Pilobolus species and rapid translation of Dictyocaulus viviparus from cattle faeces

In a series of five laboratory experiments observations were made on the role of Pilobolus in the translation of infective lungworm larvae from faeces of cattle. The results indicated that a substantial proportion of the lungworm larvae present in the faeces may be translated from the faeces by this fungus within eight days at a temperature of 15 degrees C. No clear relationship was observed between the numbers of Pilobolus and the translation of lungworm larvae. Further a longitudinal study on the occurrence of Pilobolus on faecal pats of grazing calves showed that between the beginning of July and the middle of September peak emergence of sporangia generally occurred within one week and most sporangia emerged within three weeks. From faecal pats which had been deposited at the end of September and the middle of October emergence of sporangia was lower and mainly occurred after two to four weeks.     

Spread of equine lungworm (Dictyocaulus arnfieldi) larvae from faeces by Pilobolus fungi

Between 10 and 25% of the Dictyocaulus arnfieldi larvae excreted in faeces from a naturally infected donkey were harvested as infective stages from faecal cultures by means of Pilobolus fungi. The faeces were collected between 24 and 56 hours after drenching the donor animal with Pilobolus spores and kept at 16 +/- 2 degrees C. Most larvae were collected between the 5th and the 8th day of culturing during which period fructification and sporangium discharge also peaked. The sporangia and the adhering larvae were collected in Petri dishes inserted between the faecal mass and a light source. All recovered larvae were viable. A mean larval length of 368 microns (range 312-440 microns) and width of 14.6 microns (range 12-20 microns) was recorded for the infective stage. The method was found suitable for the recovery of infective stages for experimental purposes. The authors suggest that the Pilobolus mechanism play an important part in the spread of equine lungworm infection under field conditions similar to the situation in bovine lungworm (Dictyocaulus viviparus) infection.     

Direct measurement of dispersal of Dictyocaulus viviparus in sporangia of Pilobolus species

Dispersal of Dictyocaulus viviparus larvae by Pilobolus sporangia was studied on 29 faecal pats deposited between the end of June and late October 1988. Faecal pats were covered daily from day 3 to 4 after deposition with a large petri dish to measure the numbers of sporangia released and the numbers of larvae carried. The yield of both was variable. Dispersal of lungworm larvae was lowest on over-grazed pasture or when Pilobolus growth was very poor. When faecal pats were sheltered by a long sward, 17 per cent or more of larvae present at deposition were transported in this manner. In July and August, peak dispersal of lungworm larvae was on day 5, in September on day 6 and in October on day 7, the increasing time intervals being probably associated with decreasing temperature.     

Epidemiology of bovine dictyocaulosis in Denmark

A field experiment was conducted over two grazing seaons with calves on a permanent pasture in order to follow the pattern of infection with Dictyocaulus viviparus. Infective larvae persisted during the first, but not during the second, winter of observation. By means of the agar-bile herbage technique, a moderate first peak of infection was demonstrated in the pasture 2–3 weeks before the appearance of respiratory signs in the calves. Fluctuations in faecal larval output were reflected in the herbage contamination with infective larvae close to faecal pats. This, as well as the horizontal dispersion of larvae in the pasture, took place in less than a week. The proportion of lungworm larvae recovered away from faeces was low during a period of dry and hot weather while herbage sampling at two-hour intervals during two days showed an increase in herbage contamination with lungworm larvae, but not with trichostrongyle larvae between 10 a.m. and 12 noon.The infectivity of the pasture was monitored by tracer calves and compared with the results of the pasture sampling. The general course of the infection in the calves and in the pasture was the results of interaction between them. In addition, the pasture infection was influenced by climate and the infection in the calves by the development of immunity. The course of infection in individuals appeared to have an influence on the general course of the infection through the contamination of the pasture.     

