PON1 enzyme activity assays for serum and heparinized plasma in horses and stability evaluation of the enzyme activity over different freeze-thaw cycles and mimic transportation.

Consistent information and standardization procedures regarding the time of storage for frozen samples and the effects of storage time on enzyme activity are still missing in the literature. Thus, we evaluated the effects of different storage temperatures (-20 °C and - 80 °C), three repetitive freeze/thaw cycles, and 24-h mimic transportation on the activities of PON1 (paraoxonase and arylesterase), enzymes involved in the protection and detoxification processes of reactive molecules. PON1 enzymes' activity was validated on serum and heparinized plasma in horses. The results revealed that conditions and time of storage of blood samples for PON1 analyses altered the activities of both enzymes in both sample types, evidencing that these conditions can lead to protein degradation or general alteration. Specifically, paraoxonase and arylesterase activities significantly decreased among storage temperatures, with major effects detected at -20 °C. The repeated freeze/thaw cycles at -20 °C and 24-h mimic transport conditions also generated an expected degradation of the arylesterase in both serum and heparinized plasma while freeze/thaw cycles at -80 °C caused an increase of both arylesterase and paraoxonase activities on both sample types. In general, similar enzyme responses were detected between serum and heparinized plasma.

Transcutaneous Auricular Vagal Nerve Stimulation in Healthy Non-Sedated Horses: A Feasibility Study.

This study aimed to evaluate the feasibility of transcutaneous auricular vagal nerve stimulation (tAVNS) in healthy horses and its effect on heart rate variability (HRV). The study comprised three phases: the selection of mares, their acclimatization to the tAVNS, and the stimulation phase. Stimulation was performed with two electrodes positioned on the right pinna. The settings were 0.5 mA, 250 µs, and 25 Hz for pulse amplitude, pulse width, and pulse frequency, respectively. HRV was analysed before (B1), during (T), and after (B2) the tAVNS. From the 44 mares initially included, only 7 completed the three phases. In these mares, the heart rate (HR) was significantly lower, and frequency domain parameters showed an increased parasympathetic tone in B2 compared with B1. However, in 3/7 mares, the HR was significantly higher during T compared with B1 and B2, compatible with a decreased parasympathetic tone, while in 4/7 mares, the HR was significantly lower and the parasympathetic nervous system index was significantly higher during T and B2 compared with B1. The tAVNS is an economical and easy procedure to perform and has the potential to stimulate vagal activity; however, it was poorly tolerated in the mares included in this study.

Evaluation of veterinary students' suture performance according to three different instructional modalities.

Background: Higher education attempts to ameliorate the learning experience through match between learning subjects and students' learning styles. Aim: This study evaluates the efficacy of three different instructional modalities aimed at teaching veterinary students how to make simple suture knots. Methods: A cohort of 43 fourth-year students were split into 3 groups and provided with different instructional modalities: presentation with pictures and descriptions, hard copy text, and muted video. The student's surgical simulation performance was evaluated. Then, they answered a 23 question-survey, Fleming VARK questionnaire based, investigating their learning profile. Kruskal Wallis test evaluated different instructional modalities effect on student's performance. The chi-square test assessed differences between instructional modalities and learning profiles, profile self evaluation, and training session comprehension. Results: Students showed auditory unimodal VARK profile (16/43), did not know their learning profile (26/43), and favored personalized teaching strategies (43/43). No differences were found for: surgical performance, except for forceps handling; and between instructional modalities: either for learning profiles (p-value = 0.43), or profile self evaluation (p-value = 0.42). Differences were found between instructional modalities and training session comprehension. As limitations, auditory instructional modalities, participants' age, and gender were not recorded or evaluated. Conclusion: Our study provides feedback on modern teaching modalities in which students play a key role. Participants showed a variety of learning profiles although displaying no significant performance differences.

The application of a single session of capacitive resistive electric transfer 24 h before exercise modifies the accelerometric pattern in standardbred racing trotters.

