Studies of the immunomodulatory effects of low-level infection with Ostertagia ostertagi in calves

Possible immunomodulation by low-level infection with Ostertagia ostertagi was studied in 4-month-old calves. Six groups of 4 calves each were subjected to the following regimens: group 1--nonparasitized controls; group 2--nonparasitized, but challenge exposed at day 64 with Brucella abortus strain 19 vaccine (BA) and at day 78 with IV administration of a soluble third-stage larval (L3) antigen preparation of O ostertagi (OAG); group 3--nonparasitized, but challenge exposed at day 78 with 75 x 10(3) L3 of O ostertagi; group 4--continuously parasitized by weekly dosing with 30 x 10(3) L3 of O ostertagi; group 5--continuously parasitized by weekly dosing with 30 x 10(3) L3 of O ostertagi, then challenge exposed on day 64 with BA and on day 78 with IV inoculation of OAG; and group 6--continuously parasitized by weekly dosing with 30 x 10(3) L3 of O ostertagi, then challenge exposed on day 78 with 75 x 10(3) L3 of O ostertagi. Over the initial 10 weeks of the study, nonparasitized calves, (groups 1, 2, and 3) had higher body weight, blood lymphocyte (BL) response to phytohemagglutinin (PHA), and significantly (P less than 0.05) higher feed consumption and lymphocyte numbers, whereas parasitized calves (groups 4, 5, and 6) had higher BL responses to pokeweed mitogen (PWM) and significantly (P less than 0.05) higher neutrophil and eosinophil numbers, plasma pepsinogen (PP) values, and BL response to OAG.(ABSTRACT TRUNCATED AT 250 WORDS)     

Lymphocyte blastogenic responses of calves experimentally infected with Ostertagia ostertagi

Twelve 9-week-old calves were divided into four groups; two groups were maintained helminth-free as controls and the other groups were given Ostertagia ostertagi infective-stage larvae (L3) orally. One group received 100,000 L3 as a single inoculum and the other group received L3 in increasing dosages at weekly intervals for 8 consecutive weeks. The blastogenic responses to concanavalin A (Con A), phytohemagglutinin (PHA), and a soluble larval antigen from O. ostertagi (SLA) of peripheral blood lymphocytes were evaluated using tritiated thymidine incorporation into DNA as a measure of blastogenesis. The responses to Con A of all infected calves were significantly depressed while the responses to PHA were not. SLA, at concentrations of 4 micrograms ml-1 and above, caused blastogenic activity in lymphocytes from uninfected calves. Using SLA at 1 microgram ml-1 in lymphocyte cultures supplemented with autologous serum, an antigen-induced blastogenic response was detected in calves receiving serial inoculations of L3.     

Pathogenesis of simulated natural infections with Ostertagia ostertagi in calves

The possibility of a mucosal hypersensitivity reaction and its relationship to the pathogenesis of simulated natural infections with Ostertagia ostertagi were studied in calves. Four groups of 4 calves each were used. One group was used as noninfected control; a 2nd group was given increasing doses of infective larvae; a 3rd group was given increasing doses of larvae and these were removed by succeeding treatment with an anthelmintic; and a 4th group was given an initial dose of larvae which was then eliminated with an anthelmintic. All calves given larvae became sensitized, as shown by an intradermal skin test. The continuously infected calves had significantly (P less than 0.05) higher fecal egg counts, eosinophil counts, plasma pepsinogen values, and worm burdens and significantly (P less than 0.05) lower lymphocyte counts than did the other groups of calves. These animals also had the most extensive mucosal pathologic changes. The group given intermittent larval challenge exposures followed by an anthelmintic showed decreased lymphocyte values, but these were not significant. Plasma pepsinogen values of this group increased between every challenge exposure and treatment, a 3-day period. This indicated that a mucosal hypersensitivity reaction had occurred in these calves at these times, because they were shown to have been sensitized, and challenge-exposure infections were not present for sufficient time to have produced direct pathologic effects. It therefore seems that a part of the pathologic changes in O ostertagi infections may be the result of the continuous challenge exposure experienced by the animals through a constant intake of larvae from pasture and the intestinal reaction to this challenge exposure.     

