Ecological release and patch geometry can cause nonlinear density-area relationships.

A primary driver of species extinctions and declining biodiversity is loss and fragmentation of habitats owing to human activities. Many studies spanning a wide diversity of taxa have described the relationship between population density and habitat patch area, i.e., the density-area relationship (DAR), as positive, neutral, negative or some combination of the three. However, the mechanisms responsible for these relationships remain elusive. We employ a theoretical spatially explicit population model built upon the reaction-diffusion framework with absorbing boundary conditions to model a habitat specialist dwelling in islands of habitat surrounded by a hostile matrix. We consider patches with a convex or non-convex geometry. Our results show that a single species following logistic-type population growth exhibits a strictly positive and continuous DAR. However, when multiple asymptotically stable steady states are preset in the system, a discontinuous DAR arises. In the case of two species governed by diffusive Lotka-Volterra growth and competitive interactions, we observe that overall DAR structure can be either (1) positive, (2) positive for small areas and neutral for large, or (3) hump-shaped, i.e., positive for area below a threshold and negative for area above. Patch complexity such as non-convex geometry can cause discontinuities in DAR slope for a single species and create qualitatively different patterns in a competitive system as compared to a convex patch. We also compared our theoretical results with two empirical studies (Anolis lizards on islands and crossbills and pine squirrels in forest fragments) where the pragmatic view of DAR fails to give a mechanistic understanding of what was observed. Close qualitative agreement between theoretical and observed DAR indicates that our model gives a reasonable explanation of the mechanisms underpinning DAR found in those studies. From a conservation perspective, the DAR is crucial to the identification of valuable habitat fragments that favor high abundance and the design of a reserve for a target species. When it comes to protecting a single species, these results suggest that there is unlikely to be a simple solution and that conservation decisions should always be made on a case-by-case basis.

Frequency of Occurrence and Population-Dynamic Consequences of Different Forms of Density-Dependent Emigration.

Emigration is a fundamental process affecting species' local, regional, and large-scale dynamics. The paradigmatic view in ecology is that emigration is density independent (DIE) or positive density dependent (+DDE). However, alternative forms are biologically plausible, including negative (-DDE), U-shaped (uDDE), and hump-shaped (hDDE) forms. We reviewed the empirical literature to assess the frequency of different forms of density-dependent emigration and whether the form depended on methodology. We also developed a reaction-diffusion model to illustrate how different forms of DDE can affect patch-level population persistence. We found 145 studies, the majority representing DIE (30%) and +DDE (36%). However, we also regularly found -DDE (25%) and evidence for nonlinear DDE (9%), including one case of uDDE and two cases of hDDE. Nonlinear DDE detection is likely hindered by the use of few density levels and small density ranges. Based on our models, DIE and +DDE promoted stable and persistent populations. uDDE and -DDE generated an Allee effect that decreases minimum patch size. Last, -DDE and hDDE models yielded bistability that allows the establishment of populations at lower densities. We conclude that the emigration process can be a diverse function of density in nature and that alternative DDE forms can have important consequences for population dynamics.

Human immunoglobulin G responses to Cimex lectularius L. saliva.

AIMS: To investigate the immunoglobulin (Ig) G response after being fed upon by Cimex lectularius L. METHODS AND RESULTS: Participants were fed upon by three male C lectularius insects weekly for a month. Blood was obtained before the feeding and at the last feeding, which was used for immunoblots against bed bug salivary gland extract, with antihuman Immunoglobulin G (IgG) secondary antibodies. No consistent IgG changes developed in 11 humans serially fed upon by C lectularius. Two participants had new IgG responses to proteins at molecular weights of approximately 12-13 kDa, and one had an IgG response to a protein at approximately 40 kDa. At the last study visit, more intense IgG bands to proteins at molecular weights of 12-13 kDa had developed in 55% of participants (6/11) and at molecular weights of ≈30, ≈40 and ≈70 kDa in 45% (5/11) compared with the first study visit. Nitrophorin and apyrase were the most common C lectularius proteins identified with liquid chromatography-tandem mass spectrometry in both crushed bed bug salivary gland extract and post-bed bug feeding extract. CONCLUSIONS: Human participants did not have consistent IgG responses to crushed C lectularius salivary gland extract.

Reproductive incompatibility between Amblyomma maculatum (Acari: Ixodidae) group ticks from two disjunct geographical regions within the USA.

