Identification of viral pathogens in aborted fetuses and stillborn piglets from cases of swine reproductive failure in Spain

The objective of the present study was to determine the presence of recognised abortifacient viruses such as porcine reproductive and respiratory virus (PRRSV), Aujeszky's disease virus (ADV), porcine parvovirus (PPV) and porcine circovirus type 2 (PCV2), in tissues from aborted fetuses and stillborn neonates in cases of late reproductive failure in swine. A total of 293 specimens (fetuses aborted in the last third of gestation and stillborn piglets) from 100 different cases of late-term abortions and premature farrowing from 15 different Spanish provinces were studied. PRRSV was detected in 9/100 cases by RT-PCR. Only 1/100 cases analysed (corresponding to a late-term aborted fetus with a negative PRRSV RT-PCR result) was positive for PCV2 by PCR. Neither ADV (monitored by viral isolation plus antigen detection) nor PPV (monitored by ELISA antigen capture test) infection was identified. The results suggest that PRRSV is one of the most important infectious agents, if not the most relevant one, associated with fetal infection leading to abortion or premature farrowing in Spain. Moreover, other viral pathogens such as ADV, PPV and PCV2 seem to have a minor impact on reproductive disease.     

Pharmacokinetics of azithromycin after i.v. and i.m. administration to sheep

The pharmacokinetics (PK) of azithromycin after i.v. and i.m. injection at a single dosage of 20 mg/kg bodyweight was studied in sheep. Blood samples were collected from the jugular vein until 120 h after dosing for both routes. Plasma concentrations of azithromycin were determined by bioassay. The plasma concentration-time data of azithromycin best fitted a three-compartment model after i.v. administration and a two-compartment model with first-order absorption after i.m. administration. The elimination half-life (t(1/2lambdaz)) was 47.70 +/- 7.49 h after i.v. administration and 61.29 +/- 13.86 h after i.m. administration. Clearance value after i.v. dosing was 0.52 +/- 0.08 L/kg.h. After i.m. administration a peak azithromycin concentration (C(max)) of 1.26 +/- 0.19 mg/L was achieved at 1.24 +/- 0.31 h (t(max)). Area under the curve (AUC) were 38.85 +/- 5.83 mg.h/L and 36.03 +/- 1.52 mg.h/L after i.v. and i.m. administration respectively. Bioavailability obtained after i.m. administration was 94.08 +/- 11.56%. The high tolerability of this i.m. preparation and the favourable PK behaviour such as the long half-life and high bioavailability make azithromycin likely to be effective in sheep.     

Evaluation of the API 50CH and API ZYM systems for rapid characterization of Clavibacter michiganensis subsp. sepedonicus, causal agent of potato ring rot

API 50CH and API ZYM systems were used to characterize fifty-three strains of Clavibacter michiganensis subsp. sepedonicus from different geographic locations and several reference strains of the same and different species, including other potato pathogens. Clavibacter michiganensis subsp. sepedonicus strains displayed a high level of homogeneity, both in carbohydrate utilization and in enzymatic activity. Using API 50CH and API ZYM it was possible to differentiate C. michiganensis subsp. sepedonicus strains from the remaining taxa analysed in this study, which included representative strains of the other subspecies of C. michiganensis as well as other bacterial pathogens affecting potatoes. Therefore, these systems could be used as an effective method to characterize C. michiganensis subsp. sepedonicus. Such a procedure would constitute an alternative system to the conventional nutritional and physiological identification tests currently included in the official methods employed in the European Union to detect and identify this bacterium. The results obtained with the API systems agreed with the current taxonomic classification of C. michiganensis, clearly separating sepedonicus from the other subspecies belonging to this species.     

Residues of diflubenzuron on horse chestnut (Aesculus hippocastanum) leaves and their efficacy against the horse chestnut leafminer, Cameraria ohridella

Residues of the insect growth regulator diflubenzuron were quantified on horse chestnut (Aesculus hippocastanum L.) leaves treated with a diflubenzuron 480 g litre^?1 SC, Dimilin. To analyse the samples, an analytical procedure was developed involving a simple extraction step followed by high‐performance liquid chromatography on an octadecyl‐modified silica column with methanol + 0.01 M ammonium acetate mobile phase. The results showed diflubenzuron to be highly stable on horse chestnut leaves; more than 4 months (127 days) after application, 38% (on average) of the insecticide still remained on/in the leaves. The data confirmed biological observations showing diflubenzuron's long‐term efficacy against the horse chestnut leafminer, Cameraria ohridella Deschka & Dimi?, which is the most important pest of the horse chestnut in Europe. The hypothesis of possible penetration of diflubenzuron into the leaf mass is explored and discussed. Copyright © 2006 Society of Chemical Industry     

Organic amendments affecting sorption, leaching and dissipation of fungicides in soils

