Diagnostic predictivity of transtubular potassium gradient for primary hypoadrenocorticism in hyperkalemic dogs: a cross-sectional study.

OBJECTIVES: Transtubular potassium gradient has never been evaluated in the diagnosis of primary hypoadrenocorticism in dogs. The objective of this study is to evaluate the diagnostic utility of transtubular potassium gradient for the diagnosis of primary hypoadrenocorticism in dogs with serum K+ > 5.5 mol/L. MATERIALS AND METHODS: Cross-sectional study including dogs with primary hypoadrenocorticism and dogs with other disease. Differences in signalment and laboratory parameters between groups were assessed by binary logistic regression modelling. Variables significantly associated with primary hypoadrenocorticism were evaluated for their diagnostic accuracy of this condition by receiver operator characteristic curve analysis and by multivariable logistic regression analysis, respectively. RESULTS: Dogs with primary hypoadrenocorticism (n=39) were significantly younger than hyperkalemic dogs without primary hypoadrenocorticism (n=41). Moreover they had higher serum and urinary concentrations of Na+ and Na+ :K+ , but lower transtubular potassium gradient than hyperkalemic dogs without hypoadrenocorticism. Serum Na+ (sNa+ ) had the highest diagnostic accuracy (area under the curve [AUC]: 0.87, 95% confidence interval, 0.78 to 0.96), followed by the transtubular potassium gradient (AUC: 0.81, 0.72 to 0.91), however the accuracy of the two AUCs was not statistically different. The transtubular potassium gradient-sNa+ additive index (i.e. transtubular potassium gradient+sNa+ /10) had a greater accuracy (AUC: 0.88, 0.79 to 0.94) than transtubular potassium gradient or sNa+ , although it was not statistically superior to the latter. The multivariable logistic regression model showed that transtubular potassium gradient (adjusted odds ratio [aOR]: 0.51, 0.3 to 0.84), sNa+ (aOR: 0.86, 0.78 to 0.95) and spayed female (aOR: 24.96, 2.32 to 267.66) were associated with primary hypoadrenocorticism. CLINICAL SIGNIFICANCE: Transtubular potassium gradient-sNa+ additive index can contribute to decision-making on diagnosis of primary hypoadrenocorticism in dogs presenting with hyperkalaemia.

First detection of porcine circovirus type 2e in Europe.

Porcine circovirus 2 (PCV-2) infections are among the most economically important in swine farming. Because of the high evolutionary rate of PCV-2, several variants have emerged and are currently classified into several genotypes. However, only three (i.e. PCV-2a, PCV-2b, and PCV-2d) have a worldwide distribution, with other genotypes restricted to certain geographical regions and/or for limited time periods. Underdiagnosis or underreporting of these genotypes cannot be excluded. This is the first report of the detection and genetic characterisation of the PCV-2e genotype in Europe, from sows on a farm in Italy showing no clinical evidence of porcine circovirus disease. A follow-up study demonstrated persistent subclinical evidence of PCV-2e on the farm, at low frequency and viral load. This incidental finding emphasises the need for more intensive routine monitoring activities involving asymptomatic animals, coupled with DNA sequencing and data sharing. Considering the relevant genetic and phenotypic divergence of such genotypes, the actual efficacy of currently applied vaccines and diagnostic assays should be further evaluated.

Retrospective detection of Porcine circovirus 3 (PCV-3) in pig serum samples from Spain.

