Frank Gasthuys

High occurrence of methicillin-resistant Staphylococcus aureus ST398 in equine nasal samples

Methicillin-resistant Staphylococcus aureus (MRSA) infections do occur in equine patients. Little is known, however, about their origin and the general equine MRSA colonization status. In West European horses in particular, neither the colonization rate nor the present strains or their antimicrobial susceptibility patterns are known. In the present study, a sample of 110 (Belgian, French, Dutch and Luxemburg) horses presented at a Belgian equine clinic was screened for nasal MRSA carriage. An indirect culturing protocol using a 0.001% colistin and nalidixic acid containing broth was compared to a direct agar method. Phenotypic identification following growth on a chromogenic MRSA screening agar (ChromID MRSA) was combined with genotypic analysis (PCR, PFGE, SCCmec, spa, and MLST typing). Antimicrobial susceptibility was tested through disk diffusion. Twelve (10.9%) horses carried MRSA, with the enrichment protocol resulting in a significantly higher isolation rate. None of the isolated strains were typeable through SmaI PFGE. They all harboured SCCmec type IVa or V and belonged to spa type t011 or t1451 of the ST398 lineage. All isolates were tetracycline resistant and sulfonamide and enrofloxacin susceptible. Macrolide, lincosamide, trimethoprim and aminoglycoside susceptibility varied and in total five different antimicrobial resistance patterns were distinguished. These results show that ST398 is certainly present in West European horses. Due to its known interspecies transmission and the structure of the equine industry, the presence of this clone in horses poses a substantial health hazard for both animals and humans.

Accuracy of pressure plate kinetic asymmetry indices and their correlation with visual gait assessment scores in lame and nonlame dogs

OBJECTIVE To determine the accuracy of pressure plate kinetic asymmetry indices (ASIs) for diagnosis of unilateral hind limb lameness in dogs and their correlation with visual gait assessment (VGA) scores. ANIMALS 9 healthy dogs and 16 dogs with previously diagnosed unilateral rupture of the cranial cruciate ligament and concurrent unilateral hind limb lameness. PROCEDURES Dogs were walked over a pressure plate to determine paw contact area (PCA), peak vertical pressure (PVP), peak vertical force (PVF), and vertical impulse (VI) of both hind limbs. An ASI was calculated for each gait variable. Simultaneously, gait was assessed visually and scored by use of a numeric rating scale (0 to 10). The ASI of each variable was tested for its usefulness in discrimination between lame and nonlame dogs and for correlation with VGA scores. RESULTS Sensitivity and specificity of ASIs to discriminate between lame and nonlame dogs were excellent for PVF, VI, and PCA; these values were substantially lower for ASI of PVP. Cutoff values to discriminate between lame and nonlame dogs were determined by use of ASIs for PVF, VI, and PCA; however, this could not be done for ASI of PVP. Correlations between ASIs of PVF, VI, and PCA and VGA scores were higher than correlation between the ASIs of PVP and VGA scores. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that ASIs of PVF and VI determined via analysis of pressure plate measurements were reliable indicators of clinical lameness in dogs, but the ASI of PVP was not. The ASI of PCA is an interesting new variable for assessment of limb loading symmetry.

Influence of a constant rate infusion of dexmedetomidine on cardiopulmonary function and recovery quality in isoflurane anaesthetized horses

