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Analysis of salivary cortisol is increasingly used to assess stress responses in horses. Because spontaneous or experimentally induced increases in cortisol concentrations are often relatively small for stress studies, proper controls are needed. This requires an understanding of the factors affecting salivary cortisol over longer times. In this study, we have analyzed salivary cortisol concentration for 6 mo in horses (n = 94) differing in age, sex, reproductive state, and housing. Salivary cortisol followed a diurnal rhythm with the highest concentrations in the morning and a decrease throughout the day ( P < 0.001). This rhythm was disrupted in individual groups on individual days; however, alterations remained within the range of diurnal changes. Comparison between months showed highest cortisol concentrations in December ( P < 0.001). Cortisol concentrations increased in breeding stallions during the breeding season ( P < 0.001). No differences in salivary cortisol concentrations between nonpregnant mares with and without a corpus luteum existed. In stallions, mean daily salivary cortisol and plasma testosterone concentrations were weakly correlated ( r = 0.251, P < 0.01). No differences in salivary cortisol between female and male young horses and no consistent differences between horses of different age existed. Group housing and individual stabling did not affect salivary cortisol. In conclusion, salivary cortisol concentrations in horses follow a diurnal rhythm and are increased in active breeding sires. Time of the day and reproductive state of the horses are thus important for experiments that include analysis of cortisol in saliva. [ABSTRACT FROM AUTHOR]

Changing temperature and precipitation can strongly influence plant reproduction. However, also biotic interactions might indirectly affect the reproduction and recruitment success of plants in the context of climate change. Information about the interactive effects of changes in abiotic and biotic factors is essential, but still largely lacking, to better understand the potential effects of a changing climate on plant populations. Here we analyze the regeneration from seeds of Acer platanoides and Acer pseudoplatanus , two currently secondary forest tree species from seven regions along a 2200 km-wide latitudinal gradient in Europe. We assessed the germination, seedling survival and growth during two years in a common garden experiment where temperature, precipitation and competition with the understory vegetation were manipulated. A. platanoides was more sensitive to changes in biotic conditions while A. pseudoplatanus was affected by both abiotic and biotic changes. In general, competition reduced ( in A. platanoides ) and warming enhanced ( in A. pseudoplatanus ) germination and survival, respectively. Reduced competition strongly increased the growth of A. platanoides seedlings. Seedling responses were independent of the conditions experienced by the mother tree during seed production and maturation. Our results indicate that, due to the negative effects of competition on the regeneration of A. platanoides , it is likely that under stronger competition (projected under future climatic conditions) this species will be negatively affected in terms of germination, survival and seedling biomass. Climate-change experiments including both abiotic and biotic factors constitute a key step forward to better understand the response of tree species’ regeneration to climate change. [ABSTRACT FROM AUTHOR]

Identification of horses by microchip transponder is mandatory within the European Union with only a few exceptions. In this study, the readability of such microchips in 428 horses with three different scanners (A, B and C) and the histological changes at the implantation site in 16 animals were assessed. Identification of microchips differed between scanners (P < 0.001), and with 'side of neck' (P < 0.001). Scanners A, B and C identified 93.5%, 89.7% and 100% of microchips, respectively, on the 'chip-bearing' side of the neck. From the contralateral side, scanners A, B and C identified 21.5%, 26.9% and 89.5% of transponders, respectively. Microchip readability was affected by age (P < 0.001), but not by breed of horse. At necropsy, transponders were found in the subcutaneous fat (n = 3), inter- or peri-muscular connective tissue (n = 8), or musculature (n = 5), where they were surrounded by a fibrous capsule ranging in thickness from 12.7 to 289.5 µm in 15 animals. In two animals, immature granulation tissue with attendant granulomatous inflammation, and a granulomatous myositis, surrounding the microchip were identified, respectively. Severe (n - 1), moderate (n = 1), and mild (n - 3) lymphohistiocytic inflammation was noted within the fibrous capsule. Microchip transponders were found to be a highly reliable and biocom-patible method of horse identification. [ABSTRACT FROM AUTHOR]

