1.10: Laboratory

Learning Objectives

• After reading this chapter you should be able to identify and explain the following topics related to treatment plant laboratories:
• Equipment in the laboratory
• Laboratory safety
• Water quality testing
• Contaminants tested in the laboratory and throughout the treatment plant

Working in a laboratory is probably something you never thought about when deciding to explore the career of water treatment. However, working in a laboratory is one of the more critical aspects of the profession. Many larger agencies have a dedicated staff of laboratory personnel that handles a lot of the day to day water quality analysis, but operators still play a key role. After all, the operator is the person in charge of running the plant, monitoring chemical feeds, and performing routine plant checks to ensure the system is running smoothly. A laboratory scientist can run all the tests in the world, but since they are not the ones running the plant, a lower chlorine residual may not stand out to them. The responsibility falls squarely on the shoulders of the operator. State regulations require operators to take grab samples throughout the day to ensure monitoring equipment throughout the plant is running correctly.

This topic is discussed at length in the water quality course. This is meant to be a brief overview if you have not taken that course yet.

Laboratory Instruments and Equipment

Throughout the lab, you will find a variety of beakers, flasks, dilution bottles, and graduated cylinders. A lot of these instruments look the same or similar but they all have a unique job and purpose. Beakers range in size from 25 to 4000 mL. They are used to mix samples during chemical analysis. A burette is a long skinny tube-like glass receptacle used to measure and disperse liquid. Flasks have a narrow neck and are round at the bottom. Each kind of flask serves a different purpose, but they all look very similar.

Figure \(\PageIndex{1}\): (left) Beakers – Image by Xavax is in the public domain. (right) Volumetric flask – image by Lucasbosch is licensed under CC BY-SA 3.0

Graduated cylinders are tall and cylindrical and range in size from 10 mL-4000 mL. They are used to measure liquid, but not with accuracy. For example, they can be used during a chlorine test to gather a general volume of water. If you need an exact amount, it should be measured with a pipette or other similar and accurate liquid measuring device. The pipette measures a liquid with 100 percent accuracy.

Figure \(\PageIndex{2}\): Sample Bottle – Image by Σ64 is licensed under CC BY-SA 3.0

You will find sample bottles in any laboratory. The bottles can be made of plastic or glass. They are used to store water for future lab tests and not used for measuring liquid. Sample bottles are used to collect bacteriological samples and for organic chemical analysis. The bacteriological bottle is one of the most important sample bottles you find in a lab. They are made of plastic and are 100 mL. Water treatment operators must take bacteriological samples per the Total Coliform Rule. These tests will be performed daily in the lab and throughout the distribution system based on your coliform sampling plan.

Figure \(\PageIndex{3}\): (left) Graduated Cylinder – Image by Lilly_M is licensed under CC BY-SA 3.0. (right)  Burette – Image by Roland1952 is licensed under CC BY-SA 3.0.

Other Items Found In the Laboratory

Incubators will be found in every lab at a water treatment plant. They are used to hold a temperature for bacteriological cultures. There are two types of incubators used, dry heat and water baths. Common uses for incubators include coliform testing, multiple tube testing, and heterotrophic plate counts. Ovens are other pieces of equipment found in the lab. Autoclaves, for example, are used to sterilize glassware items but can also be used to sterilize material that has been contaminated from perhaps a positive coliform test. Refrigerators are used to store samples for future testing and to store chemicals used for water quality testing. These refrigerators are for work use only and should not be used to store food and beverages.

Laboratory Safety

Operators should always be aware of safety regardless of where they are at the treatment plant. The laboratory should be treated no differently than if an operator was changing chlorine cylinders. There are chemicals that are combustible and others that can cause severe burns if they are exposed to skin. Some common safety equipment found in water treatment plant laboratories:

• Eye protection⁠—Eye protection should include safety goggles, safety glasses, or face shields. It is not recommended to wear contact lenses if working with dangerous chemicals. Prescription safety eyewear is available. You only have one set of eyes, so it is important to protect them. Water laboratories have liquids and solids that can easily penetrate your eyes if you are not wearing the proper PPE (personal protective equipment).
• Safety Showers/Eyewash Bottles⁠—In the event that something gets in your eyes, every lab is equipped with a safety shower. The shower should have a pull lever and an eyewash sink.
• Fire Extinguisher⁠—Fire extinguishers are located throughout any building in the United States. Most water treatment companies will provide formal training on proper use of a fire extinguisher. In the event that you cannot put a lab fire out with the use of an extinguisher, exit the room right away and call the fire department. Fire extinguishers are meant to put out very small fires very quickly.

