Search

Few studies have been able to provide experimental evidence of the ability of fleas to maintain rodent-associated Bartonella infections and excrete these bacteria. These data are important for understanding the transmission cycles and prevalence of these bacteria in hosts and vectors. We used an artificial feeding approach to expose groups of the oriental rat flea (Xenopsylla cheopis Rothschild; Siphonaptera, Pulicidae) to rat blood inoculated with varying concentrations of Bartonella elizabethae Daly (Bartonellaceae: Rhizobiales). Flea populations were maintained by membrane feeding on pathogen-free bloodmeals for up to 13 d post infection. Individual fleas and pools of flea feces were tested for the presence of Bartonella DNA using molecular methods (quantitative and conventional polymerase chain reaction [PCR]). The threshold number of Bartonellae required in the infectious bloodmeal for fleas to be detected as positive was 106 colony-forming units per milliliter (CFU/ml). Individual fleas were capable of harboring infections for at least 13 d post infection and continuously excreted Bartonella DNA in their feces over the same period. This experiment demonstrated that X. cheopis are capable of acquiring and excreting B. elizabethae over several days. These results will guide future work to model and understand the role of X. cheopis in the natural transmission cycle of rodent-borne Bartonella species. Future experiments using this artificial feeding approach will be useful for examining the horizontal transmission of B. elizabethae or other rodent-associated Bartonella species to naïve hosts and for determining the viability of excreted bacteria.

Rodent fleas from northwestern Chihuahua, Mexico, were analyzed for the presence of Bartonella and Yersinia pestis. In total, 760 fleas belonging to 10 species were tested with multiplex polymerase chain reaction analysis targeting the gltA (338-bp) and pla genes (478-bp) of Bartonella and Y. pestis, respectively. Although none was positive for Y. pestis, 307 fleas were infected with Bartonella spp., resulting in an overall prevalence of 40.4%. A logistic regression analysis indicated that the presence of Bartonella is more likely to occur in some flea species. From a subset of Bartonella-positive fleas, phylogenetic analyses of gltA gene sequences revealed 13 genetic variants clustering in five phylogroups (I–V), two of which were matched with known pathogenic Bartonella species (Bartonella vinsonii subsp. arupensis and Bartonella washoensis) and two that were not related with any previously described species or subspecies of Bartonella. Variants in phylogroup V, which were mainly obtained from Meringis spp. fleas, were identical to those reported recently in their specific rodent hosts (Dipodomys spp.) in the same region, suggesting that kangaroo rats and their fleas harbor other Bartonella species not reported previously. Considering the Bartonella prevalence and the flea genotypes associated with known pathogenic Bartonella species, we suggest that analysis of rodent and flea communities in the region should continue for their potential implications for human health. Given that nearby locations in the United States have reported Y. pestis in wild animals and their fleas, we suggest conducting larger-scale studies to increase our knowledge of this bacterium.

Early-phase transmission (EPT) of Yersinia pestis by unblocked fleas is a well-documented, replicable phenomenon with poorly defined mechanisms. We review evidence demonstrating EPT and current knowledge on its biological and biomechanical processes. We discuss the importance of EPT in the epizootic spread of Y. pestis and its role in the maintenance of plague bacteria in nature. We further address the role of EPT in the epidemiology of plague., (Published by Oxford University Press on behalf of Entomological Society of America 2015. This work is written by US Government employees and is in the public domain in the US.)

Plague is a primarily flea-borne rodent-associated zoonosis that is often fatal in humans. Our study focused on the plague-endemic West Nile region of Uganda where affordable means for the prevention of human plague are currently lacking. Traditional hut construction and food storage practices hinder rodent exclusion efforts, and emphasize the need for an inexpensive but effective host-targeted approach for controlling fleas within the domestic environment. Here we demonstrate the ability of an insecticide delivery tube that is made from inexpensive locally available materials to reduce fleas on domestic rodents. Unbaited tubes were treated with either an insecticide alone (fipronil) or in conjunction with an insect growth regulator [(S)-methoprene], and placed along natural rodent runways within participant huts. Performance was similar for both treatments throughout the course of the study, and showed significant reductions in the proportion of infested rodents relative to controls for at least 100 d posttreatment., (© 2014 Entomological Society of America.)

