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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. immitisfrom a shelter located in southern Italy. Methods A total of 30 dogs with naturally acquired D. immitis infection were divided in three groups (G) and treated either with oral moxidectin (G1) once a month for 9 consecutive months, topical moxidectin (G2) once a month for 9 consecutive months or with an 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 concentration in 1 ml (mff/ml) of 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) exams 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 six 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.

Background Next-Generation-Sequencing (NGS) enables detection of microorganisms present in biological and other matrices of various origin and nature, allowing not only the identification of known phyla and strains but also the discovery of novel ones. The large amount of metagenomic shotgun data produced by NGS require comprehensive and user-friendly pipelines for data analysis, that speed up the bioinformatics steps, relieving the users from the need to manually perform complex and time-consuming tasks. Results We describe here HOME-BIO (sHOtgun MEtagenomic analysis of BIOlogical entities), an exhaustive pipeline for metagenomics data analysis, comprising three independent analytical modules designed for an inclusive analysis of large NGS datasets. Conclusions HOME-BIO is a powerful and easy-to-use tool that can be run also by users with limited computational expertise. It allows in-depth analyses by removing low-complexity/ problematic reads, integrating the analytical steps that lead to a comprehensive taxonomy profile of each sample by querying different source databases, and it is customizable according to specific users’ needs.

The research was focused on the surveillance of the exposure of buffalo milkers in contact with both animals and potentially contaminated equipment, pointing attention on the diffusion of antibiotic-resistant

The research was focused on the surveillance of the exposure of buffalo milkers in contact with both animals and potentially contaminated equipment, pointing attention on the diffusion of antibiotic-resistant Staphylococcus spp. The monitoring was performed for 12 months, allowing the collection of 600 raw milk and buffalo udder surface samples, 192 milking lanes, 400 milking clusters, 160 personal protective equipment (PPEs) and electronic devices surface samples in contact with the workers of four milking parlors located in Southern Italy. The analysis of the milk samples evidenced the highest exposure to the bacteria considered (and mainly to S. aureus) from late winter–spring seasons onward. The possible risk arising from buffalo udder, milking clusters, and lines were instead considered rather stable along the entire period of sampling. The PPEs turned out to be a source of contamination for milkers mainly during the spring and summer periods. The analysis for oxacillin/methicillin resistance revealed in all the farms enrolled an overall amount of 37.5% of Staphylococci strains (belonging to S. aureus, S. haemolyticus, S. pseudintermedius, S. chromogenes species) resistant both to methicillin and oxacillin. The investigation demonstrated that the potential transfer of pathogenic bacteria to humans would have a better chance to occur at milk resumption time (since late winter–spring onward) when the number of animals to be milked is greater and the activity in the milking parlor is more challenging. At the same time, the findings seem to point out that the potential risk may be worsened by a significant presence of oxacillin/methicillin-resistant Staphylococci, potentially resulting from irrational use of antibiotics.

The study evaluated the dynamics of macrophages and polymorphonuclear leukocytes milk-secreted by Mediterranean Buffaloes (MBs). Sixty quarter-milk-samples were collected and divided into three groups (n = 20 units each one): clinical mastitis (CM), subclinical mastitis (SCM), and intramammary infection (IMI). The control group consisted of an additional 20 healthy quarters. Their health status was assessed by clinical examination, quantitative somatic cell count (QSCC) and bacteriological milk culture. Finally, a differential somatic cell count (DSCC) was performed on all the milk samples. The mean percentage of macrophages, both in CM- and SCM-quarters, showed a significant difference as compared with the healthy-ones. Significant differences were also detected comparing the mean percentages of polymorphonuclear leukocytes between CM- and healthy-quarters, SCM and healthy, IMI and healthy. The QSCC revealed a weak-significant-negative-correlation with the quantitation of macrophages (r = −0.388), and a moderate-significant-positive-correlation with the polymorphonuclear leukocytes (r = 0.477). Macrophages and polymorphonuclear leukocytes showed a weak-significant-negative-correlation between them (r = −0.247). The interpretation of macrophages and polymorphonuclear leukocytes dynamics in milk provided beneficial information regarding the clinical status of the quarters enrolled. Future studies exploring the potential use of DSCC to improve udder health represent an interesting perspective in these ruminants.

