MRSA CC398 in the pig production chain

In 2005, a distinct clone of methicillin resistant Staphylococcus aureus (MRSA CC398) was found in pigs and people in contact with pigs. The structure of the pig production chain in high technology pig husbandry enables pathogens to spread during animal trading, with an increasing prevalence in herds further down the chain. The objective of this study was to quantify the effect of the MRSA status of the supplying herd on the MRSA status of the receiving herd in order to gain more insight into the role of animal trading as a transmission route for MRSA CC398. Nasal samples (60-80 pigs per herd) were collected from 38 herds; in 20 herds, environmental samples were collected as well. Ten MRSA-positive herds (based on the results of nasal swabs of 10 individual pigs per herd) from a prior study were included in the data analysis. Herds were classified as MRSA positive if at least one sample tested positive. The 48 herds were part of 14 complete (40 herds) and 4 incomplete (8 herds) pig production chains. Fifty-six percent of the herds were classified as MRSA positive. MRSA-positive herds were observed at the start (breeding herds), middle (farrowing herds) and the end (finishing herds) of the pig production chain. All of the herds in 8 chains tested MRSA positive;, all of the herds in 5 chains tested MRSA negative and in the remaining 5 chains, MRSA-positive and MRSA-negative herds were detected. Seven spa types were found, which were all previously confirmed to belong to CC398. All of the isolates were susceptible to mupirocin, linezolid, rifampicin, fusidic acid and cotrimoxazole. Resistance against tetracycline, erythromycin and clindamycin was found in 100, 74 and 76% of the isolates, respectively. Seventy-nine percent of herds with a MRSA-positive supplier of pigs were MRSA positive, whereas 23% of herds with a MRSA-negative supplier were MRSA positive (OR=10.8; 95% CI: 1.5-110.1; P=0.011). The presence of entirely MRSA-positive and MRSA-negative chains and the strong association between the MRSA status of herds and their suppliers illustrates a large risk associated with purchasing pigs from MRSA-positive herds; a top-down strategy for future control programs is, therefore, a basic requirement. However, 23% of herds with a MRSA-negative supplier were MRSA positive and furthermore, 46% of the herds at the top of the pig production chain without a supplier tested MRSA positive. This underlined the need for the identification of additional risk factors for MRSA.     

Absence reduction in entomological surveillance data to improve niche-based distribution models for Culicoides imicola

Data-driven models for the prediction of bluetongue vector distributions are valuable tools for the identification of areas at risk for bluetongue outbreaks. Various models have been developed during the last decade, and the majority of them use linear discriminant analysis or logistic regression to infer vector-environment relationships. This study presents a performance assessment of two established models compared to a distribution model based on a promising ensemble learning technique called Random Forests. Additionally, the impact of false absences, i.e. data records of suitable vector habitat that are, for various reasons, incorrectly labelled as absent, on the model outcome was assessed using alternative calibration-validation schemes. Three reduction methods were applied to reduce the number of false absences in the calibration data, without loss of information on the environmental gradient of suitable vector habitat: random reduction and stratified reduction based on the distance between absence and presence records in geographical (Euclidean distance) or environmental space (Mahalanobis distance). The results indicated that the predicted vector distribution by the Random Forest model was significantly more accurate than the vector distributions predicted by the two established models (McNemar test, p<0.01) when the calibration data were not reduced with respect to false absences. The performance of the established models, however, increased considerably by application of stratified false absence reductions. Model validation revealed no significant difference between the performance of the three distinct Culicoides imicola distribution models for the majority of alternative stratified reduction schemes. The main conclusion of this study is that the application of Random Forests, or linear discriminant analysis and logistic regression on the condition that calibration data were first reduced on geographical or environmental information, potentially lead toward better vector distribution models.     

Morphological changes in the small intestinal smooth muscle layers of horses suffering from small intestinal strangulation. Is there a basis for predisposition for reduced contractility?