Plague, a primarily flea-borne disease caused by Yersinia pestis, is characterized by rapidly spreading epizootics separated by periods of quiescence. Little is known about how and where Y. pestis persists between epizootics. It is commonly proposed, however, that Y pestis is maintained during interepizootic periods in enzootic cycles involving flea vectors and relatively resistant host populations. According to this model, while susceptible individuals serve as infectious sources for feeding fleas and subsequently die of infection, resistant hosts survive infection, develop antibodies to the plague bacterium, and continue to provide bloodmeals to infected fleas. For Y. pestis to persist under this scenario, fleas must remain infected after feeding on hosts carrying antibodies to Y. pestis. Studies of other vector-borne pathogens suggest that host immunity may negatively impact pathogen survival in the vector. Here, we report infection rates and bacterial loads for fleas (both Xenopsylla cheopis (Rothschild) and Oropsylla montana (Baker)) that consumed an infectious bloodmeal and subsequently fed on an immunized or age-matched naive mouse. We demonstrate that neither the proportion of infected fleas nor the bacterial loads in infected fleas were significantly lower within 3 d of feeding on immunized versus naive mice. Our findings thus provide support for one assumption underlying the enzootic host model of interepizootic maintenance of Y. pestis.

Programs that aim to control vector-borne zoonotic diseases require information on zoonotic hosts and on the feeding behavior of bridging vectors that are capable of transmitting pathogens from those hosts to humans. Here we describe an assay developed to identify bloodmeals in field-collected cat fleas (Ctenocephalides felis Bouché) to assess this species' potential role as a Yersinia pestis bridging vector in a plague-endemic region of Uganda. Our assay uses a single primer set and SYBR Green I-based real-time polymerase chain reaction to amplify a segment of the 12S mitochondrial ribosomal RNA gene for identification by sequencing. The assay capitalizes on the sensitivity of real-time polymerase chain reaction and the specificity of sequencing and can be used to differentiate vertebrate bloodmeals to the genus or species level without a priori knowledge of the host community. Because real-time assays that detect vertebrate DNA are highly sensitive to human DNA contamination, we analyzed detection in artificially fed and unfed fleas to establish a Ct cutoff that optimized specificity without completely sacrificing sensitivity. Using the established cutoff, our assay detected human, rat, and goat DNA in artificially fed C. felis up to 72 h postfeeding.

Over the past two decades, the majority of human plague cases have been reported from areas in Africa, including Uganda. In an effort to develop affordable plague control methods within an integrated vector control framework, we evaluated the efficacy of indoor residual spraying (IRS) techniques commonly used for mosquito control for controlling fleas on hut-dwelling commensal rodents in a plague-endemic region of Uganda. We evaluated both the standard IRS spraying (walls and ceiling) and a modified IRS technique that included insecticide application on not only on walls and ceiling but also a portion of the floor of each treated hut. Our study demonstrated that both the standard and modified IRS applications were effective at significantly reducing the flea burden and flea infestation of commensal rodents for up to 100 d after application, suggesting that IRS could potentially provide simultaneous control of mosquito and fleaborne diseases.

We used high-throughput DNA sequencing to explore bacterial communities of three species of Oropyslla fleas [Oropsylla hirsuta (Baker), Oropsylla montana (Baker), and Oropsylla tuberculata cynomuris (Jellison)] and detected seven bacterial lineages related to known insect symbionts. No significant co-occurrence patterns were detected among bacterial lineages, but relative abundance data suggest that the two most common lineages (Bartonella and Rickettsiales) interact negatively. Furthermore, presence of these two lineages significantly reduced bacterial diversity within fleas.

Quantifying the abundance of host-seeking fleas is critical for assessing risk of human exposure to flea-borne disease agents, including Yersinia pestis, the etiological agent of plague. Yet, reliable measures of the efficacy of existing host-seeking flea collection methods are lacking. In this study, we compare the efficacy of passive and active methods for the collection of host-seeking fleas in both the laboratory and human habitations in a plague-endemic region of northwest Uganda. In the laboratory, lighted "Kilonzo" flea traps modified with either blinking lights, the creation of shadows or the generation of carbon dioxide were less efficient at collecting Xenopsylla cheopis Rothchild and Ctenocephalides felis Bouché fleas than an active collection method using white cotton socks or cotton flannel. Passive collection using Kilonzo light traps in the laboratory collected significantly more X. cheopis than C. felis and active collection, using white socks and flannel, collected significantly more C. felis than X. cheopis. In field studies conducted in Uganda, Kilonzo traps using a flashlight were similar in their collection efficacy to Kilonzo traps using kerosene lamps. However, in contrast to laboratory studies, Kilonzo flea traps using flashlights collected a greater number of fleas than swabbing. Within human habitations in Uganda, Kilonzo traps were especially useful for collecting C. felis, the dominant species found in human habitations in this area.