OBJECTIVE To investigate the influence of a dexmedetomidine constant rate infusion (CRI) in horses anaesthetized with isoflurane. STUDY DESIGN Prospective, randomized, blinded, clinical study. ANIMALS Forty adult healthy horses (weight mean 491 ± SD 102 kg) undergoing elective surgery. METHODS After sedation [dexmedetomidine, 3.5 μg kg(-1) intravenously (IV)] and induction IV (midazolam 0.06 mg kg(-1), ketamine 2.2 mg kg(-1)), anaesthesia was maintained with isoflurane in oxygen/air (FiO(2) 55-60%). Horses were ventilated and dobutamine was administered when hypoventilation [arterial partial pressure of CO(2) > 8.00 kPa (60 mmHg)] and hypotension [arterial pressure 70 mmHg] occurred respectively. During anaesthesia, horses were randomly allocated to receive a CRI of dexmedetomidine (1.75 μg kg(-1) hour(-1) ) (D) or saline (S). Monitoring included end-tidal isoflurane concentration, cardiopulmonary parameters, and need for dobutamine and additional ketamine. All horses received 0.875 μg kg(-1) dexmedetomidine IV for the recovery period. Age and weight of the horses, duration of anaesthesia, additional ketamine and dobutamine, cardiopulmonary data (anova), recovery scores (Wilcoxon Rank Sum Test), duration of recovery (t-test) and attempts to stand (Mann-Whitney test) were compared between groups. Significance was set at p < 0.05. RESULTS Heart rate and arterial partial pressure of oxygen were significantly lower in group D compared to group S. An interaction between treatment and time was present for cardiac index, oxygen delivery index and systemic vascular resistance. End-tidal isoflurane concentration and heart rate significantly increased over time. Packed cell volume, systolic, diastolic and mean arterial pressure, arterial oxygen content, stroke volume index and systemic vascular resistance significantly decreased over time. Recovery scores were significantly better in group D, with fewer attempts to stand and significantly longer times to sternal position and first attempt to stand. CONCLUSIONS AND CLINICAL RELEVANCE A dexmedetomidine CRI produced limited cardiopulmonary effects, but significantly improved recovery quality.

A pressure plate study on fore and hindlimb loading and the association with hoof contact area in sound ponies at the walk and trot

The aim of this study was to evaluate the association between fore- and hind-hoof contact area and limb loading. Data from a previous study on forelimb loading and symmetry were compared with data on hindlimb kinetics, and the fore- and hind-hoof contact area at the walk and trot was evaluated. Five sound ponies, selected for symmetrical feet, were walked and trotted over a pressure plate embedded in a custom-made runway. The hindlimb peak vertical force (PVF) and vertical impulse (VI) were found to be significantly lower than in the forelimb, whereas their high symmetry ratios (>95%) did not show a significant difference from forelimb data. Hindlimb PVF in ponies was found to be slightly higher when compared to data reported for horses even though the ponies moved at a similar or lower relative velocity. The contact area had low intra-individual variability and was significantly smaller in the hind- than in the fore-hooves. A larger contact area was significantly associated with lower peak vertical pressure (PVP) but higher PVF and VI. No significant differences between left and right sides were found for contact area or loading variables. Pressure plate measurements demonstrated a significant association between hoof contact area and limb loading, in addition to intrinsic differences between fore and hindlimb locomotor function. The pressure plate provides the clinician with a tool to quantify simultaneously contralateral differences in hoof contact area and limb loading.

Comparison of pressure plate and force plate gait kinetics in sound Warmbloods at walk and trot

Modern pressure plates (PP) could be an alternative to traditional force plates (FP) for quantitative equine gait analysis, thereby providing the clinician with objective data on the horses gait while unravelling the loading of different regions of the hoof during the stance phase. The aim of this study was to determine whether a stand-alone PP allows reliable measurement of gait kinetics, compared to simultaneously recorded FP variables. Six sound Warmblood horses were walked and trotted over a combined PP and FP system for collection of a set of five valid kinetic measurements for each forelimb. A measurement was considered valid if the horse was moving in a straight line at a constant pace while gait velocity was within a preset range and the hoof fully contacted the plate surface. Significant differences between FP and PP data were seen for peak vertical force (PVF), vertical impulse (VI), time at which the PVF occurs (tPVF) and forelimb symmetry ratios (SymPVF and SymVI) (P < 0.05), but not for stance phase duration (ST). Nevertheless, mean agreement indices (AIs) of ST, tPVF and SymPVF and SymVI were excellent (≥ 0.92), whereas AIs of PVF and VI were moderate (≥ 0.70). The excellent agreement between PP and FP symmetry ratios confirms that observed differences between PP and FP in symmetry ratios are small (2-7%), especially when compared to the expected decrease in symmetry associated with mild lameness (> 20%). The results indicate that a stand-alone pressure plate can be used to measure absolute (ST) and relative (tPVF) temporal variables and loading symmetry ratios and offers equine veterinarians a mobile, cost-efficient and quick gait evaluation method for routine clinical use. However, the system cannot be used interchangeably with a force plate to measure absolute values of limb loading.

