Self-cure and immunity following infection and reinfection in ovine haemonchosis

Egg expulsion and post mortem examination were used to compare self-cure and resistance to reinfection with Haemonchus contortus in female Merino sheep, AA and BB hemoglobin types. Animals were infected by 500 or 5000 larvae of H. concortus once, or several times, before being challenged by 500 (low dose) or 5000 (high dose) larvae.In AA sheep infected once by 500 larvae self-cure was detected by a challenge of either 500 or 5000 larvae while in multi-infected sheep only a challenge of 5000 L₃ induced the self-cure. In BB sheep a mild self-cure was detected only by a challenge of 5000 larvae.In AA sheep receiving several infections, challenge by 500 larvae was not effective in producing resistance while 5000 L₃ was effective in producing a state of resistance for up to 35 days post challenge.At post mortem examination, a strong state of resistance was evidenced in multi-infected animals by the low number of L₄ and adults present in the abomasum up to the 35th day after challenge.In BB sheep no state of resistance was demonstrated by challenge with either 500 or 5000 L₃, even in multi-infected animals.     

The epidemiology and pathogenic significance of haemonchosis in a merino flock in East Africa

The epidemiology of H. contortus was studied for 21 months in 80 set-stocked Merino ewes and their grazing in an endemic area in Kenya. Observations included faecal egg counts (weekly), worm burdens (monthly), haematological indices and clinical signs. The levels of infective larvae on the pasture were estimated using tracer sheep.The classical acute form of haemonchosis was seen with equal intensity in both ewes and lambs during periods of high rainfall and self-cure was confirmed as a flock phenomenon which occured simultaneously on one occasion in sheep of all ages.Not previously recognised were the uniformity of the faecal egg counts in both ewes and lambs over long periods when the worm burdens fluctuated greatly, the marked decrease in worm burdens without the occurence of classical self-cure during periods when the intake of infective larvae was low, and the inverse relationship between the individual worm-weight and the total worm population. It was also concluded that one of the most significant features of haemonchosis is not the acute syndrome so characteristically associated with the disease, but the fact that a moderate infection of a few hundred worms persisting over a period of several months will produce chronic anaemia and, ultimately, severe loss of bodily condition and deathes in ewes and lambs grazing on poor quality pasture.     

Study on the pathogenicity of ostertagiosis due to Ostertagia circumcincta in sheep,in Iraq

The response of Awassi sheep to Ostertagia circumcincta, the most prevalent ovine strongylid species in Iraq, was studied.A dose of 100000 larvae of O. circumcincta (Iraqi strain) induced moderate clinical symptoms of ostertagiosis. These symptoms were correlated with elevated pH of the abomasal fluid and increased plasma pepsinogen levels. There was no evidence of larval inhibition since the majority of the fourth stage larvae (L₄) continued their development. No appreciable loss of worm population was observed and most of the parasites survived and exhibited a prolific egg-laying potential six moths after initial infection. Assessments of the pathophysiological changes were performed and correlated with parasitological and clinical observations.The Awassi breed of sheep, the most prevalent in the Arab Middle East, seems to be more susceptible to ostertagiosis than other breeds and there was no spontaneous self-cure in this breed.     

Epidemiology of gastrointestinal parasites of Awassi sheep in Iraq

Studies on the epidemiology of gastrointestinal nematodes of sheep, conducted at two different farms in the Baghdad area, in 1979, revealed two peaks in worm egg counts in ewes, one in spring and the other in autumn; the latter coincided with the lambing season. Similarly in lambs, two peaks of infection were apparent, but the spring peak was not so well defined. During the hot and dry summer months of Iraq, helminthic infection was at a low level. Faecal culture for larvae and their identification indicated the predominance of Haemonchus contortus infection in the ewes and that of Ostertagia and Trichostrongylus spp. in the lambs examined, Trichostrongylus infection predominating during the hot, dry period of the year. The self-cure phenomenon as well as preparturient rise in egg counts, as reported in other countries, was also observed during this study.     

The immune response of sheep to larval challenge with Ostertagia circumcincta and O. ostertagi.