Studies on immunisation of suckling calves with dictol

The immunisation of young milk-fed calves with two doses of the commercially available Dictyocaulus viviparus irradiated larval vaccine, Dictol, was studied. In the first experiment, vaccination of groups of pail-milk-fed calves at 3 and 7 weeks of age resulted in 96.7% reduction of lungworm burdens upon challenge when compared with non-vaccinated controls. Contemporaneous vaccination of calves aged 8 and 12 weeks resulted in a reduction in lungworm burden of 98.9% compared with controls of the same age. The clinical reaction upon challenge of both age groups was minimal compared with their respective controls.In the second experiment a group of six suckling calves were vaccinated with Dictol at 3 and 7 weeks of age and then challenged with infective larvae of D. viviparus together with non-vaccinated controls. At subsequent post-mortem examination the mean lungworm burdens of the vaccinates was 87% less than that of the controls. Some clinical reaction evidence by increased respiratory rates and bodyweight loss occurred in the vaccinates but this was not as severe as in the controls. At necropsy a concurrent pneumonia due to Mycoplasma spp. was present and it is suggested that this contributed to the poorer protection obtained compared with the pail-milk-fed calves of the fist experiment.In the third experiment a group of suckling calves were again vaccinated at 3 and 7 weeks of age and then exposed to a natural challenge together with parasite-naive controls. Although the level of challenge was relatively low, immunisation was apparently high successful as lungworms were completely absent in the vaccinates when examined at necropsy.In all three experiments numerous pulmonary lymphoid nodules, which are a useful guide to the immune status of calves, were present in the vaccinates and absent or scarce in the controls. It is concluded that the cumulative evidence from the three experiments suggest that immunisation against infections of D. viviparus of young milk-fed calves in the field may be a practical proposition.     

The therapeutic efficacy and prophylactic activity of doramectin against Dictyocaulus viviparus in cattle

SUMMARY Three groups of 8, 4-month-old male Jersey or Jersey-cross calves were infected with 2400 Dictyocaulus viviparus L₃ larvae and either left untreated or injected subcutaneously with 200 μg/kg doramectin 5 or 25 days after infection (DAI). Lungworms were found in all untreated cattle (geometric mean = 49) at necropsy 39 or 40 DAI. None was found in any of the treated cattle. In a second experiment, groups of 6, 8-month-old calves were untreated or injected with 200 μg/kg doramectin 28, 21 or 14 days before each calf was challenged with 2700 D viviparus larvae. Lungworms were recovered at necropsy 32 to 34 DAI. The geometric mean worm burden in the untreated cattle was 550. This was reduced by 100%, 99.5% and 94.1% in calves treated with doramectin 14, 21 or 28 days, respectively, before infection. It was concluded that doramectin is a highly effective anthelmintic against D viviparus adult or L₄ infections of cattle, and that reinfection of treated cattle will be significantly reduced for at least 28 days after treatment.     

The relative susceptibility of Soay and Blackface sheep to natural infection with Dictyocaulus filaria.

Simultaneous studies by faecal larval counts on a mixed flock of Blackface and Soay ewes, yearlings and lambs suggested that the Soay animals were less able to develop resistance to Dictyocaulus filaria. This susceptibility resulted in a greater proportion of the Soay ewes developing heavy burdens of these lung worms, and in a greater proportion of the yearlings and ewes excreting D filaria larvae in their faeces.     

Spread of infective Dictyocaulus viviparus larvae in pasture and to grazing cattle: Experimental evidence of the role of Pilobolus fungi

Four calves experimentally infected with Dictyocaulus viviparus were made Pilobolus-free by hygienic measures and by feeding them irradiation sterilized feed. Two of the calves were orally administered laboratory cultured Pilobolus sporangia daily. As a result, the faeces from one air contained D. viviparus larvae and Pilobolus spores, and the faeces from the other pair contained D. viviparus larvae, but no Pilobolus spores.

