Evidence for the Predisposition of Fungicide-Resistant Isolates of Venturia inaequalis to a Preferential Selection for Resistance to Other Fungicides

In the United States, populations of the apple scab pathogen Venturia inaequalis have progressed through three consecutive rounds of fungicide resistance development, first to dodine, then to the benzimidazoles, and most recently to the sterol demethylation inhibitors (DMIs). Analysis of extensive monitoring data have to date provided no indication of detectable cross-resistance or partial cross-resistance of V. inaequalis populations to the three unrelated classes of fungicides prior to the selection of resistant subpopulations. However, in this study, resistance to both benomyl and DMIs developed to significantly higher frequencies within the previously established dodine-resistant population than in the population sensitive to dodine. Accelerated selection of phenotypes double resistant to dodine and the DMI fenarimol was apparent over the course of distinct seasons of apple scab management with either dodine or fenarimol. The data provide evidence for an accelerated speed of resistance development among phenotypes of V. inaequalis already resistant to an unrelated fungicide. This finding represents a departure from the previous model, which assumed entirely independent rounds of resistance developments. The data indicate that phenotypes of V. inaequalis might not only be selected for the trait of fungicide resistance but also for traits allowing a more flexible response to changes in the environment where they compete.     

The life cycle of Dichelyne (Cucullanellus) minutus (Nematoda: Cucullanidae)

Mature specimens of the nematode Dichelyne (Cucullanellus) minutus (Rudolphi, 1819) (Ascaridida, Cucullanidae, Seuratoidea) were obtained from the intestine of flounder Platichthys flesus (L.) caught in the Oresund, Denmark. Plaice Pleuronectes platessa L. and common goby Pomatoschistus microps (Kr?yer) also harbour this species. The eggs embryonate on the seabottom. Larvae about 440 microm long, and believed to be in their third stage, hatch from the eggs. These larvae are not directly infective to flounders or gobies. The polychaete Nereis diversicolor O.F. Muller acts as obligatory intermediate host. Experimental infections showed that larvae >600 microm long and provided with achitinous tooth survived in flounder and common goby. The third-stage larvae moult to fourth-stage larvae in the fish gut wall. Mature worms occur in the lumen of the anterior part of the intestine. All developmental stages may be transferred from one flounder to another; thus the flounder may acquire the parasite also by devouring infected gobies.     

The Use of a GUS Transformant of Trichoderma Harzianum, Strain T3a, to Study Metabolic Activity in the Spermosphere and Rhizosphere Related to Biocontrol of Pythium Damping-off And Root Rot

The activity of Trichoderma harzianum in the spermosphere and rhizosphere of different plant species was studied by use of a beta-glucuronidase (GUS) transformant (strain T3a). Hereby, direct observation of micro-habitats supporting metabolic activity of T. harzianum is reported. Germination of conidia and mycelial growth were not supported by exudates from healthy roots of various ages. Instead, growth and activity of T. harzianum depended on access to dead organic substrates such as seed coats, decaying roots, and wounds, including those caused by infecting pathogens. A correlation between the GUS activity of T. harzianum and the biomass of Pythium ultimum in infected roots was established. On the basis of our observations, we suggest that the biocontrol ability of T. harzianum involves competition with the pathogen for substrates including the seed coat, and wounded or infected root tissue.     

Detection of the German grapevine yellows (Vergilbungskrankheit) MLO in grapevine,alternative hosts and a vector by a specific PCR procedure

