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Bioassay Study of Biomedical Liquid Waste (Treated Efﬂuent) of Bhopal City,

Madhya Pradesh, India
Article · January 2020

DOI: 10.4444/ijrr.1002/1649
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International Journal of Research and Review
Vol.7; Issue: 1; January 2020
Website: www.ijrrjournal.com
Original Research Article E-ISSN: 2349-9788; P-ISSN: 2454-2237
Bioassay Study of Biomedical Liquid Waste (Treated

Effluent) of Bhopal City, Madhya Pradesh, India
Reeta Kori, Alok Saxena, Sandhya Mokhle, Vineeta Goutam,
Ankita Kulshreshtha, Vandana Ramnani, Makhan Parmar
Central Laboratory, Madhya Pradesh Pollution Control Board, Paryavaran Parisar, E-5, Arera Colony, Bhopal,
Madhya Pradesh, India
Corresponding Author: Ankita Kulshreshtha
ABSTRACT immunization of human beings or animals

or in research pertaining thereto or in the
Biomedical liquid waste originated from production or testing. [1] Medical care is
hospital, laboratories & research centres has the vital for our life, health and well being but
potential to pose serious threat to the the waste generated from medical activities
environment in terms of spread and
can be hazardous, toxic and even lethal
contagiousness of infectious diseases to the
system. Total fourteen hospitals were selected because of their high potential for diseases
for this study. The study was conducted during transmission. [2] Hospital liquid waste is
April 2018 to March 2019. Monitoring of wastewater generated from all activities of
biomedical liquid waste (Treated Effluent) was the hospital as medical and non medical
performed as per guidelines of Central Pollution activities from the operating, emergency &
Control Board and bioassay test was performed first aid, laboratory, diagnosis, radiology,
as per BIS method, 6582: 2001. Biomedical kitchen and laundry activities. [3-5] Hospital
waste may affected the aquatic life when it liquid waste refer to infectious, non-
discharged in to the surface water. Fish are the hazardous and hazardous liquid waste with
primary bio indicator as they are very sensitive sufficient free liquid arising from dental,
towards the pollution, so this study was based
medical, nursing, pharmaceutical or similar
on toxicity test on fresh water Brachydanio rerio
(Commonly known as Zebra fish). The practice, care, investigation, treatment that
observation based on 90% survival of tested requires additional safety packaging to
fishes at 50% dilution (1:1 ratio of waste water ensure safe transportation, treatment and
to test water by volume) up to 96 hrs, that storage. [6-7] In most developing countries,
represented as toxicity factor one. Further there is lack of technology, skills to
increase in toxicity factor represents increase in implement and monitor hospital waste
toxicity found acute state. This study concluded management programs. [8-10] Hospital
that treated effluents from most of monitored wastewater includes macro- and micro-
hospitals of Bhopal city were found free from pollutants of wide concentration range from
acute lethal toxicity. laboratories, research units, operation
rooms, units, where medicine, nutrition
Key Words: Hospitals, Biomedical Liquid
Waste, Effluent Treatment Plant, Treated solutions are prepared and polyclinics. [11-12]
Effluent, Bioassay Test, Toxicity factor As a result of such consumption, the
wastewater contains significant amounts of
1. INTRODUCTION microorganisms, heavy metals, toxic
Bio-medical waste means any solid chemicals and radioactive elements.
or liquid waste including its container and Hospital originated wastewater is
any intermediate product, which is discharged to city sewage systems in many
generated during the diagnosis, treatment or countries, treated together with domestic
International Journal of Research and Review (ijrrjournal.com) 164

