Epidemiological investigations in regard to porcine reproductive and respiratory syndrome (PRRS) in Quebec, Canada. Part 2: prevalence and risk factors in breeding sites

Porcine reproductive and respiratory syndrome virus (PRRSV) is a major threat for swine industry and understanding factors involved in its epidemiology is undoubtedly essential for disease control. As a part of a larger project, a cross-sectional study was performed on breeding sites in a moderate density area of swine production in Quebec to estimate the prevalence of PRRSV infected sites and to evaluate if characteristics of sites and biosecurity practices, either as specific measures or as a global score, were associated with PRRSV status. A questionnaire and diagnostic procedures were performed on 54 breeding sites between September 2006 and August 2008. A biosecurity score that had been previously computed using two-step clustering procedure was used, classifying breeding sites into two biosecurity patterns (high vs. low) according to 21 specific biosecurity measures. The apparent prevalence of PRRSV infected sites was 74.0% (95% CI, 60.3-85.0). Univariable and multivariable logistic regression models with robust standard errors adjusting for potential clustering of sites due to same ownership were computed. In a first multivariable model evaluating characteristics of sites and specific biosecurity variables, four main effects were significantly associated (P<0.05) with PRRSV positive status: large pig inventory (OR: 10.7), proximity to closest pig site (OR: 7.3), absence of shower (OR: 8.7) and free access to the main entrance of the site by the rendering truck (OR: 7.0). In a second multivariable model including a global biosecurity score as a surrogate for a specific pattern of biosecurity measures, this score was not retained in the final model. The adjusted population attributable fractions were 16% for the proximity to closest pig site variable, 27% for the absence of shower variable, and 10% for the free access to main entrance of the site by the rendering truck. These two latter biosecurity measures, manageable directly on the site, should be prioritized and be part of any intervention strategy designed for PRRSV control.     

Ultrasonographic fetal well-being assessment, neonatal and postpartum findings of cloned pregnancies in cattle: A preliminary study on 10 fetuses and calves

Cloned pregnancies in cattle are considered to be at risk due to a variety of fetal or adnexal abnormalities. Data is lacking concerning the possibility of transabdominal ultrasonography in the assessment of these high risk pregnancies. Transabdominal ultrasonography has rarely been reported in the assessment of bovine cloned pregnancies. Ten Holstein heifers carrying 8-month-old cloned fetuses were assessed by transabdominal ultrasonographic examination during the 3rd trimester of pregnancy. Fetal heart rates (FHR), movements, adnexal appearance, and placentome size were recorded. The outcome of the pregnancies was also noted and potential indicators of fetal demise recorded. Survival rate 1 week after birth was 30%. Mean FHR was 113 beats per minute (range: 92 to 128 bpm) during the fetal ultrasonography. No correlation between FHR and fetal activity was found. Fetal hyperactivity and imaging of hyperechoic particles in both allantoic and amniotic fluids were possible signs of fetal distress. Despite the size of the fetus and the deep bovine abdomen, fetal transabdominal ultrasonography can be performed in cattle. This preliminary study points to the necessity of further larger studies for defining normal and abnormal findings in bovine late pregnancy.     

Necrotoxic Escherichia coli (NTEC): two emerging categories of human and animal pathogens.

