Canine Parvoviruses. Rapid Diagnostic by Transmission Electron Microscopy and Histopathology Techniques

Authors: Catroxo, M.H.B.; Del Fava, C.; Martins, A.M.C.R.P.F.
DOI
10.25125/ijoear-mar-2026-17
DIN
IJOEAR-MAR-2026-17
Abstract

Canine parvovirus infection is a highly contagious disease of significant clinical and epidemiological relevance in veterinary medicine. It carries a high risk of severity and lethality, posing a particular threat to young, unvaccinated, or immunocompromised animals, and is one of the leading causes of severe gastroenteritis and mortality in dogs. Canine parvovirus (CPV-2) belongs to the family Parvoviridae and the genus Protoparvovirus. Two distinct parvoviruses are known to infect dogs: CPV-1, also called the minute virus of canines (MCV), and the pathogenic CPV-2. MCV may cause pneumonia, myocarditis, and enteritis in young pups, or transplacental infections in pregnant dams, leading to embryo resorption and fetal death. CPV-2, the causative agent of acute hemorrhagic enteritis and myocarditis in dogs, is one of the most important pathogenic viruses, with high morbidity (100%) and frequent mortality—up to 10% in adult dogs and 91% in puppies. This study aimed to diagnose canine parvovirus in fecal samples, rectal swabs, and organ fragments from dogs using transmission electron microscopy and histopathological techniques. Between 1995 and 2016, approximately 665 fecal specimens or small intestine fragments from dogs with diarrhea were sent to the Electron Microscopy Laboratory of the Biological Institute of São Paulo, SP, Brazil, for viral diagnosis. The samples were processed using transmission electron microscopy (negative staining, immunoelectron microscopy, immunocytochemistry with colloidal gold labeling, and resin embedding) and routine histopathological techniques. Using a Philips EM 208 transmission electron microscope, all samples were analyzed by the negative staining technique. In 62 samples (9.32%), a large number of parvovirus particles were observed—non-enveloped, isometric, and characterized as "complete" and "empty," measuring approximately 20 nm in diameter. Positive results in immunoelectron microscopy were confirmed by the presence of aggregates formed through antigen-antibody interactions. In immunocytochemistry, the antigen-antibody reaction was strongly enhanced by dense colloidal gold particles in all 62 positive samples. Histopathological analysis revealed hemorrhagic small intestine with villous necrosis, multiple hepatic lobules with moderate vacuolar degeneration of hepatocytes, kidneys with extensive areas of cortical coagulative necrosis, severe pulmonary edema, and moderate splenic white pulp reaction.

Keywords
Clinical cases Anatomopathology Dog disease
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Introduction

Canine parvovirus infection is a highly contagious disease with significant severity and lethality, posing a major threat particularly to young animals. It is one of the leading causes of severe gastroenteritis and mortality in unvaccinated and immunocompromised dogs. The disease occurs worldwide in domestic dogs and other members of the Canidae family, and outbreaks have been reported in several countries, including Italy (Mira et al., 2024), Thailand (Charoenkul et al., 2024), Turkey (Ulas et al., 2024), Egypt (Magouz et al., 2023), China (Fu et al., 2022), the United States (Hong et al., 2007), Chile (Castillo et al., 2020), and India (Abhiram et al., 2023).

In Brazil, thousands of animals are infected every year, with numerous cases reported in different states, such as Rio de Janeiro (Castro et al., 2007), Rio Grande do Sul (Oliveira et al., 2018), Paraíba (Souto et al., 2018), Minas Gerais (Silva et al., 2022), and São Paulo (Cappellaro et al., 1995; Catroxo et al., 2015).

