Wastewater Microbiology Bob Rawson Recommended Text Books Manual

Wastewater Microbiology Bob Rawson

Recommended Text Books • Manual of instruction for Wastewater Treatment Plant Operators two Volume New York State Department of Environmental Conservation LC control OL 220555 M. • Operation of Wastewater Treatment Plants Ken Kerri. • Wastewater Engineering Treatment Disposal /Reuse. Metcalf & Eddy Dr. George Tchobanoglous, U. C. Davis. • Sewerage Organisms a Color Atlas, J Cox, Chicago.

Recommended References continued • Standard Methods for the Examination of Water and Wastewater, 21 st Edition or most recent available. • MOP 11. WEF. • EPA Technology Transfer see internet web pages. • White’s Manual of Chlorination. • Any good recent Microbiology text book

Topics we will discuss • Structure of the Cell. Plant Animal Bacterial. • Fixed Film and Suspended Growth. (Examples) • Biological Mechanisms that affect water Quality and accomplish WW Treatment like: – Photosynthesis – Respiration Aerobic, Anaerobic and Facultative. – Nutrient Assimilation, importance & Cycles of • Carbon, Nitrogen, [phosphorous, sulfur.

Discussion Topics continued • Description of Wastewater Microbes. Structures Pathogens: Disease causing organism Microorganisms used in Wastewater Stabalization Obnoxious organisms causeing Odor, Corrosion toxicity. • Use of the Microscope. • Common Laboratory test methods. • TSS • Coliform Tests • BOD • Bioasays

Three Domains Six Kingdoms • Bacteria (Monera) • Archea • Eucaryota – Plants – Animals – Fungi – Protista (The Protozoans) • Protophyta • Protozoa • Protomycota

Phylogenetic Tree of Life

Taxonomy • Domain – Kingdom – Phylum (Division) – Subphylum – Class – Order – Family Binomial Genus Species

Example of Taxonomy • • • Eucaryota Kingdom Animalia Phylum Chordata Subphylum Terrapoda Class Mammalia Order Carnivora Family Canidae Genus Canis Species Familiarias (The Familiar Dog)

Microbial Taxonomy Naming, classifying microorganisms • 1. Naming -Linnaeus introduced binomial nomenclature • -Every organism is given a unique two-part name • Genus + specific epithet = species name • Ex. Escherichia coli, Yersinia pestis, Bacillus anthracis • 2. Classification -Organisms with similar characteristics are grouped together • Ex. Genus Bacillus • Bacillus anthracis, Bacillus subtilus, Bacillus cerus

Microbial Taxonomy Naming, classifying microorganisms • -Higher levels of organization are used to group species having similar characteristics • Ex. Bacillaceae is the name of a Family that contains several different species of rod-shaped, spore-forming, Gram positive, aerobic bacteria such as Bacillus anthracis, Bacillus subtilus and Bacillus cereus • 3. Identification • -Determination of the name of an organism that has been isolated by observing its characteristics

Differences Between Plant and Animal Cells. • Plant Cell • Rigid support and Protection • Cellulose a fiberous complex polysaccharide allows H 2 o and air to pass through. • Lignin a hard phenolic compound. Will react with chlorine or fire to form dioxin. • Plastids: • leucoplasts Starch

Plant Cell continued • Chromoplasts like Chloroplasts • Chloroplasts contain Chlorophyll • Responsible for photosynthesis by acting as a catalyst to split H 2 O in the presence of light. • We will discuss Photosynthesis further.

Plant Cell

Animal Cell

Typical Animal cell • • Cell Wall not absence of cell wall. Cytoplasm. The intercellular contents Organelles including: Plastids. Mitochondria. The power house ATP transactions. Golgi Complex. Stores enzymesd Cilia or flagella. Movement Food Vacuole

Microorganisms and Their Impacts • Microorganisms were the first living organisms on the planet – Prokaryotes (bacteria) 1 st appeared ~3. 5 billion years ago • Can be found nearly everywhere on earth in large numbers • -Soil, oceans, deep underground, extreme environments, on and in higher organisms

• ~100 trillion (1014) microorganisms person • 10 X number of cells of the human body ! • Most are in the intestinal tract • ~1 trillion bacteria on the skin • Normal flora aids food digestion and synthesizes vitamins in intestinal tract, inhibit growth of pathogenic microorgansisms by competiting for space and nutrients

Importance of Microbes • Major affect on the composition of gases in earth's atmosphere – -Formation of oxygen by photosynthesis (required for survival by many organisms) First source of Oxygen was Cyanobacteria. – -Formation of carbon dioxide and methane during decomposition (Greenhouse gasses) • Transformations of nitrogen (major plant nutrient) • Fix Carbon and Decompose organic matter. – Agriculture, Wastewater Treatment, Bioremediation Phytoremediation and Mycoremediation. • Recycle nutrients – -Carbon, nitrogen, phosphorus, sulfur • Microorganisms also cause diseases. Pathogenic organisms

