Dataset Information

Acaricide Resistance Development in Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) Populations against Amitraz and Deltamethrin on Communal Farms of the King Sabata Dalindyebo Municipality, South Africa.

ABSTRACT: Chemical acaricides are widely used to control ticks and tick-borne pathogens in cattle. However, prolonged and indiscriminate use of these chemicals inevitably leads to the selection of resistant ticks. In-vitro bioassays (adult and larval immersion tests) were conducted to assess amitraz and deltamethrin resistance in Rhipicephalus (Boophilus) microplus populations from communal farms of the King Sabata Dalindyebo municipality of South Africa. Data generated on percentage inhibition of oviposition (%IO) revealed that all the tick populations assessed showed resistance (%IO ≤ 95%) to at least one of the acaricides. All six tick populations assessed for efficacy (%IO ≥ 95%) at the DD) with deltamethrin were resistant (%IO ≤ 95%) and only one of the six tick populations assessed for efficacy with amitraz was susceptible. Based on the resistance ratios (RR), the adult immersion test detected amitraz and deltamethrin resistance in three (RR ranging from 2.30 to 3.21) and five (RR ranging from 4.10 to 14.59) of the six tick populations, respectively. With the larval immersion test, deltamethrin and amitraz resistance (larval mortality < 90% at the DD) was detected in all four and three of four R. (B.) microplus populations assessed, respectively. These data are critical for the design of an effective and sustainable tick control strategy on the communal farms.

SUBMITTER: Dzemo WD

PROVIDER: S-EPMC10383965 | biostudies-literature | 2023 Jun

REPOSITORIES: biostudies-literature

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Acaricide Resistance Development in <i>Rhipicephalus (Boophilus) microplus</i> (Acari: Ixodidae) Populations against Amitraz and Deltamethrin on Communal Farms of the King Sabata Dalindyebo Municipality, South Africa.

Dzemo William Diymba WD Vudriko Patrick P Ramatla Tsepo T Thekisoe Oriel O

Pathogens (Basel, Switzerland) 20230626 7

Chemical acaricides are widely used to control ticks and tick-borne pathogens in cattle. However, prolonged and indiscriminate use of these chemicals inevitably leads to the selection of resistant ticks. In-vitro bioassays (adult and larval immersion tests) were conducted to assess amitraz and deltamethrin resistance in <i>Rhipicephalus (Boophilus) microplus</i> populations from communal farms of the King Sabata Dalindyebo municipality of South Africa. Data generated on percentage inhibition of ...[more]

PMID: 37513722

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Project description:Globally, the prevalence of Rhipicephalus microplus resistance to various acaricides has increased, and there is a need for the identification of molecular markers that can predict phenotypic resistance. These markers could serve as alternatives to the larval packet test (LPT), enabling rapid and accurate monitoring of resistance in these ticks against multiple acaricides. However, many of the historically identified markers are present in isolates from specific countries and their role in acaricide resistance remains unclear. This study aimed to assess these mutations by sequencing genomic regions encoding proteins historically associated with acaricide target site insensitivity and increased acaricide detoxification and comparing resistant and susceptible isolates from eight different countries. Employing a novel multiplex PCR setup developed during the study, the coding regions of 11 acaricide-resistant targets were amplified and sequenced across 37 R. microplus isolates from different locations. The identified mutations, both previously reported and novel, were compared between acaricide-susceptible and acaricide-resistant isolates, phenotypically characterized using the larval packet test or larval immersion test across five acaricide classes. Genotypes were then correlated with available phenotypes, and protein modelling of novel nonsynonymous mutations was conducted to assess their potential impact on acaricide resistance. Previously reported resistance-associated mutations were detected, some of which were present in both resistant and susceptible isolates. Novel mutations emerged from the 11 targets, but distinctions between susceptible and resistant isolates were not evident, except for the prevalent kdr mutation in synthetic pyrethroid-resistant isolates. The quest for predictive molecular markers for monitoring acaricide resistance remains challenging. Nevertheless, by utilizing a representative group of isolates, we determined that several historical mutations were present in both resistant and susceptible isolates. Additionally, the study provides valuable genetic data on acaricide-resistant and susceptible isolates from different geographical regions, focusing on genomic regions implicated in resistance. This baseline data offers a critical foundation for further research and the identification of more reliable molecular markers.

