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.     

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.     

The ecology of the free-living stages of Ostertagia circumcincta

Callinan A. P. L. 1978. The ecology of the free-living stages of Ostertagia circumcincta. Internationaljournal for Parasitology8: 233–237. The development and survival of the free-living stages O. circumcincta was studied in western Victoria in 1974–1976. For all plots in which development occurred, pre-infective larvae (L₁-L₂) were recovered within 0–8 days, infective larvae (L₃) within 7–30 days and L₃ on herbage and soil within 4–27 days. The mean minimum development time to L₃ on herbage and soil was 14·3 ± 3·5 days and the mean development time to maximum yield of these was 33·3 ± 6·3 days. There were no distinct differences in development rates between plots. Plot yields of L₃ on herbage and soil varied inversely as the mean of daily temperatures for the period until maximum yield of these. Yields varied from 0 to 16% of the number of eggs put out on each plot and highest yields were obtained from eggs put out during late autumn-winter. No L₃ were observed to survive over summer. The migratory habits of L₃ were such that a mean of 75·1 ± 5·6% of L₃ on herbage and soil were actually on herbage; the soil was always a significant source of L₃.     

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.     

Effect of plant species on the larvae of gastrointestinal nematodes which parasitise sheep

Faeces containing Trichostrongylus colubriformis and/or Ostertagia circumcincta eggs were used to provide four contaminations in each of 2 years on plots of browntop, Yorkshire fog, ryegrass, tall fescue, lucerne, chicory, cocksfoot, white clover, and prairie grass and in the second year a mixed sward of ryegrass/white clover. Third stage larvae were recovered from faeces and from four strata of herbage, 0–2.5, 2.5–5, 5–7.5 and >7.5 cm above the soil surface at 2, 4, 6, 8, 11, and 14 weeks after faeces were deposited on the swards. Herbage species had a significant (P < 0.0001) effect on the number of larvae recovered. Greatest numbers of larvae, as indicated by ranking analysis, were recovered from Yorkshire fog, ryegrass, and cocksfoot and lowest numbers from white clover and lucerne. The difference between herbages in numbers of larvae recovered was due to the ‘‘development success’’, the ability of larvae to develop to the infective stage and migrate on to herbage, rather than ‘‘survival’’, the rate of population decline once on the herbage. Faecal degradation was most rapid from white clover and browntop, intermediate from tall fescue, lucerne, prairie grass, cocksfoot, and ryegrass, and slowest from Yorkshire fog swards. The numbers of larvae recovered from herbages were related (r² = 0.59, P < 0.05) with the faecal mass remaining. A greater proportion of the total larvae recovered from the herbage was recovered from the bottom stratum of Yorkshire fog and prairie grass than from white clover, with the other herbages intermediate, indicating that larvae had greater difficulty migrating up Yorkshire fog and prairie grass than the other herbage species. In most herbage species, despite more larvae being recovered from the lowest stratum, larval density (L₃/kg herbage DM) was highest in the top stratum. This study has demonstrated that herbage species can have a significant impact on the population dynamics and vertical migration of T. colubriformis and O. circumcincta 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 development and overwintering survival of free-living larvae of Haemonchus contortus in Sweden

Five complimentary studies were undertaken with the overall aim to examine the ability of free-living stages of Haemonchus contortus to over-winter and tolerate cold stress. Two studies deal with the development and long-term survival of eggs and infective larvae of two geographically different isolates (Kenya and Sweden). Eggs and larvae were monitored in climatic chambers at temperatures that fluctuated daily between -1 degrees C and 15 degrees C, or at constant temperatures of 5 degrees C and 15 degrees C. The development from egg to larvae was dependent on temperatures over 5 degrees C. The long time survival was favoured at lower temperatures. Furthermore, the overwintering capacity of the free-living stages of these isolates was estimated under Swedish field conditions. Two groups of lambs were experimentally infected with different isolates, and kept separated on previously ungrazed plots. In early May the following year, two parasite-naive tracer lambs were turned out on each of the plots to estimate the pick up of overwintered larvae. This experiment was replicated in central and southern Sweden. In addition, two experiments were performed in 2003 on pasture previously grazed by naturally infected sheep. One trial was on a pasture in southern Sweden grazed by a commercial flock, where extreme numbers of H. contortus were found towards the end of the grazing season 2002. The other study was on a pasture plot in central Sweden grazed by a hobby flock in 2002, where three of six lambs died due to haemonchiasis. Overwintered H. contortus was recorded on three of four experimental sites. Worm burdens were in all instances extremely low. No differences in development and survival were found between the isolates. Consequently, overwintering on pasture is of no practical significance in the transmission of H. contortus between grazing-seasons in Sweden.     