-There are two main types of microbial cells • 1. Prokaryotic • 2. Eukaryotic • • • 1. Prokaryotic Small Lack a nucleus or organelles Often exist as single cells (unicellular) All bacteria are prokaryotes 2. Eukaryotic Larger than prokaryotes Have a nucleus and organelles May be unicellular or multicellular Yeast, fungi, protozoa, algae, helminths

Main types of microbial cells Cont. • • • 2. Eukaryotic Larger than prokaryotes Have a nucleus and organelles May be unicellular or multicellular Yeast, fungi, protozoa, algae, helminths

Typical Animal Cell Continued • Endoplasmic Reticulum with nodular lysosomes. • Lysosomes containing digestive enzymes

Virus • -Viruses are small particles composed of protein and nucleic acid (DNA or RNA) • Do not have the characteristics that define cellular organisms (see below) • Only able to reproduce when inside a cell • Obligate parasites that may have arisen as a degenerate form of early bacteria.

Bacteria Cell

Bacteria Cell • Capsule or Slime layer • Slimes are Sticky Gelatinous Material secreted by bacteria Cell • Functions of slime layer. • Allow bacteria to stick together onto substrate or flocculate • Protection from toxic environment like chlorine UV

Further functions of Slime Layer • Bacteria Stick together to a substrate or floculate. • Protection from hostile environment • Trap Food which is adsorbed to the cell surface by: – Adsorb =Adhere to surface, in preparation for absorption. • Slimes contain enzymes which Digest food. • Slimes contain endotoxins and exotoxins that cause disease reactions.

Sizes of Microorganisms • The micrometer, mm, is the unit of measure often used to describe the size of microorganisms • 1 mm = 1/1, 000 meter = 0. 000001 m = 1 x 10 -6 m • Typical sizes • Viruses 0. 01 to 0. 2 mm • Bacteria 0. 2 to 5 mm • Yeast 5 to 10 mm • Algae 10 to 100 mm • Protozoa 50 to 1000 mm

Activated Sludge • A suspended Growth treatment Process using microorganisms including; bacteria; fungi; and protozoans. • Three steps: – Transfer of food – Conversion of food to Cell Mass – Flocculation or settling of organisms that are stuck together.

Activate Sludge Transfer Continued • Adsorption: Particles and colloids stick to the slime layer • Absorption: Nutrients passing through the cell wall. • Phagocytosis/ Pinocytosis • Digestion: • Osmosis solute moves from more concentrated location to less concentrated location down gradient.

Transfer continued • Dialysis. Opposite of osmosis requires energy • Active Transport. Water and oil don’t mix until we add soap, or an emulsifier that acts as a carrier. • Cell membranes contain phospholipid • Large molecules that are on the outside like glucose cannot get in like small uncharged particles. To get in a carrier molecule attaches to the particle making it soluble in the lipid layer just like soap makes oil soluble in water. This requires energy provided by ATP.

Further steps in Activated Sludge Process • Second Step is the Conversion processes: • Respiration. – Aerobic – Anaerobic – Facultative • Flocculation: – Happy well fed microbes stick together and settle out taking nutrients colloids etc with them. They become sludge (Bio Solids).

ATP • ATP is Adenosine Triphosphate • High energy phosphate bonded to a five carbon sugar. • Source of all energy for cell activity • Primary reason for the importance of phosphorous as a growth limiting factor in wastewater and environmental systems. (Algae blooms)

Photosynthesis • • • Light Phase Dark Phase Light Phase: Light strikes water (H 2 O Water splits into H 2 and O 2 and an electron. The O 2 is given off and Hydrogen is transferred to a hydrogen acceptor TNP or Triphosphopyridine nucleotide.

Light Phase of Photosynthesis continued • The electron is used by the mitochondria in the cell to convert ADP into ATPand energy is stored in the high energy phosphate chemical bond. • This ATP is used in the Dark Phase f Photosynthesis.

Photosynthesis Dark Phase • ATP + CO 2 +H 20 = PGAL • PGAL is Phosphoglyceraldehyde a three carbon sugar. • 2 PGAL combine to form one six carbon sugar called Glucose. • Glucose is stored in the cell leucoplast for future respiration. • Water is produced as the final Hydrogen acceptor.

Respiration, The flip side of Photosynthesis • • • Respiration is the reverse of photosynthesis Two types of Respiration: Aerobic Respiration. Free atmospheric O 2 Anaerobic Respiration. Chemically bound O 2 In aerobic Respiration Glucose is chemically burned with oxygen in the cell to produce water+ carbon dioxide + Energy.

