Open Veterinary Journal, (2026), Vol. 16(4): 2062-2071

Research Article

10.5455/OVJ.2026.v16.i4.11

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Prevalence of Toxoplasma gondii in Chinese stray dogs: a meta-analysis

Chaoyang Chen^1,2* and Xiaoxia Ji³

¹Department of Medical Science and Technology, Suzhou Chien-Shiung Institute of Technology, Suzhou, China

²Jiangsu Province Engineering Research Center of Novel Tumor-targeting Drug Conjugates, Suzhou, China

³Crown Bioscience Inc., Suzhou, China

*Corresponding Author: Chaoyang Chen. Department of Medical Science and Technology, Suzhou Chien-Shiung Institute of Technology, Suzhou, China. Email: 271854008 [at] qq.com

Submitted: 10/11/2025 Revised: 28/02/2026 Accepted: 14/03/2026 Published: 30/04/2026

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Abstract

Background: Toxoplasma gondii is a globally distributed zoonotic parasite infecting nearly all warm-blooded animals. However, data on the prevalence of T. gondii in stray dogs across China remain fragmented.

Aim: To estimate the pooled prevalence of T. gondii in stray dogs in China.

Methods: Five databases (PubMed, China National Knowledge Infrastructure, Wanfang, VIP, and Baidu Scholar) were searched for studies reporting the serological or molecular detection of T. gondii in stray dogs. A random-effects model was used to calculate pooled prevalence. Subgroup analyses were performed according to sex, age, detection method, period, and region. Sensitivity analysis and publication bias were performed to assess study robustness.

Results: Seventeen studies (2009–2022) involving 2,320 stray dogs from 14 provinces were included. The pooled seroprevalence of T. gondii was 31% (95% confidence interval: 22%–40%). No significant differences were found among sex, age, region, or study period (p > 0.05), whereas seroprevalence estimates were significantly higher in studies using ELISA compared with IHA (Q=19.24, df=1, p < 0.0001). Only three studies detected T. gondii DNA, with reported positivity rates ranging from 2% to 47%, precluding pooled estimation.

Conclusion: Toxoplasma gondii infection is widespread among stray dogs in China, highlighting the need for strengthened surveillance and integrated control measures.

Keywords: China, Molecular detection, Seroprevalence, Stray dogs, Toxoplasma gondii.

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Introduction

Toxoplasma gondii (T. gondii) is an obligate intracellular protozoan parasite with a global distribution and the ability to infect nearly all warm-blooded animals, including humans. The parasite’s definitive hosts are felids, which excrete environmentally resistant oocysts in their feces. Intermediate hosts, such as dogs and livestock, become infected through ingestion of oocysts or consumption of raw or undercooked meat containing tissue cysts. In humans, T. gondii infection can cause life-threatening complications in immunocompromised individuals as well as miscarriage, stillbirth, premature birth, fetal malformations in pregnant women (Bobić et al., 2019).

Dogs play a multifaceted role in the epidemiology of T. gondii. Although dogs are not definitive hosts of T. gondii, they may act as intermediate hosts or mechanical carriers, facilitating indirect environmental dissemination and increasing the risk of exposure to humans and other animals (Lindsay et al., 1997). Since the first report of canine toxoplasmosis in Italy in 1910 (Jacobs et al., 1955), the parasite in dogs has been identified globally, with relatively high infection rates (Hamidullah et al., 2022). Stray dogs, in particular, often scavenge and roam freely, encountering contaminated environments and potential hosts, thereby exhibiting higher infection rates than owned or confined dogs (Otranto et al., 2017). In some areas of the world, dog meat is consumed by humans or fed to felids, potentially facilitating T. gondii transmission if undercooked meat is ingested (Dubey et al., 2020).

In China, T. gondii infection is widespread among various animal species, including pigs, sheep, poultry, and dogs. Human seroprevalence remains relatively high in some regions of China (Zhou et al., 2011; Mao et al., 2021; Su et al., 2022; Yang et al., 2024; Wang et al., 2025). Several studies have investigated T. gondii infection in dogs, but most have focused on pet or owned dogs (Gao et al., 2016; Cong et al., 2018; Sheng et al., 2020; Wen et al., 2025). Stray dogs, which often live in proximity to human communities but lack veterinary care or vaccination, may better reflect the environmental burden of T. gondii oocysts. However, no comprehensive synthesis has been conducted to evaluate the infection status of stray dogs across China.This study aimed to systematically review and quantitatively analyze the prevalence of T. gondii infection in stray dogs in China.

