Resolution of Clinical Signs of Sarcoptic Mange in American Black Bears (Ursus americanus), in Ivermectin-Treated and Nontreated Individuals.

The parasitic mite Sarcoptes scabiei causes mange in nearly 150 species of mammals by burrowing under the skin, triggering hypersensitivity responses that can alter animals' behavior and result in extreme weight loss, secondary infections, and even death. Since the 1990s, sarcoptic mange has increased in incidence and geographic distribution in Pennsylvania black bear (Ursus americanus) populations, including expansion into other states. Recovery from mange in free-ranging wildlife has rarely been evaluated. Following the Pennsylvania Game Commission's standard operating procedures at the time of the study, treatment consisted of one subcutaneous injection of ivermectin. To evaluate black bear survival and recovery from mange, from 2018 to 2020 we fitted 61 bears, including 43 with mange, with GPS collars to track their movements and recovery. Bears were collared in triplicates according to sex and habitat, consisting of one bear without mange (healthy control), one scabietic bear treated with ivermectin when collared, and one untreated scabietic bear. Bears were reevaluated for signs of mange during annual den visits, if recaptured during the study period, and after mortality events. Disease status and recovery from mange was determined based on outward gross appearance and presence of S. scabiei mites from skin scrapes. Of the 36 scabietic bears with known recovery status, 81% fully recovered regardless of treatment, with 88% recovered with treatment and 74% recovered without treatment. All bears with no, low, or moderate mite burdens (<16 mites on skin scrapes) fully recovered from mange (n=20), and nearly half of bears with severe mite burden (≥16 mites) fully recovered (n=5, 42%). However, nonrecovered status did not indicate mortality, and mange-related mortality was infrequent. Most bears were able to recover from mange irrespective of treatment, potentially indicating a need for reevaluation of the mange wildlife management paradigm.

Evaluation of the immunoprotective effects of eight recombinant proteins from Baylisascaris schroederi in mice model.

BACKGROUND: Baylisascaris schroederi is the most common and harmful intestinal parasitic nematode of giant pandas, causing ascariasis. Although drug deworming is the main measure to control ascariasis in captive giant pandas, prolonged and repeated use of deworming drugs might induce resistance in nematodes and drug residues in giant pandas. Therefore, developing a safe and effective vaccine might provide a novel strategy to prevent ascariasis in captive giant pandas. METHODS: Four highly expressed secretome genes encoding excretory and secretory proteins of B. schroederi, including transthyretin-like protein 46 (BsTLP), uncharacterized protein (BsUP), hypothetical protein 1 (BsHP1), and hypothetical protein 2 (BsHP2) and four functional genes [(encoding Galectin (BsGAL), glutathione S-transferase (BsGST), fatty acid-binding protein (BsFABP), and thioredoxin peroxidase (BsTPX)] were identified based on genome and transcriptome databases of B. schroederi and used to construct recombinant proteins via prokaryotic expression. Kunming mice were vaccinated subcutaneously twice with the recombinant proteins (50 µg/mouse) mixed with Quil A adjuvant with a 2-week interval and then orally challenged with 3000 infective eggs. The immunoprotective effects of the eight recombinant proteins on mice were assessed comprehensively using surface lesion histology scores of the mouse liver and lung, larval worm reduction, serum antibody levels (IgG, IgE, IgA, IgG1, and IgG2a), and cytokine production [interferon gamma (IFN-γ), interleukin (IL)-2, IL-4, IL-5, and IL-10]. RESULTS: Mice vaccinated with recombinant (r)BsUP (76.5%), rBsGAL (74.7%), and rBsHP2 (71.5%) showed a significant (P < 0.001) reduction in the larval worm rate compared with that in the adjuvant control. Besides, the surface lesions in the liver and lung of the vaccinated mice were alleviated. Serum levels of total IgG, IgE, IgA, IgG1, IgG2a, and cytokines, including IL-10, IL-5, and IFN-γ, were significantly higher (P < 0.001) than those in the control group. CONCLUSIONS: The results showed that candidate three vaccines (rBsUP, rBsGAL, and rBsHP2) could provide effective protection against egg infection in mice associated with a mixed Th1/2-type immune response.

