low dose organisms
if an organism has a low infective dose, the water treatment process would have to kill off or filter out every last organism to prevent potential illness
resistant organisms
organisms that are resistant to the methods of water treatment, like disinfection
Indicator Organisms and Why do we use them?
Testing for the organisms rather than testing for the contamination
We rely on them for public health as we can not afford and nor would we have the time and resources to test all water for all concerns. So we rely on indicator organisms to indicate that there is a problem. Most of the concerning organisms are associated with feces → so we don’t want to drink water that has feces in it
What are the characteristics of an ideal indicator?
(enteric = intestinal)
Suitable for analysis of all types of water
Present whenever enteric pathogens are present
Survives longer than the hardiest enteric pathogen
Most of the indicator organisms we use DO NOT fulfill this requirement
Does not reproduce in contaminated water
Its level in water reflects the degree of fecal pollution
Detected by highly specific test
Tests are easy to do and sensitive
Harmless to humans
Examples of indicator organisms
Which ones would we use and why
Coliforms
Tells us that there is LIKELY fecal contamination, but it doesn’t say there is ABSOLUTELY fecal contamination
Like E. coli ← the most abundant bacteria in feces
Enterococci
Increasingly used to test brackish and marine water
Studies found a higher correlation to intestinal illness
Also used with refrigerated and frozen foods
More cold tolerant than coliforms
i.e Enterococcus faecalis ← the most common GPC in the intestines
Fecal organisms like nice warm, high nutrient conditions → put them in the cold, low nutrient waters, what would their survival look like?
Studies show that Enterococci in recreational waters have a higher correlation with actual illness
Fecal coliforms
A subset of coliforms
More specific to really identify that these coliforms REALLY ARE of fecal origin
Likely to get FALSE NEGATIVES
Coliphage
Viruses that are specific to E. coli
Even after E. coli dies off, coliphage will still be there
Coliphage stands a better chance of lasting longer and could be more successful as an indicator that could last as long as the hardiest enteric pathogen
Water testing - What are the advantages and disadvantages of each method
No acceptable level for fecal coliforms, enterococci, viruses, or pathogenic protozoans in drinking water
MPN
aka Most Probable Number
Presumptive, confirmatory, and completed tests (photo)
ADVANTAGES
Reliable, easy, cheap, test
Lactose broth is a very nurturing media, so even stressed bacteria may be nurtured into a more viable state
DISADVANTAGES
You only get a statistical estimation
Time-consuming (takes 6 days)
does not differentiate between coliforms and fecal coliforms
Membrane Filtration
Not a required test, but frequently performed b/c it is a quick way of identifying coliform
After filtration, the filter is placed on selective and differential media, incubated, then colonies are presumptively identified and counted
ADVANTAGES
Quicker than MPN
Can filter large quantities of water
You will get an actual count
DISADVANTAGES
You are taking potentially injured/fragile/VBNC organisms, and putting them onto a harsh medium which leads to FALSE NEGATIVES → need to do more tests
HPC
Heterotrophic Plate Count
Shows us an estimate of how many (heterotrophic) bacteria might be in the water
We would not expect there to be a lot of organisms growing in water - if there are, that would indicate a general concern as to why
shows the effectiveness of disinfectants used in water treatment
ADVANTAGES
Shows a range of bacteria, not just potential pathogens
DISADVANTAGES
Doesn’t indicate the specific bacteria (is not specific for coliforms)
Be able to describe 1 method of controlling bacterial growth in food besides using temperature (cooking and refrigerator/freezing)
Modified Atmospheric Packaging (MAP)
Food packed in low O2 or high CO2 limits growth and prevents abiotic oxidation
By modifying the atmosphere of the package, it inhibits microbial growth and prevents fungal growth
Effective and significantly helps to preserve foods w/o damaging those foods or creating any unhealthy byproducts
Food Irradiation
Food is exposed to ionizing radiation, which sterilizes food and improves shelf life
Hazards to workers involved in the irradiation process
Radiation affects food chemistry
Despite being approved by the FDA (as it could possibly save lives if it were utilized), many experts report changes in high protein foods (i.e. rubbery textures, unappetizing odors, etc.)
