Computed tomography and radioactive 32P detected phosphorus impairment in metabolism, reduced bones density and animal performance caused by mixed infection of Haemonchus contortus and Trichostrongylus colubriformis in sheep.

Phosphorus (P) is one of the main minerals present in the animal body and exerts crucial functions in the organism. P is present at all cell membranes and integrates the structure of bones, being necessary its supplementation in ruminants due to the deficiency of this mineral in the pastures. One of the principal factors that compromise its metabolization are gastrointestinal nematodes (GIN). Thus, the objective of this study was evaluate the performance and metabolism of P through its distribution in the animal body, density of bones and muscles, dynamic fluxes, biological availability and half live of P, concentration of P in tissues and bones of lambs simultaneously infected with the most prevalent GIN to sheep, in tropical or subtropical areas, (Haemonchus contortus and Trichostrongylus colubriformis) using the isotopic dilution technique with 32P radioisotope. Twenty Santa Ines sheep with seven months of age and averaging initial weight of 30.8 ± 6.41 kg were used and allocated to one of two treatments. Ten animals were orally infected (a single dose of 30,000 L3 larvae of T. colubriformis + 10,000 L3 larvae of H. contortus), and ten animals were not infected (control group). During the experimental, samples of blood, feces, urine, and diet refusals were collected and weighting were performed. A computed tomography was performed twice, before infection and at the end of the experiment, to evaluate changes in body composition. On 64-d after experimental infection, animals received an intravenous injection of 32P solution, and 7-d after they received radioisotope injection. The experimental animals were slaughtered, and tissue and bones were collected for P concentrations. The results showed that the parasitic infection compromised the absorption of P, impairing the metabolism, decreasing the mineral bioavailability increasing P bones reabsorption, and reducing bones density, also negatively compromising the infected animal performance.

Effects of Acacia mearnsii supplementation on nutrition, parasitological, blood parameters and methane emissions in Santa Inês sheep infected with Trichostrongylus colubriformis and Haemonchus contortus.

Gastrointestinal nematodes are a main problem for ruminant production, reducing animal performance and increasing environmental impact per unit of animal product generated. Tannin supplementation may lead to positive results regarding aspects such as parasitic infections and methane (CH4) emissions. Therefore, the objective of this experiment was to evaluate the effects of the condensed tannins (CT) extract made of powdered Acacia mearnsii bark (PAB) on nutrition, parasitic status and CH4 emissions in sheep artificially infected with Trichostrongylus colubriformis and Haemonchus contortus. Twenty 10-month old Santa Inês lambs (24.7 ± 3.14 kg of initial body weight) were used in a 50-day trial. Animals were divided in four treatment groups according to parasitic infection and PAB supplementation: two control groups without infections, one without PAB (C-) (n = 4) and one with PAB (C+) (n = 4); two infected groups, one without PAB (I-) (n = 6) and another receiving PAB (I+) (n = 6). Initially, animals were kept in individual pens where they were fed ad libitum chopped tifton 85 hay (Cynodon spp.) and 210 g/animal/day of concentrate. On the first day of experiment, animals of I- and I+ groups were artificially infected with infective larvae (L3) of T. colubriformis and H. contortus. Lambs were weighed fortnightly to calculate average daily body weight gain (ADG). Blood and faeces samples were also collected in the same moment of weighing for the evaluation of blood parameters and faecal egg count (FEC) respectively. After 40 days of experiment, measurements of CH4 emissions in small chamber system started and following that, apparent total tract digestibility (ATTD) assay was carried out in metabolic cages. In the end of experimental period (50 days), lambs were slaughtered and samples of abomasum and small intestine content were collected for worm count, identification, and eggs/female count. No significant (p > 0.05) treatment effects were verified for ADG, ATTD and worm count. Blood parameters were affected in both infected groups (p < 0.05) from the 28th experimental day onwards, when these animals started to show reduced red blood cells, haemoglobin and packed cell volume when compared to C- and C+. Decreased FEC was verified in I+ when compared to I- and also, H. contortus eggs/female worm for I+ was lower than for I- (p < 0.05). Both infected groups showed higher CH4 emissions than the control groups (p < 0.05). Results highlighted the anthelmintic potential of PAB and indicated methanogenic effect of parasitic nematode infections.

