Demographic features of Bartonella infections in Richardson's ground squirrels (Spermophilus richardsonii)

The epidemiology of Bartonella infections in Richardson's ground squirrels (Spermophilus richardsonii) was studied at multiple sites in Saskatchewan, Canada, from 2002 to 2004. The overall prevalence of Bartonella infection was 48%. Juvenile squirrels were significantly more likely to be infected with Bartonella than were adults (58% and 37%, respectively), and juvenile animals also were significantly more likely to have high levels of bacteremia compared to adult animals. Prevalence of Bartonella infection appeared to decrease with age; only 24% of animals known to be > or = 2 yr old were infected with Bartonella. Prevalence of infection was lowest in May (27%) and highest in late summer and early autumn (71%). The prevalence of fleas also varied seasonally, and animals were more likely to have fleas in the late summer and early autumn than in early summer. We found no relationship between Bartonella prevalence and host density or flea prevalence.     

Ectoparasites of gray squirrels in two different habitats and screening of selected ectoparasites for bartonellae

Gray squirrels, Sciurus carolinensis, were livetrapped in 2 different habitat types, woodland (67 squirrels) and parkland (53 squirrels), in southeastern Georgia. Ectoparasites were recovered from anesthetized squirrels and compared between hosts from the 2 habitats. Because of the absence of low vegetation in parkland habitats, it was hypothesized that the ectoparasite fauna, especially ticks and chiggers, would be more diverse on woodland squirrels. The results were generally in agreement with this hypothesis. Seventeen species of ectoparasites were recovered from woodland squirrels, compared with 6 species from parkland squirrels. Five species of ticks and 3 species of chiggers parasitized the woodland squirrels compared with no ticks or chiggers on the parkland squirrels. Significantly higher infestation prevalences were recorded on woodland compared with parkland squirrels for the flea Orchopeas howardi, the tick Amblyomma americanum, and the mesostigmatid mite Androlaelaps fahrenholzi. The mean intensity for O. howardi also was significantly higher on woodland than on parkland squirrels. Because a new strain of Bartonella sp. was isolated recently from S. carolinensis in Georgia, selected ectoparasites from this study were screened for bartonellae by polymerase chain reaction (PCR). Some of the fleas and lice, but none of the mites tested, were PCR positive, suggesting that fleas, or lice, or both, might be vectors of bartonellae between squirrels. Six distinct strains of Bartonella sp. were detected, 2 in fleas and 4 in lice.     

Parasite removal and its impact on litter size and body condition in Columbian ground squirrels (Spermophilus columbianus)

The current interest in studies of parasite-host relationships is focusing on the impact of parasites on mate choice, sexual selection and individual fitness. While most studies have been done on birds and fishes, little is known about the interactions between parasites and reproductive success in mammals. In this study, I experimentally removed ectoparasites (mainly fleas) from some female Columbian ground squirrels using a commercially available powder, whereas other squirrels were left untreated as controls. The anti-parasite treatment led to an increase in female body condition during lactation and at weaning and an increase in litter size. The increase of body mass in treated females between birth of the young and weaning is also different from observations done over 8 years in the same population, where untreated females usually lost mass. In close to 400 litters from untreated females in the same area, only 2% had litter sizes as big as the treated females. I conclude that ectoparasites can have a profound impact not only on individuals but probably on the dynamics of the whole population.     

Phylogenetic analysis of Bartonella detected in rodent fleas in Yunnan, China

Previous studies have demonstrated a diversity of Bartonella spp. in rodent populations in Yunnan Province, China. Although Bartonella spp. have been isolated from cat fleas and cattle ticks collected from their animal hosts, little is known about Bartonella carried by rodent fleas. In this study, Bartonella DNA was detected by polymerase chain reaction (PCR) in two of five species of rodent fleas. These included Xenopsylla cheopis and Ctenophthalmus lushuiensis, which were collected from Rattus tanezumi flavipectus and from the nests of voles, respectively, during 1997 from two sites in western Yunnan Province, China. Sequence analysis of the Bartonella citrate synthase gene (gltA) amplicons obtained from six of 65 grouped flea samples showed that Bartonella genetic variants were clustered in four groups. One from Xenopsylla cheopis was identical to Bartonella tribocorum, whereas the other three genotypes from Ctenophthalmus lushuiensis were related to the vole-associated Bartonella isolates and cat-associated Bartonella clarridgeiae. This is the first detection of this Bartonella variant from fleas in China. Therefore, further investigations are needed to clarify the distribution of Bartonella in rodents and their ectoparasites in China to define the role of these arthropods in the transmission routes of Bartonella.     

