316 Opinion TRENDS in Parasitology Vol.17 No.

7 July 2001

Phospholipid must obtain them from the dietary products of the

host small intestine2.
In vitro, Giardia is cultured in Diamond’s medium

remodeling/generation (TYI-S-33), supplemented with bovine serum and

bile3. Bovine serum Cohn fraction IV-1 (a fraction
enriched with α-globulins, insulin-like growth factors

in Giardia: the role of and lipoproteins) has been shown to support the
growth of the parasite4. Trophozoites have also been
cultured in serum-free medium containing either an

the Lands cycle undefined lipoprotein–cholesterol–albumin solution5

Siddhartha Das, Cynthia Castillo and
Tamara Stevens
Recent results suggest that Giardia is able to carry out deacylation/reacylation
reactions (the Lands cycle) to generate new phospholipids, effectively
bypassing the de novo synthesis of the entire phospholipid molecule. The
successful operation of this deacylation/reacylation cycle is important for
Giardia because this protozoan parasite possesses limited lipid synthesis
ability. This article discusses how Giardia might use the Lands cycle to alter
phospholipids acquired from the host during its colonization in the human
small intestine.
The intestinal protozoan Giardia lamblia (also
known as Giardia duodenalis and Giardia
intestinalis) is a major cause of water-borne intestinal
infection or giardiasis in humans worldwide,
especially in developing countries1. Without proper
treatment, giardiasis can last for weeks, months or
even years. This intestinal parasite exists in two
morphological forms: trophozoites and cysts. Water-
resistant cyst forms are responsible for the
transmission of giardiasis via sewage-contaminated
drinking water, whereas trophozoites colonize the
small intestine and cause the infection. In the host
small intestine, Giardia trophozoites are exposed to
lipids and fatty acids from the host’s diet, and it is
likely that these intestinal lipids play an important
role in its life cycle. It has been proposed that Giardia
trophozoites are unable to synthesize phospholipids,
fatty acids and cholesterol de novo and, therefore,
or a mixture of biliary lipids6, suggesting that lipids
are required for giardial growth.
The uptake, intracellular localization and
metabolism of exogenous lipids has been
demonstrated using fluorescent conjugated and
radiolabeled glycerophospholipid (GPL) and fatty
acid analogs7. Analysis of cultured trophozoites
exposed to fluorescently conjugated analogs revealed
the presence of phosphatidylethanolamine (PtdEtn)
in the inner leaflet of the plasma membrane as well as
at intracellular locations, phosphatidylcholine
(PtdCho) in the plasma and flagellar membranes, and
phosphatidylglycerol (PtdGly) mostly in the
perinuclear membrane (Fig. 1), which has recently
been identified as the endoplasmic reticulum of
Giardia8. Studies with radiolabeled lipids and fatty
acids indicated that Giardia was able to transport
these molecules and incorporate them into cellular
lipids7,9,10 and proteins11–13. Lipids and fatty acids are
also important for the differentiation of this parasite,
as exposure of cultured Giardia trophozoites to
conjugated bile and fatty acids at a slightly alkaline
pH (7.8) triggers encystation14. Cholesterol
deprivation15 was also shown, in vitro, to stimulate
encystation. On the basis of these reports, it can be
concluded that exogenous lipids, fatty acids and
sterols are used by this parasite as precursors for the
synthesis of membrane and cellular lipids.
The Lands cycle and its enzymes
The rapid deacylation/reacylation of cellular
phospholipids is known as the Lands cycle. In this
cycle, cellular phospholipases generate
lysophospholipids (LPLs) and free fatty acids, which
are then recycled to produce new or altered patterns

Siddhartha Das*
Cynthia Castillo
Dept Biological Sciences,
University of Texas at
El Paso, El Paso,
TX 79968-0519, USA.
*e-mail: sdas@utep.edu.