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Materials and Methods

Search strategy

This systematic review and meta-analysis was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines (Page et al., 2021). Notably, a protocol was not pre-registered due to the exploratory nature of this review at inception; however, we strictly adhered to Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines post hoc to ensure transparency, though this may limit prospective reproducibility. A comprehensive search strategy was employed to identify relevant studies. For Chinese literature, we queried the China National Knowledge Infrastructure, Wanfang Data, and CQVIP databases. In the English-language realm, we leveraged PubMed and Baidu Scholar. To capture gray literature, we included dissertations, purposefully seeking out unpublished data to enrich our analysis. The search terms were constructed based on three key elements: study subjects, pathogens, and geographical location. For study subjects, terms such as "stray dogs", "street dogs", "free-roaming dogs", "feral dogs" and their Chinese equivalents were used. Pathogen-specific terms included “Toxoplasma gondii” OR “toxoplasmosis”. Geographical terms included "China", "Chinese". The search strings combined these elements using Boolean operators. No language or publication date restrictions were applied during the initial search. Search results were imported into Zotero: those from English-language sources, such as PubMed, were directly integrated using the native PubMed format, while citations from Chinese databases were converted via RefWorks before being added to Zotero. The search ended on June 15, 2025, with all databases searched on the same day.

Inclusion and exclusion criteria

Studies were included if they met all of the following criteria:

(1) The target population consisted of stray, free-roaming, feral, shelter, or street dogs in China; (2) Toxoplasma gondii infection was confirmed by serological methods or molecular assays; (3) The sample size and number of positive cases were reported or could be derived; and (4) The study provided traceable information on the time and location of the investigation.

The exclusion criteria were as follows:

(1) Studies limited to pet or owned dogs; (2) Case reports, reviews, or conference abstracts without primary data; (3) Studies lacking clear diagnostic methods or sample details; and (4) Duplicate reports from the same population or dataset, in which case the most complete or recent version was retained.

After importing the search results into Zotero, all duplicate entries were removed. Reviewers CC and XJ independently screened the titles and abstracts of all retrieved records according to predefined inclusion and exclusion criteria. Prior to formal screening, the reviewers discussed the eligibility criteria in detail and screened a small subset of records to ensure a shared understanding of the selection standards. Given the clearly defined criteria and the relatively homogeneous study designs, no formal inter-reviewer agreement statistics were calculated. Full-text articles were subsequently assessed independently by both reviewers. Discrepancies at any stage were resolved through discussion, and consensus was reached in all cases.

Data extraction

Two reviewers independently extracted relevant information using a standardized data collection form. Extracted variables included the following: first author, publication year, study period, study location (province or city), sample type, sample size, number of positives, detection method, and diagnostic target (antibody, antigen, or DNA). The two datasets were compared after extraction, and any discrepancies were resolved through discussion to ensure accuracy and consistency.

Quality assessment

Study quality was assessed using a modified version of the Joanna Briggs Institute critical appraisal checklist (Munn et al., 2017). Key criteria were evaluated, including sampling representativeness, diagnostic validity, reporting completeness, and statistical appropriateness. Each domain was scored on a 0–2 scale, yielding a total quality score ranging from 0 to 8. Studies scoring ≥ 6 were considered high quality, typically characterized by clearly defined sampling strategies, validated diagnostic methods, and adequate reporting of key methodological details. Studies scoring ≤ 3 were considered low quality, typically due to unclear sampling procedures, insufficient methodological description, or use of unvalidated assays.

