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Goethert, H. K.

Publications and source records attributed to Goethert, H. K..

4 recordsLinked to original sources

Deer tick virus genotypes are perpetuated by different modes of transmission

Deer tick virus (DTV), or lineage II Powassan virus, is an emergent tick-borne encephalitis virus in North America. Survivors frequently sustain neurologic sequelae. Nationally reported cases have been increasing. DTV is thought to be maintained in nature by multiple modes including horizontal transmission (from viremic host to tick), cofeeding transmission (between ticks feeding nearby) and by transovarial transmission (female to progeny). Analysis of the relative importance of each mode has been hindered by low enzootic transmission. In 2021, Marthas Vineyard, Massachusetts experienced an epizootic that allowed us to probe the modes of transmission on the island. We detected virus in 7.8% of questing deer tick nymphs (161 of 2063) and in 0.3% of lone star nymphs (2 of 678). Infected ticks had a highly focal distribution; 56% of infected ticks derived from only 4 of 71 collection sites. Tick mitochondrial genome sequencing demonstrated that infected ticks were not more likely to be siblings than negative ticks and, therefore, were unlikely to have inherited the infection. Whole viral genome sequencing revealed the presence of 3 genotypes, 58% were type1, 0.6% type2, and 13.7% type3. Tick host bloodmeal identification analyses determined that nymphs infected with type1 were significantly associated with having fed on shrews (50 of 94 type1 ticks, odds ratio=2.3, p<0.001). This is consistent with shrews serving as a reservoir. Ticks infected with type3, however, had no host associations, consistent with infection acquired by cofeeding. It may be that local DTV genetic variation is shaped by transmission modes or host associations. ImportanceDeer tick virus (DTV; Powassan lineage II) is a tick-borne encephalitis virus that causes a rare zoonosis in North America. Cases have been increasingly reported within the last decade. Is the recent risk trend due to increased transmission? How this virus is perpetuated in nature is not well understood. We took advantage of a natural epizootic on Marthas Vineyard to probe how the ticks there had become infected. Using a combination of viral whole genome sequencing and bloodmeal remnant identification in ticks, we find that the mode of transmission varied by viral genotype. One genotype is associated with ticks that had fed on shrews, and another did not depend on a specific reservoir host. Host associations may drive genetic diversity of deer tick virus and thus local host population dynamics may influence zoonotic risk.

ecology↗

Focal persistence and phylodynamics of Heartland virus in Georgia

Heartland virus (HRTV) is an emerging tick-bone virus associated with severe illness in the U.S. There are large gaps in knowledge of HRTV diversity, evolution, and transmission due to a paucity of HRTV-positive samples and genome sequences. We identified a focal site of HRTV- positive Amblyomma americanum ticks in central Georgia and developed a novel multiplex- amplicon sequencing assay to generate full HRTV genome sequences. By screening over 21,000 field-collected ticks from 2021-2023, we identified six positive pools. Five were collected from the site in central Georgia where our group first detected HRTV-positive ticks in 2019, and one from a site in western Georgia approximately 175 km away. The HRTV genome sequences from Georgia were highly related, even across this distance and over five years. Reference HRTV genome sequences from across the U.S. were also geographically clustered. Time-scaled phylogenetic analysis suggested recent spread of HRTV in the U.S., with all available sequences sharing a common ancestor within the last 300 years, and sequences from Georgia sharing a common ancestor within the last 40 years. Our observed spatial clustering of HRTV and the high degree of genetic conservation in our persistent focus suggest the importance of small spatial dynamics in HRTV transmission ecology. Author SummaryHeartland virus (HRTV) was first discovered in humans in 2009 and has since caused over 60 cases of severe and fatal disease in the United States. HRTV is transmitted by the lone star tick, Amblyomma americanum, across the Southeast, East coast, and Midwest. Little information is known about how this virus circulates and changes across time and space due to a lack of genetic data. Here, we created a new procedure to generate more genetic sequence data for HRTV and collected over 21,000 ticks to screen for HRTV across three years in Georgia. We generated 6 new HRTV sequences and compared them to existing sequences from our group in Georgia, and across the country, finding evidence of regional clustering of HRTV and highly related HRTV across time in Georgia. Our analyses additionally found that this virus was likely introduced to the U.S. in the last 300 years. Our study provides new context and information in understanding the landscape and transmission of HRTV in the U.S.

