Publications by authors named "Hong-Bin Yan"

46 Publications

Hydatigera taeniaeformis in urban rats (Rattus rattus) in Faisalabad, Pakistan.

Infect Genet Evol 2021 Apr 24;92:104873. Epub 2021 Apr 24.

State Key Laboratory of Veterinary Etiological Biology, National Professional Laboratory for Echinococcosis, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease, Yangzhou 225009, Jiangsu Province, PR China. Electronic address:

Hydatigera taeniaeformis formerly referred to as Taenia taeniaeformis is a cestode of cats (definitive hosts) and rodents (intermediate hosts). The prevalence of the metacestode larval stage has been reported in rodents in many parts of the world even though the genetic polymorphisms or intraspecies variation is still understudied. Here, we report a prevalence of 22.09% (38/172) from an urban rodent population in Pakistan and a nucleotide diversity (cox1) of 0.00463 among the population. Infection was higher in male (27.85%) and adult (32.29%) rats than female and sub-adult/young rats. Interestingly, The median-joining network and phylogenetic construction comprising isolates from China, Japan, Kenya, Laos, Malaysia, Senegal, the United Arab Emirates, and countries in Europe demonstrated that Pakistani H. taeniaeformis are closer to Asian and African population than those of European origin. The results of the study will add-in preliminary data for H. taeniaeformis and will also contribute to understand the global molecular epidemiology and population structure of H. taeniaeformis.
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http://dx.doi.org/10.1016/j.meegid.2021.104873DOI Listing
April 2021

Genetic Evolution and Implications of the Mitochondrial Genomes of Two Newly Identified spp. in Rodents From Qinghai-Tibet Plateau.

Front Microbiol 2021 23;12:647119. Epub 2021 Mar 23.

State Key Laboratory of Veterinary Etiological Biology, National Professional Laboratory for Animal Echinococcosis, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.

The larva of Taeniidae species can infect a wide range of mammals, causing major public health and food safety hazards worldwide. The Qinghai-Tibet Plateau (QTP), a biodiversity hotspot, is home to many species of rodents, which act as the critical intermediate hosts of many Taeniidae species. In this study, we identified two new larvae of spp., named and , collected from the plateau pika () and the Qinghai vole (), respectively, in QTP, and their mitochondrial genomes were sequenced and annotated. Phylogenetic trees based on the mitochondrial genome showed that has the closest genetic relationship with , while was contained in a monophyletic group with , , and . Biogeographic scenarios analysis based on split time speculated that the speciation of (∼5.49 Mya) is due to host switching caused by the evolution of its intermediate host. Although the reason for (∼13.11 Mya) speciation is not clear, the analysis suggests that it should be infective to a variety of other rodents following the evolutionary divergence time of its intermediate host and the range of intermediate hosts of its genetically close species. This study confirms the species diversity of Taeniidae in the QTP, and speculates that the uplift of the QTP has not only a profound impact on the biodiversity of plants and animals, but also that of parasites.
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http://dx.doi.org/10.3389/fmicb.2021.647119DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8021716PMC
March 2021

in Qinghai-Tibet plateau: population structure and confirmation of additional endemic areas.

Parasitology 2021 Mar 24:1-8. Epub 2021 Mar 24.

State Key Laboratory of Veterinary Etiological Biology/National Professional Laboratory for Animal Echinococcosis/Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, CAAS, Lanzhou730046, Gansu Province, People's Republic of China.

Echinococcus shiquicus is currently limited to the Qinghai–Tibet plateau, a large mountainous region in China. Although the zoonotic potential remains unknown, progress is being made on the distribution and intermediate host range. In this study, we report E. shiquicus within Gansu and Qinghai provinces in regions located not only around the central areas but also the southeast edge of the plateau and describe their genetic relationship with previous isolates from the plateau. From 1879 plateau pikas examined, 2.39% (95% CI 1.79–3.18) were infected with E. shiquicus. The highest prevalence of 10.26% (4.06–23.58) was recorded in Makehe town, Qinghai province. Overall the prevalence was marginally higher in Qinghai (2.5%, CI 1.82–3.43) than in Gansu (2%, CI 1.02–3.89). The cox1 and nad1 genes demonstrated high and low haplotype and nucleotide diversities, respectively. The median-joining network constructed by the cox1–nad1 gene sequences demonstrated a star-like configuration with a median vector (unsampled haplotype) occupying the centre of the network. No peculiar distinction or common haplotype was observed in isolates originating from the different provinces. The presence of E. shiquicus in regions of the southeast and northeast edges of the Qinghai–Tibet plateau and high genetic variation warrants more investigation into the haplotype distribution and genetic polymorphism by exploring more informative DNA regions of the mitochondrial genome to provide epidemiologically useful insight into the population structure of E. shiquicus across the plateau and its axis.
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http://dx.doi.org/10.1017/S0031182021000512DOI Listing
March 2021

First Report on Molecular Characterization of Isolates From Sheep and Goats in Faisalabad, Pakistan.

Front Vet Sci 2020 10;7:594599. Epub 2020 Nov 10.

State Key Laboratory of Veterinary Etiological Biology, National Professional Laboratory for Animal Echinococcosis, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.

is the larval stage of commonly found in the brain (cerebral form), intramuscular and subcutaneous tissues (non-cerebral form) of ungulates. Globally, few reports exist on the molecular characterization and genetic diversity of . with none available for Pakistan. The current study molecularly characterized 12 . isolates surgically recovered from sheep ( = 4) and goats ( = 8) from a total of 3,040 small ruminants using a portion of the cytochrome oxidase subunit 1 (1) mitochondrial () gene. NCBI BLAST search confirmed the identity of each isolate. A high haplotype and a low nucleotide diversity with three haplotypes from the 12 isolates were observed. The findings suggest the existence of unique haplotypes of in Pakistan. The negative value of Tajima's and the positive value of Fu's Fs were inconsistent with population expansion, however, the sample size was small. Bayesian phylogeny revealed that all Pakistani isolates alongside the Chinese sequences (obtained from GenBank) constituted a cluster while sequences from other regions constituted another cluster. This is the first molecular study to determine the genetic diversity of . in Pakistan and serves as a foundation for prospective studies on the prevalence and population structure of in the country. Furthermore, in this study, we amplified only a partial segment of the 1 gene from a limited sample size. This could have implications on the interpretation of the actual population structure in reality. Thus, we recommend future studies to consider a larger sample size in a massive epidemiological survey for further insights.
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http://dx.doi.org/10.3389/fvets.2020.594599DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7683608PMC
November 2020

First report on the phylogenetic relationship, genetic variation of Echinococcus shiquicus isolates in Tibet Autonomous Region, China.

Parasit Vectors 2020 Nov 23;13(1):590. Epub 2020 Nov 23.

State Key Laboratory of Veterinary Etiological Biology/National Professional Laboratory for Animal Echinococcosis/Key Laboratory of Veterinary Parasitology of Gansu Province/Key Laboratory of Zoonoses of Agriculture Ministry, Lanzhou Veterinary Research Institute (CAAS), Lanzhou, 730046, Gansu, People's Republic of China.

