Pergamon

Inrcrnationul Journal.for Parasitology, Vol. 21. No. 7, pp. X2.5 xl I. 1997

0. 1997 Australian Society for Parasitology. Published by Elsevm Smnce Ltd
Printed in Great Rntam
PII: SOO20-7519(97)00037-4
0020 7519197 117~,t1~~000
and Larval. He s
Ahnasal Secretin
H. V. SIMPSON,*$ D. E. B. LAWTON,* D. C. SIMCOCK,* G. W. REYNOLDS*
and W. E. POMROYt
*Department of Physiology and Anatomy, Massey University, Palmerston North, New Zealand
TDepartment of Veterinary Pathology and Public Health, Massey University, Palmerston North,
New Zealand
(Received 9 December 1996; accepted 26 February 19971
Abatract43hp-s~ H. V., Lswton D. E. B., Sirncock D. C., Reynolds G. W. i?z Polpvoy W. E. 1997. Ef&cts
Parasitology. PgbMed by Elsevier Science Ltd.
Key words: sheep; Haemonchus contortus; experimental infection; adult worm transfer; abomasal pH; serum
gas∈ serum pepsinogen.
INTRODUCTION (Holmes & MacLean, 1971; McLeay et al., 1973; And-
Parasitism of the sheep abomasum by Haemonchus
erson et al., 1976a, 1976b, 1981, 1985; Coop et al.,
contortus inhibits gastric acid secretion and increases
1977) and in cattle by Ostertagia ostertugi (Jennings
serum pepsinogen and gastrin concentrations
et al., 1966; Fox et (II., 1987, 1993; Hilderson PI al.,
(Christie et al., 1967; Christie, 1970; Mapes & Coop, 1991)
1970, 1973; Dakkak et al., 1981, 1982; Nicholls et al.,
The adult stages of the parasite appear to be largely
1985, 1987, 1988; Blanchard & Wescott, 1985; Fox et
responsible for the altered secretory activity of the
al., 1988). These effects on gastric function are pro-
abomasum, since the transfer into cattle of adult 0.
duced also in sheep by Ostertagia circumcincta
ostertagi (McKellar et al., 1986, 1987) and 0. cir-
cumcincta into sheep (Anderson et al., 1985; Lawton
et aI., 1996) rapidly induced these ef%cts, whereas
fTo whom correspondence should be addressed. Tel: 64-
after larval infection of sheep there was a delay in
06-3.56-9099; Fax: 64-06-350-5674; E-mail: H.V.Simpson@
response until the parasite had progressed beyond
massey.ac.nz. the early larval stages (Lawton et al., 1996). Parasite
825
826 H. V. Simpson CI al
excretory/secretory (ES) products may initiate the
physiological changes in the host abomasum, since
the mucosal damage associated with the progression
of larvae to adult stages is not essential (McLeay et
al., 1973; McKellar et ul., 1987).
Evidence on the nature of the host-parasite inter-
action remains circumstantial and relies heavily on the
inter-relationships between temporal changes in the
different parameters. After infection of sheep and cat-
tle with Ostertagia, serum pepsinogen concentration
increases before either serum gastrin or abomasal pH
(Anderson et al., 1985; McKellar et al., 1986, 1987;
Lawton et al., 1996), indicating that the effects of the
parasites on abomasal secretion may not be mediated
by identical mechanisms. Increased circulating pep-
sinogen is generally attributed to a leak lesion
(Murray, 1969; Holmes & MacLean, 1971). In more
recent studies, serum gastrin and abomasal pH
initially increased together (Nicholls et al., 1988; Fox
et al., 1988; Lawton et al., 1996), suggesting that the
hypergastrinaemia may be secondary to loss of the
inhibitory effects of acid-feedback on the G cell; how-
ever, later in the parasitism, abomasal pH frequently
decreases while serum gastrin remains eIevated, so
that other factors may also be involved (Lawton et
al., 1996).
The present experiments examine the association
between increased serum pepsinogen and gastrin and
abomasal pH in sheep after infection with either larval
or adult H. contortus and also after drenching to
remove adult worms. The aim was to compare these
relationships with previous observations in sheep
infected with 0. circumcincta and identify common
features which might shed light on the mechanisms
involved.
