The effects on merino lambs of chronic infection with Trichostrongylus rugatus

Four groups of six 5-month-old merino lambs were exposed to infection with 12,000 larvae of Trichostrongylus rugatus per week and one group was killed at each of 3, 6, 9 and 12 weeks. Numbers of adult worms increased in an approximately linear manner throughout the experiment; the percentage of larvae developing to adults remained between 35 and 25% throughout the course of the experiment. Sub-total villus atrophy was evident in the anterior small intestine of each group of infected sheep and significant reductions occurred in the plasma concentrations of albumin and phosphate. Serum globulin and calcium concentrations rose in infected animals. No clinical signs were observed in infected sheep, no alteration in bone histology was detected and no decrease in growth rate occurred.     

Effects of temperature and relative humidity on development and survival of the free-living stages of Trichostrongylus colubriformis, T. rugatus and T. vitrinus

Temperatures of 4 and 10 degrees C reduced the rate of hatching of eggs and the development of the pre-infective stages of T. colubriformis and T. rugatus, but not T. vitrinus. Eggs of T. rugatus hatched at 4 degrees C, but did not develop to the infective stage. At 20 and 30 degrees C, no differences were detected between species in egg hatching or the development of infective larvae. Third-stage larvae of T. rugatus survived for significantly longer periods of time at 33 and 56% relative humidities at a temperature of 20 degrees C than did either T. colubriformis or T. vitrinus. At 30 degrees C, differences were less marked, but T. rugatus and T. colubriformis survived longer than T. vitrinus. The results are discussed in relationship to the known distribution of these species in eastern Australia.     

Regulation of Trichostrongylus vitrinus and T colubriformis populations in naturally infected sheep in the Netherlands

During a study on the epidemiology of gastrointestinal helminth infections in sheep at the University of Utrecht (1974 to 1977) information was obtained on the population regulation of small intestinal Trichostrongylus species. Comparison of the cumulative burdens in tracer lambs with those in flock lambs showed an accumulation of Trichostrongylus species in the latter until autumn followed by an expulsion of worms. The dominating species, T vitrinus, was expelled earlier than T colubriformis. The proportion of inhibited early third stage Trichostrongylus species larvae in tracer lambs was low, but it tended to increase in autumn. In most flock lambs the levels of inhibition were also low. In non-lactating ewes, populations consisted predominantly of inhibited early third stage larvae but when adults were present they tended to be mainly T colubriformis. Similar populations were seen in some lactating ewes. In other lactating ewes Trichostrongylus species populations with low proportions of inhibited early third stage larvae which were dominated by T vitrinus, indicated a relaxation of resistance around parturition.     

Selection of ivermectin-resistant Trichostrongylus colubriformis in lambs

A mixed population of Haemonchus contortus, Teladorsagia circumcincta and Trichostrongylus colubriformis was passed for four generations through lambs. In successive passages and on Day 6 (D6), lambs were dosed subcutaneously with ivermectin at 100, 200, 250 and 225 micrograms kg-1 body weight, respectively. No larvae of H. contortus were recovered after the first passage and following the third drug exposure only T. colubriformis was present. Larvae of the selected strain of T. colubriformis developed to third stage, sheathed larvae in vitro in concentrations of ivermectin that inhibited development of larvae from the original strain of T. colubriformis. A definitive control test was then run with 45 lambs, to compare this selected strain to the original strain. The treatments were 150 micrograms kg-1 subcutaneous (SC) D6, 200 micrograms kg-1 SC D6, 200 micrograms kg-1 intraruminal (IR) D6 and 200 micrograms kg-1 SC D21; all treatments were compared to non-treated controls, selected or original strain as applicable. The D6 IR and D6 SC 150 micrograms kg-1 treatments were greater than 99% effective in both strains, although the D6 SC 200 micrograms kg-1 treatment was 85 and 48% effective in the original and selected strain, respectively. The D21 200 micrograms kg-1 was only 58% effective against the selected strain. There was significant (P less than 0.001) differences between the original and selected strains of T. colubriformis for both fecal egg output and worm counts. The exposure of four generations of immature T. colubriformis to ivermectin selected for partial resistance.     

