Analyses of at least 5 representative batches of production or pilote-scale batches is necessary to determine the content of the pure active substance, microbial contaminants and pathogens.The manufacturing processes have the potential of producing unwanted micro-organisms in addition to the desired  microbial pest control agent (MPCA) which include pathogens, their associated toxins and other metabolic by-products of health concern.

Contaminants and pathogens for Bacteria based technical materials/formulated products:
– Viable yeasts and moulds
– Anaerobic spore formers
– Pseudomonas aeruginosa
– Coliforms or Escherichia coli
– Staphylococcus aureus
– Listeria monocytogenes
– Vibrio cholerae
– Shigella spp.
– Salmonella spp.

Contaminants and pathogens for Fungi based technical materials/formulated products:
– Aerobic plate count
– Anaerobic spore formers
– Pseudomonas aeruginosa
– Coliforms or Escherichia coli
– Staphylococcus aureus
– Listeria monocytogenes
– Shigella spp.
– Salmonella spp.

STUDY DESIGN
Analysis of the 5 batches, in terms of determination of the active ingredient content,  are carried out by an analytical method validated according to SANCO/3030/99 rev. 4  (11/07/2000) on 1 of the 5 batches

Determination of the active substance*
3 samples  (as 3 independent weighings) of each batch
Progressive dilutions, for each of the 3 samples, until reaching the appropriate dilution factors

Quantification/detection of microbial contaminants and pathogens*
1 sample of each batch
Progressive dilutions of the sample until reaching the appropriate dilution factors or sample enrichment

* Additional determinations, upon Sponsor’s request, can be carry out on one batch within a shelf life study

LIMITS
Viable yeasts and moulds: < 10³ CFU/g or mL
Anaerobic spore formers: < 10⁵ CFU/g or mL
Aerobic plate counts: < 10⁵ CFU/g or mL
Listeria monocytogenes: Absence in 25 g/25 mL
Shigella spp.: Absence in 25 g/25 mL
Pseudomonas aeruginosa: Absence in 1 g/1 mL
Salmonella spp.: Absence in 25 g/25 mL
Vibrio cholerae: < 1 CFU/ 25 g
Coliforms: < 10 CFU/g or mL or E. coli:  Absence in 1 g/1 mL
Staphylococcus aureus: Absence in 1 g/1 mL

Study includes GLP managment and reporting (if required)

REFERENCES AND GUIDELINES
SANCO/12116/2012 rev.0 (September 2012) and ISO methods