Attempts to control infection with Ostertagia ostertagi (Trichostrongylidae) in grazing calves by adding mycelium of the nematode-trapping fungus Arthrobotrys oligospora (Hyphomycetales) to cow pats

Artificially prepared cow pats containing Ostertagia ostertagi eggs were deposited on two comparable grazing plots at weekly intervals during June and July 1987. Before deposition the cow pats, on the one plot, were inoculated with 0.250 g mycelial fragments per kg of faeces of the predacious fungus Arthrobotrys oligospora. On the other control plot comparable numbers of non-inoculated cow pats were placed at the same time. Subsequently, in July, two equal groups of calves were turned out one on each plot to graze for two months after which they were necropsied. The number of infective O. ostertagi larvae in inoculated cow pats was reduced by 42% and herbage larval infectivity around them by 50-71% as compared with the corresponding parameters in control pats and surrounding herbage. After a grazing period of two months the calves on the plot with inoculated cow pats had acquired worm burdens that were on average 37% lower than those of calves on the control plot. The lower degree of parasitism in these calves was also reflected in lower egg counts, lower serum pepsinogen levels and higher body weight.     

Field experiments on the ability of Arthrobotrys oligospora (Hyphomycetales) to reduce the number of larvae of Cooperia oncophora (Trichostrongylidae) in cow pats and surrounding grass

In field experiments, conducted on parasite-free grass plots in two consecutive summers, artificially prepared cow pats containing Cooperia oncophora eggs were inoculated with the nematode-trapping fungus Arthrobotrys oligospora. Numbers of infective C. oncophora larvae isolated from the pats as well as from the surrounding herbage were subject to an approximately ten-fold reduction as compared with numbers in fungus-free pats and herbage surrounding these. This reduction was undoubtedly a result of entrapment of the parasite larvae within the faecal pats.     

Induction of Nematode-Trapping Organs in the Predacious Fungus Arthrobotrys oligospora (Hyphomycetales) by Infective Larvae of Ostertagia ostertagi (Trichostrongylidae)

Laboratory experiments were designed to study the influence of temperature, concentrations of nematodes, oxygen tension, light, and nutrient levels, on the induction of nematode-trapping hyphal nets in the predacious fungus Arthrobotrys oligospora. When induced by infective Ostertagia ostertagi larvae, a maximum number of nets was produced at 20°C, at which temperature nets in surplus were produced at larval concentrations up to 1,000 larvae per cm2. A. oligospora did not produce nets in an anaerobic atmosphere containing 21 % CO2 (v/v), and net induction was suppressed to a certain degree by exposure to light. The composition of the medium had an important influence on the saprophytic growth and the net-forming capability of A. oligospora as a maximum number of nets was induced at a relatively low concentration of corn meal supporting the relatively sparse mycelium. It was shown that a proportion of trapping nets in A. oligospora maintained their trapping potential for more than 7 weeks when the temperature was below 25°C. Induction of nematodetrapping organs in A. oligospora is discussed in relation to control of infective nematode parasite larvae in cow pats.     

Field experiment on the ability of earthworms (Lumbricidae) to reduce the transmission of infective larvae of Cooperia oncophora (Trichostrongylidae) from cow pats to grass

In Denmark earthworms play an important role in the disappearance of cow pats on pastures. Field investigations were designed to study the influence of earthworms on the transmission of infective Cooperia oncophora larvae from experimental cow pats to grass. Results from the present experiment showed that cow pats, protected from attack by earthworms, disappeared at a much lower rate than unprotected cow pats. Development in faeces and transmission to grass of C. oncophora larvae were disturbed by the disintegration of cow pats. The rapid disintegration of unprotected cow pats resulted in an approximately 50% reduction of infective C. oncophora larvae on grass in the vicinity of these cow pats, as compared with larval contamination of grass around protected cow pats. How earthworms may cause a reduction in the transmission of infective C. oncophora larvae to grass by creating adverse conditions for the survival of parasite larvae in the cow pats, by eating and killing the parasite larvae, and by depositing parasite larvae in the soil is discussed.     