Reeta Kori et.al. Bioassay study of biomedical liquid waste (treated effluent) of Bhopal city, Madhya Pradesh,
India
wastewater and discharged to receiving tissues, microbes etc. It is defined as

environments. [13-14] Hospital laboratory estimation or determination of concentration
wastewater is considered a mixture of or potency of a physical, chemical or
pathogen microorganisms. The genetic biological substance (agent) by means of
structure of such microorganisms may be measuring and comparing the magnitude of
altered by the direct or indirect effect of the response of the test with that of standard
wastewater components and lead to bacteria over a suitable biological system under
with high antibiotic resistance. [15] Many standard set of conditions. [21] This study
developed countries required disinfection of was based on bioassay test for analysis of
hospital wastewater before being discharged toxicity of treated effluent from various
into sewage systems. Currently the most hospitals of Bhopal city.
commonly used disinfectants are liquid
chlorine, NaClO, ClO2, and O3. [16- 17] The 2. METHODOLOGY
fact that hospital wastewater contains entero 2.1. Study Area
bacteria and enteric pathogens, poses a risk Bhopal city is the capital of Madhya
for public health. Today, the marked Pradesh which is very well connected to all
increase in antibiotic resistance of infectious the corners of the country situated in the
agent pathogen bacteria seen both in central part of India. Its latitude is 23°25’99
nosocomial and community-acquired N and longitude is 77°.41’26 E. Bhopal is
infections is one of the most important also known as the “Lake City” for its
problems. [18] As a matter of fact, heavy various natural as well as artificial lakes and
metals pose threat to environment and is one of the greenest cities in India. [22]
human health since they are not biologically
degradable pollutants and that they are 2.2. Monitoring Locations
movable pollutant sources. [19-20] Total fourteen hospitals of Bhopal
Bioassay is an assay designed to city were selected for this study which are
analyze any compound by use of a depicted in table no1 and shown in figure 1.
suitable biological system like animals,
Table 1: Monitoring Locations
S.N. Code Monitoring Locations
1. H1 Bansal Hospital Shahapura, Bhopal
2. H2 Bhopal Fracture Hospital & Surgical Centre, Arera Colony, Bhopal
3. H3 Bhopal Memorial Hospital and Research Centre, Bhopal
4. H4 Chirayu Hospital, Bairagarh Kalan, Bairagarh, Bhopal
5. H5 Gastrocare Liver and Digestive Disease Centre, Arera Colony, Bhopal
6. H6 Government Shakir Ali khan(Gas Rahat) hospital, Bhopal
7. H7 Hamidia Hospital, Royal Market, Bhopal
8. H8 Indira Gandhi Mahila And Child Hospital, Bhopal
9. H9 Jawaharlal Nehru Cancer Hospital Idgah Hills, Bhopal
10. H10 J K Hospital & Medical Research Centre in Kolar Road, Bhopal
11. H11 L B S Hospital, Motia Talab Road Bhopal
12. H12 Kamla Nehru Hospital (Gas Rahat), Bhopal
13. H13 People's Hospital Bhanpur, Bhopal
14. H14 Swami Vivekanand Regional Spine Centre, M P Nagar, Bhopal
2.3 Monitoring and Analysis

Monitoring of treated effluents from effluent treatment plant of various selected hospitals was
performed as per guidelines of Central Pollution Control Board and Bioassay test performed
as per BIS method- 6582: 2001. [23]

