Use of carbendazim and carbendazim-resistant yeasts to create different yeast densities on wheat leaves for field studies on biological control

Phyllosphere yeasts may play an important role in suppressing necrotrophic pathogens. In a randomized block design, yeast densities on flag leaves and second leaves of field-grown wheat were left unchanged (D), or were lowered by sprayings with carbendazim (A), or were raised by sprayings with a mixture ofSporobolomyces roseus, Cryptococcus laurentii var.flavescens and nutrients (B), or with carbendazim, carbendazim-resistantS. roseus andC. laurentii var.flavescens and nutrients (C). The application of carbendazim to plots A and C would avoid a biased interference of carbendazim due to other effects than reducing the natural yeast population. Prolonged differences in yeast density would be reflected in differences in severity of diseases caused by carbendazim-insensitive necrotrophic pathogens.Both treatments B and C enhanced the yeast density about 10-fold at the beginning of leaf colonization, 2 to 3 weeks after leaf emergence. Untreated leaves (D), however, were rapidly leaves. Initially, the yeasts were suppressed by carbendazim (A). However, progressively the yeast population consisted of carbendazim-resistant strains which made the carbendazim sprayings ineffective. Therefore, the substantial differences in yeast density were of limited duration. The effect of the treatments on naturally occurringMycosphaerella graminicola (anam.Septoria tritici), Puccinia recondita f. sp.tritici and the total necrotic leaf area (D-value) is discussed.Samenvatting Fyllosfeergisten kunnen misschien een belangrijke rol spelen in de onderdrukking van necrotrofe pathogenen. In een blokkenproef werd de dichtheid van gisten op vlagbladeren en tweede bladeren niet beïvvloed (D) of verlaagd door bespuitingen met carbendazim (A), of verhoogd door bespuitingen met een mengsel van gisten (Sporobolomyces roseus enCryptococcus laurentii var.flavescens en voedingsstoffen (B), of met een mengsel van carbendazim, carbendazim-resistenteS. roseus enC. laurentii var.flavescens en voedingsstoffen (C). De grootste verschillen in dichtheden van de gistpopulaties werden verwacht tussen de behandelingen A en B of C. Door alle veldjes A en C met carbendazim te behandelen wordt een eenzijdig effect van carbendazim, berustend op andere eigenschappen dan de onderdrukking van de natuurlijke gistpopulatie, voorkomen. Langdurige verschillen in de populatiedichtheden van de gisten zouden weerspiegeld kunnen worden in verschillen in aantasting door carbendazimongevoelige necrotrofe pathogenen.Aan het begin van de bladkolonisatie door gisten, 2 tot 3 weken na het verschijnen van het blad, leidde zowel behandeling B als C tot, een 10-voudige verhoging van de gistdichtheid. De onbehandelde bladeren (D) werden echter spoedig door de van nature voorkomende gisten gekoloniseerd en het verschil tussen de onbehandelde en met gist bespoten bladeren werd snel kleiner. Aanvankelijk werden de gisten door carbendazim (A) onderdrukt. Geleidelijk bestond de gistpopulatie echter uit carbendazim-resistente stammen, zodat de carbendazim bespuitingen nauwelijks meer effect hadden. Hierdoor werden grote verschillen in gistdichtheden niet langdurig gerealiseerd. Het effect van de behandelingen op de natuurlijke infectie doorMycosphaerella graminicola (anam.Septoria tritici), Puccina recondita f. sp.tritici en op het totale dode bladoppervlak (D-waarde) wordt besproken.     

Evaluation of Microbial Antagonists for Biological Control of Botrytis cinerea Stem Infection in Cucumber and Tomato

Of fifteen isolates of yeasts, filamentous fungi and bacteria and a commercial product, tested in a bioassay with stem segments, eleven isolates consistently reduced incidence of disease and sporulation of Botrytis cinerea Pers; Fr in tomato and seven isolates in cucumber. Several isolates reduced disease by more than 75% in all experiments. Six antagonists that performed well in the bioassays and that were fairly easy to produce in vitro, were selected for further testing in two glasshouse experiments with cucumbers. After application of spores of B. cinerea and the antagonists or the fungicide tolylfluanid to pruning wounds, disease incidence was reduced by 50–100% by all antagonists in both experiments and only in one experiment by tolylfluanid.For Trichoderma harzianum T39, Aureobasidium pullulans and Cryptococcus albidus, biological control efficacy in bioassays with cucumber stem segments was not strongly influenced by temperatures in the range between 18 and 30°C, but at 24°C the efficacy of the three antagonists strongly decreased at relative humidities of 90% and 80% (vapour pressure deficits 0.299 and 0.598kPa, respectively) compared to 100.     

RADIOGRAPHY OF THE EQUINE STOMACH

To obtain radiographic information concerning the equine stomach, a gastrographic contrast examination is required. This study describes this procedure in detail. A powerful radiographic unit, the tubehead linked to an image intensifier and suspended by an electromechanical overhead gantry system, is required. To obtain accurately positioned radiographs during the fluoroscopic examination, a cassette holder with a stationary grid is mounted at the entrance window of the image intensifier. The examination is performed in the unsedated standing horse after 24 hours of starvation, using a combination of survey radiography and fluoroscopic viewing after the inflation of air, followed by the administration of barium sulphate suspension by stomach tube. The gastrographic contrast examination is performed in three experimental animals and 23 abnormal horses. Pneumogastrophy appeared to be valuable to diagnose gastric tumors, to differentiate between gastric tumors and other masses in the cranial abdomen, and to visualize gastric parasites, even in large horses. The use of barium sulphate suspension does not result in an adequate double contrast of the stomach, but it may aid to diagnose esophagogastric or pyloric stenosis and gastric or duodenal ulcers.     

