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Biological Treatment of Hospital Wastewater by using Batch and Rotating

Biological Contractors

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 International Journal of Advanced Science and Technology
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Biological Treatment of Hospital Wastewater by using Batch and Rotating
Biological Contractors
Krishna Prasad. M1, Jyothi.K2, Swetha Sri. V3& Nagaraju. G4
2
Department of Botany, Andhra University, Visakhapatnam
1,3,4
Department of Chemical EngineeringGMR Institute of Technology- Rajam, Srikakulam, A.P.
2
jyothikaparapu@gmail.com

Abstract
The wastewater generated from the hospital consists of pharmaceuticals, pathogenic
bacteria, drugs, suspended particles and bloodstains that may influence the ecosystem
possibly causes cancers that have been observed for decades. To overcome these, impacts,
the wastewater is started to treat before discharged to the water bodies. The treatment
process has evolved into various physical, chemical and biological methods. Biological
treatment methods are conducted in this paper to study the comparative analysis for the
BOD and COD removal efficiencies in the batch study and RBC model. A pilot scale of a
biofilm model of Rotatory Biological Contactor (RBC) was implemented as the extension of
batch experiments. The prepared synthetic hospital wastewater with high concentrations are
treated using the bacteria Pseudomonas aeruginosa and Vibrio cholera as a treatment
media. The substrate removal rate was analyzed by conducting the BOD and COD
experiments in the batch study. The Pseudomonas aeruginosa results indicated 78% BOD
removal efficiency from the batch study. This bacteria was used in the treatment of influent
by using an attached biofilm model RBC of 6 liters capacity with 4 discs, each having
surface area 0.4m2 and 45% submergence. Compared with batch experiments, RBC has
greatly removes the 67% of COD and 85% of BOD. This work also illuminates the
importance of Pseudomonas aeruginosa bacteria for the effective elimination proficiency of
a substrate in the cure of pharmaceutical discharge. The treated effluent will be reused for
decreasing the increasing demand for water. In the present study the biofilms have maximum
of dry matter and are at ease to dehydrate and settle in contrast to activated sludge. RBC
can significantly diminish the treatment price, related to commercial incineration processes
or advanced oxidation processes,. This work proves RBC is operative for an innately bio
obstinate wastewater even at lesser ambient temperature.
Keywords: Batch experiments, Rotatory Biological Contactor (RBC), Pseudomonas aeruginosa,
Vibrio cholera, COD and BOD

1 Introduction:

Hospitals are generally consuming an extensive amount of water ranging between 400 to 1200 liters/day
and equally creates the same volume of wastewater load which generates from different segments
including patient wards, clinical wards, surgery units, ICU, laboratories and holds a relatively inconstant
composition liable on the activities consists (Kumarathilaka et al., 2016). The effluent wastewater
comprises pharmaceuticals that are partially metabolized, pathogenic microorganisms, toxic chemical
substances, drugs, and radioactive elements. The effluent release criteria for hospital wastewater should
regard to EPA 1986 and the tolerance limits in India is according to BIS standards IS 4764 Hospitals are
generally consuming an extensive amount of water.

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There is an increase in production and consumption of pharmaceuticals considerably during the last
decades, their yearly manufacture beyond thousands of tons (Santos et al., 2007; Sim et al., 2011). The
omnipresence of pharmaceuticals in surface water, groundwater and drinking water, with absorptions
wavering from ng L-1 to µg L-1 has been reported by many authors (de Jesus Gaffney et al., 2016; Koreje
et al., 2016). The occurrence of these compounds of the ecosystem will provoke the pollution by
involving biological imbalance and causes endocrine disruptions, bacterial resistance, ecotoxic effects
leads to death of marine biota (Pauwels and Verstraete, 2006; Chen et al.,2003de Jesus Gaffney et al.,
2016; Zhao et al., 2016; Kostich et al., 2014; Yan et al., 201). Antibiotics in wastewater may encourage
bacterial opposition, even at less contents (Hernandez et al., 2007). The by-products of hospital discarded
fluid are high-strength biological wastewater that is obstinate to biological treatment processes. In
general, hospital wastewaters (HWWs) are straightly quitted, lacking any treatment, into the gutter system
to be co-cured with municipal wastewaters (Al Qarni et al., (2016)). Generally, it is cured by incineration
in the pharmaceutical plant, however, this choice is appropriate for big initiatives because of its massive
investment and large operating cost (Tchobanoglous et al ., 2003; Yang et al., 2009). Inopportunely,
conventional Waste Water Treatment Plants are generally planned to isolated suspended solids and to
remove decomposable organic matter and they are commonly unable to entirely reduce pharmaceuticals,
their abolition reliant on their adsorption on biotic sludge and biodegradation (Santos et al., (2007).

