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A survey was conducted in the lowland areas of Binh Dinh and Phu Yen provinces in south central Vietnam to study the diversity of digenean cercariae within freshwater snails. These regions are known as highly endemic areas for the transmission of liver flukes, Fasciola spp. and Opisthorchis viverrini. In total, 13 snail species were found, and 3 of these were dominant, comprising 51.9% of all collected snails, i.e., Bithynia funiculata, Melanoides tuberculata, and Radix auriculata. Cercariae were found in 5 snail species, and among these, Indoplanorbis exustus had the highest overall prevalence of infection (38.5%). No cercarial infections were found in viviparid and ampullariid snails. Morphological identification resulted in 10 cercariae types that were illustrated and measured. Among them, xiphidiocercariae and echinostomata were the most common, causing 70.55% of all cercariae infections. Bithynia species had the most diverse digenean fauna. Molecular data revealed the presence of cercariae of 13 digenean species, including O. viverrini. The 18S rDNA sequences of echinostomata and xiphidiocercariae type 3, which were shed from different snail hosts, were similar to 2 and 3 digenean species, respectively. In conclusion, our findings highlight the diversity of digenean fauna in Binh Dinh and Phu Yen provinces.

The sucking louse fauna associated with Mongolian mammals is inadequately known. We provide a list of 25 species of sucking lice recorded from Mongolian rodents including previously published records, and new records of specimens collected during an expedition to northwestern Mongolia in 2015. Hoplopleura inagakii Ono and Hasegawa and Polyplax cricetulis Chin are newly recorded from Mongolia and 2 new host associations in Mongolia are recorded for Hoplopleura acanthopus (Burmeister). We describe Hoplopleura altaiensis n. sp., from the Gobi Altai mountain vole, Alticola barakshin Bannikov (type host) with an additional specimen from Alticola strelzowi (Kastchenko) (Strelzow's mountain vole). Both sexes of the new species are illustrated with scanning electron micrographs and line drawings. We note small morphological differences in the shape of the female subgenital plate between specimens prepared for scanning electron microscopy versus those prepared for light microscopy following DNA extraction.

Anenterotrema is a small genus of dicrocoeliids (Digenea: Dicrocoeliidae) containing 6 species found in Neotropical bats. Members of this genus are characterized by the lack of digestive system organs and, unlike the majority of dicrocoeliids, parasitize the intestine of their definitive hosts. In this study, we have morphologically examined newly collected specimens belonging to four species of Anenterotrema from Brazil, Ecuador, and Panama. Based on the data in original descriptions and our analysis of quality new specimens, we resurrected Anenterotrema freitasi, previously synonymized with A. eduardocaballeroi. We also described a new species of Anenterotrema from Molossus molossus in the Brazilian Amazon. The new species differs from congeners in several morphological features, most prominently in the size and structure of its highly muscular, protuberant ventral sucker. It is also characterized by the lack of the semi-circular thickening of the tegument around the oral sucker typical for some members of the genus. We used partial DNA sequences of the large ribosomal subunit gene (28S) and mitochondrial cytochrome c oxidase subunit 1 gene (cox1) to test the monophyly of Anenterotrema, and study the interrelationships and determine the inter- and intraspecific variation of three Anenterotrema spp. collected from different bat species in Brazil, Ecuador and Panama.

White grub metacercariae were found in the livers and kidneys of diseased specimens of an introduced channel catfish, Ictalurus punctatus (Rafinesque), in Vietnam. Based on morphological features and 28S rDNA sequence analysis, the isolated metacercariae were identified as Dollfustrema bagarii (Digenea: Bucephalidae) MoravecSey. Histopathological examination shows that encysted metacercariae can change the tissue structure of the infected organs and is often accompanied by haemorrhaging and the presence of eosinophilic granular cell infiltration. Degenerative changes were also observed.

Grouse and ptarmigan (Galliformes) harbor fairly diverse helminth faunas that can impact the host's health, including filarial nematodes in the genus Splendidofilaria. As host and parasite distributions are predicted to shift in response to recent climate change, novel parasites may be introduced into a region and impose additional stressors on bird populations. Limited information is available on the prevalence of filariasis in Alaska galliforms. To date, no molecular surveys have been completed. Past studies relied on examining blood smears or total body necropsies, which are time-consuming and may not detect filarial parasites with low prevalence in hosts. Therefore, we developed a TaqMan probe-based real-time PCR assay targeting the cytochrome c oxidase 1 gene (COI) of Splendidofilaria to decrease processing times and increase sensitivity as well as provide baseline data on the diversity of filariid infections in galliform species in Alaska. We screened a combined total of 708 galliform samples (678 unique individual birds) from different tissues (blood, muscle, and lung) for the presence of filarial DNA across the state of Alaska. Real-time PCR screening revealed an overall prevalence of filarial infection of 9.5% across species: Bonasa umbellus (0%, n = 23), Dendragapus fuliginosus (0%, n = 8), Falcipennis canadensis (26.8%, n = 198), Lagopus lagopus (2.6%, n = 274), Lagopus leucura (0%, n = 23), Lagopus muta (3%, n = 166), and Tympanuchus phasianellus (12.5%, n = 16). We observed microfilarial infections throughout most of Alaska except in Arctic regions and the Aleutian Islands where viable vectors may not be present.