Condensed Tannin Changes along the Digestive Tract in Lambs Fed with Sainfoin Pellets or Hazelnut Skins.

The variable anthelmintic efficacy of condensed tannins (CT) against gastrointestinal nematodes may depend on CT concentration, composition, or fate along the digestive tract. We analyzed CT concentration and composition by acetone-HCl-butanol and thiolysis coupled to HPLC-MS in digesta and feces of lambs. Lambs had been infected with Haemonchus contortus and Trichostrongylus colubriformis and received sainfoin pellets and hazelnut skins of contrasting prodelphinidin/procyanidin ratios. The digesta and feces had lower CT concentrations than the original feeds but similar concentration patterns across the digestive compartments. The changes in assayable CT concentrations between rumen, abomasum, and small intestine may be due to complex formation between CT and other dietary components. However, the large CT disappearance (61-85%) from feed to feces could also indicate that CT may have been structurally modified, degraded, or absorbed during digestion. Interestingly, there were no changes in the structural features of assayable CT in the digesta.

ANTHELMINTIC EFFECTS OF DRIED GROUND BANANA PLANT LEAVES (MUSA SPP.) FED TO SHEEP ARTIFICIALLY INFECTED WITH HAEMONCHUS CONTORTUS AND TRICHOSTRONGYLUS COLUBRIFORMIS.

BACKGROUND: Helminths is a endoparasites that cause the major losses for profitable sheep production in Brazil. The increased development of resistant strains of endoparasites have enforced the search for sustainable alternatives. The aim of this paper was to provide information about endoparasites control with banana leaves in infected sheep as alternative control strategies and see its viability. MATERIALS AND METHODS: In this study, we performed two trials to investigate the anthelmintic properties of banana leaves on endoparasites in sheep. In Trial 1, twelve sheep were artificially infected with Trichostrongylus colubriformis; in Trial 2, eleven sheep were artificially infected with Haemonchus contortus. Clinical examinations, packed cell volume, total protein, faecal egg counts (FECs) and egg hatchability tests (EHTs) were performed. At the end of the trials, the sheep were humanely slaughtered, and total worm counts were performed. RESULTS: In Trial 1 and 2, no significant FEC decreases were note but significant diference in EHTs were observed. Total worm counts, clinical and haematological parameters did not reveal significant changes between the treatment and control groups. These results suggest that feeding dried ground banana plant leaves to sheep may reduce the viability of Trichostrongylus colubriformis eggs, and this anthelmintic activity is potentially exploitable as part of an integrated parasite management programme. CONCLUSION: However, further investigation is needed to establish the optimal dosage, develop a convenient delivery form and confirm the economic feasibility of using banana plantation byproducts as feed for ruminant species. Abbreviations: Coproculture test (CT)., Faecal egg count (FEC)., Egg hatchability test (EHT).

Dried, ground banana plant leaves (Musa spp.) for the control of Haemonchus contortus and Trichostrongylus colubriformis infections in sheep.

To evaluate the anthelmintic effect of Musa spp. leaves, 12 animals were artificially infected with Haemonchus contortus, and another 12 animals were infected with Trichostrongylus colubriformis. Then, both treatment groups were offered 400 g of dried ground banana plant leaves, and the control animals were offered only 1000 g of coast cross hay. During the trials, the animals received weekly physical examinations. The methods used to evaluate the efficiency of this treatment were packed cell volume, total plasma protein and faecal egg counts, and egg hatchability tests were performed on days -2, +3, +6, +9, +13 and +15. Coproculture tests were performed on day -2 to confirm monospecific infections. In the FEC and EHT, a statistically significant difference (0.04, 0.005; p < 0.05) was noted for T. colubriformis. There were no statistically significant differences (p > 0.05) for Haemochus contortus group in all tests. Our results confirmed previous findings suggesting that dried ground banana plant leaves possess anthelmintic activity.

Anthelmintic effect of carob pods and sainfoin hay when fed to lambs after experimental trickle infections with Haemonchus contortus and Trichostrongylus colubriformis.