Reasons for performing study: Intestinal strangulation often leads to enterectomy after which ileus can develop. This has prompted research to look into possible pathophysiological processes triggering equine ileus. However, morphological changes of the small intestinal smooth muscle in relation to equine colic have not yet been studied. Objectives: The presence of some smooth muscle proteins was morphologically assessed and quantified in control and colic horses. In addition, the up‐ or down‐regulation of heat shock proteins (HSP20 and HSP27) influencing the contractility of smooth muscles was studied. Methods: Cranial resection margins of 18 strangulated small intestinal samples were collected. Small intestinal control samples were collected from 11 horses subjected to euthanasia for other than gastrointestinal‐related reasons. Formaldehyde‐fixed tissue was paraffin‐embedded and processed for conventional staining and immunohistochemistry. Snap‐frozen full‐thickness biopsies were collected for western blot analyses. Results: Evaluating the muscle layer microscopically, colic samples showed significantly more signs of degradation than controls (P = 0.026) of which vacuolar degeneration was most prominent (P = 0.009). In colic samples, myosin protein levels were decreased (P = 0.022) whereas desmin (P = 0.049) and HSP20 protein levels (P = 0.005) were elevated. Conclusions: In colic samples, microscopic lesions at the level of the muscle layer indicate a stress response. In addition, modified amounts of structural proteins such as myosin and desmin together with increased HSP20 levels could perhaps provide a basis for explaining the malfunctioning of the intestinal muscle layer. Potential relevance: Post operative ileus, following small intestinal strangulation and resection, could be related in part to a dysfunctional muscle layer. In addition to microscopic signs of degeneration, myosin and HSP20 were affected. Pharmacological interventions might alter HSP20 expressions and thus serve a protective effect.     

Application of ROC curve analysis to FAMACHA© evaluation of haemonchosis on two sheep farms in South Africa

Test sensitivity and specificity for the FAMACHA(?) clinical test for anaemia due to haemonchosis have previously been shown to be adequate in differentiating between heavily/less heavily infected sheep, but these properties give no objective guidance for setting the optimum threshold at which anthelmintic treatment should occur. The aim of this work was to use Receiver Operating Characteristic (ROC) curves to evaluate the diagnostic accuracy of FAMACHA(?) testing by estimating the area under the ROC curve, and to use two-graph ROC curves to decrease subjectivity in selecting treatment thresholds on two farms with contrasting management. Test diagnostic accuracy, and thus discriminating power as determined by the area under the ROC curves, ranged from "moderate to good" on the first farm, and from "moderate to high" on the second farm for haematocrit (the Gold Standard for the test) cut-offs of ≤ 22% and ≤ 19% on both farms respectively. Accuracy of classification between haematocrit cut-offs was not significantly different within farms, but did differ significantly between farms, with test accuracy being highest on the second farm at both haematocrit cut-offs (p<0.05). The results also showed the suitability of the two-graph ROC curve approach for discriminating not only between different levels of accuracy of evaluators, but also to give an indication of the so-called ROC cut point (i.e. the desired threshold level) at which animals should be treated for a given level of risk of loss. The approach appears to have the potential not only to validate the diagnostic accuracy of the test across the complete testing range (i.e. all FAMACHA(?) categories from 1 to 5), but also to compensate for such inaccuracy by allowing objective adjustment of the threshold treatment level according to the output of the two-graph ROC method.     

One year's study of dynamic and evolution of types I and II PRRSV in a swine farm

This study was to investigate dynamic and evolution of PRRSV in a seed-stock farm by monitoring PRRSV status from 11 June 2009 to 4 August 2010. For laboratory test, around 18-24 umbilical cords from farrowed sows and 5-95 sera from nursery and grow/finish pigs were submitted around every 2 weeks interval during the study. The submitted samples were tested for PRRSV using IDEXX PRRS 2XR ELISA kit, RT-nested PCR. The PRRSV-positive samples were further sequences based on ORF5 and analyzed using MEGA 3.1 program and Beast 1.5.4 package. The surveyed farm was first infected with type II PRRSV but it was infected newly with type I PRRSV of unknown origin, showing rapid substitution to type I PRRSV as a dominant strain in 2 weeks. The type I PRRSV was first detected from umbilical cord of a farrowed sow in 12 January 2010, and secondly from nursery pigs in 26 January 2010. Although sudden increase of mean S/P ratio was found in grow/finish pigs around 2 months earlier than first type I PRRSV detection, no type I PRRSV viremia was found. Thirty three ORF5 full sequences from 14 type II to 19 type I PRRSVs were obtained chronologically in this farm and the genetic characteristics and evolution rates of those sequences were analyzed. The substitution rates (/site/day) of two types were 4.03×10(-5) (type I), 3.09×10(-5) (type II), respectively, which was more frequent than previous reports. The calculated divergence time of type I PRRSV was consistent with the time when the sudden elevation of serum IgG in grow/finish barn was first observed. This study provided fundamental data for type I PRRSV dynamic in a previously type II PRRSV-infected farm and suggested grow/finisher barn could be a primary site for PRRSV introduction.     