Necrotoxic Escherichia coli (NTEC) were originally defined as strains of E. coli producing a toxin called cytotoxic necrotising factor (CNF). Two types of CNF have been identified, each of them being genetically linked to several other specific virulence markers, a situation that allows the definition of two distinct homogeneous categories of NTEC called NTEC-1 and NTEC-2. CNF1 and CNF2 are highly homologous holoproteins containing 1,014 amino acids that exert both lethal and necrotic activities in vivo and induce multinucleation and actin stress fibres in cell cultures. The activity of CNFs on mammal cells is due to their ability to constitutively activate by deamidation the Rho proteins, a family of small GTPases that regulate the physiology of the cell cytoskeleton. In NTEC-1, the gene encoding CNF1 belongs to a pathogenicity island which also comprises the genes encoding for alpha-haemolysin and P-fimbriae. In NTEC-2 strains, CNF2 is encoded by a plasmid that also encodes, in 100% of the isolates, a new member of the cytolethal distending toxin family (CDT-III) and in about 50% of the isolates, the F17b-fimbrial adhesin that confers the ability to adhere to calf intestinal villi. The presence of CDT is also suspected in a large majority of NTEC-1 strains. NTEC-1 strains can be found in humans and in all species of domestic mammals, whereas NTEC-2 strains have only been reported in ruminants. The implication of NTEC strains has been clearly established in extra-intestinal infections of humans and animals, for instance in urinary tract infections for NTEC-1 strains. Their role in severe dysenteric syndromes, both in humans and animals, is substantiated by several clinical reports, but there is little published information on this pathogenicity in animal models of infection. The combined production of several powerful toxins (haemolysin, CNF, CDT) by NTEC strains makes them, however, potentially aggressive pathogens which deserve to be searched for on a larger scale. Moreover, NTEC-1 from man and animals appear to be highly related according to available molecular markers, which indicates that domestic animals could constitute reservoirs of NTEC strains which are pathogenic for humans.     

Improved assessment of frozen/thawed mouse spermatozoa using fluorescence microscopy

Genetically modified (GM) animals are unique mutants with an enormous scientific potential. Cryopreservation of pre-implantation embryos or spermatozoa is a common approach for protecting these lines from being lost or to store them in a repository. A mutant line can be taken out of a breeding nucleus only if sufficient numbers of samples with an appropriate level of quality are cryopreserved. The quality of different donors within the same mouse line might be heterogeneous and the cryopreservation procedure might also be error-prone. However, only limited amounts of material are available for analysis. To improve the monitoring of frozen/thawed spermatozoa, commonly used in vitro fertilization (IVF) followed by embryo transfer were replaced with animal-free techniques. Major factors for assessing spermatozoa quality (i.e., density, viability, motility, and morphology) were evaluated by fluorescence microscopy. For this, a live/dead cell staining protocol requiring only small amounts of material was created. Membrane integrity was then examined as major parameter closely correlated with successful IVF. These complex analyses allow us to monitor frozen/thawed spermatozoa from GM mice using a relatively simple staining procedure. This approach leads to a reduction of animal experiments and contributes to the 3R principles (replacement, reduction and refinement of animal experiments).     

Impact of dietary tryptophan and behavioral type on behavior, plasma cortisol, and brain metabolites of young pigs.

The behavioral reactivity in an "open-field" test and plasma cortisol levels were studied in 72 pigs from 12 litters fed for 3 wk one of three diets with different levels of tryptophan: deficient (.14%), adequate (.23%), or excess (.32%). "Open-field" tests were performed three times: 5 d (day W + 5), 23 d (day W + 23) and 45 d (day W + 45) after weaning. The exploration time and the number of grunts provided an adequate measure of the individual emotional reactivity at day W + 5. Significant correlations were obtained between exploration time and the number of grunts at each time (r = -.83 at day W + 5; r = -.46 at day W + 23; r = -.71 at day W + 45). The distinction between animals remained (P less than .05) in terms of exploration time at both 23 and 45 d after weaning. At day W + 23, exploration time was lower in the group fed the adequate diet than in the two other groups. This effect was maintained subsequently after feeding all pigs the same adequate diet (day W + 45). In 36 pigs slaughtered at day W + 23, brain TRP concentration was higher with the excess dietary TRP than with deficient or adequate levels. Conversely, other plasma amino acids (particularly threonine) accumulated only in the brains of pigs fed the deficient diet. Plasma cortisol level assayed at weaning (W) and 2 wk later increased with age and was higher in 16-h fasted (day W + 15) than in 3-h fasted (day W + 17) pigs. Correlations were observed within litters in the fasting state, between the cortisol level and behavioral traits measured at day W + 23 (r = .70 for number of grunts, r = -.60 for exploration time). Dietary TRP did not affect the plasma cortisol level irrespective of the nutritional state after weaning. However, an interaction was noted between plasma cortisol and TRP status (P less than .05). Although dietary TRP induced large variations in brain amino acids and 5-hydroxyindole concentrations, changes in behavioral and cortisol responses were relatively minor.     