BACKGROUND: It has been reported that capacitive resistive electric transfer (CRET) increases blood circulation, hemoglobin oxygenation and temperature in muscles. The attributed benefits of these changes have been linked to improved athletic performance, enhanced muscle flexibility and fastening recovery from exercise-induced fatigue. For all of this, the present research aims to investigate whether the application of CRET 24 h before exercise affects the accelerometric pattern in horses during exercise. Six sound Standardbred trotters were subjected to a CRET session of 40 min of duration, applied on both sides of the neck, back and croup, 24 h before a training session. Training sessions consisted of a warming-up (WU) for 6400 m and a training bout (TB) at their maximal training speed for 1600 m. The same protocol was followed for the device off (sham protocol), also applied 24 h before the training session. CRET and sham experiments were separated by one week, the order of application of both was randomly defined for each individual and drivers were blinded for the duration of the experiment. During the training sessions, horses wore an accelerometer fixed at the sternal level. Speed, stride frequency (SF), length (SL), regularity and symmetry and accelerometric activities were measured during WU and TB. RESULTS: CRET increased speed, mediolateral and total accelerometric activities during WU and speed, SL, dorsoventral, longitudinal and total accelerometric activities during TB, but stride regularity and symmetry decreased. CONCLUSION: The application of CRET 24 h before exercise increased speed and accelerometric activities, results that highlight the need to evaluate the interaction between CRET and training in order to develop new methods to limit fatigue. However, the decrease in stride regularity and symmetry after CRET application could be negative effects, which could be attributed to the increased speed.

Factors related to longevity and mortality of dogs in Italy.

Besides its translational value, an improved understanding of dog longevity and mortality is necessary to guide health management decisions, breed selection, and improve dog welfare. In order to analyse the lifespan of dogs in Italy, identify the most common causes of death, and evaluate possible risk factors, anonymised medical records were collected from 9 veterinary teaching hospitals and 2 public health institutions. Data regarding breed, sex, neuter status, age, diagnosis, and mechanism of death were retrieved. Cause of death (COD) was classified by pathophysiologic process (PP) and organ system (OS). Of the 4957 dogs that died between 2004 and 2020 included in the study, 2920 (59.0%) were purebred, 2293 (46.2%) were female, 3005 (60.6%) were intact, 2883 (58.2%) were euthanised. Overall median longevity was 10.0 years. Median longevity was significantly longer for crossbreds, females, neutered dogs, and small-sized breeds. The breeds with the highest median age at death were the Yorkshire terrier, English cocker spaniel, West Highland white terrier, Italian volpino, and Shih Tzu, whilst the American bulldog, English bulldog, American pit bull terrier, Bernese mountain dog and the Maremma and the Abruzzes sheepdog had the lowest median age at death. The most frequent COD by PP was neoplasia (34.0%), which occurred more frequently in large breeds, namely German shepherd, Labrador retriever and Boxer. Degenerative diseases mostly affected small-sized dogs like Miniature pinscher and Dachshund. Regarding the OS involved, diseases of the renal/urinary system were most frequently responsible for COD (15.0%), prevalently degenerative and inflammatory/infectious. Substantial variation in median longevity according to causes of death by PP and OS was observed. These data are relevant for breeders, veterinary practitioners, and owners, to assist breed selection, facilitate early diagnosis, guide choice when purchasing a purebred dog and making health management decisions, and ultimately improve dog welfare.

Pattern of prescriptions and prudent use of antimicrobial in horse practice at a Veterinary Teaching Hospital.

Antimicrobial prescriptions of the University of Pisa and their compliance with prudent use recommendations were investigated over 11 years (2011-2021). At least one antimicrobial was always prescribed in surgical prophylaxis for the suture of wounds and in 33% of horses with signs of disease of a body system. Antimicrobials were administered in monotherapy (48%) in fixed dose combinations (21%) and empirical combinations (31%). Antimicrobials were mostly (63%) administered by parenteral route, while oral and topical antimicrobials accounted for 14% and 23% of prescriptions, respectively. Gentamicin, benzylpenicillin and ceftiofur were the most prescribed antimicrobials; aminoglycosides, penicillins and cephalosporins were the most common class of antimicrobial prescribed. Protected antimicrobials (WHO HPCIA and rifampicin) represented 24% of antimicrobial dispensations. The pattern of classes of antimicrobial used by body system was broad and included up to eight different pharmaceutical classes. The heterogeneity of antimicrobial use was confirmed by the estimate of the prescription diversity index. Antimicrobial prescriptions were in compliance with prudent use recommendations in terms of availability of diagnosis, respect of the dose range and duration of treatment On the contrary, principles of appropriate antimicrobial use have only been partially observed in relation to off-label use, use of antimicrobials in empirical combination, use of antimicrobial susceptibility tests and use of protected antimicrobials, suggesting that additional interventions are required to improve the responsible use of antimicrobials use in our equine practice.