The "hibernation induction trigger": specificity and validity of bioassay using the 13-lined ground squirrel

Even though the existence of the blood-borne "hibernation induction trigger" has been reported in the 13-lined ground squirrel, transfusion of plasma from hibernating rodents with other hibernating species as the recipients failed to induce the occurrence of summer hibernation. In order to verify whether the response to the "trigger" substance is species specific, the present study was carried out to compare the effect of plasma from hibernating Richardson's ground squirrels on the incidence of summer hibernation in both juvenile Richardson's and adult 13-lined ground squirrels. In two series of experiments, 13-lined ground squirrels entered hibernation quite readily independent of the treatment. The rate of occurrence of hibernation ranged from 78% after sham injection to 86% after warm saline, fresh summer active plasma, and fresh hibernating plasma, respectively. There were no differences in the number of hibernation bouts and the number of days in hibernation after each treatment. In contrast, none of the juvenile Richardson's ground squirrels entered hibernation after any of the treatments up to the end of the 8-week observation period. These results not only argue against the existence of blood-borne "trigger" substance, at least in the Richardson's ground squirrel, but also caution against the use of the 13-lined ground squirrel as a standard test animal for the bioassay of the "trigger" substance.     

Donning your enemy's cloak: ground squirrels exploit rattlesnake scent to reduce predation risk

Ground squirrels (Spermophilus spp.) have evolved a battery of defences against the rattlesnakes (Crotalus spp.) that have preyed on them for millions of years. The distinctive behavioural reactions by these squirrels to rattlesnakes have recently been shown to include self-application of rattlesnake scent-squirrels apply scent by vigorously licking their fur after chewing on shed rattlesnake skins. Here, we present evidence that this behaviour is a novel antipredator defence founded on exploitation of a foreign scent. We tested three functional hypotheses for snake scent application--antipredator, conspecific deterrence and ectoparasite defence--by examining reactions to rattlesnake scent by rattlesnakes, ground squirrels and ectoparasites (fleas). Rattlesnakes were more attracted to ground squirrel scent than to ground squirrel scent mixed with rattlesnake scent or rattlesnake scent alone. However, ground squirrel behaviour and flea host choice were not affected by rattlesnake scent. Thus, ground squirrels can reduce the risk of rattlesnake predation by applying rattlesnake scent to their bodies, potentially as a form of olfactory camouflage. Opportunistic exploitation of heterospecific scents may be widespread; many species self-apply foreign odours, but few such cases have been demonstrated to serve in antipredator defence.     

Rodent-flea-plague relationships at the higher elevations of San Diego County, California.

Rodent-flea-plague relationships were examined at sites located at higher elevations of San Diego County during 1991-2002. The most frequently sampled rodents were the California ground squirrel, Spermophilus beecheyi, and white-footed mice, Peromyscus spp. Higher seasonal mean dependent variables (prevalences of squirrels and indices of O. montana), higher mean amounts of seasonal rainfall, and lower seasonal mean ambient temperatures were found at sites during 1991--1993 compared with 2000--2002. Levels of plague antibody indicate that squirrels seroconverted at most sites during spring/early summer when exposed to infected fleas when cleaning out burrows following hibernation, when interacting more with other squirrels occurred establishing home ranges, and when dispersing juveniles occupied abandoned burrows containing old nests. The majority of squirrels (35/41) were reported seropositive at Heise County Park (elev. 1,281 m) during 1996, 2001, and 2002. This may reflect fewer fleas biting squirrels more often when squirrels spent more time below-ground in order to avoid higher ambient temperatures. In contrast, most squirrels (12/13) were reported seropositive at two higher sites (elev. 1,584 and 1,641 m) on Palomar Mountain during 1993--1996 when there were higher mean dependent variables and amounts of seasonal rainfall compared with 1999--2002. This increased rainfall may have benefited host and flea populations and thus plague transmission. Seasonal indices were usually reduced after dusting squirrel burrows with 2% diazinon dust. Findings also indicated that S. beecheyi is a primary reservoir for plague at the higher elevations of these and other sites in southern California, and that the deer mouse, Peromyscus maniculatus, and other Peromyscus spp. were not major host reservoirs at the sites sampled.     

