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Development of a pan-Babesia FRET-qPCR and a survey of livestock from five Caribbean islands.

Li J, Kelly P, Zhang J, Xu C, Wang C - BMC Vet. Res. (2015)

Bottom Line: Babesia spp. are tick-borne protozoan hemoparasites and the second most common blood-borne parasites of mammals, in particular domestic animals.We used the Clustal Multiple Alignment program and 18S rRNA gene sequences of 22 Babesia species from GenBank to develop a PCR that could detect a wide variety of Babesia spp. in a single reaction.The pan-Babesia FRET-qPCR we developed reliably detected B. gibsoni, B. canis, B. vogeli, B. microti, B. bovis, and B. divergens under controlled conditions but did not react with closely related species, mainly Hepatozoon americanum, Theileria equi, and Toxoplasma gondii.

View Article: PubMed Central - PubMed

Affiliation: Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University College of Animal Science and Technology, Yangzhou, Jiangsu, 225009, P. R. China. lijing900401@163.com.

ABSTRACT

Background: Babesia spp. are tick-borne protozoan hemoparasites and the second most common blood-borne parasites of mammals, in particular domestic animals. We used the Clustal Multiple Alignment program and 18S rRNA gene sequences of 22 Babesia species from GenBank to develop a PCR that could detect a wide variety of Babesia spp. in a single reaction. The pan-Babesia FRET-qPCR we developed reliably detected B. gibsoni, B. canis, B. vogeli, B. microti, B. bovis, and B. divergens under controlled conditions but did not react with closely related species, mainly Hepatozoon americanum, Theileria equi, and Toxoplasma gondii.

Results: When we tested the pan-Babesia FRET-qPCR on DNA of whole blood from 752 cattle, sheep, goats, donkeys and horses from five Caribbean islands, we detected Babesia spp. expected to be present in the animals, mainly B. bovis and B. bigemina in cattle and B. caballi in horses and donkeys. Further, we found that animals were not uncommonly infected with species of Babesia usually associated with other hosts, mainly B. vogeli and B. gibsoni in cattle, sheep and goats, B. rossi in goats, and B. caballi in goats and sheep. Finally, the pan-Babesia FRET-qPCR enabled us to identify unknown species of Babesia in cattle, goats, sheep and donkeys.

Conclusions: Overall, 70 % (525/752) of the animals we tested were positive confirming earlier limited studies that infections with Babesia spp. are common in livestock in the Caribbean.

No MeSH data available.


Related in: MedlinePlus

Alignment of the partial 18S rRNA gene amplicons of Babesia spp. and other related species. The upstream primer-1 (in red), upstream primer-2 (in blue), the fluorescein/LCRed 640 probes and the downstream primer are indicated in the top of the boxes. Dots indicate nucleotides identical to the primers and probes, and the dashes denote the deletion of the nucleotides. The upstream primers and two probes are used as the indicated sequences while the downstream primer is used as antisense oligonucleotide. While the probes and downstream primer show minimum mismatch with Babesia spp. and other related species, the upstream primers (−1 and −2) have 0–1 nucleotide mismatch with Babesia spp. but 6–16 nucleotide mismatches with the related non- Babesia species
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Fig1: Alignment of the partial 18S rRNA gene amplicons of Babesia spp. and other related species. The upstream primer-1 (in red), upstream primer-2 (in blue), the fluorescein/LCRed 640 probes and the downstream primer are indicated in the top of the boxes. Dots indicate nucleotides identical to the primers and probes, and the dashes denote the deletion of the nucleotides. The upstream primers and two probes are used as the indicated sequences while the downstream primer is used as antisense oligonucleotide. While the probes and downstream primer show minimum mismatch with Babesia spp. and other related species, the upstream primers (−1 and −2) have 0–1 nucleotide mismatch with Babesia spp. but 6–16 nucleotide mismatches with the related non- Babesia species

Mentions: The 18S rRNA sequences for 22 recognized Babesia spp. of public health significance and/or veterinary importance were obtained from GenBank (Fig. 1): B. microti (AB071177, AB219802), B. leo (AF244911, AY452708), B. rodhaini (DQ641423, AB049999), B. felis (AF244912, AY452707), B. poelea (DQ200887), B. bigemina (JQ723014, KF112076), B. bovis (HQ264124, HQ264127), B. caballi (AY534883), B. canis (AY072926, JN982353), B. capreoli (FJ944828, GQ304526), B. crassa (AY260176, JX542614), B. divergens (FJ944822, FJ944826), B. gibsoni (KJ142323), B. hongkongensis (JQ867356), B. kiwiensis (EF55315), B. major (JF802040), B. motasi (AY260179, AY533147), B. odocoilei (AY661508, U16369), B. ovata (AY081192, AY603400), B. rossi (JN982353), B. vitalii (JN880430, JN880431) and B. vogeli (HM590440). In addition, the 18S rRNA sequences of 9 related protozoan species were also obtained from GenBank: Theileria equi (AB515307, AB515312), T. parva (L02366), Trypanosoma cruzi (AF303659), Toxoplasma gondii (L37415), Neosporo caninum (U63069), Eimeria arnyi (AY613853), Cytauxzoon felis (AY679105), Hepatozoon americanum (AF176836), Cryptosporidium meleagridis (AF112574) and C. parvum (L16996).Fig. 1


Development of a pan-Babesia FRET-qPCR and a survey of livestock from five Caribbean islands.

