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Co-infections of mammalian hosts with intestinal helminths and bacterial pathogens are common, especially in areas with inadequate sanitation. Interactions between co-infecting species and host microbiota can cause significant changes in host immunity, disease severity, and pathogen transmission, requiring unique treatment for each case. A greater understanding of the influences of parasite-bacteria co-infections will improve diagnosis and therapeutic approaches to control infectious diseases. To study the influence of the trematode parasite Echinostoma caproni on commensal and pathogenic bacteria in the mouse gut, we examined the abundance of intestinal lactic acid bacteria and Salmonella enterica serovar Typhimurium in control mice not exposed to E. caproni (P–) or S. Typhimurium (S–), E. caproni–infected (P+S–), S. Typhimurium-infected (P –S+), and E. caproni–S. Typhimurium co-infected (P+S+) mice, and determined bacterial burdens in the livers and spleens of the P–S+ and P+S+ mice. We also examined a subset of P+S– and P+S+ mice for survival and the relative location of E. caproni in the small intestine. The numbers of presumptive lactic acid bacteria were significantly higher in the P+S+ and P–S+ mice compared to the uninfected mice, and S. Typhimurium colonization in the liver and spleen was significantly reduced in the P+S+ mice compared to the P–S+ mice. Echinostoma caproni were located anteriorly in the intestine of P+S– mice, while in the P+S+ mice, the parasites were distributed more posteriorly. Survival of E. caproni was unaffected in either group. The results of our study suggest that E. caproni facilitates a higher abundance of presumptive lactic acid bacteria in the mouse intestine and reduces colonization of S. Typhimurium in the liver and spleen of the co-infected host.
Two new species of Viannaia from the intestine of the North American opossums, Didelphis virginiana (Virginia opossum), and Philander opossum (gray four-eyed opossum), are described based on morphological and molecular data, through an integrative taxonomic approach. Maximum likelihood and Bayesian inference analyses for each dataset and the concatenated dataset were performed using a mitochondrial cytochrome c oxidase subunit 1 (COI) gene, and the nuclear ribosomal internal transcribed spacer region (ITS1–5.8S–ITS2). The phylogenetic analyses revealed 2 new species that occur in Mexico, one from the western state of Colima and another from the southern state of Chiapas. Our phylogenetic trees for both molecular markers and concatenated datasets yielded similar topologies with high bootstrap values and posterior probabilities. Viannaia is recovered as a monophyletic group, but the family Viannaiidae appears as non-monophyletic, due to the position of Travassostrongylus scheibelorum, similar to previous studies. Finally, the morphology of Viannaia and Hoineffia is discussed.
Jorrell Fredericks, Dolores E. Hill, Diane S. Hawkins-Cooper, Valsin M. Fournet, Jonathan Calero-Landa, Brandon Adams, Alexis Johnson, Marwa Barrow, Jovan Aquino, Talatha Mahmoud, Victoria Murphy, Alec Barlow, Priyal Patel, Megan George, Nadya Chehab, Matthew Kramer, Nate E. Bauer
Foodborne pathogens continue to pose a public health risk and can cause serious illness and outbreaks of disease in consumers. The consumption of raw or undercooked infected meat, such as pork containing infectious stages of Toxoplasma gondii, may be a major route of transmission to humans. Given the occasional presence of T. gondii in pork meat and the frequent use of pork for products not intended to be cooked, such as dry-cured ham, a potential risk exists for T. gondii transmission to consumers of these products. The purpose of this study was to determine the seroprevalence of T. gondii in U.S. market hogs and sows at slaughter. A total of 20,209 sera samples collected from 22 U.S. slaughterhouses, including 15 of the top 25 largest slaughter plants in the United States, were tested for T. gondii antibodies using a commercial ELISA assay. Seroprevalence in this study was 0.74%, with a herd prevalence of 10.86%. We compared seroprevalence of T. gondii in market hogs vs. sows from a separate but geographically similar set of slaughterhouse locations, with serum samples screened using the T. gondii modified agglutination test. This set of market hogs demonstrated 0% seroprevalence for T. gondii, while sows from geographically similar but separate slaughter facilities demonstrated a seroprevalence of 1.03%. Overall, both analyses show low seroprevalence of T. gondii in U.S market hogs and sows, respectively, and a marked drop in prevalence in market hogs and sows compared to previous studies.