Immunosuppression of lymphocyte blastogenesis in cattle infected with Ostertagia ostertagi and/or Trichostrongylus axei

Twenty 4-month-old calves were infected with O ostertagi and/or T axei and the responses to phytolectins were evaluated. Whole blood cultures were incubated with phytohemagglutinin (PHA), concanavalin A (Con A) and pokeweed mitogen (PWM). The blastogenic response was determined by tritiated thymidine uptake with results presented as counts per minute (cpm), stimulation indices (SI) and a mononuclear cell responsive index determined by dividing the phytomitogen induced cpm by the absolute mononuclear cell number per ul. The control group results were adjusted to 100 percent and changes in the percentage difference by the parasitized calves was determined. There was a decline in lymphocyte responsiveness to PHA beginning at the time of infection. Significant depression of responses to PHA was observed in all parasitized calves 8 weeks after infection although clinical signs of parasitism did not occur. Lymphocyte responses to PW, were not different in infected calves from the control, although the O ostertagi group had significantly higher PWM mean upon than the calves infected with T. axei. A slight depression in response to Con A was also observed at 8 weeks after infection followed by a significant increase after 10 weeks. The immunosuppression appeared to be a feature of gastrointestinal parasitism and related to infections with O. ostertagi and/or T. axei.     

Efficacy of ivermectin delivered from a sustained-release bolus against inhibited early fourth-stage larvae of Ostertagia ostertagi and other nematodes in cattle.

The anthelmintic efficacy of ivermectin (IVM) delivered from a sustained-release (SR) bolus was evaluated against natural infections with gastrointestinal tract nematodes in 12 crossbred beef heifers in spring. The 12 calves were randomly allotted to 2 groups of 6 calves each. Group-1 calves were treated with an SR bolus designed to deliver 8 mg of ivermectin/d. Group-2 calves were nontreated controls. Cattle groups were kept in separate concrete-floored pens (grass hay nutrition) and slaughter was performed at 35 days after treatment. Fecal egg counts for group-1 calves remained zero after treatment, except for detection of less than 1 egg/g of feces in 1 calf at the time of slaughter; counts in nontreated calves increased. Mean and range of Ostertagia ostertagi inhibited larvae in nontreated calves were 27,093 and 10,622 to 56,368, respectively. Efficacy of the IVM SR bolus was 100% against O ostertagi developing fourth-stage larvae (L4) and inhibited early L4, Haemonchus placei adults, Cooperia punctata and C spatulata adult males, Cooperia spp adult females, Cooperia spp L4, Trichostrongylus colubriformis adults, Bunostomum phlebotomum adults, and Oesophagostomum radiatum adults. Efficacy for O ostertagi and T axei adults was 99.9%. Numbers of nontreated calves infected with C pectinata adult males and Oes radiatum L4 were too low to evaluate efficacy. Calves treated with the IVM bolus gained 10.2 kg, whereas nontreated calves lost 1.8 kg. Abomasal lesions were clearly greater in nontreated calves on the basis of index comparisons of abomasal weight and total live weight and gross pathologic features.     

Direct anthelmintic and immunostimulatory effects of oral dosing semi-purified phytohaemagglutinin lectin in sheep infected with Teladorsagia circumcincta and Trichostrongylus colubriformis