Porcine circovirus 3 (PCV-3) is an emerging circovirus species that has recently been reported in different countries around the world, suggesting a widespread circulation. In this study, sera samples originating from 654 pigs of different production phases and clinical/pathological conditions, submitted for diagnostic purposes between 1996 and 2017, were randomly selected. Detection of PCV-3 genome in such samples was attempted with a previously described PCR method, and the partial genome sequence was obtained from selected PCV-3-positive samples from different years. Compiled data confirmed that PCV-3 has been circulating in the Spanish pig population since 1996. The overall frequency of PCV-3 PCR-positive samples in the study period was 11.47% (75 of 654). Phylogenetic analysis of twelve PCV-3 partial sequences obtained showed a high nucleotide identity with the already known PCV-3 sequences, with minor variations among years. No significant correlation was found between the detection of PCV-3 and any production phase nor clinical/pathological condition. These results confirm PCV-3 circulation at least since 1996 in the Spanish pig population with a low/moderate frequency. Although the information obtained was limited, PCV-3 did not appear to be linked to any specific pathological condition or age group.

Automated computation of femoral angles in dogs from three-dimensional computed tomography reconstructions: Comparison with manual techniques.

The aim of this ex vivo study was to test a novel three-dimensional (3D) automated computer-aided design (CAD) method (aCAD) for the computation of femoral angles in dogs from 3D reconstructions of computed tomography (CT) images. The repeatability and reproducibility of three manual radiography, manual CT reconstructions and the aCAD method for the measurement of three femoral angles were evaluated: (1) anatomical lateral distal femoral angle (aLDFA); (2) femoral neck angle (FNA); and (3) femoral torsion angle (FTA). Femoral angles of 22 femurs obtained from 16 cadavers were measured by three blinded observers. Measurements were repeated three times by each observer for each diagnostic technique. Femoral angle measurements were analysed using a mixed effects linear model for repeated measures to determine the levels of intra-observer agreement (repeatability) and inter-observer agreement (reproducibility). Repeatability and reproducibility of measurements using the aCAD method were excellent (intra-class coefficients, ICCs≥0.98) for all three angles assessed. Manual radiography and CT exhibited excellent agreement for the aLDFA measurement (ICCs≥0.90). However, FNA repeatability and reproducibility were poor (ICCs<0.8), whereas FTA measurement showed slightly higher ICCs values, except for the radiographic reproducibility, which was poor (ICCs<0.8). The computation of the 3D aCAD method provided the highest repeatability and reproducibility among the tested methodologies.

Full-genome sequencing of porcine circovirus 3 field strains from Denmark, Italy and Spain demonstrates a high within-Europe genetic heterogeneity.

Porcine circovirus 3 (PCV3) is a new species of the Circovirus genus, which has recently been associated with different clinical syndromes. Its presence has been reported in different countries of North and South America, Asia and recently also Europe (Poland). However, different from the other continents, no European PCV3 sequence is currently available in public databases. There is a strong need of epidemiological data and full-genome sequences from Europe because of its relevance in the understanding of PCV3 molecular epidemiology and control. To fill this lack of information, samples collected in Denmark, Italy and Spain in 2016 and 2017 were screened for PCV3. Of the Danish samples, 36 of 38 the lymph nodes, six of 20 serum samples and two of 20 lung samples tested positive. Similarly, 10 of 29 lungs, 20 of 29 organ pools, six of 33 sera and one of eight nasal swabs tested PCV3 positive in Italy. Fourteen of 94 serum pools from seven of 14 Spanish farms were also positive. Despite the convenience nature of the sampling prevents any precise prevalence estimation, the preliminary screening of the data from three European countries confirmed a rather wide PCV3 distribution in Europe. Furthermore, the analysis of the six obtained complete European PCV3 genomes and their comparison with the public available sequences seems to support a remarkable worldwide PCV3 circulation. These results underline once more the urgency of more extensive epidemiological studies to refine the current knowledge on PCV3 evolution, transmission, spreading patterns and impact on pig health.

Evaluation of 793/B-like and Mass-like vaccine strain kinetics in experimental and field conditions by real-time RT-PCR quantification.

