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The primary aim of this study was to assess whether human Light's criteria with the cut-off values previously published for cats are useful and superior to the traditional veterinary classification in diagnosing pathophysiology of fluid formation in cats with pleural effusion. The secondary aim was to assess if the albumin gradient (ALBg) is a reliable criterion for differentiating exudates from transudates in patients with pleural effusion thought to be transudative by clinical criteria but identified as exudative by Light's criteria. Nineteen client-owned cats with pleural effusion were studied. The aetiology of the pleural effusion was used to establish the pathophysiology of its formation. Parameters measured or calculated undergoing statistical analysis included Light's criteria, total protein and total nucleated cell count in the pleural effusions, and the ALBg. Based on the pathophysiology of fluid formation there were seven transudates caused by increased hydrostatic pressure and 12 exudates. There was a significant difference in the accuracy of the Light's criteria in correctly classifying origin of the pleural fluid formation compared with the traditional veterinary classification (84% vs 53%). ALBg values were significantly different between transudates and exudates. One of the three transudates misclassified as exudates by Light's criteria was correctly identified as a transudate by the ALBg. In conclusion, pleural effusion should be classified as either a transudate or an exudate using Light's criteria. In cats with pleural effusion thought to be transudative by clinical criteria, but identified as exudative by Light's criteria, the ALBg may further help in correctly differentiating exudates from transudates., (© The Author(s) 2015.)

Since the first description of Porcine circovirus type 2 (PCV2), four genotypes (PCV2a, PCV2b, PCV2c and PCV2d) have been recognized and three of them have been shown to exhibit worldwide distribution. Here, the population dynamics of PCV2 has been reconstructed over time and the factors that have shaped its evolution determined. The results obtained confirm that PCV2 originated approximately at the beginning of the 20th century. The most recent common ancestor of genotypes PCV2a, PCV2b, PCV2c and PCV2d circulated in the 1950s, 1980s, 1960s and 1950s, respectively, and the population sizes of the individual genotypes remained low until the mid 90s, coinciding with the identification of PCV2 as a major pathogen of the pig industry. The population dynamics of PCV2 have been characterized by the appearance of periodic waves of distinct genotypes that, after an initial rise, spread following major swine commercial routes and were then superseded by subsequent emerging genotypes. Various recombinant forms displayed comparable population dynamics and spreading routes to those of major genotypes, suggesting that recombinant strains are able to compete with parental ones. The capsid gene is subjected to immune selection and evasion of the host immune response seems to be a major force for the emergence and spread of new genotypes. In contrast, the evolution of other genes appears to be constrained by the particular genomic organization of PCV2. In summary, obtained results suggest that changes in farming strategies, international trade, host population immunity, recombination and the constraints imposed by genome organization have all played a major role in the evolutionary dynamics of PCV2., (Copyright © 2016 Elsevier Inc. All rights reserved.)

OBJECTIVE To determine the sensitivity, specificity, and interobserver variability of survey thoracic radiography (STR) for the detection of heart base masses (HBMs) in dogs. DESIGN Retrospective case-control study. ANIMALS 30 dogs with an HBM and 120 breed-matched control dogs (60 healthy dogs and 60 dogs with heart disease and no HBM). PROCEDURES In a blinded manner, 2 observers (designated as A and B) evaluated STR views from each dog for a mass-like opacity cranial to the heart, tracheal deviation, cardiomegaly, findings suggestive of pericardial effusion or right-sided congestive heart failure, and soft tissue opacities suggestive of pulmonary metastases. Investigators subsequently provided a final interpretation of each dog's HBM status (definitely affected, equivocal, or definitely not affected). RESULTS Considering equivocal interpretation as negative or positive for an HBM, the sensitivity of STR for diagnosis of an HBM was 40.0% (95% confidence interval [CI], 22.5% to 57.5%) and 56.7% (95% CI, 38.9% to 74.4%), respectively, for observer A and 63% (95% CI, 46.1% to 80.6%) and 80.0% (95% CI, 65.7% to 94.3%), respectively, for observer B. The corresponding specificity was 96.7% (95% CI, 93.5% to 99.9%) and 92.5% (95% CI, 87.8% to 97.2%), respectively, for observer A and 99.2% (95% CI, 97.5% to 100%) and 92.5% (95% CI, 87.8% to 97.2%), respectively, for observer B. The presence of a mass-like opacity cranial to the heart or tracheal deviation, or both, was significantly associated with a true diagnosis of HBM. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that STR is a highly specific but not a highly sensitive predictor of HBM in dogs.

Since its first description, PCV2 has emerged as one of the most economically relevant diseases for the swine industry. Despite the introduction of vaccines effective in controlling clinical syndromes, PCV2 spread was not prevented and some potential evidences of vaccine immuno escape have recently been reported ("Complete genome sequence of a novel porcine circovirus type 2b variant present in cases of vaccine failures in the United States" (Xiao and Halbur, 2012) [1], "Genetic and antigenic characterization of a newly emerging porcine circovirus type 2b mutant first isolated in cases of vaccine failure in Korea" (Seo et al., 2014) [2]). In this article, we used a collection of PCV2 full genomes, provided in the present manuscript, and several phylogentic, phylodynamic and bioinformatic methods to investigate different aspects of PCV2 epidemiology, history and evolution (more thoroughly described in "PHYLODYNAMIC ANALYSIS of PORCINE CIRCOVIRUS TYPE 2 REVEALS GLOBAL WAVES of EMERGING GENOTYPES and the CIRCULATION of RECOMBINANT FORMS"[3]). The methodological approaches used to consistently detect recombiantion events and estimate population dymanics and spreading patterns of rapidly evolving ssDNA viruses are herein reported. Programs used are described and original scripts have been provided. Ensembled databases used are also made available. These consist of a broad collection of complete genome sequences (i.e. 843 sequences; 63 complete genomes of PCV2a, 310 of PCV2b, 4 of PCV2c, 217 of PCV2d, 64 of CRF01, 140 of CRF02 and 45 of CRF03.), divided in differnt ORF (i.e. ORF1, ORF2 and intergenic regions), of PCV2 genotypes and major Circulating Recombinat Forms (CRF) properly annotated with respective collection data and country. Globally, all of these data can be used as a starting point for further studies and for classification purpose.

