Amblyomma maculatum-associated rickettsiae in vector tissues and vertebrate hosts during tick feeding.

Rickettsia parkeri, a causative agent of spotted fever rickettsiosis, is transmitted by Amblyomma maculatum (Gulf Coast tick), a tick that may also carry a non-pathogenic spotted fever group Rickettsia, "Candidatus Rickettsia andeanae". Here, we evaluated R. parkeri and "Candidatus R. andeanae" in tissues from A. maculatum prior to, during, and after blood feeding on rabbits. Using colony-reared A. maculatum that were capillary-fed uninfected cells, R. parkeri, "Candidatus R. andeanae", or both rickettsiae, we detected higher levels of Rickettsia spp. in the respective treatment groups. Rickettsial levels increased during blood feeding for both R. parkeri and "Candidatus R. andeanae", with a greater increase in R. parkeri in co-infected ticks compared to singly-infected ticks. We detected transovarial transmission of "Candidatus R. andeanae" in egg and larval cohorts and confirmed vertical transmission of R. parkeri in one group of larvae. Rabbits from all Rickettsia-exposed groups seroconverted on immunofluorescent antibody testing using R. parkeri antigen. Visualization of "Candidatus R. andeanae" in tick salivary glands suggested potential transmission via tick feeding. Here, rickettsial levels in artificially infected ticks demonstrate changes during feeding and transovarial transmission that may be relevant for interpreting rickettsial levels detected in wild A. maculatum.

A Statewide Survey for Container-Breeding Mosquitoes in Mississippi.

Container-breeding mosquitoes are important in public health due to outbreaks of Zika, chikungunya, and dengue viruses. This paper documents the distribution of container-breeding mosquito species in Mississippi, with special emphasis on the genus Aedes. Five sites in each of the 82 Mississippi counties were sampled monthly between May 1 and August 31, 2016, and 50,109 mosquitoes in 14 species were collected. The most prevalent and widely distributed species found was Ae. albopictus, being found in all 82 counties, especially during July. A recent invasive, Ae. japonicus, seems to be spreading rapidly in Mississippi since first being discovered in the state in 2011. The most abundant Culex species collected were Cx. quinquefasciatus (found statewide), Cx. salinarius (almost exclusively in the southern portion of the state), and Cx. restuans (mostly central and southern Mississippi). Another relatively recent invasive species, Cx. coronator, was found in 20 counties, predominantly in the southern one-third of the state during late summer. Co-occurrence data of mosquito species found in the artificial containers were also documented and analyzed. Lastly, even though we sampled extensively in 410 sites across Mississippi, no larval Ae. aegypti were found. These data represent the first modern statewide survey of container species in Mississippi, and as such, allows for better public health readiness for emerging diseases and design of more effective vector control programs.

Seasonality of Aedes albopictus in North and Central Mississippi.

Container-breeding mosquitoes are increasingly important in public health due to recent outbreaks of Zika virus, chikungunya, and dengue. This paper documents seasonality of the most prevalent container-breeding mosquito species in Mississippi-Aedes albopictus. Ten sites in 5 counties in both northern and central Mississippi (20 sites, 10 counties total) were sampled by larval dipping and oviposition traps biweekly from September 2016 to June 2019, totaling 22 months and potentially yielding 440 egg or larval collections. However, 22 collections were missed due to inclement weather and personnel issues during the study period, so actually only 418 site visits were performed. Sites were chosen to maximize chances of finding Ae. albopictus. Of the total 1,310 mosquito larvae collected during the study period, 717 larvae and 50 positive egg papers belonged to Ae. albopictus. Aedes albopictus was found in all 10 northern and central counties. No eggs were collected at any of the sites from December through February, although larvae were occasionally collected during that time frame. This study demonstrates that Ae. albopictus is active in central and northern Mississippi beginning in March each year and continuing through November or December. There is little activity during the coldest months of the year (January and February). These data represent the first extensive analysis of Ae. albopictus seasonality in Mississippi, and as such, allow for better public health awareness of diseases transmitted by this species and design of more effective vector control programs.

What Is Going on With the Genus Dermacentor? Hybridizations, Introgressions, Oh My!

In the 1930s, R. A. Cooley noted that Dermacentor occidentalis (Acarina: Ixodidae) and Dermacentor andersoni were closely related and could hybridize. Decades later, James Oliver discovered that crosses of Dermacentor variabilis, D. andersoni, and D. occidentalis could, on occasion, produce hybrids. A recent molecular analysis (both mtDNA and nDNA) in our laboratory revealed that certain specimens of Dermacentor andersoni nested with Dermacentor parumapertus. Does this close relationship, along with the mito-nuclear discordance we have observed, mean D. andersoni and D. parumapertus are a single species? By contemporary taxonomic criteria, this seems improbable based on their distinctly different morphologies, host associations, and ecologies. This paper explores ideas related to mito-nuclear discordance, hybridization, and introgression (primarily) not only in these two species but also other members of the genus Dermacentor. Both D. andersoni and D. parumapertus can be found on the same hosts and have sympatric distributions, so introgression of genetic material by occasional cross-mating between these two species is possible. Further, the difficulty in applying specific species concepts in ticks has been recently pointed out and a unified agreement on an integrative species concepts could clearly be useful in this situation. With the discovery of D. parumapertus as a potential vector of Rickettsia parkeri and the historically recognized role of D. andersoni in transmission of Rickettsia rickettsii, understanding the specific status of each lineage of these species (and others in the genus) is extremely important from a public health perspective.

Molecular Phylogeny of Dermacentor parumapertus (Acari: Ixodidae) from Two Locations Within Its Geographical Range.