Effect of concurrent infection with Ostertagia circumcincta and Trichostrongylus vitrinus on the performance of lambs

The interaction between Ostertagia circumcincta and Trichostrongylus vitrinus was investigated in lambs continuously infected over 12 weeks. Four groups of seven lambs were given either zero, 1000 T vitrinus larvae per day, 2500 O circumcincta larvae per day or infected concurrently. Overall liveweight gain was reduced by 17, 20 and 30 per cent in T vitrinus, O circumcincta and concurrent infection, respectively. T vitrinus infection significantly lowered serum phosphorus concentrations by week 4. In the concurrent infection the decline in phosphorus concentration was more gradual and only differed significantly from the controls during the final four weeks of the trial. There were no significant differences between the total numbers of T vitrinus or O circumcincta recovered from single or combined infections. The lack of an additive effect on performance may be due to a delayed establishment of T vitrinus in the presence of O circumcincta.     

Effects of dietary molybdenum on nematode and host during Trichostrongylus vitrinus infection in lambs.

The addition of molybdenum (0.05 mmol kg-1 dry matter) to the diet of lambs exposed for four weeks to a trickle (2500 third stage larvae per day) infection with Trichostrongylus vitrinus reduced the number and length of adult worms retrieved from the small intestine 11 days later: both effects were particularly marked in female worms from female lambs (P less than 0.01). Worms from lambs given molybdenum contained less proteinase enzyme activity and secreted less proteinases in culture irrespective of the sex of the host. Pathogenicity was not attenuated by molybdenum. Damage to the intestinal mucosa was severe in both dietary groups but infected females given molybdenum developed lower plasma albumin concentrations and lighter dressed carcases than those not given molybdenum. Neither the effects on the parasite nor those on the host could be attributed simply to molybdenum-induced copper depletion, using conventional measures of copper status. Molybdenum may be toxic to T vitrinus but may also facilitate or enhance the inflammatory process limiting larval establishment or increasing parasite rejection.     

Effect of prolonged subclinical infection with Trichostrongylus colubriformis on the performance and nitrogen metabolism of growing lambs

Ten 5-month-old lambs, reared free of parasites and prepared with abomasal and ileal cannulas, were paired and given rations of pelleted dried grass by means of continuous feeders. From 7 months of age one of each pair was dosed daily with 2500 Trichostrongylus colubriformis larvae for 34 weeks. An untreated animal served as a pair-fed control to each infected lamb. During 5 experimental periods (pre-infection, and Weeks 6-8, 12-14, 18-20 and 29-34 of dosing) measurements were made of the amounts of N leaving the small intestine, the digestibility of 35[S]-labelled microbial protein in the small intestine, and the amounts of plasma protein leaking into the digestive tract. The infection caused a growth check (P less than 0.05) from Week 6 to approximately Week 13 of dosing. This resulted from inappetence, but also from a disruption in the N digestion and metabolism of the lambs. There was a marked eosinophilia at this time. Measurements of ileal flow rates during Weeks 6 and 12 indicated that significantly more N flowed to the terminal ileum of infected lambs during this period (Week 6, 3.1 g N day-1 P less than 0.05; Week 12, 5.0 g N day-1 P less than 0.001). Plasma N leakage into the intestine during Weeks 8 and 14 was higher in infected lambs (1.2 g N day-1 P less than 0.01 and 0.9 g N day-1 P less than 0.05, respectively) but the digestibility of 35[S]-labelled microbial protein was depressed below that of the controls during the later part of this period, i.e. during Week 13. Continued larval challenge beyond this point caused a less severe nutritional penalty. Voluntary feed intake increased and the animals started to gain weight. The amount of extra N flowing to the terminal ileum of infected lambs was less at Week 18 of dosing (3.5 g N day-1 P less than 0.01) than it had been at Week 12. Similarly, extra plasma N leakage into the tract of infected lambs was less during Week 20 (0.26 g N day-1) than during Week 14. There was however, still a detectable reduction in the coefficient of digestibility of 35S microbial protein in infected lambs during Week 19. By Weeks 29-34 the only detectable difference between the infected and control lambs was in their eosinophil counts.     

Development of immunity to incoming radiolabelled larvae in lambs continuously infected with Trichostrongylus vitrinus

Infective third stage larvae (L3) of Trichostrongylus vitrinus were radiolabelled with 75 selenium by a method which did not affect their viability. Three groups of six-month-old lambs were infected daily with 1000 L3 for four, eight and 12 weeks, respectively. After each period, one of those groups (n = 5) and a group (n = 4) of worm-free controls were challenged with three consecutive daily doses of 1000 radiolabelled L3, killed 10 days after the first dose, and their worm burdens examined. After four weeks of continuous infection partial immunity to the establishment of challenge L3 was apparent, and by eight and 12 weeks, with the exception of one sheep, there was almost total resistance to incoming worms. Immunity was also expressed as an inhibition of the development of established worms.     

