Five strains of Rhipicephalus (Boophilus) microplus collected from Tamaulipas Mexico were tested for resistance against several classes of acaricides commonly used. All were resistant to fipronil. Four of five were co-resistant to permethrin and coumaphos in addition to being resistant to fipronil. One strain, El Zamora was found multi-resistant to permethrin, coumaphos, fipronil, and amitraz. Selection with fipronil for 3 generations produced a resistance ratio of 8.3 and 9.4 at the LC(50) and the LC(99) estimates, respectively. Permethrin resistance in El Zamora was possibly linked to elevated esterase (CZEST9) and could be a contributing factor of resistance to fipronil. The implications of resistance for the control of the southern cattle tick in the future are discussed.

Over a 7 yr period, we monitored the effect of a commercially available, amitraz impregnated anti-tick collar in controlling free-living populations of lone star ticks, Amblyomma americanum (L.) when manually fitted around necks of white-tailed deer, Odocoileus virginianus (Zimmermann). Study animals in treatment and control groups were confined in 38.8 ha game-fenced and densely vegetated treatment plots in South Texas. Tick densities during years 1 and 7 served as untreated pre- and posttreatment comparisons and treatments occurred during years 2 through 5. Reductions in tick densities in the treatment plot were compared against tick densities in a control plot having similar vegetation and numbers of untreated deer. During years of treatment, indices of control pressure ranged from 18.2 to 82.6 for nymphs and 16.9-78.7 for adults, and efficacy, expressed as percentage control during the final year of treatment, was 77.2 and 85.0%, respectively, for nymphal and adult ticks. These data show that acaricidal collar treatments provide efficacies very similar to those achieved with the existing ivermectin-medicated bait and '4-Poster' topical treatment technologies to control ticks feeding on wild white-tailed deer.

Various acaricide-resistant strains of Rhipicephalus (Boophilus) microplus, representative of the major resistance mechanisms found in Mexico and Brazil, were exposed to spinosad using the Food and Agriculture Organization of the United Nations larval packet test and adult immersion test (AIT). Larvae of all strains tested were found to be susceptible to spinosad. Conversely, spinosad did not show toxic activity toward engorged females used in the AIT. In vitro tests against larvae, nymphs, and adults of acaricide-susceptible Amblyomma americanum and Dermacentor variabilis revealed differences in activity that were dependent on tick species and parasitic life stage. Spinosad seemed to be a viable alternative to current acaricides available for tick control in the species tested. The larval packet test should be used for future monitoring of resistance, as the AIT did not provide useful information with this chemical. The potential benefit for the use of spinosad in integrated pest management or eradication programs is discussed.

From 1907 when the fever tick eradication campaign began until 1933, the tick eradication methods of dipping cattle in an acaricide or "pasture vacation" were enormously successful in eradicating southern cattle ticks [Rhipicephalus (Boophilus) microplus (Canestrini)], until failures began to occur in some areas of Florida. Regarding the failures in Florida, the consensus was that populations of white-tailed deer [Odocoileus virginianus (Zimmermann)] infested with southern cattle ticks were responsible. After numerous deer in several counties were killed, eradication was achieved in Florida. As in Florida, in Texas increasing numbers of failures of the pasture vacation approach to tick eradication from the 1970s to the present are known to be related to the abundance of white-tailed deer and perhaps other wild ungulate species. A sizable body of evidence confirms the hypothesis that white-tailed deer support the dispersal and maintenance of both cattle ticks [Rhipicephalus (Boophilus) annulatus (Say)] and southern cattle ticks (cattle fever ticks) within the permanent quarantine or buffer zone in South Texas along the Rio Grande, as well as in the so-called free ("cattle fever tick-free") area north and east of the buffer zone and extending to the east coast of the United States. As of August 2009, in addition to the permanent quarantine zone of approximately 2233 km2, three temporary preventative or blanket quarantines were established. Currently, only two methodologies exist to control ticks feeding on white-tailed deer: (1) a systemic treatment method involving dispersal of ivermectin-medicated corn, Zea mays L.; and (2) two topical treatment methods, '4-poster' deer treatment bait stations and '2-poster' deer treatment feeder adapters, both of which passively apply topically active acaricide to deer for the eradication of populations of cattle fever tick associated with white-tailed deer. This study presents and summarizes confirmational support for the role of white-tailed deer derived from historical accounts, circumstantial evidence from review of recent infestations, and cattle fever tick infestations on white-tailed deer that were live-captured and examined specifically for cattle fever ticks.

