Design of Division Specific Primers of Ralstonia solanacearum and Application to the Identification of European Isolates

A PCR diagnostic test for detection of Ralstonia solanacearum at the infraspecific level was developed, based on polymorphisms within the 16S rRNA gene sequence. Partial sequences of this gene were determined for three French isolates which showed the characteristics of R. solanacearum subdivision 2a described by Taghavi et al. (1996). Oligonucleotide primers were designed to incorporate the nucleotide triplet (at positions 458–460 of the 16S rRNA gene) which differs between divisions 1 and 2 16S rRNA sequences of R. solanacearum isolates. A simple PCR test unambiguously revealed the division of each isolate. The PCR test was useful for identification of isolates of R. solanacearum from Europe.     

Initial distance to Kalmia angustifolia as a predictor of planted conifer growth

New Forests - In the Canadian boreal forest, conifer plantations are mainly used to overcome poor natural regeneration following harvesting or wildfires. However, competitive interactions can...     

Relationship between aggressiveness of <Emphasis Type="Italic">Stagonospora </Emphasis>sp. isolates on field and hedge bindweeds,and <Emphasis Type="Italic">in vitro</Emphasis> production of fungal metabolites cercosporin,elsinochrome A and leptosphaerodione

Stagonospora convolvuli LA39, an effective biocontrol agent of Convolvulus arvensis (field bindweed) and Calystegia sepium (hedge bindweed) produces phytotoxic metabolites leptosphaerodione and elsinochrome A. Stagonospora isolate 214Caa produces the toxin cercosporin. If toxic metabolite production is not linked to the pathogenic ability of the fungus on bindweeds, selection of aggressive strains with limited or no production of the metabolites would reduce any perceived risk of using strains of the fungus as a mycoherbicide. Therefore, 30 isolates of Stagonospora sp. including LA39 and 214Caa were characterised for aggressiveness on both bindweeds, and production of the three metabolites. Nine isolates were more aggressive than LA39 on both bindweeds. Classification of isolates based on metabolite type agreed largely with previous similar characterisation based on polymerase chain reaction-restriction fragment length polymorphism of internal transcribed spacer of ribosomal DNA. Cercosporin producers produced neither leptosphaerodione nor elsinochrome A and together with isolates that produce none of the three metabolites, were less pathogenic on bindweeds. Conversely, there was a positive correlation between elsinochrome A and leptosphaerodione production, and each was positively correlated with aggressiveness of isolates on both bindweeds. Generally, any isolate where elsinochrome A was not detected was not aggressive on any of the two bindweeds. This probably implies that selecting elsinochrome A-negative, but aggressive Stagonospora strain(s) may be difficult. However, aggressive isolates may not produce elsinochrome A in planta at levels that could constitute any risk in the environment. In a preliminary attempt to determine the levels of elsinochrome A and leptosphaerodione produced in diseased bindweeds, none of the toxins was detected in Stagonospora infected bindweed leaves. Detailed investigation focusing on the detection and quantification of in planta production of elsinochrome A by Stagonospora isolates, and determination of the fate of elsinochrome A in the environment, and its relationship with leptosphaerodione may be essential. Similarly, development of molecular tools to monitor the mycoherbicide following field application is vital.     

Evaluation of experimental methods to induce congenital hypothyroidism in guinea pigs for use in the study of congenital hypothyroidism in horses

OBJECTIVE: To develop a method to reliably induce congenital hypothyroidism in guinea pigs (Cavia porcellus) and assess similarities between the resultant developmental abnormalities and those described in horses with congenital hypothyroidism. ANIMALS: 35 female guinea pigs and their offspring. PROCEDURE: Guinea pigs were allocated to control groups or groups treated with a low-iodine diet before and throughout gestation; an s.c. injection of 100 or 200 microCi of radioactive iodine 131 (131I) on day 40 of gestation; or 0.1% propylthiouracil (PTU) continuously in the drinking water, beginning day 3 or 40 of gestation. In all groups, assessments included gestation duration, litter size, proportion of stillborn pups, and laboratory analyses in live pups and dams; postmortem examinations were performed on all pups and dams and selected tissues were examined histologically. RESULTS: Compared with control animals, pups from dams receiving a low-iodine diet or 131I s.c. had mild changes in their thyroid glands but no grossly or radiographically detectable lesions of hypothyroidism. Pups from dams receiving PTU were often stillborn (24/27 pups) and had enlarged thyroid glands (characterized by large, variably sized follicles of tall columnar epithelium and little or no colloid), an incomplete coat, and radiographically detectable skeletal dysgenesis. CONCLUSIONS AND CLINICAL RELEVANCE: Many of the lesions detected in guinea pig pups from the experimentally treated dams were similar to those described in foals with congenital hypothyroidism. Experimental induction of congenital hypothyroidism in guinea pigs may be useful for the study of naturally occurring congenital hypothyroidism in horses.     

Effect of subacute oral doses of nivalenol on immune and metabolic defence systems in mice

Nivalenol (NIV) is a toxic Fusarium secondary trichothecene metabolite occurring naturally in cereal grains. In order to evaluate the no observed adverse effect level (NOAEL), we tested the effects of a large array of oral doses of this toxin for responses on plasma biochemistry, the immune system and hepatic drug metabolism in mice. C57Bl6 mice received oral doses of toxin (0.014, 0.071, 0.355, 1.774 or 8.87 mg/kg bw) 3 days a week for 4 weeks. Only the highest dose of NIV induced an increase in plasma phosphate, decreases in plasma urea and immunoglobulin M and additional changes like increases in plasma alkaline phosphatase and immunoglobulin G. Interleukin 4 production was increased in cultured murine splenocytes. Regarding liver drug metabolising enzymes, the only glutathione transferase activity accepting 1-chloro-2,4-dinitro-benzene as substrate was transiently increased in mice receiving low doses (0.071 and 0.355 mg/kg bw) of NIV. Regarding the cytochrome P450 monooxygenases, no significant change was observed in ethoxyresorufin O-deethylase activity whereas both methoxyresorufin and pentoxyresorufin O-dealkylase activities were decreased by 38-45% for the highest dose (8.87 mg/kg bw) of NIV. However, when analysed by Western blot analysis, the protein expression of mouse P450 1a, 2b, 2c, 3a and 4a subfamilies was unchanged in animals receiving NIV. In conclusion, the NOAEL of this toxin in our study was 1.774 mg/kg bw, corresponding to an exposure to 5 ppm contaminated food. Indeed hepatotoxicity appears in the only mice treated with a five fold higher oral dose of 8.87 mg/kg bw of NIV. Such exposure levels appear to be by far higher than the maximal natural occurrence measured in European cereals, known to range from 0.34 to 1.86 ppm.