To eradicate such risks, it is crucial to treat the influent wastewater and yield effluent free from pollutants
(Langenhoff et al.,2013). To remove such pollutants chemical treatment processes such as osmosis,
advanced oxidation processes, ozonation, photocatalysis, activated carbon, ultrasound oxidation, etc.,
which are expensive and energy consumptive. In contrast, biological treatment techniques are robust and
cost-effective when compared to chemical treatment (Langenhoff et al., 2013). It includes activated
sludge process, trickling filter, membrane bioreactor, up-flow anaerobic sludge blanket, aerated ponds,
stabilization lagoons, fluidized bed reactor, Rotatory Biological Contactors, and others wherein the
microbial groups are used as a treatment media for the deprivation of pollutants (Stalin, 2014). The
biological process is frequently used because it yields an elimination competence changing among 51 %
to 80% at a hydraulic retention time of 1- 4.5 days (Jeswani&Mukherji, 2015). Therefore, the fate of the
pollutants should be considered and a suitable treatment technique should be adopted.

The rotating biological contactor (RBC) biofilm (attached-growth) process was promoted in the 1970s
(Najafpour et al.,2005). Current curing amenities for municipal and industrial wastewater management
with intermediate strength (COD concentration is 600–2000mgL−1) of biological substances requires
RBC as an essential biofilm process (Akunna and Jefferies, 2000; Vidal et al., 2002). Biodegradation of
organic substances within the RBC depends on the biomass composition of the discs (Torkian et al.,2003;
Zheng et al.,2010). The advantages of RBC are the preservation of high concentration of microorganisms
in the exponential state, more capable for degrading biological composites, more aptness for fluxes in
movement rate and biological concentration (Chen et al.,2006; Wang et al.,2009), nitrification and
denitrification along with removal carbon (Hiras et al.,2004; Tawfik et al.,2005), easy operation, less
water in the sludge forming it to settle, less energy intake (Torkian et al.,2003), and requirement of small
land area (Taseli et al., 2008). Thus, an economically feasible process for managing real high strength
organic pharmaceutical wastewater is required.

In this research, the potential of the biological process is investigated to treat the prepared synthetic
pharmaceutical wastewater. The prepared synthetic wastewater was treated by conducting batch studies
and fabricated Rotatory Biological Contactor (RBC) on a small scale. The microorganisms Pseudomonas
aeruginosa and Vibrio cholera are used as a treatment media to treat the wastewater. The dry weight
density of microorganisms was also calculated to define the microorganisms count. The cultured
microorganisms are used in the batch study and the microorganism which gives the best removal
efficiency was used in the RBC process as a biofilm. This work aimed to determine the treatment result
computed by evaluating the removal of Chemical Oxygen Demand (COD) and a 5-day test Biological

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Oxygen Demand (BOD5) in batch study and RBC. Investigation of the discharges COD and BOD needs
to be done and the values obtained used to establish an empirical relation of COD to BOD5. An
evaluation of BOD with COD measures whether the compound is readily decomposable. For BOD5, a
sign is that a COD: BOD proportion of more than 100 means that the compound is comparatively non-
biodegradable and a ratio of less than 10 is somewhat degradable (Prashant et al.,2013)

