Publications by authors named "Alessia Giordano"

59 Publications

Do Dogs and Cats Passively Carry SARS-CoV-2 on Hair and Pads?

Viruses 2021 07 13;13(7). Epub 2021 Jul 13.

Department of Veterinary Medicine, University of Milan, 26900 Lodi, Italy.

The epidemiological role of domestic animals in the spread and transmission of SARS-CoV-2 to humans has been investigated in recent reports, but some aspects need to be further clarified. To date, only in rare cases have dogs and cats living with COVID-19 patients been found to harbour SARS-CoV-2, with no evidence of pet-to-human transmission. The aim of the present study was to verify whether dogs and cats act as passive mechanical carriers of SARS-CoV-2 when they live in close contact with COVID-19 patients. Cutaneous and interdigital swabs collected from 48 dogs and 15 cats owned by COVID-19 patients were tested for SARS-CoV-2 by qRT-PCR. The time elapsed between owner swab positivity and sample collection from pets ranged from 1 to 72 days, with a median time of 23 days for dogs and 39 days for cats. All samples tested negative, suggesting that pets do not passively carry SARS-CoV-2 on their hair and pads, and thus they likely do not play an important role in the virus transmission to humans. This data may contribute to confirming that the direct contact with the hair and pads of pets does not represent a route for the transmission of SARS-CoV-2.
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http://dx.doi.org/10.3390/v13071357DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8310179PMC
July 2021

Maternal and neonatal canine cortisol measurement in multiple matrices during the perinatal period: A pilot study.

PLoS One 2021 22;16(7):e0254842. Epub 2021 Jul 22.

Department of Veterinary Medicine, Università degli Studi di Milano, Milan, Italy.

Stress exposure during perinatal period may lead to maternal cortisol increase that negatively affects the offspring development. In recent years, the interest on non-invasive sampling methods to measure cortisol as a marker of stress is increasing in both humans and animals. Indeed, discomfort due to blood collection may compromise the diagnostic outcome, mainly in uncooperative patients. So far, some alternative matrices but not milk have been explored in adult dogs, while no data are available on the neonate and paediatric live pups. This study aimed to measure cortisol concentration in different biological substrates in both dams (blood, saliva, hair and milk) and pups (saliva and hair) at established times from proestrus up to two months after parturition. For this purpose, five female German shepherd bitches and their 22 pups were enrolled. Cortisol concentration was assessed using the enzyme immunoassay kit (Salivary Cortisol ELISA kit, Salimetrics) after matrices appropriate preparation if required. Cortisol was measurable in all the substrates, except some milk samples below the detection limit. Maternal cortisol concentrations differed among the matrices (P <0.0001) with the highest values recorded in plasma (median 0.596 μg/dL) compared to saliva (median 0.159 μg/dL), hair (median 0.083 μg/dL) and milk (median 0.045 μg/dL). Cortisol in dams did not vary within the same matrix over time. In pups, salivary (median 0.295 μg/dL) cortisol was always higher than hair (median 0.049 μg/dL; P <0.0001). At birth (P = 0.01) and two months later (P = 0.05), neonatal salivary cortisol was higher compared to other samplings. The present study demonstrates the suitability of these innovative substrates for cortisol measurement, suggesting them as potential diagnostic support in canine neonatology and welfare.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0254842PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8297866PMC
July 2021

State of the art of F-FDG PET/CT application in inflammation and infection: a guide for image acquisition and interpretation.

Clin Transl Imaging 2021 Jul 10:1-41. Epub 2021 Jul 10.

Regional Center of Nuclear Medicine, Department of Translational Research and Advanced Technologies in Medicine, University of Pisa, Pisa, Italy.

Aim: The diagnosis, severity and extent of a sterile inflammation or a septic infection could be challenging since there is not one single test able to achieve an accurate diagnosis. The clinical use of 18F-fluorodeoxyglucose ([F]FDG) positron emission tomography/computed tomography (PET/CT) imaging in the assessment of inflammation and infection is increasing worldwide. The purpose of this paper is to achieve an Italian consensus document on [F]FDG PET/CT or PET/MRI in inflammatory and infectious diseases, such as osteomyelitis (OM), prosthetic joint infections (PJI), infective endocarditis (IE), prosthetic valve endocarditis (PVE), cardiac implantable electronic device infections (CIEDI), systemic and cardiac sarcoidosis (SS/CS), diabetic foot (DF), fungal infections (FI), tuberculosis (TBC), fever and inflammation of unknown origin (FUO/IUO), pediatric infections (PI), inflammatory bowel diseases (IBD), spine infections (SI), vascular graft infections (VGI), large vessel vasculitis (LVV), retroperitoneal fibrosis (RF) and COVID-19 infections.

Methods: In September 2020, the inflammatory and infectious diseases focus group (IIFG) of the Italian Association of Nuclear Medicine (AIMN) proposed to realize a procedural paper about the clinical applications of [F]FDG PET/CT or PET/MRI in inflammatory and infectious diseases. The project was carried out thanks to the collaboration of 13 Italian nuclear medicine centers, with a consolidate experience in this field. With the endorsement of AIMN, IIFG contacted each center, and the pediatric diseases focus group (PDFC). IIFG provided for each team involved, a draft with essential information regarding the execution of [F]FDG PET/CT or PET/MRI scan (i.e., indications, patient preparation, standard or specific acquisition modalities, interpretation criteria, reporting methods, pitfalls and artifacts), by limiting the literature research to the last 20 years. Moreover, some clinical cases were required from each center, to underline the teaching points. Time for the collection of each report was from October to December 2020.

