Enterites severas em cães associadas a infecções simples e mistas

Mortari, Ana Paula Gnocato; Seeger, Marlane Geribone; Oliveira, Pablo Sebastian Britto de; Masuda, Eduardo Kenji; Flores, Mariana Martins; Flores, Eduardo Furtado; Cargnelutti, Juliana Felipetto; Vogel, Fernanda Silveira Flôres

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Microbiology • Cienc. Rural 54 (2) • 2024 • https://doi.org/10.1590/0103-8478cr20220472 copiar

Enterites severas em cães associadas a infecções simples e mistas

Autoria SCIMAGO INSTITUTIONS RANKINGS

RESUMO:

Enterites infecciosas são altamente prevalentes entre cães em todo o mundo e, em alguns casos, podem ser fatais. Este estudo descreve os achados clínicos e laboratoriais de infecções simples e mistas associadas a enterites graves em 76 cães do sul do Brasil. Segmentos intestinais e/ou fezes foram submetidos à análise patológica e detecção molecular de vírus de genoma DNA, bactérias e protozoários. Lesões intestinais severas foram observadas na maioria dos casos. Infecções únicas foram detectadas em 52,6% dos casos, e infecções duplas (36,8%) e triplas (10,5%) também foram identificadas. O Protoparvovírus carnívoro 2 (CPV-2) foi o agente mais frequente nas infecções simples (36,8% do total). Coinfecção por CPV-2 e Giardia sp. foi a mais comum em infecções duplas (19,7%), seguido por CPV-2 e Cryptosporidium spp. (10,5%). A tripla infecção mais frequente foi CPV-2, Giardia spp. e Cryptosporidium spp. (6,6%). Nossos resultados demostraram que infecções simples e mistas estão associadas a enterites graves em cães no sul do Brasil, envolvendo principalmente CPV-2 e Giardia sp.

Palavras-chave:
gastroenterite; cães; bactérias; vírus; protozoários; diagnóstico

Autoria

Ana Paula Gnocato Mortari

Programa de Pós-graduação em Medicina Veterinária, Universidade Federal de Santa Maria (UFSM), Santa Maria, RS, Brasil.Universidade Federal de Santa Maria (UFSM)BrazilSanta Maria, RS, BrazilPrograma de Pós-graduação em Medicina Veterinária, Universidade Federal de Santa Maria (UFSM), Santa Maria, RS, Brasil.

https://orcid.org/0000-0002-0029-1296

Marlane Geribone Seeger

https://orcid.org/0000-0002-1226-2089

Pablo Sebastian Britto de Oliveira

https://orcid.org/0000-0002-9280-1766

Eduardo Kenji Masuda

Laboratório Axys Análises, Porto Alegre, RS, Brasil.Laboratório Axys AnálisesBrasilPorto Alegre, RS, BrasilLaboratório Axys Análises, Porto Alegre, RS, Brasil.

https://orcid.org/0000-0003-4093-4935

Mariana Martins Flores

Departamento de Patologia, Universidade Federal de Santa Maria (UFSM), Santa Maria, RS, Brasil.Universidade Federal de Santa Maria (UFSM)BrazilSanta Maria, RS, BrazilDepartamento de Patologia, Universidade Federal de Santa Maria (UFSM), Santa Maria, RS, Brasil.

https://orcid.org/0000-0002-2754-6885

Eduardo Furtado Flores

Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria (UFSM), 97105-900, Santa Maria, RS, Brasil..Universidade Federal de Santa Maria (UFSM)BrazilSanta Maria, RS, BrazilDepartamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria (UFSM), 97105-900, Santa Maria, RS, Brasil..

https://orcid.org/0000-0002-5074-6025

Juliana Felipetto Cargnelutti

https://orcid.org/0000-0002-3160-3643

Fernanda Silveira Flôres Vogel ^*E-mail: fernanda.vogel@ufsm.br. *Corresponding author

https://orcid.org/0000-0003-1351-9912

E-mail: fernanda.vogel@ufsm.br. ^*Corresponding author

Editor: Rudi Weiblen (0000-0002-1737-9817)

DECLARATION OF CONFLICT OF INTEREST

The authors declare no conflict of interest.

AUTHORS’ CONTRIBUTIONS

All authors contributed to perform research, interpreting the results, writing and critically revising the manuscript. All authors approved the final version of the manuscript.

SCIMAGO INSTITUTIONS RANKINGS

Laboratório Axys Análises, Porto Alegre, RS, Brasil.Laboratório Axys AnálisesBrasilPorto Alegre, RS, BrasilLaboratório Axys Análises, Porto Alegre, RS, Brasil.

