Gastrin and gastrin-related responses to infection with Ostertagia ostertagi in the half

The effects of a single challenge with 60,000 infective Ostertagia ostertagi larvae on blood and gastrointestinal mucosal gastrin concentrations, gastrin-producing G-cell numbers in the pyloric mucosa and growth of different parts of the gut were investigated in 16, two-and-a-half-month-old calves. Infected calves exhibited a rise in abomasal pH which was accompanied by a 145 per cent increase in wet weight of the fundic mucosa (P<0·05) and a significant rise in blood total gastrin concentrations (P<0·01). Circulating little gastrin (G-17) was unaffected. Pyloric mucosal total gastrin concentrations remained unaltered in the infected calves until day 28 when levels fell to 36·9 per cent of control group values (P<0·01). Pyloric mucosal G-cell numbers declined during the experiment in the infected group. It is suggested that release of previously stored tissue gastrin and not a change in Gcell numbers contributes to the hypergastrinaemia associated with ostertagia infection in the calf.     

Vagal and splanchnic afferent nerves are not essential for anorexia associated with abomasal parasitism in sheep

Heavy burdens of the abomasal nematode, Ostertagia (Telodorsagia) circumcincta, in growing lambs result in a reduction in liveweight gain due largely to a drop in voluntary feed intake. The present study investigated: (1) the role of subdiaphragmatic vagal and non-vagal visceral afferent nerves in mediating a reduction in voluntary feed intake, using subdiaphragmatic vagal deafferentation (vagotomy) either alone or in combination with coeliac-superior mesenteric ganglionectomy (vagotomy and sympathectomy); and (2) the association between appetite, abomasal pH, selected blood values (amidated gastrin (G-17-amide), glycine-extended gastrin (G-17-Gly), pepsinogen and leptin) and worm burden, in sheep experimentally infected with 100,000 O. circumcincta infective larvae per os. Neither vagotomy alone nor vagotomy and sympathectomy in combination adversely affected the establishment or course of development of the parasite burden, when compared with a control group subject to sham surgery. Furthermore, neither surgical procedure prevented the drop in appetite seen 5-10 days post-infection, although combined vagotomy and sympathectomy did reduce voluntary feed intake prior to the start of the study. Ostertagia infection resulted in a significant increase in abomasal pH in all three groups, which was accompanied by an increase in blood G-17-amide and in G-17-Gly, the latter reported for the first time in parasitized ruminants. There were no significant differences in blood leptin, also reported for the first time in parasitized sheep, either between groups or in comparison with pre-infection levels, though weak negative correlations were established between blood leptin and appetite from day 5 to the end of the study in all three groups and a positive correlation with blood G-17-amide in the control group over the same period. These data suggest that neither intact subdiaphragmatic vagal afferent nerves or coeliac-superior mesenteric ganglion fibres, nor changes in circulating gastrin and leptin concentrations play a major role in mediating the hypophagic effects of O. circumcincta in parasitized sheep.     

Abomasal secretion in sheep receiving adult Ostertagia circumcincta that are prevented from contact with the mucosa

AIMS: Both adult and larval Ostertagia circumcincta infections raise abomasal pH and serum gastrin and pepsinogen concentrations, either because of physical effects of the worms or from their chemical secretions. The study was designed to examine whether adult worms require contact with the gastric mucosa to effect changes in gastric secretion. METHODS: Abomasal pH and serum gastrin and pepsinogen concentrations were measured in four groups of sheep: Group A (n = 4): abomasal contents containing about 18,000 adult O. circumcincta were obtained from donor sheep, concentrated and infused into 5 microm mesh porous bags attached to surgically implanted abomasal cannulae. A second worm transplantation was carried out 1 week later to assess worm survival after 16 hours. Group B (n = 4): about 9000 adult worms, recovered after migration out of abomasal contents set in agar, were placed in small 5 microm mesh bags which were inserted through indwelling abomasal cannulae and left for 3 days. Group C (n = 2): about 3000 adult worms from the population recovered from agar were infused through abomasal cannulae which allowed. free movement in the abomasum. Group D (n = 3) was left uninfected. RESULTS: Worms transplanted directly into the abomasum (Group C) caused rapid and marked effects on abomasal secretion. Adult O. circumcincta died within 16 hours of transfer into the abomasum when they were restrained within porous bags. Nevertheless, in sheep receiving 18,000 worms, abomasal pH increased soon after new feed was presented on Days 1, 2 and 4 after worm transfer; serum gastrin was elevated in three sheep from 113 hours and serum pepsinogen increased in one animal. Sheep receiving 9000 worms showed similar trends but the results were equivocal. CONCLUSIONS: Adult O. circumcincta prevented from physical contact with the gastric mucosa by restraint in porous bags are able to raise abomasal pH. This study implicates parasite excretory-secretory products in mediating changes in gastric secretion caused by adult abomasal worms.     

