ttuIer Rt,scar~k Vol 15, pp 361 tO 373 (K~43-13M'81 '030361-1350200"0

C Perglmon Press Led 1981 Printed m Great Britain

ACTIVATED SLUDGE RESPONSE TO EMULSIFIED

LIPID LOADING
STEVE E. HRUDEY
Department of Civil Engineerinf. University of Alberta.
Edmonton, Albert& Canada

(Received September 1980)

Abstract--The possible adverse effects of natural fats and oils (lipids) upon biological wastewater treat-
merit systems have been the subject of much speculation and some disagreement. This study addressed
the specific question of possible effects of emulsified lipids upon activated sludge process performance.
Lipid form and fatty acid composition were determined on wastewater samples. Experimental studies
employing bench scale activated sludge units indicated that effluent quality is either not significantly
different or occasionally is better than that from sucrose supplemented control plants receiving compar-
able organic loading (F/M < 0.80). Furthermore, emulsified lipids clearly do not exhibit an inhibitory
effect on either the specific soluble substrate utilization rate or the mixed liquor oxygen consumption
rate.
However. et~uent quality parameters were found to be dependent upon unit lipid loading to a highly
significant degree. As well, unit lipid loading (8 lipid g-J MLSS-day) provided a better explanation of
effluent quality than either the fond to microorganism ratio or the mixed liquor lipid burden (g lipid
extracted from MLSS g-~ MLSS). The findings of this study indicate the conditions where lipid over-
loading occurs. This data provides a basis for controlling emulsified lipid loading to the activated sludge
procc&s which should apply to plants which receive high lipid loading such as those encountered in
industrial wastewater treatment applications.

