Canine visceral leishmaniasis in area with recent <i>Leishmania</i> transmission: prevalence, diagnosis, and molecular identification of the infecting species

Lopes, Josiane Valadão; Michalsky, Érika Monteiro; Pereira, Nathália Cristina Lima; Paula, Adão Junior Viana de; Souza, Andreza Geisiane Maia; Pinheiro, Letícia Cavalari; Lima, Ana Cristina Vianna Mariano da Rocha; Avelar, Daniel Moreira de; França-Silva, João Carlos; Lanzetta, Virgínia Aguiar Sorice; Melo, Jarbas de; Fortes-Dias, Consuelo Latorre; Dias, Edelberto Santos

doi:10.1590/0037-8682-0141-2020

Josiane Valadão Lopes

Instituto René Rachou, Fundação Oswaldo Cruz Minas, Belo Horizonte, MG, Brasil.

Érika Monteiro Michalsky

Instituto René Rachou, Fundação Oswaldo Cruz Minas, Belo Horizonte, MG, Brasil.

Nathália Cristina Lima Pereira

Instituto René Rachou, Fundação Oswaldo Cruz Minas, Belo Horizonte, MG, Brasil.

Adão Junior Viana de Paula

Instituto René Rachou, Fundação Oswaldo Cruz Minas, Belo Horizonte, MG, Brasil.

Andreza Geisiane Maia Souza

Instituto René Rachou, Fundação Oswaldo Cruz Minas, Belo Horizonte, MG, Brasil.

Letícia Cavalari Pinheiro

Instituto René Rachou, Fundação Oswaldo Cruz Minas, Belo Horizonte, MG, Brasil.

Ana Cristina Vianna Mariano da Rocha Lima

Instituto René Rachou, Fundação Oswaldo Cruz Minas, Belo Horizonte, MG, Brasil.

Daniel Moreira de Avelar

Instituto René Rachou, Fundação Oswaldo Cruz Minas, Belo Horizonte, MG, Brasil.

João Carlos França-Silva

Universidade Federal de Minas Gerais, Departamento de Parasitologia, Belo Horizonte, MG, Brasil.

Virgínia Aguiar Sorice Lanzetta

Prefeitura Municipal de Itaúna, Centro de Controle de Zoonoses, Itaúna, MG, Brasil.

Jarbas de Melo

Prefeitura Municipal de Itaúna, Centro de Controle de Zoonoses, Itaúna, MG, Brasil.

Consuelo Latorre Fortes-Dias

Fundação Ezequiel Dias, Diretoria de Pesquisa e Desenvolvimento, Belo Horizonte, MG, Brasil.

Edelberto Santos Dias

Instituto René Rachou, Fundação Oswaldo Cruz Minas, Belo Horizonte, MG, Brasil.

https://orcid.org/0000-0002-2370-5113

Corresponding author: Dr. Edelberto Santos Dias. e-mail:edelberto.dias@fiocruz.br

Author’s Contributions:JVL, EMM, NCLP, AGMS e AJVP: field work and laboratory experiments; JCFS: necropsy and euthanasia of seropositive dogs; DMA and ACVMRL: parasitological diagnosis (smear/imprint and myeloculture);LCP and CLFD: figures and statistical analyses; VASL and JM: logistic support in field work; JVL, EMM, EDS and CLFD: prepared the manuscript. All authors have read and approved the final version of the article.

Conflict of Interest: The authors declare no conflicts of interest.

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FIGURE 1:
Leishmania detection in dogs seropositive for canine visceral leishmaniasis (CVL). (A) Positivity rates of parasitological and molecular tests. (B) Test positivity according to the clinical group. Statistically significant differences between the two groups were observed only for myelocultures (indicated by *) (p < 0.05). (C) Frequency of positive tests according to clinical group. Each horizontal bar represents one dog (n = 60). Itaúna, Minas Gerais state, Brazil, 2016.

