~ Pergamon Wal.Sci. Tech. Vol. 32, No. 12, pp. S9-72. 1995.

Copyright @ 1996 IAWQ. Published by Elsevier SCience Lid

Printed in Great Britain. All nghls reserved.
0273-1223I9S S9'SO + 0'00
PII:S0273-1223(96)OO 139-4

CHEESE WHEY AND CHEESE FACTORY

WASTEWATER TREATMENT WITH A
BIOLOGICAL ANAEROBIC-AEROBIC
PROCESS
F. Malaspina, L. Stante, C. M. Cellamare and A. THehe
ENEA. DivisioneTecnologie di Depurazione e Trattamento Reflui;
Via Martiri di Monte Sole, 4, 40129 Bologna. Italy

ABSTRACT
Research on the anaerobic treatmentof raw cheese whey started in 1990 with the objective of developing a
technologysuitable for medium size cheese factories that have growing disposal problems and cannot afford
high investmentcosts for whey valorisation technologies (such as whey protein and lactose recovery. spray
drying. ete.), In order to couple process stability and high loads. a new downflow-uptlow hybrid reactor
(DUHR) has been designed. The reactor was able to reach B, values around 10 g CODeI-I.d- I, with 98%
COD convertedto gas and effluentsolubleCOD valuesclose to 1,000ppm; no external additionof alkalinity
is required to maintain a stable pH thaI was constantly around 6.5-6.7 in the downtlow pre-acidification
chamber and around 7.5 in the bio-methanation uptlow chamber. The high strength of the cheese whey
treated gives an effluent that still contains high amounts of COD, ammonia nitrogen and phosphorus and
therefore a post treatment is required in order to meet standard limits. Tests of post treatment were carried
out during two years with a Sequencing Batch Reactor (SBR). The SBR was tested at various FIM values
with different durations of anoxic-anaerobic-oxic cycles, obtaining. under certain conditions, more than
90% removal of COD. nitrogen and phosphorus. Copyright @ 1996 IAWQ. Published by Elsevier Science
Ltd.

KEYWORDS
Anaerobic digestion; cheese whey; downflow-upflow hybrid reactor; two-phase anaerobic digestion;
Sequencing Batch Reactor; nutrient removal.

INTRODUCTION
Milk whey is a byproduct that the dairy industry can reprocess or transform into other valuable products.
Small and medium size cheese factories, like most of those in Italy which produce high quality typical
cheeses, do not have the economic convenience or the market dimension to afford most of the reuse
technologies available. Traditionally, these cheese factories are strictly connected to pig farms where milk
Whey is directly used as the liquid basis for animal feed.

If for any reason (environmental, sanitary, economic) the downstream animal farm is lacking, a solution to
Whey disposal must be found. This is the case of most cheese factories of the Sardinian Pecorino cheese;
OWing to epidemic disease, pig farming in Sardinia was drastically reduced a few years ago. From the
59
60 F. MALASPINA et al:

analysis of the problem that ENEA was called to do, anaerobic digestion - followed by aerobic post-
treatment together with process wastewater - came out theoretically as the most convenient solution. Biogas
produced could be economically reused to substitute most or all fossil fuels used for process thermal energy
generation; moreover, anaerobic digestion should destroy the greatest part of COD, allowing the effluent to
be post-treated together with the process wastewater in the existing treatment plants of the factories with
only little modifications to the plant structure. Anaerobic treatment of whey is not a new process. The
literature about it is very rich, but most of the authors have worked with very diluted whey, a much simpler
waste to treat. Because of its very high biodegradability (close to 99%) and concentration (-70 geCOD-l-l)
and the very low bicarbonate alkalinity (-50 meq-l-l), raw whey is a particularly difficult substrate to treat in
high loaded anaerobic digesters. Moreover, granulation is not supposed to occur using whey as substrate
(Hickey et al.; 1991), making difficult the use of UASB reactors.

In the present paper, part of the results of laboratory experiences started in 1990 are reported. The research
was first carried out on a hybrid upflow reactor, later on a two-phase system and finally on a downflow-
upflow hybrid reactor, a new concept that was specifically developed for raw whey treatment. The main
concepts behind this design are that phases are separated within the same reactor and that, in order to obtain
more stable process conditions and higher resistance to shock loadings, a downflow completely mixed pre-
acidification zone is followed by a plug-flow upflow biomethanation zone. The CSTR zone acts as a shock
buffer. In fact, the influent is introduced at the top of the downflow chamber, where mixing is very high and
bacterial activity should be correspondingly high. This design could reduce the risk of pH drop in case of
recycle pump failure. Moreover, the DSFF design of the pre-acidification chamber should reduce the
passage of acidifying biomass to the upflow part.