Results: Overall, we summarized 291 scientific papers and guidelines published between 1998 and 2021. Papers were divided in several sub-topics and summarized in the following paragraphs: clinical indications, image interpretation criteria, future perspectivess and new trends (for each single disease), while patient preparation, image acquisition, possible pitfalls and reporting modalities were described afterwards. Moreover, a specific section was dedicated to pediatric and PET/MRI indications. A collection of images was described for each indication.

Conclusions: Currently, [F]FDG PET/CT in oncology is globally accepted and standardized in main diagnostic algorithms for neoplasms. In recent years, the ever-closer collaboration among different European associations has tried to overcome the absence of a standardization also in the field of inflammation and infections. The collaboration of several nuclear medicine centers with a long experience in this field, as well as among different AIMN focus groups represents a further attempt in this direction. We hope that this document will be the basis for a "common nuclear physicians' language" throughout all the country.

Supplementary Information: The online version contains supplementary material available at 10.1007/s40336-021-00445-w.
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http://dx.doi.org/10.1007/s40336-021-00445-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8271312PMC
July 2021

Absence of SARS-CoV-2 RNA and anti-SARS-CoV-2 antibodies in stray cats.

Transbound Emerg Dis 2021 Jun 25. Epub 2021 Jun 25.

Department of Veterinary Medicine, University of Milan, Italy.

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.
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http://dx.doi.org/10.1111/tbed.14200DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8446966PMC
June 2021

The Long Pentraxin PTX3 Controls Severe Infection.

Front Immunol 2021 20;12:666198. Epub 2021 May 20.

Department of Inflammation and Immunology, IRCCS Humanitas Research Hospital, Rozzano, Italy.

is a common pathogen in human sepsis. The emergence of multidrug-resistant strains represents a major clinical challenge in nosocomial and community acquired infections. The long pentraxin PTX3, a key component of humoral innate immunity, is involved in resistance to selected pathogens by promoting opsonophagocytosis. We investigated the relevance of PTX3 in innate immunity against infections using mice and mouse models of severe infections. Local and systemic PTX3 expression was induced following pulmonary infection, in association with the up-regulation of TNF-α and IL-1β. PTX3 deficiency in mice was associated with higher bacterial burden and mortality, release of pro-inflammatory cytokines as well as IL-10 in the lung and systemically. The analysis of the mechanisms responsible of PTX3-dependent control of infection revealed that PTX3 did not interact with , or promote opsonophagocytosis. The comparison of susceptibility of wild-type, and / mice to the infection showed that PTX3 acted in a complement-independent manner. Lung histopathological analysis showed more severe lesions in mice with fibrinosuppurative, necrotizing and haemorrhagic bronchopneumonia, associated with increased fibrin deposition in the lung and circulating fibrinogen consumption. These findings indicate that PTX3 contributes to the control of infection by modulating inflammatory responses and tissue damage. Thus, this study emphasizes the relevance of the role of PTX3 as regulator of inflammation and orchestrator of tissue repair in innate responses to infections.
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http://dx.doi.org/10.3389/fimmu.2021.666198DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8173212PMC
May 2021

Exploring alterations in hematological and biochemical parameters, enzyme activities and serum cortisol in Besnoitia besnoiti naturally infected dairy cattle.

Parasit Vectors 2021 Mar 15;14(1):154. Epub 2021 Mar 15.

Department of Veterinary Medicine, Università degli Studi di Milano, Via dell'Università 6, 26900, Lodi, Italy.

Background: Besnoitia besnoiti is an Apicomplexan protozoa causative of bovine besnoitiosis, a chronic and debilitating disease of cattle, with a variety of pathological findings that could alter some laboratory parameters. A study was conducted in a bovine besnoitiosis endemically infected dairy herd located in Italy characterized by high intra-herd seroprevalence and cattle with clinical signs of the disease. In the study, alterations in laboratory parameters, i.e. hematological and biochemical parameters, enzyme activities and serum cortisol levels, in Besnoitia besnoiti naturally infected cows were investigated in depth.

Methods: Laboratory parameters in 107 cows, of which 61 were seronegative and 46 were seropositive to B. besnoiti, including 27 with clinical signs of bovine besnoitiosis, were compared. Generalized linear models were used to evaluate the effect of Besnoitia infection on the considered laboratory parameters.

Results: Hematological analyses revealed that B. besnoiti infection determined a significant alteration to the leukocyte differential, with a higher percentage of granulocytes and a lower percentage of lymphocytes in seropositive and clinically affected animals (Mann-Whitney U-test, P = 0.022); erythrocyte and platelet counts did not show any difference between the considered groups of cows. Biochemistry tests evidenced that the parasite infection influenced serum protein values in seropositive cows and glutamate dehydrogenase values in clinically affected animals. No or only slight differences were revealed for all of the other biochemical and enzyme activity parameters in B. besnoiti-infected animals. In addition, despite the lack of statistical significance, seropositive and clinically affected cows evidenced higher concentrations of serum cortisol values compared to seronegative animals.

Conclusions: Although physiological, pathological and farm-related factors could have influenced the results in investigated animals, further studies involving more animals from different farms would be advisable to infer the role of B. besnoiti on these alterations, since laboratory parameters could help veterinarians in the diagnosis of bovine besnoitiosis in cattle.
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http://dx.doi.org/10.1186/s13071-021-04626-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7962361PMC
March 2021

Human-to-Cat SARS-CoV-2 Transmission: Case Report and Full-Genome Sequencing from an Infected Pet and Its Owner in Northern Italy.

Pathogens 2021 Feb 23;10(2). Epub 2021 Feb 23.

