Complete Mitochondrial Genome Sequence of the Gulf Coast Tick (Amblyomma maculatum).
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ABSTRACT:
SUBMITTER: Brenner AE
PROVIDER: S-EPMC8459658 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:BackgroundSaliva of blood sucking arthropods contains compounds that antagonize their hosts' hemostasis, which include platelet aggregation, vasoconstriction and blood clotting; saliva of these organisms also has anti-inflammatory and immunomodullatory properties. Perhaps because hosts mount an active immune response against these compounds, the diversity of these compounds is large even among related blood sucking species. Because of these properties, saliva helps blood feeding as well as help the establishment of pathogens that can be transmitted during blood feeding.Methodology/principal findingsWe have obtained 1,626,969 reads by pyrosequencing a salivary gland cDNA library from adult females Amblyomma maculatum ticks at different times of feeding. Assembly of this data produced 72,441 sequences larger than 149 nucleotides from which 15,914 coding sequences were extracted. Of these, 5,353 had >75% coverage to their best match in the non-redundant database from the National Center for Biotechnology information, allowing for the deposition of 4,850 sequences to GenBank. The annotated data sets are available as hyperlinked spreadsheets. Putative secreted proteins were classified in 133 families, most of which have no known function.Conclusions/significanceThis data set of proteins constitutes a mining platform for novel pharmacologically active proteins and for uncovering vaccine targets against A. maculatum and the diseases they carry.
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Project description:Geographic distribution of Rickettsia parkeri in its US tick vector, Amblyomma maculatum, was evaluated by PCR. R. parkeri was detected in ticks from Florida, Georgia, Kentucky, Mississippi, Oklahoma, and South Carolina, which suggests that A. maculatum may be responsible for additional cases of R. parkeri rickettsiosis throughout much of its US range.
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Project description:Borrelia spp. are agents of Lyme disease and relapsing fever, diseases which use Ixodes hard ticks and Ornithodoros soft ticks, respectively, as primary vectors. Some relapsing fever spirochetes, such as B. miyamotoi, are also found in hard ticks. To date, no Borrelia sp. is known to use the hard tick, Amblyomma maculatum, as a vector. However, both B. burgdorferi and B. lonestari were recently detected in A. maculatum removed from hosts. In our study, DNA extracts from 306 questing adult A. maculatum collected in Mississippi in 2009 and 2010 were tested for Borrelia spp. DNA by PCR amplification of flaB and 16S rRNA gene targets. An additional 97 A. maculatum collected in 2013 were tested by amplification of 16S rRNA gene target. Two ticks, one collected in 2009 and the other in 2010, were positive by PCR of the flaB and 16S rRNA gene targets; both were collected from the same location in central Mississippi. Interestingly, 16S rRNA gene amplicons from these two tick extracts were 98% identical to twelve Borrelia spp. including the reptile-associated spirochete B. turcica and Borrelia sp. "tAG158M"; flaB amplicons from these two ticks shared closest identity (89%) to the reptile-associated spirochete, B. turcica. These results demonstrate a Borrelia sp. in unfed A. maculatum ticks that is unique from other species in the NCBI database and in a clade with reptile-associated Borrelia species. Detection of a previously unrecognized Borrelia in a hard tick species generates additional questions regarding the bacterial fauna in these arthropods and warrants further studies to better understand this fauna.
