README file
Data Set Title: “Dataset of animal and environment sampling tested for Canine parvovirus and Feline panleukopenia virus in a Veterinary Teaching Hospital”
Dataset authors: Veronica Facile (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna); 
Laura Gallina (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna);
Ilara Magagnoli (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna);
Massimo Giunti (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna);
Mara Battilani (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna);
Andrea Balboni (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna);

Data Set Contact Persons: Andrea Balboni (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna), email: a.balboni@unibo.it; Mara Battilani (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna), email: mara.battilani@unibo.it; Laura Gallina (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna), email: laura.gallina@unibo.it; Veronica Facile (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna), email: veronica.facile2@unibo.it 

Data Set License: this data set is distributed under a Creative Commons Attribution 4.0 International (CC BY) license, https://creativecommons.org/licenses/by/4.0/  

Year of completion: 2022
Year of publication: 2025

Data set Contents
The data set consists of:
•	1 tabular qualitative file in .xlsx format
DatasetViralSurveillance_2026.xlsx
•	1 readme file in .rtf format
DatasetViralSurveillance_2026_README.rtf

Data set Documentation
Abstract
The dataset contains in sheet labeled as “Animal” signalment and diagnostic data of a population of 5 dogs and 2 cats and in sheet labeled as “Environment” 21 environmental sampling points for each animal include, tested for the presence of feline panleukopenia virus and canine parvovirus DNA at the Veterinary Teaching Hospital of the Department of Veterinary Medical Science of the University of Bologna.

Content of the files 
file DatasetViralSurveillance_2026.xlsx contains in sheet “Animal” signalment and diagnostic data on 5 dogs and 2 cats tested for the presence of canine parvovirus and feline panleukopenia virus DNA at the Veterinary Teaching Hospital of the Department of Veterinary Medical Science of the University of Bologna. Specifically, information about the breed, the origin, the age, the sex, the vaccination status, the outcome, the number of day of hospitalization, the dates of sampling and sampling sites and the qPCR, genotyping and viral isolation results are indicated for each tested animal. In sheet “Environment” 21 environmental sampling points for each animal included, tested for the presence of canine parvovirus and feline panleukopenia virus DNA at the Veterinary Teaching Hospital of the Department of Veterinary Medical Science of the University of Bologna, are reported. Specifically, qPCR results for each sampling point before and after disinfection procedures are reported for each animal hospitalization.

Methodologies 
The sampling was performed using swabs: dry for rectal sampling and pre-soaked in phosphate saline buffer for haircoat and environmental sampling. The qPCR assay used in this study was previously described by Balboni and colleagues (2018)1. Reactions were performed using the PowerUp SYBR Green Master Mix (Life Technologies, Thermo Fisher Scientific, Waltham, MA, USA) and using the Real Time StepOnePlus™ system (Applied Biosystem, Carlsbad, CA, USA) with a total reaction volume of 20 μl per tube. Each tube contained 10 μl Master Mix reagents, 1.5 μl of forward and reverse primers with a concentration of 10µM each, 2 μl of DNA template, and 5.6 μl of water for molecular biology. The thermal cycling protocol consisted of an initial denaturation at 95°C for 5 minutes, followed by 45 cycles of denaturation at 95°C for 15 seconds and annealing/extension at 60°C for 1 minute. Melting experiment for the evaluation of the specificity of the reaction was performed after the last extension step by a continuous increment from 55 ◦C to 95 °C, and specific melting temperatures (Tm) were about 76.5 °C for CPV-2 and 77.5 °C for FPV. Viruses identified in each animal (RS) were genetically characterized by analysis of the VP2 gene using a previously described end-point PCR (Balboni et al., 2021)2 that was carried out using the Phusion Hot Start II High-Fidelity DNA Polymerase Kit (Thermo Fisher Scientific, Life Technologies, Waltham, MA, USA) according to the manufacturer’s instructions. Amplicons of the expected size obtained with the PCR assay carried out in this study were sequenced by Sanger method using both forward and reverse primers.
Viral isolation on suitable cellular substrate (Canine fibrosarcoma cell line-A72 for CPV and Crandell-Rees Feline Kidney cell line for FPV), was performed on haircoat samples. The inoculum was prepared by eluting the haircoat swab in a tube containing 2 mL of Dulbecco’s Modified Eagle Medium (DMEM, Gibco, Thermo Fisher Scientific, Waltham, MA, USA)  supplemented with 10% antibiotic-antimycotic (100X, Gibco, Thermo Fisher Scientific, Waltham, MA, USA), vortexed for 30 s, inverted and centrifugated at 1000 ×g for 15 min. The supernatant was collected, filtered (0.22 µm syringe filters – VWR®, Avantor, Radnor, PA, USA) and dispended in sterile tubes.
Following inoculum preparation, three serial passages in cell culture were performed prior to assessing viral growth using qPCR for quantification of parvoviral DNA.
The infection was performed on six-well plates, each well was inoculated with 1.5 mL of inoculum prepared for each sample and the plate was incubated at 37°C in a humidified 5% CO₂ incubator for 72 hours. The plates were frozen and thawed three times and the supernatants collected to perform three consecutive passages in cell culture for each sample and, after DNA extraction, molecular testing was performed.

 Balboni, A., Bassi, F., De Arcangeli, S., Zobba, R., Dedola, C., Alberti, A., Battilani, M., 2018. Molecular analysis of carnivore Protoparvovirus detected in white blood cells of naturally infected cats. BMC Vet. Res. 4, 41. https://doi.org/10.1186/s12917-018-1356-9. 

2 Balboni A, Niculae M, Di Vito S, Urbani L, Terrusi A, Muresan C, Battilani M. The detection of canine parvovirus type 2c of Asian origin in dogs in Romania evidenced its progressive worldwide diffusion. BMC Vet Res. 2021 Jun 5;17(1):206. doi: 10.1186/s12917-021-02918-6.