One of three groups of sheep was challenged twice-weekly with infective-stage larvae (L3) of the sheep parasite O. circumcincta, another with the cattle parasite O. ostertagi while the third received no larval challenge. Positive faecal egg counts (FEC) and a rise in plasma pepsinogen levels were observed only in those animals given O. circumcincta. Anti-O. circumcincta L3 IgG titres were rapidly elevated during parasite challenge with either O. circumcincta or O. ostertagi. Throughout the experiment, no rise in anti-adult IgG titres or eosinophil numbers was observed in peripheral blood in any group. On evidence of self-cure of the trickle-infection, determined by a reduction in FEC, all groups were drenched and challenged with 15,000 O. circumcincta L3. No effect of previous challenge on parasite establishment or FEC was observed, although egg viability was significantly reduced in both groups given prior challenge. Significant differences in adult female worm length were observed between groups. Those recovered from animals previously challenged with O. circumcincta were shorter than from those given O. ostertagi which were in turn shorter than those from previously unchallenged animals. In utero egg counts were significantly lower in worms from animals previously challenged with O. circumcincta than in those from unchallenged control animals. The results indicate that a level of immunity to O. circumcincta can be conferred by exposure to O. ostertagi.     

Serum and abomasal antibody response of sheep to infections with Haemonchus contortus

Serum and abomasal IgA, IgG and IgM antibody response against adult worm, L3 and egg antigens of Haemonchus contortus was monitored by the ELISA technique after one or two infections with this nematode. Following the first infection, antibody levels in serum did not change materially. After administration of a challenge dose of infective larvae, antibodies of the three immunoglobulin classes in infected animals rose slightly, but this rise appeared later than the fall in the faecal egg counts. In contrast, in abomasal mucosa, IgA anti-larval antibody levels, which did not increase materially after the primary infection, rose rapidly after a transient inhibition when sheep were challenged. A close temporal relationship was observed between the rise in local anti-worm IgA antibodies and the self-cure reaction, but antibody levels fell rapidly after worm diminution. The local antibody response was thus considered to be related to immunity of sheep to H. contortus.     

Clinical evolution and radiographic findings of feline heartworm infection in asymptomatic cats

Clinical manifestations of heartworm disease in cats are variable; most cats seem to tolerate the infection well for extended periods. Heartworm-infected cats may undergo spontaneous self-cure due to the natural death of parasites without any symptomatology, or they may suddenly show dramatic and acute symptoms. Sudden death in apparently healthy cats is not a rare event. Thoracic radiographs are important tool for the diagnosis of cardiopulmonary disease. However, thoracic abnormalities are often absent or transient and highly variable in heartworm-infected cats. Findings, such as enlargement of the peripheral branches of the pulmonary arteries, with a varying degree of pulmonary parenchymal disease and hyperinflation, are the most typical features consistent with infection. A field study was performed for cats referred to the Veterinary Hospital Città di Pavia from January 1998 to December 2001 for routine health examinations and procedures to evaluate the clinical evolution and radiographic findings of feline heartworm infection. Thirty-four asymptomatic cats diagnosed with feline heartworm infection by antibody and antigen tests together with an echocardiogram that allowed worm visualization were included in the follow-up study. Cats were routinely examined every 3 months from the time of heartworm diagnosis until the outcome (self-cure or death). Self-cure was defined as no positive serology for heartworm antigens and no visualization of worms by echocardiography. A final examination for antibodies was carried after 12 months as a final confirmation of self-cure. Twenty-eight cats (82.4%) self-cured; including 21 that showed no clinical signs of infection throughout the study. Six cats died. The most common clinical features observed were acute respiratory symptoms and sudden death. Infection lasted over 3 years in the majority of the cats enrolled in the study. Thoracic radiograph appearance was variable, and the most commonly observed findings were focal and diffuse pulmonary parenchymal abnormalities.     

Application of western blotting procedure for the immunodiagnosis of visceral larva migrans in mice by using excretory/secretory antigens

A Western blotting procedure with excretory/secretory antigens from Toxocara canis larvae was developed for immunodiagnosis of visceral larva migrans in mice. In this study, eighty Swiss albino mice were allotted into two groups of 40 each as control and experimental groups, and T. canis ova containing infective larvae were given to mice in the latter group to form visceral larva migrans. Blood samples were taken from 5 infected and 5 control mice on days 25, 30, 35, 40, 45, 50, 55, and 60 after infection. After bleeding, the mice were necropsied. Slides were prepared from their brain tissues and examined for visceral larva migrans. Following this procedure, their guts were also examined for intestinal parasites. Protein bands of excretory/secretory antigens of 2nd stage larvae of Toxocara canis were determined by using SDS-PAGE. Sera from the mice were tested by Western blotting and results were compared to the protein bands obtained by SDS-PAGE to determine specific bands. Specific protein bands for visceral larva migrans were determined as 24, 28, and 48 kDa according to our test results.     

