The development of Trichostrongylus colubriformis larvae on a range of herbage species or on plots of differing topographical aspect.

Five "contaminations", where faeces containing Trichostrongylus colubriformis eggs were deposited on pasture and serially recovered, were used to compare the rate of decline of faecal mass and larval development. In the first three contaminations, faeces from a common source were deposited on swards of browntop (Agrostis capillaris cv Grasslands Muster), ryegrass (Lolium perenne cv Grassland Nui), white clover (Trifolium pratense cv Grassland Tahora), or onto bare ground in the late spring, summer or autumn. The last two contaminations were done on the north facing aspect or south facing aspect of hill country pastures in summer and autumn. Number of free-living nematodes (first- and second-stage larvae (L(1) and L(2)) and soil dwelling nematodes) and third stage larvae (L(3)) recovered from faeces were counted. In spring there was a significant (P<0.01) effect of sward type on the mass of faeces remaining, with greatest mass remaining on browntop and ryegrass 28 days later, and less on bare ground and white clover. In summer there were more (P<0.05) faeces remaining on browntop than on other herbages which had little faeces remaining and which did not differ one from another. In autumn there was a rapid decline in faecal mass. All faeces were gone from white clover and ryegrass swards by day 10 and from browntop and bare ground by day 14. The number of free-living nematodes did not differ markedly between seasons, ranging from 5 to 8.5% of eggs deposited. The number of L(3) recovered was low in spring ( approximately 0.4% of eggs deposited) and did not differ between swards. In summer, more (P<0.05) L(3) were recovered from faeces deposited on swards of ryegrass and white clover than from bare ground or browntop. Most L(3) were recovered from days 7 to 14 ( approximately 1.3% of eggs deposited). In the autumn, low numbers of L(3) were recovered from browntop on day 3 and ryegrass on day 7 (0.2% of eggs deposited) with virtually no L(3) recovered from faeces placed on white clover or bare ground. There were significant (P<0.001) effects of aspect on the amount of faecal mass remaining in both summer and autumn with less faeces remaining on the south facing aspect than on the north. This was particularly evident during the summer when virtually all of the faeces were intact on the north facing aspect but only 40% was remaining on the south on day 28. In the autumn, while faeces were completely gone from both aspects by day 28 but there were less (P<0.05) faeces remaining on the south facing aspect from days 3 to 18 than from the north. There was no aspect effect in either season on the number of free-living nematodes recovered which averaged 8-11% of eggs deposited. In both seasons a greater number of L(3) were recovered from faeces on the south facing aspect than on the north, particularly 3-10 days after faecal deposition. In summer the rise in L(3) recovered in faeces was more rapid on south facing aspect than on the north but both attained a maximum level of approximately 4% of eggs deposited. In autumn on day 3 there was a rapid rise on south facing aspect to approximately 21% of eggs deposited followed by a gradual decline on day 10 while on the north facing aspect numbers of L(3) recovered only attained 10% of eggs deposited.

The effect of diet fed to lambs on subsequent development of Trichostrongylus colubriformis larvae in vitro and on pasture.

Contrasting herbage diets were fed to lambs to evaluate their effect on subsequent development of Trichostrongylus colubriformis larvae in faeces and on pasture. The diets had either no condensed tannin (CT), lucerne (Medicago sativa cv. Otaio), white clover (Trifolium repens cv. Tahora), or had moderate to high concentrations of CT, sulla (Hedysarum coronarium cv. Grassland Aokau), Lotus corniculatus (cv. Grasslands Goldie), L. pedunculatus (cv. Grassland Maku), Dorycnium pentophyllum, and Dorycnium rectum. Trials were carried out in summer (warm) and in autumn (cool and moist). In summer, egg viability was evaluated in vitro with egg hatch and larval development assays. In both seasons faeces were placed on pasture to compare recovery of eggs and larvae from faeces and larvae from herbage on the high and low fertility farmlets on the AgResearch Ballantrae Hill Country Research Station. D. rectum and D. pentophyllum diets decreased (P<0.01) egg hatching and larval development in laboratory assays relative to other diets. In summer, the number of larvae recovered from faeces placed on pasture was far greater (P<0.001) if the lambs had been fed lucerne than any other diet, whereas recovery was always lowest from faeces of sheep fed D. rectum and D. pentophyllum. Although dietary differences were lower in autumn than in summer, larval recoveries were lower (P<0.05) from faeces of lambs fed D. rectum and L. corniculatus than from white clover, lucerne and sulla diets. This study indicates that the diet of the host can have a significant impact on egg hatching and the subsequent development of T. colubriformis larvae in the laboratory and in the field. In particular, D. rectum consistently reduced T. colubriformis development. Effects measured in vitro generally under-estimated effects measured under field conditions.

