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The present study used in vitro assays to determine the relative potency of commercial macrocyclic lactone (ML) anthelmintics against larvae of drug-susceptible and drug-resistant Australian isolates of important parasites of sheep and cattle, Haemonchus contortus and Haemonchus placei, respectively. Cattle pour-on products containing abamectin, ivermectin, eprinomectin, doramectin or moxidectin were diluted in DMSO and used in larval development assays. Abamectin was the most potent chemical (lowest IC50 value) towards the drug-susceptible H. contortus Kirby isolate. The abamectin IC50 was approximately 2-fold lower than those for ivermectin, moxidectin, eprinomectin and doramectin. Moxidectin and abamectin were the most potent chemicals towards the resistant H. contortus Wallangra isolate. This isolate showed resistance ratios up to 70-fold towards eprinomectin. The resistance ratio for this species was lowest with moxidectin (ratio of 4.0-fold). Abamectin was also the most potent chemical towards both drug-susceptible (Bremner) and drug-resistant (Dayboro) H. placei isolates. The larval development assay only showed low levels of resistance for the drug-resistant H. placei, with resistance ratios ranging from 1.7 to 2.0 fold for moxidectin and abamectin, up to 3.3-fold for eprinomectin. This study examined the readily-accessible larval life stages of these parasites in in vitro assays, and, hence, the relationship between our findings and relative drug efficacies in vivo remains to be determined. Despite this, the study accords with some evidence from the use of these anthelmintics in the field in demonstrating the potency of moxidectin and abamectin against ML-resistant H. contortus. The study also highlights the usefulness of eprinomectin as a readily-available compound which is a more sensitive marker for ML resistance in in vitro larval development assays than the other commercial ML compounds examined.

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 (P0.05). 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 (P0.05), the per capita fecundity was significantly reduced by E. corymbia, A. nilotica and A. salicina (P0.05). 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.

The environmental impact of Duddingtonia flagrans , a potential biological control agent for nematode parasites, was assessed in an improved pasture. Sheep feces containing D. flagrans and feces without D. flagrans were placed on pasture plots on four different occasions (series): May, August, November 1998, and February 1999. At 0, 1, 2, 4, 8, 16, and 24 weeks following fecal deposition, the persistence and dispersal of the fungus from the site of deposition were estimated. In addition, the impact of fungal presence on the numbers of non-target, free-living soil nematodes and microarthropods was assessed. For all four series, D. flagrans was not found outside the point of fecal deposition indicating that D. flagrans had not spread in the horizontal plane. D. flagrans was detected at the point of fecal deposition while intact fecal material was present and persisted for 8–24 weeks after initial contamination in association with the soil under the point of deposition. Over time, there was generally a decline in the number of positive observations of D. flagrans presence and the rate of this decline appeared to be seasonal. D. flagrans was observed in the top 10 cm layer of soil frequently, in the 10–20 cm soil layer occasionally, and in the 20–30 cm soil layer rarely. It is thought that rainfall and the activity of macroarthropods (in particular dung beetles) and earthworms may have contributed to this downward movement through the soil profile. Numbers of free-living soil nematodes and microarthropods at the point of fecal deposition varied considerably between samples but were not reduced by D. flagrans . There were no negative effects of fungal presence on the presence of other nematode-trapping fungi. The results of this study indicate that there were no detectable negative environmental impacts of D. flagrans use in a typical improved pasture.

A series of feeding trials was conducted with penned sheep harboring Trichostrongylus colubriformis infections. They were offered barley grains supporting the growth of the nematophagous fungus Duddingtonia flagrans. It was shown that as little as 5 g of grain/sheep per day was sufficient to virtually eliminate larval numbers from faecal culture. This effect persisted for the time that the fungal grains were fed, and for up to 2 days following cessation of feeding this material. Macerated fungal grains were also incorporated into a range of feed block formulations. In all these, D. flagrans was found to survive the manufacturing process and resulted in significant reductions in larval numbers in faecal cultures set up during the feeding period to sheep. This was observed even for sheep that showed only modest and irregular block consumption. These studies demonstrate that supplementary feeding or block administration offer potential deployment options for D. flagrans as a means of biological control of nematode parasites of livestock.

