Perspectives on the potential of entomopathogenic fungi in biological control of ticks

Ticks are serious health threats for humans, and both domestic and wild animals. Ticks are controlled mostly by application of chemical products; but these acaricides have several negative side effects, including toxicity to animals, environmental contamination, and induction of chemical resistance in some tick populations. Entomopathogenic fungi infect arthropods in nature and can occur at enzootic or epizootic levels in their host populations. Laboratory studies clearly demonstrate that these fungi can cause high mortality in all developmental stages of several tick species, and also reduce oviposition of infected engorged females. Tick mortality following application of fungi in the field, however, often is less than that suggested by laboratory tests. This is due to many negative biotic and climatic factors. To increase efficacy of fungal agents for biological control of ticks under natural conditions, several points need consideration: (1) select effective isolates (viz., high virulence; and tolerance to high temperature, ultraviolet radiation and desiccation); (2) understand the main factors that affect virulence of fungal isolates to their target arthropods including the role of toxic metabolites of the fungal isolates; and (3) define with more precision the immune response of ticks to infection by entomopathogenic fungi. The current study reviews recent literature on biological control of ticks, and comments on the relevance of these results to advancing the development of fungal biocontrol agents, including improving formulation of fungal spores for use in tick control, and using entomopathogenic fungi in integrated pest (tick) management programs.     

West African Cattle Farmers’ Perception of Tick-Borne Diseases

Worldwide, cattle production is struggling to face the negative impacts caused by ticks and Rhipicephalus (Boophilus) microplus is one of the most harmful ticks for livestock. Most of the people in West Africa depend on cattle farming and subsistence agriculture. The presence of ticks on cattle is a major problem faced by smallholder farmers who fight for their livelihood. National and regional tick control programs could assist these rural communities in protecting their livelihoods against ticks and tick-borne diseases, but only if they take into account the targeted herders and their perception on cattle management and tick control. This paper aims to provide a better insight in the socio-economic characteristics of Beninese cattle farmers, and their perception on tick burden, as well as to document common tick control strategies. Different tick species and their seasonality are well understood by cattle herders. For tick control, many still use manual tick removal, especially in the north of the country. The high cost of acaricides, the lack of financial means of African farmers, and of the local stockbreeders in particular, limits the use of acaricides in livestock breeding in Benin. While aiming to increase the meat or milk production of their animals, stockbreeders who can afford it sometimes turn to an abusive use of acaricides, which might in time lead to an increase in tick resistance. This study remains one of the rare studies to report extensively on the perceptions of West African cattle herders.     

Methods currently used for the control of multi-host ticks: their validity and proposals for future control strategies

Ticks have a world distribution and pest status reputation of hampering livestock production through transmission of fatal disease such as theileriosis, anaplasmosis, babesiosis, etc. and bites which cause blood loss, pain and other debilitating effects. Control of ticks is largely more effective on the host. The multi-host ticks spend most of the time off the host with short feeding periods of between four and ten days. Chemical control using dips or sprays has been the traditional method of attempting to kill these ticks during the infestation period. In many situations control, using acaricides, has been quite successful. This has been possible through correct timing of the ticks' seasonal activity and feeding periods to determine the application interval, and efficacy of the acaricides. However, the rising costs of acaricides have made it almost impossible to use these chemicals on a regular basis according to the pest problem. This is particularly true in many Third World tropical countries in which tick-associated problems are more pronounced. This has necessitated the search for alternative tick control methods on an integrated approach to pest management. For this reason, vaccination against ticks and breeding for host resistance against ticks are being studied in the hope that future control strategies will involve only the economically effective acaricide application in conjunction with these and other methods.     

