Human pathogenic microsporidia detection in agricultural samples: method development and assessment.

Area of Science:

• Veterinary Microbiology
• Environmental Science
• Molecular Diagnostics

Background:

• Microsporidia are opportunistic pathogens that can affect livestock and humans.
• Accurate detection of microsporidia in environmental and fecal samples is crucial for disease control.
• Existing methods may lack sensitivity or specificity for diverse sample types.

Purpose of the Study:

• To develop and validate a sensitive method for recovering and identifying microsporidia from livestock fecal and environmental samples.
• To assess the detection limits of the method using spiked samples.
• To screen environmental samples for the presence of pathogenic microsporidia.

Main Methods:

• Sample purification via sucrose flotation.
• DNA extraction using the Qiagen QIAamp DNA Stool Mini Kit.
• Polymerase chain reaction (PCR) with generic microsporidia primers, followed by DNA sequencing.

Main Results:

• The method demonstrated sensitive recovery of Encephalitozoon intestinalis spores from cattle and swine wastewater and cattle feces.
• Detection was successful at spore concentrations as low as 100 spores/ml in swine wastewater.
• Two of 34 environmental samples tested positive for human pathogenic microsporidia.
• Encephalitozoon cuniculi was detected in swine feces and wastewater for the first time.

Conclusions:

• The developed method provides sensitive and specific detection of microsporidia in challenging sample matrices.
• This technique is valuable for monitoring microsporidia in livestock populations and their environment.
• The findings highlight the potential for zoonotic transmission and the need for further epidemiological studies.