Further studies on the response to transplanted adult Ostertagia ostertagi in calves

Calves infected by surgical transplantation of adult Ostertagia ostertagi had raised levels of plasma pepsinogen and those in which the largest number of worms established also had elevated plasma gastrin concentrations. Despite the elevated plasma pepsinogen values, the abomasal pH of the animals did not change significantly, and there was no significant difference in the percentage establishment of adult parasites in calves previously infected with O ostertagi third stage larvae and those which had been maintained parasite-naive before transplant.     

The relationship between appetite and plasma non-esterified fatty acid levels in housed calves

Non-esterified fatty acid (NEFA) levels in housed, 3-month-old calves were monitored in two experiments designed to establish (i) the effect of diet and time of sampling in relation to feeding on blood levels; and (ii) the relationship between appetite and plasma NEFA in animals on chopped hay. In the first experiment, a significant drop in NEFA levels was recorded after feeding. Furthermore, the NEFA levels of calves maintained on concentrate were significantly lower than those on hay throughout the post-feeding period. In the second study, a significant negative correlation was established between NEFA levels and intake of chopped hay in (i) ad-lib-fed calves infected with the abomasal nematode Ostertagia ostertagi; (ii) worm-free animals pair-fed with those in group (i); and (iii) ad-lib-fed worm-free calves. Regression analysis of raw and transformed data from all three groups combined also yielded significant correlations. It is suggested that the measurement of blood NEFA may be a useful indirect indicator of feed intake in conditioned housed calves offered hay diets.     

Gastrin and pepsinogen changes during an Ostertagia ostertagi challenge infection in calves

Daily changes in serum gastrin and pepsinogen concentration have been studied during two types of infection with Ostertagia ostertagi in calves. In a first experiment two calves were trickle infected (10 times 10,000 L3 Ostertagia) and two animals received a single infection of 100,000 L3 Ostertagia. Gastrin and pepsinogen changes are discussed in relation to adult wormburdens. The second experiment involved 8 calves and was designed to investigate pepsinogen and gastrin changes following a challenge infection in previously sensitized calves. The high dosed group was infected with 5,000 L3 O. ostertagi during 30 days, the low dosed group received 500 L3 O. ostertagi and group 3 served as uninfected control. At day 41 post infection all animals were treated with oxfendazole and on day 61 challenged with 100,000 L3 O. ostertagi. Only in the high dosed group a distinct pepsinogen and gastrin reaction was noticed. Both parameters dropped to almost preinfection levels after treatment. Two days post challenge a moderate rise (+/- 1,000 mU tyr) of the pepsinogen concentration was observed in the previously infected animals and gastrin showed a temporary slight increase in several animals 8 to 10 days post challenge. The effect of treatment and challenge infection is discussed in relation to gastrin and pepsinogen changes and immunity.     

Pathophysiology of abomasal parasitism: is the host or parasite responsible?