Survival of Ancylostoma caninum on bluegrass pasture.

The survival of infective larvae of Ancylostoma caninum on outdoor grass plots was studied in 40 experiments over 1 year. Weather data were collected over the period. Mean larval survival from August to early November was 24 days (range 1 to 49), from December through February was 0 days, and from March to mid-August was 6.6 days (range 0 to 21). Moderate to high temperatures and substantial rainfall favored larval survival; low temperatures and rainfall favored larval destruction.     

The ecology of the free-living stages of Trichostrongylus axei

Callinan A.P.L. 1978. The ecology of the free-living stages of Trichostrongylus axei. International Journal for Parasitology8: 453–456. The development and survival of the free-living stages of Trichostrongylus axei was studied in western Victoria in 1974–1976. For all plots in which development occurred, preinfective larvae (L₁-L₂) were recovered within 0–5 days, infective larvae (L₃) in faeces within 4–28 days and L₃ on herbage and soil within 4–21 days. The mean minimum development time to L₃ on herbage and soil was 12.3 ± 0.7 days and the mean development time to maximum yield of these was 33.8 ± 7.4 days. A mean of 66.7 ± 6.6% of L₃ on herbage and soil were actually found on herbage. Yields of L₃ on herbage and soil varied from 0 to 8.9 % of the number of eggs put out on each plot. Yields varied approximately inversely as the mean daily temperatures for the period until maximum yield. No L₃ were observed to survive over summer.     

Survival and behaviour of unfed stages of the ticksRhipicephalus appendiculatus,Boophilus decoloratus andB. microplus under field conditions in Zimbabwe

The survival and behaviour of the unfed stages ofRhipicephalus appendiculatus, Boophilus decoloratus andB. microplus in gauze columns were observed in long and short grass in the highveld of Zimbabwe. Ticks were exposed in the cool, hot and rainy seasons of 1980 and 1981. All species and stages survived longer in long grass than in short grass. Larvae from engorged female ticks released in the cool season hatched much later than incubator-reared controls. They were consequently not present during the cold weather and survived longer than larvae subjected to the low temperatures, in which the shortest survival-times were recorded. The survival of nymphs was insensitive to season. The longest survival-times were recorded in adults. Median survival-times of incubator-reared adults ranged from 165 to 375 days in short grass and from 333 to 493 days in long grass. These times were usually longer than those for adults which moulted in the field. Larvae of the three species and nymphs ofR. appendiculatus were active soon after hatching or moulting, irrespective of the season. In contrast, adults ofR. appendiculatus showed different patterns of activity in different seasons. Adults first appeared at the base of the columns in October/November and then gradually ascended to reach a maximum height in December/January. They remained high up in the columns until May/June when the weather became increasingly cold and dry. Larvae ofB. decoloratus climbed higher up in the columns in the long grass than did the larvae of the other two species.Larvae and nymphs ofR. appendiculatus and larvae ofB. microplus migrated up and down the columns daily, but larvae ofB. decoloratus and adults ofR. appendiculatus did not migrate.     

