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Genetic characterization of Hawaiian isolates of Plasmodium relictum reveals mixed-genotype infections

Background: The relatively recent introduction of a highly efficient mosquito vector and an avian pathogen (Plasmodium relictum) to an isolated island ecosystem with nai??ve, highly susceptible avian hosts provides a unique opportunity to investigate evolution of virulence in a natural system. Mixed infections can significantly contribute to the uncertainty in host-pathogen dynamics with direct impacts on virulence. Toward further understanding of how host-parasite and parasite-parasite relationships may impact virulence, this study characterizes within-host diversity of malaria parasite populations based on genetic analysis of the trap (thrombospondin-related anonymous protein) gene in isolates originating from Hawaii, Maui and Kauai Islands. Methods: A total of 397 clones were produced by nested PCR amplification and cloning of a 1664 bp fragment of the trap gene from two malarial isolates, K1 (Kauai) and KV115 (Hawaii) that have been used for experimental studies, and from additional isolates from wild birds on Kauai, Maui and Hawaii Islands. Diversity of clones was evaluated initially by RFLP-based screening, followed by complete sequencing of 33 selected clones. Results: RFLP analysis of trap revealed a minimum of 28 distinct RFLP haplotypes among the 397 clones from 18 birds. Multiple trap haplotypes were detected in every bird evaluated, with an average of 5.9 haplotypes per bird. Overall diversity did not differ between the experimental isolates, however, a greater number of unique haplotypes were detected in K1 than in KV115. We detected high levels of clonal diversity with clear delineation between isolates K1 and KV115 in a haplotype network. The patterns of within-host haplotype clustering are consistent with the possibility of a clonal genetic structure and rapid within-host mutation after infection. Conclusion: Avian malaria (P. relictum) and Avipoxvirus are the significant infectious diseases currently affecting the native Hawaiian avifauna. This study shows that clonal diversity of Hawaiian isolates of P. relictum is much higher than previously recognized. Mixed infections can significantly contribute to the uncertainty in host-pathogen dynamics with direct implications for host demographics, disease management strategies, and evolution of virulence. The results of this study indicate a widespread presence of multiple-genotype malaria infections with high clonal diversity in native birds of Hawaii, which when coupled with concurrent infection with Avipoxvirus, may significantly influence evolution of virulence. ?? 2008 Jarvi et al; licensee BioMed Central Ltd.

Biology Direct

Tick control: Trapping, bio-control, host management and other alternative strategies

Biology of Ticks is the most comprehensive work on tick biology and tick-borne diseases. This second edition is a multi-authored work, featuring the research and analyses of renowned experts across the globe. Spanning two volumes, the book examines the systematics, biology, structure, ecological adaptations, evolution, genomics and the molecular processes that underpin the growth, development and survival of these important disease-transmitting parasites. Also discussed is the remarkable array of diseases transmitted (or caused) by ticks, as well as modern methods for their control. This book should serve as a modern reference for students, scientists, physicians, veterinarians and other specialists. Volume II includes chapters on the ecology of non-nidicolous and nidicolous ticks, genetics and genomics (including the genome of the Lyme disease vector Ixodes scapularis) and immunity, including host immune responses to tick feeding and tick-host interactions, as well as the tick's innate immune system that prevents and/or controls microbial infections. Six chapters cover in depth the many diseases caused by the major tick-borne pathogens, including tick-borne protozoa, viruses, rickettsiae of all types, other types of bacteria (e.g., the Lyme disease agent) and diseases related to tick paralytic agents and toxins. The remaining chapters are devoted to tick control using vaccines, acaricides, repellents, biocontrol, and, finally, techniques for breeding ticks in order to develop tick colonies for scientific study.

Book chapter

Pathogenicity of avian malaria in experimentally-infected Hawaii Amakihi

The introduction of avian malaria (Plasmodium relictum) and mosquitoes (Culex quinquefasciatus) to the Hawaiian Islands (USA) is believed to have played a major role in the decline and extinction of native Hawaiian honeycreepers (Drepanidinae). This introduced disease is thought to be one of the primary factors limiting recovery of honeycreepers at elevations below 1,200 m where native forest habitats are still relatively intact. One of the few remaining species of honeycreepers with a wide elevational distribution is the Hawaii Amakihi (Hernignathus virens). We measured morbidity and mortality in experimentally-infected Hawaii Amakihi that were captured in a high elevation, xeric habitat that is above the current range of the mosquito vector. Mortality among amakihi exposed to a single infective mosquito bite was 65% (13/20). All infected birds had significant declines in food consumption and a corresponding loss in body weight over the 60 day course of the experiment. Gross and microscopic lesions in birds that succumbed to malaria included enlargement and discoloration of the spleen and liver and parasitemias as high as 50% of circulating erythrocytes. Mortality in experimentally-infected amakihi was similar to that observed in Apapane (Himnatione sanguinea) and lower than that observed in Iiwi (Vestiaria coccinea) infected under similar conditions with the same parasite isolate. We conclude that the current elevational and geographic distribution of Hawaiian honeycreepers is determined by relative susceptibility to avian malaria.