Li J, Kelly P, Zhang J, Xu C, Wang C - BMC Vet. Res. (2015)

Alignment of the partial 18S rRNA gene amplicons of Babesia spp. and other related species. The upstream primer-1 (in red), upstream primer-2 (in blue), the fluorescein/LCRed 640 probes and the downstream primer are indicated in the top of the boxes. Dots indicate nucleotides identical to the primers and probes, and the dashes denote the deletion of the nucleotides. The upstream primers and two probes are used as the indicated sequences while the downstream primer is used as antisense oligonucleotide. While the probes and downstream primer show minimum mismatch with Babesia spp. and other related species, the upstream primers (−1 and −2) have 0–1 nucleotide mismatch with Babesia spp. but 6–16 nucleotide mismatches with the related non- Babesia species
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4588467&req=5

Fig1: Alignment of the partial 18S rRNA gene amplicons of Babesia spp. and other related species. The upstream primer-1 (in red), upstream primer-2 (in blue), the fluorescein/LCRed 640 probes and the downstream primer are indicated in the top of the boxes. Dots indicate nucleotides identical to the primers and probes, and the dashes denote the deletion of the nucleotides. The upstream primers and two probes are used as the indicated sequences while the downstream primer is used as antisense oligonucleotide. While the probes and downstream primer show minimum mismatch with Babesia spp. and other related species, the upstream primers (−1 and −2) have 0–1 nucleotide mismatch with Babesia spp. but 6–16 nucleotide mismatches with the related non- Babesia species
Mentions: The 18S rRNA sequences for 22 recognized Babesia spp. of public health significance and/or veterinary importance were obtained from GenBank (Fig. 1): B. microti (AB071177, AB219802), B. leo (AF244911, AY452708), B. rodhaini (DQ641423, AB049999), B. felis (AF244912, AY452707), B. poelea (DQ200887), B. bigemina (JQ723014, KF112076), B. bovis (HQ264124, HQ264127), B. caballi (AY534883), B. canis (AY072926, JN982353), B. capreoli (FJ944828, GQ304526), B. crassa (AY260176, JX542614), B. divergens (FJ944822, FJ944826), B. gibsoni (KJ142323), B. hongkongensis (JQ867356), B. kiwiensis (EF55315), B. major (JF802040), B. motasi (AY260179, AY533147), B. odocoilei (AY661508, U16369), B. ovata (AY081192, AY603400), B. rossi (JN982353), B. vitalii (JN880430, JN880431) and B. vogeli (HM590440). In addition, the 18S rRNA sequences of 9 related protozoan species were also obtained from GenBank: Theileria equi (AB515307, AB515312), T. parva (L02366), Trypanosoma cruzi (AF303659), Toxoplasma gondii (L37415), Neosporo caninum (U63069), Eimeria arnyi (AY613853), Cytauxzoon felis (AY679105), Hepatozoon americanum (AF176836), Cryptosporidium meleagridis (AF112574) and C. parvum (L16996).Fig. 1

Bottom Line: Babesia spp. are tick-borne protozoan hemoparasites and the second most common blood-borne parasites of mammals, in particular domestic animals.We used the Clustal Multiple Alignment program and 18S rRNA gene sequences of 22 Babesia species from GenBank to develop a PCR that could detect a wide variety of Babesia spp. in a single reaction.The pan-Babesia FRET-qPCR we developed reliably detected B. gibsoni, B. canis, B. vogeli, B. microti, B. bovis, and B. divergens under controlled conditions but did not react with closely related species, mainly Hepatozoon americanum, Theileria equi, and Toxoplasma gondii.

View Article: PubMed Central - PubMed

Affiliation: Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University College of Animal Science and Technology, Yangzhou, Jiangsu, 225009, P. R. China. lijing900401@163.com.

ABSTRACT

Background: Babesia spp. are tick-borne protozoan hemoparasites and the second most common blood-borne parasites of mammals, in particular domestic animals. We used the Clustal Multiple Alignment program and 18S rRNA gene sequences of 22 Babesia species from GenBank to develop a PCR that could detect a wide variety of Babesia spp. in a single reaction. The pan-Babesia FRET-qPCR we developed reliably detected B. gibsoni, B. canis, B. vogeli, B. microti, B. bovis, and B. divergens under controlled conditions but did not react with closely related species, mainly Hepatozoon americanum, Theileria equi, and Toxoplasma gondii.

Results: When we tested the pan-Babesia FRET-qPCR on DNA of whole blood from 752 cattle, sheep, goats, donkeys and horses from five Caribbean islands, we detected Babesia spp. expected to be present in the animals, mainly B. bovis and B. bigemina in cattle and B. caballi in horses and donkeys. Further, we found that animals were not uncommonly infected with species of Babesia usually associated with other hosts, mainly B. vogeli and B. gibsoni in cattle, sheep and goats, B. rossi in goats, and B. caballi in goats and sheep. Finally, the pan-Babesia FRET-qPCR enabled us to identify unknown species of Babesia in cattle, goats, sheep and donkeys.

Conclusions: Overall, 70 % (525/752) of the animals we tested were positive confirming earlier limited studies that infections with Babesia spp. are common in livestock in the Caribbean.

No MeSH data available.


Related in: MedlinePlus