Parasite antigens and host antibodies in Ostertagia circumcincta infection of the sheep

and 1972. Parasite antigens and host antibodies in Ostertagia circumcincta infection of the sheep. International Journal for Parasitology, 2: 449–457. An allergenic component was separated from Ostertagia circumcincta antigens and specific reaginic antibody was separated from the corresponding 7S antibodies in sheep sera. Further evidence was obtained that the immunoglobulin class defined as IgG1A contains the reaginic or homocytotropic antibodies in sheep. Both the IgG1A antibody and P.C.A. levels continued to increase after the expulsion of the parasites, whereas the levels of anti-Ostertagia IgG1 did not.     

The statistical distribution of trichostrongylid nematodes in grazing lambs

Barger I. A. 1985. The statistical distribution of trichostrongylid nematodes in grazing lambs. International Journal for Parasitology15: 645–649. Worm counts of the genera Haemonchus, Ostertagia, Trichostrongylus and Nematodirus from 104 naturally-infected lambs were found to follow negative binomial distributions. Although mean counts covered a 10-fold range, k values did not differ significantly among the four genera. It was concluded that all genera were equally overdispersed. Some implications of this overdispersion for flock productivity and for parasite control were examined     

Cryopreservation of the first-stage larvae of trichostrongylid nematode parasites

First stage (L1) larvae of Haemonchus contortus, Trichostrongylus colubriformis and Ostertagia circumcincta can be cryopreserved in the presence of DMSO using a two-step freezing protocol involving an initial period at −80°C prior to transfer to liquid nitrogen. Thawed L1 larvae continue development in vitro producing third stage (L3) larvae that are infective to sheep when dosed per os. Establishment rates for L3 larvae grown from thawed L1 larvae were 40 and 80% for H. contortus and T. colubriformis, respectively. There was no difference in survival or infectivity between benzimidazole (BZ)-susceptible and BZ-resistant H. contortus isolates and cryopreservation caused no shift in their BZ-resistance status as indicated in an in vitro larval development assay. Cryopreservation also had no effect on the sensitivity of these isolates to the avermectins or levamisole in vitro. High survival rates (60–70%), good levels of establishment and the stability of anthelmintic resistance status of isolates indicate that little if any selection occurs during the cryopreservation process. L1 larvae of all 3 species have been successfully recovered after 16 months storage in liquid nitrogen, cultured to the L3 stage and established in sheep.     

Stage-specific cuticular proteins of Ostertagia circumcincta and Ostertagia ostertagi

In this study we have shown that NHS-biotin and I¹²⁵-streptavidin can detect cuticular polypeptides of Ostertagia spp. The labelled polypeptide profile of intact nematodes is simple compared to the profile obtained by labelling homogenates. None of the major internal polypeptides are labelled and the subset of proteins labelled in intact nematodes appears to be mainly surface associated. The results presented here demonstrate that NHS-biotin may be used as a reagent for the analysis of surface polypeptides. The surface polypeptide profiles of the five major developmental stages (L1, L2, L3, L4 and adult) of Ostertagia circumcincta show a series of stage-specific molecules with no polypeptides common to all stages, indicating that the cuticle is a dynamic structure which changes throughout the life cycle. Similarity comparison of Ostertagia ostertagi L3 and L4 stage surface profiles showed that each stage is clearly distinct; comparison of these stages between the two species shows an overall similarity.     

A comparison of the periparturient rise in fecal egg counts of exotic and domestic ewes

Ewes of three exotic breeds (Florida Native, Barbados Blackbelly and St. Croix) showed no periparturient rise in fecal egg counts (PPR) when housed from late fall through lambing and weaning. Domestic breed ewes (Rambouillet and Finn-Dorset × Rambouillet) showed a pronounced PPR 6–7 weeks post-lambing while St. Croix × domestic ewes showed an intermediate PPR under similar conditions. There was no PPR in non-lambing ewes.In a second experiment, breed differences in the PPR were not as pronounced when ewes were allowed to graze a contaminated pasture after lambing. Florida Native, St. Croix and $\text{3}\text{4}$ St. Croix × $\text{1}\text{4}$ domestic showed lower PPRs than the remaining breeds (Barbados Blackbelly, F₁ hybrids of exotic and domestic breeds and domestic ewes), but the magnitude of the PPR was generally small in all breeds. No relationship was observed between hemoglobin genotype and PPR in either experiment.It was concluded that a low or absent PPR is an important manifestation of breed resistance in exotic ewes and that this may be useful as a marker for breeding parasite-resistant sheep.     

