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DESALINATION
S i" '1

ELSEVIER Desalination 152 (2002) 315-324 i

www.elsevier.com/locate/desal

Advanced treatment for municipal wastewater reuse in

agriculture. UV disinfection: parasite removal and by-product
formation

Lorenzo LibertP*, Michele Notarnicola b, Domenico Petruzzelli b

Department of Civil and Environmental Engineering, Polytechnic University of Bari, viale del Turismo 8,
74100 Taranto, ltaly
°Tel. +39 (080) 5963-368; Fax +39 (080) 5963-282; email: l.liberti@poliba.it
bTel. +39 (080) 5963-477; Fax +39 080 5963 635; emails: notarnicola@poliba.it, d.petruzzelli@poliba.it

Received 30 March 2002; accepted 12 April 2002

Abstract

This paper reports the experimental results of a pilot-scale (100 mVh) investigation, carded out at the West Bari
(S. Italy) municipal wastewater treatment plant, focused on parasite removal and disinfection by-product (DBP)
formation during the UV disinfection of clarified (CL) and clarified-filtered (F) secondary municipal effluents at
doses necessary for achieving the Italian microbial limit for unrestricted reuse ofwastewater in agriculture (2 CFU/
100ml of total coliforms). The investigation demonstrated that parasites like Giardia lamblia cysts and
Cryptosporidium parvum oocysts were both significantly affected by UV radiation and that potential UV-promoted
formation of DBPs (nitro-phenols and N-nitroso-amines) did not occur according to GC/MS and LC/MS analytical
evidences. O&M costs ranged from ~ 17.5 up to E 35/1000 m3for effluent F and CL respectively.

Keywords: Cryptosporidium; Disinfection; Disinfection by-products; Giardia; Treatment costs; UV rays; Wastewater
reuse

*Corresponding author.

Presented at the EuroMed 2002 conference on Desalination Strategies in South Mediterranean Countries:
Cooperation between Mediterranean Countries of Europe and the Southern Rim of the Mediterranean.
Sponsored by the European Desalination Society and Alexandria University Desalination Studies and Technology
Center, Sharm El Sheikh, Egypt, May 4-6, 2002.

0011-9164/02/$- See front matter © 2002 Elsevier Science B.V. All rights reserved
PII: SOOl1-9164(02)01079-2
316 L. Liberti et al. /Desalination 152 (2002) 315-324
1. Introduction
Municipal wastewater contains a variety of
pathogenic organisms of human origin. Diseases
caused by these pathogens can occur as a result
of ingestion of untreated or improperly treated
water, ingestion of infected aquatic food species,
skin contact with contaminated water and with
improperly disinfected wastewater effluent in reuse
application. Such diseases are more likely in coun-
tries characterised by scarce rainfall, lack of fresh-
water resources and high groundwater salinity as in
the Mediterranean basin where agricultural reuse
of municipal wastewater is becoming a compulsory
choice for water resources manage-ment.
Various schemes of advanced (or tertiary)
treatment have been proposed in the last two
decades with the so called "full Title 22" scheme
[1 ], i.e. secondary effluent further submitted to
clariflocculation, sand filtration and final disinfec-
tion, most often adopted so far. The increasing
occurrence ofbio-resistant microorganisms and new
pathogenic species, in particular, makes advanced
disinfection a key-step for municipal wastewater
reuse in agriculture [2].
Actually, municipal wastewater effluents are
commonly disinfected by chlorination. However,
protozoa like Cryptosporidium parvum and
Giardia lamblia and helminths like Nematodes,
of particular concern causing lethal diseases in
immunocompromised populations, have demon-
strated to be resistant to chlorine-based disinfection
procedures [3,4]. Furthermore, chlorine is known
to raise serious toxic effects on living organisms.
In fact, it can react with organics contained in
municipal wastewater to form various toxic chlo-
rinated hydrocarbons, such as trihalomethanes and
related disinfection by-products (DBP), known as
animal carcinogen and suspected to be carcino-
genic towards human being [5,6].
Accordingly, attempts are under way world-
wide to address the effectiveness of safer alternative
disinfectants able to meet the stringent microbial
standards usually required for wastewater recla-
mation and reuse in agriculture [7].
In this context, a 3-year R&D project partially
supported by the European Community within the
framework of the Avicenne Iniziative was initiated
in 1996 on various technical and health care aspects
of advanced treatment for wastewater reuse.
The Italian investigation, carded out by means
of a 100 m3/h disinfection pilot plant purposely
designed, built and operated at West Bail (S. Italy)
municipal wastewater treatment plant (3000 m3/h),
was specifically aimed at comparing pathogen
removal, disinfection by-product formation and
costs of 3 altemative disinfectants, namely UV rays,
peracetic acid and ozone. For each disinfectant,
the influence ofwastewater quality was investigated
by disinfecting 3 different municipal effluents,
namely secondary (II), following activated sludge
oxidation and sedimentation, clarified (CL),
following also post-precipitation with aluminum
polychloride, clarified-filtered (F), further submitted
to sand filtration.
The general results of the investigation as well
as those concerning peracetic acid and ozone dis-
infection have been already published [8-11].
Partial results of the UV investigation referring
to bacterial inactivation effectiveness have been
described in a previous note [12].
This paper reports further on UV disinfection
of clarified and clarified-filtered municipal effluents.
The investigation was carded out at the disinfecting
doses necessary for achieving the Italian microbial
standard for unrestricted agriculture reuse
(2 CFU/100ml of total coliforms, based on the
well known State of California Wastewater Recla-
mation Criteria, 1978) with specific objectives of:
• evaluating the effect of UV disinfection towards
selected parasitic pathogens often occurring
in municipal wastewater (Nematodes eggs,
Giardia lamblia cysts, Cryptosporidium
parvum oocysts)
• searching for eventual UV promoted DBP
formation (i.e., nitro-phenols and N-nitroso-
amines)
• drawing economic estimates.
 L. Liberti et al. / Desalination 152 (2002) 315-324 317