2 Materials and methods

2.1 Synthetic wastewater preparation

Synthetic hospital Waste Water was arranged in agreement with Seyhi et al., (2013). The synthetic
wastewater was prepared with different 9 assortments of tablets of every assortment of 2 tablets and by
adding glucose, (NH4)2SO4, KH2PO4, NH4Cl, and CaCl2 are diluted each 1mg/l in distilled water. For
maintaining microbial movement Nitrogen and phosphorus were added in excess to the synthetic effluent.
The concentrations of nitrogen, phosphorus, and glucose, are adapted to sustain a C: N: P ratio of
100:10:5. The concentrations of pharmaceuticals were selected according to those which detected in real
hospital effluents (Santos et al., 2013; Yilmaz et al., 2017). The prepared stock solution is kept for a
contact time of 96 hours and put in storage in dark at temperature 4 oC. The COD and BOD tests are
conducted for this prepared synthetic wastewater (Ouarda et al.,2018).

2.2 Analytical methods

Chemical and biochemical oxygen demand (COD & BOD) were the main factors selected to investigate
the RBC treatment process. The COD and BOD of prepared wastewater samples were estimated earlier
and later treatment. Analyses of COD, BOD5, were conducted to study the biodegradability of the
pharmaceutical discharges. The BOD is the amount of dissolved oxygen needed by microbes for the
oxidation of living content in wastewater in aerobic situations. The BOD5 test is a bioassay method
relating to the extent of oxygen spent by the bacteria while steadying the living material in the waste in
laboratory settings. The test is engaged for 5 days at 20°C (68°F) (APHA, 1989). COD tests were done
according to the Standard Methods (APHA 1992) by the ferrous ammonium sulfate method (APHA,
1989). Dissolved Oxygen (DO) was conducted by Winkler’s method (APHA, 1998). The pH value of
water was measured by pH meter (Systronics, India). Turbidity was determined by Nephelo - turbidity
meter. Additionally, microorganisms were observed by a microscope.

2.3 Microorganism’s characteristics:

Biological treatment involves microbial consortia as a treatment media.In the present work, the biological
treatment of the prepared wastewater involved two microorganisms Pseudomonas aeruginosa and Vibrio
cholera.

Table.1. Characteristics of microorganisms

Characteristics Pseudomonas aeruginosa Vibrio cholera

Gram type Negative Negative
Domain Bacteria Bacteria

Family Pseudomonadaceae Vibrionaceae

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Shape Rod shape Coma shaped

Size 1-5µm long, 0.5-1.0 µm wide 0.3µ diameter, 1.3µ in length

Aerobe or Aerobic (at times facultative Facultative anaerobe

anaerobe anaerobic)
Colour Blue-green, red-brown Yellow-green
Diseases cause respiratory tract problems, skin cholera, diarrhea and vomiting,
problems, Ear and Eye coma and death.
infections, bones and joints
problems and blood and heart
diseases

2.4 Dry weight density of microorganisms:

From the subculture stock, a 10ml sample was taken and centrifuged at 6000 rpm for 20 minutes. After
centrifugation is completed, testers from the supernatant were studied for biomass composition which
were detected by filtering from 0.45 milli pore filter papers and defining dry weight after dehydrating in
an oven until stable weight (Kargi, F. 1996).

Dry weight density= (initial weight-final weight)*1000/ (volume of sample taken)

2.5 Experimental Setup

2.5.1 Batch experiment

In the batch mode, the experiments are carried out at an ambient temperature of 250C. The batch
experiments are carried out in the various flasks of 300ml capacity. Three 300ml flasks packed with 10ml
of Pseudomonas aeruginosa and wastewater and other three 300ml flasks are packed with 10ml of Vibrio
cholera and wastewater. These flasks were used to calculate the removal efficiency of COD and BOD.
The flasks were allowed in aerobic conditions for an incubation period of 5 days (Salama et al., 2015).