The effluent from the anaerobic reactor cannot be discharged into surface waters before a post-treatment for
the removal of residual organic matter and nutrients. Its composition is highly unbalanced, because most of
the organic matter has been destroyed in the anaerobic step, while nitrogen and phosphorus are
quantitatively conserved as soluble inorganic compounds. A post-treatment has been therefore designed and
tested. The process is based on a Sequencing Batch Reactor. that from previous experiences has
demonstrated to be very suited to treat wastes containing high nutrient concentrations. The organic matter
needed for the biological removal of nutrients is provided by the addition of a synthetic stream of cheese
factory process wastewater that presents a very high carbon/nitrogen ratio.

MAlERIALS AND METHODS

All experiments were carried out in mesophilic conditions, utilizing raw whey as substrate, obtained from a
cheese factory nearby Bologna, derived from the manufacturing of cheese from 90% cow and 10% sheep
milk. Its main characteristics are reported in Table I.

Table I. Raw cheese whey characteristics

Parameter Unit Average St. Dcv.
Total COD rng- I-I 688 J4 11518
Soluble COD mg-I" 57876 11272
TSS g-kg" 1.3 1.14
VSS g-kg" 0.94 0.74
TKN mg-l" 1462 263
NH4+-N rng-l" 64 31
Total P mg-I" 379 49
P04-P mg-l" 326 64

Data from previous experiences (Malaspina et al.; 1994) were used for the design of a downflow-upflow
hybrid reactor (DUHR) that is still in operation in our laboratory (Fig. 1); results from the first nine months
are presented here. The reactor, of the total volume of 51 1 is composed of a downflow pre-acidification
chamber where influent is pumped, filled 2/3 with a channelled polyurethane filter; on the bottom of it, this
 Cheesewheyand cheesefactorywastewater treatment 61

chamber is connected with an upflow one - arranged as a hybrid reactor - that contains a similar filter
located in the top 2/5 of it. The volumetric ratio between the two parts of the reactor is 1:5. A recycle from
the top of the second chamber (r~.5) is applied to provide alkalinity and dilution to the influent.

An SBR of 5 dm 3 of maximum capacity (Fig. 2) operated with anoxic. anaerobic and aerobic phases for
each cycle in order to obtain biological removal of organic substances and nutrients. Several tests were
carried out at different loadings and operating cycles. in order to evaluate the best operational condition for
COD. nitrogen and phosphorus removal.

In order to decrease the very high TKN/COD ratio of the anaerobic effluent, a synthetic stream of cleaning
waters of the cheese factory. rich in organic substances and poor in nutrients. was mixed with the anaerobic
effluent in a ratio similar to the true wastewater/whey volume ratio of the Parmigiano Reggiano cheese
manufacturing process: two parts of cleaning water and one part of digested effluent.

Effluenl

Final Volume

Initial volume

pHi i

Fig. I. The DUHR reactor Fig. 2. The SBR

Analyses of COD. TS. VS. TSS. VSS. NTK. total P and sulphides were performed following the Standard
Methods (APHA. 1989). Monovalent anions and cations were analysed using a HPIC (Dionex 4000i). Total
alkalinity (TA) was measured titrometically at pH 3.8. Bicarbonate alkalinity (BA) was calculated from
VFA and TA values. VFA were determined gas chromatographically. using a DANI 8510 GC equipped with
a 25 m - 0.53 mm 1.2 mm capillary wide bore column (Alltech SO FA bonded FSOT) and a FID detector;
hYdrogen was used as carrier gas and 2. 2. dimethylbutyric acid was employed as internal standard. Biogas
Was metered with wet tip gas meters and analysed gas chromatographically with a DANI 3865 GC equipped
with a TC detector; a custom made 6m - 3.18 mm teflon packed column filled with Chrompack Hayesep C
allowed good separation of N2. CH 4• C02 and H2S,

RESULTS AND DISCUSSION

Results of the test carried out on DUHR are presented in Figs 3 to 15. After 40 days needed for the
adaptation of the seed sludge taken from a pig waste digester. By was gradually increased up to the design
loading rate of 10 g COD-I-I_d-I.
62 F. MALASPINA ~t al.

