Molecular and serological detection of Theileria equi, Babesia caballi and Anaplasma phagocytophilum in horses and ticks in Maranhão, Brazil

Nogueira, Rita de Maria Seabra; Silva, Arannadia Barbosa; Sato, Tayra Pereira; Sá, Joicy Cortez de; Santos, Ana Clara Gomes dos; Amorim, Edvaldo Franco; Vale, Tássia Lopes do; Gazêta, Gilberto Salles

doi:10.1590/S0100-736X2017001200010

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LIVESTOCK DISEASES • Pesq. Vet. Bras. 37 (12) • Dec 2017 • https://doi.org/10.1590/S0100-736X2017001200010 copy

Molecular and serological detection of Theileria equi, Babesia caballi and Anaplasma phagocytophilum in horses and ticks in Maranhão, Brazil

Detecção molecular e serológica de Theileria equi, Babesia caballi e Anaplasma phagocytophilum em equinos e carrapatos no Maranhão

Authorship SCIMAGO INSTITUTIONS RANKINGS

ABSTRACT:

Equine piroplasmosis is a tick-borne disease caused by the intraeytrhocytic protozoans Babesia caballi and Theileria equi. It has been reported as a main equine parasitic disease. In addition, Anaplasma phagocytophilum, the causative agent of granulocytic ehrlichiosis, causes a seasonal disease in horses. Both diseases, can be detrimental to animal health. In this sense, blood samples and ticks were collected from 97 horses raised in the microregion of Baixada Maranhense, Maranhão State, Brazil. Serum samples were subjected to Indirect Fluorescence Antibody Test (IFAT) and blood samples and ticks to Polymerase Chain Reaction (PCR) to evaluate the infection by Theileria equi, Babesia caballi and Anaplasma phagocytophilum. The overall seroprevalence was 38.14%, 18.55% and 11.34% for T. equi, B. caballi and A. phagocytophilum, respectively. The results of PCR from blood samples showed 13.40% and 3.09% positive samples to T. equi and B. caballi, respectively. A total of 170 tick specimens were collected and identified as Dermacentor nitens, Amblyomma cajennense sensu lato and Rhipicephalus (Boophilus) microplus. It was detected 2.35% (4/170) and 0.59% (1/170) positive tick samples by PCR for T. equi and B. caballi, respectively. All samples were negative to A. phagocytophilum. No statically difference (p>0.05) was observed when gender, age, use of ectoparasiticide and tick presence were analyzed. A BLASTn analysis of the sequenced samples indicated 97 to 100% similarity with T. equi 18S rRNA gene sequences in GenBank and 98 to 100% with B. caballi. Genetic analysis classified the obtained sequences as T. equi and B. caballi cluster, respectively. It can be concluded that these pathogens occur and are circulating in the studied area.

INDEX TERMS:
Vector-borne disease; Theileria equi; Babesia caballi; Anaplasma phagocytophilum; horses; ticks; Brazil

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[1] *Corresponding author: grita62@hotmail.com

	Titers of IgG antibodies					Seroreactive animals
	1:80	1:160	1:320	1:640	1:1280	Seroreactive animals
Anti-T. equi	9	4	4	10	10	37
Anti-B. caballi	4	2	2	8	2	18
Anti- A. phagocytophilum	2	6	3	-	-	11

Variable		Reactive T. equi		Non reactive T. equi		Total	OR	IC	Value-P
Variable		N	%	N	%	Total	OR	IC	Value-P
Gender	M	9	26.47	25	73.53	34	0.45	0.16-1.22	0.1285628(a)
Gender	F	28	44.44	35	55.56	63	0.45	0.16-1.22	0.1285628(a)
Age (years)	<1	2	15.38	11	84.62	13
	1-4	5	18.52	22	81.48	27	0.80	0.09-6.00	0.5917915(b)
	5-8	17	53.12	15	46.88	32	0.16	0.02-0.98	0.0465580(a)*
	9- 2	10	62.50	6	37.50	16	0.11	0.01-0.84	0.0290414(a)*
	>13	3	33.33	6	66.67	9	0.36	0.03-3.96	0.3157895(b
Use of ectoparasiticide	Yes	20	37.74	33	62.26	53	0.96	0.39-2.38	0.9052469(a)
Use of ectoparasiticide	No	17	38.64	27	61.36	44	0.96	0.39-2.38	0.9052469(a)
Presence of ticks	Yes	32	37.65	53	63.35	85	0.85	0.22-3.40	0.5116261(a)
Presence of ticks	No	5	41.67	7	58.33	12	0.85	0.22-3.40	0.5116261(a)

Variable		Reactive B. caballi		Non reactive B. caballi		Total	OR	IC	Value-P
Variable		N	%	N	%	Total	OR	IC	Value-P
Gender	M	4	11.76	30	88.24	34	0.47	0.12-1.72	0.3219866(a)
Gender	F	14	22.22	49	77.78	63	0.47	0.12-1.72	0.3219866(a)
Age (years)	<1	-	-	-	-	13
	1-4	-	-	-	-	27
	5-8	11	34.38	21	65.62	32
	9-12	4	25	12	75	16	1.57	0.35-7.50	0.7411815(a)
	>13	3	33.34	6	66.66	9	1.05	0.18-6.63	0.6403036(b)
Use of ectoparasiticide	Yes	13	24.53	40	75.47	53	2.53	0.74-9.09	0.1620887(a)
Use of ectoparasiticide	No	5	11.36	39	88.64	44	2.53	0.74-9.09	0.1620887(a)
Presence of ticks	Yes	16	18.82	69	81.18	85	1.16	0.20-8.51	0.6092381(b)
Presence of ticks	No	2	16.67	10	83.33	12	1.16	0.20-8.51	0.6092381(b)

Variable		Reactive A. phagocytophilum		Non reactive A. phagocytophilum		Total	OR	IC	Value-P
Variable		N	%	N	%	Total	OR	IC	Value-P
Gender	M	4	11.76	30	88.24	34	1.07	0.24-4.53	0.5836453(a)
Gender	F	7	11.11	56	88.89	63	1.07	0.24-4.53	0.5836453(a)
Age (years)	< 1	3	23.07	10	76.93	13
	1-4	2	7.40	25	92.6	27	3.75	0.41-39.15	0.1838549(a)
	5 a 8	3	9.37	29	90.63	32	2.90	0.38-22.67	0.2229695(a)
	9 a 12	2	12.50	14	87.50	16	2.10	0.22-22.83	0.3961686(a)
	>13	1	11.11	8	88.89	9	2.40	0.16-73.20	0.4496241(a)
Use of ectoparasiticide	Yes	6	11.32	47	88.68	53	1.00	0.24-4.14	0.6200802(a)
Use of ectoparasiticide	No	5	11.36	39	88.64	44	1.00	0.24-4.14	0.6200802(a)
Presence of ticks	Yes	10	8.5	75	91.5	85	1.47	0.16-33.56	0.5919983(a)
Presence of ticks	No	1	8.33	11	91.67	12	1.47	0.16-33.56	0.5919983(a)