Evaluation of benomyl and thiophanate-methyl for the control of Verticillium wilt of strawberry in the Netherlands

In pot experiments the fungicides benomyl and thiophanate-methyl controlledVerticillium wilt of strawberry when applied as a soil drench after planting. Both compounds were ineffective as foliar sprays and as root dips prior to planting. Soil drenches applied to commercially grown runner plants in the waiting field (August) and to the same plants in the greenhouse (December or January) increased the yield. On infested ground, a soil drench with thiophanate-methyl promoted the occurrence of crown rot caused byPhytophthora cactorum.     

Detection of viral antigen in semi-thin sections of plant tissue by immunogold-silver staining and light microscopy

The immunogold-silver staining technique was developed for the light microscopical localization of viral antigen in plant tissue. Semi-thin sections of LR White-embedded plant tissue were immunologically labelled with primary antiserum and protein A-gold. Individual gold particles were covered with a silver precipitate using a physical developer. This precipitate could be seen as black spots in a conventional light microscope with brightfield and as brilliant white spots with darkfield illumination. Maximal sensitivity and low background was obtained when immunogold-labelled sections were fixed in glutaraldehyde prior to silver enhancement. Simultaneous observation of the silver coated gold label and cell morphology was achieved by epipolarization microscopy. Using this technique cowpea chlorotic mottle virus coat protein was detected in cowpea plants as function of the infection period. Virus translocation and multiplication was monitored in systemically inoculated tissue, showing viral antigen in phloem parenchyma of petiolules 6 h after systemic inoculation and subsequent spreading from the phloem to the neighbouring bundle sheath and cortex cells.Samenvatting De immunologische techniek (IGSS), waarbij complexen van antilichamen met proteïne A geadsorbeerd aan kolloïdaal goud (pAg) worden bedekt met zilver, werd met succes toegepast voor het aantonen van viraal antigeen in geïnfecteerd planteweefsel met behulp van de lichtmicroscoop. Semi-dunne plakjes weefsel werden ingebed in LR White en behandeld met antiserum tegen het cowpea chlorotic mottle virus (CCMV). Aan dit antigeen-antilichaam complex werd pAg gehecht. Vervolgens werd op de individuele gouddeeltjes zilver neersgeslagen met een ontwikkelaar bestaande uit een mensel van zilverlactaat en hydroquinone. De gouddeeltjes katalyseren de reductie van de zilverionen in oplossing tot metallisch zilver, dat neerslaat op de gouddeeltjes. Het zilverprecipitaat is waarneembaar als zwarting in een lichtmicroscoop met doorvallend licht en licht wit op bij donkerveld belichting. Maximale gevoeligheid van detectie en lage achtergrondkleuring werden bereikt door fixatie van het antigeen-antilichaam-pAg complex met glutaaraldehyde vóór de zilverkleuring.Gelijktijdige waarneming van het zilver label en de morfologie van de cellen was mogelijk door toepassing van gepolarisserd licht in een microscoop met opvallende belichting (epipolarisatiemicroscopie) in combinatie met doorvallend licht. Het zilverprecipitaat is hierbij waarneembaar als een helder blauwe kleur door de weerkaatsing en verstrooiing van het gefiltreerde gepolariseerde licht, terwijl de morfologie van de cytochemisch gekleurde cellen zichtbaar is met doorvallende belichting. Met IGSS in combinatie met epipolarisatiemicroscopie werd het CCMV gelokaliseerd in cowpea planten als functie van de infectieduur. De translocatie en vermenigvuldiging van het virus werden gevolgd in planteweefsel dat systemisch was geïnoculeerd volgens de differentiële-temperatuur-inoculatietechniek. Zes uur na systemische inoculatie werd het virus voor het eerst waargenomen in enkele floeemparenchymcellen en de infectie breidde zich daarna snel uit. Vierentwintig uur na inoculatie kon virus worden aangetoond in grote delen van het floeem, in de bundelschede en in de aangrenzende delen van de schors. Concluderend kan worden gesteld dat het virus vanuit de primaire bladeren naar de secundaire bladeren werd getransporteerd via het floeem, analoog aan het transport van assimilaten.Tot slot werden dunne plakjes geïnfecteerd weefsel, geïncubeerd met antiserum en proteïne A-goud, na zilverbehandeling vergeleken in licht-en elektronenmicroscoop.     

