In vitro viability of cryopreserved equine embryos following different freezing protocols.

The main objective of this study was to evaluate two freezing protocols and the effect of agar embedding on survival of day 6.5 equine embryos. A total of 133 embryos were used, in one group (n = 51), embryos were first embedded in agar before the freezing protocol was started. A freezing protocol to -30 degrees C or -33 degrees C was used before plunging embryos into liquid nitrogen (LN2). The embryos were thawed in water at 37 degrees C, evaluated and placed in culture. After 24 h culture, the embryos were evaluated for their morphology and development. No differences were observed between embryos plunged at -30 degrees or at -33 degrees C in LN2. The analysis of the morphology and development after thawing showed that the diameter and developmental stage at freezing correlated with embryo survival. Morula and early blastocyst stages of development were associated with better quality after freezing and thawing and had a better potential to survive after in vitro culture (p < 0.05) compared to more advanced stages. The agar failed to protect embryos from zona pellucida damage, but a tendency to prevent rupture was observed in larger embedded embryos.     

Physiopathologie et thérapeutique de l'utérus en période puerpérale chez la vache laitière: revue

Accurate diagnosis and effective therapeutics of uterine pathologies in cattle represent some of the daily preoccupations for the veterinarians involved in herd health medicine. A rational approach requires a competence based on fundamental and clinical knowledge.

The physiological aspects of the uterine involution in cattle are reviewed with emphasis about the in utero bacteriological dynamic and the endocrinological evolution during this period. The pathogenesis of uterine pathologies is discussed following new research results and new knowledge on the mechanisms of uterine defense. The value and limitations of the clinical diagnosis is briefly reviewed. The therapeutics of uterine pathologies is debated in relation with the pathophysiology of the uterine involution and with the rational of the traditional and new concepts of the treatment of the postpartum uterine pathologies.

     

Effect of the Pietrain breed used as terminal boar on Post-weaning Multisystemic Wasting Syndrome (PMWS) in the offspring in four PMWS-affected farms

The aim of this study was to determine the potential risk factors for PMWS at the individual pig level and assess the effect of the Pietrain paternal genetic background of the animals in a cohort study. The survey was set up in four PMWS-affected farms with 2 repetitions (batches) per farm. A representative sample of 60 pigs per batch, stratified according to the paternal genetic background (Pietrain: yes vs. no), was randomly selected after farrowing. The representative cohort was divided into 8 batches and the pigs were individually monitored from birth to slaughter. Survival analysis was used to determine the factors related to the time to PMWS. The litter-cluster effect was taken into account using the marginal Cox model (robust estimation of the covariance matrix) and the gamma shared frailty model which were compared.No protective effect of the Pietrain breed on the time to PMWS and the proportion of affected pigs in the offspring was found in this study. Piglets showing low circovirus type 2 (PCV2) titres at 7 weeks-old with no subsequent seroconversion and piglets from PCV2 negative sows were most likely to be affected by PMWS (HR=7.0 and 2.8, respectively). Active infection of the pregnant dams with parvovirus was related to an increased risk of PMWS in the offspring (HR=2.3). Neck injuries due to poorly performed injections in the dams were associated with an increased risk of PMWS with the marginal model (HR=2.1). Oxytocin injection (dams) during farrowing was protective against PMWS in the offspring (HR=0.6).     

Some determinants of the spatio-temporal fire cycle in a mediterranean landscape (Corsica,France)

Based on recent needs to accurately understand fire regimes and post-fire vegetation resilience at a supra-level for carbon cycle studies, this article focusses on the coupled history of fire and vegetation pattern for 40 years on a fire-prone area in central Corsica (France). This area has been submitted since the beginning of the 20^th century to land abandonment and the remaining land management has been largely controlled by frequent fires. Our objectives were to rebuild vegetation and fire maps in order to determine the factors which have driven the spatial and temporal distribution of fires on the area, what were the feed backs on the vegetation dynamics, and the long-term consequences of this inter-relationship. The results show a stable but high frequency of small fires, coupled with forest expansion over the study period. The results particularly illustrate the spatial distribution of fires according to topography and vegetation, leading to a strong contrast between areas never burnt and areas which have been burnt up to 7 times. Fires, when occuring, affect on average 9 to 12% of the S, SE and SW facing slopes (compared to only 2 to 5% for the N facing slopes), spread recurrently over ridge tops, affect all the vegetation types but reburn preferentially shrublands and grasslands. As these fire-proning parameters have also been shown to decrease the regeneration capacity of forests, this study highlights the needs in spatial studies (both in terms of fire spread and vegetation dynamic) to accurately apprehend vegetation dynamic and functionning in fire-prone areas.This revised version was published online in May 2005 with corrections to the Cover Date.     