Strategic tick control in Burundi

Strategic tick control is an attempt to control ticks and reduce losses in animal production due to tick infestations while decreasing the cost for this control. Tick control is understood as a necessity to achieve maximum animal production from animals raised in tick-infested areas. At the same time, it has become difficult to justify intensive, year-round tick control for local breeds and their crosses. The increasing costs of acaricides, maintenance of the infrastructures, salaries for personnel, etc. have created a tick control programme where the benefits may not be adequate to justify the expense. This has been the case in Burundi. This programme with adequate participation from the cattle owners of Burundi and organizational support from government personnel can help solve this economic problem. The strategic programme reduces the period of tick control to four months each year. Once optimal participation is achieved, this period can probably be reduced to three months (Kaiser et al., 1988). The period of treatment corresponds to when there is maximal feeding activity of adult female ticks on the bovine population within a certain region. Cattle should be treated once each week throughout the four-month period in order to prevent female tick engorgement (based on the life cycle of R. appendiculatus). This will greatly reduce the number of female ticks which are available to recycle the population of ticks in the participating area. At the same time, a certain population of ticks must be maintained in order to assure contact between the cattle, ticks, and tick-borne diseases of this particular region.(ABSTRACT TRUNCATED AT 250 WORDS)     

Sheep tick Ixodes ricinus management on Welsh hill farms of designated conservation importance: implications for nationally declining birds

Impacts of sheep ticks Ixodes ricinus on livestock, gamebirds and wildlife are of concern across Europe. The present study describes livestock and tick management by 36 farmers from three upland sites of conservation importance in North Wales, where farmers consider that ticks have increased during the last 25 years. Sheep, average densities of 2.0 animals per ha were treated with pour‐on acaricides in spring, again in July, and also when removed from the moor in autumn. Given acaricide efficacy rates, sheep were susceptible to tick bites for half the period on the moor. Sheep from 17 farms were examined for ticks. Infestations were similar between farms and in relation to the acaricide used, averaging 9.3 ticks per sheep, although they were lower where the interval between successive acaricide treatments was shorter. Repeated sampling of sheep and red grouse chicks showed no annual difference in tick burdens on grouse chicks, which averaged 6.2 ticks per chick, although there were three‐fold fewer ticks on sheep in 2018 than in previous years. Tick bite rates on sheep and grouse were higher than elsewhere in the U.K. Most farmers interviewed would aim to improve their tick management using longer‐lasting acaricides and treating sheep more frequently, although they would need advice and financial help, which is currently unavailable via Government funded agri‐environment schemes.     

Auto-dissemination of commercially available fungal pathogens in a laboratory assay for management of the brown dog tick Rhipicephalus sanguineus

Rhipicephalus sanguineus is a canine tick that infests dogs throughout the world and is frequently found in homes and dog kennels. Management of this tick species is complicated by the presence of resistance to commonly utilized acaricides. Fungal formulations could provide a valuable alternative tool for management and are especially relevant indoors where detrimental environmental effects on fungal spores are of less concern. Two commercially available fungal formulations, one containing Metarhizium anisopliae and the other containing Beauveria bassiana, were compared for time to death and sporulation in nymphal ticks exposed for 60 min in treated filter paper packets. Beauveria bassiana exposure killed ticks faster than M. anisopliae exposure and B. bassiana was more likely to sporulate on tick cadavers than M. anisopliae. To determine whether infected ticks could disseminate fungus to their conspecifics, ticks were marked and treated with fungus before being placed with untreated ticks. Fungus was successfully transmitted from treated to untreated ticks. Mortality of ticks exposed to B. bassiana-exposed conspecifics occurred sooner than for those exposed to M. anisopliae-exposed conspecifics, indicating faster dissemination in the former. Therefore, although both formulations resulted in decreased longevity of ticks compared with the controls, the B. bassiana formulation holds the most promise for direct or indirect application with respect to brown dog tick management.     

Amblyomma variegatum and associated diseases in the Caribbean: strategies for control and eradication in Guadeloupe

Amblyomma variegatum, introduced into Guadelopue 150 years ago, has recently spread in the Lesser Antilles and threatens North America. Two important diseases are associated with the tick: heartwater and dermatophilosis. Heartwater occurs in Guadeloupe, Marie-Galante and Antigua; acute dermatophilosis exists almost everywhere the tick is present. Both are very pathogenic for upgraded or newly-infested cattle. After 15 years of A. variegatum control in Guadeloupe, a balance is established between a tick population maintained at a low level by spraying acaricides and an animal population either completely resistant to heartwater and dermatophilosis (creole cattle) or regularly treated against ticks (upgraded cattle). An eradication campaign against A. variegatum in Guadeloupe, to avoid the spread of the associated diseases, appears technically difficult but possible, economically profitable, but socially completely utopian. We suggest for Guadeloupe a reinforced control of the tick with a persistent acaricide, hoping that a general decrease of the tick infestation rate on cattle will eliminate heartwater and acute dermatophilosis.     