Infectious bronchitis virus (IBV) is a great economic burden both for productive losses and costs of the control strategies. Many different vaccination protocols are applied in the same region and even in consecutive cycles on the same farm in order to find the perfect balance between costs and benefits. In Northern Italy, the usual second vaccination is more and more often moved up to the chick's first d of life. The second strain administration together with the common Mass priming by spray at the hatchery allows saving money and time and reducing animal stress. The present work compared the different vaccine strains (Mass-like or B48, and 1/96) kinetics both in field conditions and in a 21-day-long experimental trial in broilers, monitoring the viral replication by upper respiratory tract swabbing and vaccine specific real time reverse transcription PCR (RT-PCR) quantification. In both field and experimental conditions, titers for all the vaccines showed an increasing trend in the first 2 wk and then a decrease, though still remaining detectable during the whole monitored period. IBV field strain and avian Metapneumovirus (aMPV) presence also was also investigated by RT-PCR and sequencing, and by multiplex real-time RT-PCR, respectively, revealing a consistency in the pathogen introduction timing at around 30 d, in correspondence with the vaccine titer's main decrease. These findings suggest the need for an accurate knowledge of live vaccine kinetics, whose replication can compete with the other pathogen one, providing additional protection to be added to what is conferred by the adaptive immune response.

Association Between Pancreatitis and Immune-mediated Haemolytic Anaemia in Cats: A Cross-sectional Study.

In most cases, the underlying causes of feline pancreatitis and feline immune-mediated haemolytic anaemia (IMHA) remain unknown. Following the observation that some cats present with both diseases concurrently, the aim of this cross-sectional study was to investigate if the two disorders were associated statistically in this species. A residency case log of all cases seen at the Small Animal Hospital, University of Glasgow, Glasgow, UK, between July 2004 and December 2007, was used as the database for this study and an association between these two disorders was assessed by Fisher's exact test. Of the 155 sick cats included in the study, nine were diagnosed at the time of presentation with pancreatitis (prevalence of 5.8%) and 11 with IMHA (prevalence of 7.1%). In three of these cats both conditions were present concurrently at the time of presentation (prevalence of 1.9%). A significant association between these two disease was found (Odds Risk 8.63, exact 95% confidence interval 1.15-49.31, P = 0.0178). Two possible biological explanations for the statistical association found between these two diseases are that one of these conditions may represent the cause for the onset of the other or that an underlying disorder of the immune system may concurrently target feline red blood cells and the exocrine pancreas.

A surveillance system for diseases of companion animals in the Veneto region (Italy).

Experts and international public health organisations stress the lack of surveillance systems for companion animal diseases and the need to implement such surveillance as a priority of the 'One Health' perspective. This paper presents the features of a system for the collection, analysis, interpretation and dissemination of data regarding the health status of pets in the Veneto region (Italy). The system involved the construction of a Web-based database containing the diagnoses of transmissible and non-transmissible diseases of dogs and cats made by veterinarians in their practices, hospitals, kennels and catteries. Each diagnosis constitutes a single record, also containing data on the identification of the individual animal and on several characteristics of epidemiological relevance. The World Health Organization (WHO) 10th revision of the International Classification of Diseases (ICD-10) for human diseases has been adapted to canine and feline diseases to standardise the diagnostic nomenclature. Software has been specifically created for online data entry and data management. The first results show that the main disorders were digestive (21%), dermatological (18%) and cardiovascular (11%) among 1,087 diagnostic records in dogs, and digestive (23%), dermatological (15%) and urinary (14%) among 289 diagnostic records in cats. The main causes of death are represented by cardiovascular (21%) and gastrointestinal (21%) diseases in dogs and by urinary (31%) disorders in cats. At present, no institutional surveillance system for companion animal health exists in Italy, and veterinarians joining this project and sharing the outcomes of their clinical activity are acting on a voluntary basis.