USE OF A POINT OF CARE TEST TO DETERMINE THE PREVALENCE OF ANTIBODIES TO TOXOPLASMA GONDII IN BLACK BEARS FROM NORTH CAROLINA AND PENNSYLVANIA.

Toxoplasma gondii is an important protozoan parasite of humans and animals throughout the world. Black bears are among the animals with the highest seroprevalence of T. gondii in the United States. A rapid point of care (POC) test is commercially available to detect antibodies to T. gondii in humans. We evaluated the utility of the POC test to detect anti-T. gondii antibodies in 100 wild black bears from North Carolina (n = 50) and Pennsylvania (n = 50). In a blind study, sera were tested by the POC test, and results were compared to the modified agglutination test (MAT). Overall, anti-T. gondii antibodies were detected in 76% (76/100) black bears by both MAT and POC tests. One false positive and one false negative result in the POC test were obtained in bears from Pennsylvania. The sensitivity and specificity of the POC test were both 99% when compared to the MAT. Results from our study indicate the POC test could be a useful screening tool for serological surveillance of T. gondii in black bears.

Further studies on the morphology of Baylisascaris transfuga (Rudolphi, 1819) (Nematoda: Ascaridomorpha: Ascarididae) from the polar bear Ursus maritimus Phipps (Carnivora: Ursidae).

Baylisascaris transfuga (Rudolphi, 1819) is a common parasitic nematode in the digestive tract of various species of bears worldwide, with great veterinary significance. However, our present knowledge on the morphology of B. transfuga remains insufficient. In the present study, the detailed morphology of B. transfuga was studied using light and scanning electron microscopy (SEM), based on specimens collected from the polar bear Ursus maritimus Phipps (Carnivora: Ursidae) in the Shijiazhuang Zoo, China. The results revealed some morphological and morphometric variation between the present specimens and some of those from previous studies, including oesophageal length of female, number and morphology of postcloacal papillae and morphology of tail of males. Present SEM observations clearly showed the detailed morphology of lips, cervical alae, cloacal ornamentation, precloacal medioventral papilla, phasmids and tail tip. These supplementary morphological and morphometric data enable us to identify this ascaridid nematode more accurately.

Survey for Selected Parasites in Alaska Brown Bears (Ursus arctos).

To assess infection with or exposure to endo- and ectoparasites in Alaska brown bears (Ursus arctos), blood and fecal samples were collected during 2013-17 from five locations: Gates of the Arctic National Park and Preserve; Katmai National Park; Lake Clark National Park and Preserve; Yakutat Forelands; and Kodiak Island. Standard fecal centrifugal flotation was used to screen for gastrointestinal parasites, molecular techniques were used to test blood for the presence of Bartonella and Babesia spp., and an ELISA was used to detect antibodies reactive to Sarcoptes scabiei, a species of mite recently associated with mange in American black bears (Ursus americanus). From fecal flotations (n=160), we identified the following helminth eggs: Uncinaria sp. (n=16, 10.0%), Baylisascaris sp. (n=5, 3.1%), Dibothriocephalus sp. (n=2, 1.2%), and taeniid-type eggs (n=1, 0.6%). Molecular screening for intraerythrocytic parasites (Babesia spp.) and intracellular bacteria (Bartonella spp.) was negative for all bears tested. We detected antibodies to S. scabiei in six of 59 (10.2%) individuals. The relatively low level of parasite detection in this study meets expectations for brown bear populations living in large, relatively undisturbed habitats near the northern edge of the range. These results provide a contemporary understanding of parasites in Alaska brown bears and establish baseline levels of parasite presence to monitor for changes over time and relative to ecologic alterations.

Morphological and molecular phylogenetical identification of Tricodectes pinguis from Japanese black bears (Ursus thibetanus japonicus) in Aomori Prefecture, Japan.

Trichodectes pinguis, referred to commonly as the bear-biting louse, has been reported in several bear species. However, graphical (blurred or coarse) and genetic information on the louse is limited. In this study, we identified T. pinguis collected from Japanese black bears in the Aomori Prefecture, Japan. We confirmed 12S rDNA sequences derived from the collected T. pinguis and performed molecular phylogenetic analysis based on 12S rDNA. The analysis revealed the parasitic louse to be T. pinguis. Interestingly, the body size of T. pinguis found in this study was smaller than the previous recorded body size of them in Japan and Turkey. To better understand the biting louse infesting bears, morphometric and genetic information from other bear hosts needs to be accumulated.