genomics↗

Evidence for Powassan virus deletions and defective RNA in field collected ticks

Powassan virus (POWV) is a tick-borne flavivirus in the tick-borne encephalitis virus (TBEV) serogroup endemic to the United States, Canada, and parts of Russia. POWV remains an under-studied pathogen, despite the potential for serious and life-threatening neurologic complications following infection. While prior studies have characterized viral diversity due to single nucleotide polymorphisms, little is known about POWV recombination, defective RNAs (D-RNAs), and functional structural variants (SVs). Understanding POWV recombination in its natural vector can provide important insights into its replication and evolution. Thus, we analyzed POWV sequence data from 51 ticks collected from the Northeast United States to characterize deletion expression levels and patterns in naturally infected ticks, and we compared these results to single-passage isolates. We found that deletions were common in POWV RNA from ticks and that several areas of the genome were enriched for recombination junctions. Deletions were often associated with areas of microhomology. While most deletions were sample-specific, two major deletion archetypes were observed across multiple tick samples. The first consisted of small 19-50 base deletions in the methyltransferase domain of the ns5 RNA-dependent RNA-polymerase gene, resulting in a mixture of putative SVs and D-RNAs. The second consisted of approximately 1600 base deletions spanning the ns2a-ns3 genes, resulting in putative D-RNAs with abrogated viral protease function. Protease deletions were significantly enriched after one passage in baby hamster kidney cells despite a decrease in overall deletion expression. These results demonstrate the proclivity of POWV for recombination, with potential implications for immune evasion and persistence in ticks. IMPORTANCEPowassan virus is a tick-borne flavivirus that can cause serious, life-threatening neurological disease. Understanding how Powassan virus replicates and evolves within its tick vector may elucidate factors important in persistence, transmission, and human disease. Defective RNAs are replication-incompetent viral genomes generated through internal deletions, which have been associated with disease severity and persistent infection in other viruses but have not been described for Powassan virus. Here, we show that Powassan virus produces abundant defective RNAs in field-caught ticks, and that expression patterns of these defective RNAs changes after one passage in mammalian cells. Although the function of these defective RNAs remains unknown, this work establishes a critical framework for investigating the role of defective RNAs in Powassan virus replication and transmission.

genomics↗

Zoonotic Babesia microti in the northeastern U.S.: evidence for the expansion of a specific parasite lineage

The recent range expansion of human babesiosis in the northeastern United States, once found only in restricted coastal sites, is not well understood. This study sought to utilize a large number of samples to examine the population structure of the parasites on a fine scale to provide insights into the mode of emergence across the region. 228 B. microti samples collected in endemic northeastern U.S. sites were genotyped using published VNTR markers. The genetic diversity and population structure were analysed on a geographic scale using Phyloviz and TESS. Three distinct populations were detected in northeastern US, each dominated by a single ancestral type. In contrast to the limited range of the Nantucket and Cape Cod populations, the mainland population dominated from New Jersey eastward to Boston. Ancestral populations of B. microti were sufficiently isolated to differentiate into distinct populations. Despite this, a single population was detected across a large geographic area of the northeast that historically had at least 3 distinct foci of transmission, central New Jersey, Long Island and southeastern Connecticut. We conclude that a single B. microti genotype has expanded across the northeastern U.S. The biological attributes associated with this parasite genotype that have contributed to such a selective sweep remain to be identified.\n\nAuthor summaryBabesiosis is a disease caused by a protozoan parasite, Babesia microti, related to malaria. The disease is acquired by the bite of the deer tick, the same tick that transmits Lyme disease. Although Lyme disease rapidly emerged over a wide range within the last 40 years, babesiosis remained rare with an extremely focal distribution. Within the last decade, the number of reports of babesiosis cases has increased from an expanded area of risk, particularly across the mainland of southern New England. We determined whether the expanded risk may be due to local intensification of transmission as opposed to introduction of the parasite. Historical fragmentation of the landscape suggests that sites of B. microti transmission should have been isolated and thus evidence of multiple genetically distinct populations should be found. By a genetic fingerprinting method, we found that samples from the new mainland sites were all genetically similar. We conclude that one parasite genetic lineage has recently expanded its distribution and now dominates, suggesting that it has some phenotypic attribute that may confer a selective advantage over others.

epidemiology↗