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.

In recent decades, the majority of human plague cases (caused by Yersinia pestis) have been reported from Africa. In an effort to reduce the risk of the disease in this area, we evaluated the efficacy of a host-targeted rodent bait containing the insecticide imidacloprid for controlling fleas on house-dwelling commensal rodents in a plague-endemic region of northwestern Uganda. Results demonstrated that the use of a palatable, rodent-targeted, wax-based bait cube was effective at reducing the prevalence of fleas on commensal rodents and flea burdens on these animals at day 7 postbait exposure, but lacked significant residual activity, allowing flea populations to rebound in the absence of additional bait applications. Our results indicate the use of a palatable host-targeted bait block containing imidacloprid was an effective technique for quickly reducing flea numbers on rodents in northwest Uganda and, thus, could be useful for lowering the potential risk of human flea bite exposures during plague outbreaks if applied continuously during the period of risk.

The etiological agent of plague, Yersinia pestis, is most commonly transmitted by the bite of infectious fleas. To date, at least 28 flea species occurring in North America have been experimentally confirmed as vectors of Y. pestis. Transmission efficiency differs among species and also between different studies of a single species. These differences may, however, in large part reflect nonstandardized experimental conditions used during the first half of the 20th century when such studies were conducted in response to the rapid spread of Y. pestis across the western United States after its introduction at the beginning of this century. The majority of these early transmission studies focused on the blocked flea mechanism of transmission, which typically does not occur until > 2-3 wk after the flea becomes infected. Recent studies have challenged the paradigm that Y. pestis is usually spread by blocked fleas by demonstrating that numerous flea species, including the oriental rat flea Xenopsylla cheopis, which was the focus of the early classical studies on blocked flea transmission, are capable of"early-phase" transmission during the first few days after becoming infected and before a complete blockage can form. The aims of this review are to 1) summarize Y. pestis vector competency and efficiency studies for fleas occurring in North America, 2) discuss the implications of the results of these studies for our understanding of the dynamics of plague epizootics, 3) demonstrate why older transmission studies need to be repeated using a standardized experimental system, and 4) outline future directions for studies of fleas as vectors of Y. pestis.

The role of deer mice and other species of Peromyscus as enzootic reservoirs for plague remains controversial. In this study, we evaluated early-phase vector efficiency of Aetheca wagneri Baker, a common flea species infesting deer mice, to determine the likelihood that Y. pestis could be spread mouse to mouse by this species. We showed that A. wagneri could transmit plague bacteria to laboratory mice as early as 3 d postinfection (p.i.), but transmission efficiency was quite low (1.03%; 95% CI: 0.19-3.34%) 1-4 d p.i. compared with that for the established plague vector Oropsylla montana Baker (10.63%; 95% CI: 4.18-25.91). Using this early-phase transmission efficiency estimate, we determined through parameterization of a simple predictive model that at least 68 A. wagneri per deer mouse would be required to support levels of transmission adequate for enzootic maintenance. Because deer mice typically harbor fewer than three A. wagneri per host, our data do not support the notion of an independent deer mouse--A. wagneri transmission cycle.

Yersinia pestis, the etiological agent of plague, is transmitted by multiple flea species. Previous studies have reported wide variability in transmission efficiency among competent vectors. However, it is unclear to what extent such variation is explained by methodological differences among studies. To optimize an artificial feeding system where fleas are infected with controlled numbers of Y. pestis under standardized laboratory conditions that could be used to systematically compare vector efficiency, we sought to test the effect of host bloodmeal source on (1) the flea's ability to remain infected with Y. pestis and (2) bacterial loads in fleas. Here, we demonstrate that both prevalence of infection with a virulent strain of Y. pestis (CO96-3188) and bacterial loads in rock squirrel fleas (Oropsylla montana) are affected by host-associated blood factors. The generality of this observation was confirmed by repeating the study using the rat flea (Xenopsylla cheopis) and a commonly used avirulent laboratory strain of Y. pestis (A1122). Implications of the results for rate of spread of Y. pestis in naturally infected host populations are discussed.