Aussi bien les experts que les organisations internationales oeuvrant dans le domaine de la santé publique soulignent l'absence de systèmes de surveillance dédiés aux maladies des animaux de compagnie, alors que la mise en place de cette surveillance constitue une priorité dans une perspective « Une seule santé ¼. Les auteurs décrivent les caractéristiques d'un système introduit en Vénétie (Italie) pour collecter, analyser, interpréter et diffuser des données sur la situation sanitaire des animaux de compagnie. Le système repose sur une base de données en ligne alimentée par les rapports de diagnostic sur les maladies transmissibles et non transmissibles des chiens et des chats établis par les vétérinaires dans leur cabinet, à l'hôpital ou dans les élevages ou pensions pour chiens et chats. Chaque diagnostic fait l'objet d'une notification spécifique où sont également consignées les données d'identification individuelle de l'animal et les caractéristiques pertinentes au plan épidémiologique. La classification internationale statistique des maladies (ICD­10) de l'Organisation mondiale de la santé (OMS), qui concerne les maladies humaines, a été adaptée aux maladies des chiens et des chats afin d'utiliser une nomenclature standardisée des diagnostics. Un logiciel spécifique a été créé pour la saisie en ligne des données et leur gestion. D'après les premiers résultats, les principales affections diagnostiquées étaient, chez le chien (sur 1 087 rapports de diagnostic), des maladies digestives (21 %), dermatologiques (18 %) et cardio-vasculaires (11 %) et, chez le chat (sur 289 rapports de diagnostic), des maladies digestives (23 %), dermatologiques (15 %) et urinaires (14 %). Les principales causes de mortalité étaient respectivement les maladies cardio-vasculaires (21 %) et gastro-intestinales (21 %) chez le chien et les maladies du système urinaire (31 %) chez le chat. À l'heure actuelle, aucun système institutionnel de surveillance n'est en place en Italie pour les animaux de compagnie, de sorte que les vétérinaires qui participent à ce projet et partagent leurs résultats cliniques le font sur une base volontaire.

Tanto especialistas como organizaciones internacionales dedicadas a temas de salud pública hacen hincapié en la ausencia de sistemas de vigilancia de las enfermedades de los animales de compañía y en la necesidad de instaurar tal vigilancia como elemento prioritario de los planteamientos de «Una sola salud¼. Los autores presentan las características de un sistema destinado a reunir, analizar, interpretar y difundir datos sobre el estado de salud de los animales de compañía en la región italiana del Veneto. Para instituir ese sistema se creó una base de datos en línea que centraliza información sobre los diagnósticos de enfermedades transmisibles y no transmisibles de perros y gatos realizados por veterinarios en el ejercicio de su labor en consultorios, hospitales y residencias caninas y felinas. Cada diagnóstico constituye un registro único, que también contiene datos sobre la identidad del animal en cuestión y sobre una serie de aspectos de importancia epidemiológica. Con objeto de normalizar la nomenclatura de diagnóstico se adaptó a las enfermedades caninas y felinas la Clasificación Internacional de Enfermedades, décima revisión (CIE­10), de la Organización Mundial de la Salud (OMS), que se aplica a las enfermedades humanas. También se crearon programas informáticos destinados específicamente a la introducción de datos en línea y a su gestión. Los primeros resultados muestran que los principales trastornos en los perros, de los 1.087 diagnósticos registrados, fueron los digestivos (21%), seguidos de los dermatológicos (18%) y los cardiovasculares (11%). En el caso de los gatos, con 289 diagnósticos registrados, las dolencias más importantes fueron las digestivas (23%), las dermatológicas (15%) y las urinarias (14%). En el perro, las principales causas de mortalidad fueron las enfermedades cardiovasculares (21%) y gastrointestinales (21%), y en el gato las patologías urinarias (31%). Actualmente no existe en Italia ningún sistema institucional de vigilancia de la salud de los animales de compañía, y los veterinarios que participan en este proyecto y comparten los resultados de su praxis clínica lo hacen con carácter voluntario.

Gamma and Deltacoronaviruses in quail and pheasants from Northern Italy1.

