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Spatial and temporal differences in the relative abundance of arthropod vectors are important factors that influence the risk of disease for mammalian hosts. Seasonal changes in the diversity and abundance of fleas (Siphonaptera) in Richardson's ground squirrel ( Urocitellus richardsonii ) burrows were studied at two sites in central Saskatchewan. A total of 225 fleas (151 at an urban site and 74 at a rural site) were collected. Flea prevalence differed among seasons at the urban site but not at the rural site. Of the nine flea species detected (eight at the urban site and six at the rural site), Oropsylla rupestris , O. bruneri , O. labis , O. tuberculata, and Aetheca wagneri are vectors of Yersinia pestis , the causative agent of plague. The presence and abundance of some fleas differed between sites and seasons. Neopsylla inopina and O. rupestris were the most abundant species at the urban site during the spring and summer, respectively, while O. bruneri was the most abundant species at the rural site. Our findings may have implications for the management of the black-tailed prairie dogs ( Cynomys ludovicianus ) in southwestern Saskatchewan because they coexist with U. richardsonii , are hosts for Oropsylla , and are at great risk of plague exposure/infection.

The geographical range of the American dog tick, Dermacentor variabilis (Say), in Canada continues to expand northwards into areas with colder winter temperatures. Understanding what influences the off-host survival of D. variabilis over winter is important for predicting the northern distributional limit of this species. A field experiment was conducted to determine the effect of snow cover on the off-host survival of unfed D. variabilis larvae from a population situated near the northern distributional limit in Saskatchewan, Canada. Ticks were placed in tubes within 1 of 9 field enclosures (tickaria) in early fall. The 9 tickaria were divided into 3 equal treatment groups: no snow cover, natural snow cover (maximum depth of 25 cm) and extra snow cover (maximum depth of 32 cm). Tick survival was measured in mid-winter and in early spring (i.e., ~17 and ~26 wk after start of experiment). The results showed that snow cover had a significant impact on the relative humidity, but not temperature, in the tickaria. Larvae in tickaria with no snow cover had a higher mortality rate (27%) than larvae in tickaria with natural snow cover (6%) or extra snow cover (4%). Although snow cover enhanced off-host survival, many individuals were able to survive in sub-zero temperatures under leaf litter in the absence of continuous snow cover for a period of 108 days. These findings have implications for the construction of predictive models to determine the northern distributional limits of D. variabilis, a species that is an important vector of several pathogens., (© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

A comparison was made of the supercooling points (SCPs) of questing Dermacentor variabilis adults from two populations located ca. 800 km apart on the Canadian prairies. This is also the first study to examine whether there are seasonal fluctuations in the SCP of questing D. variabilis adults. The SCPs of adult ticks from Lizard Lake Community Pasture, a recently established population in west-central Saskatchewan, varied over spring and summer, with the median SCP warming over time. In addition, the SCPs of ticks from Lizard Lake Community Pasture were significantly higher than those of adult ticks collected from Sandy Hook in Manitoba, a population that has been established for decades. The off-host survival of adults from Sandy Hook between summer and spring has been shown previously to be significantly greater than that of adults from Lizard Lake Community Pasture. The findings of the present study suggest that there may be geographical variation in the SCPs of D. variabilis adults which may be associated with differences in overwinter survival. The relatively low SCPs of questing D. variabilis adults, and the ability of some adults to survive off-host during winter, may be factors contributing to the range expansion of this tick species in Canada., (© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

The effects of temperature and relative humidity (RH) on female reproductive output, egg development and larval survival were determined for Rocky Mountain wood ticks (Dermacentor andersoni) from a prairie population (Chin Lakes, Alberta, Canada) near the northern distribution limit of this species. The responses of D. andersoni eggs and unfed larvae to different temperature (25 or 32 °C) and RH (35, 55, 75, 85 or 95%) regimes were compared to our previously published data (Diyes et al. 2021) for a northern prairie population of American dog ticks (Dermacentor variabilis). Oviposition by D. andersoni females took 21-30 days at 25 °C and 95% RH compared to 10-21 days for D. variabilis. The number of eggs laid by female ticks was strongly dependent on their engorgement weight, and D. andersoni females produced more eggs than D. variabilis females of an equivalent body weight. Eggs of D. andersoni took less time to develop at 32 °C than 25 °C with ≥ 85% RH, and hatched faster than those of D. variabilis. Larval survival times declined as temperature increased and RH decreased, but D. andersoni survived longer at 32 °C and ≤ 75% RH than D. variabilis. The interspecific differences in responses to the same temperature and humidity regimes indicate that D. andersoni is xerophilic, whereas D. variabilis is hydrophilic. Hence, 'prairie' populations of the Rocky Mountain wood tick occur in the drier grassland ecoregions but are absent in Aspen Parklands Ecoregion which is located to the north and east of the distributional range of D. andersoni., (© 2023. His Majesty in Right of Canada as represented by the Minister of Agriculture and Agri-Food and Chulantha P. Diyes, Neil B. Chilton.)