Project description:The southern cattle fever tick, Rhipicephalus (Boophilus) microplus, is the most economically important ectoparasite of cattle worldwide. A limitation for sustainable control and eradication is the emergence of acaricide resistance among tick populations. Molecular diagnostic tools offer the opportunity to detect resistance rapidly, which can be complemented with confirmatory bioassays with larvae and adult ticks that are more resource and time consuming to generate. Synthetic pyrethroid resistance is one of the most prevalent and well-studied forms of resistance in arthropods, being linked with target site alterations in the sodium ion channel gene. Here, we report research on a novel molecular method to detect mutations in the para-sodium channel gene of R. microplus associated with acaricide resistance that is based on quantitative PCR high-resolution melt (HRM) analysis. Genomic DNA fragments of domains II and III of the para-sodium channel gene were amplified by real-time PCR in the presence of EVA®Green dye to test resistant and susceptible reference ticks from the U.S., Brazil, and Mexico. Larval packet tests with discriminating doses and a modified lethal time analysis were performed to confirm resistance to permethrin, cypermethrin, deltamethrin, and flumethrin in laboratory strains. Tick specimens collected from cattle that were inspected at the United States Port-of-Entry at the Texas-Mexico border were also genotyped. Previously described mutations associated with pyrethroid resistance (T170C, C190A, G184C, and T2134A) were successfully detected by qPCR-HRM in different genotypes and confirmed by sequencing. A novel non-synonymous SNP located at domain III (C2136A) and the G215T mutation in domain II, previously described only in Asian R. microplus and R. australis, were also detected with the HRM and confirmed by sequencing. This technique could be adapted for high-throughput screening, detection, and discovery of allele-specific mutations in cattle tick outbreak populations to inform eradication strategies in the USA. This knowledge could also be applied to integrated control programs in other parts of the world where R. microplus is endemic and where similar SNPs have been identified associated with pyrethroid resistance. This study highlights the existence of several mutations in the para-sodium channel gene in different combinations in field populations of R. microplus from Mexico.

Project description:BackgroundThe southern cattle tick, Rhipicephalus (Boophilus) microplus, is an economically important parasite of cattle and can transmit several pathogenic microorganisms to its cattle host during the feeding process. Understanding the biology and genomics of R. microplus is critical to developing novel methods for controlling these ticks.ResultsWe present a global comparative genomic analysis of a gene index of R. microplus comprised of 13,643 unique transcripts assembled from 42,512 expressed sequence tags (ESTs), a significant fraction of the complement of R. microplus genes. The source material for these ESTs consisted of polyA RNA from various tissues, lifestages, and strains of R. microplus, including larvae exposed to heat, cold, host odor, and acaricide. Functional annotation using RPS-Blast analysis identified conserved protein domains in the conceptually translated gene index and assigned GO terms to those database transcripts which had informative BlastX hits. Blast Score Ratio and SimiTri analysis compared the conceptual transcriptome of the R. microplus database to other eukaryotic proteomes and EST databases, including those from 3 ticks. The most abundant protein domains in BmiGI were also analyzed by SimiTri methodology.ConclusionThese results indicate that a large fraction of BmiGI entries have no homologs in other sequenced genomes. Analysis with the PartiGene annotation pipeline showed 64% of the members of BmiGI could not be assigned GO annotation, thus minimal information is available about a significant fraction of the tick genome. This highlights the important insights in tick biology which are likely to result from a tick genome sequencing project. Global comparative analysis identified some tick genes with unexpected phylogenetic relationships which detailed analysis attributed to gene losses in some members of the animal kingdom. Some tick genes were identified which had close orthologues to mammalian genes. Members of this group would likely be poor choices as targets for development of novel tick control technology.

Dataset Information

Acaricide Resistance Development in Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) Populations against Amitraz and Deltamethrin on Communal Farms of the King Sabata Dalindyebo Municipality, South Africa.

Publications

Acaricide Resistance Development in <i>Rhipicephalus (Boophilus) microplus</i> (Acari: Ixodidae) Populations against Amitraz and Deltamethrin on Communal Farms of the King Sabata Dalindyebo Municipality, South Africa.

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