Influence of two levels of concurrent infection with Ostertagia circumcincta and Trichostrongylus vitrinus on the growth performance of lambs

Groups of seven lambs were infected on five days each week for 11 weeks with either 1000 or 2000 Trichostrongylus vitrinus larvae alone or concurrently with 1500 or 2500 Ostertagia circumcincta larvae and their growth performance compared to that of worm-free controls. The cumulative liveweight gain of the infected groups was significantly lower than that of the controls but the gain of the concurrently infected lambs did not differ significantly from the respective T vitrinus group. There were no significant differences in the numbers of T vitrinus or O circumcincta recovered from the single and concurrently infected groups, although there was a tendency for lower mean T vitrinus burdens at the higher level of ostertagia/trichostrongylus infection. The distribution of T vitrinus along the small intestine was similar in single and concurrently infected lambs. The reason for the lack of a marked additive effect on growth rate with concurrent infection is discussed in relation to changes in the concentration of two plasma constituents and lesions in the gastrointestinal tract.     

Chronic infection with Trichostrongylus vitrinus in sheep. Pathological changes in the small intestine

Eight four-month-old sheep were infected daily for 90 days with 2500 Trichostrongylus vitrinus larvae. Infections were patent by week 3; mean faecal egg counts reached a peak of 645 epg at week 5, then subsequently declined. Mean worm burden at slaughter was 6100 (range 60--25,080). Two sheep had visible lesions in the small intestine at slaughter. In both sheep, numerous shallow red depressed areas extended for 5 to 7 metres distal to the pylorus while, in one, more extensive irregular depressed areas occupied the first 1.5 metres. Microtopographical examination and scanning electron microscopy showed that these affected areas were devoid of villi and contained numerous worms embedded in the mucosa. Histological examination revealed many similarities with the lesions caused by T colubriformis. Subtotal villous atrophy and crypt elongation were present in the intestines from five of the remaining six sheep, and in these the mucosa contained numerous mononuclear cells and intraepithelial globule leucocytes.     

Chronic infection with Trichostrongylus vitrinus in sheep. Some effects on food utilisation, skeletal growth and certain serum constituents

The effect on food utilisation and skeletal growth of daily infection for 90 days with 2500 Trichostrongylus vitrinus larvae was investigated. Controls were killed initially or fed ad libitum. Food intake, digestibility of dry matter and nitrogen and content of body fat, protein, water and minerals were determined, and selected bones examined chemically and histologically. Serum constituents were determined weekly. Digestibility of food dry matter and nitrogen were reduced during weeks 3 and 5 after initial infection but subsequently recovered. Deposition of fat, protein, calcium and phosphorus in the body was reduced by 36, 58, 67 and 78 per cent respectively. Gross efficiency of utilisation of metabolisable energy was reduced by 36 per cent. Bone changes were associated with a reduction in osteoblastic activity. Hypoalbuminaemia, hyperglobulinaemia and hypophosphataemia were present in infected sheep. There were no significant changes in serum calcium or urea concentrations. It is concluded that the effects of continuous intakes of T vitrinus and T colubriformis larvae on host metabolism are essentially the same.     

Trichostrongylus colubriformis and Haemonchus contortus infections in light bodyweight Merino lambs

OBJECTIVE; To show that low bodyweight is a predisposing cause of high Trichostrongylus colubriformis and Haemonchus contortus burdens and egg counts in Merino lambs. DESIGN: A comparison was made, among lambs of different bodyweights, on the effect on immunity of a primary or secondary viable infection with T colubriformis or H contortus larvae. PROCEDURE: Sixty-one Merino lambs, 1 or 6 months of age, were penned indoors and given primary, or both primary and secondary, infection of T colubriformis or H contortus. Faecal egg counts, worm counts and parasite-specific immunoglobulin concentrations were examined for their relationships with bodyweight. RESULTS: Bodyweight at the start of a primary infection was correlated with worm burden, worm fecundity and jejunal IgA antibody concentration. Merino lambs weighing less than 23 kg at the time of first exposure to T colubriformis or H contortus had impaired ability to develop protective mucosal immunity and to resist homologous challenge. CONCLUSION: If the goal is to ensure that lambs develop immunity before weaning, then every endeavour should be made to achieve the combination of critical bodyweight and exposure to moderate levels of nematode infection as soon as possible.     