Objectives To identify the predictive value on time to onset of heart failure (HF) or cardiac death of clinical, radiographic, and echocardiographic variables, as well as cardiac biomarkers N-terminal pro brain natriuretic peptide (NT-proBNP) and cardiac troponin I in dogs with preclinical myxomatous mitral valve disease (MMVD). Animals One hundred sixty-eight dogs with preclinical MMVD and left atrium to aortic root ratio ≥1.6 (LA:Ao) and normalized left ventricular end-diastolic diameter ≥1.7 were included. Methods Prospective, randomized, multicenter, single-blinded, placebo-controlled study. Clinical, radiographic, echocardiographic variables and plasma cardiac biomarkers concentrations were compared at different time points. Using receiving operating curves analysis, best cutoff for selected variables was identified and the risk to develop the study endpoint at six-month intervals was calculated. Results Left atrial to aortic root ratio >2.1 (hazard ratio [HR] 3.2, 95% confidence interval [95% CI] 1.9–5.6), normalized left ventricular end-diastolic diameter > 1.9 (HR: 6.3; 95% CI: 3.3–11.8), early transmitral peak velocity (E peak) > 1 m/sec (HR: 3.9; 95% CI: 2.3–6.7), and NT-proBNP > 1500 ρmol/L (HR: 5.7; 95% CI: 3.3–9.5) were associated with increased risk of HF or cardiac death. The best fit model to predict the risk to reach the endpoint was represented by the plasma NT-proBNP concentrations adjusted for LA:Ao and E peak. Conclusions Logistic and survival models including echocardiographic variables and NT-proBNP can be used to identify dogs with preclinical MMVD at higher risk to develop HF or cardiac death.

Obesity is an important health issue in dogs since it influences a plethora of associated pathologies, including dermatological disorders. Considering the scarcity of information in pets, this work aimed to evaluate the localization and expression of adiponectin (ADIPOQ) and its two receptors (ADIPOR1 and ADIPOR2) in the skin of 10 obese dogs, compared with serum ADIPOQ level. Through immunohistochemistry, ADIPOQ and ADIPOR2 were observed in the adipose tissue, sweat and sebaceous glands, endothelium, and some connective cells. Both receptors were observed in the epidermis and the hair follicles, other than in the sweat and sebaceous glands. Real-time PCR evidenced that the ADIPOQ and ADIPOR2 transcripts were expressed 5.4-fold (p &lt, 0.01) and 2.3-fold less (p &lt, 0.01), respectively, in obese than in normal weight dogs, while ADIPOR1 expression did not change. Obese dogs showed lower serum ADIPOQ levels than the normal weight group. Accordingly, ADIPOQ and ADIPOR2 expression in the skin appear negatively correlated with obesity in the same way as the serum ADIPOQ level. These findings evidence that ADIPOQ system changes in the skin of obese dogs and suggest that the ADIPOQ effect on the skin is at least in part regulated by the reduced expression of ADIPOR2.

Cystic echinococcosis (CE) is a severe zoonosis, caused by the larval stage of the tapeworm Echinococcus granulosus. This helminth infection is of increasing public health and socio-economic concern due to the considerable morbidity rates that cause economic losses both in the public health sector and in the livestock industry. Control programmes against E. granulosus are considered long-term actions which require an integrated approach and high expenditure of time and financial resources. Since 2010, an integrated approach to control CE has been implemented in a highly endemic area of continental southern Italy (Campania region). Innovative procedures and tools have been developed and exploited during the control programme based on the following strategies: i) active and passive surveillance in livestock (using geospatial tools for georeferencing), ii) diagnosis in dogs (using the FLOTAC techniques and molecular analysis), iii) targeted treatment of farm dogs (using purpose-built confinement cages), iv) early diagnosis in livestock (by ultrasonography), v) surveillance in humans (through hospital discharge records analysis), vi) monitoring the food chain (analysing raw vegetables), vii) outreach activities to the general public (through dissemination material, e.g. brochures, gadgets, videos, virtual reality). Over eight years, the integrated approach and the new strategies developed have resulted in a noteworthy reduction of the parasite infection rates in livestock (e.g. up to 30 % in sheep). The results obtained so far highlight that using a one health multidisciplinary and multi-institution effort is of pivotal importance in preparing CE control programmes at regional level and could be extended to other endemic Mediterranean areas.