The predatory capability of three nematophagous fungi in the control of Haemonchus contortus infective larvae in ovine faeces

The effect of oral administration of three different nematode-trapping fungi, in aqueous suspension containing either Dactylaria sp. or Arthrobotrys oligospora conidia or Duddingtonia flagrans chlamydospores, on the number of Haemonchus contortus infective larvae in sheep faeces, was evaluated. The three selected species of fungi produce three-dimensional adhesive nets in the presence of nematodes. Sixteen Creole sheep were divided into four groups of four animals each. Groups 1 and 2 were orally drenched with a suspension containing 2x10(7) conidia of either A. oligospora or Dactylaria sp. Group 3, received a similar treatment, with D. flagrans chlamydospores, instead of conidia, being administered, at the same dose. Group 4 acted as control, without any fungi. Faecal samples were collected directly from the rectum of each sheep and faecal cultures were prepared and incubated at 15 and 21 days. Larvae were recovered from faecal cultures and counted. The highest reduction of the nematode population occurred in the D. flagrans group, reaching reductions of 96.3% and 91.4% in individual samplings in plates incubated for 15 and 21 days, respectively. Arthrobotrys oligospora showed moderate reductions in the faecal larval population, ranging between 25-64% at 15 days incubation. In general, Dactylaria sp., was less efficient in its trapping ability. Despite the inconsistent results with Dactylaria sp., reduction percentages of 73.4% and 80.7% were recorded in individual samplings during the first and second days, in plates incubated for 15 days. Duddingtonia flagrans, was shown to be a potential biological control agent of H. contortus infective larvae.     

Ecology of the free living stages of cattle nematodes during summer contamination in Argentina Western Pampas

Development, migration and survival of infective larvae (L3) were studied in the Western Pampeana Region. Faeces of naturally nematode infected cattle were deposited as artificial pats on plots during mid-spring-summer of 1994/1995 and 1995/1996. Since the start and during 1995, the study coincided with a severe drought, rainfalls being 29% below the 45-year means. The predominant genera recovered were Cooperia, Ostertagia and Haemonchus. Initial and peak recovery of L3 from pats occurred 8-15 and 15-21 days later respectively. A low percentage of L3 survived from November (0.3% L3) and January (0.06% L3) to the following autumn and winter. The mean persistence of larvae detected in pats or herbage was around 200 days from deposition. The migration of L3 from faecal pats to herbage started 15 to 30 days after deposition according to rainfall occurrence. Maximum herbage recoveries of L3 from pats deposited in late summer occur during autumn rainfalls. Only, few L3 were occasionally recovered from soil. Summer conditions were associated with rapid development and translation of L3 to herbage, but also with low L3 detection after initial recoveries. Faecal pats deposited from mid-summer were the main source of autumn herbage contamination.     

The influence of weather and egg contamination on the development of third-stage larvae of Cooperia oncophora on pasture.

The influence of weather and egg contamination on the dynamics of herbage contamination with infective larave of Cooperia oncophora was investigated on artificially contaminated grass plots and in a grazing experiment with 24 first-year grazing calves from May to October 1987 in Lower Saxony, Germany. On the experimental plots the larval translation was highest at the beginning of July and in the second part of September, following high mean weekly temperatures. Between July and September peak recovery of larvae from herbage occurred 4 weeks after contamination. A seasonal pattern of larval translation similar to that on the experimental plots could be demonstrated on the grazed pastures when the number of larvae per m2 of pasture had been adjusted to the previous egg output by means of a contamination index. The resulting 'relative larval density' is regarded as a good indicator for larval development on pasture. From July to September the larval population on pasture resulted mainly from the egg contamination 2-3 weeks earlier. The short persistence of the infective larvae on herbage was probably due to the frequent and heavy rainfall throughout the season, causing a passive washout of larvae into the soil. On single pastures the larval density started to increase within 1 week after the calves had first contact with these fields. The impact of the calves on the distribution of larvae is discussed.     

In vivo passage through calves of nematophagous fungi selected for biocontrol of parasitic nematodes.

The experiment was designed to test the survival and performance of stress selected nematophagous fungi after passage through the gastro-intestinal tract of cattle. Ruminating calves were fed daily with a fixed amount of fungal material grown on barley grains. The excreted dung was collected on days four and five after the start of the feeding experiment. Barley grains were washed out of the excreted dung and incoculated on water-agar plates. After incubation for one week, nine of ten fungal isolates were re-isolated from these plates. The predatory capacity of the fungi in the excreted faeces was tested in a dung pat bioassay and a faecal culture system. In the dung pat bioassay, two fungi of the genus Arthrobotrys and six of the genus Duddingtonia reduced the development of Ostertagia ostertagi third stage larvae by 85% (61%-93%), compared to the number of larvae developed from fungus-free control pats. In seven out of these eight isolates, the reduction of larvae in the faecal cultures was 92% (76%-99%).     

