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Background Immunotherapeutic drugs, such as domperidone, have been shown to be promising treatments against canine leishmaniosis (CanL), but limited data are available. The aim of this pilot study (therapeutic, prospective and non-controlled) was to evaluate the effect of domperidone on serum antibody titers of Leishmania infantum, globulins, gamma globulins, acute-phase proteins (e.g. C-reactive protein [CRP]), big endothelin-1 (big ET-1), serum creatinine (SC) and proteinuria in dogs with leishmaniosis affected by chronic kidney disease (CKD). Methods Dogs were recruited if “exposed” to or “infected” with L. infantum and affected by CKD (IRIS stage 1 [proteinuric] or IRIS stage 2–3a [SC Results Of the 14 recruited dogs, nine showed a statistically significant reduction in SC (χ2 = 9.1, df = 3, P = 0.028), but not in the urine protein/creatinine ratio (χ2 = 6.43, df = 3, P = 0.092). All dogs showed a significant reduction in antibody titers for L. infantum (χ2 = 9.56, df = 2, P = 0.008), globulins (χ2 = 11.08, df = 3, P = 0.011) and gamma globulins (χ2 = 12.38, df = 3, P = 0.006) during the study period. There was also a statistically significant reduction in CRP (χ2 = 16.7, df = 3, P = 0.001), but not in big ET-1 (χ2 = 2.04, df = 3, P = 0.563). Conclusions This study provides preliminary results on the ability of domperidone to improve SC and reduce anti-L. infantum antibody titers, globulins, gamma globulins and CRP in dogs with leishmaniosis and CKD. Graphical abstract

Background C‐reactive protein (CRP) and paraoxonase 1 (PON1) might increase and decrease in canine leishmaniasis (CanL), , and both can rapidly normalize after therapy. Recently, supplementation of domperidone with conventional therapy , increasing the activity of cells involved in acute phase responses in vitro. This combined therapy has been recommended to treat mild forms of CanL; however, no studies have investigated the effects of domperidone supplementation on early CRP or PON1 changes in dogs with CanL. Objectives The aim of this study was to evaluate whether domperidone, added to conventional treatments, modifies CRP concentration and PON1 activity kinetics in CanL dogs responsive to conventional therapy. Methods Serum CRP concentrations and PON1 activities were measured in dogs with mild CanL before (t‐0) and 3 (t‐1), 7 (t‐2), 14 (t‐3), and 21 (t‐4) days after treatment with N‐methylglucamine antimoniate and allopurinol alone (n = 18) or combined with domperidone (n = 18). Results C‐reactive protein concentrations increased at t‐1 in the domperidone group, especially when the CRP concentration at t‐0 was normal. However, the concentrations normalized at t‐4 in 18/18 dogs compared with 14/18 dogs not receiving domperidone. The median PON1 activity decreased at t‐1 in the domperidone group, and this decrease was more significant in dogs with normal PON1 activity at t‐0. Conclusions Based on these results, transient increases in CRP concentrations or decreases in PON1 activities after domperidone administration should not be erroneously interpreted as signs of a worsening disease process.

Bats play a key role as reservoir hosts of many emerging viral diseases with zoonotic potential. However, little is known about the laboratory reference intervals (RIs) of bats, especially Egyptian fruit bats ( Rousettus aegyptiacus). The aim of this study was to obtain haematological, biochemical and electrophoretic RIs from captive fruit bats. Blood was collected from 21 R. aegyptiacus (11 females and 10 males). Complete blood cell count was performed using an impedance cell counter followed by the morphologic analysis of blood smears. Clinical biochemistry was performed with an automated spectrophotometer and agarose gel electrophoresis was carried out with an automated instrument. Reference intervals were determined using the Reference Value Advisor V2.1, following the American Society for Veterinary Clinical Pathology guidelines. Possible differences related to sex or sexual maturity were also investigated. The RIs for most of the analytes investigated were similar to those of other types of bats and other mammalian species. Haematology revealed mild polychromasia and slightly lower haematocrit, haemoglobin, leukocyte and lymphocyte counts compared to other bats. Glucose levels varied possibly due to stress, the anaesthetic protocol and fasting time. Creatine kinase was higher, while triglycerides were lower compared with domestic mammals and other bats. No sex- or age-related differences were found. Serum protein electrophoresis showed five fractions (albumin, α-, β1-, β2- and γ-globulins). The values recorded in this study could be helpful as a reference biological dataset to monitor the health status of wild and captive R. aegyptiacus and, possibly, of other Chiroptera.

Paraoxonase 1 (PON1) is an inflammation marker associated with lipid oxidation and is used as a diagnostic marker in people. There is no information about the suitable substrate and analytic performance in cats, or its biological behavior compared with other inflammation markers. Our aims were to validate a paraoxon-based method to measure PON1 activity in feline serum, to assess stability of PON1 under different storage conditions and the impact of interfering elements, to determine a reference interval (RI) for healthy cats, and to correlate PON1 activity with 2 major acute-phase proteins. Intra- and inter-assay precision, accuracy, and RI were assessed using fresh serum. The same specimens were stored at room temperature, refrigerated, or frozen, and retested at defined intervals. Hemolysis, lipemia, and icterus were simulated to study interferences. PON1 results were compared to serum amyloid A (SAA) and alpha-1-acid glycoprotein (AGP) results. Analytical validation yielded precise and accurate results. PON1 activity is stable for up to 24 h at room temperature and up to 48 h at 4°C. Freezing at −20°C results in an increase after 72 h, with return to baseline values after 1 wk, that again increases after 6 mo. Only hyperlipemia interfered with PON1 activity. The RI based on 71 healthy cats was 58–154 U/L. PON1 activity was negatively correlated with AGP, but not with SAA. Serum PON1 activity can be measured accurately in cats, and it acts as a negative acute-phase protein.

Objectives Feline infectious peritonitis (FIP) is an immune-mediated disease initiated by feline coronavirus (FCoV) infection. To date, the only proven route of transmission is the faecal–oral route, but a possible localisation of FCoV in the reproductive tract of tom cats is of concern, owing to the involvement of the male reproductive tract during FIP and to the presence of reproduction disorders in FCoV-endemic feline catteries. The aim of the study was to investigate the presence and localisation of FCoV in semen and/or in the reproductive tract of tom cats, and its possible association with seroconversion and viraemic phase. Methods Blood, serum, semen and/or testicle samples were obtained from 46 tom cats. Serology was performed on 38 serum samples, nested reverse transcriptase PCR (nRT-PCR) and reverse transcriptase quantitative PCR (RT-qPCR) were performed on 39 blood samples and on 17 semen samples, and histology, immunohistochemistry and nRT-PCR were performed on 39 testicles. Results Twenty-four of 38 serum samples were positive on serology. Semen samples were negative on RT-PCR and RT-qPCR for FCoV, while all blood samples were negative at both molecular methods, except for one sample positive at RT-qPCR with a very low viral load. All testicles were negative at immunohistochemistry, while six were positive at nRT-PCR for FCoV. Serology and blood PCR results suggest that the virus was present in the environment, stimulating transient seroconversion. FCoV seems not to localise in the semen of tom cats, making the venereal route as a way of transmission unlikely. Although viral RNA was found in some testicles, it could not be correlated with the viraemic phase. Conclusions and relevance In the light of these preliminary results, artificial insemination appears safer than natural mating as it eliminates the direct contact between animals, thus diminishing the probability of faecal–oral FCoV transmission.