Background Mastitis is one of the most costly diseases in Mediterranean buffalo (MB). At present, just a few specific antibiotics registered for this dairy specie have been synthetized. Efficacy of an antibiotic dry buffalo therapy (aDBT) against Staphylococcus aureus (S. aureus) mastitis, based on intra-quarter administration of 600 mg of benzathine cloxacillin, have been evaluated for the first time. Eighty MB’s quarters received a drying-off therapy (aDBT-group) and 80 were left untreated (no-aDBT-group). They were sampled at drying-off (pre-treatment) and at the resumption of milking [Fresh calver mastitis rate, dry period new mastitis rate, dry period cure rate, and persistent mastitis rate were calculated for clinical monitoring. Overall proportion of positive quarters/animals, quarters affected by mastitis or intramammary infections (IMI), effects on somatic cell count (SCC) and milk yield were also assessed. Results An inter-group difference (aDBT vs. no-aDBT) was recorded for all the indexes considered. An intra-group (drying-off vs. S. aureus was observed. Regarding the subclinical ones, a higher intra-group difference was observed in aDBT than no-aDBT group, while an inter-group difference was recorded at second sampling. No protective effect was observed against IMI. SCC showed an inter-group difference at second sampling, while none difference was instead detected for milk yield. Conclusions The effects against S. aureus mastitis of benzathine cloxacillin administration at drying-off were assessed for the first time in MB. Its use shows encouraging results in reducing the proportion of mastitis and positive animals at the resumption of the lactation.

One hundred twenty-four water buffaloes (Bubalus bubalis) originating from 9 farms locatedin southern Italy were tested to investigate simultaneously, for the first time, the seroprevalence ofthe protozoaNeospora caninum, Toxoplasma gondiiandBesnoitia besnoitiby ELISA tests and to evaluatethe clinical findings potentially associated to the presence of these aborting parasitic pathogens.Twenty-five of 124 buffaloes (20.2%) were positive forN. caninum, while 17/124 (13.7%) forT. gondii.No buffalo showed specific antibodies forB. besnoiti.Nineteen of 124 animals (15.3%) were foundseropositive for bothT. gondiiandN. caninum. The univariate statistical analysis showed that theseroprevalence ofN. caninumis significantly associated with abortion and presence of retained foetalmembranes, while the seroprevalence ofT. gondiiis significantly associated with an increase ofdays open. The logistic regression models showed that the co-infection byN. caninumandT. gondiistrengthened the abortive effects (OR=7.330) and showed further negative effects on the parameterembryonic death (OR=2.607). The outcome revealed herein represents a high exposure ofN. caninumandT. gondiiin water buffaloes with reproduction disorders that deserves attention for both economicreasons, animal health and welfare (PDF) Seroprevalence and Clinical Outcomes of Neospora caninum, Toxoplasma gondii and Besnoitia besnoiti Infections in Water Buffaloes (Bubalus bubalis). Available from: https://www.researchgate.net/publication/340129008_Seroprevalence_and_Clinical_Outcomes_of_Neospora_caninum_Toxoplasma_gondii_and_Besnoitia_besnoiti_Infections_in_Water_Buffaloes_Bubalus_bubalis [accessed Mar 24 2020].