Formulation of a strategy for the application of Duddingtonia flagrans to control caprine parasitic gastroenteritis

Experiments on the influence of egg density and varying quantities of chlamydospores on the nematode-trapping ability of Duddingtonia flagrans, influence of D. flagrans on the larval translation of gastrointestinal nematodes, doses of chlamydospores required for the effective control of gastrointestinal nematodosis and the epidemiology of nematode parasites were conducted in goats, which generated baseline data required for strategic application of the biocontrol agent. The nematode-trapping ability of D. flagrans, measured by numerical enumeration of infective third-stage larvae developed in the faecal culture, revealed that the efficacy is dependent on both nematode egg and chlamydospore density. Pasture plot studies revealed that D. flagrans, if deposited at the same time as nematode eggs, prevents translation of third-stage larvae of caprine nematodes from the faecal pats onto the grass blades. Feeding of 1 x 106 chlamydospores per kg body weight and above to goats virtually eliminated larvae from the pasture. Application of as few as 1 x 104 and 1 x 105 chlamydospores per kg body weight had a profound impact on larval recovery. The effect persisted as long as the chlamydospores were fed. Monthly faecal worm egg counts of adult goats maintained under a semi-intensive management system on the Chhattisgarh plain and pasture larval burden revealed that June to August were the months of high risk for nematodosis. Haemonchus was the dominant species recorded throughout the year. The present data can best be utilized by formulating a strategic control measure when the larval challenge to the animal is maximum (June to August), so as to prevent establishment of patent infection. The observations reinforced the strategy to be adopted for nematode parasite control in goats by applying the biocontrol option at the onset of the monsoon.     

A Field Experiment on Rain Splash Dispersal of Infective Larvae of Ostertagia Ostertagi (Trichostrongylidae) from Cow Pats to Surrounding Grass

Ostertagia ostertagi is an important tricho-strongyle nematode parasite of cattle in several countries, including Denmark. It leaves its host as eggs with faeces. After hatching it develops in the cow pat up to the third and infective stage, which is then transmitted to the surrounding grass.     

Pasture populations of cattle nematode larvae in Guadeloupe (French West Indies)

The development of numbers of third-stage (L3) larvae of gastrointestinal nematodes after pat (faeces) deposition by heifers is described, using a mathematical model, for seven experiments carried out in Guadeloupe (French West Indies). A dramatic rise in L3 population size occurred on herbage near pats, 17-25 days after pat deposition. There was no clear relationship between climatic data and L3 population dynamics. However, 5 successive days of rainfall induced the resumption of larval migration in two experiments. Changing the unit of expression of L3 population size (100 L3 m-2 vs 1000 L3 per kg DM) did not alter estimates of the model parameters. The evolution of L3 population of Haemonchus and Cooperia were similar.     

Seasonal variations in the populations of infective larvae on pasture and the numbers of nematode eggs in the faeces of farmed goats

The levels of nematode egg production in goats and the availability of infective larvae (L3) on pasture were investigated on a dairy unit in New South Wales, Australia. The output of eggs by adult goats was always above 300 epg. The profile of the graph of larval availability in herbage paralleled those for temperature and rainfall, suggesting that larval peaks occurred when the temperature and availability of moisture were optimal. The dominant genus was Trichostrongylus, followed by Haemonchus, then Ostertagia. A larger proportion of Haemonchus larvae in the cultures of faeces were collected during the summer months.     

Pathogenicity of Paecilomyces fumosoroseus isolates to diamondback moth, Plutella xylostella: correlation with spore size, germination speed, and attachment to cuticle

Infectivity to larvae of the diamondback moth, Plutella xylostella, was compared among eight Paecilomyces fumosoroseus isolates. Isolate infectivity was assessed for correlation with spore length and germination speed. Four isolates applied to P. xylostella cuticle were also compared for number of spores remaining on the cuticle after washing and for percentage germination after 36 h. Infection of larvae inoculated with the different isolates at an average dosage of 4000 conidia/cm2 ranged from 2 to 47%. The correlation of infectivity with spore length and germination speed in broth was highly significant. Fewer spores of the least infective isolate, ARSEF 1576, attached to larval cuticle compared to spores of the more infective isolates ARSEF 3682, 4461, and 4482 (P < 0.05). After 36 h on larval cuticle, the percentage of spores germinated for isolates 1576 and 3682 was 3 and 95%, respectively. Spores of isolate 1576 were smaller, germinated more slowly, and attached to cuticle in smaller numbers than spores of the more infective isolates. Further research will expand our understanding of the mechanisms of virulence among isolates of P. fumosoroseus.     