In total, 341 fleas belonging to 16 species were collected from 78 host mammals belonging to 10 species in Panamá from 2010 to 2016. The cat flea, Ctenocephalides felis (Bouché) predominated on domestic dogs and was also recorded from domestic cats, the raccoon, Procyon lotor (Linnaeus) and the common opossum, Didelphis marsupialis Linnaeus. The largest number of flea species (7) was recorded from D. marsupialis and the most common flea on that host was the ctenophthalmid, Adoratopsylla intermedia copha Jordan. One Oriental rat flea, Xenopsylla cheopis (Rothschild), was collected from D. marsupialis. Native rodents were parasitized by indigenous ceratophyllid, rhopalopsyllid, and stephanocircid fleas. The Mexican deermouse, Peromyscus mexicanus (Saussure), was parasitized by six species of ceratophyllids belonging to the mostly Central American genera, Baculomeris, Jellisonia, Kohlsia and Plusaetis. The long-tailed singing mouse, Scotinomys xerampelinus (Bangs), was parasitized by Plocopsylla scotinomi Tipton and Méndez, the only species of stephanocircid flea known from Central America. Twenty-six pools of extracted flea DNA representing 5 flea species (C. felis, Pulex echidnophagoides (Wagner), Pulex simulans Baker, A. intermedia copha, and P. scotinomi) and 79 individual fleas were all real-time polymerase chain reaction negative for Rickettsia felis, Rickettsia typhi, and Bartonella henselae.

Natural history collections (NHCs) are important resources for a diverse array of scientific fields. Recent digitization initiatives have broadened the user base of NHCs, and new technological innovations are using materials generated from collections to address novel scientific questions. Simultaneously, NHCs are increasingly imperiled by reductions in funding and resources. Ensuring that NHCs continue to serve as a valuable resource for future generations will require the scientific community to increase their contribution to and acknowledgement of collections. We provide recommendations and guidelines for scientists to support NHCs, focusing particularly on new users that may be unfamiliar with collections. We hope that this perspective will motivate debate on the future of NHCs and the role of the scientific community in maintaining and improving biological collections.

In Vietnam, helminthioses remain a major threat to public health and contribute to the maintenance of poverty in highly endemic regions. Through increased awareness of the damaging effects caused by helminthioses, the Vietnamese government has implemented many national programs over the past 30 years for the prevention and control of the most important helminthioses, such as, lymphatic filariasis, soil transmitted helminths, food borne zoonotic helminths, and others. Various control strategies have been applied to reduce or eliminate these worms, e.g. mass drug administration, economic development, control of vectors or intermediate hosts, public health interventions through education, proper composting procedures for excreta potentially containing helminth eggs, and the expansion of food supply chains and improved technologies for the production and inspection of food products. These control measures have resulted in a significant reduction in the distribution and transmission of helminth infections and have improved the overall living conditions and health outcomes of the Vietnamese citizens. However, the persistence of several helminth diseases continues in some endemic areas, especially where poverty is widespread and local traditions include the consumption of raw foods, especially fish and meats. This manuscript provides an overview of the helminth infection prevention and control programs conducted in Vietnam, their achieved results, learned lessons, and future works.

PURPOSE: We aimed to characterize the urinary microbiome of catheterizing patients with neurogenic lower urinary tract dysfunction (NLUTD) and to evaluate differences based on type of bladder management or frequency of urinary tract infections. MATERIALS AND METHODS: This is a prospective, cross-sectional study of urine samples from asymptomatic, catheterizing patients with (NLUTD) who used either clean intermittent catheterization (CIC) or indwelling catheters. Patients without symptoms of urinary tract infection provided a catheterized urine sample for urinalysis, culture, and bacterial community microbiome analysis. RESULTS: Ninety-five patients submitted urine for analysis, of these 69 had sufficient sequence reads (>1203) for microbiome analysis. Patients with low bacterial signal amplification were associated with use of vaginal estrogen, no intradetrusor botulinum toxin A (BTX-A) use, and no growth on standard urine culture. The most abundant operational taxonomic units (OTU) were from the phylum Proteobacteria, classified as Enterobacteriaceae and Escherichia. Alpha diversity varied among those who used indwelling catheters versus CIC, and those who underwent BTX-A injection versus not. On linear discriminate analysis, the relative abundance of the OTU identified as Pseudomonas was higher among patients using indwelling catheters relative to CIC. The OTU identified as Aerococcus was at a higher relative abundance among males compared to females. CONCLUSIONS: Enterobacteriaceae and Escherichia were the most abundant genera in the urinary microbiome of patients with NLUTD. Urinary microbiome diversity varied based on bladder management type. Future clinical correlations between microbiome of neurogenic patients and clinical presentation may help guide treatment strategies.