1~ - ~oo
I)

III
150 - I -.
~
.,-
-:;

.: "- ..; I t
h
&
E 100 1,- I
~ 7 A
If _A
1:::

-
~
~
-c
:J..
:-
50 ~-
• .-I'" r_' ~• _.
_
..t-tr,
• --" I
It
h-
Yt. ~"A'
d ?! I

II I ,
tu "-
I I I I I I I I I I I 0 I
I

I I
~
I I
- I I , I I I I
" , I

day day

Fig. .l Bv applied 10 DUHR Fig. 4. VFA in acidogenict a) and Fig. 5. pH of acidogenic (A) and
methanogen ic (0) section rnethanogenic (0) section

After three weeks at high load, some methanogenic biomass washout due to the very high mixing caused by
gas production and by the passage of acidogenic biomass from the first into the second chamber resulted in
growth of VFA concentration and particularly of acetic acid, without causing therefore any unbalance of pH
in the upflow chamber. On the other hand, the increase of loading rate resulted sometimes in pH decrease in
the downflow chamber. Phase separation allowed us to maintain stable pH in the methanogenic section also
after an accidental recycle breakdown happened during a weekend around day 215; during this accident, pH
in the first chamber fell down to 4.9, while in the second chamber it didn't show any significant decrease.
The process recovered very rapidly after recycle restore and a very little alkalinity supply (5.25 meq .l"
Na2C03 + 2 meq-r! CaC03). Recycle ratios in the range 1:4 to I:2.5 were usually sufficient to provide
alkalinity and dilution to the influent. No external alkalinity was used for pH control, other than after the
accident described.

COD removal was always well over 90%; within the experimental By range, no dependence of COD
removal on By is shown. During a 36 days period of stable operation at 10 g COD.I- 1od-1, 98.4% COD
removal (Fig. 13) and methane production of 0.33 nl-g CODin- 1 (Fig . 14) have been calculated. Peak biogas
productions close to 10 volovol-lod- I were achieved at the maximum loading. Methane content in biogas
averaged 53%, with no significant dependence on By. The high gas flow rates resulted in upflow velocities 2
orders of magnitude higher with respect to the liquid upflow velocity (Fig . 15). Due to the little sludge yield
and to the absence of granulation. the correct design of a glVs separation device is critical in order to
maintain the reactor biomass content at high loading rates.

During some steady loading periods at different By, intensive profile sampling allowed us to calculate some
basic process parameters. reported in Table 2. Calculations were done making the assumption, derived from
literature data on the polyurethane support material used (Van Rompu and Verstraete. 1988), that the filter
contained 15 g vss-i-t. Data are in very good agreement with the ones obtained in previous experiences
with a two-phase CSTR.

During the whole experiment the internal reactor liquor always showed a viscous consistency. This
phenomenon was particularly relevant during period of overloading, like for instance in the load increase
periods of the start-up phase, when severe washout can occur because the biomass settling capacity is highly
reduced. This phenomenon is probably due to the over-production of ExoPolySaccharides (EPS) of bacterial
origin; their synthesis uses mono- and di-saccharides, glucosamines and uronic acids as substrates, and is
favoured by high concentrations of sugars and by high CIN ratios (Sutherland, 1985; Guiot et al., 1988).
 Cheese whey and cheese factory wastewater trealment 63

180
160
.. .
140 , '?
H.OO
~ 120
't 0 -:
,0 ,
t 100 , &J 2n , ~ 6.00
E "'5
J.
..,«'' ,
80 ~ 4.00
-c
,,
.. ~~
u,

..... > 60
40
20
0
.t • o·

Ih~ B
0 ' ;:
~

2.00

I I
12 0 4 6 8 10 12 o~ ~ ~~ 8 ~ ~ :i~~~ :; ~
- - - - - ""1 t"~ N N
Bv gCO D.L·1.d· 1 day

Fig. 6. pH vs. Bv in acidogenicf a ) Fig. 7. VFA V5 . Bv in acidogenic(. ) and Fig. 8. Biogas ( +) and methane (0)
and mcthanogen ic (0 ) sect ion of rnethanogenic (0) section of DUHR product ion of DUHR (normal ised volumes)
DUHR