New screening test to predict the potential impact of ivermectin-contaminated cattle dung on dung beetles

According to European Union recommendations, a test method has been developed to evaluate the effects of veterinary pharmaceuticals on dung feeding insects. This test method was evaluated with the dung beetle Aphodius constans by using fecal residues of ivermectin after a pour-on administration. Dung of different age (and thus containing different concentrations of ivermectin) as well as mixtures of highly-contaminated spiked dung with untreated control dung were studied in five test runs in two laboratories. The concentration of ivermectin (active substance; a.s.) in the dung samples was verified analytically. The main test endpoint was the survival of first instar larvae. The LC50 using dung directly obtained from treated cattle ranged from 470 to 692 microg a.s. kg(-1) dung (dry weight; d.w.) and 67 to 97 microg a.s. kg(-1) dung (fresh weight; f.w.). Using mixtures, the outcome of two tests was almost identical: 770 to 781 microg a.s. kg(-1) dung (d.w.); 109 to 132 microg a.s. kg(-1) dung (f.w.). In comparison to the LC50 values obtained when ivermectin was spiked in control dung at several concentrations (LC50 880-985 microg a.s. kg(-1) dung (d.w.)), the LC50 values were again very similar. Three conclusions can be drawn from these results. The proposed test method seems to be robust and allows for the initiation of an international validation process (including ringtesting). Because of only small differences found in tests in which the test substance was spiked into control dung and those in which dung from treated cattle was applied, the use of a standard test method is proposed. The effects of ivermectin on ecologically relevant dung beetles obtained in a standardised test method reflect the results from field studies and are in the range of environmentally relevant concentrations.     

Rapid seedlings regeneration from seeds and vegetative propagation with sucker and rhizome of Eremospatha macrocarpa (Mann & Wendl.) Wendl and Laccosperma secundiflorum (P. Beauv.) Kuntze

To develop efficient seedling production methods for Laccosperma secundiflorum and Eremospatha macrocarpa, a study was conducted to examine regeneration using offsets combined with several physical and chemical treatments of seeds. Offsets categorized into small, medium and large diameters, were planted in three conditions: shaded and open nursery, and greenhouse. We tested sucker from E. macrocarpa, and sucker and rhizome from L. secundiflorum. For both species, high viability percentage (ranging from 55% to 100%) were observed for small and medium suckers planted in shaded nursery and greenhouse, against less than 49% for sucker planted in open nursery. The mean seedling emergence times were estimated to 84, 77 and 75 days after planting (DAP) for small, medium and large sucker of L. secundiflorum, respectively under open nursery condition, and 76, 75, 95 DAP for small, medium and large suckers of the same species, respectively in shaded condition. Greenhouse has a significant positive effect on E. macrocarpa seedlings emergence time. For this species, the mean seedling emergence times were estimated to 43 DAP for small sucker and 76, 93 DAP for medium and large suckers. No seedling was obtained from rhizome planted in all the growing conditions tested. Concerning seed dormancy breaking, germination percentages and rates were determined for 13 treatments. The best treatments were pre-soaking unscarified seeds for 4 days in 1.01 g l⁻¹ and 0.10 g l⁻¹ KNO₃, with 79% and 68% of germination, respectively and in 3.46 × 10⁻³ g l⁻¹ GA₃ for 68% of germination. These methods are suggested to improve germination of L. secundiflorum seeds. Successful and recommended methods for E. macrocarpa are pre-soaking scarified seeds in 3.46 × 10⁻³ g l⁻¹ and 3.46 × 10⁻⁴ g l⁻¹ GA₃, 96% and 94% of germination, respectively. Dormancy, probably a combination of mechanical and chemical dormancy, is present in the two species.     