Aquaporins (AQPs) are a family of thirteen membrane proteins that play an essential role in the transport of fluids across the cell plasma membrane. Recently, the expression of AQPs in different ocular tissues and their involvement in the pathophysiology of eye diseases, have garnered attention. Considering that literature on AQP expression in the lacrimal glands and their secretion is scarce, we aimed to characterise AQP1 expression in the tears of healthy dogs using two tear collection methods (Schirmer tear strips (STS) and ophthalmic sponges (OS)). Fifteen healthy dogs, free of ophthalmic diseases, were included in the study. Tear collection was performed by using STS in one eye and OS in the other. After the extraction of proteins from the tears, the expression of AQP1 was analysed by Western blotting. AQP1 was expressed as a band of 28 kDa. In addition, differences were observed in the expression of AQP1 and in the correlation between tear volume and protein concentration, in tears collected by the two different methods. Our results suggest that AQP1 has a specific role in tear secretion, further research is required to assess its particular role in the function of the ocular surface in eye physiology and pathology.

Current knowledge on the role of exosomal microRNA (miRNA) in canine leishmaniasis (CL), with particular regards to the interaction between miR-122 and lipid alterations, is limited. The aim of this study was to isolate/characterize exosomes in canine serum and evaluate the expression of miR-122 in ten healthy and ten leishmaniotic dogs. Serum exosomes were isolated using a polymer-based kit, ExoQuick&reg, and characterized by flow cytometry and transmission electron microscopy, whereas miR-122-5p expression was evaluated by quantitative reverse-transcriptase polymerase chain reaction. A significant decreased expression of exosomal miR-122-5p, decreased serum levels of high-density lipoproteins, and increased serum levels of low-density lipoproteins were seen in leishmaniotic dogs when compared with healthy dogs. These results suggest that hepatic dysfunctions induced by the parasite interfere with lipoprotein status. The decreased expression of exosomal miR122 represents an additional effect of Leishmania infection in dogs as in people.

Introduction: Efficacy of renin-angiotensin-aldosterone system (RAAS) blockade using angiotensin-converting enzyme inhibitors (ACEi) in dogs with preclinical myxomatous mitral valve disease (MMVD) is controversial. Hypothesis: Administration of spironolactone (2-4 mg q 24 h) and benazepril (0.25-0.5 mg q 24 h) in dogs with preclinical MMVD, not receiving any other cardiac medications, delays the onset of heart failure (HF) and cardiac-related death. Moreover, it reduces the progression of the disease as indicated by echocardiographic parameters and level of cardiac biomarkers N-terminal pro brain natriuretic peptide (NT-proBNP) and cardiac troponin I (cTnI). Animals: 184 dogs with pre-clinical MMVD and left atrium-to-aortic root ratio (LA:Ao) >= 1.6 and normalized left ventricular end-diastolic diameter (LVEDDn) >= 1.7. Methods: This is a prospective, randomized, multicenter, single-blinded, placebo-controlled study. Primary outcome variable was time-to-onset of first occurrence of HF or cardiac death. Secondary end points included effect of treatment on progression of the disease based on echocardiographic and radiographic parameters, as well as variations of NT-proBNP and cTnI concentrations. Results: The median time to primary end point was 902 days (95% confidence interval (CI) 682-not available) for the treatment group and 1139 days (95% CI 732-NA) for the control group (p = 0.45). Vertebral heart score (p = 0.05), LA:Ao (p < 0.001), LVEDDn (p < 0.001), trans-mitral E peak velocity (p = 0.011), and NT-proBNP (p = 0.037) were lower at the end of study in the treatment group. Conclusions: This study failed in demonstrating that combined administration of spironolactone and benazepril delays onset of HF in dogs with preclinical MMVD. However, such treatment induces beneficial effects on cardiac remodeling and these results could be of clinical relevance. (C) 2020 The Authors. Published by Elsevier B.V.

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.

