Systemic efficacy of nodulisporamides against fleas on dogs.

Nodulisporic acid A (NSA) has been shown previously to be safe in dogs and to deliver >90% flea control for 4 days following a single oral administration. Three newly prepared nodulisporamide derivatives were subsequently identified from an artificial membrane flea feeding system as exhibiting potency substantially greater than NSA. To determine if they have superior in vivo activity, these 3 nodulisporamides, as well as NSA, were evaluated in dogs at 15 mg/kg/os. Parasite challenges were made by placing 100 live Ctenocephalides felis fleas onto the dorsum of dogs every 48 hr and examining efficacy at each of those intervals over a 22-day period. Results showed that NSA produced >90% efficacy at day 2 and 81% efficacy at day 4, and its residual flea killing fell to approximately 50% by day 6 posttreatment. All dogs treated with the 3 new experimental nodulisporamides were 100% protected from flea challenges to day 8 posttreatment, and 2 of the compounds continued to produce >90% residual activity to 2 wk posttreatment. Pharmacokinetic analysis showed that plasma profiles and half-lives of NSA and these 3 new compounds correlated closely with flea efficacy. These results demonstrate that specific substitutions to the pharmacophore of NSA can substantially increase the duration of activity against fleas.     

First blood meal of Ctenocephalides canis (Siphonaptera: Pulicidae) on dogs: time to initiation of feeding and duration

Two experiments were conducted on dogs to evaluate interval to initiation and duration of the first blood meal of Ctenocephalides canis (Curtis). Percentage of fed male and female fleas was calculated for fleas held on dogs for 5, 15, 30, 60 min, 6, and 24 hr. Duration of first blood meal was also measured for individual fleas confined on dogs. When fleas were free in the hair coat, 21.2% had begun blood feeding within 5 min. After 1 hr, 72.5% of fleas had fed. After 6 hr, 95.2% of males and 100% of females had taken a blood meal, and 24 hr after deposition all fleas had fed. There was no significant difference between the 2 sexes. The mean delay between deposition and biting for fleas that began feeding within 15 min was 2 min 52 sec +/- 3 min 2 sec for female fleas and 3 min 8 sec +/- 2 min 45 sec for males. The mean duration of female and male meals was 5 min 3 sec +/- 3 min 41 sec and 6 min 9 sec +/- 6 min 8 sec, respectively. There was no significant difference between the 2 sexes. The dog flea took its blood meal on dogs more slowly than the cat flea did on cats; this meal was significantly longer for Ctenocephalides felis felis (Bouche) than for C. canis.     

Comparative efficacy on dogs of a single topical treatment with fipronil/(S)-methoprene or weekly physiological hygiene shampoos against Ctenocephalides felis in a simulated flea-infested environment

Flea infestations of pets continue to persist due to the lack of knowledge of flea biology and ecology. It is not unusual that pet owners believe regular hygiene, such as shampooing their dogs can replace regular insecticidal treatment. The objective of this study was to compare in a flea simulated environment, modelling exposure similar to that found in a home, that the use of regular physiological shampoo does not control fleas adequately when compared to a long acting topical formulation. Three groups of six dogs were formed: one untreated control group, one group treated monthly with the topical formulation of fipronil/(S)-methoprene, and a third group treated weekly with a hygiene shampoo. All dogs were infested with adult unfed Ctenocephalides felis fleas (200±5) on Days -28 and -21. Each animal's sleeping box was fitted with a plastic cup mounted to the inside roof of the box. The sleeping bench of each animal was covered with a carpet to accommodate flea development. The dogs were maintained in their kennels throughout the study. In order to maintain the environmental flea challenge, C. felis pupae (100±5) were placed in the plastic cup in each animal's sleeping box on Days -14, -7, 0, 7, 14, 21, 28, 35 and 42. The dogs were combed and fleas counted weekly on Days -1, 3, 10, 17, 24, 31, 38, 45, and 51. The fleas were placed immediately back on the dogs. On Day 60, fleas were counted and removed. Flea infestations in the untreated control group at each count averaged between 46.2 and 74.2 fleas throughout the study. The average number of fleas infesting dogs was significantly different (p<0.05) between the untreated and the two treatment groups and between the two treatment groups at all counts throughout the two months study (Days 3, 10, 17, 24, 31, 38, 45, 51 and 60). The efficacy was never below 99.1% in the fipronil/(S)-methoprene group, and efficacy in the shampoo group was never above 79.2%. Weekly shampooing in treatment group 3 was intentionally delayed after Day 42, to evaluate wether missing a weekly bath would affect the flea population. The Day 48 data indicate that forgetting or delaying a single weekly shampooing resulted in a clear increase in flea numbers and a significant decrease in efficacy from 68.2% to 34.8%. The fipronil(S)/methoprene treatment allowed a continuous control as demonstrated by the high efficacy against fleas, and also the number of flea-free dogs on seven of the nine weekly assessments, in spite of what was essentially a continuous flea challenge model.     