MORBIDITY AND MORTALITY OF FREE-RANGING AMERICAN BLACK BEARS (URSUS AMERICANUS) UNDERGOING REHABILITATION IN EASTERN TENNESSEE, USA, 1996-2021.

Free-ranging American black bears (Ursus americanus) often share habitat with humans and domestic animals, predisposing them to anthropogenic conflicts. Rehabilitation under professional care is a management option for orphaned, injured, and/or ill bears. Across several southeastern states, rescued bears are assessed and treated at the University of Tennessee and rehabilitated at Appalachian Bear Rescue (ABR). Records from 1996-2021 showed 337 bears (170 males, 166 females, 1 unknown) from nine states were admitted to ABR. Three bears were admitted twice, resulting in 340 admissions (42 neonates <3 mo old, 206 cubs 3-12 mo, 87 yearlings 1-2 yr, and 5 adults >2 yr). Bears presented as orphans (58%), malnourished (24%), injured or ill (12%), or confiscated/other (6%). Individuals were returned to the wild (85%); died or were euthanized (12%); or were placed into professional care (3%). Of released bears, 195 had complete medical records available for evaluation; 31% were healthy upon intake while the remaining were treated successfully for malnutrition and internal parasites (49%), orthopedic (9%) and soft tissue injuries (5%), or other diseases (5%). Causes of death determined during necropsies performed (n=30) were classified as trauma (50%), developmental (13%), undetermined (13%), malnutrition (13%), infectious or inflammatory (7%), and toxicosis (3%). Despite the lack of maternal care and high prevalence of malnutrition and trauma, most bears recovered to release with appropriate husbandry and medical care. This study provides a foundation for research to further improve care of rehabilitating black bears.

Preliminary findings on the gastrointestinal parasites of the brown bear (Ursus arctos) in the Cantabrian mountains, Spain.

No study is currently available on the parasitofauna of the population of brown bears (Ursus arctos) inhabiting the Cantabrian Mountains in Spain. The aim of the present study was to obtain novel information on diversity and prevalence of gastrointestinal parasites in these individuals. During August 2016 and from May to July 2017, 14 fecal samples were collected from the western Cantabrian bear subpopulation, in the Somiedo Natural Park, in the Spanish province of Asturias. The prevalence of parasites detected was 71% and two genera were identified: Dicrocoelium sp. and Trichuris sp. Since the impact that pathogens such as endoparasites can have on the health of bears, together with other stressors, is still poorly understood, research efforts that include disease surveillance are critical to the successful protection of this emblematic species. Our preliminary findings require further investigations, with a wider sampling effort, and bring awareness for the need of carrying further studies on this area as a part of a proactive species management plan.

Pelodera strongyloides in the critically endangered Apennine brown bear (Ursus arctos marsicanus).

Skin biopsies from 20 Apennine brown bears (Ursus arctos marsicanus), 17 of which displaying skin lesions, were investigated by histopathology. Different degrees of dermatitis characterized by folliculitis and furunculosis accompanied by epidermal hyperplasia and epidermal and follicular hyperkeratosis were detected. In the most severe lesions, the superimposition of traumatic wounds, probably self-induced by scratching, was observed. In 8 out of 17 (47.0%) affected bears, cross- and longitudinally-sectioned nematode larvae were present within the lumen of hair follicles, whose localization and morphological characteristics were consistent with Pelodera strongyloides. P. strongyloides is a free-living saprophytic nematode whose third-stage larvae can invade the skin causing pruritic dermatitis in several mammalian species. This is the first report of Pelodera infection in the brown bear. Although capable of causing primary dermatitis, the finding of Pelodera is not sufficient to conclude that it is the cause of the lesions observed in bears. Nevertheless, the high prevalence of the infection is indicative of a diffuse phenomenon that requires further specific investigations given the interest and conservational relevance of this relict bear population.

Genome of the Giant Panda Roundworm Illuminates Its Host Shift and Parasitic Adaptation.