Plague, a flea-borne zoonotic disease, is characterized by rapidly spreading epizootics. Rate of infectious spread is thought to be related to daily biting rate of the vector, the extrinsic incubation period, vector efficiency, and the duration of infectivity. A recent study of Oropsylla montana (Baker) (Siphonaptera: Ceratophyllidae), the primary vector of Yersinia pestis (Yersin) to humans in North America, revealed that this flea feeds readily on a daily basis, has a very short extrinsic incubation period, and efficiently transmits plague bacteria for at least 4 d postinfection (p.i.). Earlier studies based on fleas receiving a single infectious bloodmeal showed that transmission efficiency wanes after 4 d p.i. In our study, we simulate a naturally occurring scenario in which fleas are exposed repeatedly to septicemic hosts, and we evaluate vector efficiency of O. montana 6-9 d after the initial infectious bloodmeal for 1) fleas given a "booster" infectious bloodmeal 5 d after initial exposure and 2) fleas that received an uninfected maintenance bloodmeal 5 d p.i. Transmission of Y. pestis was not observed beyond 7 d after initial exposure in the fleas that received a single infectious bloodmeal, whereas fleas given a booster infectious bloodmeal could transmit throughout the 9-d duration of the study. The proportion of flea pools transmitting Y. pestis was significantly higher for fleas receiving multiple, rather than single infectious bloodmeals. Surprisingly, transmission success was not directly related to bacterial loads in fleas. Our data indicated that the duration of time over which O. montana reliably transmitted plague bacteria was longer than previously thought, and this may help to explain rapid rates of epizootic spread.

For almost a century, the oriental rat flea, Xenopsylla cheopis (Rothschild) (Siphonaptera: Pulicidae), was thought to be the most efficient vector of the plague bacterium Yersinia pestis (Yersin). Approximately 2 wk after consuming an infectious bloodmeal, a blockage often forms in the flea's proventriculus, which forces the flea to increase its biting frequency and consequently increases the likelihood of transmission. However, if fleas remain blocked and continue to feed, they usually die within 5 d of blocking, resulting in a short infectious window. Despite observations of X. cheopis transmitting Y. pestis shortly after pathogen acquisition, early-phase transmission (e.g., transmission 1-4 d postinfection [ p.i.]) by unblocked fleas was viewed as anomalous and thought to occur only by mass action. We used an artificial feeding system to infect colony-reared X. cheopis with a fully virulent strain of Y. pestis, and we evaluated transmission efficiency 1- 4 d p.i. We demonstrate 1) that a single infected and unblocked X. cheopis can infect a susceptible host as early as 1 d p.i., 2) the number of fleas per host required for unblocked fleas to drive a plague epizootic by early-phase transmission is within the flea infestation range observed in nature, and 3) early-phase transmission by unblocked fleas in the current study was at least as efficient as transmission by blocked fleas in a previously published study using the same colony of fleas and same bacterial strain. Furthermore, transmission efficiency seemed to remain constant until block formation, resulting in an infectious period considerably longer than previously thought.

Plague is a rare but highly virulent flea-borne zoonotic disease caused by the Gram-negative bacterium Yersinia pestis Yersin. Identifying areas at high risk of human exposure to the etiological agent of plague could provide a useful tool for targeting limited public health resources and reduce the likelihood of misdiagnosis by raising awareness of the disease. We created logistic regression models to identify landscape features associated with areas where humans have acquired plague from 1957 to 2004 in the four-corners region of the United States (Arizona, Colorado, New Mexico, and Utah), and we extrapolated those models within a geographical information system to predict where plague cases are likely to occur within the southwestern United States disease focus. The probability of an area being classified as high-risk plague habitat increased with elevation up to approximately 2300 m and declined as elevation increased thereafter, and declined with distance from key habitat types (e.g., southern Rocky Mountain piñon--juniper [Pinus edulis Engelm. and Juniperus spp.], Colorado plateau piñon--juniper woodland, Rocky Mountain ponderosa pine (Pinus ponderosa P.& C. Lawson var. scopulorum), and southern Rocky Mountain juniper woodland and savanna). The overall accuracy of the model was >82%. Our most conservative model predicted that 14.4% of the four-corners region represented a high risk of peridomestic exposure to Y. pestis.