Prevalence of flea infestation in dogs and cats in Hungary combined with a survey of owner awareness

A survey was conducted in order to gain current information on flea species (Siphonaptera: Pulicidae) infesting dogs and cats living in urban and rural areas of Hungary, along with data on the factors that affect the presence, distribution and seasonality of infestation. In addition, owner awareness of flea infestation was evaluated. Practitioners in 13 veterinary clinics were asked to examine all dogs and cats attending the clinic and to collect fleas, when present, on 2 days in each month from December 2005 to November 2006. They also completed a questionnaire for each animal examined. A total of 319 dogs (14.1%) were found to be infested; the highest prevalence (27.1%) of infestation on dogs occurred in August and the lowest (5.4%) in May. Prevalence of fleas on cats was higher (22.9%); the highest (35.0%) and lowest (8.1%) prevalences occurred in July and April, respectively. Fleas were more prevalent in rural (387/1924 animals, 20.2%) than in urban (161/1343 animals, 12.0%) areas. Three species, Ctenocephalides felis (Bouché), Ctenocephalides canis (Curtis) and Pulex irritans L., were found. On dogs, the prevalence of C. canis alone was 53.0%, whereas that of C. felis alone was 36.0%. Only 19 specimens of P. irritans were found on 14 dogs from rural habitats only. Prevalence of C. felis only on cats was 94.3%; the remaining cats were infested with either C. canis or with mixed infestations of C. felis and C. canis. More than half (51.4%) of the owners of infested dogs and cats had not used flea control products in the past year or more, and five times as many owners in rural than urban areas had not used flea control products in the same period. Very few owners reported having attempted to kill fleas in their animals' environment; instead, they believed that fleas were acquired from other cats or dogs.     

Comparative efficacy on dogs of a single topical treatment with the pioneer fipronil/(S)-methoprene and an oral treatment with spinosad against Ctenocephalides felis

In the study reported here, the pioneer fipronil/(S)-methoprene topical product (FRONTLINE^® PLUS, Merial Limited, Duluth, GA) was compared to the oral spinosad product (COMFORTIS^® Elanco, Greenfield, IN) for efficacy against adult fleas and preventing egg production. The product presentations, doses and labelling were the one applicable in the USA. Using a standard protocol, 200 cat fleas of mixed sex were applied to dogs on Days 1, 7, 14, 21, 28, 35, and 42. Dogs were combed to remove fleas 24 hours post-infestation, the fleas were counted, collected, and then reapplied to each dog following completion of their respective count. At 48 hours post-infestation, comb counts were performed and fleas were removed. No fleas were collected from any dog in the fipronil/(S)-methoprene group at any 24 or 48 hours post-infestation assessment throughout the six weeks study, yielding a preventive efficacy of 100%. For the spinosad treatment, efficacy was 100% at 24 hours and 48 hours through Day 16, and thereafter declined. The results observed in the spinosad-treated dogs were highly variable between animals. At the 24 and 48 hours counts following the Day 21 infestation, only five of eight spinosad-treated dogs (62.5%) were flea-free. Following the Day 28 infestation, spinosad efficacy fell to 85% and 89%, for the 24 hours and 48 hours counts, and only two dogs (25%) were flea free, compared to 100% flea-free dogs in the fipronil/(S)-methoprene group. No fleas were collected from the fipronil/(S)- methoprene treated dogs throughout the entire study, therefore, no eggs were collected at any time from any dog in the group. However, in the spinosad group adult fleas were found on dogs starting on Day 21 and by Day 30, 42 eggs were collected from one dog that had 107 adult fleas counted at 48 hours. At Day 37 and Day 49, more than 100 eggs were collected from each dog in the spinosad-treated and control groups.     

