A Combination of Baiting and Different PCR Formats, Including Measurement of Real-Time Quantitative Fluorescence, For the Detection of Phytophthora fragariae in Strawberry Plants

Phytophthora fragariae, the cause of strawberry red stele disease, is a quarantine pathogen in Europe. Detecting low levels of infection requires sensitive and specific methods. In the past, Dutch and English inspection services have used bait plants to test strawberry propagation stocks destined for export. Increasingly though, PCR is being incorporated into these testing procedures in an effort to increase sensitivity and speed. Various combinations of baiting and PCR assays were compared with existing testing procedures. Water and root samples from the bait test were screened by nested PCR and the PCR amplicon was detected by several methods, including fluorescent labelled probes (TaqMan and Molecular Beacon). PCR amplification was monitored in real-time and semi-quantitative detection was possible. Because PCR reactions are sensitive to inhibitors present in extracted DNA samples, an internal control containing the primer sequences specific for P. fragariae was developed to avoid false negatives.     

Pasture and forest soil microbial communities show distinct patterns in their catabolic respiration responses at a landscape scale

Catabolic responses to specific substrates can be used to differentiate soil microbial communities. We hypothesized that the catabolic respiration responses of microbial communities from pastures would differ from those of forest soils, and that the differences would be consistent at a landscape scale, due to inherent differences in litter quality and management regimes. We analysed respiration responses to 25 different substrates of 20 pasture soils (dominated by rye grass and white clover) and 20 forest soils (indigenous forest species or the plantation species Pinus radiata) over a wide geographical range in New Zealand.Within each pasture or forest category, the catabolic responses showed a similar pattern, suggesting similarities in functional catabolic capability and microbial community Indigenous forests and pine forests microbial communities did not differ in their responses. Pasture soil communities had significantly higher relative responses to carbohydrate and amino-acid substrates and significantly lower relative response to carboxylic acid substrates, than microbial communities from forest soils. Forest soils had relatively greater responses to carboxylic acids as a group, as well as citric acid, α-ketobutyric acid, α-ketoglutaric acid, and α-ketovaleric acid, than did the pasture soils. A subset of 6 substrates was equally as effective at differentiating the microbial catabolic response of pasture soils from forest soils as the entire set of 25 substrates. The results demonstrated distinct differences in the respiration responses of the soil microbial communities of pastures and forests, but showed strong similarities within each vegetation class, despite the wide geographical spread, different soils and plant species.     

A noninvasive optical system for the measurement of xylem and phloem sap flow in woody plants of small stem size

Over the past 70 years, heat has been widely used as a tracer for estimating the flow of water in woody and herbaceous plants. However, most commercially available techniques for monitoring whole plant water use are invasive and the measurements are potentially flawed because of wounding of the xylem tissue. The study of photosynthate transport in the phloem remains in its infancy, and little information about phloem transport rates is available owing to the fragility of the vascular tissue. The aim of our study was to develop a compact, stand-alone non-invasive system allowing for direct detection of phloem and xylem sap movement. The proposed method uses a heat pulse as a tracer for sap flow. Heat is applied to the surface of the stem with a near-infrared laser source, and heat propagation is monitored externally by means of an infrared camera. Heat pulse velocities are determined from the thermometric data and related to the more useful quantity, mass flow rate. Simulation experiments on the xylem tissue of severed silver birch (Betula pendula Roth.) branch segments were performed to assess the feasibility of the proposed approach, highlight the characteristics of the technique and outline calibration strategies. Good agreement between imposed and measured flow rates was achieved leading to experimentation with live silver birch and oak (Quercus robur L.) saplings. It was demonstrated that water flow through xylem vessels can be monitored non-invasively on an intact stem with satisfactory accuracy despite simultaneous sugar transport in the phloem. In addition, it was demonstrated that the technique allows for unequivocal detection of phloem flow velocities.     

