Putative regulation mechanism for the MSTN gene by a CpG island generated by the SINE marker Ins227bp.

BACKGROUND: A single nucleotide polymorphism (SNP) in the first intron of the myostatin gene (MSTN) is associated with aptness of elite Thoroughbreds to race over sprint, middle or long distances. This intronic marker (g.66493737 T ≻ C), a short interspersed nuclear element (SINE) of 227 bp (Ins227bp) insertion polymorphism in the MSTN promoter, and the adjacent SNP BIEC2-417495 have not been studied for their association with racing aptness of the average Thoroughbreds raced in countries with lower status of the racing industry. This study investigated these markers regarding their prevalence and association with performance in common race horses. Markers were genotyped by amplification refractory mutation system-quantitative PCR (ARMS-qPCR) or amplicon melting. Furthermore, we asked whether the Ins227bp marker might theoretically regulate the expression of myostatin by generating a novel target for DNA methylation or by changing binding sites for transcription factors. Putative sites for DNA methylation or binding of transcription factors were predicted by MethPrimer and by the softwares JASPAR, MatInspector and UniPROBE, respectively. RESULTS: Pairwise linkage disequilibrium between g.66493737 T ≻ C and Ins227bp was high (r (2) = 0.93). A lower linkage was determined for g.66493737 T ≻ C and BIEC2-417495 (r (2) = 0.69) as well as for BIEC2-417495 and Ins227bp (r (2) = 0.76). The estimated frequencies for the presence of Ins227bp (I) indel and the C alleles at g.66493737 T ≻ C and BIEC2-417495 were 0.46, 0.47 and 0.43, respectively. Heterozygotes represented the most abundant genotype at each locus. The best racing distance (BRD) was significantly different between the homozygotes of each SNP (p = 0.01 to 0.03). C allele homozygotes at BIEC2-417495 or g.66493737 T ≻ C, as well as Ins227bp homozygotes earned most money on a mean distance ranging from 1211 to 1230 m. Heterozygotes earned most money on races over 1690 to 1709 m. The BRD for the T/T carriers at both SNP loci and for the SINE-free genotype was 1812 to 1854 m. Other performance parameters were not significantly different between the genotypes, except of the relative success score (RSS). The RSS was significantly slightly better on a distance of ≤ 1300 m for all carriers of the C allele and the Ins227bp compared to homozygous T genotypes and SINE-negative horses (p = 0.037 to 0.046). For distances of more than 1300 m the RSS was not significantly different between genotypes. In silico assessment indicated that the Ins227bp promoter insertion might have generated a CpG island and a few novel putative binding sites for transcription factors. CONCLUSIONS: All three target polymorphisms (Ins227bp, g.66493737 T ≻ C, BIEC2-417495) are suitable markers to assess the ability of non-elite Thoroughbreds to race at short or longer distances. The CpG island generated by Ins227bp may cause training-induced silencing of MSTN expression.

Measurement of tidal volume using respiratory ultrasonic plethysmography in anaesthetized, mechanically ventilated horses.

OBJECTIVE: To compare tidal volume estimations obtained from Respiratory Ultrasonic Plethysmography (RUP) with simultaneous spirometric measurements in anaesthetized, mechanically ventilated horses. STUDY DESIGN: Prospective randomized experimental study. ANIMALS: Five experimental horses. METHODS: Five horses were anaesthetized twice (1 week apart) in random order in lateral and in dorsal recumbency. Nine ventilation modes (treatments) were scheduled in random order (each lasting 4 minutes) applying combinations of different tidal volumes (8, 10, 12 mL kg(-1)) and positive end-expiratory pressures (PEEP) (0, 10, 20 cm H(2)O). Baseline ventilation mode (tidal volume=15 mL kg(-1), PEEP=0 cm H(2)O) was applied for 4 minutes between all treatments. Spirometry and RUP data were downloaded to personal computers. Linear regression analyses (RUP versus spirometric tidal volume) were performed using different subsets of data. Additonally RUP was calibrated against spirometry using a regression equation for all RUP signal values (thoracic, abdominal and combined) with all data collectively and also by an individually determined best regression equation (highest R(2)) for each experiment (horse versus recumbency) separately. Agreement between methods was assessed with Bland-Altman analyses. RESULTS: The highest correlation of RUP and spirometric tidal volume (R(2)=0.81) was found with the combined RUP signal in horses in lateral recumbency and ventilated without PEEP. The bias ±2 SD was 0±2.66 L when RUP was calibrated for collective data, but decreased to 0±0.87 L when RUP was calibrated with individual data. CONCLUSIONS AND CLINICAL RELEVANCE: A possible use of RUP for tidal volume measurement during IPPV needs individual calibration to obtain limits of agreement within ±20%.

