Single Mowing Event Does Not Reduce Abundance of Ixodes scapularis (Acari: Ixodidae) and Dermacentor variabilis (Acari: Ixodidae) on Recreational Hiking Trails.

Nuisance and medically important ticks can be abundant in a variety of forested landscapes, including recreational parks. Strategies to reduce the abundance of host-seeking ticks in high-use areas within parks are limited. Mowing vegetation is a recommended method to control ticks, but few studies have evaluated the efficacy of this practice. The goal of this study was to determine if a single mowing event could reduce the abundance of host-seeking ticks on recreational trails. Ticks were collected by dragging trails at three recreational parks in northern Minnesota during June and July, 2021. A pre-intervention sample was taken followed by six consecutive weeks of tick sampling. We encountered a total of 3,456 ticks (2,459 Ixodes scapularis Say and 997 Dermacentor variabilis L.) during the 7-week study period. There were no significant differences in the abundance of I. scapularis (adults) or D. variabilis (adults only) between control and mown trail sections. Mowing was a significant predictor of nymphal I. scapularis abundance but trended towards more ticks in mown sections compared to controls. These results suggest that a single mowing intervention during early June is likely to be ineffective as a strategy to reduce the risk of human contacts with ticks on trails.

Field Evaluations of Three Botanically Inspired Repellents Against the Blacklegged Tick, Ixodes scapularis (Acari: Ixodidae).

Three compounds synthetically-derived from botanicals sources, ethyl perillyl carbonate, geranyl isovalerate, and citronellyl cyclobutane carboxylate, were tested for repellent activity against Ixodes scapularis Say in a field trial. Tick drags were treated with the compounds or with N, N-diethyl-m-toluamide (DEET) at high (0.25 mg/cm2) or low (0.16 mg/cm2) concentrations. Negative controls included untreated drags and drags treated with acetone, the carrier for all repellents. Freshly treated drags (within 20 min) were used to collect I. scapularis ticks at a county park in Wisconsin. To assess effectiveness, we measured tick encounter rates, detachment rate, and time to detachment. None of the repellent treatments, including DEET, resulted in significantly fewer encounters compared to both control treatments. However, the percentage of ticks that detached within 3 min was significantly higher on drags treated with repellents compared to controls. DEET was the most effective, repelling 69.7-87% of ticks by 3 min, but the effectiveness of the three test compounds was still high, with 42-87% of ticks detaching by 3 min. For time to detachment, there were no significant differences between DEET and the three test compounds. We conclude that these synthetically-derived repellents were effective against I. scapularis in a field trial and could be viable alternatives to DEET.

Inter-annual variation in prevalence of Borrelia burgdorferi sensu stricto and Anaplasma phagocytophilum in host-seeking Ixodes scapularis (Acari: Ixodidae) at long-term surveillance sites in the upper midwestern United States: Implications for public health practice.

The geographic range of the blacklegged tick, Ixodes scapularis, and its associated human pathogens have expanded substantially over the past 20 years putting an increasing number of persons at risk for tick-borne diseases, particularly in the upper midwestern and northeastern United States. Prevention and diagnosis of tick-borne diseases rely on an accurate understanding by the public and health care providers of when and where persons may be exposed to infected ticks. While tracking changes in the distribution of ticks and tick-borne pathogens provides fundamental information on risk for tick-borne diseases, metrics that incorporate prevalence of infection in ticks better characterize acarological risk. However, assessments of infection prevalence are more labor intensive and costly than simple measurements of tick or pathogen presence. Our objective was to examine whether data derived from repeated sampling at longitudinal sites substantially influences public health recommendations for Lyme disease and anaplasmosis prevention, or if more constrained sampling is sufficient. Here, we summarize inter-annual variability in prevalence of the agents of Lyme disease (Borrelia burgdorferi s.s.) and anaplasmosis (Anaplasma phagocytophilum) in host-seeking I. scapularis nymphs and adults at 28 longitudinal sampling sites in the Upper Midwestern US (Michigan, Minnesota, and Wisconsin). Infection prevalence was highly variable among sites and among years within sites. We conclude that monitoring infection prevalence in ticks aids in describing coarse acarological risk trends, but setting a fixed prevalence threshold for prevention or diagnostic decisions is not feasible given the observed variability and lack of temporal trends. Reducing repeated sampling of the same sites had minimal impact on regional (Upper Midwest) estimates of average infection prevalence; this information should be useful in allocating scarce public health resources for tick and tick-borne pathogen surveillance, prevention, and control activities.

