Colonization of a novel host by fleas: changes in egg production and egg size.

We studied the success of fleas, Synosternus cleopatrae and Xenopsylla ramesis, in switching to a novel host by establishing experimental lines maintained on different hosts for 18 generations. Fleas fed on principal (P-line) or novel hosts, either sympatric with (S-line) or allopatric to (A-line) a flea and its principal host, then we assessed their reproductive performance via the number and size of eggs. We compared reproductive performance between hosts within a line and between lines within a host asking: (a) whether fleas adapt to a novel host species after multiple generations; (b) if yes, whether the pattern of adaptation differs between novel host species sympatric with or allopatric to a flea and its principal host; and (c) adaptation to a novel host is accompanied with a loss of success in exploitation of an original host. Fleas from the S- and A-lines increased their egg production on a novel host (except X. ramesis from the S-line). S. cleopatrae from the S-line but not the A-line increased egg size on a novel host, whereas X. ramesis from the A-line but not the S-line produced larger eggs from a novel host. We found no indication of a loss of reproductive performance on the original host while adapting to a novel host. We conclude that fleas are able to switch rapidly to a new host with the pattern of a switch to either sympatric or an allopatric host depending on the identities of both flea and host species.

Feeding performance on a novel host: no adaptation over generations and differential patterns in two flea species.

To model the colonization of a novel host by fleas, Synosternus cleopatrae and Xenopsylla ramesis, we established experimental lines maintained for 15 generations on a principal or a novel host (either co-occurring with a flea or not). We compared the blood meal size and the energy expended for digestion by fleas from the 15th generation of each line on these hosts between hosts within a line and between lines within a host asking (a) whether fleas adapt to a novel host (increased blood consumption/decreased energy expended for digestion); (b) if yes, whether this adaptation leads to the loss of ability to exploit an original host, and (c) whether the success of adaptation to a novel host depends on its ecological co-occurrence with a flea. The blood consumption and digestion energetics of fleas fed on the principal host differed from those on other hosts. The effect of the principal host on feeding performance differed between fleas, with S. cleopatrae consuming less blood and expending more energy for digestion on the principal than on any other host, whereas the opposite was true for X. ramesis. No changes in feeding performance on a novel host over generations were found. We propose several explanations for the lack of adaptation to a novel host over time. We explain the poor performance of S. cleopatrae on its principal host via its immune response mounting pattern. We argue that the principal host of a parasite is not necessarily the host on which the parasite demonstrates the best performance.

Reproductive performance in generalist haematophagous ectoparasites: maternal environment, rearing conditions or both?

We tested whether and how the maternal environment (i.e. host species exploited by a mother), rearing conditions (i.e. host species exploited by her offspring) or both (i.e. matches and mismatches in host species exploited by a mother and her offspring) affect reproductive performance in the offspring. We experimentally manipulated maternal and rearing environments in two generalist fleas (Xenopsylla conformis and Xenopsylla ramesis) implementing a factorial cross-rearing design. Mothers exploited either the principal host (PH) or auxiliary hosts that were either closely (CAH) or distantly related (DAH) to the PH. After six generations of infesting a given host species, we cross-reared fleas within and between host species. These fleas reproduced and we measured their reproductive performance both quantitatively (i.e. egg number) and qualitatively (i.e. egg size, development time, body size of the next generation). We found that identity of the host a flea was reared on (=actual host) had the strongest effect on its performance. Individuals reared on the PH performed considerably better than those reared on either auxiliary host. Moreover, fleas reared on a CAH performed better than those reared on a DAH. Actual host identity also had a stronger effect on reproductive performance in X. ramesis than in X. conformis. Nevertheless, there was no difference in performance between match and mismatch maternal and actual host identities. We conclude that rearing environment has the strongest effect on fitness in generalist parasites. Moreover, phylogenetic distance between an auxiliary host and the PH determines the level of suitability of the former.