In vitro validation of a new respiratory ultrasonic plethysmograph.

OBJECTIVE: The in-vitro validation of a novel Respiratory Ultrasonic Plethysmography (RUP) system designed to detect circumference changes of rib cage and abdominal compartments in large and small animals. STUDY DESIGN: Experimental in vitro study. METHODS: The experimental system includes two compliant fluid-filled rubber tubes functioning as ultrasonic waveguides. Each has an ultrasonic transmitter and a detector at the opposing ends. Sensor length can be individually adapted in the range of 0.15-2 m. Data are downloaded to a computer at a sampling rate of 10 or 100 Hz. Measurements have a resolution of 0.3 mm. Baseline stability, linearity and repeatability were investigated with dedicated experiments. The base line drift was tested measuring a fixed distance for 2 hours continuously and then 18 hours later. A hand-operated horse thorax dummy (elliptically shaped, circumference 1.73 m) was used to compare waveforms of RUP with a respiratory inductive plethysmograph (RIP). The electromagnetic interference was tested by approaching metallic objects. RESULTS: Baseline drift and repeatability (10 repeated steps of 1.6% and 6.6% elongations and contractions) were within ± 0.3 mm. The response of the system for tube stretching up to 11% of total length was linear with a coefficient of determination for linearity of 0.998. In contrast to RIP, electromagnetic interference could not be observed with RUP. CONCLUSIONS AND CLINICAL RELEVANCE: The low baseline drift and the lack of electromagnetic interference favours the use of RUP compared to an RIP device when studying the breathing pattern and end expiratory lung volume changes in conscious and anaesthetized animals.

Emergence of West Nile virus lineage 2 in Europe: Characteristics of the first seven cases of West Nile neuroinvasive disease in horses in Austria.

We report details of the first seven equine cases of confirmed West Nile neuroinvasive disease in Austria. The cases presented during summer and autumn of 2016 (n = 2), 2017 (n = 3) and 2018 (n = 2). All horses showed gait abnormalities and 6 of 7 horses exhibited fasciculations and/or tremors, and we provide video recordings of these. Three horses also showed cranial nerve involvement. Following rapid improvement, three horses were discharged. Four horses were euthanized due to the severity of clinical signs and subjected to neuropathological examination. West Nile virus (WNV) lineage 2 nucleic acid was detected in 5 of 7 horses, and WNV-specific neutralizing antibodies in all 7 horses. In addition, serologic evidence of WNV infection was found in two out of fourteen in-contact horses. Horses may be considered a sentinel species for human WNV infections, integrating human and veterinary medicine and thus contributing to the one health concept.

Effects of Hogweed (Heracleum persicum) Powder, Flavophospholipol, and Probiotics as Feed Supplements on the Performance, Carcass and Blood Characteristics, Intestinal Microflora, and Immune Response in Broilers.

The effect of different levels of hogweed powder (HP; Heracleum persicum), flavophospholipol (antibiotic), and probiotics in diet on the performance, carcass quality, blood biochemical parameters, immunity, and intestinal flora of broiler chickens was investigated. In total, 270-day-old male broilers were randomly assigned to six treatment groups as follows: control basal-diet and diet supplemented with flavophospholipol, probiotics, or 0.25, 0.5, and 0.75% HP. Birds in each group were divided into three subgroups with 15 chicks each. Results indicated that the treatment groups did not vary with respect to feed intake (FI), whereas those supplemented with the antibiotic or 0.5% HP showed significantly higher body weight gain (BWG) and improved feed conversion ratio (FCR). Carcass characteristics did not vary among treatments, with the exception of abdominal fat percentage, which was the lowest in broilers fed 0.5% and 0.75% HP. Supplementation of 0.5% and 0.75% HP decreased plasma cholesterol and triglyceride levels. Furthermore, dietary HP significantly reduced serum low density lipoprotein (LDL) levels compared to that in the other groups. Antibody titers against Newcastle disease vaccine were not markedly affected by the treatments, whereas titers against avian influenza vaccine were significantly higher in probiotic- and 0.75% HP-supplemented groups. Antibody production against sheep red blood cells (SRBC) and IgM and IgG levels were not significantly different among groups. The ileum Lactobacillus counts in broilers fed 0.5% or 0.75% HP were significantly higher than those in the other treatment groups, whereas Escherichia coli counts in all treatments were significantly lower than that in the control. Therefore, our observations indicated that HP positively affected the gut microbiota and enhanced feed digestion. In conclusion, supplementation of 0.50-0.75% HP in broiler diet during the entire rearing period improved BWG and decreased abdominal fat deposition.