Integrated Tick Management in South Central Wisconsin: Impact of Invasive Vegetation Removal and Host-Targeted Acaricides on the Density of Questing Ixodes scapularis (Acari: Ixodidae) Nymphs.

As tick-borne disease incidence increases and pathogens expand into new areas, the need for effective tick management strategies is paramount. In this 5-yr study (2014-2018) conducted in south central Wisconsin, we assessed whether an integrated tick management approach, deployed during peak tick activity (May-August), was more effective at reducing black-legged ticks (Ixodes scapularis Say (Ixodida: Ixodidae)), than individual interventions. Using a factorial design, invasive vegetation removal (Amur honeysuckle, Lonicera maackii Ruprecht (Dipsacales: Caprifoliaceae) and common buckthorn, Rhamnus cathartica Linnaeus (Rosales: Rhamnaceae)) was coupled with deployments of permethrin-treated cotton nesting materials (tick tubes) that target the white-footed mouse (Peromyscus leucopus Rafinesque (Rodentia: Cricetidae)). Results show that the probability of encountering a larval tick by drag sampling was unaffected by treatments at the cumulative 5-yr level. However, vegetation removal significantly reduced larval encounters in 2014, 2015, and 2018, by 33%, 57%, and 61% respectively, and reduced the density of questing nymphal (DON) ticks by 45% in 2015 compared to controls. Despite the limited effect on DON, vegetation removal significantly reduced the cumulative 5-yr density of Borrelia burgdorferi sensu stricto infected nymphs (DIN) (70%) compared to controls as a result of decreased nymphal infection prevalence. Sites treated with tick tubes had lower DIN (66%) and DON (54%) across the study and nymphs were reduced every year following the initial year of deployment compared to controls. Combining treatments did not further reduce DIN or DONs. We conclude that long-term integration of tick tubes with invasive vegetation removal does not provide additional benefit over individual treatments alone.

High Prevalence of Borrelia mayonii (Spirochaetales: Spirochaetaceae) in Field-Caught Tamias striatus (Rodentia: Sciuridae) From Northern Wisconsin.

Borrelia mayonii is a recently discovered bacterial spirochete that causes Lyme disease and is transmitted by the blacklegged tick, Ixodes scapularis Say (Acari: Ixodidae). To date, B. mayonii has been isolated from two vertebrate host species in Minnesota: field-caught white-footed mice (Peromyscus leucopus Rafinesque; Rodentia: Cricetidae) and American red squirrel (Tamiasciurus hudsonicus Erxleben). Here, we describe the first detection of B. mayonii in field-caught eastern chipmunks (Tamias striatus L. (Rodentia: Cricetidae)) from northern Wisconsin. During our study, we captured 530 unique small mammals and found an infection prevalence of 23.50% in field-caught eastern chipmunks (4/17) and 1.19% in Peromyscus spp. (5/420). Mean larval and nymphal burdens were determined for captured Blarina brevicauda (0, 0), Glaucomys volans (0.29, 0.14), Myodes gapperi (0.27, 0), Napaeozapus insignis (0, 0.25), Peromyscus spp. (1.88, 0.11), T. striatus (1.06, 0.65), and Sorex cinereus (0.09, 0). The high B. mayonii infection prevalence in eastern chipmunks suggests that the species may be an important reservoir for B. mayonii in the Upper Midwest.

Do-It-Yourself Tick Control: Granular Gamma-Cyhalothrin Reduces Ixodes scapularis (Acari: Ixodidae) Nymphs in Residential Backyards.