Dermacentor variabilis (Say) is expanding its range northwards in Saskatchewan and Manitoba (Canada). Knowledge of the relative ability of different life cycle stages to survive off-host over winter is important for assessing the potential for further range expansion and risk of pathogen transmission. In the present study, the off-host survival of D. variabilis adults was determined in outdoor terraria within a field plot close to Lizard Lake Community Pasture (LLCP) situated near the northern distributional limit in Saskatchewan. Only a small proportion (7%) of adult ticks in this relatively recently established population survived from mid-summer to late winter. Off-host survival had declined further to <4% by mid-spring. A significantly greater proportion of female ticks (6%) survived from mid-summer to mid-spring than did male ticks (1%). The supercooling points (SCPs), the temperature below 0 °C when body fluids spontaneously freeze, of the 17 ticks that survived through to mid-spring did not differ significantly from the SCPs of questing ticks in the previous summer, suggesting adult D. variabilis do not undergo cold hardening during winter diapause. The off-host survival of D. variabilis adults near LLCP was also much lower than that previously reported for adults from Sandy Hook in Manitoba, a population that has been established for decades. This finding suggests that there is geographical variation in the ability of D. variabilis in southern Canada to survive from mid-summer to the subsequent spring. However, the ecological and genetic factors influencing the survival of northern populations of D. variabilis requires further investigation., (© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

We detected the DNA of an Anaplasma bovis-like bacterium in blood specimens from 4 patients from the United States with suspected tickborne illnesses. Initial molecular characterization of this novel agent reveals identity to A. bovis-like bacteria detected in Dermacentor variabilis ticks collected from multiple US states.

The geographic distribution of the Rocky Mountain wood tick, Dermacentor andersoni Stiles, was determined in Alberta, Canada, by drag sampling at 86 and 89 sites during 2011 and 2012, respectively. Tick density and prevalence varied between years, averaging (range) 1.0 (0-26.2) and 5.9 (0-110) ticks/1,000 m2 in 2011 and 2012, respectively. Ticks were detected at 24.4% and 42.7% of the sites sampled in each respective year. Tick density and presence declined in a northerly direction to 51.6°N and in a westerly direction to ca. 113°W, except for a small area of high density at the edge of the Rocky Mountains in the southeastern portion of the province. Ticks were most abundant in the Dry Mixedgrass and Montane natural subregions and in areas with Brown Chernozemic, Regosol, and Solodized Solonetzic great soil groups. A logistic regression model indicated that tick presence was increased in the Dry Mixedgrass natural subregion and in regions with greater temperatures during the previous summer and normal winter precipitation but was reduced in areas with Dark Brown Chernozemic soils. The model will be useful for predicting tick presence and the associated risk of tick-borne diseases in the province., (© Her Majesty the Queen in Right of Canada, as represented by the Minister of Agriculture and Agri-Food Canada, 2021.)

Spotted fever group (SFG) rickettsiae are obligatory intracellular bacteria that cause disease in humans and other animals. Ixodid ticks are the principal vectors of SFG rickettsiae. The present study aimed to determine the prevalence and species identity of SFG rickettsiae in ticks and horses from urban and rural areas of western Cuba using PCR assays. Tick samples, collected from 79 horses, consisted of 14 Amblyomma mixtum adults, 111 Dermacentor nitens adults and 19 pools of D. nitens nymphs (2-5 individuals/pool). The PCR results revealed the presence of Rickettsia spp. in 64% of the A. mixtum adults, 16% of the D. nitens adults, and 11% of the pooled samples of D. nitens nymphs. In contrast, Rickettsia spp. was not detected in any of the 200 horse blood samples included in this study. DNA sequence data of the rickettsial 17 kDa antigen gene showed that Rickettsia amblyommatis was present in A. mixtum; and Rickettsia felis in D. nitens. This is the first report of R. felis in D. nitens in Cuba. The present study extends our knowledge of the potential vector spectrum and distribution of SFG rickettsiae pathogens in western Cuba., (© 2020 The Royal Entomological Society.)

