Chemical attraction of Dermacentor variabilis ticks parasitic to Peromyscus leucopus based on host body mass and sex

Macroparasites are commonly aggregated on a small subset of a host population. Previous explanations for this aggregation relate to differences in immunocompetence or the degree to which hosts encounter parasites. We propose active tick host choice through chemical attraction as a potential mechanism leading to aggregated tick burdens. We test this hypothesis using a Y-maze olfactometer, comparing chemical attraction responses of larval and nymphal Dermacentor variabilis ticks parasitic to the white-footed mouse, Peromyscus leucopus, as a function of host sex and host body mass. We hypothesized that larger hosts and male hosts would be most attractive to searching ticks, as these hosts commonly have higher tick burdens in the field. Chemical attraction trials were run in the presence and absence of a known tick attractant, host-produced carbon dioxide (CO₂). Male hosts and larger hosts were preferred by nymphal D. variabilis in the presence and absence of CO₂, whereas larvae had no detectable host preference. The current study suggests that host-produced chemical cues may promote aggregated tick burdens among hosts of a single species based on host body mass and sex.     

Partitioning the aggregation of parasites on hosts into intrinsic and extrinsic components via an extended Poisson-gamma mixture model

It is well known that parasites are often highly aggregated on their hosts such that relatively few individuals host the large majority of parasites. When the parasites are vectors of infectious disease, a key consequence of this aggregation can be increased disease transmission rates. The cause of this aggregation, however, is much less clear, especially for parasites such as arthropod vectors, which generally spend only a short time on their hosts. Regression-based analyses of ticks on various hosts have focused almost exclusively on identifying the intrinsic host characteristics associated with large burdens, but these efforts have had mixed results; most host traits examined have some small influence, but none are key. An alternative approach, the Poisson-gamma mixture distribution, has often been used to describe aggregated parasite distributions in a range of host/macroparasite systems, but lacks a clear mechanistic basis. Here, we extend this framework by linking it to a general model of parasite accumulation. Then, focusing on blacklegged ticks (Ixodes scapularis) on mice (Peromyscus leucopus), we fit the extended model to the best currently available larval tick burden datasets via hierarchical Bayesian methods, and use it to explore the relative contributions of intrinsic and extrinsic factors on observed tick burdens. Our results suggest that simple bad luck-inhabiting a home range with high vector density-may play a much larger role in determining parasite burdens than is currently appreciated.     

Differential distribution of immature Ixodes dammini (Acari: Ixodidae) on rodent hosts

Ectoparasites such as ixodid ticks that remain attached to hosts for several days while feeding on blood are able to overcome the inflammatory and immune responses of some hosts and not others. The immature stages of the deer tick Ixodes dammini are found more frequently on the white-footed mouse, Peromyscus leucopus, than on other rodents. We propose that P. leucopus is more tolerant to I. dammini than is a less common host, the meadow vole, Microtus pennsylvanicus. To test this hypothesis, the distribution patterns and engorgement indices were determined for larval and nymphal I. dammini collected from wild-caught P. leucopus and M. pennsylvanicus. There were more immature ticks, which were more fully engorged, on P. leucopus than on M. pennsylvanicus. There were more and better engorged ticks on male than on female hosts. Laboratory studies on the number and weights of larval I. dammini collected off naive and previously exposed P. leucopus and M. pennsylvanicus support the results of the field study. Fewer larval ticks were recovered from previously exposed M. pennsylvanicus than P. leucopus, and the ticks weighed less. Larval and nymphal ticks aggregated among hosts in the study grid, and higher densities per male P. leucopus were correlated with higher engorgement indices, suggesting that immature I. dammini feed better at higher densities. The feeding success of I. dammini on its preferred host species might be due to its adaptation to the immune and inflammatory reactions of the host.     