Development of simultaneous resistance in Ostertagia circumcincta to thiabendazole, morantel tartrate and levamisole

Le Jambre L.F., Southcott W.H. and Dash K.M. 1978. Development of simultaneous resistance in Ostertagia circumcincta to thíabendazole, morentel tartrate and levamisole. International Journal for Parasitology8: 443–447. A field strain of Ostertagia circumcincta was divided into five strains based on anthelmintic selection in the laboratory. The first strain was selected with 50 mg/kg thiabendazole, the second with 4 mg/kg morantel tartrate, the third with 3.2 mg/kg levamisole, the fourth was not selected and the fifth strain was selected with all three anthelmintics in each generation. The present paper reports the dose response of the eighth generation of the multi-selected strain to thiabendazole, morantel tartrate and levamisole and compares these results with those from the eighth generation of the single selected and unselected strains.In adult O. circumcincta the LD₉₅ for thiabendazole, morantel tartrate and levamisole was 172.0, 9.2 and 8.4 mg/kg respectively in the multiselected strain, compared with corresponding values of > 200, 6.1 and 6.9 for the single selected strains and 14.5, 2.8 and < 1.6 in the unselected parent strain.Multiple selection with three anthelmintics was associated with an increase in O. trifurcata from less than 0.1 % in the unselected strain to 16% in the multi-selected strain.An increase in inhibition was a feature of both multiple selection and selection by levamisole alone. The parent strain had less than 0.1 % inhibition but the incidence increased to 16% in the levamisole selected strain and to 2% in the multi-selected strain. Approximately 8% of the inhibited larvae in the levamisole selected strain were resistant to dose levels of levamisole from 1.6 to 8.0 mg/kg. Resistance in inhibited larvae was further enhanced in the multi-selected strain and after eight generations 100% of larvae in this strain were resistant to all dose levels up to 100 mg/kg thia-bendazole, 20 mg/kg morantel tartrate and 8 mg/kg levamisole. Apparently when selected with anthelmintics that are less effective against larvae than adults, O. circumcincta responds by increasing the percentage and resistance of inhibited larvae.     

Effects on plasma pepsinogen, gastrin and pancreatic polypeptide of Ostertagia spp. transferred directly into the abomasum of sheep

Ostertagia spp. obtained at slaughter from the abomasa. of previously infected sheep and transferred via cannulae into the abomasa of previously worm-free sheep produced severe effects. This was shown with populations of mainly adult parasites given to three sheep and also when mixed populations of 4th stage larvae and adults were given to three other sheep. Although more severe effects were produced when both larval and adult worms were transferred, there were clearly effects when isolates composed predominantly of adults were transferred. Sheep of each group had reductions in food intake and elevations in plasma pepsinogen and gastrin within 24 h of transfer of the parasites. Further increase in plasma pepsinogen and gastrin occurred when abomasal pH rose 5–7 days after infection. Plasma pancreatic polypeptide varied but showed a general tendency to be markedly lower after infection. Ostertagia spp. eggs were detected in the faeces of infected sheep 48 h after transfer. The proportions of adults and inhibited larvae recovered from the abomasa were similar to those in the donor sheep, showing that inhibition, or arrested development of larvae, continued after their transfer to worm free sheep.     

Manifestations of resistance to Haemonchus contortus in sheep: Worm populations and abomasal changes in sheep superinfected with 1,000,000 larvae of H. contortus

The super-infecting dose produced a marked rise in gastric pH in all sheep from the 3rd day after administration of larvae. Expulsion of the existing population of adult worms may have begun on the 4th day but was still only completed in 3/6 sheep on the 5th day. The larvae caused extensive damage in the individual glands which they parasitised. Very few of the 10⁶ larvae survived for 27 days and only in 1/8 sheep had they developed beyond early 4th stage at 27 days. Extensive histological changes were seen in the fundic mucosa beginning as early as 2 days after the superinfection. While the pH change preceded expulsion of the adults and was consistent in its timing, the timing of the expulsion was irregular. This throws doubt on the hypothesis that the change in physico-chemical conditions produced by the superinfecting larvae is the only cause of the expulsion of the adult worms.     