SIXXI -r- - - - - - - - - - - - - ---, 6000 ..,-- - - - - - - - - - - - ,...-,

7 (J<X) l'
I SOOO
(JO< XI
~ 4000
:..,; 50m
"',
E ~I XX)
00
E 3000
-c
11. .1000
~ 2000
>
21XX)
1000
IOOll
() + ...,..l. ......--,.....~;,a.. o --t-,.~ ......~;W~ f!1J
=N ~ ~ ~ 8 g ~ S ~ ~ ~ ~ ~ 80 100 120 140 160 180 200 220 2-10 260
day
day

Fig. 9. Concen trat ion of ma in VFA in acidogenic section of Fig. 10. Concentration of main VFA in rnethanogeni c section
DUHR (acet ic acid (+), propionic acid ( e). n-butyric acid of DUHR (acetic acid (+). propionic acid (e). n-hutyric acid
(A), n-vale ric acid ( +) and n-capronic acid (0 ) (A ), n-vuleric acid ( +) and n-capronic acid (0)

Table 2. DUHR kinetic parameters

Period Bv FIM Uspec . Yo!l s. J.I

(d) (gCOD·L,J·d ") (gCOD·gVSS" ·d") (gCODr·gVSS-' ·d") (gVSS·gCODr") u l ')

7 7.5 0.58 0.56 0.079 0.044

7 9.5 0.63 0.56 0.032 0.019
9 10 0.66 0.55 0.060 0.033
7 8 0.66 0.51 0.120 0.066
average 0.63 0.55 0.075 0.()4
51. Dev. 0.04 0.03 0.04 0.01
64 F. MALASPINA et al:

.. ..
1.02 ..,...- - -- - - - - -- , I . ll ~

1-
•• •• "
::; • •....• •
~ 0 ). - " 1l.9S
•• •
' -. Ii •
~
-; • •
~ 0 .96 _ . • • • ll.% • • ••
::
e (} .() .J-
•
•
•
• ~
~

(5
O . 9~
•
•
• • •
1l .'1~ -
,i,
• U 1l.'12
• • •
ll.'l
II ~ I> II I 12

uay Il l' ~ CO D 1.·' u·1

Fig, I I. COD removal during DUHR rest Fig. 12. COD removal of DUHR as
function of applied B v

I~I K K I .1I KWI ..,...- - - - - - - - - ----,.,

Slo",' = O') X ~
Slop e = 1l . J .1 ~
121KKI
H I\ ~ =0. ( )t)<) R 2 = 1l1)HH
A

IllI KK) -
21KK I
s: SI KKI
~

,.. C 151M I :J
(,IKM I ~

- )(KWI
~I K K I

~( K k) 51WI

= '"
x
r,
x
x
:;
x
x
<0
s
,:;..
x
:::'"
x
x
c.
X
"-
=t
= '" x
.. '" .. :r,
x
c o
x
;;,
x
=t
X xx
x
~
x
;::..
"
x
:z
"
'"
x
~

~ CODIIl
~ COD IIl

Fig 13. T\bss of COD rcm versus mass or Fig. 14. Volume of methane produced from
CODin Juring a period of 36 d at B v or 10 inlet COD during the same period.

gCOD·L-l.tl- 1 in DUHR.
10 ..,...- - - - - - - - - - - - - - - -,

-= 0.1
-
:: 11111

II I WII
o
o

= c. -r ::!: x " r; =t z x i='xo c.

r: • =t ~
c. c'
d;l~

Figure 15. Liquid flow in acidogenic (.) and methanogenic(.) section of reactor and gas flow in both sections
(0) of DUHR during tbe experiment

Effluent characteristics are summarized in Table 3. The high total COD values are to be ascribed to the
relevant bacterial washout at high loads - also witnessed by the high TSS and VSS figures - due to the
absence, in the laboratory reactor, of a well designed gNs device. The effluent soluble COD was very low
with respect to the influent values but not enough to approach effluent standards, which were also exceeded
by nitrogen and phosphorus. It is therefore necessary to provide a post-treatment before effluent discharge.
 Cheese wheyand cheesefactorywastewater treatment 65

Table 3. DUHR effluent characteristics

Parameter Unit Average St. Dev. sample n.
Total COD mg·r i 5839 3117 25
Soluble COD mg-l" 1085 1263 25
TSS g-kg" 5.19 3.30 18
VSS g-kg" 3.02 1.69 14
NH;-N mg-I" 816 122 6
POrP mg-I" 35 21 3

The addition of process wastewater deriving from dairy machinery washing, which presents high CIN ratios,
allows us to compose a wastewater that can be biologically treated. Laboratory experiences carried out using
a Sequencing Batch aerobic reactor have already demonstrated the feasibility of combined carbon, nitrogen
and phosphorus biological removal. SBR operated with different cycles in order to test the best operational
conditions for removing both organics and nutrients. The cycles are summarized in Fig. 16 and the average
main characteristics of influent for SBR treatment, made of one parts of effluent from DUHR and two parts
of process wastewater, are reported in Table 4.