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.

Knox, M.R. and Steel, J.W., 1997. Effects of diet and species on the pharmacokinetics of fenbendazole in cattle. Veterinary Research Communications, 21 (1), 37-43. The plasma concentration profiles of fenbendazole (FBZ), FBZ-sulphoxide (OFZ) and FBZ-sulphone were measured following intraruminal administration of FBZ at 7.5 mg/kg bodyweight in Bos taurus and B. indicus cattle offered three different diets: 100% wheaten chaff, 100% lucerne, and a 50:50 mix of these two diets. No differences between the species were apparent except for a longer time to peak plasma concentration for OFZ in the B. taurus steers fed 100% wheaten chaff. Cattle fed wheaten chaff alone gave greater areas under the concentration-time curve and longer persistence for all metabolites than when the same cattle were fed the other diets. It is concluded that the reduced rate of passage of digesta on lower-quality fibrous diets allows greater time for absorption of FBZ and its metabolites from the gut, thereby increasing systemic availability.

Nutritional insufficiency and gastrointestinal nematode parasitism are major constraints to small ruminant production in south-east Asia and the Pacific Islands. Research on the effects of low cost supplements which supply nitrogen and essential minerals on the ability of small ruminants to resist infection is summarised. In controlled pen studies in young Merino sheep offered a low quality roughage diet of oaten chaff and essential minerals, supplementation with urea reduced the effects of parasitic infection by increasing weight gain and wool production and reducing faecal egg output and parasite burden. In Fiji, field studies have shown that supplementation with urea-molasses blocks can result in increased live-weights of lambs at weaning, increased reproduction rates in maiden ewes and reduction in faecal egg output in grazing sheep. Additional benefits were derived from the inclusion of anthelmintic in the blocks in similar groups of sheep particularly during periods of greater susceptibility to parasites. Pen studies with young goats have shown that urea supplements alone gave no production benefits, but when accompanied by 100 g/d of cotton seed meal beneficial responses were observed. It is expected that parasite control in the small ruminant production systems of developing countries in south-east Asia and the Pacific Islands will benefit from the introduction of low cost nitrogen supplements along with anthelmintic therapy delivered strategically by molasses blocks.

Plasma levels of fenbendazole (FBZ) and its sulphoxide (OFZ) and sulphone (FBZ.SO2) metabolites were measured in goats and sheep during low-level administration of FBZ given by intraruminal infusion or formulated into a urea-molasses feed supplement block (UMB). In experiment 1, 6 goats and 6 sheep were offered UMB containing 0.5 g FBZ/kg (MUMB) and individual block consumption was measured daily for 18 days. In experiment 2, some of the same animals (n = 4 for each species) received FBZ by intraruminal infusion at 1, 1.5 and 3 mg/kg liveweight per day for 7 days at each dosage. FBZ, OFZ and FBZ.SO2 levels were determined in plasma collected every 3 days in experiment 1 and on days 4, 5 and 6 of each infusion period in experiment 2. In both experiments, higher equilibrium levels were observed for the three metabolites in sheep than in goats. Significant linear relationships were observed between the daily FBZ dosages and the plasma levels of the three metabolites in both species. The regression coefficients were significantly higher in sheep than in goats for FBZ and OFZ but not for FBZ.SO2, and they were also significantly higher during MUMB administration than during infusion for all three metabolites in both species. FBZ is a suitable anthelmintic for incorporation into a MUMB formulation for use in livestock production systems where responses to molasses urea supplementation have been demonstrated and gastrointestinal parasitism impairs productivity. The results indicate that target dose rates for goats should be 0.75 mg/kg per day compared with 0.5 mg/kg per day for sheep.

Gastrointestinal helminth parasites impact on livestock production systems throughout the world, and the use of anthelmintics to control this problem has lead to the inevitable development of populations of helminths resistant to these treatments. This, coupled with consumer desires for minimal chemical inputs into food and fibre production, has prompted research into non-chemical approaches to helminth control. Scientists of the “Novel Approaches to the Control of Helminth Parasites of Livestock” group met for the 6th time in August 2010 and this paper summarises that meeting. Six scientific sessions addressed current approaches and topics of interest through formal presentations and discussion of issues raised by the contributing authors. Close interaction between researchers and extension specialists during the meeting has contributed to enhanced prospects for field application of research outcomes in the future.

