Conventional PCR provides Leishmania species characterization with even a small amount of biological material. Species-specific primers have been a widely used alternative; however, nonspecific amplifications are a reality, interfering with PCR efficiency. In endemic areas with multiple etiological agents for leishmaniasis, there is a requirement for higher specificity of primers. This study evaluates 3 pairs of primers described for the identification and characterization of Leishmania infantum. Primers RV1/RV2, LEISH1/LEISH2, and FLC2/RLC2 were used with the DNA of L. infantum, Leishmania amazonensis, and Leishmania braziliensis. An initial temperature curve was performed (52–62 C) to determine the optimal annealing temperature, followed by a dilution curve of Leishmania DNA (500 pg/µl, 50 pg/µl, 5 pg/µl, 500 fg/µl, 50 fg/µl, 5 fg/µl, and 0.5 fg/µl) to be used for analytical sensitivity. RV1/RV2 PCR amplified L. infantum and L. amazonensis at all analyzed temperatures; LEISH1/LEISH2 PCR amplified all 3 species of Leishmania, although at some temperatures L. infantum was specifically amplified, and, finally, FLC2/RLC2 PCR amplified only L. infantum at all temperatures analyzed. In terms of sensitivity, RV1/RV2 PCR detected 1 fg of L. infantum DNA and 100 pg of L. amazonensis DNA; LEISH1/LEISH2 PCR detected 1 fg of L. infantum DNA, 100 fg of L. amazonensis DNA, and 10 fg of L. braziliensis DNA; and FLC2/ RLC2 PCR detected 10 fg of L. infantum DNA. Thus, PCR with FLC2/RLC2 primers is best suited for the molecular characterization of L. infantum, especially in areas where there is an incidence of more than 1 Leishmania species, such as South America.

Recent studies of louse ectoparasites from mummies have developed robust data sets that allow a true epidemiological approach to the prehistory of louse parasitism. One epidemiological principle is that the binomial of overdispersion is normally negative, meaning that in a host population, parasites are aggregated in a few individuals. We demonstrate the overdispersion of lice in 3 different prehistoric communities that differ along 3 axes or variables: environmental setting, socioeconomic status, and cultural affiliation. Distinct cultural practices could have been involved in different patterns of louse infestation. Prevalence, intensity, and abundance of infestations exhibit statistically significant differences between the communities. We also find differences in prevalence between subadults and adults that contrasted by cultural affiliation and suggest conditions different from those seen today. We show that overall prevalence was affected primarily by ecological setting, not socioeconomic status nor cultural affiliation. These findings demonstrate that statistical analysis of archaeological data can reveal the states of infestation in past populations with lifestyles not seen in modern people. Our approach paves the way for future comparisons of subpopulations within archaeological communities.

The northern cottonmouth, Agkistrodon piscivorus (Lacépède, 1789) (Viperidae: Crotalinae), occurs in the southeastern United States and is one of the few semiaquatic vipers in the world. Three proteocephalid tapeworms of the genus Ophiotaenia La Rue, 1911 (Cestoda), have been described from this venomous snake. A critical evaluation of type specimens and tapeworms recently collected from A. piscivorus has revealed that only 2 species, Ophiotaenia marenzelleri (Barrois, 1898) and Ophiotaenia grandis La Rue, 1911 (new synonym Ophiotaenia agkistrodontis [Harwood, 1933]), are specific parasites of this crotaline viper; both species are redescribed here. Ophiotaenia grandis was originally described from a mixture of 2 species: ‘true’ O. grandis, which is indistinguishable from O. agkistrodontis described 22 yr later and thus considered to be a junior synonym of O. grandis, and O. marenzelleri. Ophiotaenia marenzelleri, also reported from the pygmy rattlesnake, Sistrurus miliarius (Linnaeus, 1766), is a larger tapeworm with a massive scolex, a large cirrus sac, a very large, annular vaginal sphincter near the equatorial gonopore, and an oval, 3-layered embryophore surrounded by a nearly square hyaline outer membrane. Ophiotaenia grandis is much smaller and more slender and has a narrower scolex, a smaller cirrus sac and vaginal sphincter, a 2-layered embryophore, and a distinctly pre-equatorial gonopore. In addition to O. marenzelleri and O. grandis, other Ophiotaenia species typical of semiaquatic snakes (Colubridae: Natricinae) may be present in the northern cottonmouth, which serves only as a postcyclic or occasional host. There is also a tabular summary of 18 species of Ophiotaenia from semiaquatic snakes worldwide, with information on their hosts, distribution, and taxonomically important characters, including key measurements.