Since 1996 a new Infectious Bronchitis virus (IBV) genotype, referred to as Q1, circulated in China and was reported for the first time in Italy in 2011, associated with an increase of mortality, kidney lesions and proventriculitis. During northern Italian outbreak of respiratory disease in a broiler flock in 2013, an IBV strain was detected by RT-PCR and characterized as Q1-like based on partial S1 sequence. The virus was isolated and named γCoV/Ck/Italy/I2022/13. All coding regions of the isolate were sequenced and compared with 130 complete genome sequences of IBV and TCoV, downloaded from ViPR. This showed the highest identity with a Chinese strain CK/CH/LDL/97I (p-distance=0.044). To identify potential recombination events a complete genome SimPlot analysis was carried out which revealed the presence of possible multiple recombination events, but the minor parent strains remained unknown. A phylogenetic analysis of the complete S1 gene was performed using all complete S1 sequences available on ViPR and showed the isolate clustered with an Q1-like strain isolated in Italy in 2011 (p-distance=0.004) and a group of Chinese Q1-like strains isolated from the mid 90's (p-distance equal or higher than 0.001). It could be hypothesized that the isolate descended from the Italian 2011 Q1-like strain or was the result of a separate introduction from China through commercial trade or migratory birds; but the data currently available does not distinguish between these possibilities., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background: PCV2 has emerged as one of the most devastating viral infections of swine farming, causing a relevant economic impact due to direct losses and control strategies expenses. Epidemiological and experimental studies have evidenced that genetic diversity is potentially affecting the virulence of PVC2. The growing number of PCV2 complete genomes and partial sequences available at GenBank questioned the accepted PCV2 classification., Methods: Nine hundred seventy five PCV2 complete genomes and 1,270 ORF2 sequences available from GenBank were subjected to recombination, PASC and phylogenetic analyses and results were used for comparison with previous classification scheme., Results: The outcome of these analyses favors the recognition of four genotypes on the basis of ORF2 sequences, namely PCV2a, PCV2b, PCV2c and PCV2d-mPCV2b. To deal with the difficulty of founding an unambiguous classification and accounting the impossibility to define a p-distance cut-off, a set of reference sequences that could be used in further phylogenetic studies for PCV2 genotyping was established. Being aware that extensive phylogenetic analyses are time-consuming and often impracticable during routine diagnostic activity, ORF2 nucleotide positions adequately conserved in the reference sequences were identified and reported to allow a quick genotype differentiation., Conclusions: Globally, the present work provides an updated scenario of PCV2 genotypes distribution and, based on the limits of the previous classification criteria, proposes new rapid and effective schemes for differentiating the four defined PCV2 genotypes.

White striping ( WS: ) is an alteration of breast and thigh muscles of broiler chickens characterized by the presence of white striations parallel to the direction of muscle fibers. This study was performed to evaluate the prevalence and the predisposing factors to WS in commercial broilers of different weight reared in northern Italy. Fifty seven broiler flocks, including animals of medium- and heavy-weight, were grossly evaluated at slaughter for the presence of WS. For each flock, breeding data (mean BW at slaughter, ADG, sex, color of skin and fat, genetic line, age, antibiotic treatment, and prevalence of deep pectoral myopathy) were collected and statistically analyzed to assess their correlation with WS. Histology of breast fillets affected by different grades of WS was performed to evaluate potential differences between medium- and heavy-weight broilers. The overall prevalence of WS in medium- and heavy-weight broilers (mean BW 2.59 ± 0.13 kg and 3.64 ± 0.34 kg, respectively) was 70.2 ± 7.9% and 82.51 ± 8.5%, respectively, while the percentage of severe WS was 13.3 ± 7.1% and 25.7 ± 12.8%, respectively. A strong correlation was found between presence of WS, BW at slaughter, and ADG (Pearson correlation = 0.69, P < 0.01; Pearson correlation = 0.67, P < 0.01). WS also closely correlated with the prevalence of deep pectoral myopathy (Spearman's Rho slaughterhouse 1 = 0.74, Spearman's Rho slaughterhouse 2 = 0.51, P < 0.01). No correlation was found between genetics or sanitary status of the flock and WS. Histology confirmed that breasts with WS lesions were affected by a polyphasic degenerative and necrotizing myopathy, and that the lesions, as expected, were more severe in heavy-weight broilers. In conclusion, WS is a major concern in commercial meat poultry reared in Italy, affecting more severely heavier broilers, and it is mainly related to the BW and ADG of animals., (© 2015 Poultry Science Association Inc.)