Luigi Sacco Department of Biomedical and Clinical Sciences, Università Statale di Milano, 20157 Milan, Italy.

There have been previous reports of the human-to-cat transmission of SARS-CoV-2, but there are only a few molecular studies that have compared the whole genome of the virus in cats and their owners. We here describe a case of domestic SARS-CoV-2 transmission from a healthcare worker to his cat for which nasopharyngeal swabs of both the cat and its owner were used for full-genome analysis. The results indicate that quarantine measures should be extended to pets living in SARS-CoV-2-infected households.
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http://dx.doi.org/10.3390/pathogens10020252DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7926546PMC
February 2021

Transplantation of autologous extracellular vesicles for cancer-specific targeting.

Theranostics 2021 1;11(5):2034-2047. Epub 2021 Jan 1.

Department of Health Sciences, University of Milan, Milan, Italy.

Nano- and microsized extracellular vesicles (EVs) are naturally occurring cargo-bearing packages of regulatory macromolecules, and recent studies are increasingly showing that EVs are responsible for physiological intercellular communication. Nanoparticles encapsulating anti-tumor theranostics represent an attractive "exosome-interfering" strategy for cancer therapy. : Herein, by labeling plasma-derived EVs with indocyanine green (ICG) and following their biodistribution by and imaging, we demonstrate the existence of nanoparticles with a highly selective cancer tropism in the blood of colorectal cancer (CRC) patients but not in that of healthy volunteers. : In CRC patient-derived xenograft (PDX) mouse models, we show that transplanted EVs recognize tumors from the cognate nanoparticle-generating individual, suggesting the theranostic potential of autologous EVs encapsulating tumor-interfering molecules. In large canine breeds bearing spontaneous malignant skin and breast tumors, the same autologous EV transplantation protocol shows comparable safety and efficacy profiles. : Our data show the existence of an untapped resource of intercellular communication present in the blood of cancer patients, which represents an efficient and highly biocompatible way to deliver molecules directly to the tumor with great precision. The novel EV-interfering approach proposed by our study may become a new research direction in the complex interplay of modern personalized cancer therapy.
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http://dx.doi.org/10.7150/thno.51344DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7797692PMC
August 2021

Prognostic impact of clinical, haematological, and histopathological variables in 102 canine cutaneous perivascular wall tumours.

Vet Comp Oncol 2021 Jun 8;19(2):275-283. Epub 2021 Jan 8.

Dipartimento di Medicina Veterinaria, Università degli Studi di Milano, Milan, Italy.

Identification of prognostic factors for perivascular wall tumours (PWTs) is desirable to accurately predict prognosis and guide treatment. 100 and two dogs with surgically excised PWTs without distant metastasis were retrospectively enrolled in this multi-institutional study, and the impact of pre-treatment leukocyte parameters, clinical and histopathological variables on local recurrence (LR) and overall-survival time (OST) were evaluated. Increasing values of white blood cell count (WBCC), neutrophil count (NC) and neutrophil-to-lymphocyte ratio (NLR) were significantly correlated with the hazard of LR in univariate analysis. WBCC and NC remained prognostic when adjusted for margins, grade, tumour size, location and skin ulceration, but lost their significance when adjusted for mitotic index and necrosis, whilst NLR remained prognostic only when close margins where categorised as infiltrated. Castrated males had a higher hazard of LR than intact males in univariate analysis, but significance was lost in multivariate models. Ulcerated PWTs and those located on the distal extremities had a higher hazard of LR both in univariate and multivariate analysis. Histological grade, necrosis, mitotic count, and infiltrated margins were all associated with LR both in univariate and multivariate analysis. Boxer breed, older age, ulceration, grade III, necrosis >50% and higher mitotic count were correlated with shorter OST, although breed and age lost their significance in multivariate analysis. Prognostication of surgically excised PWTs should be based on both clinical and histopathological variables. If validated in further studies, leukocyte counts and NLR may aid the clinician in identifying dogs at higher risk of LR before treatment.
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http://dx.doi.org/10.1111/vco.12673DOI Listing
June 2021

Analytical and Clinical Validation of a New Immunoenzymatic Method for the Measurement of Canine Parathyroid Hormone.

Animals (Basel) 2020 Dec 17;10(12). Epub 2020 Dec 17.

Department of Veterinary Medicine, University of Milan, Via dell'Università 6, 26900 Lodi, Italy.

Renal hyperparathyroidism (RHPT) is one of the main complications in dogs affected with Chronic Kidney Disease (CKD). The measurement of serum parathyroid hormone (PTH) could be of clinical utility for the disease's treatment and follow-up; however, PTH is not routinely determined due to limited available methods, often not fully validated in dogs. The aims of this study were the analytical validation of an immunoenzymatic method for the measurement of PTH in canine serum and the analysis of preliminary association of the obtained results with renal function. Twenty-six samples obtained from dogs healthy or affected with CKD were analysed. PTH was measured using a two-site immunoenzymometric human assay (ST AIA-PACK Intact PTH, Tosoh Bioscience). The analytical validation protocol evaluated the assay precision and accuracy. Also, the PTH's storage stability at 20 °C, 4 °C and -20 °C was assessed. Clinical validation was performed by comparing PTH values with creatinine, phosphorus and International Renal Interest Society (IRIS) stage. The method showed optimal precision and accuracy, whereas stability was adequate up to 4 h at 20 °C, 24 h at 4 °C and 6 months at -20 °C. PTH was positively associated with creatinine, phosphorus and IRIS stage. The investigated method was thus successfully validated in dogs, allowing its use for clinical purpose.
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http://dx.doi.org/10.3390/ani10122411DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7766972PMC
December 2020

Concordance between Histology, Immunohistochemistry, and RT-PCR in the Diagnosis of Feline Infectious Peritonitis.