India
Figure 1: Monitoring Locations
3. RESULT & DISCUSSION

This study resulted by observations of bioassay test which are depicted in table no 2 to 16. In
table no 2, toxicity factor (Tf) was found one during all four quarters observation of bioassay
test of treated effluent from Bansal Hospital Shahapura, Bhopal.
Table 2: Bioassay test of treated Effluent from Bansal Hospital (H1), Shahapura, Bhopal
Quarter Dilution Ratio of waste Parts of dilution water pH (pH Unit) Dissolve Oxygen %
Factor water to test water to be added to one part (mg/l) Survival
(by volume) of Waste water Before After Before After
(by volume) Test Start 96 hrs Test Start 96 hrs
1 (Apr- 1 1:1 0 6.98 7.02 5.5 3.0 100*
June 2018)
II (July- 1 1:1 0 7.10 7.18 4.5 3.2 100*
Sep 2018)
III (Oct- 1 1:1 0 6.82 6.86 5.2 3.1 100*
Dec 2018)
IV (Jan- 1 1:1 0 6.85 6.92 5.3 2.8 100*
Mar 2019)
Remark : *100% survival seen in observed Toxicity factor = dilution factor ( Tf 1 for 1:1 Ratio of Waste water to test water by volume) after
96 hrs. Standard Tf =1 represent test condition standard dilution water containing 100 mg/L Potassium dichromate, all fish should
survive.Fish Species used for test: Zebra Fish (Brachydanio rerio) Fish size-30±5 mm and wt- 0.2 to 0.3 gm, Temp: 25±1.0oC.
In table no 3, toxicity factor (Tf) was found one during all two quarters observation of
bioassay test of treated effluent from Bhopal Fracture Hospital & Surgical Centre, Arera
Colony, Bhopal.
Table 3: Bioassay test of treated effluent from Bhopal Fracture Hospital & Surgical Centre (H2), Arera Colony, Bhopal
Quarter Dilution Ratio of waste Parts of dilution water pH (pH Unit) Dissolve Oxygen %
1 (Apr- # # # # # # # #
June 2018)
II (July- # # # # # # # #
Sep 2018)
III (Oct- 1 1:1 0 7.25 7.33 5.0 2.9 100*
Dec 2018)
IV (Jan- 1 1:1 0 6.95 7.08 4.8 2.5 100*
Mar 2019)
96 hrs. Standard Tf =1 represent test condition standard dilution water containing 100 mg/L Potassium dichromate, all fish should survive. .
#Monitoring started after second quarter. Fish Species used for test: Zebra Fish (Brachydanio rerio) Fish size-30±5 mm and wt- 0.2 to 0.3
gm, Temp: 25±1.0oC.

India
In table no 4, toxicity factor (Tf) was found one during all four quarters observation of
bioassay test of treated effluent from Bhopal Memorial Hospital and Research Centre,
Bhopal.
Table 4: Bioassay test of treated effluent from Bhopal Memorial Hospital and Research Centre (H3), Bhopal
Quarter Dilution Ratio of Waste Parts of dilution water pH (pH Unit) Dissolve Oxygen %
1 (Apr- 1 1:1 0 7.13 6.99 4.5 3.8 100*
June 2018)
II (July- 1 1:1 0 6.94 8.18 4.8 2.5 100*
Sep 2018)
III (Oct- 1 1:1 0 8.07 8.12 4.5 4.0 100*
Dec 2018)
IV (Jan- 1 1:1 0 6.55 6.81 6.8 5.1 100*
Mar 2019)
In table no 5, toxicity factor (Tf) was found one during all four quarters observation of
bioassay test of treated effluent from Chirayu Hospital, Bairagarh Kalan, Bairagarh, Bhopal.
Table 5: Bioassay test of treated effluent from Chirayu Hospital (H4), Bairagarh Kalan, Bairagarh, Bhopal
1 (Apr- 1 1:1 0 7.68 7.72 5.8 3.1 100*
June 2018)
II (July- 1 1:1 0 7.62 7.75 8.2 6.1 100*
Sep 2018)
III (Oct- 1 1:1 0 7.02 7.64 5.4 3.9 100*
Dec 2018)
IV (Jan- 1 1:1 0 7.11 7.28 6.9 5.3 100*
Mar 2019)
96 hrs. Standard Tf =1 represent test condition standard dilution water containing 100 mg/L Potassium dichromate, all fish should survive.
Fish Species used for test: Zebra Fish (Brachydanio rerio) Fish size-30±5 mm and wt- 0.2 to 0.3 gm, Temp: 25±1.0oC.
In table no 6, toxicity factor (Tf) was found one during all four quarters of bioassay test
observation of treated effluent from Gastrocare Liver and Digestive Disease Centre, Arera
Colony, Bhopal.
Table 6: Bioassay test of treated effluent from Gastrocare Liver and Digestive Disease Centre (H5), Arera Colony, Bhopal
1 (Apr- 1 1:1 0 7.21 7.38 4.1 2.8 100*
June 2018)
II (July- 1 1:1 0 7.16 7.19 4.5 2.9 100*
Sep 2018)
III (Oct- 1 1:1 0 6.73 6.77 4.4 3.8 100*
Dec 2018)
IV (Jan- 1 1:1 0 7.08 7.18 5.8 2.5 100*
Mar 2019)
In table no 7, toxicity factor (Tf) was found two during first quarter and one during all other
quarters of bioassay test observation of treated effluent from Government Shakir Ali khan
(Gas Rahat) hospital, Bhopal.