Figure \(\PageIndex{4}\): Emergency eyewash and shower – Image by Korn966 is licensed under CC BY-SA 4.0

Meters

Laboratories use a variety of meters for water quality testing. Although meters are located throughout the treatment plant, the meters in the lab are used to verify that equipment throughout the treatment plant is functioning correctly. The most common meters used by water treatment operators are pocket colorimeters for chlorine testing in the field, turbidity meters, and pH meters. Pocket colorimeters are great to be used in the field, but the results are not reportable. For compliance purposes, you must use an electric colorimeter or photometer.

pH meters and turbidimeters are used to verify equipment is working throughout the plant. At a minimum, an operator will run labs on turbidity, chlorine residual, water temperature, and pH. All of these parameters are required for CT compliance and verification. These measurements are important in ensuring the treatment plant is operating in compliance with the surface water treatment rule.

Microbiological Testing

Microbiological testing is probably the most important testing done by water treatment professionals. As discussed at length in this text, treatment water operators aim to remove and deactivate pathogenic organisms from drinking water through filtration and disinfection. Water regulations require 3Log removal and deactivation of Giardia and 4 log removal and deactivation of viruses. Bacteria fall in the middle of Giardia and viruses. Therefore, it is safely assumed bacteria will be removed and deactivated along the same process.

Water professionals test for microbiological agents by testing for the indicator organism coliform. Coliform bacteria cause no harm to humans but are generally present when pathogens are present in the water. The reason coliforms have been used to identify contaminated water for over 100 years is because they are always present in contaminated water. Even if there is no fecal contamination, they are still present. They survive longer in water than pathogens and they are easy to identify with proper testing. When a positive sample is identified, we assume the water is contaminated until it can be proven otherwise.

Water treatment facilities use four different tests for coliform monitoring. The easiest, least expensive, and the most common method for coliform testing is the presence absence test (P-A). Both the P-A and multiple tube fermentation (MTF) tests work based on the fact that coliform produces gas from the fermentation of lactose within 24-48 hours. The MTF method uses three steps for the test. It includes the presumptive, confirmed, and completed test cycle. The presumptive test is accomplished in 24 hours. Samples are incubated at 35 degrees Celsius for 24 hours and then checked to see if a gas bubble has formed or if the sample is cloudy. You want to see no bubble or gas. The samples are then incubated for another 24 hours. If gas does not form, the test is over and the sample is absent. If there is a gas bubble, you move on to the confirmed test.

The confirmed test verifies the sample is positive from coliform and not another type of bacteria. Brilliant Green Lactose Vile is added to the sample and then incubated for another 48 hours. The same method is used. The lab technician or operator checks to see if gas is produced during the incubation period. The minimum requirement for water treatment operations is the presumptive and confirmed test. The completed tests are rarely used except for quality control by laboratory personnel. In the event of a positive confirmed test, coliform bacteria violation protocol will go into effect.

The P-A method is the most common method for treatment operators and field sampling staff. The bottles are easily transferred in an ice chest and the set-up is very simple. This method is commonly referred to as the Colilert method. The test uses a 100 mL plastic sample bottle. A nutrient is added to the sample, which is then incubated for a period of 24 hours at 35 degrees Celsius. The nutrient will cause the water to turn yellow if coliforms are present. In the event of a positive test, the bottle can be placed behind a fluorescent light that will turn a blue color which indicates fecal E. coli contamination.

The other two bacteriological tests include MMO-MUG and the membrane filter method. The MMO-MUG tests come with the testing agent already in the vials. A 10 mL sample is simply added to the vial. As with the P-A test, samples positive for coliform will turn yellow and for fecal contamination will turn blue. The membrane process begins with filtering 100 mL of water through a membrane filter. Then the sample is added to a petri dish and incubated for a 24-hour period. Coliform positive samples turn a red-pink color. Confirmative tests require incubation of a broth for another 24 hours.