Pathogens 2020 Oct 18;9(10). Epub 2020 Oct 18.

Department of Veterinary Medicine, University of Milan, 26900 Lodi, Italy.

Histology, immunohistochemistry (IHC), and reverse transcription polymerase chain reaction (RT-PCR) have been used to diagnose feline infectious peritonitis (FIP), but no information regarding the comparison of their diagnostic performances on the same organ is available. The aims of this study were to determine the concordance among these tests and to evaluate which combination of tests and organs can be used in vivo. Histology, IHC, and nested RT-PCR (RT-nPCR) for feline coronavirus (FCoV) were performed on spleen, liver, mesenteric lymph node, kidney, large and small intestine, and lung from 14 FIP and 12 non-FIP cats. Sensitivity, specificity, predictive values, likelihood ratios, and concordance were calculated. IHC and RT-nPCR had the highest concordance in lung and liver, histology and IHC in the other organs. The sensitivity of histology, IHC, and RT-nPCR on the different organs ranged from 41.7 to 76.9%, 46.2 to 76.9%, and 64.3 to 85.7%, respectively, and their specificity ranged from 83.3 to 100.0%, 100% and 83.3 to 100.0%. Therefore, IHC is recommended when histology is consistent with FIP. If RT-nPCR is performed as the first diagnostic approach, results should always be confirmed with IHC. Lung or liver provide accurate information regardless of the method, while IHC is preferred to RT-nPCR to confirm FIP in the kidney or intestine.
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http://dx.doi.org/10.3390/pathogens9100852DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7603141PMC
October 2020

Feline infectious peritonitis (FIP) and coronavirus disease 19 (COVID-19): Are they similar?

Transbound Emerg Dis 2021 Jul 20;68(4):1786-1799. Epub 2020 Oct 20.

Department of Veterinary Medicine, University of Milan, Lodi, Italy.

SARS-CoV-2 has radically changed our lives causing hundreds of thousands of victims worldwide and influencing our lifestyle and habits. Feline infectious peritonitis (FIP) is a disease of felids caused by the feline coronaviruses (FCoV). FIP has been considered irremediably deadly until the last few years. Being one of the numerous coronaviruses that are well known in veterinary medicine, information on FCoV could be of interest and might give suggestions on pathogenic aspects of SARS-CoV-2 that are still unclear. The authors of this paper describe the most important aspects of FIP and COVID-19 and the similarities and differences between these important diseases. SARS-CoV-2 and FCoV are taxonomically distant viruses, and recombination events with other coronaviruses have been reported for FCoV and have been suggested for SARS-CoV-2. SARS-CoV-2 and FCoV differ in terms of some pathogenic, clinical and pathological features. However, some of the pathogenic and immunopathogenic events that are well known in cats FIP seem to be present also in people with COVID-19. Moreover, preventive measures currently recommended to prevent SARS-CoV-2 spreading have been shown to allow eradication of FIP in feline households. Finally, one of the most promising therapeutic compounds against FIP, GS-441524, is the active form of Remdesivir, which is being used as one therapeutic option for COVID-19.
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http://dx.doi.org/10.1111/tbed.13856DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7537058PMC
July 2021

Haematological, serum biochemical and electrophoretic data on healthy captive Egyptian fruit bats ().

Lab Anim 2021 Apr 24;55(2):158-169. Epub 2020 Aug 24.

Department of Veterinary Medicine, University of Milan, Italy.

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 (). The aim of this study was to obtain haematological, biochemical and electrophoretic RIs from captive fruit bats. Blood was collected from 21 (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-, β-, β- 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 and, possibly, of other Chiroptera.
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http://dx.doi.org/10.1177/0023677220948542DOI Listing
April 2021

Serum paraoxonase 1 activity in cats: analytical validation, reference intervals, and correlation with serum amyloid A and alpha-1-acid glycoprotein.

J Vet Diagn Invest 2020 Nov 17;32(6):844-855. Epub 2020 Aug 17.

Department of Veterinary Medicine and Veterinary Teaching Hospital, University of Milan, Lodi, Italy.

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.
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http://dx.doi.org/10.1177/1040638720949638DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7649547PMC
November 2020

Flow cytometry expression pattern of CD44 and CD18 markers on feline leukocytes.

J Vet Diagn Invest 2020 Sep 28;32(5):706-709. Epub 2020 Jul 28.

Department of Veterinary Medicine (Martini, Bernardi, Giordano, Comazzi) and Veterinary Teaching Hospital (Martini, Bernardi, Giordano, Comazzi), University of Milan, Lodi, Italy.

The paucity of specific feline antibodies for flow cytometry (FC) is an ongoing challenge. Flow cytometrists must extrapolate information from relatively few markers. We evaluated the expression pattern of the panleukocyte markers CD18 and CD44 on leukocyte (white blood cell, WBC) subclasses in the peripheral blood (PB) of 14 healthy cats. The degree of expression of CD18 and CD44 was calculated as the ratio between the median fluorescence intensity (MFI) value of antibody-stained cells and autofluorescence. All samples were acquired with the same cytometer with constant photomultiplier setting and compensation matrices. Both molecules were expressed at higher levels on monocytes, intermediate levels on polymorphonuclear cells (PMNs), and lower levels on lymphocytes. CD18-MFI discriminated well among the 3 populations, whereas CD44-MFI mostly overlapped between monocytes and PMNs. However, CD44-MFI had a lower intra-population variability. Evaluation of CD18 and CD44, together with morphologic parameters, was useful for discriminating among WBC subclasses in healthy cats. This information may be helpful for future studies given that an increase in CD18-MFI may indicate reactive changes, whereas fluctuations in CD44-MFI may suggest neoplasia.
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http://dx.doi.org/10.1177/1040638720945670DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7488972PMC
September 2020

Comparison of simultaneous F-2-[18F] FDG PET/MR and PET/CT in the follow-up of patients with differentiated thyroid cancer.