MATERIALS AND METHODS
Experimental design. Experiment 1. Four sheep were
infected with 10000 H. contortus larvae by ruminal intu-
bation and another 4 sheep acted as uninfected controls.
Experiment 2. Four sheep were infected with approximately
9000 adult Haemonchus worms via an abomasal cannula.
After 96 h, these 4 sheep (but not control animals) were
treated orally with 0.4mg kg- ivermectin (Ivomec; Merck
& Co. Inc., Rahway, U.S.A.) and sampling was continued
for a further 192 h. Four sheep acted as uninfected controls
from which the normal range for the parameters was estab-
lished.
Animals. All animals were nematode-naive, 20-week-old
Romney-cross sheep (2 male, 2 female in each group) of body
weight 24-37 kg. A cannula was surgically inserted under
general anaesthesia into the greater curvature of the abo-
masum at the junction of the body and antral regions and
exteriorized through the ventral right flank (Hecker, 1974).
Prophylactic antibiotic treatment (Streptopen; Pitman-
Moore, 5 ml i.m. daily) was given for 3 days post-operatively.
A recovery period of 7days was allowed before exper-
imentation. The sheep were housed in individual metabolism
crates and provided with water ad lib. and fed once daily at
9.30a.m. with 200g luceme chaff plus 600g lucerne nuts.
After a control sampling period of 4 and 7 days for Experi-
ments 1 and 2, respectively. the animals were infected with
H. contortus. On a few days, some infected sheep did not
consume all the feed offered. Diarrhoea did not occur, but
Sheep 9 and 12 produced soft faeces on Day 3.
Transfer of adult H. contortus. Donor sheep were killed
by captive bolt and exsanguination 23 days after larval infec-
tion. The abomasal contents were pooled and concentrated
by sedimentation at 37C to a volume of approximately 2.5 I.
Worm counts were performed on 5 samples of 10 ml. Within
4 h of collection of the worms, the abomasa of recipient
animals were drained through the cannulae and 2~01s of
300mI of concentrated contents were infused through the
cannula at an interval of 1Smin. Each animal received
approximately 9000 23-day-old worms. No larval stages were
seen in the infusate.
Blood and abomasalfluid samples. In Experiment 1, blood
and abomasal contents were collected 30 min before, and 2-
3 h after, feeding for determination of serum pepsinogen and
gastrin and abomasal pH. After infection, sheep in Experi-
ment 2 were sampled 2 hourly for 48 h, then 4 hourly until
anthelmintic treatment. Thereafter samples were taken 4
6 hourly for the first day and then twice daily to 192 h. Blood
was collected by jugular venipuncture into plain evacuated
tubes, the serum separated and stored at -20C for gastrin
and pepsinogen assay. About 1 ml of abomasal contents was
aspirated into a syringe through the cannula and the pH
measured with a PHM82 Standard pH Meter (Radiometer,
Copenhagen).
Serum pepsinogen. Serum pepsinogen concentration was
determined in duplicate using the method of Pomroy &Char-
leston (1989). Briefly, 0.25ml of serum was acidified with
1.25ml of 0.06M HCl and incubated for 3 h at 37C. One
millilitre of 10% TCA was added, the mixture centrifuged at
3500 r.p.m. for 15 min and 1 ml of supematant added to 2 ml
of 0.5 M NaOH. The liberated tyrosine was estimated by the
addition of 0.5 ml Folin-CiocaIteau reagent and reading the
absorbance at 700 run. Pepsinogen activity was determined
from the difference in free tyrosine between the incubated
sample and a non-incubated blank and expressed as
mU tyrosine 1-l.
Serum gastrin. Serum gastrin was determined in triplicate
by a radioimmunoassay (Simpson ef al., 1993) based on the
method of Hansky & Cain (1969). The antiserum used was
Hanskys Ab74 (the generous gift of Dr J. Hansky). Synthetic
human nsG17 (Research Plus, Bayanne, NJ, U.S.A.) was
used to prepare radioactive label and standards.