Crassiphialinae Sudarikov, 1960 is a large subfamily of the Diplostomidae Poirier, 1886 with a complex taxonomic history. It includes a diversity of species parasitic in the intestines of avian and mammalian definitive hosts worldwide. Posthodiplostomum Dubois, 1936 is a large and broadly distributed crassiphialine genus notorious for its association with diseases in their fish second intermediate hosts. In this study, we generated partial 28S rDNA and cytochrome c oxidase subunit 1 (cox1) mtDNA gene sequences of digeneans beloning to seven crassiphialine genera. The 28S sequences were used to study the interrelationships among crassiphialines and their placement among other major diplostomoidean lineages. Our molecular phylogenetic analysis and review of morphology does not support subfamilies currently recognized in the Diplostomidae; therefore, we abandon the current subfamily system of the Diplostomidae. Molecular phylogenetic analyses suggest the synonymy of Posthodiplostomum, Ornithodiplostomum Dubois, 1936 and Mesoophorodiplostomum Dubois, 1936 ; morphological study of our well-fixed adult specimens and review of literature revealed lack of consistent differences among the three genera. Thus, we synonymize Ornithodiplostomum and Mesoophorodiplostomum with Posthodiplostomum. Our phylogenetic analyses suggest an Old World origin of Posthodiplostomum followed by multiple dispersal events among biogeographic realms. Furthermore, our analyses indicate that the ancestors of these digeneans likely parasitized ardeid definitive hosts. Four new species of Posthodiplostomum collected from birds in the New World as well as one new species of Posthodiplostomoides Williams, 1969 from Uganda are described.

The systematic position of Urotrema Braun, 1900 and the family Urotrematidae Poche, 1926 have always been controversial. Due to its unusual morphological characteristics, lack of knowledge of the life cycle or details of its excretory system, this family was placed within different higher taxonomic groups of digeneans. Despite being one of the most enigmatic digenean families in terms of its phylogenetic affinities, DNA sequence data for Urotrematidae were lacking. Here, we evaluate the phylogenetic relationships of Urotrema using newly obtained partial sequences of the 28S rRNA gene from Urotrema specimens collected in North, Central and South America including the type species U. scabridum Braun, 1900, as well as previously published sequences of digeneans. Our study has demonstrated that Urotrema is phylogenetically closest (100% branch support) to members of Parabascus Looss, 1907 belonging to the family Pleurogenidae Looss, 1899. Thus, the family Urotrematidae becomes a junior synonym of the Pleurogenidae. Urotrema forms a 100% supported clade among the Pleurogenidae, parasitic in warm-blooded vertebrates. However, the phylogenetic relationships and exact systematic position of the remaining 3 genera currently placed in the Urotrematidae remains unclear and requires additional studies as their allocation is mostly based on the terminal posterior position of the genital pore and cirrus-sac. According to our results the genus Parabascus appears to be paraphyletic and requires further detailed phylogenetic and morphological analyses.

Museum specimens play an increasingly important role in predicting the outcomes and revealing the consequences of anthropogenically driven disruption of the biosphere. As ecological communities respond to ongoing environmental change, host–parasite interactions are also altered. This shifting landscape of host–parasite associations creates opportunities for colonization of different hosts and emergence of new pathogens, with implications for wildlife conservation and management, public health, and other societal concerns. Integrated archives that document and preserve mammal specimens along with their communities of associated parasites and ancillary data provide a powerful resource for investigating, anticipating, and mitigating the epidemiological, ecological, and evolutionary impacts of environmental perturbation. Mammalogists who collect and archive mammal specimens have a unique opportunity to expand the scope and impact of their field work by collecting the parasites that are associated with their study organisms. We encourage mammalogists to embrace an integrated and holistic sampling paradigm and advocate for this to become standard practice for museum-based collecting. To this end, we provide a detailed, field-tested protocol to give mammalogists the tools to collect and preserve host and parasite materials that are of high quality and suitable for a range of potential downstream analyses (e.g., genetic, morphological). Finally, we also encourage increased global cooperation across taxonomic disciplines to build an integrated series of baselines and snapshots of the changing biosphere. Los especímenes de museo desempeñan un papel cada vez más importante tanto en la descripción de los resultados de la alteración antropogénica de la biosfera como en la predicción de sus consecuencias. Dado que las comunidades ecológicas responden al cambio ambiental, también se alteran las interacciones hospedador-parásito. Este panorama cambiante de asociaciones hospedador-parásito crea oportunidades para la colonización de diferentes hospedadores y para la aparición de nuevos patógenos, con implicancias en la conservación y manejo de la vida silvestre, la salud pública y otras preocupaciones de importancia para la sociedad. Archivos integrados que documentan y preservan especímenes de mamíferos junto con sus comunidades de parásitos y datos asociados, proporcionan un fuerte recurso para investigar, anticipar y mitigar los impactos epidemiológicos, ecológicos y evolutivos de las perturbaciones ambientales. Los mastozoólogos que recolectan y archivan muestras de mamíferos, tienen una oportunidad única de ampliar el alcance e impacto de su trabajo de campo mediante la recolección de los parásitos que están asociados con los organismos que estudian. Alentamos a los mastozoólogos a adoptar un paradigma de muestreo integrado y holístico y abogamos para que esto se convierta en una práctica estándarizada de la obtención de muestras para museos. Con este objetivo, proporcionamos un protocolo detallado y probado en el campo para brindar a los mastozoólogos las herramientas para recolectar y preservar materiales de parásitos y hospedadores de alta calidad y adecuados para una gran variedad de análisis subsecuentes (e.g., genéticos, morfológicos, etc.). Finalmente, también abogamos por una mayor cooperación global entre las diversas disciplinas taxonómicas para construir una serie integrada de líneas de base y registros actuales de nuestra cambiante biosfera.

