Scrub typhus and the rickettsial diseases represent some of the oldest recognized vector-transmitted diseases, fraught with a rich historical aspect, particularly as applied to military/wartime situations. The vectors of Orientia tsutsugamushi were once thought to be confined to an area designated as the Tsutsugamushi Triangle. However, recent reports of scrub typhus caused by Orientia species other than O. tsutsugamushi well beyond the limits of the Tsutsugamushi Triangle have triggered concerns about the worldwide presence of scrub typhus. It is not known whether the vectors of O. tsutsugamushi will be the same for the new Orientia species, and this should be a consideration during outbreak/surveillance investigations. Additionally, concerns surrounding the antibiotic resistance of O. tsutsugamushi have led to considerations for the amendment of treatment protocols, and the need for enhanced public health awareness in both the civilian and medical professional communities. In this review, we discuss the history, outbreaks, antibiotic resistance, and burgeoning genomic advances associated with one of the world’s oldest recognized vector-borne pathogens, O. tsutsugamushi.

Abstract. Supercooling points and chill tolerance were compared among nymphs and adults of the ixodid ticks Dermacentor variabilis, Amblyomma americanum and Ixodes scapularis (Acari: Ixodidae).Supercooling points in the range of <‐22 to ‐18°C were observed for nymphs, and ‐22 to ‐8°C for adults.The lower lethal temperatures observed under dry conditions, ‐14 to ‐10°C, were warmer than the supercooling points, but still much colder than ‐4.8°C, the lowest temperature recorded from a likely tick habitat in southwestern Ohio.Based on our experiments, spontaneous freezing and direct chilling injury are not significant mortality factors in these species in the field.Mortality was observed between ‐5 and ‐3°C for A.americanum and D.variabilis nymphs chilled for 2 h while in direct contact with ice.This mortality is probably due to inoculative freezing.Given the requirement for a rather humid microhabitat for off‐host survival, these findings suggest that inoculative freezing is an important cause of overwintering mortality in these medically important species.

Recent studies indicated the predominance of Toxoplasma gondii haplogroup 12 in wildlife in USA. But still little is known of the genetic diversity of this parasite circulating in wildlife. In the present study, we tested coyotes (Canis latrans), red foxes (Vulpes vulpes), white-tailed deer (Odocoileus virginianus), and geese (Branta canadensis) from the state of Pennsylvania for T. gondii infection. Antibodies to T. gondii were found in 160 of 367 animals, including 92 (34.5%) of 266 coyotes, 49 (62.0%) of 79 white-tailed deer, 17 (85.0%) of 20 red fox, and two of two Canada geese tested by the modified agglutination test (cut off titer 1:25). Tissues from 105 seropositive animals were bioassayed in mice, and viable T. gondii was isolated from 29 animals, including 10 of 53 coyotes, 11 of 16 foxes, 7 of 49 deer, and one of one goose. DNA isolated from culture-derived tachyzoites of these isolates was characterized initially using multilocus PCR-RFLP markers. Nine genotypes were revealed, including ToxoDB PCR-RFLP #1 (4 isolates), #2 (2 isolates), #3 (4 isolates), #4 (6 isolates), #5 (4 isolates), #54 (1 isolate), #141 (1 isolate), #143 (1 isolate), and #216 (6 isolates), indicating high genetic diversity of T. gondii in wildlife in Pennsylvania. Pathogenicity of six T. gondii isolates (5 of #216 and #141) was determined in outbred Swiss Webster mice. Three of #216 and the #141 isolates were acute virulent to mice, and the other 2 #216 isolates were intermediate virulent. To determine the extent of genetic variation of these as well as a few recently reported virulent isolates from wildlife in North America, intron sequences were generated. Analysis of intron sequences and PCR-RFLP genotyping results indicated that the #216 isolates are likely derived from recombination of the clonal type I and III lineages. To determine if T. gondii virulence can be predicted by typing, we genotyped a collection of strains using PCR-RFLP markers for polymorphic genes ROP5, ROP16, ROP18 and GRA15, which are known to interact with host immune response. The results showed that there is an association of genotypes of ROP5 and ROP18 with mouse-virulence, however, additional gene(s) may also contribute to virulence in distinct T. gondii genotypes.

Nymphal ticks were consistently removed more successfully with commercial tools when compared with tweezers but with more difficulty than adults were removed. The commercial tick removal tools tested are functional for removal of nymphs and adults and should be considered as viable alternatives to medium-tipped tweezers.

Behavioral diapause was evaluated for overwintering Amblyomma americanum (L.) and Dermacentor variabilis (Say) laboratory strains. Ticks were placed in outdoor arenas on 30 Oct. 1996. On 9 Jan. 1997 specimens were removed for infestation on New Zealand white rabbits exposed to outside conditions or in a temperature and photoperiod controlled vivarium. Upon repletion, females were placed in tick rearing chambers indoors or retnmed to natural conditions. Both tick species readily attached when placed outside or in controlled conditions. Significantly more D. variabilis (63%) were attached than A. americanum (47%) three days post infestation. All females of both species failed to oviposit and perished when returned to natural winter conditions, while all successfully laid viable eggs under standard laboratory conditions. Based on the criteria of ability to attach, feed and oviposit, behavioral diapause was not in evidence during January for lone star and American dog tick females.