Methods 1623 and 1623.1 Technical Training Material

• Are provided as guidance for those interested in technical training for EPA Methods 1623 and 1623.1.
• Were developed for the implementation of the Long Term 2 (LT2) Enhanced Surface Water Treatment Rule.
• Cover sample collection and handling, technical instruction for laboratories using EPA Methods 1623 and 1623.1, and training materials for individuals who will audit laboratories analyzing water samples for Cryptosporidium or Giardia using these methods.
• Could be of interest to water system employees, laboratories, or individuals who will audit laboratories running these methods.

• Training for Cryptosporidium sample collection
• Training for laboratories analyzing water samples using EPA Methods 1623 and 1623.1
• Training for auditors of laboratories using EPA Methods 1623 and 1623.1

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Training for Cryptosporidium sample collection

The following training materials describe best practices and offer guidance for individuals collecting surface water samples that will be analyzed for Cryptosporidium or Giardia using U.S. EPA Methods 1623 and 1623.1.

Disclaimer: Viewers should expect some long periods of audio silence as these training videos play, allowing time to carefully read the presented material.

• LT2 Rule Cryptosporidium and E. coli Sample Collection Recommendations (YouTube) (Run Time 33.06 minutes)
  This video details procedures for bulk sample collection, field filtration and instruction on proper sample packing and shipping. Supplemental material:
  • LT2 Rule Cryptosporidium & E. coli Sample Collection Recommendations - Pocket Guide (pdf) (1.1 MB)

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Training for laboratories analyzing water samples using EPA Methods 1623 and 1623.1

• Overview of US EPA Cryptosporidium and Giardia Detection Methods (YouTube) (Run Time 10.7 minutes)
  This video provides an overview of the methods developed by the EPA for Cryptosporidium and Giardia detection in water. The methods described in this video are:
  • Method 1623.1: Cryptosporidium and Giardia in Water by Filtration/IMS/FA January 2012 (pdf) (1 MB)
  • Method 1623: Cryptosporidium and Giardia in Water by Filtration/IMS/FA December 2005 (pdf) (1.3 MB)
  • Method 1622: Cryptosporidium in Water by Filtration/IMS/FA December 2005 (pdf) (1.4 MB)
• Cryptosporidium Analysis: Using a Dispersant (YouTube) (Run Time 2.4 minutes)
  This video demonstrates the dispersant addition procedure, detailed in EPA Method 1623.1.
• Immunomagnetic Separation (YouTube) (Run Time 32.27 minutes)
  This video details the immunomagnetic separation (IMS) procedure used in EPA Method 1623 to separate Cryptosporidium oocysts and Giardia cysts from extraneous debris in a water sample. Supplemental material:
  • Method 1623 IMS Procedure-Supplemental Information (pdf) (426 k)
• Basic Microscopy (YouTube) (Run Time 30.05 minutes)
  This video assists analysts in the proper use and maintenance of a microscope as used to detect Cryptosporidium and Giardia in Method 1623.
• Microscopic Identification (YouTube) (Run Time 29.29 minutes)
  This video helps analysts properly identify and characterize Cryptosporidium and Giardia using Method 1623. The video includes characterization by FITC, DAPI and DIC of positive control organisms and interfering organisms.
• Cryptosporidium in Water: Reviewing Laboratory Reports (YouTube) (Run Time 2.5 minutes)
  This video describes key review elements to increase one's understanding of laboratory reports and quality control for Cryptosporidium analyses.

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Training for auditors of laboratories using EPA Methods 1623 and 1623.1

The following videos will provide detailed technical information about the EPA Methods 1623 and 1623.1 and how to best evaluate the laboratory’s performance. This video course is divided into 12 different videos. The videos are best viewed in numerical order as material in the first videos is referenced in later videos. These videos are meant to be used in conjunction with the Manual for the Certification of Laboratories Analyzing Drinking Water, Fifth Edition, Supplement 2 (EPA 815-F-12-006), which can be found at:

• Supplement 2 to the Fifth Edition of the Manual for the Certification of Laboratories Analyzing Drinking Water (pdf) (877 k)

1. Requirements for Cryptosporidium Monitoring for the Long Term 2 Enhanced Surface Water Treatment Rule (YouTube) (Run Time 16.39 minutes)
   This video describes the requirements of the Long Term 2 Enhanced Surface Water Treatment (LT2) Rule as they apply to the laboratories. 
2. Introduction of Cryptosporidium and Giardia (YouTube) (Run Time 10.40 minutes) 
   This video provides material about Cryptosporidium and Giardia, and the different methods to detect them. This information will help acquire a basic knowledge to audit a Cryptosporidium and Giardia laboratory as well as address some of the hazards associated with visiting these laboratories. 
3. Personnel, Equipment & Reagents (YouTube) (Run Time 9.25 minutes)
   This video introduces some of the requirements of the staff, equipment, and reagents within a laboratory analyzing for Cryptosporidium and Giardia.
4. Sample Collection and Spiking Quality Controls (YouTube) (Run Time 10.23 minutes)
   This video covers the collection and acceptance of samples, as well as the fortification, or spiking, of quality control samples when using EPA Method 1623.1.
5. Filter Elution and Sample Concentration (YouTube) (Run Time 20.08 minutes)
   This video covers the steps to elute the collected sample from the filter as described in EPA Method 1623.1. The elution procedures for two of the filters, Envirochek HV and Filta-Max, are described.
6. Immuno-magnetic Separation (YouTube) (Run Time 6.30 minutes)
   This video covers immuno-magnetic separation, or IMS. The purpose of IMS is to isolate as many Cryptosporidium oocysts and Giardia cysts as possible from the other particles within a source water sample.
7. Sample Staining (YouTube) (Run Time 4.30 minutes)
   This video covers sample staining. The goal of sample staining is to brightly highlight the Cryptosporidium oocysts and Giardia cysts as fluorescent green so they can be detected during the microscopic examination.
8. Microscope Inspection (YouTube) (Run Time 7.08 minutes)
   This video describes the critical parts of the microscope and what to watch when an analyst is demonstrating their skills on the microscope.
9. Evaluation of Analyst Performance (YouTube) (Run Time 6.03 minutes)
   This video covers the final step of EPA Method 1623.1 which is to examine the slide for organisms. This video will teach an auditor what to notice when an analyst is highlighting organisms to demonstrate their skills.
10. Quality Control Records (YouTube) (Run Time 10.06 minutes)
    This video outlines an approach for examining quality control records for Methods 1622, 1623 and 1623.1. It emphasizes the foremost parameters in a Cryptosporidium laboratory: holding times, ongoing precision and recovery and complete records for examining positive results in a field sample.
11. Documenting Laboratory Performance (YouTube) (Run Time 6.07)
    This video outlines how Cryptosporidium laboratories document their performance. Each lab is required to operate a formal quality assurance program that addresses key elements such as: data quality, equipment maintenance, reagent performance, analyst training, and records storage.
12. Unique Elements and Tools for the Cryptosporidium Audit (YouTube) (Run Time 5.56 minutes)
    This video outlines tools and suggestions to help an auditor optimize their time assessing a laboratory using EPA Methods 16231 and 623.1. These tools include items to request from the laboratory and a plan to evaluate the slide preparation and the analyst’s parasite identification skills.