Effects of Dietary Supplementation of L-Carnitine and Excess Lysine-Methionine on Growth Performance, Carcass Characteristics, and Immunity Markers of Broiler Chicken.

L-carnitine as well as lysine and methionine are amino acids of important nutritional and nutraceutical interest and are used in nutritional strategies as diet supplements to improve feed quality characteristics in animals and broiler chicken in particular. This study investigated the effect of different levels of L-carnitine and extra levels of lysine-methionine on growth performance, carcass characteristics, and some immune system markers. Two hundred seventy male Ross 308 broilers were a fed control diet (C) and eight different diets supplemented with an excess of amino acids. In the experimental diets, identified as D1, D2, D3, D4, D5, D6, D7, and D8, extra L-carnitine, lysine, and methionine were added in excess with respect to the American National Research Council (NRC) recommendations: L-carnitine equal to NRC (D1); control diet supplemented with lysine at 30% in excess of NRC, methionine at 30% in excess of NRC, and L-carnitine equal to NRC (D2); control diet supplemented with lysine equal to NRC, methionine equal to NRC, and L-carnitine at 15% in excess of NRC (D3); control diet supplemented control diet supplemented with lysine at 15% in excess of NRC, methionine at 15% in excess of NRC, and L-carnitine at 15% in excess of NRC (D4); control diet supplemented lysine at 30% in excess of NRC, methionine at 30% in excess of NRC, and L-carnitine at 15% in excess of NRC (D5); control diet supplemented with lysine equal to NRC recommendations, methionine equal to NRC recommendations, and L-carnitine at 75% in excess of NRC (D6); control diet supplemented with lysine at 15% in excess of NRC, methionine at 15% in excess of NRC, and L-carnitine at 75% in excess of NRC (D7); and control diet supplemented with lysine at 30% in excess of NRC, methionine at 30% in excess of NRC, and L-carnitine at 75% in excess of NRC (D8). During the starter and growth phases, feed intake was not affected by dietary treatment (p > 0.05). By contrast, body weight and FCR were both affected (p < 0.001) during the starter period. During the finisher phase, feed consumption was affected (p < 0.05) by dietary treatment. Feed intake of broilers fed on C, D3, D6, and D7 were statistically similar (p > 0.05) (1851.90, 1862.00, 1945.10, and 1872.80 g/pen/day, respectively) and were higher (p < 0.05) than 1564.40 g/pen/day (D5). With the exception of drumsticks, neck, back thoracic vertebrae, and proventriculus weights, economical carcass segments were not affected (p > 0.05) by the dietary supplementation of amino acids. Duodenum and ileum weights and lengths decreased with amino acid supplementation (p < 0.05). IgT and IgG titers against Sheep Red Blood Cells (SRBC) for both primary and secondary responses were not affected by dietary treatments (p > 0.05). Dietary amino acids supplementation did not affect IgM titer after the secondary challenge (p > 0.05) and had a significant effect (p < 0.05) on serum antibody titers in broilers vaccinated against Newcastle disease (NCD) and Gumboro 's disease at the 27th and 30th days, respectively.

Effects of cadmium chloride inhalation on airflow limitation to histamine, carbachol and adenosine 5'-monophosphate assessed by barometric whole body plethysmography in healthy dogs.

The effects of pharmacological bronchoprovocation on airflow patterns and surrogate respiratory parameters assessed by barometric whole body plethysmography (BWBP) were investigated in healthy dogs, previously exposed to cadmium chloride inhalation. BWBP-derived respiratory variables were calculated (1) at baseline and (2) following nebulisation of increasing concentrations of histamine, carbachol and adenosine 5'-monophosphate (AMP) until enhanced pause (PENH) increased to 300% of baseline (PCPENH300). Bronchoalveolar lavage fluid (BALF) cytology before (BCC) and after (ACC) cadmium chloride inhalation revealed cadmium-induced airway inflammation. Neutrophils increased from 6.7 +/- 7.3% (728 +/- 104/microL) BCC to 77.8 +/- 8.6% (3255 +/- 1407/microL) ACC. PCPENH300 for all three agonists significantly decreased ACC (means+/-SD) as follows: PCPENH300(histamine) 0.72 +/- 0.28 mg/mL BCC, and 0.35 +/- 0.31 mg/mL ACC (P<0.02); PCPENH300(carbachol) 0.34 +/- 0.16 mg/mL BCC, and 0.064 +/- 0.032 mg/mL ACC (P<0.02); PCPENH300(AMP) 1000 mg/mL BCC, and 415 +/- 398 mg/mL ACC (P<0.03). The only clinical sign shown was coughing. It was concluded that airway hyperresponsiveness after induced airway inflammation can be determined by BWBP in conscious small sized dogs. BWBP could be a suitable technique to study the respiratory effects of urban environmental pollution in pets.