Eur J Nucl Med Mol Imaging 2020 12 30;47(13):3066-3073. Epub 2020 Jun 30.

Department of Advanced Biomedical Sciences, University Federico II, Naples, Italy.

Aims: F-FDG PET/CT is the most accurate imaging modality in differentiated thyroid cancer (DTC) patients with either an aggressive histology, an absence of radioiodine uptake in neoplastic foci, or in the absence of imaging abnormalities in patients with an elevated serum thyroglobulin (Tg) level that progresses with time. We evaluated the diagnostic performance of FDG PET/MR in comparison with that of PET/CT.

Methods And Results: Following the injection of a single F-FDG activity, PET/MR and PET/CT were sequentially performed in 40 consecutive patients with DTC previously treated with total thyroidectomy and radioiodine ablation. All patients were then followed up for at least 6 months. PET/MR was positive in 11 patients and PET/CT in 10. PET/MR detected 33 tumor foci and PET/CT 30. During the follow-up of the 12 patients with negative initial PET studies and with a detectable serum Tg, only one patient had a neck recurrence and the administration of an empiric high activity of I in the other 11 patients did not reveal any tumor focus. In the 17 patients with an initial serum Tg level < 2 ng/mL, no recurrence occurred.

Conclusion: This study confirms the high diagnostic accuracy of FDG PET studies in DTC patients with elevated serum Tg levels and shows that PET/MR brings similar information as compared to PET/CT imaging.
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http://dx.doi.org/10.1007/s00259-020-04938-0DOI Listing
December 2020

A machine learning-based approach to directly compare the diagnostic accuracy of myocardial perfusion imaging by conventional and cadmium-zinc telluride SPECT.

J Nucl Cardiol 2020 May 18. Epub 2020 May 18.

Department of Advanced Biomedical Sciences, University Federico II, Via Pansini 5, 80131, Naples, Italy.

Background: We evaluated the performance of conventional (C) single-photon emission computed tomography (SPECT) and cadmium-zinc-telluride (CZT)-SPECT in a large cohort of patients with suspected or known coronary artery disease (CAD) and compared the diagnostic accuracy of the two systems using machine learning (ML) algorithms.

Methods And Results: A total of 517 consecutive patients underwent stress myocardial perfusion imaging (MPI) by both C-SPECT and CZT-SPECT. In the overall population, an excellent correlation between stress MPI data and left ventricular (LV) functional parameters measured by C-SPECT and by CZT-SPECT was observed (all P < .001). ML analysis performed through the implementation of random forest (RF) and k-nearest neighbors (NN) algorithms proved that CZT-SPECT has greater accuracy than C-SPECT in detecting CAD. For both algorithms, the sensitivity of CZT-SPECT (96% for RF and 60% for k-NN) was greater than that of C-SPECT (88% for RF and 53% for k-NN).

Conclusions: MPI data and LV functional parameters obtained by CZT-SPECT are highly reproducible and provide good correlation with those obtained by C-SPECT. ML approach showed that the accuracy and sensitivity of CZT-SPECT is greater than C-SPECT in detecting CAD.
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http://dx.doi.org/10.1007/s12350-020-02187-0DOI Listing
May 2020

Origin and transmission of Feline coronavirus type I in domestic cats from Northern Italy: a phylogeographic approach.

Vet Microbiol 2020 May 12;244:108667. Epub 2020 Apr 12.

L. Sacco Department of Biomedical and Clinical Sciences, University of Milan, Via G. B. Grassi 74, 20157 Milan, Italy.

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 subs/site/year (95% HPD: 1.3-3.7 × 10), 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.
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http://dx.doi.org/10.1016/j.vetmic.2020.108667DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7152909PMC
May 2020

Relationship between heart rate response and cardiac innervation in patients with suspected or known coronary artery disease.

J Nucl Cardiol 2020 Mar 12. Epub 2020 Mar 12.

Department of Advanced Biomedical Sciences, University Federico II, Via Pansini 5, 80131, Naples, Italy.

Background: Chronotropic response to pharmacological stress test is blunted in patients with autonomic neuropathy. The relationship between heart rate (HR) changes during pharmacological stress test and cardiac autonomic dysfunction has not been fully investigated. We assessed the potential interplay between HR response (HRR) and myocardial innervation in patients with suspected or known coronary artery disease (CAD).

Methods And Results: We studied 71 patients with suspected or known CAD referred to pharmacological stress myocardial perfusion imaging and I metaiodobenzylguanidine (I-MIBG) cardiac scintigraphy. HRR was calculated as the maximum percent change from baseline according to the formula: (peak HR - rest HR)/rest HR × 100. I-MIBG heart-to-mediastinum (H/M) ratio was calculated and a late H/M ratio < 1.6 was considered abnormal. HRR progressively decreased with decreasing late H/M ratio (P for trend = 0.02) and a significant correlation between HRR and late H/M ratio (P = 0.03) was observed. The addition of HRR to a model including age, diabetes, known CAD, left ventricular ejection fraction, and stress-induced ischemia added incremental value in predicting an abnormal late H/M ratio, increasing the global chi-square from 8.09 to 13.8 (P = 0.02).