Background: Cystic or alveolar echinococcosis caused by the larval stages of Echinococcus spp. is a very severe zoonotic helminth infection. Echinococcus shiquicus is a newly discovered species that has only been reported in the Qinghai and Sichuan provinces of the Qinghai-Tibet plateau, China where, to date, it has only been confirmed in Tibetan foxes and wild small mammal populations of the Tibetan plateau. Information on its genetic and evolutionary diversity is scanty. The aim of this study was to investigate the prevalence of E. shiquicus in plateau pikas (Ochotona curzoniae), a known intermediate host, and to determine the genetic variation and phylogenetic relationship of the E. shiquicus population in the Tibet region of China based on mitochondrial DNA.

Methods: Echinococcus shiquicus samples were collected from Damxung and Nyêmo counties (located in Tibet Autonomous Region, China). The mitochondrial cox1 and nad1 gene sequences were analyzed, and the genetic diversity and epidemiology of E. shiquicus in the region were discussed based on the results.

Results: The prevalence of E. shiquicus in pikas in Damxung and Nyêmo counties was 3.95% (6/152) and 6.98% (9/129), respectively. In combination with previous public sequence data, the haplotype analysis revealed 12 haplotypes (H) characterized by two distinct clusters (I and II), and a sequence distance of 99.1-99.9% from the reference haplotype (H1). The diversity and neutrality indices for the entire E. shiquicus populations were: haplotype diversity (Hd) ± standard deviation (SD) 0.862 ± 0.035; nucleotide diversity (Hd ± SD) 0.0056 ± 0.0003; Tajima's D 0.876 (P > 0.05); and Fu's F 6.000 (P > 0.05).

Conclusions: This was the first analysis of the newly discovered E. shiquicus in plateau pikas in the Tibet Autonomous Region of China. The neutrality indices suggest a deficiency of alleles, indicative of a recent population bottleneck.
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http://dx.doi.org/10.1186/s13071-020-04456-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7686673PMC
November 2020

In-House Developed ELISA Indicates High Prevalence of Anti- IgG in Sheep Population-An Update from Pakistan.

Pathogens 2020 Oct 29;9(11). Epub 2020 Oct 29.

State Key Laboratory of Veterinary Etiological Biology, National Professional Laboratory of Animal Hydatidosis, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.

Cystic echinococcosis (CE) is a World Health Organization (WHO)-listed neglected tropical farm economy jeopardizing and public health concern disease. This study was aimed at furnishing sero-epidemiological baseline data of CE in sheep in Pakistan, where data are non-existent. For this purpose, two sheep-rich provinces of Pakistan were selected, and 728 sheep sera were collected using probability proportional to size (PPS) statistical technique. Epidemiological information was recorded on a questionnaire for the estimation of potential risk factors. The serum samples were analyzed for IgG antibodies against using an in-house-developed EgAgB-based ELISA kit. The overall seroprevalence recorded was 21.98% (160/728) in the tested sheep, suggesting higher seropositivity in sheep from Punjab (23.73%) as compared to Khyber Pakhtunkhwa (KPK) (19.04%). The overall apparent prevalence observed by this ELISA method was almost similar to the calculated true prevalence (21.77%). Prevalence was significantly different ( < 0.05) among sheep from different districts. Higher prevalence was found in females (22.54%, OR 1.41), age group > 5 years (29.66%, OR 1.64), crossbreeds (42.85%, OR 2.70), and sheep with pasture access (25.96%, OR 3.06). Being in age group > 5 years and having pasture access were the factors significantly associated with seropositivity ( < 0.05). This study provides serological evidence of . infection in sheep and can be used as a model for screening of the sheep globally.
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http://dx.doi.org/10.3390/pathogens9110905DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7693474PMC
October 2020

A new molecular nomenclature for : mitochondrial DNA sequences reveal sufficient diversity suggesting the assignment of major haplotype divisions.

Parasitology 2021 Mar 23;148(3):311-326. Epub 2020 Oct 23.

State Key Laboratory of Veterinary Etiological Biology/National Professional Laboratory for Animal Echinococcosis/Key Laboratory of Veterinary Parasitology of Gansu Province/Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou730046, Gansu Province, P. R. China.

Cysticercosis caused by the metacestode larval stage of Taenia hydatigena formerly referred to as Cysticercus tenuicollis is a disease of veterinary importance that constitutes a significant threat to livestock production worldwide, especially in endemic regions due to condemnation of visceral organs and mortality rate of infected young animals. While the genetic diversity among parasites is found to be potentially useful in many areas of research including molecular diagnostics, epidemiology and control, that of T. hydatigena across the globe remains poorly understood. In this study, analysis of the mitochondrial DNA (mtDNA) of adult worms and larval stages of T. hydatigena isolated from dogs, sheep and a wild boar in China showed that the population structure consists of two major haplogroups with very high nucleotide substitutions involving synonymous and non-synonymous changes. Compared with other cestodes such as Echinococcus spp., the genetic variation observed between the haplogroups is sufficient for the assignment of major haplotype or genotype division as both groups showed a total of 166 point-mutation differences between the 12 mitochondrial protein-coding gene sequences. Preliminary analysis of a nuclear protein-coding gene (pepck) did not reveal any peculiar changes between both groups which suggests that these variants may only differ in their mitochondrial makeup.
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http://dx.doi.org/10.1017/S003118202000205XDOI Listing
March 2021

Thioredoxin Peroxidase 2 Regulates Protective Th2 Immune Response in Mice by Directly Inducing Alternatively Activated Macrophages.

Front Immunol 2020 25;11:2015. Epub 2020 Sep 25.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.

infection can induce macrophages into the alternatively activated phenotype, which is primarily associated with the development of a polarized Th2 immune response. In the present study, we examined the immunomodulatory effect of thioredoxin peroxidase-2 (TsTPX2), a protein derived from ES products, in the regulation of Th2 response through direct activation of macrophages. The location of TsTPX2 was detected by immunohistochemistry and immunofluorescence analyses. The immune response induced by rTsTPX2 was characterized by analyzing the Th2 cytokines and Th1 cytokines in the peripheral blood. The rTsTPX2-activated macrophages (M) were tested for polarization, their ability to evoke naïve CD4 T cells, and resistance to the larval infection after adoptive transfer in BALB/c mice. The immunolocalization analysis showed TsTPX2 in cuticles and stichosome of ML. The immunostaining was detected in cuticles and stichosome of Ad3 and ML, as well as in tissue-dwellings around ML after the intestines and muscle tissues of infected mice were incubated with anti-rTsTPX2 antibody. Immunization of BALB/c mice with rTsTPX2 could induce a Th1-suppressing mixed immune response given the increased levels of Th2 cytokines (IL-4 and IL-10) production along with the decreased levels of Th1 cytokines (IFN-γ, IL-12, and TNF-α). studies showed that rTsTPX2 could directly drive RAW264.7 and peritoneal macrophages to the M2 phenotype. Moreover, M could promote CD4 T cells polarized into Th2 type . Adoptive transfer of M into mice suppressed Th1 responses by enhancing Th2 responses and exhibited a 44.7% reduction in adult worm burden following challenge with infective larval, suggesting that the TsTPX2 is a potential vaccine candidate against trichinosis. Our study showed that TsTPX2 would be at least one of the molecules to switch macrophages into the M2 phenotype during infection, which provides a new therapeutic approach to various inflammatory disorders like allergies or autoimmune diseases.
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http://dx.doi.org/10.3389/fimmu.2020.02015DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7544948PMC
September 2020

Preliminary information on the prevalence and molecular description of Taenia hydatigena isolates in Pakistan based on mitochondrial cox1 gene.