Parusifology. Infective larvae (L3) were obtained from cul-
tures of faeces from sheep infected with a pure strain of H.
contortus. Larvae were confirmed to be motile. Larvae were
exsheathed with 0.2% sodium hypochlorite and rinsed with
water I h before intraruminal infection. Faecal egg counts
per g (e.p.g.) were determined using a modified McMaster
method (Stafford et al., 1994) on faecal samples collectedper
rectum. For all sheep, samples were collected before and
2 days after surgery. In Experiment 1, sheep were sampled at
weekly intervals and in Experiment 2 they were sampled
4 days after transfer of worms and 4 days later, after treat-
ment with anthelmintic.
Definition of normal values. All values for serum pepsin-
ogen, serum gastrin and abomasal pH in samples collected
Adult and larval H. contorrus infection $27
Table I-Pre-infection abomasal pH, serum gastrin and serum pepsinogen values for each infected sheep (n = 7 for Sheep
14; n = 14 for Sheep 9-12). Also shown are values from a population of parasite-naive sheep (n = 1000) used to define the
upper limit of the normal range as mean + 2 SD.
Sheep
I
2
3
4
9
IO
11
12
Population
Abomasal pH Serum gastrin (PM) Serum pepsinogeu (mU tyrosiue I !)
Mean Mean+2S.D. Mean Meanf2S.D. Mean Meani2S.D
2.61 3.33 53 15 113 xi5
2.88 3.20 56 71 62 lb2
3.04 3.41 68 92 301 531
2.97 3.30 74 95 105 451
2.68 3.22 66 84 123 255
2.74 3.34 46 62 209 317
2.65 3.31 73 125 206 318
2.71 3.29 48 74 42 IO6
2.80 3.26 38 64 223 4%
from parasite-naive sheep (controls and pre-infection) were
compared with normal population reference ranges derived
from over 1000 values for each parameter in parasite-naive
sheep (Lawton et al., 1996). The upper limits of the normal
population reference ranges (mean + 2 S.D.) were abomasal
pH, 3.26; serum pepsinogen, 454mU I- and serum gastrin,
64pM (Table 1). For each sheen, the pre-infection ranges
were calculated for each parameter and also used to define
normal values. A parameter was considered to be elevated
when 2 successive values were above the designated upper
limit.
RESULTS
Controls
There were small fluctuations in abomasal pH and
serum gastrin and pepsinogen concentration in con-
trol sheep, with the only consistent trend being an
increase in serum gas&in after feeding in some
animals. In the pre-infection period, a few sheep (par-
ticularly Sheep 3 and 11) had elevated serum gastrin
levels in anticipation of feeding which raised their
individual calculated upper limits for normal values.
Individual pre-infection values (Table 1) were gen-
erally similar to estimated population values (Lawton
et al.. 1996), although serum pepsinogen was lower.
Individual ranges were used for comparison after
infection, except for serum gastrin in Sheep 3 and 11
in which the individual range was unusually high since
the sheep sometimes became excited around feeding
time. In these animals, the population range was used
for evaluation of serum gastrin.
Injbction with larval H. contortus
In general, all 3 parameters were raised by the para-
sitism althongh the magnitude of the changes varied
between the 4 individual sheep (Fig. 1). Changes in
abomasal pH were small. The timing of changes in
abomasal pH and serum gastrin and pqtinogen was
more easily discerned when the efkts were huger, but
no consistent differences between sheep were appar-
ent. There were no increases before Day 2 and ail 3
parameters had begun to increase by Day 4 or 5. In
general, peak values were reached between Days 8 and
10, after which all decreased steadily. Abomasat pW
was usually within the normal range again by Day
15, whereas serum gas&in and pepsinogen, although
declining, generally remained elevated on Days 20.~
25, except when there had only been a small initial rise.
Sheep 2 and 4 had barely detectable serum pepsinogen
levels before and after infection and were classed as
non-responders.