An enzyme-linked immunosorbent assay (ELISA) for detection of antibodies against Toxocara vitulorum in water buffaloes

Toxocara vitulorum, a parasite of the small intestine of cattle and water buffaloes, is mainly acquired by calves via the colostrum/milk from infected cows. To understand the development of immune responses in calves, antibody levels to a soluble extract antigen (Ex) from T. vitulorum infective larvae were measured by an indirect ELISA with sera of 15 buffalo calves, which were sampled every 15 days for the first 180 days after birth and 9 buffalo cows during the perinatal period. From all serum samples examined during the first 180 days, antibody level was lowest and highest in calves at 1 day of age before and after suckling colostrum, respectively, suggesting that the origin of antibodies was the colostrum. Immediately after birth, antibody levels in suckled calves remained at high levels until day 15, began to decrease to lower levels between 15 and 30 days and remained relatively stable until 120 days. By comparing the immune responses of these animals with their parasitological status it was considered possible to determine if passively acquired or actively produced antibodies provided protection against the infection. High numbers of T. vitulorum eggs in the feces between 30 and 60 days indicated that passively acquired antibodies did not provide protection against the infection, at least during these first days, and the maximum fecal egg counts during 30-45 days were coincident with decreased antibody levels. Between 60 and 120 days, when serum antibodies were detected at reduced, but stable levels, adult nematodes were expelled from the intestines and no more T. vitulorum eggs were found, suggesting development of acquired resistance. However, the potential and functional protective role of the antibodies against T. vitulorum infection and the process of self-cure requires further investigation.     

Effects of grazing undrenched weaner deer on chicory or perennial ryegrass/white clover pasture on the viability of gastrointestinal nematodes and lungworms

This study determined the in vitro effects on the viability of internal parasites of grazing undrenched weaner deer on either chicory (Cichorium intybus) or perennial ryegrass (Lolium perenne)/white clover (Trifolium repens) pasture. One experiment investigated the hatching and development of gastrointestinal nematode eggs and larvae, and the development and motility of L1 lungworm (Dictyocaulus eckerti) larvae, and a second experiment used larval migration inhibition assays to test the viability of L1 lungworm larvae extracted from the faeces of weaner deer grazed on either chicory or pasture when they were incubated with rumen and abomasal fluids from fistulated deer also grazing on chicory or pasture. The incubations were undertaken with and without added condensed tannins purified from chicory and with or without polyethylene glycol (PEG) to bind the tannins. Chicory had no effect on the hatching and development of gastrointestinal nematode eggs and larvae. Grazing chicory reduced the number of lungworm larvae developing to the L3 stage, and L1 lungworm larvae from the faeces of chicory-grazed deer were less viable in rumen and abomasal fluid than larvae from pasture-grazed animals. Abomasal fluid was significantly (P < 0.001) less inhibitory to the migration of L1 lungworms than rumen fluid. When the larvae were incubated in rumen and abomasal fluids from chicory-grazed deer, their passage through sieves was significantly (P < 0.001) reduced in comparison with when they were incubated in the fluids from pasture-grazed deer Adding condensed tannins to rumen fluid increased the inhibition of the migration of L1 lungworm larvae but PEG removed this inhibition; this effect was not observed with abomasal fluid.     

Growth Dynamics of White Muscle Fibres in Relation to Somatic Growth of Larvae of European Sea Bass (Dicentrarchus labrax, L.)

The body length and diameter of muscle fibres from the right hypaxial tail myomeres were measured in 100 sea bass larvae (Dicentrarchus labrax, L.) at ages of 40 to 90 days, from a fish farm in Croatia. The larvae were kept at a temperature of 19-20 degrees C and fed with standard feed for their respective ages. Body lengths of larvae 40-90 days old varied from 8 to 42 mm, increasing significantly in 10-day intervals (p < 0.001), except between days 50 and 60 when there was no increase in the body length. The measured diameters of muscle fibres ranged from 10 to 80 microm, the mean value increasing except between days 50 and 60. The fibre diameter in larvae aged up to 60 days was 10-40 microm, while that in larvae older than 70 days was 51-80 microm. In 90-day-old larvae there were no fibres of 10-20 microm in diameter; however, the number of fibers of 61-80 microm diameter increased. A statistically significant concordance was determined between the body length and fibre diameter of larvae 70 to 90 days old. Between days 50 and 60, arrested growth of the larval body and corresponding decrease in fibre diameter was observed. From day 45 until days 59, larvae were fed with Artemia nauplii and microparticles of formulated diets. From the day 60, minced fish was added to formulated diets. This change in diet probably resulted in the observed growth arrest.     

