Efficacy of oral, topical and extended-release injectable formulations of moxidectin combined with doxycycline in Dirofilaria immitis naturally infected dogs.

BACKGROUND: Several studies in both experimentally and naturally infected dogs have reported the adulticide effect of a combination of macrocyclic lactones and doxycycline against Dirofilaria immitis, showing that these protocols can be used as an alternative to melarsomine. The present study evaluated the efficacy of oral, topical and extended-release injectable formulations of moxidectin when combined with doxycycline in dogs naturally infected with D. immitis from a shelter located in southern Italy. METHODS: Thirty dogs with naturally acquired D. immitis infection were divided in three groups (G) and treated with oral moxidectin (G1) once a month for 9 consecutive months, topical moxidectin (G2) once a month for 9 consecutive months or extended release moxidectin injectable (G3) at enrolment and again at 6 months (Day 180). All treatment groups received doxycycline for the first 30 days. Microfilarial concentrations in 1 ml (mff/ml) blood were determined monthly for 9 months with the modified Knott's test. A clinical scoring system was employed for each dog enrolled in the study based on thoracic radiography and cardiac ultrasound (CU) examinations performed at Day - 15 (before treatment) and at Day 180. RESULTS: Results from the present study suggest that the majority of dogs from all treatment groups became antigen negative, as evaluated at Day 270: 9/10 dogs (90.0%) from G1, 6/10 dogs (60.0%) from G2 and 8/10 dogs (80.0%) from G3. Improvement of radiographic alterations was observed in all treatment groups, and almost all dogs were cleared of pulmonary abnormalities by 6 months from the beginning of treatment (P = 0.000). Cardiac ultrasound examination showed a progressive improvement of cardiac function in a limited number of animals (4/30). CONCLUSIONS: The combination of doxycycline and three different formulations of moxidectin leads to antigen-negative status in naturally infected dogs.

Exosomes in Dogs and Cats: An Innovative Approach to Neoplastic and Non-Neoplastic Diseases.

Exosomes are extracellular vesicles with a diameter between 40 and 120 nm, which are derived from all types of cells and released into all biological fluids, such as blood plasma, serum, urine, breast milk, colostrum, and more. They contain proteins, nucleic acids (mRNA, miRNA, other non-coding RNA, and DNA), and lipids. Exosomes represent a potentially accurate footprint of the miRNA profile of the parental cell and can therefore be proposed as potential and sensitive biomarkers, both in diagnosing and monitoring a variety of diseases in humans and animals. Liquid biopsy offers itself as a non-invasive or minimally invasive, pain-free, time-saving alternative to conventional tissue biopsy. Exosomes in both human and veterinary medicine find their major application in neoplastic diseases, but applications in the field of veterinary cardiology, nephrology, reproduction, parasitology, and regenerative medicine are currently being explored. Exosomes can therefore be used as diagnostic, prognostic, and, in some cases, therapeutic tools for several conditions. The aim of this review was to assess the current applications of exosomes in veterinary medicine, particularly in dog and cat patients.

Fast Focus Ultrasound Liver Technique for the Assessment of Cystic Echinococcosis in Sheep.

A complete ultrasound examination (cUS) of the liver was performed on 172 female sheep and compared to the performance of a fast-focused ultrasound technique to diagnose echinococcal cysts. The scanned area was divided in: HYP (right hypocondrium), zone (Z)1 from HYP to the 11th intercostal space (IS), Z2 (10th-8th IS) and Z3 (7th-5th IS). Contiguous scans were also examined (HYP + Z1, Z1 + Z2, Z2 + Z3). Furthermore, during the procedures, the sheep were divided into three groups according to the body weight: Group (G) 1 (lighter), G2 (medium), and G3 (heavier). Finally, diagnostic outcomes were compared with necropsy findings. cUS obtained the highest values of sensitivity (Se) (91%), Specificity (Sp) (80%), and positive-zones (124/138, 90%), as compared to the other scans. cUS was also characterized by high values of Se and Sp and was able to identify a great number of positive-zones, when sheep were divided by body-weight groups. Similar performances were obtained in G1 by HYP (Se: 91%-Sp: 82%; 18/20, 90% of positive-zones) and HYP + Z1 scans (Se: 91%-Sp: 82; 90% of positive-zones, 18/20). Thus, in lighter breeds, the examination of HYP and HYP + Z1 scan windows could be considered reliable techniques for identifying the infected animals, while in heavier sheep the cUS still represents the best option.