Transjizr ofadult H. contortus
Although variable in magnitude between sheep,
there were increases in all parameters in all sheep
except serum pepsinogen in Sheep 12 which was a
non-responder (Fig. 2). Abomasal pH and serum
gas&in increased at about the same time (at about
38 h in Sheep 11, but at 22 h in Sheep 9 and 10 and
probably in Sheep 12 in which a small increase, but
still within the pm-infection range, occurred with the
increase in serum gastrin at 22 h). These increases were
before, or at the same time as, increases in serum
pepsinogen (serum pepsinogen was raised at 2hh in
Sheep 10 and 36-38 h. in Sheep 9 and 11).
The magnitude of the increases in abomasal pW.
serum pepsinogen and serum gas&in was independent
of one another in individual animals. Serum gastrin
continued to increase until drenching, reaching values
around 6OOpM in all 4 sheep whereas abomasai pH
reached a peak value at around 48 h. In the 3
responder sheep, the maximum value for serum pep-
828
H. V. Simpson et al.
SHEEP 1
1 1 SHEEP2 SHEEP 3
1
SHEEP 4
Fig. 1. Abomasal pH and serum gastrin and pepsinogen concentrations before (m) and 3 h after (A) feeding in 4 parasite-
naive sheep infected intraruminally with 10 000 H. contortus larvae. The horizontal dotted lines represent 2 S.D.s above the
mean values determined for each animal before infection and the solid line the corresponding population value.
1 1 SHEEP10 SHEEP 11 SHEEP 12
Fig. 2. Abomasal pH and serum gas&in and pepsinogen concentrations in 4 parasite-naive sheep into which approximately
9000 adult H. contortus were transferred through abomasal cannulae. The horizontal dotted lines represent 2 S.D.s above
the mean values determined for each animal before infection and the solid line the corresponding population value.
sinogen just before drenching represented a 4-fold within 2days and particularly rapid in Sheep i0
increase. (within 8 h). Serum gastrin and pepsinogen took in
After drenching, the time for the 3 parameters to
excess of 4 days in 2 sheep. In Sheep 10, serum gastrin
return to pre-infection levels varied. Abomasal pH
was elevated for 34days after abomasal pH was in
showed the steepest fall, being in the normal range the normal range.
Adult and larval H. contortus infection xJ.9
Parasitology
No eggs were seen in the faeces of uninfected sheep
during the experimental period. Sheep 1-4 (infected
with larvae) had no eggs in the faeces on Day 18, but
on Day 25 the faecaf egg counts (FEC) were 3050,
50, 15 850 and 17 500 e.p.g., respectively. Sheep which
received adult worms all had evidence of estab-
lishment: on Day 4 the faecal egg counts were 2100,
4100, 4150 and 2150e.p.g. for Sheep 9-12, respec-
tively. No eggs were detected after treatment with
anthelmintic.
.
DISCUSSION
This study has shown for the first time that the
direct transfer of adult Haemonchus contortus worms
into recipient sheep rapidly alters abomasal secretory
activity in a similar manner to that demonstrated in
previous studies with Ostertagia spp. in sheep and
cattle, and that the pathophysiological effects of all 3
species are linked to the presence of adult worms and
not to larval development in the mucosa. The rapidity
of recovery after worm removal by drenching high-
lights the reversibility of acid suppression in the pres-
ence of adult worms.
Adult H. contortus, but not early larval stages,
inhibited gastric acid production and raised serum
gastrin and pepsinogen concentrations, as also
accompanies the parasitism of sheep by 0. cir-
cumcincta (Anderson et al., 1985; Lawton et al., 1996)
and cattle by 0. ostertagi (McKellar et al., 1986,1987).
The initial disturbance to abomasal secretion by all of
these species of worms is unlikely to be caused by the
tissue damage associated with the histotrophic stages
of these parasites and chemical mediation would
appear more likely.
The magnitude of the responses to the different
parasites and in individual animals can be very vari-
able and was not related to the faecal egg count. In
Experiment 1, Sheep 2 and 4 were non-responders
for serum pepsinogen but had low and high FEC,
respectively, while Sheep 4 had overall the smallest
response for all 3 parameters but the highest FEC.
Infection with H. contortus larvae inhibited abomasal
secretion about 2days earlier than with 0. circum-
cincta in sheep of the same age and breed (Lawton et
ul., 1996), as might be expected from a more rapid
progression through the larval stages; effects of the
parasites on abomasal function were apparent by 4-
5 days when the 3rd moult would occur (Stoll, 1943;
Sommerville, 1954, 1963; Christie et al., 1967).