A preliminary study on the effects of atropine sulphate on bradycardia and heart blocks during romifidine sedation in the horse.

Romifidine (STH 2130-Cl or Sedivet) is an ά2-agonistic imino-imidazol sedative for intravenous use in horses recently developed by Boehringer Ingelheim, Vetmedica GmbH. An exploratory study was done in nine warm-blood horses, randomly divided into three groups, which received different dosages of romifidine (0.04, 0.08 and 0.12 mg/kg of body weight (BWT) intravenously (i.v.)) with at least one weeks interval between tests.Romifidine induced a marked bradycardia accompanied by second degree atrioventricular (AV) block and some sinus blocks at all tested dosages. A placebo (NaCl 0.9% i.v.) given 5 min before and after romifidine did not affect the cardiac disturbances induced by romifidine.A low dose of atropine sulphate (0.005 mg/kg of BWT i.v.) given 5 min before romidifine counteracted the bradycardia and caused a normal to increased heart rhythm at all romifidine dosages. A higher dose of atropine sulphate (0.01 mg/kg of BWT i.v.) administered 5 min before sedation induced a tachycardia (average 70 beats/min) at all romifidine dosages and completely prevented the bradycardia and the heart blocks. The positive chronotrope effects of atropine sulphate were attenuated by increasing doses of romifidine.The effects of atropine sulphate (low or high doses) given 5 min after romifidine only appeared after 5 min. Both dosages counteracted the bradycardia and suppressed the heart blocks.No atropine-dependent side effects were observed in non-fasted horses. The degree of the romifidine induced sedation was not affected by the use of atropine sulphate given before or after romifidine.

Role of Heat-Stable Enterotoxins in the Induction of Early Immune Responses in Piglets after Infection with Enterotoxigenic Escherichia coli

Enterotoxigenic Escherichia coli (ETEC) strains that produce heat-stable (ST) and/or heat - labile (LT) enterotoxins are cause of post – weaning diarrhea in piglets. However, the relative importance of the different enterotoxins in host immune responses against ETEC infection has been poorly defined. In the present study, several isogenic mutant strains of an O149:F4ac+, LT+ STa+ STb+ ETEC strain were constructed that lack the expression of LT in combination with one or both types of ST enterotoxins (STa and/or STb). The small intestinal segment perfusion (SISP) technique and microarray analysis were used to study host early immune responses induced by these mutant strains 4 h after infection in comparison to the wild type strain and a PBS control. Simultaneously, net fluid absorption of pig small intestinal mucosa was measured 4 h after infection, allowing us to correlate enterotoxin secretion with gene regulation. Microarray analysis showed on the one hand a non-toxin related general antibacterial response comprising genes such as PAP, MMP1 and IL8. On the other hand, results suggest a dominant role for STb in small intestinal secretion early after post-weaning infection, as well as in the induced innate immune response through differential regulation of immune mediators like interleukin 1 and interleukin 17.

Haemodynamic changes during sedation in ponies.

The cardiovascular changes induced by several sedatives were investigated in five ponies with a subcutaneously transposed carotid artery by means of cardiac output determinations (thermodilution technique), systemic and pulmonary artery pressure measurements (direct intravascular method) and arterial blood analysis (blood gases and packed cell volume).The cardiovascular depression (decrease in systemic blood pressure and cardiac output) was long lasting (>90 min) after administration of propionylpromazine (0.08 mg/kg intravenous (i.v.)) together with promethazine (0.08 mg/kg i.v.). The phenothiazine-induced sedation was not optimal.α2-Agonists (xylazine (0.60 mg/kg i.v.) and detomidine (20 μg/kg i.v.)) induced initial but transient cardiovascular effects with an increase in systemic blood pressure and a decrease in cardiac output for about 15 min. Second degree atrioventricular blocks and bradycardia were seen during this period. The cardiovascular depression was more pronounced during detomidine sedation.Atropine (0.01 mg/kg i.v.) induced a tachycardia with a decrease in stroke volume but did not alter the cardiac output or other cardiovascular parameters. It prevented the occurrence of the bradycardia and heart blocks normally induced by xylazine or detomidine. Atropine potentiated the initial hypertension induced by the α2-agonists sedatives (especially detomidine). The decrease in cardiac output induced by xylazine, and to a lesser extent by detomidine, was partially counteracted when atropine was given in advance.The atropine-xylazine combination seemed the best premedication protocol before general anaesthesia as it only resulted in minor and transient cardiovascular changes.