The northern distributional limit of Dermacentor variabilis Say, the American dog tick, is expanding in Saskatchewan and Manitoba (western Canada). The ability of D. variabilis to continue to expand its range northwards will depend upon the ability of individuals within populations at the species distributional edge to withstand very low temperatures during winter. One component of cold hardiness is the supercooling point (SCP), the temperature below 0°C at which an individual freezes. In this study, the SCP was determined for 94 questing D. variabilis adults (44 females and 50 males) from an established population near Blackstrap Provincial Park in Saskatchewan. SCP values ranged from -18.2 to -6.7°C, with a median of -13.3°C. This suggests that host-seeking D. variabilis adults differ in their ability to survive exposure to subzero temperatures, for at least a short period of time, without freezing. The distribution of SCPs was bimodal, but there was no significant difference in SCP values between female and male ticks, and no relationship between SCP and tick body weight. It remains to be determined what factors contribute to the variation in SCP values among questing D. variabilis adults., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Psoroptes are a non-burrowing, ectoparasitic, mange-causing mite that has been documented in American bighorn sheep populations throughout the 19th and 20th centuries; however, it was not seen on Canadian bighorn sheep until 2006. The aim of this study was to determine the potential source of the Psoroptes outbreak in Canadian bighorn sheep. Morphological and molecular analyses were used to compare mites recovered from outbreak-associated bighorn sheep, pet rabbits in Canada, and on historically infested bighorn sheep in the USA. The results revealed that Psoroptes acquired from the Canadian and outbreak-associated American bighorn sheep were morphologically more similar to those collected from rabbits than mites on historically infested bighorn sheep. Outer opisthosomal setae lengths measured an average of 81.7 μm (±7.7 μm) in outbreak associated bighorn mites, 88.9 μm (±12.0 μm) in rabbit mites and 151.2 μm (±16.6 μm) in historically infested bighorn mites. The opisthosomal lobe morphology of bighorn mites in the outbreak herds was also more similar to that of rabbit mites, previously described as P. cuniculi , than historically infested bighorn mites, which match previous descriptions of P. ovis . This finding was supported by DNA sequence data of the mitochondrial cytochrome B gene. This is the first report of Psoroptes of the rabbit ecotype on bighorn sheep. The morphological and molecular data therefore support the hypothesis that the source of Psoroptes outbreak in Canadian bighorn sheep represented a disease spillover event from rabbits rather than transmission from infested American bighorn sheep populations., Competing Interests: None., (© 2021 The Authors.)

Female reproductive output and larval survival were determined for American dog ticks, Dermacentor variabilis (Say), from a recently established population near the northern distributional limit in Saskatchewan (Canada). Oviposition took 10-21 days at 25 °C and 95% relative humidity (RH). Temperature and relative humidity had a marked effect on egg development time and larval survival. Unfed larvae survived more than 100 days at 32 °C (with 95% RH) and 25 and 5 °C (with ≥ 85% RH). However, survival times declined markedly at lower relative humidities. In addition, 95% of the larvae placed in field enclosures survived for 140 days over winter during which they were exposed to sub-zero temperatures and 95-100% RH, while covered with snow. The median survival times (LT 50 ) of unfed larvae submerged underwater was 68 days. These results show that D. variabilis larvae in populations near the periphery of the northern distributional limit are adapted to cope with sub-zero temperatures in winter, and can survive in the temporary pools of water created by the spring snow melt.

Canine hepatozoonosis caused by Hepatozoon canis is a worldwide distributed tick-borne disease of domestic and wild canids that is transmitted by ingestion of Rhipicephalus sanguineus sensu lato (s.l.) ticks. The present study was aimed to determine the prevalence of Hepatozoon infections in 80 stray dogs from Havana Province in Cuba, and to confirm the species identity and phylogenetic relationships of the causative agent. Samples were screened by microscopical examination of thin blood smears for the presence of Hepatozoon spp. gamonts and by genus-specific SYBR green-based real-time PCR assay targeting the 18S rRNA gene. Direct microscopy examination revealed Hepatozoon gamonts in the peripheral blood of 8 dogs (10.0%; 95% CI: 4.80-18.0%), while 38 animals (47.5%; 95% CI: 36.8-58.4%) were PCR-positive, including all microscopically positive dogs. Hence, the agreement between the two detection methods was 'poor' (κ = 0.20). Hematological parameters did not differ significantly between PCR-positive and PCR-negative dogs (p > 0.05). The DNA sequences of the 18S rRNA gene of the Hepatozoon spp. from Cuban dogs showed a nucleotide identity >99% with those of 18S rRNA sequences of Hepatozoon canis isolates from Czech Republic, Brazil and Spain. Phylogenetic analysis revealed that obtained sequences clustered within the Hepatozoon canis clade, different from the Hepatozoon felis or Hepatozoon americanum clades. The present study represents the first molecular characterization of Hepatozoon canis in stray dogs within Cuba., (Copyright © 2020 Elsevier B.V. All rights reserved.)