Tamara Stevens
Dept Pathology, Gila Fig. 1. Confocal microscopy showing characteristic localization of various phospholipids (glycerophospholipids) in Giardia trophozoites. (a) BODIPY-
Regional Medical Center, phosphatidylcholine is localized in the plasma membrane (arrow) and flagellar membranes (broken arrow). (b) BODIPY-phosphatidylethanolamine is
Silver City, NM 88061, localized in the inner leaflet of the trophozoite’s plasma membrane; some is also accumulated intracellularly. (c) A substantial amount of
USA. NBD-phosphatidylglycerol is concentrated in the perinuclear membranes (arrowheads) of Giardia. Scale bars = 5.0 µm.

http://parasites.trends.com 1471-4922/01/$ – see front matter © 2001 Elsevier Science Ltd. All rights reserved. PII: S1471-4922(01)01901-8
 Opinion TRENDS in Parasitology Vol.17 No.7 July 2001 317

not only required for phospholipid remodeling, but

O are also involved in the synthesis and degradation of
R2 C O –
CH2 O CO R1 CH2 O CO R1 CH2 O CO R1
O Arachidonoyl-
R2 C O C H
CoA
Arachidonoyl O C H
HO C H
PLA2 AT
O O O
CH2 O P O X CH2 O P O X
CoA CH2 O P O X
O– O– O–
Glycerophospholipid Lysoglycerophospholipid Newly synthesized
glycerophospholipids
with arachidonic acid
substituted at sn2 position
1-Acyl-lysophosphoglyceride PLA2
Oxidation
and
cyclization
Prostaglandins Arachidonic acid
TRENDS in Parasitology
Fig. 2. The Lands cycle. Incorporation and release of a fatty acid takes place via acyltransferase and
phospholipase A2 enzymes. The fatty acid incorporated at the hydroxyl group on C-1 of the glycerol
(sn1 position) usually has a fully saturated chain, whereas the fatty acid attached at C-2 (sn2 position)
is mono- or polyunsaturated. The released fatty acid (e.g. arachidonic acid) serves as a precursor of
eicosanoid or prostaglandin biosynthesis. Abbreviations: AT, acyltransferase; CoA, Coenzyme A;
PLA2, phospholipase A2; R1-COOH, saturated fatty acid; R2-COOH, unsaturated fatty acid.
of phospholipid and fatty acids. The proper regulation
of this cycle is important to control the accumulation
of potentially toxic LPLs and fatty acids.
Since its conceptual inception more than 40 years
ago, the Lands cycle16 has received wide attention
among researchers who are interested in
understanding the mechanisms of phospholipid
remodeling and/or the functional significance of
phospholipases (PLA1 and PLA2) and
acyltransferases (ATs) in various cells and tissues17,18.
The Lands cycle also constitutes the major route for
incorporation and release of free arachidonic acid
(AA, a C20:4 unsaturated fatty acid) into cellular
phospholipids, a process that is dependent on
intracellular PLA2 activities19. For example, the
stimulation of human neutrophils by a Ca2+-
ionophore triggers the synthesis of platelet-activating
factor (PAF) and also releases free AA17. Likewise,
lung tissue synthesizes dipalmitoyl-PtdCho (a
surfactant) through this pathway, accomplished by
deacylating PtdCho at the sn2 position and
substituting a palmitic acid (PamA; C16:0) moiety at
this site20. Therefore, the deacylation/reacylation
cycle of phospholipids in mammalian cells provides an
important route to generate new phospholipids by
altering the fatty acid substituents at both the sn1
and sn2 positions of GPLs. In many cases, the fatty
acid at the sn2 position is replaced by AA, which is
stored there until it is required for eicosanoid or
prostaglandin biosynthesis (Fig. 2).
ATs, transacylases (TAs) and PLA2s are considered
key enzymes of the Lands cycle. These enzymes are
bioactive lipids, such as PAF,
N-arachidonylethanolamine (anandamide), long-
chain fatty acyl-CoA and LPLs in mammalian
tissues21. It is likely that ATs are specific and involved
in the synthesis of new phospholipids by substituting
saturated fatty acids at the sn1 position and
unsaturated fatty acids at the sn2 position of old
GPLs. There are also individual ATs with different
selectivities towards LPLs and acyl-CoAs21,22.
‘...exogenous lipids, fatty acids and
sterols are used by [Giardia] as
precursors for the synthesis of
membrane and cellular lipids.’
The transfer of sn2 fatty acid from a GPL to LPL
(without the liberation of free fatty acid) is known as a
transacylation reaction. Transacylation reactions are
classified by their requirement for coenzyme A (CoA)
and show strict acyl donor and acceptor specificities.
CoA has many binding determinants in its large
structure and is especially involved in transferring
acyl groups from one molecule to another. CoA-
dependent transacylation reactions show affinity
towards C20:4, C18:2 and C18:0 fatty acids. C20:4 and C18:2
fatty acids originate from the sn2 position of
phospholipids, whereas C18:0 is transferred from the
sn1 position. CoA-independent transacylation
reactions prefer the transfer of C20 and C22
polyunsaturated fatty acids23–27, and are also involved
in the acylation of ether-containing phospholipids24.
PLA2 cleaves fatty acids at the sn2 position of GPLs
to produce LPLs. Depending on their catalytic
functions and structural uniqueness, PLA2 isoforms
can be classified into three main categories: (1) the
small Ca2+-dependent secreted PLA2 (sPLA2) isoform;
(2) the 85 kDa Ca2+-dependent cytosolic PLA2 (cPLA2)
isoform; and (3) the 80 kDa Ca2+-independent
cytosolic PLA2 (iPLA2) isoform28. The inhibition of
iPLA2 (using antisense RNA and bromoenol lactone)
reduces the incorporation of AA and other fatty acids
into membrane phospholipids of P388D1
macrophages and Chinese hamster ovary cells,
suggesting that iPLA2 plays an important role in
phospholipid remodeling29. Exogenously added
human sPLA2 was shown to be involved in releasing
AA from adherent mammalian cells29. Furthermore,
two Ca2+-independent PLA2s (group VII and VIII)
have been identified and found to be specific for the
synthesis of PAF28. The mobilization of oleic acid in
lipopolysaccharide-stimulated P388D1 macrophages
is a result of groupV sPLA2 activities30. Therefore,
based on these reports, it is reasonable to postulate
that PLA2, like AT and TA, is also specific and
extremely important for the successful operation of
the Lands cycle.