From 1998-2002 twenty-five deer self-treatment devices (4-Posters), using 2% amitraz, were operated at three locations in Maryland to determine their effectiveness in controlling blacklegged ticks, Ixodes scapularis Say, and lone star ticks, Amblyomma americanum (L.). Each treatment site was approximately 518 ha and paired with a similar site lacking 4-Posters. Locations varied in deer density, tick abundance, and land use. Flagging for host-seeking ticks showed declines in tick populations at all treatment sites compared to control sites by the third year. By 2002, control of I. scapularis nymphs attributable to the 4-Poster intervention at the three sites was 69.0%, 75.8%, and 80%. Control of A. americanum nymphs at the two sites where they occurred was 99.5% and 95.3%. In 2003, the first posttreatment year, control of I. scapularis remained around 2001-2002 levels, but by 2004, an upward trend in nymphal numbers was detectable. Populations of A. americanum showed no increase posttreatment. These results demonstrate that control of these tick species is locally possible with 4-Poster intervention.

The efficacy of topically treating white-tailed deer with an acaricide was evaluated in a Lyme disease-endemic community of southern New York State. Twenty-four 4-Poster feeders were placed in a 5.2 km(2) treatment area in Bedford, NY, while a site in Lewisboro, NY, 4.8 km distant, served as control. Treatment periods ran from 15 September to 15 December each fall from 1997 to 2001, and from 15 March to 15 May each spring from 1998 to 2002. Corn consumption averaged 15,779 kg in fall sessions and 9054 kg in spring sessions, and a mean of 89.6% of deer in the study area showed evidence of using the feeders. Deer densities, estimated by aerial snow counts, averaged 22 and 28 deer per km(2) in Bedford and Lewisboro, respectively, over a 3-year period. Significant reductions in tick numbers on deer captured in the treatment area were noted in fall 1999 compared to deer captured at the control site. Drag sampling for nymphal host-seeking ticks indicated 63.6% control in 2001, which dropped to 54.8% the following year, but reached 80% in 2003. Higher-than-normal acorn production in 2001 that likely caused a drop in deer visitation to the feeders may have reduced efficacy against larval ticks in 2002. The 4-Poster effectively reduced the density of Ixodes scapularis, though the level of control is dependent on environmental factors that affect feeding behavior of white-tailed deer.

From 1997 to 2002, the U.S. Department of Agriculture's Northeast Area-wide Tick Control Project used acaricide-treated 4-Poster Deer Treatment Bait Stations in five eastern states to control ticks feeding on white-tailed deer. The objectives of this host-targeted technology were to reduce free-living blacklegged (Ixodes scapularis Say) and lone star (Amblyomma americanum [L.]) tick populations and thereby to reduce the risk of tick-borne disease. During 2002 to 2004, treatments were suspended, and tick population recovery rates were assayed. Subsequently, the major factors that influenced variations in efficacy were extrapolated to better understand and improve this technology. Treatments resulted in significant reductions in free-living populations of nymphal blacklegged ticks at six of the seven sites, and lone star ticks were significantly reduced at all three sites where they were present. During the study, maximal significant (p < or = 0.05) efficacies against nymphal blacklegged and lone star ticks at individual sites ranged from 60.0 to 81.7 and 90.9 to 99.5%, respectively. The major environmental factor that reduced efficacy was the occurrence of heavy acorn masts, which provided an alternative food resource for deer. Although the 4-Poster technology requires 1 or more years to show efficacy, this host-targeted intervention was demonstrated to be an efficacious, economical, safe, and environment-friendly alternative to area-wide spraying of acaricide to control free-living populations of these tick species.

The Northeast Area-wide Tick Control Project (NEATCP) was funded by the United States Department of Agriculture (USDA) as a large-scale cooperative demonstration project of the USDA-Agricultural Research Service (ARS)-patented 4-Poster tick control technology (Pound et al. 1994) involving the USDA-ARS and a consortium of universities, state agencies, and a consulting firm at research locations in the five states of Connecticut (CT), Maryland (MD), New Jersey (NJ), New York (NY), and Rhode Island (RI). The stated objective of the project was "A community-based field trial of ARS-patented tick control technology designed to reduce the risk of Lyme disease in northeastern states." Here we relate the rationale and history of the technology, a chronological listing of events leading to implementation of the project, the original protocol for selecting treatment, and control sites, and protocols for deployment of treatments, sampling, assays, data analyses, and estimates of efficacy.