The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) pandemic reveals a major gap in global biosecurity infrastructure: a lack of publicly available biological samples representative across space, time, and taxonomic diversity. The shortfall, in this case for vertebrates, prevents accurate and rapid identification and monitoring of emerging pathogens and their reservoir host(s) and precludes extended investigation of ecological, evolutionary, and environmental associations that lead to human infection or spillover. Natural history museum biorepositories form the backbone of a critically needed, decentralized, global network for zoonotic pathogen surveillance, yet this infrastructure remains marginally developed, underutilized, underfunded, and disconnected from public health initiatives. Proactive detection and mitigation for emerging infectious diseases (EIDs) requires expanded biodiversity infrastructure and training (particularly in biodiverse and lower income countries) and new communication pipelines that connect biorepositories and biomedical communities. To this end, we highlight a novel adaptation of Project ECHO’s virtual community of practice model: Museums and Emerging Pathogens in the Americas (MEPA). MEPA is a virtual network aimed at fostering communication, coordination, and collaborative problem-solving among pathogen researchers, public health officials, and biorepositories in the Americas. MEPA now acts as a model of effective international, interdisciplinary collaboration that can and should be replicated in other biodiversity hotspots. We encourage deposition of wildlife specimens and associated data with public biorepositories, regardless of original collection purpose, and urge biorepositories to embrace new specimen sources, types, and uses to maximize strategic growth and utility for EID research. Taxonomically, geographically, and temporally deep biorepository archives serve as the foundation of a proactive and increasingly predictive approach to zoonotic spillover, risk assessment, and threat mitigation.

Skrjabinoptera vietnamensis n. sp. is described from specimens recovered from the stomach of Eutropis macularia in north-central Vietnam. The new species is characterized by the medium-sized male worms (6.7–8.7 mm in length and 154–182 µm in width) relative to known members of the genus, 2 pointed spicules of unequal length (87–112 µm and 56–72 µm in length), and 10 pairs of caudal papillae. Female worms are larger than male worms (10.7–18.4 mm in length and 264–411 µm in width), with the vulva situated in the anterior part, and embryonated, elliptical eggs, 35–46 µm long by 20–24 µm wide. Skrjabinoptera vietnamensis n. sp. represents the ninth species assigned to the genus and the first species recorded from the Oriental region. Partial sequences of the 18S ribosomal RNA gene (rDNA), and cytochrome c oxidase subunit 1 (COI) are provided for the new species. The molecular phylogenetic position of the genus Skrjabinoptera is briefly discussed.

The chigger, Euschoengastia pipistrelli Brennan, is a trombiculid mite that infests a variety of vespertilionid bats in North America. It has been reported from at least 9 species of bats from 18 U.S. states. However, nothing is available on the actual in situ infestation and ultrastructure of this chigger. Here we document some stereoscopic photographs of the infestation as well as a scanning electron micrograph of the mite from a common bat species. We also provide a summation of host and state records for this chigger.

Falcaustra vietnamensis n. sp. is described from the small intestine of Physignathus cocincinus from north-central Vietnam. The new species is characterized by the large male worms (20.2-28.8 mm in length and 557-724 μm in width) relative to known members of the genus, 2 sharply pointed alate spicules of equal length (1,128-1,256 μm in length), gubernaculum including 2 separate pieces, 1 ventral with a pointed distal end and 1 dorsal with a blunt distal end (164-192 μm and 155-172 μm in length, respectively), and 12 pairs of caudal papillae. Female worms are larger than male worms (24.2-34.1 mm in length and 532-735 μm in width), with the vulva situated in the posterior half of body, and elliptical eggs, 60-70 μm long by 42-47 μm wide. Falcaustra vietnamensis n. sp. represents the 38th species assigned to the genus and the third species recorded from a lizard host in the Oriental biogeographical region. Partial sequences of the 18S ribosomal RNA gene (rDNA), internal transcribed spacer regions (ITS), and cytochrome c oxidase subunit 1 (COI) are provided for the new species. The molecular phylogenetic position of the genus Falcaustra is briefly discussed.

Anenterotrema is a small genus of dicrocoeliids (Digenea: Dicrocoeliidae) containing 6 species found in Neotropical bats. Members of this genus are characterized by the lack of digestive system organs and, unlike the majority of dicrocoeliids, parasitize the intestine of their definitive hosts. In this study, we have morphologically examined newly collected specimens belonging to four species of Anenterotrema from Brazil, Ecuador, and Panama. Based on the data in original descriptions and our analysis of quality new specimens, we resurrected Anenterotrema freitasi, previously synonymized with A. eduardocaballeroi. We also described a new species of Anenterotrema from Molossus molossus in the Brazilian Amazon. The new species differs from congeners in several morphological features, most prominently in the size and structure of its highly muscular, protuberant ventral sucker. It is also characterized by the lack of the semi-circular thickening of the tegument around the oral sucker typical for some members of the genus. We used partial DNA sequences of the large ribosomal subunit gene (28S) and mitochondrial cytochrome c oxidase subunit 1 gene (cox1) to test the monophyly of Anenterotrema, and study the interrelationships and determine the inter- and intraspecific variation of three Anenterotrema spp. collected from different bat species in Brazil, Ecuador and Panama.

