Population Genetic Analysis of the Theileria annulata Parasites Identified Limited Diversity and Multiplicity of Infection in the Vaccine From India

Roy, Sonti; Bhandari, Vasundhra; Barman, Madhumanti; Kumar, Pankaj; Bhanot, Vandna; Arora, Jaspreet Singh; Singh, Sanjeev; Sharma, Paresh

doi:10.3389/fmicb.2020.579929

Cited by 17 publications

(18 citation statements)

References 44 publications

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“…Genetic diversity is considered a raw material for the evolution of organisms [29]. The long-term survival of T. annulata in host animals is facilitated by the protozoan's genetic diversity, which helps the parasite escape the host's immune response [31]. T. annulata achieves this genetic diversity through chromosomal recombination in tick vectors during its sexual reproduction.…”

Section: Discussionmentioning

confidence: 99%

Molecular Epidemiology of Theileria annulata in Cattle from Two Districts in Punjab (Pakistan)

Parveen

Alkhaibari

Asif

et al. 2021

Animals

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The present study was designed to report the molecular prevalence of T. annulata in cattle blood samples collected from Punjab in Pakistan. A total of 428 cattle blood samples were collected from Districts Lodhran (n = 218) and Dera Ghazi Khan (n = 210). The prevalence of T. annulata was determined by the amplification of a fragment from its cytochrome b gene and parasite prevalence was significantly higher (p = 0.03) in the blood samples of cattle collected from Dera Ghazi Khan (70/210; 33%) as compared to Lodhran (52/218; 24%). Presence of T. annulata was also confirmed by the amplification of a fragment from their 30 kDa gene. The amplified PCR products of both genes were confirmed by DNA sequencing and these partial DNA sequences were submitted to GenBank. Phylogenetic analysis revealed that amplified partial gene sequences resembled previously reported T. annulata sequences in cattle from India, China, Iran, Tunisia, Turkey and Egypt. The incidence of T. annulata infection was higher in Sahiwal cattle (p = 0.04) than the other enrolled cattle breed from Dera Ghazi Khan. Female cattle from Lodhran (p = 0.02), while males (p = 0.02), animals housed in close compounds (p = 0.04), animals with a tick burden (p = 0.005) and farms with only cattle (p = 0.01) in Dear Ghazi Khan were found to be more susceptible to T. annulata infection. We recommend that large-scale tick and tick-borne disease control strategies be implemented in both districts under investigation, especially in Dera Ghazi Khan.

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Section: Discussionmentioning

confidence: 99%

Molecular Epidemiology of Theileria annulata in Cattle from Two Districts in Punjab (Pakistan)

Parveen

Alkhaibari

Asif

et al. 2021

Animals

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“…Tick transmitted diseases with high infection in the bovine population of Bihar with Theileria spp., A marginale and B. bigemina and their co-infection has been frequently reported (Kala et al 2018;Kumar et al 2021;Prabhakaran et al 2021;Roy et al 2021). The study indicated that the endemic region had high (76.67%) infection of tick-transmitted hemo-parasite in selected crossbred cattle based on conventional Giemsa blood smear examination (Fig 1).…”

Section: Resultsmentioning

confidence: 75%

Development of Novel Multiplex PCR for Diagnosis of Co-infected Hemo-parasites in Cattle