Nematode larvae developing within the glands cause local loss of parietal cells and mucous cell hyperplasia whereas reduced acid secretion, increased serum gastrin and pepsinogen concentrations and generalized histological changes are associated with parasites in the abomasal lumen. Parietal cells with dilated canaliculi and/or degenerative changes typical of necrosis are present soon after the transplantation of adult worms, and abomasal secretion is also affected. Anaerobic bacteria survive in greater numbers as the pH rises, with bacterial densities becoming similar to ruminal populations at an abomasal pH of 4 and above. Failure to lyse bacteria may affect adversely the nutrition of the host. The parasites may initiate the pathophysiology through the release of excretory/secretory (ES) products which either act directly on parietal cells or indirectly through enterochromaffin-like (ECL) cells by provoking inflammation or by disrupting the protective mucosal defence system.Parietal cell dysfunction is proposed as a key event which leads to loss of mature chief cells and mucous cell hyperplasia, as well as hypergastrinaemia. Inflammation increases circulating pepsinogen concentrations and may also contribute to increased gastrin secretion. Stimulation of mucosal proliferation and differentiation of parietal cells in the isthmus by the raised serum gastrin levels will be beneficial by generating a new population of active parietal cells and adequate acid secretion.     

Reduced Ostertagia circumcincta burdens in milk-fed lambs

AIMS: To compare the susceptibility to parasitism by Ostertagia circumcincta of lambs fed entirely with bovine milk or weaned on to solid feed at 3 weeks of age. In addition, the effect of a single daily feed of milk on worm burdens was assessed. METHODS: Eight lambs were assigned to each of the 3 diets: milk (M), milk plus solid feed (cereal-based pellets and lucerne chaff) (MS), or solid feed only (S). Those to be fed solid feed were converted from complete milk feeding to the designated diet during their third week of life. From 3 weeks of age, all lambs were infected with 1000 O. circumcincta larvae twice weekly for 6 weeks; 4 lambs from each diet group were given normal sheathed L3 and another 4 were infected with exsheathed larvae. Faecal egg counts (FEC) and serum gastrin and pepsinogen concentrations were monitored from Day 17 after first infection, and worm burdens and abomasal pH and morphology were determined at necropsy. RESULTS: Total worm burdens and FEC were significantly lower in the M than MS and S groups, whereas there was no significant difference between those receiving sheathed and exsheathed larvae. The milk-fed lambs had a smaller reticulo-rumen and omasum and a more acidic abomasal pH. Serum gastrin and pepsinogen were increased in all groups, irrespective of diet or type of larvae used for infection. CONCLUSIONS: The cause of the lower worm establishment in lambs fed only milk was probably not failure to exsheath in the immature gastro-intestinal tract, as there were similar worm burdens in lambs whether sheathed or exsheathed larvae were administered. The lower pH of the abomasal contents of the preruminant lambs may have been a factor, as the parasites have previously been shown to die more rapidly in vitro at low pH. Alternatively, the milk itself had adverse effects on the parasites, but was ineffective when combined with solid feed. There was no benefit from feeding a milk plus solid diet over a solid diet.     

Hypergastrinaemia of sheep infected with Haemonchus contortus

Radioimmunoassay of plasma gastrin has shown that a hypergastrinaemia occurs in yearling sheep, after infection with 10(6) infective third stage larvae of Haemonchus contortus. Plasma gastrin concentrations began to rise two to four days after infection, that is, at about the same time as, or slightly before, the abomasal pH was observed to increase.     

Blood gastrin and pepsinogen responses to subclinical infection with Ostertagia ostertagi in adult dairy cattle

Blood gastrin and pepsinogen responses to a single infection with 100,000 Ostertagia ostertagi infective larvae in lactating dairy cows were investigated. None of the infected cows showed signs of clinical ostertagiasis, nor was there any difference in live weight gain, milk yield or faecal egg count between groups. Pepsinogen levels of the infected group were significantly elevated between days 3 and 24 after infection (peak 1041 mU tyrosine; day 14). In contrast, there was no significant difference in blood gastrin levels between infected and control animals suggesting that few adult worms had become established in the former group. These data are compared with the increases in both gastrin and pepsinogen levels recorded in susceptible calves exposed to the same level, pattern and strain of ostertagia infection in a previous experiment. It is suggested that gastrin assay may be of value in adult cattle for indicating when elevated pepsinogen levels are merely associated with a rise in larval intake and not with the establishment of large adult worm burdens.     