Impact of the experimental removal of lizards on Lyme disease risk

The distribution of vector meals in the host community is an important element of understanding and predicting vector-borne disease risk. Lizards (such as the western fence lizard; Sceloporus occidentalis) play a unique role in Lyme disease ecology in the far-western United States. Lizards rather than mammals serve as the blood meal hosts for a large fraction of larval and nymphal western black-legged ticks (Ixodes pacificus--the vector for Lyme disease in that region) but are not competent reservoirs for the pathogen, Borrelia burgdorferi. Prior studies have suggested that the net effect of lizards is to reduce risk of human exposure to Lyme disease, a hypothesis that we tested experimentally. Following experimental removal of lizards, we documented incomplete host switching by larval ticks (5.19%) from lizards to other hosts. Larval tick burdens increased on woodrats, a competent reservoir, but not on deer mice, a less competent pathogen reservoir. However, most larvae failed to find an alternate host. This resulted in significantly lower densities of nymphal ticks the following year. Unexpectedly, the removal of reservoir-incompetent lizards did not cause an increase in nymphal tick infection prevalence. The net result of lizard removal was a decrease in the density of infected nymphal ticks, and therefore a decreased risk to humans of Lyme disease. Our results indicate that an incompetent reservoir for a pathogen may, in fact, increase disease risk through the maintenance of higher vector density and therefore, higher density of infected vectors.     

Differential detachment from resting hosts of replete larval and nymphal Ixodes ticks

To determine whether replete subadult Ixodes ticks detach more frequently from resting than from active hosts, diverse rodents and lizards were caged in an apparatus designed to record the ticks' sites of detachment relative to the resting site of the host. Replete larval Ixodes ricinus and Ixodes dammini accumulated mainly beneath the resting places of the mice (Apodemus agrarius and Peromyscus leucopus) most frequently parasitized in nature. Although nymphal I. ricinus similarly detached where these mice rested, nymphal I. dammini detached more randomly. When lizards were used as hosts, both subadult stages of I. ricinus tended to detach away from their main resting sites; these ticks detached from squirrels more randomly. Detachment ratios for other rodent hosts, that are abundantly infested by the larvae of these ticks in nature (Apodemus flavicollis and Clethrionomys glareolus), could not be derived because nymphs generally failed to attach. Our observations are consistent with reports that both subadult stages of I. dammini, but not the adult, feed on the same kind of nest-dwelling hosts and that the host range of I. ricinus is less focused. Detachment of mouse-feeding larvae from resting mice promotes subsequent nymphal attachment to conspecific hosts, and the absence of such behavior among nymphs facilitates access of the resulting adults to deer.     

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.     

Predicting larval tick burden on white-footed mice with an artificial neural network

White-footed mice are important hosts for immature blacklegged ticks (Ixodes scapularis) and the most competent reservoir hosts for several tick-borne pathogens, including the agent of Lyme disease, in eastern North America. The distribution of larval ticks on individual mice tends to be highly heterogeneous, potentially resulting in few individual hosts causing the majority of host-to-tick transmission events. In this study, we created an artificial neural network (ANN) model using a 20 year data set from Millbrook, NY, to understand which attributes of mice or the environment predict high larval burden. Furthermore, we performed a sensitivity analysis to explore the importance of, and interactions between, the most influential attributes. Our analysis indicated that highest larval burden is predicted in warmer and drier than average years when host abundance is low, and that climatic conditions and host density are far more important in predicting larval burden than traits of individual mice, a finding that could have human health implications within the context of a warming climate. Practically, our results suggest that instead of basing tick-control treatments on particular attributes of hosts, treatments should be targeted based on climate factors. Additionally, our results highlight the importance of including variable interactions in models aiming to predict vector (tick) aggregation, and, most broadly, demonstrate the utility of ANNs in understanding aggregation of ticks and other vectors.     

Characteristics of Garden Dormice That Contribute to Their Capacity as Reservoirs for Lyme Disease Spirochetes

To describe the contribution of garden dormice to the epizootiology of Lyme disease, we compared their reservoir capacity for these pathogens to that of other sympatric hosts. Garden dormice are trapped most abundantly during early spring and again during midsummer, when their offspring forage. They are closely associated with moist forests. Garden dormice serve as hosts to nymphal ticks far more frequently than do other small mammals. Spirochetal infection is most prevalent in dormice, and many more larval ticks acquire infection in the course of feeding on these than on other rodents in the study site. Mature dormice appear to contribute more infections to the vector population than juveniles do. Replete larval ticks generally detach while their dormouse hosts remain within their nests. The population of garden dormice contributes five- to sevenfold more infections to the vector population than the mouse population does. Their competence, nymphal feeding density, and preference for a tick-permissive habitat combine to favor garden dormice over other putative reservoir hosts of Lyme disease spirochetes.     