India
Table 7: Bioassay test of treated effluent from Government Shakir Ali khan (Gas Rahat) hospital (H6), Bhopal
1 (Apr- 2 1:2 1 6.31 6.45 6.9 5.5 100*
June 2018)
II (July- 1 1:1 0 8.23 8.38 7.1 6.3 100*
Sep 2018)
III (Oct- 1 1:1 0 7.91 7.93 6.2 5.0 100*
Dec 2018)
IV (Jan- 1 1:1 0 7.96 7.99 6.5 5.2 100*
Mar 2019)
Remark : *100% survival seen in observed Toxicity factor = dilution factor ( Tf 1 for 1:1 Ratio of Waste water to test water by volume
similarly 2 is for 1:2) after 96 hrs. Standard Tf =1 represent test condition standard dilution water containing 100 mg/L Potassium
dichromate, all fish should survive. Fish Species used for test: Zebra Fish (Brachydanio rerio) Fish size-30±5 mm and wt- 0.2 to 0.3 gm,
Temp: 25±1.0oC.
In table no 8, toxicity factor (Tf) was found two during third quarter and one during all other
quarters of bioassay test observation of treated effluent from Hamidia Hospital, Royal
Market, Bhopal.
Table 8: Bioassay test of treated effluent from Hamidia Hospital (H7), Royal Market, Bhopal
1 (Apr- 1 1:1 0 7.16 7.31 6.3 4.8 100*
June 2018)
II (July- 1 1:1 0 7.66 7.69 5.6 4.3 100*
Sep 2018)
III (Oct- 2 1:2 1 6.56 6.78 6.2 4.1 100*
Dec 2018)
IV (Jan- 1 1:1 0 6.85 6.93 5.8 3.6 100*
Mar 2019)
Temp: 25±1.0oC.
In table no 9, toxicity factor (Tf) was found one during all quarters of bioassay test
observation of treated effluent from Indira Gandhi Mahila and Child Hospital, Bhopal
Table 9: Bioassay test of treated effluent from Indira Gandhi Mahila and Child Hospital (H8), Bhopal
1 (Apr- 1 1:1 0 6.96 7.18 6.6 3.8 100*
June 2018)
II (July- 1 1:1 0 6.18 6.25 6.6 3.5 100*
Sep 2018)
III (Oct- 1 1:1 0 5.61 5.71 7.1 6.8 100*
Dec 2018)
IV (Jan- 1 1:1 0 7.23 7.31 7.7 6.5 100*
Mar 2019)
In table no 10, toxicity factor (Tf) was found two during fourth quarter and one during all
other quarters of bioassay test observation of treated effluent from Jawaharlal Nehru Cancer
Hospital Idgah Hills, Bhopal.