Documenting emergence of invasive species in new areas is vital to understanding spatiotemporal patterns of invasions, propagule pressure, and the risk of establishment. Salvator merianae (Argentine Black and White Tegu) has established multiple unconnected populations in southern and central Florida, and additional sightings have been reported elsewhere in the state. In 2018, land managers in Georgia received >20 reports of this species in the wild. To evaluate the probability of establishment, we assembled verified records of the non-native Argentine Black and White Tegu in Georgia over the past 9 years. We report on 47 tegu observations throughout Georgia, with a concentration of sightings (n = 38) in Toombs and Tattnall counties. In 2019, we used modified Havahart® traps and captured adult male and female tegus at 1 of our 3 locations during 3085 corrected trap nights. While we did not find evidence of a well-established population (i.e., varied size structure of tegus captured) with our limited trapping effort, we suspect Argentine Black and White Tegus are breeding in Toombs and Tattnall counties due to the concentration of captures and reports of adult males and females, the consistent reports of adults across years, the confirmed presence of the species in 2018, 2019, and 2020, and the reproductive capacity (i.e., turgid testes and secondary follicles) of individuals captured. Ongoing introductions of tegus from captivity are likely to maintain high propagule pressure in the southeastern United States. Effective early detection, funded rapid response networks, and public outreach to solicit reports of sightings of Argentine Black and White Tegus are critical to prevent establishment and associated ecological impacts of this invasive species elsewhere in the southeastern US.

Two new species of sucking lice (Phthiraptera: Anoplura), Hoplopleura granti n. sp. (Hoplopleuridae) and Polyplax megacephalus n. sp. (Polyplacidae), are described from Grant's rock mouse, Micaelamys granti (Wroughton), from Northern Cape Province, South Africa. Adults of both sexes are illustrated via line drawings and differential interference contrast microscopy images, and are compared with previously described related species that parasitize related hosts: Hoplopleura patersoni Johnson from Aethomys chrysophilus (de Winton) (red veld rat), Aethomys kaiseri (Noack) (Kaiser's aethomys), and Micaelamys namaquensis (A. Smith) (Namaqua rock mouse); Hoplopleura aethomydis Kleynhans from M. namaquensis; Polyplax praomydis Bedford from A. chrysophilus and M. namaquensis; and Polyplax solivaga Johnson from A. chrysophilus. It is not known if these new species of lice are vectors of any pathogens.

A total of 52 specimens of the Pacific seabream Acanthopagrus pacificus Iwatsuki, Kume et Yoshino from the Gulf of Tonkin off Vietnam were examined for monogeneans. Twenty fish were parasitised by 101 individuals of five monogenean species, including two known species Allodiscocotyla diacanthi Unnithan, 1962 and Heterapta chorinemi (Tripathi, 1956), as well as three new species, Polylabroides tienyenensis sp. n., Polylabroides tonkinensis sp. n. and Metacamopia lebedevi sp. n. Polylabroides tienyenensis and P. tonkinensis are morphologically more similar to Polylabroides guangdongensis Zhang et Yang, 2001 in comparison with other species within the genus, based on the absence of small spines on the cirrus. However, P. tonkinensis is distinguished from P. guangdongensis by fewer clamps on the haptor and by the different shapes of the large spines on the cirrus. Similarly, P. tienyenensis differs from P. guangdongensis by vaginal ducts with fewer branches, fewer clamps and smaller egg size. Metacamopia lebedevi is distinguished from Metacamopia chorinemi (Yamaguti, 1953) by the arrangement of testes (one row vs two rows), diverticula absent in the oesophagus, and the number of anchor pairs (one vs two); it differs from Metacamopia oligoplites Takemoto, Amato et Luque, 1996 by the smaller haptor, shape and absence of small sclerotised hooks, the number of ribs in their clamps, and the position of the testes; it can be separated from Metacamopia indica (Unnithan, 1962) by having fewer testes and lacking sclerotised structures in the vagina. The present study also provides the measurements for A. diacanthi, H. chorinemi, and proposes a new key to all species of Polylabroides.

Members of the sucking louse genus Pedicinus are ectoparasites of cercopithecid primates in Africa, Asia, and Gibraltar. Pedicinus gabonensis n. sp. is described based on adult male and female specimens collected from the mandrill (Mandrillus sphinx) in Gabon. The new species is compared morphologically with other members of the genus Pedicinus, and a nuclear elongation factor 1 alpha gene sequence is provided. Host associations and geographical distributions of the 18 previously recognized species of the genus, and of Pedicinus gabonensis n. sp., are reviewed. Updated identification keys are provided for males and females of all known valid species of Pedicinus.

In their Perspective “Rigorous wildlife disease surveillance” (10 July, p. 145), M. Watsa et al. underscore the value of One Health approaches to stimulate integration across currently siloed efforts in zoonotic research and mitigation. To achieve comprehensive decentralized pathogen surveillance, there is an urgent need to develop environmental and biodiversity infrastructure in biodiverse countries experiencing high rates of habitat conversion, wildlife trafficking, and human-wildlife interactions.