2. Materials and methods
The investigation was carried out by means of
the 100 m3/h pilot plant (Fig. 1) with appropriate
configuration for comparingthe performance of UV
radiation on 2 different municipal wastewater
effluents, namely clarified (CL), discharged
directly from WBMP and clarified-filtered (F),
obtained submitting CL to pressure sand filtration
(MF) in the pilot plant. UV disinfection occurred
in a non-contact apparatus (UVA), wherein the
water flows inside Teflon tubes surrounded
externally by low pressure Hg vapor lamps.
The comparison was performed at UV doses
required for meeting the Italian microbial standard
(2 CFU/100ml of total coliforms) previously found
to be 100 and 160 mWs/cm2 for feed F and CL
respectively. On the contrary, the max dose achiev-
able in the conditions investigated (430 mWs/cm2)
was not enough to meet the standard with feed II,
even if the total coliform value achieved (5 CFU/
100 ml) was very close to the target. Further
details on pilot plant, feed characteristics and main
results of the previous part of the UV investigation
may be found elsewhere [12].
For both feeds F and CL, about thirty cycles
(i.e. a given feed submitted to a given dose) were
replicated in the same conditions in order to check
for selected pathogenic parasites (Nematodes
eggs, Giardia lamblia cysts and Cryptosporidium
parvum oocysts) before and after disinfection as
well as for potential DBP formation (i.e., nitro-
phenols and N-nitroso-amines).
Analytical procedures were according to Stan-
dard Methods [ 13] except as specified below:
• Giardia lamblia cysts and Cryptosporidium
parvum oocysts: the method (Standard Method
No. 9711 B as modified by Portincasa [14]) in-
volves pressure (4 atm) tangential ulaafiltmtion
of a 10-1 sample through 142 mm diameter
(1.2 ktm porosity) cellulose acetate membranes.
The membranes were eluted wilh 0.1% Tween 80
solution using magnetic stirring and the eluate
was centrifuged at 1500 rpm using plastic tubes.
The centrate was purified by Percoll-sucrose
gradient and identified by microscopy using
immunofluorescent monoclonal antibodies.
• Nematodes eggs: the method (not standardized
yet) involves the filtration of a 2-1 sample,
membrane elution and centrifugation as for

p2 ¸

~ 7

UVCP

,tl..- UVA

t V

Fig. 1. Pilot plant configurationduring UV disinfectionexperiments(MF, multilayerpressure filter; RV, 5 m3 fibre-glass

vessel; UVA,non contactTeflonUV apparatus; UVCP,UV controlpanel;P, pumps;FM, flow meter;V, valve).
318 L. Liberti et al. / Desalination 152 (2002) 315-324