Development of anthelmintic resistance in nematodes from sheep in Australia subjected to different treatment frequencies

Development of anthclmintic resistance in nematodes from sheep in Australia subjected to different treatment frequencies. International Journal for Parasitology13: 125–132. In a three year (1978, 1979, 1980) field experiment in south-eastern Australia Merino weaner sheep were exposed to one of four worm control programmes. Sheep were treated either 50–51 times, 9–12 times, 3–6 times or 1–3 times annually with anthelmintic. Thiabendazole (TBZ) was used throughout 1978 and for the initial dose in 1980, levamisole (LEV) throughout 1979 and 1980. Sheep were replaced annually each December.During 1978 in vitro egg hatch tests indicated that populations of Haemonchus contenus with resistance factors to TBZ of 10·7, 10·0, 2.5 and 1·8 respectively were selected by these treatment frequencies. No resistance to LEV was detected in 1979 but during 1980 in vivo egg count reduction tests showed that highly resistant populations of Ostertagia spp. were selected in sheep dosed either 49 or 11 times. This was confirmed by an in vitro larval paralysis test. Partial resistance occurred where sheep were dosed 5 times but not where one dose was given. The only population of H. conlortus still present at the end of 1980 in sufficient numbers to allow eggs to be tested in vitro was from the sheep dosed once. This had retained its resistance to TBZ.     

Population dynamics of the parasitic stages of Ostertagia spp. in sheep

Callinan A. P. L. and Arundel J. H. 1982. Population dynamics of the parasitic stages of Ostertagia spp. in sheep. International Journal for Parasitology12: 531–535. The development and survival of continuing infections of Ostertagia spp. in weaner sheep were studied in order to develop a general model of the parasitic stages of the life cycle of these sheep nematodes. After 10 days, 13.8% of infective larvae (L₃) given at the rate of 1 dose of 1000 L₃ twice per week (group 1) and 20.8% of L₃ given at the rate of 10,000 L₃ twice per week (group 2) were recovered in the first of the serial nematode counts. In the final counts at 137 days, 7.7 and 0.7% were recovered in these groups. The build up and maintenance of nematode populations was regulated and related to the level of infection. A model in which the death rate of the parasitic stages was a function of the time of exposure to infection and rate of infection was used to describe the serial total nematode counts. During the experiment there was no noticeable trend in numbers of fourth stage larvae (L₄) in nematode counts, the size of adult nematodes, nematode egg counts (EPG) and egg output per female nematode (EPF). After 112 days, liveweight gains were significantly reduced in group 2 only, but increases in wool lengths were significantly reduced in both groups.     

Nervous regulation of abomasum motor activity in sheep

Pharmacological studies of abomasum motor activity were carried out in 3 sheep with chronic fistulae in the rumen and the fundus and pyloric part of the abomasum. The contractions of these parts of the stomach were recorded by the balloon method. The obtained results showed that in the abomasum stimulation of alpha-adrenergic receptors raises the degree of motor activity while stimulation of beta-adrenergic receptors inhibits it. It has been shown also that the sympathetic system has a greater influence through the beta-adrenergic receptors on the motor activity of the pyloric part than on that of the fundus of the abomasum. The analysis of the records obtained after bilateral vagotomy demonstrated that the automatism of the pyloric part was greater than that of the fundus of the abomasum.     

Acetylcholinesterase secretion by parasitic nematodes. IV. Antibodies against the enzyme in Trichostrongylus colubriformis infected sheep

Sheep infected with the nematode parasite Trichostrongylus colubriformis showed anti-T. colubriformis acetylcholinesterase. (AChE) antibodies in the IgG₁ but not the IgG₂ or IgM fractions prepared from their serum. Using the fluorescent antibody technique with representative sera, antibodies in the IgG₁ fraction exhibited specificity for antigens in the subventral glands of the worm excretory system. IgA antibody specificity for antigens in the excretory glands and intestine of the worm was also demonstrated.     

Eight types of endocrine cells in the abomasum of sheep

In the ovine abomasum, 8 types of endocrine cells were classified at ultrastructural level. The gastric-type EC cells contained oval and pleomorphic granules with high electron density. The intestinal-type EC cells were filled with oval, irregular and highly dense granules. ECL cells contained irregular granules with high density and wide clear spaces. D cells were filled with round granules showing low to moderate density and finely granular matrix D1 cells contained round or oval granules with variable, low to moderate density and finely granular content. G cells showed round and oval granules with moderate density, densely packed or flocculent content. F cells were filled with oval or elliptic granules showing low density with a finely granular and flocculent matrix. X cells contained round granules with high density and homogeneous material. Gastric-type EC cells, intestinal-type EC cells, D cells, and D1 cells were represented in the cardiac, fundic and pyloric gland areas of the ovine abomasum. ECL cells and F cells were confined to the fundic glands, G cells and X cells to the pyloric glands.     