Since its discovery, Porcine circovirus type 2 has emerged as one of the most relevant swine infectious diseases, causing relevant economic losses for the pig industry. While four genotypes were identified, only three (PCV2a, PCV2b and PCV2d) are currently circulating and display a worldwide distribution. Another genotype, PCV2c, has been described only once in Danish archive samples collected between 1980 and 1990. In addition to commercial pigs, PCV2 has been demonstrated to infect wild boars and other wild species, which can potentially serve as a reservoir for domestic populations. In this study, eight sequences obtained from feral pigs in the Pantanal region (Mato Grosso do Sul State, Brazil) were compared with reference sequences and other Brazilian sequences, and the results revealed remarkable genetic diversity, with all four genotypes currently recognised being detected (PCV2a, PCV2b, PCV2c and PCV2d). This finding represents a remarkable discovery, as it is the first detection of PCV2c since 1990 and the first-ever detection of PCV2c in live animals. The peculiar population history and ecological scenario of feral pigs in the Pantanal coupled with the complex, and still only partially known relationship of feral pigs with other PCV2 susceptible species (i.e., domestic pigs, wild boars and peccaries), open exciting questions concerning PCV2 origin and evolution. Overall, the results of the present study led us to form the following hypothesis: the PCV2 strains found in feral pigs may be the last descent of the strains that circulated among European pigs in the past, or they may have infected these feral pigs more recently through a bridge species., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Preureteral vena cava (circumcaval ureter, retrocaval ureter) occurs in a third of the feline population and has been associated with ureteral strictures in humans. The aim of this retrospective cross-sectional study was to describe the contrast-enhanced multidetector row computed tomographic (MDCT) characteristics of presumed preureteral vena cava in a group of cats. Medical records from two institutions located in different continents were searched from 2010-2013 for cases with complete contrast-enhanced MDCT examinations of the abdomen (i.e. included the entire course of the ureters and prerenal and renal segments of the caudal vena cava) and a diagnosis of preureteral caudal vena cava. For cases meeting inclusion criteria, CT scan data were retrieved and characteristics of the preureteral caudal vena cava were recorded. Presence of concomitant renal or ureteral diseases was also recorded. A total of 272 cats had contrast-enhanced abdominal CT scans during the study period and of these, 68 cats (22.43 ± 4.96%) had a diagnosis of presumed preureteral vena cava. In all affected cats, a "reverse-J ureter" was observed, i.e. a ureter running medially at the level of L4-5, passing dorsally to the caudal vena cava and then exiting ventrally between the caudal vena cava and aorta returning to its normal position. Having a preureteral vena cava resulted in an increased risk for concurrent urinary signs (OR = 3.00; CI: 95%; 1.28-6.99; P = 0.01). Findings supported the use of contrast-enhanced MDCT for characterizing morphology of preureteral vena cava and its relation with ureters in cats., (© 2015 American College of Veterinary Radiology.)

Porcine circovirus type 2 is one of the most widespread and economically relevant infections of swine. Four genotypes have been recognized, but currently, only three (PCV2a, PCV2b and PCV2d) are effectively circulating. The widespread livestock trade and rapid viral evolution have contributed to determining the high heterogeneity of PCV2 and the dispersal of potentially more virulent strains. Italian swine farming and the related processing industry are relevant in the national economy. Despite the noteworthy losses associated with direct and control measure costs, no data are currently available on the molecular epidemiology of PCV2 in Italy. Our study, which was intended to fill this gap, considered 75 completed genome PCV2 sequences, which were obtained from samples collected from the highly densely populated area of Northern Italy between 2007 and 2014. Phylogenetic analysis and comparison with reference sequences demonstrated the co-circulation, with different prevalences, of PCV2a, PCV2b and PCV2d within the national borders, with PCV2b being the most prevalent. Recombination between different genotypes was also proven to be frequent. Phylogeographic analysis demonstrated that the marked variability of Italian PCV2 strains can be attributable to multiple introduction events. The comparison of the phylogenetic analysis results, the location of different haplotypes and the international commercial routs of live pigs allow the speculation of several links as well as the role of Italy as both an importer and exporter of PCV2 haplotypes, mainly from and to European and Asian countries. A similarly intricate contact network was demonstrated within national borders, with different haplotypes being detected in the same province and different provinces harbouring the same haplotype. Overall, this paper represents the first description of PCV2 in Italy and demonstrates that the high variability of circulating Italian strains is due to multiple introduction events, wide circulation within national boundaries and rapid viral evolution., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Porcine reproductive and respiratory syndrome (PRRS) is the most relevant and challenging infectious disease to affect swine breeding. Despite this, several aspects of the virus' evolution and virus-host interaction are still poorly understood and largely based on knowledge obtained through in vitro or in vivo experimental infections. Due to peculiar experimental conditions, our understanding is often contradictory and difficult to infer with respect to actual field conditions. Our phylodynamic study, based on ORF5 sequences of 141 samples collected in Italy from 1993 to 2012, explores different aspects of PRRSV epidemiology, evolution, and virus-host interaction. Two major clades, belonging to Type 1 subtype 1, were demonstrated to co-circulate while harboring a relevant intra- and inter-clade genetic diversity. Most Recent Common Ancestor (MRCA), evolution rates, and population dynamics were estimated using a serial coalescent-based approach, and different demographic histories were reconstructed for the two clades. Analysis of selective pressure revealed that sites subjected to diversifying selection were mainly located in the region of glycoprotein 5 (GP5) exposed to the host environment. Similarly, the vast majority of strains were highly glycosylated, confirming the proposed protective role of the glycan shield against the humoral immune response. Overall, our study reports both interactions among the viral populations as well as between virus and host, and their relevance in shaping viral evolution: different population dynamics over time seem to reflect a competition between clades. Some evidence argues in favor of the role of immune pressure in affecting GP5 evolution, including frequent changes in the region exposed to the host immune response, and preserving glycosylation profiles that can hamper humoral immunity., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Recombination in Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is a well-documented phenomenon. A high recombination frequency has been reported in experimental conditions both in vitro and in vivo, and its role in driving viral evolution has been postulated by several authors. However field evidences are rare, mainly obtained from large-scale sampling and typically represented by single sequences rather than by groups of circulating "recombinant progenies". The present work was aimed to investigate the gray area between experimental studies and large-scale epidemiological investigations. The study was performed on ORF5, ORF7 and concatenated sequences obtained in our laboratory or available in GenBank collected between 2009 and 2012 in northern Italy. Six independent recombinant strains out of 66 concatenated sequences (∼9%) were found, demonstrating a high recombination frequency respect to previous field studies but comparable to in vitro experiments. In silico analysis let speculate that this new strain displayed physicochemical features diverse enough to potentially alter its immunological properties. Taken altogether, the results of our study support previous experimental evidences that depict PRRSV to be extremely prone to recombination. The limited temporal and geographical spread of recombinant strains however states in favor of a limited fitness of the recombinant progeny compared to parental strains and the marginal role of this phenomenon in PRRSV evolution., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Over a period of almost two years, broilers chickens on several hundred Italian farms, were monitored for infectious bronchitis virus. Detections were genotyped using a hypervariable region of the gene coding for the S1 segment of the spike protein. A range of genotypes were detected which comprised QX, Q1, Mass, D274 and 793B. Sequences of 793B viruses detected in chickens, vaccinated with either of the two commonly used 793B type vaccines were almost identical to sequences of one or other of these vaccines. This strong indication of vaccine association led to the withdrawal of live 793B vaccine use on all of the farms of the study. Except for one sample collected soon after 793B vaccination ceased, it was no longer possible to detect 793B vaccine on these farms. It appears that field 793B strains have disappeared from the region of Italy tested thus obviating any need for current vaccine protection against 793B., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