In view of the restricted knowledge on the diversity of coronaviruses in poultry other than chicken, this study aimed to investigate the genetic diversity of coronaviruses in quail, pheasant, and partridge from two regions of Northern Italy. To this end, pools of tracheal and cloacal swabs from European quail (Coturnix Coturnix) and intestinal tract from pheasants (Phasianus Colchicus) and partridge (Perdix Perdix) flocks, with or without enteric signs, were collected during 2015. Avian coronavirus (Gammacoronavirus) was detected in quail not vaccinated against Infectious Bronchitis Virus (IBV) and in pheasants vaccinated with an IBV Massachusetts serotype. Based on DNA sequences for the gene encoding the S protein, the avian coronaviruses detected in the quail and pheasant are related to the IBV 793B and Massachusetts types, respectively. However, RNA-dependent RNA polymerase (RdRp) analyses showed the susceptibility of quail also to Deltacoronaviruses, suggesting that quail and pheasant avian coronaviruses share spike genes identical to chicken IBV spike genes and quail might host Deltacoronavirus.

Hemostatic Findings in Ascitic Fluid: A Cross-Sectional Study in 70 Dogs.

BACKGROUND: Ascitic fluids of horses and humans have fibrinolytic activity, independent of the underlying mechanism of fluid formation. OBJECTIVE: To determine whether coagulation and fibrinogenolytic/fibrinolytic activity (ie, low fibrinogen and increased fibrin-fibrinogen degradation products [FDPs], D-dimer, or both) occur in all types of ascitic fluid in dogs. ANIMALS: A total of 70 client-owned dogs with ascites. METHODS: In this cross-sectional study, dogs were categorized based on the pathophysiology of fluid formation into 4 groups: transudates due to decreased osmotic pressure, transudates due to increased hydrostatic pressure, exudates, and hemorrhagic ascites. Fibrinogen, FDPs, and D-dimer concentrations were measured and then compared in both ascitic fluid and plasma. RESULTS: Ten dogs had transudates due to decreased colloid osmotic pressure, 18 had transudates due to increased hydrostatic pressure, 13 had exudates, and 29 had hemorrhagic ascites. Ascitic fibrinogen concentrations (n = 70) were significantly lower (median = 59 mg/dL; range: 59-122 mg/dL) than those in the plasma (median = 168 mg/dL, range: 59-879 mg/dL; P < .0001). Ascitic FDPs concentrations (n = 70) were significantly higher (<5 µg/mL: 3/70 dogs, ≥5 to <20 µg/mL: 11/70 dogs, ≥20 µg/mL: 56/70 dogs) than those in the plasma (<5 µg/mL: 17/70 dogs, ≥5 to <20 µg/mL: 28/70 dogs, ≥20 µg/mL: 25/70 dogs; P < .0001). Ascitic D-dimer concentrations (n = 70) were significantly higher (median = 3.98 µg/mL, range: 0.02-9.19) than those in the plasma (median = 0.11 µg/mL, range: 0.01-4.08; P < .0001). Analysis of the data for each of the 4 different types of ascites showed similar results to those of all the data analyzed together. CONCLUSIONS AND CLINICAL IMPORTANCE: Ascitic fluid of dogs has evidence of coagulation activation and fibrinogenolytic/fibrinolytic activity and that this phenomenon occurs independent of the underlying mechanism that leads to the formation of ascites.

X-Linked Hereditary Nephropathy in Navasota Dogs: Clinical Pathology, Morphology, and Gene Expression During Disease Progression.