INTRODUCTION used to refer to non-hydrocarbon lipids as predomi-

A major fraction of the organic content of domestic
wastewater may be attributed to "oils and greas~"
according to several investigators (Mahlie, 1 9 ~ :
Painter & Viney, 1959; Viswanathan et al., 1962:
Hunter & Henkelekian, 1965; Loehr & Kukar, 1965;
Zweig et al., 1967: Loehr & de Navarre, 1969; Far-
rington & Quinn, 1973: Given et al.. 1974; Fedorak,
1975; Lordi & Lue-Hing, 1976; Waggon & Britcher,
1976). The significance of "oils and greases" in waste-
waters has been the subject of extensive debate. The
current investigation was aimed at one component of
that debate, namely the response of activated sludge
treatment performance to emulsified lipid loading.
The "oils and greases" term has generally referred
to that material which may be extracted from a waste-
water sample by a nonpolar solvent such as hexane or
freon. This determination may include both mineral
oils and greases {hydrocarbons) as well as animal or
vegetable oils and greases. The general biochemical
term "lipids" may be interpreted to include some of
the hydrocarbons; namely, those which can be com-
monly biosynthesized. However. in the context of
wastewater terminology, lipids are commonly inter-
preted as representing natural fats and oils which are
predominantly composed of fatty acids, glycerides
{esters of fatty acids with glycerol) and phosphogly-
cerides (esters of fatty acids and phosphoric acid with
glycerol combined with other radicals). Several other
classes of lipid compounds are known but these are of
limited quantitative significance with respect to waste-
waters. For present usage, the term "lipids" will be
36!
nantly represented by fatty acids and glycerides.
The debate concerning the significance of lipids in
domestic wastewater has been partially clouded by a
failure to distinguish hydrocarbons from non-hydro-
carbon lipids when discussing oil and grease. As well,
confusion has arisen between strictly physical prob-
lems in sewerage and treatment systems and possible
adverse effects upon treatment process performance.
Conventional municipal biological wastewatcr
treatment normally includes some form of primary
treatment, often with skimming, Such treatment
should effectively remove floatable lipids prior to the
biological process. Consequently. any adverse lipid
effects upon biological treatment performance would
be expected from emulsified rather than floatable
iipids.
The biodegradability of lipids has been evaluated
by several investigators (Loehr & Roth, 1968; Mala-
n e y & Oerhold, 1969: Novak & Kraus, 1973;
Kramer, 1971). Loehr & Roth (1968) found that the
biodegradability of long chain fatty acids increased
with decreasing chain length and increasing degree of
unsaturation. Malaney & Gerhold (1969) found long
chain fatty acids to be very resistant to oxidation by
activated sludge microorganisms. Novak & Kraus
(1973) determined the maximum substrate removal
constants for fatty acid biodegradation to be an order
of magnitude less than for other biodegradable sub-
strates. Kramer (1971) proposed that the hydrolysis of
lipids to release free fatty acids was a potential rate
limiting step in the biodegradation of lipids by a
batch biological treatment process. These studies aug-
 362 STEVE E. HRUDEV
gest that llpids, while clearly biodegradable, may be
less amenable to microbial degradation than other
substrates.
Lipids, fatty acids in particular, have been reported
to be inhibitory to various bacteria by several investi-
gators including Nieman (19541, Smith & Alford
{19661, Ferdinandus & Clark (1969L Kahn & Katamay
( 1969~ Dutton & Evans (1970), Hedgecock (1970L Gal-
braith et al. (19711. Steffen & Calvin (1971L Kabara et
al. (19721, Kondo & Kanai (1972L Levis,an (19721, Sheu
& Freese (19721. Sheu & Freese (19731, Freese et al.
(1973L Galbraith & Miller {1973a, b, cL Fay & Fanas
(1975L Butcher et al. (1976L Kondo & Kanai (19761
and Miller et al. (19771. The common features appar-
ent from these studies are the general susceptibility of
gram-positive and gram-negative bacteria to short
chain fatty acids but a general resistance of the latter.
with a few exceptions, to longer chain fatty acids.
Concern over the behavior of lipids in biological
treatment systems has resulted in the evaluation of
both full scale (Loehr & Kukar, 1964: Loehr & de
Navarra, 1969; Willoughby & Patton. 1968; Paulson
et ,d.. 1971: Mulligan & Sheridan, 1975; Lordi & Lue-
Hing, 1976: McDermott, 1976; Young. 19791 and
bench scale {Hydroscience. 1971: McCarty et al..
1972; Anderson et al.. 1973: Lordi & Lue-Hing, 1976l
activated sludge performance in treating lipid bearing
wastewaters.
Many of these studies have concentrated on evalu-
ating lipids removal by activated sludge. Performance
in this regard is generally good. The question of poss-
ible lipid effects upon other indicators of activated
sludge performance has received less attention.
Results which are reported do not provide a dose-
response basis to either conclusively demonstrate any
influence of lipids upon process performance or to
account for lipid loading in activated sludge process
design or operation.
Overall. the literature appears to offer some contra-
dictions. It is apparent that lipids are readily removed
by activated sludge and that they are clearly bio-
degradable, although at perhaps a slower rate than
other substrates. Clear evidence of lipid inhibition of
bacteria has been demonstrated with individual
species but such effects have not been unambiguously
addressed for the activated sludge process. Evidence
of lipid effects upon activated sludge treatment per-
formance, as measured by other efltuent quality par-
ameters {BODs suspended solids), has generally been
inconsistent. Where poor performance occurred, rele-
vant factors such as process organic loading have not
been elaborated.
The regulatory response to the contradictory infor-
mation on lipid effects has been varied. Many munici-
palities enforce sewer use bylaws restricting grease
discharges (which may or may not be distinguished
according to hydrocarbon character} with regulated
values ranging from 25 to 300 mg l - L. The Technical
Practice Committee (WPCF, 1973) recommended the
removal of floatabte lipids at source. However, for
emulsified lipids they recommended: "'If oils and
greases are biodegradable and in a physical state that
does not cause clogging or undue maintenance prob-
lems in the wastewater facilities, the discharge of these
substrates can be accepted in a wastewater treatment
system." EPA (1975k in assessing the need for munici-
pal sewer system pretreatment standards, rec-
ommended that oils and greases of animal or veg-
etable origin be subject to no limitation. Thus,
although many municipalities maintain bylaws limit-
ing lipid discharges to municipal sewers, the current
trend appears to be away from regulation of lipid
discharge.
Since cause-ffect lipid loading design information
for the activated sludge process is limited and the
regulation of emulsified lipid discharges to municipal
sewers may have significant economic consequences,
this investigation was aimed at assessing the effects of
emulsified lipid loading upon the activated sludge
process.
EXPERIMENTAL PROCEDURES
Analytical
Wastewater and etfluent analyses were conducted, as far
as possible, by Standard Methods IAPHA, 1975L The stan-
dard procedures and modifications, where applicable, are
summarized in Appendix [.
Determination of lipids was subjected to considerable
evaluation M o r e methods were adopted. The lipid content
of mixed liquor solids was determined by method 502 D
tAPHA, 1975). Lipids in liquid samptMs were ultimately
determined by a partition gravimetric procedure em#oy-
ing continuous liquid-liquid extraction with certified ACS
petroleum ether as solvent. This procedure, described in
detail by Hrudey (1979). was found to be demonstrably
superior to the partition gravimetric (method 502 A) and
soxhlet extraction procedures (method 502 C) described by
Standard Methods (APHA, 19751.
Fatty acid composition of lipids was determined, follow-
tng extraction as above, by transesterification with BFj:
methanol as described by Metcalfe et al. t 19661. Semi.quan-
titative results were obtmned by spiking individual sam#es
with heptadecaaoic acid prior to methylation. Gas
chromatographic analysis was performed using flame ion-
ization detection and a 2m x 6ram x 2ram i.d. #ass
column packed with 50 DEGS-PS on Supelcoport
100/120 at 150~C and 50ml rain -x nitrogen career gas
flow.
'Vlonit oriny
In order to specify relevant experimental conditions for
the study, a limited monitoring program was performed to
consider the fatty acid composition of common industrial
sources of lipid bearing wastewaters and the form and fatty
acid composition of lipids in primary sewage at a full scale
activated sludge plant.
The fatty acid composition of meat packing industry
wastewaters was evaluated with a pmr of sampl,~ obtained
from the City of Edmonton industrial waste sampling pro-
gram. Each sample was a combined composite of the emu-
ents from all four of the major meat packing plants in the
city. The oil seed industry wastewater evaluation was based
on a composite sample from a rapeseed processing plant.
 Activated sludge response to emulsified lipid loading 363

Table I. Fatty acid composition of wastewaters

Average composition* (2o by weight)

C12:0 C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 C20:1

Meat packing plant

wastewaters 4.9 6.0 34.8 2.2 16.5 30.4 5.2 --
(City of Edmontonl
Rapeseed processing -- -- 15.3 -- 4.9 58.8 ! 8.0 0.8 2.1
Plant wastewater
Combined fatty acids -- 3.4 21.4 6.2 10.1 43.1 13.1 2.7
Edmonton primary,
sewage
Free fatty acids -- 3.0 26.4 7.7 20.4 34.3 7.1 1.0
Edmonton primary
sewage

* Cx:y x = number of carbon atoms; .v = number of unsaturated bonds.