Implantation of trenbolone acetate (TBA) in conjunction with estradiol-17β (E(2)) increases growth, feed conversion efficiency, and carcass leanness in cattle. Our previous study in Brahman steers suggested that the neuropeptide hormone oxytocin (OXT) may be involved in increasing muscle growth after TBA-E(2) treatment. The present study aimed to determine whether OXT mRNA expression in the longissimus muscle (LM) is also up-regulated in TBA-E(2-)implanted wethers as has been found in steers. Real-time quantitative PCR was used to measure the expression of the gene encoding the OXT precursor, three genes with increased expression in the LM muscle of TBA-E(2)-treated steers, MYOD1 (muscle transcription factor), GREB1 (growth regulation by estrogen in breast cancer 1), and WISP2 (Wnt-1 inducible signaling pathway protein 2), and two genes encoding IGF pathway proteins, IGF1, IGFR, in the LM of both untreated and TBA-E(2)-treated wethers. The expression of OXT mRNA in wethers that received the TBA-E(2) treatment was increased ~4.4-fold (P = 0.01). TBA-E(2) treatment also induced a 2.3-fold increase in circulating OXT (P = 0.001). These data, together with the observation that untreated wethers had much higher baseline concentrations of circulating OXT than previously observed in steers, suggest that wethers and steers have quite different OXT hormone systems. TBA-E(2) treatment had no effect on the expression of IGF1, IGFR, and the muscle regulatory gene MYOD1 mRNA levels in wethers (P ≥ 0.15), but there was an increase in the expression of the two growth-related genes, GREB1 (P = 0.001) and WISP2 (P = 0.04). Both genes are common gene targets for both the estrogen and androgen signaling pathways. Consequently, their actions may contribute to the positive interaction between TBA and E(2) on additive improvements on muscle growth.

We have previously reported marked reductions in faecal worm egg counts (WECs) and drenching frequency in sheep on an intensive rotational grazing system (IRG) in a cool temperate environment with summer-dominant rainfall. These experiments were designed to determine the role of the host and environmental factors in mediating this. The role of host factors was investigated by administering a fixed larval challenge in each of the 4 seasons of the year to groups of 20 young sheep on three different management systems, including IRG. This comprised a mixed larval challenge containing infective larvae of Haemonchus contortus and Trichostrongylus colubriformis 7 days after short-acting anthelmintic treatment. A range of measurements was then made up to day 35 post-challenge. The role of environmental factors was determined by assessing pasture infectivity in four seasons using faecal worm egg counts (WECs) and pooled faecal culture of worm-free tracer sheep. The management systems were high input (HI) with high fertiliser inputs high stocking rate and relatively long grazing periods; typical New England management system (TYP) with moderate fertiliser inputs and stocking rate and relatively long grazing periods and; Intensive rotational grazing (IRG) with moderate fertiliser inputs and stocking rate but very short (mean 5 days) grazing periods and long (mean 103 days) rest periods. IRG sheep had higher mean WEC at 28 and 35 days after fixed larval challenge than HI and TYP sheep in spring (IRG: 9500 ± 1000; HI: 4000 ± 1000; TYP: 7200 ± 1000 eggs/g, P0.01) and summer (IRG: 8400 ± 750; HI: 5300 ± 800; TYP: 4400 ± 700 eggs/g; P0.001) and also had lower live weights during these seasons. There was no difference in WEC after the autumn challenge (IRG: 5100 ± 450 HI: 4500 ± 450; TYP: 4200 ± 450 eggs/g; P ≈ 0.36) but IRG had lower WEC than TYP following the winter challenge (IRG: 2900 ± 400; HI: 2300 ± 400; TYP: 4300 ± 400 eggs/g, P0.01). The tracer sheep (used to determine pasture infectivity) on IRG had significantly lower WECs during winter, spring and summer than those under the other management systems. Faecal culture and larval differentiation revealed that faeces from tracers on IRG contained significantly lower proportions of H. contortus and significantly higher proportions of Trichostrongylus spp. and Teladorsagia circumcincta than faeces from tracers on the HI and TYP treatments. Thus, when IRG was most efficacious for worm control, during spring and summer when short graze and long rest periods were maintained, sheep on this system exhibited greater susceptibility to larval challenge while tracer sheep indicated lower pasture infectivity. This demonstrates that the effects of IRG on WEC are mediated by reduced larval challenge rather than increased host resistance to infection.

