Infected Nymphs Research Articles

Effects of the infection with Trypanosoma cruzi on the feeding and excretion/defecation patterns of Triatoma infestans.

Transmission of Trypanosma cruzi (Kinetoplastida: Trypanosomatidae) occurs when feces/urine of infected triatomines come into contact with mucous membranes or damaged skin, and this occurs mainly when insects defecate while feeding on the host. Thus, the vector competence of the triatomines is associated with their feeding and excretion/defecation behavior. This work studied for the first time the effect of T. cruzi infection on feeding and excretion/defecation patterns of Triatoma infestans (Hemiptera: Reduviidae). Uninfected and infected fifth-instar nymphs were fed ad libitum and their feeding behavior and defecations were registered during and after feeding. The feeding pattern did not show differences between the experimental groups. However, the infected nymphs began to defecate earlier, defecated in greater quantity and there was a greater proportion of defecating individuals compared to uninfected nymphs. These results show that T. cruzi affected the excretion/defecation pattern of T. infestans in a way that would increase the probability of contact between infective feces and the mammalian host.

Influence of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae) Infection on Mortality of the Sylvatic Triatomine Vector, Mepraia spinolai (Heteroptera: Reuviidae), Under Fasting.

The etiologic agent of Chagas disease, Trypanosoma cruzi, is transmitted by hematophagous insect vectors that subsist on repeated blood meals over their lives separated by periods of fasting. Using naturally infected Mepraia spinolai, we measured the influence of parasite infection on this host vector's mortality during regular feeding and after fasting. After their capture, the insects were fed twice with uninfected mice to evaluate parasitic infection in their fecal samples by microscopic observation and PCR. Then the insects were subjected to a fasting period, followed by a third (final) feeding. After each feeding, a fecal sample was obtained to evaluate T. cruzi infection. To determine its progress through ontogeny, mortality and ecdysis of the infected and uninfected nymphs and adults were recorded on three occasions, over 140 d, and analyzed. Detections of infection by T. cruzi between the two first feedings increased, but this detection level was generally reduced after final feeding unless reinfected. For nymphs (stages III-V), their mortality was highest when infected after the fasting period, whereas adults were equally resistant to death after fasting when infected with T. cruzi. Metacyclic trypomastigotes were principally excreted in the fecal samples. Our results confirm that T. cruzi is pathogenic to its invertebrate hosts under nutritional stress conditions, when nymphs' mortality is higher while infected than uninfected when they were hungry. These results are epidemiologically important because T. cruzi harms the fasting vector M. spinolai, reducing its lifespan and competence as a disease vector, and thereby its rates of parasite transmission.

Lyme neuroborreliosis and bird populations in northern Europe.

Many vector-borne diseases are transmitted through complex pathogen-vector-host networks, which makes it challenging to identify the role of specific host groups in disease emergence. Lyme borreliosis in humans is now the most common vector-borne zoonosis in the Northern Hemisphere. The disease is caused by multiple genospecies of Borrelia burgdorferi sensu lato bacteria transmitted by ixodid (hard) ticks, and the major host groups transmit Borrelia genospecies with different pathogenicity, causing variable clinical symptoms in humans. The health impact of a given host group is a function of the number of ticks it infects as well as the pathogenicity of the genospecies it carries. Borrelia afzelii, with mainly small mammals as reservoirs, is the most common pathogen causing Lyme borreliosis, and it is often responsible for the largest proportion of infected host-seeking tick nymphs in Europe. The bird-borne Borrelia garinii, though less prevalent in nymphal ticks, is more likely to cause Lyme neuroborreliosis, but whether B. garinii causes disseminated disease more frequently has not been documented. Based on extensive data of annual disease incidence across Norway from 1995 to 2017, we show here that 69% of disseminated Lyme borreliosis cases were neuroborreliosis, which is three times higher than predicted from the infection prevalence of B. garinii in host-seeking ticks (21%). The population estimate of migratory birds, mainly of thrushes, explained part of the annual variation in cases of neuroborreliosis, with a one-year time lag. We highlight the important role of the genospecies' pathogenicity and the host associations for understanding the epidemiology of disseminated Lyme borreliosis.