Lyme disease is the most common vector-borne disease in the United States with hotspots in the Northeast and Midwest. Integrated vector control for mosquito-borne disease prevention is often organized at the community level, but tick control is primarily coordinated at the household and individual level. Management of the blacklegged tick, Ixodes scapularis (Say), the vector of the causative agent of Lyme disease in the Midwest and eastern United States in peridomestic environments may be critical as many tick encounters are reported to occur in the yard. Therefore, we assessed the effectiveness of a widely available and low-cost pesticide that targets common lawn pests and is labeled for use against ticks. In June 2019, we evaluated a granular form of gamma-cyhalothrin in a placebo-controlled residential backyard study (n = 90) in two communities in Wisconsin. The product applied by the research team reduced nymphal blacklegged ticks in plots established in the lawn part of the ecotone by 97% one week after application at both communities and by 89-97% three to four weeks postapplication. The proportion of homes with at least one nymphal tick postapplication was significantly lower at acaricide-treated homes and ranged from 4.2 to 29.2% compared with placebo homes where at least one nymphal tick was found at 50-81.5% of homes. These results support the efficacy of a low-cost do-it-yourself strategy for homeowners seeking to reduce blacklegged ticks in the yard.

Immature Ixodes scapularis (Acari: Ixodidae) Collected From Peromyscus leucopus (Rodentia: Cricetidae) and Peromyscus maniculatus (Rodentia: Cricetidae) Nests in Northern Wisconsin.

The blacklegged tick, Ixodes scapularis Say, is the primary Lyme disease vector in the eastern United States. Both immature stages of I. scapularis take blood meals from mice belonging to the genus Peromyscus. Mice are active during the night and spend the majority of diel periods in nests. Thus, immature I. scapularis have a greater opportunity to drop from Peromyscus hosts while in nests compared with the forest floor. Here, we collected 11 Peromyscus nests during a 3-mo period during which the immature I. scapularis are known to be active. We then examined nesting materials for the presence of I. scapularis. Immature I. scapularis were detected in 64% of Peromyscus nests examined. Additionally, 55% of the nests contained at least one Dermacentor variabilis Say larva. Eighty-seven percent of all larval ticks found within nests were blood-fed. Because Peromyscus spp. are highly competent reservoirs of numerous tick-borne pathogens, the ticks that detach in their nests may be important for the maintenance of tick-borne diseases. However, further studies are needed to determine the fate of the I. scapularis that detach in Peromyscus nests.

Passive Animal Surveillance to Identify Ticks in Wisconsin, 2011-2017.

The introduction of new tick species poses a risk to human and animal health. Systematic active surveillance programs are expensive and uncommon. We evaluated a passive animal surveillance program as a monitoring tool to document the geographic distribution and host associations of ticks in Wisconsin. Passive surveillance partners included veterinary medical clinics, domestic animal shelters, and wildlife rehabilitation centers from 35 of the 72 Wisconsin counties. A total of 10,136 tick specimens were collected from 2325 animals from July 2011 to November 2017 and included Dermacentor variabilis Say (29.7% of all ticks), Ixodes texanus Banks (25.5%), Ixodes scapularis Say (19.5%), Haemaphysalis leporispalustris Packard (13.8%), Ixodes cookei Packard (4.4%), and Dermacentor albipictus Packard (1.7%). Less common species (<1% of collection) included Ixodes dentatus Marx, Ixodes sculptus Neumann, Ixodes marxi Banks, Amblyomma americanum Linnaeus, and Rhipicephalus sanguineus Latreille. Of the 2325 animals that were examined, most were domestic dogs (53%), eastern cottontail rabbits (16%), domestic cats (15%), and North American raccoons (11%). An additional 21 mammal and 11 bird species were examined at least once during the six years of the study. New county records are summarized for each species. Public health, academic, and veterinary and animal care partners formed a community of practice enabling effective statewide tick surveillance.

Ixodes scapularis does not harbor a stable midgut microbiome.

Hard ticks of the order Ixodidae serve as vectors for numerous human pathogens, including the causative agent of Lyme Disease Borrelia burgdorferi. Tick-associated microbes can influence pathogen colonization, offering the potential to inhibit disease transmission through engineering of the tick microbiota. Here, we investigate whether B. burgdorferi encounters abundant bacteria within the midgut of wild adult Ixodes scapularis, its primary vector. Through the use of controlled sequencing methods and confocal microscopy, we find that the majority of field-collected adult I. scapularis harbor limited internal microbial communities that are dominated by endosymbionts. A minority of I. scapularis ticks harbor abundant midgut bacteria and lack B. burgdorferi. We find that the lack of a stable resident midgut microbiota is not restricted to I. scapularis since extension of our studies to I. pacificus, Amblyomma maculatum, and Dermacentor spp showed similar patterns. Finally, bioinformatic examination of the B. burgdorferi genome revealed the absence of genes encoding known interbacterial interaction pathways, a feature unique to the Borrelia genus within the phylum Spirochaetes. Our results suggest that reduced selective pressure from limited microbial populations within ticks may have facilitated the evolutionary loss of genes encoding interbacterial competition pathways from Borrelia.