It is often difficult to distinguish morphologically between closely related species of fleas (Siphonaptera). Morphological identification of fleas often requires microscopic examination of internal structures in specimens cleared using caustic solutions. This process degrades DNA and/or inhibits DNA extraction from specimens, which limits molecular-based studies on individual fleas and their microbiomes. Our objective was to distinguish between Oropsylla rupestris (Jordan), Oropsylla tuberculata (Baker), Oropsylla bruneri (Baker), and Oropsylla labis (Jordan & Rothschild) (Ceratophyllidae) using PCR-based single strand conformation polymorphism (SSCP) analyses and DNA sequencing. A 446 bp region of the nuclear 28S ribosomal RNA (rRNA) gene was used as the genetic marker. The results obtained for 36 reference specimens (i.e., fleas that were morphologically identified to species) revealed no intraspecific variation in DNA sequence, whereas the DNA sequences of the four species of Oropsylla differed from one another at two to six nucleotide positions. Each flea species also had a unique SSCP banding pattern. SSCP analyses were then used to identify another 84 fleas that had not been identified morphologically. DNA sequencing data confirmed the species identity of fleas subjected to SSCP. This demonstrates that PCR-SSCP combined with DNA sequencing of the 28S rRNA gene is a very effective approach for the delineation of four closely related species of flea., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Strepsiptera are an enigmatic order of insects with extreme sexual dimorphism which makes it difficult to "match-up" free-living adult males with parasitic conspecific females of the Stylopidia, and free-living females of the Mengenillidae using morphological characters. Species identification is further complicated for the Stylopidia because adult females are endoparasitic and neotenic. Therefore, we used DNA sequencing of the mitochondrial cytochrome c oxidase subunit 1 gene (cox1) to confirm the species identity of adult strepsipterans that were morphologically identified as Stylops advarians. These specimens, collected from Saskatoon (Saskatchewan, Canada), included one adult male, and eight females, the latter of which had been collected from solitary bees (Andrena milwaukeensis). Also included in the analyses were three pools of first-instar larvae that had emerged from three of the females. The results of the molecular analyses revealed that all specimens had an identical cox1 sequence, and belonged to a clade, with total statistical support (bootstrap value of 100%), that contained specimens of S. advarians from New York and Maine (USA). Hence, the results were consistent with the morphological identification of S. advarians. This study demonstrates the usefulness of a molecular approach for the identification of endoparasitic adult female and larval strepsipterans, life cycle stages that lack significant morphological characters for species identification.

Total genomic (g)DNA from 100 American dog ticks (Dermacentor variabilis) collected from humans, dogs, raccoons, and skunks near Minnedosa (Manitoba, Canada) in 2005 was tested for the presence of Moellerella wisconsensis (Gammaproteobacteria: Enterobacteriales) using PCR. Although two gDNA samples derived from ticks attached to two striped skunks (Mephitis mephitis) contained M. wisconsensis DNA, it is unlikely that D. variabilis is a vector of this bacterium. Genomic DNA prepared from the washes of the external surfaces of these two ticks (i.e., before DNA extraction from the whole tick) and another two ticks attached to same skunks were also PCR positive for M. wisconsensis. This suggests that ticks acquired the bacterium by physical contact with contaminated or infected skunks. However, it does not exclude the possibility that the ticks may have also imbibed the bacterium from their host blood and lymph. Nonetheless, the results of this molecular study suggest that the four adult D. variabilis represent biological indicators of the presence of M. wisconsensis in association with their vertebrate hosts (i.e., striped skunks). Additional work is needed to determine if M. wisconsensis is present in the blood and lymph of striped skunks in southwestern Manitoba and if there are potential health risks for persons coming into contact with infected animals., (© Wildlife Disease Association 2020.)