Predicting spatiotemporal patterns of Lyme disease incidence from passively collected surveillance data for Borrelia burgdorferi sensu lato-infected Ixodes scapularis ticks

Lyme disease is the most prevalent vector-borne disease in the United States. Ixodes scapularis, commonly referred to as the blacklegged tick, is the primary vector of Lyme disease spirochetes, Borrelia burgdorferi sensu lato (s.l.), in the eastern United States. Connecticut has pervasive populations of I. scapularis and remains a hotspot for Lyme disease. A primary aim of this study was to determine if passively collected data on human-biting I. scapularis ticks in Connecticut could serve as a useful proxy for Lyme disease incidence based on the cases reported by the Connecticut Department of Public Health (CDPH). Data for human-biting I. scapularis ticks submitted to the Tick Testing Laboratory at the Connecticut Agricultural Experiment Station (CAES-TTL), and tested for infection with B. burgdorferi s.l., were used to estimate the rate of submitted nymphs, nymphal infection prevalence, and the rate of submitted infected nymphs. We assessed spatiotemporal patterns in tick-based measures and Lyme disease incidence with generalized linear and spatial models. In conjunction with land cover and household income data, we used generalized linear mixed effects models to examine the association between tick-based risk estimates and Lyme disease incidence. Between 2007 and 2017, the CAES-TTL received 26,116 I. scapularis tick submissions and the CDPH reported 23,423 Lyme disease cases. The rate of submitted nymphs, nymphal infection prevalence, the rate of submitted infected nymphs, and Lyme disease incidence all decreased over time during this eleven-year period. The rate of submitted nymphs, the rate of submitted infected nymphs, and Lyme disease incidence were spatially correlated, but nymphal infection prevalence was not. Using a mixed modeling approach to predict Lyme disease incidence and account for spatiotemporal structuring of the data, we found the best fitting tested model included a strong, positive association with the rate of submitted infected nymphs and a negative association with the percent of developed land for each county. We show that within counties, submissions of B. burgdorferi s.l. infected nymphs were strongly and positively associated with inter-annual variation in reported Lyme disease cases. Tick-based passive surveillance programs may be useful in providing independent measures of entomological risk, particularly in settings where Lyme disease case reporting practices change substantially over time.

Behavioral thermoregulation in Locusta migratoria manilensis (Orthoptera: Acrididae) in response to the entomopathogenic fungus, Beauveria bassiana.

Insects such as locusts and grasshoppers can reduce the effectiveness of pathogens and parasites by adopting different defense strategies. We investigated the behavioral thermopreference of Locusta migratoria manilensis (Meyen) (Orthoptera: Acrididae) induced by the fungus Beauveria bassiana, and the impact this behavior had on the fungal mycosis under laboratory conditions. By basking in higher temperature locations, infected nymphs elevated their thoracic temperature to 30–32.6 °C, which is higher than the optimum temperature (25°C) for B. bassiana conidial germination and hyphal development. A minimum thermoregulation period of 3 h/day increased survival of infected locusts by 43.34%. The therapeutic effect decreased when thermoregulation was delayed after initial infection. The fungus grew and overcame the locusts as soon as the thermoregulation was interrupted, indicating that thermoregulation helped the insects to cope with infection but did not completely rid them of the fungus. A significant enhancement in the number of haemocytes was observed in infected thermoregulating locusts, reaching levels that were even higher than those observed in the controls. In contrast, haemocyte concentration was severely reduced in infected insects that did not thermoregulate. In infected non-thermoregulating locusts, the reduction in haemocyte number was accompanied by an increase in fungal blastospore concentration that was obvious in the haemolymph by day four. In contrast, no circulating blastospores were found in the haemolymph of infected thermoregulating locusts three days post-inoculation. We also examined the phagocytic activity of infected insects in vivo by using fluorescein isothiocyanate (FITC)-labelled silica beads. The proportion of beads that was engulfed by haemocytes in infected, thermoregulating insects was similar to that in the controls throughout the experiment, whereas the rate of phagocytosis in infected, non-thermoregulating insects progressively decreased after infection. These findings demonstrated that behavioural thermoregulation can adversely affect B. bassiana mycosis in infected L. migratoria manilensis, thereby limiting the development of lethal entomopathogenic fungi in locusts. This is apparently accomplished through an increase in the levels of haemocytes, leading to greater phagocytic activity under certain environmental conditions.