The distribution of fatty acids between free fatty acids
and combined fatty acids (glyceride esters and/or com-
pound lipids) was evaluated by obtaining five daily grab
samples of primary effluent from the Edmonton Gold Bar
Wastewater Treatment Plant. Each sample was subdivided
into five aliquots which were separately extracted for gly-
cerides, compound lipids and hydrocarbons under basic
conditions followed by extraction for free fatty acids under
acidic conditions.
The fatty acid composition of the wastewaters is sum-
marized in Table 1. The distribution of fatty acids between
free fatty acids and combined iipids is summarized in
Table 2.
Bench scale studies
A modified version of the bench scale activated sludge
unit described by Black (1968) was developed for the ex-
perimental evaluations on activated sludge. The units, illus-
trated in Fig. I, were designed to operate with the aeration
basin as a complete mix reactor. This mixing regime was
verified by a step dye input test of the complete mix model.
The experimental set up in the laboratory was arranged
with four plants in adjacent chambers of a common, insu-
lated temperature control bath. Bath temperature was con-
trolled at 20 ~ lcC with a reczrculating 500 glycol water
solution. The field investigation was conducted at ambient
temperatures, ranging from 21 to 26~C.
Inf]uent substrate was supplied from a pair of covered
125 I. polyethylene tubs (one control one lipid dosed). Sep-
arate feed lines of teflon tubing were provided to each unit
with the feed solution drawn by a multiple head peristaltic
pump operating on silicon tubing segments. Effluent from
each unit was collected in a separate calibrated polyethy-
lene tank.
Based on the findings summarized in Tables 1 and 2, a
test lipid emulsion was prepared using 50% free fatty acids
and 50:o ether extracted beef suet. The higher proportion
of combined fatty acids than observed in the monitoring
study was chosen to compensate for the possibility that
primary sewage samples had suffered additional hydrolysis
during the 2 h which elapsed between sample collection
and analysis. Fatty acid composition was set on the basis
of the three predominant fatty acids observed in the moni-
toring program: palmitic (16:0k stearic (18:0) and oleic
(18:1). The emulsion was prepared according to the pro-
cedures described by Hrudey (1979). The result was found
to he stable for 2-3 days before any visible signs of fat
separation occurred.
Laboratory studies were conducted with two control
units and two lipid dosed units. Lipid dosed units received
the basic synthetic sewage (Table 3J plus the lipid emulsion.
Control units received the basic synthetic sewage plus a
supplement of sucrose, calculated to approximate the or-
ganic loading (based on BODs) of the lipid dosed units.
Field studies were conducted at the Red Deer Waste-
water Treatment Plant with two lipid dosed units, one su-
crose supplemented control and one plain primary sewage
control. Primary sewage was collected in one daily hatch
to prepare the feed solutions. The lipid emulsion prepared
with primary sewage was sampled at the beginning and
end of 24 h with the average analysis being used to calcu-
late the lipid feed concentration. Loss of lipid over 24 h
averaged 13.4 + 0.9%. The sucrose supplemented control
was also calculated to match the organic loading of the
lipid dosed reactors but sucrose was added to the reactor
by a separate ~ t r a t e d solution feed in order to avoid
ral~d degradation of sucrose in the primary sewage reser-
voir. The plain primary sewage control operated on unmo-
dified primary sewage to provide operating data relative to
the full scale plant.
Operating conditions for the bench scale reactors are
summarized in Table 4. The high aeration rate was used to
ensure complete mix conditions in the aeration basins.
Mean cell residence time was controlled by daily batch
wastage of mixed liquor from the aeration basin.
Process evaluation
In order to consider the effects of lipid upon activated
sludge process performance it was desirable to consider
relevant process loading parameters. Mulligan & Sheridan
11975) and Hydroscience (1971) proposed limiting the unit
grease loading to activated sludge plants. Their proposed
loading parameter, analogous to the food to micro-
organism ratio, was calculated as g of lipid applied per g of
mixed liquor solids per day and was abbreviated for this
study as Lipid/M.
Table 2. Lipid form in Edmonton primary sewage
Giyceride and compound
lipid fatty acids
plus hydrocarbons Free fatty acids
Sample day (wt To of lipids) (wt % of lipids)
1 30.3 69.7
2 44.7 55.3
3 28.8 71.2
4 25.4 74.6
5 44.4 55.6
Mean 34.7 65.3
 364 STEVE E. H R U D E Y

Adiustable Baffles --,....~.

,[
I

Aeration Chamber
$etthng
Chamber
/7
."
, : " / ~ Effluent
:" / z Outfall

!, ,~ Teflon CoateO , / /
." ,~$ur/aces~
(, //
\ ', //
' \ \,
\\ //
\~\'., Flow Owerter- a //
k
:o ~ " - - - 7 ~ PerforateO 1/8"
( / Tubing for Aoration

Fig. 1. Bench scale activated sludge unit.