The aim of the study was to compare the in vivo anthelmintic activity of sainfoin hay (Onobrychis viciifolia) and carob pod meal (Ceratonia siliqua) against gastrointestinal nematodes. Seven days before infection, 64 naive lambs were assigned to four different groups: Group S received sainfoin hay and group CAR was fed with carob pods. The remaining lambs received lucerne hay (Medicago sativa) and were assigned to positive (non-treated, NT) and negative (treated, T) control groups (treatment with albendazole). On day 0, lambs were artificially trickle infected for 6 weeks, with a mixture of infective larvae of Haemonchus contortus and Trichostrongylus colubriformis. Parasitological and pathophysiological parameters were measured repeatedly during the 2-month study. Compared to the NT group, decreases in egg excretion were observed in the CAR and S groups with significant differences only found for sainfoin (p < 0.05). At necropsy, group S showed decreases in the total worm numbers of both nematode species with significant differences for H. contortus. In contrast, no differences were noticed for the CAR group. Compared to the NT group, lower values for fecundity of female H. contortus were found in the S and CAR groups, however differences were non-significant. No differences in body weight gains were found between groups. Consistent results were found showing significantly higher packed cell volume (PCV) values in the T and S groups compared to NT and CAR groups. Overall, these results confirm a positive effect associated with the feeding of lambs with tanniniferous resources on host resilience (PCV values) and against gastrointestinal parasitic nematodes by affecting some biological traits of worm populations (e.g. eggs per gram of faeces and worm numbers). However, the anthelmintic effects differed between the two tannin-containing resources, which might be associated with the quantity and/or quality of secondary metabolites (condensed tannins and/or other polyphenols).

In vitro ovicidal and larvicidal activity of methanolic leaf extract of Manihot esculenta (cassava) on susceptible and resistant strains of Trichostrongylus colubriformis.

This study aimed to represent the first report of the ovicidal and larvicidal activity of the methanolic leaf extract of Manihot esculenta (cassava) against eggs and larvae of susceptible and resistant strains of Trichostrongylus colubriformis. As well as, to determine the total tannin compounds, antioxidant activity and toxicity of the extract. The egg hatch test was used to evaluate ovicidal activity against unembryonated eggs, whereas larval feeding inhibition assay and MTT-formazan assay were used to evaluate larvicidal activity against first (L(1)) and infective (L(3)) larvae, respectively. The results showed no significant differences were detected between the sensitivities of susceptible and resistant strains of T. colubriformis to the extract. Eggs, L(1) and L(3) were significantly affected (P<0.001) compared with negative control, and L(1) were more sensitive than the eggs and L(3). The total tannin compounds were investigated using tannin quantification assay and determined by 254.44 TAE/mg. The antioxidant activity was evaluated using the DPPH radical scavenging assay and the median inhibition concentration (IC(50)) was determined by 2.638 mg/ml. Acute oral toxicity at dose of 5,000 mg/kg, and sub-chronic oral toxicity at 500 and 1,000 mg/kg of the extract were observed in male and female Sprague-Dawley (SD) rats. The acute oral toxicity revealed that the median lethal dose (LD(50)) of methanolic extract of cassava leaves on SD rats was greater than 5,000 mg/kg, whereas the sub-chronic oral toxicity did not show observed adverse effects at 500 and 1,000 mg/kg per day for 28 days. In conclusion, the methanolic extract of cassava leaves has direct ovicidal and larvicidal activity against T. colubriformis strains with a safety margin for animals, and it may be potentially utilized as a source of natural antioxidants.

Anthelmintic efficacy of five tropical native Australian plants against Haemonchus contortus and Trichostrongylus colubriformis in experimentally infected goats (Capra hircus).