Table 4. Main characteristics of SBR influent during the tests

Parameter Unit Average value
l
Total COD mg·r 4048
Soluble COD mg-l" 2611
TKN mg-I" 435
NH/-N mg-l" 165.27
TSS g-kg" 1.45
VSS g-kg" 1.08
Total P rng-I" 82.42
P04-P mg-l" 27.76

..---......- Tesl2

--.....-
Wlthelr• •

Tesl3

--
..-

Tesl.

Tesl5

II I I I Ii i i i Ii i i i "i "i
i i i i .6 i I I 12i i i '4 i i i 18i i i dili I'd! I i ell! i pal

Figure 16.SBR cycles for the five test periods.

66 F. MALASPINA el al:

The first test lasted three months with 4 NitrificationlDenitrification (N/DN) cycles/24h at loading condition
as showed in Figs 17 and 18 and an average F/M of 0.215 gCOD.gVSS-I.d- 1 (corresponding to HRT of 2.5
days and B, conditions of 0.93 gCOD.dm- 3.d-I).

The reactor gave very good performance both in COD and nutrient removal as shown in Figs 19 and 20.
Overall performances are summarised in Table 5 (concentrations are in mgol-I, Relative Standard Deviation
(RSD) and removal rates are in percent).

Table 5. Inlet and outlet average value during test #1

Parameter inlet value RSD outlet value RSD removal
Total COD 3841 19 311 42 90.7
Soluble COD 2672 22 137 40 94.1
TKN 410 16 16 141 93.2
NH/-N 166 42 0.49 172 99.6
NO,'-N 0.83 80 33 88
Total P 66 46 9,63 23 93.2
PO"-P 18.6 66 4 34 78.79

5000 -r----::::-----,-- - .,---..,.----,--r--r- r 800 04 5 -:r-,..---,----,.---..,.---- - ---,

.1500, 700 04
.1000
600 ,,- ~ 0.35
.r 3500 E -en 0.3 +--# -++-i--''----,-~---_i
~ 3000 500 E: (/)
~025 ,
8 :?500 400 0 0
o 8 02 +--+---+--"'..
o 2000 -1"'-H -~-r--i--T---Trb 300 ]I
~0. 15
~ 1500
200
'5
~
0
0.1 -'j- -+--+-!-- i- - -i--1-'-=---1
1000 -1========='i:=;= === ===±=f.
500 ~ 100 0.05+--,--"-~-+-­
o 0 0 +--+-..-r...r- , -""""-f-.-"T"'",,,,--,---1
o 10 20 30 40 50 60 70 80 90 100 o 10 20 30 40 50 60 70 80 90 100
Day Day

Fig. 17. COD cone. in SBR during test #1 Fig. 18. F/M applied during test # I

0.95
09
iii 085
~g 0 9 + lr_--~- \
>
g 08
c: '" 0.75 -:f-<-----,--->- y .- ;-
c:
0 85 0 .7 -:J----;--~J' --+-~-­
0 65

0 6 +--+--i--i---,-.....,.....--,r--.,---.------i
o 10 20 30 40 50 60 70 80 90 o 10 20 30 40 50 60 70 80 90
Day Day

Fig. 19. COD removal in SBR during test #1 Fig. 20. Nand P removal during text # 1
 Cheese whey and cheese factory wastewater treatment 67

The averaged values show quite high COD and nitrate effluent values, but they are due to some peaks that
don't represent the normal performance, when values very close to the Italian effluent standards were
reached. Very good removal of phosphorus was obtained particularly at the end of the period, when stable
biological P removal was established. The sludge had an excellent settleability.

In order to try to improve reactor performance, a second test was carried out with shorter and more frequent
N/DN periods (6 cycles/24h) with the following operating conditions: By of 0.86 gCOD.dm· 3.d-l, HRT of
3.6 days and FIM of 0.128 gCOD.gVSS-I.d· l . Results were not satisfactory, as summarised in Table 6
(concentrations are in mg-ll, RSD and removal in %), and the test was stopped after 48 days.