Association of BoLA-DRB3.2 alleles with tick (Boophilus microplus) resistance in cattle

Losses caused by bovine tick burdens in tropical countries have a tremendous economic impact on production systems. Besides reducing production, this parasite can cause death in the most susceptible animals. The use of commercial acaricides has been the major method of control, but their misuse has led to tick resistance to many chemicals. More recently, vaccines have been used in some countries without solving the problem completely. An alternative could be the development of resistant animals and the use of genetic markers and candidate genes that could help with the enormous task of selecting resistant animals. The bovine lymphocyte antigen genes (BoLA) have been shown to be associated with some parasitic infestations and disease incidence. Thus, the objective of the present study was to determine the association of BoLA-DRB3.2 alleles with tick resistance in cattle. The study was conducted on 231 F2 (Gyr x Holstein) animals that were artificially infested with 10,000 tick larvae. Log of tick count +1 was used as the dependent variable in a mixed animal model with allele substitution effects in addition to fixed effects of year and season at tick count, sex of calves, age of animal at tick count, hair type (short-straight, short-curl, long-straight, and long-curl), coat color (white, >75% white, 50- 75% white, and 25-50% white), and additive genetic, permanent environmental and residual effects as random. Females showed fewer ticks than males. Animals with short-straight hair were more resistant to tick infestation than animals with long-curl hair, and animals with whiter coat color also had fewer ticks. An association between BoLA alleles and lower tick number was found for alleles DRB3.2 *18, *20 and *27 at the 5% significance level. Also, one allele (DRB3.2*16) showed an association at the 10% level. Allele *27 was the most frequent in the population (30.7%), followed by alleles *16 (10.8%), *20 (8.7%) and *18 (2.4%). These results suggest that BoLA-DRB3.2 alleles could be used to help in the selection of animals resistant to tick infestation. However, further studies involving a larger population of cattle in combination with other BoLA genes may help to understand the mechanisms of resistance to parasites.     

Towards a permanent solution for controlling cattle ticks

Acaricides are essential in the short-term but do not offer a permanent solution to tick control. This situation will not change without a change of approach. A vaccine against Boophilus microplus confers partial long-term control but has little immediate effect on tick burdens. The effectiveness of acaricides and vaccination is greatest for breeds of high tick resistance. High host resistance is the key to effective long-term tick control with total resistance the ultimate aim. While improvements to acaricides and vaccines are continuously pursued, improvements to the most important single factor controlling ticks, host resistance, have been neglected. Resistance is as heritable as milk yield or growth and in tropical breeds can be increased to very high levels by selection. Despite this there are no current examples of sustained selection for tick resistance. Temperate breeds have low resistance but because of high production potentials are favoured for crossbreeding with tropical breeds. This perpetuates the need for reliance on acaricides. Selection to increase polygenic resistance of temperate breeds is impractical. However, a quantum increase can be achieved by introgressing major resistance genes. Such a gene occurs in the Belmont Adaptaur and in suitable genetic backgrounds confers 100% resistance. Total resistance is achievable and provides a permanent solution to ticks.     

The leucokinin-like peptide receptor from the cattle fever tick,Rhipicephalus microplus,is localized in the midgut periphery and receptor silencing with validated double-stranded RNAs causes a reproductive fitness cost