Giardia and Cryptosporidium. The centrate • liquid chromatograph/mass spectrometer (LC/

was identified by phase- and differential-inter- MS) mod. API 300 by Perkin Elmer.
ference-contrast optic microscopy.
• Nitro-phenols: liquid-liquid extraction GC/MS
method (Standard Method No. 6410 B) modified 3. Results and discussion
for extraction and concentration procedures:
3.1. Parasite removal performance
a 2-1 sample, previously filtered on 0.45-mm
cellulose nitrate membrane, was acidified at As said, several cycles were run at UV dose of
pH 2 with sulphuric acid (1:1) and extracted 100 and 160 mWs/cm 2 for F and CL feed res-
by vacuum filtration on a previously con- pectively to evaluate UV effectiveness towards
ditionated polystyrene-divinil-benzene(SDVB) selected pathogenic parasites (Giardia lamblia
membrane. The membrane was eluted with cysts, C r y p t o s p o r i d i u m p a r v u m ocysts and
methylene-chloride, the eluate was concen- Nematodes eggs) likely to occur in local waste-
trated (up to 1 ml) under vacuum and analysed water and are reportedly resistant to chemical
by GC/MS. Control o f extraction and concen- disinfectants.
tration procedures was obtained with blanks The occurrence of these pathogens in CL and
and standard solutions. F feed before and after UV disinfection is reported
N-nitroso-amines: liquid-liquid extraction GC/MS in Table 1 (average values).
method (Standard Method No. 64 10 B) modified Nematodes eggs were never found in the feeds
for extraction and concentration procedures: admitted to disinfection, confirming the effective-
a 2-1 sample was alkalinised to pH 11 with 10N ness of clarification and filtration steps in removing
NaOH, filtered on 0.45-ktm cellulose nitrate consistently such large and heavy parasites [ 15],
membrane and extracted by vacuum filtration but not towards smaller ones like Giardia cysts
on a previously conditionated C 18 membrane. and Cryptosporidium oocysts detected in appreci-
The membrane was eluted with methylene- able number in feed CL and removed only in part
chloride, the eluate was concentrated (up to by filtration, as expected [16]. In these conditions
1 ml) under vacuum and analyzed by GC/MS. UV radiation was rather effective in both feeds
Control of extraction and concentration pro- towards both protozoan parasites with an average
cedures was obtained with blanks and standard removal around 60 and 65% for Giardia and
solutions. Cryptosporidium respectively.
The following analytical instruments were used:
• tangential ultrafiltration apparatus mod. Sarto- Table 1
con 2 and mod. Sartocon Mini by Sartorius; Selected parasites before and after UV disinfection of
• optic microscope (direct light, phase- and dif- clarified(CL) and clarified-filtered(F) feeds (UV dose: 160
ferential-interference-contrast)mod. Axioskop and 100 mWs/cm2 respectively)
MC 80 by Zeiss;
• fluorescence microscope mod. BH2 by Olym- Parasite Feed In Out % removal
pus; Nematodes CL 0 0 --
• gas chromatograph/mass spectrometer (GC/ eggs (N/l) F 0 0 --
MS) mod. Saturn 3 by Varian with purge and Giardia lamblia CL 345 156 55
trap autosampler mod. 3000 by Tekmar; cysts (N/I) F 114 44 62
• vacuum concentration apparatus mod. AES Cryptosporidium CL 23 8 65
1000 by Savant; Parvum oocysts (N/I) F 6 2 67
 L. Liberti et al. / Desalination 152 (2002) 315-324
These results came not unexpected as protozoan
cysts are resistant species, requiring higher UV
doses than bacteria and viruses [17]. According
to literature data, UV doses lower or similar to
those investigated (around 60, 80 and 120-
180 mWs/cm 2) are claimed to produce 80, 90 and
99% inactivation of Giardia cysts [18-20] and
Cryptosporidium oocysts [21,22] in drinking
water. The multiple barrier concept involving
clarification and filtration (plus eventually GAC
adsorption) prior to UV disinfection undoubtedly
appears the most effective approach for complete
parasite removal in water and wastewater treat-
ment [20].
It must be pointed out that the proved occurrence
of some protozoa in the disinfected effluents does
not cause restriction for their reuse in agriculture.
In fact, according to the WHO guidelines [23], the
only parasites of concern are intestinal nematodes
(MAC < 1 egg/l), never found during this investi-
gation. However, these latter are intended to serve
as indicator organisms for all parasitic pathogens
and it is implied that all helminth eggs and protozoan
cysts should be removed to the same extent to
avoid the risk of waterborne disease outbreaks.
3.2. Disinfection by-products formation
As for eventual formation of UV disinfection
by-products during drinking water production, it
is generally assumed that moderate ultraviolet
irradiation (<50 mWs/cm z) affects the structure
of organic compounds to a much lesser extent than
chemical reagents like chlorine or ozone [ 17]. How-
ever, some results dealing with degradation of pesti-
cides and other specific compounds in industrial
wastewater under high UV doses (> 1000 roWs/
cm 2) indicated the need for further information
about UV promoted transformation of organic
compounds in municipal wastewaters [24,25].
On the basis of photochemistry fundamentals,
in fact, it cannot be excluded that UV irradiation
ofwastewater could affect the identity of the organic
substances through either direct or indirect
319
interaction forming potentially toxic by-products
[26, 27]. In the former case, a molecule known as
a chromophore may be chemically modified as a
result of direct radiation absorption. Indirect
photolysis may occur when UV radiation acts on
a species known as a photosensitiser which
strongly absorbs the radiation energy and the
resulting highly energetic species interacts with
another molecule producing a chemical trans-
formation [28,29].
Considering that amino- and phenolic-deriva-
tives (chromophores) as well as nitrate/nitrite ions
and humic materials (photosensitisers) are species
commonly occurring in municipal wastewater and
potentially capable of reacting [30], nitro-phenols
and N-nitroso-amines were specifically searched
for in this investigation as possible DBPs following
UV irradiation.
Figs. 2-5 show the corresponding GC/MS
chromatograms ofF and CL feeds before and after
UV disinfection (doses of 100 and 160 mWs/cm 2
respectively).
The nearly total overlapping of all spectra
seems to exclude the formation of both harmful
N-derivatives above the instrumental detection limit
(0.01 ppb) in the conditions investigated.
A possible explanation could be the formation
of non-volatile DBPs (NVDBP) which are not de-
tected by GC/MS but only by LC/MS (or MS/MS)
analytical technique. A special experiment was
carefully planned to check this hypothesis at lab
scale. In particular, in order to maximize the pro-
bability of detecting the eventual NVDBP formation,
a 500-ml sample of CL feed was irradiated in a glass-
batch-UV reactor using purposely an extremely
high UV dose (25,000 mWs/cm2). Samples before
and after the irradiation were concentrated 25
times by lyophilization and post-column injected
into a LC-MS spectrometer. The obtained mass
spectra reported in Fig. 6, once again, do not show
any significant difference between irradiated and
non-irradiated samples, excluding the eventual
formation of NVDBPs even under the extreme
irradiation adopted during the experiment.
320 L. Liberti et al, /'Desalination 152 (2002) 315-324