Effect of the US3 protein of bovine herpesvirus 5 on the actin cytoskeleton and apoptosis

The US3 protein is a unique protein kinase only present in the Alphaherpesvirinae subfamily of the herpesviruses. Studies performed with several alphaherpesviruses demonstrated that the US3 protein is involved in cytoskeleton modifications during viral infection and displays anti-apoptotic activity. However, the US3 protein of BoHV-5 has not been studied up to now. As reported for other alphaherpesviruses, our results showed that BoHV-5 US3 confers resistance against apoptosis and induces cytoskeletal reorganization leading to cell rounding, actin stress fiber breakdown and cell projections that interconnect cells. The expression of a kinase-dead version of BoHV-5 US3 showed that the anti-apoptotic activity and the induction of cell projections are kinase-dependent whereas kinase activity is not absolutely required for actin stress fiber breakdown. Besides, the kinase-dead version of US3, but not the wild type protein, was found excluded from the nucleus. These results constitute the first report on the BoHV-5 US3 functions, and highlight that there are functional differences and similarities among US3 proteins of different alphaherpesviruses.     

Angiotensin I-converting enzyme inhibitory activity of gelatin hydrolysates and identification of bioactive peptides

In this project we report on the angiotensin I-converting enzyme (ACE)-inhibitory activity of a bovine gelatin hydrolysate (Bh2) that was submitted to further hydrolysis by different enzymes. The thermolysin hydrolysate (Bh2t) showed the highest in vitro ACE inhibitory activity, and interestingly a marked in vivo blood pressure-lowering effect was demonstrated in spontaneously hypertensive rats (SHR). In contrast, Bh2 showed no effect in SHR, confirming the need for the extra thermolysin hydrolysis. Hence, an angiotensin I-evoked contractile response in isolated rat aortic rings was inhibited by Bh2t, but not by Bh2, suggesting ACE inhibition as the underlying antihypertensive mechanism for Bh2t. Using mass spectrometry, seven small peptides, AG, AGP, VGP, PY, QY, DY and IY or LY or HO-PY were identified in Bh2t. As these peptides showed ACE inhibitory activity and were more prominent in Bh2t than in Bh2, the current data provide evidence that these contribute to the antihypertensive effect of Bh2t.     

Sedimentary and diagenetic features in saline lake deposits of the Monegros region, northern Spain

The Monegros region in northern Spain is marked by the occurrence of a large number of ephemeral to dry lake basins, occupying small karstic depressions. The lacustrine sediment fill of these basins contain various carbonate and silicate minerals whose origin and palaeoenvironmental significance is poorly understood. For the present study, 14 lake basins were sampled in order to establish vertical, lateral and regional variations in mineralogical and textural characteristics, aimed at determining the mode of formation of the various mineral phases present. In nearly all basins, the same sequence of three lithological units is recognized, including a basal clayey unit, a middle magnesite-bearing and gypsum-rich unit, and a calcite- and dolomite-dominated surface unit. Distribution patterns of carbonate minerals indicate that magnesite is a synsedimentary precipitate, dolomite formed as a diagenetic authigenic phase, and calcite is partly authigenic and partly allogenic. All clay minerals, including sepiolite and smectite, appear to be allogenic. Regional variations are marked by similarities between groups of neighbouring basins, but no overall trend related to regional drainage patterns is recognized. The middle lithological unit records a lake stage with predominantly chemical sedimentation (Unit II), overlying a less well documented interval corresponding to a perennial lake stage with lower salinity (Unit III), whereas the surface unit formed during a period with predominantly clastic sedimentation. Based on a comparison with other regional records, the middle unit is attributed to an Early Holocene humid stage, separated from the overlying Late Holocene deposits by a hiatus that corresponds to a Mid Holocene arid stage.     

Oxygen exchange with water alters the oxygen isotopic signature of nitrate in soil ecosystems

Combined oxygen (O) and nitrogen (N) stable isotope analyses are commonly used in the source determination of nitrate . The source and fate of are studied based on distinct O and N isotopic signatures (δ¹⁸O and δ¹⁵N) of various sources and isotopic effects during transformation processes, which differ between sources like fertilizer, atmospheric deposition, and microbial production (nitrification). Isotopic fractionation during production and consumption of further affects the δ¹⁸O and δ¹⁵N signal. Regarding the δ¹⁸O in particular, biochemical O exchange between O from and H₂O is implicitly assumed not to affect the δ¹⁸O signature of . This study aimed to test this assumption in soil-based systems. In a short (24 h) incubation experiment, soils were treated with artificially ¹⁸O and ¹⁵N enriched . Production of from nitrification during the incubation would affect both the ¹⁸O and the ¹⁵N enrichment. Oxygen exchange could therefore be studied by examining the change in ¹⁸O relative to the ¹⁵N. In two out of the three soils, we found that the imposed ¹⁸O enrichment of the declined relatively more than the imposed enrichment. This implies that O exchange indeed affected the O isotopic signature of , which has important implications for source determination studies. We suggest that O exchange between and H₂O should be taken into consideration when interpreting the O isotopic signature to study the origin and fate of in ecosystems.