Two identical plots were used for deposition of the two kinds of faeces, and one of them remained free of Pilobolus fructification. Herbage sampling and the use of tracer calves revealed that on this plot the larval contamination and the infectivity of the pasture were greatly reduced. A mean larval count of 1321 near the faecal pats (0–5 cm) in the plot where Pilobolus was observed was reduced to 69 per kg of herbage on the Pilobolus-free plot. At a distance of 100 cm from the pats, a reduction from 99 to 3 larvae per kg herbage was found.

Each plot was grazed by four parasite-free tracer calves for 3 days. During the subsequent stabling period of these calves, the lungworm larval excretion of those from the Pilobolus-free plot was reduced by 90% and the clinical symptoms were milder than those which grazed the plot which contained the fungus. The mean post mortem worm counts after 4 weeks of stabling showed a reduction from 167 to 25 worms.

A more marked effect of Pilobolus fungi on the transmission of D. viviparus infection is to be expected under field conditions where calves are grazing more selectively than in the present study.     

Verminous Pneumonia in Adult Dairy Cows in Southern Ontario due to Dictyocaulus viviparus

An outbreak of verminous pneumonia due to Dictyocaulus viviparus in a herd of mature, lactating, dairy cows in southern Ontario is described. The fact that the herd had been closed to new additions for ten years and had never experienced clinical disease due to D. viviparus in the past makes the occurrence of this herd problem difficult to explain. Correlation of fecal Baermann analysis for D. viviparus larvae with the progress of anthelmintic treatment is discussed. It is suggested that certain climatological variations in combination with unique, immunological aspects of D. viviparus infection may have contributed to the development of clinical disease in this herd.     

Nematode control in suckler beef cattle over their first two grazing seasons using a targeted selective treatment approach

Background

With concerns over the development of anthelmintic resistance in cattle nematode populations, we must re-examine our approach to nematode control in cattle. Targeted selective treatments (TST), whereby individual animals are treated instead of entire groups, are being investigated as an alternative. The study objective was to determine if anthelmintic usage could be reduced using a TST-based approach to nematode control in spring-born suckler beef cattle over their first and second grazing seasons (SGS) without affecting performance. In the first grazing season (FGS), 99 calves with an initial mean (s.d.) calf age and live weight on day 0 (June 28^th 2012) of 107 (23.1) days and 160 (32.5) kg, respectively, were used. The study commenced on day 0 when calves were randomised and allocated to one of two treatments; 1), standard treatment (control) and 2), TST. Control calves were treated subcutaneously with ivermectin on days 0, 41 and 82 in the FGS. All calves were treated with ivermectin on day 124 and housed on day 133. In the SGS, only heifer calves from the FGS were used and control heifers were treated with ivermectin on day 393. Animals were weighed, blood and faecal sampled every three weeks. The TST animals were treated with ivermectin if thresholds based on a combination of plasma pepsinogen concentrations, faecal egg count and/or the presence of Dictyocaulus viviparus larvae in faeces (FGS only) were reached.

Results

No TST calves reached the treatment threshold criteria in the FGS. The FGS average daily live weight gain (ADG ± s.e.m.) for control and TST group calves was 0.89 ± 0.02 kg and 0.94 ± 0.02 kg day⁻¹, respectively (P = 0.17). In the SGS, all heifers were treated with ivermectin on day 431 due to clinical signs of respiratory disease. The ADG for control and TST heifers from turnout on day 321 to day 431 was 0.90 ± 0.04 and 0.80 ± 0.04 kg day⁻¹, respectively (P = 0.03).

Conclusions

Spring-born FGS suckler beef calves require minimal anthelmintic treatment to maintain performance. In contrast, clinical parasitic disease may develop in the SGS unless appropriate anthelmintic treatment is provided.     

The efficacy of two isolates of the nematode-trapping fungus Duddingtonia flagrans against Dictyocaulus viviparus larvae in faeces.