u |

1200

Fig. 2. Nitro-phenols search: GC/MS spectra of F feed before and after LJV disinfection (100 mWs/cm~),

I
fl~ 1200 16~1~ 20~ 2,400

Fig. 3. N-nitroso-amines search: GC/MS spectra o f f feed before and after UV disinfection (100 mWs/cmZ).

.h
! • , !
° I f J
9~ t~9 t500 tfl(~ 2186 240~

Fig. 4, Nitro-phenols search: GC/MS spectra of CL feed before and after UV disinfection (160 mWs/cm2).
 L. Liberti et al. / Desalination 152 (2002) 315-324 321

, ,, | | '-- !

I ' ' I ' I = '' '' - -- ' ' '

800 :12,00 1600 2000 2400

Fig. 5. N-nitroso-amines search: GC/MS spectra of CL feed before and after UV disinfection (160 mWs/crn2).

,1
5,416
4,Se6

$.6d
I 30~.1
2.7d"
.8d -

g.otS,

I
1"~o 2110 310 41o 490 5110 11,30 TOO

GeS" 171,3

I
5t6"

~" 4elll,
d , :'4S.3

z** zlo ~io 4/0 410 s~ ~ Tc~ ,7*

m/I,

Fig. 6. Post injection LC/MS spectra of iyophilizated samples of CL feed before and after UV irradiation (25,000 mWs/cm~).