Previous investigations (Hydroscaenc. 1971 : McCarty et Liquor solids, was determined and was abbreviated as ML-
al., 1972) had not explicitly di~mksed the food to m~cro- Lipid.
organism ratio (F/M) as an explanatory factor for observed
effects o1" $rea~ upon aetivauat sludl~ pcrforrmmce. Or-
ganic Ioadiltti was deemed important and consequently RESULTS AND DISCUSSION
F/M was determined for all p r o a ~ data.
Finally, Hydro~iance (1971)speculated that process fad- The treatment performance data was evaluated for
ure occurred when mixed liqour solids beemne excessively frequency distribution and all parameters were ob-
load__
~_ with grease causing der,r t u e d specific gravity and
sludF scttleability. As well Banjeri et al 11974) speculated served to exhibit skew distributions. A K o l m o g o r o v -
that greue coating of maxed liquor may interfere with the Smirnov test was performed on each parameter to test
ability of the floe to absorb oxygen resulting in impmred the suitability of a log normal disu'ibution for descrip-
aerobic nmmbolbm. If either mechanism was important for tion of the data distribution. This hypothesis was ac-
the caste of emulsified lil~ls and activated slud$, the liptd
ceptable at the 5% significance level for all parameters
content of the mixed Liquor should exert some control over
proceu pcrfornlanc~ Consequently, the mixed liquor lipid except for total organic carbon and sludge volume
burden, calculated as g of lipid extracted per g of tmxed index. However, the log normal hypothesis could be
accepted for these parameters at the 2% significance
level and a log normal distribution was adopted as
Table 3. Basic synthetic sewage fced the model distribution for all parameters.
The process loading and performance data for both
Concentration the laboratory based synthetic sewage studies and the
(50 I. batch)
ling l -I ) field based primary sewage studies are s u m m a r v , e d in
Stock solution
Tables 5 and 6. respectively. Geometric means are
FcC'ls (0.25 gl- ~) 1.5 reported in keeping with the log normal model.
Urea (50 gl- t) t0 Despite-attempts to match the BUDs of the control
K zH PO,--crystal 42 influents to those for the lipid dosed influcnts, com-
NH,Cl---~rystal 134
parison of the experimental results suggests that the
Bactopeptone ( Difco k---crystal 100
Sucrose-----trystal 20 lipid dosed plants were exposed to higher F / M values
CaCI2 (30.5 gl- t) 12 overall. The significance of the differences in mean
MgSO, 122 gl- i) 16.5 F M values between the lipid dosed and control
Typical Edmonton tapwater analysis
Table 4. Bench ~cale activated sludge reactor operating
Cations Anions conditions
milli-cquivalents I- l milli-equivalents I-
(rag l - I ) (mg 1- t) -~eration basra hydraulic retention
time 8 h
Ca" ~ 0.90 (18.0) HCO3- 0.72 (43.9) Aerat=on rate 70 m z m - J
Mg=" 0.55 (6.6) SO~" 1.39 (66.7) Mixed liquor suspended solids 1000-3000 mg I
Na" 0.75 (17.3) CI- 0.09 43.21 Mean cell residence time 7.5 days
 Table 5. Summary of treatment performance with bench scale reactors for synthetic sewage experiments

Effluent Emuent Effluent

total Etguent Effluent total filtered Effluent
suspended total filtered organic organic ether
Lipid/M ML-Lipid F/M Solids BODs BODs carbon carbon extractibles
(gg-1 d-*) (gg I) (gg-= d -I) (mgl 'l) (mgll (mgl-I) (mgl-J) (mgl-i) (mgl-l)
Control
Median 0.61 5 l0 7 II 6 3
Geometric mean 0.61 6 13 8 10 7 3 3
Range 0.26-1.68 <1 263 4.495 3-375 3-250 3 190 <19
No. of observations 60 60 58 51 58 58 57
Lipid dosed
Median 0.31 0.061 0.92 8 21 12 18 I0 7 e"
Geometric mean rs
0.28 0066 0.96 II 23 12 18 I0 6
Range 0.04-0.78 0.019-O.210 0.45-2.07 1-196 7-167 4-43 7-86 4-31 !-28
No. of observations 67 67 67 64 67 50 67 66 64

Table 6. Summary of treatment performance with bench scale reactors for primary sewage experiments

Effluent Effluent
total Efliuent Effluent filtered Effluent Oxygen Sludge E.
m

suspended total total organic ether consumption volume

Lipid/M ML/Lipid F/M solids BODs organic carbon extractibles rate index
(g g-I d - ' ) (gg-1) (gg-ld-I) (mgl -l) (mgl l) (mgl-l) (mgl-J) (mgl-1) (mgg-1 h-l) lmlg-l)

Control
Median 0.33 18 10 22 18 6 10.7 281 .-i
Geometric mean 0.33 31 18 27 19 5 I1.1 255 qo

Range 0.15-0.08 2-536 2-154 13 202 11-60 1-16 6.2--19.6 68 478

No. of observations 19 19 19 17 17 8 19 19
Lipid dosed
Median 0.17 0.072 0.48 9 7 19 16 5 11.9 225
Geometric mean 0.19 0.078 0.49 I0 I0 24 19 5 12.0 174
Range 0.09~0.50 0.033 0.210 0.25-1.05 I 420 2--154 12-202 11-.60 < 1-44 6.3.22.1 50446
No. of observations 38 38 38 38 38 34 32 38 38 38
366 SIEVE E. HRUDFY

j i I i , l , i i 1 sucrose controls were not significantly different

2.00 ./ (P > 5%) based on effluent total suspended solids.