Lectins are plant secondary compounds that can have anthelmintic properties in vitro. In particular, the phytohaemagglutinin lectin extracted from Phaseolus vulgaris has been shown to inhibit the feeding of Trichostrongylus colubriformis and Teladorsagia circumcincta L1 larvae. However, little is known about the potential anthelmintic properties of this lectin in vivo and its suitability to control gastrointestinal parasite infections in lambs. In a 2 × 2 study, lambs were either orally dosed, or not, with 2.3mg semi-purified PHA lectin per kg live weight (LW) per day, whilst concurrently infected, or not, with 1000 T. circumcincta and 1000 T. colubriformis L3 infective larvae per day for 42 days. There were no adverse clinical effects observed with this dose of PHA lectin. Although worm burdens were similar, animals dosed with PHA lectin had reduced concentration of nematode eggs in the faeces compared with their non-lectin dosed counterparts (P=0.026), suggesting that there may be a direct effect of PHA lectin on parasite fecundity. Irrespective of infection, PHA lectin had immune-stimulatory properties with increased eosinophillia in both abomasal and small intestine tissue sections taken at slaughter on day 42 (P<0.02 for both) and a tendency for decreased ability of Teladorsagia larvae to penetrate abomasal tissue explants (P=0.06). Compared with infection alone, concurrent PHA lectin dosing and infection further increased the number of eosinophils (P<0.01), PAS-positive (mucin-producing cells) (P=0.03) and tended to increase the number of T helper cells (P=0.06). No interactions were observed for cell populations in small intestine tissue sections. These results suggest PHA lectin could have two possible modes of action against T. circumcincta and T. colubriformis, a direct anthelmintic effect on nematode fecundity and an indirect effect through enhancing local immune responses in the host.     

Larval migration inhibition activity in abomasal mucus and serum from calves infected with Ostertagia ostertagi

The present study investigated whether abomasal mucus from calves naturally infected with gastrointestinal nematodes possessed larval migration inhibition (LMI) activity in vitro, and whether LMI activity was greater in mucus from previously immunised animals, compared to primary infected and uninfected calves. LMI activity was also assessed in serum from calves during both natural and artificial Ostertagia ostertagi infections, in an attempt to monitor the development of acquired immunity. Both abomasal mucus and serum exhibited larval paralysing activity. Although the LMI capacity of the abomasal mucus was very variable, the highest paralysing activity was consistently observed in mucus from previously immunised calves. LMI activity in serum increased significantly during both artificial and natural Ostertagia infections. After a challenge infection, sera from immunised animals showed a significantly higher LMI capacity, compared to previously uninfected calves. Moreover, serum LMI activity was significantly negatively correlated with Ostertagia worm counts after the challenge infection. The present results suggest that LMI activity in serum and/or abomasal mucus reflects a protective immune response against O. ostertagi in the abomasal mucosa.     

Spontaneous and mitogen-induced RNA synthesis by blood lymphocytes from bovine leukemia virus-infected and normal cows

Peripheral blood lymphocytes (PBL) from normal and bovine leukemia virus (BLV)-infected cattle were prepared by density gradient technique and incubated with and without phytohaemagglutinin (PHA) and pokeweed mitogen (PWM). RNA synthesis was determined at different periods of incubation by 3H-uridine incorporation. PBL from BLV-infected cows with persistent lymphocytosis (PL) showed the highest spontaneous RNA synthesis. PBL from BLV-infected cows with normal lymphocyte counts synthesized more RNA than cells from normal animals. Decreased mitogen responses were observed in PBL from infected cows with PL in comparison to normal and BLV-infected cattle without PL. PHA and PWM did not show significant differences in their degree of stimulation of RNA synthesis.     

Influence of isoprinosine on bovine herpesvirus type-1 infection in cattle

A study was conducted to determine the in vivo efficacy of isoprinosine (ISO) in calves infected with bovine herpesvirus type-1 (BHV-1). Calves were infected with BHV-1 on day 0 and received ISO daily for 14 days. Clinical signs of disease, shedding of BHV-1, lymphocyte proliferative responses to mitogens, interleukin-2 production, and alveolar macrophage bactericidal activity were monitored during the study. Rectal temperatures were increased (P less than 0.05) in BHV-1 and ISO-BHV-1 calves at days 3 to 7 postinfection (PI). Isoprinosine did not influence BHV-1 shedding in calves. Lymphocyte proliferative responses to phytohemagglutinin (PHA) were lower (P less than 0.01) in BHV-1 calves when compared to control or ISO calves at day 4 PI, but ISO did not ameliorate this effect. Interleukin-2 activity was greater (P less than 0.05) in ISO-BHV-1 calves on days 4 and 8 PI in PHA-stimulated lymphocytes and on day 8 PI in concanavalin A-stimulated lymphocytes when compared to control, ISO or BHV-1 calves. Isoprinosine treatment of BHV-1-infected calves tended to decrease alveolar macrophage bactericidal activity. These data suggest that ISO does not reverse BHV-1 suppression of lymphocyte proliferation, but may enhance IL-2 production in BHV-1 infected calves.     