Rickettsia rickettsii Co-feeding Transmission among Amblyomma aureolatum Ticks.

Amblyomma aureolatum ticks are vectors of Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever in Brazil. Maintenance of R. rickettsii in nature depends on horizontal transmission along tick generations. Although such transmission is known to occur when uninfected and infected ticks feed simultaneously on susceptible animals (co-feeding systemic transmission), we investigated co-feeding nonsystemic transmission, which was based on R. rickettsii–infected and –uninfected A. aureolatum ticks feeding simultaneously on guinea pigs immune to R. rickettsii. Our acquisition and transmission infestations demonstrated that horizontal transmission of R. rickettsii by co-feeding ticks on immune hosts with no systemic infection did not occur when uninfected larvae fed distantly from infected nymphs but did occur in a few cases when uninfected larvae fed side-by-side with infected nymphs, suggesting that they shared the same feeding site. The co-feeding nonsystemic transmission type might have no epidemiologic importance for Rocky Mountain spotted fever.

SUSCEPTIBILITY OF DESERT LOCUST, SCHISTOCERCA GREGARIA (ORTHOPTERA: ACRIDIDAE) TO BACILLUS CEREUS ISOLATED FROM EGYPT

Examination was done at preliminary bracketing bioassay on one old 4th nymphal instar of desert locust. Results showed that two isolates, namely NDL1 and NDL2 were having highly potentiality as entomopathogenic bioagents. Thirty isolates were isolated from dead/ infected nymphs of desert locust occurred in raring cages at Department of Locust and Grasshoppers Research, Plant Protection Research Institute, Agricultural Research Center, Dokki, Giza, Egypt. Molecular identification of isolated bacteria was done using universal primers of 16s rRNA, followed by DNA sequencing. Nucleotides were blasted at (https://www.ncbi. nlm.nih.gov /genbank/) to recognize that NDL1 and NDL2 isolates were two different isolates of Bacillus cereus with a high similarity (100%). Susceptibility of 4th nymphal instar of Schistocerca gregaria (Forskal) to the isolated B. cereus was determined using two bioassay procedures, Leaf-dip and per os. The insecticidal activity of both isolates against locust nymph in leaf dipping showed that NDL2 was more efficient than NDL1. However, the opposite trend was observed in using per os. Both Isolates have the potential to be a successful biocidal agent to control desert locust.

Effects of Rickettsia amblyommatis Infection on the Vector Competence of Amblyomma americanum Ticks for Rickettsia rickettsii.