We believe this study is the first to consider the genetic and phenotypic divergence between isolates of Haemonchus contortus in Australia. Microsatellite markers have been used to investigate genetic divergence, whilst phenotypic divergence has been considered through individual worm morphology, isolate life history traits and the effect of isolates upon the host. The results are discussed in the context of the likely introduction of H. contortus to Australia, its recent isolation, and the characteristics of sheep and goat farming which might act to either isolate or distribute parasites. We conclude that there is significant observable genetic divergence between isolates of H. contortus in Australia. The divergence may have been under-estimated in this study due to a variety of factors. Phenotypic divergence is also observed, and potentially has significant implications for both economic losses due to haemonchosis on individual properties and for decisions regarding the regulation of stock movements in Australia.

Periparturient Merino ewes obtained from lines of sheep that had been selected either for increased resistance to Haemonchus contortus (R) or at random (C) were supplemented, while grazing at pasture, with either nil or 250 g/day cottonseed meal (CSM) for the 6 weeks prior to or the 6 weeks after the start of parturition. Ewes from both supplement groups had lower (mean 66% reduction) faecal egg counts (FECs) during the postpartum period and this coincided with a period of maternal body weight loss. Factors which increased the rate of maternal body weight loss, such as pregnancy and lactation status, also increased FEC. Evidence is presented that the magnitude of the periparturient rise (PPR) in FEC in grazing ewes will be greatest during periods of maternal weight loss and at these times supplementation to increase metabolisable protein (MP) supply will be most effective in increasing resistance to nematode parasites. The resistance of R ewes to nematode parasites was greater than that of C ewes throughout the experiment and was sufficiently low such that anthelmintic treatment in a commercial environment may not have been required. Irrespective of actual FEC, ewes from all treatment combinations exhibited a PPR in FEC. Reduced FEC of R ewes resulted in reduced apparent pasture larval contamination after 18 weeks of continuous grazing but supplementation was ineffective in this regard. It is suggested that integrated parasite management (IPM) programs for periparturient ewes should make use of both protein supplementation and genetic selection to increase worm resistance and reduce dependency on anthelmintics for worm control.

The effectiveness of protein supplementation and genetic selection to enhance the resistance of periparturient Merino ewes to infection from gastrointestinal parasites was tested in a replicated grazing experiment. One hundred and twenty ewes from lines selected for increased resistance (R) to Haemonchus contortus or at random (C) were subjected to one of the three supplement groups that provided 0 or 250 g per day cottonseed meal for 5 weeks prior to, or for 6 weeks after the start of lambing. Faecal egg counts (FEC) of R ewes were consistently lower than those of C ewes but both groups exhibited a periparturient rise in FEC. Supplementation during the pre-partum period reduced FEC and increased ewe body weight gain. The benefits of pre-partum supplementation in reducing FEC continued to be apparent up to 10 weeks after supplementation ceased. There was a strong suggestion that the benefits to parasite resistance from protein supplementation were greatest in C ewes. Wool growth rates (15%) and birth weights (5%) were greater for C ewes but differences between the lines for lamb body weight had disappeared by day 97. The greatest benefit to resistance from protein supplementation was observed when ewes were experiencing a loss of maternal body weight. Conversely, no benefits to resistance were observed when ewes had moderate (78-107 g per day) rates of maternal weight gain. These results suggest that increased resistance as a result of protein supplementation is dependent on the prevailing supply and demand for scarce nutrients such as metabolisable protein (MP). Both genetic selection and protein nutrition are effective strategies to enhance host resistance to nematode infection during the periparturient period.