total BOD~, filtered BODs and filtered organic car-
bon. Effluent total organic carbon was significantly
IP < 20) higher for the lipid dosed plants.
For the primary, sewage experiments, the logarith-

,i / ! mic mean values of effluent total organic carbon, fil-

o 1/ 0.00 0.50
......
I O0 i. 5 0
F / M , g / g - clay
Fig. 2. Spoctfic sutntrate utilization rate as a function of
F/M for lipid dosed reactors.
plants for both the synthetic sewage and primary sew-
age experiments was tested by a two-tailed t-test on
the loga.,'ithmic means. For both data bases, ,=ummar-
ized in Tables 5 and 6, the differences were significant
(P < 5%).
For the single purpose of comparing results from
the lipid dosed and sucrose supplemented controls,
these data sets were truncated at extreme values of
F/M until the Iogmithmic mean F/M values for con-
trol and lipid dosed plants were not significantly dif-
fete=at at the 5% level. This corresponded to an F/M
range of 0.45--0.80 for the synthetic sewage exgen-
merits and 0.25--0.80 for the primary sewage experi-
~'lltS.
For the synthetic sewage experiments, a two taded
r-test, on the truncated data sets, indicated that the
logarithmic mean effluent quality for lipid dosed and
tered organic carbon and mixed liquor oxygen con-
sumption rate were not significantly different
IP > 5;,) between the lipid dosed and sucrose control
plants using the truncated data. However, for effluent
total suspended solids, total BODs and sludge
volume index, the logarithmic mean values for the
controls were significantly higher tto P < 0.1%). This
2.00
; effect was attributed to the dramatic adverse effects of
filamentous microorganism growth in the sucrose
supplemented control.
These results indicate that for moderate orgamc
loadings (F/M < 0.80) a carbohydrate such as sucrose
will cause as much, or more, upset of activated sludge
performance as a corresponding organic loading of
emulsified lipid.
Further evidence regarding activated sludge process
response to emulsified lipids relative to sucrose was
obtained by determining the specific substrate re-
moval rate, U. This parameter, calculated as g of sol-
uble BODs removed per day per g of mixed liquor
solids, was compared to process loading determined
as F/M and as Lipid/M (for the lipid dosed reactors~.
For this purpose, the lipid emulsion was considered
as soluble BODs. The comparisons of U with F;M
for the lipid dosed and sucrose control reactors for
synthetic sewage experiments are plotted in Figs 2
and 3. respectively, The comparison of U with
l.,ipid/M is plotted in Fig. 4.
Considering Figs 2 and 3 first, the relationships are
essentially identical as indicated by the linear regres-
sion line fits. Furthermore. Figs 2 and 4 clearly dem-
onstrate that there is no indication of any inhibitory

, * 0 t ; ; : :

I
2.00 200 4 =.4.

>, 150

I
1 o0 l!

5 t
0.50 ~. 050

f
o.oo j 0 00 I
000 0.50 ~ O0 1'50 2100 O0 02 04 06 08

F / M , g i g - clay Lipid/M, g/g - day

Fig. 3. Spec:fic substrate utilization rate as a function of Fig. 4. Specific substrate utilization rate dependence upon
F/M for sucrose supplemented control reactors. unit lipid loading (Lipid/MI.
 Activated sludge response to emulsified lipidloading 367

Table 7. Significance level of regression relationships between performing and loading parameters
for synthetic sewage experiments
Loading parameters
Lipid/M ML-Lipid F/M F/M
Performance parameters (lipiddosed) (lipiddosed) (lipiddosed) (sucrosecontrols)
Total suspended solids 0.1?.g 0.100 0.100 100
Total BODs 0.10'o 0.1 ?o 0.1 ?~ > 5~"
Filtered BODs > 50o" > 5o* > 50o* > 5:o*
Total organic carbon 0.lo > 5.g" 5% I~o
Filtered organic carbon 0.Io 50 > 5,/" 5~
Ether extractibles 50 > 50~* > 5//o* N/A
" Not significant.

effect on soluble substrate removal as a result of
emulsified lipid dosage. If such had occurred, some
levelling of the relationships should be observed at
higher lipid ioadings. Such levelling is clearly not
indicated by the experimental results.
Evaluation was performed to determine which, if
any. of the process loading parameters influenced
treatment performance. For the log normal data, this
was achieved by performing linear regression analyses
on the log transformed data and testing the hypoth-
esis that the slope of the regression line was signifi-
cantly different from zero (Sokal & Rohlf, 1969).
Lipid/M, ML-Lipid and F/M were each separately
tested as the independent variable.
The results of the significance testing on the regres-
sion analysis are summarized in Table 7 for the syn-
thetic sewage experiments and in Table 8 for the
primary sewage experiments.
These results indicate several highly significant
(P < 0.1g) relationships between performance par-
ameters and the process loading parameters. The sig-
nificance level indicates the percentage probability
that the slope of the regression relationship appears
to he non-zero clue to random chance rather than due
to the actual presence of a linear dependence. All sig-
nificance tests were evaluated for the actual signifi-
cance level demonstrated by the data beyond a signifi-
cance decision level of 5,0. With this approach, type
II error is based on the commonly accepted 50 sig-
nificance level since rejection of significance was only
concluded for data not significant at the 5~o level.
However, the actual significance level achieved does
show the relative strength of data for supporting
rejection of the null hypothesis (that no relationship
exists between parameters). The relationships will be
discussed in turn for each performance parameter.
Effluent total suspended solids indicate a highly sig-
niticant (P < 0.1%) dependence upon the lipid load-
ing parameters (Lipid/M, ML-Lipid). The dependence
upon F/M for lipid dosed plants is highly significant
for the synthetic sewage experiments and exhibits a
lesser, but high significance level (1%) for the primary
sewage experiments as do the relationships with F/M
for the control experiments.
Effluent total BODs exhibits highly significant
dependence {P < 0.1%) upon all loading parameters
with one exception. For the sucrose controls in the
synthetic sewage experiments, total BODs fails to
indicate significant dependence upon F/M (P > 5%).
The reason for the discrepancy between the synthetic
sewage experiments and the primary sewage experi-
ments may he due in part to the nutrient medium.
The sucrose controls in the synthetic sewage experi-
ments eventually developed a culture with a slimy,
gelatinous appearance which was quite distinct and
not typical of healthy activated sludge floe. This
occurrence may have lead to some erratic perform-
ance. Although filamentous organisms were observed