Effects of feeding sunflower oil or seal blubber oil to horses with recurrent airway obstruction.

A crossover feeding trial was performed with 9 horses suffering from recurrent airway obstruction (RAO). The study aimed to determine whether ingestion of sunflower oil (SFO), rich in linoleic acid, or seal blubber oil (SBO), a source of long-chain omega-3 polyunsaturated fatty acids (LC n-3 PUFAs), changes the fatty acid (FA) ratios in plasma and leukocyte membrane phospholipids (PLs) or the leukocyte numbers or proportions of cell types in the airways. We also investigated diet-related changes in respiratory rate, maximum change in pleural pressure (deltaPpl(max)), dynamic compliance (C(dyn)), and pulmonary resistance (RL). Each animal was fed hay and oats supplemented with 320 mg/kg body weight (BW) of either SFO or SBO for 10 wk. Before and after the feeding periods, we performed FA analyses, cytologic testing of the pulmonary epithelial lining fluid (PELF), clinical scoring, and pulmonary function testing. The results demonstrated that supplementary FAs were readily ingested and incorporated into leukocyte cell membranes. The n-6:n-3 FA ratios in plasma and leukocyte PLs were reduced after SBO supplementation, as were the PELF leukocyte counts (P < 0.05). On the other hand, pulmonary function and clinical signs were not markedly changed by the different dietary FAs. These results indicate a possible influence of dietary n-3 PUFAs on the pulmonary inflammation of horses with RAO. Further studies are warranted to address effects on inflammatory mediators and clinical outcome.

Effects of dietary inclusion level of a mixture of probiotic cultures and enzymes on broiler chickens immunity response.

The current study was conducted to analyse the effect of a mixture of probiotic cultures and enzymes (Probio Enzyme, XVET GmbH, Germany) on the immune response and weight of central lymphoid organs and liver in broilers. A total of 270 male chickens were randomly divided into nine groups, with three replicates of 10 birds each. Treatment groups were fed for either 22 or 42 days with different levels of Probio Enzyme 250, 500, 750 and 1000 g/ton, whereas the control group fed a basal diet without Probio Enzyme. To analyse the effects of dietary supplementation on broilers humoral immune response, the antibodies titres for avian influenza (AI) and Newcastle disease virus (NDV) and to the sheep red blood cells (SRBC) challenge were assayed in birds from each experimental unit, along with the assessment of the weight of the main lymphoid organs and liver. The addition of the Probio Enzyme mixture did not significantly affect the titres of the antibodies against AI and NDV at day 42, despite the wide individual variation observed specially on the antibody titres at day 33. Treatments affected the production of IgG after the second challenge with SRBC (P = 0.003), which was transposed to the correspondent total Ig titres (P = 0.044). Conversely, a lower birds' body weight (BW) was found in the majority of treated groups compared to control (P = 0.031). The spleen was the only lymphoid organ showing differences in the absolute and relative weight (P = 0.003 and P = 0.001, respectively). No differences were found in thymus and Bursa of Fabricius weights. In conclusion, broilers treated with Probio Enzyme showed a satisfactory immune response compared with control, despite the wide variation found after the first vaccine challenge against AI. Moreover, the probiotic mixture dose and duration modulated differently the immune response and the spleen weight, unaffecting the central lymphoid organs weight.

Retrospective evaluation of 155 adult equids and 21 foals with tetanus in Western, Northern, and Central Europe (2000-2014). Part 1: Description of history and clinical evolution.

