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.     

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.     

Piquerol a and diacetyl piquerol act as energy transfer inhibitors of photosynthesis

ATP synthesis and phosphorylating (coupled) electron flow of freshly lysed pea chloroplasts were inhibited by piquerol A and diacetyl piquerol, the latter being more active. H + -uptake, basal and uncoupled electron transport were not affected by either compound, indicating that they act as inhibitors of photophosphorylation at the H + -ATPase complex.     

Modelling the microbial breakdown of pesticides in soil using a parameter estimation technique

Results of field measurements and laboratory experiments were used to simulate the behaviour of 1,3-dichloropropene (1,3-D) in a loamy sand soil. Microbial activity was described using pre-set conditions to compute transformation rates as dependent on compound concentrations and temperature. These kinetics could only be analysed using non-linear transformation rates. To link the development of microbial populations and the consumption of the compound over time, an iterative technique was used to estimate the necessary parameters.     

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.     

Study of Resistance to Orobanche ramosa in Host (Oilseed Rape and Carrot) and Non-host (Maize) Plants

Orobanche ramosa is a parasitic Angiosperm responsible for severe yield losses in several economical crops. It is a serious threat in oilseed rape in France and Morocco and is appearing in carrot crops in Morocco. In this study, several varieties of oilseed rape and carrot were screened in order to identify resistant cultivars and to characterize the resistance mechanisms involved. All the 15 oilseed rape varieties tested were susceptible. In carrot, the varieties 'Colmar à coeur rouge' and 'Nantaise demi-longue' were susceptible, whereas 'Palaiseau' and 'Buror' were resistant. In the susceptible 'Colmar à coeur rouge' carrot no defence reactions were found and the development of the parasite inhibited carrot tap root formation. In the resistant carrot varieties, the parasite germinated, became attached to the host root but became necrotic before emergence. In 'Buror' carrot, formation of a mechanical barrier was associated with the restriction to the cortex of the parasite. In maize cv. 'Vigni', a non-host of O. ramosa, thickening of xylem vessels, cell divisions in the central cylinder and formation of an encapsulation layer were observed in association with restricted development of Orobanche haustoria.     

Isolation of viral haemorrhagic septicaemia virus from mummichog, stickleback, striped bass and brown trout in eastern Canada

Viral haemorrhagic septicaemia virus (VHSV) was isolated from mortalities occurring in populations of mummichog, Fundulus heteroclitus, stickleback, Gasterosteus aculeatus aculeatus, brown trout, Salmo trutta, and striped bass, Morone saxatilis, in New Brunswick and Nova Scotia, Canada. The isolated viral strains produced a cytopathic effect on the epithelioma papillosum cyprini cell line. Serum neutralization indicated the virus was VHSV and sequencing identified the rhabdovirus isolates as the North American strain of VHSV. Phylogenetic analysis indicated that the isolates are closely related and form a distinguishable subgroup of North American type VHSV. To our knowledge, this is the first report of VHSV in mummichog and striped bass.     

Evaluation of biochemical analytes in vitreous humor collected after death in West Indian manatees

OBJECTIVE: To evaluate biochemical analytes in vitreous humor collected after death in West Indian manatees (Trichechus manatus latirostris). DESIGN: Prospective study. ANIMALS: 73 West Indian manatees of various ages and both sexes. PROCEDURE: The condition of manatee carcasses submitted for routine postmortem examination was categorized as fresh, moderately autolyzed, or severely autolyzed. The eyes were removed, frozen for shipping, and thawed on arrival at the laboratory. Vitreous humor was extracted, and various biochemical analytes were measured. Values for each analyte were compared with published reference ranges for corresponding biochemical analytes in serum. Values were also compared among the carcass condition groups. RESULTS: Creatinine and urea nitrogen concentrations in vitreous humor were significantly higher in severely autolyzed carcasses than in fresh carcasses. Potassium concentrations in vitreous humor were significantly higher in moderately autolyzed carcasses than in fresh carcasses but were highly variable in severely autolyzed carcasses. These data were consistent with those reported in other species. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that measurement of biochemical analytes in vitreous humor of manatees is feasible and has potential for assisting in the postmortem diagnosis of certain metabolic, renal, and nutritional diseases; determining severity of autolysis at time of necropsy; and determining the time of occurrence of human-induced trauma.     

Improved diagnosis of powdery scab (Spongospora subterranea f.sp.subterranea) symptoms on potato tubers (Solanum tuberosum L.)

Summary Based on visual inspection, discrimination between common scab (Streptomyces spp.) and powdery scab (Spongospora subterranea) can be difficult. Inspections are performed on unwashed samples, incidentally supported by microscopic examination. During 1994–1996 surveys were performed in The Netherlands on tubers with symptoms resembling common scab. Under microscopic assessment nearly all samples showed the presence of structures resembling cystosori (sporeballs) ofS. subterranea. At that time confirmation using alternative techniques was not possible. In 2003 research was undertaken to clarify the situation with respect to scab on potato tubers in The Netherlands. One hundred and eighteen scab samples were extensively tested forS. subterranea. All samples were digitally photographed, microscopically examinated and tested with real-time PCR and DAS-ELISA. Use of these modern methods resulted in a clear picture of symptoms that can be attributed toS. subterranea. A lot of scab samples with structures resembling cystosori could not be confirmed as contaminated withS. subterranea.