Bacteriophages
Use of bacteriophages that kill: Listeria, monocytogenes, Salmonella (Salmonelex), E. coli 0157:H7, Campylobacter
Has been approved by the FDA but has not been actively utilized either
Define, in detail, Foodborne Infection versus Foodborne Intoxication
Foodborne Infection
Ingesting of whole microbes that target the intestine
Salmonella, Shigella, Campylobacter, Listeria (very important b/c it has the highest fatality rate of any of these, especially to pregnant women and their fetuses), E. coli (specifically the STECs/0157:07)
A significant cause of morbidity and mortality in the U.S.
Foodborne Intoxication
Results from ingesting exotoxins secreted from bacterial cells growing in food
Caused by organisms such as Staphylococcus aureus or Clostridium botulinum
Know the differences between infection and intoxication
Infection
The infection comes from ingesting the bacteria and then having them grow and cause symptoms in the body
Yes, they produce enzymes and toxins to cause symptoms and damage to the body but, unlike intoxication, it is the growth of the bacteria causing the symptoms
Treating with an antibiotic could be successful
Longer incubation period
More likely to have fevers and lower intestinal symptoms
Intoxication
Intoxication comes from ingesting toxins in the food, those toxins got there by bacteria growing in the food, but it is not the bacteria that cause the symptoms, it is the toxins ALREADY produced.
Again, it is purely the toxin causing the illness
Treating with an antibiotic would be worthless
Have to recover on your own or be treated with an antidepressant
Onset times are quicker
More likely to have vomiting
Discuss why it is important to know and understand the difference between an infection and intoxication (think of 3 important points related to diagnosis, treatment, and prevention)
Food infection and food intoxication are 2 types of food poisoning. However, both of these terms are in contrast to the diagnosis, treatment, and prevention. Knowing the differences between food infections and food intoxication is essential to ensure effective management and recovery.
Food infection is caused by pathogens when ingested. Treat with an antibiotic? Prevent food infections by heating your food properly and avoiding raw milk and unpasteurized dairy products and juices.
On the other hand, food intoxication is primarily caused due to the presence of toxins in the food consumed. Vomiting is a key symptom of intoxication because the body wants to immediately get rid of the toxin and the fastest way to do that is by vomiting it out. A prolonged bout of vomiting can cause the body to be dehydrated so drink plenty of liquids. Prevent food intoxication by cleaning your food, your kitchen area, and your hands before and when handling food; as well as properly chilling food and cooking food.
Examples of each (infection and intoxication) (names of bacteria and characteristics of those various ones we learned about before - Staphylococcus aureus, Bacillus cereus, Clostridium perfringens, Clostridium botulinum, Salmonella, E. coli 0157:H7, Campylobacter, Listeria)
Staphylococcus aureus
Food intoxication
Causes Staphylococcal enteritis
Very common; symptoms come on rapidly; usually nonfatal
Bacillus cereus
Food intoxication
Causes Bacillus cereus enteritis
Mimics staphylococcal enteritis but is less acute; usually self-limited (ultimately resolving itself w/o treatment)
Clostridium perfringens
Food infection
Causes Perfringens enterotoxemia
Vegetative cells produce toxins within the intestine
Clostridium botulinum
Food intoxication
Causes Botulism (note: toxin is sensitive to heat)
Rarely seen in commercially prepared foods; can be fatal
Salmonella
Food infections (usually caused by S. typhimurium and S. enteritidis)
Causes Salmonellosis
Very common; can be severe and life-threatening
E. coli 0157:H7
Food infection
Causes Escherichia enteritis
The cause of hemolytic uremic syndrome (a condition that affects blood and blood vessels)
Campylobacter
Food infection (usually caused by C. jejuni)
Causes Campylobacterosis
Very common; animals are carriers of other species; low infective dose
Listeria
Food infection
Causes Listeriosis
Most severe in fetuses, newborns, and the immunodeficient
Concerns in milk - raw milk and pasteurized milk (think about the article too)
Milk comes from a living source and contains many bacteria naturally
Difficult to control bacteria growing in and on cows
Contamination during the milking process
