Relative contribution of edge and interior zones to patch size effect on species richness: An example for woody plants

In order to understand the capacity of habitats to conserve species, many authors have searched for a species–area relationship (SAR) to evaluate the effect of patch size on species richness in habitat fragments. However, a range of different processes may underlie or obscure this relationship. For woody plant species in forest fragments, as for other taxa, considering forest edges separately in the investigation of SAR is particularly relevant. The objective of our study was to evaluate edge influence on SAR in a fragmented forest landscape in south-western France. To achieve this objective, we considered SAR in two separate spatial portions of the forest fragment: the edge and the forest interior. We also considered SAR for different ecological groups of species based on their shade tolerance and their mode of dispersal, as species can respond differently to habitat reduction depending on their ecological characteristics. In 40 woodlands in an agricultural landscape, we observed the presence/absence of all woody species along parallel walking transects 20-m wide to inventory the total number of species in each woodlot. Vegetation surveys resulted in the identification of 53 woody species, with a total of 23 trees and 30 shrubs, and a number of species per woodlot ranging from 18 to 44. The species richness found in a given woodlot was significantly correlated with its area. When considering the edge and the interior parts of the woodlot separately we found a steeper increase in species richness with area for the part we considered as the edge, compared with the increase found in the interior part of the forest. Our results confirm the contribution of forest edge to forest fragment SAR. Results also underlined the importance of two additional processes that may contribute to SAR: a possible extinction debt of shade-tolerant species in forest edge zones and colonisation by light-demanding species in forest interior zones probably due to disturbances. This approach underlines the importance of taking the edge effect into account when studying the influence of patch size on species richness and the dynamic of species richness pattern.     

Effects of animal position and number of repeated measurements on selected two-dimensional and M-mode echocardiographic variables in healthy dogs

OBJECTIVE: To evaluate the effects of positioning and number of repeated measurements on intra- and interobserver variability of echocardiographic measurements in dogs. DESIGN: Prospective study. ANIMALS: 4 healthy dogs. PROCEDURE: Each observer performed 24 examinations, separately assessing each dog 6 nonconsecutive times (3 times with the dog in lateral recumbency and 3 with the dog in a standing position). Variables evaluated included M-mode measurements of left ventricular end-diastolic and left ventricular end-systolic diameters, left ventricular free-wall thickness in diastole and systole, interventricular septal thickness in diastole and systole, left ventricular shortening fraction, and 2-dimensional measurements of the left atrial diameter-to-aortic diameter ratio. RESULTS: All coefficients of variation (range, 3.4% to 26.6%) were similar between operators and positions and were < 15% for 27 of 32 values. For both operators, repeatability of the measurements was better for left ventricular end-systolic diameter, left ventricular free-wall thickness in diastole, left ventricular free-wall thickness in systole, and the left atrial diameter-to-aortic diameter in the standing position, and similar for both positions for shortening fraction and left ventricular end-diastolic diameter. No effect of cardiac cycle was observed. CONCLUSIONS AND CLINICAL RELEVANCE: Within-day variability of conventional echocardiography performed with the dog in the standing position was at least as good as that obtained with the dog in lateral recumbency for most measured variables. Single measurements of each variable may be sufficient for trained observers examining dogs that do not have an arrhythmia. The standing position should be used, particularly for stressed or dyspneic dogs.     

Spatial heterogeneity of the risk of BSE in France following the ban of meat and bone meal in cattle feed

In France, meat-and-bone meal (MBM) has been prohibited for cattle feeding since 1990, but bovine spongiform encephalopathy (BSE) cases, called 'NAIF', appeared in animals born after this feed ban. Furthermore, in 1996 a new measure was taken: removal of cadavers and specified risk materials (SRM) from the processing of MBM dedicated to animal feed. Nevertheless, BSE cases (called 'super-NAIF') appeared in cattle born after this measure was in force. We analysed the spatial distribution of 445 'NAIF' and 58 'super-NAIF' cases detected in France from July 1, 2001 to July 31, 2003. The detection of BSE was based both on the mandatory reporting system (MRS) and the systematic test screening of cattle at the abattoir and at the fallen-animal plant with rapid tests. The background population was based on the adult-cow census. The disease mapping of the BSE risk was based on the standardised incidence ratio (stochastic Poisson process). A spatial component, which takes into account the spatial dependence between the geographical units by a notion of adjacency was used to eliminate the over-dispersion in the risk assessment. The geographical units were defined by hexagons with a side of 23km (France had 1264 hexagons). The parameters were estimated by a Metropolis Gibbs sampling algorithm using the Markov-chain Monte Carlo methods. The BSE cases were not randomly distributed. Furthermore, the areas at risk for the 'super-NAIF' matched part of the areas at risk for the 'NAIF' cases-which suggests that it might be a common source of contamination.     