Conclusions: The relationship between HRR and cardiac sympathetic innervation in patients with suspected or known CAD confirms a strong interplay between cardiac response to stress tests and cardiac autonomic activation. This finding suggests that HRR may be used as a surrogate for assessing cardiac sympathetic function.
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http://dx.doi.org/10.1007/s12350-020-02091-7DOI Listing
March 2020

Paraoxonase-1 activity evaluation as a diagnostic and prognostic marker in horses and foals.

J Vet Intern Med 2020 Mar 10;34(2):949-954. Epub 2020 Mar 10.

Department of Veterinary Sciences, University of Pisa, Pisa, Italy.

Background: In several species, paraoxonase-1 (PON-1) decreases during inflammation, because of the presence of oxidative stress; its measurement recently has been validated in horses, but its role as a clinical biomarker is unknown.

Objectives: To evaluate sensitivity, specificity and likelihood ratio of PON-1 activity to identify systemic inflammatory response syndrome (SIRS)-positive horses or horses with a poor prognosis.

Animals: One hundred seventy-two blood samples from 58 sick horses from 3 different veterinary hospitals.

Methods: In a cross-sectional study, PON-1 activity was measured upon admission and at 24-hour intervals until discharge or death, and results were analyzed based on SIRS status and outcome.

Results: No statistically significant difference was found in median PON-1 activity between SIRS and non-SIRS cases or between survivors and non-survivors except for mares, in which PON-1 activity was significantly lower in SIRS-positive horses (P = .05). The sensitivity of PON-1 activity in identifying horses with SIRS or negative outcome was low (0.0%-46.2% depending on the examined group) but its specificity was high (87.0%-100.0%). However, when PON-1 is low, the likelihood of death is 2.40-3.89 times higher than the likelihood of survival. Repeated measurement of PON-1 after treatment does not predict outcome.

Conclusions And Clinical Importance: Evaluation of PON-1 activity in horses with inflammation might be advisable in the future, but only low activity at admission may be relevant in predicting SIRS or negative outcome.
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http://dx.doi.org/10.1111/jvim.15722DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7096640PMC
March 2020

Low-dose dynamic myocardial perfusion imaging by CZT-SPECT in the identification of obstructive coronary artery disease.

Eur J Nucl Med Mol Imaging 2020 07 17;47(7):1705-1712. Epub 2019 Dec 17.

Department of Advanced Biomedical Sciences, University Federico II, Naples, Italy.

Background: We measured myocardial blood flow (MBF) and myocardial perfusion reserve (MPR) by a dynamic low-dose CZT-SPECT protocol in patients with suspected or known coronary artery disease (CAD) and investigated the capability of dynamic data in predicting obstructive CAD. A total of 173 patients with suspected or known CAD underwent dynamic CZT-SPECT after the injection of 155 MBq and 370 MBq of Tc-sestamibi for rest and stress imaging, respectively. Standard rest and stress imaging were performed at the end of each dynamic scan. A total perfusion defect (TPD) < 5% were considered normal. Obstructive CAD was defined as ≥ 70% stenosis at coronary angiography.

Results: Global MPR was lower (p < 0.05) in patients with abnormal compared with those with normal MPI (2.40 ± 0.7 vs. 2.70 ± 0.8). A weak, albeit significant correlation between TPD and MPR (r = - 0.179, p < 0.05) was found. In 91 patients with available angiographic data, hyperemic MBF (2.59 ± 1.2 vs. 3.24 ± 1.1 ml/min/g) and MPR (1.96 ± 0.7 vs. 2.74 ± 0.9) were lower (both p < 0.05) in patients with obstructive CAD (n = 21) compared with those without (n = 70). At univariable analysis, TPD, hyperemic MBF, and MPR were significant predictors of obstructive CAD, whereas only MPR was independent predictor at multivariable analysis (p < 0.05). At per vessels analysis, regional hyperemic MBF (2.59 ± 1.2 vs. 3.24 ± 1.1 ml/min/g) and regional MPR (1.96 ± 0.7 vs. 2.74 ± 0.9) were lower in the 31 vessels with obstructive CAD compared with 242 vessels without (both p < 0.05).

Conclusions: In patients with suspected or known CAD, MPR assessed by low-dose dynamic CZT-SPECT showed a good correlation with myocardial perfusion imaging findings and it could be useful to predict obstructive CAD.
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http://dx.doi.org/10.1007/s00259-019-04644-6DOI Listing
July 2020

Feline gut microbiota composition in association with feline coronavirus infection: A pilot study.

Res Vet Sci 2019 Aug 9;125:272-278. Epub 2019 Jul 9.

Department of Veterinary Medicine, University of Milan, Via Celoria 10, 20122 Milano, MI, Italy; Veterinary Teaching Hospital, University of Milan, via dell'Università, 6, 26900 Lodi, LO, Italy.

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.
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http://dx.doi.org/10.1016/j.rvsc.2019.07.003DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7111766PMC
August 2019

Validation of a paraoxon-based method for measurement of paraoxonase (PON-1) activity and establishment of RIs in horses.

Vet Clin Pathol 2018 Mar;47(1):69-77

Department of Veterinary Sciences, University of Pisa, San Piero a Grado, Italy.

Background: Paraoxonase-1 (PON-1) is an antioxidant compound that is considered a negative acute phase protein. No information on the analytic performance of the paraoxon method for measuring PON-1 in horse serum is available.

Objectives: The aim of this study was to validate a paraoxon-based method to measure PON-1 in horses and to establish RIs in healthy horses and foals.