10 coliforms/ml is the standard for raw milk
No matter how clean a dairy may be, it is difficult to be sure that cows are not carrying any potential pathogens, even frequent testing will not detect all, that is why consuming raw milk IS a risk
Pasteurized milk
Pasteurization reduces the number of bacteria and should kill off pathogens
Phosphatase test
Checks for the presence of phosphatase, the enzyme is destroyed by heat and should not be present after pasteurization
Microscopic counts for bacteria and white blood cells
High numbers of white blood cells could indicate an ill cow in the mix
Antibiotic residual
It is crucial that a cow that has been on an antibiotic does not contribute milk for a specified amount of time so there is no chance that residual antibiotics can still be in the system
There are many individuals who have antibiotic allergies
Food, Water and Milk Discussion:
BE PREPARED TO HAVE ONE OF THESE QUESTIONS ON YOUR EXAM 4
what are ARGs and, based on this article, why are they a concern
ARGs are antimicrobial resistance genes and they are a concern because they are transferrable and can thus pass to other microorganisms leading to more antibiotic resistance in the environment.
what is AMR and, based on this article, why is it a concern
AMR is antibiotic resistance and they are a concern because if more bacteria are resistant to our antibiotics, we are less able to treat infections and illnesses; what was once an effective treatment can be void.
based on this article, why would there be a concern about leaving milk at room temperature
Leaving milk at room temperature is a concern because it would enrich the ARGs; the milk would become a reservoir of ARGs.
based on this article, what does this mean for the US and low-income countries
According to the article, one recent study evaluated the risk factors associated with the carriage of antibiotic-resistant E. coli from people in northern Tanzania, and direct microbial transmission in raw milk was found to be the primary predictor of the prevalence of AMR. This demonstrates that raw milk would maintain reservoirs of ARGs and transmit antibiotic resistance.
based on this article, what are the different types of bacteria found and are they considered to be pathogens
Many different types of bacteria were found but especially Pseudomonadaceae; it was dominant in raw, vat-pasteurized, and ultra-pasteurized milk, while Streptococcaceae was more prevalent in high-temperature short-time pasteurized milk samples. Both are considered to be pathogens.
what types of conclusions (if any) can be drawn from this study
Pasteurization matters. Raw milk has a distinct microbiota with a higher abundance of viable bacteria containing antimicrobial resistance genes, both of which are amplified by spontaneous fermentation at room temperature. This work indicates that raw milk consumption poses an additional risk to consumers through the transfer of antimicrobial resistance genes.
Microbial Death definition
Permanent loss of reproductive capability even under ideal conditions
In other words... The irreversible loss of the ability to grow
Hard to detect, microbes often reveal no conspicuous vital signs to begin with
How microbes die off – Pattern of microbial death
Microorganism populations are not killed instantly but die off according to first-order kinetics - which determines that it is a linear die-off (Chick’s Law)
Meaning… the cells in a population die at a constant rate
Population death usually occurs exponentially
Even VBNC cannot become viable again
D value
Decimal reduction time
The time required to kill 90% of cells
So, it’s a measure of an agent’s killing efficiency
Must be sure persister cells (VBNC condition) are dead
Once they recover they may regain the ability to reproduce and cause infection
Concept and importance of VBNC microbes
Viable But Not Culturable
Retains virulence
A threat to public health and food safety due to their non-detectability through conventional food and water testing methods
Are there any factors that impact the ability of these methods to work properly? (ex. extraneous organic materials, nature of the microbes you are trying to control, amounts of contaminants, environment)
Only when enough molecules of an essential protein or gene are damaged will the cell die. Several factors influence the speed at which lethal damage accumulates:
Initial population size (the larger the population, the longer it takes to decrease it to a specific number)
Population composition (are spores involved?)