SARS‐CoV‐2 positive or seropositive owned cats have been reported worldwide. The detection of seropositive stray cats in the proximity of farms of infected minks, coupled with the demonstration of cat‐to‐cat transmission in experimental settings, raise the question whether stray cats may have an epidemiological role in the COVID‐19 pandemic and may act as sentinel for the circulation of SARS‐CoV‐2. The aim of this study was to evaluate the presence of SARS‐CoV‐2 RNA and anti‐SARS‐CoV‐2 antibodies in free roaming cats belonging to colonies located in an area highly affected by the COVID‐19 pandemic and to correlate the results with the positivity rate in people sharing the same area. Interdigital, cutaneous, oropharyngeal, nasal and rectal swabs, as well as blood samples, were collected from 99 cats living in colonies and admitted to our hospital for neutering. This caseload corresponds to the 24.2% of the feline population living in the 25 sampled colonies and to the 5.6% of all the free‐roaming registered cats. The presence of SARS‐CoV‐2 RNA in swabs was assessed using real time RT‐PCR. Anti‐SARS‐CoV‐2 serum antibodies were assessed using commercially available ELISA kits and confirmed by serum virus neutralization. In people, the SARS‐CoV‐2 positivity rate ranged from 3.0% to 5.1% (mean rate: 4.1%) and the seropositive rate from 12.1% to 16.3% (mean rate: 14.2%). Most of the colonies were in urban areas and resident cats had frequent contacts with external cats or people. A COVID‐19 positive caretaker was found, whereas all the cats were negative for SARS‐CoV‐2 RNA and seronegative. Although the negative results cannot exclude previous infections followed by decrease of antibodies, this study suggests that colony cats do not have an important epidemiological role in SARS‐CoV‐2 transmission dynamics. Further studies on larger caseloads are warranted, also in the light of the emerging new viral variants, on a One Health perspective.

Reliable diagnostic and prognostic markers of sepsis are lacking, but essential in veterinary medicine. We aimed to assess the accuracy of C-Reactive Protein (CRP), protein carbonyls (PCO) and paraoxonase-1 (PON1) in differentiating dogs with sepsis from those with sterile inflammation and healthy ones, and predict the outcome in septic dogs. These analytes were retrospectively evaluated at admission in 92 dogs classified into healthy, septic and polytraumatized. Groups were compared using the Kruskal–Wallis test, followed by a Mann–Whitney U test to assess differences between survivors and non-survivors. Correlation between analytes was assessed using the Spearman’s test, and their discriminating power was assessed through a Receiver Operating Characteristic (ROC) curve. PON1 and CRP were, respectively, significantly lower and higher in dogs with sepsis compared with polytraumatized and clinically healthy dogs (p < 0.001 for both the analytes), and also in dogs with trauma compared with healthy dogs (p = 0.011 and p = 0.017, respectively). PCO were significantly increased in septic (p < 0.001) and polytraumatized (p < 0.005) as compared with healthy dogs. PON1 and CRP were, respectively, significantly lower and higher in dogs that died compared with survivors (p < 0.001 for both analytes). Ultimately, evaluation of CRP and PON1 at admission seems a reliable support to diagnose sepsis and predict outcomes.

Este artículo se encuentra disponible en la siguiente URL: https://onlinelibrary.wiley.com/doi/epdf/10.1111/jsap.13237 Este es el post-print del siguiente artículo: Roura, X., Cortadellas, O., Day, M. J., Benali, S. L., Canine Leishmaniosis Working Group, & Zatelli, A. (2021). Canine leishmaniosis and kidney disease: Q&A for an overall management in clinical practice. The Journal of Small Animal Practice, vol. 62, i. 1 (jan.), pp. E1?E19, que se ha publicado de forma definitiva en https://doi.org/10.1111/jsap.13237 This is the peer reviewed version of the following article: Roura, X., Cortadellas, O., Day, M. J., Benali, S. L., Canine Leishmaniosis Working Group, & Zatelli, A. (2021). Canine leishmaniosis and kidney disease: Q&A for an overall management in clinical practice. The Journal of Small Animal Practice, vol. 62, i. 1 (jan.), pp. E1?E19, which has been published in final form at https://doi.org/10.1111/jsap.13237 En este artículo de investigación también participan: Canine Leishmaniosis Working Group.

In people, serum Protein Carbonyls (PCOs) increase during oxidative stress (OS) due to oxidative damage to proteins. OS is often associated with inflammation and especially with sepsis, a condition hard to diagnose in veterinary medicine because reliable markers are lacking. The aim of this study was to assess whether PCOs in canine serum may be detected by antibody-based methods such as Western Blotting (WB), and to preliminarily investigate the possible utility of this marker in dogs with inflammation. A serum sample oxidized in vitro was used to set up the method; the coefficient of variation obtained by repeated analysis varied from 24 to 36%. In order to assess whether the technique may cover the range of PCOs concentration detectable in routine practice, PCOs were measured in 4 healthy dogs and in 15 with inflammatory diseases, in some cases potentially associated with sepsis, as suggested by the results of other inflammatory markers such as C-Reactive Protein (CRP) and the anti-oxidant enzyme Paraoxonase 1 (PON-1): the concentration of PCOs was low in dogs with normal PON-1 activity, moderately increased in the majority of dogs with low-normal PON-1 activity, and severely increased in dogs with very low PON-1 activity. In conclusion this study demonstrates that PCOs, may be detected in canine serum, using antibody-based techniques such as WB. The preliminary results in dogs with and without systemic inflammation encourage further studies on the possible role of PCOs as inflammatory markers.

SARS-CoV-2 emerged from animals and is now easily transmitted between people. Sporadic detection of natural cases in animals alongside successful experimental infections of pets, such as cats, ferrets and dogs, raises questions about the susceptibility of animals under natural conditions of pet ownership. Here, we report a large-scale study to assess SARS-CoV-2 infection in 919 companion animals living in northern Italy, sampled at a time of frequent human infection. No animals tested PCR positive. However, 3.3% of dogs and 5.8% of cats had measurable SARS-CoV-2 neutralizing antibody titers, with dogs from COVID-19 positive households being significantly more likely to test positive than those from COVID-19 negative households. Understanding risk factors associated with this and their potential to infect other species requires urgent investigation., SARS-CoV-2 can infect cats and dogs, but the extent to which pets are infected in households remains unclear. Here, Patterson et al. test 919 companion animals in northern Italy and find that some dogs and cats from COVID-19 positive households can test positive for COVID-19 neutralizing antibodies, with dogs significantly more likely to do so if they came from COVID-19 positive households.

Made available in DSpace on 2021-06-25T10:15:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-12-01 Background: Paraoxonase-1 (PON-1) is an antioxidant enzyme, whose activity decreases during the acute phase response in many species. Little is known about PON-1 and its role as a negative acute phase protein during septic inflammation in horses, but promising findings about its utility in diagnosing SIRS and predicting the outcome in diseased horses, were recently highlighted. The objective of the study was to investigate the behaviour of PON-1 in horses after experimentally induced endotoxemia. To this aim, PON-1 activity was measured on 66 plasma samples collected from six clinically healthy mares, previously included in another study, before and at multiple time points between 12 and 240 h after intravenous infusion of Escherichia coli O55:B5 lipopolysaccharide (LPS). Results: Compared with baseline values, a progressive transient decrease of PON-1 activity was observed starting from 24 h post-infusion, with lowest values observed between 3 to 7 days post-infusion, followed by a normalisation to pre-infusion levels the tenth day. Conclusions: The results of this study suggest that measurement and monitoring of PON-1 activity might be useful to evaluate progression and recovery from endotoxemia in horses. Further studies in horses with naturally occurring sepsis are warranted. Department of Veterinary Medicine Veterinary Teaching Hospital University of Milan Department of Veterinary Sciences Veterinary Teaching Hospital Univeristy of Pisa Department of Veterinary Clinical Science School of Veterynary Medicine and Animal Sicence Sao PAulo State University (Unesp) School of Veterinary Science Veterinary Teaching Hospital University of Sydney Department of Veterinary Clinical Science School of Veterynary Medicine and Animal Sicence Sao PAulo State University (Unesp)

Quantitative anti-Leishmania antibody titres are critical in the management of dogs with leishmaniosis, from diagnosis to treatment and follow-up, and there is a paucity of data relating changes in antibody titres to sand fly vector seasonality. This study aimed to evaluate seasonal variations in anti-Leishmania infantum antibody titres in dogs from a hyperendemic area for canine leishmaniosis (CanL). Leishmania infantum-seropositive and clinically healthy dogs (n=65) were sampled in June 2019 (sand fly season) and again in February-March 2020 (non-transmission season) to monitor clinical status and serological titres. There was a reduction in anti-L. infantum antibody titres during the non-transmission season in most dogs (n=36; 55.4%), and 44% of those dogs (n=16/36) became seronegative (i.e. below the cut-off value of 1:80). Given the relevance of serology to epidemiological, preventive and clinical studies related to CanL, seasonal variations in antibody titres are important in areas where phlebotomine vectors have seasonal patterns of activity. Sand fly seasonal period must be considered in the interpretation of annual anti-L. infantum antibody screening test results in asymptomatic dogs, to make clinical decisions about staging, treatment and prevention.