Table 6. Inlet and outlet average value during test #2

Parameter inlet value RSD outlet value RSD removal
Total COD 3241 42 1259 62 61.1
Soluble COD 2015 74 210 54 89.5
TKN 385 10 65 141 83.1
NH;·N 121 79 17.6 156 85.4
NO;·N 0.72 75 60.7 62
Total P 28.8 6.45 76
PO.·P 13.24 79 5.5 43 58.4

Outlet data showed high values of ammonia and nitrate due to low rates of nitrification and denitrification,
whereas the very high value of COD was caused by washout of sludge due to the very consistent presence of
filamentous bacteria.

The third test was run with 5 N/DN cycles/24h with organic loading as reported in Fig. 22 (averge By of
0.94 gCOD.dm- 3.d-l, HRT of 4 days and FIM of 0.188 gCOD.gVSS-I.d· I). Under these operating conditions
and cycles (Fig. 16) good nitrogen removal with low concentration of ammonia and nitrate in the effluent
was obtained, while P, TKN and CODt removal were poor due to wash-out of high SVI sludge (Figs 23 and
24). A summary of reactor performance of this test is reported in Table 7 (concentrations are in mgel· I, RSD
and removal are in %).

I -e- CO Ol inret II -- CO Ol outle t I

5000 2500 0.45 -,-- ,--- -- ----r- - - ---,
I 1 )1 /1 0.4 +-~f_-:...-_i_-+--- ,t -_i
4500
--:.., 4000
)\1 V r I!f J' 2000 -:; -; 035 -'}----'I -_ _ -'--+_ -;-_ /
63500 / i\ I j i V '\ 6
a ~ 0.3 +-+-----+--j-~-I ·- \ -_j
I '\. V'T I \ 1500 o C/)
83000
00
.s 2500 I t "'I / I -
III I I I
~ r
]?
~

g
0.25 4--
02 4--
; -- -
.--::"- -.:-
-;-- i-
-1--1-
-
-7-/
-/

]? 2000 1000
.5 1\ II I I so ~ 0. 15 + - i-- - \--+-.... -H- - _j
o 1500 o
8 1000 \ I II i 500 8 ~ 0.1 +-i---\r--j--~-
500 I \ II I I 0.05 +-i----;--j-~---1
o I II I .: I V
o o 4-- r--r--r----i- --r- .--i
o 10 20 30 40 50 60 70 o 10 20 30 40 50 60 70
Day Day

Fig. 21. COD concentration in SBR in test #3 Fig. 22. F/1\1 applied during test #3
68 F. MALASPINA et al.

Table 7. Inlet and outlet average value during test #3

Parameter inlet value RSD outlet value RSD removal
Total COD 3542 21 800 106 77.4
Soluble COD 2271 62 194 113 91.43
TKN 434 51 148 55 65.7
NH4+-N 136 68 4.21 186 96.8
NO,'-N 0.43 140 5.4 100
Total P 141 45 49 70 65
P04-P 23.4 84 7 34 70

1. 1
I
I I I
0 - '-
o.
0.9
Q ~ I I I 0 -
0 8-+--~P'
,7 ~ I I I
'~E" . -I- -i--
07 H
_ 0.8
~
@0.7 f I "J I II
I
8 0 6+-~1---i-Cl--!- \ OJ
- 0.6 ~ I :"'1 I I
u
05 0.5
I I I
h i I -
0 .4

03 ~"'9="T"""""F--r~,;-"..-...r--l
0.4

0.3 I
1:- I
,
Tot Nrem

TOC al P rem

I I
~
Ii
I
i
'r-,
II
~
i
o 10 20 30 40 50 60 70 o 10 20 30 40 50 60 70
Day Day

Fig. 23. COD removal in SBR during test #3 Fig. 24. Nand P removal during test #3

The fourth test lasted 210 days with 4 NIDN cycles/24h at the operating conditions reported by Figs 25 and
26 (average value of B, 1.36 gCOD.dm- 3.d-l . HRT 3.68 days and FIM 0.27 gCOD.gVSS-I.d- I). Reactor
performance is reported in Figs 27 and 28, and summarised in Table 8 (concentrations are in mg-I- I, removal
and RSD are in %).