Background: In a warmer and more globally connected Arctic, vector-borne pathogens of zoonotic importance may be increasing in prevalence in native wildlife. Recently, Bartonella henselae, the causative agent of cat scratch fever, was detected in blood collected from arctic foxes (Vulpes lagopus) that were captured and released in the large goose colony at Karrak Lake, Nunavut, Canada. This bacterium is generally associated with cats and cat fleas, which are absent from Arctic ecosystems. Arctic foxes in this region feed extensively on migratory geese, their eggs, and their goslings. Thus, we hypothesized that a nest flea, Ceratophyllus vagabundus vagabundus (Boheman, 1865), may serve as a vector for transmission of Bartonella spp., Methods: We determined the prevalence of Bartonella spp. in (i) nest fleas collected from 5 arctic fox dens and (ii) 37 surrounding goose nests, (iii) fleas collected from 20 geese harvested during arrival at the nesting grounds and (iv) blood clots from 57 adult live-captured arctic foxes. A subsample of fleas were identified morphologically as C. v. vagabundus. Remaining fleas were pooled for each nest, den, or host. DNA was extracted from flea pools and blood clots and analyzed with conventional and real-time polymerase chain reactions targeting the 16S-23S rRNA intergenic transcribed spacer region., Results: Bartonella henselae was identified in 43% of pooled flea samples from nests and 40% of pooled flea samples from fox dens. Bartonella vinsonii berkhoffii was identified in 30% of pooled flea samples collected from 20 geese. Both B. vinsonii berkhoffii (n = 2) and B. rochalimae (n = 1) were identified in the blood of foxes., Conclusions: We confirm that B. henselae, B. vinsonii berkhoffii and B. rochalimae circulate in the Karrak Lake ecosystem and that nest fleas contain B. vinsonii and B. henselae DNA, suggesting that this flea may serve as a potential vector for transmission among Arctic wildlife.

PCR-based single-strand conformation polymorphism (SSCP) analyses combined with DNA sequencing of the prokaryotic 16S ribosomal (r) RNA gene encompassing the hypervariable V4 region was used to determine the bacterial composition of Rocky Mountain wood ticks (Dermacentor andersoni) attached to Richardson's ground squirrels (Urocitellus richardsonii) and questing on vegetation in southern Saskatchewan, Canada. The bacteria present in questing adult ticks from Saskatchewan Landing Provincial Park included Rickettsia peacockii, a Francisella-like endosymbiont (FLE) and an Arsenophonus-like endosymbiont. Bacteria in the adult and nymphal ticks attached to U. richardsonii collected from Beechy included R. peacockii, a FLE, and several other genera (e.g., Ralstonia, Sphingobium, Comamonas and Pseudomonas). The bacteria detected in D. andersoni in the present study are consistent with the findings of other studies that have characterized the microbiome of this tick species in the USA using next generation sequencing. This result demonstrates that the SSCP-based approach used in this study is cost- and time-effective for examining bacterial composition in ticks.

Passive and active surveillance for the blacklegged tick, Ixodes scapularis Say, in the Canadian province of Saskatchewan was conducted over a 9-yr period (2009-2017). More than 26,000 ixodid ticks, representing 10 species, were submitted through passive surveillance. Most (97%) of these were the American dog tick, Dermacentor variabilis (Say). Of the 65 I. scapularis adults submitted, 75% were collected from dogs. Infection rates of Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti in I. scapularis were 12%, 8%, and 0%, respectively. Although the I. scapularis submitted by passive surveillance were collected from five of seven ecoregions in central and southern Saskatchewan, they were most frequent in the Moist Mixed Grassland and Aspen Parklands. In contrast, no I. scapularis were collected from the extensive field sampling conducted at multiple sites in different ecoregions across the province. Hence, there is no evidence of I. scapularis having established a breeding population in Saskatchewan. Nonetheless, continued surveillance for blacklegged ticks is warranted given their important role as a vector of medically and veterinary important pathogens, and because they have recently become established across much of the southern portions of the neighboring province of Manitoba., (© Her Majesty the Queen in Right of Canada, as represented by the Minister of Agriculture and Agri-Food Canada, 2019.)

The genetic variation and taxonomic status of the four morphologically-defined species of Macropostrongyloides in Australian macropodid and vombatid marsupials were examined using sequence data of the ITS+ region (=first and second internal transcribed spacers, and the 5.8S rRNA gene) of the nuclear ribosomal DNA. The results of the phylogenetic analyses revealed that Ma. baylisi was a species complex consisting of four genetically distinct groups, some of which are host-specific. In addition, Ma. lasiorhini in the common wombat (Vombatus ursinus) did not form a monophyletic clade with Ma. lasiorhini from the southern hairy-nosed wombat (Lasiorhinus latifrons), suggesting the possibility of cryptic (genetically distinct but morphologically similar) species. There was also some genetic divergence between Ma. dissimilis in swamp wallabies (Wallabia bicolor) from different geographical regions. In contrast, there was no genetic divergence among specimens of Ma. yamagutii across its broad geographical range or between host species (i.e. Macropus fuliginosus and M. giganteus). Macropostrongyloides dissimilis represented the sister taxon to Ma. baylisi, Ma. yamagutii and Ma. lasiorhini. Further morphological and molecular studies are required to assess the species complex of Ma. baylisi.