The study of the anthelmintic properties of plants rich in plant secondary metabolites can provide ecologically sound methods for the treatment of parasites on grazing animals. The purpose of the present study was to evaluate the anthelmintic effect of five tropical native Australian plant species rich in plant secondary metabolites on adult Haemonchus contortus and Trichostrongylus colubriformis in experimentally infected goats. Thirty young, nematode-free goats were infected with 2500 H. contortus and 5000 T. colubriformis infective larvae thrice weekly for a week (day 1-7 of the experiment). On day 27 after first infection, the goats were allocated into six groups of five animals per group. From day 28 to day 35, fresh leaves from Acacia salicina, Acacia nilotica, Eucalyptus corymbia, Casuarina cunninghamiana and Eucalyptus drepanophylla were included in the goats diet. Five groups were offered leaves from one of these plant species and one group, the untreated control, received only the basal diet formulated with 20% Medicago sativa and 80% Avena sativa. Following plant material administration, the goats were monitored daily until day 40 and then slaughtered on day 41. Total faecal worm egg output, total production of larvae recovered from faecal cultures, total post-mortem worm burdens and the per capita fecundity of female worms were estimated. The toxicity of the plant species for the goats was measured by histopathological analyses of liver and kidney samples. Results showed that goats feeding on the plant material rich in plant secondary metabolites had significantly lower egg output compared to the control goats (P<0.05). A similar response was found for larval production in both H. contortus and T. colubriformis supporting that egg output was affected in both species. Although the total worm burdens were not affected by the plant material (P>0.05), the per capita fecundity was significantly reduced by E. corymbia, A. nilotica and A. salicina (P<0.05). No signs of toxicity were detected in the liver or kidney samples. It is concluded that goats can benefit from the short-term ingestion of plant secondary metabolites, which reduce the total faecal egg output and thus decrease the potential for re-infection from the pasture.

Anthelmintic and cytotoxic activities of extracts of Markhamia obtusifolia Sprague (Bignoniaceae).

The anthelmintic activity of Markhamia obtusifolia Sprague (Bignoniaceae) leaf extracts was evaluated against the ruminant gastrointestinal nematode parasite Trichostrongylus colubriformis (Nematoda: Strongylida) using the in vitro egg hatch test. Also, the cytotoxic activity of aqueous extracts of M. obtusifolia was evaluated in cell line cytotoxicity assays. The results indicated that the effective concentration (EC(50)) for the water extract of M. obtusifolia leaves (0.46 mg/mL; Confidence Interval [CI] 0.3-0.5mg/mL) was significantly lower than the EC(50) for the acetone extract of M. obtusifolia (0.8 mg/mL; CI 0.7-1mg/mL). Aqueous extracts were twice as potent as the acetone extracts. The EC(90) (0.2mg/mL; CI 0.1-0.02) for thiabendazole (positive control) was significantly lower than the EC(90) for the water extract of M. obtusifolia (10.7 mg/mL; CI 8.3-13.7 mg/mL). In the cytotoxicity bioassay, the lethal concentration (LC(50)) for the aqueous extract of M. obtusifolia was 0.476 mg/mL, which was relatively high (low toxicity) in comparison to the highly toxic berberine (LC(50)=9.80 µg/mL). The current study showed that M. obtusifolia plant extracts possess anthelmintic activity and are relatively non-cytotoxic, thus providing support for their use in traditional veterinary practices.

Animal grazing selectivity and plant chemistry issues impact on the potential of Rhagodia preissii as an anthelmintic shrub.

Rhagodia preissii had shown significant in vitro anthelmintic activity in a previous study, we examined the effect of including this shrub in the diet of sheep infected with Trichostrongylus colubriformis. Worm-infected merino wethers were grazed for 7 weeks on either R. preissii or annual pasture, and faecal egg counts (FECs) were conducted weekly. Plant material was collected weekly from eaten and uneaten plants, and analysed for levels of plant secondary metabolites (tannins, oxalates, saponins) and in vitro anthelmintic activity. While mean FECs were consistently lower in sheep grazing R. preissii compared to pasture (reductions of 20-74%), the differences were not significant. There was no relationship between grazing preference (eaten or uneaten) and in vitro anthelmintic activity of plant extracts. The levels of saponins and oxalates did not correlate with grazing preference or in vitro anthelmintic activity, while tannins were not responsible for the anthelmintic activity. While the identity of the grazing deterrent and in vitro anthelmintic compounds remain unknown, the presence of plants which were both highly preferred by the sheep and showed in vitro anthelmintic activity indicates a potential to develop the species as an anthelmintic shrub through selection of shrub populations dominated by such plants.

In vitro activity of Peltophorum africanum Sond. (Fabaceae) extracts on the egg hatching and larval development of the parasitic nematode Trichostrongylus colubriformis.