α1-Acid glycoprotein (AGP) is an acute-phase protein (APP) that modulates immune responses, probably – at least in humans – owing to the modification of its glycosylation pattern. On this perspective, feline AGP can be a useful comparative model, as it has different concentrations in cats susceptible or resistant to some disease. As a preliminary approach to the study of feline AGP (fAGP) we have purified this protein from feline serum by HPLC using human AGP (hAGP) as a model. Immunoblotting with a polyclonal antibody against fAGP and with a monoclonal antibody against hAGP was performed on serum from healthy cats, from cats exposed to feline coronavirus (FCoV) infection and from cats with purulent inflammations, such as feline infectious peritonitis (FIP), feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV). Immunohistochemistry on tissues from healthy cats and from cats with different diseases (FIP, FIV, FeLV, locally extensive inflammation) was also performed with the same antibodies. Both hAGP and fAGP have been purified to homogenity as determined by SDS-PAGE. fAGP did not react with the anti-hAGP antibody which, in contrast, detected in feline serum a low MW protein that we called fAGP-related protein (fAGPrP). This protein was underexpressed in cats with FeLV and FIP. Both fAGP and fAGPrP were immunohistochemically detected in plasma and hepatocytes with a stronger intensity in cats with FIP and some inflammatory conditions. Moreover, fAGPrP was detected in the cytoplasm of tissue cells, most likely identifiable with plasma cells. These cells were rarely detectable in cats with FIV and FeLV, and numerous in cats with FIP and with locally extensive inflammation. In conclusion, purified fAGP has physicochemical characteristics similar to those of hAGP, but does not cross-react with anti-hAGP antibodies. In contrast, the anti-hAGP detected an AGP-related protein whose blood concentration and tissue distribution was not related to that of fAGP. Moreover, both fAGP and fAGPrP were differently expressed in cats with pathologic conditions compared to controls. Further study of these proteins by analysing their structural characteristics is required.

Paraoxonase-1 (PON-1) is an anti-oxidant compound considered as negative acute phase protein in animals (Rossi et al., 2013) and people (Novak et al., 2010). The paraoxon-based method for measurement of PON-1 in equine serum has not yet been validated. The aim of this study is to validate a paraoxon-based method to measure PON-1 and to establish reference intervals (RIs) in healthy horses and foals. 120 horses (40 geldings, 40 stallions, 40 mares; median age: 11 years; 57 Warmbloods, 46 Trotters) and 55 foals (27 females, 28 males; median age: 47 days; 22 Warmbloods, 31 Trotters) considered healthy after physical examination and biochemistry were examined. Horses were grouped by breed: Thoroughbreds, Trotters, Warmbloods, Draft horses and Ponies. Serum PON-1 was measured with an automated spectrophotometer and an enzymatic method validated in other species (Giordano et al., 2013). After the analytical validation (precision, accuracy, interference studies), RIs were determined using the Reference Value Advisor software, according to ASCVP guidelines (Friedrichs et al., 2012). The possible gender-, age- and breed-related differences were statistically investigated. The paraoxon-based method was precise (CVs, International Journal of Health, Animal Science and Food Safety, V. 4, N. 1s (2017)

Background Increased serum activity of CK isoenzymes and macroenzymes, and in particular of the brain isoenzyme (CK-BB) has been reported in dogs with central nervous system (CNS) disorders. However, no studies on the possible differences in serum activities of CK iso- or macroenzymes (Macro-CK1 and Macro-CK2) in different neurologic diseases are available. Objective The aim of this study was to describe the electrophoretic distribution of CK iso- and macroenzymes in dogs with CNS disorders in order to assess whether this distribution depends on a specific neurologic disease. Methods This study was done on sera from 45 dogs with neurologic diseases (degenerative, n = 7; idiopathic epilepsy [IE], n = 14; inflammatory, n = 16; space occupying lesions [SOL], n = 8) and from 10 clinically healthy dogs. The separation of serum CK isoenzymes and macroenzymes was performed using an automated electrophoretic method already validated in dogs. Results Compared with healthy dogs, dogs with CNS disorders had significantly higher total CK and CK-BB activities, and a significantly lower Macro-CK2 activity (P < .001). Comparison of pathologic subgroups and healthy dogs revealed significant differences (P < .01) in dogs with IE and inflammatory disorders for total CK activity, in all the subgroups for CK-BB (P < .01), and in dogs with IE and SOL for Macro-CK2 (P < .01). Conclusions The results of this study suggest that CK-BB is released by neurons damaged by inflammatory or degenerative conditions or due to compressive effects of SOL. However, the neurologic diseases cannot be differentiated based on CK-BB or Macro-CK2 activities, unless further studies allow the definition of diagnostic thresholds.

Feline coronaviruses (FCoV) colonize the intestinal tract, however, due to not fully understood mutations, they can spread systemically and cause feline infectious peritonitis (FIP). Recent studies on human medicine report that gut microbiota is involved in the development of systemic disorders and could influence the immune response to viral diseases. The aim of this study was to provide preliminary data on the fecal microbiota composition in healthy cats compared to FCoV-infected cats, with and without FIP. Cats were equally grouped as healthy FCoV-negative, healthy FCoV-positive or FIP affected (total n = 15). Fecal sample were evaluated for the microbiota composition. A total of 3,231,916 sequences were analyzed. The samples' alpha-diversity curves did not reach a proper plateau and, for the beta-diversity, the samples seemed not to group perfectly by category, even if the healthy FCoV-positive group showed a hybrid microbial composition between FCoV-negative and FIP groups. Although there were no taxa significantly linked to the different conditions, some peculiar patterns were recognized: Firmicutes was always the most represented phylum, followed by Bacteroidetes and Actinobacteria. In FCoV-positive cats, the Firmicutes and Bacteroidetes were respectively over- and under-represented, compared to the other groups. Among FIP cats, three subjects shared a similar microbiome, one cat showed a different microbial profile and the other one had the lowest number of diverse phyla. Despite the limited number of animals, some differences in the fecal microbiome between the groups were observed, suggesting to further investigate the possible correlation between gut microbiota and FCoV infection in cats., Highlights • Mutated form of enteric feline coronaviruses (FCoV) may spread systemically inducing feline infectious peritonitis (FIP). • Gut microbiota is known to be influenced by viral diseases in people and in some animal species. • Fecal microbiota was evaluated in 15 cats divided in three groups as healthy, positive to FCoV and affected by FIP. • Some microbiota peculiarities have been observed associated with the presence of coronavirus and/or disease. • Feline gut microbiota composition may be associated with FCoV infection and thus need to be further investigated.