Although Dermacentor spp. ticks are considered the primary vectors of Rickettsia rickettsii in the United States, other North American tick species are also capable of transmitting the agent, including the lone star tick-Amblyomma americanum. The lone star tick is an aggressive human-biting tick abundant in the South, Central, and Mid-Atlantic United States, which has been shown to be a competent vector of R. rickettsii in laboratory studies. However in nature, A. americanum frequently carry Rickettsia amblyommatis-another member of the spotted fever group-with the prevalence of infection reaching 84% in some populations. It has been postulated that the presence of an endosymbiotic Rickettsia in a significant proportion of a vector population would diminish or even block transmission of pathogenic Rickettsia in ticks from generation to generation due to transovarial interference. We measured the ability of R. amblyommatis-infected A. americanum to acquire R. rickettsii from an infected host with a bloodmeal, and transmit it transstadially, horizontally (to a susceptible host), and vertically to the next generation. Larvae from both the R. amblyommatis-infected and R. amblyommatis-free cohorts acquired R. rickettsii from infected guinea pigs, but the presence of the symbiont diminished the ability of coinfected engorged larvae to transmit R. rickettsii transstadially. Conversely, acquisition of R. rickettsii by cofeeding was unaffected in R. amblyommatis-infected nymphs and adults; prevalence of R. rickettsii in engorged adults reached 97% in both R. amblyommatis-infected and R. amblyommatis-free cohorts. In guinea pigs exposed to dually infected nymphs, R. rickettsii infection was milder than in those fed upon nymphs infected with R. rickettsii only. The frequency of transovarial transmission of R. rickettsii in the R. amblyommatis-infected cohort (31%) appeared lower than that in the R. amblyommatis-free cohort (48%), but the difference was not statistically significant. Larval progenies of dually infected A. americanum females transmitted R. rickettsii to naïve guinea pigs confirming viability of the pathogen. Thus, the vector competence of A. americanum for R. rickettsii was not significantly affected by R. amblyommatis.

Kyasanur forest disease: a rare viral hemorrhagic disease in India

Kyasanur forest disease (KFD) is a rare hemorrhagic disease in India and isolated in monkeys in the Kyasanur Forest of the Shimoga district, Karnataka State in India in 1957. KFD is a zoonotic disease and endemic in southern part of India. KFD Virus commonly infects the black faced langur monkey (Semnopithecus entellus). No evidence for human-to-human transmission. Large domestic animals (cows, goats, sheep) that become infected are thought be important only for sustaining tick population. When monkeys come in contact with the infected ticks, they get infected, amplify and disseminate the infection creating hot spots of infection. The people who pass through the forest are bitten by the infected nymphs of H. spinigera, which are highly anthropophilic. Local villagers staying in and around the forest area frequently visit the forest for collection of fire woods, grass and get infected through tick bites. The incubation period is 3-8 days and patients presented with following symptoms like chills, frontal headache, bodyache, and high fever for 5-12 days. There were 466 human cases during initial outbreak and 181 more the following year and has caused epidemic outbreaks of haemorrhagic fever affecting 400 to 1000 people per year since then, with mortality rate of 4 and 15%. In 2012, 9 persons were found positive for KFD, in Theerthahlli taluk of district Karnataka. There is no specific treatment for KFD but a timely supportive therapy reduces the mortality in human being.

Increase of Albinistic Hosts Caused by Gut Parasites Promotes Self-Transmission.

Paranosema locustae is a gut parasite that has been applied widely in the control of grasshoppers in many parts of the world. Usually, P. locustae is transmitted horizontally via passive modes under natural conditions but in the current study, a positive transmission strategy of P. locustae was demonstrated. First, infection by P. locustae resulted in the cuticula of infected Locusta migratoria nymphs to become lighter in color: normally only a small proportion of locusts are pale with most either being partly or mostly black; but locusts infected with P. locustae became pale. And it was found that the change to pale occurred even among uninfected black and partly black nymphs reared with infected locusts. The eumelanin of the thorax and abdomen of infected individuals decreased significantly, as did the level of dopamine. In addition, there was a decrease in phenol oxidase activity and the expression of henna and pale, which are involved in the synthesis of cuticle melanin, decreased. What is the ecological significance of this increase in light-colored hosts caused by P. locustae? We discovered that light-colored locusts were more susceptible to the microsporidian pathogen than dark-colored individuals were, because of their weaker melanization. Phenol oxidase activity in pale locusts was lower than that of black locusts, but the serpin expression level of pale locusts was higher than that of black individuals. When examined for infection, it was found that initially uninfected nymphs had picked up P. locustae infections indicating that infections are readily passed from one pale locust to another. The infection rate of healthy locusts reared with light-colored locusts infected with P. locustae was 100% which was more than with black-colored ones. The increase in albinistic locusts clearly promoted the prevalence of P. locustae in the total population. In conclusion, these results elucidated a new strategy of positive self-transmission in P. locustae.Importance:Mother Nature always grants wisdom to her creatures and feeds them carefully. This wisdom is particularly apparent in the relationships between two interacting species. In this study, our team focused on the interaction between L. migratoria and P. locustae. In a previous study, it was found that L. migratoria isolate infected individuals, reducing avoiding the spread of P. locustae, in a previous study. The solitary, pale individuals infected by P. locustae were left behind as locust groups marched ahead, leading to a kind of behavioral immunity in the insects. Here, we reported that P. locustae promotes pigmentation loss in L. migratoria, causing a larger proportion of light-colored individuals, and these lighter individuals which possessed weaker immunity against pathogens. This strategy is advantageous to P. locustae, as it promotes its propagation and spread. These extraordinary abilities of L. migratoria and P. locustae have accumulated over millennia of years of interaction.