Photosynthesis vs Respiration

Anaerobic Respiration • When Free oxygen is not available other hydrogen acceptors are used. CO 2, SO 4, NO 2, NO 3 • In anaerobic respiration the carbon substrate is incompletely oxidized producing unstable metabolites. • In Plants and Yeasts : Alcohol and CO 2 produced when glucose is metabolized. (Beer and Bread). • In Animals Glucose is converted to lactic acid in muscles as a temporary hydrogen acceptor.

Digesters are Anaerobic processes • • • Two Stages of Digesters: Acid Stage: Fast. Gas Stage: Slow and sensitive to temperature. Acid Stage: Acid forming bacteria convert food substrate into odorous volatile acids including: Acetic Caproic Valeric Butyric Formic

Gas Stage of anaerobic Digestion. • Gas forming bacteria convert volatile acids into water and gases. • The gases include: – Carbon Dioxide CO 2 35% Plus or Minus – Ch 4, Methane: 70 Percent + or – Hydrogen Sulfide: – Argon:

Bacteria Vital Statistics • • • Rigid unicellular microbes lacking chlorophyll Size 2 X 0. 7 Micron. ; Present everywhere. Mostly Non-Pathogenic. Necessary for higher forms of life to exist because they recycle nutrients.

Methods of identifying Bacteria • • • Multiple Fermentation Tube. Membrane Filter Enzyme Substrate. Gram Staining Specific media.

Bacteria Characterized • Gross Appearance Growth and Morphology including spore forming non-spore forming. • Morphology is shape: • Rod Shaped is called Bacillus : Coli, Typhoid. • Sphere Shaped is called Coccus: Streptococci. • Spirelus is corkscrew shaped: Cholera

Bacteria Characterized • Gross Appearance: Spore former, Non-spore Former Spore a walled resting stage harder to kill. • Morphology- Form: Rod, Sphere, Spiral. • Temperature Range: Cold, Medium or hot loving. • Oxygen Requirement: Aerobic, anaerobic, Facultative.

Bacterial Morphology

Temperature Characterization Psychrophiles or cryophiles Cold Loving Mesophiles: Middle Temp Range. Thermophiles: High Temperature Range. Cryophyllic : Range 2 to 30 C. optimum 12 to 18 C degrees. • Mesophyllic : Range 20 to 45 C. Optimum 25 to 40 C degrees • Thermophyllic: Range 45 to 75 C. Optimum 55 to 65 C degrees. • •

Oxygen Requirement • Aerobic requires free oxygen • Anaerobic can use chemically bound oxygen from: SO 4, NO 2, NO 3, and metal oxides • Facultative can use both metabolic pathways. • When free oxygen is available they usually prefer this because it yields more energy with less work.

Effects of Microbiology on Human Life Expectancy and Mortality Deaths/100, 000 1920 1960 1995 Pneumonia & influenza 207 37 28 Tuberculosis 113 6 <1 Syphilis 16 2 <1 Diphtheria 15 <1 Whooping cough 12 <1 Measles 9 <1 Other (nonmicrobial) 925 909 790 Life expectancy (years) 54 70 76

Increase in Life expectancy is mainly due to: • Between 1920 and 1991, there were 1, 087 deaths associated with waterborne diseases. Of those deaths, 83 percent occurred before 1936 and less than one percent occurred after 1970 Because: • - Sewage treatment and disposal • -Sanitary food handling • -Drinking water treatment • -Development of vaccines, antibiotics and antiseptics

Importance of Wastewater Treatment • Worldwide approximately 1, 2 billion people do not have access to clean and safe drinking water, and 2, to 4 billion people lack sanitation. • Worldwide, the lack of sanitary waste disposal and of clean water for drinking, cooking, and washing is to blame for over 12 million deaths a year. • Every year, 5 million people die of waterborne diseases. • An estimated 4 billion cases of diarrheal disease occur every year, causing 3 million to 4 million deaths, mostly among children.

Water Borne Diseases Waterborne diseases are those acquired through the ingestion of contaminated water. About 80 percent of all diseases are water-related. In many of these illnesses, water infiltrated with sewage spread the disease. An infected person or animal may pass pathogenic bacteria, viruses, or protozoa through waste into the water supply

Collection System Overflows are a serious source of contamination

United States Waterborne diseases 1971 to 1992

New Waterborne Diseases • In the last twenty years a number of pathogenic diseases have appeared. For example: in 1993 in Milwaukee, USA, 400. 000 people fell ill from using drinking water that was contaminated by Cryptosporidium cysts. • In the year 2000, 2. 300 people fell in Walkerton, Canada, because of E. coli O 157: H 7. • In the future more pathogenic microorganisms will emerge and spread through water, because of agricultural magnification, increased population growth, increased migration and climate change. Pathogenic microorganisms can also emerge because they built up resistance to disinfectants and antibiotics.