Morphological asymmetry and habitat quality: using fleas and their rodent hosts as a novel experimental system.

Morphological asymmetry is widely used to measure developmental instability and higher levels of asymmetry often correlate with decreased mating success, increased inbreeding, increased stress and decreased habitat quality. We studied asymmetry and relationships between asymmetry and host identity in two flea species, host generalist Xenopsylla ramesis and host specialist Parapulex chephrenis, and asked: (1) what the level of asymmetry was in their femurs and tibiae; (2) which type of asymmetry predominates; and (3) whether fleas that fed on host species distantly related to their principal host species produced offspring that exhibited greater asymmetry compared with offspring of fleas that fed on their principal host species. We found fluctuating asymmetry in femurs and tibiae of X. ramesis and in the tibiae of P. chephrenis as well as significantly left-handed directional asymmetry in the femurs of P. chephrenis Host species identity significantly impacted asymmetry in leg segments of P. chephrenis but not in those of X. ramesis Offspring asymmetry increased when mother fleas fed on a host that was distantly related to the principal host. Fleas parasitizing multiple host species might compensate for developmental instability when utilizing a novel host species; therefore, host-switching events in host-specific parasites could be constrained by the relatedness between a novel and a principal host species.

Effects of parasitism on host reproductive investment in a rodent-flea system: host litter size matters.

Parents may alter offspring phenotype depending on the type of environment they encounter. Parasitism is a common stressor; therefore, maternal reproductive investment could change in response to parasitic infection. However, few experiments have investigated the relationship between parasitism and maternal investment, whereas earlier field studies provided contradictory evidence. We investigated number, sex ratio, and growth of offspring in two rodent species, solitary altricial Meriones crassus and social precocial Acomys cahirinus, exposed to parasitism by fleas Xenopsylla ramesis and Parapulex chephrenis. No effect of treatment on litter size or sex ratio of a litter was found in either rodent species. Flea parasitism was found to affect pre-weaning body mass gain in M. crassus, but not in A. cahirinus pups. Furthermore, it appeared that female M. crassus invested resources into their offspring differently in dependence of litter size. In small litters (1-3 offspring), pups from infested females gained more body mass before weaning than pups from uninfested mothers. However, this trend was reversed in females with large litters indicating that parasitized females have a finite amount of resources with which to provision their young. Thus, M. crassus mothers parasitized by fleas seemed to receive some sort of external cues (e.g., stress caused by infestation) that prompted them to alter offspring provisioning, depending on species-specific possibilities and constraints. Therefore, parasites could be a mediator of environmentally induced maternal effects and offspring provisioning may have adaptive value against parasitism.

Flea fitness is reduced by high fractional concentrations of CO2 that simulate levels found in their hosts' burrows.

Nidicolous ectoparasites such as fleas and gamasid mites that feed on small and medium-sized mammals spend much of their time in their hosts' burrows, which provide an environment for living, and often feeding, to their pre-imaginal and/or adult stages. Thus, these ectoparasites should be adapted to environmental conditions in burrows, including high fractional concentrations of CO2 (F(CO2)). We examined how a high F(CO2) (0.04) affected survival and reproductive success of a hematophagous ectoparasite of burrowing rodents using fleas Xenopsylla ramesis and Sundevall's jirds Meriones crassus. In the first experiment, fleas fed on hosts housed in high-CO2 (F(CO2) =0.04) or atmospheric-CO2 (F(CO2) ≈0.0004) air, and were allowed to breed. In a second experiment, fleas were maintained in high CO2 or CO2-free air with no hosts to determine how CO2 levels affect survival and activity levels. We found that at high F(CO2) fleas laid fewer eggs, reducing reproductive success. In addition, at high F(CO2), activity levels and survival of fleas were reduced. Our results indicate that fleas do not perform well in the F(CO2) used in this experiment. Previous research indicated that the type and intensity of the effects of CO2 concentration on the fitness of an insect depend on the F(CO2) used, so we advise caution when generalizing inferences drawn to insects exposed to other F(CO2). If, however, F(CO2) found in natural mammal burrows brings about reduced fitness in fleas in general, then burrowing hosts may benefit from reduced parasite infestation if burrow air F(CO2) is high.