Persistence of dead Ostertagia ostertagi in the abomasal mucosa following anthelmintic treatment

Anthelmintic trails, conducted with albendazole, fenbendazole and ivermectin for efficacy against gastrointestinal nematodes, principally inhibited early fourth larval stages of Ostertagia ostertagi in naturally infected cattle. Cattle wee slaughtered seven to 20 days after treatment. O ostertagi was the predominant abomasal nematode recovered with occasional small numbers of Haemonchus species and Trichostrongylus axei. Control calves uniformly had very large O ostertagi infections, primarily early fourth stage larvae. Viable surviving worms and variable numbers of dead and degenerate worms were recovered in abomasal contents and washings. These O ostertagi larvae and adults were characterised by adherent debris or proteinaceous material, degenerated cuticles and distortion of internal structures. This study demonstrated the necessity for proper timing of slaughter for anthelmintic trial evaluation to allow clearance of dead nematodes, specifically O ostertagi larvae which are sequestered in the abomasal glandular tissue. Nematode collection within seven to 12 days after treatment will include dead and degenerate larval nematodes. The peripheral coating of larvae was suggestive of the Splendore-Hoeppli effect which has been associated with immunological responsiveness. The antigenic stimulus for this material and the lymphocyte and eosinophil infiltration was suspected to be early fourth stage O ostertagi larvae within the mucosa but was not identified definitively.     

Epidemiology, effects and control of nematode infections on Zebu crossbred, Hereford and Hereford X Brahman calves of Argentina's western pampas

Gastrointestinal parasitism of Zebu crossbred (Z), Hereford (H), and Hereford X Brahman (HB) weaned steer calves was observed from March 1985 to May 1986. Three groups of 60 calves were randomised and in each group the three breeds were equally present. Calves in GI were untreated, in GII were treated strategically and in GIII treated monthly with anthelmintic. The effects of anthelmintic treatments on the growth of cattle were assessed by comparing mean live-weight gain (MLG) responses of treated and untreated calves grazing the same permanent pasture. Worm recovery and egg counts increased from autumn to winter. After abundant July rainfalls, herbage infective larvae (L3) and worm burdens (mainly Ostertagia) reached the highest levels and Z calves presented symptoms of clinical parasitism. After high availability of L3 in spring, L3 and egg counts decreased to the end of the study. From the end of winter until summer a high percent of inhibited Ostertagia EL4 was seen. Blood samples showed reduced serum copper lower levels for GI. MLG responses of Z treated calves were significantly higher during the autumn-spring and summer-autumn periods. MLG responses of treated H calves were significant only during the winter-spring period. For HB calves there were no significant MLG responses during any period. The highest monthly treatment response was observed during winter and early spring. All strategic treatments gave significant MLG responses in spring and late summer. Initial effects of parasitism, from autumn until early spring had the greatest consequences in current and later productivity. For Z calves herbage L3, worm burdens, egg counts and MLG responses were higher than for H and HB calves. The Z breed were significantly heavier (+22.7%) at the end of the experiment in GIII than in GI. Similar trends were recorded, significantly, for H breed (+17.7%) and not statistically significant for HB (+12.7%). Under our temperate conditions it seems that H and HB calves showed greater capacity to resist nematode infection than Z calves.     