Trichostrongylus colubriformis is an important cause of parasitic gastroenteritis in ruminants, where it causes protracted diarrhoea, rapid loss of weight, loss of production and death. The in vitro efficacy of extracts of Peltophorum africanum was determined against this parasitic nematode. Eggs and larvae of T. colubriformis were incubated at 23 degrees C in the extracts of the leaf, bark and root of P. africanum at concentrations of 0.008-25 mg ml-1 for 2 and 5 days, respectively. Thiabendazole and water were used as positive and negative controls, respectively. Inhibition of egg hatching and larval development increased significantly (P<0.05) with increasing concentrations of the extracts. Concentrations of 0.2-1.0 mg ml-1 of the extracts of leaf, stem bark, and root bark of P. africanum completely inhibited the hatching of eggs and development of larvae. No eggs and larvae of T. colubriformis could be observed in wells incubated with all the three extracts at concentrations of 5 and 25 mg ml-1. The in vitro model results support the traditional use of P. africanum against nematode parasites. Further research is required to isolate and structurally identify the active anthelmintic compounds, and to improve methods of plant extraction of the effective anthelmintic components that will be readily adaptable for use by rural communities against helminthosis.

Green tea flavan-3-ols and oligomeric proanthocyanidins inhibit the motility of infective larvae of Teladorsagia circumcincta and Trichostrongylus colubriformis in vitro.

The effects of a hot water infusion and an aqueous acetone extract of green tea (Camellia sinensis) on the motility of infective larvae of the sheep nematodes Teladorsagia circumcincta and Trichostrongylus colubriformis were investigated under in vitro conditions. The infusion and extract dose-dependently inactivated the infective larvae as assessed by the larval migration inhibition (LMI) assay. To determine the components responsible for the inhibitory activity, the hot water infusion and aqueous acetone extract of green tea were fractionated on Sephadex LH-20 and the green tea extract fractions (GTE-I-VIII) characterised by mass spectrometry. The larvae were exposed to increasing concentrations of these GTE fractions. Fractions containing epigallocatechin gallate (EGCG) and proanthocyanidin oligomers were most effective. GTE fractions were more effective against T. circumcincta than T. colubriformis larvae as assessed by the LMI assay.

The effect of feeding sulla (Hedysarum coronarium) or lucerne (Medicago sativa) on lamb parasite burdens and development of immunity to gastrointestinal nematodes.

Two experiments investigated the efficacy of the legume Hedysarum coronarium (sulla), which contains condensed tannins (CTs), for reducing gastrointestinal nematode infections relative to lucerne. Experiment 1 was aimed to show whether the lower faecal egg count (FEC) and larval establishment previously reported in lambs grazing sulla were due to direct effects of the forage on Ostertagia circumcincta and Trichostrongylus colubriformis or were mediated through an enhanced immune response. Experiment 2 evaluated the impact of feeding sulla relative to feeding lucerne (Medicago sativa), before, at, or after larval challenge on subsequent FECs and nematode burdens. In experiment 1, 64 Romney lambs were fed either freshly cut lucerne or sulla (32 lambs per herbage) for the duration of the trial. Within each herbage there were four treatment groups (n=8 per group). Initial levels of immunity were assessed in uninfected (UN) lambs which were maintained parasite-free until challenged with 15,000 O. circumcincta and 15,000 T. colubriformis larvae on day 63, and slaughtered on day 81. The other three treatment groups were trickle-infected with each of 5000 O. circumcincta and 5000 T. colubriformis larvae three times per week from day 1 to 35. Non-steroid infected (CONTROL) and steroid-treated (STER) groups were treated with anthelmintic on day 49 and challenged with 15,000 O. circumcincta and 15,000 T. colubriformis on day 63 and slaughtered on day 81. The STER lambs were given dexamethasone trimethylacetate from day 49 to 81 to determine effects of immunity on parasite infection. From day 35 an establishment group (EST) on each herbage was fed a common pelleted lucerne diet and slaughtered on day 56 to determine nematode establishment during trickle-infection. Diet did not affect FECs but feeding lucerne increased (P<0.05) numbers of T. colubriformis in CONTROL lambs compared to those fed sulla. O. circumcincta numbers were lower (P<0.05) in UN lambs fed sulla than lucerne. The sulla diet was associated with higher (P<0.05) antibody titres against secretory-excretory antigens to adult O. circumcincta and to adult and larval T. colubriformis, so there appeared to be some immunogenic response to the sulla diet but effects upon T. colubriformis numbers were not significant. The second experiment involved 48 Romney lambs grazing conventional pasture which were infected with 10,000 each of O. circumcincta and T. colubriformis larvae either 7 days before, 7 days after, or at the time they commenced grazing either sulla or lucerne. Lambs which grazed sulla had lower (P<0.05) FEC and lower (P<0.05) O. circumcincta burdens than lambs which grazed lucerne but timing of infection had no effect on FEC or worm burdens. T. colubriformis numbers were not affected by treatment or herbage. In conclusion, the sulla diet resulted in lower O. circumcincta numbers compared to lucerne outdoors and some evidence of an immunogenic response was obtained indoors. However, neither the herbage nor the immunogenic response reduced T. colubriformis numbers in either experiment.