http://parasites.trends.com
318 Opinion TRENDS in Parasitology Vol.17 No.7 July 2001

myristic acid (MA; C14:0) into cellular GPLs and

O
O
glycosylphosphatidylinositol molecules7,11, and we
R2 C O– have recently demonstrated that two unsaturated
R3 C O CoA
CH2 O CO R1 CH2 O CO R1 CH2 O CO R1
fatty acids – AA and oleic acid (OA; C18:1) – are
O O
R3
incorporated into GPLs by Giardia9. AA was
R2 C O C H HO C H C O C H
PLA2 AT transported more rapidly than other fatty acids tested
O O O
(e.g. PamA, MA and OA), suggesting that this C20:4
CH2 O P O X CH2 O P O X CoA CH2 O P O X
O– O– O–
unsaturated fatty acid might play an important role in
Glycerophospholipid Lysoglycerophospholipid Glycerophospholipid phospholipid metabolism. Ellis et al.31, however, failed
remodeled at sn2 position to detect lipid-bound AA in Giardia trophozoites,
suggesting that it might be present only in trace
TRENDS in Parasitology amounts. The preferential incorporation of these fatty
acids into various GPLs (i.e. PtdIns, PtdCho and
Fig. 3. Proposed pathway for the incorporation of free fatty acids into giardial phospholipids. PtdGly) is most likely catalyzed by the enzymes of the
Although the incorporation of unsaturated fatty acids (i.e. arachidonic acid) at the sn2 position is more Lands cycle as proposed in Fig. 3.
common, the incorporation of a saturated fatty acid (e.g. palmitic acid) at this position cannot be ruled
out. Abbreviations: AT, acyltransferase; CoA, Coenzyme A; PLA2, phospholipase A2; R1-COOH,
[14C]PamA-containing lysophosphatidylcholine
saturated fatty acid; R2-COOH, unsaturated fatty acid; R3-COOH, saturated or unsaturated fatty acids. (LPtdCho) might serve as a donor of sn1 fatty acid,
with giardial PtdGly or other GPLs acting as acceptor
Does Giardia use the Lands cycle to generate new molecules9 (Fig. 4a). Likewise, [3H]PamA-containing
phospholipids? PtdCho might donate its sn2 fatty acid to giardial
Like higher eukaryotes, Giardia was shown to take up GPLs (Fig. 4b). The existence of giardial
and incorporate free and conjugated (conjugated with phospholipases (PLA1 and PLA2) was demonstrated
a GPL or LPL) fatty acids into cellular by an experiment where trophozoites were incubated
phospholipids7,9,10. [3H]AA was incorporated into a with a mixture of [14C]LPtdCho and [3H]PtdCho, and
wide variety of neutral lipids and phospholipids, both [14C] and [3H] fatty acids were transferred to
whereas [3H]PamA was incorporated into various acceptor molecules9. Furthermore, aristolochic acid,
GPLs including phosphatidylinositol (PtdIns)10. A an inhibitor of Ca2+-ionophore-stimulated PLA2, was
fatty acid desaturase activity (which can convert shown to inhibit the transfer of sn2 fatty acid from
monounsaturated fatty acids to polyunsaturated fatty [3H]PtdCho to PtdGly (acceptor molecule) but not
acids) has been demonstrated in this organism31. from [14C]LPtdCho, once again implicating a Ca2+-
Initial work showed that Giardia transfers PamA and dependent PLA2 activity in transferring the sn2 fatty
acid to an acceptor (GPL or LPL) molecule9. However,
it is not known whether these transfer reactions are
(a) O
R4 C OH
CoA-dependent or -independent. Based on these
CH2 O CO R1* CH2 O CO R3 CH2 O CO R3 results it can be postulated that Giardia is capable of
O O remodeling its phospholipids by deacylation/
HO C H + R4 C O C H R1 C O C H reacylation reactions. Although the detailed steps of
AT for R1 COOH these reactions are yet to be identified, we speculate
O O O
that the enzymes of the Lands cycle are involved in
CH2 O P O X CH2 O P O X CH2 O P O X
this process.
O– O– O–
In other parasites, the Lands cycle is not only
Lysoglycerophospholipid Glycerophospholipid Remodeled
(acyl donor) (acyl acceptor) glycerophospholipid
involved in phospholipid remodeling but also plays an
important role in host–parasite interactions. Studies
(b) of the production of prostaglandins and eicosanoids in
R4 C OH
CH2 O CO R1 CH2 O CO R3 CH2 O CO R3 parasites have focused primarily on their role in
O parasite growth and penetration of host cells and
O O
*R2 C O C H + R4 C O C H R2 C O C H
tissues, as well as evasion of the host immune system.
AT for R2 COOH Trypanosoma brucei possesses a cell-associated,
O O O Ca2+-stimulated PLA2 that catalyzes the release of AA
CH2 O P O X CH2 O P O X CH2 O P O X and thus regulates the production of eicosanoids32.
O– O– O–
Similarly, Spirometra erinacei has been shown to
Glycerophospholipid Glycerophospholipid Remodeled metabolize AA to prostaglandin E2, which might
(acyl donor) (acyl acceptor) glycerophospholipid
relate to larval escape mechanisms from the host
immune system33. The rapid release, incorporation,
TRENDS in Parasitology modification and turnover (by deacylation/
reacylation) of fatty acids to PtdCho was shown to
Fig. 4. Fatty acid remodeling of phospholipids by conjugated fatty acids. In (a), lysoglycerophospholipid occur in adult schistosomes and thought to be
acts as an acyl donor (sn1 fatty acid) and in (b) glycerophospholipid donates its sn2 fatty acid. In both
involved in host membrane destabilization as well as
cases (a and b) either glycerophospholipid or lysoglycerophospholipid (not shown here) might act as
an acceptor molecule. The asterisk denotes radiolabeled (14C or 3H) palmitic acid. R3-COOH, saturated immune evasion by the parasite34. Schistosoma
fatty acid and R4-COOH, unsaturated fatty acid (speculative). mansoni and Trichobilharzia ocellata synthesized the

http://parasites.trends.com
 Opinion TRENDS in Parasitology Vol.17 No.7 July 2001 319