The Amblyomma maculatum Koch group of ixodid ticks consists of three species: A. maculatum, A. triste, and A. tigrinum. However, since Koch described this group in 1844, the systematics of its members has been the subject of ongoing debate. This is especially true of A. maculatum and A. triste; recent molecular analyses reveal insufficient genetic divergence to separate these as distinct species. Further confounding this issue is the discovery in 2014 of A. maculatum group ticks in southern Arizona (AZ), USA, that share morphological characteristics with both A. triste and A. maculatum. To biologically evaluate the identity of A. maculatum group ticks from southern Arizona, we analyzed the reproductive compatibility between specimens of A. maculatum group ticks collected from Georgia (GA), USA, and southern Arizona. Female ticks from both Arizona and Georgia were mated with males from both the Georgia and Arizona Amblyomma populations, creating two homologous and two heterologous F1 cohorts of ticks: GA ♀/GA ♂, AZ ♀/AZ ♂, GA ♀/AZ ♂, and AZ ♀/GA ♂. Each cohort was maintained separately into the F2 generation with F1 females mating only with F1 males from their same cohort. Survival and fecundity parameters were measured for all developmental stages. The observed survival parameters for heterologous cohorts were comparable to those of the homologous cohorts through the F1 generation. However, the F1 heterologous females produced F2 egg clutches that did not hatch, thus indicating that the Arizona and Georgia populations of A. maculatum group ticks tested here represent different biological species.

Effects of Patch-Matrix Composition and Individual Movement Response on Population Persistence at the Patch Level.

Fragmentation creates landscape-level spatial heterogeneity which in turn influences population dynamics of the resident species. This often leads to declines in abundance of the species due to increased susceptibility to edge effects between the remnant habitat patches and the lower quality "matrix" surrounding these focal patches. In this paper, we formalize a framework to facilitate the connection between small-scale movement and patch-level predictions of persistence through a mechanistic model based on reaction-diffusion equations. The model is capable of incorporating essential information about edge-mediated effects such as patch preference, movement behavior, and matrix-induced mortality. We mathematically analyze the model's predictions of persistence with a general logistic-type growth term and explore their sensitivity to demographic attributes in both the patch and matrix, as well as patch size and geometry. Also, we provide bounds on demographic attributes and patch size in order for the model to predict persistence of a species in a given patch based on assumptions on the patch/matrix interface. Finally, we illustrate the utility of this framework with a well-studied planthopper species (Prokelisia crocea) living in a highly fragmented landscape. Using experimentally derived data from various sources to parameterize the model, we show that, qualitatively, the model results are in accord with experimental predictions regarding minimum patch size of P. crocea. Through application of a sensitivity analysis to the model, we also suggest a ranking of the most important model parameters based on which parameter will cause the largest output variance.

Unique Strain of Rickettsia parkeri Associated with the Hard Tick Dermacentor parumapertus Neumann in the Western United States.

In 1953, investigators at the Rocky Mountain Laboratories in Hamilton, MT, described the isolation of a spotted fever group Rickettsia (SFGR) species from Dermacentor parumapertus ticks collected from black-tailed jackrabbits (Lepus californicus) in northern Nevada. Several decades later, investigators characterized this SFGR (designated the parumapertus agent) by using mouse serotyping methods and determined that it represented a distinct rickettsial serotype closely related to Rickettsia parkeri; nonetheless, the parumapertus agent was not further characterized or studied. To our knowledge, no isolates of the parumapertus agent remain in any rickettsial culture collection, which precludes contemporary phylogenetic placement of this enigmatic SFGR. To rediscover the parumapertus agent, adult-stage D. parumapertus ticks were collected from black-tailed jackrabbits shot or encountered as roadkills in Arizona, Utah, or Texas from 2011 to 2016. A total of 339 ticks were collected and evaluated for infection with Rickettsia species. Of 112 D. parumapertus ticks collected in south Texas, 16 (14.3%) contained partial ompA sequences with the closest identity (99.6%) to Rickettsia sp. strain Atlantic rainforest Aa46, an SFGR that is closely related or identical to an SFGR species that causes a mild rickettsiosis in several states of Brazil. A pure isolate, designated strain Black Gap, was cultivated in Vero E6 cells, and sequence analysis of the rrs, gltA, sca0, sca5, and sca4 genes also revealed the closest genetic identity to Rickettsia sp. Atlantic rainforest Aa46. Phylogenetic analysis of the five concatenated rickettsial genes place Rickettsia sp. strain Black Gap and Rickettsia sp. Atlantic rainforest Aa46 with R. parkeri in a distinct and well-supported clade.IMPORTANCE We suggest that Rickettsia sp. Black Gap and Rickettsia sp. Atlantic rainforest Aa46 represent nearly identical strains of R. parkeri and that Rickettsia sp. Black Gap or a very similar strain of R. parkeri represents the parumapertus agent. The close genetic relatedness among these taxa, as well as the response of guinea pigs infected with the Black Gap strain, suggests that R. parkeri Black Gap could cause disease in humans. The identification of this organism could also account, at least in part, for the remarkable differences in severity ascribed to Rocky Mountain spotted fever (RMSF) among various regions of the American West during the early 20th century. We suggest that the wide variation in case fatality rates attributed to RMSF could have occurred by the inadvertent inclusion of cases of milder disease caused by R. parkeri Black Gap.

