key: cord-0010552-bqemkljl
authors: Klingeborn, B.; Moreno‐López, J.
title: Diagnostic Experience from an Epidemic of Canine Parvoviral Enteritis
date: 2010-05-13
journal: J Vet Med B Infect Dis Vet Public Health
DOI: 10.1111/j.1439-0450.1980.tb01794.x
sha: a6185cbe5dbf3f94f697b4d21643d69fd34eb209
doc_id: 10552
cord_uid: bqemkljl

SUMMARY: Hemagglutination (HA) test and electron microscopy (EM) were used to diagnose canine parvoviral enteritis on fecal samples from 58 hospitalized dogs of a huge epidemic. By HA‐tests the presence of the canine parvovirus (CPV) involved was shown in samples from 15 dogs (∼ 26%). A reference antiserum was used to identify the virus of each sample by hemagglutination inhibition (HI). By EM parvovirus‐like particles were seen in samples from 28 dogs (∼ 48%). Particles in six samples were identified as CPV by immunoaggregation. Sera from 17 out of 19 dogs examined showed specific HI titers during the acute stage of illness. A high incidence of concomitant infection with Campylobacter spp was found by the bacteriologists. ZUSAMMENFASSUNG: Diagnostische Erfahrungen während einer Epidemie von Parvovirus‐Enteritis der Hunde Während einer grossen Epidemie von Parvovirus‐Enteritis der Hunde (in Schweden) wurde der Nachweis von Virus in Kotproben mit Hilfe der Hëmagglutination (HA) und Elektronenmikroskopie (EM) versucht; die Kotproben stammten von 58 hospitalisierten Hunden. Mit dem HA‐Test ließ sich das ursächliche Parvovirus in den Proben von 15 Hunden nachweisen (= ∼ 26%). In jeder von diesen Proben wurde das Virus durch Hämagglutination‐Hemmung (HAH) mit einem Referenzserum identifiziert. Mit der EM waren Parvovirus‐ähnliche Partikel in den Proben von 28 Hunden zu finden (= ∼ 48%). In 6 von diesen Proben wurden die Partikel mittels der Immunaggregation als das ursächlichen Parvovirus identifiziert. In 17 Serumproben von 19 untersuchten Hunden wurden spezifische HAH‐Titer während der akuten Krankheitsphase festgestellt. Die Bakteriologen wiesen ein gehäuftes Vorkommen von gleichzeitiger Infektion mit Campylobacter spp nach. RÉSUMÉ: Expériences de diagnostic durant une épidémie d'entérite à Parvovirus chez des chiens On a recherché la mise en évidence du virus dans des matières fécales à l'aide de l'hémagglutination (HA) et de la microscopie électronique (EM) durant une forte épidémie d'entérite à Parvovirus chez des chiens en Suède. Les échantillons d'excréments provenaient de 58 chiens hospitalisés. Le Parvovirus a été mis en évidence par test HA dans les échantillons de 15 chiens (∼ 26%). Le virus a été identifié dans chacun de ces échantillons au moyen de l'inhibition de l'hémagglutination (HAH) avec un sérum de référence. On a trouvé des particules identiques au microscope électronique dans les échantillons de 28 chiens (∼ 48%). Les particules dans 6 de ces prélèvements furent identifiées comme Parvovirus au moyen de l'immunoaggrégation. Un titre HAH spécifique a été établi durant la phase aiguë de la maladie dans 17 échantillons sérologiques sur 19 chiens examinés. La bactériologie a montré la présence fréquente d'une infection simultanée à Campylobacter spp. RESUMEN: Experiencias en el diagnóstico de una epidemia de parvovirus‐enteritis en caninos Mediante pruebas de hemaglutinación y microscopía electrónica se diagnosticó parvovirus‐enteritis en caninos en muestras fecales de 58 caninos hospitalizados a raiz de una extensa epidemia. Con la prueba de hemaglutinación se detectó la presencia de virus en muestras fecales de 15 caninos (∼ 26%). Un antisuero de referencia se utilizó para la identificación del virus (prueba de inhibición de la hemaglutinación). Con microscopía electrónica se identificó parvovirus en muestras fecales de 28 caninos (∼ 48%). El virus fué identificado en seis muestras fecales utilizando la prueba de inmunoagregación. Sueros de 17 de 19 caninos examinados demostraron títulos específicos de inhibición de la hemaglutinación durante la etapa aguda de la enfermedad. Se demostró también una alta incidencia de infección concomitante con Campylobacter spp.

In 1978 a new disease of dogscanine parvoviral enteritisemerged in some countries with first exhaustive descriptions from the United States (1, 4, 7) . Even if, at the beginning, a coronavirus has been found in feces ( l ) , the predominant virus was a parvovirus, serologically different from the "minute virus of canines (MCV)" ( 6 ) , also a parvovirus known since 1970 (3) .

The new disease of dogs has similarities to feline panleukopenia (1, 7) , and the canine parvovirus (CPV) involved was found to be serologically closely related to feline panleukopenia and mink enteritis virus (2, 4). It is anticipated that the CPV is a mutant of feline panleukopenia virus (6) , but a comparative analysis of these viruses has yet to be made.

In the winter months of 1979/80 a huge epidemic of canine enteritis occurred in Sweden in connection with a dog exhibition, attadring mainly young dogs. Thousands of dogs were taken ill. The epidemic was suspected to be canine parvoviral enteritis and therefore feces and serum samples were submitted to virological examination. The rapid diagnosis was favored following CARMICHAEL'S procedure of detecting the CPV hemagglutinin in feces and the antihemagglutinin in serum (6) . To this we added electron microscopy and immune electron microscopy of particles in fecal samples, immunodiffusion and counterimmunoelectrophoresis. The present report is a summary of experience using these procedures. 