Methods: Horses and foals classified as healthy after physical examination and routine biochemistry were used in the study. Serum PON-1 activity was measured with an automated spectrophotometer and an enzymatic method validated in other species. After the analytic validation (precision, accuracy, interference studies), RIs were determined using the Reference Value Advisor software. The possible sex-, age-, and breed-related differences were statistically investigated.

Results: The healthy study population included 120 horses and 55 foals. The paraoxon-based method was precise (CVs < 4.0%) and accurate (P < .001 in linearity under dilution and spike-recovery testing) but was affected by interference from mild bilirubinemia, severe lipemia, and hemoglobinemia. The RIs recorded in the whole population were 38.1-80.8 U/mL. According to the Harris and Boyd test, it would be advisable to use separate RIs only for adult females and for Warmblood and Trotter adults.

Conclusions: This study demonstrated that the analytic performances of the paraoxon-based method for measurement of PON-1 in horses are acceptable. The PON-1 activity is lower in horses than in other domestic species. These results may provide a basis for further studies designed to establish whether healthy and sick horses can be correctly classified by using the PON-1 assay.
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http://dx.doi.org/10.1111/vcp.12562DOI Listing
March 2018

Comparison of the performance of laboratory tests in the diagnosis of feline infectious peritonitis.

J Vet Diagn Invest 2018 May 11;30(3):459-463. Epub 2018 Feb 11.

Department of Veterinary Medicine, University of Milan, Milan, Italy; Central Laboratory, Veterinary Teaching Hospital, University of Milan, Lodi, Italy.

We compared the performance of clinicopathologic and molecular tests used in the antemortem diagnosis of feline infectious peritonitis (FIP). From 16 FIP and 14 non-FIP cats, we evaluated retrospectively the sensitivity, specificity, and likelihood ratios (LRs) of serum protein electrophoresis, α-acid glycoprotein (AGP) on peripheral blood, screening reverse-transcription nested PCR (RT-nPCR) on the 3'-untranslated region (3'-UTR), and spike (S) gene sequencing on peripheral blood, body cavity effusions, and tissue, as well as body cavity cytology and delta total nucleated cell count (ΔTNC). Any of these tests on blood, and especially the molecular tests, may support or confirm a clinical diagnosis of FIP. A negative result does not exclude the disease except for AGP. Cytology, 3'-UTR PCR, and ΔTNC may confirm a clinical diagnosis on effusions; cytology or 3'-UTR PCR may exclude FIP. Conversely, S gene sequencing is not recommended based on the LRs. On tissues, S gene sequencing is preferable when histology is highly consistent with FIP, and 3'-UTR PCR when FIP is unlikely. Combining one test with high LR+ with one with low LR- (e.g., molecular tests and AGP on blood, ΔTNC and cytology in effusions) may improve the diagnostic power of the most used laboratory tests.
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http://dx.doi.org/10.1177/1040638718756460DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6505812PMC
May 2018

Reference intervals for hematological and biochemical parameters, acute phase proteins and markers of oxidation in Holstein dairy cows around 3 and 30days after calving.

Res Vet Sci 2017 Oct 20;114:322-331. Epub 2017 Jun 20.

Department of Veterinary Medicine, University of Milan, via Celoria 10, 20133 Milan, Italy; Central Laboratory, Veterinary Teaching Hospital, University of Milan, via dell'Università 6, 26900 Lodi, Italy.

The establishment of specific reference intervals (RIs) is advisable when different metabolic conditions in the patient population are present or when existing RIs are not suitable. Holstein dairy cows, at the start of lactation, experience a negative energy balance and an altered immune function that can lead to different levels of blood parameters compared with cows at peak lactation. The aim of this study was the determination of RIs for Holstein cows at 3±1 and 30±3days in milk. To this aim, 145 cows, from 4 herds, were sampled. A wide panel of hematological and biochemical analytes was determined, as well as the measurement of markers of inflammation/oxidation. RIs were generated following the American Society of Veterinary Clinical Pathology (ASVCP) guidelines and the effects of lactation period, parity, herd and day of sampling were determined. Data from 39 out of 52 analytes were significantly different according to lactation period. Data of 13 out of 52 analytes were not significantly different between the two groups. Some differences were also recorded between herds and parity groups but the adoption of specific RIs for these subgroups, however, has practical limitations. In conclusion the use of RIs specific for the lactation period is highly justified from both a statistical and a biological point of view. Pre-analytical factors associated with parity or management need to be considered in the evaluation of results of some analytes.
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http://dx.doi.org/10.1016/j.rvsc.2017.06.012DOI Listing
October 2017

Colorimetric and electrophoretic evaluation of lipoprotein fractions in healthy neonatal calves: Comparison with results from adult cows and from calves with inflammatory conditions.

Res Vet Sci 2017 Apr 13;111:108-112. Epub 2017 Feb 13.

Department of Veterinary Medicine, University of Milan, Via Celoria, 10-20133 Milan, Italy; Central Laboratory, Veterinary Teaching Hospital - University of Milan, Via dell'Università, 6, 26900 Lodi, Italy.

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±11.6mg/dL) and an increase of VLDL% (12.1% ±13.1%) compared with controls (77.6%±8.6%; 41.5±11.2mg/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.
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http://dx.doi.org/10.1016/j.rvsc.2017.02.007DOI Listing
April 2017

Diagnosing feline infectious peritonitis using the Sysmex XT-2000iV based on frozen supernatants from cavitary effusions.

J Vet Diagn Invest 2017 May 16;29(3):321-324. Epub 2017 Feb 16.

Department or Veterinary Medicine, University of Milan, Milan, Italy.