Tick-borne disease risk in a forest food web.

Changes to the community ecology of hosts for zoonotic pathogens, particularly rodents, are likely to influence the emergence and prevalence of zoonotic diseases worldwide. However, the complex interactions between abiotic factors, pathogens, vectors, hosts, and both food resources and predators of hosts are difficult to disentangle. Here we (1) use 19yr of data from six large field plots in southeastern New York to compare the effects of hypothesized drivers of interannual variation in Lyme disease risk, including the abundance of acorns, rodents, and deer, as well as a series of climate variables; and (2) employ landscape epidemiology to explore how variation in predator community structure and forest cover influences spatial variation in the infection prevalence of ticks for the Lyme disease bacterium, Borrelia burgdorferi, and two other important tick-borne pathogens, Anaplasma phagocytophilum and Babesia microti. Acorn-driven increases in the abundance of mice were correlated with a lagged increase in the abundance of questing nymph-stage Ixodes scapularis ticks infected with Lyme disease bacteria. Abundance of white-tailed deer 2 yr prior also correlated with increased density of infected nymphal ticks, although the effect was weak. Density of rodents in the current year was a strong negative predictor of nymph density, apparently because high current abundance of these hosts can remove nymphs from the host-seeking population. Warm, dry spring or winter weather was associated with reduced density of infected nymphs. At the landscape scale, the presence of functionally diverse predator communities or of bobcats, the only obligate carnivore, was associated with reduced infection prevalence of I.scapularis nymphs with all three zoonotic pathogens. In the case of Lyme disease, infection prevalence increased where coyotes were present but smaller predators were displaced or otherwise absent. For all pathogens, infection prevalence was lowest when forest cover within a 1km radius was high. Taken together, our results suggest that a food web perspective including bottom-up and top-down forcing is needed to understand drivers of tick-borne disease risk, a result that may also apply to other rodent-borne zoonoses. Prevention of exposure based on ecological indicators of heightened risk should help protect public health.

Occurrence of Purpureocillium lilacinum in citrus black fly nymphs

Abstract Black fly is a pest of Asian origin that causes direct and indirect damages to citrus, damaging the development and production of plants. For the development of efficient management strategies of the pest, the integration of control methods is necessary, and biological control is the most appropriate. Among the agents that can be used, entomopathogenic fungi are considered one of the most important and wide-ranging use. This work investigated the occurrence of Purpureocillium lilacinum (Thom.) Luangsa-ard et al. (= Paecilomyces lilacinus), attackingnymphs of citrus black fly, Aleurocanthus woglumi Ashby (Hemiptera: Aleyrodidae). The fungus was isolated from infected Black fly nymphs, present on Citrus spp leaves in the municipality of Morros, Maranhão. After isolation, purification and morphological and molecular characterization, pathogenicity test was performed with A. woglumi nymphs. Morphological and molecular correspondence was verified between inoculum and the reisolated, proving the pathogenicity of P. lilacinum.