Infect Genet Evol 2020 11 28;85:104481. Epub 2020 Jul 28.

State Key Laboratory of Veterinary Etiological Biology, National Professional Laboratory for Echinococcosis, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, PR China. Electronic address:

Taenia hydatigena is a cestode of veterinary importance. Infection with the metacestode larval stage results in cysticercosis, which poses a serious challenge to the livestock industry worldwide. Globally, there are numerous reports on cysticercosis caused by T. hydatigena in sheep and goat but a lack of data on the prevalence and genetic diversity exists for Pakistan. We designed this study to provide an insight into the disease status as well as investigate the genetic variation among the recovered isolates based on the mitochondrial cox1 gene. In this study, we examined small ruminants (sheep and goats) slaughtered in Faisalabad in eastern Punjab province of Pakistan for T. hydatigena metacestodes and described the population structure and genetic variation using the cytochrome c oxidase subunit 1 (cox1) mitochondrial gene. Overall, a prevalence of 4.40% (goat =4.67% sheep = 4.07%) from a total of 2225 small ruminant carcasses (sheep = 983, goats = 1242) was observed. Based on the NCBI BLAST search and Bayesian phylogeny, the identity of all isolates was confirmed via their nucleotide sequences. The diversity indices indicated a high haplotype and a low nucleotide diversity with 43 haplotypes from 98 isolates. The results also show the existence of unique haplotypes of T. hydatigena in Pakistan as demonstrated by the significant negative values of Tajima's D and Fu's Fs neutrality test suggesting a recent population expansion. The median-joining network of the partial cox1 sequence dataset showed the existence of two main haplotypes detected in both sheep and goat populations. This study shows that the prevalence of cycticercosis due to T. hydatigena is below 5% in sheep and goats in Faisalabad, Punjab, Pakistan. The molecular analysis of the partial cox1 gene also indicates a high degree of genetic variation with the existence of rare haplotypes. These findings represent a preliminary report on the prevalence and genetic variation of T. hydatigena in Pakistan and serve as baseline information for future studies on the prevalence and population structure of T. hydatigena in the country.
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http://dx.doi.org/10.1016/j.meegid.2020.104481DOI Listing
November 2020

Tritiated Water Induces Toxicity in Human Umbilical Vein Vascular Endothelial Cells via IL8.

Dose Response 2020 Jul-Sep;18(3):1559325820938541. Epub 2020 Jul 7.

State Key Laboratory of Radiation Medicine and Protection, School of Radiation Medicine and Protection, Soochow University, Suzhou, People's Republic of China.

We aimed to determine the toxic effects of tritiated water (HTO) on 12 generations (T1-T12) of human umbilical vein vascular endothelial cells (HUVECs) and elucidate the underlying mechanisms. We evaluated cellular senescence, interleukin (IL) 8 concentrations, and angiogenesis using β-galactosidase staining, enzyme-linked immunosorbent assay, and in vitro assays, respectively. The adhesion properties of contaminated cells and differentially expressed genes were assessed using the xCELLigence RTCA SP system and gene chip analysis, respectively. We found that long-term exposure to low levels of HTO can reduce the adhesion of HUVECs to the cellular matrix as well as their angiogenic capacity, while increasing their permeability, senescence, and adhesion to monocytes. Interleukin 8 activated the p38 and Epidermal Growth Factor Receptor (EGFR) pathways in HTO-treated cells and hence was identified as a key candidate of biomarker. The present study clarified the toxicity of HTO in vascular endothelial cells and identified IL8 as a novel protective target with important theoretical and practical values.
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http://dx.doi.org/10.1177/1559325820938541DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7343372PMC
July 2020

Echinococcus granulosus (sensu stricto) (G1, G3) and E. ortleppi (G5) in Pakistan: phylogeny, genetic diversity and population structural analysis based on mitochondrial DNA.

Parasit Vectors 2020 Jul 13;13(1):347. Epub 2020 Jul 13.

State Key Laboratory of Veterinary Etiological Biology, National Professional Laboratory of Animal Hydatidosis, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, Gansu, People's Republic of China.

Background: Cystic echinococcosis (CE) is a serious tapeworm infection caused by Echinococcus granulosus (sensu lato) which infects a wide range of animals and humans worldwide. Despite the millions of livestock heads reared in Pakistan, only a few reports on CE prevalence and even fewer on the genetic diversity are available for the country. Meanwhile, the available reports on the genetic diversity are predominantly based on short sequences of the cox1 gene.

Methods: To close this knowledge gap, this study was designed to investigate the genetic diversity and population structure of Echinococcus spp. in Pakistan using the complete mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes.

Results: Based on BLAST searches of the generated cox1 and nad1 gene sequences from a total of 60 hydatid cysts collected from cattle (n = 40) and buffalo (n = 20), 52 isolates were identified as E. granulosus (s.s.) (G1, G3) and 8 as E. ortleppi (G5). The detection of the G5 genotype represents the first in Pakistan. The phylogeny inferred by the Bayesian method using nucleotide sequences of cox1-nad1 further confirmed their identity. The diversity indices indicated a high haplotype diversity and a low nucleotide diversity. The negative values of Tajima's D and Fu's Fs test demonstrated deviation from neutrality suggesting a recent population expansion.

Conclusions: To the best of our knowledge, this report described the genetic variation of E. granulosus population for the first time in Pakistan using the complete cox1 and nad1 mitochondrial genes and confirms E. ortleppi as one of the causative agents of CE among livestock in Pakistan. While this report will contribute to baseline information for CE control, more studies considering species diversity and distribution in different hosts across unstudied regions of Pakistan are highly needed.
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http://dx.doi.org/10.1186/s13071-020-04199-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7359271PMC
July 2020

Complete mitochondrial genome analysis confirms the presence of Echinococcus granulosus sensu lato genotype G6 in Nigeria.

Infect Genet Evol 2020 10 26;84:104377. Epub 2020 May 26.

State Key Laboratory of Veterinary Etiological Biology/National Professional Laboratory of Animal Hydatidosis/Key Laboratory of Veterinary Parasitology of Gansu Province/Lanzhou Veterinary Research Institute, CAAS, Lanzhou 730046, PR China. Electronic address:

Cystic echinococcosis (CE) is common in Africa especially in northern and eastern countries where it is highly prevalent and mainly caused by Echinococcus granulosus sensu stricto and Echinococcus granulosus sensu lato (G6/7). In a recent epidemiological survey in Nigeria, the G6/7 genotype was reportedly responsible for the majority of CE infection. In this study, considering the taxonomic challenges of the G6/7 genotype and the limitation of the cox1/nad1 genes in resolving the differences, we sequenced the complete mitochondrial (mt) genome of seven larval isolates of E. granulosus s.l. G6/7 haplotypes recently reported in Nigeria to correctly assign them to either (G6/7) genotype and to understand the mt genome variation with isolates from other geographical regions. On analysis, a total of 13,731 bp in a covalently closed circular molecule were realized. The total mt length was ≥10 bp than previously reported G6 and G7 mt genome sequences. Also, the median-joining network and phylogeny based on the 12 protein-coding sequences correctly identified them as G6 genotype. Since longer mt genome sequences have shown some advantage over individual genes in resolving taxonomic challenges, we confirm that the genotype responsible for the majority of CE infection in livestock in Nigeria is the G6 genotype and the availability of the complete mt genome from different Nigerian intermediate hosts will prove useful in future genetic population studies across the country and the West African sub-region.
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http://dx.doi.org/10.1016/j.meegid.2020.104377DOI Listing
October 2020

Prevalence and distribution of Echinococcus spp. in wild and domestic animals across Africa: A systematic review and meta-analysis.

