The use of monoclonal antibodies in an enzyme immunospot assay to detect isotype-specific antibody-secreting cells in pigs and chickens

Monoclonal antibodies directed against porcine immunoglobulin isotypes G, G1, G2, M, and A and against chicken immunoglobulin isotopes G, M, and A were tested in an antigen-specific spot-forming cell (SFC) assay based on the principle of the enzyme immunoassay. The SFC assay was used to quantitate ovalbumin (OA)-specific antibody-secreting cells (ASC) in pigs that had been primed and boosted with OA. The SFC assay was also used to quantitate trinitrophenyl (TNP)-specific ASC in chickens that had been primed with TNP-conjugated keyhole lympet haemocyanin (TNP-KLH). Although, the classical plaque-forming cell (PFC) assay cannot reliably detect isotope-specific ASC in pigs and chickens, it can detect these cells in mice. Therefore, we compared the OA- and TNP-specific SFC assays with PFC assays that were specific for these antigens in mice. The study demonstrated that the SFC assay is superior to the PFC assay in detecting both OA-specific ASC and TNP-specific ASC. The frequencies of OA-specific and TNP-specific SFC detected in mice were of the same order of magnitude as those detected in pigs and chickens. We concluded that the SFC assay is the better method for quantitating ASC in pigs, chickens, and probably all domestic animals for which isotype-specific monoclonal antibodies are available.     

The effects of restricted feeding on adrenal cortical activity in the immature domestic fowl

1. The effect of reducing food intake to 75% of the ad libitum intake was determined from hatching to 8 weeks in young Light Sussex chickens.

2. Restricted birds were lighter throughout the experiment.

3. Relative adrenal weight tended to be greater in restricted birds but the difference decreased with time.

4. There was no depletion of adrenal cholesterol: from week 5 there was a significantly greater amount in the adrenals of restricted birds.

5. After 1 week of restriction plasma corticosterone concentration was 73% greater than in controls. It decreased progressively, falling within the normal range at 5 weeks.

6. Restricted birds were hypoglycaemic from weeks 2 to 7 and hyper‐lipacidaemic throughout. A negative correlation between plasma glucose and free fatty acids was found.     

Detection of Aphanomyces invadans and epizootic ulcerative syndrome in the Murray‐Darling drainage

Epizootic ulcerative syndrome was diagnosed, and the presence of Aphanomyces invadans confirmed, from an outbreak of clinical disease in wild‐caught bony bream (Nematalosa erebi) from the Darling River near Bourke, in New South Wales, Australia, during 2008. This confirms a significant extension of the agent beyond its historical range.     

Periodicity and gradients in dispersal ofAlternaria linicola in linseed crops

Conidia ofAlternaria linicola produced on infected linseed crops were mainly dispersed by wind. The numbers of conidia in the air above linseed crops collected by a Burkard spore sampler were greatest between 1200 h and 1300 h, when the relative humidity was lowest. Although numbers of conidia collected decreased with increasing height within and above the crop canopy, air-borneA. linicola conidia were present up to 80 cm above the crop canopy. Conidia ofA. linicola were transported by wind up to at least 40 m downwind from an artificial line inoculum source, but their numbers decreased with increasing distance from the source. In 1991, 1992, and 1993, the dispersal ofA. linicola conidia above linseed crops followed a seasonal periodicity which was influenced by weather conditions and cultural practices. The greatest numbers of conidia were collected during July, August and early September and coincided with periods favourable for sporulation and with an increase in the incidence of the disease in the senescent crop. Air-borneA. linicola conidia produced on point or line inoculum sources (naturally infected linseed stem debris) were responsible for the spread of the disease in linseed crops. In 1992 and 1993, the disease was first detected downwind from the sources, but by the end of the growing seasons, it had spread in all directions and up to 20 m and 60 m from the sources, respectively. Disease gradients were initially steep near the inoculum sources but they became flatter with time due to the secondary spread of the disease.     

Effect of apple cultivars and storage periods on the virulence of Neofabraea spp.

Bull’s eye rot is a typical quiescent postharvest apple disease in major fruit-growing areas. The susceptibility of different apple cultivars to Neofabraea spp. (N. vagabunda and N. malicorticis) was assessed, with Granny Smith showing the most resistance and Cripps Pink the most susceptibility. To assess the factors involved in conidial germination, Neofabraea spp. were grown on crude protein extracts (CPEs) collected from apple fruits at different storage periods. Fungal germ tube growth rate and pathogenic enzyme (cellulase and xylanase) activity were assessed. Results showed that CPEs collected after 2 and 4 months of storage progressively stimulated conidial germination and germ tube elongation, while a lesser effect was observed from CPEs after 1 month of storage. Xylanase proved to be the main degrading enzyme secreted by all the isolates, while cellulase was produced only by N. vagabunda isolates. Overall, the isolate ID02 was the most virulent, based on more rapid germ tube elongation and greater activity of the lytic enzymes.     

Relative importance of different types of inoculum to the establishment of Mycosphaerella graminicola in wheat crops in north‐west Europe

The contribution of wheat debris to the early stages of septoria leaf blotch epidemics was assessed in a 3‐year field experiment. First lesions were detected very early (December) in the case of an early sowing (mid‐October), showing that the first contamination could occur as soon as the seedlings emerge. The tested debris management options (chopped debris, removal of debris followed by tillage, or tillage in absence of debris) had a strong effect, although transient, on the epidemic dynamic: the more debris present on the soil surface, the more severe initial disease was. The magnitude of differences between treatments differed substantially between years. The relative production of pycnidiospores and ascospores was measured on the chopped debris. Peaks in pycnidiospore and ascospore production coincided in October–November. Both types of spores can be involved as primary inoculum in north‐west European conditions. The local amount of pycnidiospores available on debris in the field, estimated per square metre, was 1000‐fold the local ascospore production. Moreover, inoculum production was quantified on debris exposed to different environmental conditions. Autumnal conditions, characterized by moderate temperature with alternating wet and dry periods, were favourable for the production of both pycnidiospores and ascospores, as shown by the high inoculum production on debris exposed to field or outdoor conditions. By late autumn, the canopy became the most important source of pycnidiospores, and this period, characterized by the decreasing role of debris as a local source of inoculum compared to distant potential sources, can be considered as the end of the early epidemic stages.     

Application of fuzzy logic for the simulation of Plasmopara viticola using agrometeorological variables*

A mechanistic model called PLASMO was developed earlier to simulate grapevine downy mildew (Plasmopara viticola) and has been applied in several viticultural areas of Italy since 1988 by the collaboration of several research institutions of Firenze. In this study, a new simulation model based on fuzzy logic has been developed for the same structure (biological cycle of P. viticola). This approach allows classical quantitative information to be used together with qualitative information. Vague concepts can also be handled. Agrometeorological data is used, with an hourly time step, starting from budbreak to the end of the growing season. Air temperature, relative humidity, rainfall and leaf wetness are required. The simulated processes are the growth of grapevine leaf area and the main phases of the biological cycle of the pathogen: incubation, sporulation, germination, spore survival and inoculation. The main epidemiological outputs are timing of infection events and disease intensity. The performance of the model is evaluated and the mechanistic and fuzzy logic approaches are compared.