Hawaii

Complex life-cycles in trophically transmitted helminths: Do the benefits of increased growth and transmission outweigh generalism and complexity costs?

Why do so many parasitic worms have complex life-cycles? A complex life-cycle has at least two hypothesized costs: (i) worms with longer life-cycles, i.e. more successive hosts, must be generalists at the species level, which might reduce lifetime survival or growth, and (ii) each required host transition adds to the risk that a worm will fail to complete its life-cycle. Comparing hundreds of trophically transmitted acanthocephalan , cestode , and nematode species with different life-cycles suggests these costs are weaker than expected. Helminths with longer cycles exhibit higher species-level generalism without impaired lifetime growth. Further, risk in complex life-cycles is mitigated by increasing establishment rates in each successive host. Two benefits of longer cycles are transmission and production. Longer cycles normally include smaller (and thus more abundant) first hosts that are likely to consume parasite propagules, as well as bigger (and longer-lived) definitive hosts, in which adult worms grow to larger and presumably more fecund reproductive sizes. Additional factors, like host immunity or dispersal, may also play a role, but are harder to address. Given the ubiquity of complex life-cycles, the benefits of incorporating or retaining hosts in a cycle must often exceed the costs.

Current Research in Parasitology & Vector-borne Di

Preliminary survey for entomopathogenic fungi associated with Ixodes scapularis (Acari: Ixodidae) in southern New York and New England, USA

Free-living larval, nymphal, and adult Ixodes scapularis Say were collected from scattered locales in southern New England and New York to determine infection rates with entomopathogenic fungi. Infection rates of larvae, nymphs, males, and females were 0% (571), 0% (272), 0% (57), and 4.3% (47), respectively. Two entomopathogenic fungi were isolated from field-collected I. scapularis females from Fire Island, NY. Isolates were identified as Verticillium lecanii (Zimmermann) Viegas and Verticillium sp. (a member of the Verticillium lecanii species complex). Ixodes scapularis Say is the principal vector of Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner ( Burgdorfer et al. 1982 , Johnson et al. 1984 ), the etiologic agent of Lyme disease in the northeastern and upper-midwestern United States. Control of I. scapularis is based on chemical treatment ( Mather et al. 1987b ; Schulze et al. 1987 , 1991 ), environmental management ( Wilson et al. 1988 , Schulze et al. 1995 ), and habitat modification ( Wilson 1986 ). These methods have shown variable success, and some potentially have negative environmental effects ( Wilson and Deblinger 1993 , Ginsberg 1994 ). Studies concerning natural predators, parasitoids, and pathogens of I. scapularis are rare. The use of ground-dwelling birds as tick predators has had only limited success ( Duffy et al. 1992 ). Nymphal I. scapularis are often infected with the parasitic wasp Ixodiphagus hookeri (Howard) ( Mather et al. 1987a , Hu et al. 1993 , Stafford et al. 1996 , Hu and Hyland 1997 ), but this wasp does not effectively control I. scapularis populations ( Stafford et al. 1996 ). The entomopathogenic nematodes Steinernema carpocapsae (Weiser) and S. glaseri (Steiner) are pathogenic only to engorged female I. scapularis, and thus have limited applicability ( Zhioua et al. 1995 ). In contrast, the entomogenous fungus Metarhizium anisopliae (Metschnikoff) Sorokin is highly pathogenic to all stages of I. scapularis, unfed as well as engorged, and thus has considerable potential as a microbial control agent ( Zhioua et al. 1997 ). European studies have suggested that entomopathogenic fungi might serve as natural controls of of Ixodes ricinus L. populations ( Samsinakova et al. 1974 , Eilenberg et al. 1991 , Kalsbeek et al. 1995 ). In the current study, we describe the isolation of entomopathogenic fungi from field-collected I. scapularis.