Empirical evidence for key hosts in persistence of a tick-borne disease

An important epidemiological consequence of aggregated host-parasite associations occurs when parasites are vectors of pathogens. Those hosts that attract many vectors will tend to be the focus of transmission. But to what extent, and can we identify characteristics of these key hosts? We investigated these questions with respect to the host-tick relationship of the yellow-necked mouse, Apodemus flavicollis, a critical host in the maintenance of the zoonotic disease, tick-borne encephalitis. Transmission of the virus occurs when ticks feed in a 'co-feeding' aggregation. Thus, the number and frequency of co-feeding groups provides an estimate of the potential rate of virus transmission. We recorded the spatio-temporal variations in co-feeding on a population of rodents in conjunction with recording individual host characteristics. Using Lorenz curves, we revealed conformation of tick-borne encephalitis transmission potential to the 20/80 Rule, where the 20% of hosts most infested with ticks were accountable for 80% of transmission potential. Hosts in the transmission cohort were identified as the sexually mature males of high body mass. Therefore control efforts targeted at this group would substantially reduce transmission potential compared to non-targeted control of the population, which resulted in a linear reduction in transmission potential. Focusing on the 'wrong' functional group would have little impact upon transmission potential until a considerable proportion of the population had been subject to control. However, individuals can change their functional status over time making it difficult to predict the contribution of these individuals to future transmission.     

Life cycle and host specificity of Amblyomma triste (Acari: Ixodidae) under laboratory conditions

We report biological data of two generations of Amblyomma triste in laboratory and compared the suitability of different host species. Infestations by larval and nymphal stages were performed on guinea pigs (Cavia porcellus), chickens (Gallus gallus), rats (Rattus norvegicus), rabbits (Oryctolagus cuniculus), wild mice (Calomys callosus), dogs (Canis familiaris) and capybaras (Hydrochaeris hydrochaeris). Infestations by adult ticks were performed on dogs, capybaras and rabbits. Tick developmental periods were observed in an incubator at 27 degrees C and RH 90%. Guinea pigs were the most suitable hosts for larvae and nymphs, followed by chickens. The remaining host species were less suitable for immature ticks as fewer engorged ticks were recovered from them. Mean larval feeding periods varied from 3.8 to 4.7 d between different host species. Mean larval premolt periods ranged from 8.9 to 10.4 d. Nymphal mean feeding periods varied from 4.2 to 6.2 d for ticks fed on different host species. Premolt period of male nymphs (mean: 15.4 d) was significantly longer than that of female nymphs (14.7 d). Female nymphs were significantly heavier than male nymphs. The overall sex ratio of the adult ticks emerged from nymphs was 0.9:1 (M:F). Capybaras were the most suitable host for the tick adult stage as significantly more engorged females were recovered from them and these females were significantly heavier than those recovered from dogs or rabbits. The life cycle of A. triste in laboratory could be completed in an average period of 155 d. The potential role of guinea pigs, birds and capybaras, as hosts for A. triste in nature, is discussed.     

A molecular marker for the identification of the zoonotic reservoirs of Lyme borreliosis by analysis of the blood meal in its European vector Ixodes ricinus.

The efficacy of the mitochondrially encoded cytochrome b gene as a molecular marker for the discrimination of the reservoir host species of the Lyme borreliosis spirochete, Borrelia burgdorferi sensu lato (s.l.), in its European vector Ixodes ricinus (Acari: Ixodidae) was determined. Degenerate PCR primers were designed which amplified orthologous regions of the cytochrome b gene in several animal species which act as B. burgdorferi s.l. reservoirs and hosts for I. ricinus. PCR products were amplified and characterized by hybridization and restriction fragment length polymorphism analysis. Restriction fragment length polymorphism analysis of a 638-bp PCR product with HaeIII and DdeI revealed unique restriction fragment profiles, which allowed the taxonomic identification of animals to the genus level. A system was devised for the detection of the larval host blood meal from the remnants in unfed nymphal I. ricinus ticks by nested PCR amplification. An inverse correlation was demonstrated between amplicon size and successful PCR amplification of host DNA from the nymphal stage of the tick. The stability of the cytochrome b product as a marker for the identification of the larval host species in the nymphal instar was demonstrated up to 200 days after larval ingestion (approximately 165 days after molting) by reverse line blotting with a host-specific probe. This assay has the potential for the determination of the reservoir hosts of B. burgdorferi s.l. by using extracts from the same individual ticks for both the identification of the host species and the detection of the Lyme borreliosis spirochete.     