X-linked hereditary nephropathy (XLHN) in Navasota dogs is a spontaneously occurring disease caused by a mutation resulting in defective production of type IV collagen and juvenile-onset renal failure. The study was aimed at examining the evolution of renal damage and the expression of selected molecules potentially involved in the pathogenesis of XLHN. Clinical data and renal samples were obtained in 10 XLHN male dogs and 5 controls at 4 (T0), 6 (T1), and 9 (T2) months of age. Glomerular and tubulointerstitial lesions were scored by light microscopy, and the expression of 21 molecules was investigated by quantitative real-time polymerase chain reaction with selected proteins evaluated by immunohistochemistry. No significant histologic lesions or clinicopathologic abnormalities were identified in controls at any time-point. XLHN dogs had progressive proteinuria starting at T0. At T1, XLHN dogs had a mesangioproliferative glomerulopathy with glomerular loss, tubular necrosis, and interstitial fibrosis. At T2, glomerular and tubulointerstitial lesions were more severe, particularly glomerular loss, interstitial fibrosis, and inflammation. At T0, transforming growth factor ß, connective tissue growth factor, and platelet-derived growth factor α mRNA were overexpressed in XLHN dogs compared with controls. Clusterin and TIMP1 transcripts were upregulated in later stages of the disease. Transforming growth factor ß, connective tissue growth factor, and platelet-derived growth factor α should be considered as key players in the initial events of XHLN. Clusterin and TIMP1 appear to be more associated with the progression rather than initiation of tubulointerstitial damage in chronic renal disease.

Subpopulations in aMPV vaccines are unlikely to be the only cause of reversion to virulence.

Avian metapneumovirus (aMPV) infects respiratory and reproductive tracts of domestic poultry, often involving secondary infections, and leads to serious economic losses in most parts of the world. While in general disease is effectively controlled by live vaccines, reversion to virulence of those vaccines has been demonstrated on several occasions. Consensus sequence mutations involved in the process have been identified in more than one instance. In one previous subtype A aMPV candidate vaccine study, small subpopulations were implicated. In the current study, the presence of subpopulations in a subtype B vaccine was investigated by deep sequencing. Of the 19 positions where vaccine (strain VCO3/50) and progenitor (strain VCO3/60616) consensus sequences differed, subpopulations were found to have sequence matching progenitor sequence in 4 positions. However none of these mutations occurred in a virulent revertant of that vaccine, thereby demonstrating that the majority progenitor virus population had not survived the attenuation process, hence was not obviously involved in any return to virulence. However within the vaccine, a single nucleotide variation was found which agreed with consensus sequence of a derived virulent revertant virus, hence this and other undetected, potentially virulent subpopulations, can be involved in reversion. Much deeper sequencing of progenitor, vaccine and revertant may clarify whether problematic virulent subpopulations are present and therefore whether these need to be routinely removed during aMPV vaccine preparation prior to registration and release.

A sensitive, reproducible, and economic real-time reverse transcription PCR detecting avian metapneumovirus subtypes A and B.

Use of real-time PCR is increasing in the diagnosis of infectious disease due to its sensitivity, specificity, and speed of detection. These characteristics make it particularly suited for the diagnosis of viral infections, like avian metapneumovirus (AMPV), for which effective control benefits from continuously updated knowledge of the epidemiological situation. Other real-time reverse transcription (RT)-PCRs have been published based on highly specific fluorescent dye-labeled probes, but they have high initial cost, complex validation, and a marked susceptibility to the genetic variability of their target sequence. With this in mind, we developed and validated a SYBR Green I-based quantitative RT-PCR for the detection of the two most prevalent AMPV subtypes (i.e., subtypes A and B). The assay demonstrated an analytical sensitivity comparable with that of a previously published real-time RT-PCR and the ability to detect RNA equivalent to approximately 0.5 infectious doses for both A and B subtypes. The high efficiency and linearity between viral titer and crossing point displayed for both subtypes make it suited for viral quantification. Optimization of reaction conditions and the implementation of melting curve analysis guaranteed the high specificity of the assay. The stable melting temperature difference between the two subtypes indicated the possibility of subtyping through melting temperature analysis. These characteristics make our assay a sensitive, specific, and rapid tool, enabling contemporaneous detection, quantification, and discrimination of AMPV subtype A and B.