Journal of Medical Entomology

Detection of Splendidofilaria sp. (Onchocercidae:Splendidofilariinae) Microfilaria within Alaskan ground-dwelling birds in the grouse subfamily tetraoninae using taqman probe-based real-time PCR

Grouse and ptarmigan (Galliformes) harbor fairly diverse helminth faunas that can impact the host's health, including filarial nematodes in the genus Splendidofilaria . As host and parasite distributions are predicted to shift in response to recent climate change, novel parasites may be introduced into a region and impose additional stressors on bird populations. Limited information is available on the prevalence of filariasis in Alaska galliforms. To date, no molecular surveys have been completed. Past studies relied on examining blood smears or total body necropsies, which are time-consuming and may not detect filarial parasites with low prevalence in hosts. Therefore, we developed a TaqMan probe-based real-time PCR assay targeting the cytochrome c oxidase 1 gene ( COI ) of Splendidofilaria to decrease processing times and increase sensitivity as well as provide baseline data on the diversity of filariid infections in galliform species in Alaska. We screened a combined total of 708 galliform samples (678 unique individual birds) from different tissues (blood, muscle, and lung) for the presence of filarial DNA across the state of Alaska. Real-time PCR screening revealed an overall prevalence of filarial infection of 9.5% across species: Bonasa umbellus (0%, n = 23), Dendragapus fuliginosus (0%, n = 8), Falcipennis canadensis (26.8%, n = 198), Lagopus lagopus (2.6%, n = 274), Lagopus leucura (0%, n = 23), Lagopus muta (3%, n = 166), and Tympanuchus phasianellus (12.5%, n = 16). We observed microfilarial infections throughout most of Alaska except in Arctic regions and the Aleutian Islands where viable vectors may not be present.

Journal of Parasitology

Plague transforms positive effects of precipitation on prairie dogs to negative effects

Rodents characteristically benefit from increased precipitation, especially in typically dry habitats; “good years” of high precipitation improve their forage and water balance. However, Yersinia pestis (plague), a flea-borne pathogen of mammals that was introduced to western North America, has the greatest negative impact on at least some species of rodents during years of above-average precipitation. In the absence of plague mitigation, negative effects of plague in wet years might overwhelm the otherwise beneficial effects of increased moisture. In Montana and Utah, USA, where plague now occurs enzootically, we investigated the influence of precipitation on finite rates of annual population change (2000–2005) for 3 species of prairie dogs ( Cynomys spp.) in replicated plots treated with deltamethrin dust and in non-treated plots for paired comparisons. There was a significant interaction between precipitation and treatment. When we reduced plague vector fleas, prairie dog visual counts tended to increase with increasing precipitation. Simultaneously, there was a negative relationship between counts and precipitation on paired plots where plague was not managed, suggesting that plague transformed and reversed the otherwise beneficial effect of increased precipitation. Are the good years gone for prairie dogs? Even if the good years are not gone, they are perhaps relatively scarce compared to historic times prior to the invasion of plague. This scenario might apply to other ecosystems and may pose broad conservation challenges in western North America.

Montana, Utah

Understanding uncertainty in temperature effects on vector-borne disease: a Bayesian approach

Extrinsic environmental factors influence the distribution and population dynamics of many organisms, including insects that are of concern for human health and agriculture. This is particularly true for vector-borne infectious diseases like malaria, which is a major source of morbidity and mortality in humans. Understanding the mechanistic links between environment and population processes for these diseases is key to predicting the consequences of climate change on transmission and for developing effective interventions. An important measure of the intensity of disease transmission is the reproductive number R 0 . However, understanding the mechanisms linking R 0 and temperature, an environmental factor driving disease risk, can be challenging because the data available for parameterization are often poor. To address this, we show how a Bayesian approach can help identify critical uncertainties in components of R 0 and how this uncertainty is propagated into the estimate of R 0 . Most notably, we find that different parameters dominate the uncertainty at different temperature regimes: bite rate from 15°C to 25°C; fecundity across all temperatures, but especially ~25–32°C; mortality from 20°C to 30°C; parasite development rate at ~15–16°C and again at ~33–35°C. Focusing empirical studies on these parameters and corresponding temperature ranges would be the most efficient way to improve estimates of R 0 . While we focus on malaria, our methods apply to improving process-based models more generally, including epidemiological, physiological niche, and species distribution models.