Removing faecal contamination of broilers by spray‐cleaning during evisceration

1. The faecal contamination of broiler carcasses during evisceration results in an increase in contamination with Enterobacteriaceae, including any salmonellas present.

2. This increase can be prevented completely by spray‐cleaning carcasses during the various stages of evisceration.

3. If the carcasses are cleaned only at the end of the evisceration process, the numbers of Enterobacteriaceae are not reduced to initial levels and Salmonella contamination is less efficiently removed.     

Characterization of a carlavirus in elderberry (Sambucus spp.)

A carlavirus was isolated fromSambucus racemosa andS. nigra in the Netherlands. The virus was sap-transmissible and capable of infecting 14 out of 58 plant species and cultivars tested, causing symptoms in five of them. It was also transmitted byMyzus persicae at a low rate. Dilution end-point was 10^–3–10^–4, thermal inactivation at 70–75°C and ageing in vitro 2–4 days. The virus had a sedimentation coefficient of 155 S and molecular weight of capsid protein subunits of 31 000 dalton. The average buoyant density of the four isolates used was 1.315 g/cm³. The virus particles had an average normal length of 678 nm and a width of approximately 12 nm. In ultrathin sections of leaf tissue ofS. racemosa Plumosa Aurea bundles of virus particles were observed in the cytoplasm. Close serological relationship was found to a virus isolated from elderberry in Britain and a distant relationship to carnation latent virus. In its reaction on host plants and its persistence in crude sap it also resembled the former virus, originally code-named elderberry virus A. We propose the name elderberry carlavirus for it.Samenvatting Een carlavirus werd geïsoleerd uitSambucus racemosa enS. nigra in Nederland. Het virus kon met sap worden overgebracht en was in staat 14 van de 58 getoetste plantesoorten en-cultivars te infecteren waarbij op vijf van deze symptomen verschenen. Ook metMyzus persicae vond overdracht plaats, zij het in beperkte mate. De verdunningsgrens was 10^–3–10^–4, de inactiveringstemperatuur 70–75°C en de houdbaarheid in vitro 2–4 dagen. Het virus had een sedimentatiecoëfficiënt van 155 S en het molecuulgewicht van de structuurelementen van het capside-eiwit bedroeg 31 000 dalton. De deeltjes van de vier gebruikte isolaten hadden een gemiddelde zweefdichtheid van 1,315 g/cm³. De gemiddelde normale lengte van de virusdeeltjes bedroeg 678 nm bij een breedte van ongeveer 12 nm. In ultradunne coupes van bladweefsel vanS. racemosa Plumosa Aurea werden bundels draadvormige virusdeeltjes waargenomen in het cytoplasma. Het virus vertoonde een zeer sterke serologische verwantschap met een virus uit vlier geïsoleerd in Groot-Brittannië en een geringe verwantschap met het anjer-latenvirus. In zijn reactie op waardplanten en zijn eigenschappen in ruw sap vertoonde het ook veel gelijkenis met eerstgenoemd virus, in de literatuur vermeld onder de code-naam elderberry virus A. We stellen voor de naam carlavirus van vlier aan dit virus te geven.     