A sequential study of the pathology associated with the infection of sheep with adult and larval Ostertagia circumcincta

Disturbances in the physiology of the abomasa of sheep infected with either adult Ostertagia circumcincta given via abomasal cannulae, or larvae (L3) given intraruminally were matched by pathological changes in tissues collected by repeated mucosal biopsy. Within 2-3 days of the transplant of adult worms, abomasal pH had increased markedly in five out of six animals, and there also had been rapid increases in serum gastrin and pepsinogen concentrations in all animals. Reductions in parietal cell number were recorded as early as 1 day after the transplant of adults and were associated with the rapid accumulation of many neutrophils and eosinophils. Mucosal hyperplasia, with increased numbers of cells closer in appearance to mucous/mucous neck cells, was a relatively late development, being most pronounced in the latter part of the infection. In sheep given larvae, changes in secretory physiology were again matched by a concurrent fall in parietal cell number and by the accumulation of inflammatory cells. Changes became maximal when most worms could be expected to be present as adults, confirming the role of adults in the natural disease. Some abnormalities were detected in biopsies collected from animals maintained free of parasites and, although milder in degree, there were similarities to those observed in parasitised tissues, there being fewer parietal cells, a modest degree of mucous cell hyperplasia and inflammatory infiltrates of predominantly neutrophils. These changes were the likely result of trauma to the tissues in the immediate vicinity of the cannula, due either to the presence of the cannula itself or to the frequent collection of biopsy material from areas close to it.     

Effect of sodium cromoglycate on the response to Ostertagia circumcincta in adult sheep

Serum pepsinogen concentrations rose more rapidly and to higher levels in adult sheep infected with Ostertagia circumcincta and treated orally with the mast cell stabilising agent sodium cromoglycate than they did in adult sheep infected with the parasite which remained untreated. Sodium cromoglycate did not affect the serum pepsinogen concentrations or abomasal pH in uninfected sheep.     

Abomasal contents of parasitised sheep contain an inhibitor of gastrin secretion in vitro

Serum gastrin concentrations are typically elevated in parasitised sheep; however, in some animals serum gastrin concentrations may fall abruptly despite a very high abomasal pH. Although proliferating abomasal bacteria in culture generate a potent inhibitor of in vitro gastrin secretion, this inhibitor has not been detected in abomasal contents of unparasitised sheep. In sheep parasitised by O. circumcincta, all abomasal fluid samples of pH 5 and above were inhibitory to gastrin release in vitro. Inhibitory activity and abomasal pH were correlated in two separate experiments; the model best fitting the data being sigmoidal in each case, with zero activity at pH 3.6 and 4.6, respectively. There was no clear evidence that the presence of a gastrin inhibitor in the abomasal contents reduced the serum gastrin concentration in parasitised sheep. Serum gastrin was correlated with abomasal pH (log(10) serum gastrin concentrations conformed to log-linear sigmoidal models).     

Haemonchus contortus and Ostertagia circumcincta: fenbendazole treatment and abomasal permeability changes in sheep

Effects of fenbendazole (7.5 mg/kg of body weight infused during 90 minutes) on abomasal transmural potential difference (PD), pH, and ionic concentrations were determined in 8 lambs fitted with permanent abomasal cannulae. Four sheep were treated before inoculation and on day 28 after inoculation, with 200,000 3rd-stage Ostertagia circumcincta larvae and 4 others were similarly inoculated with 15,000 3rd-stage Haemonchus contortus larvae. In worm-free lambs, the unique effect was an increase in Na+ concentration of the abomasal contents. The O circumcincta or H contortus-inoculated lambs had an increase in PD, pH, and Na+ concentrations. Treatment further increased these concentrations, but the effects on PD occurred and ceased earlier in sheep inoculated with O circumcincta than in those inoculated with H contortus. The results indicated that part of the changes were linked to substances produced by the parasites or by the host. The increase in PD and pH was associated with a decrease in K+ concentration and indicated that these substances stimulated HCO3- transfer.     

