Jacopo Guccione

Seroprevalence and risk factors of Neospora spp. in donkeys from Southern Italy

In some European countries there is an increasing interest on donkey. Despite there are few data regarding the donkeys parasitic diseases especially those with a protozoal etiology as neosporosis. Samples used in the study were collected from 238 domestic donkeys during year 2010 in Southern Italy from 207 females and 31 males of five breeds (Martina-Franca, Amiata, Sicilian-Grey, Ragusano, Sardinian) and crossbreeds with the average age 9 years (1 month - 24 year). Sera were tested by a competitive-inhibition enzyme-linked immunosorbent assay for antibodies against Neospora caninum; the sera were marked positive, if more than 30% inhibition was found. Out of a total 238 donkeys, 28 (11.8%) were found positive for Neospora antibodies with 12% in females and 6% in males. Different seroprevalence 15.4%, 16%, 12% and 8.8% were found in age categories <1 year, 1-4 years, 5-9 years and ≥10 years, respectively. The seroprevalence ranged in different breeds from 36% (Sicilian-Grey) to 0% (Sardinian) and in different use from 17% (for breeding) to 0% (for meat production). Logistic regression analysis demonstrated evidence of a significant (P<0.05) association between crossbreed origin of samples and risk of protozoan infection; age of donkeys was also significant risk factor for protozoan infection. No statistical significant difference (P>0.05) was found among genders and use of donkeys and risk of N. caninum infection. This is the first serological survey for Neospora spp. performed in donkeys.

Short communication: Effects of systemic treatment with penethamate hydriodide on udder health and milk yields in dry primiparous Mediterranean buffaloes (Bubalus bubalis)

The effects of penethamate hydriodide (Mamyzin, Boehringer Ingelheim, Ingelheim, Germany) on udder health and milk yields were evaluated in primiparous Mediterranean buffaloes (Bubalus bubalis). An intramuscular administration of 10 million international units was performed in 20 buffaloes at 7 d precalving (treatment group; TG), and 20 animals were enrolled as the control group (CG). Evening milk samplings were performed at 10, 30, and 60 d in milk (DIM). Somatic cell count (SCC) values were evaluated on composite milk samples, whereas bacteriological culture and California Mastitis Test were performed on quarter milk. Daily milk yields were recorded after all milkings. After 60 DIM, composite milk samples from each animal were collected for monthly SCC and bacteriological culture until drying off. Statistically significant differences were found between the prevalence of mastitic quarters in the 2 groups at 10 and 30 DIM, and between the incidence of mastitic animals during the examined period (TG: 4/20, 20% vs. CG: 10/20, 50%). Even though lower and higher values of SCC and milk yields were found in TG during each sampling, statistically significant differences were only found at 30 (SCC) and 60 DIM (milk yields). In our study, the antibiotic administration precalving showed good bactericidal activity against the most common udder-specific pathogens that cause mastitis in primiparous Mediterranean buffaloes, and greater efficacy was observed at 10 and 30 DIM compared with 60 DIM. Given the significant decrease in SCC and increase in yields achieved, use of this antibiotic could be economically beneficial in buffalo breeding.

Serum biochemistry profile, inflammatory cytokines, adipokines and cardiovascular findings in obese dogs.

The aim of this study was to evaluate the serum biochemistry profile, inflammatory cytokines, adipokines and cardiovascular findings in obese dogs. Twenty obese and 20 normal weight healthy pet dogs were recruited into the study, where they underwent blood testing and assessment of cardiovascular function (blood pressure analysis, electrocardiography and echocardiography). Higher concentrations of total cholesterol, triglycerides, lactate dehydrogenase, total serum proteins, α-globulins, total bilirubin, insulin, insulin:glucose ratio, alkaline phosphate and alanine aminotransferase were observed in obese dogs than dogs of normal weight. There were no differences in concentrations of tumour necrosis factor (TNF)-α or interleukin (IL)-6 between the two groups. Obese dogs had higher serum leptin but lower adiponectin concentrations than dogs of normal weight. Systolic arterial blood pressure was higher in obese dogs than dogs of normal weight. The values for the thickness of the free wall of the left ventricle and interventricular septal thickness were greater at end-diastole in obese dogs compared to dogs of normal weight. Four of 20 obese dogs were determined to have obesity-related metabolic dysfunction (ORMD). The findings indicate that a chronic inflammatory state is not necessarily evident in obese dogs, as has been described in human beings, and the criteria used for ORMD can be used to define this syndrome in dogs. In this study, canine obesity was associated with cardiac and vascular dysfunction.