As part of the Northeast Area-wide Tick Control Project (NEATCP), meta-analyses were performed using pooled data on the extent of tick-vector control achieved through seven concurrent studies, conducted within five states, using U.S. Department of Agriculture "4-Poster" devices to deliver targeted-acaricide to white-tailed deer. Although reductions in the abundance of all life-stages of Ixodes scapularis were the measured outcomes, this study focused on metrics associated with I. scapularis nymphal tick densities as this measure has consistently proven to directly correlate with human risk of acquiring Lyme disease. Since independent tick sampling schemes were undertaken at each of the five environmentally distinct study locations, a meta-analytic approach permitted estimation of a single true control-effect size for each treatment year of the NEATCP. The control-effect is expressed as the annual percent I. scapularis nymphal control most consistent with meta-analysis data for each treatment year. Our meta-analyses indicate that by the sixth treatment year, the NEATCP effectively reduced the relative density of I. scapularis nymphs by 71% on the 5.14 km(2) treatment sites, corresponding to a 71% lower relative entomologic risk index for acquiring Lyme disease.

Efficacy of a single dip treatment in coumaphos at 0.182% active ingredient was determined against all parasitic stages of Rhipicephalus (Boophilus) microplus (Canestrini) on infested cattle exposed to various levels of rainfall immediately after treatment. One group of calves remained untreated with no exposure to rainfall to serve as a negative control. A second group of cattle treated with coumaphos, but not exposed to rainfall, acted as a positive treated control. Three additional groups of coumaphos-treated cattle were exposed to 14.3, 28.6, and 42.9 mm of rainfall, respectively. In the coumaphos-treated group not exposed to a rainfall, overall mean control (99.2%) was greater and mean female engorgement weight (200 mg), egg mass weight (43 mg), and index of fecundity (IF; 2.90) were all less than any group exposed to rainfall. Although exposure to the lowest level of rainfall (14.3 mm) resulted in substantially greater control (83.7%) with lower mean egg mass weight (65 mg) and IF (62.26) than ticks exposed to 28.6 or 42.9 mm of rainfall, differences were seldom significant (P > 0.05). This suggested that higher levels of rainfall exposure adversely impacted coumaphos efficacy somewhat more than lower levels of rainfall exposure. Control remained >97% against larval ticks regardless of rainfall exposure level; however, against nymphs or adults, dramatic declines in control occurred as a result of exposure to any rainfall. Thus, the movement of coumaphos treated cattle exposed to any level of rainfall would pose a high risk of dispersing viable ticks into uninfested areas.

The 4-Poster device for the topical treatment of white-tailed deer, Odocoileus virginianus (Zimmermann), against ticks using the acaricide amitraz, was evaluated in a Lyme borreliosis endemic community in Connecticut. As part of a 5-year project from 1997 to 2002, 21-24 of the 4-Posters were distributed at residential sites in Old Lyme, CT, in a core treatment area of approximately 5.2 km(2) in fall 1997. The 4-Posters were active October to mid-December and March into May, corresponding to the peak periods of activity for adult Ixodes scapularis in this particular area. Corn consumption ranged from 361 to 4789 kg/month for October and November and 696-3130 kg/month during April. Usage of 4-Posters by deer generally was high (>90%), except during acorn masts in fall 1998 and 2001. Amitraz was applied by rollers at the estimated rate of 1.3 g active ingredient/ha/year. The abundance of host-seeking I. scapularis nymphs declined significantly (p < 0.001) in the core treatment area, as compared to a control community in Old Saybrook, CT, through 2004, over the project period from 1998 to 2003, from 9.3/100m(2) to 0.97/100m(2), rising to 1.90/100m(2) in 2004. From 1999 through 2003, there were 46.1%, 49.6%, 63.4%, 64.6%, and 70.2% reductions, respectively, in the nymphal tick population in comparison with the untreated community and initial tick abundance in 1998. Control of I. scapularis adults declined to only 19.1% in 2004; 2 years after the treatment of deer was discontinued. Differences in nymphal tick abundance between the control and core treatment area were significant in 1999 (p = 0.042) and highly significant in 2001 (p < 0.001) and 2002 (p = 0.002). The passive topical application to deer of the acaricide amitraz resulted in a significant decrease in the population of free-living I. scapularis nymphs in the treated core in Connecticut.