Fitness responses to co-infestation in fleas exploiting rodent hosts.

To understand mechanisms behind positive interspecific co-occurrences in flea infracommunities, we asked whether co-infestation results in an increase of flea fitness (quantity and/or quality of the offspring). We studied reproductive performance of Xenopsylla ramesis and Parapulex chephrenis when they exploited their characteristic host (Meriones crassus and Acomys cahirinus, respectively) either alone or together with another species. We used egg production, the number of new imagoes, pre-imaginal survival and egg size as fitness-related variables and predicted that fitness will be higher in fleas feeding in mixed- than in single-species groups. In both fleas, mean number of eggs produced per female flea did not depend on experimental treatment. No effect of single- vs mixed-species infestation on the mean number of new imagoes per female and the number of emerged imagoes per egg was found for X. ramesis, whereas both these numbers were higher in mixed- than in single-species groups for P. chephrenis. X. ramesis produced eggs of similar size independently of treatment, whereas eggs produced by P. chephrenis in mixed-species groups were significantly larger than eggs produced in single-species groups. We conclude that an increase in reproductive performance as a response to co-infestation may be one of the mechanisms behind aggregative structure of flea infracommunities. However, this response may vary among flea species.

Yersinia murine toxin is not required for early-phase transmission of Yersinia pestis by Oropsylla montana (Siphonaptera: Ceratophyllidae) or Xenopsylla cheopis (Siphonaptera: Pulicidae).

Plague, caused by Yersinia pestis, is characterized by quiescent periods punctuated by rapidly spreading epizootics. The classical 'blocked flea' paradigm, by which a blockage forms in the flea's proventriculus on average 1-2 weeks post-infection (p.i.), forces starving fleas to take multiple blood meals, thus increasing opportunities for transmission. Recently, the importance of early-phase transmission (EPT), which occurs prior to blockage formation, has been emphasized during epizootics. Whilst the physiological and molecular mechanisms of blocked flea transmission are well characterized, the pathogen-vector interactions have not been elucidated for EPT. Within the blocked flea model, Yersinia murine toxin (Ymt) has been shown to be important for facilitating colonization of the midgut within the flea. One proposed mechanism of EPT is the regurgitation of infectious material from the flea midgut during feeding. Such a mechanism would require bacteria to colonize and survive for at least brief periods in the midgut, a process that is mediated by Ymt. Two key bridging vectors of Y. pestis to humans, Oropsylla montana (Siphonaptera: Ceratophyllidae) or Xenopsylla cheopis (Siphonaptera: Pulicidae), were used in our study to test this hypothesis. Fleas were infected with a mutant strain of Y. pestis containing a non-functional ymt that was shown previously to be incapable of colonizing the midgut and were then allowed to feed on SKH-1 mice 3 days p.i. Our results show that Ymt was not required for EPT by either flea species.

Ectoparasite performance when feeding on reproducing mammalian females: an unexpected decrease when on pregnant hosts.

Reproduction is an energy-demanding activity in mammalian females, with increased energy requirements during pregnancy and, especially, during lactation. To better understand the interactions between parasitism and host reproduction, we investigated feeding and reproductive performance of fleas (Xenopsylla ramesis) parasitizing non-reproducing, pregnant or lactating gerbilline rodents (Meriones crassus). Based on energetic considerations, we predicted that feeding and reproductive performance of fleas would be lowest on non-breeding females, moderate on pregnant females and highest on lactating females. We estimated feeding performance of the fleas via absolute and mass-specific bloodmeal size and reproductive performance via egg production and latency to peak oviposition. Host reproductive status had no effect on either absolute or mass-specific bloodmeal size or the day of peak oviposition, but significantly affected the daily number of eggs produced by a female flea. Surprisingly, and contrary to our predictions, egg production of fleas fed on pregnant rodents was significantly lower than that of fleas on non-reproducing and lactating rodents, while no difference in egg production between fleas feeding on non-reproducing and lactating hosts was found. Our results suggest that differences in parasite reproduction when feeding on hosts of different reproductive status are not associated with the different energy requirements of the hosts at non-breeding, pregnancy and lactation but rather with variation in hormonal and/or immune status during these periods.