Clotting profile in cattle showing chronic enzootic haematuria (CEH) and bladder neoplasms

Primary haemostasis (bleeding and blood clotting time), activated partial thromboplastin time (APTT), prothrombin time (PT), antithrombin III (ATIII), protein C, protein S, fibrinogen and D-dimer were determined in 13 cattle affected by chronic enzootic haematuria (CEH) and bladder neoplasms and 10 healthy cattle (control group). Increases in antithrombin III and protein S activities (P<0.01) and protein C and fibrinogen plasma levels (P<0.05) were observed in sick animals, while activated partial thromboplastin time, prothrombin time, and D-dimer did not show significant differences when compared to healthy animals. The clotting profile observed does not seem responsible for the chronic bleeding typical of CEH. The observed modification of some coagulation markers may derive from multiple interactions among cancer, inflammation and viral infection status typical of this syndrome.

Field efficacy of eprinomectin against the sucking louse Haematopinus asini on naturally infested donkeys.

A trial was conducted to assess the efficacy of eprinomectin (EPR) against the sucking louse Haematopinus asini on naturally infested donkeys. Parasitological investigations were performed on fifteen animals. On day 0, donkeys received EPR pour-on at the manufacturers recommended cattle dose. Louse counts were performed on days -1, 7, 14, 21, 28, 35, 42, 49 and 56 at seven predilection sites on the skin of each donkey. EPR was completely effective (100%) from day 7, until the end of the study. Clinically no adverse reactions were observed in any of donkeys treated. EPR was considered to be 100% effective against H. asini. This is the first trial to evaluate the efficacy of EPR against a natural louse infestation in donkeys.

Increased leptin mRNA expression in the blood of dogs naturally infected by Leishmania infantum.

Canine leishmaniosis (CL) is a severe and potentially fatal zoonosis caused by the protozoan Leishmania infantum. Severe forms of CL are commonly associated with a non-protective, humoral immune-response and high parasitic loads. Leptin, a 16 kD hormone mainly secreted by adipocytes, regulates both the innate and adaptive immunity. The goal of this study was to evaluate leptin mRNA expression levels in blood samples from privately owned dogs with CL (n = 11) and healthy controls (n = 10) using quantitative, real-time polymerase chain reaction. Blood samples from dogs with CL expressed significantly higher leptin mRNA levels (two-fold) compared to healthy controls (P = 0.018). The results suggest a possible involvement of leptin in the pathophysiology of Leishmania infection in dogs and the possible use of leptin as a biomarker for CL. Future studies investigating the immunological role of leptin in dogs with CL are warranted.

Validation of a pedometer algorithm as a tool for evaluation of locomotor behaviour in dairy Mediterranean buffalo

This research communication validates an algorithm to monitor natural occurrence of locomotor behaviours in dairy Mediterranean buffalo based on the output of a 3-dimensional accelerometer (RumiWatch®, pedometer). Several characteristics of the locomotor behaviour were detected with a very high (up-right, lying and standing time) or high degree of correlation (walking time and number of strides) and a low mean difference with the video recording. The proportion of correctly detected events exceeded 99 % for the following variables: stand up and lie down events, as well as number of lying, standing or walking bouts. The mean relative measurement error was less than 10 % for the variables: lying, standing, up-right times and number of strides as compared with gold standard. This new algorithm may represent the base for a future early and real-time disease warning system aiming to gain higher health standard in these ruminants.

Circulating regulatory T cells (Treg), leptin and induction of proinflammatory activity in obese Labrador Retriever dogs

Over-nutrition and obesity have been associated with impaired immunity and low-grade inflammation in humans and mouse models. In this context, a causal role for unbalanced T regulatory cell (Treg)-dependent mechanisms has been largely suggested. Obesity is the most common nutritional disorder in dogs. However, it is not defined whether canine obesity may influence circulating Treg as well as if their number variation might be associated with the occurrence of systemic inflammation. The present study investigated the immune profile of healthy adult obese dogs belonging to the Labrador Retriever breed, in comparison with the normal weight counterpart. Indeed, obesity has been described as particularly evident in this dogs. With this purpose, 26 healthy dogs were enrolled and divided into two groups based on body condition score (BCS): controls (CTR: BCS 4-5) and obeses (OB: BCS ≥ 7). Our data indicate that adult obese Labrador Retrievers are characterised by the inverse correlation between leptin serum concentration and circulating Treg (CD4+CD25highFoxp3+) levels. In addition, an increased number of cytotoxic T cell effectors (CD3+CD8+) and a higher IFN-γ production by cytotoxic T lymphocytes were observed in OB group. These results may provide new insights into the immunological dysregulation frequently associated to obesity in humans and still undefined in dogs.