Sequences of the mitochondrial cytochrome c oxidase 1 (COI) gene of 115 Baylisascaris procyonis individuals from 13 U.S. states and 1 Canadian province were obtained from 44 raccoon hosts to assess genetic variation and geographic structure. The maximum genetic distance between individuals was low (1.6%), consistent with a single species. Moderate COI haplotype (h = 0.60) and nucleotide (π = 0.0053) diversity were found overall. Low haplotype diversity was found among samples east of the Mississippi River (h = 0.036), suggesting that historical growth and expansion of raccoon populations in this region could be responsible for high parasite gene flow or a selective sweep of B. procyonis mtDNA. There was low genetic structure (average Φst = 0.07) for samples east of the continental divide, but samples from Colorado showed higher diversity and differentiation from midwestern and eastern samples. There was marked genetic structure between samples from east and west of the continental divide, with no haplotypes shared between these regions. There was no significant isolation by distance among any of these geographic samples. The phylogeographic patterns for B. procyonis are similar to genetic results reported for their raccoon definitive hosts. The phylogeographic divergence of B. procyonis from east and west of the continental divide may involve vicariance resulting from Pleistocene glaciation and associated climate variation.
Snails identified as Triodopsis hopetonensis were collected (n = 18) from the University of Arkansas–Fayetteville campus in December 2018. Additional snails were collected in April 2019 (n = 9) and in September 2019 (n = 9). Kidney tissues were examined using light microscopy, and polysporocystic oocysts were observed. Sporulated oocysts (n = 2) measured 78 ± 3.4 µm × 76 ± 2.9 µm with an irregular oocyst residuum and contained an estimated 44–55 tetrazoic sporocysts. The sporocysts (n = 10) measured 13 ± 0.5 µm × 11 ± 1.5 µm with an indistinct, irregularly placed, sporocyst residuum and usually contained 4 sporozoites, although an octozoic variant was observed. DNA was extracted from the snail kidney tissues and used as a template for polymerase chain reaction (PCR). PCR was used to determine the infection status of the snails; 13 of 36 (36%) specimens were found to be infected with a new Klossia species, and only 3 (8%) of these infections were detected using light microscopy. The complete nuclear 18S ribosomal DNA (1,800 bp) and mitochondrial genomes (6,775 bp) were generated, and they differentiated this parasite from the type species Klossia helicina and support the description of this new Klossia species, Klossia razorbacki n. sp. This is the first Klossia species to be described from any North American snail.
Herein we describe a new species and propose a new genus, Posthovitellinum psiloterminae n. gen., n. sp. (Lissorchiidae: Asymphylodorinae), based on specimens that infect the intestine of Cyclocheilos enoplos (Bleeker, 1849) (Cypriniformes: Cyprinidae), a migratory riverine carp from the Mekong River (Dong Thap province, Vietnam). The new species is assigned to Lissorchiidae by having a combination of features: spinous tegument, subterminal oral sucker, pre-equatorial ventral sucker, median and pretesticular ovary, submarginal genital pore at level of the ventral sucker, follicular vitellarium distributing in 2 lateral fields, and lacking eyespot pigment in the adult. It cannot be assigned to any existing asymphylodorine genus because it has the combination of a well-developed cirrus-sac, an unarmed ejaculatory duct and metraterm, a follicular vitellarium distributing in 2 lateral fields located between the posterior margin of the ventral sucker and the mid-level of the testis, and a sinistral, submarginal genital pore. The new species has an elongate, claviform cirrus-sac, a single, large, elongate-oval testis at the posterior extremity of the body, operculate eggs, and an I-shaped excretory bladder with secondary branches at the level of the testis and extending anteriad to the level of the pharynx. Bayesian inference analysis of the partial large subunit ribosomal DNA gene (28S rDNA) recovered the new species sister to Asaccotrema vietnamienseSokolov and Gordeev, 2019; these species differed by 118 nucleotides (12%; 983 bp fragment). This is the first lissorchiid reported from the Mekong River; only the second from southern Vietnam; and the fourth reported from a cyprinid fish in Vietnam. The aforementioned phylogenetic analysis included previously unpublished sequences representing lissorchiids infecting the intestine of North American suckers (Cypriniformes: Catostomidae): Lissorchis cf. nelsoni from spotted sucker; Minytrema melanops (Rafinesque, 1820) and Lissorchis cf. gullaris (immature) from smallmouth buffalo, Ictiobus bubalus (Rafinesque, 1818). Asymphylodora atherinopsidisAnnereaux, 1947, herein is treated as a species incertae sedis. The 28S tree topology suggests that Lissorchiinae may comprise more than 1 lineage, but additional species are needed to confidently assert this.