Baylisascaris schroederi, a roundworm (ascaridoid) parasite specific to the bamboo-feeding giant panda (Ailuropoda melanoleuca), represents a leading cause of mortality in wild giant panda populations. Here, we present a 293-megabase chromosome-level genome assembly of B. schroederi to infer its biology, including host adaptations. Comparative genomics revealed an evolutionary trajectory accompanied by host-shift events in ascaridoid parasite lineages after host separations, suggesting their potential for transmission and rapid adaptation to new hosts. Genomic and anatomical lines of evidence, including expansion and positive selection of genes related to the cuticle and basal metabolisms, indicate that B. schroederi undergoes specific adaptations to survive in the sharp-edged bamboo-enriched gut of giant pandas by structurally increasing its cuticle thickness and efficiently utilizing host nutrients through gut parasitism. Additionally, we characterized the secretome of B. schroederi and predicted potential drug and vaccine targets for new control strategies. Overall, this genome resource provides new insights into the host adaptation of B. schroederi to the giant panda as well as the host-shift events in ascaridoid parasite lineages. Our findings on the unique biology of B. schroederi will also aid in the development of prevention and treatment measures to protect giant panda populations from roundworm parasitism.

INVESTIGATION OF SARCOCYSTIS SPP. INFECTION IN FREE-RANGING AMERICAN BLACK BEARS (URSUS AMERICANUS) AND GRIZZLY BEARS (URSUS ARCTOS HORRIBILIS) IN BRITISH COLUMBIA, CANADA.

Sarcocystis spp. are protozoan parasites that cause a spectrum of lesions in various hosts. Hepatic sarcocystosis and encephalitis have been described in captive American black bears (Ursus americanus) and polar bears (Ursus maritimus), and in a free-ranging grizzly bear (Ursus arctos horribilis), but have not previously been reported in free-ranging American black bears. This study aimed to characterize the presence and lesions associated with Sarcocystis spp. in free-ranging bears in British Columbia, Canada from samples submitted to the provincial diagnostic laboratory. From 2007 to 2019, 102 free-ranging American black bear and grizzly bear tissues were examined postmortem for sarcocystosis using histopathology and follow-up molecular diagnostics. Sarcocystosis was confirmed in 41 (40%) free-ranging bears including 39 American black bears and two grizzly bears. Microscopic lesions included multifocal necrotizing hepatitis, nonsuppurative encephalitis, and/or intramuscular sarcocysts with or without associated inflammation. Sarcocystosis was considered the cause of death in eight (20%) of these bears, exclusively in cubs of the year (<1 yr old). Sarcocystis canis was identified in 22/32 (69%) cases where molecular characterization was performed and was the etiologic agent associated with bears that died of sarcocystosis. Confirmed cases were distributed widely across British Columbia. While there was an alternate proximate cause of death in the other confirmed bears, sarcocystosis may have contributed. Age was a significant risk factor, with yearlings presenting more often with fulminant lesions; however, there was a sampling bias toward juvenile bear submissions due to size and ease of transport. Further research is needed to understand the disease epidemiology and significance to population health.

EPIDEMIOLOGIC AND PUBLIC HEALTH SIGNIFICANCE OF TOXOPLASMA GONDII INFECTIONS IN BEARS (URSUS SPP.): A 50 YEAR REVIEW INCLUDING RECENT GENETIC EVIDENCE.