Pathogenic Bacteria -Typhoid • Typhoid Salmonella typhosa. Exclusively in mans intestine lives 5 months in ice and weeks in soil and feces. In the USA, there approximately 40, 000 cases of Salmonella infection reported each year. [4] According to the World Health Organization, over 16 million people worldwide are infected with typhoid fever each year, with 500, 000 to 600, 000 fatal cases.

Typhoid • Typhoid fever is transmitted by food and water contaminated by the feces and urine of carriers. Polluted water is the most common source. Typhoid fever is characterized by the sudden onset of sustained fever, severe headache, nausea, severe loss of appetite, constipation or diarrhea, mental dullness and meningitis. Most cases in developed countries are imported from endemic countries.

Pathogenic Bacteria Cholera Vibro comma. Endemic in Asia and common in South America. Can be killed with chlorine and 1 week of detention. Using contaminated sewage for fertilizer can result in epidemics of cholera. These diseases can become chronic where clean water supplies are lacking. In the early 1990 s, raw sewage used to fertilize vegetable fields caused outbreaks of cholera in Chile and Peru. The epidemic spread to 19 Latin American countries and has been only partially abated. More than a million cases and 12, 000 deaths have been reported.

Cholera • In 1832, 1. 75% of the New York population was killed as a result of the cholera epidemic in a period of six weeks. This is the equivalent of 140, 000 New Yorkers today. The World Health Organization (WHO) estimates that more than 25, 000 people throughout the world die every day from waterborne diseases. Every year, nearly 1. 5 billion people (mostly children under five) suffer from preventable waterborne diseases such as cholera.

Cholera Symptoms of Choler include high fever and a rice-like diarrhea in which the "rice" is actually pieces of the victim's colon flaking away. It has a short incubation period, from less than one day to five days, and can quickly lead to severe dehydration and death if treatment is not promptly given.

Cholera a Spirochete

Pathogenic Bacteria Gastroenteritis • In 1965 Bacterial Gastroenteritis caused 18, ooo people in Riverside to become sick due to sewerage cross connection, caused by: • Shigella dysenteria • Paratyphoid. • IN 1966 , 18, 000 people in Riverside CA. developed enteritis due to seer contaminated drinking H 2 O

Pathogenic Bacteria Salmonella • Salmonella typhi. Salmonella can survive for weeks outside a living body. They have been found in dried excrement after more than 2. 5 years. Salmonella are not destroyed by freezing. Ultraviolet radiation and heat accelerate disinfection; they perish after being heated to 55 °C (131 °F) for one hour, or to 60 °C (140 °F) for half an hour. To protect against Salmonella infection, it is recommended that food be heated for at least ten minutes at 75 °C (167 °F) so that the center of the food reaches thermal death at this temperature.

Salmonella Outbreaks. • A salmonellosis outbreak began in the spring of 2008 resulting in the illness of hundreds of people throughout the United States. Contaminated fresh peppers tomatoes and cilantro grown on certain farms in Mexico are believed to be responsible. The contamination source is likely to have been wastewater used for irrigation.

Pathogenic Bacteria Leptospirosa Weils Disease, Leptospira is carried in urine by rats can be fatal, attacks kidneys and liver, 100 cases a year. Sewer workers more likely to be aflicted. Dogs and other animals vulnerable. Sea Lions are often affected resulting in periodic die offs.

Pathogenic Bacteria Tetanus • Tetanus (Lock Jaw) Clostridium tetani is a gram positive anaerobic spore forming soil and feces saprophytic none parasitic free living organism that enters cuts and is 19 percent fatal.

Pathogenic Bacteria Tuberculosis • Tuberculosis is Mycobacterium tuberculosis: It can be contacted by aspiration of sewage in to lung and by aerosols. Only case attributed to Wastewater inhalation from worker who fell into activated sludge tank and aspirated sludge.

• Iron Bacteria cause fouling plugging pipes staining clothes. They obtain their energy from the oxidation of ferrous Iron • Crenotrix • Thiobacillus • Ferrrooxidans • Sulfur Bacteria feed on sulfer containing compounds: • Beggiotoa and Thiobacillus • Desulfovibro reduce sulfate to hydrogen sulfide which later may oxidize to produce sulfuric acid which corrodes concrete and metal pipe. H 2 S is very toxic and explosive.