Dermacentor parumapertus Neumann (Acari: Ixodidae) is a rather obscure tick found on jackrabbits in the western United States and parts of Canada and Mexico. Specimens from the northern part of their range are consistently different morphologically from ones found in southern and eastern parts of their range (particularly west Texas), leading some researchers to declare the southern form a variety or subspecies. This study examined field-collected adult D. parumapertus from two main locations-Utah and Texas-within its geographic distribution to ascertain the degree of genetic divergence in the two populations based upon both nuclear and mitochondrial DNA sequences. In total, 30 D. parumapertus were analyzed by PCR using both mtDNA and nDNA genes, and one D. nitens was included for comparison. Trees were constructed for all mtDNA genes individually, as well as after concatenating mtDNA (COI, COII, 12S) and nDNA (2 ITS2 primers), respectively. All constructed trees were exported to FigTree v1.4.3 and TreeGraph v2.14.1-771 beta for visualization. The majority of the Utah and Texas populations of D. parumapertus separated molecularly in both mtDNA and nDNA trees; however, analysis with mtDNA genes showed that 3/13 (23%) of Utah tick specimens were removed molecularly from other specimens collected at the same location. Thus, there was not enough evidence to declare these two disparate and morphologically different populations as distinct and separate species.

Morphometric Analysis of Adult Dermacentor parumapertus Neumann (Acari: Ixodidae) From Various Locations Within its Geographical Range.

Dermacentor parumapertus Neumann (Acari: Ixodidae), a tick primarily associated with rabbits which occurs over much of the western United States, has a fairly large north-to-south distribution, being found from central Idaho southward into northern Mexico. This mostly obscure tick species has recently been the focus of attention due to the discovery of a unique strain of Rickettsia parkeri associated with it which appears closely related to a Rickettsia sp. found in the Atlantic rainforest of Brazil. Historically, a morphological variety of this species was reported in the literature based on significant variation in ornamentation of the tick throughout its range. This study examines several key morphological characters to determine if there are indeed more than one distinct population of this species throughout its range.

Transmission of Amblyomma maculatum-Associated Rickettsia spp. During Cofeeding on Cattle.

Amblyomma maculatum is the primary vector for the spotted fever group rickettsiae, Rickettsia parkeri, a known pathogen, and "Candidatus Rickettsia andeanae," currently considered nonpathogenic. Spotted fever group rickettsiae are typically endothelial cell associated and rarely circulate in the blood. Horizontal transmission to naïve ticks through blood feeding from an infected host is likely rare. Cofeeding provides an opportunity for rickettsial transmission to naïve ticks in the absence of circulating rickettsiae. We evaluated R. parkeri transmission through cofeeding between A. maculatum adults and nymphs on beef calves. Six calves in each of two trials were infested with A. maculatum that had been capillary fed R. parkeri. Four days later, calves each received recipient A. maculatum that were either capillary fed "Ca. R. andeanae" or not capillary fed before infestation. Trials differed by whether we included a barrier to minimize adjacent feeding between recipient and donor ticks. After cofeeding, we detected R. parkeri in 27% of "Ca. R. andeanae"-free recipient ticks, whereas R. parkeri was not detected in any recipient ticks that were capillary fed "Ca. R. andeanae." Rickettsia parkeri transmission efficiency to naïve ticks was greater when ticks freely cofed in proximity. No rickettsial DNA was detected in calf blood. Results confirm cofeeding as a method of horizontal transmission of R. parkeri in the absence of host rickettsemia and suggest no evidence of transmission by cofeeding when recipient ticks are first exposed to "Ca. R. andeanae" through capillary feeding. While cofeeding may provide an opportunity for maintaining the pathogen, R. parkeri, the mechanisms driving any potential effect of "Ca. R. andeanae" on R. parkeri transmission are unclear.

Differentiated THP-1 Cells Exposed to Pathogenic and Nonpathogenic Borrelia Species Demonstrate Minimal Differences in Production of Four Inflammatory Cytokines.

Tick-borne borreliae include Lyme disease and relapsing fever agents, and they are transmitted primarily by ixodid (hard) and argasid (soft) tick vectors, respectively. Tick-host interactions during feeding are complex, with host immune responses influenced by biological differences in tick feeding and individual differences within and between host species. One of the first encounters for spirochetes entering vertebrate host skin is with local antigen-presenting cells, regardless of whether the tick-associated Borrelia sp. is pathogenic. In this study, we performed a basic comparison of cytokine responses in THP-1-derived macrophages after exposure to selected borreliae, including a nonpathogen. By using THP-1 cells, differentiated to macrophages, we eliminated variations in host response and reduced the system to an in vitro model to evaluate the extent to which the Borrelia spp. influence cytokine production. Differentiated THP-1 cells were exposed to four Borrelia spp., Borrelia hermsii (DAH), Borrelia burgdorferi (B31), B. burgdorferi (NC-2), or Borrelia lonestari (LS-1), or lipopolysaccharides (LPS) (activated) or media (no treatment) controls. Intracellular and secreted interferon (IFN)-γ, interleukin (IL)-1ß, IL-6, and tumor necrosis factor (TNF)-α were measured using flow cytometric and Luminex-based assays, respectively, at 6, 24, and 48 h postexposure time points. Using a general linear model ANOVA for each cytokine, treatment (all Borrelia spp. and LPS compared to no treatment) had a significant effect on secreted TNF-α only. Time point had a significant effect on intracellular IFN-γ, TNF-α and IL-6. However, we did not see significant differences in selected cytokines among Borrelia spp. TREATMENTS: Thus, in this model, we were unable to distinguish pathogenic from nonpathogenic borreliae using the limited array of selected cytokines. While unique immune profiles may be detectable in an in vitro model and may reveal predictors for pathogenicity in borreliae of unknown pathogenicity, a larger panel of cytokines would be desirable to test.