Ecology

Ecology and management of plague in diverse communities of rodents and fleas

Plague originated in Asia as a flea-borne zoonosis of mammalian hosts. Today, the disease is distributed nearly worldwide. In western United States of America, plague is maintained, transmitted, and amplified in diverse communities of rodents and fleas. We examined flea diversity on three species of prairie dogs ( Cynomys spp., PDs) and six species of sympatric small rodents in Montana and Utah, United States of America. Among 2896 fleas, 19 species were identified; 13 were found on PDs and 9 were found on small rodents. In Montana, three flea species were found on PDs; the three species parasitize PDs and mice. In Utah, 12 flea species were found on PDs; the 12 species parasitize PDs, mice, voles, chipmunks, ground squirrels, rock squirrels, and marmots. Diverse flea communities and their willingness to parasitize many types of hosts, across multiple seasons and habitats, may favor plague maintenance and transmission. Flea parasitism on Peromyscus deer mice varied directly with elevation. Fleas are prone to desiccation, and might prosper at higher, mesic elevations; in addition, Peromyscus nest characteristics may vary with elevation. Effective management of plague is critical. Plague management is probably most effective when encompassing communities of rodents and fleas. Treatment of PD burrows with 0.05% deltamethrin dust, which suppressed fleas on PDs for >365 days, suppressed fleas on small rodents for at least 58 days. At one site, deltamethrin suppressed fleas on small rodents for at least 383 days. By simultaneously suppressing fleas on PDs and small rodents, deltamethrin should promote ecosystem resilience and One Health objectives.

Montana, Utah

Analysis of regional scale risk to whirling disease in populations of Colorado and Rio Grande cutthroat trout using Bayesian belief network model

Introduction and spread of the parasite Myxobolus cerebralis , the causative agent of whirling disease, has contributed to the collapse of wild trout populations throughout the intermountain west. Of concern is the risk the disease may have on conservation and recovery of native cutthroat trout. We employed a Bayesian belief network to assess probability of whirling disease in Colorado River and Rio Grande cutthroat trout ( Oncorhynchus clarkii pleuriticus and Oncorhynchus clarkii virginalis , respectively) within their current ranges in the southwest United States. Available habitat (as defined by gradient and elevation) for intermediate oligochaete worm host, Tubifex tubifex , exerted the greatest influence on the likelihood of infection, yet prevalence of stream barriers also affected the risk outcome. Management areas that had the highest likelihood of infected Colorado River cutthroat trout were in the eastern portion of their range, although the probability of infection was highest for populations in the southern, San Juan subbasin. Rio Grande cutthroat trout had a relatively low likelihood of infection, with populations in the southernmost Pecos management area predicted to be at greatest risk. The Bayesian risk assessment model predicted the likelihood of whirling disease infection from its principal transmission vector, fish movement, and suggested that barriers may be effective in reducing risk of exposure to native trout populations. Data gaps, especially with regard to location of spawning, highlighted the importance in developing monitoring plans that support future risk assessments and adaptive management for subspecies of cutthroat trout.

Arizona, Colorado, New Mexico, Utah, Wyoming

Borreliosis transmission from ticks to humans associated with desert tortoise burrows: Examples of tick-borne relapsing fever in the Mojave Desert

Ticks transmit pathogens and parasitize wildlife in turn causing zoonotic diseases in many ecosystems. Argasid ticks, such as Ornithodoros spp., harbor and transmit Borrelia spp., resulting in tick-borne relapsing fever (TBRF) in people. In the western United States, TBRF is typically associated with the bite of an infected Ornithodoros hermsi tick found in habitats at high elevations (>1500 ft). This report describes the first TBRF cases in people in the Mojave Desert (Clark County, NV). Individuals documented in these case studies were exposed to Ornithodoros ticks during excavation of soil burrows associated with Mojave Desert tortoises ( Gopherus agassizii ), with bacteria from one of the human case's blood sample genetically matching to Borrelia turicatae as determined by quantitative PCR and sequencing. Our findings should serve as a precaution to individuals working with tortoises or animal burrows, or those in contact with Ornithodoros ticks in this region.

California, Nevada

Using occupancy models to investigate the prevalence of ectoparasitic vectors on hosts: an example with fleas on prairie dogs

Ectoparasites are often difficult to detect in the field. We developed a method that can be used with occupancy models to estimate the prevalence of ectoparasites on hosts, and to investigate factors that influence rates of ectoparasite occupancy while accounting for imperfect detection. We describe the approach using a study of fleas (Siphonaptera) on black-tailed prairie dogs ( Cynomys ludovicianus ). During each primary occasion (monthly trapping events), we combed a prairie dog three consecutive times to detect fleas (15 s/combing). We used robust design occupancy modeling to evaluate hypotheses for factors that might correlate with the occurrence of fleas on prairie dogs, and factors that might influence the rate at which prairie dogs are colonized by fleas. Our combing method was highly effective; dislodged fleas fell into a tub of water and could not escape, and there was an estimated 99.3% probability of detecting a flea on an occupied host when using three combings. While overall detection was high, the probability of detection was always <1.00 during each primary combing occasion, highlighting the importance of considering imperfect detection. The combing method (removal of fleas) caused a decline in detection during primary occasions, and we accounted for that decline to avoid inflated estimates of occupancy. Regarding prairie dogs, flea occupancy was heightened in old/natural colonies of prairie dogs, and on hosts that were in poor condition. Occupancy was initially low in plots with high densities of prairie dogs, but, as the study progressed, the rate of flea colonization increased in plots with high densities of prairie dogs in particular. Our methodology can be used to improve studies of ectoparasites, especially when the probability of detection is low. Moreover, the method can be modified to investigate the co-occurrence of ectoparasite species, and community level factors such as species richness and interspecific interactions.