Clin Microbiol Infect 2012; 18: 147-152 Abstract In order to perform a cost-effective search and destroy policy for methicillin-resistant Staphylococcus aureus (MRSA), a quick and reliable typing method is essential. In an area with a high level of animal-related MRSA ST398, pulsed field gel electrophoresis (PFGE) typing and spa-typing are not sufficient to discriminate between co-incidental findings and true transmission of MRSA. This study is the first to retrospectively show the performance of Raman spectroscopy in 16 well-documented outbreaks. We analysed 525 isolates, 286 MRSA ST398 and 239 from other PFGE clusters with Raman spectroscopy. When epidemiologically linked isolates from the outbreaks were analysed with PFGE as the reference standard, Raman spectroscopy correctly identified 97% of cases that were indistinguishable from the index case. With Raman cluster analysis, the most dominant distinction was between MRSA ST398 and other MRSA of human clonal lineages. Within MRSA ST398, 22 different Raman clusters were identified. Raman typing correctly identified an ST398 ( spa type t567) outbreak in a hospital setting. No direct correlation was observed between Raman clusters and spa types. We conclude that Raman spectroscopy is a quick and reliable method of MRSA typing, which can be used in outbreak settings and it is comparable to PFGE, with the added advantage that PFGE non-typeable isolates can also be readily typed using the same sample preparation protocol. [ABSTRACT FROM AUTHOR]

Branding is the traditional and well-established method used to mark horses, but recently microchip transponders for implantation have become available. In this study, behaviour, physiological stress variables and skin temperature in foals were determined in response to hot-iron branding (n = 7) and microchip implantation (n = 7). Salivary cortisol concentrations increased in response to branding (1.8 ± 0.2 ng/mL) and microchip implantation (1.4 ± 0.1 ng/mL), but cortisol release over time did not differ. In response to both manipulations there was a transient increase in heart rate (P < 0.001) and heart rate variability (P < 6.01). Branding and microchip implantation induced a comparable aversive behaviour (branding, score 3.86 ± 0.85; microchip, score 4.00 ± 0.82). Both techniques thus caused similar physiological and behavioural changes indicative of stress. Acutely, implantation of a microchip was as stressful as branding in foals. Branding caused a necrotising skin burn lasting at least 7 days. Moreover branding, but not microchip implantation (P < 0.001), was accompanied by a generalized increase in skin temperature which was comparable to low degree post-burn hypermetabolism in humans. [ABSTRACT FROM AUTHOR]

Abstract: A series of monoacylated glycolipids with even-numbered acyl chain lengths ranging from saturated C11 to C15 and an unsaturated C17:1 fatty acid connected by an amide in linkage to the disaccharide head groups maltose, melibiose and lactose were synthesized. The structural polymorphism of the glycolipids was investigated using Fourier-transform infrared spectroscopy and differential scanning calorimetry for the detection of the gel to liquid-crystalline acyl chain melting behaviour and small-angle X-ray scattering for the elucidation of the physical structure of the lipid aggregates. Also, the phase morphology was studied by polarizing microscopy in contact preparations. The data clearly show the existence of uni- and multilamellar structures. Although only one acyl chain is present, there is no evidence for the existence of micelles – of spherical or of cylindrical (HI) type – or of interdigitated phases. The preference for lamellar phases seems to be correlated with the intrinsic high conformational order of the amide linkage of these compounds which inhibits the formation of highly curved structures. [Copyright &y& Elsevier]