Evaluation of a smartphone electrocardiograph in healthy foals and comparison to standard base-apex electrocardiography.

Smartphone-based technology for ECG recording has recently spread as a complementary tool for electrocardiographic screening and monitoring in adult horses and in other animal species. The present study aimed to assess the feasibility and accuracy of a smartphone-based ECG in healthy foals. This was a prospective observational study (authorization n. 45,865/2016) including 22 foals aged less than 21 days. A reference standard base-apex ECG (rECG) was acquired, and a smartphone ECG (sECG) was recorded immediately after by using a smartphone-based single lead electrocardiograph. All ECG tracings were evaluated in a blind fashion by a single board-certified cardiologist, who judged whether the tracings were acceptable for interpretation and performed ECG measurements and diagnosis. The Spearman correlation coefficient, the Cohen's k test and the Bland-Altman test were used to assess the agreement between sECG and rECG. All sECG tracings were acceptable for interpretation. All foals showed sinus rhythm on both rRCG and sECG tracings, with perfect agreement in heart rate classification (κ = 0.87; p < 0.001). No clinically relevant differences were found in the assessment of waves and intervals duration. Concerning P wave and QRS complex polarity, the percentage of agreement between rECG and sECG was 78% and 83%, respectively. About ECG tracing quality, rECG and sECG showed a substantial agreement (κ = 0.624; p < 0.001). In conclusion, the smartphone-based ECG device tested in the present study recorded good quality single-lead ECG tracings in foals, reliable for heart rate and ECG measurements, but different polarity of P waves and QRS complexes was found in some foals in comparison to rECG.

Hematology and clinical chemistry in mule foals from birth to two months of age: A preliminary study.

In horses and donkeys, age-related changes in hematological and biochemical parameters preclude the use of normal values of adults in the evaluation of foals. This study aimed to obtain data on hematological and biochemical parameters of mule foals from birth up to the second month of life and to assess age-related changes in order to determine if dedicated reference ranges are required in younger animals. Blood samples from seven healthy mule foals were obtained at birth before colostrum consumption, 24 h, 48 h of life, and then weekly until the second month of life. Results were expressed as mean and standard deviation or median, minimum, and maximum values if showing non-gaussian distribution. Kruskal-Walls and Dunn tests were used to verify the differences among sampling times. Significance was set at P < 0.05. Red blood cell count, packed cell volume and hemoglobin decreased from 24 h to one week of age. Mean corpuscular volume and mean corpuscular hemoglobin decreased over the first month. White blood cells increased from birth to seven days of life. Aspartate amino transferase increased while alkaline phosphatase decreased in the first week of life. Urea, creatinine, and lactate decreased, while glucose concentrations increased at 24 h. Ionized calcium and magnesium and total sodium and potassium showed no changes. In mule foals, several laboratory parameters may be the same or intermediate, lower or higher than in equine or donkey foals, but also compared to all other adult species. The preliminary results suggest that for mule foals, age influences hematological and biochemical parameters.

Procalcitonin and carbonylated protein concentrations in equine synovial fluid.