Prevalence of Tick-Borne Pathogens in Two Species of Peromyscus Mice Common in Northern Wisconsin.

Two species of mice, the white-footed mouse, Peromyscus leucopus (Rafinesque; Rodentia: Cricetidae) and the woodland deer mouse, Peromyscus maniculatus (Wagner; Rodentia: Cricetidae), serve as reservoirs of tick-borne pathogens in many parts of North America. However, the role P. maniculatus plays in the amplification and maintenance of Anaplasma phagocytophilum (Rickettsiales: Ehrlichiaceae) and Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) is not well understood. In northern Wisconsin, from 2012 to 2014, 560 unique mice were captured at 83 sites distributed across five forests. P. leucopus was more likely infested with immature Ixodes scapularis compared to P. maniculatus (60.1 vs. 28.3%). Abundance of immature I. scapularis on P. leucopus (M = 2.69; SD = 3.53) was surprisingly low and even lower for P. maniculatus (M = 0.717; SD = 1.44). Both P. leucopus and P. maniculatus were infected with B. burgdorferi, 24.0 and 16.8%, respectively. The prevalence of A. phagocytophilum infection in P. leucopus (1.69%) was similar to that observed in P. maniculatus (4.73%). Nine of 10 mice co-infected with both pathogens were P. maniculatus, and there were more co-infections in this species than expected by chance (3.07 vs. 0.82%). Differences in the behavior and biology between these two mice species may contribute to the variation observed in the abundance of host-attached ticks and pathogen prevalence. These differences highlight a potential hazard of the failure to differentiate between these visually similar mice, but there is evidence that these two mice species can each serve as reservoirs for tick-borne pathogens that cause human disease in Wisconsin.

Prevalence and Distribution of Human and Tick Infections with the Ehrlichia muris-Like Agent and Anaplasma phagocytophilum in Wisconsin, 2009-2015.

Ehrlichiosis and anaplasmosis are important emerging tickborne zoonoses that affect both humans and animals. Knowledge of the geographic distribution and prevalence of Ehrlichia spp. and Anaplasma phagocytophilum in Wisconsin is important information as a baseline for future comparisons. Reported human cases between 2009 and 2015 were identified using the Wisconsin Electronic Disease Surveillance System (WEDSS) and mapped by county of residence. Vector surveillance was established using ticks collected from animals by partners, including veterinary medical clinics, domestic animal shelters, and wildlife rehabilitation centers from 40 Wisconsin counties. A total of 1835 Ixodes scapularis tick specimens (larvae, nymphs, and adults) were collected from 18 different domestic and wildlife species from July 2011 to November 2015. An additional 1136 nymphs were collected by drag sampling at 23 locations in 19 counties in 2015. A real-time PCR assay that detects and distinguishes several Ehrlichia species, including a pathogenic Ehrlichia muris-like agent (EMLA), and A. phagocytophilum was performed on adult and nymphal ticks. A total of 757 I. scapularis ticks (predominately adults) were tested from animal collections, with 67 (8.9%) individuals positive for A. phagocytophilum and 22 (2.9%) positive for EMLA DNA. Of the 1150 questing nymphs, 62 (5.4%) were positive for A. phagocytophilum and 10 (0.9%) were positive for EMLA DNA. Specimens of I. scapularis that were positive for A. phagocytophilum were found in 27 of the 33 counties surveyed. Specimens that were positive for EMLA were less common and were found in nine counties. This study provides the first statewide survey of I. scapularis ticks for these pathogens and indicates that the risk of human exposure is widely distributed.

Occurrence of Amblyomma americanum (Acari: Ixodidae) and Human Infection With Ehrlichia chaffeensis in Wisconsin, 2008-2015.