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.

Evaluation of Human Attachment by Larval Amblyomma maculatum (Acari: Ixodidae).

The tick, Amblyomma maculatum Koch (Gulf Coast tick), has recently been shown to be an important disease vector of both medical and veterinary concern. Although much is known about the behavior and ecology of adults, little is known of the immatures. Larval feeding on humans has never been demonstrated (and thus, there are no collection records from humans). In this experiment, 10 larval A. maculatum, Amblyomma americanum (L.) (a positive control), and Dermacentor variabilis (Say) (a negative control), were applied to both forearms of 10 human volunteers (five male, five female). Ticks were placed in plastic caps and secured to skin with medical-grade adhesive tape, and volunteers remained sedentary during the experiment. After 15 min, caps were removed, and attachment was determined using fine-tipped forceps. Any A. maculatum that were attached were then removed and subsequently examined microscopically to verify identification. In total, 34 ticks attached to the subjects, including 11 A. maculatum (5.5%), 23 A. americanum (11.5%), and no D. variabilis. Amblyomma maculatum attached to six volunteers, and no apparent association between gender and attachment rate was noted. No skin lesions developed in the human volunteers bit by A. maculatum. This is the first report of larval A. maculatum attaching to humans, and is significant in that Rickettsia parkeri, a human pathogen vectored by this species, has recently been reported to be transmitted transovarially. If A. maculatum are infected as larvae, they could potentially transmit R. parkeri to people.

Use of the VectorTest for Advanced Warning of Human West Nile Virus Cases in Mississippi.

West Nile virus (WNV) continues to persist in Mississippi; 2012 was the worse year for human infections, with a total of 247 reported human cases and five deaths. Public health officials are keenly interested in ways to detect WNV in advance in their jurisdictions, so they can implement appropriate and timely mosquito control in affected areas. A total of 40,312 female Culex quinquefasciatus mosquitoes were collected by gravid traps in Mississippi in 2013 and 2014 and tested by VectorTest, a rapid immunochromatographic assay ("dip-stick" test) that is a highly specific and effective rapid threat assessment tool. This study evaluated if and to what extent VectorTest could provide advanced warning of impending human WNV cases in a specific area. These data were examined with regard to date of onset of human WNV cases to determine the predictive value of VectorTest for WNV activity. Both years, positive mosquito pools appeared before the vast majority (87.2%) of reported human cases. Overall, in 27 out of 37 human WNV cases (73.0 %) occurring in our study sites, there was an average advanced warning of 26 days (range 11­53 days) as indicated by positive mosquito collections near the patient's home. This operational health department study, although somewhat limited, reveals that mosquito sampling and testing can inform public health and mosquito control personnel of WNV activity in an area and of impending human cases.

Chikungunya in Mississippi: The Health Department Response to Imported Cases.

Chikungunya (CHIK), a newly recognized mosquito-borne disease in the Western Hemisphere, has resulted in well over a million cases since December 2013. Only about a dozen locally-acquired cases thus far have been reported in the U. S. (Florida), but approximately 1500 imported cases have been seen in returning travelers from the Caribbean and Central and South America. Public health officials are concerned that imported cases may lead to infection of local mosquitoes and, thus disease transmission. This paper documents 9 confirmed CHIK cases in Mississippi: 5 resulting from travel to the Dominican Republic, 2 from Haiti, 1 from Honduras, and 1 from Puerto Rico. In addition, the Mississippi State Department of Health response to those cases is presented and discussed.