Early diagnosis of joint diseases is fundamental for prompt and appropriate management, particularly in septic arthritis. Procalcitonin (PCT) and protein carbonylated content (PCC) have been investigated in both human and veterinary medicine. An increase in PCT has been shown in infectious bacterial diseases, while higher levels of PCC have been shown in inflammatory pathologies characterized by oxidative damage. This study evaluated PCT and PCC in plasma and synovial fluid (SF), in healthy and pathological equine joints, affected by different types of arthropathy. Twenty-nine joints were evaluated and underwent orthopedic, radiographic, ultrasonographic and SF evaluation. The joints were divided in three groups: healthy, septic, and non-septic arthritis. PCT and PCC were measured in horse plasma and SF. Data distribution was evaluated and results were expressed as median, quartile values. Statistical differences in SF values among groups and correlations were assessed between plasma and SF of both PCT and PCC. The groups of joint disease included: 8/29 healthy, 13/29 non-septic and 8/29 septic. Significant differences were obtained for SF PCC and plasma PCT between healthy and septic joints, while no differences were found for plasma PCC and SF PCT. A positive correlation was found between plasma and SF PCT. To the best of our knowledge, this is the first study reporting PCT in equine SF. SF PCC could be a useful biomarker to differentiate between septic and healthy joints.

Case-Control Study: Endogenous Procalcitonin and Protein Carbonylated Content as a Potential Biomarker of Subclinical Mastitis in Dairy Cows.

Procalcitonin (PCT) and protein carbonylated content (PCC) are promising biomarkers for bacterial infection and inflammation in veterinary medicine. This study examined plasma PCT and PCC levels in healthy cows (H) and cows with subclinical mastitis (SCM). A total of 130 cows (65 H and 65 SCM) were included in this study. Blood samples were collected, and plasma was frozen at -80 °C. PCT levels were determined using a bovine procalcitonin ELISA kit, while PCC was measured following the methodology of Levine et al. Statistical analysis revealed a significant difference in PCT levels between H (75.4 pg/mL) and SCM (107.3 pg/mL) cows (p < 0.001) and significantly lower concentrations of PCC in the SCM group (H: 0.102 nmol/mL/mg, SCM: 0.046 nmol/mL/mg; p < 0.001). The PCT cut-off value for distinguishing healthy and subclinical mastitis animals was >89.8 pg/mL (AUC 0.695), with a sensitivity of 66.2% and specificity of 69.2%. PCT showed potential value as a diagnostic tool to help in decision making for subclinical mastitis cases, while PCC requires further studies to investigate the trend of this biomarker during localized pathology.

Apgar Score, Clinical, Hemato-Biochemical, and Venous Blood Gas Parameters in a Cohort of Newborn Mule Foals: Preliminary Data.

The study aimed to assess Apgar scores and collect data on clinical, hemato-biochemical, and venous blood gas parameters in seven mules during their first 48 hours of life. Apgar scores, heart rate (HR), respiratory rate (RR), body temperature (BT), body weight (BW), time to achieve sternal recumbency, stand, nurse, and pass meconium were recorded. Blood samples were collected at birth (T0), 6 (T6), 12 (T12), 24 (T24), 48 (T48) hours of life. Data distribution was verified and differences in laboratory parameters over time assessed. One foal had an Apgar score of 6/8, thus excluded from the statistical analysis. Three foals had an Apgar score of 7/8, three had a score of 8/8. All foals showed suckling reflex within 20 minutes and raised the head within 1 minute; six foals reached sternal recumbency within 2 minutes. Times to stand and nurse were 38.7 ± 13.4 and 72.3 ± 30.5 minutes, respectively. Five foals passed meconium spontaneously in 416.3 ± 401.8 minutes. The mean HR, RR, and BT values were 92 ± 16.1 bpm, 50.2 ± 5.9 bpm, and 37.6 ± 0.3°C, respectively. Creatinine and lactate decreased from birth to 48 hours. Blood glucose increased from 12 hours. Mg increased from 12 to 24 hours. Compared to horses and donkeys, mules showed an intermediate BW, shorter time to stand and nurse, longer time to pass meconium, intermediate HR and lower RR. Compared to donkey mules showed lower BT. Other dissimilarities from the parental species were found in laboratory parameters. Our findings emphasize the need for reference ranges for a comprehensive evaluation of newborn mules.

Radiographic Appearance of the Fore Digit and Carpal Joint in the Mule Foal from Birth to 3 Months of Age.

