Specific detection of Histomonas meleagridis in turkeys by a PCR assay with an internal amplification control

Histomoniasis or blackhead is a disease of gallinaceous birds, caused by the protozoon Histomonas meleagridis. Since traditional diagnostics for the detection of this disease are complex and far less sensitive than molecular tools, a PCR would provide a more rapid and sensitive alternative. However, intestinal material and droppings, which are preferably used in epidemiological studies of histomoniasis, often contain PCR inhibitory substances. To detect these false negative results, the use of an internal amplification control is essential. Nevertheless, the recently developed PCR tests lack this internal control. Therefore, a new PCR assay with H. meleagridis specific primers was developed which does include an internal amplification control. The diagnostic value of the PCR assay was evaluated in comparison to three other conventional H. meleagridis specific PCR tests (HIS5, HM1 and HM2). None of the organ samples originating from uninfected turkeys, showed positive PCR results in any of the tests. Among the lesion-positive, inhibition-free samples, 95.4% were positive by our PCR assay, while only 50, 66.7 and 83.3% of the lesion-positive organs tested positive by the HM1, the HIS5 and the HM2 PCR respectively. In conclusion, our PCR offers the use of the internal control to detect false negative results and an increased sensitivity, and thus should be useful for routine diagnosis of H. meleagridis in poultry.     

Antiapoptotic activity of bovine herpesvirus type-1 (BHV-1) UL14 protein

Viruses have evolved different strategies to interfere with apoptotic pathways in order to halt cellular responses to infection. One previous study showed that transient transfection of bovine herpesvirus type-1 (BHV-1) UL14 protein is efficient in protecting Madin Darby kidney (MDBK) and human chronic myelogenous leukemia (K562) cells from sorbitol-induced apoptosis. This protein corresponds to a putative protein of BHV-1, which shares aminoacid sequence with a part of the peptide-binding domain conserved in human heat shock protein (HSP70) family. The pBK-CMV-UL14 plasmid transfected MDBK cells treated with sorbitol did not show caspase-3 and caspase-9 activation with respect to non-transfected MDBK cells (UL14 negative). Furthermore, we report that the expression of the full length sequence of BHV-1 UL14 is evident after 7 h of infection of BHV-1 on MDBK cells which were then treated with sorbitol. These results indicate that UL14 gene product has important implications to enhance cell survival in response to apoptotic stimuli.     

Incidence of Salmonellae in Captive and Wild Free‐Living Raptorial Birds in Central Spain

A total of 595 faecal samples from raptorial birds, either captive or free‐living, residing in GREFA Wildlife Hospital were bacteriologically examined using various selective media and an Automated Diagnostic Assay System for Salmonella detection. Serotype and phage type of the strains identified as Salmonella was determined. In the captive group, of the 285 samples examined, 21 (7.36%) were positive for Salmonella. Serotyping revealed that most of the individuals were infected by Salmonella serotype Havana. This result suggested that there could be a source of contamination in the Hospital although it could not be established. In the wild free‐living group, over 310 samples examined (4.19%) were positive for Salmonella. The Salmonella isolates showed a major variety of serotypes: Enteritidis, Adelaide, Brandenburg, Newport, Typhimurium, Hadar, Saintpaul and Virchow. Most of them are similar to those commonly described in isolates from human and domestic animals. These results indicate that wild birds could be involved in the dissemination of Salmonella in humans or domestic animals or vice versa.     