Natural history collections spanning multiple decades provide fundamental historical baselines to measure and understand changing biodiversity. New technologies such as next generation DNA sequencing have considerably increased the potential of museum specimens to address significant questions regarding the impact of environmental changes on host and parasite/pathogen dynamics. We developed a new technique to identify intestinal helminth parasites and applied it to shrews (Eulipotyphla: Soricidae) because they are ubiquitous, occupy diverse habitats, and host a diverse and abundant parasite fauna. Notably, we included museum specimens preserved in various ways to explore the efficacy of using metabarcoding analyses that may enable identification of helminth symbiont communities from historical archives. We successfully sequenced the parasite communities (using 12S mtDNA, 16S mtDNA, 28S rDNA) of 23 whole gastrointestinal tracts. All gastrointestinal tracts were obtained from the Museum of Southwestern Biology, USA, and from recent field collections, varying both in time since fixation (ranging from 4 months to 16 years) and preservation method (70% or 95% ethanol stored at room temperature, or flash frozen in liquid nitrogen and stored at −80 °C). Our proof of concept demonstrates the feasibility of applying next generation DNA sequencing techniques to authoritatively identify the parasite/pathogen communities within whole gastrointestinal tracts from museum specimens of varying age and fixation, and the value of future preservation of host-associated whole gastrointestinal tracts in public research archives. This powerful approach facilitates future comparative examinations of the distributions and interactions among multiple associated groups of organisms through time and space.

The Beringian Coevolution Project (BCP), a field program underway in the high northern latitudes since 1999, has focused on building key scientific infrastructure for integrated specimen-based studies on mammals and their associated parasites. BCP has contributed new insights across temporal and spatial scales into how ancient climate and environmental change have shaped faunas, emphasizing processes of assembly, persistence, and diversification across the vast Beringian region. BCP collections also represent baseline records of biotic diversity from across the northern high latitudes at a time of accelerated environmental change. These specimens and associated data form an unmatched resource for identifying hidden diversity, interpreting past responses to climate oscillations, documenting contemporary conditions, and anticipating outcomes for complex biological systems in a regime of ecological perturbation. Because of its dual focus on hosts and parasites, the BCP record also provides a foundation for comparative analyses that can document the effects of dynamic change on the geographic distribution, transmission dynamics, and emergence of pathogens. By using specific examples from carnivores, eulipotyphlans, lagomorphs, rodents, ungulates, and their associated parasites, we demonstrate how broad, integrated field collections provide permanent infrastructure that informs policy decisions regarding human impact and the effect of climate change on natural populations.

Neorickettsia (Rickettsiales, Anaplasmataceae) is a genus of obligate intracellular bacterial endosymbionts of digeneans (Platyhelminthes, Digenea). Some Neorickettsia are able to invade cells of the digenean's vertebrate host and are known to cause diseases of domestic animals, wildlife, and humans. In this study we report the results of screening digenean samples for Neorickettsia collected from bats in Egypt and Mindoro Island, Philippines, snails and fishes from Thailand, and fishes from Vietnam and the USA. Neorickettsiae were detected using a real-time PCR protocol targeting a 152 bp fragment of the heat shock protein coding gene, GroEL, and verified with nested PCR and sequencing of a 1853 bp long region of the GroESL operon and a 1371 bp long region of 16S rRNA. Eight unique genotypes of Neorickettsia were obtained from digenean samples. Neorickettsia sp. 8 obtained from Lecithodendrium sp. from Egypt; Neorickettsia sp. 9 and 10 obtained from two species of Paralecithodendrium from Mindoro, Philippines; Neorickettsia sp. 11 from Lecithodendrium sp. and Neorickettsia sp. 4 (previously identified from Saccocoelioides lizae, from China) from Thailand; Neorickettsia sp. 12 from Dicrogaster sp. Florida, USA; Neorickettsia sp. 13 and SF agent from Vietnam. Sequence comparison and phylogenetic analysis demonstrated that the forms, provisionally named Neorickettsia sp. 8-13, represent new genotypes. We have for the first time detected Neorickettsia in a digenean from Egypt (and the African continent as a whole), the Philippines, Thailand and Vietnam based on PCR and sequencing evidence. Our findings suggest that further surveys from the African continent, SE Asia, and the Island countries are likely to reveal new Neorickettsia lineages as well as new digenean host associations.

Members of the genus

Despite being a significant element of biodiversity and playing an important role in their hosts’ physiology, ecology and evolution, parasites remain understudied and are often neglected during biotic surveys and other collection efforts. More often than not, it is due to the perception that parasitological examination requires specialized expertise and substantial additional time to perform. Herein, we provide an express method for recovery and fixation of endoparasites from the gastrointestinal tract of small mammals that can be used by both parasitologists and vertebrate biologists in either field or laboratory conditions. The method does not require special training beyond minimal practice and does not need optics or other special equipment. Our goal is to encourage collection of endoparasites as part of biotic surveys by vertebrate biologists to provide valuable additional biodiversity information in cases when a traditional complete parasitological examination of hosts is not feasible. We believe that this technique will also be useful for professional parasitologists.

We report the first Canadian record of the Holarctic least shrew (Sorex minutissimus Zimmermann, 1780) and associated helminth worms, collected along the Dempster Highway in central Yukon in 2014. We identify the specimen based on morphological characters, characterize the habitat, report other mammals and helminth species associated with this specimen, and use mitochondrial DNA sequences to place the specimen within a phylogenetic context and address Pleistocene refugial hypotheses. Although long considered an Eurasian endemic, the diminutive least shrew was first reported from Alaska in 1994. Our new record for Canada indicates that the species may occur at least as far east as the MacKenzie River and DNA variation suggests this species persisted only in the Beringian refugium in North America during the Last Glacial Maximum. The discovery of a new mammal and associated parasites for Canada points to the urgent need for more detailed information on high-latitude biotas in North America, data that are best obtained through museum-based field surveys, particularly for small, cryptic species.