A series of experiments was carried out to examine the effects of two different isolates of the nematode-trapping fungus Duddingtonia flagrans to reduce the number of free-living larvae of the bovine lungworm, Dictyocaulus viviparus. A laboratory dose-titration assay showed that isolates CI3 and Troll A of D. flagrans significantly reduced (P < 0.05 to P < 0.001) the number of infective D. viviparus larvae in cultures at dose-levels of 6250 and 12,500 chlamydospores/g of faeces. The larval reduction capacity was significantly higher for Troll A compared to CI3 when lungworm larvae were mixed in faecal cultures with eggs of Cooperia oncophora or Ostertagia ostertagi and treated with 6250 chlamydospores/g of faeces. Both fungal isolates showed a stronger effect on gastrointestinal larvae than on lungworm larvae. Two plot trials conducted in 1996 and 1997 involved deposition of artificial faecal pats containing free-living stages of D. viviparus and C. oncophora on grass plots. Herbage around the pats was collected at regular intervals and infective larvae recovered, counted and identified. These experiments showed that both D. flagrans isolates reduced the number of gastrointestinal as well as lungworm larvae in faecal pats. During both plot trials, the transmission of C. oncophora larvae, but not D. viviparus, from faecal pats to the surrounding herbage was clearly affected by climatic conditions. After collection of faecal pats from the grass plots one month after deposition, the wet and dry weight of pats as well as organic matter content were determined. No differences were found between the fungus-treated and non-treated control pats. This indicated that the rate of degradation of faeces was not affected by the addition of the fungus.     

Chemotherapeutic efficacy of sulphadimidine, amprolium, halofuginone and chloroquine phosphate in experimental Eimeria bareillyi coccidiosis of buffaloes

Sulphadimidine, amprolium, halofuginone and chloroquine phosphate were administered to buffalo calves 10 days after experimental infection with Eimeria bareillyi. Animals given sulphadimidine or amprolium remained clinically normal and shed only a few oocysts in their faeces. Halofuginone was found partially effective and chloroquine phosphate completely ineffective in preventing faecal oocyst discharge and intestinal lesions. Sulphadimidine and amprolium treated calves gained weight, but chloroquine treated calves suffered progressive weight loss similar to that of infected untreated controls. No significant alterations of haematological values were observed either in the treated calves or in the untreated controls.     

Isolation of infective Dictyocaulus larvae from herbage

A technique is described in which Dictyocaulus larvae are stimulated to activity and isolated free from any debris by migration in agar gel.Microscopical recognition and enumeration was carried out on stained parasitic larvae while the free living species were decolourized.The technique was found suitable for routine use. Its efficiency was tested by adding larvae to the debris from parasite free herbage samples. Average recovery rates for Dictyocaulus viviparus and D. filaria larvae were found to be 60 and 80% respectively.     

Administration of Bacillus coagulans in calves: recovery from faecal samples and evaluation of functional aspects of spores

An investigation was carried out into the recovery from calf faeces of Bacillus coagulans spores added to the feed as probiotic. For this purpose, Bacillus coagulans spores (9 log₁₀ CFU g^?1) were given daily to 10 calves during the whole farming periods; another 10 calves acted as controls. Throughout the trial the faecal spore counts were significantly (P?<?0.01) higher in the treated group than in the controls (averaging 2.1?×?10⁵ vs 3.7?×?10⁴?CFU g^?1). Bacterial cells were recovered from faecal samples and ribotyping matched the strain isolated from faecal sample to the clone administered to the animals. In addition, the recovered cells were found to maintain their functionality aspects of acid production, survival in artificial gastric juice and in the presence of bile, and attachment to human intestinal epithelial cells. The results further elucidate the fate of spore formers administered to calves, and this will help in the development of new species-specific nutritional strategies.     

