Ticks and Tick-Borne Diseases of Colorado, Including New State Records for Argas radiatus (Ixodida: Argasidae) and Ixodes brunneus (Ixodida: Ixodidae).

We report 28 species of ticks (Acari: Ixodida) from Colorado (CO). We include the soft ticks (Argasidae) Argas (Argas) cooleyi Kohls and Hoogstraal, Argas (Persicargas) radiatus Railliet, Carios (Alectorobius) concanensis (Cooley and Kohls), Carios (Alectorobius) kelleyi (Cooley and Kohls), Ornithodoros (Pavlovskyella) hermsi Wheeler et al., Ornithodoros (Pavlovskyella) parkeri Cooley, Ornithodoros (Pavlovskyella) turicata (Dugès), Otobius (Otobius) lagophilus Cooley and Kohls, and Otobius (Otobius) megnini (Dugès). We include the metastriate hard ticks (Ixodidae) Dermacentor (Americentor) albipictus (Packard), Dermacentor (Dermacentor) andersoni Stiles, Dermacentor (Dermacentor) parumapertus Neumann, Dermacentor (Dermacentor) variabilis (Say), Haemaphysalis (Aboimisalis) chordeilis (Packard), Haemaphysalis (Gonixodes) leporispalustris (Packard), and Rhipicephalus (Rhipicephalus) sanguineus Latreille. Prostriate hard ticks include Ixodes (Ixodiopsis) angustus Neumann, Ixodes (Phoeloioxdes) baergi Cooley and Kohls, Ixodes (Trichotoixodes) brunneus Koch, Ixodes (Scaphixodes) howelli Cooley and Kohls, Ixodes (Phoeloioxdes) kingi Bishopp, Ixodes (Phoeloioxdes) marmotae Cooley and Kohls, Ixodes (Ixodiopsis) ochotonae Gregson, Ixodes (Phoeloioxdes) sculptus Neumann, Ixodes (Ixodiopsis) soricis Gregson, Ixodes (Ixodes) spinipalpis Hadwen and Nuttall, Ixodes (Phoeloioxdes) texanus Banks, and Ixodes (Ixodiopsis) woodi Bishopp. Argas radiatus and Ixodes brunneus represent new state records. Review of collection reports revealed that inclusion of Ixodes (Multidentatus) auritulus (Neumann), Ixodes (Phoeloioxdes) cookei Packard, Ixodes (Phoeloioxdes) marxi Banks, and Ixodes (Ixodes) pacificus Cooley and Kohls is dubious or unconfirmed and, conversely, that C. concanensis and H. chordeilis have distributions that include CO. We list an additional five species occasionally detected and 13 exotic species intercepted in CO. Tick-host associations, geographical distributions, and medical/veterinary importance are included.

Acarine Hypopi (Sarcoptiformes: Acaridae) on Ixodes scapularis (Ixodida: Ixodidae) in Central North America.

Phoretic deutonymphs (hypopi) of Schwiebea sp. mites were detected during examinations of questing adult and nymphal blacklegged ticks, Ixodes scapularis Say, from vegetation at two woodland sites in northeastern Wisconsin, USA, during May and June, 2014. Collection sites were in Marinette and Oconto Counties, respectively. These hypopi are evidently the first reported from ticks in the Western Hemisphere, and they seem to be most similar morphologically to the putatively Holarctic species Schwiebea nova (Oudemans).

Equine piroplasmosis associated with Amblyomma cajennense Ticks, Texas, USA.

We report an outbreak of equine piroplasmosis in southern Texas, USA, in 2009. Infection prevalence reached 100% in some areas (292 infected horses). Amblyomma cajennense was the predominant tick and experimentally transmitted Theileria equi to an uninfected horse. We suggest that transmission by this tick species played a role in this outbreak.

Amblyomma triste (Acari: Ixodidae): new North American collection records, including the first from the United States.

New distribution records for the Neotropical tick, Amblyomma triste Koch, are identified from 27 specimens in 18 separate collections. These collections originated from six now recognized geographical foci in two states in the United States (Cochise and Santa Cruz Counties, Arizona, and Brewster and Jeff Davis Counties, Texas) and from import cattle, Bos taurus L., presented for entry at the United States border and originating in three Mexican states (Coahuila, Durango, and Sonora). For at least 67 yr, A. triste has existed in some areas of the United States as a cryptic species, and specimens there have been confused with and identified as Gulf Coast ticks, Amblyomma maculatum Koch, even by noted tick specialists. Most of the records reported in this study were from reidentified, archived specimens of putative A. maculatum.