Effects of a zoonotic pathogen,Borrelia burgdorferi,on the behavior of a key reservoir host

Most emerging infectious diseases of humans are transmitted to humans from other animals. The transmission of these “zoonotic” pathogens is affected by the abundance and behavior of their wildlife hosts. However, the effects of infection with zoonotic pathogens on behavior of wildlife hosts, particularly those that might propagate through ecological communities, are not well understood. Borrelia burgdorferi is a bacterium that causes Lyme disease, the most common vector‐borne disease in the USA and Europe. In its North American range, the pathogen is most frequently transmitted among hosts through the bite of infected blacklegged ticks (Ixodes scapularis). Using sham and true vaccines, we experimentally manipulated infection load with this zoonotic pathogen in its most competent wildlife reservoir host, the white‐footed mouse, Peromyscus leucopus, and quantified the effects of infection on mouse foraging behavior, as well as levels of mouse infestation with ticks. Mice treated with the true vaccine had 20% fewer larval blacklegged ticks infesting them compared to mice treated with the sham vaccine, a significant difference. We observed a nonsignificant trend for mice treated with the true vaccine to be more likely to visit experimental foraging trays (20%–30% effect size) and to prey on gypsy moth pupae (5%–20% effect size) compared to mice treated with the sham vaccine. We observed no difference between mice on true‐ versus sham‐vaccinated grids in risk‐averse foraging. Infection with this zoonotic pathogen appears to elicit behavioral changes that might reduce self‐grooming, but other behaviors were affected subtly or not at all. High titers of B. burgdorferi in mice could elicit a self‐reinforcing feedback loop in which reduced grooming increases tick burdens and hence exposure to tick‐borne pathogens.     

Real-time PCR for simultaneous detection and quantification of Borrelia burgdorferi in field-collected Ixodes scapularis ticks from the Northeastern United States

The density of spirochetes in field-collected or experimentally infected ticks is estimated mainly by assays based on microscopy. In this study, a real-time quantitative PCR (qPCR) protocol targeting the Borrelia burgdorferi-specific recA gene was adapted for use with a Lightcycler for rapid detection and quantification of the Lyme disease spirochete, B. burgdorferi, in field-collected Ixodes scapularis ticks. The sensitivity of qPCR for detection of B. burgdorferi DNA in infected ticks was comparable to that of a well-established nested PCR targeting the 16S-23S rRNA spacer. Of the 498 I. scapularis ticks collected from four northeastern states (Rhode Island, Connecticut, New York, and New Jersey), 91 of 438 (20.7%) nymphal ticks and 15 of 60 (25.0%) adult ticks were positive by qPCR assay. The number of spirochetes in individual ticks varied from 25 to 197,200 with a mean of 1,964 spirochetes per nymphal tick and a mean of 5,351 spirochetes per adult tick. No significant differences were found in the mean numbers of spirochetes counted either in nymphal ticks collected at different locations in these four states (P = 0.23 by one-way analysis of variance test) or in ticks infected with the three distinct ribosomal spacer restriction fragment length polymorphism types of B. burgdorferi (P = 0.39). A high degree of spirochete aggregation among infected ticks (variance-to-mean ratio of 24,877; moment estimate of k = 0.279) was observed. From the frequency distribution data and previously published transmission studies, we estimated that a minimum of 300 organisms may be required in a host-seeking nymphal tick to be able to transmit infection to mice while feeding on mice. These data indicate that real-time qPCR is a reliable approach for simultaneous detection and quantification of B. burgdorferi infection in field-collected ticks and can be used for ecological and epidemiological surveillance of Lyme disease spirochetes.     