The cattle fever tick, Rhipicephalus microplus (Canestrini) (Acari: Ixodidae), is a one-host tick that infests primarily cattle in tropical and sub-tropical regions of the world. This species transmits deadly cattle pathogens, especially Babesia spp., for which a recombinant vaccine is not available. Therefore, disease control depends on tick vector control. Although R. microplus was eradicated in the USA, tick populations in Mexico and South America have acquired resistance to many of the applied acaricides. Recent acaricide-resistant tick reintroductions detected in the U.S. underscore the need for novel tick control methods. The octopamine and tyramine/octopamine receptors, both G protein-coupled receptors (GPCR), are believed to be the main molecular targets of the acaricide amitraz. This provides the proof of principle that investigating tick GPCRs, especially those that are invertebrate-specific, may be a feasible strategy for discovering novel targets and subsequently new anti-tick compounds. The R. microplus leucokinin-like peptide receptor (LKR), also known as the myokinin- or kinin receptor, is such a GPCR. While the receptor was previously characterized in vitro, the function of the leucokinin signaling system in ticks remains unknown. In this work, the LKR was immunolocalized to the periphery of the female midgut and silenced through RNA interference (RNAi) in females. To optimize RNAi experiments, a dual-luciferase system was developed to determine the silencing efficiency of LKR-double stranded RNA (dsRNA) constructs prior to testing those in ticks placed on cattle. This assay identified two effective dsRNAs. Silencing of the LKR with these two validated dsRNA constructs was verified by quantitative real time PCR (qRT-PCR) of female tick dissected tissues. Silencing was significant in midguts and carcasses. Silencing caused decreases in weights of egg masses and in the percentages of eggs hatched per egg mass, as well as delays in time to oviposition and egg hatching. A role of the kinin receptor in tick reproduction is apparent.     

Determination of LC50 of permethrin acaricide in semi-engorged females of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae)

Currently, the most efficient and widely used method of tick control still is the treatment with acaricides, especially permethrin (active ingredient of the Advantage^® Max3, Bayer), a pyrethroid with neurotoxic action. Due to the wide use of this acaricide in the control of the tick Rhipicephalus sanguineus, this study carried out laboratorial procedures to determine the LC₅₀ (lethal concentration fifty) of permethrin in semi-engorged females of R. sanguineus. Based on the result of 14 dilutions of permethrin in distilled water and later Probit analysis, the LC₅₀ of permethrin for R. sanguineus was 2062 ppm (1549-2675 ppm). This work can be used as a protocol with other chemicals, to determinate the LC₅₀, basic procedure for studies of control, resistance and behavior of ticks treated with acaricides, especially the brown dog tick R. sanguineus. Also, the knowledge of the LC₅₀ provides information on the potency of chemicals, the sensitivity of Arthropods to them and even estimates on pest control.     

Feeding of Ticks on Animals for Transmission and Xenodiagnosis in Lyme Disease Research

Transmission of the etiologic agent of Lyme disease, Borrelia burgdorferi, occurs by the attachment and blood feeding of Ixodes species ticks on mammalian hosts. In nature, this zoonotic bacterial pathogen may use a variety of reservoir hosts, but the white-footed mouse (Peromyscus leucopus) is the primary reservoir for larval and nymphal ticks in North America. Humans are incidental hosts most frequently infected with B. burgdorferi by the bite of ticks in the nymphal stage. B. burgdorferi adapts to its hosts throughout the enzootic cycle, so the ability to explore the functions of these spirochetes and their effects on mammalian hosts requires the use of tick feeding. In addition, the technique of xenodiagnosis (using the natural vector for detection and recovery of an infectious agent) has been useful in studies of cryptic infection. In order to obtain nymphal ticks that harbor B. burgdorferi, ticks are fed live spirochetes in culture through capillary tubes. Two animal models, mice and nonhuman primates, are most commonly used for Lyme disease studies involving tick feeding. We demonstrate the methods by which these ticks can be fed upon, and recovered from animals for either infection or xenodiagnosis.     

Detecting Resistance to Organophosphates and Carbamates in the Cattle Tick Boophilus Microplus, with a Propoxur-based Biochemical Test

Rapid and sensitive detection of resistance to insecticides in arthropods is needed. In the cattle tick, Boophilus microplus, resistance to a variety of acaricides is widespread. The most commonly used assay for resistance, the larval packet test, takes at least two, but generally six weeks for a one-host tick like B. microplus to complete and may take up to three months to complete for three-host ticks. Here we describe a test for resistance to organophosphate acaricides that can be used on larvae and adult ticks which takes less than 24 hours. The test measures the difference in acetylcholinesterase (AChE) activity in homogenates of ticks in the presence and absence of propoxur, a carbamate acaricide. We found clear discrimination of organophosphate-susceptible and organophosphate-resistant adults with 100 M propoxur. AChE from susceptible ticks had almost no activity at this concentration of propoxur whereas AChE from resistant ticks had 67% of its potential activity. AChE from heterozygote ticks could also be distinguished from AChE from homozygous-susceptible and homozygous-resistant ticks. This is the first biochemical test for resistance to an acaricide. Rapid, sensitive tests like ours will allow resistance to organophosphates to be detected soon after it develops in the field, thus, the spread of resistance might be slowed and the useful life of acaricides extended.     