Horizontal transmission of Rickettsia felis between cat fleas, Ctenocephalides felis

Rickettsia felis is a rickettsial pathogen primarily associated with the cat flea, Ctenocephalides felis. Although laboratory studies have confirmed that R. felis is maintained by transstadial and transovarial transmission in C. felis, distinct mechanisms of horizontal transmission of R. felis among cat fleas are undefined. Based on the inefficient vertical transmission of R. felis by cat fleas and the detection of R. felis in a variety of haematophagous arthropods, we hypothesize that R. felis is horizontally transmitted between cat fleas. Towards testing this hypothesis, flea transmission of R. felis via a bloodmeal was assessed weekly for 4 weeks. Rhodamine B was used to distinguish uninfected recipient and R. felis-infected donor fleas in a rickettsial horizontal transmission bioassay, and quantitative real-time PCR assay was used to measure transmission frequency; immunofluorescence assay also confirmed transmission. Female fleas acquired R. felis infection more readily than male fleas after feeding on a R. felis-infected bloodmeal for 24 h (69.3% and 43.3%, respectively) and both Rickettsia-uninfected recipient male and female fleas became infected with R. felis after cofeeding with R. felis-infected donor fleas (3.3-40.0%). Distinct bioassays were developed to further determine that R. felis was transmitted from R. felis-infected to uninfected fleas during cofeeding and copulation. Vertical transmission of R. felis by infected fleas was not demonstrated in this study. The demonstration of horizontal transmission of R. felis between cat fleas has broad implications for the ecology of R. felis rickettsiosis.     

Rickettsia infection in five areas of the state of São Paulo, Brazil

This study investigated rickettsial infection in animals, humans, ticks, and fleas collected in five areas of the state of S?o Paulo. Eight flea species (Adoratopsylla antiquorum antiquorum, Ctenocephalides felis felis, Polygenis atopus, Polygenis rimatus, Polygenis roberti roberti, Polygenis tripus, Rhopalopsyllus lugubris, and Rhopalopsyllus lutzi lutzi), and five tick species (Amblyomma aureolatum, Amblyomma cajennense, Amblyomma dubitatum, Ixodes loricatus, and Rhipicephalus sanguineus) were collected from dogs, cats, and opossums. Rickettsia felis was the only rickettsia found infecting fleas, whereas Rickettsia bellii was the only agent infecting ticks, but no animal or human blood was shown to contain rickettsial DNA. Testing animal and human sera by indirect immunofluorescence assay against four rickettsia antigens (R. rickettsii, R. parkeri, R. felis, and R. bellii), some opossum, dog, horse, and human sera reacted to R. rickettsii with titers at least four-fold higher than to the other three rickettsial antigens. These sera were considered to have a predominant antibody response to R. rickettsii. Using the same criteria, opossum, dog, and horse sera showed predominant antibody response to R. parkeri or a very closely related genotype. Our serological results suggest that both R. rickettsii and R. parkeri infected animals and/or humans in the studied areas.     

Wolbachia endosymbionts in fleas (Siphonaptera)

Intracellular endosymbionts, Wolbachia spp., have been reported in many different orders of insects and in nematodes but not previously in fleas. This is the first conclusive report of Wolbachia spp. within members of the Siphonaptera. Using nested polymerase chain reaction (PCR) targeting of the 16S ribosomal RNA gene, we screened for Wolbachia spp. in fleas collected from 3 counties in Georgia and 1 in New York. The prevalence of Wolbachia spp. detected varied among the 6 different species screened: 21% in the cat flea Ctenocephalides felis (n = 604), 7% in the dog flea C. canis (n = 28), 25% in Polygenus gwyni (n = 8), 80% in Orchopeas howardi (n = 15), 94% in Pulex simulans (n = 255), and 24% in the sticktight flea Echidnophaga gallinacea (n = 101). Wolbachia spp. infection in fleas was confirmed by sequencing positive PCR products, comparing sequenced 16S ribosomal DNA (rDNA) with Wolbachia spp. sequences in GenBank using BLAST search, and subjecting sequence data to phylogenetic analysis. For further confirmation, 16S rDNA-positive samples were reamplified using the wsp gene.     