Yak (Bos grunniens) farming is an important part of Mongolia's livestock industry. Yaks survive in harsh mountain environments; provide meat, milk, and wool; and serve as a mode of transportation. In Mongolia, yaks are frequently raised alongside other livestock animals such as cattle, Bactrian camels, sheep, goats, and horses. Recently, we demonstrated that Babesia bovis, Babesia bigemina, and Babesia naoakii—parasites with the potential to cause clinical bovine babesiosis—infect not only cattle but also Bactrian camels in Mongolia. However, yaks have never been surveyed for Babesia infections in this country. In the present study, we surveyed yaks in 8 Mongolian provinces: Bayankhongor, Bayan-Ulgii, Khovd, Khovsgol, Omnogovi, Ovorkhangai, Uvs, and Zavkhan. Blood samples were taken and deoxyribonucleic acid (DNA) was extracted from 375 yaks. Furthermore, Giemsa-stained thin smears were prepared from 315 of the 375 blood samples and then examined for the microscopic detection of Babesia parasites. Microscopy revealed that 34 (10.8%) of 315 blood smears were positive for Babesia parasites. All 375 DNA samples were then tested for B. bovis, B. bigemina, and B. naoakii infection using specific polymerase chain reaction assays. We observed that 238 (63.5%) yaks in all surveyed provinces and 8 (2.1%) yaks in 3 provinces (Bayankhongor, Bayan-Ulgii, and Omnogovi) were positive for B. bovis and B. bigemina, respectively. However, all yaks tested were negative for B. naoakii. This epidemiological survey, the first to report Babesia infection in Mongolian yaks, suggests that disease management strategies for yaks in this country should further address bovine babesiosis.

Bot flies (Oestridae: Cuterebrinae) are obligate mammalian parasites that complete the larval stage of their life cycle under the skin of their host. Most detailed studies of bot fly larval disease ecology have been conducted in temperate deciduous zone rodent systems. To understand the relative importance of seasonal and spatial factors, as well as factors intrinsic to the host, in underpinning the likelihood and extent of parasitism by bot flies in non-rodent hosts as well as in arid-land ecosystems, we examined the dynamic for black-tailed jackrabbit (Lepus californicus) and desert cottontail (Sylvilagus audubonii) parasitism by bot fly larvae (Cuterebra spp.) across 7 repeatedly sampled sites spread across approximately 500 km of the Chihuahuan Desert ecoregion of southern New Mexico. This environment is characterized by a climate that includes hot dry summers and cool to cold dry winters, as well as strongly seasonal summer monsoonal rains. Lagomorphs are a common mid-sized mammal in these landscapes. Bot fly parasitism was strongly seasonal, with peak prevalence and abundance in the spring, and there was spatial variation in the extent of parasitism between collection sites. Additionally, jackrabbits in better body condition were less likely to be parasitized (as indicated by kidney fat index). We did not find sex-based differences in bot fly parasitism between male and female jackrabbits. Thus, in arid-land ecoregions, abiotic factors are likely the primary driver of the bot fly–host interaction, whereas factors intrinsic to the host were of secondary importance for characterizing the interactions of bot flies and lagomorphs.

Dicyemid mesozoans usually consist of 10 to 40 cells. They are characterized by 2 distinct embryos, vermiform and infusoriform, that develop within the axial cell of the adult. The means of escape of each embryo from the parent body was studied in Dicyema japonicum and Dicyema misakiense, parasites of Octopus sinensis. There were no differences in means of escape between species or embryo type, apparently due to morphological constraints whereby the parents (nematogen or rhombogen) share a similar body organization. Escapes were effected through the gap between adjacent peripheral cells of the adult, rupturing the axial cell membrane and the membrane that envelopes the embryo. After the embryo escaped, the path was closed by the enveloping membrane left behind by the embryo. Vermiform embryos can escape from any region of the body, although more embryos were observed to escape from anterior regions than from posterior regions. Infusoriform embryos escaped from both anterior and posterior regions in the axial cell, with more embryos observed to escape from the posterior regions. The different escape regions for the 2 types of embryo are presumably related to the adult body plan lacking a genital opening, so each different type of embryo has its appropriate site of escape.

Areas of endemism are the smallest units in biogeography and can be defined as biologically unique areas comprising taxa with common geographic limits to their distributions. High beta diversity within Amazonia is often related to turnover among these areas. For decades, evolutionary biologists have tried to comprehend the mechanisms generating and maintaining the spatial structure and high diversity of free-living Amazonian organisms, particularly birds. However, few studies have tried to analyze these patterns among their parasites. Host and parasite associations involve shared history that may allow us to better understand the fine-scale evolutionary history of the host. Here we compare the coevolutionary patterns among 2 avian host species with distinct patterns of genetic structure in northern Amazonia, Dendrocincla fuliginosa (Aves: Dendrocolaptidae) and Dixiphia pipra (Aves: Pipridae), and their ectoparasitic lice (Insecta: Phthiraptera), Furnaricola sp. ex Dendrocincla fuliginosa, Myrsidea sp. ex Dixiphia pipra, and Tyranniphilopterus sp. ex Dixiphia pipra. We obtained sequences of the mitochondrial gene cytochrome oxidase subunit I from hosts and parasites collected on opposite banks of the Negro and Japurá rivers, which delimit 3 areas of endemism in northern Amazonia: Napo, Jau, and Guiana. Our results demonstrate that the Negro River is a geographical barrier for both Furnaricola sp. and its avian host, Dendrocincla fuliginosa. Phylogenies of both hosts, Dendrocincla fuliginosa, and the parasites, Furnaricola sp., show monophyletic clades on opposite margins of the river that are not sister taxa. These clades have a mean uncorrected p-distance of 17.8% for Furnaricola sp. and 6.0% for Dendrocincla fuliginosa. Thus, these parasite clades constitute distinct evolutionary lineages and may even be distinct species. In contrast, Dixiphia pipra has no population structure associated with either river. Accordingly, data from their lice Myrsidea sp. indicate weak support for different clades on opposite margins of the Negro River, whereas data from their lice Tyranniphilopterus sp. indicate weak structure across the Japurá. This study is a first step toward understanding the effects of biogeographic history on permanent ectoparasites and suggests that host biogeographic history is to some extent a determinant of the parasite's history. Furthermore, the parasite's evolutionary history is an additional source of information about their hosts' evolution in this highly diverse region of northern Amazonia.