Acknowledgements
We thank Luis Irwin and
Steve Aley for helpful
comments on this article.
This work was supported
by National Institutes of
Health grants
5G12RR08124-08 and
S06GM08012-30.
same types of prostaglandins, leukotrienes and
hydroxyeicosatetranoic acids that inhibited
superoxide production in human neutrophils35.
Intrinsic or secreted products of Entamoeba
histolytica have been shown to cause dramatic
alterations of AA metabolism and eicosanoid levels in
macrophages of infected animals36. It remains to be
seen if Giardia, like other parasites, is capable of
synthesizing eicosanoids or prostaglandins.
Concluding remarks
The accumulated data on lipid recruitment,
modification and utilization by Giardia contribute to
our understanding of the mechanisms by which this
parasite exploits the host environment to synthesize
its membrane and cellular phospholipids. In addition
to deacylation/reacylation reactions, a recent report
suggests that new lipids can be produced by base
(headgroup) exchange reactions37, suggesting that
Giardia relies on fatty acid and headgroup exchange
reactions rather than de novo pathways2 to
synthesize new phospholipids. Investigating the
deacylation/reacylation reactions of phospholipids in
Giardia (as well as in other protozoan parasites) will
increase our knowledge regarding the possible
relationship between lipid metabolism and
pathogenesis. It is also possible that the enzymes of
the Lands cycle (i.e. ATs, TAs and PLAs) might serve
as potential targets for the development of drugs
against giardiasis.