A polymerase chain reaction procedure was developed which enables specific amplification of a ribosomal sequence from the mycoplasmalike organism (MLO) associated with German grapevine yellows (Vergilbungskrankheit, VK) and stolbur-related diseases of solanaceous plants. Successful amplification from all samples prepared from various cultivars collected in different viticultural areas indicates that the causal agent is a relatively homogeneous organism. Amplification was also achieved with template DNA prepared from naturally infected weeds in vineyards such asConvolvolus arvensis andSolanum nigrum, and from the planthopperHyalesthes obsoletus that was collected in the vineyards. Feeding of insects of this species on grapevine seedlings resulted in the development of typical yellows symptoms by the grapes.H. obsoletus could therefore be identified as a vector of Vergilbungskrankheit.Abbreviations FD Flavescence dorée - GY Grapevine yellows - MLO Mycoplasmalike organism - PCR Polymerase chain reaction - RFLP restriction fragment length polymorphism - VK Vergilbungskrankheit (German grapevine yellows)     

A new isolate of the nematophagous fungus Duddingtonia flagrans a biological control agent against free-living larvae of horse strongyles

An experiment was carried out in 1997 to test the efficacy of an isolate of the microfungus Duddingtonia flagrans against free-living stages of horse strongyles under conditions in the field and to assess the eventual effect of the fungus on the normal degradation of faeces. Faecal pats were made from faeces of a naturally strongyle infected horse, which had been fed fungal material at a dose level of 106 fungal unit/kg bwt. Control pats without fungi were made from faeces collected from the same animal just before being fed fungi. Faecal cultures set up for both groups of faeces to monitor the activity of the fungus under laboratory conditions showed that the fungus significantly reduced the number of infective third-stage larvae (L3) by an average of 98.4%. Five faecal pats from each batch of faeces were deposited on pasture plots at 3 times during spring-summer. The herbage around each pat was sampled fortnightly to recover L3 transmitted from faeces. The results showed that the herbage infectivity around fungus-treated pats was reduced by 85.8-99.4%. The remaining faecal material at the end of each sampling period was collected, and the surviving L3 were extracted. Significantly fewer larvae were recovered from the fungus-treated pats. Analysis of wet and dry weight of the collected pats, as well as their organic matter content, were performed to compare the degradation of faeces of both groups. The results indicated that the presence of the fungus did not alter the degradation of the faeces.     

Experimental investigations into the effect of Escherichia coli and Clostridium perfringens on the steroid estrone

Together with the faecal and the urinary discharge of animals and humans natural and synthetic estrogenes are excreted into the environment. About their degradation in the environment only few facts are known. Their reduction is probably of microbial enzymatical nature. For looking into that question it was investigated by in-vitro-attempts whether Escherichia coli or Clostridium perfringens influence the estrone. Therefore estrone was incubated together with the microorganisms tested (experimental groups) and without those (control groups) in nutrient medium for these bacteria over 48 hours with 37 degrees C (aerobic) or 42 degrees C (anaerobic). For the executed procedure an enzyme immunoassay was used. It could be seen that E. coli does not show a provable effect on the estrone concentration under this conditions. Whereas C. perfringens might have a potency for the degradation of the hormone tested. The experiments shows that there exists obviously a difference between estrone and 4-pregnene-20 beta-o1-3-one in the same system of test.     

Spontaneous vaginal rupture in pregnant ewes

Seventeen cases of spontaneous, partial or total vaginal rupture, in pregnant ewes, involving the dislocation and herniation of the intestines and uterus, were studied. Four of them also had a uterine torsion, and three of these recovered after treatment. In the remaining 13 cases the condition of the uterus was unknown. The lesion always consisted of a dorsolateral tear in the vagina with a partial or total perforation of the wall close to the uterine cervix. The affected animals were all in normal body condition. Their average age was just under five years, and most were carrying twins. Most cases occurred approximately one week before expected lambing. None of the cases was observed to have a vaginal prolapse before the vaginal rupture. Histological examination of one case revealed scar formation in the vaginal wall close to the rupture, which appeared to be due to an earlier inflammatory process or injury. The circulatory disturbance in the reproductive organs caused by the uterine torsion potentially weakens the vaginal wall. This weakness, in combination with excessive tenesmus resulting from increased tension in the uterine ligaments, and in some cases possibly with a lower vaginal resistance due to previous scarring, may be of aetiological significance in spontaneous vaginal rupture.     

