Evaluation of diagnostic and quantitative PCR for the identification and severity assessment of eyespot and sharp eyespot in winter wheat

Diagnostic and quantitative polymerase chain reaction (PCR) provided clarification of the causes of symptoms and the extent of infection by eyespot ( Tapesia spp.) and sharp eyespot ( Rhizoctonia cerealis ) on winter wheat at early growth stages. Disease assessments made before stem extension, when decisions to apply fungicides are usually made, often did not agree with the pathogen diagnoses using PCR, suggesting that such early visual diagnoses may be unreliable. Visual and PCR diagnoses made on stems in summer generally supported each other, but there were often discrepancies in relating disease severity to amounts of pathogen present when determined by regression analyses of incidence or severity of symptoms on amount of pathogen DNA. Mixed symptoms caused by different pathogens may sometimes have been confounded. Relationships between symptoms and DNA of eyespot pathogens were less clear on some cultivars, often those with least disease. Sharp eyespot symptoms had a stronger relationship to DNA of its pathogen. Significant regressions often accounted for a small percentage of the variance, suggesting either that pathogens not assayed were contributing to symptoms or that lesions were in some cases persisting longer into the season than pathogen DNA. The frequency of pathogen detection before stem extension was a poor predictor of the amounts of pathogen DNA measured later in the season.     

用RAPD分析和DNA指纹鉴别玉米小斑病菌O、C、T三个小种

 目前通过病理学鉴定已发现玉米小斑病菌存在分别对玉米雄性不育T型、C型细胞质专化侵染的T小种和C小种(无细胞质专化致病性的相应称为O小种)。     

Determination of deoxynivalenol and nivalenol chemotypes of Fusarium culmorum isolates from England and Wales by PCR assay

Functioning Tri13 and Tri7 genes are required for the production of nivalenol and 4-acetyl nivalenol, respectively, in Fusarium species producing type B trichothecenes. Mutations have been identified in isolates which are able to produce deoxynivalenol (DON) but unable to convert this to nivalenol (NIV). In such isolates of Fusarium culmorum , the Tri7 gene is deleted entirely. PCR assays specific for functional and nonfunctional/deleted versions of Tri7 and Tri13 were used to determine the ability of 153 single spore isolates of F. culmorum to produce the 8-ketotrichothecenes deoxynivalenol and nivalenol. The isolates were collected from 76 different locations across England and Wales between 1994 and 2002. Four isolates were also obtained from one field in Scotland. Both DON and NIV chemotypes of F. culmorum were identified, with DON chemotypes predominating overall. In addition, all DON chemotypes were shown to produce 3-acetyl DON using primer sets developed to Tri3 . From fields where more than one F. culmorum isolate was obtained, isolates were not exclusively of a single chemotype. Differences in the distribution of DON and NIV chemotypes were identified, with a greater proportion of NIV chemotypes present in the south and west of England and Wales, whereas a greater proportion of DON chemotypes were found in the north and east of England. Seasonal differences in the ratio of DON:NIV chemotypes were indicated. However, these were related to seasonal variation in the distribution of F. culmorum .     

Acute cortisol responses of calves to four methods of dehorning by amputation

Objective To measure the plasma cortisol response in calves dehorned by four different methods (scoop, guillotine shears, saw, embryotomy wire) for 9 h after dehorning.
Design A physiological study with controls.
Procedure Horn amputation was carried out on calves restrained manually in a race.
Results The four methods of dehorning provoked similarly increased cortisol responses which lasted for 6 h. During the first hour after dehorning the plasma cortisol concentrations were similar to those following ACTH injection. The overall cortisol response to control handling was about 30% of the responses to dehorning.
Conclusions The similarity of the cortisol responses produced by the four methods of dehorning suggests that the distress experienced by calves following dehorning by amputation is similar regardless of method used.     

Suppression of ovarian oxytocin secretion after intra-luteal administration of the arachidonate 5-lipoxygenase inhibitor BWA4C in the ewe

The effect of an intra-luteal injection of the 5-lipoxygenase (5-LO) inhibitor BWA4C (2 mg in 50 μ1 dmso) on the secretion of oxytocin (OT) from the corpus luteum in response to a close-arterial infusion of prostaglandin (PG)-F_2α (5 ng mint) was examined in anaesthetised sheep. Within 30 minutes of administration both basal (pre-infusion) and PGF_2α stimulated OT release into the posterior vena cava were significantly (P<0·01) reduced. These results are consistent with the proposition that 5-LO products of arachidonic acid may modulate OT secretion from the ovine corpus luteum.     

