Molecular identification of Theileria species in naturally infected sheep using nested PCR–RFLP

• Published in: Parasitology research (1987) Vol. 121; no. 5; pp. 1487 - 1497
• Main Authors: Nangru, Aman, Maharana, B. R., Vohra, Sukhdeep, Kumar, Binod
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01-05-2022; Springer; Springer Nature B.V
• Subjects: Analysis; Animals; Biomedical and Life Sciences; Biomedicine; Cattle; Enzymes; Ethylenediaminetetraacetic acid; Health aspects; Immunology; Infection; Medical Microbiology; Microbiology; Mixed infection; Polymerase chain reaction; Polymerase Chain Reaction - veterinary; Polymorphism, Restriction Fragment Length; Protozoology - Original Paper; Restriction fragment length polymorphism; Sequence analysis; Sheep; Sheep Diseases; Theileria; Theileria - genetics; Theileriasis - epidemiology; Theileriosis; Ticks; Iran; Lithuania; China; India; PCR–RFLP; Ovine theileriosis; 18S rRNA; Molecular identification; India
• ISSN: 0932-0113; 1432-1955
• DOI: 10.1007/s00436-022-07489-5

Ovine theileriosis is an important tick-borne haemoprotozoan disease of sheep in tropical and subtropical regions, causing severe productivity and economic loss. There is a paucity of information related to molecular studies of ovine theileriosis from India. The present study identified different
Theileria
spp. in naturally infected sheep using nested PCR-restriction fragment length polymorphism (nPCR-RFLP). Blood samples and ticks were collected from 204 sheep in different agro-climatic zones of Haryana state, India, during the tick active season. Microscopic examination of thin blood smears revealed 33.3% (68/204) infections with
Theileria
spp., while 44.6% (91/204) of blood samples were positive by nPCR assay. Different
Theileria
spp
.
were identified based upon RFLP patterns using four restriction enzymes:
Hpa
II,
Bsh
1285I,
Hae
II and
Rsa
I. Out of 91 positive samples, 50.5% (46/91), 23.08% (21/91), 11% (10/91) and 2.2% (2/91) were positive for
T. ovis
,
T. lestoquardi
,
T. luwenshuni
(
Theileria
sp. China 1/
Theileria
sp. China) and
T. annulata
, respectively
.
Mixed infection was detected in 13.2% (12/91) of cases. Based upon
Hpa
II enzymatic digestion pattern, two samples with
T. lestoquardi
and
T. annulata
, nine samples with
T. lestoquardi
and
T. ovis
and one sample with
T. ovis
and
T. annulata
were detected. The presence of these
Theileria
spp. was further confirmed by sequence analysis. The majority of ticks collected from sheep were identified as
Rhipicephalus
spp
.
followed by
Hyalomma anatolicum
and
Hemaphysalis
spp. The present investigation depicts the first comprehensive molecular report of naturally infected sheep with
T. ovis
,
T. lestoquardi
,
T. annulata
and
T. luwenshuni
from northern India.