The phylogenetic relationships of 42 species of cloacinine nematodes belonging to three tribes (Coronostrongylinea, Macropostrongylinea and Zoniolaiminea) were examined based on sequence data of the first and second internal transcribed spacers (ITS-1 and ITS-2) of the nuclear ribosomal DNA. All nematodes examined are parasites of Australian macropodid marsupials. None of the three nematode tribes was monophyletic. Paraphyly was also encountered in three genera: Papillostrongylus, Monilonema and Wallabinema. Species within the genus Thallostonema were limited to a single host genus (i.e. Thylogale), whereas species within the five principal genera (Coronostrongylus, Macropostrongylus, Popovastrongylus, Wallabinema and Zoniolaimus) were found to occur in multiple host genera. Potential modes of evolution among these nematodes are discussed.

Human granulocytic anaplasmosis (HGA) is an emerging disease in Canada because of range expansion by the arthropod vector, Ixodes scapularis. These ticks carry the Ap-ha variant of Anaplasma phagocytophilum (Ap-ha), which has been implicated in causing HGA, and the Ap-variant 1, which is not associated with human infection. We report the detection of 13 genotypes of the ankyrin (ankA) gene among 76 infected blacklegged ticks. Haplotype network and phylogenetic analyses revealed that the ankA genotypes corresponding to the Ap-ha variant did not form a monophyletic assemblage. They formed two distinct clades (Clades I and III), one of which was genetically more similar in nucleotide and amino acid sequences to genotypes of Ap-variant 1 that comprised Clade II. Additional work is needed to explore the evolutionary history of A. phagocytophilum in North America, and to determine if there are differences in pathogenicity or clinical symptoms associated with the two divergent groups of the Ap-ha variant given the significant differences in ankA amino acid sequence., (Crown Copyright © 2018. Published by Elsevier GmbH. All rights reserved.)

PCR and DNA sequencing were used to determine the prevalence of Anaplasma bovis in Rocky Mountain wood ticks (Dermacentor andersoni) collected in Alberta and Saskatchewan, Canada. These analyses revealed that A. bovis DNA was present in 35 (2.1%) of 1679 ticks collected at 12 of the 24 localities. The discovery of A. bovis in host-seeking female and male D. andersoni from multiple locations in southern Canada may have important implications for diagnosis of anaplasmosis in Canada., (Crown Copyright © 2018. Published by Elsevier GmbH. All rights reserved.)

Sequences of the first and second internal transcribed spacers (ITS1 + ITS2) of nuclear ribosomal DNA were employed to determine whether the congeneric assemblages of species of the strongyloid nematode genus Cloacina, found in the forestomachs of individual species of kangaroos and wallabies (Marsupialia: Macropodidae), considered to represent species flocks, were monophyletic. Nematode assemblages examined in the black-striped wallaby, Macropus (Notamacropus) dorsalis, the wallaroos, Macropus (Osphranter) antilopinus/robustus, rock wallabies, Petrogale spp., the quokka, Setonix brachyurus, and the swamp wallaby, Wallabia bicolor, were not monophyletic and appeared to have arisen by host colonization. However, a number of instances of within-host speciation were detected, suggesting that a variety of methods of speciation have contributed to the evolution of the complex assemblages of species present in this genus.

Pharyngostrongylus thylogale n. sp. (Nematoda: Strongylida) is described from the stomach of the red-legged pademelon, Thylogale stigmatica (Gould) (Marsupialia: Macropodidae) from north-eastern Queensland and Papua New Guinea, having formerly been confused with P. iota Johnston & Mawson, 1939. Pharyngostrongylus thylogale n. sp. differs from all congeners in having 12 labial crown elements rather than eight or 16. Pharyngostrongylus iota was found in T. stigmatica, but only in southern Queensland and northern New South Wales, in the subspecies T. s. wilcoxi, compared with P. thylogale n. sp. which was found in T. s. stigmatica in northern Queensland and T. s. oriomo in Papua New Guinea. Differences in the sequences of the first and second internal transcribed spacers (ITS-1 and ITS-2) of the nuclear ribosomal DNA of P. thylogale n. sp. and ten congeners support the erection of the new species, and the validity of the morphospecies examined. However, results of the phylogenetic analyses of the molecular data also provide evidence for the existence of cryptic species within P. kappa Mawson, 1965. No obvious co-evolutionary relationships were observed between parasite species and their macropodid marsupial hosts.