Detection of early embryonic development in chicken eggs using visible light transmission

1. In two separate experiments, the possibility of detecting embryonic development in chicken eggs was assessed using the same spectrophotometric method used to detect blood in Table eggs, using a combination of two wavelengths (577 and 610 nm) of the transmission spectrum. 2. In the first experiment, during the first 10 d of incubation, transmission spectra of 30 Hisex White eggs and 30 Hybro eggs were measured daily. 3. In the second experiment, 292 Hisex White eggs were incubated. Seven groups were randomly assigned. Six received an injection of sodium azide (NaN3) at different times during incubation in order to stop embryonic development, and during the first 12 d of incubation the transmission spectrum was measured daily. The acoustic resonance analysis method was also used on a group of uninjected eggs. 4. In the first experiment, it was possible to detect embryonic development from 120 h of incubation onwards in fertile eggs. In the second experiment changes in light transmission due to embryonic development were detected from 108 h of incubation. Detection of embryonic development using the acoustic resonance analysis method in the second experiment was possible only from 120 h of incubation. 5. It was concluded that the detection of embryonic development using visible light transmission is not directly linked with the formation of blood, but with the formation of sub-embryonic fluid, which takes place from 72 h of incubation onwards. This fluid makes the yolk sac translucent so that absorption of light at 577 nm can be detected.     

The gamma-interferon test: its usefulness in a bovine tuberculosis survey in African buffaloes (Syncerus caffer) in the Kruger National Park

A survey to determine the bovine tuberculosis status of buffalo herds north of the Olifants River in the Kruger National Park was conducted, using a new diagnostic approach. Diagnosis of Mycobacterium bovis infection was accomplished using the gamma-interferon assay technique in 608 adult buffaloes out of a total of 29 discreet herds. The animals were immobilized in groups of 10-15, bled, individually marked and then revived and released on site. As soon as test results were available (after 26-36 h), the same buffalo herd was relocated by tracking the frequency of a radio-collar previously fitted to one adult cow per group during the initial operation. Bovine reactors were identified, darted and euthanased from the helicopter. Necropsy and culture findings of all culled buffaloes showed excellent correlation with the results of the ante-mortem gamma-interferon test. The survey revealed that over and above the two positive herds that had been identified during a previous survey carried out in 1996, there were three additional, but previously unidentified, infected herds in the region north of the Olifants River.     

Comparison of nuclear morphometric parameters in cytologic smears and histologic sections of spontaneous canine tumors

Background — Nuclear morphometry may provide useful diagnostic and prognostic information for neoplasms in animals. Most available data have been obtained from histologic sections. Nuclear morphometry of cytologic smears may provide important pre-operative information.
Objectives — The goal of this study was to compare nuclear morphometric parameters in cytologic smears and histologic sections from spontaneous canine tumors.
Methods — Mean nuclear area (MNA), mean nuclear perimeter (MNP), mean nuclear form factor (FF; nuclear perimeter²/4π nuclear area) and their respective SDs were assessed by image analysis of both hematoxylin and eosin-stained histologic sections and May-Grünwald-Giemsa-stained cytologic smears from the same case in 20 spontaneous canine tumors of different histogenesis. The above parameters were selected as being the best morphometric tools for measuring variation in shape and size in cells after neoplastic transformation. Data were compared by ANOVA with P<.01 considered significant.
Results — There was a significant difference between histologic and cytologic specimens for MNA, MNP, and their SDs. Only the differences between FF and the SD of FF were not statistically significant.
Conclusions — Only nuclear morphometric data related to nuclear shape and nuclear shape variability are comparable between histologic and cytologic specimens. Nuclear area and perimeter may be affected by the different fixation and smear preparation techniques used in histology and cytology.     

Tumours of Begonia and some other ornamentals,induced by Corynebacterium fascians