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Project description:The Gulf Coast tick (Amblyomma maculatum) has evolved as a competent vector of the spotted-fever group rickettsia, Rickettsia parkeri. In this study, the functional role of catalase, an enzyme responsible for the degradation of toxic hydrogen peroxide, in the colonization of the tick vector by R. parkeri and transovarial transmission of this pathogen to the next tick generation, was investigated. Catalase gene (CAT) expression in midgut, salivary glands and ovarian tissues exhibited a 2-11-fold increase in transcription level upon R. parkeri infection. Depletion of CAT transcripts using an RNA-interference approach significantly reduced R. parkeri infection levels in midgut and salivary gland tissues by 53-63%. The role of CAT in transovarial transmission of R. parkeri was confirmed by simultaneously blocking the transcript and the enzyme by injecting double-stranded RNA for CAT and a catalase inhibitor (3-amino-1,2,4-triazole) into gravid females. Simultaneous inhibition of the CAT transcript and the enzyme significantly reduced the egg conversion ratio with a 44% reduction of R. parkeri transovarial transmission. These data suggest that catalase is required for rickettsial colonization of the tick vector and transovarial transmission to the next generation.
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Project description:Tick selenoproteins have been associated with antioxidant activity in ticks. Thioredoxin reductase (TrxR), also a selenoprotein, belongs to the pyridine nucleotide-disulphide oxidoreductase family of proteins and is an important antioxidant. Molecular interactions between native microbiota and tick hosts have barely been investigated to date. In this study, we determined the functional role of TrxR in tick feeding and in maintenance of the native microbial community. TrxR transcript levels remained high and microbial load was reduced throughout tick attachment to the vertebrate host. RNA interference (RNAi) showed that depletion of TrxR activity did not interfere with tick haematophagy or phenotype but did reduce the viability of the microbiome within the tick tissues, presumably by perturbing redox homeostasis. The transcriptional activity of various antioxidant genes remained unaffected whereas the antioxidant genes Manganese superoxide dismutase (MnSOD), copper/zinc superoxide dismutase (Cu/Zn SOD) and selenoprotein M (SelM) were significantly down-regulated in salivary glands of the ticks subjected to RNAi. The perturbed TrxR enzymatic activity in the knocked-down tick tissues negatively affected the bacterial load as well. Furthermore, we observed the altered bacterial profiles in TrxR-silenced tick tissues. Taken together, these results indicate an essential functional role for TrxR in maintaining the bacterial community associated with ticks.
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Project description:As obligate blood-sucking ectoparasites, to avoid tissue damage, ticks must neutralize the reactive oxygen species (ROS) generated from uptake and digestion of a bloodmeal. Consequently, ticks utilize a battery of antioxidant molecules, including catalase (CAT), an enzyme that converts hydrogen peroxide (H2 O2 ) into water and oxygen. Here, we investigated the tick antioxidant machinery by exogenous injection of sublethal doses of H2 O2 or paraquat. The relative transcript levels of selected Amblyomma maculatum antioxidant targets in tissues were determined by quantitative reverse transcriptase PCR following treatment. The results showed 2-16-fold increases in target antioxidant gene transcripts, signifying the ability of Am. maculatum to regulate its antioxidant machinery when exposed to increased ROS levels. Next, RNA interference was used to determine the functional role of CAT in haematophagy, redox homeostasis and reproductive fitness. CAT gene silencing was confirmed by transcript depletion within tick tissues; however, CAT knockdown alone did not interfere with tick haematophagy or phenotype, as confirmed by the resulting differential expression of antioxidant genes, thereby indicating an alternative mechanism for ROS control. Interestingly, double stranded RNA of CAT gene (dsCAT) and the CAT inhibitor, 3-aminotriazole, together reduced tick reproductive fitness via a marked reduction in egg mass and larval eclosion rates, highlighting a role for CAT in tick redox-homeostasis, making it a potential target for tick control.