Efficacy of milbemycin oxime against fourth-stage larvae and adults of Ancylostoma tubaeforme in experimentally infected cats

The efficacy of milbemycin oxime against fourth-stage (L4) larvae and adults of Ancylostoma tubaeforme was investigated in a trial involving 24 young domestic shorthair cats. The animals were inoculated with approximately 300 infective stage three (L3) larvae and divided into three groups. After 12 days, eight cats (group 1) were treated with medicated tablets containing 4 mg milbemycin and 10 mg praziquantel to test the efficacy against L4 larvae; eight cats in group 2 were treated with the same tablets after 33 days to test the efficacy against adult worms; and eight cats in group 3 were treated with a placebo tablet. Faecal egg counts were determined periodically in each cat and after 40 or 41 days the number of worms in each animal was determined postmortem. The egg count reduction was determined by comparing the geometric mean numbers of eggs per gram of faeces in the placebo and medicated groups, and the worm reduction by comparing the geometric mean numbers of worms. The egg count reduction was more than 99 per cent in both treated groups, while the number of worms in groups 1 and 2 were reduced by 94.7 per cent and 99.2 per cent, respectively.     

THE EFFECT OF LICE INFESTATION ON THE GROWTH OF HEREFORD CALVES

SUMMARY Two experiments were carried out to examine the effects of cattle lice on the productivity of young calves. In both experiments the main species present was Linognathus vituli. In the first experiment, treatment of grazing heifers and their suckled calves did not improve the overall bodyweight gain of either the heifers or the calves. The calves experienced a rapid build-up in lice populations during autumn and early winter which was associated with mild transient pathogenic effects, but this was followed by an apparent ‘self-cure’ reaction and compensatory gain during late winter. In the second experiment young calves were placed in feedlots and fed high and low planes of nutrition. Treatment to remove lice did not result in any improvements in growth rate on either plane of nutrition; however it was clear that untreated, undernourished calves had heavier lice infestation.     

The influence of breed on the susceptibility of sheep of Haemonchus contortus infection in Kenya.

The relative resistance of six breeds of sheep to Haemonchus contortus infection was investigated under field conditions on two stations situated in a semi-arid zone in Kenya. As judged by faecal egg counts, monitored throughout a two-year period, it was found that there were consistent differences between the breeds. The breeds studied in order of increasing susceptibility were Red Masai, Blackhead Persian, Merino, Dorper, Corriedale and Hampshire. Post mortem findings confirmed that the differences in faecal egg count were related to the establishment of worms rather than larval inhibition or the suppression of worm egg production. There was also evidence that the Red Masai showed self-cure more frequently than the other breeds.     

Changing reactivity of caprine and ovine mononuclear phagocytes throughout part of the life cycle of Oestrus ovis: assessment through spontaneous and inductible NO production.

The in vitro reactivity of monocyte-derived macrophages (MDM) from Oestrus ovis (O. ovis) artificially infested lambs and kids was determined by measuring their production of nitric oxide (NO) during the course of infestation. In both species, crude antigenic preparations obtained from O. ovis first instar larvae (L1) were found to significantly (P < 0.01) inhibit this NO production, whereas O. ovis second instar (L2) extract stimulated it. Furthermore, this NO production by MDM decreased during infestation and was related to blood eosinophilia. It appears that crude antigenic extract from O. ovis modified the NO activity of macrophages from lambs and kids infested with O. ovis larvae.     

Experimental Trichinella spiralis infection in sheep.

During susceptibility studies of non-specific hosts, three merino sheep were infected with 3000, 5000 or 7000 Trichinella spiralis larvae by gavage. Clinical, physiological and serological parameters were assessed during the experiment. On the 152nd day p.i., animals were necropsied and, using artificial digestion methods, numbers of Trichinella larvae in muscle tissues were determined. The most infected parts were masseters with 3122 larvae g-1 muscle, 5526 larvae g-1 muscle and 4058 larvae g-1 muscle and diaphragms with 2778 larvae g-1 muscle, 2725 larvae g-1 muscle and 2320 larvae g-1 muscle, for the 3000, 5000 and 7000 infection levels, respectively. A positive correlation between infective rate and circulating antibodies was observed using ELISA and latex agglutination (LA) test methods. Trichinella larvae from sheep applied by gavage to ICR mice developed to the muscle stage. No significant changes were found in the clinical and physiological parameters of infected animals. Our results confirm the high susceptibility of merino sheep to T. spiralis infection.     