The effects of condensed tannins supplementation of foods with different protein content on parasitism, food intake and performance of sheep infected with Trichostrongylus colubriformis.

The aims of the present study were to investigate (1), the potential anthelmintic properties and (2), the nutritional consequences of commercially available condensed tannins on parasitised sheep fed, ad libitum, either a high- or a low-protein food. For this purpose, forty-eight previously parasite-naïve sheep (n 12) were infected with 2000 Trichostrongylus colubriformis larvae/d for a 67-d experimental period. Two experimental foods were made: a low (L), formulated to be inadequate in meeting the requirements of growing sheep for metabolisable protein (MP), and based on wheat, citrus pulp, and oatfeed; a high (H), expected to be above the requirements of growing sheep for MP, based on similar ingredients but supplemented with protected soyabean meal. Two additional foods were made by adding 60 g Quebracho (a condensed tannins (CT) extract)/kg fresh matter to foods L and H (foods LQ and HQ respectively). This level of Quebracho supplementation has been previously shown to reduce the level of parasitism in restrictedly fed, parasitised sheep. The experiment was divided into two periods: period 1 (P1, day 1-38) and period 2 (P2, day 39-67), each one associated with different phases of an intestinal parasitic infection. Six sheep from each group were slaughtered at the end of P1, and the remaining sheep were slaughtered at the end of P2 (day 67). Although faecal egg counts (FEC; number of parasite eggs/g faeces) and total egg output were reduced in sheep offered the supplemented foods during P1 (P<0.05), worm burdens on day 38 were unaltered. Neither Quebracho supplementation nor food protein content during P2 affected FEC and worm burdens. Food intake and performance were higher in sheep offered food HQ compared with sheep offered food H (P<0.05); no differences were observed in sheep offered foods LQ and L throughout the experiment. The previously shown anthelmintic properties of CT were not observed following ad libitum intake of either low- or high-protein foods supplemented with Quebracho extract. Higher levels of CT supplementation may be required to reduce parasitism and consequently improve the performance of parasitised sheep, when fed ad libitum. Supplementation with CT conferred advantages on the performance of parasitised sheep on a high- but not on a low-protein food.

The effects of urea supplementation on production and parasitological responses of sheep infected with Haemonchus contortus and Trichostrongylus colubriformis.

Merino wether lambs were individually confined and fed a basal diet of oaten chaff containing essential minerals which was untreated or contained 3% urea. Within each dietary group animals were orally infected with either 200 H. contortus (H), 1000 T. colubriformis (T) or both species (H + T) thrice weekly or remained uninfected (C). Weight gain, wool production, and parasite burden were measured over a 19-week period. Sheep on the diet containing urea gained more weight, consumed more feed and grew more wool of higher fibre diameter than their counterparts given no urea. On both diets uninfected sheep consumed more feed than infected sheep and the sheep given no urea and infected with both H and T worm species consumed the least feed. Parasitised sheep gained less weight than uninfected control sheep. Sheep with urea in their diet had lower faecal egg counts when infected with H alone or with H and T but there was no effect of urea on egg count of sheep infected with T alone. In contrast, T numbers after slaughter were reduced in sheep fed diets containing urea whereas H numbers were not affected by diet. It was concluded that supplementation with urea can increase resilience to parasitism thereby improving production and also enhance resistance mechanisms against worms in young sheep on low quality roughage diets. These responses can be partly attributed to stimulation of feed intake, presumably due to enhanced ruminal digestion, but also to elevated rumen NH3-N levels which would be expected to have increased rumen microbial protein synthesis and availability to the intestines.