Evolution of bacterial resistance to certain antibacterial agents in horses in a veterinary hospital

Evolution of bacterial resistance to certain antibacterial agents in horses in a veterinary hospital. A total of 255 antibiograms, conducted according to the Kirby-Bauer method from bacterial isolates collected from horses at the H?pital Vétérinaire d'Enseignement at the Université de Montréal between 1996 and 1998 were compared with the results obtained about 10 years ago with corresponding bacterial species. A significant increase in the percentage of strains resistant to trimethoprim-sulfamethoxazole (TMP-SXT), penicilline, tetracycline, and to gentamicin was observed. The percentages of isolates of Actinobacillus spp. and coagulase-positive staphylococci resistant to TMP-SXT increased from 0% to 17% and from 0% to 33%, respectively, (P < 0.05). The percentage of isolates of Actinobacillus spp resistant to penicilline also increased from 33% to 73% (P < 0.05). Regular monitoring of the evolution of bacterial resistance to antibacterial agents is indispensable in a hospital structure. The adoption of "good antibiotherapy practices" is essential to guarantee a decrease in the risks of selection and dissemination of a resistant bacterial strain.     

Prevalence of enteropathogenic Escherichia coli in naturally occurring cases of poult enteritis-mortality syndrome

Enteropathogenic Escherichia coli (EPEC) previously were identified in poult enteritis-mortality syndrome (PEMS)-affected turkeys and associated as a cause of this disease. In the present study, the prevalence of EPEC in PEMS-affected turkeys was examined retrospectively with archived tissues and intestinal contents collected from 12 PEMS-affected turkey flocks in 1998. Formalin-fixed intestinal tissues were examined by light and electron microscopy for attaching and effacing (AE) lesions characteristic of EPEC, and frozen (-75 C) intestinal contents were examined for presence of EPEC. Escherichia coli isolates were characterized on the basis of epithelial cell attachment, fluorescent actin staining (FAS) test, and presence of E. coli attaching/effacing (EAE), shigalike toxin (SLT) type I, SLT II, and bundle-forming pilus (BFP) genes by polymerase chain reaction procedures. EPEC isolates were examined for pathogenicity and ability to induce AE lesions in experimentally inoculated young turkeys. AE lesions were identified by light microscopy in Giemsa-stained intestines from 7 of 12 PEMS-affected turkey flocks. Lesions consisted of bacterial microcolonies attached to epithelial surfaces with epithelial degeneration at sites of attachment and inflammatory infiltration of the lamina propria. Electron microscopy confirmed the identity of AE lesions in six of seven flocks determined to have AE lesions by light microscopy. EPEC were identified in 4 of 12 flocks on the basis of the presence of EAE genes a nd absence of SLT I and SLT II genes; all isolates lacked BFP genes. EPEC isolates produced AE lesions and variable mortality in turkeys coinfected with turkey coronavirus. In total, EPEC were associated with 10 of 12 (83%) naturally occurring PEMS cases on the basis of identification of AE lesions and/or EPEC isolates. These findings provide additional evidence suggesting a possible role for EPEC in the pathogenesis of PEMS.     

Protective Effects Against Erwinia amylovora Induced by hrp Mutants in the Whole Plant are Only Partially Mimicked in Cultivated Cells

A virulent strain of Erwinia amylovora, the causal agent of fire blight of Maloideae, and two of its non-virulent hrp mutants (a secretory and a regulatory mutant) were inoculated into apple cell suspensions either alone or in mixed inoculations. In single inoculations, death of 4- to 5-day-old apple cells occurred only when the concentration of the virulent strain of E. amylovora reached a threshold inoculum concentration of 10⁴CFUml^–1, while high concentrations of the hrp mutants were unable to kill apple cells. When mixed inoculated with the virulent parental strain, both hrp mutants protected apple cells from death caused by the virulent strain. The protective effect was associated with a decrease in the population level of the virulent strain and it was dependent on the non-virulent to virulent inoculum concentration suggesting a competition between the non-virulent mutant and the virulent strain. However, no differential protective ability between the two types of mutants could be noticed, contrary to previous results obtained with apple seedlings or apple flowers in which the regulatory mutant was significantly more effective than the secretory mutant. In conclusion, inoculation of apple cell cultures with E. amylovora does not seem to be a model suitable for investigating mechanisms leading to protection.