Evaluation of the cutaneous inflammatory cells in dogs with leishmaniosis and in dogs without the disease that were naturally infected by Leishmania infantum (syn. L. chagasi).

BACKGROUND: Canine leishmaniosis (CanL) is associated with an aberrant cutaneous immune response. Few studies have compared cutaneous immune cells among dogs with leishmaniosis or infected without leishmaniosis, and noninfected dogs. HYPOTHESIS/OBJECTIVE: Evaluate the number of neutrophils, histiocytes, T lymphocytes, T-bet-positive, GATA3-positive, FoxP3-positive and interleukin (IL)-17-positive cells, in lesional (Group A) and normal-looking (Group B) skin of nine dogs with stage II/III leishmaniosis; in normal-looking PCR-positive (Group C; n = 6) or PCR-negative (Group D; n = 6) skin of infected dogs; and in the normal-looking skin of 12 noninfected dogs (Group E). METHODS AND MATERIALS: Diagnosis of CanL considered clinical signs, clinicopathological abnormalities, detection of Leishmania amastigotes in lymph nodes, and/or bone marrow and positive serological results. Paraffin-embedded skin biopsies were processed for routine immunofluorescence and positive cells were identified using commercially available anti-canine antibodies. RESULTS: In Group A, there was a significantly higher number of neutrophils (P < 0.001), histiocytes (P = 0.012), T lymphocytes (P = 0.011), GATA3-positive (P = 0.02) and IL-17A-positive (P = 0.002) cells compared to Group E. In Group B, there was a significantly higher number of histiocytes (P = 0.02), T lymphocytes (P = 0.004), GATA3-positive (P = 0.006) and FoxP3-positive (P = 0.028) cells compared to Group E. There was no difference in between groups A and B and between groups C or D and E. CONCLUSIONS AND CLINICAL IMPORTANCE: In the lesional and/or normal-looking skin of dogs with moderate/severe CanL there is an infiltration of neutrophils, histiocytes, T lymphocytes, GATA3-, FoxP3- and IL17A-positive cells. By contrast, the number of these cells is not increased in the normal-looking skin of infected dogs without CanL compared to the skin of noninfected dogs.

Identification of differentially expressed microRNAs in the skin of experimentally sensitized naturally affected atopic beagles by next-generation sequencing.

Canine atopic dermatitis (AD) is a very common inflammatory skin disease, but limited data are available on the genetic characterization (somatic mutations, microarrays, and genome-wide association study (GWAS)) of skin lesions in affected dogs. microRNAs are good biomarkers in inflammatory and neoplastic diseases in people. The aim of this study was to evaluate microRNA expression in the skin of atopic beagles, before and after exposure to Dermatophagoides farinae. Four atopic and four unrelated age-matched healthy beagle dogs were enrolled. Total RNA was extracted from flash-frozen skin biopsies of healthy and atopic dogs. For the atopic dogs, skin biopsies were taken from non-lesional (day 0) and lesional skin (day 28 of weekly environmental challenge with Dermatophagoides farinae). Small RNA libraries were constructed and sequenced. The microRNA sequences were aligned to CanFam3.1 genome. Differential expressed microRNAs were selected on the basis of fold-change and statistical significance (fold-change ≥ 1.5 and p ≤ 0.05 as thresholds. A total of 277 microRNAs were sequenced. One hundred and twenty-one differentially regulated microRNAs were identified between non-lesional and healthy skin. Among these, two were increased amount and 119 were decreased amount. A total of 45 differentially regulated microRNAs between lesional and healthy skin were identified, 44 were decreased amount and one was increased amount. Finally, only two increased amount microRNAs were present in lesional skin when compared with that of non-lesional skin. This is the first study in which dysregulation of microRNAs has been associated with lesional and non-lesional canine AD. Larger studies are needed to understand the role of microRNA in canine AD.