Acquisition of freezing tolerance in early autumn and seasonal changes in gall water content influence inoculative freezing of gall fly larvae,Eurosta solidaginis (Diptera,Tephritidae)

We examined seasonal changes in freeze tolerance and the susceptibility of larvae of the gall fly, Eurosta solidaginis to inoculative freezing within the goldenrod gall (Solidago sp.). In late September, when the water content of the galls was high (approximately 55%), more than half of the larvae froze within their galls when held at -2.5 degrees C for 24 h, and nearly all larvae froze at -4 or -6 degrees C. At this time, most larvae survived freezing at > or = -4 degrees C. By October plants had senesced, and their water content had decreased to 33%. Correspondingly, the number of larvae that froze by inoculation at -4 and -6 degrees C also decreased, however the proportion of larvae that survived freezing increased markedly. Gall water content reached its lowest value (10%) in November, when few larvae froze during exposure to subzero temperatures > or = -6 degrees C. In winter, rain and melting snow transiently increased gall water content to values as high as 64% causing many larvae to freeze when exposed to temperatures as high as -4 degrees C. However, in the absence of precipitation, gall tissues dried and, as before, larvae were not likely to freeze by inoculation. Consequently, in nature larvae freeze earlier in the autumn and/or at higher temperatures than would be predicted based on the temperature of crystallization (T(c)) of isolated larvae. However, even in early September when environmental temperatures are relatively high, larvae exhibited limited levels of freezing tolerance sufficient to protect them if they did freeze.     

Influence of Chilling and Freezing Temperatures on Infectivity of Meloidogyne incognita and M. hapla

Egg masses and second-stage larvae of Meloidogyne incognita and M. hapla in soil were exposed to temperatures ranging from 20 to -8 C. Temperature was lowered in 2-day intervals to 16, 12, 8, 4, 0, -4, and -8 C, and the nematodes remained at 4, 0, -4, or -8 C for 18, 14, 10, or 6 days, respectively. Unhatched larvae of both species were more resistant to low temperatures than were embryonic stages. Within the eggs of M. incognita, 7.5% of embryos and 48% of larval stages survived 14 days at 0 C, whereas 9% of embryos and 90% of larval stages in the eggs of M. hapla survived 10 days at -4 C. Second-stage larvae of both species remained infective in sol.1 at 4 or 0 C, but were injured at -4 and -8 C. Infectivily of these larvae was lower in saturated soil than in soil at 51 cm moisture tension at all temperatures.     

Development and survival of infective larvae of Haemonchus contortus and Trichostrongylus colubriformis on pasture in a tropical environment

A trial to determine the seasonal pattern of egg hatching and larval survival on pasture was carried out in representative wet and dry zones of Fiji. Fourteen plots were established on parasite-free pasture at each of two sites. One plot at each site was contaminated every month with faeces from naturally infected goats containing a known proportion of Haemonchus contortus and Trichostrongylus colubriformis eggs. Pasture was sampled at regular intervals after contamination and infective larvae identified and counted. Larvae of both species developed throughout the year in the wet zone but development was more sporadic in the dry zone. Larval counts generally declined to below detectable levels within 9 weeks of contamination between September and March but longevity increased during the cooler weather from April to August. The comparatively short larval survival times noted in this experiment may present opportunities for manipulation of parasite population dynamics in the wet tropics.     

Cold-Hardiness and Supercooling Capacity in Diapausing and Nondiapausing Stages of the Cabbage Root Fly Delia radicum

Supercooling point (SCP) values and cold-hardiness were measured in individual ontogenetic stages of Delia radicum (Diptera:Anthomyiidae) in various physiological states (winter diapause, summer quiescence, and normal development). Winter diapause-destined mature third-instar larvae had a lower SCP (-9.9 degrees C) than their nondiapause counterparts (-5.2 degrees C), and more of them survived exposure to -10 degrees C for 5 h to pupariation and adult emergence. Values of SCPs were equal in both diapause and nondiapause states of prepupal and pupal stages. The lowest SCP (ca. -20 degrees C) was found in the stage of phanerocephalic pupa (PCP) regardless of the physiological state. The cold-hardiness of PCP corresponded with a low SCP value only in diapausing pupae stored for 80 days at 3 degrees C and in pupae which had terminated their diapause and whose further development was inhibited by storage at low temperatures (3 degrees C). Such pupae survived exposure to temperatures close to their SCP (14 days at -17 degrees C). However, this high cold-hardiness was only acquired after some time and/or exposure to 3 degrees C, as the PCP at the beginning of diapause showed significantly impaired cold-hardiness despite the fact that their SCP was low. The cold-hardiness of nondiapausing PCP did not correspond at all to that of low SCP, as no pupa survived the exposure to -17 degrees C for 1 day; survival rates at temperatures of -13.5 and -10 degrees C were also remarkably lower than those in diapausing pupae. Cold-hardiness in D. radicum was closely connected with the diapause syndrome but the changes in SCP value corresponded rather with the ontogeny of this insect. Copyright 1993, 1999 Academic Press.     