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FIGURE 2:
Visceral leishmaniasis scenario in Itaúna (Minas Gerais state, Brazil), an area of sporadic epidemiological transmission risk (ETR) for the disease. Prevalence of canine visceral leishmaniasis (CVL) for each neighborhood is indicated by background color: < 50 (yellow), 50-99 (orange), ≥ 100 (red). A white background indicates non-studied districts. Human visceral leishmaniasis (VL) cases are indicated by green stars and year of report. Sites of L. longipalpis capture are represented by circles. Closed circles denote sites of L. longipalpis capture which were subsequently found to be infected by Leishmania¹¹. Neighborhoods: A. Centro, B. Chácara do Quitão, C. Cidade Nova, D. Cidade Leonane, E. Garcias, F. Graças, G. Morada Nova, H. Nogueirinha, I. Novo Horizonte, J. Olaria, K. Parque Jardim Santanense, L. Três Marias, M. Vila Nazaré.

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TABLE 1:
Diagnostic test results and calculated prevalence of canine visceral leishmaniasis by Itaúna city neighborhood in the Brazilian state of Minas Gerais, 2016. CCS: canine census survey, SOD: spontaneous owner’s demand. TPR: test positivity rate.

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TABLE 2:
General characteristics of the subsample of CVL-seropositive dogs (n = 60) in an area of recent transmission of visceral leishmaniases. Itaúna, Minas Gerais state, Brazil, 2016.

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TABLE 3:
Parasitological and molecular test results for Leishmania in CVL-seropositive dogs. Itaúna, Minas Gerais state, Brazil, 2016.

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Neighborhood	CCS				SOD				Total (CCS plus SOD)				CVL prevalence^a
	Dogs	Diagnosis test results		TPR	Dogs	Diagnosis test results		TPR	Dogs	Diagnosis test results		TPR
	tested (no.)	DPP+	DPP+ ELISA+	(%)	tested (no.)	DPP+	DPP+ELISA +	(%)	tested (no.)	DPP +	DPP+ ELISA+	(%)
Centro	254	60	21	8.3	61	18	16	26.2	315	78	37	11.7	21.4
Chácara do Quitão	44	10	8	18.2	79	26	19	24.1	123	36	27	22.0	100.0
Cidade Nova	343	64	39	11.4	92	52	26	28.3	435	116	65	14.9	116.8
Cidade Leonane	186	34	19	10.2	27	11	11	40.7	213	45	30	14.1	97.4
Garcias	71	6	5	7	32	28	19	59.4	103	34	24	23.3	13.9
Graças	166	43	23	13.9	25	14	10	40.0	191	57	33	17.3	62.7
Morada Nova	15	0	0	0	83	60	47	56.6	98	60	47	48.0	0.0
Nogueirinha	90	17	15	16.7	4	2	1	25.0	94	19	16	17.0	166.7
Novo Horizonte	143	45	15	10.5	0	0	0	-	143	45	15	10.5	115.4
Olaria	42	9	6	14.3	45	27	11	24.4	87	36	17	19.5	17.1
P. J. Santanense	163	22	16	9.8	116	37	22	19.0	279	59	38	13.6	29.3
Três Marias	154	39	30	19.5	21	8	7	33.3	175	47	37	21.1	104.5
Vila Nazaré	19	9	6	31.6	27	21	14	51.9	46	30	20	43.5	51.3
Total	1690	358	203	12.0	612	304	203	33.2	2302	662	406	17.6	-
Mean ± SD	-	-	-	13.2±7.5	-	-	-	35.7 ±13.8	-	-		21.3±11.6	69.0±51.4

Variable	Dogs
	No.	%
Clinical classification
Asymptomatic	22	36.6
Symptomatic	38	63.4
Gender
Female	34	56.6
Male	26	43.4
Breed
Defined¹	16	26.6
Undefined	44	73.4
Hair length
Short	44	73.4
Long	16	26.6
Size
Small	21	35.0
Medium	28	46.6
Big	11	18.4