Effects of legume forages on ovine gastrointestinal parasite development, migration and survival

Lambs grazing certain legumes have reduced parasite intensities compared to lambs grazing ryegrass swards. Eighteen replicates of white clover (cv. AberHerald), lucerne (cv. Luzelle), red clover (cv. Merviot) and perennial ryegrass (cv. Abersilo) were sown at equivalent field rates in 25 cm diameter PVC pots and maintained outside for 6 months. On day 0, forage in each pot was cut to 50 mm from soil level and the pots were placed in a glasshouse (at 19-25 degrees C and 70% humidity) in a randomised block design. Ten grams sheep faeces containing 2,133 Haemonchus contortus eggs per gram were placed on the soil in each pot. Six replicates of each forage were destructively sampled on days 14, 21 and 29. Forage samples were cut at 50 mm from the soil surface and at the soil surface to give two samples per pot. The number of nematodes was determined by a modification of the Whitehead tray method. The ratio of free-living to infective-stage larvae was determined from at least 10% of the larvae. The number of H. contortus larvae kgdrymatter(-1) (DM) forage was calculated and the data rank transformed prior to analysis by ANOVA. There were fewer larvae on legumes compared with ryegrass on samples from forage above 50 mm (P<0.001) but there was no forage effect on larvae below this height. The sum of larvae present on all forage per kilogram DM showed fewer larvae on red clover compared with ryegrass on day 21 (P<0.05). There was an effect of day on the total number of larvae on forage (P<0.001) but there were no foragexday interactions. Analysis of the data according to the leaf area above 50 mm from the soil surface confirmed these results, that there were fewer larvae on legume forages than ryegrass above this height (P<0.01). Overall, red clover affected the development of H. contortus and all legumes affected larval migration above 50 mm compared with ryegrass but survival of larvae was similar on all forages. Further work is needed to determine if these effects of legume forages would reduce the number of parasitic larvae ingested by livestock under field conditions.     

Arthropod parasites of Hartmann's mountain zebra, Equus zebra hartmannae, in South West Africa/Namibia

Twelve Hartmann's mountain zebra, Equus zebra hartmannae, were shot for arthropod parasite recovery during the period June 1980-June 1981 on a farm in the Khomas Hochland region of South West Africa/Namibia. Four species of Gasterophilus larvae, 1 species of Rhinoestrus larvae and 3 ixodid tick species were recovered. The seasonal prevalence of the Gasterophilus species larvae and of Rhipicephalus evertsi mimeticus was determined. Three horses examined on the same farm were infested with larvae of 2 Gasterophilus species and with the same ixodid tick species as the zebras.     

Self-cure in zebu calves experimentally infected with Trypanosoma vivax

The course of an experimental infection of Zebu calves (6-12 months old) with Trypanosoma vivax was studied. Three of eight (38%) infected calves died, two within the first three weeks of infection and the other at 17 weeks. The remaining animals were self-cured. It would appear that self-cure of the infection was related to the ability of the calves to control parasitaemia as well as the severity of anaemia.     

Evaluation of Duddingtonia flagrans in reducing infective larvae of Haemonchus contortus in feces of sheep.

Consequences of nematode infections due to Haemonchus contortus are a serious constraint for the sheep industry worldwide. Development of anthelmintic resistance and increasing concern about the impact of anthelmintic use dictate the need of alternative control. Such an alternative is using the nematode trapping fungus Duddingtonia flagrans to reduce infective larvae levels on pasture. Two trials were conducted to determine the effect of D. flagrans in reducing infective larvae (predominantly H. contortus) in feces. The first trial determined the dose effect of D. flagrans in reducing infective larvae in feces. Eighteen ewes were dewormed to remove existing infections and randomly assigned to six treatment groups: 5 x 10(4), 1 x 10(5), 2.5 x 10(5), 5 x 10(5), 1 x 10(6) or no (control) spores of D. flagrans per kg of body weight mixed in their feed for 7 days. Fecal samples were collected daily from these and from infected donor ewes. Feces from individual-treated ewes were mixed with equal amounts of donor ewe feces, theoretically approximating oral dose spore concentrations of 2.5 x 10(4), 5 x 10(4), 1.25 x 10(5), 2.5 x 10(5), 5 x 10(5) and no spores, and were cultured. Across dosages and during the 7 days of fungus feeding, percent reduction of infective larvae ranged from 76.6 to 100.0%. The second trial determined the effect of D. flagrans at the dose of 10(5) spores per kg body weight on reducing infective larvae in feces from naturally infected lambs. Twenty lambs were randomly assigned to either treatment or control groups based on fecal egg count. Treatment lambs were fed spores mixed in feed for 7 days. Feces were collected daily and cultured. During the 7 days of fungus feeding, the percent reduction of infective larvae ranged from 82.8 to 99.7%. Results of these trials demonstrated that the nematode trapping fungus D. flagrans was highly effective in reducing infective larvae in sheep feces and should be considered as a biological control agent for integrated nematode control programs.