India
Table 10: Bioassay test of treated effluent from Jawaharlal Nehru Cancer Hospital (H9), Idgah Hills, Bhopal
1 (Apr- 1 1:1 0 7.96 8.18 4.5 2.0 100*
June 2018)
II (July- 1 1:1 0 7.73 7.81 5.8 3.2 100*
Sep 2018)
III (Oct- 1 1:1 0 7.36 7.38 4.1 2.7 100*
Dec 2018)
IV (Jan- 2 1:2 1 7.15 7.28 4.5 2.8 100*
Mar 2019)
Temp: 25±1.0oC.
observation of treated effluent from J K Hospital & Medical Research Centre, Kolar Road,
Bhopal.
Table 11: Bioassay test of treated effluent from J K Hospital & Medical Research Centre (H10) Kolar Road, Bhopal
1 (Apr- 1 1:1 0 7.23 7.36 5.8 2.5 100*
June 2018)
II (July- 1 1:1 0 6.99 7.21 6.6 2.8 100*
Sep 2018)
III (Oct- 1 1:1 0 7.11 7.19 5.6 2.3 100*
Dec 2018)
IV (Jan- 1 1:1 0 7.28 7.35 6.5 2.9 100*
Mar 2019)
In table no 12, toxicity factor (Tf) was found four during fourth quarter and one during all
other quarters of bioassay test observation of treated effluent from L B S Hospital, Motia
Talab Road Bhopal.
Table 12: Bioassay test of treated effluent from L B S Hospital (H11), Motia Talab Road Bhopal
1 (Apr- 1 1:1 0 7.09 7.18 6.2 3.8 100*
June 2018)
II (July- 1 1:1 0 7.28 7.33 5.6 2.8 100*
Sep 2018)
III (Oct- 1 1:1 0 7.21 7.26 4.7 2.5 100*
Dec 2018)
IV (Jan- 4 1:4 3 6.51 6.66 5.5 4.5 100*
Mar 2019)
similarly 2 is for 1:2, 4 is for 1:4) after 96 hrs. Standard Tf =1 represent test condition standard dilution water containing 100 mg/L
Potassium dichromate, all fish should survive. Fish Species used for test: Zebra Fish (Brachydanio rerio) Fish size-30±5 mm and wt- 0.2 to
0.3 gm, Temp: 25±1.0oC.
observation of treated effluent from Kamla Nehru Hospital (Gas Rahat), Bhopal.

India
Table 13: Bioassay test of treated effluent from Kamla Nehru Hospital (Gas Rahat) (H12), Bhopal
1 (Apr- # # # # # # # #
June 2018)
II (July- 1 1:1 0 7.46 7.48 5.8 3.2 100*
Sep 2018)
III (Oct- 1 1:1 0 6.85 6.96 7.2 6.1 100*
Dec 2018)
IV (Jan- 1 1:1 0 7.52 7.68 7.7 6.8 100*
Mar 2019)
#Monitoring started after first quarter. Fish Species used for test: Zebra Fish (Brachydanio rerio) Fish size-30±5 mm and wt- 0.2 to 0.3 gm,
Temp: 25±1.0oC.
observation of treated effluent from People's Hospital Bhanpur, Bhopal
Table 14: Bioassay test of treated effluent from People's Hospital (H13), Bhanpur, Bhopal
1 (Apr- 1 1:1 0 7.25 7.28 4.1 2.5 100*
June 2018)
II (July- 1 1:1 0 7.86 7.96 4.8 3.2 100*
Sep 2018)
III (Oct- 1 1:1 0 7.47 7.52 5.2 3.1 100*
Dec 2018)
IV (Jan- 1 1:1 0 6.16 6.25 6.8 5.1 100*
Mar 2019)
In table no 15, toxicity factor (Tf) was found two during fourth quarter and one during all
other quarters of bioassay test observation of treated effluent from Swami Vivekanand
Regional Spine Centre, M P Nagar, Bhopal.
Table 15: Bioassay test of treated effluent from Swami Vivekanand Regional Spine Centre (H14), M P Nagar, Bhopal
1 (Apr- # # # # # # # #
June 2018)
II (July- 1 1:1 0 7.28 7.31 5.1 2.5 100*
Sep 2018)
III (Oct- 1 1:1 0 6.56 6.77 4.2 2.8 100*
Dec 2018)
IV (Jan- 2 1:2 1 7.56 7.76 5.6 3.8 100*
Mar 2019)
similarly 2 is for 1:2, 4 is for 1:4) after 96 hrs. Standard Tf =1 represent test condition standard dilution water containing 100 mg/L
Potassium dichromate, all fish should survive. #Monitoring started after first quarter. Fish Species used for test: Zebra Fish (Brachydanio
rerio) Fish size-30±5 mm and wt- 0.2 to 0.3 gm, Temp: 25±1.0oC.
Summary of Bioassay test observations of studied Hospitals of Bhopal city is depicted in

table no 16.

