Can the Gram-negative bacterium Escherichia coli colonize the gut of Lone Star Tick (Amblyomma americanum)?

Ticks are obligate blood feeding ectoparasites and vectors of several mammalian pathogens (Williams-Newkirk et al, 2014). In addition to pathogens they also carry a bacterial community with commensal and symbiotic relationships (Bonnet et al, 2017). Using a culture-dependent approach we previously reported a high prevalence of Gram-positive bacteria in the gut of field collected lone star ticks (Amblyomma americanum). These results suggested that epithelial immunity functions to control Gram-negative bacteria in A. americanum. In this study, we used a culturing and non-culturing approach to measure the outcome of E.coli (Gram-negative) when fed to female adult lone star ticks (n=16). Results showed a significant reduction of E.coli at Days 1, 3 and 7 post bacterial feeding. qPCR of 16S rDNA confirmed reduction of bacterial rDNA when compared to water fed ticks (n=16). Our results suggest that there is a midgut epithelial immune response in place, which mainly targets Gram-negative bacteria

Geospatial Analysis of Rickettsial Species

Rickettsia species are obligate intracellular, arthropod-borne bacteria with a potential to cause multiple diseases including Rocky Mountain spotted fever (RMSF). Fleas, mites, and ticks serve as vectors for Rickettsia, but ticks are the primary vector of interest. RMSF and other rickettsial diseases have continued to gain importance in both human and veterinary medicine as RMSF is the most common tick-borne disease within the United States according to the Lyme and Tick-Borne Disease Research Center. A statewide citizen science project was utilized to determine the prevalence of Spotted Fever Group (SFG) Rickettsia in Arkansas. This project yielded results in 64 of Arkansas’s 75 counties. Results were utilized to determine prevalence in each of the represented counties, and then compiled into a geospatial representation of the data. It was determined that 34.32% of the ticks sampled were carriers of one or more rickettsial species. As the samples were divided by county, multiple counties were shown to have concerningly high exposure risk for SFG Rickettsia. There were six species of ticks represented throughout this study with Amblyomma americanum being the most common. There were also six species of Spotted Fever Group Rickettsia found within the samples. The small portion of ticks that underwent further analysis to determine the specific rickettsial species present, indicated that Rickettsia amblyommatis is likely the most common SFG Rickettsia in Arkansas

Metagenomic-based Surveillance of Pacific Coast tick Dermacentor occidentalis Identifies Two Novel Bunyaviruses and an Emerging Human Ricksettsial Pathogen.

An increasing number of emerging tick-borne diseases has been reported in the United States since the 1970s. Using metagenomic next generation sequencing, we detected nucleic acid sequences from 2 novel viruses in the family Bunyaviridae and an emerging human rickettsial pathogen, Rickettsia philipii, in a population of the Pacific Coast tick, Dermacentor occidentalis in Mendocino County sampled annually from 2011 to 2014. A total of 250 adults of this human-biting, generalist tick were collected from contiguous chaparral and grassland habitats, and RNA from each individually extracted tick was deep sequenced to an average depth of 7.3 million reads. We detected a Francisella endosymbiont in 174 ticks (70%), and Rickettsia spp. in 19 ticks (8%); Rickettsia-infected ticks contained R. rhipicephali (16 of 250, 6.4%) or R. philipii (3 of 250,1.2%), the agent of eschar-associated febrile illness in humans. The genomes of 2 novel bunyaviruses (>99% complete) in the genera Nairovirus and Phlebovirus were also identified and found to be present in 20-91% of ticks, depending on the year of collection. The high prevalence of these bunyaviruses in sampled Dermacentor ticks suggests that they may be viral endosymbionts, although further studies are needed to determine whether they are infectious for vertebrate hosts, especially humans, and their potential role in tick ecology

High prevalence of <i>Rickettsia africae</i> variants in <i>Amblyomma variegatum</i> ticks from domestic mammals in rural western Kenya: implications for human health

Tick-borne spotted fever group (SFG) rickettsioses are emerging human diseases caused by obligate intracellular Gram-negative bacteria of the genus Rickettsia. Despite being important causes of systemic febrile illnesses in travelers returning from sub-Saharan Africa, little is known about the reservoir hosts of these pathogens. We conducted surveys for rickettsiae in domestic animals and ticks in a rural setting in western Kenya. Of the 100 serum specimens tested from each species of domestic ruminant 43% of goats, 23% of sheep, and 1% of cattle had immunoglobulin G (IgG) antibodies to the SFG rickettsiae. None of these sera were positive for IgG against typhus group rickettsiae. We detected Rickettsia africae–genotype DNA in 92.6% of adult Amblyomma variegatum ticks collected from domestic ruminants, but found no evidence of the pathogen in blood specimens from cattle, goats, or sheep. Sequencing of a subset of 21 rickettsia-positive ticks revealed R. africae variants in 95.2% (20/21) of ticks tested. Our findings show a high prevalence of R. africae variants in A. variegatum ticks in western Kenya, which may represent a low disease risk for humans. This may provide a possible explanation for the lack of African tick-bite fever cases among febrile patients in Kenya

Neuronal in vitro impact of Amblyommaamericanumsalivary glands extracts

When a tick feeds off a host, the salivary glands of the tick excrete saliva to assist the tick in feeding (1). The reason saliva assists the tick in consuming its blood meal is due to its immunosuppressive, anti-inflammatory, and anti-coagulant properties (2).Tick salivary glands also play an important role in the transmission of tick-borne pathogens (3). We wanted to investigate the effect of tick salivary gland extracts (SGE) on human neurons. For our experiment, the organism whose salivary gland extract we used was the Amblyomma americanum. We compared the effects of salivary gland extract from ticks collected in the field and ticks reared in a lab colony. Our results showed that the salivary gland extract from the two types of ticks affected the neurons differently, and the salivary gland extract of the field tick was more damaging to the neurons. This indicates that SGE from field ticks, together with infectious agents, could result in a more severe neuronal impairment in a human host than the infectious agent alone