A trade-off between quantity and quality of offspring in haematophagous ectoparasites: the effect of the level of specialization.

Theory predicts an adaptive trade-off between quantity and quality of offspring if mothers can reliably predict the offspring environment. We studied egg production and quality of offspring in two flea species (host-specialist Parapulex chephrenis and host-generalist Xenopsylla ramesis) exploiting eight rodent species. We evaluated quality of new imagoes via their developmental time, size (length of a femur as a proxy) and resistance to starvation without a blood meal. We predicted that the offspring quality would increase with (i) a decrease in the number of eggs produced by mothers and (ii) an increase in phylogenetic distance between maternal host and principal host of a flea. We also predicted that negative relationships between offspring quality and either maternal egg production effort or phylogenetic distance between maternal host and the principal host or both would be manifested stronger in host-opportunistic than in host-specific fleas. The highest number of eggs produced per female flea was accompanied by the longest duration of development and the smallest offspring in X. ramesis, while P. chephrenis that hatched from larger clutches survived for less time under starvation. Although there was no significant effect of host species on any dependent variable, association between offspring quality and phylogenetic distance of the maternal host from the principal host of a flea was found in X. ramesis (but not P. chephrenis) with new imagoes being larger if their maternal hosts were phylogenetically distant from the principal host. Our results demonstrated stronger trade-off between quantity and quality of offspring in a generalist than in a specialist flea, supporting the association between life-history plasticity and generalist feeding strategy.

Effects of Bartonella spp. on flea feeding and reproductive performance.

Numerous pathogens are transmitted from one host to another by hematophagous insect vectors. The interactions between a vector-borne organism and its vector vary in many ways, most of which are yet to be explored and identified. These interactions may play a role in the dynamics of the infection cycle. One way to evaluate these interactions is by studying the effects of the tested organism on the vector. In this study, we tested the effects of infection with Bartonella species on fitness-related variables of fleas by using Bartonella sp. strain OE 1-1, Xenopsylla ramesis fleas, and Meriones crassus jirds as a model system. Feeding parameters, including blood meal size and metabolic rate during digestion, as well as reproductive parameters, including fecundity, fertility, and life span, were compared between fleas experimentally infected with Bartonella and uninfected fleas. In addition, the developmental time, sex ratio, and body size of F1 offspring fleas were compared between the two groups. Most tested parameters did not differ between infected and uninfected fleas. However, F1 males produced by Bartonella-positive females were significantly smaller than F1 males produced by Bartonella-negative female fleas. The findings in this study suggest that bartonellae are well adapted to their flea vectors, and by minimally affecting their fitness they have evolved to better spread themselves in the natural environment.

[The systemic effects of imidacloprid and fipronil on Xenopsylla cheopis fleas (Siphonaptera) and blood-sucking Ornithonyssus bacoti gamasid mites (Gamasina: Macronyssidae)].

The systemic effects of imidacloprid and fipronil on Xenopsylla cheopis fleas and blood-sucking Ornithonyssus bacoti gamasid mites were studied under laboratory conditions. Albino rats were forcibly administered insectoacaricides per os in doses of 1, 10, 30 mg/kg or fed with dietary bait (the doses of fipronil and imidacloprid were 10-33 and 18-70 mg/kg/day, respectively) for 3 or more days. Fipronil had a pronounced systemic effect on both X. cheopis and O. baconi. Imidacloprid was markedly active against the fleas and, on the contrary, inactive against the mites.

