Francesca Bonelli

Plasma Procalcitonin Concentration in Healthy Horses and Horses Affected by Systemic Inflammatory Response Syndrome

Background The diseases most frequent associated with SIRS in adult horses are those involving the gastrointestinal tract. An early diagnosis should be the goal in the management of horses with SIRS. Objective The objective of this study was to evaluate the plasma procalcitonin (PCT) concentration in healthy and SIRS horses to assess differences between the two groups. Animals Seventy‐eight horses (30 healthy and 48 SIRS). Methods Prospective in vivo multicentric study. Horses were classified as SIRS if at least 2 of the following criteria were met: abnormal leukocyte count or distribution, hyperthermia or hypothermia, tachycardia, tachypnea. Healthy horses showed no clinical or laboratory signs of SIRS. Plasma PCT concentrations were measured with a commercial ELISA assay for equine species. Results were expressed as mean±standard deviation. T‐test for unpaired data was performed between healthy and SIRS group. SIRS group was divided in 4 subgroups and t‐test was performed between healthy versus each subgroup. Results PCT concentrations in healthy and SIRS horses were 18.28 ± 20.32 and 197.0 ± 117.0 pg/mL, respectively. T‐test showed statistical differences between healthy versus SIRS group and between healthy versus all subgroups. Conclusions and Clinical Importance Results showed an increase in PCT concentration in SIRS horses as previously reported in humans and dogs. PCT could be used as a single assay in equine practice for detection of SIRS.

Conjunctival bacterial and fungal flora in clinically normal sheep.

Objectives The aim was to identify conjunctival bacterial and fungal flora in clinically normal sheep. Design Prospective study. Setting Tuscany. Participants 100 eyes from 50 adult Massese female sheep were examined. The sheep included in the study were considered free of anterior ophthalmic abnormalities. Primary and secondary outcome measures Bacteria were identified by morphological assessment, Gram staining, biochemical tests. Identification of filamentous fungi was achieved at the genus level, and Aspergillus species were identified based on keys provided by other authors. Yeast colonies were highlighted, but not identified. Results Positive cultures were obtained from 100/100 eyes for bacteria, and from 86/100 eyes for fungi. A total of 14 types of bacteria and 5 types of fungi were isolated. Yeasts were isolated from 13/100 eyes. The most frequent fungal isolates were saprophytic fungi. Conclusions Conjunctival bacterial and fungal flora of clinically normal eyes were reported in sheep. The positivity obtained for conjunctival bacteria was higher compared to findings in the literature by other authors in the same species (100 per cent v 40 per cent), while our results were in line with a recent work performed on mouflons (Ovis Musimon) with a 100 per cent positivity for bacterial conjunctival fornix. In our survey, Gram-positive species were prevalent, as reported by other authors in different species. Few data are available in the literature regarding conjunctival fungal flora in healthy small ruminants. The prevalence of conjunctival fungal flora in this study was higher than findings reported in mouflons (86 per cent v 45 per cent). Differences in fungal prevalence may be due to different methods of managing herds, though further studies are required to verify this hypothesis. The similarities in bacterial and fungal isolates between sheep and mouflons suggest a genera pattern of conjunctival colonisation by bacteria and fungi.

How swimming affects plasma insulin and glucose concentration in Thoroughbreds: A pilot study

Low intensity exercise increases insulin-stimulated glucose uptake in skeletal muscle and decreases its plasma concentration. In this study, plasma insulin and glucose concentrations were evaluated 5min before and 5, 15, 25, 35, 45 and 60min after an IV bolus of glucose in 12 Thoroughbreds before and after 1 month of submaximal aquatraining exercise, monitored using heart rate and blood lactate. Plasma glucose concentrations were evaluated using a colorimetric enzymatic method, and plasma insulin concentrations with a solid-phase radioimmunoassay method. Pre-training plasma glucose concentrations at 15, 25 and 35min, area under the glucose curve and peak glucose concentration were significantly higher than post-training values (P<0.05). Baseline pre-training plasma insulin concentrations were significantly lower than in the post-training period, and plasma insulin was significantly higher at 45 and 60min in the pre-training period than the post-training period. These results indicate that aquatraining could improve insulin-glucose metabolism in horses.

Kinetics of plasma procalcitonin, soluble CD14, CCL2 and IL-10 after a sublethal infusion of lipopolysaccharide in horses

Endotoxemia represents a significant clinical and economic problem for the equine industry. This study assesses the kinetics of soluble CD14 (sCD14), chemokine (CC motif) ligand 2 (CCL2), interleukin 10 (IL-10) and plasma procalcitonin (PCT) in healthy horses after the intravenous infusion of lipopolysaccharide (LPS). The aim was to contribute to the basic understanding of the equine species-specific kinetics of these molecules in response to LPS exposure, which could support further findings in clinical studies and identify valuable inflammatory biomarkers for equine practice. Eleven healthy horses were involved in this experimental in vivo study. Horses were classified as healthy before the LPS infusion. After the pre-infusion blood collection (T0), all horses received an infusion of E. coli endotoxin (30ng/kg over 30min). Data and samples were collected 1h (T1), 2 (T2), 3 (T3) and 24h (T24) after infusion. Plasma sCD14, CCL2 and IL-10 were evaluated with a fluorescent bead-based assay, while PCT was evaluated with an equine PCT ELISA assay. A one-way ANOVA test was performed between each blood-sampling time for PCT, sCD14 and IL-10, and a Friedman test was performed for CCL2. Plasma PCT, IL-10 and CCL2 concentrations increased statistically significantly at T1, T2 and T3 compared to T0. No statistically significant differences were found between plasma IL-10 and CCL2 concentrations between T0 vs T24, although plasma PCT values remained high 24h after LPS infusion. Plasma sCD14 concentration showed no statistically significant differences for any of sampling times. Our results demonstrate that LPS injection into healthy horses results in PCT, CCL2 and IL-10 increases in plasma without an increase in sCD14. The increases in PCT, CCL2 and IL-10 are related to the inflammatory response induced by circulating lipopolysaccharide.