Transbound Emerg Dis 2020 Nov 15;67(6):2345-2364. Epub 2020 May 15.

State Key Laboratory of Veterinary Etiological Biology/National Professional Laboratory for Animal Echinococcosis/Key Laboratory of Veterinary Parasitology of Gansu Province/Key Laboratory of Zoonoses of Agriculture Ministry, Lanzhou Veterinary Research Institute, CAAS, Lanzhou, P. R. China.

Cystic echinococcosis (CE), a worldwide zoonosis, is highly prevalent in Africa particularly in northern and eastern Africa where data are more abundant than other regions. However, harmonization of available data through systematic review and meta-analysis may foster improved transboundary cooperation for the control of CE in Africa. Using the Preferred Reporting Items for Systematic Reviews and Meta-analyses guidelines, research articles (from 2000 to 2019) were retrieved from ScienceDirect, PubMed, African Journals OnLine and Google Scholar databases. A total of 98 studies of 806,624 animals from 13 countries comprising 264,016 goats, 247,326 sheep, 251,106 cattle, 28,314 camels, 4,764 buffaloes, 2,920 equids, 1,966 pigs, 408 wild boars and 50 Norway rats were available for systematic review and meta-analysis of pooled prevalence including 5,048 dogs, 345 lions, 220 hyenas, 94 wolves and 47 jackals/foxes analysed for Echinococcus infection. In total, 46,869 animals were infected and pooled prevalence of CE in intermediate hosts was highest in camels (17.1%; 95% CI: 12.1-22.8) and lowest in pigs (0.3%; 95% CI: 0.1-0.6). Results also showed uneven species/genotype distribution across the continent such that Echinococcus granulosus sensu stricto (G1, G3) constituted 74.45% of the total isolates from East Africa, E. canadensis (G6/7) accounted for 60.3% and 97.4% in North and West Africa, respectively, while 81.3% of E. ortleppi (G5) were recorded for southern Africa. The comparatively higher prevalence estimates for eastern and northern Africa than other regions indicate where efforts on CE management should now be given greater attention in Africa. Additionally, this study also advocates for better cooperation between countries within the same sub-region and the establishment of joint CE control programmes.
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http://dx.doi.org/10.1111/tbed.13571DOI Listing
November 2020

Molecular Identification of Taenia hydatigena from Sheep in Khartoum, Sudan.

Korean J Parasitol 2020 Feb 29;58(1):93-97. Epub 2020 Feb 29.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, P.R. China.

The cestode Taenia hydatigena uses canids, primarily dogs, as definitive hosts, while the metacestode larval stage cysticercus infects a range of intermediate hosts, including domestic animals such as goats, sheep, and pigs. Cysticercosis due to T. hydatigena has large veterinary and economic drawbacks. Like other taeniids, e.g., Echinococcus, intraspecific variation is found among the members of the genus Taenia. In Africa, few studies are available on the epidemiology and distribution of T. hydatigena, and even fewer studies are available on its genetic variation. In this study, we molecularly identified 11 cysticerci from sheep in Sudan and demonstrated the genetic variation based on the NADH dehydrogenase subunit 1 (nad1) and cytochrome c oxidase subunit 1 (cox1) mitochondrial genes. The isolates were correctly identified as T. hydatigena with more than 99% similarity to those in the GenBank database. Low diversity indices and insignificant neutrality indices were observed, with 3 and 2 haplotypes for the nad1 and cox1 genes, respectively. The results suggest the presence of unique T. hydatigena haplotypes in Sudan, as haplotypes with 100% similarity were not found in the GenBank database. With few available studies on the genetic variation of T. hydatigena in Africa, this report represents the first insights into the genetic variation of T. hydatigena in Sudan and constitutes useful data.
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http://dx.doi.org/10.3347/kjp.2020.58.1.93DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7066435PMC
February 2020

Development of genome-wide polymorphic microsatellite markers for Trichinella spiralis.

Parasit Vectors 2020 Feb 11;13(1):58. Epub 2020 Feb 11.

Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun, 130062, Jilin, People's Republic of China.

Background: Trichinella nematodes are globally distributed food-borne pathogens, in which Trichinella spiralis is the most common species in China. Microsatellites are a powerful tool in population genetics and phylogeographic analysis. However, only a few microsatellite markers were reported in T. spiralis. Thus, there is a need to develop and validate genome-wide microsatellite markers for T. spiralis.

Methods: Microsatellites were selected from shotgun genomic sequences using MIcroSAtellite identification tool (MISA). The identified markers were validated in 12 isolates of T. spiralis in China.

Results: A total of 93,140 microsatellites were identified by MISA from 9267 contigs in T. spiralis genome sequences, in which 16 polymorphic loci were selected for validation by PCR with single larvae from 12 isolates of T. spiralis in China. There were 7-19 alleles per locus (average 11.25 alleles per locus). The observed heterozygosity (H) and expected heterozygosity (H) ranged from 0.325 to 0.750 and 0.737 to 0.918, respectively. The polymorphism information content (PIC) ranged from 0.719 to 0.978 (average 0.826). Among the 16 loci, markers for 10 loci could be amplified from all 12 international standard strains of Trichinella spp.

Conclusions: Sixteen highly polymorphic markers were selected and validated for T. spiralis. Primary phylogenetic analysis showed that these markers might serve as a useful tool for genetic studies of Trichinella parasites.
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http://dx.doi.org/10.1186/s13071-020-3929-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7014596PMC
February 2020

Genetic variation of Echinococcus spp. in yaks and sheep in the Tibet Autonomous Region of China based on mitochondrial DNA.

Parasit Vectors 2019 Dec 27;12(1):608. Epub 2019 Dec 27.

State Key Laboratory of Veterinary Etiological Biology/National Professional Laboratory of Animal Hydatidosis, Key Laboratory of Veterinary Parasitology of Gansu Province/Lanzhou Veterinary Research Institute, CAAS, Lanzhou, 730046, Gansu, People's Republic of China.

Background: Cystic echinococcosis (CE) in humans and livestock is caused by Echinococcus granulosus (sensu lato). In China where CE is endemic, a number of studies have shown that Echinococcus granulosus (sensu stricto) is majorly responsible for CE. However, E. canadensis (G6) which is the second leading cause of CE is now being detected in most parts of the country. In this study, the species diversity and genetic variation of Echinococcus granulosus (s.l.) in four counties in Tibet Autonomous Region of China were investigated.