Clinical status	Dog no.	Parasitological						Molecular
			Slide imprint/smear					Bone	Lymph	Spleen	Ski n	Any
		Myeloculture	Bone marrow	Lymph node	Spleen	Skin	Any tissue	marrow	node			tissue
Asymp	4	-	-	+	+	-	+	+	+	+	+	+
	8	-	-	+	+	+	+	+	+	+	+	+
	9	+	+	+	+	+	+	+	+	+	+	+
	10	+	-	-	+	+	+	+	+	+	+	+
	11	-	+	-	+	-	+	+	+	+	+	+
	12	+	+	+	+	+	+	+	+	+	+	+
	15	+	-	-	-	-	-	+	+	+	+	+
	19	+	-	-	-	-	-	+	+	+	+	+
	22	-	-	+	+	+	+	+	+	+	+	+
	23	-	-	-	-	-	-	+	+	+	+	+
	27	-	-	-	+	-	+	+	+	+	+	+
	28	+	-	+	+	+	+	+	+	+	+	+
	32	-	+	+	+	+	+	+	+	+	+	+
	34	+	+	+	+	+	+	+	+	+	+	+
	48	-	-	+	+	-	+	+	+	+	+	+
	49	-	-	+	+	+	+	+	+	+	-	+
	51	+	-	-	-	-	-	+	+	+	+	+
	52	+	+	+	+	+	+	+	+	+	+	+
	53	+	-	-	+	-	+	+	+	+	+	+
	55	+	-	-	-	-	-	+	+	+	+	+
	57	-	-	-	-	-	-	+	+	-	+	+
	60	+	+	+	+	+	+	+	+	+	+	+
Symp	1	+	-	-	-	-	-	+	+	+	+	+
	2	+	-	-	-	-	-	+	+	+	+	+
	3	+	-	+	-	+	+	+	+	+	+	+
	5	+	-	-	-	+	+	+	+	+	+	+
	6	+	+	+	+	+	+	+	+	+	+	+
	7	-	+	+	+	+	+	+	+	+	+	+
	13	+	-	-	-	-	-	+	+	+	+	+
	14	+	-	-	-	-	-	+	+	+	+	+
	16	+	+	+	+	+	+	+	+	+	+	+
	17	+	+	+	-	+	+	+	-	+	+	+
	18	+	+	+	+	-	+	+	+	+	+	+
	20	+	-	-	-	-	-	+	+	+	+	+
	21	+	+	+	+	+	+	+	+	+	-	+
	24	-	-	-	-	-	-	+	+	+	-	+
	25	+	+	+	+	-	+	n.d.	+	+	+	+
	26	+	-	+	+	+	+	+	+	+	+	+
	29	+	+	+	+	+	+	+	+	+	+	+
	30	+	+	+	+	+	+	+	+	+	+	+
	31	+	-	-	+	-	+	+	+	+	+	+
	33	+	+	+	+	-	+	+	+	+	+	+
	35	+	+	+	+	-	+	+	+	+	+	+
	36	+	+	+	-	+	+	+	+	+	+	+
	37	+	-	+	+	+	+	+	+	+	+	+
	38	+	+	+	+	+	+	+	+	+	+	+
	39	+	+	+	+	+	+	+	+	+	+	+
	40	+	+	+	+	+	+	+	+	+	+	+
	41	+	+	+	+	+	+	+	+	+	+	+
	42	-	-	-	-	-	-	+	+	+	+	+
	43	+	+	+	+	+	+	+	-	+	+	+
	44	+	+	+	+	+	+	+	+	+	+	+
	45	+	-	+	+	-	+	+	+	+	+	+
	46	+	-	-	+	-	+	+	+	+	+	+
	47	-	-	-	-	-	-	-	-	+	+	+
	50	+	+	+	+	+	+	+	+	+	+	+
	54	+	+	+	+	+	+	+	+	+	+	+
	56	-	-	-	-	-	-	+	+	+	+	+
	58	+	-	+	+	-	+	+	+	+	+	+
	59	+	-	+	+	-	+	+	-	+	+	+
Positive/total		45/60	27/60	38/60	41/60	31/60	45/60	58/59	56/60	59/60	57/60	60/60

[TFN1] ^(a)
Calculated based on CCS data only.

Canine visceral leishmaniasis in area with recent Leishmania transmission: prevalence, diagnosis, and molecular identification of the infecting species

Abstract