Although the responses to larval Haemonchus were
small (Fig. l), the transfer of 9000 adult worms pro-
duced very marked effects on abomasal secretion (Fig.
2), suggesting that the 2 nematode species share the
same mode of action on obomasal secretion.
The transfer of adult Haemonchus caused rapid and
more uniform effects than after larval administration
and their timing was similar to those previously seen
after adult 0. c&wncincta transfer (Lawton et al.,
1996). The principal difference was the later rise in
serum pepsinogen with H. contortus compared with
the early increase (by 2 h in some sheep) with 0. rir-
cumcincta into sheep (Anderson et al., 1985; Lawton
et al, 1996) and 0. cwtertagi into calves (McKehar et
al., 1986, 1987). No reason for this is immediately
apparent, particularly since H. contortus preferentially
establish in the fundic region, where most pe#inogen
is produced, and not in the pyloric area (Sommerville,
1963; Charleston, 1965; Dash, 1985; Rahman &
Collins, 1990). The hyperpepsinogenaemia is likely to
be caused by a different mechanism from those which
increase abomasal pH and serum gastrin. This is indi-
cated by the independence of the rise in serum pep-
sinogen from those in the other 2 parameters, as well
as the occurrence of non-responders which made
up 15-25% of the total number of sheen infected with
H. contortus in this study or with 0. circumcincra by
Lawton et al. (1996). The feature these sheep had in
common was a very low pre-infection serum pep-
sinogen concentration, which may indicate an
inherently lower leakage of pepsinogen into the cir-
culation.
Although serum gastrin initially increased at the
same time as abomasal pH and may be the result of
the withdrawal of acid feedback, the 2 parameters
were largely independent and therefore other factors
such as inflammatory mediators may be involved, at
least after the early post-infection period. The acid-
secreting parietal cell may be the principal target of
the worms, since both Ostertagia (H. V. Simpson.
unpublished data) and Haemonchus (E. Haag, 1995.
The effect of Haemonchus contortus excretory/
secretory products on abomasal secretion. Dr Med.
Vet. Thesis, Hannover) have a reduced life span in
vitro at low pH compared with their viability above
pH4.5. The ability to inhibit parietal cells may bc
essential for survival, particularly since Haemonchus
preferentially establish in the fundic region.
The mechanism of acid inhibition by abomasal
parasites, which clearly followed infection with either
adult Haemonchus or Ostertagia, is as yet unknown.
The rapidity of the onset of raised abomasal pH after
worm transfer and the rapid return to pre-infection
levels after drenching (Fig. 2) strongly support a
reversible chemical inhibition rather than the loss of
parietal cells through physical damage. The recovery
of parietal cell function began within 2 h of drenching,
too fast to be dependent on the production of new
830 H. V. Sin
cells, and was complete after about 4days. Previous
reports were similar: abomasal pH was normal within
54-172 h after drenching to remove 0. circumcincta
(Anderson et al., 1976a) or 24 h for H. contortus infec-
tion (Dakkak et al., 1985). The pH of the abomasal
contents may not accurately reflect the acid-inhibiting
activity of the worms, as a low worm burden may be
able to alter the activity of nearby cells and thus reduce
their exposure to local acid conditions without affect-
ing the overall pH of the contents. Raising the abom-
asal pH to 5 or 6 represents a significant feat on the
part of the worms.
In summary, this study has shown the similarity
between Haemonchus and Ostertagia in modifying
abomasal secretory function which is likely to be
evoked through the same mechanisms. The rapidity
of the responses to the transfer of adult worms and to
their removal by treating with anthelmintic supports
a role for worm ES products which do not appear to
be produced by the early larval stages. The similarity
of responses to H. contortus and 0. circumcinta infec-
tion suggests the involvement of the same or very
similar ES products. Identification of any chemical
mediators will probably depend on in vitro studies
using simpler systems rather than whole animal
studies.