The ecology of infective larvae of bovine gastrointestinal trichostrongylids in dry season contaminated pastures in the Nigerian derived savanna

Five experimental grass paddocks were sequentially contaminated with fresh bovine faeces containing known numbers of eggs of predominantly Haemonchus and Cooperia spp. during the 1984/85 dry season (November to March). Faecal, herbage and soil samples were examined at regular intervals between November and June in order to determine the rate of development and mortality of infective larvae (L3) in faeces, the pattern of herbage infestation with L3 and the role of faeces and soil as reservoirs of L3 during the dry season and early rains. L3 first appeared in faeces approximately 4 days post contamination (PC) and peak counts were obtained 28, 14, 7 and 14 days PC in the paddocks contaminated in December, January, February and March, respectively. The counts initially declined linearly at the rate of approximately 7535 L3/week and 10,947 L3/week in P2 and P4 respectively, due primarily to mortality but later there was an accelerated fall in the counts as the surviving L3 moved out of the faecal pads onto herbage. The overall trend of faecal larval populations in each paddock was therefore distinctly curvilinear. Although large numbers of L3 were present inside dry faecal pads in most paddocks throughout the dry season, none migrated on to herbage at that time of the year. Translation of L3 to herbage was very rapid and occurred simultaneously in all the paddocks 24 hours following the first heavy rainfall in late March. Consequently peak herbage infestations in all paddocks were coincident and occurred a few days after commencement of larval migration. The closer to the end of the dry season the contaminations were carried out, the larger were the subsequent early rains rise and the peak herbage infestation and the longer this infestation survived on herbage. No L3 were recovered from soil throughout the study, which suggests that faecal pads were the sole reservoir of L3 during the dry season and hence the source of the early rains herbage infestation.     

Epizootiology of a nuclear-polyhedrosis virus in European spruce sawfly,Gilpinia hercyniae: Birds as dispersal agents of the virus during winter

In Welsh spruce forest the relationship of birds to a nuclear polyhdedrosis virus (NPV) was studied outside the larval period of its sawfly host, Gilpinia hercyniae. Birds passed feces containing infective NPV throughout the nonlarval period (November–June). It is thought that birds acquired inoculum by feeding on the corpses of NPV-killed larvae adhering to spruce trees. The proportion of infective droppings declined in the 2 months (May and June) prior to the new larval period. Infective droppings were collected from all six species of birds netted in January. The infectivity of droppings of goldcrest was less and that of longtailed tit greater in January than in the previous September. Experimental evidence suggests that birds may carry NPV for at least 6 km.     

A Study on the Transmission of Oesophagostomum dentatum and Hyostrongylus rubidus among Outdoor Reared Pigs in Denmark

This study was carried out to obtain basic information on the transmission of Oesophagostomum dentatum and Hyostrongylus rubidus in outdoor reared pigs in Denmark. Eighteen 10 weeks old worm-free pigs were allocated into 3 groups of 6 pigs each. In May, all pigs were turned out on the same parasitologi-cally naive pasture, and after 2 weeks the pigs in groups 2 and 3 were experimentally infected with 10,800 O. dentatum and 8,700 H. rubidus infective larvae, respectively. Pigs in group 1 served as non-infected controls. All pigs were reared together on the experimental pasture for further 134 days until slaughter in October. Strongyle egg counts, differentiation of infective larvae at species level, serum pepsinogen, and herbage larval infectivity were monitored at regular intervals throughout. Both strongyle species established in the originally parasite-free pigs (group 1) and cross infections were established in group 2 and 3. The pigs were exposed to steadily increasing herbage infectivity of both species of strongyles. At the end of the experiment, geometric mean worm burdens of O. dentatum in groups 1,2 and 3 were 1202, 6136 and 1431 respectively, the burden in group 2 being significantly higher (p<0.05) than that of the 2 other groups. The geometric mean worm burdens of H. rubidus in groups 1, 2 and 3 were 4907, 3679 and 5246 respectively, showing no significant differences between groups.     