India
Table 16: Summary of Bioassay test observations of studied Hospitals of Bhopal city.
Code I Quarter II Quarter III Quarter IV Quarter
H1. Toxicity factor (Tf) was Toxicity factor (Tf) was Toxicity factor (Tf) was Toxicity factor (Tf) was found
found one* found one * found one * one *
H3. Toxicity factor (Tf) was Toxicity factor (Tf) was Toxicity factor (Tf)was Toxicity factor (Tf) was found
found two* found one* found one * one *
found one* found one * found Two* one *
found one* found one * found one * two*
H11. Toxicity factor (Tf) was Toxicity factor (Tf) was Toxicity factor (Tf) was The toxicity factor (Tf) was
found one* found one * found one * found Four *
found one* found one * found one * two*
Remark: *100% survival seen in observed Toxicity factor = dilution factor ( Tf 1 for 1:1 Ratio of Waste water to test water by volume
similarly 2 is for 1:2, 4 is for 1:4 ) after 96 hrs. Standard Tf =1 represent test condition standard dilution water containing 100 mg/L
Potassium dichromate, all fish should survive.Fish Species used for test: Zebra Fish (Brachydanio rerio) Fish size-30±5 mm and wt- 0.2 to
0.3 gm, Temp: 25±1.0oC.
The observation based on 90 % Besides the above immediate effects,

survival of tested fishes at 50 % dilution harmful substances are adsorbed through the
(1:1 ratio of waste water to test water by gills, skin or intestine, thereby impairing
volume) up to 96 hrs, that represented as physiological functions. These effects may
toxicity factor one. Further increase in ultimately lead to the death of fish.
toxicity factor represents increase in toxicity
found acute state. The fish may not always 4. CONCLUSION
die but their reproductive capacity, Bio-medical waste is generated during
breeding, spawning and development are the diagnosis, treatment or immunization of
adversely affected. In aquatic toxicology human beings, animals or from research
fish have been widely and popularly activities pertaining there to or in the
acclaimed as a test species for evaluating production or testing of biological fluids. It
the potency of toxicants to cause lethacity include wastes like human anatomical
(acute toxicity) or any other sub lethal waste, animal waste, microbiology &
responses, using selected behavioral biotechnology waste, discarded medicines
biochemical or physiochemical and & cytotoxic drugs, soiled waste, solid waste,
hematological responses. Bio medical waste liquid waste, incineration ash, chemical
effluents are complex and can be polluted wastes etc. These wastes are very hazardous
by non-biodegradable end toxic organic because of the potentially infectious in
compounds and are a serious threat to the nature as it may pose a serious threat to
environment. Fish are affected by toxic human health, if its management is
substances chiefly in following two ways: indiscriminate and unscientific. The study
A- Epithelia -absorb toxic substances, concluded that treated effluents from most
getting damaged in this process, for of monitored hospitals of Bhopal city were
instance, the gills stick together or get found free from acute lethal toxicity
congested with mucus and get destroyed. B-

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Bioassay Study of Biomedical Liquid Waste (Treated Efﬂuent) of Bhopal City,

Article · January 2020

Alok Saxena Ankita Kulshreshtha

30 PUBLICATIONS 222 CITATIONS

The user has requested enhancement of the downloaded file.

Bioassay Study of Biomedical Liquid Waste (Treated

ABSTRACT immunization of human beings or animals

International Journal of Research and Review (ijrrjournal.com) 164

wastewater and discharged to receiving tissues, microbes etc. It is defined as

2.3 Monitoring and Analysis

International Journal of Research and Review (ijrrjournal.com) 165

Figure 1: Monitoring Locations

3. RESULT & DISCUSSION

International Journal of Research and Review (ijrrjournal.com) 166

International Journal of Research and Review (ijrrjournal.com) 167

International Journal of Research and Review (ijrrjournal.com) 168

International Journal of Research and Review (ijrrjournal.com) 169

Summary of Bioassay test observations of studied Hospitals of Bhopal city is depicted in

International Journal of Research and Review (ijrrjournal.com) 170

The observation based on 90 % Besides the above immediate effects,

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20. Ortega L M, Lebrun R, Blais J F and 22. https://en.wikipedia.org/wiki/Bhopal

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