The efficiency of patch sampling for determination of relative tick burdens in comparison with total tick counts

Quanttative data on host tick burdens ar fundamental for the initiation of control strategies and effective management of wildlife populations, but the methods of live sampling employed for domestic animals are unsuitable for sampling wild animals. Despite advances in the use of destructive methods (the scrub and digestion techniques) to obtain measures of the total tick burden on wildlife, these methods are too involved for many field workers, who often need only measures of relative tick burden. Recently, patch sampling methods have been introduced whereby only certain predilection sites are sampled, the presumption being that the number of ticks collected gives an indiccation of the relative degree of infestation. We examined the validity of patch sampling as a measure of relative tick burden by comparing adult ticks collected from the ears, head, neck, foreleg and perianal region of impala (aepyceros melampus) with total tick burdens of the same animals derived from the digestion technique. Adult ticks from patch sampling were positively and significantly correlated with total adults and total ticks (larvae, nymphs, and adults) on impala, with ticks patch sampled from the neck showing the highest correlation with the total tick burden. Comparison of relative tick loads from patch sampling with absolute tick loads from digestion for three classes of impala (females, bachelor males and territorial males) gave qualitatively similar results. We conclude that, when measures of relative tick load are sufficient and destructive sampling is not feasible, patch sampling can provide reliable information on relative tick burdens that are positively correlated with the total tick burden.     

Managing mosquitoes and ticks in a rapidly changing world – Facts and trends

Vector-borne diseases transmitted by mosquitoes and ticks are on the rise. The effective and sustainable control of these arthropod vectors is a puzzling challenge for public health worldwide. In the present review, I attempted to provide a concise and updated overview of the current mosquito and tick research scenario. The wide array of control tools recently developed has been considered, with special reference to those approved by the World Health Organization Vector Control Advisory Group (WHO VCAG), as well as novel ones with an extremely promising potential to be exploited in vector control programs. Concerning mosquitoes, a major focus has been given on genetically modified vectors, eave tubes, attractive toxic sugar baits (ATSB) and biocontrol agents. Regarding ticks, the recent development of highly effective repellents and acaricides (including nanoformulated ones) as well as behavior-based control tools, has been highlighted. In the second part of the review, key research questions about biology and control of mosquitoes and ticks have been critically formulated. A timely research agenda outlining hot issues to be addressed in mosquito and tick research is provided. Overall, it is expected that the present review will contribute to boost research and applications on successful mosquito and tick control strategies, along with an improved knowledge of their biology and ecology.     

Evaluating the economic damage threshold for bont tick (Amblyomma hebraeum) control in Zimbabwe

