Lab Manual

Experiment Name: Treatment of Wastewater by UV Disinfection

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1. Introduction

As the number of people and creation increases, they need accumulated water use, making a
corresponding rise in waste product amount. This accumulated water use and method waste
product generation needs additional economical elimination of by-products and pollutants that
enables for effluent discharge among established environmental regulative limits. Waste
product treatment consists of many steps, the last of that is medical aid. Correct waste
product medical aid will forestall diseases like infectious hepatitis, Cholera, and stomach flu
that are caused by the microorganism, protozoan cysts and viruses. The
foremost wide used methodology of medical aid is halogen disinfection. Nonetheless, Ultraviolet
(UV) remedial aid is turning into additional well-liked and has several advantages.

UV disinfection works by shifting the magnetic force energy from a mercury arc
light to AN organism’s polymer and polymer, efficiently terminating the organism’s ability to
breed. a number of the benefits to ultraviolet medical care embrace, the inactivation of most pores,
viruses, and cysts; no ought to handle poisonous and/or risky chemicals. As actinic ray medical
care could be a physical method, thus it's simple to implement and needs shorter contact time, and
no harmful residual effects for human or aquatic life. Once it compared to Cl medical care, actinic
ray medical care is costlier. Loads of care is taken to make sure that the amount of muddiness and
total suspended solids are low enough so that they won't interfere with the actinic ray medical
care method.

Coliform microorganism, underneath the family of the enterobacteriaceae, reside within

the viscus tract of humans and are excreted in massive numbers in faces of humans
and homothermic animals. The term “total coliform” in laboratory testing refers to all or
any coliform microorganism from feces, soil or alternative origins. Untreated domestic sewer
water usually contains over three million coliforms per a hundred milliliter. E. coli (E.
coli) microorganism are a set of coliform bacteria and are a lot of related to unwellness.
However, just one strain (or species) of E. coli is really morbific (disease-causing) and is
thought as E. coli

1.1 Objectives

Objectives of this experiment are

 To check the effectiveness of wastewater UV disinfection to remove Enterobacteriaceae and
E. coli and
 To function in an environmental laboratory and analyze the samples using dilution methods
and without cross-contamination.

2 Technical Theory

Some microbes got to be controlled so as for North American country to take care of our
current customary of living. Pathogens and microorganisms that pamper food are
of explicit concern as are microbes that fade nonliving matters. One in every of the tools we have
a tendency to use for dominant. The unfold of microbes are antimicrobial agents. Disinfectants are
antimicrobial mediators that make up two categories: microstatic disinfects, that inhibit the
expansion and replica of microbes and microcidal disinfectants that destroy the microbes. Specific
microbes respond otherwise to disinfectants and this suggests that no single agent is effective
against all microorganisms. Many factors influence however a disinfectant works. These
factors embrace make-up of disinfectant, time of contact with microbes, pH, sensitivity of
microbes, and also the presence of extraneous material. There are three classes of
disinfectant based mostly upon however they have an effect on the microbes. The
primary interferes with cell walls (phenols, alcohols, detergents), the second is interference
with protein operate (detergents, significant metals), and also the third
is macromolecule denaturation (phenols, alcohols, detergents). Data of however a disinfectant
works will build it easier to decide on one for every specific purposes.

The disinfection of treated waste via ultraviolet (UV) radiation could be a physical method that in
the main involves passing a movie of wastewater inside shut proximity of a UV supply (lamp).
The potency of UV medical care depends on the physical and chemical water quality
characteristics of the waste before disinfection. With a {more robust an improved} quality
of waste comes a more economical UV disinfection method. UV disinfection doesn't end in an
enduring residual within the waste. this can be a drawback once waste should be piped
or keep over vital distances and time (particularly relevant to use schemes) as re-growth of
the microbe population is taken into account a risk The log reduction achieved by
a decontamination process is a measure of how thoroughly the process reduces
the concentration of a contaminant. It is defined as the common logarithm of the ratio of the levels
of contamination before and after the process, so an increment of 1 corresponds to a reduction in
concentration by a factor of 10. The equations used in this experiment are given below

𝐴
Log Reduction = 𝑙𝑜𝑔10 [1]
𝐵

Where,

A= concentration of a contaminant before the treatment

B= concentration of a contaminant after the treatment

𝐴−𝐵
Percent Reduction = × 100 [2]
𝐴

Where, A and B are the same components as explained in the earlier equation

P = (1 − 10−𝐿 ) × 100 [3]

Where,

P = Percentage Reduction

3 Materials and Equipment

The materials and equipment used in this experiment were Sterile water dilution bottles (99 mL of
sterile water), (6) E. coli/Coliform count plates , (6) Enterobacteriaceae count plates, 3M Petrifilm
Spreader, Incubator , Rubbing alcohol, 1 mL sterile pipets, Permanent marker, Safety Glasses and
Latex gloves. Cell cultures and micro-organisms were incubated for 24 hours in a controlled
atmosphere. In the standard incubator, the temperature is controlled.

4 Safety Guidelines

 Always wear latex gloves and safety glasses when in contact with the wastewater.
 Use hand sanitizer or wash hands immediately after removing the gloves.
 Do not touch your face after contact with wastewater; wash your hands thoroughly before
touching your face.
 If you are wearing gloves and you touch the wastewater samples or count plates, your gloves
become contaminated. While wearing contaminated gloves, do not touch anything that you or
someone else might come in contact with later
5 Experimental Procedure

The 1:100 dilution of pre-clarifier sample was prepared by using a sterile 1 mL pipet and then
added 1 mL of the wastewater in the dilution bottle. Closed the cap and labeled the bottle with the
group number and sample name. Then the 1:100 dilution of the post-clarifier/pre-disinfection
sample was prepared by using the same method used above but new sterile pipette was used to
avoid the contamination.

Three E. coli/Coliform Count Plates and three Enterobacteriaceae Count Plates were selected and
placed them on a level surface and labeled the each count plate. The 1 mL of the sample was placed
evenly over the count plate to inoculate each count plate. Completely filled the circle to avoid
from the sample to squeeze off the plate. Lifted the top film of the count plate and emptied the 1
mL sample from the pipette onto the center of the count plate. Then gently rolled down the top
film to spread the sample and prevented the trapping of air bubbles and incubated the plates at a
temperature of 35oC for 24 hours.

After 24 hours, plates were removed from the incubator and counted the bacteria colonies and
counted each plate for the three times to avoid from the more variance in the result.

5.1 Washing the hands after collecting and/or handling wastewater samples

After the gathering or handling of wastewater samples hands are washed to get rid of soil, dirt, and
microorganisms. If water and soap isn't out there, hands are often clean with ash instead.
Hand laundry with soap systematically at important moments throughout the day prevents
the unfold of diseases like symptom and infectious disease that are transmitted through fecal-oral
routes. Folks will become infected with metabolic process diseases like grippe or the respiratory
disease, for instance, if they are doing not wash their hands before touching their eyes, nose, or
mouth.
One of the disadvantages of V.V disinfection is that it does not provide the residual disinfecting dose.
Moreover, certain organisms develop the ability to repair the damaging effect in the near V.V or violet
spectral range by photoreactivation