With respect to the epidemiology of leishmaniasis, it is crucial to take into account the ecoclimatic and environmental characteristics that influence the distribution patterns of the vector sand fly species. It is also important to consider the possible impact of on-going climate changes on the emergence of this disease. In order to map the potential distribution of Phlebotomus perniciosus, the main vector species of canine leishmaniasis in north-eastern Italy, geographical information systems tools, ecological niche models (ENM) and remotely sensed environmental data were applied for a retrospective analysis of an entomological survey conducted in north-eastern Italy over 12 years. Sand fly trapping was conducted from 2001 to 2012 in 175 sites in the provinces of Veneto, Friuli-Venezia Giulia and Trentino-Alto Adige. We developed a predictive model of potential distribution of P. perniciosus using the maximum entropy algorithm software, based on seasonal normalized difference vegetation index, day and night land surface temperature, the Corine land cover 2006, a digital elevation model (GTOPO30) and climate layers obtained from the WorldClim database. The MaxEnt prediction found the more suitable habitat for P. perniciosus to be hilly areas (100-300 m above the mean sea level) characterised by temperate climate during the winter and summer seasons, high winter vegetation cover and moderate rainfall during the activity season of vector sand fly. ENM provided a greater understanding of the geographical distribution and ecological requirements of P. perniciosus in the study area, which can be applied for the development of future surveillance strategies.

Caudal vena cava duplication has been rarely reported in small animals. The purpose of this retrospective study was to describe characteristics of duplicated caudal vena cava in a large group of dogs. Computed tomography (CT) and ultrasound databases from two hospitals were searched for canine reports having the diagnosis "double caudal vena cava." One observer reviewed CT images for 71 dogs and two observers reviewed ultrasound images for 21 dogs. In all CT cases, the duplication comprised two vessels that were bilaterally symmetrical and approximately the same calibre (similar to Type I complete duplication in humans). In all ultrasound cases, the duplicated caudal vena cava appeared as a distinct vessel running on the left side of the abdominal segment of the descending aorta and extending from the left common iliac vein to the left renal vein. The prevalence of caudal vena cava duplication was 0.46% for canine ultrasound studies and 2.08% for canine CT studies performed at these hospitals. Median body weight for affected dogs was significantly lower than that of unaffected dogs (P < 0.0001). Breeds with increased risk for duplicated caudal vena cava were Yorkshire Terrier (odds ratio [OR] = 6.41), Poodle (OR = 7.46), West Highland White Terrier (OR = 6.33), and Maltese (OR = 3.87). Presence of a duplicated caudal vena cava was significantly associated with presence of extrahepatic portosystemic shunt(s) (P < 0.004). While uncommon in dogs, caudal vena cava duplication should be differentiated from other vascular anomalies when planning surgeries and for avoiding misdiagnoses., (© 2014 American College of Veterinary Radiology.)