Because of the increasing incidence of human ehrlichiosis in Wisconsin, we assessed reports of human infections by Ehrlichia chaffeensis and the distribution of its vector, the lone star tick (Amblyomma americanum (L.)). From 2008 through 2015, 158 probable and confirmed human cases of E. chaffeensis infections were reported to the Wisconsin Department of Health Services. Five cases without travel history outside of Wisconsin were confirmed as E. chaffeensis by polymerase chain reaction. Surveillance for the vector occurred from 2008 through 2015 and was based on active and passive methods, including examination of white-tailed deer, collections from live-trapped small mammals, submissions of ticks removed from wild and domestic animals through the Wisconsin Surveillance of Animals for Ticks (SWAT) program, digital or physical submissions by the public to the University of Wisconsin Insect Diagnostic or Medical Entomology laboratories, and active tick dragging. More than 50 lone star ticks (46 adults, 6 nymphs, and 1 larva) were identified. Lone star ticks were more commonly found in south central Wisconsin, particularly in Dane County, where discovery of more than one life stage in a single year indicates possible establishment.

Borrelia mayonii sp. nov., a member of the Borrelia burgdorferi sensu lato complex, detected in patients and ticks in the upper midwestern United States.

Lyme borreliosis (LB) is a multisystem disease caused by spirochetes in the Borrelia burgdorferisensu lato (Bbsl) genospecies complex. We previously described a novel Bbsl genospecies (type strain MN14-1420T) that causes LB among patients with exposures to ticks in the upper midwestern USA. Patients infected with the novel Bbsl genospecies demonstrated higher levels of spirochetemia and somewhat differing clinical symptoms as compared with those infected with other Bbsl genospecies. The organism was detected from human specimens using PCR, microscopy, serology and culture. The taxonomic status was determined using an eight-housekeeping-gene (uvrA, rplB, recG, pyrG, pepX, clpX, clpA and nifS) multi-locus sequence analysis (MLSA) and comparison of 16S rRNA gene, flaB, rrf-rrl, ospC and oppA2 nucleotide sequences. Using a system threshold of 98.3 % similarity for delineation of Bbsl genospecies by MLSA, we demonstrated that the novel species is a member of the Bbsl genospecies complex, most closely related to B. burgdorferisensu stricto (94.7-94.9 % similarity). This same species was identified in Ixodes scapularis ticks collected in Minnesota and Wisconsin. This novel species, Borrelia mayonii sp. nov, is formally described here. The type strain, MN14-1420, is available through the Deutsche Sammlung von Mikroorganismen und Zelkulturen GmbH (DSM 102811) and the American Type Culture Collection (ATCC BAA-2743).

Identification of a novel pathogenic Borrelia species causing Lyme borreliosis with unusually high spirochaetaemia: a descriptive study.

BACKGROUND: Lyme borreliosis is the most common tick-borne disease in the northern hemisphere. It is a multisystem disease caused by Borrelia burgdorferi sensu lato genospecies and characterised by tissue localisation and low spirochaetaemia. In this study we aimed to describe a novel Borrelia species causing Lyme borreliosis in the USA. METHODS: At the Mayo clinic, from 2003 to 2014, we tested routine clinical diagnostic specimens from patients in the USA with PCR targeting the oppA1 gene of B burgdorferi sensu lato. We identified positive specimens with an atypical PCR result (melting temperature outside of the expected range) by sequencing, microscopy, or culture. We collected Ixodes scapularis ticks from regions of suspected patient tick exposure and tested them by oppA1 PCR. FINDINGS: 100 545 specimens were submitted by physicians for routine PCR from Jan 1, 2003 to Sept 30, 2014. From these samples, six clinical specimens (five blood, one synovial fluid) yielded an atypical oppA1 PCR product, but no atypical results were detected before 2012. Five of the six patients with atypical PCR results had presented with fever, four had diffuse or focal rash, three had symptoms suggestive of neurological inclusion, and two were admitted to hospital. The sixth patient presented with knee pain and swelling. Motile spirochaetes were seen in blood samples from one patient and cultured from blood samples from two patients. Among the five blood specimens, the median oppA1 copy number was 180 times higher than that in 13 specimens that tested positive for B burgdorferi sensu stricto during the same time period. Multigene sequencing identified the spirochaete as a novel B burgdorferi sensu lato genospecies. This same genospecies was detected in ticks collected at a probable patient exposure site. INTERPRETATION: We describe a new pathogenic Borrelia burgdorferi sensu lato genospecies (candidatus Borrelia mayonii) in the upper midwestern USA, which causes Lyme borreliosis with unusually high spirochaetaemia. Clinicians should be aware of this new B burgdorferi sensu lato genospecies, its distinct clinical features, and the usefulness of oppA1 PCR for diagnosis. FUNDING: US Centers for Disease Control and Prevention Epidemiology and Laboratory Capacity for Infectious Diseases (ELC) Cooperative Agreement and Mayo Clinic Small Grant programme.