Experimental and natural infection of calves with Bunostomum phlebotomum

An experiment was conducted to determine the efficiency of a method of experimental infection of weaner beef calves with Bunostomum phlebotomum and to compare such infection with that established by natural infection. Six calves, maintained on a concrete-floored pen, were inoculated with B. phlebotomum L3 by placing the larval inocuulum, in small volume, in the outer chamber of the ear while the animal was restrained for 18 min. Inoculation doses of 20, 30, 40, 50, 60, and 80 thousand L3 were used. Six other calves were grazed on pasture known to be heavily contaminated with hookworm. All animals were killed 72 days after experimental infection and 93 days after initial exposure to pasture infection. The experimental and naturally-infected calves became patent at 55 and 66 days, respectively, after exposure to L3. Red blood cell counts and hemoglobin values were markedly depressed in both groups and lowest values coincided with onset of patency. There was difference in liveweight changes, but both groups lost weight during the prepatent period of infection and gained weight with the onset of patency. The largest number of hookworms was established at the 30000 L3 inoculation level; little or no establishment was observed at the 2 highest levels. Sizeable adult hookworm burdens were established in 4 out of 6 pastured calves. Intestinal pathology was generally more severe in experimentally-infected calves, consisting of a thickened mucosa and masses of punctate, hemorrhagic foci. Pastured calves also acquired large burdens of Ostertagia ostertagi, particularly inhibited early fourth-stage larvae. Moderate to severe abomasal pathology and elevated plasma pepsinogen were associated with ostertagiasis in the pastured calves. The experimental infection method is efficient in establishing high levels of B. phlebotomum infection in calves currently or previously infected with other gastrointestinal nematodes.     

mRNA profile of Nellore calves after primary infection with Haemonchus placei

Haemonchus parasites are responsible for many losses in animal production. However, few studies are available, especially of zebu cattle. In this study, we investigated mRNA differences of immune response genes in na?ve Nellore calves infected with Haemonchus placei, relating these differences to patterns of cellular infiltrate. Calves were infected with 15,000 H. placei L3 larvae and after 7 days lymph node and abomasum tissues were collected. IL-2, IL-4, IL-8, IL-12, IL-13, IFN-γ, MCP-1, lysozyme, pepsinogen and TNF-α genes were evaluated by qPCR. Mast cells, eosinophils and globular leukocytes were counted by abomasum histology. In the infected group, IL-4, IL-13 and TNF-α were up-regulated in the abomasal lymph node. In the abomasum, IL-13 increased and TNF-α was down-regulated (p<0.05). No differences were detected for mast cells and eosinophil counts in abomasal tissue (p>0.05). We conclude that for this infection time, there was Th2 polarization but that cellular infiltrate in abomasal tissue takes longer to develop.     

Cell-mediated immune response in calves to single-dose, trickle, and challenge infections with Fasciola hepatica

A peripheral blood mononuclear cell (PBMC) proliferation assay was used to study the cell-mediated immune response in eight calves experimentally infected with Fasciola hepatica. Hypersensitivity-related eosinophil and mast-cell responses were also assessed. The primary infection of 500 metacercariae was administered either as a single-dose or as a trickle infection over a 4-week period. Calves were challenge-infected 4 months later with 100 metacercariae and slaughtered 24 weeks postprimary infection. Skin eosinophil counts (SEC) were determined prior to infection on the basis of the intradermal reaction (IDR) to phytohaemagglutinin (PHA). These counts correlated negatively with the mean fluke length but not with the fluke burden found at necropsy. At the end of the experiment, non-specific (PHA) and specific (excretory-secretory parasite, products, FhESAg, and whole-worm extract, FhSomAg) immediate type hypersensitivity IDR were elicited in contrast to delayed type hypersensitivity (DTH) responses. The SEC correlated with blood eosinophilia but not with parasite parameters. These findings suggest that the eosinophil response does not correlate clearly with the development of resistance to F. hepatica infection in cattle. A specific mononuclear cell response to FhSomAg was detectable as early as 7 days after infection in both infected groups, being significantly higher during the very early migratory phase of the juveniles in the single-dose infected calves than in the trickle infected calves. This response remained significantly higher in infected groups than in the control group throughout the experiment. Challenge elicited a significant proliferative response, less pronounced than after primary infection. No production of gamma-interferon (INF-gamma) was recorded 3 weeks after challenge. At necropsy, the mean number of flukes recovered was similar in both infected groups, suggesting that the rate at which the infection is administrated has no effect on protective immunity. Hepatic lesions, similar in both infected groups, were characterised by marked eosinophil and mast-cell infiltration. Liver biopsies were performed and their diagnostic value is discussed. All results suggest that F. hepatica infection predominantly induces a Type-2 response in cattle, and that this response has little protective effect.     