The rice rat Euryoryzomys russatus, a competent amplifying host of Rickettsia parkeri strain Atlantic rainforest for the tick Amblyomma ovale

Rickettsia parkeri strain Atlantic rainforest (SAR) is the etiological agent of a spotted fever group rickettsiosis in Brazil, where it is transmitted to humans by the tick Amblyomma ovale. A previous study demonstrated that R. parkeri SAR was successfully maintained in A. ovale ticks by transstadial and transovarial passages; however, because this agent induced lower reproduction rates in A. ovale, the participation of a vertebrate amplifier host, yet to be determined, was speculated. Since the rice rat Euryoryzomys russatus was demonstrated to be the most important host for immature stages of A. ovale in a focus of R. parkeri SAR transmission, the present study evaluated the competence of rice rats to act as amplifying host of R. parkeri SAR for A. ovale ticks. Rice rats were infested with R. parkeri SAR-infected A. ovale nymphs, and four days later with uninfected A. ovale larvae. Rickettsial transmission to rats was confirmed by seroconversion to R. parkeri antigens. Detached engorged larvae were allowed to molt to nymphs, in which rickettsial DNA was detected in up to 60% (mean: 20%) of the specimens. When part of these nymphs was allowed to feed on susceptible rice rats, rickettsial transmission was confirmed by seroconversion, indicating that there was successful horizontal transmission of R. parkeri SAR from infected nymphs to uninfected larvae in the previous acquisition infestations. Because we used naïve, susceptible rats, we infer that this horizontal transmission occurred via a systemic infection (rickettsemia) in the rat. Our results, coupled with previous epidemiological studies, suggest that under natural conditions rice rats could be acting as amplifying hosts of R. parkeri SAR to A. ovale ticks.

Effect of Climate and Land Use on the Spatio-Temporal Variability of Tick-Borne Bacteria in Europe.

The incidence of tick-borne diseases caused by Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Rickettsia spp. has been rising in Europe in recent decades. Early pre-assessment of acarological hazard still represents a complex challenge. The aim of this study was to model Ixodes ricinus questing nymph density and its infection rate with B. burgdorferi s.l., A. phagocytophilum and Rickettsia spp. in five European countries (Italy, Germany, Czech Republic, Slovakia, Hungary) in various land cover types differing in use and anthropisation (agricultural, urban and natural) with climatic and environmental factors (Normalized Difference Vegetation Index (NDVI), Normalized Difference Water Index (NDWI), Land Surface Temperature (LST) and precipitation). We show that the relative abundance of questing nymphs was significantly associated with climatic conditions, such as higher values of NDVI recorded in the sampling period, while no differences were observed among land use categories. However, the density of infected nymphs (DIN) also depended on the pathogen considered and land use. These results contribute to a better understanding of the variation in acarological hazard for Ixodes ricinus transmitted pathogens in Central Europe and provide the basis for more focused ecological studies aimed at assessing the effect of land use in different sites on tick–host pathogens interaction.

Experimental infection in Cavia porcellus by infected Amblyomma ovale nymphs with Rickettsia sp. (Atlantic rainforest strain).