Development of free-living stages of equine strongyles in faeces on pasture in a tropical environment

The development of the free-living stages and yields of infective third stage strongyle larvae in faeces from a horse with a mixed natural infection deposited on pasture plots were studied over a 2-year period in a coastal area in tropical north Queensland. Two sets of faecal masses (one exposed to, and the other protected from the action of a natural population of dung beetles) were deposited monthly and after 7 days faecal samples were taken for larval recovery and counts. Hatching and development of the free-living stages occurred in faeces on pasture throughout the year. Development was rapid as infective stages were reached within a week of faecal deposition in all months. Yields of infective larvae were affected by the season and the action of dung beetles on the faecal masses. Highest yields were obtained from both beetle-exposed and protected faeces during winter (June to August) and lowest yields were in spring (September to November). High temperatures in spring and summer resulted in low yields of larvae, however, the dry conditions in spring made this season the most unfavourable period. In autumn and winter the temperatures were never low enough to stop or markedly slow down the rate of development, and allowed the development of large numbers of infective larvae. Dung beetle activity was observed throughout the year, and exposed faeces were usually completely dispersed within 24 h of deposition. This resulted in lower yields of infective larvae from these than from protected faeces. Though larval yields were lower, the actual numbers were still substantial so as to cast doubt on the usefulness of these beetles as biological control agents for equine strongylosis in the dry tropics.     

Seasonal translation of infective larvae of gastrointestinal nematodes of cattle and the effect of Duddingtonia flagrans: a 3-year pilot study

A study was conducted over 3 years (1998-2000) to investigate larval availability of gastrointestinal nematodes from faeces of cattle reared under different parasite control schemes. These cattle were part of a parallel, but separate grazing trial, and were used as donor animals for the faecal material used in this experiment. At monthly intervals, faeces were collected and pooled from three groups of first-season grazing cattle. These groups were either untreated, ivermectin bolus treated or fed the nematophagous fungus Duddingtonia flagrans. The untreated and fungus treated animals were infected with gastrointestinal nematodes and the number of eggs per gram (epg) pooled faeces ranged between 50 and 700 in the untreated group and between 25 and 525 epg in the fungus treated group. Each year between June and September, artificial 1 kg dung pats were prepared and deposited on pasture and protected from birds. The same treatments, deposition times and locations were repeated throughout the study. Larval recovery from herbage of an entire circular area surrounding the dung pats was made in a sequential fashion. This was achieved by clipping samples in replicate 1/4 sectors around the dung pats 4, 6, 8 and 10 weeks after deposition. In addition, coinciding with the usual time of livestock turn-out in early May of the following year, grass samples were taken from a circular area centred where the dung pats had been located to estimate the number of overwintered larvae, which had not been harvested during the intensive grass sampling the previous year. It was found that recovery and number of infective larvae varied considerably within and between seasons. Although the faecal egg counts in 1999 never exceeded 300 epg of the faecal pats derived from the untreated animals, the abnormally dry conditions of this year generated the highest level of overwintered larvae found on herbage in early May 2000, for the 3 years of the study. Overall, biological control with D. flagrans significantly reduced larval availability on herbage, both during and between the grazing seasons, when compared with the untreated control. However, the fungus did not significantly reduce overwintered larvae derived from early season depositions (June and July), particularly when dung pats disappeared within 2 weeks after deposition. Very low number of larvae (<3 per kg dry herbage) were sporadically recovered from grass samples surrounding the ivermectin bolus faecal pats.     

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

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

Ectonucleotidase and adenosine deaminase as inflammatory marker in dairy cows naturally infected by Dictyocaulus viviparus