The relevance of in vitro anthelmintic screening tests employing the free-living stages of trichostrongylid nematodes

The response of the free-living stages of Nippostrongylus brasiliensis, Nematospiroides dubius, Haemonchus contortus, Trichostrongylus colubriformis and Ostertagia ostertagi to a wide variety of antiparasitic agents in vitro was investigated. All the major broad spectrum veterinary anthelmintics showed good activity against each of these worms with EC50 values varying from about 0.0002 mg/l for certain benzimidazoles and ivermectin to about 6.5 mg/l for febantel. Of 22 known narrow spectrum anthelmintics useful only against H. contortus and/or helminths other than trichostrongyles, only 10% showed good activity at concentrations equal to or less than 10.0 mg/l. Further, only one of 15 antiprotozoal agents showed good activity in these tests at the 10.0 mg/l level. The screening test employing free-living Nippostrongylus brasiliensis was selected for an extended trial where the evaluation of 1400 miscellaneous organic chemicals was undertaken. Approximately 10% of these showed activity at concentrations equal to or less than 10.0 mg/l. It is concluded that in vitro screening tests employing the free-living stages of these five genera of nematodes afford simple yet effective means for selecting relevant compounds for further evaluation as possible leads to new broad spectrum anthelmintics for use in ruminants. However, tests using the free-living stages of these worms, including H. contortus, are unsuitable for detecting narrow spectrum 'specifics', e.g., for the treatment of haemonchiasis.     

Measurement of changes in parietal tonus of the abomasum in sheep

An inflatable bag of 250 ml at the pressure of 2 mm Hg inside the abomasal lumen was used to identify the abomasal smooth muscle tone. Reduced tone of the ovine abomasum was assessed, like for the fundic part of the canine stomach, in relation with the cyclic motor activity of the proximal duodenum. Opposite effects were obtained following parasympathomimetic drug injection versus peptides of the gastrin group, suggesting a high responsiveness of the abomasal smooth muscle tone to both neural and hormonal influences.     

Epidemiological studies on horses infected with nematodes of the family Trichonematidae (Witenberg, 1925)

Observations are reported on seasonal changes in the age structure of populations of nematodes of the family Trichonematidae recovered at regular intervals from the lumens of the large intestines of horses slaughtered in S.W. England. The results show that changes in the size of parasitic populations of Trichonema nassatum follow seasonal variations in the rate of infection, more individuals maturing during summer/autumn than during winter/spring because of proportional differences in the numbers of infective larvae ingested from the pasture. In contrast, larvae of T. longibursatum, T. catinatum and T. goldi ingested by grazing horses during summer accumulate in the gut wall rather than promptly returning to the lumen and developing to maturity. Their development is apparently inhibited until the following spring when 4th-stage larvae emerge en masse and quickly reach the adult stage. The relationship between fluctuations in the size of the adult populations of the 4 species studied and the characteristic seasonal variations in horse faecal egg counts is discussed.     

Mitochondrial diversity and the origin of Iberian sheep

Mitochondrial DNA diversity was analysed in 19 Iberian and six foreign sheep breeds. Three mtDNA lineages (B, A and C) were found in the Iberian sheep, with type B clearly predominating over the others. The results were analysed for each of the morphologically determined breed groups in Iberian sheep: Merino, Entrefino, Churro and Iberian trunks. MtDNA lineage C was found only in the Iberian trunk composed of Montesina and Ojalada. These two populations had high mtDNA variability, and in the Iberian sheep only Merino Branco had more variation. The other three Merino types studied showed moderate variability, including the most authentic Merino, the Spanish Merino. These three Merinos clustered closely in a multidimensional scaling representation of distances, while the fourth breed (Merino Branco) showed a clear separation. As for the other two trunks, breeds from the Churro group showed greater maternal uniformity while results for populations included in the so-called Entrefino trunk seemed to have a more heterogeneous maternal origin. The results obtained are discussed with available data from nuclear markers and with morphological classifications, and all this information is analysed in relation to the origin of the different Iberian sheep breeds.