2.5.2 RBC Setup

In this study, the performance of laboratory Rotatory Biological Contactor (RBC) was tested for the
treatment of prepared synthetic wastewater. The RBC unit is fabricated with a semi-circulated trough of
volume 6litres capacity and provided with a PVC shaft on which discs are attached with a width of
200mm and the shaft is provided with the support of two bearings at each end. The rotating disks provide
the development of microbes of the sewage, which break down and steady organic pollutants. The shaft is
fixed overhead the water level in the tank. A motor of 30 RPM is used in this experiment. The motor can
vary its RPM from 1 to 30. Oxygen is acquired from the atmosphere as the discs revolve. As the microbes
progress, they construct upon the media until they shred off owing to shear forces by the revolving discs
in the wastewater. Effluent from the RBC is then passed through final clarifiers where the micro-
organisms in suspension settle as sludge. The sludge is withdrawn from the clarifier for further treatment
(Boyles, W. 1997; Donald et al., 2001). The experiment work is carried out in aerobic conditions. This
RBC work is intended to study the effective removal of COD and BOD with the help of Pseudomonas
aeruginosa (Wilson & Lee1997).

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Table:2. Physical Parameters of Device

Physical parameter Value

Rotational speed (rpm) 2

Tank capacity (lts) 6

Number of discs 4

Diameter of disc (mm) 200

Surface area of the disc (mm2) 9.4x103

Diameter of discs (mm) 5

Space between the discs (mm) 100

Submergence (%) 45%

3 Results and Discussions

3.1 Dry weight density calculation of biomass

Biomass dry weight denotes the volume of the mass of microorganisms expressed as g/ml of sample. The
volume of biomass is calculated by a count of the number of cells per one ml of solution. The removal
efficiency of treatment depends on the number of microorganisms in the aeration tank. The presence of
more number of cells results in higher efficiency [Kargi, 1996].

Dry weight density= [(initial weight-final weight) / (volume of sample taken)] *1000

The dry weight density of Pseudomonas aeruginosa is 9180 mg/l and Vibrio cholera is 5130 mg/l.

3.2.Treatment of synthetic effluent in batch studies

The treatment performance of the batch study was evaluated for BOD analysis. The synthetic influent was
characterized by a COD content of 2000mg/l and a BOD range of 8600mg/l. The batch experiment is
conducted based on the concentration of initial COD concentration and the dilutions are prepared at
concentrations of 2000mg/l, 1500mg/l, and 1000mg/l. Two microorganisms are used separately in each
set of flasks to evaluate the removal efficiency capacity of those microorganisms. The batch mode results
shown that the Pseudomonas aeruginosa gives the best removal efficiency (Fig.1).

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3.2.1 BOD removal efficiency by Pseudomonas aeruginosa

Fig:1. Comparison of BOD values between standard effluent and Pseudomonas aeruginosa

The graph gives the details of BOD removal efficiency in prepared synthetic wastewater before and after
conducting a batch study experiment by adding Pseudomonas aeruginosa as a treatment media after an
incubation period of 5 days. Three concentrations of 8600mg/l, 6450mg/l, and 4300mg/l are the initial
BOD values before conducting a batch study. The BOD values without microorganism after incubation
period are decreased to 7500mg/l from 8600mg/l, 2000mg/l from 6450mg/l, and 4000mg/l from 4300mg/l
due to the natural degradation of organic matter. At the same time, BOD values with organism after
incubation period are highly degraded from 7500mg/l to 3200mg/l, 6450mg/l to 600mg/l and 4300mg/l to
1100mg/l.

3.3.Treatment of synthetic effluent in Rotatory Biological Contactor (RBC):

RBC experimentwas conducted as the extension of the batch experiment. The evaluated samples from the
batch study are treated in the RBC to determine the higher efficiency. Pseudomonas aeruginosa results in
the higher efficiency in the batch study. Hence this microorganism is used in the RBC as a treatment
media.

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3.3.1 COD removal efficiency of RBC by Pseudomonas aeruginosa

Fig:2. COD removal by Pseudomonas aeruginosa of standard effluent and RBC effluent

The COD removal efficiency by Pseudomonas aeruginosa of prepared synthetic wastewater is shown in
fig.2. The initial concentration of COD is 900mg/l before conducting the RBC experiment. The
wastewater without organism reduces COD by itself from 900mg/l to 620mg/l. After the treatment with
the organism in the RBC for 5 days is reduced to 300mg/l.