Lymphocyte alterations in zinc-deficient calves with lethal trait A46

Lymphocyte numbers and activities were evaluated at 2, 4, 8 and 12 weeks of age in two calves with lethal trait A46 (A46), a genetic disorder affecting intestinal zinc absorption. Plasma zinc concentrations declined to subnormal by 3 weeks of age, after which anorexia, diarrhea, alopecia and hyperkeratosis occurred. Lymphocyte response to phytohemagglutinin-P (PHA), concanavalin A (Con A) and pokeweed mitogen (PWM) stimulation was variably reduced. CD4+ T-lymphocytes were subnormal on at least one observation period following onset of zinc deficiency, and relative numbers of B lymphocytes were decreased at 8 weeks. Secondary antibody responses to bacteriophage phi X 174 were significantly reduced. The results demonstrate that calves homozygous for the A46 trait have normal numbers of functional lymphocyte subpopulations at birth, and that the activity of their lymphocytes is altered once the calves become zinc deficient.     

Experimental Haemonchus contortus infection in three breeds of sheep

Nine- to 10-month-old Florida Native, St. Croix and Dorset/Rambouillet sheep were infected with Haemonchus contortus. The primary infection was terminated by anthelmintic treatment 9 weeks later and animals were then reinfected with H. contortus. Dorset/Rambouillet sheep showed higher fecal egg counts and decreased packed cell volumes and plasma protein levels compared with the other two breeds during the primary infection. However, no breed differences were found in total worm burdens in either primary or secondary infection. Differences between breeds were also not detected in lymphocyte responsiveness to parasite antigen, H. contortus specific mucosal antibody levels, numbers of abomasal mast cells and globule leukocytes or abomasal histamine levels.     

Efficacy of moxidectin against gastrointestinal nematodes of cattle.

Three groups of 11 naturally infected crossbred beef calves were injected subcutaneously with moxidectin 1 per cent injectable at 0.2 or 0.3 mg moxidectin/kg bodyweight or with the unmedicated vehicle. Nematode infections had been acquired during grazing from December to April. Based on the faecal egg counts and total worm counts of the control calves at necropsy (11 to 13 days after treatment) most of the calves had heavy parasitic burdens. Ostertagia ostertagi was predominant and the mean numbers of adults, developing fourth stage larvae (L4) and inhibited early L4 were 45,906, 10,061 and 68,918, respectively. Haemonchus placei and Trichostrongylus axei were also present in the abomasa. Three species of Cooperia, Oesophagostomum radiatum L4 and T colubriformis adults were found in the intestinal tract. Both dosages of moxidectin were equally effective (P < 0.05) against all the abomasal nematodes (99.9 to 100 per cent) and the intestinal tract nematodes (99.4 to 100 per cent). No adverse reactions to the moxidectin treatment were observed. Abomasal pathology characteristic of heavy O ostertagi infection was observed in the control calves, but not in the treated calves.     

Synergistic influence of Ostertagia ostertagi and Trichostrongylus axei on Ostertagia ostertagi larval inhibition and abomasal lesions in cattle