The Santa Luiza strain of the southern cattle tick, Boophilus microplus (Canestrini) (Acari: Ixodidae), is resistant to both permethrin and amitraz. A study was conducted at the USDA Cattle Fever Tick Research Laboratory in Texas to investigate the genetic basis of permethrin resistance with cross-mating experiments, and to determine the mechanisms of permethrin resistance through synergist bioassays and biochemical analysis of esterase profiles. The Muñoz strain, an acaricide-susceptible reference strain, was used as the susceptible parent and the Santa Luiza strain, originating in Brazil, was used as the resistant parent. The Food and Agriculture Organization larval packet test was used to measure the levels of susceptibility of larvae of the parental strains, F1, backcross, F2, and F3 generations to permethrin. Results of reciprocal crossing experiments suggested that permethrin resistance was inherited as an incomplete recessive trait. There was no significant maternal effect on larval progeny's susceptibility to permethrin in the F1 and subsequent generations. The values of the degree of dominance were estimated at -0.700 and -0.522 for the F1 larvae with resistant and susceptible female parents, respectively. Results of bioassays on larval progeny of the F1 backcrossed with the resistant parent strain and of the F2 generations suggested that one major gene was responsible for permethrin resistance in the Santa Luiza strain. Selection of F3 larvae with either permethrin or amitraz led to significantly increased resistance to both permethrin and amitraz, indicating a close linkage between genes responsible for permethrin and amitraz resistance. The possible involvement of metabolic enzymes in permethrin resistance in the Santa Luiza strain of B. microplus was dismissed by the lack of enhanced synergism by TPP or PBO, as observed in synergist bioassays, as well as by the lack of enhanced esterase activity in the Santa Luiza strain relative to the susceptible strain. The results of this study suggest that other mechanisms, including a possible new sodium channel mutation that is different from the one currently known, may be responsible for permethrin resistance in the Santa Luiza strain of B. microplus.

Boophilus microplus, collected from Nuevo Leon, Mexico, were found to be highly resistant to diazinon but not highly resistant to coumaphos, suggesting that different mechanisms of resistance were present in these ticks than other Mexican organophosphate (OP)-resistant ticks reported previously. When exposed to coumaphos and piperonyl butoxide or triphenylphosphate, the LCso estimate was reduced by 3.5- and 6.3-fold, respectively, suggesting that mono-oxygenases and/or esterases were involved in resistance to coumaphos. Additionally, it was determined that this strain had an Acetylycholinesterase (AChe) that was insensitive to the active form of coumaphos, coroxon, taking at least 24 min longer to reach 50% reduction in AChE activity compared with the susceptible strain. When exposed to diazinon, none of the synergists tested significantly lowered the LC50. However, it was determined that it took six times longer to reach 60% inhibition of AChE in the resistant strain compared with the susceptible strain when exposed to the active form of diazinon, diazoxon. Insensitive AChE seems to be very common in OP-resistant B. microplus. The potential benefits for the development of a field-portable AChE inhibition assay kit are discussed.

Acaricide resistance is a major problem that hinders the control of the tropical cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini), in many parts of the world where cattle production continues to suffer severe economic losses to tick infestation. Deltamethrin and amitraz have been used alone to control R. microplus in New Caledonia for the past decade, and tick populations have developed resistance to both acaricides. A study was conducted to evaluate the effectiveness of deltamethrin and amitraz mixtures, through in vitro laboratory bioassays and in vivo on-animal efficacy trials, for the control of resistant R. microplus on cattle at two dairy farms in New Caledonia. Results of laboratory bioassays using modified larval packet tests (LPT) revealed up to 16.59-fold resistance to deltamethrin, and up to 5.86-fold resistance to amitraz. Significant synergism was observed when amitraz was used as a synergist in deltamethrin bioassays. Amitraz significantly increased deltamethrin toxicity to tick larvae, while deltamethrin was much less effective on amitraz toxicity. Synergism of amitraz by deltamethrin only occurred when the deltamethrin concentration was relatively high. Results of on animal efficacy trials of deltamethrin and amitraz alone and mixtures of both at different concentrations revealed a similar pattern of synergism. Adding amitraz to a deltamethrin formulation led to dramatic increases of percent reduction of both immature and adult ticks. In contrast, adding deltamethrin to an amitraz formulation did not increase control efficacy. Results from this study may lead to the adoption of an acaricide mixture strategy for the control of pyrethroid-resistant R. microplus in New Caledonia and elsewhere.