Few studies have established the normal radiographic anatomical development of the donkey foal and, to date, no data are available for mules. Our aim was thus to evaluate the radiographic development of the fore digit and carpal joint in the mule foal from 0 to 3 months of age. Ten forelimbs of five healthy full-term mule foals were included. Radiographs of the fore digit lateromedial and dorsopalmar and the carpus dorsopalmar were performed weekly for the first month of age, and bi-monthly during the following two months. Fore digit growth plate closure times, morphological, angular, and linear radiographic parameters, and also carpal cuboidal bone mineralization were evaluated. Growth plates were graded as open, closing, and closed. Carpal bone appearance was graded as mature, slightly immature, or immature. Growth plate closure times showed the following: middle phalangeal distal physis (DP) closed at birth, and proximal physis (PP) started to close at around two months; first phalanx DP closed at seven days, and PP started to close at three months; and third metacarpal bone DP started to close at two months. Carpal bones were immature at birth. Distal phalanx (PD) was triangular at birth, palmar processes had developed by one month, and proximal sesamoid bones were trapezoidal by three months. The hoof wall, PD dorsal wall, and palmar angles values changed in parallel, and hoof capsule thickness increased. No PD remodeling evidence within the hoof capsule or variations in the hoof axis with growth were found. We identified a baseline for the interpretation of forelimb radiological features in mule foals. The specific features found were intermediate between those found in horses and donkeys.

Detour test performance of cloned minipigs from three different clone populations.

Genetics, the uterine environment, maternal behavior, and rearing conditions can all influence animal behavioral phenotypes. Some studies on cloned pigs have found no differences between the behavioral patterns of cloned and non-cloned animals. Other studies conducted on dogs have reported similarities in the behavior of cloned subjects. This study evaluated the performance of 12 cloned minipigs from three different clone populations (A, B, C) in a detour test around symmetric and asymmetric barriers. We measured the detour time and patterns, in order to investigate the pigs' cognitive abilities.The detour time and the detour entry/exit pattern were recorded. All the animals tended to keep a fixed entry/exit pattern instead of modifying it to accommodate changes in the working set. Significant differences in detour time were found among the populations, with animals belonging to population B being faster than the others, and also within each population.Our study is one of the few to assess the cognitive abilities of cloned minipigs. The results indicate that even animals belonging to the same cloned population may develop different cognitive, hence behavioral characteristics. Whether cloning can be utilized to obtain similar behavioral phenotypes therefore remains a matter of debate.

Feline Parvovirus Lethal Outbreak in a Group of Adult Cohabiting Domestic Cats.

Feline panleukopenia is a highly contagious and often fatal disease in cats. The virus, known as feline panleukopenia virus (FPV), primarily affects kittens and unvaccinated cats. It is transmitted through contact with infected cats or their bodily fluids, as well as contaminated objects and environments. The diagnosis of FPV infection can be confirmed through a combination of clinical signs, blood tests, and fecal testing. Prevention through vaccination is recommended for all cats. This case report describes an outbreak of feline panleukopenia in a group of unvaccinated domestic cats that resulted in acute mortality. The lesions were evaluated using histopathology, and the specific viral strain was characterized using molecular techniques. The clinical course of the outbreak was peracute, with a hemorrhagic pattern and 100% of lethality. The observed clinical-pathological pattern was unusual; nevertheless, molecular studies did not highlight peculiar genomic features of the parvovirus isolate. The outbreak affected 3 out of 12 cats in a very short time. However, the prompt application of biosecurity measures and vaccination resulted in an effective interruption of virus spread. In conclusion, we could assume that the virus found the ideal conditions to infect and replicate at high titers, resulting in a particularly aggressive outbreak.

A Comparison of the Efficacy of Two Omeprazole Formulations in the Treatment of Equine Gastric Ulcer Syndrome in Racehorses: A Blinded, Randomized Clinical Trial.