Feline infectious peritonitis (FIP) is characterised by the presence of systemic inflammation accompanied by oxidative stress. Paraoxonase-1 (PON-1) is a negative acute phase reactant produced by the liver. A paraoxon-based method has been validated to measure PON-1 activity in feline serum. The aim of this study was to investigate the usefulness of PON-1 activity as a biomarker to discriminate FIP from other diseases with similar clinical signs. Of 159 cats enrolled, 71 were healthy, 34 had FIP and 54 had another disease but presented with clinical signs that could be consistent with FIP. PON-1 activity was lower (P

Objectives The aim of this study was to evaluate whether the frequency of electrophoretic changes in serum of cats with feline infectious peritonitis (FIP) changed in recent years vs past years. Methods Agarose gel electrophoresis (AGE) and capillary zone electrophoresis (CZE) from cats with FIP and healthy cats recorded in the periods 2004–2009 and 2013–2014 were retrospectively analysed. Relative and absolute values of each electrophoretic fraction were recorded and the number of cats showing single or combined electrophoretic changes consistent with FIP (hypoalbuminaemia, inverted albumin to globulin [A:G] ratio, increased total protein, total globulin, alpha [α]2-globulin and gamma [γ]-globulin concentration) were counted. Additionally, a visual analysis of electrophoretograms was also performed. Results for the two time periods were statistically compared. Results The details of 91 AGE procedures (41 from cats with FIP and 50 from healthy cats) and 45 CZE procedures (26 from cats with FIP and 19 from healthy cats) were obtained from the database. No significant differences between the two time periods were found both in FIP and in healthy cats analysed with CZE and in healthy cats analysed with AGE. Compared with 2004–2009, cats with FIP sampled in 2013–2014 with AGE showed a significantly lower concentration of total protein, γ-globulins and total globulins, and a significantly higher A:G ratio and percentage of albumin and α2-globulins. Using both AGE and CZE, in recent years the proportion of cats with high α2-globulins without gammopathy and the proportion of cats with gammopathy alone decreased. With a visual approach, the number of patterns considered as dubious increased in the second period with AGE (non-statistically significant). Conclusions and relevance The frequency of electrophoretic abnormalities in cats with FIP decreased in recent years, independently of the technique employed. Although the mechanism responsible for this change was not investigated in this study, this altered frequency may decrease the diagnostic accuracy of serum protein electrophoresis for FIP.

There are several topical ectoparasiticides, drugs, supplements and vaccines that protect dogs against sand fly bites and/or reduce the risk of Leishmania infantum infection. This large variety of products and the absence of comparative studies mean that veterinarians are often faced with uncertainty. The aim of the study was to characterise the attitudes of veterinarians towards leishmaniosis prevention in client-owned dogs in Italy. A web-based questionnaire was prepared with 28 questions covering the working environment of veterinarians, advice given to owners regarding topical ectoparasiticides, advice on systemic prevention, and the use of diagnostic tests on which to base advice for prevention. The questionnaire was emailed to 9,426 veterinarians and answers were collected after 100 days. A total of 542 questionnaires were returned (response percentage 5.8 %): 54.8 % of the veterinarians considered their working area to be endemic, 29.3 % examined dogs from urban areas, and 42.3 % diagnosed ≥10 dogs/year with leishmaniosis. Those veterinarians who diagnosed ≥10 dogs/year with leishmaniosis were more likely to consider their working area to be endemic, whereas those who examined dogs mainly from urban areas were less likely to consider the area to be endemic. Veterinarians who considered the working area to be endemic were more prone to prescribe ectoparasiticides throughout the year, including collars and spot-on products and a combination of the two, vaccine alone or vaccine and domperidone combined, and used qualitative serological Leishmania tests. The attitude of veterinarians did not differ whether dogs were from urban or rural areas. In conclusion, veterinarians who consider their working area to be endemic or who frequently diagnose dogs with leishmaniosis tend to adopt more comprehensive strategies, and their attitudes are not influenced by the origin of the dogs: urban or rural. Overall, in Italy the attitudes of veterinarians in relation to canine leishmaniosis prevention appear to vary considerably.

Feline coronavirus (FCoV) is responsible, along with an inadequate immune response of the host, for Feline infectious peritonitis (FIP), one of the most frequent and deadly infectious feline disease worldwide. This study analyzed the genetic characteristics of the spike (S) gene of 33 FCoVs circulating in Northern Italy between 2011 and 2015 in cats with or without FIP. In order to reconstruct the most probable places of origin and dispersion of FCoV among Italian cats, a phylogeographic approach was performed based on 106 FCoV S gene partial sequences from cats, including the 33 novel Italian sequences and 73 retrieved from public databases. Only FCoV type I was found in the Italian cats. The estimated mean evolutionary rate of FCoV was 2.4 × 10-2 subs/site/year (95% HPD: 1.3-3.7 × 10-2), confirming the high genetic variability in the circulating strains. All the isolates clustered in a unique highly significant clade that likely originated from USA between the 1950s and the 1970s, confirming the first descriptions of the disease in American cats. Our results suggest that from USA the virus likely entered Germany and thereafter spread to other European countries. Phylogeography showed that sequences segregated mainly by geographical origin. In the 2010s Italian sequences clustered in different subclades, confirming that different strains cocirculate in Italy. Further studies on archival samples and other genetic regions of FCoV are suggested in order to confirm the present results and to reconstruct a more in-depth detailed virus dispersion pattern for the definition of possible control measures.

Highlights • Only 2/27 kidneys and 1/81 urine samples were FeMV PCR positive. • All cats with chronic kidney disease were FeMV PCR negative on urine. • Only 1/14 cats with proteinuria was FeMV positive on urine. • The 2 FeMV PCR positive kidneys were from cats affected by FIP and adenocarcinoma. • Italian FeMV strains form a sub-cluster with strains isolated from other countries., Feline morbillivirus (FeMV) is an emerging virus that was first described in Hong Kong in 2012. Several reports suggested the epidemiological association of FeMV infection with chronic kidney disease (CKD) in cats. The aim of this study was to investigate the presence and the genetic diversity of FeMV as well as the relationship between FeMV infection and CKD in cats from Northern Italy. Urine (n = 81) and kidney samples (n = 27) from 92 cats admitted to the Veterinary Teaching Hospital of the University of Milan between 2014 and 2017 were investigated for FeMV infection. FeMV RNA was detected in one urine sample (1.23%; 95% CI: 0.03–6.68%) and in two kidneys (7.40%; 95% CI: 0.91–24.28%). FeMV RNA was revealed only in urine or kidneys of cats without evidence of CKD. Phylogenetic analysis showed that the three strains clustered with FeMV strains retrieved from public database, forming a distinct sub-cluster of FeMV. The presence of distinct genotypes of FeMV found in this study is in accordance with previous studies demonstrating that FeMV strains are genetically diverse. A clear relationship between the presence of FeMV infection and CKD in the cats from Northern Italy was not observed, confirming recent reports that do not support the hypothesis that FeMV infection is associated with the development of CKD.

Objectives The increased demand for animal blood transfusions creates the need for an adequate number of donors. At the same time, a high level of blood safety must be guaranteed and different guidelines (GLs) deal with this topic. The aim of this study was to evaluate the appropriateness of different GLs in preventing transfusion-transmissible infections (TTI) in Italian feline blood donors. Methods Blood samples were collected from 31 cats enrolled as blood donors by the owners’ voluntary choice over a period of approximately 1 year. Possible risk factors for TTI were recorded. Based on Italian, European and American GLs, specific TTI, including haemoplasmas, feline leukaemia virus (FeLV), feline immunodeficiency virus (FIV), Anaplasma phagocytophilum, Ehrlichia species, Bartonella species, Babesia species, Theileria species, Cytauxzoon species, Leishmania donovani sensu lato and feline coronavirus (FCoV), were screened. Rapid antigen and serological tests and biomolecular investigations (PCR) were used. Several PCR protocols for haemoplasma and FeLV DNA were compared. Results The presence of at least one recognised risk factor for TTI was reported in all cats. Results for FIV and FeLV infections were negative using rapid tests, whereas five (16.1%) cats were positive for FCoV antibodies. Four (12.9%) cats were PCR positive for haemoplasma DNA and one (3.2%) for FeLV provirus, the latter being positive only using the most sensitive PCR protocol applied. Other TTI were not detected using PCR. Conclusions and relevance Blood safety increases by combining the recommendations of different GLs. To reduce the risk of TTI, sensitive tests are needed and the choice of the best protocol is a critical step in improving blood safety. The cost and time of the screening procedures may be reduced if appropriate tests are selected. To this end, the GLs should include appropriate recruitment protocols and questionnaire-based risk profiles to identify suitable donors.