Toxoplasma gondii infections are common in humans and animals worldwide. The present review summarizes worldwide information on the prevalence of clinical and subclinical infections, epidemiology, and genetic diversity of T. gondii infections in bears. Seroprevalence estimates of T. gondii in black bears (Ursus americanus) are one of the highest of all animals. In Pennsylvania, seroprevalence is around 80% and has remained stable for the past 4 decades. Approximately 3,500 bears are hunted yearly in Pennsylvania alone. The validity of different serological tests is discussed based on bioassay and serological comparisons. Seroprevalence in grizzly bears (Ursus arctos) is lower than that in black bears. Even polar bears (Ursus maritimus) are infected; infections in these animals are ecologically interesting because of the absence of felids in the Arctic. Clinical toxoplasmosis in bears is rare and not documented in adult animals. The few reports of fatal toxoplasmosis in young bears need confirmation. Viable T. gondii has been isolated from black bears and a grizzly bear. The genetic diversity of isolates based on DNA from viable T. gondii isolates is discussed. Genetic typing of a total of 26 T. gondii samples from bears using 10 PCR-RFLP markers revealed 8 PCR-RFLP ToxoDB genotypes: #1 (clonal type II) in 3 samples, #2 (clonal type III) in 8 samples, #4 (haplogroup 12) in 3 samples, #5 (haplogroup 12) in 3 samples, #74 in 5 samples, #90 in 1 sample, #147 in 1 sample, and #216 in 2 samples. These results suggest relatively high genetic diversity of T. gondii in bears. Overall, T. gondii isolates in bears range from those circulating in a domestic cycle (genotypes #1 and #2) to those mainly associated with wildlife (such as genotypes #4 and #5, together known as haplogroup 12). A patient who acquired clinical Trichinella spiralis infection after eating undercooked bear meat also acquired T. gondii infection. Freezing of infected meat kills T. gondii, including the strains isolated from bears.

Seroprevalence of Toxoplasma gondii in American Black Bears (Ursus americanus) in Nevada, USA, using an Enzyme-linked Immunosorbent Assay.

Archived serum samples taken between 1997 and 2017 from 170 American black bears (Ursus americanus) in the Lake Tahoe area between California and Nevada, US, were tested for Toxoplasma antibodies to assess the seroprevalence of this agent. Samples were screened using a commercial porcine Toxoplasma (enzyme-linked immunosorbent assay [ELISA]) modified with Protein A/G peroxidase and compared to a traditional fluorescent antibody test. Results were analyzed to determine if there were differences in seroprevalence based on the test used, sex of bears, or habitat usage (urban-suburban vs. wildland). No significant differences in seroprevalence were attributable to any of these parameters. The overall seropositivity for bears was 36% (62/170), with urban-suburban bears scoring lower (31%; 37/119) than rural-wildland bears (40%; 18/45). Our results strongly support the use of a Protein A/G-modified ELISA for determining Toxoplasma exposure in black bears. We found somewhat lower levels of Toxoplasma antibodies in black bears from this region than in several reports from populations in the eastern US.

ENDOPARASITES OF FORMOSAN BLACK BEARS (URSUS THIBETANUS FORMOSANUS) DURING ACORN SEASON IN YUSHAN NATIONAL PARK, TAIWAN.

Parasite infection is one of the most important factors in wildlife conservation. However, fecal parasite profiles of threatened Asiatic black bears (Ursus thibetanus) are only sporadically reported, and the effect of parasitic diseases on the survival of the locally endangered Formosan black bear (Ursus thibetanus formosanus) in Taiwan remains undetermined. The study objective was to investigate the gastrointestinal parasite profiles of Formosan black bears in Yushan National Park, the only known high-density habitat for the species in Taiwan. Bear fecal samples were collected in the acorn season (from October to February) from January 2008 to October 2012. To avoid bias created by repeat sampling, the parasite profiles of fecal samples collected in 2010 from 46 individually identified bears (which were identified by genetic analysis) were also examined. Parasites were isolated by various methods and identified by morphologic characteristics. A total of 220 samples were analyzed and the results were compared between seasons, sexes, and individuals. The overall frequency of parasite infection was 77.3%, and it varied by species, with Baylisascaris transfuga infection being the most frequent. We suggest that one factor underlying the high frequency and high intensity of infection that we observed is the fact that the bears seasonally congregated at high density in a small area. To our knowledge, this is the first thorough study of gastrointestinal parasites in Asiatic black bears. The long-term nature of the investigation and the relatively stable frequency and intensity of infection suggest that parasitic diseases could serve as bioindicators of ecosystem health.

High Prevalence and Intensity of Trichinella Infection in Yukon American Black (Ursus americanus) and Grizzly (Ursus arctos) Bears.