Comparative field study to evaluate the performance of three different traps for collecting sand flies in northeastern Italy.

Three standard methods for collecting sand flies (sticky trap, CDC light trap, and CO2 trap) were compared in a field study conducted from June to October, 2012, at a site located in the center of a newly established autochthonous focus of canine leishmaniasis in northeastern Italy. Six traps (two sticky traps, two CDC light traps, and two CO2 traps) were activated at the same time for a single night every two weeks during the season of sand fly activity. A total of 5,667 sand flies were collected and 2,213 identified, of which 82.1% were Phlebotomus perniciosus, 17.4% P. neglectus, 0.3% Sergentomya minuta, and 0.2% P. mascitti. The performances of all traps were influenced by their position inside the site, increasing with proximity to the animal shelters. CO2 traps were more attractive for females of P. perniciosus and P. neglectus. CDC light traps showed an intermediate efficiency and were more attractive for P. neglectus, compared to other two traps. Results suggest that in northern Italy the CO2 trap is a suitable sampling method for sand fly monitoring programs that include transmitted pathogen surveillance.

Antimicrobial resistance in bacteria from breeding dogs housed in kennels with differing neonatal mortality and use of antibiotics.

This work examines the antimicrobial resistance of potentially pathogenic bacteria (Staphylococcus pseudintermedius, Streptococcus canis, Escherichia coli) found in the vaginal tract in prepartum mammary secretions and postpartum milk of bitches housed in breeding kennels (N = 20; 92 bitches). The kennels were divided into three categories: no routine antimicrobial administration around parturition (category 1); routine administration of one antibiotic around parturition (category 2); routine administration of multiple antimicrobials around parturition (category 3). Bacteriological cultures and antibiotic susceptibility tests were performed on vaginal specimens, prepartum mammary secretions, and postpartum milk. Stillbirths and neonatal deaths were recorded for each whelping and analyzed as "within-litter stillbirths" and "within-litter neonatal deaths" according to kennel category, by Pearson χ(2) test and the Kruskal-Wallis nonparametric test, respectively. The frequency of isolation and antimicrobial resistance of bacteria were analyzed according to kennel category by Pearson χ(2) test. Kennel category was not significantly associated with differing numbers of stillbirths or neonatal death events, nor was the frequency of isolation of potentially pathogenic bacteria in the three kennel categories significantly different. Kennel category 3 had a significantly higher frequency of isolation of multiresistant gram-positive bacterial strains. Our results show that intense administration of antibiotics to breeding bitches does not effectively reduce neonatal mortality; on the contrary, it induces multiresistance in potentially pathogenic bacteria. Breeders and veterinarians should be aware of the risk of selecting pathogenic bacteria by uncontrolled treatment in prepartum bitches.

Study on ticks and tick-borne zoonoses in public parks in Italy.

A survey on tick density and on tick-borne zoonoses was carried out in four public parks in the outskirts of Imola (northern Italy) from June to October 2006. All stages of Ixodes ricinus and only larvae of Riphicephalus sanguineus were recovered by dragging, performed on 100-m transects. Almost all ticks (99%) were harvested in one park. I. ricinus density (nymphs/100 m(2) ) ranged from 0 in park L to 6.3 in park F. Nymphs and adults of I. ricinus were subjected to PCR for Anaplasma phagocytophilum, Bartonella spp., Borrelia burgdorferi s. l. and Rickettsia spp. The observed prevalences were 38.3% for Bartonella henselae, 5.2% for Bartonella clarridgeiae, 10.4% for B. burgdorferi s. l., 2.6% for Rickettsia helvetica and 13% for Rickettsia monacensis, respectively. No DNA of A. phagocytophilum was found. Acarological risks (AR) were calculated as probabilities of collecting at least one infected nymph per transect. The AR values calculated for the various zoonotic agents were 11.4% for R. helvetica, 27.7% for B. clarridgeiae, 49.7% for B. burgdorferi s. l., 57.2% for R. monacensis and 90.4% for B. henselae, respectively. In this study, B. clarridgeiae was for the first time identified in I. ricinus ticks.