References
1 Thomson, R.C.A. et al. (2000) Nomenclature and
genetic groupings of Giardia infecting mammals.
Parasitol. Today 16, 210–213
2 Jarroll, E.L. et al. (1981) Lipid and carbohydrate
metabolism of Giardia lamblia. Mol. Biochem.
Parasitol. 2, 187–196
3 Diamond, L.S. et al. (1978) A new medium for the
axenic cultivation of Entamoeba histolytica and
other Entamoeba. Trans. R. Soc. Trop. Med. Hyg. 27,
487–488
4 Lujan, H.D. et al (1994) Serum Cohn fraction
IV-I supports the growth of Giardia lamblia in vitro.
Infect. Immun. 62, 4664–4666
5 Reiner, D.S. et al. (1995) A lipoprotein-cholesterol-
albumin substitute stimulates Giardia lamblia
encystation vesicle formation. J. Eukaryot.
Microbiol. 42, 622–627
6 Gillin, F.D. et al. (1986) Biliary lipids support
serum-free growth of Giardia lamblia. Infect.
Immun. 53, 641–645
7 Stevens, T.L. et al. (1997) Uptake and cellular
localization of exogenous lipids by Giardia lamblia,
a primitive eukaryote. Exp. Parasitol. 86, 133–143
8 Soltys, B.J. et al. (1996) Identification of
endoplasmic reticulum in the primitive eukaryote
Giardia lamblia using cryoelectron microscopy and
antibody to BiP. J. Cell Sci. 109, 1909–1917
9 Gibson, G.R. et al. (1999) Giardia lamblia:
incorporation of free and conjugated fatty acids into
glycerol-based phospholipids. Exp. Parasitol. 92,
1–11
10 Blair, R. and Weller, P.F. (1987) Uptake and
esterification of arachidonic acid by trophozoites of
Giardia lamblia. Mol. Biochem. Parasitol. 25, 11–18
11 Das, S. et al. (1991) A surface antigen of Giardia
lamblia with a glycosylphosphatidylinositol anchor.
J. Biol. Chem. 266, 21318–21325
12 Lujan, H.D. et al. (1995) Isoprenylation of proteins
in the protozoan Giardia lamblia. Mol. Biochem.
Parasitol. 72, 121–127
13 Papanastasiou, P. et al. (1997) The variant-
surface protein of Giardia, VSP41, is a
glycosylated and palmitoylated protein.
Biochem. J. 322, 49–56
14 Gillin, F.D. et al. (1987) Encystation and expression
of cyst antigens by Giardia lamblia in vitro. Science
235, 1040–1043
15 Lujan, H.D. et al. (1996) Cholesterol starvation
induces differentiation of the intestinal parasite
Giardia lamblia. Proc. Natl. Acad. Sci. U. S. A. 93,
7628–7643
16 Lands, W.E.M. (2000) Stories about acyl chains.
Biochim. Biophys. Acta 1483, 1–16
17 Reinhold, S.L. et al. (1989) Phospholipid remodeling
in human neutrophils. J. Biol. Chem. 264,
21652–21659
18 Yamashita, A. et al. (1997) Acyltransferases and
transacylases involved in fatty acid remodeling of
phospholipids and metabolism of bioactive lipids in
mammalian cells. J. Biochem. 122, 1–16
19 Balsinde, J. et al. (1995) Inhibition of calcium-
independent phospholipase A2 prevents arachidonic
acid incorporation and phospholipid remodeling in
P388D1 macrophages. Proc. Natl. Acad. Sci.
U. S. A. 92, 8527–8531
20 Vance, D.E., ed. (1989) Phosphatidylcholine
Metabolism, CRC Press
21 Sugiura, T. et al. (1987) Transacylation of lyso
platelet-activating factor and other lyso-
phospholipids by macrophage microsomes. Distinct
donor and acceptor selectivities. J. Biol. Chem. 262,
1199–1205
22 Kerkhoff, C. et al. (1998) Substrate specificity of