The delta total nucleated cells (ΔTNC) measurement with the Sysmex XT-2000iV (Sysmex Europe, Norderstedt, Germany) has high diagnostic accuracy on effusions in feline infectious peritonitis (FIP) cases, but the test can be performed only on fresh samples. We evaluated whether supernatants from effusions retain the ability to induce cell clumping and assessed the diagnostic accuracy of this modified ΔTNC method. Effusions were collected from FIP cats ( n = 19) and from cats with other diseases ( n = 15). ΔTNC was measured on fresh samples and on frozen-thawed supernatants after the addition of feline blood at 1:10 dilution. Diagnostic accuracy was assessed at the cutoffs of suggestive of FIP (ΔTNC = 1.7) and consistent with FIP (ΔTNC = 3.4). The influence of the protein content, number of added cells, and magnitude of dilution were also investigated. Specificity and positive predictive value were 100% for both the methods. Sensitivity and negative predictive value were higher for the modified ΔTNC (84.2% and 83.3%, respectively, at the cutoff of 1.7; 78.9% and 78.9%, respectively, at the cutoff of 3.4) than for the ΔTNC on fresh samples (78.6% and 81.3%, respectively, at the cutoff of 1.7; 57.1% and 68.4%, respectively, at the cutoff of 3.4). Protein content, total cell count of the added blood, and magnitude of dilutions did not influence the results. Supernatants of frozen effusions from FIP cats retain the ability to induce cell clumping, thus the modified ΔTNC measurement is a reliable tool to diagnose FIP on samples that cannot be analyzed immediately.
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http://dx.doi.org/10.1177/1040638716689115DOI Listing
May 2017

Reverse transcriptase loop-mediated isothermal amplification for the detection of feline coronavirus.

J Virol Methods 2017 05 18;243:105-108. Epub 2017 Jan 18.

Department of Veterinary Medicine, University of Milan, Milan, Italy; Veterinary Teaching Hospital, University of Milan, Milan, Italy.

The Feline coronavirus (FCoV) is the etiological agent of feline infectious peritonitis (FIP), a lethal disease of felids. The role of molecular methods is controversial for the diagnosis of FIP, while essential for the identification of the shedders. Thus, a fast and inexpensive method for the detection of FCoV could be beneficial, especially in multicat environments. A reverse transcription loop mediated isothermal amplification (RT-LAMP) assay was developed. RNA extraction and RT-nPCR for FCoV were performed on thirty-two samples (11 faeces, 9 blood, 8 effusions, and 4 lymph nodes) collected from 27 cats. Six RT-LAMP primers were designed from the same conserved region of RT-nPCR, and the assay was run at 63°C for one hour. Results were evaluated through both agarose gel run and hydroxynapthol blue (HNB) dye and then compared with RT-nPCR results for the assessment of sensitivity and specificity. The overall specificity was 100%, but the sensitivity was 50% and 54.5% for agarose gel and HNB respectively. Therefore, RT-LAMP seems optimal to confirm the presence of the virus, but not applicable to exclude it.
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http://dx.doi.org/10.1016/j.jviromet.2017.01.009DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7113849PMC
May 2017

Serum creatine kinase isoenzymes and macroenzymes in dogs with different neurologic diseases.

Vet Clin Pathol 2017 Mar 13;46(1):91-99. Epub 2017 Jan 13.

Portoni Rossi Veterinary Hospital, Zola Predosa, Italy.

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.
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http://dx.doi.org/10.1111/vcp.12443DOI Listing
March 2017

Sensitivity and specificity of manual and automated measurements of reticulocyte parameters for classification of anemia in dogs: 174 cases (1993-2013).

J Am Vet Med Assoc 2016 Oct;249(7):776-86

OBJECTIVE To assess sensitivity and specificity of manual and automated measurements of reticulocyte percentage, number, and production index for classification of anemia in dogs. DESIGN Retrospective case series SAMPLE 174 blood smears from client-owned dogs with anemia collected between 1993 and 2013 for which reticulocyte parameters were determined manually (nonregenerative anemia, 22; preregenerative anemia, 23; regenerative anemia, 28) or with an automated laser-based counter (nonregenerative anemia, 66; preregenerative anemia, 17; regenerative anemia, 18). PROCEDURES Diagnostic performance was evaluated with receiver operating characteristic (ROC) curves by considering preregenerative anemia as nonregenerative or regenerative. Sensitivity, specificity, and positive likelihood ratio were calculated by use of cutoffs determined from ROC curves or published reference limits. RESULTS Considering preregenerative anemia as non regenerative, areas under the curve (AUCs) for reticulocyte percentage, number, and production index were 97%, 93%, and 91% for manual counting and 93%, 90%, and 93% for automated counting. Sensitivity, specificity, and positive likelihood ratio were 82% to 86%, 82% to 87%, and 4.6 to 6.4, respectively. Considering preregenerative anemia as regenerative, AUCs were 77%, 82%, and 80% for manual counting and 81%, 82%, and 92% for automated counting. Sensitivity, specificity, and positive likelihood ratio were 72% to 74%, 76 to 87%, and 2.7 to 6.2, respectively. CONCLUSIONS AND CLINICAL RELEVANCE Whereas all reticulocyte parameters identified regeneration in anemic dogs, the performance of specific parameters was dependent on the method used. Findings suggested that lower cutoffs than published reference limits are preferred for reticulocyte number and production index and higher cutoffs are preferred for reticulocyte percentage. Reticulocyte production index may be useful when the pretest probability of regeneration is moderate.
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http://dx.doi.org/10.2460/javma.249.7.776DOI Listing
October 2016
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