Agent concentration, or dose of radiation
Duration of exposure
Presence of organic material (such as blood or feces) that can inhibit disinfectant action; also called “organic load”
Modes/Mechanisms of Action of Antimicrobial Agents - you should have had an opportunity to look at these in our Disinfectants, Antiseptics, and Use Dilution Lab AND in the Norton SmartWorks Required Assignment - which ones to know for the exam will be discussed in the lecture - I didn't ask you to memorize any of these mechanisms)
Sections 13.2 and 13.3 in the book
Moist and dry heat
Moist heat kills better than dry heat, thanks to the ability of water to penetrate cells.
Filtration
For heat-sensitive media or chemical additives
A form of sterilization
Filtration through a filter with a pore size…
0.2 um → removes bacteria and bacterial endospores, yeasts, mold spores, algae, and protozoa
20 nm → removes viruses
Irradiation
Can kill pathogens in foods w.o damaging the food itself
food is bombarded with high-energy electromagnetic radiation
UV does not penetrate liquids or solids well; it is useful only for surface sterilization
Gamma rays (cobalt-60), electron beams, and X-rays are more effective at irradiation
A potent strategy for sterilizing food after harvesting
A microorganism’s sensitivity to irradiation depends on a lot of factors
Irradiation of food is effective in eliminating parasites and bacteria but is inadequate for eliminating viruses or prions
Halogens – Chlorine, Iodine
Disinfectant
Corrodes tissues; destroy proteins
Used for: disinfection of drinking water, pools, and hospital equipment; sanitation; wound antisepsis
Gas highly toxic
Phenolics
Disinfectant
Denatures proteins
Standard use
Can cause dermatitis; vapor affects the lungs
Alcohols
Disinfectant
Cause dehydration; denatures proteins; dissolves membrane lipids
Used for: cleaning of instruments; cleaning of the skin
Dries skin; toxic if ingested
Hydrogen Peroxide
Disinfectant
Hydroxyl radicals attack membrane lipids, DNA, protein
Used for: wound cleaning; sterilization of contact lenses
Will whiten skin
Aldehydes
Glutaraldehyde
Disinfectant
Alkylation damages protein, DNA, and RNA
Used for: disinfection of medical equipment
Irritates skin, mucous membranes
Formaldehyde
Disinfectant
Reacts with protein, DNA, and RNA; cross-links proteins and DNA
Used for: embalming; preservation of anatomical specimens
Pungent odor, irritating fumes; carcinogen
Heavy Metals
Bisphenols?? Is BPA a heavy metal? meh
Disinfectant
Disrupts membranes and proteins
Used for: disinfection of solid surfaces
Very toxic if swallowed
Quaternary ammonium compounds (cationic agents)
Disinfectant
Disrupts membrane
Used for: sanitation of floors, walls, blood pressure cuffs
Toxic if ingested; irritates mucous membranes
Therapeutic Index
The ratio of the toxic dose to therapeutic dose
Toxic dose: maximum dose tolerated by the patient
Therapeutic dose: minimum dose per kg of body weight that stops growth
Selective toxicity
Antibiotics kill or inhibit bacteria w/o damaging the eukaryotic organisms
Modes/Mechanisms of Action of Chemotherapeutic Agents/Antibiotics (5) and 1 example of each
(photo)
Antibiotic Resistance Mechanisms - describe 1 (needs to be at least a few sentences) (How the bacterium is resistant to the antibiotic)
Ribosome protection (or rescue): Gram-positive organisms can produce proteins (for example, MsrE or TetO) that bind to ribosomes and dislodge antibiotics bound near the peptidyltransferase site.
Several of the important essay questions are going to be related to:
Indicator organisms - what they are, why they are important, what makes an ideal one, examples of those currently used, important advantages and disadvantages
Foodborne illnesses - what is the difference between FB Infection versus FB Intoxication, why would it be important to know, what are examples of each
The diversity found in bacteria -what examples of bacteria or groups of bacteria represent the diverse world of bacteria