Figuras | Tabelas

Figuras (1)
Tabelas (2)

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Figure 1
Pathological lesions observed in the intestine of dogs with severe enteritis caused by viruses, protozoa or bacteria. A) Necrosis and collapse of crypts, with villous atrophy and squamous metaplasia of intestinal cells in the small intestine of a dog infected with canine parvovirus type 2. Hematoxylin and eosin (HE). Bar: 150µm. B) Multifocal crypt abscesses in the small intestine of a dog infected with Giardia sp. Multiple dilated crypts are filled with cellular debris and degenerated neutrophils. HE, bar: 150 µm. C) Intestine of a dog infected with Clostridium perfringens A presenting massive necro-hemorrhagic enteritis. D) Myriad of basophilic bacilli lining a necrotic intestinal villus of a dog infected with Clostridium perfringens A. HE, bar: 150 µm.

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Table 1
Agent, target gene, primer sequences and expected amplicon size of PCR and nested-PCR used to detect DNA virus, bacteria and protozoa agents in feces and intestinal fragments of dogs with severe enteritis.

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Table 2
Infections and co-infections detected in dogs presenting severe enteritis in southern Brazil.

Figure 1 Pathological lesions observed in the intestine of dogs with severe enteritis caused by viruses, protozoa or bacteria. A) Necrosis and collapse of crypts, with villous atrophy and squamous metaplasia of intestinal cells in the small intestine of a dog infected with canine parvovirus type 2. Hematoxylin and eosin (HE). Bar: 150µm. B) Multifocal crypt abscesses in the small intestine of a dog infected with Giardia sp. Multiple dilated crypts are filled with cellular debris and degenerated neutrophils. HE, bar: 150 µm. C) Intestine of a dog infected with Clostridium perfringens A presenting massive necro-hemorrhagic enteritis. D) Myriad of basophilic bacilli lining a necrotic intestinal villus of a dog infected with Clostridium perfringens A. HE, bar: 150 µm.

Table 1 Agent, target gene, primer sequences and expected amplicon size of PCR and nested-PCR used to detect DNA virus, bacteria and protozoa agents in feces and intestinal fragments of dogs with severe enteritis.