The above evidences are in good agreement 3.3. Cost estimates

with the conclusions reported for similar investi-
gation stating that UV-promoted transformation To assess the economic feasibility of UV disin-
of chemical compounds, at least in clean waters, fection treatment for wastewater reuse in agriculture,
requires much higher doses than necessary for operation and maintenance (O&M) costs were
municipal wastewater disinfection [31-35]. preliminarily estimated on the basis of the experi-
322 L. Liberti et al. / Desalination 152 (2002) 315-324
mental results obtained by reference to the optimal
UV dose for each feed that permitted the achieve-
ment of the target total coliforms standard of
2 CFU/100ml, i.e., 100 and 160 mWs/cm 2 for F and
CL feeds respectively.
The following assumptions were made:
• O&M costs account essentially for electric
power consumption and lamp replacement,
including also maintenance requirements and
miscellaneous equipment repair costs
• power consumption of UV equipments is
3.1 kWh
• average electricity cost is t~ 0.065/kWh
• UV lamp (t~ 45 each) replacement is based on
8760 h of use.
As shown in Table 2, O&M costs of UV ad-
vanced disinfection of the intermediate (clarified,
CL) and the full tertiary treated feed (clarified-
filtered, F) amounted to t~ 35 and t~ 17.5/1000 m 3
respectively. The above estimates do not include
capital costs and can be influenced by variables
such as feed quality, plant configuration, plant size
(scale factor) and market situation.
Cost effectiveness of UV advanced disinfection,
in particular o f F feed, is evident considering that
chlorination of municipal wastewater just for sea
discharge in compliance with Italian regulations
(20,000 CFU/100 ml for total coliforms) at West
Bari treatment plant costs approximately ~ 5/
1000 m 3.
4. Conclusions
The experimental results of a 9-month pilot-
scale (100 mVh) investigation, carried out at the
West Bari (S. Italy) municipal wastewater treat-
Table 2
Cost estimates for UV disinfection of CL and F feeds at West Bari pilot plant
Feed UV dose, Total coliforms target achieved,
mWs/cm2 CFU/100 ml
F 100 I 6.7
CL 160 1 13.5
ment plant, focused on parasite removal and
disinfection by-product (DBP) formation during
the UV disinfection of clarified (CL) and clarified-
filtered (F) secondary effluents at doses (160 and
100 mWs/cm 2 respectively) necessary for achieving
the total coliforms standard of 2 CFU/100 ml for
unrestricted reuse of wastewater in agriculture,
provided the following indications:
1. At the above doses, UV radiation was rather
effective in both feeds towards protozoan parasites
like Giardia lamblia cysts and Cryptosporidium
parvum oocysts (approx. 60 and 65% removal
respectively); Nematodes eggs were never found
in the feeds admitted to disinfection because already
removed by clarification and sedimentation;
2. The multiple barrier concept involving
clarification and filtration prior to UV disinfection
was confirmed to be the most effective approach
for complete parasite removal in wastewater treat-
ment;
3. None of the N-derivatives (i.e. nitro-phenols
and N-nitroso-amines) searched for after UV dis-
infection of both feeds was detected by GC/MS
analytical technique whilst LC/MS analyses
excluded also the formation of non-volatile DBPs,
suggesting the absence of detectable photo-
chemical reactions at UV doses usually used in
wastewater disinfection;
4. O&M costs of UV disinfection averaged
t~ 17.5 and t~ 35/1000 m3for F and CL feed res-
pectively.
Further investigation is planned to assess UV
effectiveness towards viruses, to exclude UV
promotion of other DBPs, to prevent effluent
recontamination (bacteria photoreactivation by
cell repair and regrowth) and to evaluate possible
O&M costs, t~/1000 m 3
Electric power Lamp replacement Total
10.6 17.3
21.3 34.8
 L. Liberti et al. / Desalination 152 (2002) 315-324
synergy and/or catalytic effects o f UV radiation with
long-acting chemical disinfectants such a s H20 2.
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Environment Federation, Baltimore, Maryland, 1998,
p. 137.