Life cycle of Amblyomma cooperi (Acari: Ixodidae) using capybaras (Hydrochaeris hydrochaeris) as hosts

The life cycle of Amblyomma cooperi was evaluated under laboratory conditions testing different host species. Larval infestations were performed on chickens (Gallus gallus) and capybaras (Hydrochaeris hydrochaeris). Nymphal infestations were performed on G. gallus, H. hydrochaeris, guinea pigs (Cavia porcellus) and wild mice (Calomys callosus). Infestations by adult ticks were performed only on capybaras. All free-living stages were observed in darkness at 27 degrees C and RH 85%. Capybaras were significantly (p < 0.05) the most suitable hosts for immature ticks, with the highest larval (63.6%) and nymphal (48%) recovery. Larval and nymphal feeding and premolt periods were significantly different (p < 0.05) between ticks fed on different host species. Male nymphs showed premolt period significantly shorter (p < 0.05) than female nymphs. The overall sex ratio of adult ticks was 0.92:1 (M:F). Infestations by adult ticks on capybaras yielded more than 76% of engorged female recovery. Only three out of 33 engorged females fed on capybaras did not lay fertile eggs. The life cycle of A. cooperi in laboratory, reported for the first time, was completed in an average period of 189.4 days. During the premolt period, all A. cooperi engorged nymphs secreted distinct blackish drops, which seem to be inherent to this species. Our results, associated data in the literature, confirm the high suitability of capybaras for the adult stage of A. cooperi and also indicate this animal species as a primary host for immature stages of A. cooperi in nature. On the other hand, the results of larval and nymphal infestation on chickens and guinea pigs suggest that birds and wild guinea pigs, which are also present in the distribution area of A. cooperi in South America, could be potentially infested by A. cooperi immature stages in nature.     

Borrelia Burgdorferi Sensu Lato in Ixodes Ricinus Ticks and Rodents in a Recreational Park in South-Western Ireland

Ixodes ricinus ticks infected with Borrelia burgdorferi sensu lato were numerous on the edges of paths and roads in a recreational park in south-western Ireland. The abundance of ticks at different sites was related to the presence of deer, but a negative relationship was shown between tick abundance and tick infection rates. This is thought to be due to the deposition of large numbers of uninfected ticks by deer, which are apparently not good reservoir hosts of B. burgdorferi s.l. Blood meal analysis only detected deer DNA in uninfected nymphs. Reservoir competent rodents, Apodemus sylvaticus and Clethrionomys glareolus, were abundant at all sites and a high proportion of captured specimens were infested with larval ticks. However, very few rodents were infected with B. burgdorferi s.l. and none of the unfed infected nymphs analysed for the identity of their larval blood meal had fed on rodents. The spirochaetes detected in I. ricinus in the study area may be poorly adapted to rodents or are not transmitted readily because of the absence of nymphal infestation. The majority of spirochaetes in these ticks were apparently acquired from non-rodent hosts, such as birds.     

Vectorial capacity of North American Ixodes ticks.

Ixodes dammini, the vector of Lyme disease and babesiosis, is distributed in various locations in the northeastern quadrant of the United States and nearby Canada. The life cycle of this tick, which includes larval, nymphal, and adult stages, spans at least two years. The tick over-winters between larval and nymphal feeding. Horizontal transmission of pathogens is facilitated by a feeding pattern in which both the larval and nymphal stages feed on the white-footed mouse, Peromyscus leucopus, and by a seasonal pattern of activity in which nymphs precede larvae. The species range appears to have expanded from a single island location, and has invaded new sites since the 1940s, some as recently as 1980. This increased abundance appears to be related to the increased abundance of deer, the preferred host of the adult stage. I. muris predominated in coastal Massachusetts before I. dammini became abundant, but is probably now extinct. I. scapularis, which is present in the southern U.S., is a poor vector of mouse parasites because about 90 percent of these immature ticks feed on lizards. To the extent that horizontal transmission occurs, we suggest that mice serve as the principal reservoir for the Lyme spirochete as well as Babesia microti.     