Cytauxzoon sp. infection in the first endemic focus described in domestic cats in Europe.

Information about epidemiological and clinicopathological aspects of domestic cat infection by species of Cytauxzoon other than Cytauxzoon felis is limited and it has rarely been reported. Following the detection of clinical cytauxzoonosis in three cats from Trieste (Italy), an epidemiological study was carried out in colony (n=63) and owned (n=52) cats from the same city to investigate the presence of Cytauxzoon sp. infection and to assess clinicopathological findings and variables associated with this infection. Cytauxzoon sp. infection was detected by 18S rRNA gene PCR in 23% (27/118) and by blood smear examination in 15% (18/118) of domestic cats. The 18S rRNA gene sequences obtained were 99% identical to the Cytauxzoon sp. sequences deposited in GenBank(®) from Spanish, French and Mongolian wild and domestic cats. Erythroparasitemia was observed mainly in apparently healthy cats. Cytauxzoon sp. infection was statistically associated with the colony group and the outdoor life style. No statistical association was found between positivity by PCR and breed, gender, age, presence of ticks and/or fleas, clinical status, laboratory findings such as anemia, FIV and/or FeLV status and mortality rate. Persistence of the infection was monitored and documented in four clinical cases. We reported the first clinicopathological description of naturally occurring Cytauxzoon sp. infection in domestic cats living in Italy. The predominance of subclinical erythroparasitemia and the evidence of persistent infection support the hypothesis that the domestic cat might serve as a reservoir host for this infection.

Immunohistochemical study of phospho-Stat3-ser727 expression in feline mammary gland tumours.

We describe the expression of pStat3-ser727 (signal transducer and activator of transcription 3 phosphorylated on serine 727) in normal, hyperplastic and neoplastic feline mammary gland tissue assessed by immunohistochemistry in 56 cats. The samples included 4 normal mammary non-lactating tissues, 13 hyperplastic lesions (9 lobular and 4 fibroepithelial) and 39 tumours (6 benign and 33 carcinomas). For immunohistochemistry, tissue sections were incubated with anti-pStat3-ser727 monoclonal antibodies and visualized with EnVision-DAB polymer. pStat3-ser727 positivity was quantified in a semi-quantitative manner, differentiating cytoplasmic and nuclear localization. Intense anti-pStat3-ser727 immunoreactivity was detected in epithelial neoplastic cells and in the fibro-component in two fibroepithelial hyperplastic lesions. The immunostaining was dot-like in the cytoplasm and homogeneous in the nuclei in both benign and malignant lesions. Statistically significant relations were observed between nuclear expression of pStat3-ser727 and the pleomorphism score (p = 0.006), mitotic activity (p < 0.0001), and histological grade (p < 0.0001). In contrast, no significant correlations were observed for cytoplasmic pStat3-ser727. These findings add new and interesting information on the potential role of the phosphorylated form of Stat3 in malignant lesions.

Proposal of Health Information System (HIS) as tool for the epidemiological surveillance of leishmaniasis in urban areas.

In recent years leishmaniasis has become a serious Public Health problem in Italy. The coexistence of infected dogs, vectors and humans, makes urban areas suitable environment for the spread of this zoonotic disease. With the purpose to improve the current system for the control of leishmaniasis in urban areas, the authors report their experiences carrying out a Health Information System (HIS) for the surveillance of the disease in the city of Messina. The HIS project is built on a dataset containing geographical data on urban environment, census tracts at parish level and data of human and canine leishmaniasis cases reported in the last years. The HIS allows the spatial representation of the disease occurrence and can be used as a novel epidemiological tool to perform area-based surveillance.