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The first structure-activity relationships of the anthelmintic cyclooctadepsipeptide PF1022A have been established via a systematic exchange of the leucine residues by a series of related N-alkylated amino acids. The data presented strongly suggest that (L)-N-methyl-leucine is crucial for high in vivo activity.
In two trials, lambs were trickle infected with 400 or 1000 infective larvae of Haemonchus contortus (HcL(3)), twice weekly for 4 weeks from the day of birth. Following anthelminthic treatment at 6 weeks (Trial 1) or 7 weeks of age (Trial 2), lambs were challenged 1 week later with a trickle infection totalling 5000 (Trial 1) or 10000 HcL(3) (Trial 2). In both cases, significant protection (P<0.05) ranging from 42 to 79% was achieved against egg and worm counts. Serum antibody responses as well as abomasal lymph node cell proliferation and production of interferon-gamma or interleukin (IL)-5 did not differ significantly between immunised and control lambs. The results are consistent with earlier findings that neonatal lambs can generate protective immunity against Trichostrongylus colubriformis, but the underlying mechanism(s) remain to be determined.
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The gerbil Meriones unguiculatus, infected with three species of nematodes, each located in a separate part of the gastrointestinal tract, provided a reliable laboratory assay for the evaluation of broad-spectrum anthelmintic activity. Gerbils harbouring 6-day-old infections of Haemonchus contortus, Trichostrongylus colubriformis and T. sigmodontis were given selected broad-spectrum anthelmintics by gavage. Three benzimidazoles, thiabendazole, oxfendazole and albendazole, a tetrahydropyrimidine, morantel, an imidazothiazole, levamisole hydrochloride, a macrocyclic lactone, ivermectin and an experimental natural product, paraherquamide, were active against all three nematodes at various dosages. Trichostrongylus colubriformis was most sensitive to levamisole hydrochloride, morantel, thiabendazole and paraherquamide whereas ivermectin, oxfendazole and albendazole were more effective against H. contortus. All compounds were active against the caecal nematode T. sigmodontis although it was less sensitive than T. colubriformis. Haemonchus contortus was more sensitive than T. sigmodontis to all anthelmintics tested except thiabendazole.
The incidence of haemonchosis in sheep and goats in Sierra Leone showed a seasonal variation with a high peak in the dry season (October to January) and a low one from March to May. Mean relative densities were significantly higher in young hosts and showed two peaks, a high one from August to December and a low one from April to June while the mean relative densities of old hosts were low and exhibited an irregular seasonal pattern with no defined peaks. The peak seasons in young hosts coincided approximately with the dry and rainy seasons. Male hosts showed an overall higher but not a significant mean relative density than females but for most of the months mean relative densities of infection of female hosts were not significantly higher than those of males. The roles of climatic conditions and arrested development (hypobiosis) and self-cure and host age immunity on Haemonchus contortus worm burdens are discussed.
Laboratory experiments were performed to investigate the ability of isolates from the predacious fungus species Arthrobotrys conoides (one isolate) and Al. robusta (four isolates) to trap and kill infective Haemonchus placei larvae. Three groups for each isolate were formed: group 1, fungi and infective larvae; group 2, fungi; group 3, infective larvae. There were statistically significant differences (P < 0.05) between the antagonistic effects of three isolates of A. robusta, but there was no such difference between the isolate of A. conoides and isolates of A. robusta. This result indicates that there can be a variation in the predatory capacity of different isolates within a single fungus species. After 20 days of assay, there was greater recuperation of infective H. placei larvae in group 3 compared to group 1.