The quantity and kind of bacteria of the attached biofilms of the RBC starts adapting to new wastewater
or reproduced deviations fit for new wastewater. (Chen et al.,2003; Eckenfelder,2000)

3.3.2 BOD removal efficiency in RBC by Pseudomonas aeruginosa

Fig:3. BOD removal efficiency in RBC with Pseudomonas aeruginosa

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Figure 3 represents the changes of BOD5 removal during acclimation presents the BOD removal
efficiency by Pseudomonas aeruginosa of prepared synthetic wastewater. The initial concentration of
BOD is 900mg/l. The wastewater without organism reduces BOD by itself from 900mg/l to 670mg/l.
After the treatment in RBC with organism for 5 days is greatly reduced to 125mg/l.

The BOD5 elimination diminished to around 60% according to the accumulation of pharmaceutical
wastewater enlarged. This is due to the rate of pharmaceutical wastewater addition was collecting rapidly
related to the initial addition rate and the microorganisms are not able to acclimatize in time. When the
additive amount of pharmaceutical wastewater was adjusted and the acclimation period became lengthier,
the removal of BOD5 starts to increase to about 80% in the end. This shows that the microbes in the
biofilms slowly amended to pharmaceutical wastewater and they capably detached the decomposable
organics. The above-mentioned trials indicate, even at less ambient temperature, the RBC system is
operative for treating pharmaceutical wastewater. This indicates that this RBC technology can
theoretically be used in reserved conditions.

3.3.3 COD removal efficiency of wastewater by Pseudomonas aeruginosa

Fig:4. Comparison between COD removal by Batch & RBC system

COD removal efficiencies in Batch experiments result in a slight increment because of the improper
collection of the sample. RBC gives the best removal results of COD, due to the high degradation of
pollutants by Pseudomonas aeruginosa which is provided with support media.

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3.3.4.BOD removal efficiency of wastewater by Pseudomonas aeruginosa

Fig:5. Comparison between BOD removal by Batch & RBC system

The BOD5 removal efficiency of both batch experiments by Pseudomonas aeruginosaresults in the best
removal efficiency of 78% and 86.1%. The graph represents the increment, it is due to the presence of
support media provided for the microbial consortia for their growth. The BOD5elimination efficacy
reduced firstly with growing influent concentration, and then it steadied.

3.4. Treatment of Pharmaceutical Wastewater with RBC.

After acclimation, the outcome of influent COD concentration on the COD, and BOD5 deduction from
pharmaceutical wastewater was studied. After wastewater runs into the RBC device, the organic
complexes are gradually decomposed by microbes. There is greater organic content in wastewater close to
the inlet which causes microbes to replicate quicker. On the contrary, there is comparatively fewer
organic content in the downstream stages causing a lesser amount of microorganisms. Therefore, as the
water continues downstream, the amount of microbes on the stage lessens. Related with the mud in
activated sludge methods (Tchobanoglous et al., 2003) the amount of dry matter in the biofilms is greater
and the biofilms have more density that is easy to settle. In this experiment, RBC technology can
knowingly reduce the treatment cost and organic concentration. The key features influencing the
treatment of RBC comprise the kind of wastewater, influent concentration, stages of RBC, and
temperature.

The air and liquid temperatures flanked by 24-26°C respectively and as a result, the biomass grew on the
RBC disks noticeable within 3 to 4 days of operation which was a thick biofilm layer on each side of a
disk. The biofilm varied from dark to light golden-brown color under aerobic conditions initially. Under
high substrate and good growing conditions, it can be hypothesized that the high reaction rate in the
biomass will cause a high oxygen uptake and a subsequent rapid oxygen depletion in the reactors. The
circulation of oxygen into the biofilm and liquid mass is inadequate and further controlled by the low disk
rotating speed and relatively high temperature. Similar results were noticed by Kim et al. (1985) in
treating wastewater.