Effects of a constant rate infusion of detomidine on cardiovascular function, isoflurane requirements and recovery quality in horses

OBJECTIVE To examine the influence of a detomidine constant rate infusion (CRI) on cardiovascular function, isoflurane requirements and recovery quality in horses undergoing elective surgery. STUDY DESIGN Prospective, randomized, blinded, clinical trial. ANIMALS Twenty adult healthy horses. METHODS After sedation (detomidine, 10 μg kg(-1) intravenously [IV]) and induction of anaesthesia (midazolam 0.06 mg kg(-1) , ketamine 2.2 mg kg(-1) IV), anaesthesia was maintained with isoflurane in oxygen/air (inspiratory oxygen fraction 55%). When indicated, the lungs were mechanically ventilated. Dobutamine was administered when MAP<70 mmHg. The horses were randomly allocated to one of two groups and throughout anaesthesia, received either a detomidine (5 μg kg(-1)  hour(-1) ) (D) or saline (S) CRI, with the anaesthetist unaware of the treatment. Monitoring included end-tidal isoflurane concentration, arterial pH, PaCO(2) , PaO(2) , dobutamine administration rate, heart rate (HR), arterial pressure, cardiac index (CI), systemic vascular resistance (SVR), stroke index and oxygen delivery index (ḊO(2) I). For recovery from anaesthesia, all horses received 2.5 μg kg(-1) detomidine IV. Recovery quality and duration were recorded in each horse. For statistical analysis, anova, Pearson chi-square and Wilcoxon rank sum tests were used as relevant. RESULTS Heart rate (p=0.0176) and ḊO(2) I (p= 0.0084) were lower and SVR higher (p=0.0126) in group D, compared to group S. Heart rate (p=0.0011) and pH (p=0.0187) increased over time. Significant differences in isoflurane requirements were not detected. Recovery quality and duration were comparable between treatments. CONCLUSIONS AND CLINICAL RELEVANCE A detomidine CRI produced cardiovascular effects typical for α(2) -agonists, without affecting isoflurane requirements, recovery duration or recovery quality.

Use of blood culture medium enrichment for synovial fluid culture in horses: a comparison of different culture methods.

REASONS FOR PERFORMING STUDY Standard methods for culturing equine synovial fluid (SF) are often unrewarding. Evidence-based information on the relative efficiency of different systems used for optimisation of isolation of microorganisms from equine SF is lacking. OBJECTIVES To compare the results of different culture systems performed in parallel on SF samples from horses clinically diagnosed with synovial sepsis. METHODS Synovial fluid specimens were collected between February 2007 and October 2008 from all horses admitted to a referral hospital that were clinically diagnosed with synovial sepsis and from control horses. Synovial fluid samples were cultured in parallel by: 1) direct agar culture (DA); agar culture after: 2) lysis-centrifugation pretreatment (LC); 3) conventional enrichment (CE); 4) combined LC/CE; or 5) blood culture medium enrichment using an automated system (BACTEC 9050). RESULTS Ninety SF samples from 82 horses were included, together with 40 control samples. Seventy-one of 90 samples (79%) were culture-positive by using blood culture medium enrichment (BACTEC), which was significantly higher compared to all other methods. BACTEC enrichment was never negative while any of the other methods was positive. Although agar culture following LC and/or CE resulted in a slightly higher number of positive samples compared to DA, this difference was not significant. All control samples were culture negative by the 5 different techniques. Although the majority of samples containing isolates recovered without enrichment, culture results after BACTEC enrichment were available on the same day as for agar culture with or without LC (19/23 samples), while CE postponed recovery by at least one day in 20/23 samples. CONCLUSION Blood culture medium enrichment is superior to other techniques for isolation of bacteria from SF of horses. The use of an automated system allows enrichment without substantially postponing recovery of microorganisms. POTENTIAL RELEVANCE The efficient and fast isolation of microorganisms from infected SF by the BACTEC system allows for rapid susceptibility testing and a more appropriate antibiotic treatment.