Identificação morfológica e molecular com análise de frequência de <i>Calicophoron microbothrioides</i> na região central do Chile

Cerda, Carolina; Veloso-Frías, Joseline; Lobos-Chávez, Felipe; Oyarzún-Ruiz, Pablo; Henríquez, AnaLía; Loyola, Marco; Fuente, María Carolina Silva-de la; Ortega, René; Letelier, Reinaldo; Landaeta-Aqueveque, Carlos

doi:10.1590/S1984-29612019076

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Revista Brasileira de Parasitologia Veterinária

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Original Article • Rev. Bras. Parasitol. Vet. 28 (4) • Oct-Dec 2019 • https://doi.org/10.1590/S1984-29612019076 copiar

Identificação morfológica e molecular com análise de frequência de Calicophoron microbothrioides na região central do Chile

Autoria SCIMAGO INSTITUTIONS RANKINGS

Resumo

Este trabalho teve como objetivo determinar a presença de paramphistomídeos em bovinos abatidos em um matadouro da Região do Ñuble do Chile, para identificar parasitas e analisar a frequência desses parasitos nas regiões administrativas de Maule, Ñuble e Biobío, no Chile. Entre outubro de 2016 e abril de 2017, rúmens de 494 bovinos foram examinados à procura de vermes no pré-estômago. Os vermes foram identificados morfologicamente e, além disso, a análise molecular da região interna do espaçador do transcritor 2 do DNA e análises filogenéticas foram realizadas com inferência bayesiana em 14 vermes. A frequência foi analisada pela altitude da localidade (baixa ou alta) e idade. A frequência geral foi de 11,24%. O distrito com as maiores frequências de parasitismo foi Chillán Viejo (30,8%). Os distritos das terras baixas tinham frequências semelhantes às encontradas nas terras das montanhas (p=0,17). A frequência foi significativamente maior em animais adultos do que em jovens (p<0.01). Obtivemos um fragmento de DNA de 460 pb que era idêntico às sequências anteriores identificadas como Paramphistomum cervi e Calicophoron microbothrioides, e realizamos análises morfológicas que permitiram confirmar que nossas amostras pertenciam a C. microbothrioides. Este é o primeiro estudo publicado sobre C. microbothrioides no Chile.

Palavras-chave:
Paramphistomidae; vermes ruminais; bovino; epidemiologia; ITS-2; Calicophoron microbothrioides

Province	District/Code	Examined Animals	Frequency of presentation [%] (95%, confidence interval)	Mountain or flat lowlands district
Cauquenes	Cauquenes/1	1	100 (5 – 100)	Mountain
Linares	Linares/2	3	66.67 (13.32 – 100)	Mountain
Punilla	Ñiquén/3	21	28.57 (9.25 - 47.89)	Lowland
	San Carlos/4	178	11.80 (7.06 - 19.54)	Lowland
	San Nicolás/5	26	3.85 (0.0 - 11.24)	Lowland
	Coihueco/6	24	25 (7.68 - 42.32)	Mountain
Diguillín	Chillan/7	7	0 (0 - 34.8)	Lowland
	Chillan Viejo/8	13	30.77 (5.68 - 55.86)	Lowland
	Pinto/9	7	0 (0 - 34.8)	Mountain
	Bulnes/10	73	4.11 (0.0 - 8.66)	Lowland
	San Ignacio/11-El Carmen/12	10	0 (0 - 25.9)	Mountain
Itata	Quirihue/13	12	16.67 (9.25 - 47.89)	Mountain
	Ninhue/14-Coelemu/15	7	14.29 (0 - 40.21)	Mountain
	Portezuelo/16-Quillón/17	17	11.76 (0 - 27.08)	Lowland
Concepción	San Pedro de la Paz/18	2	0 (0 - 77.64)	Lowland
Arauco	Arauco/19	3	0 (0 - 63.16)	Lowland
Biobío	Cabrero/20	4	0 (0 - 52.71)	Lowland
	Los Ángeles/21	59	6.78 (0.36 - 13.19)	Lowland
	Mulchén/22-Santa Bárbara/23-Tucapel/24	27	11.11 (0 - 22.97)	Mountain
	Total	494	11.34 (8.54 – 14.13)