A field study was undertaken to determine the effects of feeding Duddingtonia flagrans to young Merino sheep on pasture. A total of 60 mixed sex lambs 4-5 months old were divided into six even groups on the basis of liveweight. On Monday to Friday, each week for 6 months, three groups were offered barley grains on which D. flagrans had been cultured while the other three groups remained untreated. Every 4 weeks liveweights were recorded and faecal samples collected for nematode egg count estimation. Feeding D. flagrans reduced faecal egg counts and tended to improve liveweight gains, but considerable differences were observed between groups within treatment. These differences are thought to result from variations between the groups in consumption of the treated barley with the "best" consumers showing the greater effects of treatment.

Control of helminth infections is a major task in livestock production to prevent health constraints and economic losses. However, resistance to established anthelmintic substances already impedes effective anthelmintic treatment in many regions worldwide. Thus, there is an obvious need for sensitive and reliable methods to assess the resistance status of at least the most important nematode populations. Several single nucleotide polymorphisms (SNPs) in the beta-tubulin isotype 1 gene of various nematodes correlate with resistance to benzimidazoles (BZ), a major anthelmintic class. Here we describe the full-length beta-tubulin isotype 1 and 2 and alpha-tubulin coding sequences of the cattle nematode Ostertagia ostertagi. Additionally, the Cooperia oncophora alpha-tubulin coding sequence was identified. Phylogenetic maximum-likelihood analysis revealed that both isotype 1 and 2 are orthologs to the Caenorhabditis elegans ben-1 gene which is also associated with BZ resistance upon mutation. In contrast, a Trichuris trichiura cDNA, postulated to be beta-tubulin isotype 1 involved in BZ resistance in this human parasite, turned out to be closely related to C. elegans beta-tubulins tbb-4 and mec-7 and would therefore represent the first non-ben-1-like beta-tubulin to be under selection through treatment with BZs. A pyrosequencing assay was established to detect BZ resistance associated SNPs in beta-tubulin isotype 1 codons 167, 198 and 200 of C. oncophora and O. ostertagi. PCR-fragments representing either of the two alleles were combined in defined ratios to evaluate the pyrosequencing assay. The correlation between the given and the measured allele frequencies of the respective SNPs was very high. Subsequently laboratory isolates and field populations with known resistance status were analyzed. With the exception of codon 167 in Cooperia, increases of resistance associated alleles were detected for all codons in at least one of the phenotypically resistant population. Pyrosequencing provides a fast, inexpensive and sensitive alternative to conventional resistance detection methods.

Managing infections of sheep with gastrointestinal nematode parasites (worms) and problems of resistance to anthelmintic treatments continue to be major challenges for graziers on the Northern Tablelands of New South Wales, Australia. The whole-farmlet study of grazing enterprises undertaken by the Cicerone Project tested the broad hypotheses that compared with typical management (farmlet B), internal parasites can be more effectively managed with improved nutrition (farmlet A) or by intensive rotational grazing (farmlet C). Further aims were to identify the major sources of variation in faecal worm egg count (WEC) over the 6-year period and to examine the efficacy of the various anthelmintic treatments used during the experiment. This paper describes the management of sheep worms at the whole-farmlet level during the experiment, and analyses data from the routine WEC monitoring (5644 records) and larval differentiation tests (322 records) carried out on behalf of the Cicerone Management Board and by a doctoral candidate. It complements more detailed investigations published elsewhere. Over the period from July 2000 to December 2006, worm infections in ewes, lambs, hoggets and wethers were, with some exceptions, successfully controlled on the farmlets through a combination of regular monitoring of WEC, treatment with a wide array of anthelmintics and grazing management. Farmlet C had lower mean WEC (444 epg) and annual anthelmintic treatment frequency (3.1 treatments/year) over the whole experimental period than farmlets B (1122 epg, 4.3 treatments/year) or A (1374 epg, 4.7 treatments/year). The main factors influencing WEC were the time since the last anthelmintic treatment, and the anthelmintic used at that treatment. The magnitude of these effects dwarfed those of climatic and management factors that might be expected to influence the epidemiology of gastrointestinal nematode infections via environmental or host-mediated mechanisms. Nevertheless management factors associated with stocking rate and grazed proportion (proportion of each farmlet grazed at any one time), and climatic indicators of both temperature and moisture availability had significant effects on WEC. The results show that, in a region with Haemonchus contortus as the major sheep nematode, improved host nutrition in a higher input system (farmlet A) did not provide more effective control of gastrointestinal nematodes than typical management (farmlet B); however, it was observed that gastrointestinal nematode control was no worse on farmlet A than on farmlet B in spite of farmlet A supporting a 48% higher stocking rate by later in the trial period (2005). The study provided strong support for the proposition that intensive rotational grazing (farmlet C) provides more effective control of gastrointestinal nematodes than typical management (farmlet B) as evidenced by significantly lower WEC counts and anthelmintic treatment frequency. Tactical worm control based on routine monitoring of WEC provided adequate control of worms on all three farmlets for much of the experimental period but failed to prevent significant spikes in WEC to values associated with significant production loss on multiple occasions, and significant ewe mortality on farmlets A and B on one occasion.