This study describes experimental infection of guinea pigs (Cavia porcellus) infested with naturally infected Amblyomma ovale nymphs with Rickettsia sp. (Atlantic rainforest strain), and the capacity of A. ovale nymphs to transmit this bacterium. Twenty-six guinea pigs were divided into the following groups: G1, 10 animals infested with uninfected A. ovale nymphs; G2, 10 animals infested with nymphs infected with Rickettsia sp. (Atlantic rainforest strain); and G3, 6 animals without tick infestation. Blood samples were taken 7, 14, 21, and 28days post-infestation for serological and hematological tests. For histopathological analysis and rickettsial DNA detection, fragments of the spleen, lung, brain, and liver were harvested after euthanasia. The average feeding period for nymphs was 6.6days for G1 and 6days for G2. Hemolymph and PCR assays, performed to detect the causative agent in ticks, indicated that in G1, all ticks were negative, and in G2, all nymphs were positive by PCR and 80% (8/10) was positive by hemolymph tests. The only clinical change was skin scarring at the tick attachment site. Hematological parameters indicated leukopenia and total plasma protein (TPP) increased with decreased platelets in G1. In G2, leukocytosis, neutrophilia, monocytosis, an increase in platelets, and reduced TPP were observed. Only G2 guinea pigs were seroconverted (80%; 8/10). Histopathology tests indicated mild, diffuse hemosiderosis and mild, multifocal, follicular hyperplasia in the spleen. Molecular analysis did not detect Rickettsia sp. DNA in C. porcellus tissues. We demonstrated the capacity of A. ovale nymphs to transmit Rickettsia sp. (Atlantic rainforest strain) to guinea pigs.

Transstadial transmission and maintenance of Francisella tularensis in the Blacklegged Tick Ixodes scapularis.

In the south-central United States tularemia endemic region, approximately 10% of human cases occur during the fall-winter season when the questing activity of its primary tick vectors for this region Dermacentor variabilis and Amblyomma americanum is minimal. It is also been reported that approximately 40% of tularemia infected domestic cats contract the disease during the fall-winter season. To more fully characterize the potential role of I. scapularis, whose host seeking activity peaks during the fall-winter season in the south-central United States, as a vector for tularemia, the biology of F. tularensis subspecies holarctica strain LVS in colony-reared I. scapularis artificially infected with LVS by capillary feeding was studied. Capillary-fed I. scapularis larva and nymph were initially colonized with 1.9 × 103 and 1.1 × 104 CFU/tick, respectively. For nymphs molted from colonized larvae, the percent of molted nymphs colonized declined at the time of molting, but LVS persisted in 15% of molted nymphs up to 175 days post-capillary feeding. In contrast, only 5% of adults molted from infected nymphs maintained LVS colonization through 150 days post-capillary feeding. For capillary-fed adults, LVS initially colonized the gut and disseminated to hemolymph and salivary glands by 14 day post-feeding. Post-capillary feeding larvae and nymphs maintained LVS colonization initially and persisted although in a low percentage in molted nymphs and molted adults. Based on the feeding habits of I. scapularis adults, this tick species has the potential to act as a bridging vector for human tularemia for fall-winter cases of tularemia.

Vertical vs. horizontal transmission of the microbiome in a key disease vector, Ixodes pacificus

Vector-borne pathogens are increasingly found to interact with the vector's microbiome, influencing disease transmission dynamics. However, the processes that regulate the formation and development of the microbiome are largely unexplored for most tick species, an emerging group of disease vectors. It is not known how much of the tick microbiome is acquired through vertical transmission vs. horizontally from the environment or interactions with bloodmeal sources. Using 16S rRNA sequencing, we examined the microbiome of Ixodes pacificus, the vector of Lyme disease in the western USA, across life stages and infection status. We also characterized microbiome diversity in field and laboratory-collected nymphal ticks to determine how the surrounding environment affects microbiome diversity. We found a decrease in both species richness and evenness as the tick matures from larva to adult. When the dominant Rickettsial endosymbiont was computationally removed from the tick microbial community, we found that infected nymphs had lower species evenness than uninfected ticks, suggesting that lower microbiome diversity is associated with pathogen transmission in wild-type ticks. Furthermore, laboratory-reared nymph microbiome diversity was found to be compositionally distinct and significantly depauperate relative to field-collected nymphs. These results highlight unique patterns in the microbial community of I.pacificus that is distinct from other tick species. We provide strong evidence that ticks acquire a significant portion of their microbiome through exposure to their environment despite a loss of overall diversity through life stages. We provide evidence that loss of microbial diversity is at least in part due to elimination of microbial diversity with bloodmeal feeding but other factors may also play a role.

Infection of Ixodes ricinus by Borrelia burgdorferi sensu lato in peri-urban forests of France.

