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      Diagnosis of Swine Toxoplasmosis by PCR and Genotyping of Toxoplasma gondii from pigs in Henan, Central China

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          Abstract

          Background

          Toxoplasma gondii, a widely prevalent protozoan parasite, causes serious toxoplasmosis infections in humans and other animals. Among livestock, pigs are susceptible to T. gondii infection. Despite Henan being one of the biggest pig-raising provinces in China, little information exists on the epidemiology of toxoplasmosis in this location. Therefore, we molecularly characterized DNA samples from pigs in Henan. A total of 1647 samples, including 952 from dead piglets, 478 from seriously sick fattening pigs and 217 from abortion sows, were collected from different animal hospitals or pig farms from 10 different cities in Henan (2006–2008). Each pig corresponded to a separate pig farm. DNA was extracted from 3 to 5 g of the most severely affected pig tissue (liver, spleen, lung, hilar lymph nodes and amniotic fluid) after postmortem examination. The presence of the T. gondii B1 gene was detected using nested polymerase chain reactions (PCR). Genotyping was performed directly on DNA from the PCR-positive tissue samples using 11 PCR restriction fragment length polymorphism markers (SAG1, 5′- and 3′-SAG2, alternative SAG2, SAG3, BTUB, GRA6, L358, PK1, c22–8, c29–2, and Apico).

          Results

          Of all samples, thirty-four were positive for the T. gondii B1 gene (2.06%, 95% CI: 1.86%–2.26%) from four cities, including 31 from NanYang city, one (PgXY 1) from Xinyang City, one (PgZZ 1) from Zhengzhou City and one (PgZK1) from Zhoukou City. The prevalence was found to be highest in piglets than in fattening pigs and sows. And the difference was statistically significant (P<0.01). The following 32 samples were genotyped with complete data: 13 hilar lymph node tissue samples, seven liver tissue samples, seven lung tissue samples, four spleen tissue samples, and one amniotic fluid sample. Only one genotype, belonging to ToxoDB Genotype #9, was identified.

          Conclusions

          This is the first large-scale survey molecularly characterizing T. gondii from pigs in Henan. The results of the present study revealed that T. gondii infection is present in swine in Henan and is a potential source of foodborne toxoplasmosis in the investigated areas. Implementation of effective control measures for T. gondii to reduce the chance of zoonotic toxoplasmosis spreading from pig farms may be warranted. The results show that the ToxoDB #9 genotype may be the dominant T. gondii lineage in mainland China. These findings strengthen the limited Chinese T. gondii epidemiology database.

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          Most cited references35

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          Toxoplasmosis: A history of clinical observations.

          It has been 100 years since Toxoplasma gondii was initially described in Tunis by Nicolle and Manceaux (1908) in the tissues of the gundi (Ctenodoactylus gundi) and in Brazil by Splendore (1908) in the tissues of a rabbit. Toxoplasma gondii is a ubiquitous, Apicomplexan parasite of warm-blooded animals that can cause several clinical syndromes including encephalitis, chorioretinitis, congenital infection and neonatal mortality. Fifteen years after the description of T. gondii by Nicolle and Manceaux a fatal case of toxoplasmosis in a child was reported by Janků. In 1939 Wolf, Cowen and Paige were the first to conclusively identify T. gondii as a cause of human disease. This review examines the clinical manifestations of infection with T. gondii and the history of the discovery of these manifestations.
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            Toxoplasma gondii comprises three clonal lineages: correlation of parasite genotype with human disease.

            The population genetic structure of Toxoplasma gondii was determined by multilocus restriction fragment length polymorphism analysis at 6 loci in 106 independent isolates from humans and animals. Phylogenetic and statistical analyses indicated a highly unusual population structure consisting of 3 widespread clonal lineages. Extensively mixed genotypes were only apparent in 4 strains, which indicated that, while not separate species, sexual recombination between the 3 lineages is exceedingly rare in natural populations. T. gondii is a major cause of subclinical human infection and an important opportunistic pathogen that causes severe disease in immunocompromised patients. While strains from all 3 lineages were isolated from humans, the majority of human toxoplasmosis cases were associated with strains of a type II genotype. The correlation of specific clonal lineages with human toxoplasmosis has important implications for development of vaccines, drug treatments, and diagnostic protocols.
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              Moving towards an integrated approach to molecular detection and identification of Toxoplasma gondii.

              The development of simple, sensitive and rapid methods for the detection and identification of Toxoplasma gondii is important for the diagnosis and epidemiological studies of the zoonotic disease toxoplasmosis. In the past 2 decades, molecular methods based on a variety of genetic markers have been developed, each with its advantages and limitations. The application of these methods has generated invaluable information to enhance our understanding of the epidemiology, population genetics and phylogeny of T. gondii. However, since most studies focused solely on the detection but not genetic characterization of T. gondii, the information obtained was limited. In this review, we discuss some widely used molecular methods and propose an integrated approach for the detection and identification of T. gondii, in order to generate maximum information for epidemiological, population and phylogenetic studies of this key pathogen.
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                Author and article information

                Contributors
                wangheng800220@126.com
                zhanglx8999@gmail.com , zhanglongxian8999@foxmail.com
                1115110106@qq.com
                yufuchang456@qq.com
                yyrhenau@foxmail.com
                Journal
                BMC Vet Res
                BMC Vet. Res
                BMC Veterinary Research
                BioMed Central (London )
                1746-6148
                31 May 2017
                31 May 2017
                2017
                : 13
                : 152
                Affiliations
                [1 ]GRID grid.108266.b, College of Animal Science and Veterinary Medicine, , Henan Agricultural University, ; Zhengzhou, 450002 Henan People’s Republic of China
                [2 ]Department of Animal Science, Henan Vocational College of Agriculture, Zhongmu, 451450 Henan, People’s Republic of China
                Author information
                http://orcid.org/0000-0002-5706-131X
                Article
                1079
                10.1186/s12917-017-1079-3
                5452427
                2cbac630-f21c-46ab-8289-23df19a54f9e
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 3 November 2016
                : 26 May 2017
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 31330079
                Award Recipient :
                Funded by: Innovation Scientists and Technicians Troop Construction Projects of Henan Province
                Award ID: 134200510012
                Award Recipient :
                Funded by: Key National Science and Technology Specific Projects
                Award ID: 2012ZX10004220-011
                Award Recipient :
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2017

                Veterinary medicine
                swine,toxoplasmosis,pcr assay,diagnosis,molecular characterization,henan
                Veterinary medicine
                swine, toxoplasmosis, pcr assay, diagnosis, molecular characterization, henan

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