Microbiological Methods for Examination of Water & Wastewater

Upon successful completion of this course, the delegates will be able to:

• Examine wastewater for biological characteristics, identify various organisms, perform particle size and number counts, and advise the operators of a wastewater treatment plant on the implications of the microbiological information developed
• Understand how the changes in microbiological populations in a wastewater treatment plant can affect the plant performance and what to do to bring it back into the proper operation
• Participants will also be able to use JNOPTIC Capture 2.2 software

Agenda

Day 1

• Introduction to Microscopy
• Basic Microscopy
• Introduction to Microscopy
• Different types of Microscopy
• Bright Field Microscopy
• Cross-Polarized Light Microscopy
• Dark Field vs. Bright Field
• Variants on Dark Field Microscopy
• Examples of Rheinberg Darkfield Microscopy
• Kohler Illumination Microscopy
• EPI Illumination
• Phase Contrast Microscopy Theory and Practice

Day 2

• Basics Taxonomy
• Order of Biology
• Kingdom-Phylum- Class –Order –Family –Genus -Species
• Types of Microorganisms:

Bacteria

Protozoa: (including Guardia and Cryptosporidium)

Metazoa: (Multicellular organisms)

Viruses

Important Terms

Parasites and Pathogens: worms and fungi

Soil Microbes and Heterotrophs: Bacteria; protozoans; nematodes; Obligate aerobes; Anaerobes; facultative organisms

• Bacteria Characteristics and Identification of types

• Size & Shapes Sheathed; Stalked; Budding; Filamentous; Cyanobateria

• Parts of the Cell and Identification

• Introduction to Dye Staining

• Gram Positive vs Gram Negative

• Dye staining techniques

• Gram Negative Bacteria Flowchart

• Components of a bacterial cell

• Extracellular proteins

• Flagella and Pili Motile bacteria

• Inclusions and vacuoles

• Endospores

• Autotrophs Heterotrophs

• Photoautotrophs

• Chemolithrophs

• Aerobic and Anaerobic Bacteria: Psychrophilic; Mesophilic and Thermophilic Bacteria Typical Growth Patterns; Time and concentration for growth

• Protozoa Identification stramennopiles protozoa, fungi and plants

• Amoebae: Ciliates, Common Wastewater Ciliates, Rotifers, Wastewater Rotifers

• Rotifer Identification

• Nematodes & Roundworms

• Using a Counting Chamber Hemocyometer

• Other Metezoa Water bears and Water Fleas

• Helminth Ova:

Some associated diseases

Day 3

Wastewater Microbiology Applications

The Activated Sludge Process

Detailed Walk through on local Wastewater Treatment Plant

• Organotrophs aerobic or anaerobic, includes all animals and fungi, and many bacteria and protists.

Organotrophs' energy extraction

Wastewater Microbiology Applications Nitrifying bacteria

• chemoautotrophs
• chemolithotrophs:

Nitrosomonas, Nitrosococcus, Nitrobacter, Nitrococcus and their significance

Wastewater Microbiology Applications

Nitrification and denitrification

Population Dynamics

Day 4

Detailed Practices

FLAVOBACTERIUM

MICROBIOLOGICAL FLOC FORMATION

Types of problems:

• Pinfloc
• Filamentous Floc & Bulking
• Rising Floc
• Foaming

Factor affecting floc formation:

• Clarifier Underflow/Sludge Wasting

Filamentous Organism and Wastewater Treatment

• Tests and Procedures on Mixed Liquor in an AS System
• Measuring Total Floc Length

Identification of filaments & Jenkins guide

Nocardia spp.Type 1701

Type 021N

Type 0041

Thiothrix spp.

Sphaerotilus natans

Type 0092

Haliscomenobacter hydrossis

Type 0675

Factors affecting growth of Filamentous Organisms:

• MCRT
• DO Concentration
• Toxics (including chlorine)
• pH
• Sulfide

Filament Control methods

• Selective Toxicity
• Chlorination control
• Hydrogen Peroxide
• Ozone
• nutrient additions vs. deficiencies
• compounds to add
• Foam and Scum Controls

Identification of Filamentous Bacteria

A Simple Approach

What’s Needed A listing of supplies

• Separation by Staining
• Graham and Neisser
• Typical Observations Filament Shape
• Filament Size
• Cell shape
• Cell size
• Cell Septa
• Indentations
• Sheath
• Branching and other properties

Beggiatoa

More on bulking and foaming and how to identify various organisms

Nocardia

Good Practice

• Keep a process chart of treatment system parameters
• Measure the parameters consistently and routinely
• Measure the parameters when the system is running properly
• Exercises and Class Problems

Day 5

Software & Applications:

• Helminth Ova,
• significance, & control: The art of microscopic photography, Counting Chamber use;

Software use

Lab Work