Table 8. Significance level of regression relationships between performance and loading parameters
for primary sewage experiments

Loading parameters
Lipid/M ML-Lipid F/IV[ F/M
Performance parameters (lipiddosed) (lipiddosed) (lipiddosed) (sucrosecontrols)
Total suspended solids 0.1g 0.1g 1'o 1o
Total BODs 0.I~0 0.10o 0.10o 0.1g
Total organic carbon 0.1% 0.1% 0.1% 1%
Filtered organic carbon 1~o 1o 5% 5%
Ether extractibles 0.1% 10o 500 N/A
Sludge volume index 0.1% 5g 0.1% 5,/o
Mixed liquor oxygen > 5~," > 50g* > 5~,* > 5%"
consumption rate

* Not significant.
368 STEVEE. HRUDEY

to proliferate in the sucrose controls for the primary over other loading parameters is consistent for the
sewage experiments, the atypical gelatinous sludge did primary sewage experiments.
not occur. This development indicates one of the Sludge volume index was reported only for the
possible pitfalls of using strictly synthetic sewage primary sewage experiments and. again, highly signifi-
experimentation for the investigation of activated cant dependence (P < 0.1%) upon Lipid/M is appar-
sludge performance. ent. along with a similar observation for F M (lipid
Effluent filtered BODs was only monitored for the dosed). Sludge volume index was dependent upon
synthetic sewage studies and is not found to indicate ML-Lipid and upon F'M for the sucrose controls at
significant dependence upon any of the loading par- the 50 significance level.
ameters over the range of values tested. Mixed liquor oxygen consumption rate was
Effluent total organic carbon indicates highly sig- reported only for the primary sewage experiments.
nificant dependence upon Lipid/M IP < O.l'o) for the The finding that this parameter did not depend sig-
synthetic sewage studies. Highly significant depen- nificantly on any of the loading parameters is consist-
dence {P < 0.1"~) is indicated for all loading par- ent with the earlier observation, based on specific sol-
ameters in the primary sewage experiments and with uble BODs removal, that lipid loading does not cause
F:M in the sucrose controls the significance level is any inhibitory effects upon overall activated sludge
l",,. In this case. as indicated earlier, the highly signifi- metabolism.
cant dependence upon Lipid/M. in preference to other Since the process loading parameters themselves
loading parameters, is consistent for both sets of interact, the foregoing findings were evaluated by per-
experimental data. forming multiple power regression analysis of the per-
The foregoing trend continues for effluent filtered formance parameters against all three process loading
organic carbon where Lipid/M offers the most signifi- parameters (Lipid/M. ML-Lipid. F/M). Analysis of
cant dependence relationships for both data bases. the resultant partial regression coefficients, using the
Effluent ether extractibles exhibit significant primary sewage data. indicates that for effluent total
(P < 5,) dependence upon Lipid/M while failing to suspended solids, effluent total BODs and effluent
show significant dependence upon either ML-Lipid or ether extractibles only the partial coefficients for
F;M (lipid dosedl in the synthetic sewage expert- Lipid/M are significant (P < 5%).
merits. The trend of higher dependence upon Lipid'M Overall. the assessment of performance parameter

lOOt
i

'~ Syntnehc
~ e
0 ~ .
EJt~rlmoc~ts
~ h'lrnl~
~m.onl$
"-g~ V

>8

Lto~dlM ML-L,pid F M (L,oid-OoIH~l) #'M IContrOISl

Loading Parameters
30! . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

-~s s0i-
E2
# oc.
8 o
c~ ....
nno=

~. >~c' 5 4C~- --
"SE.
5 ~
J ' > . . . . -
2 ~ 20- ~'~
}. ....