Molecular detection and identification of Bartonella species in Xenopsylla cheopis fleas (Siphonaptera: Pulicidae) collected from Rattus norvegicus rats in Los Angeles, California

Of 200 individual Xenopsylla cheopis fleas removed from Rattus norvegicus rats trapped in downtown Los Angeles, CA, 190 (95%) were positive for the presence of Bartonella DNA. Ninety-one amplicons were sequenced: Bartonella rochalimae-like DNA was detected in 66 examined fleas, and Bartonella tribocorum-like DNA was identified in 25 fleas. The data obtained from this study demonstrate an extremely high prevalence of Bartonella DNA in rat-associated fleas.     

Managing plague on prairie dog colonies: insecticides as ectoparasiticides

Human health practitioners and wildlife biologists use insecticides to manage plague by suppressing fleas (Siphonaptera), but insecticides can also kill other ectoparasites. We investigated effects of deltamethrin and fipronil on ectoparasites from black‐tailed prairie dogs (Cynomys ludovicianus, BTPDs). In late July, 2018, we treated three sites with 0.05% deltamethrin dust and 5 sites with host‐fed 0.005% fipronil grain. Three non‐treated sites functioned as experimental baselines. We collected ectoparasites before treatments (June‐July, 2018) and after treatments (August‐October, 2018, June‐July, 2019). Both deltamethrin and fipronil suppressed fleas for at least 12 months. Deltamethrin had no detectable effect on mites (Arachnida). Fipronil suppressed mites for at least 12 months. Lice (Phthiraptera) were scarce on non‐treated sites throughout the study, complicating interpretation. Concentrating on eight sites where all three ectoparasites where found in June‐July, 2018 (before treatments), flea intensity was greatest on BTPDs carrying many lice and mites. These three ectoparasites co‐occurred at high numbers, which might facilitate plague transmission in some cases. Lethal effects of insecticides on ectoparasite communities are potentially advantageous in the context of plague management.     

Natural infections of Richardson's ground squirrels with western equine encephalomyelitis virus, Saskatchewan, Canada, 1964-1973.

A survey (1964-1973) was conducted on naturally occurring western equine encephalomyelitis (WEE) virus infections of Richardson's ground squirrels (RGS) in agricultural areas of Saskatchewan. The survey included both serology and virus isolation attempts on live-trapped wild squirrels. Throughout the study, seropostive squirrels were found in the known enzootic regions of the province each summer. The infections rate was high (11.6% of 250) in the epidemic year of 1965 and low (less than 2% of 681) in the subsequent non-epidemic years. The maximal seasonal prevalence of seropositive RGS coincided with the time when the squirrel population and aedine mosquito abundance reached their seasonal peaks. Five virus isolations were obtained from the blood and brains of naturally infected squirrels. All isolations were from squirrels collected in June. The early seasonal infections in squirrels could provide a source of virus for Culex tarsalis, the epidemic vector. In view of their abundance in the enzootic agricultural areas and the high annual population turnover, the RGS may play an important role in the natural history of WEE virus in the Canadian prairies.     

The impact of predation on burrow use by arctic ground squirrels in the boreal forest

In sedentary animals, the choice of a suitable home site is critical to survival and reproductive fitness. However, habitat suitability may vary with predation risk. We compared habitat use of Arctic ground squirrels (Spermophilus parryii plesius) living in the boreal forest under conditions of fluctuating predation pressure. In our study area, predators show ten-year cycles in numbers that track that of their primary prey, the snowshoe hare (Lepus americanus). In 1993, we compared burrows that continued to be occupied following the period of intense predation during the hare decline of 1990-1992 with those that became vacant, and with random locations. We contrasted these sites to those in a predator exclosure where predation pressure was minimized. Burrows on control sites were located on sloped sites with high visibility. Burrows that remained occupied during the period of intense predation were more likely to be in open areas with fewer fallen trees than burrows that became vacant. We used discriminant functions derived from the control sites and found that 89% of the burrows on the predator exclosure were classified as being similar to the random locations on control sites. We conclude that the distribution of Arctic ground squirrels in the boreal forest is a direct function of predator presence.     