Although avian metapneumovirus (aMPV) infection has been reported in most regions of the world, to date, only subtype B has been detected in Egypt. At the end of November 2013, dry oropharyngeal swabs were collected during an outbreak of respiratory diseases in a free-range, multi-age turkey dealer farm in Northern Upper Egypt. The clinical signs that appeared when turkeys were 3 weeks-old were characterized by ocular and nasal discharge and swelling of sinuses. aMPV of subtype A was detected by real-time reverse transcription-polymerase chain reaction. In order to confirm the results and obtain more information on the molecular characteristics of the virus, F and G protein genes were partially sequenced and compared with previously published sequences deposited in GenBank by using BLAST. Subtype of the strain was confirmed by sequencing of partial F and G protein genes. The highest percentages of identity were observed when G sequence of the Egyptian strain was compared with the sequence of an aMPV-A isolated in Nigeria (96.4 %) and when the F sequence was compared with strains isolated respectively in Italy and in UK (97.1 %). Moreover, the alignment of the sequences with commercial subtype A vaccine or vaccine-derived strains showed differences in the Egyptian strain that indicate its probable field origin. The detection of aMPV in the investigated turkey flock highlights some relevant epidemiological issues regarding the role that multi-age farms and dealers may play in perpetuating aMPV infection within and among farms. To our knowledge, this is the first report of aMPV subtype A in Egypt.

The remarkable economic losses due to porcine reproductive and respiratory syndrome (PRRS) have stated the control and eradication of this disease is one of the main issues of swine modern farming. The limited cross-protection of vaccine-induced immunity compelled the adoption of strict biosecurity measures that must be associated with the prompt diagnosis of infection. In our study four RT-PCR methods, a RT-PCR, a SYBR Green I and two hydrolysis probes, were compared to evaluate their respective benefits and disadvantages. One hundred and seventy samples originating from 50 farms located in northern Italy were tested with all assays and performances were evaluated using a Bayesian approach to deal with the absence of a Gold Standard. Sequencing the complete of ORF7, the segment targeted by all methods, allowed a gain of insight into the genetic variability of Italian strains and to investigate the role of mismatches on assay sensitivity. Our study evidenced that methods based only on primers-genome interaction better tolerate PRRSV genetic variability, demonstrating a greater sensitivity (Se): SYBR Green I (Se=98.4%) and RT-PCR (Se=99%) outperform both in-house (Se=71.4%) and commercial (Se=91.7%) probe-based methods. On the other hand, probe-based assays allowed an easier genotyping of PRRSV strains and implementation of the internal control system (IC). Phylogenetic analysis allowed demonstration of a presence of two clades circulating continuously in northern Italy since 1996, when their probable ancestors were collected., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The accuracy and rapid diagnosis of PRRSV infection is a major prerequisite for every control and/or eradication strategy. In this study two real time RT-PCR based on different chemistry analysis (TaqMan Probes and SYBR Green) have been developed and validated before comparison to an end point two-step RT-PCR validated previously. All assays were aimed at discrimination between PRRSV genotypes. Furthermore, an exogenous internal control (IC) system had also been implemented in qRT-PCR. A rigorous analytical validation, executed on infected cell cultures and serum, demonstrated good sensitivity, specificity and repeatability. In particular RT-PCR was exceptionally sensitive and could detect a viral titre in the order of a magnitude of 1 copies/μL, 10-fold lower than other qRT-PCR described in this study. Optimal diagnostic performances have been demonstrated analyzing samples retrieved from an experimental infection, with RT-PCR again outperforming real time RT-PCR assays. All tests, showing substantial agreement between them, were able to detect early stages of viraemia (1 DPI) and some animals were classified as positive until the end of the study (76 DPI). Therefore, this supports the assays usefulness in animals with different clinical conditions and in a broad range of epidemiological scenarios. The benefits and disadvantages of different assays were also considered and discussed., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The most promising techniques for detecting minimal residual disease (MRD) in canine lymphoma are flow cytometry (FC) and polymerase chain reaction amplification of antigen receptor genes (PARR). However, the agreement between these methods has not been established. MRD was monitored by FC and PARR following treatment of dogs affected with diffuse large B-cell lymphoma (DLBCL), comparing results in lymph node (LN), peripheral blood (PB) and bone marrow (BM) samples. The prognostic impact of MRD on time to relapse (TTR) and lymphoma-specific survival (LSS) was also assessed. Fourteen dogs with previously untreated DLBCL were enrolled into the study; 10 dogs eventually relapsed, while four dogs with undetectable MRD were still in remission at the end of the study. At diagnosis, the concordance rate between FC and PARR was 100%, 78.6%, and 64.3% for LN, PB and BM, respectively. At the end of treatment, the agreement rates were 35.7%, 50%, and 57.1% for LN, PB and BM, respectively. At least one of the follow-up samples from dogs experiencing relapse was PARR(+); conversely, FC was not able to detect MRD in seven of the dogs that relapsed. PARR was more sensitive than FC in predicting TTR, whereas the combination of PARR and FC was more sensitive than either technique alone in predicting LSS using PB samples. The results suggest that immunological and molecular techniques should be used in combination when monitoring for MRD in canine DLBCL., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Objective: To evaluate RBC distribution width (RDW) in dogs with chronic degenerative valvular disease (CDVD) with compensated or decompensated heart failure., Design: Retrospective case-control study., Animals: 27 healthy dogs and 135 dogs with CDVD (87 dogs with compensated heart failure and 48 dogs with decompensated heart failure)., Procedures: The RDW and various CBC and serum biochemical variables were compared among groups. Correlations between RDW and various echocardiographic variables were evaluated., Results: Mean ± SD RDW in dogs with CDVD (13.1% ± 1.0%) was not significantly different from that of healthy dogs (12.8% ± 0.8%). The RDW of dogs with CDVD and compensated heart failure (13.0% ± 1.0%) was not significantly different from that of dogs with CDVD and decompensated heart failure (13.2% ± 1.1%). The RDW had a significant, weak, negative correlation with Hct (correlation coefficient, -0.250), hemoglobin concentration (correlation coefficient, -0.219), and mean corpuscular volume (correlation coefficient, -0.211). The RDW had a significant, weak, positive correlation with 1 echocardiographic index of CDVD severity (ie, the left atrium-to-aorta ratio [correlation coefficient, 0.183])., Conclusions and Clinical Relevance: In this study population, RDW did not seem to be associated with the presence of heart failure or CDVD.