Prairie dog presence affects occurrence patterns of disease vectors on small mammals

Wildlife disease is recognized as a burgeoning threat to imperiled species and aspects of host and vector community ecology have been shown to have significant effects on disease dynamics. The black‐tailed prairie dog is a species of conservation concern that is highly susceptible to plague, a flea‐transmitted disease. Prairie dogs (Cynomys) alter the grassland communities in which they exist and have been shown to affect populations of small rodents, which are purported disease reservoirs. To explore potential ecological effects of black‐tailed prairie dogs on plague dynamics, we quantified flea occurrence patterns on small mammals in the presence and absence of prairie dogs at 8 study areas across their geographic range. Small mammals sampled from prairie dog colonies showed significantly higher flea prevalence, flea abundance, and relative flea species richness than those sampled from off‐colony sites. Successful plague transmission likely is dependent on high prevalence and abundance of fleas that can serve as competent vectors. Prairie dogs may therefore facilitate the maintenance of plague by increasing flea occurrence on potential plague reservoir species. Our data demonstrate the previously unreported ecological influence of prairie dogs on vector species assemblages, which could influence disease dynamics.     

Potential Effects of Environmental Conditions on Prairie Dog Flea Development and Implications for Sylvatic Plague Epizootics

Fleas are common ectoparasites of vertebrates worldwide and vectors of many pathogens causing disease, such as sylvatic plague in prairie dog colonies. Development of fleas is regulated by environmental conditions, especially temperature and relative humidity. Development rates are typically slower at low temperatures and faster at high temperatures, which are bounded by lower and upper thresholds where development is reduced. Prairie dogs and their associated fleas (mostly Oropsylla spp) live in burrows that moderate outside environmental conditions, remaining cooler in summer and warmer in winter. We found burrow microclimates were characterized by stable daily temperatures and high relative humidity, with temperatures increasing from spring through summer. We previously showed temperature increases corresponded with increasing off-host flea abundance. To evaluate how changes in temperature could affect future prairie dog flea development and abundance, we used development rates of O. montana (a species related to prairie dog fleas), determined how prairie dog burrow microclimates are affected by ambient weather, and combined these results to develop a predictive model. Our model predicts burrow temperatures and flea development rates will increase during the twenty-first century, potentially leading to higher flea abundance and an increased probability of plague epizootics if Y. pestis is present.

     

Local factors associated with on‐host flea distributions on prairie dog colonies

Outbreaks of plague, a flea‐vectored bacterial disease, occur periodically in prairie dog populations in the western United States. In order to understand the conditions that are conducive to plague outbreaks and potentially predict spatial and temporal variations in risk, it is important to understand the factors associated with flea abundance and distribution that may lead to plague outbreaks. We collected and identified 20,041 fleas from 6,542 individual prairie dogs of four different species over a 4‐year period along a latitudinal gradient from Texas to Montana. We assessed local climate and other factors associated with flea prevalence and abundance, as well as the incidence of plague outbreaks. Oropsylla hirsuta, a prairie dog specialist flea, and Pulex simulans, a generalist flea species, were the most common fleas found on our pairs. High elevation pairs in Wyoming and Utah had distinct flea communities compared with the rest of the study pairs. The incidence of prairie dogs with Yersinia pestis detections in fleas was low (n = 64 prairie dogs with positive fleas out of 5,024 samples from 4,218 individual prairie dogs). The results of our regression models indicate that many factors are associated with the presence of fleas. In general, flea abundance (number of fleas on hosts) is higher during plague outbreaks, lower when prairie dogs are more abundant, and reaches peak levels when climate and weather variables are at intermediate levels. Changing climate conditions will likely affect aspects of both flea and host communities, including population densities and species composition, which may lead to changes in plague dynamics. Our results support the hypothesis that local conditions, including host, vector, and environmental factors, influence the likelihood of plague outbreaks, and that predicting changes to plague dynamics under climate change scenarios will have to consider both host and vector responses to local factors.     