The breed differences in post-parturient rise (PPR) in faecal egg counts in lambing ewes of different breeds naturally infected with Haemonchus contortus were compared. The ewes of Nali, 50% Nali x 50% Russian Merino/Corriedale and 37.5% Nali x 62.5% Russian Merino/Corriedale were treated with fenbendazole while ewes of the above three breeds as well as 25% Nali x 75% Russian Merino/Corriedale, were kept untreated. Observations from 3 weeks before lambing to 12 weeks post-lambing at weekly intervals revealed that in treated ewes, egg counts of Nali did not differ significantly with 50% Nali whereas 37.5% Nali had significantly higher egg counts than those of Nali and 50% Nali. In untreated ewes egg counts of 50% Nali were significantly higher than that of Nali on some occasions, while 25% Nali had significantly higher egg counts than the other three breeds. None of the ewes from the Nali breed showed signs of haemonchosis, whereas few lambing ewes of the three Nali crosses (50%, 37.5% and 25% Nali) showed signs of haemonchosis and one ewe of the latter breed died.
The influence of dietary protein supplementation upon resistance to haemonchosis was examined in Dorsimal (Polled Dorset x Malin) lambs offered two levels of protein. Lambs were offered either a complete basal ruminant diet (15% crude protein (CP)) or the same diet supplemented with fish meal as a source of rumen bypass protein (19% CP). Lambs from each dietary treatment group were given either a 7-week trickle infection with Haemonchus contortus infective larvae (L3) or remained uninfected. All lambs were drenched with anthelmintic at week 8 post-infection (PI), challenged with a single dose of 5000 H. contortus L3 one week later, and killed 14 days post-challenge (PC). Lambs on the supplemented diet that were trickle infected showed a significant reduction in egg output. Supplementation and previous infection did not affect either growth rate, worm burden, worm development or haematological parameters. There was a trend for enhanced growth among supplemented non-infected lambs in comparison to lambs which received the basal ration.
A single oral dose of an aqueous suspension containing 11,350,000 chlamydospores of a Mexican isolate of Duddingtonia flagrans (FTHO-8) given to sheep, resulted in a maximum reduction of 88% (range 86.7-90.4%) of the population of Haemonchus contortus infective larvae in the faeces. The effect of this treatment continued for 4-5 days after administration of the suspension. The possible use of this treatment as a method of control of ovine haemonchosis is discussed.
Five adult ewes and five lambs were repeatedly immunized with weekly doses of 10,000 irradiated Haemonchus contortus L3 before challenge with 100,000 and 10,000 normal larvae respectively. Two groups of non-immunized ewes and lambs were similarly challenged and one animal from each of the four groups killed on days 3, 5, 7, 10 and 24 post-challenge. The cellular changes in the abomasal mucosa were less marked in the non-immunized groups than in the immunized animals and appeared later in the lambs than in the ewes. Thus, in the immunized ewes increases in the numbers of mast cells and eosinophils were evident within five days of challenge whereas similar changes appeared later in the immunized lambs. Also marked lymphoid aggregates at the base of the mucosa and in the submucosa were detected only in the immunized ewes. However, both immunized and non-immunized ewes showed rises in the numbers of IgA plasma cells after challenge which were not evident in either group of lambs.
Haemonchus contortus originally derived from an Ontario field strain was used to infect laboratory rabbits. Both enshealthed and artificially exsheathed larvae were given at doses of up to 10 million larvae per rabbit. Only artifically exsheathed larvae were infective and a dose of at least 50,000 was required to produce a significant level of infection. H. contortus completed its development in the rabbit, but at a slower rate than in the sheep. Although a few fecund females were found 35 days after infection no eggs were observed on faecal examination. A high proportion of early fourth stage larvae were recovered from numerous rabbits including some which were examined well beyond a postinfection period when all worms should have been in the adult stage. This phenomenon occurred when freshly harvested and exsheathed larvae were used. Post-exsheathed cold conditioning of larvae at 5degreesC further increased the proportion of early fourth stage larvae (maximum proportions were found after 21 and again after 79 days of cold conditioning and thereafter declined). Whether such larvae were inhibited cannot be stated definitely. However, several of these larvae contained crystalline rods within their soma, and the structure of the crystals was similar to those previously described from inhibited H. contortus larvae recovered from sheep. It was concluded that the laboratory rabbit could be a useful host for further studies on some aspects of the biology and pathogenicity of H. contortus.