Use it or lose it: reproductive implications of ecological specialization in a haematophagous ectoparasite.

Using experimentally induced disruptive selection, we tested two hypotheses regarding the evolution of specialization in parasites. The 'trade-off' hypothesis suggests that adaptation to a specific host may come at the expense of a reduced performance when exploiting another host. The alternative 'relaxed selection' hypothesis suggests that the ability to exploit a given host would deteriorate when becoming obsolete. Three replicate populations of a flea Xenopsylla ramesis were maintained on each of two rodent hosts, Meriones crassus and Dipodillus dasyurus, for nine generations. Fleas maintained on a specific host species for a few generations substantially decreased their reproductive performance when transferred to an alternative host species, whereas they generally did not increase their performance on their maintenance host. The results support the 'relaxed selection' hypothesis of the evolution of ecological specialization in haematophagous ectoparasites, while suggesting that trade-offs are unlikely drivers of specialization. Further work is needed to study the extent by which the observed specializations are based on epigenetic or genetic modifications.

Effects of host diet and thermal state on feeding performance of the flea Xenopsylla ramesis.

We examined feeding performance of the flea Xenopsylla ramesis on three different hosts: its natural, granivorous, rodent host, Sundevall's jird (Meriones crassus); the frugivorous Egyptian fruit bat (Rousettus aegyptiacus); and an insectivorous bat, Kuhl's pipistrelle (Pipistrellus kuhlii). Because these fleas are not known to occur on bats, we hypothesized that the fleas' feeding performance (i.e. feeding and digestion rates) would be higher when feeding on their natural host than on either of the bats that they do not naturally parasitize. We found that mass-specific blood-meal size of both male and female fleas was significantly lower when feeding on Kuhl's pipistrelles than on the other two species, but was not different in female fleas feeding on fruit bats or on jirds at all stages of digestion. However, more male fleas achieved higher levels of engorgement if they fed on Sundevall's jirds than if they fed on Egyptian fruit bats. The fleas digested blood of fruit bats and jirds significantly faster than blood of Kuhl's pipistrelle. In addition, after a single blood meal, the survival time of fleas fed on normothermic Kuhl's pipistrelles was significantly shorter than that of fleas fed on Sundevall's jirds and even lower when male fleas fed on Egyptian fruit bats. Thus, our prediction was partially supported: normothermic Kuhl's pipistrelles were inferior hosts for fleas compared with Sandevall's jirds and Egyptian fruit bats. Interestingly, the proportion of engorged fleas that fed on torpid Kuhl's pipistrelles was significantly higher than the proportion of the fleas that fed on normothermic individuals, indicating that becoming torpid might be a liability, rather than an effective defense against parasites.

Investigation of Bartonella acquisition and transmission in Xenopsylla ramesis fleas (Siphonaptera: Pulicidae).

Bartonella are emerging and re-emerging pathogens affecting humans and a wide variety of animals including rodents. Horizontal transmission of Bartonella species by different hematophagous vectors is well acknowledged but vertical transmission (from mother to offspring) is questionable and was never explored in fleas. The aim of this study was to investigate whether the rodent flea, Xenopsylla ramesis, can acquire native Bartonella from wild rodents and transmit it transovarially. For this aim, Bartonella-free laboratory-reared X. ramesis fleas were placed on six naturally Bartonella-infected rodents and six species-matched Bartonella-negative rodents (three Meriones crassus jirds, two Gerbillus nanus gerbils and one Gerbillus dasyurus gerbil) for 7 days, 12-14h per day. The fleas that were placed on the Bartonella-positive rodents acquired four different Bartonella genotypes. Eggs and larvae laid and developed, respectively, by fleas from both rodent groups were collected daily for 7 days and molecularly screened for Bartonella. All eggs and larvae from both groups were found to be negative for Bartonella DNA. Interestingly, two of five gut voids regurgitated by Bartonella-positive fleas contained Bartonella DNA. The naturally infected rodents remained persistently infected with Bartonella for at least 89 days suggesting their capability to serve as competent reservoirs for Bartonella species. The findings in this study indicate that X. ramesis fleas can acquire several Bartonella strains from wild rodents but cannot transmit Bartonella transovarially.