A recently developed bovine cathelicidin (CATH) ELISA was evaluated in the dairy buffalo (Bubalus bubalis) by testing 618 quarter milk samples from a herd with subclinical mastitis cases. Somatic cell count (SCC) and bacteriological culture (BC) were carried out on the same samples for comparison. Out of 618 quarters, 258 (41.75%) were positive to CATH, 289 (46.76%) had SCC > 200,000 cells/mL, and 457 (73.95%) were positive to BC. The most prevalent microorganism was Staphylococcus aureus (SAU, 35.76% of all quarters), followed by non-aureus staphylococci (NAS, 22.17% of all quarters). Clinical mastitis quarters were only 7 (1.13%). CATH levels were significantly higher in clinical quarters and in high SCC, BC-positive quarters than in healthy, low SCC, BC-negative quarters. The highest median values were observed for SAU and the lowest for NAS. Differences among microorganism classes were generally more significant for SCC than for CATH. Test char acteristics of the CATH ELISA, evaluated by considering as true positives all BC-positive quarters with SCC > 200,000 cells/mL (N = 242), and as true negatives all sterile quarters with SCC < 200,000 cells/mL (N = 44), were as follows: sensitivity 57.85%, specificity 84.09%, positive predictive value 95.24%, negative predictive value 26.62%, accuracy 61.89%. Therefore, the bovine CATH ELISA showed a fair sensitivity and a good specificity in detecting water buffalo mastitis.

This research communication addresses the goal of validating an algorithm to monitor natural occurrence of feeding behaviours in dairy Mediterranean buffalo based on the output of a noseband pressure sensor (RumiWatch®, halter). Several characteristics of the feeding behaviour were detected with a very high (ruminating boluses), high (chews per bolus) and moderate degree of correlation (chews per minute) with video analyses (gold standard). All of them were associated with a low mean difference with the gold standard, and the mean relative measurement error ranged between low (ruminating boluses) and moderate (chews per bolus and chews per minute). The proportion of correctly detected events for the variables rumination and eating time was 98 and 99%, respectively. The collection of data and subsequent evaluation of the parameters investigated may provide objective information on Mediterranean Buffalo behaviours allowing for reliable studies of the animal welfare in this ruminant in the future.

One hundred and twenty dogs were enrolled to value the effect of loading condition changes on left ventricular volumes before and 24-hours after the patent ductus arteriosus (PDA) occlusion by Amplatzer Canine Duct Occluder (ACDO) using standard echocardiography. The animals were divided in pure breed (n. 94) and mixed breed (n. 26); subsequently, the pure breed dogs were divided on the basis of the size of the breed of belonging in 3 groups (small size n. 36; medium size n. 8; large size n. 50). Moreover, the animals were divided in three classes based on their age: until 6 months; 6-12 months; over 12 months. A significant reduction of all the examined parameters (left ventricle internal diameter at end-diastole-LVIDd; left ventricle internal diameter at end-systole-LVIDs; end-diastolic volume-EDV; end-systolic volume-ESV; end-diastolic volume index-EDVI; end-systolic volume index-ESVI; fractional shortening-FS) was observed after ductal closure. Twenty-four hours after the closure, the evaluation of the relative percentage difference (RPD) of the echocardiographic parameters showed a significant reduction, higher in small size breed than in large size breed dogs. No significant difference related to breed size was observed only for RPD_FS variable. A significant interaction effect, between breed size and age classes, was observed only for RPD_EDVI (F = 3.39; p = 0.039). Until six months of age there was no significant difference in RPD_EDVI reduction, but over 6 months a significant reduction between small size and large size breed dogs at 24-hours from the occlusion was observed. In conclusion, our data seem to indicate that small breed dogs show a greater tolerance to congenital volume overload than large breed dogs, and this finding could be justify a delay of PDA closure in order to simplify the interventional procedure.