Lag screw fixation of sagittal navicular bone fractures in five horses

A sagittal fracture of the navicular bone in five horses was treated by means of lag screw. Intra-operative radiographic monitoring and a specially developed guide apparatus was necessary to implant the screw precisely along the transverse axis of the navicular bone. Internal fixation resulted in progressive radiographic narrowing of the fracture zone. In all five patients the fracture healed (bony union) without superfluous callus formation.     

Heredity of navicular disease

Bone structure, nutrient canals, form and definition, and a total evaluation of both fore navicular bones were radiologically evaluated in 169 three- and four-year-old Dutch warmblood mares divided into eleven daughter (half sister) groups. On the basis of the 'total evaluation', differences between the three- and four-year-old horses were detectable. Using the 'total evaluation', significant differences were found between daughter groups consisting of three-year-old mares. These differences support the theory that podotrochleosis may be (partly) due to the presence of genetic factors. The possibilities concerning selection are presented. The results of the radiological evaluation of the stallion do not have predictive value as far as his progeny is concerned.     

The arterial supply of the navicular bone in adult horses with navicular disease

A macroscopical, arteriographical and histological study was made of the arterial supply of the navicular bones of horses with clinical and/or radiographic signs of navicular disease. Based on the clinical and radiological findings the navicular bones of 40 Warmblood horses are divided into four different groups. In the pattern of the arteriogram obvious changes are noticed: an imbalance and a distal to proximal shift between the distal and proximal arterial supply occur, indicating a reduction of the distal blood supply with a compensatory reaction of the proximal, medial and lateral supply. The changes in the pattern of the arteriogram are histologically defined by arterio(lo) sclerosis and newly formed arteries. The presence of radiologically visible nutrient foramina is associated with a changed pattern in the arteriogram, increased bone remodelling and fibrosis. Ischaemia and increased pressure (hypertension and/or increased intra-articular pressure) are considered to be responsible for changes in form and number of radiologically visible nutrient foramina.     

The arterial supply of the navicular bone in the normal horse

An arteriographic and histological study of the arterial blood supply of 35 navicular bones, from 19 adult horses with no clinical or radiographical signs of navicular disease, is described. The arteries enter the navicular bone from four directions: distal, proximal, medial, and lateral. The navicular bone can be divided into areas which receive arteries from one, two, or three directions, whereas the distal supply covers the largest part of the navicular bone. The distal arteries enter the navicular bone through the nutrient foramen which contains loose connective tissue, nutrient vessels and myelinated nerves. At least 77 per cent of the nutrient foramina contains synovial membrane. Twelve per cent of the arteries in the navicular bone show histological changes. Thickening of the intima with or without splitting of the internal elastic membrane and hyalinization, fibrosis and hypertrophy of the media were observed.     

Haematogenous osteomyelitis in cattle

The examination of 70 cattle with haematogenous osteomyelitis resulted in the classification of the bone lesions into two main groups: the physeal type, in which an infection, usually of metaphyseal bone, originated at or near the growth plate, usually in the distal metacarpus, metatarsus, radius or tibia, and the epiphyseal type, in which an infection originated near the junction of the subchondral bone and the immature epiphyseal joint cartilage, most often in the distal femoral condyle epiphysis, the patella and the distal radius. Combinations of physeal and epiphyseal defects and even diaphyseal involvement were occasionally seen. Epiphyseal osteomyelitis was mostly caused by salmonella infection, physeal by Corynebacterium pyogenes, salmonella and other bacteria. The salmonella affected animals were with one exception less than 12 weeks old and the majority had had some previous illness or came from a problem herd. The C pyogenes affected calves were in almost all cases more than six months old. The prognosis of the metaphyseal infection was in general satisfactory, and surgical intervention (osteotomy or sequestrectomy) was often required. The prognosis of the epiphyseal type was grave but two of the three animals in which physeal and epiphyseal defects were accompanied by diaphyseal lesions recovered.     

Tarsal osteomyelitis in foals

Nine foals with haematogenous osteomyelitis of the tarsal bones were studied clinically, radiologically and pathologically. Clinical examination, including aspiration of articular or periarticular effusions in the tarsal area, and radiological study provide definitive diagnosis in most cases.     

External quality assurance system for antibiotic resistance in bacteria of animal origin in Europe (ARBAO-II), 2003

Initiated in 2003 by the European Union, ARBAO-II aims to establish a monitoring of antimicrobial susceptibility among the veterinary laboratories in all European countries based on validated methodologies. This includes an external quality control system for the most important bacterial pathogens. In 2003 two trials were performed. The first on Salmonella and Escherichia coli involved 31 laboratories in 18 countries and the second on Staphylococcus and Streptococcus included 22 laboratories in 15 countries. For the E. coli strains, 92.8% of the results complied with the reference MICs, for Salmonella 93.7%, for Streptococcus 80.4% and for Staphylococcus 93.1%. Most problems were observed when testing florfenicol (79.2% correct), gentamicin (84.2%) and amoxicillin + cl (84.9%) in E. coli; streptomycin in Salmonella (62.5%); gentamicin (56.7%), lincomycin (71.4%), clindamycin (75.4%), TMP + sulfonamides (75.7%) and chloramphenicol (78.5%) in Streptococcus; erythromycin (81.5%) and oxacillin (78.2.5%) in Staphylococcus. A few laboratories caused most deviations. However, there was no correlation between good performance for one bacterial group and good performance for other groups. This study showed that most laboratories are capable of performing correct susceptibility testing for E. coli and Salmonella, even though performance of some laboratories can be improved, and that some problems exist for Staphylococcus and Streptococcus. This clearly shows the need for continuous harmonisation of methodologies within the EU.