 6, 8, 12 and 24 hours after carprofen administration. Subsequently, the same dose of carprofen was administered orally, daily, for 6 consecutive days and blood collected at 36, 48, 60, 72, 96, 120, 144 and 168 hours after initial carprofen administration. Plasma was analysed using liquid chromatography with mass spectrometry. Standard pharmacokinetic parameters were calculated by compartmental analysis of plasma concentration–time curves. Data are presented as mean ± standard error.ResultsThe initial plasma concentration of IV carprofen was estimated at 54.57 ± 3.92 μg mL^?1 and decreased to 8.26 ± 1.07 μg mL^?1 24 hours later. The plasma elimination curve showed a bi-exponential decline: a rapid distribution phase with a distribution half-life of 0.21 ± 0.03 hours and a slower elimination phase with an elimination half-life of 17.31 ± 3.78 hours. The calculated pharmacokinetic parameters were as follows: the area under the plasma concentration–time curve was 357.3 ± 16.73 μg mL^?1 hour, volume of distribution was 0.28 ± 0.07 L kg^?1 and plasma clearance rate was 0.19 ± 0.009 mL minute^?1 kg^?1. The plasma concentration of carprofen, administered orally from days 2 to 7, varied from 9.03 ± 1.87 to 11.49 ± 2.15 μg mL^?1.Conclusions and clinical relevanceCarprofen can be regarded as a long-acting non-steroidal anti-inflammatory drug in pigs.