The aim of this study was to evaluate the influence of dictyocaulosis (mild or severe) on enzymes of NTPDase, 5′-nucleotidase, and adenosine deaminase (ADA) of dairy cows naturally infected by Dictyocaulus viviparus. Blood and faeces were collected from 22 dairy cows of the same farm to evaluate NTPDase (ATP and ADP substrate), 5′-nucleotidase, and ADA activities on days 0 (pre-treatment) and 10 (post-treatment). Seric activities of NTPDase (ATP substrate), 5′-nucleotidase, and ADA were lower (P < 0.05) in D. viviparus infected animals compared to uninfected cows. The number of D. viviparus larvae per gram of faeces varied among the animals, and they showed different degrees of severity according to respiratory clinical signs of the disease (cough and nasal discharge). Later, these cows were divided into two groups: those with mild (n = 10) and severe (n = 12) disease. Cows with severe disease showed higher NTPDase activity (ATP substrate) than those with mild disease (P ≤ 0.05). The opposite occurred with NTPDase (ADP substrate), 5′-nucleotidase, and ADA in cows with severe disease, that is, the enzymatic activity of these seric enzymes significantly decreased (P ≤ 0.05) compared to animals with mild disease. Infected animals showed reduced NTPDase activity (ATP and ADP substrate) after treatment. No enzymatic changes were observed for 5′-nucleotidase, and ADA pre- and post-treatment (P > 0.05). Based on these results, we conclude that dictyocaulosis alters NTPDase, 5′-nucleotidase, and ADA activities of cow naturally infected by the parasite, in consequence the enzymes act as inflammatory markers.     

Effect of different times of administration of the nematophagous fungus Duddingtonia flagrans on the transmission of ovine parasitic nematodes on pasture--a plot study

Investigations were made into the timing of administration of Duddingtonia flagrans as a biological control agent against ovine parasitic nematodes including stongylid and Nematodirus spp. Faeces from 3-4 months old male lambs were deposited onto pasture plots that had never been grazed by sheep. The trial was conducted over two consecutive years (1998 and 1999). For both years, the following three plot types were involved: Sim plots had faeces containing nematode eggs and Duddingtonia flagrans spores deposited simultaneously; Post plots had faeces containing nematode eggs followed 2 weeks later by faeces containing D. flagrans spores alone; Control plots had faeces containing only nematode eggs; Prior plots (included in 1999) had faeces containing D. flagrans spores alone followed 2 weeks later by faeces containing nematode eggs. In each year, two deposition periods were involved: July and August in 1998 and June and July in 1999. During the first year pasture samples were collected at 2, 4, 6, 8 and 12 weeks after initial deposition. In 1999, additional samples were collected at 10, 16 and 20 weeks. Larvae were extracted from the pasture samples and counts performed to estimate the number and species of infective third-stage (L(3), larvae) present. The number of third-stage strongylid larvae on pasture was significantly lower on Sim plots compared to the remaining plot types for both years at all deposition times (P<0.001). This was also the case for the number of Nematodirus infective larvae in August deposition plots in 1998 (P<0. 02). There was no significant difference between treatments in both deposition times in 1999 and July deposition plots in 1998 for the Nematodirus data. These results suggest that D. flagrans, if deposited at the same time as parasite eggs prevents transmission of third-stage larvae from the faecal deposit onto pasture, including occasionally Nematodirus species, but does not have an effect on third-stage parasitic nematode larvae in the surrounding soil.     

Globule Leukocytes and bovine Parasitic Bronchitis

Globule leucocytes (GL) occur in the respiratory tract of cattle from foetuses to adult animals. Large numbers of this cell have been found in the lungs of cows both in normal lung and in the lungs of cows from various outbreaks of respiratory diseases, but the significance was not known. In the rat and in cattle, the occurrence of GL in the gastrointestinal and urinary tracts has been associated with parasitic infections. The relationship between the respiratory tract GL and Dictyocaulus viviparus infections in cattle was studied in three groups of parasite free calves experimentally infected with D. viviparus. Group 1 animals received single challenge infections. Group 2 had three subgroups (2a, 2b, 2c) which were either vaccinated and challenged (2a), given a single challenge (2b) or used as vaccinated controls (2c). Group 3 with four subroups (3a, 3b, 3c, 3d) were either used as environmental controls (3a), vaccinated and repeatedly challenged (3b), given normal larvae of D. viviparus and repeatedly challenged (3 c) or as a trickled group 3 d. Although GL were found in infected calves, it was not possible to demonstrate a clear cut correlation between their presence or density and the method of exposure.