Simple Summary Obesity is a widespread phenomenon in pets and its growing trend is similar to the human one. It can be associated with skin pathologies but there is little information on this field in domestic animals. Since in obesity adipokine plasmatic levels changes, in this study leptin (LEP) system was evaluated in the skin of obese dogs to observe changes in peripheral tissue. LEP is a hormone produced mainly by the adipose tissue and its serum level may reflect body mass index and BCS. LEP is also expressed in the skin and it has a prominent role in the biology of this tissue promoting cell proliferation and regulating the wound healing process. Investigation performed in obese and normal-weight dogs evidenced LEP and leptin receptor (LEPR) immunostaining in several skin structures. As LEP expression regards, differences were non-significant, while the LEPR transcripts appeared 10 fold higher in obesedogs. No differences were observed in the composition of skin associated immune system. The obese group-increased LEPR expression suggests that the receptor modulates the system control. The LEP system changes in the skin under obesity conditions however, the exact role of LEP in obese dog skin needs further insights. Abstract Obesity predisposes to several health problems including skin diseases. However, information on the relationship between obesity and skin disorders in pets is very scarce. Leptin (LEP) is mainly produced by adipose tissue and has a prominent role in skin biology. This study evaluated the LEP system in the skin of obese dogs compared to normal-weight animals. The investigation was carried out on 10 obese (Obese group) and 10 normal-weight (Normal-weight group) dogs through Real-time PCR and immunohistochemistry. Cells of skin associated immune system were also evaluated. No differences were evidenced between the two groups as well as skin inflammation. LEP differences were no significant, while LEPR transcript appeared 10-fold higher in obesedogs than in normal-weight ones. Immunostaining for both molecules was observed in several skin structures such as the epidermis, hair follicles, and glands. No differences appeared in the skin associated immune system composition. This study is a preliminary report showing that LEP system changes in obese dog skin. The increased LEPR expression observed in the obese group suggests that the receptor plays a modulating role in the system control. However, the exact role of LEPin the skin under obesity conditions needs further elucidation.

The aims of the current study were to describe presence and clinical role over time of Streptococcus pluranimalium isolated in milk samples of Mediterranean buffalo (MB). Two hundred composite milk samples originating from 40 primiparous MB were collected at 10, 30, 60, 90, and 150d in milk (DIM) and from 20 pluriparous MB at 77 to 120 DIM. Milk samples were used for analysis of somatic cell counts, bacteriological cultures, and identification (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry). Nine of 200 (4.5%) samples of primiparous MB and 3 of 20 (15%) samples of pluriparous MB were positive for Strep. pluranimalium. The prevalence of the bacterium in primipari was 0% (0/40) at 10, 30, and 150 DIM, whereas it was 5 (2/40) and 17.5% (7/40) at 60 and 90 DIM, respectively. Eight primipari were positive only once, whereas 1 was positive at 2 different samplings. Mono-infection was not detected in any of the age categories or udder health status. Infections were transient in primipari. Clinical mastitis was observed in primipari once at 90 DIM, subclinical mastitis detected twice in the same animals at 60 and 90 DIM, and intramammary infections were diagnosed 1 and 5 times at 60 and 90 DIM in primipari, respectively, whereas 3 infections were diagnosed in pluripari. The clinical reflections demonstrate for the first time the presence of Strep. pluranimalium in MB and its association with different udder health status. Nevertheless, it cannot be excluded that the bacterium may simply follow a pattern of commensal or opportunistic behavior, taking advantage of a preexisting bacterial udder infection.

The aim of this study was to investigate the repeat breeding condition in Italian Mediterranean buffaloes that failed to conceive after at least 300 days in milk. The trial was carried out on 40 pluriparous Italian Mediterranean buffaloes with more than 300 days in milk. All the animals underwent ultrasound examination to assess endometrial thickness and oestrus synchronization by the Ovsynch-TAI Program. On the day of oestrus, blood samples were collected for the haemocytometric cell count and biochemical assay, and the animals were slaughtered in a local abattoir. A post-mortem uterine flushing was performed using sterile saline for microbiological analyses. Furthermore, uterine biopsies were carried out for histopathological assessment. Finally, endometrial samples were used for real-time PCR (RT-PCR) analysis to evaluate the expression of genes involved in innate immune recognition of pathogens and the inflammatory response, such as Toll-like receptor (TLR)1, TLR8, interleukin (IL)-1β, IL-6, IL-8, COL4A2, connective tissue growth factor (CTGF), and cysteine-rich angiogenic inducer 61 (CYR61). Statistical analysis was performed by one-way ANOVA. Based on the infiltration of lymphocytes and plasma cells or endometrial gland, lymphatic, and blood vessel ectasia recorded in the histopathological examination, the animals were classified into three groups: healthy (H Group; n = 5), moderate endometritis (M Group; n = 25), and severe endometritis (S Group; n = 10). A significantly greater (P