The effect of pasture species on parasitism and performance of lambs grazing one of three grass-white clover pasture swards.

Lamb parasitism and performance were compared on three pasture treatments: browntop/white clover, Yorkshire fog/white clover, and ryegrass/white clover swards maintained at a sward height of 5cm in each of two years by using continuous variable stocking management. In year 1, lamb performance (both suppressively drenched (SD) and trigger drenched (TD)) was similar on ryegrass and browntop swards but was poorer (P<0.05) on Yorkshire fog swards. In year 2, lamb performance (both SD and TD) was markedly better on ryegrass than on Yorkshire fog or browntop swards. This coincided with an increase in white clover content in the ryegrass/white clover swards and a decrease in white clover content in the browntop/white clover swards. Lamb performance was related to the white clover content of the swards (R2=85.4, P<0.001 and R2=77.5, P<0.001 for SD lambs and TD lambs, respectively). Differences among pasture treatments in faecal egg count (FEC) of TD lambs were greater in year 2 than in year 1 when no significant pasture effects were observed. In year 2, FEC was lowest in lambs which grazed ryegrass/white clover, intermediate in those which grazed Yorkshire fog/white clover and highest in lambs which grazed browntop/white clover, despite herbage allowance (kg herbage mass/kg lamb mass) being greater on the browntop/white clover and Yorkshire fog/white clover swards than the ryegrass/white clover swards. Circulating antibodies to gastrointestinal parasites were measured at the end of year 2. Antibody titres were highest (P<0.05) in the plasma of lambs which grazed ryegrass/white clover swards, and lowest in lambs which grazed browntop/white clover swards due to differences in levels of antibody to (P<0.05) Haemonchus contortus, Ostertagia circumcincta larvae and adults, but not Trichostrongylus colubriformis larvae or adults. These findings indicate that pasture composition, especially, the proportion of white clover, can help alleviate the production losses due to gastrointestinal parasitism. This information can provide a quick and readily adoptable practice for farmers who aim to reduce drench reliance and could become a key component of any integrated parasite control programme.

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.

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.

Effect of topographical aspect and farm system on the population dynamics of Trichostrongylus larvae on a hill pasture.

The population dynamics of Trichostrongylus colubriformis larvae were compared over two years on contrasting topographical aspects north (warm and dry) and south-facing (cool and moist) hill slopes) on paddocks which form part of the 'non-chemical' and conventional' farm systems at the AgResearch Ballantrae Hill Country Station located in a summer-moist region of New Zealand. Sheep faeces containing 50,000 Trichostrongylus eggs were incubated for 4 days at 25 degrees C and then deposited on each of 36 sub-plots in each of 8 plots in a 2 x 2 factorial design in the summer (summer trial) and again in autumn (autumn trial). Pasture was removed to ground level and larvae extracted from six sub-plots from each plot 2, 4, 6, 8, 11 and 14 weeks after contamination in all trials. Larvae were recovered from two strata, 0-5 cm above the soil surface and > 5 cm above the soil surface. Fewer (p < 0.001) larvae were recovered from herbage (47 vs. 118) and residual faeces (28 vs. 246) from the autumn than from the summer trials. This coincided with more rapid (p < 0.001) faecal disappearance in the autumn trials. In the summer trials, fewer (p < 0.003) larvae were recovered from the herbage (101 vs. 182) and residual faeces (140 vs. 352) from plots on the south than the north facing aspect. In the autumn trials there was a rapid (p < 0.0001) faecal disappearance from the south-facing aspect. In the autumn trials there was a non-significant (p < 0.10) trend for fewer larvae to be recovered from the south-facing aspect (2 vs. 54). This also coincided with more rapid faecal disappearance from the south-facing aspect. There was no effect of farm system on the number of larvae recovered. Despite greater (p < 0.0001) numbers of larvae recovered from the bottom stratum of herbage, the density of larvae (L3/kg DM) tended (p < 0.12) to be higher in the top stratum of herbage. It was concluded that season and aspect have a marked effect on the number of larvae recovered from herbage and that this was inversely related to the rate of faecal disappearance.

Effect of plant species on the larvae of gastrointestinal nematodes which parasitise sheep.