A pharmacokinetic study including some relevant clinical effect of medetomidine and atipamezole in lactating dairy cows

Medetomidine is the most potent and selective alpha2-agonist used in veterinary medicine and its effects can be antagonized by the alpha2-antagonist atipamezole. The pharmacokinetics of medetomidine and atipamezole were studied in a cross-over trial in eight lactating dairy cows. The animals were injected intravenously (i.v.) with medetomidine (40 microg/kg) followed by atipamezole i.v. (200 microg/kg) or saline i.v. after 60 min. Drug concentrations in plasma were measured by HPLC. After the injection of atipamezole, the concentration of medetomidine in plasma increased slightly, the mean increment being 2.7 ng/mL and the mean duration 12.1 min. However, atipamezole did not alter the pharmacokinetics of medetomidine. It is likely that the increase in medetomidine concentration is caused by displacement of medetomidine by atipamezole in highly perfused tissues. The volume of distribution at steady state (Vss) for medetomidine followed by saline and medetomidine followed by atipamezole was 1.21 and 1.32 L/kg, respectively, whereas the total clearance (Cl) values were 24.2 and 25.8 mL/min x kg. Vss and Cl values for atipamezole were 1.77 mL/kg and 48.1 mL/min x kg, respectively. Clinically, medetomidine significantly reduced heart rate and increased rectal temperature for 45 min. Atipamezole reversed the sedative effects of medetomidine. However, all the animals, except one, relapsed into sedation at an average of 80 min after injection of the antagonist.     

A retrospective evaluation of a Bovine Herpesvirus-1 (BHV-1) antibody ELISA on bulk-tank milk samples for classification of the BHV-1 status of Danish dairy herds

Bulk-tank milk samples analysed in a Bovine Herpesvirus-1 (BHV-1) blocking ELISA are still in use in the Danish BHV-1 programme as a tool to classify dairy herds as BHV-1 infected or BHV-1 free herds. In this retrospective study, we used data from the Danish BHV-1 eradication campaign to evaluate performance characteristics of the BHV-1 blocking ELISA in 1039 BHV-1-seropositive and 502 repeatedly BHV-1-negative dairy herds using the results of blood testing of the individual animals as the true infection status. At a cut-off value of 30% blocking reaction, the herd-level relative sensitivity and relative specificity were 82 and 100%, respectively. The herd-level relative sensitivity depended on the within-herd prevalence of seropositive cows and the cut-off value in the assay, but not on the time interval (up to 90 days) between the collection of the bulk-tank milk sample and the individual serum samples. The BHV-1 blocking ELISA on bulk-tank milk could detect seropositive herds (few), with prevalence proportions as low as one seropositive cow out of 70 cows.     

Studies on the effects of intravenous administration of glucose, fructose, invertose and sorbitol on various blood constituents of blood plasma (monosaccharides, insulin, lactate, pyruvate and free fatty acids as well as glutamate-oxaloacetate transaminase) in the horse

Horses were examined for the behaviour of various blood constituents prior to and following infusions of solutions of glucose, fructose, invertose, and sorbitol. Infusion of 0.5 g/kg live weight glucose to six horses was followed by half-life variation between eleven and 23 minutes. Subsequent infusion of invertose to the same animals usually caused prolongation of glucose half-life. Half-life values were between 17 and 33 minutes for fructose and between 21 and 80 minutes for glucose. Infusion of 0.5 g/kg live weight fructose to two horses was followed by half-life values between 17 and 18 minutes, while the half-life values of sugar alcohol were 16, 16, 27, and 29 minutes in four horses who had received sorbitol. Sugar or sorbitol infusion was not followed by substantive change of lactate and pyruvate concentrations in the blood or free fatty acids in the blood plasma or GOT activity. The rise of insulin in the blood plasma was differentiated. Invertose and sorbitol solutions, consequently, can be recommended for application to horses.