Tick Sweats

Ticks are obligatory ectoparasites of many vertebrate hosts including human. Osmoregulatory functions of ticks are crucial for the survival, especially, in the off-host ticks in arid area. We found that injection of water in the body cavity of tick immediately triggers excretion of solution through the exoskeletal cuticles, like sweating. This response occurred in a bilateral asymmetric manner; the injection on left side of the body induced the sweating on only the left half, while the injection into right side did not induce sweat. The sweating response was reduced in the injections of high osmolar Naci (1 M). This is the first description of sweating physiology in maintenance of water homeostasis in the Lone star tick

Identification of a mutation in the para-sodium channel gene of the cattle tick Rhipicephalus (Boophilus) microplus associated with resistance to synthetic pyrethroid acaricides

Resistance against synthetic pyrethroid (SP) products for the control of cattle ticks in Australia was detected in the field in 1984, within a very short time of commercial introduction. We have identified a mutation in the domain II S4-5 linker of the para-sodium channel that is associated with resistance to SPs in the cattle tick Rhipicephalus (Boophilus) microplus from Australia. The cytosine to adenine mutation at position 190 in the R. microplus sequence AF134216, results in an amino acid substitution from leucine in the susceptible strain to isoleucine in the resistant strain. A similar mutation has been shown to confer SP resistance in the whitefly, Bemisia tabaci, but has not been described previously in ticks. A diagnostic quantitative PCR assay has been developed using allele-specific Taqman® minor groove-binding (MGB) probes. Using the assay to screen field and laboratory populations of ticks showed that homozygote allelic frequencies correlated highly with the survival percentage at the discriminating concentration of cypermethrin

Geospatial Analysis of Rickettsial Species in Arkansas

Rickettsia species are obligate intracellular, arthropod-borne bacteria with a potential to cause multiple diseases including Rocky Mountain spotted fever (RMSF). Fleas, mites, and ticks serve as vectors for Rickettsia, but ticks are the primary vector of interest. RMSF and other rickettsial diseases have continued to gain importance in both human and veterinary medicine as RMSF is the most common tick-borne disease within the United States according to the Lyme and Tick-Borne Disease Research Center. A statewide citizen science project was utilized to determine the prevalence of Spotted Fever Group (SFG) Rickettsia in Arkansas. This project yielded results in 64 of Arkansas’s 75 counties. Results were utilized to determine prevalence in each of the represented counties, and then compiled into a geospatial representation of the data. It was determined that 34.32% of the ticks sampled were carriers of one or more rickettsial species. As the samples were divided by county, multiple counties were shown to have concerningly high exposure risk for SFG Rickettsia. There were six species of ticks represented throughout this study with Amblyomma americanum being the most common. There were also six species of Spotted Fever Group Rickettsia found within the samples. The small portion of ticks that underwent further analysis to determine the specific rickettsial species present, indicated that Rickettsia amblyommatis is likely the most common SFG Rickettsia in Arkansas

Survival of Theileria parva in its nymphal tick vector Rhipicephalus appendiculatus under laboratory and quasi natural conditions

Groups of nymphal Rhipicephalus appendiculatus Muguga, having a mean of 1 or 9 Theileria parva Muguga-infected salivary gland acini per tick, were kept under quasi-natural conditions at an altitude of 1950 m or 20°C at a relative humidity of 85% in the laboratory and their survival and infection prevalence and abundance determined over time. Theileria parva infections for both categories of ticks survived in the nymphal ticks for 50 or 26 weeks post salivary gland infection under quasi-natural or laboratory conditions respectively. There was a distinct decline in infections in the more heavily infected nymphae under both conditions of exposure, reflecting an apparent density dependence in parasite survival. Nymphal ticks having an average infection level of 1 infected salivary gland acinus per tick, survived for up to 69 or 65 weeks post-repletion under quasi-natural or the laboratory conditions respectively. Nymphae having an average infection level of 9 infected salivary gland acini per tick survived for a similar duration under each of the 2 conditions. The infection level of 9 infected salivary gland acini per tick did not seem to significantly affect the survival of the tick vector compared to those having an average of 1 infected salivary gland acinus per tick

Bartonella infections in fleas (Siphonaptera : Pulicidae) and lack of Bartonellae in ticks (Acari : Ixodidae) from Hungary

Fleas (95 Pulex irritans, 50 Ctenocephalides felis, 45 Ctenocephalides canis) and ixodid ticks (223 Ixodes ricinus, 231 Dermacentor reticulatus, 204 Haemaphysalis concinna) were collected in Hungary and tested, in assays based on PCR, for Bartonella infection. Low percentages of P. irritans (4.2%) and C. felis (4.0%) were found to be infected. The groEL sequences of the four isolates from P. irritans were different from all the homologous sequences for bartonellae previously stored in GenBank but closest to those of Bartonella sp. SE-Bart-B (sharing 96% identities). The groEL sequences of the two isolates from C. felis were identical with those of the causative agents of cat scratch disease, Bartonella henselae and Bartonella clarridgeiae, respectively. The pap31 sequences of B. henselae amplified from Hungarian fleas were identical with that of Marseille strain. No Bartonella-specific amplification products were detected in C. canis, L ricinus, D. reticulatus and H. concinna pools