Evaluation of a Smartphone Electrocardiograph in Healthy Horses: Comparison With Standard Base-apex Electrocardiography

ABSTRACT A good diagnostic accuracy of smartphone‐based electrocardiography (ECG) in the evaluation of heart rate, heart rhythm, and ECG measurements has been reported in humans, dogs, and cats. The aim of this study was to assess the feasibility of smartphone‐based electrocardiography in horses. Fifty healthy adult horses were enrolled. Standard base‐apex ECG and smartphone ECG were simultaneously recorded in each horse. All ECGs were reviewed by one blinded operator, who judged whether tracings were acceptable for interpretation and performed electrocardiographic measurements. Agreement between smartphone and standard base‐apex ECG in the analysis of tracings was evaluated. Smartphone ECG tracings were interpretable in 48 of 50 (96%) cases. A perfect agreement between smartphone and standard ECG tracings was found in the assessment of heart rate. Heart rate automatically measured by the smartphone application was not reliable. In terms of electrocardiographic waves and interval duration, minimal differences of no clinical value were found between smartphone and standard ECG. Agreement was found for QRS complex polarity evaluation, but not for P wave polarity. Baseline artifacts were rare but significantly higher in the smartphone ECG tracings than those in standard ECGs. The smartphone ECG can record single‐lead ECG tracings of an adequate quality for interpretation in horses. The smartphone ECG could represent an additional tool for the electrocardiographic evaluation of horses but is not a substitute for the standard base‐apex ECG. HighlightsThe viability of smartphone electrocardiography (ECG) in horses has been assessed.The smartphone could record good‐quality ECG tracings in 96% of horses.Reliable heart rate was obtained when manually measured on digitized tracings.The smartphone device was reliable for measurement of ECG wave and intervals.The smartphone ECG can be useful for electrocardiographic screening in horses.

Renal Measures in Healthy Italian Trotter Foals and Correlation Between Renal and Biometric Measures: Preliminary Study

Abstract The aim of this study was to evaluate ultrasonographic renal measures in healthy foals aged 1–6 weeks and to verify the correlation between biometric measures to ultrasonographic renal ones. A total of nine Italian trotter foals born in the same stud farm and underwent similar management conditions were enrolled. Inclusion criteria were normal gestation time, unassisted delivery, and normal physical examination at all evaluation times. Length and height of both kidneys were measured by ultrasound weekly from 1 to 6 weeks of life, along with the thoracic and the middle third of the metacarpal area circumferences. Data were expressed as mean and standard deviation, and distribution was evaluated. One‐way analysis of variance (ANOVA) was applied to verify differences related to time. The Pearson correlation test was carried out to evaluate the linearity between time versus all the parameters measured. Students t test was used to verify differences in ultrasound measures between right and left kidney at all recorded times. The Pearson test was applied to a mean‐variance matrix to verify the correlation between each biometrical versus all renal measures. Significance level was set at P < .05. One‐way ANOVA showed differences in biometric and renal measures related to time. Correlation test revealed a linear growth. Differences in ultrasound renal measures between right and left kidney were obtained. Correlation was found between biometrical parameters versus kidney measures. Renal measures and differences between left and right kidneys were in line with literature. Correlation test revealed a linear growth. Renal growth is correlated with age and biometric measures.

Validation of a Paraoxon-based method for measurement of Paraoxonase (PON-1) activity and establishment of RI in horses

Paraoxonase-1 (PON-1) is an anti-oxidant compound considered as negative acute phase protein in animals ( Rossi et al., 2013 ) and people ( Novak et al., 2010 ). The paraoxon-based method for measurement of PON-1 in equine serum has not yet been validated. The aim of this study is to validate a paraoxon-based method to measure PON-1 and to establish reference intervals (RIs) in healthy horses and foals. 120 horses (40 geldings, 40 stallions, 40 mares; median age: 11 years; 57 Warmbloods, 46 Trotters) and 55 foals (27 females, 28 males; median age: 47 days; 22 Warmbloods, 31 Trotters) considered healthy after physical examination and biochemistry were examined. Horses were grouped by breed: Thoroughbreds, Trotters, Warmbloods, Draft horses and Ponies. Serum PON-1 was measured with an automated spectrophotometer and an enzymatic method validated in other species ( Giordano et al., 2013). After the analytical validation (precision, accuracy, interference studies), RIs were determined using the Reference Value Advisor software, according to ASCVP guidelines ( Friedrichs et al., 2012 ). The possible gender-, age- and breed-related differences were statistically investigated. The paraoxon-based method was precise (CVs <4.0%) and accurate (P<0.001 in linearity under dilution and spike-recovery testing) but is affected by interference from mild bilirubinemia, severe lipemia or hemoglobinemia. The RIs recorded in the whole population was 38.1-80.8 U/mL. According to the Harris and Boyd test, separate RIs are recommended only for adult females and for Warmblood and Trotter adults (Figure 1). This study demonstrated that analytical performances of the paraoxon-based method for measurement of PON-1 in horses are acceptable. PON-1 is lower in horses than in other species. If future studies will demonstrate that oxidative stress induces a significant decrease of PON-1, this results will be useful to correctly classify healthy and sick horses; PON-1 could be used, as in human medicine, as a marker of oxidative stress.