Stimulated pepsinogen secretion from dispersed abomasal glands exposed to Ostertagia species secretion

Carbachol and secretions from Ostertagia species parasites significantly (P less than 0.001) stimulated isolated preparations of dispersed gastric glands from bovine and ovine abomasal mucosa to secrete pepsinogen. Atropine reduced the response to both secretagogues. Live adult and larval stages of Ostertagia ostertagi and O circumcincta and homogenates of these parasites did not significantly (P greater than 0.05) increase pepsinogen production from bovine or ovine gland preparations.     

Significance of serum pepsinogen and abomasal pH levels in a field infection of O circumcincta in lambs.

Serum pepsinogen estimations from serially bled lambs grazing on pasture from spring to autumn showed correlations with the availability of Ostertagia larvae on pasture, with faecal egg counts of O circumcincta, and with Ostertagia worm counts in similar lambs slaughtered fort-nightly from the same pasture. In the slaughtered lambs correlations were recorded between worm count, serum pepsinogen level and abomasal pH. The value of serum pepsinogen estimations as a diagnostic test is discussed with reference to these findings.     

Effect of Ostertagia circumcincta excretory/secretory products on gastrin release in vitro.

It has been suggested that parasite excretory/secretory (ES) products may be capable of direct stimulation of gastrin secretion and of contributing to the hypergastrinaemia typical of abomasal parasitism. Ostertagia circumcincta ES products were tested on an ovine antral mucosal preparation which had been developed for a pharmacological study of gastrin secretion in the sheep. Its responsiveness to chemical stimulation was established by stimulation with amino acids and amines: tryptophan (0.1-5 mM) and phenylalanine (10-100 mM) stimulated gastrin release (151-160 and 117-129%, respectively), whereas glycine (0.1-100 mM) was without effect; ammonium sulphate, but not sodium sulphate, stimulated gastrin release in concentrations from 1mM (122%) to 50mM (148%). ES products were prepared by incubation of exsheathed third-stage larvae (L3) or parasites recovered on Day 8 p.i. (L4), Day 12 p.i. (10% L4, 90% immature adults), Day 21 p.i. (5% L4, 30% immature adults, 65% adults), Day 22 p.i. (20% immature adults, 80% adults), Day 30 p.i. (adults) and Day 35 p.i. (adults), or a mixed-age parasite population. Worms were recovered from agar and incubated in either distilled water or Hank's balanced salt solution (HBSS) adjusted to pH 2.5, 3.5, 4.5, 5.0 or 7.4. HBSS pH 7.4 was also prepared with antibiotics, without glucose, and with antibiotics but without glucose. Survival of Day 21 and 35 worms and exsheathed L3 in water or in a series of HBSS adjusted to pH 2.5, 3.5, 4.5, 5.0 or 7.4 was assessed from the percentage of motile parasites. L3 slowly became immotile over several days except in HBSS pH 2.5, in which survival was reduced, whereas adult worms did not tolerate incubation at 37 degrees C in water or HBSS at pH 2.5, retained motility for about 2 days at pH 3.5, but survived well at pH 4.5 and above. Incubates prepared from all stages of O. circumcincta, both in media favourable and unfavourable for parasite survival, failed to stimulate consistently the secretion of gastrin by tissue from both parasite-naive and previously exposed sheep, whereas a considerable number of incubates were significantly inhibitory. The inhibitor may not be produced by the nematodes, but by contaminating abomasal or environmental microflora, as inhibitory activity was predominantly generated by prolonged incubation, it was less potent when glucose was omitted and was not present in media containing antibiotics. This study did not find evidence for a gastrin stimulant in O. circumcincta ES products, but did demonstrate the acid intolerance of adult worms and suggests that abomasal microbes may be capable of modulating the secretory activity of the host digestive tract.     