ACVIM consensus statement on the diagnosis of immune-mediated hemolytic anemia in dogs and cats.

Immune-mediated hemolytic anemia (IMHA) is an important cause of morbidity and mortality in dogs. IMHA also occurs in cats, although less commonly. IMHA is considered secondary when it can be attributed to an underlying disease, and as primary (idiopathic) if no cause is found. Eliminating diseases that cause IMHA may attenuate or stop immune-mediated erythrocyte destruction, and adverse consequences of long-term immunosuppressive treatment can be avoided. Infections, cancer, drugs, vaccines, and inflammatory processes may be underlying causes of IMHA. Evidence for these comorbidities has not been systematically evaluated, rendering evidence-based decisions difficult. We identified and extracted data from studies published in the veterinary literature and developed a novel tool for evaluation of evidence quality, using it to assess study design, diagnostic criteria for IMHA, comorbidities, and causality. Succinct evidence summary statements were written, along with screening recommendations. Statements were refined by conducting 3 iterations of Delphi review with panel and task force members. Commentary was solicited from several professional bodies to maximize clinical applicability before the recommendations were submitted. The resulting document is intended to provide clinical guidelines for diagnosis of, and underlying disease screening for, IMHA in dogs and cats. These should be implemented with consideration of animal, owner, and geographical factors.

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.

Monitoring multiple myeloma by idiotype-specific peptide binders of tumor-derived exosomes.

Tumor-derived exosomes (TDEs) play a pivotal role in tumor establishment and progression, and are emerging biomarkers for tumor diagnosis in personalized medicine. To date, there is a lack of efficient technology platforms for exosome isolation and characterization. Multiple myeloma (MM) is an incurable B-cell malignancy due to the rapid development of drug-resistance. MM-released exosomes express the immunoglobulin B-cell receptor (Ig-BCR) of the tumor B-cells, which can be targeted by Idiotype-binding peptides (Id-peptides). In this study, we analyzed the production of MM-released exosomes in the murine 5T33MM multiple myeloma model as biomarkers of tumor growth. To this end, we selected Id-peptides by screening a phage display library using as bait the Ig-BCR expressed by 5T33MM cells. By FACS, the FITC-conjugated Id-peptides detected the MM-released exosomes in the serum of 5T33MM-engrafted mice, levels of which are correlated with tumor progression at an earlier time point compared to serum paraprotein. These results indicate that Id-peptide-based recognition of MM-released exosomes may represent a very sensitive diagnostic approach for clinical evaluation of disease progression.

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.

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 manufacturer's 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.

Situs inversus totalis associated with subaortic stenosis, restrictive ventricular septal defect, and tricuspid dysplasia in an adult dog.

A rare association between situs inversus totalis (SIT), restrictive ventricular septal defect, severe subaortic stenosis, and tricuspid dysplasia was observed in an adult mixed-breed dog. Primary ciliary dyskinesia and Kartagener's syndrome were excluded. After 15 mo the dog died suddenly. The association between SIT and congenital heart diseases is discussed.

The effects of prednisone on haemostasis in leishmaniotic dogs treated with meglumine antimoniate and allopurinol.

Thirty dogs naturally infected with Leishmania infantum were studied in order to determine the effects of treatment on haemostatic function. The animals were divided randomly into two treatment groups: Group 1 received meglumine antimoniate and allopurinol; Group 2 dogs were given the same treatment plus prednisone. Ten healthy animals were used as untreated controls. Clinical examination and determination of platelet aggregation, coagulation factors and biochemical parameters were undertaken before treatment and after 15, 30 and 60 days. A significant improvement in platelet aggregation was detected after 60 days in Group 1, but only after 15 days in Group 2. In both treated groups, platelet aggregation was lower than in the control group at the end of the study. The results suggest that prednisone may be a useful tool in the treatment of haemostatic disorders during canine leishmaniosis. The potential benefits and risks due to the use of corticosteroids in the treatment of leishmaniosis are discussed.