Administration of certain sedative drugs is associated with variation in sonographic and radiographic splenic size in healthy cats

Ultrasonography and radiography are standard diagnostic tests for cats with suspected splenic disease, however published information on outside sources of variation are currently lacking. The purpose of this prospective, randomized, crossover group study was to evaluate effects of common sedative drugs on the sonographic and radiographic characteristics of the spleen in healthy cats. Fifteen healthy adult research cats were randomly assigned into one of three groups corresponding to different sequences of administration of five sedative drugs/drug combinations (acepromazine; butorphanol; dexmedetomidine; midazolam and butorphanol (MB); and dexmedetomidine, butorphanol, and ketamine (DBK)), administered at 1‐week intervals. At each visit, three‐view abdominal radiographic and ultrasonographic examinations were performed prior to sedation and repeated 15‐30 min and 2‐3 h post sedation. Two board‐certified radiologists (one ACVR and one ACVR/ECVDI) evaluated the anonymized and randomized images. Acepromazine resulted in significantly increased sonographic and radiographic splenic measurements from baseline, which remained significantly increased 2‐3 h post sedation. The mean magnitude of this change ranged from 0.9 mm (tail height, SD 1.4 mm) to 1.8 mm (body height, SD 1.7 mm) for ultrasound, and was 2.2 mm (ventrodorsal width, SD 2.3 mm) for radiographs. With butorphanol, there was no significant change in splenic size. For dexmedetomidine, MB, and DBK, there was a trend toward increased splenic size from baseline to the first post‐sedation timepoint, which was statistically significant for radiographic measurements, although not for ultrasound. Findings indicated that acepromazine should be avoided prior to imaging while butorphanol may be used when sedation is needed in cats presenting for potential splenic disease.     

A longitudinal study of the risks for introduction of severe footrot into sheep flocks in the south west of Norway

In 2008, ovine footrot was detected in Norway for the first time since 1948. By December 2012 it had spread to 99 flocks, all in the county of Rogaland in the south west of Norway, and 42% of which were located in the municipality of Rennesøy in Rogaland. The aim of this study was to investigate risk factors for contracting severe footrot in flocks of sheep. A flock was considered positive for severe footrot based on positive virulence test or by clinical signs in addition to a positive PCR test.     

Two promoter rearrangements in a drug efflux transporter gene are responsible for the appearance and spread of multidrug resistance phenotype MDR2 in Botrytis cinerea isolates in French and German vineyards

In French and German vineyards, Botrytis cinerea isolates with multiple fungicide resistance phenotypes have been observed with increasing frequencies. Multidrug resistance (MDR) results from mutations that lead to constitutive overexpression of genes encoding drug efflux transporters. In MDR2 and MDR3 strains, overexpression of the major facilitator superfamily transporter gene mfsM2 has been found to result from a rearrangement in the mfsM2 promoter (type A), caused by insertion of a retroelement (RE)-derived sequence. Here, we report the discovery of another, similar RE-induced rearrangement of the mfsM2 promoter (type B) in a subpopulation of French MDR2 isolates. MDR2 isolates harboring either type A or type B mutations in mfsM2 show the same resistance phenotypes and similar levels of mfsM2 overexpression. RE sequences similar to those in mfsM2 were found in low copy numbers in other but not all B. cinerea strains analyzed, including non-MDR2 strains. Population genetic analyses support the hypothesis that the two rearrangement mutations have only occurred once, and are responsible for the appearance and subsequent spread of all known MDR2 and MDR3 strains in French and German wine-growing regions.