Notes on fleas (Siphonaptera) in plague foci on the Tay Nguyen plateau (Vietnam)]

The fleas of wild and commensal small mammals, domestic animals (dogs, cats) and free-living flea forms in houses have been collected in plague nidi of Tay Nguyen plate, Dak-Lak province, Vietnam. Pulex irritans, Ctenocephalides felis felis, Ct. felis orientis were found in the houses on dogs, cats and on the ground floor. Commensal rats in populated areas were infested by Xenopsylla cheopis and rarely by Lentistivalius klossi. The agricultural zone was inhabited by both home and wild animals such as commensal, savannah and forest-dwelling small mammals. The flea fauna of this zone is presented by X. cheopis and L. klossi. In the tropical forest surrounding villages four of the flea species were found: X. vexabilis, a specific parasite of the forest-dwelling rat Berylmys berdmorei, L. klossi found on six species of forest small mammals, Acropsylla girshami from Berylmys bowersii and Pariodontis subjugis from Hystrix brachyura. The agricultural zone is the most possible place of commensal and forest-dwelling small mammals contact, where the latter can get plague microbe.     

Out-of-Africa,human-mediated dispersal of the common cat flea,Ctenocephalides felis: The hitchhiker’s guide to world domination

The cat flea (Ctenocephalides felis) is the most common parasite of domestic cats and dogs worldwide. Due to the morphological ambiguity of C. felis and a lack of — particularly largescale — phylogenetic data, we do not know whether global C. felis populations are morphologically and genetically conserved, or whether human-mediated migration of domestic cats and dogs has resulted in homogenous global populations. To determine the ancestral origin of the species and to understand the level of global pervasion of the cat flea and related taxa, our study aimed to document the distribution and phylogenetic relationships of Ctenocephalides fleas found on cats and dogs worldwide. We investigated the potential drivers behind the establishment of regional cat flea populations using a global collection of fleas from cats and dogs across six continents. We morphologically and molecularly evaluated six out of the 14 known taxa comprising genus Ctenocephalides, including the four original C. felis subspecies (Ctenocephalides felis felis, Ctenocephalides felis strongylus, Ctenocephalides felis orientis and Ctenocephalides felis damarensis), the cosmopolitan species Ctenocephalides canis and the African species Ctenocephalides connatus. We confirm the ubiquity of the cat flea, representing 85% of all fleas collected (4357/5123). Using a multigene approach combining two mitochondrial (cox1 and cox2) and two nuclear (Histone H3 and EF-1α) gene markers, as well as a cox1 survey of 516 fleas across 56 countries, we demonstrate out-of-Africa origins for the genus Ctenocephalides and high levels of genetic diversity within C. felis. We define four bioclimatically limited C. felis clusters (Temperate, Tropical I, Tropical II and African) using maximum entropy modelling. This study defines the global distribution, African origin and phylogenetic relationships of global Ctenocephalides fleas, whilst resolving the taxonomy of the C. felis subspecies and related taxa. We show that humans have inadvertently precipitated the expansion of C. felis throughout the world, promoting diverse population structure and bioclimatic plasticity. By demonstrating the link between the global cat flea communities and their affinity for specific bioclimatic niches, we reveal the drivers behind the establishment and success of the cat flea as a global parasite.     

Control of flea populations in a simulated home environment model using lufenuron, imidacloprid or fipronil

Control strategies were evaluated over a 6-month period in a home simulation model comprising a series of similar carpeted pens, housing matched groups of six cats, in which the life-cycle of the flea Ctenocephalides felis felis Bouche (Siphonaptera: Pulicidae) had been established. Additional adult fleas were placed on the cats at intervals to mimic acquisition of extraneous fleas from outside the home. Treatment strategies included a single subcutaneous deposition of injectable lufenuron supported by initial treatments with a short-acting insecticidal spray, or monthly topical applications of imidacloprid or fipronil. An untreated control group indicated that conditions were suitable for flea replication and development. Controls had to be combed on 18 occasions to remove excessive flea burdens and two developed allergic reactions. Lufenuron cats were combed once and required two insecticidal treatments in the first month to achieve control. Even so, small flea burdens were constantly present thereafter. Imidacloprid and fipronil treatments appeared to give virtually complete control throughout. Single fleas were found on imidacloprid cats on two occasions, whereas none were recovered from fipronil cats at any time after the first treatment. Tracer cats were used to monitor re-infestation rates at the end of the trial period. Small numbers of host-seeking fleas were demonstrated in all treatment pens, indicating that total eradication had not been accomplished. It is concluded that the home environment simulation model incorporating tracer animals could provide a powerful tool for studying flea population dynamics under controlled conditions but improved techniques are needed for quantifying other off-host life-cycle stages.     

