Phylogenetic diversity of two common Trypanosoma cruzi lineages in the Southwestern United States.

Trypanosoma cruzi is the causative agent of Chagas disease, a devastating parasitic disease endemic to Central and South America, Mexico, and the USA. We characterized the genetic diversity of Trypanosoma cruzi circulating in five triatomine species (Triatoma gerstaeckeri, T. lecticularia, T.indictiva, T. sanguisuga and T. recurva) collected in Texas and Southern Arizona using multilocus sequence typing (MLST) with four single-copy loci (cytochrome oxidase subunit II- NADH dehydrogensase subunit 1 region (COII-ND1), mismatch-repair class 2 (MSH2), dihydrofolate reductase-thymidylate synthase (DHFR-TS) and a nuclear gene with ID TcCLB.506529.310). All T. cruzi variants fall in two main genetic lineages: 75% of the samples corresponded to T. cruzi Discrete Typing Unit (DTU) I (TcI), and 25% to a North American specific lineage previously labelled TcIV-USA. Phylogenetic and sequence divergence analyses of our new data plus all previously published sequence data from those four loci collected in the USA, show that TcIV-USA is significantly different from any other previously defined T. cruzi DTUs. The significant level of genetic divergence between TcIV-USA and other T. cruzi DTUs should lead to an increased focus on understanding the epidemiological importance of this DTU, as well as its geographical range and pathogenicity in humans and domestic animals. Our findings further corroborate the fact that there is a high genetic diversity of the parasite in North America and emphasize the need for appropriate surveillance and vector control programs for Chagas disease in southern USA and Mexico.

Survey of Borreliae in ticks, canines, and white-tailed deer from Arkansas, U.S.A.

BACKGROUND: In the Eastern and Upper Midwestern regions of North America, Ixodes scapularis (L.) is the most abundant tick species encountered by humans and the primary vector of B. burgdorferi, whereas in the southeastern region Amblyomma americanum (Say) is the most abundant tick species encountered by humans but cannot transmit B. burgdorferi. Surveys of Borreliae in ticks have been conducted in the southeastern United States and often these surveys identify B. lonestari as the primary Borrelia species, surveys have not included Arkansas ticks, canines, or white-tailed deer and B. lonestari is not considered pathogenic. The objective of this study was to identify Borrelia species within Arkansas by screening ticks (n=2123), canines (n=173), and white-tailed deer (n=228) to determine the identity and locations of Borreliae endemic to Arkansas using PCR amplification of the flagellin (flaB) gene. METHODS: Field collected ticks from canines and from hunter-killed white-tailed were identified to species and life stage. After which, ticks and their hosts were screened for the presence of Borrelia using PCR to amplify the flaB gene. A subset of the positive samples was confirmed with bidirectional sequencing. RESULTS: In total 53 (21.2%) white-tailed deer, ten (6%) canines, and 583 (27.5%) Ixodid ticks (252 Ixodes scapularis, 161 A. americanum, 88 Rhipicephalus sanguineus, 50 Amblyomma maculatum, 19 Dermacentor variabilis, and 13 unidentified Amblyomma species) produced a Borrelia flaB amplicon. Of the positive ticks, 324 (22.7%) were collected from canines (151 A. americanum, 78 R. sanguineus, 43 I. scapularis, 26 A. maculatum, 18 D. variabilis, and 8 Amblyomma species) and 259 (37.2%) were collected from white-tailed deer (209 I. scapularis, 24 A. maculatum, 10 A. americanum, 10 R. sanguineus, 1 D. variabilis, and 5 Amblyomma species). None of the larvae were PCR positive. A majority of the flaB amplicons were homologous with B. lonestari sequences: 281 of the 296 sequenced ticks, 3 canines, and 27 deer. Only 22 deer, 7 canines, and 15 tick flaB amplicons (12 I. scapularis, 2 A. maculatum, and 1 Amblyomma species) were homologous with B. burgdorferi sequences. CONCLUSIONS: Data from this study identified multiple Borreliae genotypes in Arkansas ticks, canines and deer including B. burgdorferi and B. lonestari; however, B. lonestari was significantly more prevalent in the tick population than B. burgdorferi. Results from this study suggest that the majority of tick-borne diseases in Arkansas are not B. burgdorferi.

Wide dispersal and possible multiple origins of low-copy-number plasmids in rickettsia species associated with blood-feeding arthropods.