| S-EPMC4860135 | biostudies-literature
Project description:Our objective of this study was to explore the bacterial microbiome in fresh or fresh-frozen adult Amblyomma maculatum (Gulf Coast ticks) using extracts enriched for microbial DNA. We collected 100 questing adult A. maculatum, surface disinfected them, and extracted DNA from individual ticks collected the same day or after storage at -80?°C. Because only extracts with microbial DNA concentrations above 2?ng/?L were considered suitable for individual analysis, we expected fewer samples to meet these requirements. Of individual ticks extracted, 48 extracts met this minimum concentration. We pooled 20 additional extracts that had lower concentrations to obtain seven additional pools that met the minimum DNA concentration. Libraries created from these 55 samples were sequenced using an Illumina MiSeq platform, and data sets were analyzed using QIIME to identify relative abundance of microorganisms by phylum down to genus levels. Proteobacteria were in greatest abundance, followed by Actinobacteria, Firmicutes, and Bacteroidetes, at levels between 1.9% and 6.4% average relative abundance. Consistent with the Francisella-like endosymbiont known to be present in A. maculatum, the genus Francisella was detected at highest relative abundance (72.9%; SE 0.02%) for all samples. Among the top ten genera identified (relative abundance???0.5%) were potential extraction kit contaminants, Sphingomonas and Methylobacterium, the soil bacterium Actinomycetospora, and the known A. maculatum-associated genus, Rickettsia. Four samples had Rickettsia at greater than 1% relative abundance, while nine additional samples had Rickettsia at low (0.01-0.04%) relative abundance. In this study, we used the entire microbe-enriched DNA extract for whole ticks for microbiome analysis. A direct comparison of the microbiome in microbe-enriched DNA and total genomic DNA extracts from halves of the same tick would be useful to determine the utility of this extraction method in this system. We anticipate that future tick microbiome studies will be valuable to explore the influence of microbial diversity on pathogen maintenance and transmission, and to evaluate niche-specific microbiomes within individual tick tissues.
| S-EPMC5962414 | biostudies-literature
Project description:The Gulf Coast tick, Amblyomma maculatum, is a vector of Rickettsia parkeri, a recently identified human pathogen that causes a disease with clinical symptoms that resemble a mild form of Rocky Mountain spotted fever. Because the prevalence of R. parkeri infection in geographically distinct populations of A. maculatum is not fully understood, A. maculatum specimens collected as part of a tick and pathogen surveillance system in Fairfax County, Virginia, were screened to determine pathogen infection rates. Overall, R. parkeri was found in 41.4% of the A. maculatum that were screened. Additionally, the novel spotted fever group Rickettsia sp., tentatively named "Candidatus Rickettsia andeanae," was observed for the first time in Virginia.
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Project description:Until recently, Amblyomma maculatum (the Gulf Coast tick) had garnered little attention compared to other species of human-biting ticks in the United States. A. maculatum is now recognized as the principal vector of Rickettsia parkeri, a pathogenic spotted fever group rickettsia (SFGR) that causes an eschar-associated illness in humans that resembles Rocky Mountain spotted fever. A novel SFGR, distinct from other recognized Rickettsia spp., has also been detected recently in A. maculatum specimens collected in several regions of the southeastern United States. In this study, 198 questing adult Gulf Coast ticks were collected at 4 locations in Florida and Mississippi; 28% of these ticks were infected with R. parkeri, and 2% of these were infected with a novel SFGR. Seventeen isolates of R. parkeri from individual specimens of A. maculatum were cultivated in Vero E6 cells; however, all attempts to isolate the novel SFGR were unsuccessful. Partial genetic characterization of the novel SFGR revealed identity with several recently described, incompletely characterized, and noncultivated SFGR, including "Candidatus Rickettsia andeanae" and Rickettsia sp. Argentina detected in several species of Neotropical ticks from Argentina and Peru. These findings suggest that each of these "novel" rickettsiae represent the same species. This study considerably expanded the number of low-passage, A. maculatum-derived isolates of R. parkeri and characterized a second, sympatric Rickettsia sp. found in Gulf Coast ticks.
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Project description:We report Rickettsia parkeri and Candidatus Rickettsia andeanae in ticks of the Amblyomma maculatum group collected from dogs in Sonora, Mexico. Molecular characterization of these bacteria was accomplished by DNA amplification and sequence analysis of portions of the rickettsial genes gltA, htrA, ompA, and ompB.
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