Equine gastric ulcer syndrome (EGUS) is the most common disease of the stomach in horses and treatment is based on the oral administration of omeprazole for at least 28 days. Aim of this study was to compare the efficacy of two formulations of oral omeprazole-powder paste and gastro-enteric resistant granules-in the treatment of naturally occurring gastric ulcers in racehorses. Thirty-two adult racehorses, aged between 2 and 10 years old, with clinical signs of EGUS were included in this blinded, randomized clinical trial. Two gastroscopies were performed to evaluate gastric lesions in the squamous or glandular mucosa before and after 28 days of treatment. After the first gastroscopy, 2/32 horses were excluded because affected by equine squamous gastric disease (ESGD) 1/4. The remaining horses were divided into 4 groups, and were treated with 2 formulations of omeprazole (gastroenteric resistant granules, group 1, or powder paste, group 3) or with 2 placebo formulations (granules, group 2, or paste, group 4). Treatments were performed in the placebo horses affected by ESGD or equine glandular gastric disease after the T28 gastroscopy control. No differences were found between groups at T0. Differences were found between T0 vs. T28 for granular (P = .002) and powder paste (P = .01) formulations. No differences were detected between the two groups receiving omeprazole formulations at T28 (0.34) and between T0 vs. T28 for either of the groups receiving placebo treatments. For all the variables, the size of the effect was greater than 0.5, proving that the treatments had a considerable effect. Gastro-enteric resistant granules and powder paste omeprazole showed similar efficacy in the treatment of ESGD. The glandular mucosa responded poorly to treatment with omeprazole.

Autonomic nervous system balance in parturient mares: Spontaneous vs induced delivery.

Delivery is not easily predictable in horses and the consequences of dystocia can be serious for both the mare and foal. An induction protocol with low doses of oxytocin has been reported as a safe procedure. This study investigates the effect of induced delivery on at-term mares' sympathetic-vagal balance. Fourteen mares were included and divided into two groups, one subjected to spontaneous delivery (SD), and one to induced delivery (ID). In both groups, an ECG was recorded using an elastic belt with integrated smart textile electrodes. The recording started before the delivery (Basal), continued close to delivery (Pre-delivery) and during delivery (Delivery), and ended after parturition (Placental expulsion). From the ECGs, Heart Rate Variability (HRV) parameters relating to time and frequency domains and non-linear analysis were extrapolated. The HRV analysis was performed both within the same group (IntraGA) and between the two groups (InterGA). In the present study, spontaneous and induced delivery did not appear to differ in autonomic nervous system functioning. In IntraGA analysis, both for SD and ID mares, delivery and placental expulsion periods were parasympathetic dominated since vagal-related HRV parameters increased. Moreover, no differences were found in InterGA comparison between SD and ID mares, except for the pre-delivery period of ID mares, during which both branches of the autonomic nervous system were activated. These results are in line with the literature on parasympathetic dominance during parturition and no change in Heart Rate Variability following exogenous oxytocin administration in parturient mares.

Breath Alcohol Test Results in Equine Veterinarians after Performing an Abdominal Ultrasound with Ethanol.

Transcutaneous abdominal ultrasonography using alcohol saturation is used in the diagnostic workup of a variety of conditions in horses. The duration of the examination and the amount of alcohol used in each case may vary depending on several factors. The aim of this study is to describe the breath alcohol test results obtained by veterinarians performing abdominal ultrasound on horses. Six volunteers were enrolled, after written consent, and a Standardbred mare was used for the whole study protocol. Each operator performed a total of 6 ultrasounds by pouring the ethanol solution from a jar or by spray application, for a duration of 10, 30, and 60 min. An infrared breath alcohol analyzer was used immediately after completing the ultrasonography and at 5-min intervals until a negative result was obtained. Positive results were obtained for 0-60 min after the procedure. A statistically significant difference was found between the groups that used more than 1000 mL, 300-1000 mL, and less than 300 mL of ethanol. No significant differences were observed between the type of ethanol administration and the time of exposure. Based on this study, equine vets who perform ultrasound on horses can test positive at the breath alcohol test for up to 60 min following ethanol exposure.

A One-Year Retrospective Analysis of Viral and Parasitological Agents in Wildlife Animals Admitted to a First Aid Hospital.