Detection of human pathogenic Ehrlichia muris-like agent in Peromyscus leucopus.

An Ehrlichia muris-like (EML) bacterium was recently detected in humans and Ixodes scapularis ticks in Minnesota and Wisconsin. The reservoir for this agent is unknown. To investigate the occurrence of the EML agent, groEL PCR testing and sequencing was performed on blood from small mammals and white-tailed deer that were collected in areas where human and tick infections were previously demonstrated. DNA of the EML agent was detected in two Peromyscus leucopus of 146 small mammals (1.4%); while 181 O. virginianus tested negative. This report provides the first evidence that DNA from the EML agent is found in P. leucopus, the same animal that is a reservoir for Anaplasma phagocytophilum in this region. The role of white-tailed deer remains inconclusive. Further sampling is warranted to understand the spatial and temporal distribution, transmission and maintenance of this pathogen.

Prevalence of Borrelia burgdorferi and Anaplasma phagocytophilum in Ixodes scapularis (Acari: Ixodidae) nymphs collected in managed red pine forests in Wisconsin.

Changes in the structure of managed red pine forests in Wisconsin caused by interacting root- and stem-colonizing insects are associated with increased abundance of the blacklegged tick, Ixodes scapularis Say, in comparison with nonimpacted stands. However, the frequency and variability of the occurrence of tick-borne pathogens in this coniferous forest type across Wisconsin is unknown. Red pine forests were surveyed from 2009 to 2013 to determine the prevalence of Borrelia burgdorferi and Anaplasma phagocytophilum in questing I. scapularis nymphs. Polymerase chain reaction analysis revealed geographical differences in the nymphal infection prevalence (NIP) of these pathogens in red pine forests. In the Kettle Moraine State Forest (KMSF) in southeastern Wisconsin, NIP of B. burgdorferi across all years was 35% (range of 14.5-53.0%). At the Black River State Forest (BRSF) in western Wisconsin, NIP of B. burgdorferi across all years was 26% (range of 10.9-35.5%). Differences in NIP of B. burgdorferi between KMSF and BRSF were statistically significant for 2010 and 2011 and for all years combined (P < 0.05). NIP ofA. phagocytophilum (human agent) averaged 9% (range of 4.6-15.8%) at KMSF and 3% (range of 0-6.4%) at BRSF, and was significantly different between the sites for all years combined (P < 0.05). Differences in coinfection of B. burgdorferi and A. phagocytophilum were not statistically significant between KMSF and BRSF, with an average of 3.4% (range of 1.7-10.5%) and 2.5% (range of 0-5.5%), respectively. In 2013, the density of infected nymphs in KMSF and BRSF was 14 and 30 per 1000m2, respectively, among the highest ever recorded for the state. Differences in the density of nymphs and NIP among sites were neither correlated with environmental factors nor time since tick colonization. These results document significant unexplained variation in tick-borne pathogens between coniferous forests in Wisconsin that warrants further study.

Hunter-killed deer surveillance to assess changes in the prevalence and distribution of Ixodes scapularis (Acari: Ixodidae) in Wisconsin.

As a result of the increasing incidence of Lyme disease and other tick-borne pathogens in Wisconsin, we assessed the distribution of adult blacklegged ticks through collections from hunter-killed deer in 2008 and 2009 and compared results with prior surveys beginning in 1981. Volunteers staffed 21 Wisconsin Department of Natural Resources registration stations in 21 counties in the eastern half of Wisconsin in 2008 and 10 stations in seven counties in northwestern Wisconsin in 2009. In total, 786 and 300 white-tailed deer (Odocoileus virginianus) were examined in 2008 and 2009, respectively. All but three stations in 2008 were positive for ticks and all stations in 2009 were positive for ticks. The three sites negative for ticks occurred within the eastern half of Wisconsin. The results indicate that range expansion of Ixodes scapularis (Say) is continuing and the risk of tick exposure is increasing, especially in the eastern one-third of the state.