Background The ΔWBC (the ratio between DIFF and BASO counts of the Sysmex XT‐2000iV), hereafter defined as ΔTNC (total nucleated cells), is high in effusions due to feline infectious peritonitis (FIP), as cells are entrapped in fibrin clots formed in the BASO reagent. Similar clots form in the Rivalta's test, a method with high diagnostic accuracy for FIP. Objectives The objective of this study was to determine the diagnostic accuracy for FIP and the optimal cutoff of ΔTNC. Methods After a retrospective search of our database, DIFF and BASO counts, and the ΔTNC from cats with and without FIP were compared to each other. Sensitivity, specificity, and positive and negative likelihood ratios (LR+, LR−) were calculated. A ROC curve was designed to determine the cutoff for best sensitivity and specificity. Results Effusions from 20 FIP and 31 non‐FIP cats were analyzed. The ΔTNC was higher (P 2.5 had 100% specificity. Conclusions The ΔTNC has a high diagnostic accuracy for FIP‐related effusions by providing an estimate of precipitable proteins, as the Rivalta's test, in addition to the cell count. As fibrin clots result in false lower BASO counts, the ΔTNC is preferable to the WBC count generated by the BASO channel alone in suspected FIP effusions.

Background Conventional differential leukocyte counts do not enumerate hyposegmented neutrophils (Hypo-PMN), ie, immature neutrophils that already lost the band morphology but are not yet completely segmented. They may be early indicators of acute inflammation. Objectives The purpose of the study was to evaluate the analytic variability of counts of segmented neutrophils (Seg-PMN), band neutrophils (Band), hyposegmented neutrophils (Hypo-PMN), non-Bands (Hypo-PMN + Seg-PMN), and Young-PMN (Bands + Hypo-PMN) assess if Hypo- or Young-PMN identify inflammation better than Bands. Methods Neutrophil subpopulations were counted by 2 observers on 2 sets of 100 cells in 267 blood smears from dogs with changes potentially consistent with inflammation to calculate the intra- and inter-observer variability. Results Median intra-observer CVs were

Highlights • Holy Birman cats (HB) seem to be predisposed to feline infectious peritonitis (FIP). • In this study, indicators of innate and specific immunity were assessed in HB cats. • CD4/CD8, IFN-γ, IL12/IL4 were higher and IL-4 was lower in HB than in other breeds. • The rate of shedders, but not of seropositivity, was lower in HB than in other cats. • Th1 immunity seems to be stronger in HB. This may reduce the rate of shedders., The aim of this study was to assess whether Holy Birman cats (HB) have a peculiar immune profile and a higher rate of infection by feline coronaviruses (FCoV). Leucocyte and lymphocyte subsets, antibody titers, α1-acid glycoprotein (AGP), globulin fractions, IL-4, IL-12 and IFN-γ in blood and fecal FCoV excretion were determined in HB (n = 75) and in cats from other breeds (n = 94). Significantly higher CD4/CD8 ratio, IFN-γ concentration and IL12/IL4 ratio and significantly lower IL-4 concentration and proportion of shedders were found in HB than in other breeds. No other differences were found. In conclusion, this study did not provide evidence of peculiar immune profiles in HB, except for a prevalent Th1 profile, that may explain why in our caseload the rate of shedders was lower in HB than in other breeds.

This study was designed to determine if the activity of paraoxonase (PON1), an antioxidant enzyme that works as a negative acute phase reactant, is a better predictor for the clinical recovery of leishmaniotic dogs receiving standard treatments compared with inflammatory markers such as C reactive protein (CRP) and electrophoretic fractions. For this purpose we tested 20 healthy dogs (controls) and 39 leishmaniotic dogs classified as sick (group A, n=23) or severely sick (group B, n=16) and tested at admission and after 3, 7, 14, 21, 28, 35 and 42 days. At admission, CRP and electrophoresis were altered in both groups, while PON1 activity was abnormal only in group B. There were no differences related to the outcome (mortality, complications or time of recovery). PON1 activity normalized in about 2 weeks in dogs that had abnormal values at admission and a final positive outcome; CRP normalized in 4-6 weeks and electrophoretic fractions were still altered after 6 weeks. The results show that, at admission, inflammatory markers did not predict the outcome of leishmaniasis. PON1 activity decreased only in some dogs with systemic inflammation but not in those with mild leishmaniasis: when decreased, PON1 normalized earlier than other markers in dogs that responded to treatment. This finding most likely depends on the rapid decrease in oxidative phenomena. PON1 activity should therefore be tested on admission: if low values are recorded, severe inflammation may be suspected and PON1 measurement may be repeated during treatment to early identify responsive dogs.

The serum concentration of high density lipoproteins (HDLs) is affected by oxidative phenomena that characterize inflammation. The aims of this study were to assess the concentration of HDLs in dogs with leishmaniasis, and to investigate the correlation between HDL, C-reactive protein (CRP), globulin fractions, and activity of the antioxidant enzyme paraoxonase (PON1). The serum concentration of HDLs was also measured during treatment to assess whether it may be a good marker to monitor response to therapy. HDLs were measured in serum from 10 leishmaniotic dogs (7 classified as sick and 3 as severely sick according to the staging system of the Canine Leishmaniasis Working Group), at admission and during the follow up (3, 7, 14, 21, and 28 days after the beginning of treatment with antimonials and allopurinol). Concentrations of total cholesterol, PON1, CRP and globulin fractions were also measured. At admission, the concentration of HDLs did not differ between sick and severely sick dogs. The correlation between HDL, PON1, CRP, and globulin fractions was assessed on the whole set of data. There was a negative correlation (P < .05, r = -0.42) between total cholesterol and c-globulins, and positive correlations between HDL and PON1 (P = .002; r = 0.55), and HDLs and A/G ratio (P = .019, r = 0.44). There was a negative correlation between HDLs and CRP (r = -0.37, P =,058). HDLs increased in sequential samples collected during the follow up after 14 and 28 days (P = .033 and P = .024). These results support an influence of c-globulins on lipid metabolism as reported in people, and suggest that inflammation is associated with a decreased HDLs concentration, likely due to the oxidative phenomena that decrease PON1 activity. Moreover, these results suggest that HDLs may be useful to assess decreased oxidation/inflammation after successful treatments. In conclusion, HDLs may serve as an additional inflammation and oxidation marker, including dogs with leishmaniasis.

High density lipoproteins (HDLs) are pivotal in innate immunity and decrease in serum during inflammation. Several studies have been done about lipoprotein changes in transition cows but little is known about their changes in newborn calves. The aim of this study is to provide information about HDLs in newborn calves, by defining the possible age-related changes in healthy calves compared with adults and by assessing the possible differences in calves with inflammation. Lipoprotein electrophoretic separation (reported as percentages) and colorimetric measurement of HDL (HDL-C) were performed on healthy cows and calves in order to identify possible differences in the lipoprotein profile due to the age. Then, age-matched calves with inflammatory conditions were also evaluated. Results showed that in calves HDL% and VLDL% were lower (mean values ± SD: 77.6% ± 8.6% and 2.6% ± 2.5%, respectively) and LDL% was higher (19.7% ± 7.4%) than in adults (89.0% ± 3.9%; 5.2 ± 2.1% and 5.8% ± 3.1%, respectively). Sick calves revealed a decrease of both HDL% (mean values ± SD: 61.0% ± 22.1%) and HDL-C (22.8 ± 11.6 mg/dL) and an increase of VLDL% (12.1% ± 13.1%) compared with controls (77.6% ± 8.6%; 41.5 ± 11.2 mg/dL and 2.6% ± 2.5%, respectively). Paraoxonase-1 activity, influenced by inflammation and oxidation, was measured, and it appeared correlated with HDL% and HDL-C in sick calves. In conclusion, this study revealed that HDLs concentration in healthy calves is lower than in adults, and further decreases in calves with inflammation, likely due to oxidation.