Faeces containing Trichostrongylus colubriformis and/or Ostertagia circumcincta eggs were used to provide four contaminations in each of 2 years on plots of browntop, Yorkshire fog, ryegrass, tall fescue, lucerne, chicory, cocksfoot, white clover, and prairie grass and in the second year a mixed sward of ryegrass/white clover. Third stage larvae were recovered from faeces and from four strata of herbage, 0-2.5, 2.5-5, 5-7.5 and > 7.5 cm above the soil surface at 2, 4, 6, 8, 11, and 14 weeks after faeces were deposited on the swards. Herbage species had a significant (P < 0.0001) effect on the number of larvae recovered. Greatest numbers of larvae, as indicated by ranking analysis, were recovered from Yorkshire fog, ryegrass, and cocksfoot and lowest numbers from white clover and lucerne. The difference between herbages in numbers of larvae recovered was due to the "development success", the ability of larvae to develop to the infective stage and migrate on to herbage, rather than "survival", the rate of population decline once on the herbage. Faecal degradation was most rapid from white clover and browntop, intermediate from tall fescue, lucerne, prairie grass, cocksfoot, and ryegrass, and slowest from Yorkshire fog swards. The numbers of larvae recovered from herbages were related (r2 = 0.59, P < 0.05) with the faecal mass remaining. A greater proportion of the total larvae recovered from the herbage was recovered from the bottom stratum of Yorkshire fog and prairie grass than from white clover, with the other herbages intermediate, indicating that larvae had greater difficulty migrating up Yorkshire fog and prairie grass than the other herbage species. In most herbage species, despite more larvae being recovered from the lowest stratum, larval density (L3/kg herbage DM) was highest in the top stratum. This study has demonstrated that herbage species can have a significant impact on the population dynamics and vertical migration of T. colubriformis and O. circumcincta larvae.

Production, faecal egg counts and worm burdens of ewe lambs which grazed six contrasting forages.

Lambs were grazed on monospecific swards of one of six forages, some containing condensed tannins (CT); sulla (Hedysarum coronarium), Lotus corniculatus and L. pedunculatus and some without CT (lucerne (Medicago sativa), plantain (Plantago lanceolata) and a ryegrass/white clover pasture (Lolium perenne/Trifolium repens)) to evaluate effects of forage type on gastrointestinal nematode burdens, faecal egg count as well as liveweight gain (LWG), wool growth and dag formation over a 42-day period. A new allotment of feed was provided at seven-day intervals, and feed was available ad libitum. Total worm burdens in a pre-treatment slaughter group were ca. 22000 of which ca. 5000 were in the abomasum. Within each forage type, 10 of the 25 lambs (NP) were given an anthelmintic drench at 14-day intervals and the remaining 15 lambs (P) were not drenched. Daily gains of the NP lambs were double that of P lambs which grazed either L. corniculatus, lucerne or pasture, but parasitism had a lesser effect on performance of lambs which grazed sulla and L. pedunculatus. Plantain was not palatable and all lambs performed poorly. Highest daily gains in P lambs were for those which grazed sulla (175 g/day) and L. pedunculatus (160 g/day) with total worm burdens of 13100 and 23000 for the respective treatment groups. The other forages resulted in lower daily LWG in P lambs, and performance was not related to either worm burdens or worm species. This experiment has shown that, when sulla is fed, there is a reduction in worm burdens and faecal egg count (FEC), but with Maku lotus which also contains CT, the good level of performance was achieved despite in high worm burdens and FEC. The mechanisms by which these forages enable high levels of productivity in the face of a parasitism appear to differ, but both could be incorporated into forage feeding systems to reduce dependence on anthelmintic drenches.

Infestation of sheep dung by nematophagous fungi and implications for the control of free-living stages of gastro-intestinal nematodes.

A field trial was conducted to assess the rate at which dung becomes infested by fungi which parasitise nematodes (nematophagous fungi) after deposition. Sheep dung was placed on field plots of bare ground, ryegrass (Lolium perenne), browntop (Agrostis capillaris) and white clover (Trifolium repens) in summer (February) and autumn (April), and subsamples were examined at intervals for the presence of nematophagous fungi. Nematophagous fungi occurred in 71% of 129 samples recovered in February and 57% of 58 samples recovered in April. Arthrobotrys oligospora, Monacrosporium candidum and Nematoctonus spp. were the most frequently isolated nematode-trapping fungi in both seasons. The endoparasitic nematophagous fungus Harposporium leptospira also occurred frequently in dung deposited in February, but not April. Fungi entered dung quickly, with 83% and 58% of dung samples containing nematophagous fungi at 3 days after deposition in February and April, respectively. The percentage of dung infested by nematophagous fungi on plots of bare ground, ryegrass, white clover and browntop was 76%, 75%, 61% and 55%, respectively. Results suggest that a number of species of nematophagous fungi are able to enter dung soon after deposition on a variety of types of ground cover.

