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Background <p><i>Cyclospora cayetanensis</i> is an important foodborne parasite worldwide, with fresh produce and contaminated irrigation water as major transmission vehicles. In South Asia, environmental surveillance data remain limited. We investigated the occurrence of <i>C. cayetanensis</i> DNA in fresh produce and irrigation water across peri-urban areas of Khyber Pakhtunkhwa, Pakistan, and assessed environmental and farm-level factors associated with contamination.</p> Methods <p>A cross-sectional study was conducted in Peshawar and Kohat districts from April to September 2025. A total of 420 samples were collected, including 300 fresh produce samples (six commonly consumed vegetables and herbs) and 120 irrigation water samples from canal, tube-well, and mixed sources. Samples were processed using concentration techniques, and detection was performed by nested PCR targeting the 18&#xa0;S rRNA gene. Structured field questionnaires were used to capture farm-level practices, and logistic regression was applied to identify risk factors.</p> Results <p>We detected <i>C. cayetanensis</i> DNA in 6.0% of produce (18/300) and 12.5% of irrigation water (15/120; <i>p</i> = 0.028). Canal water (20.0%) was more frequently contaminated than tube-well water (5.0%; OR 4.75; 95% CI: 1.01–22.3). Leafy vegetables and herbs had higher contamination than smooth-surfaced produce (8.0% vs. 2.0%; <i>p</i> = 0.009). In multivariable analysis, canal irrigation (aOR 3.41), proximity to drainage channels ≤ 50&#xa0;m (aOR 3.98), and use of untreated rinsing water (aOR 2.91) remained independently associated with contamination (all <i>p</i> &lt; 0.05).</p> Conclusions <p>This study provides among the first molecular evidence of <i>C. cayetanensis</i> contamination at the produce–water interface in peri-urban Khyber Pakhtunkhwa, Pakistan, identifying surface irrigation and poor water management as key risk factors. However, because PCR detects DNA rather than viable organisms, these findings indicate environmental contamination and potential exposure pathways rather than direct infection risk. Sequencing confirmation is needed to exclude cross-amplification of related coccidia.</p>

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Molecular detection of Cyclospora cayetanensis in fresh produce and irrigation water in peri-urban settings: a cross-sectional study of the environmental-agricultural interface

  • 期刊论文

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  • 发表日期：

    2026年5月1日

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  • Iffat Naz,
  • Mina A. Almayouf,
  • Abdul Rehman,
  • Abdul Majid,
  • Abdul Basit,
  • Mubbashir Hussain

摘要

Background

Cyclospora cayetanensis is an important foodborne parasite worldwide, with fresh produce and contaminated irrigation water as major transmission vehicles. In South Asia, environmental surveillance data remain limited. We investigated the occurrence of C. cayetanensis DNA in fresh produce and irrigation water across peri-urban areas of Khyber Pakhtunkhwa, Pakistan, and assessed environmental and farm-level factors associated with contamination.

Methods

A cross-sectional study was conducted in Peshawar and Kohat districts from April to September 2025. A total of 420 samples were collected, including 300 fresh produce samples (six commonly consumed vegetables and herbs) and 120 irrigation water samples from canal, tube-well, and mixed sources. Samples were processed using concentration techniques, and detection was performed by nested PCR targeting the 18 S rRNA gene. Structured field questionnaires were used to capture farm-level practices, and logistic regression was applied to identify risk factors.

Results

We detected C. cayetanensis DNA in 6.0% of produce (18/300) and 12.5% of irrigation water (15/120; p = 0.028). Canal water (20.0%) was more frequently contaminated than tube-well water (5.0%; OR 4.75; 95% CI: 1.01–22.3). Leafy vegetables and herbs had higher contamination than smooth-surfaced produce (8.0% vs. 2.0%; p = 0.009). In multivariable analysis, canal irrigation (aOR 3.41), proximity to drainage channels ≤ 50 m (aOR 3.98), and use of untreated rinsing water (aOR 2.91) remained independently associated with contamination (all p < 0.05).

Conclusions

This study provides among the first molecular evidence of C. cayetanensis contamination at the produce–water interface in peri-urban Khyber Pakhtunkhwa, Pakistan, identifying surface irrigation and poor water management as key risk factors. However, because PCR detects DNA rather than viable organisms, these findings indicate environmental contamination and potential exposure pathways rather than direct infection risk. Sequencing confirmation is needed to exclude cross-amplification of related coccidia.