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Prevalence of Toxoplasma gondii infection in rabbits of Korea by serological tests and nested polymerase chain reaction.

ABSTRACT: This study surveyed the Toxoplasma (T.) gondii infection prevalence in the Korean rabbit population. Rabbits (n=142) were obtained from two breeding farms in the Gongju area, Chungnam Province, and in the Kochang area, Junbuk Province, Korea. Of 142 sera samples analyzed by enzyme-linked immunosorbent assay (ELISA), 15 (10.6%) exhibited T. gondii-specific IgG antibodies, and 1 (0.7%) rabbit harbored T. gondii-specific IgM. Female rabbits (9/84; 10.7%) had a similar T. gondii prevalence to males (6/58; 10.3%). When stratified by age, rabbits aged >1 year had a similar prevalence of T. gondii infection (7/66; 10.6%) to rabbits aged <1 year (8/76; 10.5%). Immunoblotting detected 6 major antigenic bands corresponding to T. gondii-positive sera at 20, 28, 30, 35, 63 and 77 kDa. Nested polymerase chain reaction (PCR) of whole-blood samples detected the T. gondii B1 gene in 23 rabbits (16.2%). All PCR-positive samples corresponded to partial T. gondii B1 gene sequences with 99% homology to a T. gondii sequence deposited in GenBank (accession number EU340874). Female rabbits (13/84; 15.5%) harbored a similar prevalence of T. gondii DNA to males (10/58; 17.2%). Rabbits aged >1 year had a similar prevalence (12/66; 18.2%) of T. gondii infection to rabbits aged <1 year (11/76; 14.5%). No statistically significant differences were observed regarding the prevalences of infection according to sex or age using molecular or serological tests. This study is the first survey using serological tests and nested PCR to analyze the T. gondii prevalence in rabbits in Korea.

SUBMITTER: Shin HG

PROVIDER: S-EPMC3942966 | biostudies-literature | 2013 Dec

REPOSITORIES: biostudies-literature

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Prevalence of Toxoplasma gondii infection in rabbits of Korea by serological tests and nested polymerase chain reaction.

Shin Hyun-Guk HG Lee Sang-Eun SE Hong Sung-Hee SH Kim Se-Mi SM Choi Young-Ki YK Park Hyung-Jin HJ Seo Kyoung-Won KW Song Kun-Ho KH

The Journal of veterinary medical science 20130813 12

This study surveyed the Toxoplasma (T.) gondii infection prevalence in the Korean rabbit population. Rabbits (n=142) were obtained from two breeding farms in the Gongju area, Chungnam Province, and in the Kochang area, Junbuk Province, Korea. Of 142 sera samples analyzed by enzyme-linked immunosorbent assay (ELISA), 15 (10.6%) exhibited T. gondii-specific IgG antibodies, and 1 (0.7%) rabbit harbored T. gondii-specific IgM. Female rabbits (9/84; 10.7%) had a similar T. gondii prevalence to males ...[more]

PMID: 23955398

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Project description:IntroductionHammondia hammondi and Toxoplasma gondii are closely related protozoan parasites, but only T. gondii is zoonotic. Both species use felids as definitive hosts and cannot be differentiated by oocyst morphology. In T. gondii, a 529-base pair (bp) repetitive element (TgREP-529) is of utmost diagnostic importance for polymerase chain reaction (PCR) diagnostic tests. We identified a similar repetitive region in the H. hammondi genome (HhamREP-529).MethodsBased on reported sequences, primers and probes were selected in silico and optimal primer probe combinations were explored, also by including previously published primers. The analytical sensitivity was tested using serial dilutions of oocyst DNA. For testing analytical specificity, DNA isolated from several related species was used as controls. The newly established TaqMan PCR (Hham-qPCR1) was applied to tissues collected from H. hammondi-infected gamma-interferon gene knockout (GKO) mice at varying time points post-infection.ResultsTen forward and six reverse primers were tested in varying combinations. Four potentially suitable dual-labelled probes were selected. One set based on the primer pair (Hham275F, Hham81R) and the probe (Hham222P) yielded optimal results. In addition to excellent analytic specificity, the assay revealed an analytical sensitivity of genome equivalents of less than one oocyst. Investigation of the tissue distribution in GKO mice revealed the presence of parasite DNA in all examined organs, but to a varying extent, suggesting 100- to 10,000-fold differences in parasitic loads between tissues in the chronic state of infection, 42 days post-infection.DiscussionThe use of the 529-bp repeat of H. hammondi is suitable for establishing a quantitative real-time PCR assay, because this repeat probably exists about 200 times in the genome of a single organism, like its counterpart in T. gondii. Although there were enough sequence data available, only a few of the primers predicted in silico revealed sufficient amplification; the identification of a suitable probe was also difficult. This is in accord with our previous observations on considerable variability in the 529-bp repetitive element of H. hammondi.ConclusionsThe H. hammondi real-time PCR represents an important novel diagnostic tool for epidemiological and cell biological studies on H. hammondi and related parasites.

Project description:BackgroundLittle is known about the epidemiology of Toxoplasma gondii and Neospora caninum infections in alpacas (Vicugna pacos) and llamas (Lama glama) outside South America. The study aimed to estimate the seroprevalence of T. gondii and N. caninum infections in South American camelids (SAC) in Switzerland, to optimize serological tests for SAC and to identify risk factors, which may favour infection.MethodsA total of 571 sera from 132 Swiss farms (374 alpacas and 197 llamas, mean 4.3 animals/farm) were obtained. Four commercial enzyme-linked immunosorbent assays (ELISA) for detecting antibodies against T. gondii (ID Screen® Toxoplasmosis Indirect (TOXO-MS)) or N. caninum (i.e. ID Screen® Neospora caninum Indirect Multi-species (NCS-MS); ID Screen® Neospora caninum Competition (NCC) and ID Screen® Neospora caninum Indirect (NCS)) were first assessed for their use on SAC comparing their results with those in immunoblot, and optimizing cut-offs. Subsequently, two kits (TOXO-MS and NCS-MS) were selected for seroprevalence estimation. Additionally, a risk factor analysis for infection was performed on 41 farms, which agreed to participate in a web-based survey.ResultsThree kits (TOXO-MS, NCS-MS and NCC) showed almost perfect agreement (kappa > 0.901) with immunoblot results when the cut-offs were optimized, and one kit (NCS) proved not to be useful for detecting N. caninum seropositive SAC. By TOXO-MS ELISA, 82.3% (308/374) of the alpacas and 84.8% (167/197) of the llamas were seropositive for T. gondii, and 131/132 (99.2%) farms had seropositive animals. By NCS-MS ELISA, 3.5% (13/374) of the alpacas and 2.5% (5/197) of the llamas evidenced antibodies against N. caninum, and 9.1% (12/132) of the farms had seropositive animals. The variables "age" and "female sex" were identified as risk factors for T. gondii infection and "absence of cats in the farm during the last two years" as a protective factor. No risk or protective factors for N. caninum infection could be identified.ConclusionsThis nationwide cross-sectional study demonstrated for the first time the presence of antibodies against T. gondii and N. caninum in the Swiss SAC population, highlighting a high seroprevalence for T. gondii, the presence of cats as a risk factor and suggesting that SAC meat might represent an additional infection source for humans.

Dataset Information

Prevalence of Toxoplasma gondii infection in rabbits of Korea by serological tests and nested polymerase chain reaction.

Publications

Prevalence of Toxoplasma gondii infection in rabbits of Korea by serological tests and nested polymerase chain reaction.

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