Effects of temperature on early-phase transmission of Yersina pestis by the flea, Xenopsylla cheopis.

Sharp declines in human and animal cases of plague, caused by the bacterium Yersinia pestis (Yersin), have been observed when outbreaks coincide with hot weather. Failure of biofilm production, or blockage, to occur in the flea, as temperatures reach 30 degrees C has been suggested as an explanation for these declines. Recent work demonstrating efficient flea transmission during the first few days after fleas have taken an infectious blood meal, in the absence of blockage (e.g., early-phase transmission), however, has called this hypothesis into question. To explore the potential effects of temperature on early-phase transmission, we infected colony-reared Xenopsylla cheopis (Rothchild) fleas with a wild-type strain of plague bacteria using an artificial feeding system, and held groups of fleas at 10, 23, 27, and 30 degrees C. Naive Swiss Webster mice were exposed to fleas from each of these temperatures on days 1-4 postinfection, and monitored for signs of infection for 21 d. Temperature did not significantly influence the rates of transmission observed for fleas held at 23, 27, and 30 degrees C. Estimated per flea transmission efficiencies for these higher temperatures ranged from 2.32 to 4.96% (95% confidence interval [CI]: 0.96-8.74). In contrast, no transmission was observed in mice challenged by fleas held at 10 degrees C (per flea transmission efficiency estimates, 0-1.68%). These results suggest that declines in human and animal cases during hot weather are not related to changes in the abilities of X. cheopis fleas to transmit Y. pestis infections during the early-phase period. By contrast, transmission may be delayed or inhibited at low temperatures, indicating that epizootic spread of Y. pestis by X. cheopis via early-phase transmission is unlikely during colder periods of the year.

Host gender and offspring quality in a flea parasitic on a rodent.

The quality of offspring produced by parent fleas (Xenopsylla ramesis) fed on either male or female rodent hosts (Meriones crassus) was studied. The emergence success, duration of development, resistance to starvation upon emergence and body size of the flea offspring were measured. It was predicted that offspring of fleas produced by parents that fed on male hosts (i) will survive better as pre-imago, (ii) will develop faster, (iii) will live longer under starvation after emergence and (iv) will be larger than offspring of fleas fed on female hosts. The emergence success of pre-imaginal fleas was relatively high, ranging from 46.9% to 100.0% and averaging 78.4±3.0%, and was not affected by host gender. The duration of development of pre-imaginal fleas depended on the gender of the host of parents and differed between male and female offspring, with female fleas developing faster. Furthermore, male fleas developed faster if their parents fed on female rather than on male hosts, whereas no difference in the duration of development between host genders was found in female fleas. The time to death under starvation did not depend on the gender of either the flea or the host. A newly emerged flea, on average, lived 31.9±1.0 days without access to food. The relationship between host gender and body size of male flea offspring was the only effect that supported the predictions. An increase in body size in male fleas could increase their mating success and, ultimately, their fitness.

An Evaluation of the Flea Index as a Predictor of Plague Epizootics in the West Nile Region of Uganda.