Susceptibility of the pre-parturient ewe to infection with Trichostrongylus vitrinus and Ostertagia circumcincta

During two successive years, groups of pregnant ewes and non-pregnant worm-free sheep were given anthelmintic, challenged one week later with 10,000 infective larvae of either Trichostrongylus vitrinus or Ostertagia circumcincta and killed two, four and six weeks after challenge. All of the ewes were killed within three days of their predicted lambing date. There was no annual variation in the response of the ewes to infection with either species. Compared to adult naive animals, ewes expressed resistance to infection with T vitrinus at all times, with significantly lower worm burdens composed largely of inhibited third stage larvae. Ewes challenged with O circumcincta were fully as susceptible as naive animals with regard to the size and stage of development of their worm populations. Mechanisms regulating the numbers of adult O circumcincta were operative in the adult naive animals but not in the ewes.     

Dietary impacts on the resistance of Merino lambs to Trichostrongylus colubriformis

AIM: To evaluate and compare the effects of a variety of diets on the resistance of young, lightweight Merino lambs to repeated low-dose infections of Trichostrongylus colubriformis.

METHODS: Ninety-six 12-week-old lambs were fed balanced or unbalanced diets, and given primary (at 19 weeks of age) or both primary (at 12 weeks) and secondary (at 19 weeks) infections of T. colubriformis. Suboptimal diets were low in total intake, rumen undegradable protein, total protein, readily available carbohydrate or minerals, or supplemented with cod liver oil. Liveweight was monitored fortnightly, protection was assessed from worm counts at the end of the study (25 weeks) and faecal nematode egg counts (FEC) conducted weekly, and mast-cell response assessed at the conclusion of the study from concentration of mast-cell protease in the jejunum. Concentrations of glucose in plasma were measured at 21 weeks.

RESULTS: Diets unbalanced for total protein, readily available carbohydrate, Mo or cod liver oil each increased the worm counts after 6 weeks of secondary infection (at 25 weeks of age) and FEC during secondary infection. Low intake of a highnutrient diet restricted liveweight gain but did not affect the development of resistance, as indicated by worm counts and FEC. Defi ciency of available carbohydrate resulted in reduced liveweight gain and reduced concentration of glucose in plasma. There was a reduction in jejunal mast-cell protease concentration in sheep fed unbalanced rations.

CONCLUSIONS: The parasitology results and reduction in the concentration of jejunal mast-cell protease observed in sheep fed unbalanced rations suggest that the nutrient defi ciencies and the fi sh-oil supplement may have acted via inhibition or retardation of the host's acquired immune response. Diets unbalanced for a range of components adversely affected the acquired protective response to T. colubriformis to a similar extent and appeared to involve a similar mechanism as each other. The quality of the diet was more important than quantity for the development of resistance, and soluble carbohydrate was an essential component.     

Effects of molybdenum intake on primary infection and subsequent challenge by the nematode parasite Trichostrongylus colubriformis in weaned Merino lambs.

In a study designed to test the effect of molybdenum (Mo) on resistance to trichostrongylosis, the Mo content of the diet during primary infection of 8-month-old Merino lambs with Trichostrongylus colubriformis was found to affect the number of worm eggs found in the faeces during challenge and the total number of worms in the jejunum after six weeks of challenge infection. The optimal intake of Mo in this experiment was in the range of 4-8 mg sheep(-1)day(-1), approximately equivalent to feeding a diet containing 6-10 mg Mo kg(-1)dry matter (DM) and to 0.15-0.30 mg Mo kg(-1)liveweight. Lambs fed Mo at this rate showed a 90 per cent reduction in faecal egg count and total worm count six weeks after challenge compared with lambs fed quantities of Mo outside this range. The mechanism by which Mo exerted these effects was not defined, although the interactions of molybdenum, worm establishment and faecal egg count suggested that this trace element may be acting via an effect on the host's acquired immune response. This hypothesis is supported by the observed enhancement of immune responses (intestinal antibody and granulocyte numbers and in vitro worm-specific proliferation of lymphocytes) associated with Mo intake of 0.15-0.3 mg kg(-1)Lwt.