Physical Water Quality

Operators test the water for both physical properties and contaminants that will cause harm to humans. There are secondary standards related to many of the physical tests operators perform. Acidity is the ability to neutralize a base. Alkalinity is the ability of water to neutralize acid. The reason operators are concerned with the acidity and alkalinity of drinking water is different pH scales have different effects on water treatment. When acids are added to water, they lower the pH of the water. A strong base may need to be added to boost the pH. Operators want a small amount of calcium carbonate present in the water to deposit around the pipes of the distribution system. This helps combat corrosion.

When we think of water drinkability, we might think of color, taste, odor, and temperature. The physical aesthetic of the water matters and impacts our desire to drink the water. The color of the drinking water may indicate water with higher levels of organic compounds and water with THMs.

Taste and odor issues in treated drinking water are hard to test. Many customers complain of a strong chlorine smell in their drinking water. Taste and odor problems can come from organic matter, chlorine, dissolved gases, and even industrial wastes. The threshold odor test is measured on the TON scale. (Water sample of 100 mL + dilution divided by 100 = TON.) Water registering as a 3 will most likely draw complaints from customers.

Water temperature plays a key role in water treatment because disinfectants work better at higher temperatures. However, nitrification occurs during the warmer months of the year so the water temperature has to be carefully managed. Water agencies in colder regions of the country must deal with freezing conditions within their treatment and distribution system. During the winter even in warmer regions, lake turnover can cause vast changes in water quality.

Turbidity is the suspension of particles in water. High levels of turbidity can signify major issues within the treatment plant. Therefore, turbidity is used to verify how efficiently the treatment plant is operating. High turbidity values may indicate higher levels of organic and inorganic matter. Pathogenic organisms can hide behind turbidity and render disinfection ineffective. Turbidity is measured in the lab with a nephelometric counter. A light source is added and, in the presence of turbidity, the light scatters, producing higher numbers with greater amounts of turbidity.

The final two water quality testing procedures most commonly performed by water treatment operators are chlorine and pH. The chlorine or chloramine feed system is one of the most critical components of a water treatment plant that operators monitor through online testing and grab samples.

The measurement of the hydrogen ion concentration in water is pH. The pH scale ranges from 1- 14 with 1 being the most acidic and 14 the most basic. The higher ends of both ranges produce the most corrosiveness. 7 is considered neutral. Do not confuse pH with acidity or alkalinity. It is important to monitor pH because the pH is used to control many chemical reactions in the treatment plant including coagulation, disinfection, corrosion control, and the removal of ammonia. pH also plays a key role in the CT calculation.

Chapter Review

1. An organism used to indicate the possible presence of E. coli contamination is ___________.
   1. Cryptosporidium
   2. Giardia
   3. Coliform
   4. Brilliant green vile
2. The presence-absence (P-A) test used for microbiological testing is also commonly referred to as ___________.
   1. Multiple tube fermentation
   2. Membrane filtration
   3. Confirmed test
   4. Colilert
3. When testing for coliform bacteria with the multiple tube fermentation (MFT) method what is the best indicator for a positive test?
   1. Color change
   2. Gas bubble formation
   3. Formation of a cyst
   4. Formation of turbidity
4. Coliform bacteria share many characteristics with pathogenic organisms. Which of the following is not true?
   1. They survive longer in water
   2. They grow in the intestines
   3. There are less coliform than pathogenic organisms
   4. They are still present in water without fecal contamination
5. What is the second step in the multiple tube fermentation test?
   1. Presumptive test
   2. Negative test
   3. Completed
   4. Confirmed
6. What is the removal and deactivation requirement for Giardia?
   1. 2 Log
   2. 3 Log
   3. 4 Log
   4. There is no requirement
7. The multiple barrier approach to water treatment includes removal through which method?
   1. Filtration
   2. Coagulation
   3. Disinfection
   4. Both 1 and 3
8. A pH reading of 7 is considered ___________.
   1. Slightly acidic
   2. Acidic
   3. Basic
   4. Neutral
9. A higher than normal turbidity reading could signify ___________.
   1. A change in water quality
   2. Nothing. Keep operating as normal
   3. Microbiological contamination
   4. Both 1 and 3
10. What is the ingredient used during the second multiple tube fermentation test?
    1. Colilert
    2. MMO/MUG
    3. Brilliant green vile
    4. Chlorine