Comparison of nodulisporic acid analogs in a Lucilia sericata in vitro assay and a Ctenocephalides felis membrane feeding system

A medicinal chemistry program on the nodulisporic acid chemical class, guided by an artificial membrane flea-feeding assay, has recently identified permissive and nonpermissive regions of the pharmacophore for exploitation against fleas. This pathway was validated when several promising compounds from this program were administered orally to dogs at 15.0 mg/kg and found to have >90% flea activity for 2 wk. To determine if a surrogate insect assay would have provided the same guidance, a nodulisporic acid analog series was examined in both a Lucilia sericata larval assay and an artificial membrane flea-feeding assay using Ctenocephalides felis. Results from both insect assays were concordant in that even subtle chemical modification or substitution to the left-hand side of the nodulisporic acid pharmacophore resulted in substantial loss of insecticidal activity. Both assays were also in general agreement that the only modifications to the pharmacophore that did not result in loss of activity occurred to the C-8 side chain on the right-hand side of the molecule. Although there was good agreement between the 2 assays on the general regions of the pharmacophore, there was variance on individual compounds in the mono- and disubstituted amide series from the C-8 side chain. For example, the L. sericata assay showed several analogs from this subclass to possess similar activity to the parent acid, whereas the membrane assay indicated superior activity against fleas relative to the same parent. Consequently, although there was substantial general agreement between the assays, it was concluded that finer optimization of a lead compound should be done against the target parasite, even if it is ex vivo, as early as possible in a medicinal chemistry program.     

Detection of Rickettsia spp. and host and habitat associations of fleas (Siphonaptera) in eastern Taiwan

Rickettsia typhi and Rickettsia felis (Rickettsiales: Rickettsiaceae) are two rickettsiae principally transmitted by fleas, but the detection of either pathogen has rarely been attempted in Taiwan. Of 2048 small mammals trapped in eastern Taiwan, Apodemus agrarius Pallas (24.5%) and Mus caroli Bonhote (24.4%) (both: Rodentia: Muridae) were the most abundant, and M. caroli hosted the highest proportion of fleas (63.9% of 330 fleas). Two flea species were identified: Stivalius aporus Jordan and Rothschild (Siphonaptera: Stivaliidae), and Acropsylla episema Rothschild (Siphonaptera: Leptopsyllidae). Nested polymerase chain reaction targeting parts of the ompB and gltA genes showed six fleas to be positive for Rickettsia spp. (3.8% of 160 samples), which showed the greatest similarity to R. felis, Rickettsia japonica, Rickettsia conorii or Rickettsia sp. TwKM01. Rickettsia typhi was not detected in the fleas and Rickettsia co-infection did not occur. Both flea species were more abundant during months with lower temperatures and less rainfall, and flea abundance on M. caroli was not related to soil hardness, vegetative height, ground cover by litter or by understory layer, or the abundance of M. caroli. Our study reveals the potential circulation of R. felis and other rickettsiae in eastern Taiwan, necessitating further surveillance of rickettsial diseases in this region. This is especially important because many novel rickettsioses are emerging worldwide.     

Molecular detection of zoonotic bartonellae (B. henselae,B. elizabethae and B. rochalimae) in fleas collected from dogs in Israel

Fleas represent an acknowledged burden on dogs worldwide. The characterization of flea species infesting kennel dogs from two localities in Israel (Rehovot and Jerusalem) and their molecular screening for Bartonella species (Rhizobiales: Bartonellaceae) was investigated. A total of 355 fleas were collected from 107 dogs. The fleas were morphologically classified and molecularly screened targeting the Bartonella 16S–23S internal transcribed spacer (ITS). Of the 107 dogs examined, 80 (74.8%) were infested with Ctenocephalides canis (Siphonaptera: Pulicidae), 68 (63.6%) with Ctenocephalides felis, 15 (14.0%) with Pulex irritans (Siphonaptera: Pulicidae) and one (0.9%) with Xenopsylla cheopis (Siphonaptera: Pulicidae). Fleas were grouped into 166 pools (one to nine fleas per pool) according to species and host. Thirteen of the 166 flea pools (7.8%) were found to be positive for Bartonella DNA. Detected ITS sequences were 99–100% similar to those of four Bartonella species: Bartonella henselae (six pools); Bartonella elizabethae (five pools); Bartonella rochalimae (one pool), and Bartonella bovis (one pool). The present study indicates the occurrence of a variety of flea species in dogs in Israel; these flea species are, in turn, carriers of several zoonotic Bartonella species. Physicians, veterinarians and public health workers should be aware of the presence of these pathogens in dog fleas in Israel and preventive measures should be implemented.     