Partial characterization of the extracellular matrix released onto hydrophobic surfaces by conidia and conidial germlings ofColletotrichum graminicola

The extracellular matrix (ECM) released by conidia ofColletotrichum graminicolawhen incubated on artificial hydrophobic substrata was investigated. Time-course experiments revealed that proteins and carbohydrates were released in the ECM of both ungerminated and germinated conidia. Stains for protein and carbohydrate revealed that part of the ECM remained attached to the substratum after the fungus had been removed. The ECM isolated from substrata where ungerminated conidia had adhered and from substrata where germinated conidia had adhered exhibited the same profiles when analysed by SDS-PAGE for proteins and glycoproteins and by HPLC for carbohydrate analysis. The ECM was found to be composed of glycoproteins with a molecular weight of around 200 kDa. Deglycosylation of the isolated ECM material resulted in the release of a protein with a molecular weight of approx. 116 kDa, indicating that the original protein was extensively glycosylated. Carbohydrate analysis demonstrated that mannose was the predominant sugar. That the ECM remained attached to the surface of the hydrophobic substrata suggests that it has a role in adhesion.     

Refinement of PCR-detection of Fusarium avenaceum and evidence from DNA marker studies for phenetic relatedness to Fusarium tricinctum

Existing PCR-based assays for the detection of the cereal pathogen Fusarium avenaceum were found to cross-react with F. tricinctum . An investigation into the phenetic relationship between these two species using two different marker systems revealed a close relationship between them despite their being from separate taxonomic sections. Whilst RFLP differences in the ITS regions surrounding the nuclear 5.8 S rDNA were insufficient to discriminate between isolates of the two species, they were clearly differentiated by RAPD profiling. RAPD fragments from F. avenaceum isolates were screened for hybridization to F. tricinctum DNA on Southern blots. One of 12 selected RAPD fragments showed no hybridization to genomic DNA from F. tricinctum . This fragment was cloned and sequenced, and the sequence obtained was used to design PCR primers. The primers were found to be specific for F. avenaceum, with no cross-reactions obtained with F. tricinctum or any other wheat pathogen assayed. The primers were able to differentiate between the two species in infected plant material, in contrast to the earlier assays.     

Predominance and association of pathogenic fungi causing Fusarium ear blightin wheat in four European countries

Two years of field sampling aimed to establish the predominance and association among the fungal pathogens causing Fusarium ear blight (FEB) in four European countries (Hungary, Ireland, Italy and the UK). A PCR-based method was used to detect four Fusarium species and two varieties of Microdochium nivale present in the samples. The prevalence of FEB pathogens differed significantly between countries. Overall, all pathogens were commonly detected in Ireland and to a lesser extent in the UK. In contrast, only two species, F. graminearum and F. poae, were regularly detected in Italy and Hungary. Fusarium culmorum was rarely detected except in Ireland. Log-linear models were used to determine whether there is the independence of the six FEB pathogens at each sampling site. Significant two-pathogen interactions were frequently observed, particularly in harvest samples; all these significant two-pathogen interactions were of the synergistic type, except between F. poae and F. culmorum, and were generally consistent over the 2 years and four countries. Fusarium graminearum and F. poae were least frequently involved in two pathogen interactions but were involved in most of the nine significant three-pathogen interactions. However, only the interaction between F. graminearum, F. avenaceum and F. poae was significant in both years. Potential implications of the present results in FEB management are discussed.     

Use of continuous positive airway pressure in the acute management of laryngeal paralysis in a cat

Continuous positive airway pressure (CPAP) has been is used widely in humans to manage obstructive sleep apnoea syndrome, but it has not been widely used in animals. A brachycephalic cat, with previously undiagnosed laryngeal paralysis, that developed acute upper respiratory tract obstruction on recovery from anaesthesia, is presented. The condition was managed by CPAP, delivered via a facial mask.