Kumar

Sarma

et al. 2021

IJAR

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Background: A novel, rapid and specific multiplex polymerase chain reaction was developed to diagnose hemo-parasitic infection in bovine blood co-infected with three of the most common hemo-parasites. Methods: The diagnostic process relied on the detection of the three different bovine hemoparasites isolated from red blood cells (RBCs) of cattle (N=30) by conventional Giemsa stained blood smear (GSBS) and confirmed by multiplex PCR. The multiplex PCR system was used to diagnose GSBS positive blood samples (N=12) found infected or co-infected with hemoparasites. The designed multiplex primer sets was attempted to amplify 205, 313 and 422 bp fragments of apocytochrome b, sporozoite and macroschizont 2 (spm2) and 16S rRNA gene for Babesia bigemina, Theileria annulata and Anaplasma marginale, respectively. Result: This multiplex PCR was sensitive with the ability to detect the presence of 150 ng of genomic DNA. The primers used in this multiplex PCR also showed highly specific amplification of specific gene fragments of each respective parasite. Comparing the two detection methods revealed that 58.33% of specimens showed concordant diagnoses with both techniques. The specificity, positive predictive value and kappa coefficient of the agreement was highest for diagnosis of B. bigemina and lowest for A. marginale. The overall Kappa coefficient for diagnosis based on GSBS for multiple pathogens compared to multiplex PCR was 0.56, slightly behind the threshold of 0.6 of agreement. Therefore, confirmation should always be based on PCR to rule out false positives due to differences in subjective observations, stain particles and false negatives due to low parasitemia. The simplicity and rapidity of this specific multiplex PCR method make it suitable for large-scale epidemiological studies and follow-up of drug treatments.

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“…Tick transmitted diseases with high infection in bovine population of Bihar with Theileria spp., A marginale , and B. bigemina and their co-infection has been frequently reported (Kala et al 2018; Kumar et al 2019; Kumar et al 2021; Prabhakaran et al 2021; Roy 2021). Our finding also suggest high rate of infection of these hemo-parasite and association of more than one type parasite in the animals screened by GSBS.…”

Section: Discussionmentioning

confidence: 99%

Development Of Novel Multiplex PCR For Rapid Diagnosis Of Coinfected Hemo-Parasites In Cattle

Kumar¹,

Kumar²,

Sarma³

et al. 2021

Preprint

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A novel, rapid and specific multiplex polymerase chain reaction has been developed for the diagnosis of hemo-parasitic infection in bovine blood by three of the most common hemo-parasites. The reported method relied on the detection of the three different bovine hemoparasites isolated from red blood cells (RBCs) of cattle by conventional Giemsa stained blood smear (GSBS) and confirmed by multiplex PCR. The designed multiplex primer sets can amplify 205, 313 and 422 bp fragments of apocytochrome b, sporozoite and macroschizont 2 (spm2) and 16S rRNA gene for Babesia bigemina, Theileria annulata and Anaplasma marginale, respectively. This multiplex PCR was sensitive with the ability to detect the presence of 150 ng of genomic DNA. The primers used in this multiplex PCR also showed highly specific amplification of specific gene fragments of each respective parasite DNA without the presence of non-specific and non-target PCR products. This multiplex PCR system was used to diagnose GSBS confirmed blood samples (N=12) found infected or co-infected with hemoparasites. A comparison of the two detection methods revealed that 58.33% of specimens showed concordant diagnoses with both techniques. The specificity, positive predictive value and kappa coefficient of agreement was highest for diagnosis of B. bigemina and lowest for A. marginale. The overall Kappa coefficient for diagnosis based on GSBS for multiple pathogen compared to multiplex PCR was 0.56 slightly behind the threshold of 0.6 of agreement. Therefore, confirmation should always be made based on PCR to rule out false positive due to differences in subjective observations, stain particles and false negative due to low level of parasitaemia. The simplicity and rapidity of this specific multiplex PCR method make it suitable for large-scale epidemiological studies and for follow-up of drug treatments.

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Population Genetic Analysis of the Theileria annulata Parasites Identified Limited Diversity and Multiplicity of Infection in the Vaccine From India

Cited by 17 publications

References 44 publications

Molecular Epidemiology of Theileria annulata in Cattle from Two Districts in Punjab (Pakistan)

Molecular Epidemiology of Theileria annulata in Cattle from Two Districts in Punjab (Pakistan)

Development of Novel Multiplex PCR for Diagnosis of Co-infected Hemo-parasites in Cattle

Development Of Novel Multiplex PCR For Rapid Diagnosis Of Coinfected Hemo-Parasites In Cattle

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