Ostertagia ostertagi infection in the calf: effects of a trickle challenge on the hormonal control of digestive and metabolic function

Metabolic effects of a trickle challenge with the equivalent of 10,000 infective Ostertagia ostertagi larvae per day were investigated in 12 calves allocated to infected, pair-fed control or ad libitum-fed control groups. Changes in hormone levels reflecting abomasal, pituitary and pancreatic function were monitored using radioimmunoassay techniques previously validated for use in cattle. A range of metabolic profile parameters and blood metabolites was also measured. Feed intake of the infected calves began to decline as blood gastrin and pepsinogen levels reached a peak. The depression in appetite recorded in this group was responsible for significant increases in plasma urea and non-esterified fatty acid levels and associated with an increase in growth hormone/insulin ratio. No significant difference in glucagon levels was recorded between groups. A decline in blood albumin values was also shown in the infected group and associated with a drop in nitrogen digestibility. A significant depression in circulating calcium levels was related to either the hypoalbuminaemia or impaired mineral absorption in the intestine. A decrease in plasma cholesterol values in the infected group was associated with changes in digestive function.     

Effects of Ostertagia ostertagi infection on secretion of metabolic hormones in calves.

Effects of Ostertagia ostertagi infection on secretion of insulin, pancreatic glucagon, cortisol, gastrin, and pepsinogen were studied in calves inoculated with 100,000 (group 1) or 10,000 (group 2) O ostertagi infective larvae weekly for 14 weeks. Plasma insulin concentrations in both inoculated groups were lower than those in a non-infected (group 3) control group. The differences between group 1 and group 3 were significant (P < 0.05) at 2 and 12 weeks after initial inoculation. Plasma pancreatic glucagon and cortisol concentrations of groups 1 and 2 did not differ significantly from those of the control group, although plasma pancreatic glucagon concentration was consistently lower in group-1 calves from 4 weeks to end of the study. Plasma pepsinogen and serum gastrin concentrations also increased significantly (P < 0.05) in both groups that received inoculations. We concluded that decreased plasma insulin concentrations are contributory to changes in postabsorptive protein metabolism, and that serum gastrin concentrations are more representative of the pathologic changes in the abomasum than are plasma pepsinogen concentrations.     

Physiological and parasitological responses to nematode infections of fattening cattle in the Western Pampas of Argentina

The epidemiology of nematode infection was studied in fattening grazing cattle from weaning (April 1994) to market at the end of their second autumn (July 1995). Sixty Aberdeen Angus calves of seven months of age were randomly allocated by weight to two groups: GT, treated every three weeks with doramectin (200 mcg/kg); and GI, an infected group, only treated with fenbendazole (7.5 mg/kg) at weaning and on the 1st of October. The two groups were grazed together on contaminated lucerne pastures until July, on 'clean' oat pastures until October and again on contaminated lucerne until the end of the trial. Fecal egg counts (epg), herbage larvae (L3), serum pepsinogen (Pep) and blood eosinophils (Eo) were evaluated monthly. Eight steers were slaughtered for worm recovery, three in July 1994, three in December 1994 and two in July 1995. Grazing feed intake was estimated by fecal output (chromic oxide method)/l-diet digestibility and to measure non-specific response, Brucella antibodies were detected at 11 and 40 days post-vaccination in early winter. Fecal egg counts, Pep and Eo of GI increased (P<0.01) from April to July when there was a moderate-to-high level of infection. Ostertagia, Trichostrongylus and Cooperia were the predominant genera. By late winter, all parameters decreased on oat 'clean' pastures and increased again when cattle returned to moderately infected lucerne. During summer, the parameters measured reflected the negligible numbers of L3 on pastures until early autumn. At this time, increased numbers of L3 were followed by a moderate rise (P<0.01) of epg, Pep and Eo values. During winter, GT showed higher (P<0.04) Brucella antibody IgG titers while feed intake of GI was 24.9% depressed (P<0.02). There were total cumulative weight-gain (WG) differences (P<0.001) between groups (GT=263.1 kg; GI=214.3 kg). During the second autumn, the mean WG of GT steers was 16.6 kg greater (P<0.04) than that of GI. Vaccination titres against Brucella suggested non-specific depression of immunity, while higher Pep and Eo levels in second-year steers may have reflected hypersensitivity reactions.