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United States Department of Agriculture

Agricultural Research Service

Research Project: MINING THE GENOME OF RHIPICEPHALUS MICROPLUS TO DEVELOP NOVEL CONTROL TECHNOLOGY AND VACCINES

Location: Tick and Biting Fly Research

Title: Phenotype changes inherited by crossing pyrethroid susceptible and resistant genotypes from the cattle tick Rhipicephalus (Boophilus) microplus

Authors
item Aguilar-Tipacamu, G -
item Rosario-Cruz, Rodrigo -
item Miller, Robert
item Guerrero, Felix
item Rodriguez-Vivas, R -
item Garcia-Vazquez, Z -

Submitted to: Experimental and Applied Acarology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: March 5, 2011
Publication Date: July 20, 2011
Citation: Aguilar-Tipacamu, G., Rosario-Cruz, R., Miller, R., Guerrero, F., Rodriguez-Vivas, R.I., Garcia-Vazquez, Z. 2011. Phenotype changes inherited by crossing pyrethroid susceptible and resistant genotypes from the cattle tick Rhipicephalus (Boophilus) microplus. Experimental and Applied Acarology. 54(3):301-311.

Interpretive Summary: Multiple resistance mechanisms are common in cattle ticks, Rhipicephalus microplus, from some areas of Mexico and pyrethroid resistance is especially common. In many arthropods, resistance to pyrethroids has been associated with the presence of nucleotide mutations in the sodium channel gene. A DNA-based assay is available to identify individual cattle ticks that possess a pyrethroid resistance-associated mutation in the sodium channel. The mechanisms of inheritance of the pyrethroid resistance mutation have not been studied. This study was designed to analyze changes in pyrethroid resistance obtained from reciprocal crosses between pyrethroid resistant and susceptible strains of the cattle tick from Mexico. Changes in phenotype were measured by larval packet test bioassays and changes in genotype were measured by sodium channel diagnostic PCR for the sodium channel pyrethroid resistance-associated gene mutation. The resistance to the pyrethroids was determined to be inherited as a recessive trait, although the presence of other components of metabolic resistance mechanisms or other sodium channel genes could not be discounted.

Technical Abstract: Dialelic crosses and backcrosses of pyrethroid resistant (RR) and susceptible (SS) Rhipicephalus (Boophilus) microplus tick strains were carried out and the substitution (Phe-Ile) within the sodium channel gene was monitored in order to analyze the effects of the genotype on the pyrethroid resistance phenotype as measured by the larval packet test (LPT). Parental strains: susceptible (SS) and resistant (RR); dialelic crosses: RS (Male RR X Female SS), and SR (Male SS X Female RR); and backcrosses: RS X SS, RS X RR, SR X SS and SR X RR were infested on 280 kg calves. Resistance type (monogenic or polygenic) and effective dominance were determined based on the discriminant concentration (DC) for cypermethrin (0.5%), deltamethrin (0.09%) and flumethrin (0.01%). Allele specific PCR (AS-PCR) was used for genotyping, looking at a sodium channel mutation (Phe-Ile substitution). The mortality rates and allele frequency of susceptible and pyrethroid resistant reference strains were 0% mortality and 90% RR alleles for resistant strain, and 100% mortality and 0% RR alleles as measured by the larval packet test (LPT) and allele specific PCR (AS-PCR) respectively. Backcrossed strain SR 9 RR showed an effective dominance (DML) of 0.605 for cypermethrin, 0.639 for deltamethrin and 0.498 for flumethrin, while survival of backcrosses RS X SS, RS X RR and SR X SS showed a significant tendency to recessivity. Backcrossed strain SR X RR (69.4%) also showed a higher RR genotype frequency with regards to RS X SS (25.5%), RS X RR (36.7%) and SR X SS (32.0%), however, susceptible allele was inherited in general as an incomplete dominant trait. Monogenic inheritance hypothesis was tested and the results showed monogenic inheritance for cypermethrin and flumethrin (P<0.05) but not for deltamethrin (P>0.05). However, significant correlation was found between RR genotype and the survival rate for all three pyrethroids used (P<0.05), suggesting that a single substitution on the sodium channel gene can be responsible for resistance to pyrethroids as a class, due to the high frequency for RR genotypes. Combination with different mutations or metabolic resistance mechanisms cannot be excluded.

Last Modified: 11/28/2014
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