Pig farmers and veterinarians in contact with livestock in The Netherlands have a higher risk of methicillin-resistant Staphylococcus aureus (MRSA) carriage than the general population. The objective of this study was to investigate whether this is also true for other professionals in contact with pigs in an international setting. A convenience sample of 272 participants at an international conference on pig health in Denmark was screened for MRSA carriage using combined nose/throat swabs and were asked to complete a questionnaire concerning animal contacts, exposure to known MRSA risk-factors, and the protective measures taken when entering pig farms. In total, 34 (12.5%) participants from nine countries carried MRSA. Thirty-one of these isolates were non-typeable by pulsed-field gel electrophoresis following SmaI digestion of chromosomal DNA. All of the non-typeable isolates belonged to spa types (t011, t034, t108, t571, t567 and t899) that correspond to multilocus sequence type 398. All of the above-mentioned spa types, with the exception of t899, have been isolated previously from either Dutch pigs, pig farmers and/or veterinarians. Protective measures, e.g., masks, gowns and gloves, did not protect against MRSA acquisition. Transmission of MRSA from pigs to staff tending to these animals appears to be an international problem, creating a new reservoir for community-acquired MRSA (CA-MRSA) in humans in Europe, and possibly worldwide. The rise of a new zoonotic source of MRSA could have a severe impact on the epidemiology of CA-MRSA, and may have consequences for the control of MRSA, especially in those countries that maintain a low prevalence by means of search-and-destroy policies. [ABSTRACT FROM AUTHOR]

Abstract: In vivo and in situ methods were used to determine whether chemical methods designed to protect feed crude protein (CP) against ruminal degradation were also effective in protecting crude fat (CF). Four diets were formulated: two rapeseed diets, consisting of (g/kg dry matter (DM)) 400 grass hay, 250 ground barley grain and 350 untreated (RM) or xylose-treated (RMT) expeller rapeseed meal and two soybean diets, consisting of (g/kg DM) 400 grass hay, 300 ground barley grain and 300 untreated (SB) or formaldehyde-treated (SBT) ground soybeans. Four German Red Pied steers, fitted with a ruminal cannula, were used in a 4×4 Latin square arrangement of treatments and fed once daily at 07:00h. Each treatment period lasted 11 days. Ruminal fluid samples were collected and pH values were measured before feeding (0h) and 2, 3 and 5h after beginning of feeding on day 9. On day 11, the rumens of the steers were completely emptied manually 24h after the last feeding. Additionally, feed samples were incubated in situ in the rumen of the steers for 0, 2, 4, 8, 16, 24, 48, 72 and 336h. In vivo, steers fed the soybean diets had less CF and more DM, organic matter, neutral detergent fibre and acid detergent fibre in the rumen compared to the steers fed the rapesed diets at 24h after feeding. In situ, CF of all feedstuffs extensively disappeared from the bags and no treatment effect occurred. Concentrations of ammonia–N in ruminal fluid were lower for RMT and SBT than for RM and SB, indicating that CP of the treated feeds was protected against ruminal degradation, findings that are consistent with the in situ data. Treating ground soybeans with formaldehyde and expeller rapeseed meal with xylose, did not protect against CF degradation in the rumen, although CP protection was effective. Both in vivo and in situ methods were suitable to assess CP degradation in the rumen and both methods also indicated that chemical treatment had no effect on ruminal CF degradation, but the estimated extent of CF disappearance differed between the in vivo and in situ estimates. [Copyright &y& Elsevier]

A solid phase sintering diffusion (SPSD) method was used at the production of the soft magnetic alloys Fe100-x-ySixPy (0

Screening of pig farmers and pigs in The Netherlands has revealed that >20% of pig farmers and 39% of slaughterhouse pigs are positive for an unusual strain of methicillin-resistant Staphylococcus aureus (MRSA) belonging to sequence type (ST) 398. It is now clear that the emergence of ST398 is not just a Dutch problem, with human infections being described in several European countries, Canada and Singapore. Furthermore, some human isolates have now acquired the genes encoding Panton–Valentine leukocidin. Livestock may become an important source of community-acquired MRSA. A concerted effort on the part of clinicians, infection control practitioners and veterinarians will be required to prevent further spread of this novel strain of MRSA. [ABSTRACT FROM AUTHOR]