Bacteria of the genus Neorickettsia are obligate intracellular endosymbionts of parasitic flukes (Digenea) and are passed through the entire complex life cycle of the parasite by vertical transmission. Several species of Neorickettsia are known to cause diseases in domestic animals, wildlife, and humans. Quantitative data on the transmission of the bacteria through the digenean life cycle is almost completely lacking. This study quantified for the first time the abundance of Neorickettsia within multiple stages of the life cycle of the digenean Plagiorchis elegans. Snails Lymnaea stagnalis collected from a pond in North Dakota were screened for the presence of digenean cercariae, which were subsequently tested for the presence of Neorickettsia. Three L. stagnalis were found shedding P. elegans cercariae infected with Neorickettsia. These snails were used to initiate three separate laboratory life cycles and obtain all life cycle stages for bacterial quantification. A quantitative real-time PCR assay targeting the GroEL gene was developed to enumerate Neorickettsia sp. within different stages of the digenean life cycle. The number of bacteria significantly increased throughout all stages, from eggs to adults. The two largest increases in number of bacteria occurred during the period from eggs to cercariae and from 6-day metacercariae to 48-h juvenile worms. These two periods seem to be the most important for Neorickettsia propagation through the complex digenean life cycle and maturation in the definitive host.

The Dicrocoeliidae is a highly diverse family of digeneans parasitic in amniotic tetrapods. Detailed molecular phylogenetic analysis of dicrocoeliids is lacking and only a few dicrocoeliids from mammals have been included in previous studies. Sequence data were previously absent for the Anenterotrematidae that shares several morphological characteristics with dicrocoeliids. We examined phylogenetic affinities of several newly sequenced (nuclear 28S rDNA) taxa of dicrocoeliids and anenterotrematids collected from small mammals in Ecuador, Panama, Peru, USA and Vietnam. Our analyses demonstrated that the two anenterotrematid genera (Anenterotrema, Apharyngotrema) belong to the Dicrocoeliidae, placing the Anenterotrematidae into synonymy with the Dicrocoeliidae. Molecular data combined with morphological examination of type and new specimens provided evidence that Parametadelphis and Apharyngotrema are junior synonyms of Metadelphis, with all Metadelphis species lacking a digestive system. Phylogenetic analysis demonstrates that reduction of the alimentary tract in Lutztrema and its loss in Anenterotrema and Metadelphis represent at least two independent evolutionary events. Genera Brachylecithum, Brachydistomum, and Lyperosomum proved to be non-monophyletic, each likely representing more than a single genus. Furthermore, phylogenetic analysis did not support monophyly of the two largest subfamilies of the Dicrocoeliidae (Dicrocoeliinae and Leipertrematinae) with the other two subfamilies not included in this study. Therefore, we propose to abandon the current subfamily division of the Dicrocoeliidae. Analysis of host associations indicates multiple host-switching events throughout evolution of dicrocoeliids. Lastly, analysis of dicrocoeliid geographic distribution revealed that nearly all major clades included taxa from more than a single zoogeographic realm with the exception of the clade Anenterotrema + Metadelphis, found only in the Neotropics.

The Digenea (Platyhelminthes: Trematoda) are a diverse and complex group of internal metazoan parasites. These parasites can serve as hosts to obligate intracellular bacteria belonging to the genus Neorickettsia (Family: Anaplasmataceae). Neorickettsiae persist within all stages of the fluke life cycle and thus are maintained through vertical transmission. However, the low prevalence of Neorickettsia in nature limits study of their transmission biology at different steps of digenean life cycles. To resolve this dilemma, we have developed for the first time a laboratory model allowing to maintain Neorickettsia sp. through the whole life cycle of a digenean, Plagiorchis elegans. The laboratory life cycle of P. elegans consists of a snail first intermediate host, Lymnaea stagnalis, an aquatic arthropod second intermediate host, Culex pipiens (mosquito larva), and a vertebrate definitive host, Mesocricetus auratus (Syrian hamster). This paper focuses on the development of the laboratory life cycle, as well as outlines its potential uses in studying the transmission biology of Neorickettsia and its evolutionary relationship within its digenean host.

Author(s): Cook, JA; Greiman, SE; Agosta, SJ; Anderson, RP; Arbogast, BS; Baker, RJ; Boeger, W; Bradley, RD; Brooks, DR; Cole, R; Demboski, JR; Dobson, AP; Dunnum, JL; Eckerlin, RP; Esselstyn, J; Galbreath, KE; Hawdon, J; Hoekstra, HE; Kutz, SJ; Light, JE; Olson, LE; Patterson, BD; Patton, JL; Phillips, AJ; Rickart, E; Rogers, DS; Siddall, ME; Tkach, VV; Hoberg, EP