Methods: Infection with Echinococcus granulosus (s.s.) in yaks and sheep was identified using NADH dehydrogenase subunit 1 and 5 (nad1 and nad5) mitochondrial genes while the genotype G6 of E. canadensis initially diagnosed with NADH dehydrogenase subunit 1 (nad1) was further confirmed by analysis of the complete mitochondrial genome and a phylogenetic network constructed based on the nad2 and nad5 genes.

Results: Out of 85 hydatid cyst samples collected from slaughtered sheep (n = 54) and yaks (n = 31), 83 were identified as E. granulosus (s.s.) G1 (n = 77), G3 (n = 6) and 2 were identified as E. canadensis G6. Analysis of the nad1/nad5 genes revealed 16/17 mutations with 9/14 parsimony informative sites resulting in 15/14 haplotypes, respectively. Haplotype diversity (Hd) and nucleotide diversity (π) of E. granulosus (s.s.) population were 0.650 and 0.00127 for nad1 and 0.782 and 0.00306 for nad5, respectively, with an overall negative Tajima's D and Fu's Fs. A low F indicated no genetic difference between isolates from sheep and yaks.

Conclusion: Pockets of infection with E. canadensis (G6, G7, G8 and G10) have been previously reported in sheep, goats, yaks and/or humans in different parts of China. While the G6 genotype has been previously reported in sheep in the Tibet Autonomous Region, the detection in a yak in the present study represents the first to the best of our knowledge. Therefore, we recommend future surveys and control efforts to comprehensively investigate other potential intermediate hosts for the prevalence and genetic diversity of the E. canadensis group (G6, G7, G8 and G10) across the country and their inclusion into the existing CE control programme.
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http://dx.doi.org/10.1186/s13071-019-3857-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6935104PMC
December 2019

Correction to: First molecular description, phylogeny and genetic variation of Taenia hydatigena from Nigerian sheep and goats based on three mitochondrial genes.

Parasit Vectors 2019 11 21;12(1):547. Epub 2019 Nov 21.

State Key Laboratory of Veterinary Etiological Biology/National Professional Laboratory of Animal Hydatidosis/Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, People's Republic of China.

Following publication of the original article [1], the have authors flagged that the information in the legend of Fig. 1 is detailed in the wrong order.
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http://dx.doi.org/10.1186/s13071-019-3807-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6873671PMC
November 2019

First molecular description, phylogeny and genetic variation of Taenia hydatigena from Nigerian sheep and goats based on three mitochondrial genes.

Parasit Vectors 2019 Nov 5;12(1):520. Epub 2019 Nov 5.

State Key Laboratory of Veterinary Etiological Biology/National Professional Laboratory of Animal Hydatidosis/Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, People's Republic of China.

Background: Cysticercosis caused by the metacestode larval stage of Taenia hydatigena is a disease of veterinary and economic importance. A considerable level of genetic variation among isolates of different intermediate hosts and locations has been documented. Generally, data on the genetic population structure of T. hydatigena is scanty and lacking in Nigeria. Meanwhile, similar findings in other cestodes like Echinococcus spp. have been found to be of epidemiological importance. Our aim, therefore, was to characterize and compare the genetic diversity of T. hydatigena population in Nigeria based on three mitochondrial DNA markers as well as to assess the phylogenetic relationship with populations from other geographical regions.

Methods: In the present study, we described the genetic variation and diversity of T. hydatigena isolates from Nigerian sheep and goats using three full-length mitochondrial genes: the cytochrome c oxidase subunit 1 (cox1), NADH dehydrogenase subunit 1 (nad1), and NADH dehydrogenase subunit 5 (nad5).

Results: The median-joining network of concatenated cox1-nad1-nad5 sequences indicated that T. hydatigena metacestodes of sheep origin were genetically distinct from those obtained in goats and this was supported by high F values of nad1, cox1, and concatenated cox1-nad1-nad5 sequences. Genetic variation was also found to be higher in isolates from goats than from sheep.

Conclusions: To the best of our knowledge, the present study described the genetic variation of T. hydatigena population for the first time in Nigeria using full-length mitochondrial genes and suggests the existence of host-specific variants. The population indices of the different DNA markers suggest that analysis of long mitochondrial DNA fragments may provide more information on the molecular ecology of T. hydatigena. We recommend that future studies employ long mitochondrial DNA sequence in order to provide reliable data that would explain the extent of genetic variation in different hosts/locations and the biological and epidemiological significance.
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http://dx.doi.org/10.1186/s13071-019-3780-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6833231PMC
November 2019

A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections.

BMC Infect Dis 2019 Oct 16;19(1):854. Epub 2019 Oct 16.

State Key Laboratory of Veterinary Etiological Biology/ Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute, CAAS, Lanzhou, 730046, Gansu Province, People's Republic of China.

Background: Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum are four common large and medium-sized tapeworms parasitizing the small intestine of dogs and other canids. These parasites cause serious impact on the health and development of livestock. However, there are, so far, no commercially available molecular diagnostic kits capable of simultaneously detecting all four parasites in dogs. The aim of the study was therefore to develop a multiplex PCR assay that will accurately detect all four cestode infections in one reaction.

Methods: Specific primers for a multiplex PCR were designed based on corresponding mitochondrial genome sequences, and its detection limit was assessed by serial dilutions of the genomic DNAs of tapeworms examined. Furthermore, field samples of dog feces were tested using the developed assay.

Results: A multiplex polymerase chain reaction (PCR) assay was developed based on mitochondrial DNA (mtDNA) that accurately and simultaneously identify four cestode species in one reaction using specific fragment sizes of 592, 385, 283, and 190 bp for T. hydatigena, T. multiceps, T. pisiformis, and D. caninum, respectively. The lowest DNA concentration detected was 1 ng for T. hydatigena, T. multiceps and T. pisiformis, and 0.1 ng for D. caninum in a 25 μl reaction system. This assay offers high potential for the rapid detection of these four tapeworms in host feces simultaneously.

Conclusions: This study provides an efficient tool for the simultaneous detection of T. hydatigena, T. multiceps, T. pisiformis, and D. caninum. The assay will be potentially useful in epidemiological studies, diagnosis, and treatment of these four cestodes infections during prevention and control program.
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http://dx.doi.org/10.1186/s12879-019-4512-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6796438PMC
October 2019

Review of Cystic Echinococcosis in Nigeria: A Story of Neglect.

Acta Parasitol 2020 Mar 24;65(1):1-10. Epub 2019 Sep 24.

State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veterinary Parasitology of Gansu Province/Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, Gansu Province, People's Republic of China.

Purpose: Cystic echinococcosis (CE) caused by Echinococcus granulosus sensu lato is a widespread zoonotic disease of global concern. In Nigeria, the exact picture/status of CE is unclear, as most of the states are largely uninvestigated. Yet, as with every parasitic zoonosis, the first step towards planning a comprehensive management and control programme involves assessment of available national/regional prevalence data, host range, and risk factors at play in the transmission dynamics.

Methods: Published articles on echinococcosis were searched on PubMed and Africa Journal Online (AJOL) databases. Inclusion criteria were based on studies reporting prevalence of echinococcosis in animals and humans (including case reports) from 1970 to 2018.