Controlling ticks and tick-borne diseases by frequent applications of acaricides (e.g., dipping) is costly, and can leave treated livestock vulnerable to epizootics of tick-borne diseases should the system of applying acaricides break down. The concept of only applying acaricides on an infrequent (strategic) basis often relies on the target tick population displaying a seasonal cycle. However, as adult bont tick (Amblyomma hebraeum) infestations in Zimbabwe's lowveld do not have a strictly seasonal pattern of occurrence, it is recommended that tick control only be applied when bont tick infestations are equal to, or greater than, their economic damage threshold. The economic damage threshold is the minimum average weekly standard female tick burden sufficient to cause damage equal in dollar value to the costs of applying tick control. Assuming that each standard female tick represents a 10 gram weight loss, the economic damage threshold (standard female ticks/week) is equivalent to the ratio of the producer price of beef (liveweight equivalent): per head cost of dipping (Eqn (3)). To illustrate the application of the threshold methodology, it was assumed that the producer price of beef was Z$1.63/kg (U.S.$0.33/kg) and that tick control cost Z$0.29/hd/dip (U.S.$0.06/hd/dip). This gave a threshold of 18 standard female ticks/head/week. Using tick counts obtained from 20 Brahman cattle held at Mbizi in southeastern Zimbabwe, it was shown that for the 1988 calendar year there were only 32 weeks when the economic damage threshold was met or exceeded. This is substantially less that the 44 dippings per year that have been, until very recently, legally required in Zimbabwe. Sensitivity analysis showed that a 10% rise in the cost of dipping reduced to 23 (a 28% decrease) the number of weeks when tick burdens exceeded the economic damage threshold. By further assuming that an acaricide application and residual effects will cause a 3–5 week interval before the next application may be required, the number of weeks when the tick burden was equal to or greater than the threshold of 18 standard females/week fell to just 9–12 weeks. Three factors may cause an alteration in the economic damage threshold: i) tick burdens may cause damage to the udders; ii) secondary infestations (e.g., screw-worm) may cause economic damage; and iii) nutritional stress of the cattle may reduce the actual average per tick weight loss. Until further data becomes available, it is recommended that the economic damage threshold methodology be used as described here, and that farmers closely observe their herds during the suggested weekly tick sampling for udder damage and secondary infestations.     

The impact of natural infestations of ticks in Zambia on the productivity of cattle and implications for tick control strategies in central Africa

Studies were carried out in Zambia using a farming systems approach to quantify the effects of tick control on traditionally managed Sanga cattle. In the first trial to investigate the effect of tick control, significant improvements in liveweight gain (LWG) occurred only in periods of medium to high challenge with adult Amblyomma variegatum. However, during the first 18 months of the study when animals were young, the acaricide used caused a suppression in weight gains. A supplementary experimental study indicated that the adverse effects of ticks on weight gain may be delayed when animals are on a low plane of nutrition. In both studies on the effects of ticks on LWG, estimates of weight loss per female A. variegatum were in the range of 46 to 69 g. In the second long term trial to measure the impact of tick control on overall heard productivity, outputs of milk and weaner calf per livestock unit carrying capacity were about 25% higher in a tick-free herd. The annual cost of control in 1988 at ZK286.26/livestock unit was greater than the increase in value of the products at ZK175.48/livestock unit carrying capacity. It is concluded from these studies that intensive tick control in the multipurpose livestock system in Zambia is not justified in the absence of serious tick-borne disease. However, strategic seasonal tick control of adult ticks from November to April would probably be economically viable. Calves should not be treated until they are 3 months of age.     

Suppression of host-seeking Ixodes scapularis and Amblyomma americanum (Acari: Ixodidae) nymphs after dual applications of plant-derived acaricides in New Jersey

We evaluated the ability of dual applications of natural, plant-derived acaricides to suppress nymphal Ixodes scapularis Say and Amblyomma americanum (L.) (Acari: Ixodidae) in a Lyme disease endemic area of New Jersey. An aqueous formulation of 2% nootkatone provided >90% control of I. scapularis through 7 d. Control declined to 80.9% at 14 d, and a second application was made that provided >95% control through the remaining 4 wk of the nymphal season. Nootkatone provided >90% control of A. americanum through 35 d postapplication. Applications of 2% carvacrol and EcoTrol T&O resulted in rapid knockdown of both tick species, but control declined significantly to 76.7 and 73.7%, respectively, after 14 d when a second application was made that extended control of both tick species to between 86.2 and 94.8% at 21 d. Subsequently, control declined steadily in all plots by 42 d postapplication except for I. scapularis in carvacrol-treated plots, where levels of control >90% were observed through 35 d. Of the three compounds tested, 2% nootkatone provided the most consistent results, with 96.5 and 91.9% control of I. scapularis and A. americanum through 42 and 35 d, respectively. The ability of plant-derived natural products to quickly suppress and maintain significant control of populations of these medically important ticks may represent a future alternative to the use of conventional synthetic acaricides. In addition, the demonstrated efficacy of properly-timed backpack sprayer application may enable homeowner access to these minimal-risk acaricides.