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3.5.Concentration Vs Efficiency

80

78

76

74

72

70

68

66
0 500 1000 1500 2000 2500

Fig:6. Removal efficiency

The graph shows the relation between influent concentration and removal efficiency. As the concentration
decreases the efficiency is increases.

3.6.Outcome of Influent Concentration on elimination of COD

When the organic input concentration was below 1000mgL−1, the COD removal efficacy is near to 50%.
With an increase of organic input concentration from 400 to 800mgL−1, the COD elimination improved
somewhat. The total COD removal was not more related to the outcome of treating sewage wastewater.
This is due to the existence of biorecalcitrant substances in pharmaceutical wastewater (Yang et al.,
2009). The COD elimination proficiencies of varied kinds of wastewater progress consequently with
rising ambient temperature, (Badawy et al.,2009; Gebhardt and Schroder, 2007).

3.7.Outcome of Influent Concentration on elimination of BOD5

The BOD5values of the inlet and outlet water were measured to study the biodegradability of
pharmaceutical wastewater. Initially, with increasing influent concentration, the BOD5removal efficiency
decreased. This indicates that microorganisms decompose most of the biodegradable content of
pharmaceutical wastewater. The microbes in biofilms unable to destroy all of the organics in wastewater
because of the existence of some toxic substances biorecalcitrant and the low ambient temperature. This
has an effect on the BOD5 exclusion to some extent.

The present results indicated that a rise in the organic filling rates resulted in only a modest decline in the
elimination efficiency, while the deterioration in biomass extended progressively. Even when organic
overloading had developed throughout, efficiencies of 90-91 % were still obtained. It has been noticed
that the high bacterial activity supported by the constant high ambient temperature permitting rapid
substrate utilization of the present high strength synthetic wastewater. It is also observed that the system
although operating under organic overloading conditions acts effectively as a facultative process with the
air-exposed biofilm being in an aerobic condition and the submerged biofilm being in an anaerobic phase.

Based on the effects of influent concentration on COD, and BOD5removal, it is concluded that the
influent concentration does not substantially influence organic exclusion. This is due to the existence of
various stages in the RBC. The early phase safeguards the shock of influent concentration and toxins. One

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more cause of resistance to more influent concentration is that the biofilms have a chunkiness of 0.5–3
mm, and the shock of input organics control the interior layer of the biofilms smaller than the exterior
layer, thus leaving a moderately active inner layer. Thus it is challenging to degrade the entire piece of
biofilm from one disc or all the biofilms from all disks. Therefore, RBC is proficient in repelling the
effect of influent concentration variation. This means that RBC has a robust aptness for the concentration
shock of wastewater related to the activated sludge method. After such a shock, it can reestablish its
purpose quickly. Nevertheless, related to activated sludge practice, it is only suitable for wastewater with
low influent concentration (Chen et al., 2003). With the decline of COD and BOD5downstream, metabolic
actions of microbes declined and their oxygen intake reduced. This leaves additional oxygen in the waste.
In extra, the RBCs are efficient at reaerating the water. The ambient temperature has an important
influence on treatment ability with greater temperature causes more effective treatment (Su, 2010).

4 Conclusion

The lab-scale rotating biological contactor (RBC) was operated while treating synthetic wastewater
utilizing Pseudomonasaeruginosa as a microbial consortia used in the RBC. The RBC experimented
aerobically which acts as an efficient method for handling high strength wastewaters under the rotational
speed of discs shaft of 2 rpm and with the submergence of 45%. The microorganism reduces the influent
COD from 1000mg/l to 300mg/l which was a 70% removal efficiency and also reduces the BOD of
influent from 1500mg/l to 125mg/l achieved an efficient removal of 86% at HRT 60h.

5 Acknowledgments:

One of the authors JyothiKaparapu appreciatively admits financial support of UGC, New Delhi (Post-
Doctoral Fellowship for women) No. F.151/2014-15/ PDFWM-2014-15-AND-26595(SAII).

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