The flea and rodent samples studied in this project were collected from field study sites in New Mexico from winter 1998 to spring 2001. During this period, 155 small rodents (14 different species) were live-trapped and combed for the presence of fleas. A total of 253 fleas were collected, comprising 21 species. Two of the 253 fleas collected were polymerase chain reaction (PCR) positive for the Rickettsia 17-kDa protein gene. These two fleas were both Anomiopsyllus nudata Baker, each collected from an individual Neotoma albigula Hartley, on two occasions. Individual fleas positive for the Rickettsia 17-kDa protein gene were then tested with primers targeting the rickettsial genes for citrate synthase (gltA) and two major outer membrane proteins (ompA and ompB). The nucleotide sequences of the PCR products of these two fleas were identical to each other and were 100% (394/394), 100% (1150/1150), 99.8% (469/470), and 99.3% (818/824) similar to the corresponding sequences of the 17-kDa, gltA, ompA, and ompB genes of Rickettsia felis, respectively. Flea homogenates of individual PCR-positive fleas were inoculated into shell vials seeded with Vero cells, and the Gimenez stain technique was used to demonstrate the presence of Rickettsia-like organisms in detached cells found in aspirated medium 19 d after inoculation. These cells were harvested and tested by PCR, targeting portions of the 17-kDa and gltA genes, resulting in products 100% identical to R. felis. This work comprises the first report of R. felis detection in a flea species (A. nudata) endemic to the New World.

Burrows within black-tailed prairie dog (Cynomys ludovicianus) colonies on the Rocky Mountain Arsenal National Wildlife Refuge, Colorado, were dusted with deltamethrin insecticide to reduce flea (Insecta: Siphonaptera) abundance. Flea populations were monitored pre- and posttreatment by combing prairie dogs and collecting fleas from burrows. A single application of deltamethrin significantly reduced populations of the plague vector Oropsylla hirsuta, and other flea species on prairie dogs and in prairie dog burrows for at least 84 d. A plague epizootic on the Rocky Mountain Arsenal National Wildlife Refuge caused high mortality of prairie dogs on some untreated colonies, but did not appear to affect nearby colonies dusted with deltamethrin.

We developed a multiplex polymerase chain reaction (PCR) assay that simultaneously detects three types of flea-associated microorganisms. Targets for the assay were sequences encoding portions of the gltA, a 17-kDa antigen, and pla genes of Bartonella spp. Strong et al., Rickettsia spp. da Rocha-Lima, and Yersinia pestis Yersin, respectively. A total of 260 flea samples containing bloodmeal remnants were analyzed from fleas collected from abandoned prairie dog (Cynomys ludovicianus) burrows at the site of an active plague epizootic in Jefferson County, CO. Results indicated that 34 (13.1%) fleas were positive for Bartonella spp., 0 (0%) were positive for Rickettsia spp., and 120 (46.2%) were positive for Y. pestis. Twenty-three (8.8%) of these fleas were coinfected with Bartonella spp. and Y. pestis. A second group of 295 bloodmeal-containing fleas was collected and analyzed from abandoned burrows in Logan County, CO, where a prairie dog die-off had occurred 2-4 mo before the time of sampling. Of these 295 fleas, 7 (2.4%) were positive for Bartonella spp., 0 (0%) were positive for Rickettsia spp., and 46 (15.6%) were positive for Y. pestis. Coinfections were not observed in fleas from the Logan County epizootic site. The multiplex PCR also was used to identify Y. pestis and Bartonella in prairie dog blood and tissues. This report represents the first identification of Bartonella from prairie dogs and their fleas. Prairie dog fleas were tested with PCR, and the Bartonella PCR amplicons produced were sequenced and found to be closely related to similar sequences amplified from Bartonella that had been isolated from prairie dog blood samples. Phylogenetic analyses indicate that the sequences of bartonellae from prairie dogs and prairie dog fleas cluster tightly within a clade that is distinct from those containing other known Bartonella genotypes.

This study describes the identification of Borrelia parkeri spirochetes in Colorado. Two isolates of B. parkeri (6230 and 6232) were recovered from Ornithodoros parkeri Cooley ticks collected at an inactive prairie dog town in Moffat County. Both isolates were partially characterized by sequencing and subsequent parsimony and neighbor-joining analyses of appropriate regions of the 16S ribosomal RNA, flagellin and P66 genes. Analyses of the 16S gene sequences from the Colorado isolates indicated that they were more closely related to B. parkeri and B. tucatae than to B. hermsii or the other species of Borrelia investigated in this study. Additional analyses of amino acid sequences for flagellin and P66, however, clearly demonstrated that isolates 6230 and 6232 were most closely related to B. parkeri. The possible significance of B. parkeri as an agent of human disease is discussed.