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Haemonchus longistipes specimens were collected from experimentally infected Sudanese goats and camels for morphological studies. The specimens from goats were much smaller than those from camels, but their infectivity to camels was not affected by their passage in goats. Goats inhabiting the same areas as camels in the Sudan were found to be naturally infected with H. longistipes, but sheep were not and were resistant to experimental infection.
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Adult Haemonchus contortus worms simultaneously excrete and fix CO2. Their initial content of CO2 was measured as 4.55 mumoles/100 mg wet weight and their excretion rate in air as 1 mumol/100 mg wet weight/h for at least 4 h. When the worms were incubated either aerobically or anaerobically with 14CO2 most of the metabolized radioactivity was associated with propan-1-ol and propionate but small amounts were found in succinate and lactate. No radioactivity was associated with ethanol or acetate, two major catabolites of glucose. Stepwise degradation of the metabolized radioactive propanol and propionate showed that all the radioactivity in both compounds was associated with carbon atom no. 1. These results show that H. contortus has much in common with the anaerobic energy metabolism of Ascaris lumbricoides but they are not inconsistent with the utilization of the tricarboxylic acid cycle by the worm. H. contortus worms were found to metabolize their excretory products. When they were incubated with either [2,3-14C]succinate or [2-14C]acetate, 14CO2 was excreted under aerobic but not under anaerobic conditions. These results are consistent with a pathway similar to that used by Ascaris operating aloneunder anaerobic conditions and together with the tricarboxylic acid cycle under aerobic conditions.
The response of Awassi sheep to primary infection with 10 000 Haemonchus contortus larvae and subsequent challenge with 20 000 larvae was studied. Two types of response to re-infection were observed. The first, characterized by a sharp decline in egg counts, progressive improvement in packed cell volume (PCV) and a marked loss of adult worms and challenge larvae, was considered as self-cure and protection. In the second type of response there was a temporary decline in egg counts, no improvement in PCV and necropsy findings indicated hyper-infection with challenge larvae.
Abomasal and duodenal motility was investigated by electromyography in 4 lambs fitted with permanent abomasal and duodenal cannulae and infected twice with 25,000 Haemonchus contortus larvae at an interval of 39 days. In addition, abomasal and duodenal pH, flow rate of digesta of the duodenum, and faecal water content were determined. The 80-90 min cycles of antral contractions were shortened for at least 3 weeks after infection. This was accompanied by a 26% increase in the number of duodenal migrating myoelectric complexes/day during the intraparietal development of the larvae (10-12 days). Concomitantly, the duodenal flow of digesta increased from 215 +/- 22 ml/h to 318 +/- 15 ml/h 4 days after infection and was associated with a higher water content of the faeces. The frequency of the migrating myoelectric complexes, but not the duration of antral motor cycles (or the duodenal flow rate), was correlated with both gastric (P less than 0.01) and duodenal (P less than 0.05) pH after the first and second infections. The duodenal flow rate was correlated with the Cl- concentrations of the abomasal contents (P less than 0.01) and with the duodenal pH (P less than 0.05). It is concluded that, (i) the duodenal motor disturbances associated with experimental H. contortus infection are related to the alterations in abomasal gastric acid secretion which occurs during the histotrophic phase of larval development and (ii) the increased duodenal flow is a consequence of ionic permeability changes of the gastro-intestinal mucosae.
Inhibited development of Haemonchus contortus was studied in single experimental infections of worm-free lambs. Chilling of the infective larvae at +4 degrees C was without effect on the percentage of larvae subsequently becoming inhibited and a period of exposure to autumnal conditions was unnecessary to induce a high rate of inhibition. It was concluded that seasonal inhibition of H. contortus in East Anglia is brought about primarily by an environmental stimulus acting upon the preparasitic stages but that, unlike Obeliscoides cuniculi and Ostertagia ostertagi, this was not cold. It could be provided in a culture kept in the dark at 25 degrees C for 12 days. While the age of the host did influence the phenomenon, in that larvae were less inclined to inhibition in very young animals, it was concluded that this was not a primary factor in the aetiology.