Seasonal and among‐site variation in the occurrence and abundance of fleas on California ground squirrels (Otospermophilus beecheyi)

An improved understanding of the ecology of fleas on California ground squirrels, Otospermophilus beecheyi, is warranted given the role of fleas in the transmission, and perhaps persistence, of the plague‐causing bacterium Yersinia pestis. We sampled O. beecheyi on a seasonal basis from three study sites, each representing a different land use type (preserve, pasture, and agriculture) in the San Joaquin Valley, CA. Overall, the abundance of fleas on squirrels was greatest in spring at the preserve site, in summer at the agriculture and pasture sites, and in winter at the pasture site. Hoplopsyllus anomalus, the species most frequently found on squirrels, was most abundant in spring at the preserve site and in summer at the agriculture and pasture sites. Oropsylla montana was most abundant in winter at the pasture site and on adult squirrels. Echidnophaga gallinacea was most abundant in fall on juvenile squirrels at the preserve site. All three flea species we encountered are known to be potential vectors of Y. pestis. Future efforts to predict flea species occurrence and abundance (and plague risk) at sites of concern should consider seasonal microclimatic conditions and the potential influence of human land use practices.     

Acquisition of nonspecific Bartonella strains by the northern grasshopper mouse (Onychomys leucogaster)

Rodent-associated Bartonella species are generally host-specific parasites in North America. Here evidence that Bartonella species can 'jump' between host species is presented. Northern grasshopper mice and other rodents were trapped in the western USA. A study of Bartonella infection in grasshopper mice demonstrated a high prevalence that varied from 25% to 90% by location. Bartonella infection was detected in other rodent species with a high prevalence as well. Sequence analyses of gltA identified 29 Bartonella variants in rodents, 10 of which were obtained from grasshopper mice. Among these 10, only six variants were specific to grasshopper mice, whereas four were identical to variants specific to deer mice or 13-lined ground squirrels. Fourteen of 90 sequenced isolates obtained from grasshopper mice were strains found more commonly in other rodent species and were apparently acquired from these animals. The ecological behavior of grasshopper mice may explain the occurrence of Bartonella strains in occasional hosts. The observed rate at which Bartonella jumps from a donor host species to the grasshopper mouse was directly proportional to a metric of donor host density and to the prevalence of Bartonella in the donor host, and inversely proportional to the same parameters for the grasshopper mouse.     

Sarcocystis campestris from naturally infected 13-lined ground squirrels, Spermophilus tridecemlineatus tridecemlineatus, from Nebraska

Grossly visible sarcocysts were seen in the skeletal muscles of 1 of 12 13-lined ground squirrels, Spermophilus tridecemlineatus tridecemlineatus, collected in Nebraska. The tissue cyst wall was up to 5.0 microm thick and contained spikelike projections. Transmission electron microscopy of tissue cysts revealed they were similar to Sarcocystis campestris Cawthorn, Wobeser, and Gajadhar, 1983, previously known only from experimental infections in Richardson's ground squirrel Spermophilus richardsonii. Prominent electron-dense bodies were observed lining the microfilaments present in the spikelike projections of the sarcocyst wall. This is the first report of S. campestris in a natural intermediate host and the first report of this parasite outside of Saskatoon, Canada.     

The influence of the host on the frequency of block formation in males and females of Citellophilus tesquorum altaicus Ioff, 1936 (Siphonaptera: Ceratophyllidae) and transmission of the plague pathogen by these fleas

Data obtained during feeding of infested females and males of Citellophilus tesquorum altaicus Ioff, 1936 (Siphonaptera: Ceratophyllidae), the main vector of plague in Tuva natural plague focus, on the natural hosts and on laboratory animals were analyzed. Sexual differences in fleas were found to depend on the host. Females fed more actively on longtailed ground squirrels Spermophilus undulatus than on white mice. The feeding activity of males on the two hosts did not differ. A higher mortality of infested females and males was observed during feeding on mice. The frequency of block formation and pathogen transmission in males was higher during bloodsucking on the ground squirrel; in females, during feeding on mice. Thus, sex-related differences in the plague pathogen transmission revealed in laboratory white mice may be not the same in nature. This should be taken into account when extrapolating the experimental data on the natural relations between the plague pathogen and ectoparasites.     