We used a quantitative competitive polymerase chain reaction (PCR) (QC-PCR) to determine bacterial loads in 669 fleas collected in areas of confirmed and suspected plague epizootics. Fleas were collected out of rodent burrows (67.9%) and off of captured animals (24.1%) and rodent carcasses (8.1%). An initial PCR screening assay indicated that 12.1% (81/669) of all fleas were positive for Yersinia pestis. Fleas collected from burrows had significantly lower (chi2 = 264.9, P < 0.0001) infection rates (6.8%) but significantly higher (Student t-test, P < 0.0001) bacterial loads (mean = 10(5.6) Y. pestis per flea) than fleas collected off of rodent carcasses (infection rate = 92.6%; mean bacterial load = 10(4.8) Y. pestis per flea). None of the fleas collected off of captured animals were positive for Y. pestis by PCR, although seven of the 176 captured animals were serologically positive for Y. pestis.

The efficacy of a liquid permethrin-treated bait tube controlling fleas and ticks on Mexican wood rats, Neotoma mexicana Baird, was evaluated during a 1-yr study in north-central Colorado. Results indicated that the bait tubes were effective for reducing flea and tick infestations on wood rats. The effects of treatment persisted throughout the study, despite the fact that bait tubes were replenished with bait and permethrin only during the first 4 mo (4 replenishments). Our results suggest that these bait tubes provide an effective, economical, and environmentally acceptable means of controlling vectors of flea or tick-borne diseases, although slight modifications of the basic bait tube design might be required to maintain the effectiveness of the tube under different ecological conditions.

Eight species of small mammals were evaluated as potential hosts for American dog ticks, Dermacentor variabilis (Say), in an upland, tallgrass prairie study site in central Oklahoma. Only hispid cotton rats, Sigmodon hispidus, and deer mice, Peromyscus maniculatus, were found to be important hosts for immature D. variabilis. Although D. varibilis larvae and nymphs frequently infested both cotton rats and deer mice, cotton rats were the most important host species for both immature stages in the study area. Cotton rats constituted 63.2% of the total 530 small mammals captured and were hosts to 85.2% of all larvae and 88.7% of all nymphs. Deer mice accounted for 19.8% of all small mammals captured and were hosts for 14.5% of the larvae and 10.8% of the nymphs recovered. The remaining small mammal species were hosts for less than 1% of the immature ticks collected. Larval infestations peaked during summer, whereas summer and spring peaks were noted for the nymphal infestations. The relative importance of cotton rats and deer mice as hosts for immature ticks could be largely, but not completely, explained by cotton rats being more than three times as abundant as deer mice. Attachment site data indicated that differences in grooming behavior also might be partially responsible for the larger infestations observed on cotton rats. Other possible ecological and behavioral explanations of the heavy infestations observed on cotton rats are discussed.

Hispid cotton rats (Sigmodon hispidus Say and Ord) were susceptible to infection with Rickettsia rickettsii Wolbach under laboratory conditions and were capable of serving as sources for infecting ticks with rickettsiae. Cotton rats developed rickettsemias that could be detected for as long as 6 h following intraperitoneal inoculation of 10(5) plaque-forming units (PFUs) of R. rickettsii (Morgan strain). An estimate of the median infectious dose (ID50) was made by inoculating six groups (eight animals per group) with serial dilutions of 10(1) to 10(6) PFUs. In this experiment, cotton rats became infected after exposure to relatively few organisms (ID50 = 37 PFUs). None of the 48 cotton rats succumbed to infection, and only 6 of the 48 rats failed to seroconvert. Dermacentor variabilis (Say) larvae were fed on rickettsemic cotton rats to determine whether this species could serve as a source for infecting ticks with R. rickettsii. A small percentage (0.9-3.0%) of larval ticks that fed on three of the four cotton rats had R. rickettsii in their tissues when examined as nymphs. A fourth cotton rat died 7 d after inoculation with R. rickettsii and infected a much higher percentage of ticks (64.0%) than the other three animals.