Adult specimens of Pomphorhynchus fuhaiensis were identified from common carp (Cyprinus carpio L.) in Ulungur Lake of northwest China, and acanthors, acanthellae, cystacanths dissected from Gammarus lacustris in a small tributary of Ulungur River for the first time. The acanthocephalans were also found in crucian carp (Carassius carassius L.), tench (Tinca tinca L.), oriental bream (Abramis brama orientalis Berg), and ide (Leuciscus idus L.) in the lake. This species is distinguished from other species in Pomphorhynchus by its large, spherical bulb and very long neck as well as by a cylindrical proboscis armed with 15–17 longitudinal rows of 9–12 hooks each. The anterior proboscis hooks are almost uniform in size and shape, the sixth hook in a short row and the seventh hook in long row decrease abruptly in size posteriorly with the last end hook being a little larger than the prebasal hook, and in a ring; posterior proboscis hooks much more widely spaced. Furthermore, the lemnisci are claviform. The mean neck:trunk ratio is about 0.5, which is larger than most other species in Pomphorhynchus. Females are larger than males. In males, the testes are in one-third to one-half of the trunk, equal, ovoid–spheroid, usually contiguous, and small relative to the body size, and there are 6 ovoid cement glands. Pomphorhynchus fuhaiensis is similar to Pomphorhynchus laevis but can be distinguished by the number of longitudinal rows of hooks. Pomphorhynchus laevis is armed with 18–20 longitudinal rows of 11–13 hooks, P. fuhaiensis is armed with 15–17 longitudinal rows of 9–12 hooks.
As presently recognized, Herpetodiplostomum is a small genus of proterodiplostomid digeneans parasitic in crocodilians. Most members of Cheloniodiplostomum, a genus of proterodiplostomids that parasitize chelonians, were originally placed within Herpetodiplostomum. The 2 genera were distinguished based on the presence/absence of papillae on the holdfast organ and anterior extent of vitellarium. Our study of Herpetodiplostomum and Cheloniodiplostomum revealed a lack of consistent morphological differences between the genera. Therefore, we consider Cheloniodiplostomum to be a junior synonym of Herpetodiplostomum. Recent molecular phylogenetic studies included a number of proterodiplostomid taxa; however, DNA sequence data are not available for any Herpetodiplostomum or Cheloniodiplostomum species. Herein, we describe a new Herpetodiplostomum species from Geoffroy's side-necked turtle Phrynops geoffroanus from Mato Grosso State, Brazil. The new species can be distinguished from other Herpetodiplostomum species based on the distribution of vitellarium, level of development of holdfast papillae, and ratio of prosoma:opisthosoma width, among other characters. We provide the first molecular phylogeny of the Proterodiplostomidae that includes a Herpetodiplostomum species. The limited geographic distribution of Herpetodiplostomum spp. within the geographic ranges of caimans suggests a secondary host switching event from crocodilians to chelonian definitive hosts in the evolution of Herpetodiplostomum.
The present study aimed to describe the natural trend of Eimeria spp. oocysts shedding in different breeds of adult goats raised in a sub-Alpine ecosystem by quantitative copromicroscopic analysis. A 1-yr long longitudinal study was planned in a goat herd composed of non-native (Alpine) and autochthonous (Nera di Verzasca) breeds. FLOTAC double technique was implemented to analyze individual fecal samples collected monthly from 2 groups (non-native and native) of 10 goats each. A generalized linear mixed model, in which each goat was included as a random intercept effect, identified 4 predictors of logarithmically transformed oocysts per gram shedding: the number of days from parturition (F = 27.744; P < 0.001), the interaction between the goat breed and the number of days from parturition (F = 8.407; P = 0.004), the interaction between the number of days from parturition and the number of parturitions (F = 6.371; P = 0.012), and the interaction between the monthly rainfalls and temperatures (F = 4.536; P = 0.023). These results indicate that the physiological status (number of days from parturition) of adult goats and its interaction with other individual features (number of parturitions and breed) can affect Eimeria spp. oocysts shedding. Besides, a combination of climatic variables can pose a risk for Eimeria spp. infection in goats reared in the sub-Alpine environment.
We investigated the effect of Schistosoma japonicum adenylate kinase 1 (Sjak1) on the growth and development of schistosomula. Quantitative real-time PCR showed that Sjak1 mRNA was expressed in 3-, 10-, 14-, 18-, and 21-day-old schistosomula, and its levels increased gradually with the development of S. japonicum. Using immunohistochemical techniques, ak1 protein was found to be mainly distributed in the tegument and some parenchymal tissues of the schistosomula. Double-stranded RNA-mediated knockdowns of ak1 decreased ak1 mRNA transcripts by more than 90%, and western blot results showed that expression of ak1 protein was decreased by 66%. Scanning electron microscopy following the RNA-mediated ak1 knockdown showed that the sensory papillae did not develop. Transmission electron microscopy showed a lower mean thickness of the tegument in the Sjak1 interference group than in the negative control group. Terminal deoxynucleotidyl transferase dUTP nick-end labeling suggested higher apoptosis in the interference group than the negative control group. These results showed that ak1 may be involved in the growth and development of S. japonicum schistosomula and especially in the development of the integument. Consequently, ak1 may be a potential target in developing prevention methods for schistosomiasis in the future.
