High Prevalence and Genetic Heterogeneity of <i>Anaplasma marginale</i> in Smallholder Bovine Populations of Pakistan, and Its Implications

Bovine anaplasmosis, caused by <i>Anaplasma marginale</i>, is a major tick-borne disease in tropical and subtropical regions of the world, leading to significant production losses. Prolonged convalescence periods are common and surviving animals often become subclinical carriers. This st...

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Main Authors: Abdul Ghafar, Waseem Shaukat, Muhammad Waqas, Charles G. Gauci, Robin B. Gasser, Abdul Jabbar
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Pathogens
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Online Access:https://www.mdpi.com/2076-0817/14/5/499
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author Abdul Ghafar
Waseem Shaukat
Muhammad Waqas
Charles G. Gauci
Robin B. Gasser
Abdul Jabbar
author_facet Abdul Ghafar
Waseem Shaukat
Muhammad Waqas
Charles G. Gauci
Robin B. Gasser
Abdul Jabbar
author_sort Abdul Ghafar
collection DOAJ
description Bovine anaplasmosis, caused by <i>Anaplasma marginale</i>, is a major tick-borne disease in tropical and subtropical regions of the world, leading to significant production losses. Prolonged convalescence periods are common and surviving animals often become subclinical carriers. This study aimed to detect and characterise A. marginale in bovines in smallholder dairy farms across diverse climatic zones of Pakistan using molecular methods. In total, 321 blood DNA samples from apparently healthy cattle (n = 174) and buffaloes (n = 147) from six districts in Pakistan were tested for A. marginale using a nested PCR assay, targeting part of the major surface protein B gene (msp1β) as a genetic marker, followed by agarose gel electrophoresis and selective sequencing of amplicons from test-positive samples. Of the 321 DNA samples tested, 135 (42.1%) were test-positive for A. marginale. Prevalence was significantly higher in cattle (64.4%; 112/174) than in buffaloes (15.6%; 23/147), and female bovines (43.5%; 108/248) were more frequently infected than males (37%; 27/73). Phylogenetic analysis of the msp1β sequence data (n = 42) revealed that A. marginale from Pakistan clustered with those from Brazil, Thailand, South Africa, and the USA. This study represents the first comprehensive investigation of A. marginale from bovines from diverse agroecological zones of Pakistan and will further stimulate population genetic studies of A. marginale and investigations into the economic impact of subclinical infections in bovines in smallholder farming systems.
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spelling doaj-art-dabf19d1065c4fbbb7d9a54d4356b6b52025-08-20T03:14:43ZengMDPI AGPathogens2076-08172025-05-0114549910.3390/pathogens14050499High Prevalence and Genetic Heterogeneity of <i>Anaplasma marginale</i> in Smallholder Bovine Populations of Pakistan, and Its ImplicationsAbdul Ghafar0Waseem Shaukat1Muhammad Waqas2Charles G. Gauci3Robin B. Gasser4Abdul Jabbar5Melbourne Veterinary School, The University of Melbourne, Werribee, VIC 3030, AustraliaFaculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 1N4, CanadaFaculty of Organic Agricultural Sciences, University of Kassel, 34109 Kassel, GermanyMelbourne Veterinary School, The University of Melbourne, Werribee, VIC 3030, AustraliaMelbourne Veterinary School, The University of Melbourne, Werribee, VIC 3030, AustraliaMelbourne Veterinary School, The University of Melbourne, Werribee, VIC 3030, AustraliaBovine anaplasmosis, caused by <i>Anaplasma marginale</i>, is a major tick-borne disease in tropical and subtropical regions of the world, leading to significant production losses. Prolonged convalescence periods are common and surviving animals often become subclinical carriers. This study aimed to detect and characterise A. marginale in bovines in smallholder dairy farms across diverse climatic zones of Pakistan using molecular methods. In total, 321 blood DNA samples from apparently healthy cattle (n = 174) and buffaloes (n = 147) from six districts in Pakistan were tested for A. marginale using a nested PCR assay, targeting part of the major surface protein B gene (msp1β) as a genetic marker, followed by agarose gel electrophoresis and selective sequencing of amplicons from test-positive samples. Of the 321 DNA samples tested, 135 (42.1%) were test-positive for A. marginale. Prevalence was significantly higher in cattle (64.4%; 112/174) than in buffaloes (15.6%; 23/147), and female bovines (43.5%; 108/248) were more frequently infected than males (37%; 27/73). Phylogenetic analysis of the msp1β sequence data (n = 42) revealed that A. marginale from Pakistan clustered with those from Brazil, Thailand, South Africa, and the USA. This study represents the first comprehensive investigation of A. marginale from bovines from diverse agroecological zones of Pakistan and will further stimulate population genetic studies of A. marginale and investigations into the economic impact of subclinical infections in bovines in smallholder farming systems.https://www.mdpi.com/2076-0817/14/5/499<i>Anaplasma marginale</i>anaplasmosisbovinecattleendemic stabilitymajor surface protein B gene
spellingShingle Abdul Ghafar
Waseem Shaukat
Muhammad Waqas
Charles G. Gauci
Robin B. Gasser
Abdul Jabbar
High Prevalence and Genetic Heterogeneity of <i>Anaplasma marginale</i> in Smallholder Bovine Populations of Pakistan, and Its Implications
Pathogens
<i>Anaplasma marginale</i>
anaplasmosis
bovine
cattle
endemic stability
major surface protein B gene
title High Prevalence and Genetic Heterogeneity of <i>Anaplasma marginale</i> in Smallholder Bovine Populations of Pakistan, and Its Implications
title_full High Prevalence and Genetic Heterogeneity of <i>Anaplasma marginale</i> in Smallholder Bovine Populations of Pakistan, and Its Implications
title_fullStr High Prevalence and Genetic Heterogeneity of <i>Anaplasma marginale</i> in Smallholder Bovine Populations of Pakistan, and Its Implications
title_full_unstemmed High Prevalence and Genetic Heterogeneity of <i>Anaplasma marginale</i> in Smallholder Bovine Populations of Pakistan, and Its Implications
title_short High Prevalence and Genetic Heterogeneity of <i>Anaplasma marginale</i> in Smallholder Bovine Populations of Pakistan, and Its Implications
title_sort high prevalence and genetic heterogeneity of i anaplasma marginale i in smallholder bovine populations of pakistan and its implications
topic <i>Anaplasma marginale</i>
anaplasmosis
bovine
cattle
endemic stability
major surface protein B gene
url https://www.mdpi.com/2076-0817/14/5/499
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