Crown Corrosion Sulfur Desulfovibro Bacteria

Cyanobacteria • Once referred to as Blue Green Algae. Simple none motile chlorophyll bearing bacteria. Cause odor dense matts and some are poisonous. Milk and animals. • Fix nitrogen from the atmosphere. • Some are toxic. Annie, Fannie, Mike. • Anabaena: They are one of four genera of cyanobacteria that produce neurotoxins. • Microcistis • Apanizomenon

Microcystis • Microcystis, a genus of Cyanobacteria is a microscopic organism that is found naturally at low concentrations in freshwater systems such as lakes and streams. • Although not an algae it can be controlled with the application of Aqua Marine Shadow, a blue dye because it photosynthesizes.

Microcystis occasionally forms a harmful algal bloom (HAB). Microcystins are hepatotoxins (toxins that acts upon the liver) and known tumor promoters. If people drink water contaminated by microcystins, symptoms of exposure include nausea, vomiting and, in very rare but severe cases, acute liver failure.

Anabaena

Physical factors affecting Bacteria • Temperature: Enzymes are destroyed at Thermal Death 75 C. Sterilization requires 15 minutes 15 pounds steam, at 121 C. Drying causes spore formation so wet heat is superior to dry heat. • Bacteriostatic agents prevent reproduction. • UV light 2900 Angstrom ionization creates free radicals that destroy DNA. • p. H 6. 8 to 7. 4 is optimum range. Very few bacteria survive below p. H 4 or Above 10 • Electrolytic balance Pickling kills by dehydration, disrupts Osmotic balance.

Factors affecting Bacteria continued • Availability of food c, N, S, P, O 2, H, trace elements. • Presence of oxygen or equivalent hydrogen acceptor.

Growth phases of Bacteria in Batch

Coliform Bacteria Defined • Coliform bacteria are found in the gut of all warm blooded animals. They are not necessarily pathogenic, usually symbiotic, essential for our metabolism, their presence is positive evidence of fecal contamination therefore the coliform test. • Coliform Bacteria are all Aerobic and facultative Anaerobic Gram Negative, Non-spore forming Rod shaped Bacteria that Ferment Lactose Broth in 48 hours at 35 degrees C and produce gas. Remember this definition.

Wastewater Pathogenic Viruses • Sewage treatment may reduce the number of viruses 10 -1000 -fold. • Treatment will not eliminate all virus. • Sewerage Sludge will often contain large numbers of Virus. • Receiving water carries viruses downstream where they remain detectable for a varying period of time, depending on temperature, their absorption onto sediments, and the depth of sunlight penetration. • Enteric viruses can be found in sewage polluted water at the intakes to downstream water-treatment plants.

What are Virus Smallest life form ? Both DNA and RNA types with a Protein coat. DNA and protein coat can be separated and recombined. All are obligate parasites. Not visible with light microscope. There is estimated to be 1 pathogenic virus per million coliform bacteria in sewerage. Coliform test is important surrogate test for enteric pathogenic virus. Turbidity is important because colloidal particles are similar in size to virus.

Norovirus (Norwalk agent) • Norovirus is an RNA virus of the caliciviridae. • Norwalk causes approximately 90% of epidemic nonbacterial outbreaks of gastroenteritis around the world, and may be responsible for 50% of all foodborne gastroenteritis in the US. • Norovirus is transmitted by faecally contaminated food or water, by person-to-person contact, and via aerosolization of the virus and subsequent contamination of surfaces. • After infection, immunity to norovirus is usually incomplete and temporary. There is an inherited predisposition to infection. Individuals with type o blood are more often infected, while blood types B and AB can confer partial protection against symptomatic infection.

Norovirus Symptoms • When a person becomes infected with norovirus, the virus begins to multiply within the small intestine. • After approximately 1 to 2 days, norovirus symptoms can appear. • The principal symptom is acute gastroenteritis that develops between 24 and 48 hours after exposure, and lasts for 24– 60 hours. The disease is usually selflimiting, and characterized by nausea, vomiting, diarrhea, abdominal pain; and, loss of taste. General lethargy, weakness, muscle aches, headache, and lowgrade fever may occur.

Norwalk Virus • Severe illness is rare. • The number of deaths from norovirus in the US is estimated to be around 300 each year, with most of these occurring in the very young, elderly and persons with weakened immune systems. • Symptoms may become life-threatening in these groups if dehydration is ignored or not treated.

Yellow Fever • Yellow Fever Yellow fever and Dengue Fever (Bone Crush) are both viral diseases carried by mosquitoes that bread in contaminated water including septic tanks. Yellow Fever has caused large epidemics in Africa and the Americas. The infection causes a wide spectrum of disease, from mild symptoms to severe illness and death. The "yellow" in the name is explained by the jaundice that affects some patients. The number of people infected with Yellow Fever over the last two decades has increased and yellow fever is now a serious public health issue. There are 200, 000 estimated cases of yellow fever and 30, 000 deaths per year.

