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Molecular detection of Toxoplasma gondii in chicken hearts from markets and retail stores in Northern Iran.

ABSTRACT: Detection of Toxoplasma gondii in chicken products indicates risk of transmission to consumers. The objective of the current study was to investigate the molecular prevalence of T. gondii in free-ranging and industrial chickens in Guilan province, Northern Iran. A total of 150 chicken heart samples including 75 free-range and 75 industrial chickens were collected from farmers' markets and chicken retailers in Guilan, Northern Iran, between October 2017 and August 2018. Genomic DNA were extracted from samples and examined for evidence of T. gondii using polymerase chain reaction (PCR) targeting the B1 gene. The B1-positive samples were further analyzed by nested-PCR for SAG1 gene. Of the 150 samples, T. gondii DNA fragments were detected in 59 (39.3%), including 30 (40%) free-range and 29 (38.7%) industrial chicken. No significant differences of T. gondii DNA detection was observed between the free-range and industrial chicken samples (p = 0.73). Four selected positive samples were used for amplifying and sequencing of the SAG1 gene. The results revealed that all four sequences of SAG1 had 100% similarity with T. gondii sequences previously isolated from an AIDS/HIV patient in Mazandaran province, Northern Iran. Furthermore, the phylogenetic analysis demonstrated that all four sequences were closely related to Type I of T. gondii. However, our Type I identification is preliminary and needs to be confirmed by further multilocus sequence typing (MLST) analysis. The findings of the present study provide new data about the presence of T. gondii DNA in chicken hearts in the study area. These results confirm that chicken can be used as sentinels for environment contamination; however, further studies are needed to determine the viability of T. gondii in chicken hearts from Iran for risk assessment.

SUBMITTER: Abbaszadeh S

PROVIDER: S-EPMC9249558 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Avibacterium (Haemophilus) Paragallinarum (Av. Paragallinarum) is the causative agent of Infectious Coryza (IC) in chickens. Despite the worldwide distribution of IC, no systematic study, to the best of our knowledge, was conducted on isolation and characterization of Av. Paragallinarum in Iran. The present study aimed to isolate and perform antibacterial susceptibility testing (AST) of IC agents from suspected backyard chickens with typical symptoms of IC in avian markets. From 18 collected choanal swab samples, four (22%) isolates of Av. Paragallinarum were detected by culture methods based on satellite growth on blood agar, which was confirmed by the biochemical reaction of Catalase and Oxidase tests and species-specific PCR (HPG-2). The hypervariable region of the hemagglutinin genes of 4 isolates was amplified and obtained sequences were deposited at a gene bank for more characterization. Meanwhile, 12 (66%) positive reactions were detected by observing expected 500 bpb and using PCR (HPG-2) on swab samples. Antibiotic susceptibility testing (AST) of obtained isolates were analyzed using the Kirby-Bauer disk diffusion method on Columbia agar with horse blood. Isolates were found to be resistant to amoxicillin, oxytetracycline, streptomycin, trimethoprim/sulfamethoxazole (up to 75%) and sensitive to cefalexin, ceftriaxone, enrofloxacin, florfenicol, gentamycin, linco-spectin, neomycin, doxycycline (50%), danofloxacin (75%), flumequine (50%), ofloxacin (75%). An intermediate growth inhibitionzone has been observed around antibiotic discs for ampicillin, colistin, erythromycin, penicillin, tiamulin (75%), tylosin (75%). In summary, to the best of our knowledge, this study is the first report of isolation and identification of Avibacterium Paragallinarum from backyard chickens which may be a source of IC for commercial chicken flocks. Moreover, the prevalence of resistance to some antibacterial drugs of IC agents may impose an additional threat to the poultry industry. A more in-depth study is recommended to develop a low-cost autogenous IC vaccine for small-scale flocks of poultry to prevent and manage the disease and establish antimicrobial resistance.

Project description:BackgroundThe consumption of uncooked or undercooked food from infected intermediate hosts can result in Toxoplasma gondii infection in humans. However, few studies have investigated the genetic diversity of this protozoan parasite in Iran. The aim of the present study was to genetically characterize isolates of T. gondii from intermediate host animals in Mazandaran Province, Iran.MethodsBlood and heart tissue samples were collected from 204 ruminants, and brain tissue was collected from 335 birds. The prevalence of T. gondii infection in these samples was determined serologically using the modified agglutination test and by conventional PCR assays. Those PCR samples positive for T. gondii DNA and 13 DNA samples extracted from aborted fetuses in a previous study were genotyped with 12 genetic markers using the multilocus-nested PCR-restriction fragment length polymorphism (Mn-PCR-RFLP) technique.ResultsAntibodies for parasites were found in 35.7% of the ruminant (39.1% of sheep and 26.4% of goats) samples and in 51.3% of the bird (100% of geese, 52.3% of free-range chickens and 46% of ducks) samples. Molecular detection by PCR of the repetitive 529-bp DNA fragment revealed contamination of 13.2% of ruminant (14.6% of sheep and 9.4% of goats) samples and of 9.6% of bird (11.1% of chickens, 5.7% of ducks and 0% of geese samples). The results from 30 DNA samples revealed five ToxoDB (genome database for the genus Toxoplasma) PCR-RFLP genotypes, including #1 (Type II), #2 (Type III), #10 (Type I), #27 and #48, with genotype #1 the most predominant.ConclusionsAs evidenced by the results of this study, ruminants and birds are infected with T. gondii in the region, suggesting that they are likely to be involved in the transmission of T. gondii to humans through meat consumption. The identification of different genotypes may suggest a higher genetic diversity of this parasite in Mazandaran, reflecting local environmental contamination. These results have important public health implications for the prevention and control strategies of infection.

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Molecular detection of Toxoplasma gondii in chicken hearts from markets and retail stores in Northern Iran.

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