Sequences of the internal transcribed spacers of nuclear ribosomal DNA (ITS-1 and ITS-2) were determined for species of the genus Rugopharynx and Rugonema labiatum, nematodes from the stomachs of macropodid marsupials. Phylogenetic analyses of the aligned sequence data were conducted. The relationships provided molecular support for all species currently recognised, some of which are based on minor morphological differences and on multilocus enzyme electrophoretic data, but also indicated that additional, cryptic species exist within the genus. In addition, the genus Rugonema is placed as a synonym of Rugopharynx, its sole species becoming Rugopharynx labiatum n. comb. The molecular data provided some insights into the evolution of complex buccal capsule morphologies within the genus, but there was no evidence of co-evolution between the macropodid hosts and their parasites.

A phylogeny for seven species of Cyclostrongylus and the monotypic genus Spirostrongylus (Nematoda: Chabertiidae), all highly host specific parasites of the oesophagi of wallabies (Marsupialia: Macropodidae), was constructed using sequence data for the first and second internal transcribed spacers (ITS-1 and ITS-2) of the nuclear ribosomal DNA. There was no evidence for co-speciation, or for the sympatric or synxenic speciation of Cyclostrongylus alatus and Cyclostrongylus perplexus, both of which are parasites of Macropus rufogriseus. Rather, host switching, correlating with geographical distributions, appeared to provide some explanation of the pattern of speciation observed., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Schistosomiasis is a neglected tropical disease that affects more than 200 million people worldwide. The main disease-causing agents, Schistosoma japonicum, S. mansoni and S. haematobium, are blood flukes that have complex life cycles involving a snail intermediate host. In Asia, S. japonicum causes hepatointestinal disease (schistosomiasis japonica) and is challenging to control due to a broad distribution of its snail hosts and range of animal reservoir hosts. In China, extensive efforts have been underway to control this parasite, but genetic variability in S. japonicum populations could represent an obstacle to eliminating schistosomiasis japonica. Although a draft genome sequence is available for S. japonicum, there has been no previous study of molecular variation in this parasite on a genome-wide scale. In this study, we conducted the first deep genomic exploration of seven S. japonicum populations from mainland China, constructed phylogenies using mitochondrial and nuclear genomic data sets, and established considerable variation between some of the populations in genes inferred to be linked to key cellular processes and/or pathogen-host interactions. Based on the findings from this study, we propose that verifying intraspecific conservation in vaccine or drug target candidates is an important first step toward developing effective vaccines and chemotherapies against schistosomiasis.

The phylogenetic relationships of the endemic (or largely endemic) Australasian trichostrongylin nematode families Herpetostrongylidae, Mackerrastrongylidae and Nicollinidae as well as endemic trichostrongylin nematodes currently placed in the families Trichostrongylidae and Molineidae were examined using the complete large subunit (28S) ribosomal RNA gene. The Herpetostrongylinae proved to be monophyletic. However, representatives of the Nicollinidae nested with the Herpetostrongylinae. The Mackerrastrongylidae was also a monophyletic group and included Peramelistrongylus, currently classified within the Trichostrongylidae. The Globocephaloidinae, currently considered to be a subfamily of the Herpetostrongylidae, was excluded from the family in the current analysis. Ollulanus and Libyostrongylus, included for the first time in a molecular phylogenetic analysis, were placed within the Trichostrongylidae. This study provided strong support for the Herpetostrongylidae (including within it the Nicollinidae, but excluding the Globocephaloidinae) and the Mackerrastrongylidae as monophyletic assemblages. Additional studies are required to resolve the relationships of the remaining endemic Australasian trichostrongylin genera.

To investigate the agreement between available serologic tests for the detection of antibodies against Anaplasma phagocytophilum and Borrelia burgdorferi, 50 serum samples from horses of unknown clinical status and at low risk for infection were tested. In addition to a point-of-care enzyme-linked immunosorbent assay (pocELISA), the evaluated tests included 2 indirect fluorescent antibody tests (IFATs) for antibodies against A. phagocytophilum and an IFAT, an ELISA confirmed with Western blot, and the Lyme multiplex assay for antibodies against B. burgdorferi. For each pair-wise comparison between serologic tests, the difference in the proportion of seropositive results as well as kappa and the prevalence-adjusted, bias-adjusted kappa were calculated. The proportion of seropositive results differed significantly in each pairwise comparison of tests for detection of antibodies against A. phagocytophilum, and between the pocELISA and IFAT as well as between the pocELISA and Lyme multiplex assay for detection of antibodies against B. burgdorferi. Agreement based on kappa varied from poor to fair while agreement was improved when evaluating prevalence-adjusted, bias-adjusted kappa. Lack of agreement may be explained by differences in methodology between the evaluated tests, cross-reactivity or false-positive and false-negative tests. In addition to the limitations of serologic test interpretation in the absence of clinical disease, this data suggest that screening of horses for exposure to tick-borne diseases in nonendemic areas may not be warranted., (© 2015 The Author(s).)