Plasmids are mobile genetic elements of bacteria that can impart important adaptive traits, such as increased virulence or antibiotic resistance. We report the existence of plasmids in Rickettsia (Rickettsiales; Rickettsiaceae) species, including Rickettsia akari, "Candidatus Rickettsia amblyommii," R. bellii, R. rhipicephali, and REIS, the rickettsial endosymbiont of Ixodes scapularis. All of the rickettsiae were isolated from humans or North and South American ticks. R. parkeri isolates from both continents did not possess plasmids. We have now demonstrated plasmids in nearly all Rickettsia species that we have surveyed from three continents, which represent three of the four major proposed phylogenetic groups associated with blood-feeding arthropods. Gel-based evidence consistent with the existence of multiple plasmids in some species was confirmed by cloning plasmids with very different sequences from each of two "Ca. Rickettsia amblyommii" isolates. Phylogenetic analysis of rickettsial ParA plasmid partitioning proteins indicated multiple parA gene origins and plasmid incompatibility groups, consistent with possible multiple plasmid origins. Phylogenetic analysis of potentially host-adaptive rickettsial small heat shock proteins showed that hsp2 genes were plasmid specific and that hsp1 genes, found only on plasmids of "Ca. Rickettsia amblyommii," R. felis, R. monacensis, and R. peacockii, were probably acquired independently of the hsp2 genes. Plasmid copy numbers in seven Rickettsia species ranged from 2.4 to 9.2 per chromosomal equivalent, as determined by real-time quantitative PCR. Plasmids may be of significance in rickettsial evolution and epidemiology by conferring genetic plasticity and host-adaptive traits via horizontal gene transfer that counteracts the reductive genome evolution typical of obligate intracellular bacteria.

Variations in orthodontic treatment planning decisions of Class II patients between virtual 3-dimensional models and traditional plaster study models.

INTRODUCTION: Study models provide invaluable information in treatment planning. Digital models have proved to be an effective measurement tool, but their use in treatment planning has not been studied. METHODS: Ten sets of records of Class II malocclusion subjects (dental study models, lateral cephalograms/tracings, panoramic radiographs, intraoral and extraoral photographs) were used for treatment planning by 20 orthodontists on 2 separate occasions. Digital models were used to evaluate the patients at 1 session and plaster models were used at the other session. Treatment recommendations were scored and compared for agreement. Eleven orthodontists served as the control group, looking at the records on 2 occasions with plaster models for agreement. RESULTS: Good agreement was noted for surgery (P = 1.00, kappa = 0.549), extractions (P = .360, kappa = 0.570), and auxiliary appliances (P = 1.00, kappa = 0.539) for the digital/plaster group. Agreement in the plaster/plaster group for surgery (P = 1.00, kappa = 0.671), extractions (P = 1.00, kappa = 0.626), and auxiliary appliances (P = .791, kappa = 0.672) was also good. Overall proportions of agreement ranged between 0.777 and 0.870 for digital/plaster and 0.818 and 0.873 for plaster/plaster. CONCLUSIONS: There was no statistical difference in intrarater treatment-planning agreement for Class II malocclusions based on the use of digital models in place of traditional plaster models. Digital orthodontic study models (e-models) are a valid alternative to traditional plaster study models in treatment planning for Class II malocclusion patients.

The accuracy of 4 panoramic units in the projection of mesiodistal tooth angulations.

The purpose of this study was to compare the mesiodistal tooth angulations determined with a typodont/skull testing device with the images of mesiodistal tooth angulations from 4 contemporary panoramic units (OP 100, Cranex 3+, Orthophos, PM 2002 EC). A typodont testing device was constructed, and the true mesiodistal tooth angulations relative to an orthodontic archwire were determined with a 3-dimensional coordinate-measuring machine and custom-designed software. A human skull served as the matrix into which the typodont was fixed for imaging. The skull was repeatedly imaged and repositioned 5 times for each panoramic unit. The images were scanned and digitized with custom software to determine the image mesiodistal angulations. Results revealed that the majority of image angles from the 4 panoramic units were statistically significantly different from the true angle measurements. However, definite trends were noted among the panoramic units. For the maxillary teeth, the images projected the anterior roots more mesially and the posterior roots more distally, creating the appearance of exaggerated root divergence between the canine and the first premolar. For the mandibular teeth, the images projected almost all roots more mesially than they really were, with the canine and the first premolar the most severely affected. The largest angular difference for adjacent teeth occurred between the mandibular lateral incisor and the canine, with relative root parallelism projected as root convergence. It was concluded that the clinical assessment of mesiodistal tooth angulation with panoramic radiography should be approached with extreme caution and with an understanding of the inherent image distortions.