Results: In this study, we evaluated and summarized cystic echinococcosis reports in Nigeria and found that post 1970-80s, studies on cystic echinococcosis have remained sparse regardless of the high prevalence recorded in the early years of CE investigation. In addition, information on the genetic population structure and the role of wildlife in CE transmission is still lacking.

Conclusions: This study appraises the prevalence and distribution of CE in Nigeria and identified areas where surveillance and control efforts should be focused and intensified.
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http://dx.doi.org/10.2478/s11686-019-00124-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7223537PMC
March 2020

Cystic echinococcosis in Nigeria: first insight into the genotypes of Echinococcus granulosus in animals.

Parasit Vectors 2019 Aug 7;12(1):392. Epub 2019 Aug 7.

State Key Laboratory of Veterinary Etiological Biology/National Professional Laboratory of Animal Hydatidosis, Lanzhou Veterinary Research Institute, CAAS, Lanzhou, 730046, Gansu, P. R. China.

Background: Cystic echinococcosis (CE) is a zoonosis caused by cestodes of Echinococcus granulosus (sensu lato) complex. In Nigeria, reports on the prevalence of CE, although limited, have been found to vary with location and host with higher prevalence and fertility rate observed in camels than other livestock. Until now, information regarding the molecular characteristics, genetic population structure, and genotypes of Echinococcus is lacking. Therefore, this study was aimed at addressing these gaps in knowledge.

Methods: We describe the genetic status of 31 Echinococcus isolates collected from slaughtered livestock (camels, cattle and goats) based on the full-length mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes.

Results: The resulting nucleotide sequences via the NCBI BLAST algorithm and Bayesian phylogeny of cox1 and cox1-nad1 genes using MrBayes v.3.1.2 showed that all isolates were clearly E. canadensis (G6/G7) and were 99-100% identical to previously reported G6/G7 haplotypes across Europe, Asia, North and East Africa.

Conclusions: Although, the G1 genotype is believed to be responsible for the majority of global CE burden, reports from a number of West African countries including Nigeria suggest that E. canadensis G6/G7 genotype could be the major causative agent of CE in the subregion. This study provides for the first time insight into the genetic population structure of Echinococcus species as well as implications for CE control in Nigeria.
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http://dx.doi.org/10.1186/s13071-019-3644-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6686243PMC
August 2019

Complete mitochondrial genomes confirm the generic placement of the plateau vole, .

Biosci Rep 2019 08 9;39(8). Epub 2019 Aug 9.

State Key Laboratory of Veterinary Etiological Biology/ National Professional Laboratory of Animal Hydatidosis/ Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute, CAAS, Lanzhou 730046, P.R. China

The plateau vole, is endemic to China and is distributed mainly in Qinghai Province. It is of public health interest, as it is, a potential reservoir of and the intermediate host of However, genetic data of this species are lacking, and its name and taxonomy are still a controversy. In the present study, we determined the nucleotide sequence of the entire mitochondrial (mt) genome of and analyzed its evolutionary relationship. The mitogenome was 16328 bp in length and contained 13 protein-coding genes, 22 genes for transfer RNAs (tRNA), two ribosomal RNA genes and two major noncoding regions (O region and D-loop region). Most genes were located on the heavy strand. All tRNA genes had typical cloverleaf structures except for tRNA The mt genome of was rich in A+T (58.45%). Maximum likelihood (ML) and Bayesian methods yielded phylogenetic trees from 33 mt genomes of Arvicolinae, in which formed a sister group with and to the exclusion of species of and other members of the Arvicolinae. Further phylogenetic analyses (ML only) based on the b gene sequences also demonstrated that had a close relationship with The complete mitochondrial genome was successfully assembled and annotated, providing the necessary information for the phylogenetic analyses. Although the name was used in the book 'Wilson & Reeder's Mammal Species of the World', we have confirmed here that its appropriate name is through an analysis of the evolutionary relationships.
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http://dx.doi.org/10.1042/BSR20182349DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6689105PMC
August 2019

Chaihu-Shugan-San and absorbed meranzin hydrate induce anti-atherosclerosis and behavioral improvements in high-fat diet ApoE mice via anti-inflammatory and BDNF-TrkB pathway.

Biomed Pharmacother 2019 Jul 22;115:108893. Epub 2019 Apr 22.

Institute of TCM-Related Comorbid Depression, Nanjing University of Chinese Medicine, Nanjing, 210023, Jiangsu Province, China. Electronic address:

The comorbidity of coronary heart disease (CHD) and depression in patients is extremely prevalent, with a rate from 20 to 50%, while depression-like behaviors exist in a larger percentage of patients. Therefore, the study of comorbidities is particularly urgent. Chaihu-Shugan-San (CSS), a classical traditional Chinese medicine formula, has been used to treat CHD with depression for hundreds of years. However, the mechanism of its action on comorbidities remains unclear. Here, we focused on the behavioral changes in ApoE mice fed a high-fat diet (HFD) and elucidated the mechanism of CSS and its main absorbed component, meranzin hydrate (MH), as an anti-atherosclerosis and anti-depression agent. In the present study, mice were fed an HFD for 16 weeks and were intragastrically administered high and low doses of CSS and MH. Depressive-like behaviors were evaluated by the sucrose preference test, the open-field test, the light-dark test and the tail-suspension test, after which atherosclerotic plaques, plasma lipids, inflammatory cytokine levels and the expression of BDNF/TrkB were measured. We demonstrated that the atherosclerosis model group exhibited significant depressant behaviors. Moreover, CSS inhibited depressive-like behavioral changes, reduced atherosclerotic plaque areas, plasma total cholesterol, triglycerides, LDL-cholesterol levels and inflammatory cytokines including TNF-α, IL-1β, and IL-6 in plasma and hippocampi, increased the protein and mRNA expression of BDNF and TrkB in the aorta and the hippocampus. Interestingly, MH, the main component in CSS that is absorbed in the plasma and hippocampus, exerted effects similar to those of CSS. In addition, MH increased the protein and mRNA expression of BDNF and TrkB in human umbilical vein endothelial cells (HUVECs) induced by LPS. Collectively, these results suggest that MH represents the CSS decoction, induces anti-atherosclerosis effects and improves depression-like behaviors in HFD-fed ApoE mice. Moreover, the regulation of proinflammatory factors and BDNF-TrkB signaling are possibly involved in this process. Our findings indicate that MH is a potential phytochemical compound for the prevention of the comorbidity of atherosclerosis and depression.
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http://dx.doi.org/10.1016/j.biopha.2019.108893DOI Listing
July 2019

Rapid and Visual Detection of Spp. Using a Lateral Flow Strip-Based Recombinase Polymerase Amplification (LF-RPA) Assay.