Trichinella spp. nematodes are commonly found in bear species (Ursidae) and can pose severe health risks to humans when infective first-stage larvae are ingested in meat. Samples of tongue or masseter muscle from 22 male and 11 female American black bears (Ursus americanus; mean age 6.5 yr, range 1-16 yr) and 22 male, eight female, and one unknown sex grizzly bears (Ursus arctos; mean age 8.8 yr, range 2-28 yr), from Yukon, Canada, were tested to determine prevalence and intensity of Trichinella spp. infection. For black bears, prevalence was 20% and mean intensity was 401 larvae per gram of tissue (LPG), whereas for grizzly bears, prevalence was 71%, and mean infection intensity was 35 LPG. Isolates from all positive samples were identified as genotype Trichinella-T6 by multiplex PCR. For black bears, prevalence is the highest reported in Canada and infection intensity the highest recorded in North America. One black bear had a larval burden of 1,173 LPG, the second highest recorded in any host species. The prevalence in grizzly bears was the highest reported in Canada for this host. In total, 90% (27 of 30) of infected bears had infection burdens above the human food safety threshold of ≥1 LPG, reinforcing the importance of communicating the health risks to people consuming bear meat.

Molecular detection of Blastocystis in black bears and sika deer in northern China.

Protists of the Blastocystis genus are distributed worldwide and can infect a range of hosts. However, data concerning Blastocystis infection are limited for sika deer and are not available for black bears. Therefore, in the present study, a total of 312 black bears (Ursus thibetanus) from Heilongjiang Province and 760 sika deer (Cervus nippon) from four different northern Chinese provinces were investigated. Blastocystis infection in these animals was detected via PCR amplification of the small subunit rRNA gene in fecal samples. The prevalence of Blastocystis infection in black bears and sika deer was 14.4% (45/312 positive samples) and 0.8% (6/760 positive samples), respectively. Young black bears (18.3%) had a significantly higher Blastocystis prevalence than adult bears (9.1%). The prevalence of Blastocystis was significantly higher in black bears raised outdoors (24.6%) than in bears raised indoors (12.2%). Blastocystis-positive sika deer were only found in Jilin Province (1.3%, 6/480). Female sika deer (0%, 0/61) had a significantly lower Blastocystis prevalence than males (0.9%, 6/699). Sanger sequencing was used to determine the small subunit rRNA gene sequences of the Blastocystis-positive PCR products. A neighbor-joining phylogenetic tree based on the small subunit rRNA gene sequences showed that only Blastocystis subtype (ST)1 was identified in black bears, whereas ST10 and ST14 were found in sika deer. This is the first report of Blastocystis ST1 infection in black bears. These findings also extend the distribution information of Blastocystis subtypes, which will provide a foundation for further study of Blastocystis in different hosts in China.

Occurrence and genetic diversity of Cryptosporidium spp. in wild foxes, wolves, jackals, and bears in central Europe.

Parasites of the genus Cryptosporidium Tyzzer, 1910 are one of the most common protistan parasites of vertebrates. Faecal samples from 179 red foxes (Vulpes vulpes [Linnaeus]), 100 grey wolves (Canis lupus Linnaeus), 11 golden jackals (Canis aureus Linnaeus), and 63 brown bears (Ursus arctos Linnaeus) were collected in the Czech Republic, Poland and Slovakia. Samples were examined for the presence of Cryptosporidium spp. using microscopy and PCR/sequence analysis. Phylogenetic analysis based on the small subunit ribosomal RNA (SSU), actin and 60-kDa glycoprotein (gp60) genes using the maximum likelihood method revealed the presence of Cryptosporidium tyzzeri Ren, Zhao, Zhang, Ning, Jian et al., 2012 (n = 1) and C. andersoni Lindsay, Upton, Owens, Morgan, Mead et Blackburn, 2000 (n = 2) in red foxes, C. canis Fayer, Trout, Xiao, Morgan, Lai et Dubey, 2001 (n = 2) and C. ubiquitum Fayer, Santín et Macarisin, 2010 (n = 2) in grey wolves, and C. galli Pavlásek, 1999 in brown bears (n = 1) and red foxes (n = 1). Subtyping of isolates of C. ubiquitum and C. tyzzeri based on sequence analysis of gp60 showed that they belong to the XIId and IXa families, respectively. The presence of specific DNA of C. tyzzeri, C. andersoni and C. galli, which primarily infect the prey of carnivores, is probably the result of their passage through the gastrointestinal tract of the carnivores. Finding C. ubiquitum XIId in wolves may mean broadening the host spectrum of this subtype, but it remains possible this is the result of infected prey passing through the wolf - in this case deer, which is a common host of this parasite. The dog genotype of C. canis was reported for the first time in wolves.