This study aimed to provide information on the presence and frequency of viral and parasitic agents in wildlife presented to a Veterinary Teaching Hospital in 2020-2021. Serum and faecal samples were collected from 50 rescued animals (roe deer, fallow deer, foxes, badgers, pine martens, and porcupines) and examined by serological, molecular, and parasitological techniques. Transtracheal wash (TTW) was also collected post-mortem from roe deer. Overall, the results of the different techniques showed infections with the following viral and parasitic agents: Bovine Viral Diarrhea Virus, Small Ruminant Lentiviruses, Kobuvirus, Astrovirus, Canine Adenovirus 1, Bopivirus, gastrointestinal strongyles, Capillaria, Ancylostomatidae, Toxocara canis, Trichuris vulpis, Hymenolepis, Strongyloides, Eimeria, Isospora, Dictyocaulus, Angiostrongylus vasorum, Crenosoma, Dirofilaria immitis, Neospora caninum, Giardia duodenalis, and Cryptosporidium. Sequencing (Tpi locus) identified G. duodenalis sub-assemblages AI and BIV in one roe deer and one porcupine, respectively. Adult lungworms collected from the TTW were identified as Dictyocaulus capreolus (COX1 gene). This is the first molecular identification of G. duodenalis sub-assemblage AI and D. capreolus in roe deer in Italy. These results show a wide presence of pathogens in wild populations and provide an overview of environmental health surveillance.

Case - Control study: Evaluation of plasma procalcitonin concentration as an indicator of inflammation in healthy and sick cows.

This case - control study aims to evaluate Procalcitonin (PCT) plasma concentrations in healthy and hospitalized cows with a conclusive diagnosis of inflammation due to bacterial infection. Thirty-four healthy and 131 sick cows were included. Procalcitonin concentrations were assessed using an ELISA kit for cattle. Depending on whether sick cows received antimicrobial treatments prior to admission or not, they were divided in treated (TP) or not treated (NTP) subgroups. Mann-Whitney U tests were performed to determine differences between healthy vs sick cows, while Kruskal-Wallis with Dunn's multiple comparison test were applied for healthy vs sick subgroups. Receiver operating characteristic (ROC) analysis was performed to assess the optimal cut-off value. Kaplan-Meier survival curves were determined for cows belonging to the groups with PCT values below and above ROC cut-offs. Plasma PCT concentration was 200.1 (147.8-324.1) pg/mL and 361.6 (239.7-947.1) pg/mL in the healthy control and in the sick group, respectively (P < 0.001). The optimal cut-off value of plasma PCT concentration was 244.4 pg/mL (sensitivity 73.6%, specificity 60.0%). The plasma PCT concentration was 267.5 (210.3-771.2) pg/mL in the TP subgroup and 425.6 (253.1-1242) pg/mL in the NTP subgroup (P = 0.03). Cows with PCT above the ROC cut-off value had a reduced survival percentage and a higher mortality risk (P < 0.05). Procalcitonin showed the ability of differentiate healthy cows from hospitalized cows with a conclusive diagnosis of inflammation due to bacterial infection. Moreover, PCT was a good predictor of negative prognostic outcome.

Respiratory sinus arrhythmia magnitude quantification as a potential marker of stress and pain in cows and sheep.

Respiratory sinus arrhythmia (RSA) provides a quantitative evaluation of 'vagal tone' and may be used for pain and stress assessment in livestock. The aim was to quantify the magnitude of RSA in cows and sheep. In 7 cows and 4 sheep standing at rest we measured the 3-lead electrocardiogram (ECG) together with the pneumogram, to identify inspiration and expiration. For each breath, RSA was the difference in instantaneous heart rate (HR) between the inspiratory peak and the expiratory trough, in percent of mean HR. The resting breathing rates (28 ± 2 and 32 ± 5 breaths/min in cows and sheep, respectively) were about twice those expected for similar size non-ruminants, in conformity with previous reports. Both species had long-period (>15 s) HR fluctuations. The average values of RSA, 1.4 ± 0.2% in cows and 7.8 ± 3.1 in sheep, were lower than those previously computed by an identical approach in humans (12%), dogs (40%) and horses (9%). In conclusion, by breath-by-breath analysis of instantaneous HR we measured RSA in both cows and sheep. Results from the present study represent a preliminary step in assessing whether or not RSA could be used as a biomarker for stress or pain in ruminants.