The absence of concordant population genetic structure in the black‐tailed prairie dog and the flea,Oropsylla hirsuta,with implications for the spread of Yersinia pestis

The black‐tailed prairie dog (Cynomys ludovicianus) is a keystone species on the mid‐ and short‐grass prairies of North America. The species has suffered extensive colony extirpations and isolation as a result of human activity including the introduction of an exotic pathogen, Yersinia pestis, the causative agent of sylvatic plague. The prairie dog flea, Oropsylla hirsuta, is the most common flea on our study colonies in north‐central Montana and it has been shown to carry Y. pestis. We used microsatellite markers to estimate the level of population genetic concordance between black‐tailed prairie dogs and O. hirsuta in order to determine the extent to which prairie dogs are responsible for dispersing this potential plague vector among prairie dog colonies. We sampled fleas and prairie dogs from six prairie dog colonies in two regions separated by about 46 km. These colonies were extirpated by a plague epizootic that began months after our sampling was completed in 2005. Prairie dogs showed significant isolation‐by‐distance and a tendency toward genetic structure on the regional scale that the fleas did not. Fleas exhibited higher estimated rates of gene flow among prairie dog colonies than the prairie dogs sampled from the same colonies. While the findings suggested black‐tailed prairie dogs may have contributed to flea dispersal, we attributed the lack of concordance between the population genetic structures of host and ectoparasite to additional flea dispersal that was mediated by mammals other than prairie dogs that were present in the prairie system.     

Grooming behaviors of black‐tailed prairie dogs are influenced by flea parasitism,conspecifics, and proximity to refuge

Grooming is a common animal behavior that aids in ectoparasite defense. Ectoparasites can stimulate grooming, and natural selection can also favor endogenous mechanisms that evoke periodic bouts of “programmed” grooming to dislodge or kill ectoparasites before they bite or feed. Moreover, grooming can function as a displacement or communication behavior. We compared the grooming behaviors of adult female black‐tailed prairie dogs (Cynomys ludovicianus) on colonies with or without flea control via pulicide dust. Roughly 91% of the prairie dogs sampled on the non‐dusted colony carried at least one flea, whereas we did not find fleas on two dusted colonies. During focal observations, prairie dogs on the non‐dusted colony groomed at higher frequencies and for longer durations than prairie dogs on the dusted colonies, lending support to the hypothesis that fleas stimulated grooming. However, the reduced amount of time spent grooming on the dusted colonies suggested that approximately 25% of grooming might be attributed to factors other than direct stimulation from ectoparasites. Non‐dusted colony prairie dogs rarely autogroomed when near each other. Dusted colony prairie dogs autogroomed for shorter durations when far from a burrow opening (refuge), suggesting a trade‐off between self‐grooming and antipredator defense. Allogrooming was detected only on the non‐dusted colony and was limited to adult females grooming young pups. Grooming appears to serve an antiparasitic function in C. ludovicianus. Antiparasitic grooming might aid in defense against fleas that transmit the plague bacterium Yersinia pestis. Plague was introduced to North America ca. 1900 and now has a strong influence on most prairie dog populations, suggesting a magnified effect of grooming on prairie dog fitness.     

Accumulation and persistence of flea larvicidal activity in the immediate environment of cats treated with imidacloprid

To investigate the persistence of flea larvicidal activity in the immediate environment of cats treated with imidacloprid, eggs of the cat flea Ctenocephalides felis felis Bouché (Siphonaptera: Pulicidae), from untreated donor cats, were incubated on samples of fleece blanket taken from the floor of cages used by treated or untreated cats for a total of 10 or 20 6-h periods over 2-4 weeks, respectively. Sufficient imidacloprid accumulated during these periods to reduce the emergence of adult fleas by 94.7-97.6% when the blankets were tested after 18 weeks' storage at room temperature. A typical laundry procedure (washing with detergent at 50 degrees C and low temperature tumble drying) removed this biological activity. Unwashed control blankets did not support the flea life-cycle as effectively as washed blankets or a sand substrate.