OBJECTIVE: To describe clinical data of hospitalized adult equids and foals with tetanus. DESIGN: Multicenter retrospective study (2000-2014). SETTING: Twenty Western, Northern, and Central European university teaching hospitals and private referral centers. ANIMALS: One hundred fifty-five adult equids (>6 months) and 21 foals (<6 months) with tetanus. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Information on geographic, annual and seasonal data, demographic- and management-related data, clinical history, clinical examination and blood analysis on admission, complications, treatments, and outcomes were described and statistically compared between adults and foals. The described cases were often young horses. In 4 adult horses, tetanus developed despite appropriate vaccination and in 2 foals despite preventive tetanus antitoxin administration at birth. Castration, hoof abscesses, and wounds were the most common entry sites for adults; umbilical cord infections and wounds for foals. Stiffness was the commonest observed initial clinical sign. Blood analyses frequently revealed an inflammatory response, hemoconcentration, muscle damage, azotemia, negative energy balance, liver damage, and electrolyte and acid base disturbances. Common complications or clinical signs developing during hospitalization included dysphagia, dyspnea, recumbency, hyperthermia, seizures, hyperlipemia, gastrointestinal impactions, dysuria, and laryngeal spasms. Cases were supported with wound debridement, antimicrobial treatment, tetanus antitoxin, muscle spasm and seizure control, analgesia, anti-inflammatory drugs, fluid therapy, and nutritional support. Mortality rates were 68.4% in adult horses and 66.7% in foals. Foals differed from adult horses with respect to months of occurrence, signalment, management-related data, potential causative events, clinical signs on admission, blood analysis, complications, and severity grades. CONCLUSIONS: This is the first study that rigorously describes a large population of equids affected by tetanus. The information provided is potentially useful to clinicians for early recognition and case management of tetanus in adult horses and foals. Tetanus affects multiple organ systems, requiring broad supportive and intensive care. Neonatal and adult tetanus in the horse should be considered as distinct syndromes, as in human medicine.

Retrospective evaluation of 155 adult equids and 21 foals with tetanus from Western, Northern, and Central Europe (2000-2014). Part 2: Prognostic assessment.

OBJECTIVE: To identify prognostic variables for adult equids and foals with tetanus. DESIGN: Multicenter retrospective study (2000-2014). SETTING: Twenty Western, Northern, and Central European university teaching hospitals and private referral centers. ANIMALS: One hundred fifty-five adult equids and 21 foals with tetanus. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Variables from history and clinical examination were statistically compared between survivors and nonsurvivors (adults: 49 survivors, 85 nonsurvivors; foals: 7 survivors, 10 nonsurvivors). Cases euthanized for financial reasons were excluded. Mortality rates in adults and foals were 68.4% and 66.7%, respectively. Variables associated with survival in adults included: standing, normal intestinal sounds and defecation, voluntarily drinking, eating soft or normal food, lower heart and respiratory rates, high base excess on admission, longer diagnosis time, treatment and hospitalization delay, and mild severity grade. Variables associated with death included: anorexia, dysphagia, dyspnea, low blood potassium concentration on admission, moderate and severe disease grading, development of dysphagia, dyspnea, recumbency and seizures during hospitalization, treatment with glycerol guaiacolate, intravenous fluids, and intravenous glucose solutions. Variables associated with survival in foals included standing on admission, voluntarily eating soft food and drinking, older age, and longer hospitalization delay. Outcome was not different between different tetanus antitoxin (TAT) dosages, although there was a trend of increasing survival rate with increasing TAT dosages. Cases with appropriate vaccination prior to development of tetanus were rare, but had improved outcome and shorter hospitalization. CONCLUSIONS: Prognosis for equine tetanus is poor with similar outcome and prognostic factors in foals and adults. The prognostic assessment of cases with tetanus provides clinicians with new evidence-based information related to patient management. Several prognostic indicators relate to the ability to eat or drink, and more severe clinical signs relate to poor outcome. Increasing intravenous dosages of TAT has no significant effect on outcome, but the positive trend identified may support a recommendation for high intravenous TAT dosages. Further evaluation is warranted.

Effect of Different Dietary Levels of Atorvastatin and L-Carnitine on Performance, Carcass Characteristics and Plasma Constitutes of Broiler Chickens.