Identification of horses has traditionally been facilitated by hot iron branding, but the extent by which branding symbols and numbers can be identified has not been investigated. The local pathological changes induced by branding are also unknown. This study analysed the readability of branding symbols and histomorphological alterations at the branding sites. A total of 248 horses in an equestrian championship were available for identification of symbols and numbers. A further 28 horses, euthanased for other reasons, provided histological examination of the branding site. All except one horse had evidence of histological changes at the brand site, including epidermal hyperplasia, increase of dermal collagenous fibrous tissue and loss of adnexal structures. In two foals, an ulcerative to necrotizing dermatitis was observed and interpreted as a complication of recent branding lesions. Despite the fact that hot iron branding caused lesions compatible with third degree thermal injury, it did not allow unambiguous identification of a large proportion of older horses. While the breed-specific symbol was consistently identified by three independent investigators in 84% of the horses, the double-digit branding number was read correctly by all three investigators in less than 40%. In conclusion, hot iron branding in horses causes lesions compatible with third degree thermal injury but does not always allow identification of horses. [ABSTRACT FROM AUTHOR]

Objectives. Evaluation of a new surface treatment method to obtain a good bond strength between a luting composite and several ceramics.Methods. Specimen preparation and test procedure were done according to ISO 10477 Amendment 1. The surfaces of Empress II, InCeram-Alumina, InCeram-Zirconia and Frialit (ZrO2) were ground under water-cooling with 400 grit grinding paper, afterwards polished with 800 grit and air-dried. Each ceramic material investigated was divided into three groups of 10 specimens each. Group 1 was flame-treated with the PyrosilPen for 2.5 s, group 2 for 5 s and group 3 for 10 s/cm2. After the flame treatment, a methacryl silane was applied followed by a luting composite. Prior to measuring shear bond strength, the specimens were thermocycled 5000 times in a water-bath between +5 and +55 °C. Furthermore, SEM- and Fourier-transform infrared spectrophotometer (FT-IR)-investigations were done. As a control, Empress II etched and silaned was used.Results. Shear bond strength measurements indicated that the optimal treatment time was 5 s/cm2. Regarding this time the following bond strength values between the luting composite and the various ceramics were obtained: Empress II 23 (5) MPa, InCeram-Alumina 23 (5) MPa, InCeram-Zirconia 13 (8) MPa, and Frialit 16 (6) MPa. The control achieved 27 (6) MPa. On all surfaces of the flamed specimens Si could be detected by FT-IR.Significance. The PyrosilPen-Technology is an easy and effective method for surface-treating silicate, aluminum oxide and zirconium oxide ceramics to obtain good bonding to luting composites. [Copyright &y& Elsevier]

Climate projections indicate that temperatures will increase by up to 4.5 °C in Europe by the end of this century, and that more extreme rainfall events and longer intervening dry periods will take place. Climate change will likely affect all phases of the life cycle of plants, but plant reproduction has been suggested to be especially sensitive. Here, using a combination of approaches (soil heaters and different provenances along a latitudinal gradient), we analyzed the regeneration from seeds of Acer platanoides and A. pseudoplatanus , two tree species considered, from a management point of view, of secondary relevance. We studied germination, seedling survival and growth in a full-factorial experiment including warming and changes in watering frequency. Both species responded to warming, watering frequency and seed provenance, with stronger (negative) effects of warming and provenance than of watering frequency. In general, the central provenances performed better than the northernmost and southernmost provenances. We also detected interactive effects between warming, watering frequency and/or seed provenance. Based on these results, both species are expected to show dissimilar responses to the changes in the studied climatic factors, but also the impacts of climate change on the different phases of plant regeneration may differ in direction and magnitude. In general increases in the precipitation, frequency will stimulate germination while warming will reduce survival and growth. Moreover, the frequent divergent responses of seedlings along the latitudinal gradient suggest that climate change will likely have heterogeneous impacts across Europe, with stronger impacts in the northern and southern parts of the species’ distribution ranges. [ABSTRACT FROM AUTHOR]