Selection pressure with the acaricide, amitraz was applied to a strain of Rhipicephalus (Boophilus) microplus (Canestrini) in 13 out of 18 generations, resulting in a 28.7-fold increase in resistance, indicating a shift in the phenotypic composition of the ticks, from a heterogenous mixture of both susceptible and resistant ticks when selection began to a homogenous composition of predominantly resistant ticks when selection ended. Resistant larvae of the last selected generation were infested on cattle three separate times at 20, 13, and 6 d (7 d intervals) before being dipped in a total immersion vat charged at 0.02, 0.044, and 0.096% active ingredient (AI) amitraz. There was a dose related, though not always significant, response to amitraz whereby each increase in concentration produced increased adverse effects. The number of ticks and index of fecundity (IF) of females recovered from cattle treated at 0.02 and 0.044% AI were not different from that of untreated ticks. However, female weight and egg mass weight of females recovered from cattle treated at 0.044% were lower than untreated females or females treated at 0.02% AI. All measured parameters obtained from ticks recovered from cattle treated at 0.096% AI were significantly lower than the untreated control group, but tick numbers and IF were not different from the other treated groups. At 0.02% AI the level of control was significantly lower (34.6%) than treatment at 0.096% AI (81.2%), while the level of control at 0.044% AI (57.6%) was intermediate between the other treated groups. Therefore, none of the amitraz treatments provided the required 99% control necessary for use in the U.S. Cattle Fever Tick Eradication Program, and the estimated line derived from regression analysis showed that a concentration of ca. 0.2% AI amitraz would be needed to achieve 99% control of the resistant ticks.

During the nineteenth century, as the number of cattle in the world was increased to feed the human populations of recently industrialized nations, there was a growing awareness of the relationship between infestations of cattle with ticks and disastrous epizootics of disease in herds of cattle. Problems with tick-borne diseases were related to the introduction of improved breeds of cattle into tick-infested areas because of their greater productivity than well-adapted indigenous breeds. Also, cattle infested with ticks and infected with tick-borne disease agents were moved into areas where these tick species had not previously existed (Shaw, 1969).

There are many exotic animal pathogens throughout the world that, if introduced into the United States, could have a significant detrimental impact on the health of livestock, agricultural economy, the environment, and public health. Many of these pathogens are arthropod-borne and potential vectors are readily available in the United States. A number of these arthropod-borne pathogens are discussed here as examples that illustrate the potential risk and the consequences of inadvertent introductions. Several International agencies have a role in global surveillance and in controlling animal diseases should they begin to expand their range. The risk to the United States is considerable. We propose that the United States invest in the improved infrastructure needed to reduce the risk of foreign arthropod-borne pathogens. Current U.S. programs focus on the exclusion of pathogens through regulation of animal movements and products, surveillance, especially trained animal disease diagnosticians, research support, international cooperation and, should pathogens enter our country, the resources for their prompt eradication. We suggest that the United States needs to develop a comprehensive, updated strategic plan to assess all aspects of current and future requirements, objectives, and resources needed to protect its national interests.

The sudden death of several cattle infested experimentally with Rhipicephalus (Boophilus) microplus led to a clinical investigation into the reasons for the unexpected mortality. Microscopic evidence for Babesia bigemina infection was found in blood smears from the affected animals and a PCR assay was designed to detect the presence of B. bigemina and Babesia bovis in all R. microplus strains received and propagated at the laboratory. The assay utilizes a nested PCR approach with the first PCR amplifying a well-conserved segment from the Babesia 18S ribosomal RNA gene followed by a nested PCR with Babesia speciesspecific primers and annealing temperatures enabling amplification of the 18S ribosomal RNA gene fragment specific to either B. bigemina or B. bovis. DNA from groups of 50 larvae was extracted using a rapid DNA preparation protocol, which consisted of grindingthe frozen tick larvae in PCR buffer and boiling the mixture for 5 min. The assay sensitivity allowed for the detection of the equivalent of a single infected tick larva. R. microplus eggs were also analyzed, but yolk protein viscosity created inconsistent results with the crush and boil DNA isolation protocol, necessitating the use of a more extensive proteinase K digestion-based DNA purificationmethod. We detected the presence ofB. bigeminain all strains ofR. micropluscurrently reared at the laboratory and 4of 26 strains collected from infestation outbreaks in Texas by the U.S. Cattle Fever Tick Eradication Program. Published by Elsevier B.V.

Southern cattle ticks, Boophilus microplus (Canestrini) (Acari: Ixodidae), collected in Coahuila, Mexico, were determined to be resistant to permethrin. Discriminating concentration (DC) tests at the LC99 and 2X the LC99 of susceptible ticks produced 0 and 0.5% mortality, respectively, for permethrin. However, measured mortalities for coumaphos and amitraz acaricides were within the expected ranges. Polymerase chain reaction analysis of this strain detected a 99% frequency of homozygous resistant individuals in this strain.