The effects of L-carnitine, atorvastatin and their combination on growth and lipid metabolism of broiler chickens is not yet known. Thus, the objective of the present study was to investigate the effects of dietary L-carnitine and atorvastatin on the performance, carcass characteristics and blood parameters in broilers. Different dietary levels of L-carnitine (0, 150 and 300 mg/kg, respectively) and atorvastatin (0, 1 and 2 g/kg, respectively) were added to the daily birds' ration. Significant positive effects (P<0.05) on broiler body weight for both L-carnitine and atorvastatin were reported, and this effect became clear starting from the 4th week of rearing period till the slaughter age. Dietary treatments had also significant (P<0.05) positive effects on broilers empty carcass, breast and drumstick weights. Conversely, L-carnitine slightly increased abdominal fat, whereas supplementing atorvastatin slightly reduced it (P<0.05). However, Combining the treatments, resulted in reduction of abdominal fat pad, showing also the best development of breast and drumstick muscles (P<0.05). Moreover, the weight of gizzard, liver and heart were significantly higher in birds treated with the highest doses supplied (P<0.05). Dietary treatments had also influence on blood biochemical parameters of broilers. In overall, our findings suggest that combining dietary L-carnitine and atorvastatin supported birds growth and muscles development reducing the body fat deposition. However, further studies are needed to deeply study the potential effect of statins on meat quality.

Use of the meridian test for the detection of equine herpesvirus type 1 infection in horses with decreased performance.

OBJECTIVE: To evaluate use of the acupuncture meridian test for detection of recent or recently reactivated equine herpesvirus type 1 (EHV-1) infection in horses with decreased performance. DESIGN: Case-control study. ANIMALS: 40 horses. PROCEDURE: Physical and neurologic examinations were performed, and acupuncture points on the bladder meridian were tested for sensitivity reactions in case and control horses. Polymerase chain reaction assays were performed to determine whether EHV-1 or equine herpesvirus type 4 (EHV-4) DNA could be detected in peripheral blood mononuclear cells. Complement fixation (CF) tests for detection of antibodies against EHV-1 and EHV-4 and virus neutralization (VN) tests for detection of antibodies against EHV-1 were performed on paired serum samples obtained 3 weeks apart. RESULTS: There was a significant difference in skin sensitivity in the cervical, sacral, and gluteal regions and flank between case and control horses. By use of the meridian test, all case horses were sensitive to manipulation of all acupuncture points believed to be associated with EHV infections, whereas only a few control horses were sensitive at an occasional point. Equine herpesvirus type 1 or EHV-4 viremia was not detected in any horses. Mean +/- SDVN antibody titers against EHV-1 were not significantly different between the 2 groups. Mean +/- SD CF antibody titers against EHV-1 obtained 3 weeks after the initial samples were higher in case horses than control horses; however, unequivocal seroconversion was not detected. CONCLUSIONS AND CLINICAL RELEVANCE: Results of the meridian test in case horses were associated with sensitivity reactions similar to those detected by physical and neurologic examinations; however, an unequivocal association with EHV-1 or EHV-4 infection was not detected.

Serological evidence of West Nile virus infection in the horse population of northern Serbia.

INTRODUCTION: This study was conducted to evaluate the seroprevalence of West Nile virus (WNV) in the horse population of northern Serbia. Furthermore, it aimed to provide insight and an updated overview on the circulation of this re-emerging pathogen in this part of southeastern Europe. At the time of manuscript preparation, no clinical cases of WNV infection in horses were reported in this area. METHODOLOGY: Between 2007 and 2011, a total of 252 equine serum samples were collected from seven different locations in northern Serbia. The presence of WNV-reactive IgG antibodies was examined by using commercial and in-house ELISAs. Selected ELISA-positive samples were re-tested by a WNV lineage 2 plaque reduction neutralization test 90% (PRNT-90). RESULTS: In 28.6% of the 252 tested equine serum samples antibodies against WNV were detected. The results obtained with the in-house ELISA corresponded to the outcome of the commercial kit in 90% of the samples. All selected WNV antibody ELISA-positive samples were confirmed by PRNT-90 with neutralizing antibody titers of 1:23 to > 1:512. CONCLUSION: This study confirms the circulation of WNV in northern Serbia. No striking regional differences in seroprevalences were identified suggesting WNV circulation also in other parts of Serbia. Distances between wetlands or forests and stud farms do not appear to have an influence on WNV seroprevalence.

Daily variability of forced oscillometry parameters in horses suffering recurrent airway obstruction, a pilot study.