Canine parvovirus (CPV-2) belongs to the family Parvoviridae and the genus Protoparvovirus (Khatri et al., 2017). These viruses are isometric, non-enveloped, measuring 18–26 nm in diameter, with icosahedral symmetry and 32 capsomers surrounding a core containing single-stranded DNA approximately 5.2 kb in length (Bennett et al., 2008). Parvoviruses encode two nonstructural proteins, NS1 and NS2. NS1 is associated with nuclear processes required for viral replication (Moon et al., 2013). A recent study demonstrated that NS2 interacts with chromatin, regulating cellular proteins (Mattola et al., 2022).

Two distinct parvoviruses are known to infect dogs: CPV-1, also known as minute virus of canine (MCV), and the pathogenic CPV-2 (Nandy & Kumar, 2010). MCV may cause pneumonia, myocarditis, and enteritis in young puppies, as well as transplacental infections in pregnant dams, leading to embryonic resorption and fetal death (Carmichael et al., 1994). CPV-2, the causative agent of acute hemorrhagic enteritis and myocarditis in dogs, is one of the most important pathogenic viruses, with high morbidity (up to 100%) and frequent mortality rates reaching up to 10% in adult dogs and 91% in puppies (Nandy & Kumar, 2010).

Canine parvovirus is transmitted through oral contact with infected feces or contaminated surfaces, such as soil, shoes, dog toys, and other fomites (Decaro & Buonavoglia, 2012). Following an incubation period averaging 4 to 14 days, the virus replicates in the crypts of the small intestine, causing destruction of enterocytes, rupture of the mucosal barrier, and atrophy of intestinal villi. Affected animals may shed the virus in feces for more than 10 days.

The acute disease begins with depression, anorexia, high fever, profuse vomiting, and severe diarrhea. The diarrhea is abundant, often containing mucus and/or blood, and dehydration develops rapidly. The clinical scenario has become more complex due to the emergence of several variants over the years, including CPV-2a, CPV-2b, new CPV-2a, new CPV-2b, and CPV-2c, as well as the involvement of domestic and wild canids, causing serious damage to kennels (Nandy & Kumar, 2010).

The main histopathological alterations include atrophy of intestinal villi and dilation of crypts due to selective viral replication in enterocytes, bronchiolar epithelial necrosis in the lungs, vacuolar degeneration of hepatocytes, kidneys with areas of necrosis, and lymphoplasmacytic myocarditis in the heart (Kumari et al., 2020; Al-Bayati et al., 2016; Decaro & Buonavoglia, 2012; Fagbohun et al., 2020).

In breeding kennels and animal shelters, parvovirosis represents a frequent veterinary concern due to the rapid spread of the virus, resulting in significant animal losses and economic damage. Rapid diagnosis is essential, as the disease favors the development of secondary infections, accelerating clinical progression. Death in unvaccinated animals or those with ineffective vaccination protocols may occur within 2 to 3 days after the onset of clinical signs (Santana et al., 2019).

The objective of this study was to diagnose canine parvovirus in fecal samples, rectal swabs, and organ fragments from dogs, using transmission electron microscopy and histopathological techniques.

Conclusion

This study successfully demonstrated the utility of transmission electron microscopy techniques—including negative staining, immunoelectron microscopy, immunocytochemistry with colloidal gold labeling, and resin embedding—combined with histopathological analysis for the rapid and accurate diagnosis of canine parvovirus infection. Of the 665 samples analyzed, 62 (9.32%) were positive for CPV-2, with the highest prevalence observed in animals up to 11 months of age. Co-infections with coronavirus, paramyxovirus, and Mycoplasma spp. were identified in a significant proportion of cases. The ultrastructural and histopathological findings were consistent with the characteristic tropism of parvovirus for rapidly dividing cells, particularly in the intestinal crypts, and revealed systemic involvement including hepatic, renal, pulmonary, and splenic alterations. The combination of these diagnostic techniques provides reliable support for clinical diagnosis and contributes to the understanding of disease pathogenesis. Rapid diagnosis remains essential for effective clinical management and implementation of control measures, while preventive vaccination continues to be the primary strategy for reducing the impact of this significant veterinary disease.

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