Data analysis

Meta-analyses were performed using the R software (version 4.3.2) with the “meta” and “metafor” packages (Balduzzi et al., 2019). To stabilize variance, prevalence data were transformed using the Freeman–Tukey double arcsine method. The pooled prevalence and 95% confidence interval (CI) were estimated using a random-effects model (DerSimonian–Laird method), accounting for between-study heterogeneity. Heterogeneity was quantified using the Cochran’s Q test (chi-squared) and the Higgins I² statistic, with I² values of 25%, 50%, and 75% representing low, moderate, and high heterogeneity, respectively. When applicable, subgroup analyses were conducted based on diagnostic method, geographic region, gender, and study period. When only two groups were included in the subgroup analyses, differences between subgroups were evaluated using Cochran’s Q test (χ² test). When three or more subgroups were present, pairwise comparisons were performed to determine statistically significant differences between any two subgroups. We performed sensitivity analysis by sequentially excluding each study to assess its influence on the overall estimate. Publication bias was evaluated using funnel plots and Egger’s regression test, with p < 0.05 considered statistically significant.

Ethical approval

Not needed for this study.

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Results

Selection and characteristics of the study

A total of 189 studies were initially identified through database searches. After removing duplicates and screening titles and abstracts, 82 studies were selected for full-text evaluation. Of these, 17 met the inclusion criteria and were included in the meta-analysis (Fig. 1). The included studies, conducted between 2009 and 2022, covered 14 provinces across China, encompassing diverse climatic and ecological regions (Fig. 2).

Fig. 1. PRISMA flow diagram summarizing the study selection process.

Fig. 2. Geographic distribution of studies included in the meta-analysis of T. gondii infection in stray dogs across China. The map labeled the provinces covered by eligible studies and grouped by major geographic regions (North, Central, South, Northeast, Southwest, and Northwest China) using different colors. The lower-left panel summarizes region-specific pooled seroprevalence estimates derived from subgroup analyses. No data were available for the Southwest region because no eligible studies were identified.

This research incorporated studies detecting pathogen nucleic acids and seroepidemiological investigations of pathogen-specific antibodies (Table 1). Among them, 14 studies (n=2,112 samples) used serological methods (ELISA or IHA) to detect T. gondii antibodies, while 3 studies (n=208 samples) used PCR-based molecular assays to detect T. gondii DNA. The study sample sizes ranged from 17 to 918 dogs.

Pooled prevalence of T. gondii infection

The pooled seroprevalence of T. gondii in stray dogs was 31% (95% CI: 22%–40%) based on 2,112 samples from 14 studies (Fig. 3). Significant heterogeneity was observed among studies (I²=95%, p < 0.01). Three studies (n=208 samples) employed molecular techniques and reported DNA positivity rates ranging from 2% to 47% (Table 1); however, a pooled molecular prevalence was not calculated owing to the limited number of studies.

Table 1. Epidemiological studies on Toxoplasma gondii in stray dogs in China.

Fig. 3. Forest plot of the individual and pooled seroprevalence of T. gondii among stray dogs in China. Squares represent study-specific prevalence estimates; horizontal lines indicate 95% CI, and the diamond represents the pooled estimate from a random-effects model.

Table 2. Subgroup analysis of T. gondii seroprevalence in Chinese stray dogs.

Subgroup analysis

No subgroup analysis was conducted for T. gondii DNA detection. Notably, a study from Guangdong Province reported a positivity rate of approximately 47% (8 out of 17 samples positive), which was much higher than that of two studies from other regions (Table 1).

Subgroup analysis by sex showed a pooled seroprevalence of T. gondii of 27.3% (95% CI: 5.9%–56.4%) in males and 27.8% (95% CI: 6.2%–56.6%) in females, with no statistically significant difference between the two groups (Q=0, df=1, p=0.9773) (Table 2). Analysis by age group showed pooled seroprevalence estimates of 12.8% (95% CI: 0%–38.7%) in dogs aged 0–12 months, 38.3% (95% CI: 5.5%–78.1%) in those aged 13–36 months, and 35.7% (95% CI: 9.2%–67.3%) in dogs older than 36 months. Although the differences among age groups were not statistically significant (p=0.3582 for 0–12 m vs. 13–36 m; p=0.3308 for 0–12 m vs. >36 m; p=0.9228 for 13–36 m vs. >36 m), dogs aged >12 months tended to have higher seroprevalence.