The aim of the study was to analyse the day-to-day variability of the respiratory resistance (R(rs)) and the reactance (X(rs)) in 5 horses in a status of remission of recurrent airway obstruction by forced oscillometry system (FOS). Furthermore, the effects of stabling, outdoor and room temperature and humidity on these parameters were determined. Mean R(rs) at oscillation frequencies 1, 1.5, 2 and 3 Hz were all significantly higher in the morning than in the afternoon, while X(rs) was not significantly affected by time of the day. R(rs) was significantly different on various sampling days at all frequencies, in the morning as well as in the afternoon. X(rs) showed the same significant differences for frequencies greater than 1 Hz. Overall, R(rs) showed a smaller variability than X(rs). Neither outdoor temperature, nor room temperature affected R(rs). This also applied to X(rs) except for the measurements at 2 Hz that were significantly (p = 0.044) affected by outdoor temperature. Turning out these horses at paddock for day and night caused a significant lower mean R(rs) that when kept at stable for all night. While R(rs) showed a very weak positive frequency dependency, X(rs) clearly showed a positive frequency dependence for all horses. Resonance frequency (f(res)) was between 2 and 4 Hz. These studies showed that FOS generated parameters are affected by environmental and management factors and thus only after well-designed standardized procedures FOS could be a useful diagnostic tool for the diagnosis and monitoring of equine respiratory disorders.

Diagnostic value of the neutrophil myeloperoxidase index in horses with systemic inflammation.

The myeloperoxidase index (MPXI) was investigated as a diagnostic indicator of systemic inflammation in a retrospective study using data from 859 hospitalised horses. A reference interval of 8.5-10.4 for the MPXI was established. In horses with systemic inflammatory response syndrome (SIRS), the MPXI was significantly lower than in healthy horses, those with localised inflammation and those with sepsis. The MPXI in horses with sepsis was also significantly lower than in healthy animals and those with localised inflammation. Horses in the SIRS group with leucopenia, white blood cell (WBC) count within the reference interval (WRI) or leucocytosis had significantly lower MPXIs than healthy horses, those with localised inflammation and those with sepsis in the same WBC count subgroups. In horses with sepsis and WBC count WRI, the MPXI was significantly lower than in healthy horses or those with localised inflammation. MPXI is a useful complementary tool to identify horses with systemic inflammation, especially if they have WBC counts WRI.

Effect of different levels of L-carnitine and lysine-methionine on broiler blood parameters / Efecto de suplementación con L-carnitina y lisina-metionina sobre parámetros de sangre de pollos de engorde

Objective. In the present study a completely randomized 3x3 factorial design was used to analyze the effects of different levels of L-Carnitine, lysine(Lys) and methionine (Met) on the blood concentrations of energy, protein and lipid metabolites of male broiler chickens. Materials and methods. A total of 270 newly hatched male broiler chickens (Ross 308) were randomly assigned to 9 groups (ten broilers per replicate and three replicates per treatment). The control group was fed a basal diet, whereas the treatment groups were fed basal diets supplemented with L-Carnitine (0 mg/kg, 75 mg/kg and 150 mg/kg) and lysine-methionine (0, 15 and 30%) for 42 days. On day 42, one bird was randomly chosen per replication, a blood sample was taken and the blood concentrations of glucose (GLU), uric acid (UAc), triglyceride (TG), VLDL, HDL, LDL, total protein (TP), albumin (Alb) and total cholesterol (TC) were analyzed. Results. Dietary L-carnitine supplementation had a significant effect (p<0.05) on uric acid (UAc), HDL, LDL, and total cholesterol (TC). The birds feed L-carnitine plus Lys and Met presented the highest plasmatic UAc level and the lowest plasmatic TC and LDL level. Moreover, L-carnitine significantly reduced total cholesterol (TC) when compared with both the control group and the birds feed Lys and Met without L-carnitine. Conclusions. A diet with 150 mg/kg L-carnitine plus 15% Lys and Met seems to be enough to sustain low plasmatic TC, LDL and HDL concentrations on male broiler.

Objetivo. Se realizó un estudio para determinar el efecto de la suplementación deL-carnitina, lisina (Lys) y metionina (Met) sobre los metabolitos sanguíneos de pollos de engorde. Materiales y métodos. Se utilizaran 270 pollos de la línea Ross 308 de un día de edad y se dividieron en 9 tratamientos en un diseño al azar con arreglo factorial 3x3: tres niveles de L-carnitina (0 mg/kg, 75 mg/kg y 150 mg/kg) y tres de lisina-metionina (0, 15 y 30%) durante 42 días. Cada tratamiento constó de 3 repeticiones con 10 pollos por repetición. El día 42 de edad, se tomaron muestras de sangre de tres aves por tratamiento para cuantificar niveles séricos de glucosa (GLU), ácido úrico (UAc), triglicéridos (TG), VLDL, HDL, LDL, proteínas totales (PT), albúmina (Alb) y colesterol total (TC). Resultados. La suplementación de L-carnitina en la dieta tuvo un efecto significativo (p<0.05) en los niveles de ácido úrico en suero (UAC), HDL, LDL y colesterol total (CT). Las aves alimentadas con L-carnitina más Lys y Met mostraron niveles séricos más altos de UAc y menor TCy LDL. Por otra parte, la L-carnitina redujó significativamente el colesterol total (CT), cuando se comparó con el grupo control y con los pollos alimentados con Lys y Met, sin L-carnitina. Conclusiones. Una dieta con 150 mg/kg de L-carnitina y 15% Lys y Met parece ser suficiente para mantener bajas concentraciones plasmáticas de TC, LDL y HDL en pollos de engorde.