Survival of Campylobacter spp. in bovine faeces on pasture

Aims:  To determine the survival on pasture of Campylobacter spp. naturally present in bovine faeces and compare this with a previously published study using laboratory-cultured Campylobacter spp.
Methods and Results:  Ten freshly collected cow pats were deposited on pasture during summer, and Campylobacter spp. were enumerated by enrichment broth culture. The counts in three pats were below detection limits. Counts of Campylobacter spp. in the other seven pats fell below detection limits within 14 days. The geometric means of the counts up to 7 days produced a T ₉₀ of 2·2 days. Characterization of Campylobacter spp. by PCR and pulsed field gel electrophoresis indicated the presence of at least six genotypes of Campylobacter jejuni , Campylobacter coli and Campylobacter lari .
Conclusions:  Campylobacter spp. naturally present in cow faeces exhibited a similar survival rate to that previously determined using laboratory-cultured strains. The highly variable counts of naturally occurring Campylobacter spp., and the predominance of lower counts, also support the earlier decision to use laboratory-cultured strains in survival experiments.
Significance and Impact of the Study:  This study reaffirms the short survival of Campylobacter spp. in cow faeces deposited on pasture. This information will be incorporated into a 'reservoir model' for Campylobacter spp. in cow pats on New Zealand pastures.     

Factors influencing the infectivity of isolates of Paecilomyces fumosoroseus against diamondback moth, Plutella xylostella

Paecilomyces fumosoroseus isolate 1576 was isolated from an insect, but is avirulent against larvae of diamondback moth, Plutella xylostella, and several other species. Isolate 1576 grew faster and produced more conidia than isolate 4461 on potato dextrose agar. Pregermination of conidia failed to increase the infectivity of isolate 1576, but the procedure did increase the infectivity of isolates 3682, 4461, and 4482. Isolates 1576 and 4461 were both more infective when moisture was high during incubation of inoculated larvae. Starved Pl. xylostella larvae were more susceptible than fed larvae to isolate 1576 (40 and 10% mortality, respectively), but starved and fed larvae were similar in susceptibility to isolate 4461. These results show that isolate 1576 grows vigorously in aerial culture and is capable of infecting stressed Pl. xylostella larvae. Further tests are needed to characterize its pathogenicity toward its original host or closely related species.     

The roles of ascospores and conidia of Pyrenopeziza brassicae in light leaf spot epidemics on winter oilseed rape (Brassica napus) in the UK

Ascospores of Pyrenopeziza brassicae were produced in apothecia (cup‐shaped ascomata) on oilseed rape debris. The conidia, which were morphologically identical to the ascospores, were produced in acervular conidiomata was greater than for lesions caused by ascospores. In June 2000, on the ground under a crop with light on the surface of living oilseed rape tissues. Ascospores were more infective than conidia on oilseed rape leaves. The proportion of lesions caused by conidia located on leaf veins leaf spot, numbers of petioles with apothecia decreased with increasing distance into the crop from the edge of pathways. Air‐borne ascospores of P. brassicae were first collected above debris of oilseed rape affected with light leaf spot on 5 October 1998 and 18 September 1999,12 or 23 days, respectively, after the debris had been exposed outdoors. P. brassicae conidia were first observed on leaves of winter oilseed rape on 6 January 1999 and 15 February 2000, respectively, after plots had been inoculated with debris in November 1998 and October 1999. In 1991/92, numbers of ascospores above a naturally infected crop were small from January to April and increased in June and July. P. brassicae conidia were first observed in February and the percentage plants with leaves, stems or pods with light leaf spot increased greatly in May and June. In 1992/93, in a crop inoculated with debris, numbers of airborne ascospores were small from October to January and increased from April to June. P. brassicae conidia were first observed on leaves in late November and light leaf spot was seen on stems and pods in March and June 1993, respectively.     

Laboratory experiments to evaluate the ability of Arthrobotrys oligospora to destroy infective larvae of Cooperia species, and to investigate the effect of physical factors on the growth of the fungus

Laboratory investigations were designed to study the influence of temperature, pH and oxygen tension on the growth of Arthrobotrys oligospora, a nematode-trapping microfungus. Experiments were performed to evaluate the potential role of A. oligospora in destroying third-stage larvae of Cooperia spp. on agar plates and in cattle faeces. The fungus had a growth rate optimum at 23 degrees C and pH 6. Anaerobic cultivation for 23 hours at 23 degrees C and 39 degrees C inhibited fungal growth, but it did not destroy the fungus, which regained growth upon a subsequent shift to aerobic conditions at 23 degrees C. Under experimental conditions in petri-dishes containing agar, the nematode-trapping efficiency of the fungus was striking in that 100% of a population of third-stage larvae of Cooperia spp. was captured within three days of the experiment. The trapping efficiency in faeces was shown to depend upon the inoculation level. At a concentration of approximately 2500 conidia per g faeces, 99% of the larvae were destroyed. The possibilities of using nematode-trapping fungi in controlling animal-parasitic nematodes are discussed.