title: Seroprevalence, isolation and co-infection of multiple Toxoplasma gondii strains in individual bobcats (Lynx rufus) from Mississippi, USA.
abstract: Toxoplasma gondii causes lifelong chronic infection in both feline definitive hosts and intermediate hosts. Multiple exposures to the parasite are likely to occur in nature due to high environmental contamination. Here, we present data of high seroprevalence and multiple T. gondii strain co-infections in individual bobcats (Lynx rufus). Unfrozen samples (blood, heart, tongue and faeces) were collected from 35 free ranging wild bobcats from Mississippi, USA. Toxoplasma gondii antibodies were detected in serum by the modified agglutination test (1:≥200) in all 35 bobcats. Hearts from all bobcats were bioassayed in mice and viable T. gondii was isolated from 21; these strains were further propagated in cell culture. Additionally, DNA was extracted from digests of tongues and hearts of all 35 bobcats; T. gondii DNA was detected in tissues of all 35 bobcats. Genetic characterisation of DNA from cell culture-derived isolates was performed by multiplex PCR using 10 PCR-RFLP markers. Results showed that ToxoDB genotype #5 predominated (in 18 isolates) with a few other types (#24 in two isolates, and #2 in one isolate). PCR-DNA sequencing at two polymorphic markers, GRA6 and GRA7, detected multiple recombinant strains co-infecting the tissues of bobcats; most possessing Type II alleles at GRA7 versus Type X (HG-12) alleles at GRA6. Our results suggest that individual bobcats have been exposed to more than one parasite strain during their life time.
authors.affiliation: United States Department of Agriculture, Agricultural Research Service, Beltsville Agricultural Research Center, Animal Parasitic Diseases Laboratory, Building 1001, Beltsville, MD 20705-2350, USA.
authors.forename: Shiv K
authors.initials: SK
authors.lastname: Verma
authors.affiliation: Molecular Parasitology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Amy R
authors.initials: AR
authors.lastname: Sweeny
authors.affiliation: Game Mammals Section, Bureau of Wildlife Management, Pennsylvania Game Commission, 2001 Elmerton Ave, Harrisburg, PA 17110, USA.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Lovallo
authors.affiliation: United States Department of Agriculture, Agricultural Research Service, Beltsville Agricultural Research Center, Animal Parasitic Diseases Laboratory, Building 1001, Beltsville, MD 20705-2350, USA.
authors.forename: Rafael
authors.initials: R
authors.lastname: Calero-Bernal
authors.affiliation: United States Department of Agriculture, Agricultural Research Service, Beltsville Agricultural Research Center, Animal Parasitic Diseases Laboratory, Building 1001, Beltsville, MD 20705-2350, USA.
authors.forename: Oliver C
authors.initials: OC
authors.lastname: Kwok
authors.affiliation: Department of Microbiology, University of Tennessee, Knoxville, TN 37996-0845, USA.
authors.forename: Tiantian
authors.initials: T
authors.lastname: Jiang
authors.affiliation: Department of Microbiology, University of Tennessee, Knoxville, TN 37996-0845, USA.
authors.forename: Chunlei
authors.initials: C
authors.lastname: Su
authors.affiliation: Molecular Parasitology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA. Electronic address: griggm@niaid.nih.gov.
authors.forename: Michael E
authors.initials: ME
authors.lastname: Grigg
authors.affiliation: United States Department of Agriculture, Agricultural Research Service, Beltsville Agricultural Research Center, Animal Parasitic Diseases Laboratory, Building 1001, Beltsville, MD 20705-2350, USA. Electronic address: Jitender.Dubey@ars.usda.gov.
authors.forename: Jitender P
authors.initials: JP
authors.lastname: Dubey
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000953:Antigens, Protozoan / Q000235:genetics; D001483:Base Sequence; D002415:Cats; D002522:Chlorocebus aethiops; D060085:Coinfection / Q000469:parasitology / Q000662:veterinary*; D016054:DNA, Protozoan / Q000235:genetics; D005260:Female; D005838:Genotype; D046008:Lynx / Q000469:parasitology*; D008297:Male; D051379:Mice; D008922:Mississippi / Q000453:epidemiology; D011110:Polymorphism, Genetic; D015800:Protozoan Proteins / Q000235:genetics; D016036:Seroepidemiologic Studies; D014122:Toxoplasma / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D014124:Toxoplasmosis, Animal / Q000175:diagnosis / Q000453:epidemiology* / Q000469:parasitology*
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D016054:DNA, Protozoan; C095047:GRA6 protein, Toxoplasma gondii; C112573:GRA7 protein, Toxoplasma gondii; D015800:Protozoan Proteins
keywords: Bioassay; Felids; Genotype; Isolation; Strain; Toxoplasmosis