Equine granulocytic anaplasmosis (EGA) and Lyme borreliosis (LB) are an emerging concern in Canada. We estimated the seroprevalence of EGA and equine LB by testing 376 convenience serum samples from 3 provinces using a point-of-care SNAP(®) 4Dx(®) ELISA (IDEXX Laboratories, Westbrook, Maine, USA), and investigated the agreement between the point-of-care ELISA and laboratory-based serologic tests. The estimated seroprevalence for EGA was 0.53% overall (0.49% in Saskatchewan, 0.71% in Manitoba), while the estimated seroprevalence for LB was 1.6% overall (0.49% in Saskatchewan, 2.86% in Manitoba). There was limited agreement between the point-of-care ELISA and an indirect fluorescent antibody test for EGA (kappa 0.1, PABAK 0.47) and an ELISA/Western blot combination for LB (kappa 0.23, PABAK 0.71). While the SNAP(®) 4Dx(®) ELISA yielded expected seroprevalence estimates, further evaluation of serologic tests for the purposes of disease exposure recognition may be needed.

The DNA sequences of the mitochondrial (mt) 12S rRNA and tRNA(Val) genes were characterized for 82 blacklegged ticks (Ixodes scapularis) that were genetically identical for Domains IV and V of the mt 16S rRNA gene. Thirty-one haplotypes, differed in sequence by 1-9 bp, were detected among the 82 ticks. Most nucleotide alterations in DNA sequence did not affect the stability of the secondary structures of the RNAs. The magnitude of the DNA sequence variation in the mt 12S rRNA and tRNA(Val) genes among blacklegged ticks suggests that this region of the mitochondrial genome has potential as a genetic marker for examining the population genetics and phylogeography of I. scapularis., (Copyright © 2015. Published by Elsevier Ltd.)

Passive surveillance of ticks on horses in Saskatchewan revealed that the horses were parasitized by 3 species, Dermacentor albipictus, D. andersoni, and D. variabilis. The nymphs and adults of D. albipictus occurred on horses earlier in the year than did adults of the 2 other species.

The complete DNA sequences and secondary structure of the mitochondrial (mt) 16S ribosomal (r) RNA gene were determined for six Ixodes scapularis adults. There were 44 variable nucleotide positions in the 1252 bp sequence alignment. Most (95%) nucleotide alterations did not affect the integrity of the secondary structure of the gene because they either occurred at unpaired positions or represented compensatory changes that maintained the base pairing in helices. A large proportion (75%) of the intraspecific variation in DNA sequence occurred within Domains I, II and VI of the 16S gene. Therefore, several regions within this gene may be highly informative for studies of the population genetics and phylogeography of I. scapularis, a major vector of pathogens of humans and domestic animals in North America., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

This study examined genetic variation within and among species of Cloacina found in the swamp wallaby (Wallabia bicolor) collected at different localities along the eastern coast of Australia, and evaluated geographical distance as a potential driver for genetic variation. The first and second internal transcribed spacers (ITS-1 and ITS-2, respectively) of the nuclear ribosomal DNA were used to characterize individuals of 11 morphospecies of Cloacina that parasitize W. bicolor. The results of the molecular analyses revealed multiple genotypes for the nine morphospecies of Cloacina (i.e. Cloacina annulata, Cloacina edwardsi, Cloacina eos, Cloacina gallardi, Cloacina mawsonae, Cloacina papillata, Cloacina papillatissima, Cloacina pollux, and Cloacina wallabiae) for which multiple individuals were available for analysis. However, phylogenetic analyses of the sequence data revealed that for each morphospecies, there was no subdivision of individuals into distinct clades based on geographical region from which they were collected. Additional studies are needed to determine the drivers of genetic variation in cloacinid nematodes, and hence increase our understanding of the diversity of parasitic nematodes in macropodid marsupials., (Copyright © 2014 Elsevier B.V. All rights reserved.)

We developed PCR-based assays to distinguish a human pathogenic strain of Anaplasma phagocytophilum, Ap-ha, from Ap-variant 1, a strain not associated with human infection. The assays were validated on A. phagocytophilum-infected black-legged ticks (Ixodes scapularis) collected in Canada. The relative prevalence of these 2 strains in I. scapularis ticks differed among geographic regions.