The Evolving Medical and Veterinary Importance of the Gulf Coast tick (Acari: Ixodidae).

Amblyomma maculatum Koch (the Gulf Coast tick) is a three-host, ixodid tick that is distributed throughout much of the southeastern and south-central United States, as well as several countries throughout Central and South America. A considerable amount of scientific literature followed the original description of A. maculatum in 1844; nonetheless, the Gulf Coast tick was not recognized as a vector of any known pathogen of animals or humans for >150 years. It is now identified as the principal vector of Hepatozoon americanum, the agent responsible for American canine hepatozoonosis, and Rickettsia parkeri, the cause of an emerging, eschar-associated spotted fever group rickettsiosis identified throughout much of the Western Hemisphere. Coincident with these discoveries has been recognition that the geographical distribution of A. maculatum in the United States is far more extensive than described 70 yr ago, supporting the idea that range and abundance of certain tick species, particularly those with diverse host preferences, are not fixed in time or space, and may change over relatively short intervals. Renewed interest in the Gulf Coast tick reinforces the notion that the perceived importance of a particular tick species to human or animal health can be relatively fluid, and may shift dramatically with changes in the distribution and abundance of the arthropod, its vertebrate hosts, or the microbial agents that transit among these organisms.

Comparison of Tick Feeding Success and Vector Competence for Borrelia burgdorferi Among Immature Ixodes scapularis (Ixodida: Ixodidae) of Both Southern and Northern Clades.

Northern and southern Ixodes scapularis Say populations differ greatly in density, host utilization, and especially questing behavior of the immatures. Haplotypes of I. scapularis in North America can be divided into two major clades-the All American Clade (haplotypes A through J) and the Southern Clade (M through O). This genetic variation may affect feeding success and vector competence. This study compared feeding success of larval I. scapularis measured by time-to-drop-off and subsequent transmissibility success of Borrelia burgdorferi to mice using ticks from Mississippi, Connecticut (both F haplotype), and Louisiana (haplotype O). Northern ticks (CT) fed to repletion much faster than MS and LA ticks: overall, 73.6% of CT ticks had dropped off mice at Day 3 compared to only 1.7% and 6.6% of ticks dropped off for MS and LA ticks at that same time point. As for vector competence, 4 of the 4 mice in each case (MS or CT) that had been fed on by infected nymphs tested positive for B. burgdorferi. In a second experiment, 5 of the 6 mice tested positive for B. burgdorferi after exposure to infected LA ticks as compared with 3 of the 4 mice exposed to infected CT ticks. These data demonstrate that there is no difference in northern and southern populations of I. scapularis in their ability to transmit B. burgdorferi, but the ability of the northern populations to feed rapidly on rodents exceeds that of southern populations.

Investigations into Outbreaks of Black Fly Attacks and Subsequent Avian Haemosporidians in Backyard-Type Poultry and Other Exposed Avian Species.

In late spring of 2009 and 2010, there were reports of severe black fly (Simulium spp., shown in Fig. 1) outbreaks in various counties in Mississippi, especially those in and around the Mississippi River Delta. Complaints were of black flies attacking multiple species of backyard poultry and causing high morbidity and mortality in affected flocks. At several affected locations, black flies were readily observed swarming around and feeding on birds. A large number of these parasites were easily trapped on fly strips (Fig. 2). Multifocal to coalescing cutaneous hemorrhagic lesions, consistent with fly bites, were seen on the birds. Upon necropsy examination, a large number of black flies were also observed in the digestive tract (Fig. 3). Although black flies may cause disease directly, such as cardiopulmonary collapse and anaphylactoid reactions, detection of Leucocytozoon in blood smears (Fig. 4) of affected birds prompted further investigations of this protozoan as a cause of disease. Leucocytozoon spp. are known to be transmitted by black flies and may be associated with morbidity and mortality in birds such as poultry. From June 2009 through July 2012, the investigation included a total collection of 1068 individual blood samples, representing 371 individual premises in 89 counties/parishes across Mississippi (59), Alabama (10), Louisiana (4), and Tennessee (16). Of the 371 premises where blood samples were collected, 96 (26%) were either positive or highly suspected to be positive for Leucocytozoon spp. by blood smear analysis, and 5 (1.2%) were positive for Haemoproteus spp. by blood smear analysis. Attempts to diagnose Leucocytozoon spp. by PCR analysis and sequencing were complicated by coinfections with two closely related haemosporidians (Haemoproteus spp. and Plasmodium spp.). A novel technique involving flow cytometry was also explored. This study discusses the black fly field outbreak, the involvement of haemosporidians, molecular methods for detection of both the black flies and blood parasites, and initial attempts at flow cytometry.