Table 8. Inlet and outlet average value during test #4

Parameter inlet value RSD outlet value RSD removal
Total COD 4480 44 1082 99 77.8
Soluble COD 3031 67 201 105 93.3
TKN 484 12 153 III 68.3
NH/-N 267 46 II 239 95.8
NO,'-N 1.19 1.56
Total P 109 85 44.4 113 59.3
PO.-P 59 103 7.49 90 87.3

Also in this test some bulking problems (Fig. 29) caused wash-out and high values of total COD, TKN and
total Phosphorus in the effluent (low removal as shown in picture); despite the high load, good performance
was obtained for nitrification and denitrification (see graph of Fig. 28).
 Cheese whey and cheese factory wastewater treatment 69

10000 1.2.,.--:--:-- - - - --:--:-----,----,

8000 _ 1 + + + --:--l-+ -,-__'

.:- '9
~
0 ~ 08
0 en
U >
S
0>

4000
b
o
06

~
o

Day Day

Fig . 25. COD conce ntration in test #4 Fig. 26. F/M ap plied during text #4

1.2 ...,----r-- - -,....- -.,...-.,--.,-- - ---,

1. 1 +-+---,--;--i .~_t__:_--
0.9
1

..
~ 07
08 ~ 0.9 ..:1:.-:f-,-!-.:-,~ -+~~, t_--__i

.,g 08+'\"1-~7- r+:' ;I-~~----i

~ 0.6 a: 0 .7
o ~ 0 .6
8 0.5 tt-trt±±±±:±:±:ji l1 z'" 0.5
04
04 4-- -""Il-"- ,' -'-i
0.3 + I" ---rl 0.3 l-l-,=;= c=;;=-= ==-="-
0.2 +:-;-'++H+~r::+~ 0.2 ++-r~-r-r-r-+_r_.___r---j
o 0
g 8N ~ o 0 0 a 0 0 0 o 0 0 0 0 co 0 N oq to a:) 0 N
.. ID N .. ID
N N N

Day Day

Fig. 27 . COD removal in SBR dur ing test #4 Fig. 28 . N and P remova l durin g lest #4

-~ 5 +-++-+~_t_~_:__+_t·t __j
til
~ 4 +-- -l-+-+-+-+-- -l---:'--iH
~

§ 3 +t_+,--l-+~~--.---:-- I -j
u
>-
~ 2
;;
:S1
o

8 ~
Day

Fig. 29. Outlet suspended solids du ring lesl #4

In order to remove filamentous bacteri a, a fifth test was carried out with unm ixed fill at high load (B ; of
1.28 gCOD.dm- 3.d- l , HRT of 3.66 days and F/M of 0.266 gCOD.gYSS ·I .d· l ) and 3 NIDN cycles/24h (see
Fig. 16) according to "K, strategy" in bulking control (Chudoba, 1985).

NST l /· 12·0
70 F. MALASPINA et al:

The test lasted 76 days and gave very good results in filamentous bacteria control: after 60 days only few
filaments were found in the sludge that, on the other hand, was very rich in big sarcinas similar to G-
Bacteria. Good performance on COD and nitrogen removal was obtained (Fig. 32), while phosphorus
removal was poor (Table 9), probably due to conditions that promoted the growth of G-bacteria instead of
polyphosphatic ones (Cech and Hartman, 1993).

Table 9. Inlet and outlet average value during test #5

Parameter inlet value RSD outlet value RSD removal
Total COD 4737 30 1960 61 58.6
Soluble COD 3064 41 375 T2 87.7
TKN 463 6 157 61 66.1
NH/-N 136 54 6.13 122 95
NO'-N
~
1.56 57 0.78 80
Total P 65.65 36 43 96 34.8
P04-P 24.42 87 8.41 liS 65.5

80oo -:r- .- r== = = = = " ,

7000 -,1
. --+---1 __ 0.25
-c; 6000 i--t~i1-"";--r---,---rf--rH
g 5000 4. --hH--+--t-----j:---~O-+-H '"
~
en
02 -t-~-t-" +--+---+--;-- +--1
g. 4000
>
g o,5 -t---+-~-+--"<-i--+---+-+--j
o
:: 3000 +-t-JIU--+- +--:::P-'l ~ 0 1 -+---+- -'--l-- i-'>"
8 2000 + --l;{- + ""'i-- +---+--t-\--+--i ::;
u: 005 + -+__;--+_.l---+'\.~I /_~
, 000 +---+- +--+- +--+---+- '''P'\,-j
o-+.-...-r+.,.,.,..,,..;...-....-+"TTTi-rr--+rrrri~r+r-.,..,..j o-+---+""""'+-"-+-""""""'-+"-""""r--""....--rl
o 10 20 30 40 50 60 70 80 o ' 0 20 30 40 50 60 70 80
Day Da y