International Journal for Parasitology: Parasites

Comparison of flea sampling methods and Yersinia pestis detection on prairie dog colonies

Scientists collect fleas (Siphonaptera) to survey for Yersinia pestis , the bacterial agent of plague. When studying fleas parasitizing prairie dogs ( Cynomys spp.), two primary methods are used: (1) combing fleas from live-trapped prairie dogs and (2) swabbing fleas from burrows with cloth swabs attached to metal cables. Ideally, burrow swabbing, the cheaper and easier method, would explain flea burdens on prairie dogs and provide reliable information on plague prevalence. In a linear regression analysis of data from 1-month intervals (June–August 2010–2011) on 13 colonies of black-tailed prairie dogs ( Cynomys ludovicianus , BTPDs) in New Mexico, flea abundance on swabs explained 0–26% of variation in BTPD flea burdens. In an analysis of data (May–August 2016) from six colonies of BTPDs in Montana, flea abundance on swabs explained 2% of variation in BTPD flea burdens. In an analysis of data from a short-term interval (July 23–27, 2019) on four colonies of BTPDs in Montana, flea abundance on swabs explained 0.1% of variation in BTPD flea burdens. In an analysis of data from 1-week intervals (August–October 2000) on four colonies of white-tailed prairie dogs ( Cynomys leucurus , WTPD) in Utah, swabbing data explained 0.1% of variation in WTPD flea burdens. Pools of fleas from two WTPD colonies were tested for Y. pestis by mouse inoculation and isolation; 65% from WTPDs tested positive, whereas 4% from burrows tested positive. Data herein also show that results from burrow swabbing can misrepresent flea species composition and phenology on prairie dogs. Burrow swabbing is useful for some purposes, but limitations should be acknowledged, and accumulated data should be interpreted with caution.

Montana, New Mexico, Utah

Prairie dog responses to vector control and vaccination during an initial Yersinia pestis invasion

We evaluated the invasion of plague bacteria Yersinia pestis into a population of black-tailed prairie dogs (Cynomys ludovicianus; BTPDs) in South Dakota. We aimed to ascertain if Y. pestis invaded slowly or rapidly, and to determine if vector (flea) control or vaccination of BTPDs assisted in increasing survival rates. We sampled BTPDs in 2007 (before Y. pestis documentation), 2008 (year of confirmed invasion), and 2009 (after invasion). We estimated annual BTPD re-encounter rates on three 9-ha plots treated annually with deltamethrin dust for flea control and three 9-ha plots lacking dust. In 2007 and 2008, approximately half the adult BTPDs live-trapped were injected subcutaneously with either an experimental plague vaccine (F1–V fusion protein) or placebo formulation; the remaining individuals were not inoculated. From 2007 to 2009, we sampled 1559 BTPDs on 2542 occasions. During 2007–2008, the prevalence and intensity of fleas on BTPDs were 69–97% lower on the dusted vs. no dust plots. From 2007 to 2008, the annual re-encounter rate of non-inoculated BTPDs was 150% higher on the dusted vs. no dust plots. During the same interval on the dusted plots, the re-encounter rate was 55% higher for vaccinated adult female BTPDs vs. nonvaccinated adult females, but the annual re-encounter rate was 19% lower for vaccinated adult males. By late August 2008, BTPDs were nearly extirpated from the no dust plots. During 2007–2008 and 2008–2009 on the dusted plots, which persisted, the BTPD re-encounter rate was 41% higher for vaccinated vs. non-vaccinated adult females but 35% lower for vaccinated adult males. Yersinia pestis erupted with vigor as it invaded. Flea control enhanced BTPD survival but did not offer full protection. Flea control and F1–V vaccination seemed to have additive, positive effects on adult females. Annual re-encounter rates were reduced for vaccinated adult males; additional experimentation is needed to further evaluate this trend.

International Journal of Parasitology: Parasites a