In 1975 many tumours were observed in plants ofBegonia Schwabenland grown in Aalsmeer. Submersion of the roots ofNicotiana megalosiphon seedlings in a homogenate of tumorous tissue, induced tumours after two weeks. Short periods of submergence yielded results similar to those obtained after longer periods. Tumour homogenates lost their infectivity after ten min at 50°C. Aphids transmitted the infectious agent.Treatment with propylene oxide did not inhibit infectivity completely. Filtration through a 450 nm filter removed the infectious agent.Tobacco tumor virus or a viroid could not be isolated. Cultures ofCorynebacterium fascians, isolated from tumours ofN. megalosiphon were highly infectious and induced tumours in healthyN. megalosiphon andBegonia. Tumorous tissue homogenates ofPelargonium zonale, Dahlia sp.,Gladiolus sp., andLilium sp. also caused tumours inN. megalosiphon, from whichC. fascians was isolated. It was not possible to produce tumours inN. megalosiphon with homogenates from roses with symptoms of bud proliferation.Samenvatting In 1975 werden vele tumoren waargenomen inBegonia Schwabenland op Aalsmeerse bedrijven (Fig. 1). De infectiositeit van tumorweefsel kon goed en snel worden vastgesteld door de wortels van zaailingen vanNicotiana megalosiphon in een homogenaat van tumorweefsel te dompelen. Tumoren ontstonden na twee weken, de eindbeoordeling geschiedde na een maand (Fig. 2). Ook verschillende andereNicotiana spp.,Melilotus officinalis (Fig. 3) enPisum odoratum (Fig. 4) werden aangetast.Bij de infectiositeitstoets gaven zeer korte dompeltijden even goede resultaten als langere (Tabel 1). Infectieus sap verloor zijn infectievermogen na 10 min verhitting bij 50°C. Bladluizen brachten de smetstof over. Propyleenoxide verminderde de infectiositeit wel, doch onderdrukte deze niet totaal. Bij filtratie door een 450 nm filter bleef het infectieuse agens op het filter achter. Het tumor-inducerende agens was ook aanwezig in die delen van planten met tumoren welke gezond leken en het ging voor een gering deel over met zaad (Tabel 2).Uit tumoren konden wij geen tabakstumorvirus of een viroïde isoleren. Culturen vanCorynebacterium fascians, geïsoleerd uit tumoren vanN. megalosiphon bleken zeer infectieus en veroorzaakten tumoren inN. megalosiphon enBegonia. Homogenaten van tumorweefsel vanPelargonium zonale, dahlia (Fig. 5), gladiool (Fig. 6) enLilium Mid Century Hybrid Enchantment (Fig. 7) veroorzaakten ook tumoren opN. megalosiphon, waaruitC. fascians werd geïsoleerd. Met sap van kroeskopzieke rozen konden wijN. megalosiphon niet besmetten.     

Determination and persistence of imazalil in post-harvest-treated apples during cold storage

A method to determine imazalil (allyl 1 -(2, 4-dichlorophenyl)-2-imidazol- 1-ylethyl ether) on apples using high performance liquid chromatography is described. After harvest, fruits were immersed in an aqueous suspension (1 g litre-1) and cold stored (0-2°C) at 85-90% r.h. Samples were taken monthly and imazalil was determined in the peel, outer pulp, inner pulp and on whole fruit. Residues were 3.8-4.9 mg imazalil kg-1 and decreased during storage. Most imazalil was found on the peel and amounts in the pulp decreased toward the core.     

Perceptions of risk, risk aversion, and barriers to adoption of decision support systems and integrated pest management: an introduction

Rational management of plant diseases, both economically and environmentally, involves assessing risks and the costs associated with both correct and incorrect tactical management decisions to determine when control measures are warranted. Decision support systems can help to inform users of plant disease risk and thus assist in accurately targeting events critical for management. However, in many instances adoption of these systems for use in routine disease management has been perceived as slow. The under-utilization of some decision support systems is likely due to both technical and perception constraints that have not been addressed adequately during development and implementation phases. Growers' perceptions of risk and their aversion to these perceived risks can be reasons for the "slow" uptake of decision support systems and, more broadly, integrated pest management (IPM). Decision theory provides some tools that may assist in quantifying and incorporating subjective and/or measured probabilities of disease occurrence or crop loss into decision support systems. Incorporation of subjective probabilities into IPM recommendations may be one means to reduce grower uncertainty and improve trust of these systems because management recommendations could be explicitly informed by growers' perceptions of risk and economic utility. Ultimately though, we suggest that an appropriate measure of the value and impact of decision support systems is grower education that enables more skillful and informed management decisions independent of consultation of the support tool outputs.