Front Cell Infect Microbiol 2019 21;9. Epub 2019 Jan 21.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.

spp., are amongst the most widespread parasitic nematodes, primarily live in the muscles of a wide range of vertebrate animals and humans. Human infection occurs by ingestion of raw or undercooked meat containing larvae. Accurate diagnosis of spp. infection in domestic animals is crucial for the effective prevention and control of human trichinellosis. In the present study, a simple, rapid and accurate diagnostic assay was developed combining recombinase polymerase amplification and a lateral flow strip (LF-RPA) to detect spp. infection. The LF-RPA assay targets spp. mitochondrial small-subunit ribosomal RNA () gene and can detect as low as 100 fg DNA of strains, which was approximately 10 times more sensitive than a conventional PCR assay. The LF-RPA assay can be performed within 10-25 min, at a wide range of temperatures (25-45°C) and showed no cross-reactivity with DNA of other parasites and related host species of . The performance of the LF-RPA assay in the presence of high concentration of PCR inhibitor was better than that of a conventional PCR assay. Results obtained by LF-RPA assay for the detection of experimentally infected mice were comparable to the results obtained by using a conventional PCR, achieving 100% specificity and high sensitivity. These results present the developed LF-RPA assay as a new simple, specific, sensitive, rapid and convenient method for the detection of infection in domestic animals.
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http://dx.doi.org/10.3389/fcimb.2019.00001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6348712PMC
December 2019

Genetic Diversity in From the Plateau Vole and Plateau Pika in Jiuzhi County, Qinghai Province, China.

Front Microbiol 2018 5;9:2632. Epub 2018 Nov 5.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Key Laboratory of Veterinary Public Health of Agriculture Ministry, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.

The Qinghai-Tibet Plateau is a highly endemic area of alveolar echinococcosis where a series of intermediate hosts, especially voles and pikas, are infected with . The metacestodes of are fluid-filled, asexually proliferating cysts, and they are mainly found in the host's liver in the form of tumor-like growths. In this study, we investigated the genetic variations of in four mitochondrial (mt) genes, namely, NADH dehydrogenase subunit 5 (5), adenosine triphosphate subunit 6 (6), cytochrome c oxidase subunit 1 (1), and NADH dehydrogenase subunit 1 (1). The complete 5, 6, 1, and 1 genes were amplified separately from each hydatid cyst isolate using polymerase chain reaction (PCR) and then sequenced. Phylogenetic trees were then generated based on the combined mt genes using MrBayes 3.1.2 and PAUP version 4.0b10. The results showed that thirty of 102 voles and two of 49 pikas were infected with . The genetic variation distances among all samples were 0.1-0.4%, 0.2-0.4%, 0.1-0.6%, and 0.1-0.4% for 5, 6, 1, and 1, respectively. Compared to previous studies of the genetic diversity of based on the 1 gene, the genetic distances within the same group were 1.3-1.7% (Mongolia strain), 0.6-0.8% (North American strain), 0.3-0.6% (European strain), and 0.1-0.4% (Asian strain). Based on concatenated sequences of the 5, 6, 1, and 1 genes all haplotypes were divided into two clusters. In conclusion, the genetic diversity of based on mt genes on a small local area is at low level but between different regions with long distance and different ecological environment each other, the genetic diversity is at relatively high level; genetic variation is higher in the 1 gene than that in the other three mt genes. The results on a local scale provide basic information for further study of the molecular epidemiology, genetic differences and control of in Qinghai Province, China.
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http://dx.doi.org/10.3389/fmicb.2018.02632DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6230927PMC
November 2018

Estimating the prevalence of Echinococcus in domestic dogs in highly endemic for echinococcosis.

Infect Dis Poverty 2018 Aug 9;7(1):77. Epub 2018 Aug 9.

The Human Pathology and Immunology Department, Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region, People's Republic of China.

Background: Cystic echinococcosis (CE) and alveolar echinococcosis (AE) are highly endemic in Xiji County of Ningxia Hui Autonomous Region (NHAR) in China where the control campaign based on dog de-worming with praziquantel has been undertaken over preceding decades. This study is to determine the current prevalence of Echinococcus granulosus and E. multilocularis in domestic dogs and monitor the echinococcosis transmission dynamics.

Methods: Study villages were selected using landscape patterns (Geographic Information System, GIS) for Echinococcus transmission "hot spots", combined with hospital records identifying risk areas for AE and CE. A survey of 750 domestic dogs, including copro-sampling and owner questionnaires, from 25 selected villages, was undertaken in 2012. A copro-multiplex PCR assay was used for the specific diagnosis of E. granulosus and E. multilocularis in the dogs. Data analysis, using IBM SPSS Statistics, was undertaken, to compare the prevalence of the two Echinococcus spp. in dogs between four geographical areas of Xiji by the χ test. Univariate analysis of the combinations of outcomes from the questionnaire and copro-PCR assay data was carried out to determine the significant risk factors for dog infection.

Results: The highest de-worming rate of 84.0% was found in the northwest area of Xiji County, and significant differences (P <  0.05) in the de-worming rates among dogs from the four geographical areas of Xiji were detected. The highest prevalence (19.7%, 59/300) of E. multilocularis occurred in northwest Xiji, though the highest prevalence (18.1%, 38/210) of E. granulosus occurred in southwest Xiji. There was no significant difference (P >  0.05) in the prevalence of E. granulosus in dogs from the northwest, southwest, northeast, and southeast of Xiji, but there were significant differences (P <  0.05) between dogs infected with E. multilocularis from the four areas. None of the other independent variables was statistically significant.

Conclusions: The results from this study indicate a high prevalence of both E. granulosus and E. muiltilocularis in dogs in Xiji County, NHAR. Transmission of E. multilocularis was more impacted by geographical risk-factors in Xiji County than that of E. granulosus. Dogs have the potential to maintain the transmission of both species of Echinococcus within local Xiji communities, and the current praziquantel dosing of dogs appears to be ineffective or poorly implemented in this area.
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http://dx.doi.org/10.1186/s40249-018-0458-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6083587PMC
August 2018

Developmental Sensitivity in Schistosoma mansoni to Puromycin To Establish Drug Selection of Transgenic Schistosomes.

Antimicrob Agents Chemother 2018 08 27;62(8). Epub 2018 Jul 27.

Department of Microbiology, Immunology & Tropical Medicine, and Research Center for the Neglected Diseases of Poverty, School of Medicine and Health Sciences, George Washington University, Washington, DC, USA

Schistosomiasis is considered the most important disease caused by helminth parasites, in terms of morbidity and mortality. Tools to facilitate gain- and loss-of-function approaches can be expected to precipitate the discovery of novel interventions, and drug selection of transgenic schistosomes would facilitate the establishment of stable lines of engineered parasites. Sensitivity of developmental stages of schistosomes to the aminonucleoside antibiotic puromycin was investigated. For the schistosomulum and sporocyst stages, viability was quantified by fluorescence microscopy following dual staining with fluorescein diacetate and propidium iodine. By 6 days in culture, the 50% lethal concentration (LC) for schistosomula was 19 μg/ml whereas the sporocysts were 45-fold more resilient. Puromycin potently inhibited the development of -laid eggs (LC, 68 ng/ml) but was less effective against liver eggs (LC, 387 μg/ml). Toxicity for adult stages was evaluated using the xCELLigence-based, real-time motility assay (xWORM), which revealed LCs after 48 h of 4.9 and 17.3 μg/ml for male and female schistosomes, respectively. Also, schistosomula transduced with pseudotyped retrovirus encoding the puromycin resistance marker were partially rescued when cultured in the presence of the antibiotic. Together, these findings will facilitate selection on puromycin of transgenic schistosomes and the enrichment of cultures of transgenic eggs and sporocysts to facilitate the establishment of schistosome transgenic lines. Streamlining schistosome transgenesis with drug selection will open new avenues to understand parasite biology and hopefully lead to new interventions for this neglected tropical disease.
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http://dx.doi.org/10.1128/AAC.02568-17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6105839PMC
August 2018

Genetic diversity in Echinococcus shiquicus from the plateau pika (Ochotona curzoniae) in Darlag County, Qinghai, China.