Two new species of Staphylocystoides are described from masked shrews Sorex cinereus. Staphylocystoides oligospinosus n. sp. was collected from the vicinity of Missoula, Montana, United States, and Staphylocystoides parasphenomorphus n. sp. was collected from the vicinity of Whitehorse, Yukon Territory, Canada. Morphological differentiation from known species is provided. Both species are morphologically closest to Staphylocystoides sphenomorphus and to each other. Among other characters, S. oligospinosus n. sp. can be easily differentiated from all known species of the genus by unique cirrus armature, which consists of a short zone of small spines at the base of the cirrus, a few large sparsely distributed spines of varying size in the middle part of the cirrus, and hair-like microtriches densely covering the apical portion of the cirrus. Staphylocystoides parasphenomorphus n. sp. differs from S. oligospinosus n. sp. in a number of characters, most distinctly in cirrus armature, and from another morphologically similar species, S. sphenomorphus, in the number of proglottids, strobila size, number and size of rostellar hooks, and relative length of cirrus sac. Comparison of partial sequences of nuclear large ribosomal RNA subunit gene (1,310 base pairs [bp]) and mitochondrial NAD(P)H dehydrogenase subunit 1 gene (710 bp) strongly supports the status of the described forms as new species. This increases the number of Staphylocystoides species known in North America to 9 (5 parasitizing S. cinereus) and the total number of species in the genus to 12. Staphylocystoides parasphenomorphus n. sp. is the first named Staphylocystoides record in Canada.

Ixodes angustus (Neumann) (Acari: Ixodidae) is considered to be a nidicolous tick in that the entire life cycle can be completed in the host nest. Males of this tick have been reported to be rare on hosts because most mating occurs in the host nest and males typically do not feed on hosts. Collections of I. angustus in Alaska departed slightly from this paradigm in that nine males were collected from hosts, mostly in copulation with females that were attached to mammalian hosts. Non-nidicolous mating was therefore more common in I. angustus collected in this study than has been reported previously. A hyperparasitic male I. angustus was found firmly attached via its hypostome and chelicerae to the ventral idiosoma of a partially engorged female I. angustus that was attached to, and feeding on, an American red squirrel, Tamiasciurus hudsonicus (Erxleben). This hyperparasitic interaction is discussed and illustrated with a Scanning Electron Micrograph.

Staphylocystoides gulyaevi n. sp. is described based on specimens obtained from the dusky shrew Sorex monticolus collected on Sukkwan Island, southeast Alaska. Staphylocystoides gulyaevi n. sp. is compared with other North American members of the genus having 10 rostellar hooks. The new species is morphologically similar to Staphylocystoides parvissima and Staphylocystoides asketus. The uterus in S. gulyaevi n. sp. develops much more rapidly, and a well-developed uterus appears abruptly after it is barely visible in a previous proglottid. In S. parvissima the uterus grows gradually, and its early development is seen in several proglottids. At the level of pre-gravid proglottids the uterus of S. gulyaevi n. sp. occupies only the middle field of the proglottid, while in S. parvissima it fills the whole proglottid including lateral fields. The rostellar hooks in the new species are significantly smaller in size than in S. asketus. Additionally, the new species has fewer proglottids than S. asketus, while having a similar strobila length. Molecular comparison, using 3 genes (28s rDNA, cox1, and nad1), between S. gulyaevi n. sp. and S. parvissima, further corroborates the status of S. gulyaevi n. sp. as a new species. The new species is the seventh species of Staphylocystoides found in North America and the first cestode or any helminth reported from shrews in southeast Alaska.

The ultrastructure of the mature spermatozoon of the type genus of the Plagiorchiidae Plagiorchis elegans (Rudolphi, 1802), a parasite of the Golden hamster, Mesocricetus auratus is described. This study is the first ultrastructural study of the spermatozoon of a Plagiorchis, the second of a plagiorchiid species and only the third in the Plagiorchioidea. Previously data on spermatozoon ultrastructure existed only for the plagiorchiid Enodiotrema reductum and the omphalometrid Rubenstrema exasperatum. The mature spermatozoon of P. elegans exhibited the general pattern described in most digenean species, namely two axonemes of the 9 + "1" Trepaxonemata pattern, nucleus, mitochondria, external ornamentation of the plasma membrane, spine-like bodies, and glycogen granules. However, the rather typical expansion of the plasma membrane is not found in P. elegans. Another peculiarity of the spermatozoon of P. elegans is the presence of a structure called thin cytoplasm termination. Spermatozoon ultrastructure of P. elegans is compared with that of E. reductum and R. exasperatum. Spermatozoon of P. elegans conforms to the general pattern described in E. reductum. Thus, this study further expands our knowledge on the spermatozoon ultrastructure among the members of the Plagiorchioidea, one of the most phylogenetically derived groups of the digenea.

The trematode Nanophyetus salmincola is known as the carrier of Neorickettsia helminthoeca, an obligate intracellular endosymbiotic bacterium that causes salmon poisoning disease (SPD), a fatal disease of dogs. The bacteria are maintained through the complex life cycle of N. salmincola that involves snails Juga plicifera as the first intermediate host, salmonid fishes as the second intermediate host and fish-eating mammals as definitive hosts. N. salmincola was also found to harbor a second species of Neorickettsia that causes the Elokomin fluke fever disease (EFF) which has clinical signs similar to SPD in bears, but only low grade illness in dogs. The EFF agent has not been sequenced. In this study we identified N. salmincola as the vector of yet additional species of Neorickettsia known as Stellanchasmus falcatu (SF) agent using DNA sequencing.