Abundance and Diversity of Mosquito Species Collected From a Rural Area of Central Mississippi: Implications for West Nile Virus Transmission in Mississippi.

To determine abundance and seasonality of potential West Nile virus (WNV) mosquito vectors in a forested area of central Mississippi, mosquitoes were collected weekly from a wildlife management area located approximately 10 mi from a local urban area known to have numerous human WNV cases. We were particularly interested in the presence or absence of Culex quinquefasciatus, the primary vector of WNV in Mississippi, although other Culex species were assayed. Two Centers for Disease Control and Prevention light traps baited with CO2 were set once a week from 2005 through 2006 in the Pearl River Wildlife Management Area (PRWMA), which consists of 6,925 acres primarily composed of bottomland hardwood forest with wetland areas. Traps were placed midafternoon and picked up the following morning. A total of 199,222 mosquitoes were collected during the 2-year study. No Cx. quinquefasciatus were collected throughout the entire study, although other health department surveys have indicated they are abundant just a few miles away. As for other potential WNV vectors, 1,325 (0.6%) Cx. nigripalpus, 1,804 (0.9%) Cx. restuans, and 6,076 (3.1%) Cx. salinarius were collected in the PRWMA over the 2-year period. These data suggest that Cx. quinquefasciatus is not usually found in remote forested environments, but is more associated with human habitation.

Laboratory studies of movement and microenvironment choices of engorged adult female Amblyomma maculatum (Acari: Ixodidae).

Microenvironmental factors affect ovipositional choices and behavior in ticks. In this study, engorged female Amblyomma maculatum Koch were released in an observation arena covered with garden soil. The arena was evenly split into wet and dry sides, each containing 5 different types of structures (totaling 10). Upon release, observations at particular time points were made over 2 days as to speed, distance, actual paths traveled, and ultimate site selection, presumably for oviposition. In addition, time-lapse videography was utilized to record the track of each individual tick. This scenario was replicated 3 times with different cohorts of ticks (n = 5 per replicate). Of the total 15 ticks released in the arena, all attained a final site selection by 24 h. These final sites were 7/15 (47%) edge of arena; 2/15 (13%) under bark; 2/15 (13%) open soil; 2/15 (13%) at or near release point; 1/15 (7%) tunnel with animal fur; and 1/15 (7%) tunnel with chicken feathers. At final site selection, 11/15 (73%) subsequently burrowed into the soil, 3 of which were completely buried. Time-lapse videography revealed that most ticks (80%) explored the arena in a "looping back" fashion. Overall, engorged Gulf Coast ticks moved at an average speed of 0.09 mm/s, and the total distance traversed by the ticks averaged 2.37 m.

Bed bug saliva causes release of monocytic inflammatory mediators: plausible cause of cutaneous bite reactions.

Bed bugs may cause mild to severe cutaneous reactions. We studied the ability of bed bug salivary extract (SGE) to induce inflammatory reactions responsible for cutaneous manifestations and found that SGE stimulated the production of several potent chemokines and cytokines from macrophages. Chemokines induced by SGE included those known to recruit eosinophils (eotaxin), attract neutrophils [interleukin (IL)-8] or induce their survival and proliferation (granulocyte colony-stimulating factor). Cytokines involved in cell-mediated immunity, including IL-7, IL-10 and IL-12, were also induced. Saliva of bed bugs contains protein and non-protein molecules that have pleotropic effects on macrophages, orchestrating the immune response in the skin after bed bug bites.

Envenomation From Flood-Related Fire Ant Rafting: A Cautionary Note.