Retained placenta (RP) is often diagnosed in high-yielding dairy cows and can negatively affect reproductive performances. The objective of the present study was to investigate the hematological and biochemical profile of cows with RP before and immediately after parturition, with particular emphasis on neutrophil counts, since a previous study demonstrated the presence of peripheral neutropenia in dairy cows with RP sampled a few days after parturition. Results from 12 Holstein cows affected by RP and from 17 clinically healthy controls sampled one week pre-partum, within 12h after calving and between 48 and 72h after parturition were compared between groups and over time. Compared with controls, cows with RP had lower lymphocyte counts before parturition, lower leukocyte and neutrophil counts at parturition, lower monocyte counts at all times, and higher β-hydroxybutyrate before and after parturition. Erythroid and biochemical parameters were similar over time in both groups, whereas RP cows did not show the increase of neutrophil counts that occurs in controls at parturition. Hence, the finding of a lower neutrophil count in a routinely hemogram performed at parturition could be used as an alarm signal suggesting to monitor the affected animals. Moreover, although the underlying pathogenetic mechanism should be better investigated, the present study describes for the first time the association between altered blood leukocyte concentrations at parturition in RP compared to control cows.

Although several reviews on canine leishmaniasis have been published, none thoroughly described clinicopathologic abnormalities and their clinical usefulness. The aim of this review was to provide information concerning current diagnostic tests relevant for clinical pathologists and from a practical perspective. Specifically, in canine leishmaniasis, nonregenerative normocytic normochromic anemia, thrombocytopenia, or leukogram changes may be present. Clinical chemistry and urinalysis may indicate renal dysfunction (azotemia, decreased urine specific gravity, proteinuria) and an inflammatory/immune response (increased acute phase proteins [APP] or α2 - and/or γ-globulins). Although a potential gammopathy is usually polyclonal, it may also appear oligo- or monoclonal, especially in dogs coinfected by other vector-borne pathogens. When lesions are accessible to fine-needle aspiration (lymphoadenomegaly, nodular lesions, joint swelling), cytology is strongly advised, as the presence of Leishmania amastigotes in a pattern of pyogranulomatous inflammation or lymphoplasmacytic hyperplasia is diagnostic. If the cytologic pattern is inconclusive, the parasite should be identified by histology/immunohistochemistry or PCR on surgical biopsies. Alternatively, cytology and PCR may be performed on bone marrow samples where amastigotes, along with erythroid hypoplasia, myeloid hyperplasia, plasmacytosis, or secondary dysmyelopoiesis can be observed. Dogs with overt leishmaniasis generally have high antibody titers, while low titers predominate in immunologically resistant infected dogs or in exposed dogs with no parasite confirmation. Quantitative serology is recommended in clinically suspect dogs as high-titer antibodies titers may confirm the clinical diagnosis. In confirmed and treated dogs, renal function and inflammatory/immune response variables should be periodically monitored.

Background Hereditary bisalbuminemia is a relatively rare anomaly characterized by the occurrence of two albumin fractions on serum protein separation by electrophoresis. In human medicine, it is usually revealed by chance, is not been clearly associated with a specific disease and the causative genetic alteration is a point mutation of human serum albumin gene inherited in an autosomal codominant pattern. This type of alteration is well recognizable by capillary zone electrophoresis (CZE), whilst agarose gel electrophoresis (AGE) not always produces a clear separation of albumin fractions. The aims of this study is to report the presence of this abnormality in two separate groups of related bottlenose dolphins and to compare the results obtained with capillary zone and agarose gel electrophoresis. Results Serum samples from 40 bottlenose dolphins kept under human care were analyzed. In 9 samples a double albumin peak was evident in CZE electrophoresis while no double peak was noted in AGE profile. Since only an apparently wider albumin peaks were noted in some AGE electrophoretic profiles, the ratio between base and height (b/h) of the albumin peak was calculated and each point-value recorded in the whole set of data was used to calculate a receiver operating characteristic curve: when the b/h ratio of albumin peak was equal or higher than 0.25, the sensitivity and specificity of AGE to detect bisalbuminemic samples were 87 and 63 %, respectively. The bisalbuminemic dolphins belong to two distinct families: in the first family, all the siblings derived from the same normal sire were bisalbuminemic, whereas in the second family bisalbuminemia was present in a sire and in two out of three siblings. Conclusions We report for the first time the presence of hereditary bisalbuminemia in two groups of related bottlenose dolphins identified by means of CZE and we confirm that AGE could fail in the identification of this alteration.

Objective To assess whether alpha-1-acid glycoprotein (AGP) can be detected on the membrane of feline circulating leucocytes. Design The presence of AGP on circulating leucocytes was investigated in both clinically healthy cats and cats with different diseases. A group of feline coronavirus (FCoV)-positive cats, comprising cats with feline infectious peritonitis (FIP) and cats not affected by FIP but seropositive for FCoV, were included in this study because the serum concentration of AGP increases during FCoV infection. Procedure Flow cytometry (using an anti-feline AGP antibody), serum protein electrophoresis, routine haematology and measurement of the serum AGP concentration were performed using blood samples from 32 healthy cats (19 FCoV-seropositive), 13 cats with FIP and 12 with other diseases (6 FCoV-seropositive). The proportion of cats with AGP-positive leucocytes in the different groups (e.g. controls vs sick; FIP vs other diseases, etc.) or in cats with different intensities of inflammatory response was compared using a Chi-square test. Results AGP-positive leucocytes were found in 23% of cats. Compared with controls, the proportion of patients with positive granulocytes and monocytes was higher among sick cats (especially cats with diseases other than FIP) and cats with high serum AGP concentration, but not in cats with leucocytosis or that were FCoV-seropositive. Conclusion AGP-positive leucocytes can be found in feline blood, especially during inflammation. Conversely, no association between AGP-positive leucocytes and FIP was found. Further studies are needed to elucidate the mechanism responsible for this finding and its diagnostic role in cats with inflammation.

Objectives: Feline infectious peritonitis (FIP) can be difficult to diagnose. Histopathology is considered the gold standard test but immunohistochemistry (IHC) is mandatory to confirm/exclude the disease. This study aimed to assess the performances of tests carried out in vivo or at postmortem examination in challenging cases in which FIP was confirmed or excluded based on IHC or on adequate follow-up. Methods: Twelve cases (four without FIP, eight with FIP) were retrospectively studied. Clinical findings, serum protein electrophoresis (SPE), analysis of the effusions (AE), antifeline coronavirus serology, serum concentration of α1-acid glycoprotein (AGP) and histopathology were classified as consistent, doubtful or non-consistent with FIP. Sensitivity, specificity and concordance (κ) with the final diagnosis were calculated. Results: Concordance was absent for serology (κ=−0·08) and AE (κ=−0·52), poor for histopathology (κ=0·09), fair for SPE (κ=0·25) and perfect for AGP (κ=1·00). Sensitivity was high for AGP (100%) and low for AE (50%), SPE (37·5%) and histopathology (37·5%). Specificity was high for AGP or histopathology (100%) and low for SPE (50%) and AE (0%). Clinical Significance: IHC must always be performed to confirm FIP. If this is not possible, when histopathology is controversial, elevated AGP concentrations may support the diagnosis of FIP.

