Microscopic analysis of the effect of azoxystrobin treatments on Mycosphaerella graminicola infection using green fluorescent protein (GFP)‐expressing transformants

Green fluorescent protein (GFP)‐expressing transformants were used to investigate the effects of strobilurin fungicide azoxystrobin on Mycosphaerella graminicola infection. Azoxystrobin treatments (125 or 250 g AI ha^?1) were applied at various stages of the infection process under controlled conditions. GFP transformants showed conserved in vitro sensitivity to azoxystrobin and pathogenicity. Azoxystrobin controlled over 90% of M graminicola infections when applied before or during penetration of the pathogen (15% of the incubation phase). Azoxystrobin also impaired the growth of intercellular hyphae in M graminicola post‐penetration infection stages when applied at up to 50% of the incubation phase. Incubating infections observed in treated leaves were viable, but their growth was impaired and they did not induce necrosis under controlled conditions. Reduction by half of azoxystrobin dosage had little or no effect on azoxystrobin efficiency in controlling M graminicola. The contribution of post‐penetration fungistatic effect to azoxystrobin curative properties toward M graminicola in a field situation is discussed. © 2001 Society of Chemical Industry     

The transmission by nematodes of tobraviruses is not determined exclusively by the virus coat protein

The coat protein gene of the nematode non-transmissible, SP5 isolate of pea early-browning tobravius was replaced with that of the highly nematode transmissible, PPK20 isolate of tobacco rattle tobravirus. Plants were infected with the recombinant virus when mechanically inoculated and the virus invaded the plants systemically. However, although the PPK20 isolate of TRV was transmitted by nematodes from these plants, the recombinant virus was not transmitted. Therefore, the virus coat protein is not the exclusive determinant of nematode transmission.     

Purification and analyses of the specificity of two putative diagnostic antigens for larval cyathostomin infection in horses

Cyathostomins are important equine gastrointestinal parasites. Mass emergence of mucosal stage larvae causes a potentially fatal colitis. Mucosal stages are undetectable non-invasively. An assay that would estimate mucosal larval stage infection would greatly assist in treatment, control and prognosis. Previously, we identified two putative diagnostic antigens (20 and 25 kDa) in somatic larval preparations. Here, we describe their purification and antigen-specific IgG(T) responses to them. Western blots confirmed the purity of the antigens and showed that epitopes in the 20 kDa complex were specific to larval cyathostomins. No cross-reactive antigens appeared to be present in Parascaris equorum or Strongyloides westeri species. Low levels of cross-reactivity were observed in Strongylus edentatus and Strongylus vulgaris species. Use of purified antigens greatly reduced background binding in equine sera. These results indicate that both antigen complexes may be of use in a diagnostic assay.     

The effects of stock crate design and stocking density on environmental conditions for lambs on road transport vehicles

AIM: The aim of this study was to evaluate the potential of variations in stock crate design (especially ventilation) and stocking density on road transport vehicles for reducing the risk of environmental stress for lambs during long-haul transport in hot weather. METHODS: In Experiment 1, lambs were transported on vehicles fitted with either a newer-design aluminium crate comprising a three-deck truck and four-deck trailer, or an older-design, more open, steel crate of a three-deck truck and three-deck trailer. In Experiment 2, lambs were transported on newer-design vehicles at either standard stocking density (0.20 m2 per 35 kg lamb) or at a 20% lower density (0.26 m2 per 35 kg lamb). In each experiment, each journey was replicated twice and consisted of travel periods and stationary periods designed to emulate conditions associated with an inter-island ferry crossing. Air ammonia concentrations and temperature and humidity were monitored within six pens on each vehicle, and the temperature-humidity index (THI) was calculated. RESULTS: Ammonia concentrations were variable and generally 50 ppm, and did not vary significantly with treatment. The THI increased when the vehicles were stationary, especially under conditions designed to emulate an enclosed ferry deck. The ambient temperature during Experiment 1 was moderate (up to 21 degrees C), and THI was slightly lower in the older-design crate. High ambient temperatures (up to 33 degrees C) were present during Experiment 2, and THI was significantly lower at the lower stock density. During a 3-h stationary period, the peak THI at standard stocking density was 91.0, compared with 84.9 for the low density treatment (p<0.001). CONCLUSIONS: For standard road transport vehicles used for long-haul transport of lambs, lowered loading density may be of considerable benefit in alleviating conditions that increase the risk of lamb deaths during inter-island transport on hot days.     