acyl-CoA: lysophospholipid acyltransferase (LAT)
from pig spleen. Arch. Biochem. Biophys. 351,
220–226
23 Sugiura, T. et al. (1988) Coenzyme A-dependent
transacylation system in rabbit liver microsomes.
J. Biol. Chem. 263, 17490–17498
24 Robinson, M. et al. (1985) Acylation of
lysophospholipids by rabbit alveolar macrophages.
Specificities of CoA-dependent and CoA-
independent reactions. J. Biol. Chem. 260,
7889–7895
25 Waku, K. and Lands, W.E.M. (1968) Acyl coenzyme
A: 1-alkenyl-glycero-3-phosphorylcholine
acyltransferase action in plasmalogen biosynthesis.
J. Biol. Chem. 243, 2654–2659
26 Waku, K. and Nakazawa, Y. (1970) Acyltransferase
activity to 1-alkenyl-glycero-3-phosphorylcholine in
sarcoplasmic reticulum. J. Biochem. 68, 459–466
27 Waku, K. and Nakazawa, Y. (1977) Regulation of
the fatty acid composition of alkyl ether
phospholipid in Ehrlich ascites tumor cells. The
substrate specificities of 1-O-alkylglycerol
3-phosphate and 1-O-alkylglycerol-3-
phosphorylcholine acyltransferases. J. Biochem.
82, 1779–1784
28 Balboa, M.A. et al. (1997) Identity between the Ca2+-
independent phospholipase A2 enzymes from
P388D1 macrophages and Chinese hamster ovary
cells. J. Biol. Chem. 272, 8576–8580
29 Bezzine, S. et al. (2000) Exogenously added human
group X secreted phospholipase A2 but not the group
IB, IIA, and V enzymes efficiently release
arachidonic acid from adherent mammalian cells. J.
Biol. Chem. 275, 3179–3191
30 Balsinde, J. et al. (2000) Group V phospholipase A2-
mediated oleic acid mobilization in
lipopolysaccharide-stimulated P388D1
macrophages. J. Biol. Chem. 275, 4783–4786
31 Ellis, J.E. et al. (1996) Changes in lipid composition
during in vitro encystation and fatty acid
desaturase activity of Giardia lamblia. Mol.
Biochem. Parasitol. 81, 13–25
32 Eintracht, J. et al. (1998) Calcium entry in
Trypanosoma brucei is regulated by
phospholipase A2 and arachidonic acid.
Biochem. J. 336, 659–666
33 Fukushima, T. et al. (1993) The metabolism of
arachidonic acid to prostaglandin E2 in
plerocercoids of Spirometra erinacei. Parasitol. Res.
79, 634–638
34 Brouwers, J.F.H.M. et al. (1999) Studies on
phospholipid turnover argue against sloughing of
tegumental membranes in adult Schistosoma
mansoni. Parasitology 119, 287–294
35 Nevhutalu, P.A. et al. (1993) Schistosoma mansoni
and Trichobilharzia ocellata: comparison of secreted
cercarial eicosanoids. J. Parasitol. 79, 131–133
36 Wang, W. and Chadee, K. (1992) Entamoeba
histolytica alters arachidonic acid metabolism in
macrophages in vitro and in vivo. Immunology 76,
242–250
37 Subramanian, A.B. et al. (2000) Role of exogenous
inositol and phosphatidylinositol in
glycosylphosphatidylinositol anchor synthesis of
GP49 by Giardia lamblia. Biochim. Biophys. Acta
1483, 69–80
Erratum
In the June issue of Trends in
Parasitology, the e-mail address for
the corresponding author of the
Review article ‘A new lungworm in
muskoxen: an exploration in Arctic
parasitology’ by S.J. Kutz, E.P. Hoberg
and L. Polley [(2001) Trends Parasitol.
17, 276–280] was given incorrectly.
It should read: susan.kutz@usask.ca

http://parasites.trends.com