The survival of Ostertagia circumcincta and Trichostrongylus colubriformis in faecal culture as a source of bias in apportioning egg counts to worm species.

When cultured alone or concurrently with Trichostrongylus colubriformis in sheep faeces, Ostertagia circumcincta produced fewer infective larvae per 100 eggs than did T. colubriformis. Averaged over five trials 60% of T. colubriformis eggs were recovered as infective larvae while for O. circumcincta the figure was only 39%. This result was observed for two strains of O. circumcincta and was independent of when larvae were harvested from culture (days 6-10 at 25 degrees C). The mortalities of both species occurred at the first and second larval stages. These observations are of concern when using larval differentiation from faecal culture to make quantitative estimates of worm egg numbers for each species present. Species such as T. colubriformis which have a low mortality during culture are likely to have their egg numbers overestimated when cultured with a species, like O. circumcincta, that suffers high mortality in culture.     

Influence of Low Temperature on Rate of Development of Meloidogyne incognita and M. hapla Larvae

Development of Meloidogyne incognita and M. hapla larvae in clover roots was studied at 20, 16, 12, and 8 C in growth chambers and in the field from fall through spring, in North Carolina. Larvae of both species invaded roots and developed at 20, 16, and 12 C, but not at 8 C. The time necessary to complete the larval stages at each temperature was determined. The minimal temperature for development of M. incognita larvae was 10.08 C and 8.8 C for M. hapla larvae. In the field, soil temperature at 10 cm deep was favorable for development of larvae until the end of November, and again from February on. All stages of the nematodes survived freezing temperatures in the roots. Reproduction of both species was evident in March or Apri1 after inoculation and accumulation of 8,500 to 11,250 degree-hours.     

Adaptations to the Arctic: low-temperature development and cold tolerance in the free-living stages of a parasitic nematode from Svalbard

For nematodes with a direct life cycle, transmission is highly dependent on temperature-related development and survival of the free-living stages. Therefore, in the Arctic, where the winter lasts from October to May, nematode transmission is expected to be focused in the short summer season, yet there is strong evidence that as well as focussing egg output during winter months, the nematode parasite, Marshallagia marshalli, infects Svalbard reindeer during the Arctic winter when temperatures are persistently below freezing. To investigate the potential for development and survival of eggs and infective third-stage larvae in winter and therefore the possibility of for winter transmission, we ran a series of low-temperature laboratory experiments. These provide five key insights into the transmission and survival of the free-living stages of M. marshalli: (1) eggs hatched at temperatures as low as 2 °C, but not below 0 °C, (2) eggs were viable and developed after being exposed to sub-zero temperatures for up to 28 months, (3) infective-stage larvae survived for up to 80 days at 5 °C, (4) infective-stage larvae could survive rapid exposure to temperatures below ?30 °C, and (5) desiccation resistance may be important for long-term larval survival at low temperatures. Together, these results indicate that eggs deposited during the winter are highly tolerant of prevailing environmental conditions and have the potential for rapid development with the onset of spring. It is therefore likely that the parasite remains in the egg stage in the faeces during the winter of deposition, hatch and develop into the infective larval stage in the summer, remaining viable on the tundra until the reindeer host returns to the winter feeding grounds the following winter.     