Acknowledgements-The technical assistance of B. Adling-
ton, D. Anthony, S. Calder, B. Guthrie and B. Parlane is
acknowledged. The gas&in antibody Ab74 was the kind gift
of Dr J. Hansky. We are grateful for the generous financial
support for this work by the C. Alma Baker Trust and the
E. & C. Thorns Bequest.
REFERENCES
Anderson N., Blake R. & Titchen D. A. 1976a. Effects of a
series of infections of Ostertagia circumcincta on gastric
secretion of the sheep. Parasitology 72: l-12. -
Anderson N.. Hanskv J. & Titchen D. A. 1976b. Hvner-
gastrinaemia during a parasitic gastritis in sheep. Jo&al
of Physiology 256: 5 1-52P.
Anderson N., Hansky J. & Titchen D. A. 1981. Effects of
Ostertagia circumcincta infections on plasma gastrin in
sheep. Parasitology 82: 401-410.
Anderson N., Hansky J. L Titchen D. A. 1985. Effects on
plasma pepsinogen, gastrin and pancreatic polypeptide of
Ostertagia spp. transferred directly into the abomasum of
sheep. International Journalfor Parasitology 15: 159-165.
Blanchard J. L. & Wescott R. B. 1985. Enhancement of
resistance of lambs to Haemonchus contortus by previous
infection with Ostertagia circumcincta. American Journal
of Veterinary Research 46: 21362140.
Charleston W. A. G. 1965. Pathogenesis of experimental
haemonchosis in sheep, with special reference to the devel-
opment of resistance. Journal of Comparative Pathology
75: 55-67.
ipson et al.
Christie M. G. 1970. The fate of very large doses of Haemon-
thus contortus in the ovine abomasum. Journal of Com-
parative Pathology 80: 89-100.
Christie M. G., Brambell M. R. & Mapes C. J. 1967. Effect
of young Haemonchus contortus on abomasal pH in sheep.
The Veterinary Record 80: 207-208.
Coop R. L., Sykes A. R. & Angus K. W. 1977. The effect of
a daily intake of Ostertagia circumcincta larvae on body
weight, food intake and concentration of serum con-
stituents in sheep. Research in Veterinary Science 23: 76
83.
Dakkak A., Bueno L. & Fioramonti J. 1981. Effects of two
consecutive experimental Haemonchus contortus infections
on abomasal pepsin and electrolytes and serumpepsinogen
and electrolytes of sheep. Annals Recherche Veterinaire 12:
65570.
Dakkak A., Fioramonti J. & Bueno L. 1982. Haemonchus
contortus: abomasal transmural potential difference and
permeability changes associated with experimental infec-
tion in sheep. Experimental Parasitology 53: 209-216.
Dakkak A., Daoudi A. & Ruckebusch Y. 1985. Haemonchus
contortus and Ostertagia circumcincta: fenbendazole treat-
ment and abomasal permeability changes in sheep. Amer-
ican Journal of Veterinary Research 46: 209-211.
Dash K. M. 1985. Distribution of trichostrongylid nema-
todes in the abomasum of sheep. International Journalfor
Parasitology 15: 5055510.
Fox M. T., Carroll A. P., Hughes S. A., Uche U. E., Jacobs
D. E. & Vaillant C. 1993. Gas&in and gastrin-related
responses to infection with Ostertagia ostertagi in the calf.
Research in Veterinary Science 54: 38439 1.
Fox M. T., Gerrelli D., Pitt S. R., Jacobs D. E., Hart I. C.
& Simmonds A. D. 1987. Endocrine effects of a single
infection with Ostertagia ostertagi in the calf. International
Journalfor Parasitology 17: 1181-l 185.
Fox M. T., Pitt S. R., Gerrelli D., Jacobs D. E., Adhikari D.
R. & Goddard P. J. 1988. Use of blood gastrin assay
in the diagnosis of ovine haemonchiasis. The Veterinary
Record 122: 136-137.
Hansky J. &Cain M. D. 1969. Radioimmunoassay of gastrin
in human serum. Lancer 2: 1388-1390.
Hecker J. 1974. Experimental Surgery on Small Ruminants,
Butterworth, London.