To assess the possible role of the “brain isoenzyme” of creatine kinase (CK-BB) as an early diagnostic tools for cerebral coenurosis we examined sera from 68 sheep (10–13 months old) from a flock on which cerebral coenurosis was endemical. After sampling, sheep were slaughtered, and neurological lesions were classified as purulent-necrotic migratory lesions or cystic lesions. The discriminating power of CK-BB to diagnose cerebral coenurosis was assessed using a receiver operating characteristic (ROC) curve. The prevalence of cerebral coenurosis was 39.7% (27/68). Migratory lesions were found in 23 sheep (85.1%), cystic lesions in 3 sheep (11.2%) and both lesions were found in 1 sheep (3.7%). The most abundant isoenzymes detected both in controls and in sheep with cerebral coenurosis were CK-MM, Macro-CK2 and CK-BB. No significant differences regarding absolute values of CK-BB were found suggesting that CK-BB is not released in serum during cerebral coenurosis. Conversely, CK-MM was significantly lower ( P P P

Background: Serum protein electrophoresis is widely used for diagnostic and research purposes. Cellulose acetate (CAE) and agarose gel (AGE) electrophoresis are the most frequently used methods, but capillary zone electrophoresis (CZE) is beginning to be used more in veterinary laboratories. However, reference intervals for CZE in animals and comparison studies with the other electrophoretic techniques are lacking, compromising the diagnostic utility of CZE. Objectives: The aims of this study were to compare results obtained using CAE, AGE, and CZE; to establish reference intervals for CZE in dogs and cats; and to assess the capacity of CZE to detect abnormalities identified by AGE. Methods: Serum samples from 204 dogs, including 104 healthy animals, and 62 cats, including 28 healthy animals, were analyzed using automated systems for CAE, AGE, and CZE. Descriptive statistics and Passing–Bablok and Bland–Altman tests were used to compare results. For each technique, reference intervals were calculated based on results from healthy animals. Concordance between CZE and AGE in detecting pathologic changes was assessed using Cohen's k coefficient. Results: For most protein fractions, values obtained by CAE, AGE, and CZE were significantly different from each other, and constant and proportional errors were often detected. Nevertheless, reference intervals obtained by the 3 techniques overlapped. Moreover, Cohen's k coefficient demonstrated that the capacity of CZE and AGE to detect pathologic changes was comparable. Conclusions: CZE performs comparably to AGE and CAE as long as CZE-specific reference intervals are used for interpretation and distinctive visual patterns for albumin, gaps between fractions, and subpeaks found on CZE tracings are recognized. In addition, CZE offers several technical advantages, such as ease of use and complete automation.

Background: Information about the electrophoretic distribution of CK-MM, CK-MB, and CK-BB, serum creatine kinase (CK) isoenzymes that are indicators of skeletal muscle, cardiac muscle, and brain lesions, respectively, and CK macroenzymes (macro-CK1 and macro-CK2) in dogs and cats with and without central neurologic disease is scant and equivocal. Objectives: The objectives of this study were to describe the electrophoretic distribution of CK isoenzymes and macroenzymes in healthy dogs and cats and to provide a preliminary assessment of the utility of CK enzymatic electrophoresis in dogs and cats with central neurologic disease. Methods: Electrophoretic separation of serum CK isoenzymes and macroenzymes was performed on freeze-thawed serum samples from 20 healthy dogs and 3 dogs with central neurologic disease and from 14 healthy cats and 6 cats with neurologic feline infectious peritonitis (FIP). Electrophoretic separation was also performed on supernatants of homogenized brain, skeletal muscle, and cardiac muscle from both species, to assess the tissue distribution of isoenyzmes in dogs and cats. Results: CK-MM was the predominant isoenzyme in the serum of healthy dogs and cats, followed by macro-CK2 and CK-BB in dogs and by both macroenzymes in cats. In dogs, CK-MB was essentially absent from both serum and homogenized hearts. CK-BB increased in dogs with neurologic disease. In cats, CK-BB was essentially absent from serum, but was present in brain homogenates. Two of 6 cats with FIP had increased macro-CK1 and increased CK-BB activity. Conclusions: This study identified the electophoretic distribution of CK isoenzymes and macroenzymes of dogs and cats and provided encouraging data about the possible use of CK-BB as a biomarker for canine neurologic disorders, but not for FIP.

rug treatment of leishmaniasis in dogs is a chal-lenge for veterinary practitioners. Because of its complex pathogenesis, leishmaniasis may manifest with various clinical signs, ranging from mild and nonspe-cific to those reflecting severe involvement of several organs. The immune response plays an important role in the development, outcome, and response to treat-ment of

Objective—To evaluate a urine dipstick test as a possible replacement for urine protein-tocreatinine (UPC) ratio for identifying proteinuria in dogs. Sample Population—507 urine samples from adult dogs. Procedures—Urine dipstick, UPC ratio, specific gravity (USG), and sediment testing were performed on 507 samples. With UPC ratio as the reference criterion, diagnostic accuracy of the urine dipstick test was calculated for the entire data set and for urine samples grouped by USG (≤ 1.012 or > 1.012; < 1.030 or ≥ 1.030). A UPC ratio < 0.2 was used to indicate absence of proteinuria. Results—The sensitivity of the urine dipstick test for detection of proteinuria was > 90% when 0 mg of protein/dL (a 0+ result) was used to indicate a negative test result, and the specificity ranged from 40% to 60%, depending on the USG. Sensitivity decreased to a range of 56% to 81% when 30 mg of protein/dL (a 1+ result) was used as the cutoff, depending on the USG, but the specificity increased to > 90%. The likelihood of correctly identifying nonproteinuric dogs was low when the USG was ≤ 1.012, particularly when samples with a 1+ result were considered negative. Conclusions and Clinical Relevance—For dogs with a dipstick-test result of 1+ and USG ≤ 1.012, proteinuria should be assessed by use of the UPC ratio; dogs with a USG value > 1.012 are likely nonproteinuric. When used together, the urine dipstick test and USG measurement were reliable as a rapid alternative to UPC ratio determination in dogs in this study.

The sialylation pattern of serum α1-acid glycoprotein (AGP) in non-symptomatic cats infected by feline coronavirus (FCoV) and its possible relationship with the amount of FCoVs shed in faeces were investigated. Blood from three specific pathogen-free cats (group A) and from 10 non-symptomatic FCoV-positive cats from catteries with low (group B, three cats) or high (group C, seven cats) levels of faecal shedding were collected monthly. AGP was purified from serum and Western blotting followed by lectin-staining of α(2,3)-linked and α(2,6)-linked sialic acid. Faecal shedding was quantified in group C by quantitative polymerase chain reaction. Variations of AGP sialylation were recorded only in cats from group C, on which viral shedding peaked before the occurrence of feline infectious peritonitis (FIP) in the cattery, and decreased 1 month later, when serum AGP had an increase of α(2,3)-linked sialic acid. These results suggest that hypersialylation of AGP may be involved in host–virus interactions.

This report describes a case of feline infectious peritonitis (FIP) in a castrated cat which first presented with the unusual sign of priapism. Laboratory examinations showed increased serum protein content and decreased albumin/globulin ratio. Serum electrophoresis revealed increased α2- and γ-globulin content. One month after the first examination, the cat died. At necropsy, histopathological evaluation of organs showed inflammatory granulomatous lesions compatible with non-effusive FIP and coronavirus-specific polymerase chain reaction confirmed the diagnosis. FIP antigen was demonstrated immunohistochemically in penile tissue.

Previous studies have demonstrated that the concentration of α1-acid glycoprotein (AGP) transiently increases in asymptomatic cats infected with feline coronavirus (FCoV). In order to establish whether these fluctuations depend on the FCoV status, the serum concentration of AGP and anti-FCoV antibody titres and/or faecal shedding of FCoVs in clinically healthy cats from catteries with different levels of prevalence of FCoV infection were monitored over time. Serum AGP concentrations fluctuated over time in clinically healthy cats from the cattery with the highest prevalence of feline infectious peritonitis (FIP) and significantly increased just before an outbreak of FIP. Further studies are required to clarify whether the observed increase of AGP concentration is a consequence of the increased viral burden or a protective response against mutated viral strains. Nevertheless, the results of the present study suggest that AGP might be useful in monitoring FCoV–host interactions in FCoV-endemic catteries.

