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United States Department of Agriculture

Agricultural Research Service

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Research Project: THREAT OF CATTLE FEVER IN TEXAS: SCREENING FOR THE PRESENCE OF BABESIA AND DETERMINING TICK POPULATION STRUCTURE USING MOLECULAR GENETICS

Location: Animal Diseases Research

2011 Annual Report


1a.Objectives (from AD-416)
The primary objective of this proposal is to answer basic questions regarding population structure of cattle fever ticks and prevalence of Babesia sp. in South Texas. The following questions and hypotheses will be addressed: 1) Do cattle fever ticks in the Texas quarantine zone demonstrate population structure, either locally or across the entire landscape? H1: Limited gene flow leads to high levels of population structure and is due to barriers to host dispersal (e.g. fences restrict host movement). H2: Widespread gene flow leads to low levels of population differentiation because of widespread host movement, or other mechanisms of dispersal. 2) Are cattle fever ticks on non-bovine ungulates a source of ticks on cattle? H1: Ticks use hosts indiscriminantly, and thus display no genetic structure across host species. H2: Ticks form separate populations on cattle and other wild ungulates, with little genetic admixture across host types (possibly leading to host-race formation). 3) What geographic sources gave rise to the populations(s) of cattle fever ticks in Texas, and have multiple introductions occurred? H1) Cattle fever ticks in Texas originated from a single introduction event from Mexico, have become permanently established in the quarantine zone, and display a limited subset of the total genetic variation found in Mexico. H2) Texas ticks originate from a wide variety of locations, are continually reintroduced, and retain most of the genetic variation found in Mexico. 4) What is the source of new cattle fever tick populations found outside of the permanent quarantine zone? H1) Recent tick outbreaks originate from nearby areas of the permanent quarantine zone, and therefore geographic proximity can be used to predict genetic similarity. H2) New tick outbreaks originate from random locales within the quarantine zone or from Mexico, and thus geographic proximity does not correlate with genetic similarity. 5) Do cattle fever ticks collected from cattle and non-bovine ungulates within the Texas quarantine zone carry B. bovis and B. bigemina? H1) Babesia has re-emerged in South Texas, and is found in ticks on cattle and/or wild ungulates in this region. H2) Babesia has not re-emerged in South Texas, and is not found in ungulates or cattle in this region. 6) What is the geographic origin of Babesia populations in South Texas? H1) The most likely source of Babesia in the U.S. is Mexico, resulting in high similarity between any US Babesia and pathogen strains from Mexico. H2) Babesia is transported globally, and US Babesia are not similar to strains from Mexico. 7) Do Babesia strains in the permanent quarantine zone demonstrate population structure? H1) B. bovis and B. bigemina are disseminated widely across the permanent quarantine zone, resulting in low levels of population genetic structure. H2) B. bovis and B. bigemina originate from multiple locations but have limited dispersal, resulting in strong population structuring across the quarantine zone.


1b.Approach (from AD-416)
Cattle fever ticks collected from within the permanent and temporary preventive quarantine zone and those provided as part of a collaborative effort with scientists in Mexico will be used as the basis for the proposed tick population genetic studies. Genomic DNA will be isolated from individual tick specimens and used to amplify molecular markers, both published and identified as part of the proposed project, the results of which will be analyzed using standard population biology algorithms. In addition, genomic DNA will be isolated from collected semi-engorged and engorged females for use in amplification of markers specific for Babesia bovis and B. bigemina, to assess the prevalence of the pathogen in South Texas. Existing markers will be evaluated, but the proposed project also aims to identify new species-specific markers. Isolates will be compared between counties of South Texas and also with states within Mexico to determine whether geographic differences can be identified.


3.Progress Report

Our collaborators at Northern Arizona University have genotyped >1,200 R. microplus ticks at 10 microsatellite loci. The results reveal the existence of strong genetic structure among tick populations in the Texas quarantine zone, both at the landscape scale (among counties) and the local level (within counties). Interestingly, ticks collected from cattle and white-tailed deer in the same pasture share a common gene pool. This is consistent with the possibility of deer being a source of ticks on cattle, and vice versa. Our DNA fingerprinting markers allow us to run population assignment tests to reveal the source of ticks intercepted by USDA-APHIS. We are also developing a panel of microsatellite loci for the other important cattle fever vector, R. annulatus. These genotyping approaches will provide important insights into tick dispersal and gene flow in the Texas quarantine zone. Here at the Animal Disease Research Unit in Pullman WA, we have begun to assess biological differences between populations of Rhipicephalus microplus ticks originating from Deer and from Cattle. We received larval ticks that are the offspring of engorged females collected from deer and from cattle in the Texas Quarantine zone and have carried out a vector competence study with these ticks by feeding them on Babesia bovis infected cattle. There are significant differences between deer and cattle tick populations in feeding and oviposition success, most significantly, ticks originating from cattle produce more offspring than do ticks originating from deer. The infection and transmission portions of this study are still in progress, but so far ticks from either deer or cattle appear to be equally competent vectors of B. bovis.


Last Modified: 4/17/2014
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