Agent	Gene	Primers (5'-3')	Product (bp)	Reference
Giardia spp.	β-giardin (bg)	G7: AAGCCCGACGACCTCACCCGCAGTGC G759:GAGGCCGCCCTGGATCTTCGAGACGAC BG-Nst-F: GAACGAACGAGATCGAGGTCCG BG- Nst-R: CTCGACGAGCTTCGTGTT	753^a 511^b	(OSMARI et al., 2021OSMARI, V. et al. Occurrence and molecular characterization of Giardia duodenalis from naturally infected dogs in the municipality of Santa Maria, Rio Grande do Sul, Brazil. Pesquisa Veterinária Brasileira, v.41, e06670, 2021. Available from: <Available from: https://doi.org/10.1590/1678-5150-PVB-6670 >. Accessed: Jun. 17, 2021. doi: 10.1590/1678-5150-PVB-6670. https://doi.org/10.1590/1678-5150-PVB-66... )
Cryptosporidium spp.	SSU rRNA	F1: TTCTAGAGCTAATACATGCG R1: CCCATTTCCTTCGAAACAGGA F2: GGAAGGGTTGTATTTATTAGATAAAG R2: AAGGAGTAAGGAACAACCTCCA	1325^a 819-825^b	(ALVES et al., 2018ALVES, M. E. M. et al. Molecular detection of Cryptosporidium spp. and the occurrence of intestinal parasites in fecal samples of naturally infected dogs and cats. Parasitology Research, v.117, n.9, p.3033-3038, 2018. Available from: <Available from: https://doi.org/10.1007/s00436-018-5986-4 >. Accessed: Mar. 09, 2021. doi: 10.1007/s00436-018-5986-4. https://doi.org/10.1007/s00436-018-5986-... )
Sarcocystis spp.	18s rDNA	F: CGCAAATTACCCAATCCTGA R: ATTTCTCATAAGGTGCAGGAG	700	(PORTELLA et al., 2016PORTELLA, L. P. et al. Antibodies against Neospora caninum, Sarcocystis spp. and Toxoplasma gondii detected in buffaloes from Rio Grande do Sul, Brazil. Pesquisa Veterinaria Brasileira, v.46, n.9, p.1613-1617, 2016. Available from: <Available from: https://doi.org/10.1590/0103-8478cr20151365 >. Accessed: Jul. 29, 2022. doi: 10.1590/0103-8478cr20151365. https://doi.org/10.1590/0103-8478cr20151... )
Toxoplasma gondii	B1	F:CGCTGCAGGGAGGAAGACGAAAGTTG R:CGCTGCAGACACAGTGCATCTGGATT	529	(PORTELLA et al., 2016PORTELLA, L. P. et al. Antibodies against Neospora caninum, Sarcocystis spp. and Toxoplasma gondii detected in buffaloes from Rio Grande do Sul, Brazil. Pesquisa Veterinaria Brasileira, v.46, n.9, p.1613-1617, 2016. Available from: <Available from: https://doi.org/10.1590/0103-8478cr20151365 >. Accessed: Jul. 29, 2022. doi: 10.1590/0103-8478cr20151365. https://doi.org/10.1590/0103-8478cr20151... )
Neospora caninum	Nc5	F:CCCAGTGCGTCCAATCCTGTAAC R:CTCGCCAGTCAACCTACGTCTTCT	338	(PORTELLA et al., 2016PORTELLA, L. P. et al. Antibodies against Neospora caninum, Sarcocystis spp. and Toxoplasma gondii detected in buffaloes from Rio Grande do Sul, Brazil. Pesquisa Veterinaria Brasileira, v.46, n.9, p.1613-1617, 2016. Available from: <Available from: https://doi.org/10.1590/0103-8478cr20151365 >. Accessed: Jul. 29, 2022. doi: 10.1590/0103-8478cr20151365. https://doi.org/10.1590/0103-8478cr20151... )
Clostridium perfringens	Toxin A	CPA 5L: AGTCTACGCTTGGGATGGAA CPA 5R: TTTCCTGGGTTGTCCATTTC	900	(BURET, A. et al., 1992BURET, A. et al. Pathophysiology of small intestinal malabsorption in gerbils infected with Giardia lamblia. Gastroenterology, v.103, n.2, p.506-513, 1992. Available from: <Available from: http://linkinghub.elsevier.com/retrieve/pii/001650859290840U >. Accessed: Jun. 8, 2021. doi: 10.1016/0016-5085(92)90840-U. http://linkinghub.elsevier.com/retrieve/... )
Clostridium difficile	tcdA and tdB	NK2:CCCAATAGAAGATTCAATATTAAGCTT NK3: GGAAGAAAAGAACTTCTGGCTCACTCAGGT cdBFor: TAATAGAAAACAGTTAGAAA cdBRev: TCCAATCCAAACAAAATGTA	252^c 301^d	(ALVAREZ-PEREZ et al., 2009ALVAREZ-PEREZ, S. et al. Prevalence of Clostridium difficile in diarrhoeic and non-diarrhoeic piglets. Veterinary Microbiology, v.137, n.3-4, p.302-305, 2009. Available from: <Available from: https://doi.org/10.1016/j.vetmic.2009.01.015 >. Accessed: Jun. 08, 2021. doi: 10.1016/j.vetmic.2009.01.015. https://doi.org/10.1016/j.vetmic.2009.01... )
CanineCV	Rep	For_genomic:ATGGCTCAAGCTCAGGTTG Rev 533: CCGCACAGAACCTCCACTTC	533	(KOTSIAS et al., 2019KOTSIAS, F. et al. Genomic characterization of canine circovirus associated with fatal disease in dogs in South America. PloS one, v.14, n.6, p.e0218735, 2019. Available from: < Available from: https://pmc/articles/PMC6592543/ >. Accessed: Mar. 01, 2021. doi: 10.1371/journal.pone.0218735. https://pmc/articles/PMC6592543/... )
CPV-2	VP2	555_for:CAGGAAGATATCCAGAAGGA 555_rev: GGTGCTAGTTGATATGTAATAAACA	583	(BUONAVOGLIA, et al., 2001BUONAVOGLIA, C. et al. Evidence for evolution of canine parvovirus type 2 in Italy. Journal of General Virology, v.82, n.12, p.3021-3025, 2001. Available from: <Available from: https://doi.org/10.1099/0022-1317-82-12-3021 >. Accessed: Sep. 24, 2019. doi: 10.1099/0022-1317-82-12-3021. https://doi.org/10.1099/0022-1317-82-12-... )

a: primary reaction; b: secondary reaction; c: toxin A; d: toxin B.

Table 2 Infections and co-infections detected in dogs presenting severe enteritis in southern Brazil.

Type of infection	Number of cases	%
Single infection	40	52.6
CPV-2	28	36.8
Giardia sp.	9	11.8
Crysptosporidium sp.	2	2.6
Clostridium perfringens type A	1	1.3
Double infection	28	36.8
Giardia sp. and CPV-2	15	19.7
Cryptosporidium sp. and CPV-2	8	10.5
Giardia sp. and Cryptosporidium sp.	2	2.6
Giardia sp. and Clostridium perfringens type A	2	2.6
Clostridium perfringens type A and CPV-2	1	1.3
Triple infection	8	10.5
Giardia sp., Cryptosporidium sp. and CPV-2	5	6.6
Giardia sp., Sarcocystis sp. and CPV-2	1	1.3
Giardia sp., Clostridium difficile and CPV-2	1	1.3
Giardia sp., Clostridium perfringens type A and CPV-2	1	1.3
Total	76	100

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[1] E-mail: fernanda.vogel@ufsm.br. ^*Corresponding author