Lyme borreliosis is the most common tick-borne disease in the northern hemisphere. In Europe, it is transmitted by Ixodes ticks that carry bacteria belonging to the Borrelia burgdorferi sensu lato complex. The objective of this work was to explore eco-epidemiological factors of Lyme borreliosis in peri-urban forests of France (Sénart, Notre-Dame and Rambouillet). We investigated whether the introduction of Tamias sibiricus in Sénart could alter the density of infected ticks. Moreover, the density and tick infection were investigated according to the tree species found in various patches of Sénart forest. For this purpose, ticks were sampled during 3 years. In the Sénart forest, the density of nymph and adult ticks showed no significant difference between 2008, 2009 and 2011. The nymph density varied significantly as a function of the month of collection. Regarding the nymphs, a higher rate of infection and infected density were found in 2009. Plots with chipmunks (C) presented a lower density of both nymphs and adult ticks than plots without chipmunks (NC) did. A higher rate of infection of nymphs with Borrelia was seen in C plots. The prevalence of the various species of Borrelia was also found to vary between C and NC plots with the year of the collect. The presence of chestnut trees positively influenced the density of both nymphs and adults. The infected nymph density showed a significant difference depending on the peri-urban forest studied, Sénart being higher than Rambouillet. The prevalence of Borrelia species also differed between the various forests studied. Concerning the putative role that Tamias sibiricus may play in the transmission of Borrelia, our results suggest that its presence is correlated with a higher rate of infection of questing ticks by Borrelia genospecies and if its population increases, it could play a significant role in the risk of transmission of Lyme borreliosis.

Year-to-year variation in the density of Ixodes ricinus ticks and the prevalence of the rodent-associated human pathogens Borrelia afzelii and B. miyamotoi in different forest types

The human pathogens Borrelia afzelii, which causes Lyme borreliosis and B. miyamotoi, which causes relapsing fever, both circulate between Ixodes ricinus ticks and rodents. The spatiotemporal dynamics in the prevalence of these pathogens have not yet been fully elucidated, but probably depend on the spatiotemporal population dynamics of small rodents. We aimed to evaluate the effect of different forest types on the density of infected nymphs in different years and to obtain more knowledge about the spatial and temporal patterns of ticks and tick-borne pathogens. We analysed unfed nymphal ticks from 22 stands of four different forest types in Belgium in 2009, 2010, 2013 and 2014 and found that the density of nymphs in general and the density of nymphs infected with B. afzelii and B. miyamotoi varied yearly, but without temporal variation in the infection prevalence. The yearly variation in density of infected nymphs in our study thus seems to be caused most by the variation in the density of nymphs, which makes it a good predictor of disease risk. The risk for rodent-associated tick-borne diseases also varied between forest types. We stress the need to elucidate the contribution of the host community composition to tick-borne disease risk.

Cascading effects of predator activity on tick-borne disease risk.

Predators and competitors of vertebrates can in theory reduce the density of infected nymphs (DIN)—an often-used measure of tick-borne disease risk—by lowering the density of reservoir-competent hosts and/or the tick burden on reservoir-competent hosts. We investigated this possible indirect effect of predators by comparing data from 20 forest plots across the Netherlands that varied in predator abundance. In each plot, we measured the density of questing Ixodes ricinus nymphs (DON), DIN for three pathogens, rodent density, the tick burden on rodents and the activity of mammalian predators. We analysed whether rodent density and tick burden on rodents were correlated with predator activity, and how rodent density and tick burden predicted DON and DIN for the three pathogens. We found that larval burden on two rodent species decreased with activity of two predator species, while DON and DIN for all three pathogens increased with larval burden on rodents, as predicted. Path analyses supported an indirect negative correlation of activity of both predator species with DON and DIN. Our results suggest that predators can indeed lower the number of ticks feeding on reservoir-competent hosts, which implies that changes in predator abundance may have cascading effects on tick-borne disease risk.