Isolation of cell lines and a rickettsial endosymbiont from the soft tick Carios capensis (Acari: Argasidae: Ornithodorinae).

Soft ticks are medically important ectoparasites of birds and mammals that are found throughout the world. This report describes isolation and partial characterization of two embryonic cell lines, CCE2 and CCE3, from the seabird soft tick Carios capensis (Neumann). Sequencing of the mitochondrial 16S rRNA gene and karyology confirmed the lines were derived from C. capensis. CCE3 cells were diploid with a modal chromosome number of 20. The population doubling time for cell lines CCE2 and 3 in passage 40 was 6-9 d. A rickettsial endosymbiont, RCCE3, was co-isolated along with line CCE3. Nucleotide sequences of polymerase chain reaction (PCR) products generated using primers specific for rickettsial 17-kDa antigen, outer membrane protein (omp) A, ompB, and citrate synthase genes along with phylogenetic analyses demonstrated that RCCE3 is a previously uncultured endosymbiont. The rickettsia was identified as a symbiont of C. capensis, closely related to rickettsiae previously detected by PCR in C. capensis, Ornithodoros moubata (Murray) and Hemaphysalis sulcata Canestrini & Fanzago, a hard tick. RCCE3 caused a cytopathic effect in C. capensis host cells, and it was transferred to Ixodes scapularis Say cell line ISE6 for maintenance. The rickettsial endosymbiont was eliminated from CCE3 by treatment with oxytetracycline. Cell lines from C. capensis will be useful to researchers investigating interactions between soft ticks and microorganisms, soft tick physiology, and molecular biology. The rickettsia adds to the growing number of Rickettsia species that have been isolated in tick cell culture, and it is available for characterization.

Experimental transmission of West Nile virus (Flaviviridae: Flavivirus) by Carios capensis ticks from North America.

Seabird soft ticks, Carios capensis (Ixodida: Argasidae), originally collected from coastal Georgia, USA, were allowed to ingest a blood meal from pekin ducklings (Anas domesticus) infected with WNV. After 35 days of extrinsic incubation, the ticks transmitted virus to naive ducklings. WNV was detected via plaque assay and RTPCR in ticks and in tissues and serum of ducklings 7 days post infestation.

Identification of two phylogenetically related organisms from feces by PCR for detection of Salmonella spp.

Two previously reported PCR methods were evaluated to determine whether they are as sensitive and specific as conventional culture methods in detecting Salmonella spp. from feces. Bovine and equine feces were enriched overnight in brain heart infusion broth and assayed using PCR methods and primer sets described by other investigators. A total of 774 fecal specimens were tested using a primer set (invE-A primer set) that amplifies a region spanning the invasin E and A genes of Salmonella enterica serovar Typhimurium. A subset of these fecal specimens (306 of the 774 total) were tested using primers (hisJ primer set) that amplify a portion of the histidine transport J gene. The PCR required 24 h to obtain results, whereas it took 5 to 7 days to identify Salmonella spp. by culture. PCR detection of Salmonella spp. using the hisJ primers and the invE-A primers had a sensitivity of 93.3 and 80%, respectively, and a specificity of 85.6 and 98.6%, respectively, compared with bacterial culture. Amplification of 42 culture-negative fecal specimens (of 306 total specimens) generated a DNA fragment that corresponded to the molecular weight of the amplified hisJ gene. The hisJ-generated amplicons from six culture-negative and six culture-positive specimens were sequenced and analyzed using DNA sequence alignment and phylogenetic analysis software. A neighbor-joining dendrogram of the DNA sequences of both sets of hisJ amplicons revealed two distinct groups-one group of amplicons from culture-positive specimens identical to the hisJ gene of S. enterica serovar Typhimurium and a second group of amplicons from culture-negative specimens that were more closely related to hisJ of S. enterica serovar Typhimurium than to other hisJ sequences present in nucleotide databases.