LIDLO/M ML'L;OIO F/M (L;!:h(~'OOll~l~l~l F.'M (Controls1

Loading Parameters
F*g. 5. Coefficient of determinauon by loading parameters for total suspended solids and total BOD~.
 Activated sludge response to emulsified lipid loading 369

1000 ' ,]
qucntly, the monitoring of lipid loading, for activated
800
sludge plants receiving significant quantities of emul-
6OO

400
300
/ sified lipids, should provide a better description of
process behaviour than could be achieved solely from
F/M.
The finding that emulsified lipids loading can
200 explain poorer effluent quality while affecting neither
the specific soluble substrate utilization rate nor the
E mixed liquor oxygen consumption rate suggests that
loo the mechanism of lipid overloading does not involve
"6 80 any metabolic inhibition of beterotrophic bacteria in
co activated sludge.
The influence of Lipid/M upon major effluent par-
"0
e- 4o ameters is indicated most clearly for the primary sew-
(~
Z 30 age experiments. This influence is summarized in Fig.
o') 6 for effluent total suspended solids, Fig. 7 for effluent
03 2C total BODs and Fig. 8 for effluent ether extractibles.
The least squares power regression line is plotted for
each case. The outer interval on each plot reflects the
I--' 10
95"0 confidence region for a single future observation
C
(D .i while the inner interval reflects the 950 confidence
6 " /
region for the mean of future observations at the
given value of independent variable.
An alternate analysis of this data which does not
rely on specifying the functional form of dependency

10OO
''1
800
600
0.00 0.~ 0.2 0.4 0.8 1.0
Lipid/M, g/g-day 400
300
Fig. 6. Effluent total suspended solids response to unit
lipid loading (Lipid/M) for primary sewage experiments. 200

dependence upon loading parameters indicates that

Lipid/M considered alone generally describes process
E
performance better than either ML-Lipid or F/M.
Another means of measuring the degree of account- E3 60
ability provided by the individual loading parameters O
m 40
is the coefficient of determination. This parameter
expresses the fraction of linear variation in the depen- 30
dent variable which may be explained by linear vari- 20
ation in the independent variable. The coefficient of
determination is exhibited for total suspended solids
and total BOD~ in Fig. 5. The coefficient of determi- lo
W
nation indicates that Lipid/M considered alone is able 8
to account for as much as 470 of the variation in 6
effluent total BOD~ (primary sewage studies) and can
generally account for more than one third of the vari- 4
ation in performance parameters. Other process load- 3
ing parameters generally can account for less of the
variation in performance parameters with the lipid 2
dosed reactors.
These findings indicate that, although the influence
1 i,,l.
of emulsified lipids upon activated sludge perform-
0.06 O.1 0.2 0.4 0.6 1.G
ance is undoubtedly related to organic loading, the
unit lipid loading (Lipid/M) provides a more specific Lipid/M, g / g - d a y
indication of process performance than is possible Fig. 7. Euent total BODs response to unit lipid loading
with organic loading based on BOD5 (F/ML Conse- (Lipid/M) for primary sewageexperiments.
370
t /
40 //
_ 30
E
~ 2o
"x 10
W .
~ 8
e- .
E i
.2= 4 ing industries.
2 Bench scale reactors, when receiving only primary
/
i sewage, were observed to perform significantly better
/ than the full scale plant treating the primary sewage
/
i / .'
0.06 01 0.2 0.4 0.6
Lipid/M, g/g-day
Fig. 8. Effluent ether extractibles response to unit hptd
loading ILipid/M) for primary sewage experiments.
is provided tn Fig. 9 for effluent total suspended
solids. Fig. 10 for effluent total BOD+ and Fig. l l for
effluent ether extractibles. The plotted points rep-
resent the geometric means of groupings of 5 or 6
data pairs (effluent parameter and Lipid/M) for
Lipid/M values taken in ascending order. The varta-
bility about each data point represents the geometric
standard error of the dependent variable about its
geometric mean. These summaries indicate that as
Lipid/M increases linearly both the mean values and
the variability of all three effluent parameters in-
crease. Both consequences are clearly undestrable for
the consistent operation of an activated sludge plant.
It is notable that. while effluent total suspended
solids IFig. 91 and effluent total BOD~ IFig. 10) even-
tually reach unacceptable levels, the effluent ether
extractibles (Fig. 11) remain low at even the highest
lipid loading values. This observation is consistent
with the generally accepted concept that acti,,ated
sludge is very efficient for lipid removal. However. tt
also indicates that problems can occur with the re-
moval of other parameters while lipid removal is rela-
tively unaffected.
STEVE E. I"IRUDE
The data summarized tn Figs 9 and 10 indicates
little problem is encountered at emulsified lipid load-
mgs of less than 0.25. This would correspond to a
primary sewage concentration (Lo/ of 250 mg I- ~ for
an activated sludge plant providing an aeration basin
detention time 10) of 6 h and carrying a mixed liquor
suspended solids concentration (X) of 4000mg I -t
based on the general relationship:
L,, = tLipid/M) iX)10).
This data indicates a much higher tolerance of acti-
vated sludge to lipids than was reported by Hydro-
science (1971). However. the difference is likely due to
the use of emulsified iipids in the current study as
compared to direct addition of pure oil in the Hydro-
science study. The results reported here provide a
basis for the design and operation of an activated
sludge process exposed to significant lipid Ioadings.
Such situations will be most commonly encountered
at industrial waste treatment plants for food process-
It must be stressed that the results reported were
obtmned with bench scale reactors receiving sewage
feed consistent in strength over daily test periods.
used as basic feed for the study (Hrudey. 1979). A
scale up factor of 1.3 was deternSined on the basis of
effluent BODs and this combined with an industrial
discharge peaking factor of 2 (for discharge over only
12 h of an operating day) may be used to recommeno
a more conservative limitation of Lipid/M to 0.10.
This conservative value should ensure reasonable pro-
tection against adverse performance due to emulsified
lipids. This recommendation presumes that lipid dis-
charges would be reasonably equalized over 12h
periods. The recommendation also specifies the total
emulsified lipid load received by the plant. Conse-
quently, the numbers reported could apply directly to
an industrial plant treating its own wastewater. Any
application of this data to limit discharges to munici-
pal sewers should account for dilution with domestic
sewage, which in most cases would be considerable.
In many such cases, regulation of emulsified lipid dis-
charges to sewer may well be superfluous. However.
these findings ind.icate that emulsified lipid loading
would warrant attention where the lipid discharge
constitutes a significant proportion of the total sew-
age flow.
CONCLL SIONS
The conduct ol bench scale activated ~iudge expert-
mentation using both synthetic sewage and primary
sewage has led to the following conclusions with re-
spect to emulsified liplds.
I. For moderate organic loadings iF M < 0.801.
effluent quality for lipid-dosed activated sludge plants
 Activated sludge response to emulsified lipid loading 371