Subgroup analysis based on geographic region and study period also showed no statistically significant differences among the subgroups (Table 2). However, stray dogs from southern provinces (e.g., Guangdong) exhibited relatively higher seroprevalence than those from northern and northeastern China (Fig. 2), a pattern also observed in the limited molecular data (Table 1). When stratified by diagnostic method, studies employing ELISA generally reported higher seroprevalence than those using IHA, and this differencea was statistically significant (Q=19.24, df=1, p < 0.0001) (Table 2).

Sensitivity analysis

The sensitivity analysis showed that the exclusion of individual studies did not substantially alter the overall effect estimate. Although a few outliers deviated slightly, all studies remained within the 95% CI, indicating that no single study had a disproportionate influence on the pooled results (Fig. 4).

Fig. 4. Sensitivity analysis of the pooled seroprevalence of T. gondii among stray dogs in China using a leave-one-out approach. Each row represents the recalculated pooled prevalence after omitting one study at a time. The diamond indicates the overall pooled estimate derived from the random-effects model, demonstrating the results’ stability and robustness.

Publication bias

Funnel plot analysis demonstrated an asymmetrical distribution of studies (Fig. 5), suggesting a potential publication bias. As the number of studies reporting T. gondii seroprevalence exceeded 10, the Egger’s test was conducted. The linear regression test indicated significant funnel plot asymmetry (t=2.23, df=12, p=0.0457; bias estimate: 4.1943, SE=1.8814), further supporting the presence of potential publication bias.

Fig. 5. Funnel plot of studies assessing potential publication bias in the seroprevalence of T. gondii among stray dogs in China. The x-axis and y-axis represent the estimated proportion and standard error, respectively. Each dot represents an individual study, and the dashed vertical line indicates the pooled estimate.

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Discussion

This systematic review and meta-analysis provide the first comprehensive synthesis of available evidence on T. gondii infection in stray dogs in China, highlighting their epidemiological relevance at the human-animal-environment interface.

The overall prevalence and epidemiological significance

The pooled seroprevalence observed in stray dogs was substantially higher than that previously reported in owned or pet dogs in China (Gao et al., 2016), supporting the notion that stray dogs experience greater cumulative exposure to T. gondii in the environment. Unlike owned dogs, stray dogs are continuously exposed to contaminated soil, garbage, prey animals, and livestock environment, which likely increases their probability of ingesting oocysts or tissue cysts. Therefore, the elevated seroprevalence likely reflects environmental contamination intensity rather than host susceptibility, reinforcing the value of stray dogs as sentinel animals for environmental contamination and T. gondii transmission dynamics.

Subgroup analyses by host factors, geographic location, and diagnostic methods

No significant differences in seroprevalence were observed across sex, age categories, region, or study periods. The absence of sex-related differences is consistent with the non-sex-specific exposure routes of T. gondii infection and has been widely reported in canine studies (Duan et al., 2012; Gao et al., 2016; Yang et al., 2024). Although older dogs exhibited relatively higher seropositivity, the lack of statistical significance suggests that the infection risk in stray dogs may accumulate rapidly early in life, reaching a plateau after that. This pattern contrasts with that observed in pet dogs, where age-related trends are often more pronounced (Gao et al., 2016), further emphasizing the high and continuous exposure pressure faced by stray dogs.

Geographic subgroup analysis did not reveal significant regional differences, despite higher point estimates in the southern provinces. This may reflect insufficient statistical power due to limited regional representation, but it may also indicate that environmental contamination with T. gondii is widespread across diverse climatic zones in China. The consistency between higher seroprevalence in southern regions and higher PCR positivity suggests that ecological factors, such as humidity, temperature, and livestock density, may influence parasite persistence, although current evidence remains insufficient for firm conclusions.

Notably, a significant methodological effect was observed between diagnostic assays, with studies using ELISA reporting higher seroprevalence than those employing IHA assays. This discrepancy may be explained in part by the smaller number of IHA-based studies and the generally higher analytical sensitivity of ELISA. Variability in antigen composition, cut-off thresholds, and assay standardization may also contribute to inconsistent estimates, as previously highlighted by (Dubey et al., 2020). Interestingly, a prior meta-analysis of T. gondii infection in pet dogs did not identify a comparable methodological difference (Gao et al., 2016), suggesting that assay-related heterogeneity may be more pronounced in stray dog studies, where sample quality and field conditions are more variable. These findings underscore the need for standardized serological protocols in future epidemiological investigations.