Hepatitis Virus Numerous strains A, B, C, D, E. Hepatitis A (formerly known as infectious hepatitis is an acute infectious disease of the liver caused by the hepatitis A virus (HAV), which is transmitted person-toperson by ingestion of contaminated food or water or through direct contact with an infectious person. Tens of millions of individuals worldwide are estimated to become infected with HAV each year. The time between infection and the appearance of the symptoms (the incubation period is between two and six weeks and the average incubation period is 28 days.

Hepatitis A The Hepatitis A virus is viable outside of a host for three months or more. It is killed by exposure to UV light hence detention time in reservoirs exposed to sunlight and aeration is effective. Symptoms: Nausea, Fever, fatigue, Jaundice. Acute liver failure from Hepatitis A is rare. (overall casefatality rate: 0. 5% HAV is excreted in the feces towards the end of the incubation period.

Poliomyelitis Virus • Poliovirus, the causative agent of poliomyelitis is a human entero-cyto-pathogenic virus. • Transmitted by oral fecal route – Therefrore wastwater. • Paralytic disease occurs when the virus enters the CNS and replicates in motor neurons. • Poliovirus is structurally similar to other human enteroviruses such as coxsakievirus and Echovirus. • Wild polioviruses can be found in approximately 10 countries: India, Pakistan, Afghanistan, Sudan, Nigeria, and Egypt, with a 2004 outbreak in West and Central Africa. Wild poliovirus type 2 has probably been eradicated.

Influenza Virus- the Flu • Influenza is an infectious disease caused by an RNA family of viruses, the Orthomysoviridae that affects Birds and mammals. • It is believed to be of avian origin. • Symptoms include chills, fever, soar throat, muscle pain, weakness, nausea, vomiting. • transmitted through the air by coughs or sneezes, creating aerosols or by direct contact with bird droppings, or contact with contaminated surfaces. • Influenza viruses can be inactivated by sunlight or soap.

Influenza • Influenza spreads around the world in seasonal epidemics resulting in the deaths of between 250, 000 and 500, 000 people every year and millions in some pandemic years. On average 41, 400 people died each year in the United States between 1979 and 2001 from influenza. • Three influenza pandemics occurred in the 20 th century and killed tens of millions of people. • Often, these new strains appear when an existing flu virus spreads to humans from other animal species or when an existing human strain picks up new genes from a virus that usually infects birds or pigs from integrated Agricultural practices. An avian strain named H 5 N 1 raised the concern of a new influenza pandemic after it emerged in Asia in the 1990 s

Adenovirus • Adenoviruses are acute respiratory disease causing virus which are transmitted by direct contact, fecal-oral transmission, and occasionally waterborne transmission. • Can establish persistent asymptomatic infections in tonsils, adenoids, and intestines of infected host. Can cause necrotizing pneumonia. • Shedding can occur for months or years and is endemic in parts of the world. • Epidemic conjunctivitis are associated with waterborne transmission, centering around inadequately chlorinated swimming pools and small lakes.

Coxsackie Virus • Coxsackie is an Enteroviruses transmitted by the fecal-oral route. It shares many characteristics with poliovirus. Thus wastewater is a carrier. • Can cause spastic paralysis, infect the heart, pleura, pancreas, and liver, causing pleurodynia, myocarditis, pericarditis, liver. Coxsackie B infection of the heart can lead to pericardial effusion. The development of insulin-dependent diabetes (IDDM) has recently been associated with coxsackievirus B pancreatitis.

E. C. H. O. Virus • ECHO (Enteric Cytopathic Human Orphan) virus, is a type of RNA virus found in the gastrointestinal tract. Exposure to ECHO often causes other opportunistic infections and diseases. Oral Fecal transmission by contaminated water, food, and fomites (inanimate objects) Wastewater. • After infection virus spreads to lymph nodes or is swallowed and reaches the lower gut spreading to heart spleen, nervous system, lungs, bone marrow, liver. • ECHO is the most common cause of aseptic menengitis. Infection of an infant with this virus following birth is associated with high infant mortality rates.

Coliphage • MS 2 Coliphage virus attacks coliform bacteria and is used to test virus removal in disinfection processes because it is a safe surrogate for Enteric pathogenic virus such as polio virus in surface water.

Protozoans • A number of protozoans are pathogens. These including a number of: • Amoeba: Amoebiasis (Entamoeba histolytica) • Ciliates: (Giardia), (Baltidium Coli) • Flagelates: (Cryptosporidium)

Protozoans • Protozoans are single celled or colonial heterotrophs. • One of their important functions in Wastewater processes is their grazing on bacteria like lions control deer. • Identifying protozoans is extremely useful in wastewater process control.