A spatio-temporal analysis of BSE cases born before and after the reinforced feed ban in France

A spatio-temporal analysis was carried out to see how the risk distribution of bovine spongiform encephalopathy (BSE) in France changed depending on the period of birth. The data concerned the 539 BSE cases born in France after the ban (BAB) of meat and bone meal (MBM) in 1990 and detected between July 1, 2001 and December 31, 2003, when the surveillance of BSE was comprehensive. Seventy-two of these cases were born after the reinforced (second) ban (BASB) in 1996, which involved the removal of BSE-risk materials and cadavers from the processing of MBM. The Ederer-Myers-Mantel (EMM) time and space cluster test was applied, after classifying the cases by trimester and region of birth, BAB or BASB status, and dairy or beef status. Then disease mapping was performed for four successive birth periods, three for the BAB cases (January 1991 through June 1994, July 1994 through June 1995, July 1995 through June 1996), and one for the BASB (July 1996 through October 1998). It was elaborated with the Bayesian graphical modelling methods and based on a Poisson distribution with spatial smoothing. The parameters were estimated by a Markov Chain Monte Carlo (MCMC) simulation method. The main finding was that the areas with the highest risk of BSE changed largely from one birth period to another; from the west, it reached the east of France for birth cohort 1994-1995 and the southwest for birth cohort 1995-1996. The EMM test identified a peak risk in this region both for dairy and beef cattle in the fall 1995. The spatial distribution of the risk for the BASB cases matched the spatial pattern of risk for the preceding BAB birth cohort quite well; this was in favour of a common origin of the infection of the BAB and BASB cases, despite the complementary control measures.     

Development of an enzyme-linked immunosorbent assay for specific equine neutrophil myeloperoxidase measurement in blood.

Equine inflammatory disease is accompanied by a neutrophil activation resulting in the release of granulocytic enzyme myeloperoxidase (MPO). To measure MPO in horse plasma as marker of neutrophil activation, the authors purified equine neutrophil MPO and developed a specific enzyme immunoassay using 2 specific polyclonal antibodies obtained from rabbit (primary antibody) and guinea pig (secondary antibody). The sandwich complex "primary antibody-MPO-secondary antibody" was detected using a goat anti-guinea pig immunoglobulin antibody conjugated to alkaline phosphatase. The enzyme-linked immunosorbent assay (ELISA) showed good precision and accuracy, with intra- and interassay coefficients of variation below 10% for MPO concentrations ranging from 0.78 to 50 ng/ml. A stable plasma MPO value, unaffected by time elapsed between blood collection and centrifugation, was obtained with plasma from EDTA anticoagulated blood. The mean MPO value measured in 38 healthy horses was 181.80 +/- 64.74 ng/ml. In 20 horses suffering from obstruction of the large or small intestine, MPO concentrations measured at the time of arrival at the intensive care unit were significantly higher than mean normal value, ranging from 477.88 to 2,748.13 ng/ml. Work is in progress to apply this MPO ELISA technique to other biological fluids and other equine diseases.     

Assessment of repeatability, reproducibility, and effect of anesthesia on determination of radial and longitudinal left ventricular velocities via tissue Doppler imaging in dogs

OBJECTIVE: To determine left ventricular free wall (LVFW) motions and assess their intra- and interday variability via tissue Doppler imaging (TDI) in healthy awake and anesthetized dogs. ANIMALS: 6 healthy adult Beagles. PROCEDURE: n the first part of the study, 72 TDI examinations (36 radial and 36 longitudinal) were performed by the same observer on 4 days during a 2-week period in all dogs. In the second part, 3 dogs were anesthetized with isoflurane and vecuronium. Two measurements of each TDI parameter were made on 2 consecutive cardiac cycles when ventilation was transiently stopped. The TDI parameters included maximal systolic, early, and late diastolic LVFW velocities. RESULTS: The LVFW velocities were significantly higher in the endocardial than in the epicardial layers and also significantly higher in the basal than in the mid-segments in systole, late diastole, and early diastole. The intraday coefficients of variation (CVs) for systole were 16.4% and 22%, and the interday CV values were 11.2% and 16.4% in the endocardial and epicardial layers, respectively. Isoflurane anesthesia significantly improved the intraday CV but induced a decrease in LVFW velocities, except late diastolic in endocardial layers and early diastolic in epicardial layers. CONCLUSIONS AND CLINICAL RELEVANCE: Left ventricular motion can be adequately quantified in dogs and can provide new noninvasive indices of myocardial function. General anesthesia improved repeatability of the procedure but cannot be recommended because it induces a decrease in myocardial velocities.     