Background: To evaluate the efficiency of platelet-rich plasma preparations by means of a double centrifugation tube method to obtain platelet-rich canine plasma at a concentration at least 4 times higher than the baseline value and a concentration of white blood cells not exceeding twice the reference range. A complete blood count was carried out for each sample and each concentrate. Whole blood samples were collected from 12 clinically healthy dogs (consenting blood donors). Blood was processed by a double centrifugation tube method to obtain platelet concentrates, which were then analyzed by a flow cytometry haematology system for haemogram. Platelet concentration and white blood cell count were determined in all samples., Results: Platelet concentration at least 4 times higher than the baseline value and a white blood cell count not exceeding twice the reference range were obtained respectively in 10 cases out of 12 (83.3%) and 11 cases out of 12 (91.6%)., Conclusions: This double centrifugation tube method is a relatively simple and inexpensive method for obtaining platelet-rich canine plasma, potentially available for therapeutic use to improve the healing process.

In recent years, special attention has been paid to real-time polymerase chain reaction (PCR) for avian metapneumovirus (AMPV) diagnosis, due to its numerous advantages over classical PCR. A new multiplex quantitative real-time reverse transcription-PCR (qRT-PCR) with molecular beacon probe assay, designed to target the SH gene, was developed. The test was evaluated in terms of specificity, sensitivity and repeatability, and compared with conventional RT nested-PCR based on the G gene. All of the AMPV subtype A and B strains tested were amplified and specifically detected while no amplification occurred with other non-target bird respiratory pathogens. The detection limit of the assay was 10(-0.41) median infectious dose/ml and 10(1.15) median infectious dose/ml when the AMPV-B strain IT/Ty/B/Vr240/87 and the AMPV-A strain IT/Ty/A/259-01/03 were used, respectively, as templates. In all cases, the amplification efficiency was approximately 2 and the error values were <0.2. Standard curves, generated either using the serial dilution of an RNA suspension or RNA extracted from the serial dilution of titrated viral suspensions as templates, exhibited good linearity (R (2)>0.9375) between crossing point values and virus quantities, making the assay herein designed reliable for quantification. When the newly developed qRT-PCR was compared with a conventional RT nested-PCR, it showed greater sensitivity with RNA extracted from both positive controls and from experimentally infected birds. This assay can be effectively used for the detection, identification, differentiation and quantitation of AMPV subtype A or subtype B to assist in disease diagnosis and to carry out rapid surveillance with high levels of sensitivity and specificity.

Objective: To determine whether dogs with ascites secondary to right-sided congestive heart failure (CHF) have bleeding disorders associated with hypofibrinogenemia and discordant plasma fibrin-fibrinogen degradation products (FDPs) and D-dimer assay results (ie, a circulating concentration of FDPs higher than the reference range and a circulating concentration of D-dimer within the reference range)., Design: Retrospective case-control study., Animals: 80 client-owned dogs., Procedures: Dogs with ascites secondary to right-sided CHF (group 1; n = 20), unhealthy dogs without cardiac disease (group 2; 40), and dogs with left-sided CHF (group 3; 20) were included in the study. Urine bile acids-to-creatinine concentration ratios were calculated as a marker of liver function. Differences among groups regarding coagulation profile analysis results and prevalence of discordant FDPs and D-dimer assay results were determined., Results: No significant differences were detected among the 3 groups regarding urine bile acids-to-creatinine concentration ratios. Plasma fibrinogen concentration was significantly lower for group 1 versus groups 2 or 3. Prevalence of discordant FDPs and D-dimer assay results was significantly higher for group 1 versus groups 2 or 3. Eighteen group 1 dogs had discordant FDPs and D-dimer assay results. Ten of these dogs had concurrent hypofibrinogenemia, 2 of which had clinical signs of bleeding. Only 10 dogs in groups 2 or 3 had discordant FDPs and D-dimer assay results; none of these dogs had hypofibrinogenemia or clinical signs of bleeding., Conclusions and Clinical Relevance: Dogs with right-sided CHF and ascites may be at increased risk for primary hyperfibrinogenolysis (ie, hypofibrinogenemia and discordant FDPs and D-dimer assay results).