A first report of Thelazia callipaeda infection in Phortica okadai and wildlife in national nature reserves in China.

BACKGROUND: Thelazia callipaeda is a zoonotic parasitic nematode of the family Thelaziidae, with Phortica okadai as its intermediate host and only confirmed vector in China. China has the largest number of human cases of thelaziosis in the world. It is generally believed that infected domestic animals (dogs and cats) are the most important reservoir hosts of T. callipaeda, and thus pose a direct threat to humans. At present, there is little research or attention focused on the role of wildlife in the transmission cycle of thelaziosis in nature reserves. METHODS: We selected locations in four national nature reserves across China to monitor P. okadai and wildlife. We used a fly-trap method to monitor P. okadai density. Morphological analysis of the parasites collected from the conjunctival sac of the infected wildlife was undertaken as the first step in species identification, and polymerase chain reaction (PCR) was used for species confirmation. RESULTS: In 2019, the density of P. okadai in Foping National Nature Reserve in China increased sharply, and infected P. okadai were newly found in the reserve. Giant panda, wild boar, leopard cat, and black bear were found to be newly infected with T. callipaeda (one individual of each species). A total of four worms were collected, one from each species of wildlife. The four worms were identified as T. callipaeda by their morphological characteristics; species identification was confirmed by PCR amplification. CONCLUSIONS: To the best of our knowledge, this is the first report of T. callipaeda infection in P. okadai as well as in a variety of wildlife, including giant panda, in nature reserves in China. These results indicate that there is a transmission cycle of T. callipaeda among wildlife in these nature reserves. The increasing number of case reports of thelaziosis in wildlife suggest a likely risk of T. callipaeda infection for the inhabitants of villages situated around nature reserves.

Molecular detection of filarial nematode parasites in Japanese black bears (Ursus thibetanus japonicus) from Iwate Prefecture, Japan.

This study aimed to detect filarial parasites in blood samples of Japanese black bears (Ursus thibetanus japonicus) collected from Iwate Prefecture, Japan. Positive amplicons were obtained from 26 out of 30 samples by nested PCR targeting 18S ribosomal RNA gene and first internal transcribed spacer regions. DNA sequences of Mansonella sp. close to M. ozzardi and Dirofilaria sp. were detected for eight and 11 positive amplicons, respectively. Co-infection was detected for the remaining seven amplicons. Dirofilaria sp. was identified as D. ursi by further genetic analysis of 5S ribosomal RNA gene sequence. The results of this study will contribute to further investigations of Japanese black bears for monitoring their risk as a reservoir of possible zoonotic filarial parasites.

Genomic Signatures of Coevolution between Nonmodel Mammals and Parasitic Roundworms.

Antagonistic coevolution between host and parasite drives species evolution. However, most of the studies only focus on parasitism adaptation and do not explore the coevolution mechanisms from the perspective of both host and parasite. Here, through the de novo sequencing and assembly of the genomes of giant panda roundworm, red panda roundworm, and lion roundworm parasitic on tiger, we investigated the genomic mechanisms of coevolution between nonmodel mammals and their parasitic roundworms and those of roundworm parasitism in general. The genome-wide phylogeny revealed that these parasitic roundworms have not phylogenetically coevolved with their hosts. The CTSZ and prolyl 4-hydroxylase subunit beta (P4HB) immunoregulatory proteins played a central role in protein interaction between mammals and parasitic roundworms. The gene tree comparison identified that seven pairs of interactive proteins had consistent phylogenetic topology, suggesting their coevolution during host-parasite interaction. These coevolutionary proteins were particularly relevant to immune response. In addition, we found that the roundworms of both pandas exhibited higher proportions of metallopeptidase genes, and some positively selected genes were highly related to their larvae's fast development. Our findings provide novel insights into the genetic mechanisms of coevolution between nonmodel mammals and parasites and offer the valuable genomic resources for scientific ascariasis prevention in both pandas.