Influence of the collection and oxygenation method on quantitative bacterial composition in bronchoalveolar lavage fluid samples from healthy dogs.

The purpose of the study was to evaluate the effects on quantitative and qualitative microbial content of endoscopic bronchoalveolar lavage fluid (BALF) in healthy dogs using a laryngeal mask airway (LMA). It was hypothesised that oropharyngeal protection might prevent contamination of BALF with oropharyngeal microflora. Ten healthy Beagle dogs were randomly assigned to two groups and underwent endoscopic BAL on two occasions, either via an open unprotected oropharynx with oxygen supply provided via a nasal catheter (NT) or through a sterile LMA. For the second sampling, groups were switched. BALF analysis included quantitative microbial culture, nucleated cell counts and cytology. The mean (+/-SD) number of colony forming units (CFU)/mL found in the BALF using the LMA was 25,610+/-22,943 in the right lung (RL) and 22,510+/-18,779 in the left (LL). With the NT technique, the figures were 21,068+/-19,375 for the RL and 16,060+/-15,523 for the LL, respectively. Nucleated cell counts/microL were 691.0+/-181.6 (RL) and 734.0+/-171.6 (LL) for LMA, and 772.0+/-251.0 (RL) and 748+/-163.2 (LL) for NT. No significant differences were detected either in the number of CFU/mL or in the diversity of bacterial species with the two methods. A significant increase in BALF bacterial counts (with reduced species diversity) was observed on the second compared to the first sampling regardless of the method used. Protection of the oral cavity and oropharynx using an LMA had no significant influence on BALF bacterial counts. The findings suggest that with careful endoscope insertion, the risk of contamination of BALF by resident and transient oropharyngeal microflora can be negligible.

Effects of butorphanol on the withdrawal reflex using threshold, suprathreshold and repeated subthreshold electrical stimuli in conscious horses.

OBJECTIVE: To assess the effects of a single intravenous dose of butorphanol (0.1 mg kg(-1)) on the nociceptive withdrawal reflex (NWR) using threshold, suprathreshold and repeated subthreshold electrical stimuli in conscious horses. STUDY DESIGN: 'Unblinded', prospective experimental study. ANIMALS: Ten adult horses, five geldings and five mares, mean body mass 517 kg (range 487-569 kg). METHODS: The NWR was elicited using single transcutaneous electrical stimulation of the palmar digital nerve. Repeated stimulations were applied to evoke temporal summation. Surface electromyography was performed to record and quantify the responses of the common digital extensor muscle to stimulation and behavioural reactions were scored. Before butorphanol administration and at fixed time points up to 2 hours after injection, baseline threshold intensities for NWR and temporal summation were defined and single suprathreshold stimulations applied. Friedman repeated-measures analysis of variance on ranks and Wilcoxon signed-rank test were used with the Student-Newman-Keul's method applied post-hoc. The level of significance (alpha) was set at 0.05. RESULTS: Butorphanol did not modify either the thresholds for NWR and temporal summation or the reaction scores, but the difference between suprathreshold and threshold reflex amplitudes was reduced when single stimulation was applied. Upon repeated stimulation after butorphanol administration, a significant decrease in the relative amplitude was calculated for both the 30-80 and the 80-200 millisecond intervals after each stimulus, and for the whole post-stimulation interval in the right thoracic limb. In the left thoracic limb a decrease in the relative amplitude was found only in the 30-80 millisecond epoch. CONCLUSION: Butorphanol at 0.1 mg kg(-1) has no direct action on spinal Adelta nociceptive activity but may have some supraspinal effects that reduce the gain of the nociceptive system. CLINICAL RELEVANCE: Butorphanol has minimal effect on sharp immediate Adelta-mediated pain but may alter spinal processing and decrease the delayed sensations of pain.