Amoeba Entamoeba Histolitica causes Amoebic Dysenteria. Removal with sand filter and 10 ppm chlorine. Forms cysts found in sludge heavier than water. Move by distorting cytoplasm and cell wall. Flagelates: Zoo and phytoflagellates. Examples Eugena which photosynthesize, zooflagelates move with flagella.

Protozoans Ciliates: move by way of cilia which are organelles on their cell wall. Example of a free swimming ciliate is the Paramecium Example of the stalked ciliate is the vorticella. Example of a pathogen is Baltidium Coli and Giardia.

Sequence of Growth

Protozoan Sequence of Growth

Protozoan forms

Stalked Ciliate

Protozoan Diseases • the 1993 cryptosporidiosis outbreak in Milwaukee that was linked to 104 deaths and illnesses among an estimated 403, 000 people. Cryptosporidiosis is caused by the Cryptosporidium parasite, which is found in water contaminated by human or animal feces.

Giardia • Giardia lamblia is a flagellated Protozoan parasite that colonizes and reproduces in the small intestine, causing giardiasis. The giardia parasite attaches to the epithelium reproducing via binary fission. The Giardia trophozoites absorb their nutrients from the lumen of the small intestine, and are anaerobes.

Giardia • Giardia infects humans, but is also one of the most common parasites infecting cats, dogs and birds. Mammalian hosts also include cows, beaver, dear, and sheep. The main pathways of human infection include ingestion of untreated sewage, and contamination of natural waters also occurs in watersheds where intensive cattle grazing occurs.

Giardia • Giardia infection can occur through ingestion of dormant cysts in contaminated water, food, or by the faecal-oral route (through poor hygiene practices). • The Giardia cyst can survive for weeks to months in cold water, and can be present in contaminated wells and water systems, especially stagnant water sources such as naturally occurring ponds, storm water storage systems, and even clean-looking mountain streams. • They may also occur in city reservoirs and survive water treatment, because Giardia cysts are resistant to conventional water treatment methods such as chlorination and ozonation.

Giardia lamblia

Giardia Life Cycle

Cryptosporidium parvum The main pathways of human infection include ingestion of untreated sewage. C. parvum is considered to be the most important waterborne pathogen in developed countries. It is resistant to all practical levels of chlorination, surviving for 24 hrs at 1000 mg/L free chlorine. Filtration is necessary. The contamination of natural waters also occurs in watersheds where intensive cattle grazing occurs. Cryptosporidium is shed in large numbers by young dairy calves which almost always have the disease. Their manure washes off of fields and lots thus entering the watershed.

• Cryptosporidium parvum is one of several protozal species of cryptosporidium that cause cryptosporidiosis, a parasitic disease of the mammalian intestinal tract. • Primary symptoms of C. parvum infection are acute, watery, and non-bloody diarrhoea. • C. parvum infection is ofserious concern in immune compromised patients, where diarrhea can reach 10– 15 L per day. Other symptoms may include: anorexia, nausia, vomiting and abdominal pain.

C. Parvum • Infection is caused by ingestion of sporulated oocysts transmitted by the fecal-oral route. In healthy human hosts, the median infective dose is 132 oocysts. • Cryptosporidium typically invades the apical tip of the ileal. • Infection causes dehydration and other gastrointestinal symptoms which are generally self-limiting in a healthy host. • Cryptosporidium can be fatal in immune incompetent individuals including those with AIDS or those undergoing immunosuppressive therapy where infection may leading to severe dehydration and, in severe cases, death.

Cryptosporidium parvum

Balantidium coli (Balantidiasis) • Balantidiasis is caused by a ciliate. • Most Balantidial infections are asymptomatic or characterized by intermittent diarrhea or constipation. • Severe infections may cause abdominal pain, colonic tenderness, fever, anorexia, and severe diarrhea. headache, insomnia, nausea, vomiting, pallor, weakness, anemia, dehydration, malaise, and distention. • Stool specimens may be watery and contain blood, mucus, and pus. • Balantidia invade the bowel wall and cause ulcers that resemble amebic ulcers. Immunocompromised complications can be life threatening.

Entamoeba histolytica • There are several different species of amoebae, but the most dangerous, such as Entamoeba histolytica, live predominantly in tropical areas. • These species are able to burrow through the intestinal wall and spread through the bloodstream to infect other organs, such as the liver, lungs and brain. • Amoebic dysentery (amoebiasis) is an infection of the intestine (gut) caused by an amoeba called Entamoeba histolytica that, among other things, can cause severe diarrhoea with blood.

Rotifers are multicellular animals that graze on debris and protozoans. They have a forked tail and are much larger than protozoans.

Rotifer

Rotifer

Parasitic worms • Parasitic worms or helminths are a division of eukaryotic parasites. They live in the digestive tract of and feed off their living hosts, receiving nourishment and protection while disrupting their hosts‘ nutrient absorption, causing weakness and disease. They can live inside humans as well as other animals. Approximately 3 billion people globally are infected with helminths.