All the dogs examined were hospitalized in two animal clinics. Most of these dogs showed signs of severe gastroenteritis, i. e., vomiting, anorexia, diarrhea and rapid dehydration. On the basis of these signs and lymphocytopenia the disease was suspected to be canine parvoviral enteritis.

The feces and/or serum samples were collected during the acute stage of illness. In the tests described below, the fecal samples were used as supernatants ' of $0 'to PO Olio suspensions in PBS-D (see below) after centrifugation a t 16,300 x g. and 4 OC. If necessary, the supernatants were kept at -70 OC until tested. The identity of the virus present in feces was also verified by ID using an HA-positive sample, the reference CPV strain and the reference antiserum (Fig. 2) . In the original method of double diffusion only one precipitation line was formed (Fig. 2) . In WADSWORTH'S micromethod there were one very faint and two strong lines of identity (not shown), announcing the presence of three antigens. Three polypeptides are known to belong to the protein of some parvoviruses, e. g. the virus of feline panleukopenia (9). For routine diagnosis of CPV in the feces, the ID-test and counterimmunoelectrophoresis were found to be insensitive, i. e., only five of a total of 54 samples were positive in both these tests (data not shown). In one of the positive fecal samples we also found, by EM, reovirus-like particles. In three further sam les, one from a healthy bitch and two from its two diseased puppies, so-callecf "fringed" particles were seen in the absence of parvovirus-like particles. The "fringed" particles were similar to those observed in feces of calves with diarrhea (10, 11). Enterovirus-like particles were also seen in some samples; they were larger than parvovirus particles. Finally, Campylobacter spp were cultivated from about 60 O/o of fecal samples examined from hospital B (SANDSTEDT and HURVELL, personal communication).

CARMICHAEL'S procedure of detecting, in fecal samples, the CPV via its hemagglutinin (6) roved to be a rapid and reliable method; the specificity of H A could easily ; e verified by HI with a reference antiserum. However, using this procedure the percentage of positive dogs was low when compared with that detected by EM, i. e., 25.9 versus 48.3 O/o. On the other hand, in several samples the number of viral particles might have been too small to give HA or the particles to be identified as CPV by immunoaggregation. The presence of another parvovirus of dogs, the MCV (3) instead of CPV, is conceivable. Besides, in many routine laboratories an electron microscope is not available. The procedure of cultivating the CPV from feces in cell cultures proved to be time-consuming and unreliable (data not shown).

The infection of dogs with CPV is followed by development of an antibody response reaching comparatively high levels during the acute stage of illness. This could easily be demonstrated in HI-tests against a reference CPV strain.

For the HA-and HI-tests the erythrocytes from piglets were found to be much more suitable than those from older pigs whose erythrocytes tend to agglutinate spontaneously. The reason for this is unknown to us, but a changed charge in the membrane of erythrocytes from older pigs might be responsible.

It is not known to what extent the severity of enteritis was influenced by concomitant infections with Campylobacter spp. These bacteria are known to occur in do s which can be the source of infection and enteritis in man (5).

their possible role as disease agents is unknown.

Exceptional B y reovirus-like and "fringed" particles were seen in the feces;

Hemagglutination (HA) test and electron microscopy (EM) were used to diagnose canine parvoviral enteritis on fecal samples from 58 hospitalized dogs of a huge epidemic. By HA-tests the presence of the canine parvovirus (CPV) involved was shown in samples from 15 dogs (-26 O/o). A reference antiserum was used to identify the virus of each sample by hemagglutination inhibition (HI). By EM parvovirus-like particles were seen in samples from 28 dogs ( m 48 O/O). Particles in six samples were identified as CPV by immunoaggregation. Sera from 17 out of 19 dogs examined showed specific HI titers during the acute stage of illness. A high incidence of concomitant infection with Campylobacter spp was found by the bacteriologists. Thanks are due to 2. DINTER for helpful suggestions, M. SODERBERG for excellent technical assistance, and M. WIERUP for collectting samples. This investigation was supported by a grant from the Albert Hjarre Fond.

Wahrend einer grossen Epidemie von Parvovirus-Enteritis der Hunde (in Schweden) wurde der Nachweis von Virus in Kotproben mit Hilfe der Hamagglutination (HA) und Elektronenmikroskopie (EM) versucht; die Kotproben stammten von 58 hospitalisierten Hunden. Mit dem HA-Test liei3 sidi das ursachliche Parvovirus in den Proben von 15 Hunden nachweisen 

Mediante pruebas de hemaglutinacibn y microscopia electrbnica se diagnosticb parvovirus-enteritis en caninos en muestras fecales de 58 caninos hospitalizados a raiz de una extensa epidemia. Con la prueba de hemaglutinacibn se detect6 la presencia de virus en muestras fecales de 15 caninos (-26 O / o ) . Un antisuero de referencia se utilizb para la identificacibn del virus (prueba de inhibicibn de la hemaglutinacibn). Con microscopia electrbnica se identificb parvovirus en muestras fecales de 28 caninos (& 48 O/o). El virus fuk identificado en seis muestras fecales utilizando la prueba de inmunoagregacibn. Sueros de 17 de 19 caninos examinados demostraron titulos especificos de inhibicibn de la hemaglutinacibn durante la etapa aguda de la enfermedad. Se demostrb tambikn una alta incidencia de infeccibn concomitante con Cumpylobucter spp.

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