Plague is a low incidence flea-borne zoonosis that is often fatal if treatment is delayed or inadequate. Outbreaks occur sporadically and human cases are often preceded by epizootics among rodents. Early recognition of epizootics coupled with appropriate prevention measures should reduce plague morbidity and mortality. For nearly a century, the flea index (a measure of fleas per host) has been used as a measure of risk for epizootic spread and human plague case occurrence, yet the practicality and effectiveness of its use in surveillance programs has not been evaluated rigorously. We sought to determine whether long-term monitoring of the Xenopsylla flea index on hut-dwelling rats in sentinel villages in the plague-endemic West Nile region of Uganda accurately predicted plague occurrence in the surrounding parish. Based on observations spanning ~6 yr, we showed that on average, the Xenopsylla flea index increased prior to the start of the annual plague season and tended to be higher in years when plague activity was reported in humans or rodents compared with years when it was not. However, this labor-intensive effort had limited spatial coverage and was a poor predictor of plague activity within sentinel parishes.

Presence of the Oriental Rat Flea (Siphonaptera: Pulicidae) Infesting an Endemic Mammal and Confirmed Plague Circulation in a Forest Area of Madagascar.

The Oriental rat flea, Xenopsylla cheopis (Rothschild 1903), is a cosmopolitan flea usually found infesting domestic rats. This flea is a well-known major human plague vector in Madagascar. As part of field sampling, fleas and small mammals were collected in the village of South Andranofeno and the natural reserve of Sohisika, two sites of the district of Ankazobe, located in the Central Highlands of Madagascar. Rats inside houses and forest small mammals were trapped using Besancon Technical Services and pitfall traps, respectively. Their fleas were collected and preserved for laboratory works. Collected fleas from the village and forest belonged to five species, which were X. cheopis, Synopsyllus fonquerniei (Wagner and Roubaud 1932) (Siphonaptera: Pulicidae), Echidnophaga gallinacea (Westwood 1875) (Siphonaptera: Pulicidae), Ctenocephalides felisstrongylus (Jordan 1925) (Siphonaptera: Pulicidae), Pulex irritans (Linnaeus 1758) (Siphonaptera: Pulicidae). After sampling in the forest zone, one specimen of X. cheopis was unexpectedly collected while infesting an endemic tenrec Setifer setosus (Schreber 1777) (Afrosoricida: Tenrecidae). Polymerase chain reaction (PCR) diagnosis on all collected fleas allowed detecting plague bacterium Yersinia pestis (Lehmann and Neumann 1896) (Enterobacterales: Yersiniaceae) on nine specimens of the endemic flea S. fonquerniei collected inside forest. The presence of the oriental rat flea in forest highlights the connection between human and wild environments due to animal movements and the fact that the rat flea can infest various hosts. As only one specimen of X. cheopis was collected on S. setosus, we hypothesize that flea was carried from the village to forest. Yersinia pestis infection of forest fleas outlines plague circulation in this sylvatic area.

Genetic differentiation of the oriental rat flea, Xenopsylla cheopis, from two sympatric host species.

BACKGROUND: The oriental rat flea (Xenopsylla cheopis), which infests several mammals, primarily rats (Rattus spp.), is the most notorious vector of human plague. In this study, we measured the genetic differentiation among populations of fleas from the Asian house rat (Rattus tanezumi) and the brown rat (R. norvegicus) using microsatellite markers in order to investigate the extent of host-switching in this parasite. RESULTS: We developed 11 polymorphic microsatellite loci for our study, nine of which showed high potential for inbreeding. AMOVA showed that the majority (84.07%, P < 0.001) of the variation was derived from within populations, followed by variation among groups (14.96%, P < 0.001); in contrast, variation within groups of populations was nearly absent (0.97%, P > 0.05). Analyses of the pairwise fixation index revealed that most of the ten allopatric population pairs but none of the five sympatric population pairs were significantly differentiated. Moreover, based on genetic structure clustering analysis, there was obvious differentiation between allopatric populations but not between sympatric population pairs. CONCLUSIONS: These results indicate the presence of frequent migrations of the oriental rat flea between the sympatric Asian house rat and brown rat, causing a high rate of gene flow and limited genetic differentiation. We suggest that there is no clear boundary limiting the migration of oriental rat fleas between the two hosts, and thus both rat species should be monitored equally for the purposes of plague prevention and control.