Epidemiology of parasitic gastrointestinal nematode infections of ruminants on smallholder farms in central Kenya

In order to establish the infection pattern with gastrointestinal nematodes in ruminants in the central Kenya highlands, a study was carried out in 58 smallholder farms. The study involved monthly faecal examinations from sheep, goats and cattle and pasture sampling from eight communal grazing areas. Each month, six Dorper worm-free tracer lambs were introduced and four locally grazed cross-bred sheep were purchased for parasite recovery. The mean faecal egg counts (FEC) for cattle were low throughout the study period, whereas those for sheep and goats showed a seasonal pattern with high levels of infection occurring during the two main rainy seasons, especially in March, April and October. There were significant differences in egg counts over time and among farms. Haemonchus contortus was the most prevalent nematode in the tracer lambs whereas the previously exposed locally grazed sheep had significantly lower numbers of H contortus but significantly higher numbers of Trichostrongylus species The highest levels of infection in the tracer lambs occurred in November 1995 and January, May and June 1996. Based on this study, it is now possible to explore the possibility of using strategic treatments for the control of parasitic gastroenteritis in this area of Kenya.     

Evaluation of client-specific outcome measures and activity monitoring to measure pain relief in cats with osteoarthritis

BACKGROUND: There are no validated systems for measuring pain from osteoarthritis in cats. HYPOTHESIS: Owner subjective assessments and an activity monitor (AM) can be used to detect pain in cats with osteoarthritis and to assess efficacy of treatments. ANIMALS: Thirteen cats older than 10 years old, with owner-assessed decreases in activity, painful arthritic joints, and clinically normal blood work were included and evaluated for 3 weeks. METHODS: A collar-mounted AM measured activity and a client-specific outcome measure (CSOM) questionnaire characterized the severity of impairment. Overall global quality of life was also evaluated for each treatment. In weeks 2 and 3, meloxicam (0.1 mg/kg, day 1; 0.05 mg/kg, days 2-5) or a placebo was administered in a blinded, randomized, cross-over manner to test the assessment systems. RESULTS: The cats had a median of 4 arthritic appendicular joints. Activity counts for the week when cats (complete data on activity; n=9) were administered meloxicam were significantly higher than at baseline (P = .02) but not after placebo (P = .06). Baseline activity counts were not significantly different from placebo (P = .6). The CSOM data (n=13) showed that owners considered their cats to be more active on meloxicam compared with baseline (P = .001) and placebo (P < .004), and more active on placebo than at baseline (P < .01). Global quality of life improved significantly with meloxicam (P < .042). CONCLUSIONS AND CLINICAL IMPORTANCE: Both an AM and a CSOM system can detect behavior associated with pain relief in cats that are arthritic. Objective activity data might allow subjective assessment systems to be validated for use in clinical studies.     

Assessing the performance against houseflies of indoor aerosol space sprays. Part I. Factors examined during tests with houseflies in free flight

The development of a mathematical model which, by quantifying the interaction of a population of houseflies with an ageing spray cloud, is capable of predicting the knockdown and kill performance of insecticidal aerosol formulations shown against houseflies, Musca domestica L., will be presented in a series of four separate papers. In this first paper, measurements were made of insecticide collected by houseflies during free flight using a standard room-spray technique. With pressure-packed aerosol formulations where the solvent is the principal change, it was shown that both knockdown and mortality responses of the insects were directly related to dose collected. The process of droplet collection was most significantly influenced by their size and availability, dependent upon the time after spraying, the position of insect release and the activity of individual adult M. domestica. The knockdown or mortality response of the insect to given amounts of insecticide was largely unaffected by the remaining solvent, although, with mineral oil, some inhibition was noted at low mortality levels. The amount of insecticide accumulated by houseflies in their initial flight was of prime importance in determining the overall effect. Flies remaining not knocked down at the end of the exposure period were shown to be so because they had accumulated less toxicant and not necessarily because they were more tolerant of the insecticide. The establishment of a relationship between the number of flies in flight and time after release has provided a key function for incorporation into the model.