The chigger species Acomatacarus coahuilensis n. sp. (Acariformes: Trombidiformes: Leeuwenhoekiidae) inhabiting the neck scales of the wary spiny lizard Sceloporus cautus Smith, 1938 (Squamata: Iguania: Phrynosomatidae), from Chihuahuan Desert Province, Coahuila, Mexico, is described herein. The new mite species is assigned to the genus Acomatacarus Ewing, 1942, based on a palptarsus with 6 branched setae plus the tarsala (ω). This new species can be separated from the other species in this genus mainly based on the combination of the following characters: presence of mastisetae (MTa) on tarsus III and tibiala (φ) on tibia III, subterminala (ζ₁), parasubterminala (z), and pretarsala (ζ₂) on tarsus I, 2 genualae (σ_1-2) on genu I, onychotriches and claw-like empodium of legs I–III, and absence of mastisetae on tibia III. In addition to the new species, the gastrointestinal nematode Skrjabinoptera phrynosoma (Ortlepp, 1922) Schulz, 1927 (Physalopteridae), was also found, which represents a new host record in the wary spiny lizard. This study reports the third species of the genus Acomatacarus recorded from Mexico and the first record of chigger mites and helminth parasites associated with Sc. cautus.

Cytauxzoon felis is a tick-transmitted intraerythrocytic apicomplexan infecting felids in the southeastern and midwestern United States. Bobcats (Lynx rufus) are the natural wildlife reservoir of C. felis, where in enzootic areas prevalence can reach 100%. Domestic cats can be subclinically infected with C. felis or can develop cytauxzoonosis. Two studies have documented the presence of C. felis in domestic cats in Illinois; these studies have shown a limited number of cases submitted to specialty labs. During 2014–2018, we obtained blood samples collected by veterinary staff from 514 cats that were apparently healthy and 74 cats that were suspected of cytauxzoonosis. These samples were screened using a sensitive, nested PCR to detect the presence of C. felis DNA. We herein document frequent occurrences of cytauxzoonosis (8–15 cases/year from 4 veterinary clinics) and 12.5% prevalence of subclinical infections in southern Illinois, a locality showing a sharp increase in cases of cytauxzoonosis. Our results suggest a high risk of cytauxzoonosis in southern Illinois, despite only recently being recognized in the area. We found no specific risk factors for cytauxzoonosis or subclinical infections in this location. In addition, cases of cytauxzoonosis occur every month of the year (with the highest frequency in summer) and therefore tick prevention should be used in domestic cats in enzootic regions throughout the year.

Bovine anaplasmosis, caused by Anaplasma marginale, is widespread in cattle in the southeast United States. The pathogen is biologically transmitted by Dermacentor spp. ticks, and mechanically transmitted by biting flies and via fomites. Despite high reported regional seroprevalence, Dermacentor spp. are rare on cattle in the southeast. To identify other putative An. marginale vectors, and to characterize cattle exposure to other tick-borne pathogens, we collected ticks from Arkansas cattle herds in 2020–2022. Recognizing that deer are important hosts for some of the same tick species that parasitize cattle, we also collected ticks from hunter-killed deer in the fall and winter of 2021. Ticks were screened for bacteria in the family Anaplasmataceae using qPCR. Positive samples were further amplified using a PCR assay targeting the groEL gene, and the resulting amplicons were sequenced for identification. A total of 3,794 ticks were collected, the majority of which were Amblyomma americanum. Amblyomma americanum was the most common species on cattle, and Ixodes scapularis was most common on deer. No ticks were positive for An. marginale, though Anaplasma phagocytophilum was detected in deer-collected I. scapularis, as well as in a single engorged Am. americanum from cattle. Amblyomma americanum from cattle were infected with Ehrlichia ewingii, Ehrilichia chaffeensis, and Panola Mountain Ehrlichia. Cattle in Arkansas are exposed to several ehrlichial pathogens and may also be exposed to An. phagocytophilum. The importance of these pathogens, particularly Panola Mountain Ehrlichia, in causing cattle disease in Arkansas deserves further study, as does the importance of mechanical transmission of An. marginale in bovine anaplasmosis epidemiology.