Abstract: The mechanisms leading to parturition in the horse in many aspects differ from those in other species. Pregnancy is maintained not by progesterone but by 5α-pregnanes and the progestin precursor pregnenolone originates from the fetus. As parturition approaches, the fetal adrenal switches from pregnenolone to cortisol synthesis but it is not known whether cortisol crosses the placenta. We hypothesized that in parallel to fetal cortisol release, cortisol in the maternal circulation increases before foaling and this increase can be determined in both saliva and plasma. In addition, maternal, fetal and neonatal heart rate and heart rate variability were measured. In 25 pregnant mares, saliva for cortisol analysis was collected 4 times daily from 15 days before to 5 days after foaling. In 13 mares, in addition, fetomaternal electrocardiogram (ECG) recordings were made and blood samples for progestin and cortisol analysis were collected once daily. Heart rate (HR) was recorded until 5 days after foaling. The heart rate variability (HRV) variables standard deviation of the beat-to-beat (RR) interval (SDRR) and root mean square of successive RR differences (RMSSD) were calculated. From Days 15 to 4 before parturition, progestin concentration increased (peak 267 ± 42 ng/mL) and decreased thereafter (P < 0.05, day of foaling 113 ± 18 ng/mL). A prepartum increase in maternal cortisol concentrations was evident in blood (P < 0.05) and saliva (P < 0.05) and paralleled the decrease in progestin concentrations. In mares, HR remained constant during the last days of pregnancy but decreased within one day after parturition (P < 0.05) while maternal HRV did not change. In the fetus and neonate, HR increased from before to after birth (P < 0.05) indicating increasing demands on the cardiovascular system with adaptation to extrauterine life. [Copyright &y& Elsevier]

Clin Microbiol Infect 2011; 17: 1704-1710 Abstract Multiple body site screening and pre-emptive isolation of patients at risk for methicillin-resistant Staphylococcus aureus (MRSA) carriage are considered essential for control of nosocomial spread. The relative importance of extranasal screening when using rapid diagnostic testing (RDT) is unknown. Using data from a multicentre study evaluating BD GeneOhm™ MRSA PCR (IDI), Xpert MRSA (GeneXpert) and chromogenic agar, added to conventional cultures, we determined cost-effectiveness assuming isolation measures would have been based on RDT results of different hypothetical screening regimes. Costs per isolation day avoided were calculated for regimes with single or less extensive multiple site RDT, regimes without conventional back-up cultures and when PCR would have been performed with pooling of swabs. Among 1764 patients at risk, MRSA prevalence was 3.3% ( n = 59). In all scenarios the negative predictive value is above 98.4%. With back-up cultures of all sites as a reference, the costs per isolation day avoided were €15.19, €30.83 and €45.37 with 'nares only' screening using chromogenic agar, IDI and GeneXpert, respectively, as compared with €19.95, €95.77 and €125.43 per isolation day avoided when all body sites had been screened. Without back-up cultures costs per isolation day avoided using chromogenic agar would range from €9.24 to €76.18 when costs per false-negative RDT range from €5000 up to €50 000; costs for molecular screening methods would be higher in all scenarios evaluated. In conclusion, in a low endemic setting chromogenic agar screening added to multiple site conventional cultures is the most cost-effective MRSA screening strategy. [ABSTRACT FROM AUTHOR]