Mixed infections, cryptic diversity, and vector-borne pathogens: evidence from Polygenis fleas and Bartonella species

Coinfections within hosts present opportunities for horizontal gene transfer between strains and competitive interactions between genotypes and thus can be a critical element of the lifestyles of pathogens. Bartonella spp. are Alphaproteobacteria that parasitize mammalian erythrocytes and endothelial cells. Their vectors are thought to be various biting arthropods, such as fleas, ticks, mites, and lice, and they are commonly cited as agents of various emerging diseases. Coinfections by different Bartonella strains and species can be common in mammals, but little is known about specificity and coinfections in arthropod vectors. We surveyed the rate of mixed infections of Bartonella in flea vectors (Polygenis gwyni) parasitizing cotton rats (Sigmodon hispidus) in which previous surveys indicated high rates of infection. We found that nearly all fleas (20 of 21) harbored one or more strains of Bartonella, with rates of coinfection approaching 90%. A strain previously identified as common in cotton rats was also common in their fleas. However, another common strain in cotton rats was absent from P. gwyni, while a rare cotton rat strain was quite common in P. gwyni. Surprisingly, some samples were also coinfected with a strain phylogenetically related to Bartonella clarridgeiae, which is typically associated with felids and ruminants. Finally, a locus (pap31) that is characteristically borne on phage in Bartonella was successfully sequenced from most samples. However, sequence diversity in pap31 was novel in the P. gwyni samples, relative to other Bartonella previously typed with pap31, emphasizing the likelihood of large reservoirs of cryptic diversity in natural populations of the pathogen.     

Burrow Use and the Influence of Ectoparasites in Brants' Whistling Rat Parotomys brantsii

In Brants' whistling rat, Parotomys brantsii, single animals inhabit unusually complex burrow systems containing several nest chambers. We investigated burrow use and examined the effect of ectoparasites on choice of alternative nest chambers by radiotelemetrically monitoring the nocturnal sleeping locations of individual whistling rats, before and after treating them to remove ectoparasites. Prior to treatment, animals used several different nest chambers within a single burrow, moving from one chamber to another every 1.6 d on average, and most individuals also slept in more than one burrow. Anti‐parasite treatment reduced the rate at which animals switched from one nest chamber to another within a burrow. Screening of a separate sample of animals for ectoparasites revealed that they were infested with fleas, mainly Xenopsylla eridos. We suggest that by switching periodically from one nest chamber to another, whistling rats reduce the rate at which ectoparasites, especially fleas, accumulate.     

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.     

Bartonella and Rickettsia from fleas (Siphonaptera: Ceratophyllidae) of prairie dogs (Cynomys spp.) from the western United States

Fleas of prairie dogs have been implicated in the transmission of Bartonella spp. We used PCR to test DNA extracts from 47 fleas of prairie dogs from 6 states. We amplified DNA from 5 unique genotypes of Bartonella spp. and 1 Rickettsia sp. from 12 fleas collected in North Dakota, Oklahoma, Texas, and Wyoming. Sequences from the Bartonella spp. were similar, but not identical, to those from prairie dogs and their fleas in Colorado.     

The shape of things eaten: the functional response of herbivores foraging adaptively

We studied reproductive and somatic investments in >700 female Richardson's ground squirrels ( Spermophilus richardsonii ) of known age over a 14-year period to evaluate three hypotheses, restraint, senescence, and residual reproductive value, proposed to explain age-specific life history patterns in iteroparous vertebrates. We found that reproductive investment, measured as litter mass at first emergence from the natal nest, did not differ among age classes. Although yearling female Richardson's ground squirrels made a greater somatic investment during reproduction than older females, they produced similar numbers and mass of offspring as older females. Reproductive investment did not decline with age, though active season somatic investment was lowest in the oldest females. Somatic investment during reproduction was highest in yearlings. This combination of age-related changes in somatic investment unaccompanied by changes in reproductive investment was not well explained by any of the hypotheses examined, though the senescence hypothesis best explained the combination of declining somatic investment and declining survival of the oldest females. Our results supported the ideas that reproductive and somatic senescence evolve independently, and that somatic senescence may be more common in relatively smaller species.