Administration of recombinant feline interferon-omega (rFeIFN) has been proposed for the prophylaxis of canine and feline parvovirosis. In the present study, the influence of the administration of rFeIFN on blood markers of inflammation (alpha-globulins, alpha(1)-acid glycoprotein) and immune system activation (gamma-globulins, IgG, IgM, specific anti-feline parvovirus IgG or IgM) was evaluated in a cattery developing an outbreak of feline panleukopenia due to feline parvovirus (FPV) infection few days after initial administration of rFeIFN. Kittens (n=23) were injected with rFeIFN (1MU/kg subcutaneously, once a day for 3 days) and their blood parameters were compared with those of 17 untreated cats. Cats that survived the outbreak were vaccinated and re-sampled 1 month after the last rFeIFN administration. Time of emergence of clinical signs and survival rate were not significantly different between the two groups. Controls and treated cats surviving the infection had high levels of gamma-globulins, total- and anti-FPV specific IgGs, likely due to passive transfer of maternal immunity. Compared to controls, treated kittens had lower levels of alpha(1)-globulins and higher mean values of gamma-globulins and immunoglobulins. Data from samples collected after vaccination revealed a higher level of gamma-globulins, total- and anti-FPV specific IgGs in treated kittens, compared with controls, suggesting that rFeIFN stimulates antibody production. Based on this results, rFeIFN should be administered to the queen, to increase passive maternal immunity, or to kittens before introduction in a potentially contaminated environment.

This review presents the consensus opinion of the Canine Leishmaniasis Working Group on the prognosis and monitoring of leishmaniasis in dogs. While the prognosis for both exposed and infected dogs is considered to be favourable, this changes if infection progresses to overt disease. For clinically affected animals undergoing treatment, the prognosis is dictated by the severity of the signs (and in particular the severity of renal dysfunction) when therapy is initiated; assessing the degree of proteinuria is useful in this context. Approximately 75% of dogs without evidence of renal involvement live for >4years if adequately treated. Monitoring the response to treatment includes ongoing clinical and clinicopathological assessment, as well as quantifying serological responses and the parasite load in the patient.

In this study, the cytokine profiles of clinically healthy cats naturally infected with feline coronavirus (FCoV), of cats with feline infectious peritonitis (FIP) and of specific pathogen-free (SPF) cats were investigated in whole blood using a traditional reverse-transcriptase polymerase chain reaction (RT-PCR) assay and a semi-quantitative method of analysis based on computerised quantification of positive bands. The low inter-assay coefficient of variation recorded demonstrated that this method is highly repeatable. Compared with SPF cats, cytokine production was upregulated in most of the samples from FCoV-positive non-symptomatic cats. The appearance of a case of FIP in the cattery was associated with an increased expression of cytokines, in particular there was an increased production of IL-1β and IFN-γ, suggesting that these cytokines might protect infected cats from the disease. This hypothesis was also supported by the low levels of IFN-γ recorded in blood from cats with FIP.

Serum concentrations of acute-phase proteins (APPs): haptoglobin (Hp), ceruloplasmin (Cp), serum amyloid A (SAA), and C-reactive protein (CRP) were determined in healthy dogs (n = 15) and dogs with different diseases grouped as acute inflammation (I, n = 12), hematologic neoplasias (HT, including leukemia and lymphoma, n = 16), nonhematologic neoplasias (NHT, including epithelial, mesenchymal, and mixed, n = 20), and autoimmune hemolytic anemia (AIHA, n = 8). SAA and CRP were analyzed using commercially available enzyme-linked immunosorbent assay (ELISA) kits, and Hp and Cp were measured using colorimetric methods, all previously validated for use in dogs. Increased concentrations of all APPs were observed in all groups of diseased dogs, but statistical significance only was observed with Hp (I, P < .001; HT, P < .05), Cp (I, P < .05; AIHA, P < .01), and CRP (I, P < .001; HT, P < .001; AIHA, CRP P < .05). High variability in individual APPs within each group of diseases was found with no significant differences between leukemia and lymphoma as well as among different types of neoplasia. The AIHA group had smaller increases in Hp, SAA, and CRP but higher concentrations of Cp. When follow-up of individual cases was possible, a decrease in APPs generally was found in cases with favorable outcome. The results of this study suggest that neoplasia and hematologic diseases such as AIHA should be considered as possible causes of mild increases in APPs in dogs. Measurement of APPs may be helpful to assess clinical evolution and monitor treatment of these processes.

α1-Acid glycoprotein (AGP) is considered one of the major acute phase proteins in cats. In humans, AGP is a heavily glycosylated protein that undergoes several modifications of its glycan moiety during acute and chronic inflammatory pathologies. In this paper we present the feline AGPs (fAGP) glycan moiety modifications in the course of two prevalent feline diseases, the FIV (feline immunodeficiency virus) dependent feline acquired viral immunodeficiency and the feline leukemia virus (FeLV) associated lymphoma. The glycan moiety of fAGP was investigated by means of the binding of its oligosaccharides residues with specific lectins. Four lectins were used: Sambucus nigra agglutinin I and Maackia amurensis agglutinin lectins were used to detect sialic acid residues, Aleuria aurantia lectin was used to detect l -fucose residues and Concanavalin A was used to evaluate the degree of branching. It was found that fAGP undergoes several post-translational modifications of its glycan pattern: in particular the degree of sialylation is increased in FeLV-positive cats diagnosed with lymphoma, while FeLV-positive that did not presented any specific clinical signs cats do not present any increase of expression of sialic acid on the surface. Furthermore, FIV induced a modification of the glycan moiety of fAGP, which however varied widely among individuals. In order to determine the number and the position of oligosaccharide chains, the cDNA sequence of fAGP was also determined. The translation of the mature fAGP coding sequence gave rise to a sequence of 183 residues, with five potential N-glycosylation sites, but also with seven potential phosphorylation sites.

The immune system of vertebrates detects bacterial DNA as a "danger signal" based on the presence of unmethylated CpG motifs. We examined whether oligodeoxynucleotides (ODNs) with CpG motifs (CpG-ODNs) also induce mobilization of hematopoietic progenitor cells (HPCs). Mice challenged with CpG-ODNs showed an increase in peripheral blood colony-forming units (CFU) with a peak at day 4 after treatment, associated with an increase, starting 30 min after CpG treatment, in serum levels of mouse keratinocyte-derived chemokine (mKC), a functional homolog of human interleukin (IL) 8; production of granulocyte-colony-stimulating factor (CSF) was also detected. Mobilization and mKC induction were sequence-specific and dose-dependent occurring even with low doses of CpG-ODNs. Interestingly, intestinal cells were involved in mKC production. HPC mobilization by CpG-ODNs was dependent on peripheral blood mononuclear cells since mobilization was reduced in neutrophil-depleted mice. Moreover, CpG-ODN treatment significantly increased G-CSF mobilizing capacity. Finally, pretreatment with an anti-mKC neutralizing antibody significantly reduced CpG-induced mobilization, further supporting a role for mKC. Thus, bacterial DNA is a "danger signal" not only for immune cells but also for hematopoietic cells, communicating the need for increased hematopoiesis during infections and for the renewal of the immune system. The HPC mobilization activity of CpG-ODNs will need to be considered in the design of treatment regimens for cancer clinical trials using CpG-ODNs in association with chemotherapy.

Feline infectious peritonitis (FIP) is an immune-mediated disease of domestic and exotic felides infected with feline coronavirus. FIP is characterized by the overexpression of an acute phase protein, the alpha1-acid glycoprotein (AGP). In humans, AGP is a heavily glycosylated protein that undergoes several modifications of its glycan moiety during acute and chronic inflammatory pathologies. We studied the changes in AGP glycosylation in the course of FIP. Specifically, we focussed our attention on the degree of sialylation, fucosylation and branching. This study presents a purification method for feline AGP (fAGP) from serum, using an ion exchange chromatography strategy. The glycosylation pattern was analyzed in detail by means of interaction of purified fAGP with specific lectins. In particular, Sambucus nigra agglutinin I and Maackia amurensis agglutinin lectins were used to detect sialic acid residues, Aleuria aurantia lectin was used to detect L-fucose residues and Concanavalin A was used to evaluate the branching degree. By this method we showed that fAGP did not present any L-fucose residues on its surface, and that its branching degree was very low, both in normal and in pathological conditions. In contrast, during FIP disease, fAGP underwent several modifications in the sialic acid content, including decreased expression of both alpha(2-6)-linked and alpha(2-3)-linked sialic acid (76 and 44%, respectively when compared to non-pathological feline AGP).