130

120 ,

Geomelrlc M e a n of I n t e r v a l
110 1 t R a n g e ot _- 1 GN~of~r~etr~c StanclarO Error

100 -

(3) 90 -
E
5 80 .
._

70 .i
"0
O~

C 60 -

m 50 -

m
"5 40 .
I--
L

30 i
2
W
20 4 I

1o . ][., T

0 ,
0 O0 0 10 0.20 030 0 40

Lipid/M, g/g - day

Fig. 9. Effluent total suspended solids dependence upon unit loading (Lipid/M) for primary scwa~
expenmems.

80 G e O n l e t r r c M e a n of I n t e r v a l
T R a n g e ot _- I G e o m e t r i c Slan~larO E r r o r

70

60

E
=5 50
r',l

03 40

0
I-- 30
r- i
20 b
_=
UJ
"F/
10
T ..... #

0
0 O0 0 ~C 0 20 0 30 0 40

Lipid/M. g/g - day

Fig. I0. Effluent total BODs dependence upon unit lipid loading (Lipid/M) for primary sewage
experiments.
w.= 15 3 - - f
.,7.
20 . . . . .
Geometr, Meart o* InlervaJ
SE glenqe or I Geometrpc S t a n t m t o E,rof
_ t,
l't[. r
~ e
,3 ..........
300 0 10 020 030
bpid/M, g/g - day
Fig. 11. Effluent ether extracttbles dependence upon unit
lipid loading ILipid/M) for primary sewage experiments.
was generally not significantly different from sucrose
supplemented control plants and. in some cases, was
significantly better. The poorer performance of su-
crose controls was attributed to observed filamentous
bulking.
2. Effluent quality parameters depend upon unit
lipid loading (Lipid/M) to a highly significant degree
IP < 0.1"o). Lipid/M provided a consistently better
description of efliuent quality parameters than con-
ventional organic loading (F/ML Based on the data
obtained, monitoring of lipid loading to activated
sludge should provide a more direct process indicator
than F ' M for plants where the lipid content of
primary sewage is high.
3. Emulsified lipids were found to exhibit no in-
hibitory effect upon the activated sludge specific sol-
uble substrate removal rate. U (g filtered BODs re-
moved d a y - i g - t mixed liquor suspended solids).
over an experimental range of Lipid/M from 0.04 to
0.78 g lipid d a y - t g - t mixed liquor suspended solids
based upon synthetic sewage studies.
4. Emulsified lipids were found to exhibit no in-
hibitory effect upon the mixed liquor oxygen con-
sumption rate over an experimental range of Lipid,' M
from 0.09 to 0.50 g lipid day - t g- t mixed liquor sus-
pended solids based upon primary sewage studies.
5. The foregoing observations suggest that lipid
overloading of activated sludge exerts an effect by
some mechanism other than metabolic inhibition of
heterotrophic bacteria.
6. An activated sludge process subjected to lipid
overloading can exhibit poor performance in terms of
general efltuent quality (total suspended solids, total
B O D s l without necessarily indicating poor remo,,al
of lipids. This observation confirms the ability of acti-
vated sludge to efficiently remove emulsified lipids
even at very, high lipid Ioadings.
"L The activated sludge process has been shown to
exhibit poor effluent quality as Lipid:M was raised
beyond 0.25 g lipid d a y - l g - t mixed liquor sus-
pended solids. Consideration of scale up and transient
loading factors suggest that a conservative value of
O.10 g d a y - t g - t would ensure reasonable protection
against lipid overloading of the activated sludge pro-
tesS.
STEVE E. HRUDEY
4cknowledgements-.-Thls research was supported b't ,t
grant-in-aid of research provided by the Municipal Engin-
eering Branch. Pollution Control Division. Alberta
Department of Environment. Scientific advice and critical
review of this research was capably provided by Dr Roger
Perry of Imperial College.
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340
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APPENDIX I
Standard method number description or comments
Parameter
Total organic carbon 505
Total suspended solids 208D
Biochemical oxygen demand 507
Mixed liquor oxygen consumption rate 213B
Sludge volume index 213E
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(APHA, 1975}
Combustion/infrared procedure
Filtered and unfiltered samples with filtration
through Whatman G F ' C
previously fired at 500:C
Glass fibre disc (Whatman GF/C) filtration
Drying at 105~C
Dissolved oxygen measurements with YS1 Model
ARC D . O Meter and model 5720 self stirring polar-
ographic probe
l=quipment as above
Calculation based on MLSS rather than MLVSS
As per method