Estimating the trend of the French BSE epidemic over six birth cohorts through the analysis of abattoir screening in 2001 and 2002

A bovine spongiform encephalopathy (BSE) testing programme at the abattoir started in 2001 in France. A total of 5 281 293 bovines were tested in 2001 and 2002; 87 were found positive in 2001--37 per million (95% CI 30-46)--, whereas only 71 in 2002--24 per million (95% CI 19-30). Logistic regression models were run to compare the prevalence of BSE on successive birth cohorts, using a pair-wise method of controlling for age at testing; the prevalence on the first one, determined on animals slaughtered in 2001, was compared to the prevalence on the following one determined on animals slaughtered in 2002. Five models were performed in order to compare the birth cohorts preceding and following the months of June 1993 (i.e. July 92-June 93 birth cohort compared to July 93-June 94 birth cohort) (8.5 years old cattle), June 1994 (7.5 years old cattle), June 1995 (6.5 years old cattle), June 1996 (5.5 years old cattle) and June 1997 (4.5 years old cattle). The models were adjusted for the production type of cattle and the test used. The results showed a significant increase (OR = 2.31, 95% CI 1.08-4.9) of the BSE prevalence between the July 93-June 94 and July 94-June 95 cohorts, and then a significant decrease over the next two birth cohorts; the July 95-June 96 birth cohort was significantly less affected than the July 94-June 95 one (OR = 0.46, 95% CI 0.27-0.78), and the July 96-June 97 birth cohort was significantly less affected than the July 95-June 96 one (OR = 0.17, 95% CI 0.07-0.37). The increase in BSE prevalence between the July 93-June 94 and July 94-June 95 cohorts was in agreement with modelling studies, but needs to be confronted to the data on fallen stock at the national level. The decrease in BSE prevalence on the birth cohorts born after June 1995 was in agreement with the findings on the fallen stock in the western part of France and matches the implementation of the removal of specified risk materials (SRM) and dead animals from the processing of meat and bone meal (MBM) since June 1996.     

Temporal and spatial patterns of the clinical surveillance of BSE in France,analysed from January 1991 to May 2002 through a vigilance index

Between 1991 and mid 2000, the surveillance of Bovine Spongiform Encephalopathy (BSE) in France was based solely on clinical surveillance through a Mandatory Reporting System. Since 2000, the implementation of active surveillance programmes using rapid tests, as a complementary tool targeted at dead and slaughtered cattle has shown that part of the BSE cases were not detected with the clinical surveillance. In order to obtain a better knowledge of the strength of the clinical surveillance, we analysed a vigilance index defined as the ratio of negative clinical suspicions to the cattle population in the region and period of interest. The temporal analysis of the vigilance index showed that it did not vary much between 1991 and 1999, increased sharply since 2000, and then decreased partly in 2001. The geographical analysis of the variations of the vigilance index was performed at the department level by comparing the observed number of negative clinical suspicions per department to the expected number, computed on the basis of the national average index and standardised on the production type of the cattle - dairy versus beef suckling cattle. As assumed, the data followed a Poisson distribution. We observed a high geographical variation of the vigilance index: ten departments out of 91 presented a significantly higher vigilance index than the national one, and four a significantly lower vigilance index. The vigilance index showed that the clinical surveillance was heterogeneous during the past twelve years, both in time and geographic location, in a range of one to ten. So the apparent trend in the BSE epidemic during this period as well as the differences in the spatial incidence of BSE have to be analysed with caution.     

Equine trypsin: purification and development of a radio-immunoassay

Shock is accompanied by generalised splanchnic hypoperfusion, and splanchnic organs like the pancreas can be damaged, as shown in animal experimental models and in humans, by the presence of high plasma concentrations of trypsin and other pancreatic enzymes. In order to design a radioimmunoassay technique (RIA) for the measurement of equine trypsin-like immunoreactivity (TLI) in biological fluids, trypsin was purified (with purity > or = 96%) from the equine pancreas by extraction in an acid medium, ammonium sulfate precipitations, gel filtration chromatography and, after activation of trypsinogen into trypsin, affinity chromatography. Gel polyacrylamide electrophoresis showed a monomeric enzyme with a molecular weight of 27 kDa. The purified equine trypsin served for the immunisation of rabbits in order to obtain a specific antiserum, and the labelled antigen was prepared by iodination of equine trypsin with 125I. The RIA was based on the binding of the antigen to the antibody followed by the separation of the antigen-antibody complex by immunoprecipitation in the presence of sheep anti-rabbit gammaglobulins and the assay of the radioactivity in the precipitate. The RIA showed good sensitivity, specificity, precision, accuracy and reproducibility. The reference mean value of TLI in the plasma of healthy horses (n = 20) was 30.01+/-6.84 ng/mL (upper confidence limit 50.52 ng/mL; p < 0.01). Three horses with non strangulating intestinal obstruction without shock showed TLI values within normal limits whereas 5 of 7 horses with strangulation obstruction showed TLI levels above the upper confidence limit. Further studies using the RIA and the enzymatic assay should be performed in order to confirm the role of the pancreas in equine intestinal obstruction.