Background: Cytologic diagnosis of hepatocellular carcinoma is possible when hepatocytes have prominent criteria of malignancy, but is problematic when hepatocytes have a relatively normal morphologic appearance., Objectives: The aim of this study was to determine the cytologic features of histologically confirmed well-differentiated hepatocellular carcinomas (WD-HCC) in dogs to develop reasonable criteria for an accurate diagnosis when obvious criteria of malignancy in hepatocytes are lacking., Methods: Slides of ultrasound-guided fine-needle aspirates of 15 histologically confirmed WD-HCC in dogs were evaluated retrospectively using a scoring system (0-3) established for 33 cytologic features. For comparison, 15 cytologic samples of non-nodular non-neoplastic liver were evaluated using the same criteria. Statistical analysis included multiple hypothesis testing using the Benjamini-Hochberg method or false discovery rate control to correct for multiple comparisons., Results: The most significant and useful cytologic features in the diagnosis of canine WD-HCC were dissociation of hepatocytes, acinar or palisading arrangements of neoplastic cells, and the presence of naked nuclei and capillaries, together with mild anisocytosis, anisokaryosis, multinuclearity, and increased N:C ratios., Conclusions: Cytologic features of canine WD-HCC were determined. Further studies are warranted to compare these cytologic criteria among WD-HCC, hepatomas, and hyperplastic nodules and to examine co-variance of the 33 cytologic features., (© 2012 American Society for Veterinary Clinical Pathology.)

The overuse of antimicrobials can select resistant bacteria strains; staphylococci have the ability to become resistant to all beta-lactam antimicrobials and are a significant concern in human medicine and a growing issue for veterinary medicine. Because antimicrobials are sometimes incorrectly used in breeding kennels, the objective of the work was to assess the occurrence of methicillin-resistant coagulase-positive staphylococci in breeding dogs. The research was carried out in 13 kennels that were allotted to three categories according to the intensity of antimicrobial use. Vaginal and milk swabs were taken from 87 healthy bitches around parturition and also from multiple organs of 27 of their pups that died within the first 2 weeks. Standard bacteriological examinations were carried out and coagulase-positive staphylococci were identified. All the coagulase-positive staphylococci resulted to be Staphylococcus pseudintermedius. Susceptibility to oxacillin and the presence of the mecA gene were tested. Nine out of 89 strains (six isolated from the bitches' milk and three from dead puppies, all belonging to kennels characterized by an excessive use of antimicrobials) were multidrug-resistant, methicillin-resistant and mecA positive. Our results confirm that excessive use of antimicrobials entails the risk of selecting resistant staphylococci strains. Our data also indicate that the bacterial flora of healthy dogs belonging to specific populations may act as a reservoir of resistance genes., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Objective: To detect abnormalities of the lower respiratory tract (trachea, principal bronchi, and lobar bronchi) in brachycephalic dogs by use of endoscopy, evaluate the correlation between laryngeal collapse and bronchial abnormalities, and determine whether dogs with bronchial abnormalities have a less favorable postsurgical long-term outcome following correction of brachycephalic syndrome., Design: Prospective case series study., Animals: 40 client-owned brachycephalic dogs with stertorous breathing and clinical signs of respiratory distress., Procedures: Brachycephalic dogs anesthetized for pharyngoscopy and laryngoscopy between January 2007 and June 2008 underwent flexible bronchoscopy for systematic evaluation of the principal and lobar bronchi. For dogs that underwent surgical correction of any component of brachycephalic syndrome, owners rated surgical outcome during a follow-up telephone survey. Correlation between laryngeal collapse and bronchial abnormalities and association between bronchial abnormalities and long-term outcome were assessed., Results: Pugs (n = 20), English Bulldogs (13), and French Bulldogs (7) were affected. A fixed bronchial collapse was recognized in 35 of 40 dogs with a total of 94 bronchial stenoses. Abnormalities were irregularly distributed between hemithoraces; 15 of 94 bronchial abnormalities were detected in the right bronchial system, and 79 of 94 were detected in the left. The left cranial bronchus was the most commonly affected structure, and Pugs were the most severely affected breed. Laryngeal collapse was significantly correlated with severe bronchial collapse; no significant correlation was found between severity of bronchial abnormalities and postsurgical outcome., Conclusions and Clinical Relevance: Bronchial collapse was a common finding in brachycephalic dogs, and long-term postsurgical outcome was not affected by bronchial stenosis.

We conducted a retrospective study to determine whether multidetector computed tomography (CT) could be of value for adrenal gland assessment in dogs with pituitary-dependent hyperadrenocorticism. Adrenal gland attenuation and volume values of 49 dogs with hyperadrenocorticism were recorded and age, body weight, and gender were examined to determine if a relationship existed between these variables and adrenal gland morphology. There was not a statistically significant difference in mean X-ray attenuation of the left vs. right adrenal gland in normal dogs (35.3 +/- 6.1 HU), or in dogs with hyperadrenocorticism. The mean adrenal X-ray attenuation (+/- standard deviation [SD]) in dogs with microadenoma was 33.1 +/- 6.8 vs. 31.8 +/- 12.7 HU for dogs with macroadenoma, and these values were not statistically different. The mean volume of the left adrenal gland in normal dogs (0.59 +/- 0.17 cm3) was greater than that of the right adrenal gland (0.54 +/- 0.19 cm3) (P < 0.05). The mean CT volume (+/- SD) of the adrenal glands in dogs with microadenoma vs. macroadenoma were 1.60 +/- 1.25 vs. 2.88 +/- 1.60 cm3, respectively. There was no effect of age or gender on adrenal gland morphology or X-ray attenuation. The weight effect was the most important source of variation for the volume measurement in dogs with hyperadrenocorticism.