Herbage response to tree thinning in a Eucalyptus crebra woodland

The effects of a range of tree densities on native herbage (mainly Aristida ramosa, Bothriochloa decipiens and Themeda australis biomass in a Eucalyptus crebra woodland near Kingaroy, Queensland, were investigated between March 1977 and July 1981. Rainfall in this area averages 750 mm year^?1. Initial tree density was 640 trees ha^?1 and this was manipulated using arboricide chemicals to leave plots containing 640, 320, 160, 80 and nil live trees ha^?1. Fires were excluded from the whole area, and half the plots were grazed by cattle. The largest increase in herbage biomass was recorded in the ‘all trees killed’ treatment (nil trees ha^?1), closely followed by the ‘scattered tree’ treatment (80 trees ha^?1). The relationship between tree density and herbage biomass was linear. Recruitment of grass and forb plants, as reflected by changes in density, varied according to treatment. Increased grass recruitment was correlated with cattle grazing, whilst forb recruitment was influenced mainly by tree density.     

Evaluating the effectiveness of a Mexican strain of Duddingtonia flagrans as a biological control agent against gastrointestinal nematodes in goat faeces

The use of Duddingtonia flagrans in the control of goat nematodes was investigated. Initially, the time of passage of chlamydospores through the digestive tract of goats was evaluated. Two groups of seven parasite-free kids were formed. Group A received a single dose of 3.5x10(6) D. flagrans chlamydospores (FTHO-8 strain) per kg of live weight. Group B did not receive any chlamydospores. Faeces were obtained from each kid daily from day 4 prior to inoculation until day 5 post-inoculation (PI) and were placed in Petri dishes containing water agar. Gastrointestinal nematode infective larvae were added to each Petri dish and incubated at 25 degrees C for 7 days. Petri dishes were examined to detect the fungus and trapped nematodes. A second trial evaluated the effect of D. flagrans on the number of gastrointestinal nematode larvae harvested from goat faecal cultures in naturally infected goats. Two groups of seven goats were formed. The treated group received a single dose of 3.5x10(6) D. flagrans chlamydospores per kg of liveweight. The control group did not receive any chlamydospores. Faeces were obtained twice daily from each kid. Two faecal cultures were made for each kid. One was incubated for 7 days and the other for 14 days. Gastrointestinal nematode larvae were recovered from each culture and counted. Percentage of larval development reduction was determined using a ratio of larvae/eggs deposited in the control and treated groups. Duddingtonia flagrans survived the digestive process of goats, and maintained its predatory activity, being observed from 21 to 81 h PI (3 to 4 days). A reduction in the infective larvae population in the treated group compared to the non-treated group was observed in both incubation periods (7 days: 5.3-36.0%; 14 days: 0-52.8%, P>0.05). Although a single inoculation of D. flagrans can induce a reduction of infective larvae collected from faeces, a different scheme of dosing may be needed to enhance the efficacy of D. flagrans in goats.     

Thermal requirements of Galleria mellonella L. (Lepidoptera: Pyralidae) immature stages

The rearing of Galleria mellonella L. in laboratory is important for multiplication of entomopathogenic nematodes, mandatory for biological control studies. The objective of this study was to evaluate the effect of three thermal profiles on development stages of this insect, allowing synchronization of cycle production. Two distinct rearing phases were done: firstly, using nucleous of incubation for development of eggs and, secondly, using circular-aluminum manifolds for development of larvae and pupae. The time necessary for development of the immature stages decreased with higher temperatures. Incubation periods lasted 13.4 days at 22 degrees C, 8.3 at 27 degrees C and 6.8 days at 32 degrees C, while periods for larvae development lasted 40.4, 27.2, and 23.4 days, respectively, for the same temperatures. Development to pupal stage was observed 18.2, 15.0, and 12.2 days, respectively, for the same temperatures. Larval survival was higher at 32 degrees C, however embryonic stages and pupae survival were higher at 27 degrees C. and 22 degrees C, respectively. The threshold temperature was 11.209167 degrees C for the embryonic development stage, 7.695869 degrees C for larval stage, and 1.943050 degrees C for pupal stage of G. mellonella. Thermal constants were 138.380533 DG (degree day) for egg, 554.968830 DG for larvae, and 369.054080 DG for pupae.