Molecular breeding for virus resistant potato plants

To engineer resistance against potato virus X (PVX), the viral coat protein (CP) gene has been introduced into two potato cultivars. Stable expression of the gene in transgenic clones throughout the growing season has been obtained and resulted in considerably increased virus resistance. With varying frequencies depending on the original cultivar used, true-to-type PVX resistant transgenic clones have been obtained. Since deviant light sprout characteristics were invariably associated with aberrations in plant phenotype, they can be used in procedures to early screen for deviations. Furthermore, it has been possible to unequivocally discriminate between the original untransformed and independent transgenic cultivars. Although no relation has been found between the presence, if any, of the CP of potato virus Y (PVY) or potato leafroll virus (PLRV) in CP gene transgenic potato, appreciable levels of resistance to these viruses has been obtained. This suggests that the mechanism by which a viral CP gene in the potato genome evokes resistance, differs amongst various viruses.     

Feed intake and rumen motility in dwarf goats. Effects of some α-adrenergic agonists,prostaglandins and posterior pituitary hormones

The purpose of the present investigation was to test the hypothesis that drug-induced changes in rumen contractions influence feed intake in dwarf goats. Intravenous (i.v.) administration of clonidine (1 g kg^-1 min^-1 for 10 min), xylazine (1 g kg^-1 min^-1 for 10 min), and PGF-2 (10 g kg^-1 min^-1 for 15 min) caused bradycardia and inhibition of rumen contractions. However, no appetite-stimulating effect of these drugs was observed. Other clinical changes induced by the ₂-adrenergic agonists included slight sedation and a decrease in body temperature; all clinical effects of clonidine and xylazine were partly antagonized by tolazoline pretreatment (10 g kg^-1 min^-1 for 30 min). These results suggest that the CNS control of feeding differs in ruminants and monogastric species.In dwarf goats fasted for 2 h, i.v. administration of oxytocin (0.01 IU kg^-1 min^-1 for 15 min), vasopressin (0.01 IU kg^-1 min^-1 for 15 min), octapressin (0.003 IU kg^-1 min^-1 for 15 min) or PGE (0.8 g kg^-1 min^-1 for 15 min) did not change feeding behaviour during the two observation periods (0–30 min and 180–210 min after drug infusion, respectively). In previous studies, similar doses of these drugs induced changes in heart rate and inhibition of rumen contraction in goats. These findings demonstrate that drug-induced changes in forestomach contractions do not simply cause changes in feeding behaviour. The i.v. infusion of the PGF₂ analogues etiproston (10 g kg^-1 min^-1 for 15 min), luprostiol (30 g kg^-1 min^-1 for 15 min), cloprostenol (1 g kg^-1 min^-1 for 15 min) and tiaprost (1 g kg^-1 min^-1 for 15 min) induced hypophagic effects and stimulated intestinal propulsion.     

Comparative serological response in calves to eight commercial vaccines against infectious bovine rhinotracheitis, parainfluenza-3, bovine respiratory syncytial, and bovine viral diarrhea viruses

A field trial was conducted to compare the serological responses in calves to eight commercial vaccines against infectious bovine rhinotracheitis virus (IBRV), parainfluenza-3 virus (PI3V), bovine respiratory syncytial virus (BRSV), and/or bovine viral diarrhea virus (BVDV). Calves given IBRV, P13V, BRSV, and BVDV vaccines had significantly higher antibodies to these viruses than unvaccinated controls; however, serological responses to killed BVDV vaccines were low. Calves with preexisting antibodies to IBRV, PI3V, BRSV, and the Singer strain of BVDV had lower seroconversion rates following vaccination than calves that were seronegative initially.

Serological responses in calves to IBRV, PI3V, BRSV, and BVDV differed among various commercial vaccines. Antibody titers to IBRV were higher in calves vaccinated with modified-live IBRV vaccines than in those vaccinated with killed IBRV vaccines. Following double vaccination with modified-live IBRV and PI3V vaccines, seroconversion rates and antibody titers to IBRV and PI3V were higher in calves vaccinated intramuscularly than in those vaccinated intranasally. Calves given Cattlemaster 4 had significantly higher titers to BRSV and PI3V, and lower titers to BVDV, than calves given Cattlemaster 3, suggesting that the addition of BRSV to Cattlemaster 4 caused some interaction among antigens.