STAT3 (signal transducer and activator of transcription 3) is a cytoplasmic transcription factor that plays a role in the G1 to S phase cell-cycle transition and is induced by cytokines and growth factors. The expression of STAT3 phosphorylated on tyrosine 705 (STAT3-p-tyr705) in normal, hyperplastic and neoplastic feline mammary gland tissue was assessed by immunohistochemistry in 45 cats. The samples included 4 normal mammary non-lactating tissues, 9 hyperplastic tissues (5 fibroepithelial hyperplasia and 4 lobular epithelial hyperplasia), 2 benign tumours (1 complex adenoma, and 1 simple adenoma), and 30 carcinomas (18 simple tubular papillary, 6 simple tubular, 2 simple solid, 3 cribriform, and 1 adenosquamous carcinoma). For immunohistochemistry, tissue sections were incubated with an anti-STAT3-p-tyr705 monoclonal antibody and visualized with EnVision-DAB polymer. STAT3-p-tyr705 positivity was quantified in a semi-quantitative manner. All positive samples showed cytoplasmic and/or nuclear positivity. Normal non-lactating mammary tissue showed a low number of positive cells, similar to hyperplastic tissue. In neoplastic tissues, a high number of positive cells with a moderate to intense reaction was observed. Moreover, a correlation was observed between nuclear positivity for STAT3-p-tyr705 and histologic grade (P=0.013; r=0.447), tubular formation (P=0.043; r=0.820), and mitotic activity (P<0.0001; r=0.689). In contrast, no such correlations were observed for cytoplasmic reactivity of STAT3-p-tyr705. A significant difference was observed between malignant lesions and hyperplasia with regards to expression of STAT3-p-tyr 705 in the cytoplasm (P=0.008; U=59.00) and nuclei (P=0.002; U=47.00). These results confirm previous our data and reinforce the potential role of STAT3 in malignancy as reported for human breast cancer and other sporadic tumours.

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.

The aim of this study was to examine the expression of signal transducer and activator of transcription 3 (STAT3) in canine haemangiomas and haemangiosarcomas. STAT3 protein was demonstrated in both the cytoplasm and nuclei of labelled cells. The expression of STAT3 was observed in 87.50% (35/40) of both haemangiomas and haemangiosarcomas and 72.22% (13/18) of haemangiomas and 100% (22/22) haemangiosarcomas. The percentage of positive cells in haemangiomas and haemangiosarcomas was statistically significant difference (P<0.0001). These preliminary results indicate that STAT3 is overexpressed in malignant tumours.

Geographic information systems (GIS) and remote sensing (RS) technologies are being used increasingly to study the spatial and temporal patterns of some parasitic diseases of medical and veterinary importance. At the same time, the incorporation of GIS in this field shows the scarcity of the data and images available, which sometime discourage researchers that still look at GIS as a system too difficult and unusable for medical study. Aware of this problem and supported by success of earlier MMDb's for Africa, Asia and South America, the authors' aim is to construct and offer an MMDb for Europe. The initial MMDb is composed with vector images covering an area situated from -11 degrees-70 degrees N to 58 degrees-30 degrees E. Specifically, data layers include: a) Global Moderate-Resolution Imaging Spectroradiometer (MODIS) Normalized Difference Vegetation Index (NDVI) 16 days at 250 m spatial resolution designed to provide consistent spatial and temporal comparisons of vegetation conditions, supplied in the MMDb as seasonal and annual composite images from 2000 to 2003, b) MODIS Land Surface Temperature (LST) calculated from daytime and nighttime observations at 8 day intervals at 1 km spatial resolution, supplied in the MMDb as seasonal and annual composites images for day (maximum) temperatures, night (minimum) temperatures from 2000 to 2003, c) GTOPO30 Digital Elevation Model (DEM) at 1 km spatial resolution, d) United States Geological Survey (USGS) Land use/land cover scheme, e) USGS actual and potential evapotranspiration supplied for all 12 months as a grid at 50 km spatial resolution, f) USGS precipitation showing the amount of rainfall for all 12 months supplied as a grid at 50 km spatial resolution, g) USGS shapefiles of administrative and political boundaries, cities, towns, villages, lakes, rivers, streams, road, railroads and more. The MMDb projection will be in geographic latitude-longitude, decimal degree format. This global format is most commonly used for public access map database resources and can be readily re-projected as needed for compatibility with various national mapping systems. There is no "required" software, and end users need only common commercial GIS software packages that have mutual import-export functions. Additionally, the MMDb is meant to be a dynamic resource that end users may improve and modify with other regional data.

The perception of needs of relatives of patients admitted to the Coronary Care Units (CCUs) and Intensive Care Units (ICUs) of the Trentino region was investigated by interviewing 50 relatives and 72 nurses. The questionnaire used for the survey included a list of 31 potential needs of a relative. Relatives and nurses were asked to select 10 needs and rank them according to the perceived priority; the nurses were also requested to rate the level of satisfaction related to each need, on a scale from 1 to 4. The data were collected over a period of 50 days, from the end of May to the beginning of July. The priority ranking of needs by relatives was as follows: being sure that the patient receives the best possible care; that the relative could see a doctor at least once a day; and being contacted and informed on every clinically relevant change in the health-illness status of the hospitalised relative. There was a good correspondence between relatives and nurses' perceptions while no significant differences in the perception of needs was observed between relatives interviewed in CCUs and ICUs, neither between nurses with different levels of expertise. Nurses perceive that the priority needs identified by the relatives are reasonably satisfied.