		1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21	22	23	24	25	26	27	28	29	30	31	32	33	34	35	36	37	38	39	40
1	AB042188-Calicophoron_calicophorum
2	AB042189-Orthocoelium_streptocoelium	0.034
3	AB056570-Calicophoron_microbothrioides	0.026	0.029
4	AB743577Explanatum_explanatum	0.035	0.026	0.026
5	AB973398Paramphistomum_leydini	0.038	0.040	0.035	0.020
6	AY790883Calicophoron_daubneyi	0.031	0.040	0.029	0.031	0.029
7	EF027097Gastrodiscoides_hominis	0.072	0.062	0.069	0.065	0.071	0.075
8	EU887295Gastrodiscoides_hominis	0.037	0.034	0.040	0.037	0.046	0.040	0.036
9	GU133057Calicophoron_calicophorum	0.006	0.037	0.020	0.029	0.032	0.026	0.072	0.037
10	GU133058Orthocoelium_streptocoelium	0.040	0.008	0.029	0.026	0.040	0.037	0.065	0.037	0.037
11	HM209064Paramphistomum_leydeni	0.035	0.037	0.032	0.017	0.003	0.026	0.071	0.043	0.029	0.037
12	HM209067Paramphistomum_leydeni	0.035	0.037	0.032	0.017	0.003	0.026	0.071	0.043	0.029	0.037	0.000
13	ID11Chile	0.026	0.029	0.000	0.026	0.035	0.029	0.069	0.040	0.020	0.029	0.032	0.032
14	ID12Chile	0.026	0.029	0.000	0.026	0.035	0.029	0.069	0.040	0.020	0.029	0.032	0.032	0.000
15	JF834888-Paramphistomum_epiclitum	0.032	0.038	0.044	0.035	0.044	0.043	0.081	0.046	0.035	0.038	0.041	0.041	0.044	0.044
16	JX678251-Calicophoron_shillongensis	0.020	0.029	0.023	0.020	0.023	0.023	0.069	0.034	0.014	0.029	0.020	0.020	0.023	0.023	0.026
17	JX678256-Leiperocotyle_meghalayensis	0.029	0.014	0.023	0.026	0.034	0.034	0.066	0.031	0.026	0.011	0.032	0.032	0.023	0.023	0.032	0.023
18	KF564869-Explanatum_explanatum	0.038	0.029	0.029	0.003	0.023	0.034	0.068	0.040	0.032	0.029	0.020	0.020	0.029	0.029	0.038	0.023	0.029
19	KF564870-Paramphistomum_epiclitum	0.032	0.037	0.044	0.035	0.044	0.043	0.081	0.046	0.035	0.038	0.041	0.041	0.044	0.044	0.006	0.026	0.032	0.038
20	KF791030-Calicophoron_microbothrioides	0.026	0.029	0.000	0.026	0.035	0.029	0.069	0.040	0.020	0.029	0.032	0.032	0.000	0.000	0.044	0.023	0.023	0.029	0.044
21	KJ459934Paramphistomum_cervi	0.026	0.029	0.000	0.026	0.035	0.029	0.069	0.040	0.020	0.029	0.032	0.032	0.000	0.000	0.044	0.023	0.023	0.029	0.044	0.000
22	KJ459935Paramphistomum_cervi	0.026	0.035	0.023	0.026	0.029	0.023	0.059	0.026	0.020	0.035	0.026	0.026	0.023	0.023	0.038	0.017	0.029	0.029	0.038	0.023	0.023
23	KJ459936Paramphistomum_cervi	0.026	0.035	0.023	0.026	0.029	0.023	0.059	0.026	0.020	0.035	0.026	0.026	0.023	0.023	0.038	0.017	0.029	0.029	0.038	0.023	0.023	0.000
24	KJ459937Paramphistomum_cervi	0.026	0.029	0.000	0.026	0.035	0.029	0.069	0.040	0.020	0.029	0.032	0.032	0.000	0.000	0.044	0.023	0.023	0.029	0.044	0.000	0.000	0.023	0.023
25	KJ459938Paramphistomum_cervi	0.026	0.029	0.000	0.026	0.035	0.029	0.069	0.040	0.020	0.029	0.032	0.032	0.000	0.000	0.044	0.023	0.023	0.029	0.044	0.000	0.000	0.023	0.023	0.000
26	KP201674Calicophoron_daubneyi	0.031	0.040	0.029	0.031	0.029	0.000	0.075	0.040	0.026	0.037	0.026	0.026	0.029	0.029	0.043	0.023	0.034	0.034	0.043	0.029	0.029	0.023	0.023	0.029	0.029
27	KP639631Calicophoron_microbothrium	0.020	0.029	0.017	0.020	0.017	0.017	0.069	0.034	0.014	0.029	0.014	0.014	0.017	0.017	0.032	0.011	0.017	0.023	0.032	0.017	0.017	0.017	0.017	0.017	0.017	0.017
28	KP639632Calicophoron_microbothrium	0.020	0.029	0.017	0.020	0.017	0.017	0.069	0.034	0.014	0.029	0.014	0.014	0.017	0.017	0.032	0.011	0.017	0.023	0.032	0.017	0.017	0.017	0.017	0.017	0.017	0.017	0.000