Effect of host lizard anemia on host choice and feeding rate of larval western black-legged ticks (Ixodes pacificus)

Although ticks are known to exhibit preferences among host species, there is little evidence that ticks select hosts within a species based on physiological condition. It may be beneficial for ticks to choose hosts that are easier to feed upon if the ticks can perceive indicative chemical or other signals from the host. For example, if ticks can detect host hematocrit they may choose hosts with high hematocrit, facilitating a faster blood meal. It may similarly be adaptive for ticks to avoid anemic hosts because it may be difficult for them to obtain an adequate meal and feeding duration may be extended. We tested the hypothesis that larval western black-legged ticks (Ixodes pacificus) detect host hematocrit using external cues and choose healthy over anemic hosts, allowing them to feed more quickly. We presented groups of larval ticks with pairs of healthy and anemic male western fence lizards (Sceloporus occidentalis), allowed them to select a host, and measured the feeding duration of the ticks. We found that the ticks did not exhibit a statistically significant preference for healthy over anemic lizards, but that the ticks fed to repletion significantly faster on healthy hosts than on anemic hosts. Larval ticks may not be able to detect external cues indicating the health of the host, at least not in terms of their hematocrit. The extended feeding duration likely reflects the extra time needed for the ticks to concentrate the blood meal of their anemic hosts.     

Expression of acquired immunity to immature stages of the tick Rhipicephalus evertsi evertsi by rabbits and guinea-pigs

Acquired immunity in guinea-pigs and rabbits to immature stages of the two-host tick Rhipicephalus evertsi evertsi Neumann was demonstrated. Repeated infestations of both hosts with larvae resulted in a significant reduction in the weight of later engorged nymphs. A sharp decline in the numbers of nymphs which successfully fed on both hosts was also observed. This study provides evidence for a gradual decrease in the mean weight of engorged nymphs towards the end of the detachment period suggesting that, in two-host ticks, the onset of nymphal feeding acts as an immune booster in a host already primed by the larval feed and that this results in a reduced feeding performance.     

Life cycle and host specificity of Amblyomma triste (Acari: Ixodidae) under laboratory conditions

We report biological data of two generations of Amblyomma triste in laboratory and compared the suitability of different host species. Infestations by larval and nymphal stages were performed on guinea pigs (Cavia porcellus), chickens (Gallus gallus), rats (Rattus norvegicus), rabbits (Oryctolagus cuniculus), wild mice (Calomys callosus), dogs (Canis familiaris) and capybaras (Hydrochaeris hydrochaeris). Infestations by adult ticks were performed on dogs, capybaras and rabbits. Tick developmental periods were observed in an incubator at 27 °C and RH 90%. Guinea pigs were the most suitable hosts for larvae and nymphs, followed by chickens. The remaining host species were less suitable for immature ticks as fewer engorged ticks were recovered from them. Mean larval feeding periods varied from 3.8 to 4.7 d between different host species. Mean larval premolt periods ranged from 8.9 to 10.4 d. Nymphal mean feeding periods varied from 4.2 to 6.2 d for ticks fed on different host species. Premolt period of male nymphs (mean: 15.4 d) was significantly longer than that of female nymphs (14.7 d). Female nymphs were significantly heavier than male nymphs. The overall sex ratio of the adult ticks emerged from nymphs was 0.9:1 (M:F). Capybaras were the most suitable host for the tick adult stage as significantly more engorged females were recovered from them and these females were significantly heavier than those recovered from dogs or rabbits. The life cycle of A. triste in laboratory could be completed in an average period of 155 d. The potential role of guinea pigs, birds and capybaras, as hosts for A. triste in nature, is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Interspecific variations in attachment sites and density assessment in femaleIxodes rubicundus (Acari: Ixodidae) on domestic and natural hosts

The attachment sites of female Karoo paralysis ticks (Ixodes rubicundus) and equations to assess tick burdens on domestic (sheep and goats) and natural (mountain reedbuck) hosts were determined. No intraspecific differences in attachment sites for Merino sheep and Angora goats could be observed, but interspecific differences were evident.Ixodes rubicundus in general attaches on the ventral aspects of the host. More than 50% of these ticks attached to the fron quarters of the body of the domestic host, and 60% to the hindquarters of the natural host. The length of Merino sheep wool had no affect on either the attachment site or number of attached ticks. In general, more ticks were found attached to the woollen than non-woollen body areas. Equations are provided which can estimate tick burdens on hosts by counting the ticks on two or three selected body areas of the host. To facilitate sampling procedures, approximate sample sizes for the various hosts needed to estimate average tick burdens have been provided. Depending on the level of precision required (with bounds on the error of estimation equal to 1, 2 or 5 ticks), the number of animals in a flock of 200 Angora goats that must be examined varied from 26 to 85.