BACKGROUND: Imported fire ants aggressively sting humans, leading to a variety of local and systemic effects. Fire ants display a behavioral adaptation to survive flooding, called "rafting," bringing humans into even more contact with fire ants. METHODS: To assess frequency of encounters with fire ant rafts, duck hunters on the website "Duck Hunter's Forum" were asked about their experiences with the ants while wading or boating in flooded areas. All members of the group received a brief explanation and asked to respond directly if they had had such an encounter. They were then asked to fill out a short 6-question survey. RESULTS: There were 2021 views of the thread about fire ant raft encounters, with 35 (1.7%) responses. Twenty-four (68.6%) said they had experienced encounters with fire ant rafts. Six responders described purposely or inadvertently physically touching the rafts with their body parts, boat, or a paddle, causing the ants to enter their boat or to climb directly onto the hunter(s). Five respondents were stung (number of stings from "a few" to 50), describing reactions to stings as "small pustules," "whelps," and "pimple-like, puss-filled whelps." CONCLUSIONS: These results confirm that fire ant rafts formed during flooding present a potential hazard to persons walking, wading, or boating in those areas. We suggest that these rafts are an under-appreciated health hazard. It is reasonable to assume that people in flooded areas occasionally contact these floating rafts and are stung multiple times. We provide preliminary prevention/protection recommendations for health professionals, to be shared with their patients and the general public.

Density-dependent within-patch movement behavior of two competing species.

Movement behavior is central to understanding species distributions, population dynamics and coexistence with other species. Although the relationship between conspecific density and emigration has been well studied, little attention has been paid to how interspecific competitor density affects another species' movement behavior. We conducted releases of two species of competing Tribolium flour beetles at different densities, alone and together in homogeneous microcosms, and tested whether their recaptures-with-distance were well described by a random-diffusion model. We also determined whether mean displacement distances varied with the release density of conspecific and heterospecific beetles. A diffusion model provided a good fit to the redistribution of T. castaneum and T. confusum at all release densities, explaining an average of >60% of the variation in recaptures. For both species, mean displacement (directly proportional to the diffusion rate) exhibited a humped-shaped relationship with conspecific density. Finally, we found that both species of beetle impacted the within-patch movement rates of the other species, but the effect depended on density. For T. castaneum in the highest density treatment, the addition of equal numbers of T. castaneum or T. confusum had the same effect, with mean displacements reduced by approximately one half. The same result occurred for T. confusum released at an intermediate density. In both cases, it was total beetle abundance, not species identity that mattered to mean displacement. We suggest that displacement or diffusion rates that exhibit a nonlinear relationship with density or depend on the presence or abundance of interacting species should be considered when attempting to predict the spatial spread of populations or scaling up to heterogeneous landscapes.

Anomalous phenology of Ixodes scapularis (Ixodida: Ixodidae) in Mississippi.

The blacklegged tick, Ixodes scapularis Say, is one of the most important vectors of tick-borne disease agents in North America. Accordingly, it is crucial to know this species local composition, abundance, and seasonality (phenology) to help prevent tick-borne illnesses. The phenology of adult I. scapularis is reported in the scientific literature from October through May. Data from previous research in Mississippi have all supported this time frame for adult blacklegged tick activity. However, in this study, we report a collection of 13 I. scapularis from 9 widely separated areas in Mississippi during the summer and early fall of 2022 (June, July, and September). These findings are remarkable, even enigmatic, and should prompt further investigation.

Artificial infection of the bed bug with Rickettsia parkeri.

Although a variety of disease agents have been reported from bed bugs, the mechanical and biological disease transmission potential of bed bugs remains unelucidated. In this study we assayed survivability of the mildly pathogenic spotted fever group rickettsia, Rickettsia parkeri, in bed bugs after feeding on R. parkeri-infected chicken blood. Two groups of 15 adult bed bugs each were fed on infected or noninfected blood, and two groups of fourth-instar bed bugs also were fed on either infected or noninfected blood. One group of 15 adult bed bugs received no bloodmeal and was included as an additional control. Two weeks postfeeding, two pools of five live bed bugs from each group were surface sterilized, macerated, and placed in Vero cell cultures in an attempt to grow live organism. The remaining five individual bed bugs from each group were dissected, their salivary glands were removed for immunofluorescence assay (IFA) staining, and the remaining body parts were processed for polymerase chain reaction (PCR) analysis. Results indicated that no immature (now molted to fifth instar) bed bugs were positive for R. parkeri by IFA or PCR, indicating that organisms did not survive the molting process. After 4 wk of cell culture, no organisms were seen in cultures from any of the treatment or control groups, nor were any cultures PCR positive. However, two of the adult bed bugs were IFA positive for rickettsia-like organisms, and these two specimens were also PCR positive using R. parkeri-specific primers. These IFA and PCR results indicate that remnants of Rickettsia parkeri (possibly whole organisms) survived in the bugs for 2 wk, but the viability of the organisms in these two specimens could not be determined.