Clinical efficacy and pharmacokinetics of meloxicam in Mediterranean buffalo calves (Bubalus bubalis)

The aims of the investigation were to establish for the first time (i) clinical efficacy and (ii) pharmacokinetic profile of meloxicam intravenously (IV) administered in male Mediterranean buffalo calves after surgical orchiectomy. The study was performed on 10 healthy buffalo calves, between 4 and 5 months old and between 127 and 135 kg of body weight (b.w.). An IV injection of 0.5 mg/kg b.w. of meloxicam was administered in six calves (treated group, TG) immediately after surgery; the other four animals were used as untreated control group (CG). The clinical efficacy of meloxicam was evaluated pre- and post-surgery by monitoring respiratory rate (RR), heart rate (HR), rectal temperature (T°C), serum cortisol levels (SCL) and pain score (PS). Significant inter-groups differences were detected at sampling times (T): 4 hour (h) for RR (P<0.05), at T1-4-6-8 h for PS (P<0.05) and at T4-6-8 h for SCL (P < 0.0001). Regarding the mean intra-group values observed pre (T0) and post-surgery (from T15 min to T72 h), significant difference between the groups were found for RR (P<0.01), PS and SCL (P<0.05). The pharmacokinetic profile was best fitted by a two-compartmental model and characterized by a fast distribution half-life and slow elimination half-life (0.09 ± 0.06 h and 21.51 ± 6.4 h, respectively) and meloxicam mean concentrations at 96 h was of 0.18 ± 0.14 μg/mL. The volume of distribution and clearance values were quite low, but reasonably homogenous among individuals (Vdss 142.31 ± 55.08 mL/kg and ClB 4.38 ± 0.95 mL/kg/h, respectively). The IV administration of meloxicam in buffalo calves shows encouraging effects represented by significant and prolonged analgesic effects, significant reduction of SCL as well as similar pharmacokinetic profile to bovine calves.

Pharmacokinetics and clinical efficacy of meloxicam in Italian Mediterranean Buffalo calf (Bubalus bubalis)

Introduction: The anatomy and physiology of pigs is closely related to human characteristics. Therefore, the use of pigs as an animal model to study the pharmacokinetic (PK) behavior of drugs in therapeutic subpopulations, including pediatrics, could be of interest. One of the key PK processes, biotransformation, is primarily mediated by the cytochrome P450 (CYP) enzyme system. Literature reports have demonstrated a high homology between human and porcine CYP3A, 2C and 2E in adults, namely at least 75, 62 and 79% amino acid sequence identity, respectively. However, data regarding the ontogeny of porcine CYP enzymes are lacking. Therefore, in order to assess whether piglets might serve as a model for pediatric PK studies, knowledge regarding the ontogeny of the CYP enzymes in pigs is mandatory. Materials and methods: Liver samples were collected immediately after euthanasia from 16 pigs (8 males and 8 females, Hybrid sow x Pietrain boar) of different ages (2 days, 4 and 8 weeks, 6 months-old). Samples were snap-frozen and stored at <-80°C until analysis. Microsomes were prepared by a differential centrifugation method. Midazolam, tolbutamide and chlorzoxazone probe drugs were used to determine the in vitro CYP3A, 2C and 2E catalytic activity, respectively. The corresponding metabolites, namely 1-hydroxy-midazolam, 4-hydroxy-tolbutamide and 6-hydroxy-chlorzoxazone, were quantified using a validated UHPLC-MS/MS method (1). Furthermore, the microsomal protein per gram of liver was determined as it is an important scaling factor in the extrapolation of the obtained in vitro enzyme activities to in vivo (2). Results and conclusions: The biotransformation of midazolam, tolbutamide and chlorzoxazone increased with age. The mean (±SD) CYP3A activity was 60.5 (±45.7) and 83.3 (±20.7) pmol/min/mg protein at the age of 2 days, 971.1 (±367.8) and 1072.7 (±371.7) pmol/min/mg protein at the age of 4 weeks and 723.4 (±146.3) and 1134.7 (±282.6) pmol/min/mg protein at 8 weeks of age for the barrows and sows, respectively. CYP2C activity at the same ages increased from 20.1 (±12.3) and 29.1 (±18.5) to 78.3 (±25.6) and 106.7 (±69.1) and 103.5 (±39.6) and 170.2 (±71.9) pmol/min/mg protein, while the activity of CYP2E was 539.3 (±251.0) and 643.3 (±220.3), 747.7 (±134.8) and 948.9 (±246.2) and 957.9 (±221.7) and 1549.8 (±345.0) pmol/min/mg protein, respectively for the barrows and sows. Significant sex differences (P<0.05) were only observed at 8 weeks of age. These data show similar trends with human CYP ontogeny.