Limited molecular evidence and implications for surveillance

Only three studies employed PCR-based detection of T. gondii, reflecting the technical, biosafety, and cost-related barriers associated with molecular diagnostics. Limited molecular data constrain the interpretation of active infection and environmental shedding potential. Among the included studies, two using fecal samples reported markedly lower detection rates than the one analyzing blood samples. This likely reflects the transient nature of oocyst passage and DNA degradation in feces, whereas blood-based detection captures systemic infection. Although experimental studies have demonstrated that dogs may transiently excrete oocysts after ingesting large quantities (Lindsay et al., 1997), whether fecal PCR is inherently less sensitive than blood-based detection in natural settings remains unresolved. The scarcity of molecular data highlights the relative neglect of pathogen-focused surveillance in stray dogs, despite their potential epidemiological importance.

Public health implications of T. gondii infection in stray dogs

The public health relevance of these findings becomes clearer when stray dog data are compared with human and owned dog associated seroprevalence in China. National surveys estimate an overall seroprevalence of approximately 7%–8% in the general population (Zhou et al., 2011), with higher rates among pet dogs (~11%) (Gao et al., 2016). Against this backdrop, the pooled seroprevalence of 31% observed in stray dogs in the present study delineates a clear exposure gradient (stray dogs > pet dogs > general population), suggesting that stray dogs may serve as important sentinels of environmental contamination and may contribute to maintaining exposure pressure at the human–animal–environment interface, rather than mere incidental hosts. Through mechanical transport of oocysts on fur or paws, scavenging behavior, and close contact with contaminated environments, stray dogs may indirectly contribute to human exposure, particularly in densely populated urban settings. Despite this potential risk, toxoplasmosis remains under-prioritized within One Health frameworks in China, as reflected by the limited number of epidemiological studies focusing on stray dogs and the even scarcer application of molecular detection methods. This research gap likely underestimates the true contribution of stray dogs to environmental contamination and human exposure.

Limitations and future directions

Several limitations should be acknowledged. Substantial heterogeneity was observed as reflected by consistently high I² values, indicating considerable between-study variability and limiting the certainty of pooled prevalence estimates. This heterogeneity may partly arise from variations in diagnostic methods and inconsistent sampling frames, with some studies sampling shelter-based dogs and others sampling free-roaming stray dogs, thereby introducing ecological variation that could not be fully accounted for. Incomplete reporting of key host characteristics further constrained subgroup analyses. For example, among the 14 included seroprevalence studies, only three reported data stratified by age or sex, which limited the ability to assess these factors as potential heterogeneity sources. The limited number of molecular studies also restricted the precision of PCR-based prevalence estimates. Finally, publication bias may have contributed to the overrepresentation of studies reporting higher prevalence. Future research should prioritize standardized diagnostic approaches, expanded molecular surveillance, and genotyping of T. gondii isolates to better elucidate transmission pathways and public health relevance.

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Conclusion

This meta-analysis demonstrates that T. gondii infection is widespread among Chinese stray dogs. Future studies should prioritize standardized serological surveillance and emphasize molecular epidemiological investigations in stray dogs. To reduce environmental contamination and potential spillover risks, integrated One Health measures, including improved stray dog management, environmental sanitation, and public health education, are recommended.

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Acknowledgments

None.

Conflict of interest

The authors have no conflicts of interest to declare.

Funding

The Taicang Science and Technology Program (TC2021JC16) supported Chaoyang Chen. The sponsor was not involved in the implementation of the project or the writing of the paper. The views expressed are those of the authors and not necessarily those of the sponsor.

Authors' contributions

Chaoyang Chen: Conceptualization, protocol development, literature search, data curation, data analysis, software, and original manuscript drafting. Xiaoxia Ji: Conceptualization, protocol development, literature search, data curation, data analysis, software, manuscript drafting, and review. All authors have approved the final version of the manuscript.

Data availability

All relevant data are within the paper and its supporting information files.

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