Infect Genet Evol 2016 11 6;45:408-414. Epub 2016 Jun 6.

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Key Laboratory of Veterinary Public Health of Agriculture Ministry, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease, Yangzhou 225009, Jiangsu Province, PR China. Electronic address:

The metacestode of Echinococcus shiquicus has been recorded previously in the lung and liver of its intermediate host, the plateau pika (Ochotona curzoniae), but there is limited information regarding other organ sites. There is also limited evidence of intra-specific genetic variation within E. shiquicus. A PCR-amplified mitochondrial (mt) nad1 gene fragment (approximately 1400bp in size), with unique EcoRI and SspI restriction sites, was used to distinguish cysts or cyst-like lesions of E. shiquicus from E. multilocularis. Then, the complete mt nad1 and cox1 genes for the E. shiquicus isolates were amplified and sequenced. Phylogenetic tree and haplotype network analyses for the isolates were then generated based on a concatenated dataset of the nad1 and cox1 genes using the neighbour-joining (NJ) method and TCS1.21 software. Nineteen of eighty trapped pikas were found to harbor cysts (71 in total) when dissected at the survey site. Seventeen animals had cysts (fertile) present only in the lungs, one animal had fertile cysts in the lungs and spleen, and one individual had an infertile kidney cyst. Restriction endonuclease analysis of a fragment of the nad1 gene indicated all the cysts were due to E. shiquicus. Genetic diversity analysis revealed that the nad1 and cox1 genes varied by 0.1-1.2% and 0.1-1.0%, respectively. Haplotype network analysis of the concatenated nad1 and cox1 sequences of the isolates showed they were classified into at least 6 haplotypes, and different haplotype percentages ranged from 4.2% to 29.6%. Although, high haplotype diversity was evident in the study area, the complete nad1 and cox1 gene sequences obtained indicated that all samples represented isolates of E. shiquicus. The study has also provided a new PCR-restriction endonuclease-based method to rapidly distinguish E. shiquicus from E. multilocularis which provides a useful tool for epidemiological investigations where the two species overlap.
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http://dx.doi.org/10.1016/j.meegid.2016.06.016DOI Listing
November 2016

HIV-1 Integrates Widely throughout the Genome of the Human Blood Fluke Schistosoma mansoni.

PLoS Pathog 2016 Oct 20;12(10):e1005931. Epub 2016 Oct 20.

Department of Microbiology, Immunology & Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine and Health Sciences, The George Washington University, Washington, DC, United States of America.

Schistosomiasis is the most important helminthic disease of humanity in terms of morbidity and mortality. Facile manipulation of schistosomes using lentiviruses would enable advances in functional genomics in these and related neglected tropical diseases pathogens including tapeworms, and including their non-dividing cells. Such approaches have hitherto been unavailable. Blood stream forms of the human blood fluke, Schistosoma mansoni, the causative agent of the hepatointestinal schistosomiasis, were infected with the human HIV-1 isolate NL4-3 pseudotyped with vesicular stomatitis virus glycoprotein. The appearance of strong stop and positive strand cDNAs indicated that virions fused to schistosome cells, the nucleocapsid internalized and the RNA genome reverse transcribed. Anchored PCR analysis, sequencing HIV-1-specific anchored Illumina libraries and Whole Genome Sequencing (WGS) of schistosomes confirmed chromosomal integration; >8,000 integrations were mapped, distributed throughout the eight pairs of chromosomes including the sex chromosomes. The rate of integrations in the genome exceeded five per 1,000 kb and HIV-1 integrated into protein-encoding loci and elsewhere with integration bias dissimilar to that of human T cells. We estimated ~ 2,100 integrations per schistosomulum based on WGS, i.e. about two or three events per cell, comparable to integration rates in human cells. Accomplishment in schistosomes of post-entry processes essential for HIV-1replication, including integrase-catalyzed integration, was remarkable given the phylogenetic distance between schistosomes and primates, the natural hosts of the genus Lentivirus. These enigmatic findings revealed that HIV-1 was active within cells of S. mansoni, and provided the first demonstration that HIV-1 can integrate into the genome of an invertebrate.
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http://dx.doi.org/10.1371/journal.ppat.1005931DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5072744PMC
October 2016

Discrimination between E. granulosus sensu stricto, E. multilocularis and E. shiquicus Using a Multiplex PCR Assay.

PLoS Negl Trop Dis 2015 Sep 22;9(9):e0004084. Epub 2015 Sep 22.

State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veterinary Parasitology of Gansu Province/Key Laboratory of Zoonoses of Agriculture Ministry/Lanzhou Veterinary Research Institute, CAAS, Lanzhou, People's Republic of China.

Background: Infections of Echinococcus granulosus sensu stricto (s.s), E. multilocularis and E. shiquicus are commonly found co-endemic on the Qinghai-Tibet plateau, China, and an efficient tool is needed to facilitate the detection of infected hosts and for species identification.

Methodology/principal Findings: A single-tube multiplex PCR assay was established to differentiate the Echinococcus species responsible for infections in intermediate and definitive hosts. Primers specific for E. granulosus, E. multilocularis and E. shiquicus were designed based on sequences of the mitochondrial NADH dehydrogenase subunit 1 (nad1), NADH dehydrogenase subunit 5 (nad5) and cytochrome c oxidase subunit 1 (cox1) genes, respectively. This multiplex PCR accurately detected Echinococcus DNA without generating nonspecific reaction products. PCR products were of the expected sizes of 219 (nad1), 584 (nad5) and 471 (cox1) bp. Furthermore, the multiplex PCR enabled diagnosis of multiple infections using DNA of protoscoleces and copro-DNA extracted from fecal samples of canine hosts. Specificity of the multiplex PCR was 100% when evaluated using DNA isolated from other cestodes. Sensitivity thresholds were determined for DNA from protoscoleces and from worm eggs, and were calculated as 20 pg of DNA for E. granulosus and E. shiquicus, 10 pg of DNA for E. multilocularis, 2 eggs for E. granulosus, and 1 egg for E. multilocularis. Positive results with copro-DNA could be obtained at day 17 and day 26 after experimental infection of dogs with larval E. multilocularis and E. granulosus, respectively.

Conclusions/significance: The multiplex PCR developed in this study is an efficient tool for discriminating E. granulosus, E. multilocularis and E. shiquicus from each other and from other taeniid cestodes. It can be used for the detection of canids infected with E. granulosus s.s. and E. multilocularis using feces collected from these definitive hosts. It can also be used for the identification of the Echinococcus metacestode larva in intermediate hosts, a stage that often cannot be identified to species on visual inspection.
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http://dx.doi.org/10.1371/journal.pntd.0004084DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4578771PMC
September 2015