Parasite-free 4-month-old calves were inoculated with Ostertagia ostertagi and/or Trichostrongylus axei followed 6 weeks later by increasing doses of O ostertagi for 8 weeks. Clinical signs of parasitism, fecal egg counts, and plasma pepsinogen concentrations were monitored, and gross lesions and parasite burdens were determined postmortem. Clinical signs of parasitism were not observed and weight gains were not affected in experimentally infected calves. In calves infected with O ostertagi, mean plasma pepsinogen concentrations were greater than for control calves and were diagnostically significant 4 weeks after inoculation and during the last 4 weeks of serial inoculations with O ostertagi. In calves that were given O ostertagi and T axei, abomasal pH was significantly increased, and abomasal lesions were more pronounced than in control calves or in calves inoculated with only O ostertagi or T axei. Abomasal lymph nodes were enlarged in all parasitized calves; other lymph nodes in the calves inoculated with both O ostertagi and T axei were usually smaller than in calves inoculated with only O ostertagi or T axei. Numbers of O ostertagi-inhibited larvae were small in all inoculated calves, but the percentage inhibition was significantly greater in calves inoculated with both O ostertagi and T axei. The percentage inhibition was 3.53% for the O ostertagi-inoculated calves and 7.07% for calves inoculated with both O ostertagi and T axei. These percentages indicated a synergistic effect of concurrent abomasal parasitism, whereas a synergistic effect on T axei worm burden was not observed. The low percentage of larval inhibition indicated that factors other than host resistance are involved in naturally occurring pretype II ostertagiosis.     

Comparison of abomasal emptying in neonatal calves with a nuclear scintigraphic procedure.

The purpose of the present study was to demonstrate that nuclear medicine technology allows observation of the effect that milk clotting has on abomasal emptying in the living neonatal calf. Scintigraphic evaluation of abomasal emptying was carried out in 6 healthy male Holstein calves. The calves were fed 10% of their body weight daily as whole cow's milk that was divided equally and consumed as 2 feedings via a nipple bottle. One day before the nuclear scintigraphic procedure, the calves were randomly fed whole cow's milk, or an oral rehydration solution (ORS) containing bicarbonate and high levels of soluble fibre was fed for 3 consecutive feedings an hour before the portion of milk. For each calf, both feeding programs were repeated twice at a one-week interval. Immediately following administration of the 99mTC-sulfur-colloid-containing milk, the calves were imaged with the gamma camera positioned lateral and ventral to the abomasum. Additional right lateral and ventral views of the abomasum were collected at 15, 30, 45, 60, 90, 120, 150, 180, 210, and 240 min after administration of the radionuclide. Blood glucose determination were performed at one-hour intervals for 7 h after feeding milk to evaluate milk digestibility in both feeding programs. No significant differences in the results of the glucose absorption test or in the radionuclide counts of the abomasum were found between both feeding programs. Scintigraphic evaluation of abomasal emptying was found to be a useful technique for visualization of milk clotting and to test the effect of an ORS on milk digestibility.     

Cellular profiles in the abomasal mucosa and lymph node during primary infection with Haemonchus contortus in sheep

Cellular changes in the abomasal tissue and draining abomasal lymph nodes were examined after primary infection of lambs with Haemonchus contortus for 3, 5 or 27-36 days.Infection with H. contortus larvae resulted in a rapid and selective increase in the percentage of CD4(+) T-cells in the abomasal lymph node at 3 days post-infection (PI). By 5 days PI, the lymph node weight had increased two-fold; however, the percentage of lymphocyte populations in the abomasal lymph node resembled that seen in uninfected sheep. Lymph node weights remained at increased levels in the adult nematode infected sheep and down-regulation of B-cell surface markers (sIg and MHC Class II) was apparent in this group. Significant increases in the percentage of CD4(+) T-cells co-expressing MHC Class II, but not CD25, were observed in the larval infected groups except in adult nematode infected sheep. Increased numbers of eosinophils, CD4(+), gamma delta(+) T-cells and B-cells were found in the abomasal tissue by 5 days PI, but no further increases in these cell populations were observed in the adult nematode infected group. In contrast, the level of both lamina propria and intraepithelial mast cells observed in the abomasal mucosa was highest in the sheep carrying an adult nematode burden. These findings indicate that sheep are able to generate an early immune response to infection with H. contortus larvae, characterised by the activation of CD4 T-cells and B-cells in the draining lymph nodes and recruitment of eosinophils, CD4(+) and gamma delta-TCR,WC1(+) T-cells and B-cells in larval infected tissues. However, these changes do not seem to be maintained during infection with the adult parasite where increases in mast cell numbers dominate the local response, indicating that different parasite stages may induce distinct and possibly counteractive immune responses.