Hilderson H., Domy P., Berghen P., Vercruysse J., Fransen
J. & Braem L. 1991. Gastrin and pepsinogen changes
during an Ostertagia ostertagi challenge infection in calves.
Journal of Veterinary Medicine B 38: 25-32.
Holmes P. H. & MacLean J. M. 1971. The pathophysiology
of ovine ostertagiasis: a study of the changes in plasma
protein metabolism following single infections. Research
in Veterinary Science 12: 265-27 1.
Jennings F. W., Armour J., Lawson D. D. & Roberts R.
1966. Experimental Ostertagia ostertagi infections in
calves: studies with abomasal cannulas. American Journal
of Veterinary Research 27: 1249.
Lawton D. E. B., Reynolds G. W., Hodgkinson S. M., Pom-
roy W. E. & Simpson H. V. 1996. Infection of sheep
with adult and larval Ostertagia circumcincta: effects on
abomasal pH and serum gastrin and pepsinogen. Znter-
national Journal for Parasitology 26: 106>1074.
Mapes C. J. &Coop R. L. 1970. The interaction of infections
of Haemonchus contortus and Nematodirus battus in lambs.
Journal of Comparative Pathology 80: 123-136.
Mapes C. J. &Coop R. L. 1973. On the relationship between
abomasal electrolytes and some population parameters of
the nematodes Haemonchus contortus and Nematodirus
battus. Parasitology 66: 95-100.
Adult and larval H. conforrus infection 3.2 I
McKellar Q., Duncan J. L., Armour J. & McWilliam P.
1986. Response to transplanted adult Usfertagia ostertagi
in calves. Research in Veterinary Science 40: 367-371.
McKellar Q. A., Duncan J. L., Armour J., Lindsay F. E. F.
& McWilliam P. 1987. Further studies on the response to
transplanted adult Ostertagia ostertagi in calves. Research
in Veterinary Science 42: 29-34.
McLeay L. M., Anderson N., Bingley J. B. & Titchen D. A.
1973. Effects on abomasal function of Ostertagia
circumcincta infections in sheep. Parasitology 66: 241-
257.
Murray M. 1969. Structural changes in bovine ostertagiasis
associated with increased permeability of the bowel wall
to macromolecules. Castroenterology 56: 763-772.
Nicholls C. D., Hayes P. R. & Lee D. L. 1987. Physiological
and microbiological changes in the abomasum of sheep
infected with large doses of Haemonchus contortus. Journal
qfComparati~e Pathology 97: 299-308.
Nicholls C. D., Lee D. L., Adrian T. E., Bloom S. R. & Care
A. D. 1985. Endocrine effects of Haemonchus contortus
infection in lambs. Regulatory Peptides 13: 85.
Nicholls C. D., Lee D. L., Adrian T. E., Bloom S. R. & Care
A. D. 1988. Hypergastrinaemia of sheep infected with
Haemonchus contortus. Research in Veterinary S&me 45:
124-126.
Pomroy W. E.&Charleston W. A. G. 1989. Failure of young
goats to acquire resistance to Haemonchus contortu7 NUL
Zealand Veterinary Journal 37: 23-26.
Rahman W. A. & Collins G. H. 1990. The establishment and
development of Haemonchus contortusin goats. Veterinar>~
Parasitology 35: 189- 193.
Simpson H. V., Reynolds G. W. & Carr D. H. 1993. Low
tissue gastrin content in the ovine distal duodenum is
associated with increased percentage of 034. Comptlrutil,e
Biochemistry and Physiology 104Az 461448.
Sommerville R. I. 1954. The histotrophic phase of the nema-
tode parasite, Ostertagia circumcincta. Australiun kwnal
of Agricultural Research 5: 130-140.
Sommerville R. I. 1963. Differential growth of Ostertagiu
spp. m the sheeps abomasum. The Journal ofPara.~i!nlo~~~~
49: 698-699.
Stafford K. J., West D. M, & Pomroy W. E. 1994. Nematode
worm egg output by ewes. New Zealand Vetrrinar), Jourrrul
42: 30-32.
Stoll N. R. 1943. The wandering of Haemonchus contorr~.~
in the sheep. Journal of Parasitologv 29: 407-416.