Antigen-specific IgG(T) responses in natural and experimental cyathostominae infection in horses

Equine clinical larval cyathostominosis is caused by simultaneous mass emergence of previously inhibited larvae from the mucosa of the colon. Clinical signs include diarrhoea, colic, weight loss and malaise, and in up to 50% of cases, the disease results in death. Cyathostominae spend a large part of their life cycle as larval stages in the intestinal mucosa. Definitive diagnosis is difficult due to the lack of diagnostic methods for pre-patent infection. In the present study, the enzyme-linked immunosorbent assay (ELISA) was used to investigate isotype responses to larval cyathostominae somatic antigen. Measurement of anti-larval IgG(T) responses appeared to have the most immunodiagnostic potential. An increase in IgG(T) response was detected to crude larval antigen by 5 weeks post-infection (PI) in individual infected ponies. Subsequently, IgG(T) responses to larval and adult somatic extracts were examined by Western blotting using sera from experimentally-infected horses and helminth-naive animals (n=6). Two antigen complexes, designated A and B, in larval somatic antigen were recognised specifically by the infected animals by 7 weeks PI. Sera taken from 23 endemically-infected animals, whose cyathostominae burdens had been enumerated, were also used to identify putative diagnostic antigens. Eighteen horses had positive mucosal worm burdens (range 723-3,595,725) and all but two of these animals had serum IgG(T) antibody specific to either complex. Moreover, IgG(T) responses specific to antigen complexes A and B were absent in all five parasite negative horses that were tested. Serum IgG(T) responses to either of the two complexes were identified in five clinical cases tested. IgG(T) responses to adult antigen somatic extracts were more heterogeneous, with no clear pattern between experimentally-infected ponies and helminth-free controls. The results indicate that increases in serum IgG(T) to mucosal larvae occur in the pre-patent period and that two antigenic complexes within somatic preparations of these stages have immunodiagnostic potential.     

Quantitative PCR monitoring of the effect of azoxystrobin treatments on Mycosphaerella graminicola epidemics in the field

Quantitative PCR and visual monitoring of Mycosphaerella graminicola epidemics were performed to investigate the effect of curative and preventative applications of azoxystrobin in wheat field crops. A non-systemic protectant and a systemic curative fungicide, chlorothalonil and epoxiconazole, respectively, were used as references. PCR diagnosis detected leaf infection by M graminicola 3 weeks before symptom appearance, thereby allowing a clear distinction between curative and preventative treatments. When applied 1 week after the beginning of infection, azoxystrobin curative activity was intermediate between chlorothalonil (low effect) and epoxiconazole. When applied preventatively, none of the fungicides completely prevented leaf infection. There was some indication that azoxystrobin preventative treatments may delay fungal DNA increase more than epoxiconazole at the beginning of leaf infection. Both curative and preventative treatments increased the time lapse between the earliest PCR detection and the measurement of a 10% necrotic leaf area. Azoxystrobin only slightly decreased the speed of necrotic area increase compared with epoxiconazole. Hence, azoxystrobin activity toward M graminicola mainly resides in lengthening the time lapse between the earliest PCR detection and the measurement of a 10% necrotic leaf area. Information generated in this way is useful for optimal positioning of azoxystrobin treatments on M graminicola.     

Agent-based land-use models: a review of applications

Agent-based modelling is an approach that has been receiving attention by the land use modelling community in recent years, mainly because it offers a way of incorporating the influence of human decision-making on land use in a mechanistic, formal, and spatially explicit way, taking into account social interaction, adaptation, and decision-making at different levels. Specific advantages of agent-based models include their ability to model individual decision-making entities and their interactions, to incorporate social processes and non-monetary influences on decision-making, and to dynamically link social and environmental processes. A number of such models are now beginning to appear—it is timely, therefore, to review the uses to which agent-based land use models have been put so far, and to discuss some of the relevant lessons learnt, also drawing on those from other areas of simulation modelling, in relation to future applications. In this paper, we review applications of agent-based land use models under the headings of (a) policy analysis and planning, (b) participatory modelling, (c) explaining spatial patterns of land use or settlement, (d) testing social science concepts and (e) explaining land use functions. The greatest use of such models so far has been by the research community as tools for organising knowledge from empirical studies, and for exploring theoretical aspects of particular systems. However, there is a need to demonstrate that such models are able to solve problems in the real world better than traditional modelling approaches. It is concluded that in terms of decision support, agent-based land-use models are probably more useful as research tools to develop an underlying knowledge base which can then be developed together with end-users into simple rules-of-thumb, rather than as operational decision support tools. This paper arises from research conducted as part of the UK Research Councils’ RELU Programme (award number RES-224-25-0102). RELU is funded jointly by the Economic and Social Research Council, the Biotechnology and Biological Sciences Research Council and the Natural Environment Research Council, with additional funding from the Department for Environment, Food and Rural Affairs and the Scottish Executive Environment and Rural Affairs Department.