Controlling internal parasites in grazing ruminants without recourse to anthelmintics: approaches, experiences and prospects.

Interest in ways of raising stock without using anthelmintics has been stimulated by the desire for intensive grazing systems to adopt more sustainable methods of internal parasite control and by potential premiums for "organic" produce. This requires grazing management systems which are practicable and achieve levels of parasite control sufficient to meet realistic production objectives. This paper will summarise 3 years experience of lamb and cattle production without a recourse to anthelmintics on 2 production systems: a mixed cropping, lamb finishing, and cattle rearing unit and an all grass, mixed-stock, hill country unit in which all non-replacement lambs are sold at weaning. On both units parasite control was almost entirely dependent on integrated grazing management of sheep and cattle and strict systems of grazing management had to be rigidly applied. Acceptable productivity could be more readily achieved in sheep than cattle. Rams selected for resistance to nematodes were also used in both sheep flocks. Their influence on lamb production was equivocal. There is a need for more information on factors influencing parasite epidemiology and for consideration of strategies other than alternate grazing. The impact of pasture species on parasite epidemiology needs to be clarified. Current research indicates substantial differences between grasses in terms of parasite burdens acquired and production losses suffered by lambs grazing them. Grazing management may need to vary with pasture species. Also, specialty forage crops, particularly those containing condensed tannins (i.e. Hedysarum coronarium, Lotus pedunculatus and Lotus corniculatus) hold special promise as a means of countering parasite-induced production losses and dagginess. Using biological control of free-living larval stages and vaccination, may in the long term, also prove useful. Developing effective and acceptable systems for raising stock without using anthelmintics presents a considerable challenge, to parasitologists, as well as to plant breeders, agronomists, and farming systems researchers.

Effect of plane of nutrition before and after 200 kilograms of body weight on mammary development of prepubertal Holstein heifers.

Twenty Holstein heifer calves at 138 d of age and 118 kg of BW were assigned randomly to one of four treatments in a factorial design. Heifers were fed for low (0.7 kg/d) or high (> 1.0 kg/d) gain during two periods. Period 1 was from start to 200 kg of BW, and period 2 was from 200 kg of BW to slaughter, which occurred in the middle of the luteal phase following the third estrus. Calves on the high plane of nutrition during period 2 were heavier at the end of trial and gained more BW between puberty and the end of trial. Those calves also had heavier mammary glands, which had more fat and dry fat-free tissue. Heifers that had been raised on the low plane of nutrition throughout the trial had greater concentrations of DNA and RNA but did not have greater protein in the dry fat-free tissue. Low plane of nutrition during either period increased DNA and RNA concentration in the dry fat-free tissue. Total mass of DNA and RNA did not differ among treatments. Protein mass was increased by the high plane of nutrition during period 2. This study indicated that dietary manipulation affects mammary dry fat-free tissue, protein, and adipose mass in the period up to and including puberty.

Red urine from red deer grazed on pure red clover swards.

Twenty-four red deer hinds with their calves were released on to a newly established pure red clover sward and, 2 days later, red staining of the tail, perineum and hocks was observed. This was presumed to be of urinary origin. Observation of micturition showed that when urine was passed, it was a normal straw colour but it turned scarlet-red about 1 hour after exposure to air. Midstream urine remained the normal colour when held under a pure nitrogen atmosphere immediately after micturition, but it turned red when held in air in the dark, suggesting that the colour change was due to an oxidative rather than a photosensitive reaction. All deer grazing red clover were affected but this did not occur in deer grazing ryegrass/white clover swards. No adverse effects were observed in the deer grazing the red clover, and calf growth was significantly higher than on ryegrass/white clover, suggesting that the red urine had no effect on health or productivity. Blood and urine analyses showed no signs of haemolysis, haematuria or haemoglobinuria. Preliminary chemical analyses suggest that the compounds involved are not those found in the urine of sheep grazing oestrogenic clover. The nature of the compounds have yet to be determined.