29	KP639637Calicophoron_microbothrium	0.023	0.026	0.026	0.023	0.020	0.020	0.063	0.029	0.023	0.029	0.017	0.017	0.026	0.026	0.032	0.014	0.023	0.026	0.032	0.026	0.026	0.020	0.020	0.026	0.026	0.020	0.008	0.008
30	KP639638Calicophoron_microbothrium	0.020	0.034	0.017	0.026	0.023	0.017	0.066	0.031	0.014	0.034	0.020	0.020	0.017	0.017	0.038	0.017	0.023	0.029	0.037	0.017	0.017	0.017	0.017	0.017	0.017	0.017	0.006	0.006	0.008
31	KX840344-Paramphistomum_epiclitum	0.032	0.038	0.044	0.035	0.044	0.043	0.081	0.046	0.035	0.038	0.041	0.041	0.044	0.044	0.006	0.026	0.032	0.038	0.006	0.044	0.044	0.038	0.038	0.044	0.044	0.043	0.032	0.032	0.032	0.038
32	KX840345-Paramphistomum_epiclitum	0.029	0.035	0.041	0.032	0.041	0.040	0.077	0.044	0.032	0.035	0.038	0.038	0.041	0.041	0.008	0.023	0.029	0.035	0.008	0.041	0.041	0.035	0.035	0.041	0.041	0.040	0.029	0.029	0.029	0.035	0.003
33	LC101682Explanatum_explanatum	0.035	0.026	0.026	0.000	0.020	0.031	0.065	0.037	0.029	0.026	0.017	0.017	0.026	0.026	0.035	0.020	0.026	0.003	0.035	0.026	0.026	0.026	0.026	0.026	0.026	0.031	0.020	0.020	0.023	0.026	0.035	0.032
34	LC101683-Explanatum_explanatum	0.038	0.026	0.029	0.003	0.023	0.034	0.068	0.040	0.032	0.026	0.020	0.020	0.029	0.029	0.038	0.023	0.029	0.006	0.038	0.029	0.029	0.029	0.029	0.029	0.029	0.034	0.023	0.023	0.026	0.029	0.038	0.035	0.003
35	MK156397-Explanatum_explanatum	0.035	0.026	0.026	0.000	0.020	0.031	0.065	0.037	0.029	0.026	0.017	0.017	0.026	0.026	0.035	0.020	0.026	0.003	0.035	0.026	0.026	0.026	0.026	0.026	0.026	0.031	0.020	0.020	0.023	0.026	0.035	0.032	0.000	0.003
36	JX678257-Cotylophoron_cotylophorum	0.047	0.050	0.038	0.047	0.050	0.038	0.086	0.049	0.041	0.050	0.047	0.047	0.038	0.038	0.053	0.038	0.044	0.050	0.053	0.038	0.038	0.038	0.038	0.038	0.038	0.038	0.038	0.038	0.041	0.038	0.053	0.050	0.047	0.050	0.047
37	KF543340_Fasciola_gigantica	0.438	0.459	0.456	0.460	0.454	0.464	0.494	0.482	0.451	0.458	0.449	0.449	0.456	0.456	0.467	0.469	0.472	0.454	0.466	0.456	0.456	0.468	0.468	0.456	0.456	0.464	0.455	0.455	0.455	0.461	0.467	0.467	0.460	0.459	0.460	0.480
38	MG569981_Fasciola_hepatica	0.450	0.471	0.456	0.460	0.455	0.464	0.507	0.494	0.450	0.464	0.449	0.449	0.456	0.456	0.461	0.468	0.478	0.454	0.460	0.456	0.456	0.468	0.468	0.456	0.456	0.464	0.455	0.455	0.467	0.461	0.461	0.461	0.460	0.459	0.460	0.480	0.014
39	MG569979_Fasciola_hepatica	0.450	0.470	0.456	0.460	0.454	0.463	0.507	0.494	0.450	0.464	0.449	0.449	0.456	0.456	0.461	0.468	0.478	0.454	0.460	0.456	0.456	0.468	0.468	0.456	0.456	0.463	0.455	0.455	0.467	0.461	0.461	0.461	0.460	0.459	0.460	0.480	0.011	0.003
40	HQ197358_Fasciola_gigantica	0.445	0.465	0.463	0.467	0.461	0.470	0.501	0.488	0.457	0.465	0.455	0.455	0.463	0.463	0.461	0.475	0.478	0.461	0.460	0.463	0.463	0.475	0.475	0.463	0.463	0.470	0.461	0.461	0.461	0.467	0.461	0.461	0.467	0.466	0.467	0.486	0.003	0.011	0.008
41	KF791030.1_Calicophoron_microbothrioides	0.026	0.029	0.000	0.026	0.035	0.029	0.069	0.040	0.020	0.029	0.032	0.032	0.000	0.000	0.044	0.023	0.023	0.029	0.044	0.000	0.000	0.023	0.023	0.000	0.000	0.029	0.017	0.017	0.026	0.017	0.044	0.041	0.026	0.029	0.026	0.038	0.456	0.456	0.456	0.463

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[1] *Corresponding author: Carlos Landaeta-Aqueveque. Facultad de Ciencias Veterinarias, Universidad de Concepción, Vicente Méndez, 595, Chillán, Chile. e-mail: clandaeta@udec.cl