The plasma concentration profiles of fenbendazole (FBZ), FBZ-sulphoxide and FBZ-sulphone were measured following intraruminal administration of FBZ at 7.5 mg kg−1 body weight in cattle and buffalo offered 3 different diets: 100% dry mature sorghum hay, 100% green Pennisetum spp. and a 50:50 mix of these 2 diets. Changing the diet from dry to green fodder resulted in significantly lower systemic availability of FBZ and its metabolites in both species. Buffalo had a lower systemic availability of the drug than cattle on the dry diet and the difference between species increased when the diet included green fodder. It is suggested that decreased transit time of digesta on the green fodder reduced systemic concentrations by reducing the time available for gastrointestinal absorption of the drug.

Swamp buffalo (Bubalus bubalis) and Droughtmaster cattle (Bos indicus x B. taurus), fitted with gastrointestinal cannulae, were dosed intraruminally with fenbendazole at 7.5 mg/kg liveweight, together with a chromium oxide capsule and a pulse dose of NaCoEDTA, to estimate the flow dynamics of the digesta in the rumen and duodenum. The concentrations of fenbendazole (FBZ) metabolites were measured in plasma and duodenal fluid collected over 120 h. In plasma, significantly lower peak concentrations and earlier disappearance of FBZ and its sulphoxide (OFZ) metabolite were observed in buffalo, which considerably reduced systemic availability in comparison with cattle. The availability of OFZ in the duodenal fluid of buffalo was significantly lower, whereas FBZ disposition was similar to that in cattle. The turnover rate of fluid in the rumen was higher in buffalo than in cattle, while the flow parameters for other digesta were similar in the two species. It is concluded that the decreased absorption of drug in buffalo was attributable to the shorter residence time of the dose in the rumen, and probably in the entire gastrointestinal tract. This may reduce the efficacy of treatment and indicate the need for higher dose rates for benzimidazole anthelmintics in buffalo than in cattle.

Small volumes of oxygen introduced into vials of medium at pH 7 prepared under anaerobic conditions reacted with the medium during sterilization by heating, and also raised the redox potential. The partial pressure of oxygen (pO2) in the medium, and the redox potential (Eh) were measured and their effect on the number of spores of Clostridium botulinum type E required to produce growth, and hence on the probability of growth from single spore inocula, was determined. In the presence of a pO2 of up to 4·5 × 10−3 atm resulting in an Eh of c. + 217 mV, the probability of growth from single spores within 5 days at 20°C was equal to that in strictly anaerobic conditions at an Eh of − 400 mV. At pO2 values of between 5·3 × 10−3 atm and 8·4 × 10−3 atm corresponding to Eh values of between + 226 mV and + 254 mV an inoculum of between 10 and 1000 spores was required to produce growth and at pO2 values of between 1·12 × 10−2 atm,and 1·6 × 10−2 atm, corresponding to redox potentials of between + 271 mV and + 294 mV, the number of spores required to produce growth was between 2 × 104 and > 2 × 105. The relationship between pO2 and Eh depends on the chemical nature of a culture medium or food, and in order to assess the probable influence of these parameters on growth of C. botulinum in a medium or a food it is necessary to determine both the redox potential and the partial pressure of oxygen.