The surgical treatment and follow-up of a bilateral fracture of the molar part of the horizontal mandibular ramus, of a dairy Mediterranean Buffalo in “the field” were described. The mandibular region was swollen, painful and unstable on palpation. The radiographic examination confirmed the diagnoses. The association between sedation and nerve block of the mandibulo-alveolar nerve assured restraint and analgesia. The fractures were stabilized by linear skeletal external fixation, recycled from human medicine allowing a good mouth occlusion. Fifty-five days after surgery the fractures were completely healed and the heifer regularly calved 3 months after without any consequence for calf health

Background In the last years the knowledges on Mediterranean Buffalo (MB) mastitis is remarkably improving, nevertheless the attention has been never focused on vaccination as preventive strategy for the control of mastitis in these ruminates. The aim of the current study was to assess clinical efficacy over time of two different preventive vaccination protocols against S. aureus mastitis, in primiparous MB.Vaccinated (VG) and not-vaccinated (N-VG) groups, of 30 MB each one, were selected from two different herds (herd A: VG1 and N-VG1; herd B: VG2 and N-VG2) of the same farm. Herd A received a double vaccination (Startvac®, 45 and 10 days before calving, protocol A), while in herd B an additional administration was performed (52 days after calving, protocol B). Bacteriological milk culture and assessment of somatic cell count (SCC) were performed at 10, 30, 60 and 90 days in milk (DIM) from composite milk samples. After 90 DIM, daily milk yields and SCC values were monthly detected until dry-off. Results The overall incidence of positive MB for S. aureus was 40.8% (49/120) in VG1 and 43.3% (52/120) in N-VG1 (Protocol A), while 45.8% (55/120) and 50.8% (61/120) in VG2 and N-VG2 (Protocol B). The latter was associated with a significant decreased in prevalence (at 90 DIM) and incidence of mastitis (animals positive for S. aureus, SCC > 200^103, with or without clinical signs) in the vaccinated MB, while no difference occurred in protocol A. Moreover, herd B showed a significant reduction in prevalence of intramammary infection (animals positive for S. aureus, SCC

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.

Background: Lameness represents the third most important health-related cause of economic loss in the dairy industry after fertility and mastitis. Although, dairy Mediterranean Buffaloes (MB) and dairy cows share similar breeding systems predisposing to similar herd problems, published studies exploring its relevance and role in these ruminants are still rare and incomplete. The aims of this study were to describe the clinical findings of foot disorders (FDs) in dairy MB and their influence on animal welfare, determined by assessment of locomotion score (LS), body condition score (BCS) and cleanliness score (CS). Results: Of 1297 multiparous MB submitted to routine trimming procedures, 229 buffaloes showed at least one FD. The prevalence of buffaloes affected by FDs was 17.7 %, while motility and lameness indexes were 84.1 % (1091/1297) and 15.9 % (206/1297), respectively. Overgrowth was present in 17.0 % (220/1297), corkscrew claw in 15.8 % (205/1297), interdigital phlegmon in 0.9 % (12/1297), white line abscess in 0.8 % (11/1297), digital dermatitis in 0.1 % (1/1297) and interdigital hyperplasia in 0.1 % (1/1297). Simultaneous presence of FDs was recorded in 17.0 % of MB (221/1297): overgrowth and corkscrew claw occurred together in 15.8 % of cases (205/1297), overgrowth and interdigital phlegmon in 0.3 % (4/1297), overgrowth and white line abscess in 0.8 % (11/1297), digital dermatitis and interdigital hyperplasia in 0.1 % (1/1297). The presence of FDs was always associated with lameness (LS > 2), except from 23 MB with simultaneous overgrowth and interdigital phlegmon occurrence. The majority of MB within the under-conditioned group (95.5 %, 43/45) and all those with CS > 2 (122/122) had a locomotion score above the threshold of normality (LS > 2). Furthermore, foot diseases such as interdigital hyperplasia, white line abscess and digital dermatitis or interdigital hyperplasia seemed to occur more frequently associated with decreased BCS and increased CS scores. Conclusions: This study describes for the first time the involvement of white line disease, interdigital phlegmona, digital dermatitis and interdigital hyperplasia in foot disorders of dairy Mediterranean buffalo and shows their association with an impairment of animal welfare. © 2016 The Author(s).