We herein redescribe the enigmatic Cathariotrema selachii (MacCallum, 1916) Johnston and Tiegs, 1922 based on the holotype, paratypes, and newly collected specimens infecting the olfactory organ of 5 shark species from the Gulf of Mexico (all new host records): scalloped hammerhead shark, Sphyrna lewini (Griffith and Smith, 1834) (Carcharhiniformes: Sphyrnidae); great hammerhead shark, Sphyrna mokarran (Rüppell, 1837); blacktip shark, Carcharhinus limbatus (Müller and Henle, 1839) (Carcharhiniformes: Carcharhinidae); spinner shark, Carcharhinus brevipinna (Müller and Henle, 1839); and Atlantic sharpnose shark, Rhizoprionodon terraenovae (Richardson, 1836) (Carcharhinidae). These specimens were morphologically indistinguishable from each other and from MacCallum's holotype and paratypes. Those sequenced had identical first internal transcribed spacer (ITS1) and large subunit ribosomal DNA (28S) nucleotide sequences. As such, C. selachii infects sharks of 2 orders (Carcharhiniformes, Lamniformes) and 3 families (Carcharhinidae, Sphyrnidae, Lamnidae) in the Northwestern Atlantic Ocean (type locality) and Gulf of Mexico (new records herein). This report is the first of new specimens of C. selachii in the Atlantic Ocean Basin in 95 yr and corrects long-standing error cascades and ambiguities concerning the morphology and systematic placement of C. selachii. Considering morphology and nucleotide-based phylogenetic evidence (28S, Bayesian analysis), we herein emend monotypic CathariotremaJohnston and Tiegs, 1922 and propose Cathariotrematinae Bullard n. subfam. for it and 4 other genera (all formerly assigned to Merizocotylinae Johnston and Tiegs, 1922). These genera comprise species infecting only the nose of sharks (monotypic Cathariotrema, SqualotremaKearn and Green, 1983 and SeptitremaKheddam, Chisholm, and Tazerouti, 2020 plus 3 species of TriloculotremaKearn, 1993) and nose of a chimaera (monotypic HolocephalocotyleDerouiche, Neifar, Gey, Justine, and Tazerouti, 2019). Cathariotrematinae differs from Merizocotylinae by having a 3-part attachment organ and by lacking open loculi that symmetrically encircle a cluster of >2 loculi in the center of the haptor. Monophyletic Cathariotrematinae (with sequences representing species of Cathariotrema, Triloculotrema, and Holocephalocotyle only) was sister to monophyletic Merizocotylinae, which together were sister to monophyletic Calicotylinae Monticelli, 1903. These subfamilies comprise a monophyletic group of monocotylids that have a double vagina and infect extrabranchial, enclosed niches (urogenital system, body cavity, olfactory chamber/nose) on their shark, ray, and chimaera hosts (all other monocotylids have a single vagina and infect the gill or body surfaces of rays only). Monocotylinae Taschenberg, 1879 and Decacotylinae Chisholm, Wheeler, and Beverley-Burton, 1995 were recovered as monophyletic. Heterocotylinae Chisholm, Wheeler, and Beverley-Burton, 1995 remained paraphyletic. We accept ParacalicotyleSzidat, 1970.
The parasitic protist Trichomonas vaginalis is the causative agent of trichomoniasis, a highly prevalent sexually transmitted infection. The organism is known to accumulate substantial deposits of the polysaccharide glycogen, which is believed to serve as a store of carbon and energy that can be tapped during periods of nutrient limitation. Such nutrient limitation is likely to occur when T. vaginalis is transmitted between hosts, implying that glycogen may play an important role in the lifecycle of the parasite. Both T. vaginalis glycogen synthase and glycogen phosphorylase, key enzymes of glycogen synthesis and degradation, respectively, have been cloned and characterized, and neither enzyme is subject to the post-translational controls found in other, well-characterized eukaryotic systems. Thus, it is unclear how glycogen metabolism is regulated in this organism. Here we use a glucose limitation/re-feeding protocol to show that the activities of key enzymes of glycogen synthesis do not increase during re-feeding when glycogen synthesis is stimulated. Rather, a simple model appears to operate with glycogen storage being driven by the extracellular glucose concentration.
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.
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