Fig. 30. COD in SBR during test #5 Fig. 31. F/M applied during test #5

0.9 I I ;1-
08 I....... J /_
_ 07 4- -
I - i -- -+-I r,-
- -;-- I;,)

~ I I I /\ C
06
G." I i 1- l!J
~ 0 5 +-- -"-,:iI-- - :---_t-I-- _i
g 04 "';--s. I
+---.;-.:;~~!__-_tI l--_i
I
U
I ~
0 3 ir=="'===~"'-o..;;::::_:t--_j
II
0.2 'i -B- fla COOI ~--
0.1 ..j -.- Ela COOs
I
o +::::;:::;:::;:::::;=;:::::::=;'..,..-,...__~..,...,...j
o 20 40 60 80
Day

Fig. 32. COD removal in SBR during test #5

In summary. the SBR demonstrated to have a very good potential in nutrient and organic removal. In order
to obtain good nutrient removal together with high sludge settleability (low SVI), an equilibrium between
number of NIDN cycles, their duration and feeding modalities has to be found. As a general remark, short
and frequent NIDN cycles theoretically allow lower nutrient concentrations to be reached, but too short
cycles can negatively affect nitrification and denitrification. Short cycles probably favour filamentous
 Cheesewheyand cheese factorywastewater treatment 71

bulking; fast and unmixed feeding and longer oxic/anoxic-anaerobic time ratios can be promising strategies
of bulking control.

Phosphorus removal was checked during tests by fractionation of cellular P in samples taken at the end of
anaerobic and aerobic phases. During the aerobic phases the content of polyphosphates in sludge (15-20
mglgVSS) was about four fold respect to the samples taken during anaerobic phases (3-5 mglgVSS) and
therefore it is possible to state that biological phosphorus removal is involved in the process. Nevertheless a
high fraction ofP was removed by precipitation as insoluble "metal-bound" compounds (20-50 mglgTSS).

Organic matter removal was very good but it was reduced by wash-out due to filamentous bulking and by a
background of unbiodegradable COD. Moreover the tests show that also in presence of very clear
supernatant, a relatively high amount of COD was found in effluent This condition is caused by a
background of unbiodegradable very fine suspended-colloidal COD coming from anaerobic digestion. For
this reason the effluent from SBR needed a tertiary treatment to remove the unbiodegradable COD. This
Conditionoccurs only when the cheese factory uses very low amount of water (and therefore discharges low
amount of cleaning-waters) like in our simulation; in many cases however, the volume ratio between
cleaning water and cheese whey is from 4 to 6, with a dilution effect that has a positive effect on effluent
quality.

Most of residual colloidal and soluble organic substances, that are not biologically degraded, can therefore
be easily removed by chemical flocculation and physical separation (flotation or settling) .

Coagulation and flocculation tests were carried out dosing FeCl 3 (200-400 mg Fe.g CODin'l) and anionic
polyelectrolyte (7-20 mg polyelectrolyte.g CODin·l) with very good results: 80% COD removal and 90%
total P removal.

In conclusion the biological anoxic/anaerobic/oxic process is able to treat successfully effluent from
anaerobic digestion of cheese whey when this is mixed with factory cleaning-waters. Physical-chemical
treatments can be required in order to reach standard values when low amounts of cleaning waters are
discharged from the cheese factory.

CONCLUSIONS

Despite its very high biodegradability, raw cheese whey is a quite problematic substrate to treat
anaerobically, because of the lack of alkalinity. the high COD concentration, the tendency to acidify very
rapidly, the difficulty to obtain granulation and the tendency to produce excess of viscous exopolymeric
materials of probable bacterial origin that severely reduces sludge settleability and can be a cause of biomass
washout

The DUHR design demonstrated COD removal rates of 98% at considerably high B, (10 g COD-I-I_d,l)
without any need of external alkalinity supply. The particular design allows us to obtain phase separation in
the same reactor, therefore reducing investment costs with respect to separated reactors.

The SBR is a very flexible reactor for both organic matter and nutrient removal from wastewater and is
Particularly suitable for post-treatment of digested whey.

The residual unbiodegradable colloidal and soluble organic substances, that are not degraded biologically,
can be easily remove-d by chemical flocculation and physical separation (flotation or settling).

ACKNOWLEDGEMENTS

Authors wish to acknowledge Emanuele Sbaffi and Chiara Parmigiani for their precious collaboration .
n F. MALASPINAet al:

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