Forty-four dogs naturally infected by Leishmania infantum were divided into two groups: 20 thrombocytopenic dogs with fewer than 150 x 10(9) platelets/l, and 24 non-thrombocytopenic dogs with more than 200 x 10(9) platelets/l. Ten clinically healthy dogs were used as controls. A haematological profile was obtained and the dogs' serum was used to assess the presence of platelet-binding IgM and IgG antibodies using a flow cytometry technique. Nineteen of the 20 thrombocytopenic dogs, and 13 of the 24 non-thrombocytopenic dogs had detectable levels of platelet-binding immunoglobulins, but none of the control dogs did so. The differences were significantly different for both IgM and IgG platelet-binding antibodies.

Since it was first developed in South Africa, the FAMACHA system has been successfully tested in different geographic area where Haemonchus contortus is the major gastrointestinal (GI) strongyle species of sheep. The present study, the first on the use of the FAMACHA system in Italy, was aimed at obtaining a complete haematological profile in sheep with GI strongyle infection, in order to evaluate the validity of the FAMACHA system for detecting the severity of anaemia in sheep from southern Italy. Between April and September 2006, 793 blood and faecal samples were collected from 137 adult sheep of different local breeds of the Campania region (southern Italy). Each study animal was monthly subjected to blood collection, GI strongyle fecal egg counts (FEC) and a complete clinical examination, with particular attention to the colour of ocular mucousmembranes. The FAMACHA score values (from 1 to 5) were compared to Hematocrit (Hct) and Haemoglobin (Hb) values. Considering the FAMACHA score number ‘‘3’’ as borderline value for anaemia, the eye score values for anaemic animals were considered in the categories of 4 and 5 (Group 4-5) or of 3, 4 and 5 (Group 3-4-5), respectively. Anaemia due to GI strongyle infection was considered when the Hct and/or Hb values were under 27% and 9 g/dl, respectively. The correlation between FAMACHA score and both Hb and Hct was significant (P < 0.001) and negative; however, the low correlation coefficients indicated a weak negative association between FAMACHA and either Hb and Hct parameters. The sensitivity, specificity, predictive negative value (NPV) and predictive positive value (PPV) of the FAMACHA system were calculated. The highest values of sensitivity were 66% and 60% in the Group 3-4-5 for Hct and Hb, respectively. The highest values of specificity were 98% in the Group 4-5, for Hb and Hct, respectively. In the Group 3-4-5, the specificity was 60% and 62% for Hb and Hct, respectively. The NPV was 92% (Hct) and 84% (Hb) in the Group 3-4-5, while it was 87% (Hct) and 78% (Hb) in the Group 4-5. The PPV, for the others categories considered (Hb, Hct, Group 4-5 and 3-4-5), was always below 54%. In conclusion, in the present study, the FAMACHA system showed a low sensitivity in detecting anemic sheep; these findings indicate that in southern Italy, under our experimental conditions, FAMACHA may represent an additional element that can integrate clinical examinations. It is important that such evaluations are performed in each region before the system is implemented, as there could be local issues that affect the accuracy of the system.

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.

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. © 2016 Elsevier Ltd