Intestinal Helminths • Intestinal helminths represent one of the most prevalent forms of parasitic disease. It is estimated that over a quarter of the world’s population is infected with an intestinal worm of some sort, with round worm, hookworm and whipworm infecting 1. 47 billion people, 1. 05 billion people, and 1. 30 billion people, respectively. Furthermore, the World Bank estimates that 100 million people may experience stunting or wasting as a result of infection. • It is estimated that 400 million, 170 million, and 300 million children are infected with roundworm, hookworm, and whipworm. [ Children may also be particularly susceptible to the adverse effects of helminth infections due to their incomplete physical development and their greater immunological vulnerability.

The Helminth Family • Parasitic worms are categorized into three groups: • Cestodes (tapeworms). • Nematodes (roundworms). Like ascaris • Trematodes (flukes).

Cestodes (tapeworms). • Example is the pork tape worm Taenia solium

Tape worm

Life Cycle of Tape Worm

Beef Tapeworm

Trematodes • Liver Fluke, Shistosomiasis, swimmers itch. • Parasites of mollusks and vertebrates. • Tissue Flukes of different varieties infect bile ducts, lungs, liver (Fasciola hepatica) (Snail host). • Blood flukes inhabit the blood in some stage of their life cycle include species of the genus Schistosoma. (Snail host). • Human infections are most common in Asia Africa, South America, or the Middle East. They can be found anywhere that human waste is used as fertilizer.

Nematode Round Worm • Ascarias, dracunculiasis, elephantiasis, pinworm, hookworm, onchocerciasis, trichinosis, trichuriasis (Whip Worm) • While the natural movement of worms and their attachment to the intestine may be generally uncomfortable for their hosts the migration of Ascaris larvae through the respiratory passageways can also lead to temporary asthma and other respiratory symptoms.

Nematodes The nematodes or roundworms are the most diverse group of all animals over 28, 000 have been described, [of which over 16, 000 are parasitic It has been estimated that the total number of nematode species might be approximately 1, 000.

Fungi • Fungi are small non-chlorophyll bearing plant like organisms. They are powerful decomposers and recyclers of carbon and nutrients. • Fungi share more in common with animals than plants. • Yeasts are important in brewing and bread making. • Filamentous fungi in wastewater play an important roll and can be a problem for settling.

Fungi in composting Fungi are powerful decomposers and break down complex compounds like lignin phenolic compounds, detergents, oil, fat and cellulose. They possess powerful enzymes like protease, lipase cellulase, lipase, and peroxidases. Fungi are found as part of trickling filter slimes on fixed film surfaces where they break down recalcitrant compounds.

Fungi Aspergillus Aspregillus fumagatus and nigra are black molds that are found on windows and wet surfaces. They are often found in association with handling dry sludge or hay. They present a breathing hazard. 1985 CWPCA journal pg. 986 stated that they may cause illness such as brown lung. A problem In inclosed areas where sludge is being handled.

Algae • Autotrophic containing chlorophyl allowing for photosynthesis. • Algae blooms stimulated by phosphorous in excess of 0. 17 mg/l in lakes or. 2 mg/l in streams. Phosphorous is the most limiting factor for growth in fresh water while nitrogen is in salt water environments. Other limiting factors are light, iron, copper, nitrogen.

Algae Cycling cause Fish Kills. • When algae blooms it cycles between photosynthesis in the day and respiration at night. This cycling raises daytime oxygen levels to super-saturation. Because water can only hold 8. 6 mg/l O 2 the rest goes into the air. CO 2 is consumed driving p. H up to as high as 10. At night conditions reverse and respiration depletes all oxygen and saturates water with carbon dioxide driving p. H down to below 5.

Algae Types • Note Blue Green Algae is a Cyanobacteria. Motile Green Algae is a flagellate. • Real Algae Include: • Green Algae like: Isochlorella and Spirogyra. • Brown Algae make up the Diatoms. A characteristic feature of diatom cells is that they are encased within a unique cell wall made of silica (hydrated silicon dioxide) called a frustule. Used as diatomaceous earth in water filters.

Diatoms

Algae Control • All algae are readily controlled by the elimination of light and nutrients. One effective method is a blue dye called Aqua Marine Shadow. Acid Blue 9 is also known as erioglaucine CAS 3844 -45 -9. • Aqua Marine Shadow is available from • F. E. T. Bioremediation Group • 36 Cliffwood Pl. • Santa Rosa, CA 95409 • U. S. A. • 707. 537 -9886 • 925. 457 -4269 cell

Nitrogen Cycle

Nitrogen Cycle

Ponds