Metalaxyl and tricyclazole are two fungicides widely used in Spain in vineyard and rice crops respectively. In this study an investigation has been made of the effect of three organic amendments [two commercial amendments, solid fertiormont (SF) and liquid fertiormont (LF), and a residue from the olive oil production industry, alperujo (OW)] on fungicide fate in soils. Changes in soil porosity on amendment were studied by mercury intrusion porosimetry, sorption-desorption studies were performed by the batch equilibration method, dissipation of metalaxyl and tricyclazole in the soil was studied at - 33 kPa moisture content and 20 degrees C and leaching was studied in hand-packed soil columns. Amendments with SF and LF reduced soil porosity, while OW increased porosity through an increase in pore volume in the highest range studied. Tricyclazole sorbed to soils to a much higher extent than metalaxyl. With some exceptions, sorption of both fungicides increased on amendment, especially in the case of SF-amended soils, which rendered the highest K(oc) values. In soils amended with the liquid amendment LF, sorption either remained unaffected or decreased, and this decrease was much higher in the case of metalaxyl and a soil with 70% clay. In this clay soil, amendment with OW, of very high soluble organic matter content, also decreased metalaxyl sorption. Tricyclazole is more persistent in soil than metalaxyl, and both fungicides were found to be more persistent in amended soils than in unamended soils. Leaching of metalaxyl and tricyclazole in soil columns was inversely related to sorption capacity. The low recoveries of tricyclazole in leachates and in soil columns when compared with metalaxyl, a less persistent fungicide, were attributed to diffusion into micropores and to increase in sorption with residence time in the soil, both processes favoured by the low mobility of tricyclazole.     

Evaluation of metabolic detoxifying enzyme activities and insecticide resistance in Frankliniella occidentalis

The western flower thrips Frankliniella occidentalis (Pergande) is a very significant pest of a number of different agricultural crops in the south-east of Spain. The importance of thrips as a pest is not due mainly to the direct damage inflicted on the plant, but to the loss in commercial value which occurs as a consequence of the development of dark spots caused by the tomato spotted wilt virus (TSWV) which they transmit. The economic threshold is therefore almost zero, which enhances the problems of resistance management. The present work is part of a global project that attempts to evaluate the status of insecticide resistance in field populations of thrips obtained from several agricultural crops. We have studied, in either individual or pooled insects, some enzyme systems classically related to detoxification of insecticides: esterase and glutathione-S-transferase (GST). The activity of these enzymes from laboratory populations selected with various classes of insecticides has also been measured using several appropriate substrates. An increase in GST mean activity was found in two field-collected strains. Differences in frequency distributions of esterase and GST activities were found for both field-collected strains and for a laboratory strain selected with acrinathrin. These activities were compared with those of a wild-type reference strain.     

Relationships between agronomic factors and epidemics of Phytophthora branch canker of citrus in southwestern Spain

Phytophthora branch canker, caused by Phytophthora citrophthora, has been an increasing problem in clementine (Citrus reticulata) production in Spain during last years. The disease was particularly severe in the new citrus-growing areas of the southwestern coastal areas in Huelva Province. Recent studies revealed that disease emergence was not related to either genetic drift or host specificity changes in P. citrophthora population. Therefore, the possible association of agronomic factors with the disease was investigated. A total of 110 orchards were selected arbitrarily from the main citrus-growing areas in Huelva Province. The presence of branch cankers together with agronomic factors including soils, cultivars, rootstocks, irrigation, pruning, techniques to improve fruit production, fungicide treatments, presence of brown rot of fruit and frost damage were recorded. Logistic regression analysis was used to detect correlations between the agronomic factors studied and disease prevalence. Phytophthora branch canker was significantly associated with mature clementine orchards. Sweet orange and hybrid cultivars as well as young clementine orchards were less affected by the disease. Although disease was less frequent in Salorthid soils, alternative high resolution procedures are required to draw conclusions about the effect of soil properties on disease prevalence. As in other Phytophthora-induced diseases, soil flooding during the rainy season was correlated positively with the prevalence of branch cankers. Improving fruit production by branch scoring showed a strong positive correlation with Phytophthora branch canker. This is the first time that girdling has been associated with Phytophthora disease epidemics on a fruit tree crop, but further research is needed to determine the cause of this relationship. Cultural practices including pruning, regulated deficit irrigation, additional phosphonate sprays, and abiotic and disease factors such as frost damage and presence of brown rot of fruit were not significantly correlated with disease prevalence.     

Dispersal of conidia of Fusicladium eriobotryae and spatial patterns of scab in loquat orchards in Spain

Dispersal of conidia of Fusicladium eriobotryae, the causal agent of loquat scab, was investigated in two loquat orchards in Spain from 2010 to 2012. A volumetric spore sampler, horizontally and vertically orientated microscope slides, and rain collectors were placed in loquat fields to trap conidia of F. eriobotryae. No conidia were collected in the volumetric sampler, and highly variable numbers of conidia were collected in the rain gatherers. Large numbers of conidia were collected by microscope slides, particularly by those held in a horizontal orientation compared with those held in a vertical orientation. Approximately 90 % of the F. eriobotryae conidia were collected during rainy periods. Based on ROC and Bayesian analysis, using ≥ 0.2 mm rainfall as a cut-off value resulted in a high probability of correctly predicting actual conidial dispersal, and had a low probability of failing to predict actual conidial dispersal. Based on the index of dispersion and the binary power law, the incidence of loquat scab on fruit was highly aggregated in space between and within trees, and aggregation was influenced by disease incidence. Our results demonstrate, for the first time, that F. eriobotryae is dispersed mainly in rain splash. The results will be integrated into a mechanistic, weather-driven, disease prediction model that should help growers to minimize fungicide application for the management of loquat scab.