Pre-emptive isolation of suspected methicillin-resistant Staphylococcus aureus (MRSA) carriers is considered essential for controlling the spread of MRSA, but noncolonized patients will be isolated unnecessarily as a result of a delay in diagnosis of 3-5 days with conventional cultures. We determined costs per isolation day avoided, and incremental costs of rapid MRSA screening tests when added to conventional screening, but with decisions on isolation measures based on PCR results. A prospective multicentre study evaluating BD GeneOhm MRSA PCR ('IDI') (BD Diagnostics, San Diego, CA, USA), Xpert MRSA ('GeneXpert') (Cepheid, Sunnyvale, CA, USA) and chromogenic agar (MRSA-ID) (bioMérieux, Marcy-l'Etoile, France) was performed in 14 Dutch hospitals. Among 1764 patients at risk, MRSA prevalence was 3.3% (n = 59). Duration of isolation was 19.7 and 16.1 h with IDI and GeneXpert, respectively, and would have been 30.0 and 76.2 h when based on chromogenic agar and conventional cultures, respectively. Negative predictive values (at a patient level) were 99.5%, 99.1% and 99.5% for IDI, GeneXpert and chromogenic agar, respectively. Numbers of isolation days were reduced by 60% and 47% with PCR-based and chromogenic agar-based screening, respectively. The cost per test was ϵ56.22 for IDI, ϵ69.62 for GeneXpert and ϵ2.08 for chromogenic agar, and additional costs per extra isolation day were ϵ26.34. Costs per isolation day avoided were ϵ95.77 (IDI) and ϵ125.43 (GeneXpert). PCR-based decision-making added ϵ153.64 (IDI) and ϵ193.84 (GeneXpert) per patient to overall costs and chromogenic testing would have saved ϵ30.79 per patient. Rapid diagnostic testing safely reduces the number of unnecessary isolation days, but only chromogenic screening, and not PCR-based screening, can be considered as cost saving. [ABSTRACT FROM AUTHOR]

• Blue LED light treatment during winter affected pregnancy outcome in horses. • Gestation was shorter in blue LED light-treated than in control pregnancies. • Height at birth was reduced in foals born to blue LED light-treated mares. • Hair length was reduced in foals born to blue LED light-treated mares. • Blue LED did not affect foal birth weight. Blue light directed at 1 eye advances the equine ovulatory season but may also advance foaling. In this study, effects of blue LED light on pregnancy outcome were assessed. A total of 20 mares with singleton pregnancies were studied over 2 consecutive years in a cross-over design. In 1 year, mares received an extended photoperiod using 50 lux of blue LED light (468 nm) directed at a single eye from 08:00 until 23:00 daily via head-worn light masks starting mid-December and in the other year remained untreated as controls. Gestation was shorter in blue LED light-treated than in control pregnancies (median 333.0 vs 338.5 days, P = 0.036). Foals born to blue LED light-treated mares had lower wither heights (median 103.0 vs 104.5 cm, P = 0.023), similar weights (median 55.8 vs 54.8 kg, P = 0.732) and took less time to stand after birth than control foals (median 35.0 vs 53.5 min, P = 0.036). Foals born to blue LED light-treated mares had reduced hair length compared to controls (median 12.0 vs 20.0 mm, P = 0.009) and hair regrowth in treated mares was reduced (P = 0.036). In conclusion, blue LED light directed at 1 eye advanced foaling and influenced height and hair coat but not weight in foals. [ABSTRACT FROM AUTHOR]

Heart rate (HR), HR variability (HRV) and salivary cortisol concentrations were determined in foals (n = 13) during the perinatal phase and until 5 months of age. In the fetus, HR decreased from 77 ± 3 beats/min at 120 min before birth to 60 ± 1 beats/min at 5 min before birth (P < 0.01). Within 30 min of birth, HR increased to 160 + 9 beats/min (P < 0.01). Salivary cortisol concentrations immediately after birth were 11.9 ±3.6 ng/mL and within 2 h increased to a maximum of 52.5±12.3 ng/mL (P<0.01). In conclusion, increases in HR and salivary cortisol concentrations in foals are not induced during parturition, but occur immediately after birth. [ABSTRACT FROM AUTHOR]