Establishment and fecundity of Ostertagia circumcincta and Trichostrongylus colubriformis in lambs fed lotus (Lotus pedunculatus) or perennial ryegrass.

Forty four 12-14 week old Polled Dorset ram lambs, raised helminth free from birth, were used to investigate the effects of condensed tannins (CT) in lotus (Lotus pedunculatus) on lamb growth and gastrointestinal nematode establishment and fecundity. Condensed tannins bind to proteins in the rumen and increase the flow of protein to the intestines. Lambs were allocated to either a ryegrass (Lolium perenne) or lotus diet fed ad libitum, with or without twice daily drenching of polyethylene glycol (PEG) which binds with and deactivates the CT. One week after allocation to the diets, each lamb was infected per os with 10,000 Ostertagia circumcincta and 10,000 Trichostrongylus colubriformis larvae and slaughtered 28 or 29 days later. Lambs fed lotus had lower faecal egg counts (P < 0.0001) and lower O. circumcincta burdens (P < 0.001), fewer female O. circumcincta (P < 0.0001) and higher faecal dry matter (P < 0.001) than lambs fed ryegrass, but numbers of T. colubriformis nematodes were not affected. This trial did not ascribe beneficial effects of L. pedunculatus to CT per se and the use of PEG does not seem appropriate to studies of parasitology.

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.

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.

Use of guniea pigs to assay anthelmintic resistance in ovine isolates of Trichostrongylus colubriformis.

Guinea pigs infected with Trichostrongylus colubriformis were used to develop an assay for anthelmintic resistance by determination of worm burdens following treatment with test anthelmintics. To achieve comparable efficacy with the recommended dose of thiabendazole and levamisole in sheep, dose rates in guinea pigs had to be increased two to four fold. For example, thiabendazole at 100 mg/kg in guinea pigs was 96.6 per cent effective against a thiabendazole susceptible (GS) strain of T colubriformis, but had no effect against a thiabendazole-resistant (VRSG) strain. In sheep 50 mg/kg of thiabendazole would have a similar efficacy against each strain respectively. Morantel tartrate at 10 mg/kg in guinea pigs was 99 to 100 per cent effective against the GS strain but only 54 per cent effective against a morantel resistant (PF4) strain. A slope ratio assay was used to calculate the relative potency of anthelmintics by comparing efficacies against resistant strains with efficacy against the GS strain. Resistance of the VRSG strain to thiabendazole was confirmed with a relative potency for this drug of 0.047 in guinea pigs. The PF4 strain was resistant to both thiabendazole and levamisole which had relative potencies of 0.168 and 0.255 respectively. The advantages of this statistical treatment together with the cost and time savings of the guinea pig model over a conventional critical anthelmintic assay in sheep are discussed.

Subclinical trichostrongylosis in growing lambs produced by continuous larval dosing. The effect on performance and certain plasma constituents.

Three groups of eight-four-month-old worm-free lambs were used to investigate the effect of Trichostrongylus colubriformis infection on growth rate, food intake, and concentration of plasma constituents. Groups ALC and ALI were fed ad libitum, the latter being dosed daily with 2500 infective larvae for 14 weeks. Lambs in group PF were paired to individuals in group ALI and offered the same quantity of food as the infected pair consumed. Hypophosphataemia and hypoalbuminaemia developed in ALI at week 3 and hyperglobulinaemia was present from week 6. There was no significant difference in plasma calcium, protein, urea or glucose concentration between the three groups at any stage. No clinical signs of parasitism were seen in seven of eight ALI lambs and egg counts were generally less than 2000 epg. Worm burdens of 1500-82,900 were found at slaughter. Group weight gain was only 50 per cent of PF lambs which received the same digestible energy intake. Group ALI consumed 15-5 kg dry matter per kg gain in bodyweight compared with 8-6 kg/kg and 9-8 kg/kg for ALC and PF respectively. The infections resulted in extensive villous atrophy and flattening of the intestinal mucosa. Bone growth was arrested and osteoporosis present in five group ALI lambs. The possible use of plasma constituents, particularly phosphorus, as aids for the diagnosis of the early stages of trichostrongylosis in the field is assessed.