The life history of Labiostrongylus eugenii, a nematode parasite of the Kangaroo Island Wallaby (Macropus eugenii): The parasitic stages

Smales L. R. 1977. The life history of Labiostrongylus eugenii, a nematode parasite of the Kangaroo Island Wallaby (Macropus eugenii): the parasitic stages. International Journal for Parasitology7: 457–461. Labiostrongylus eugenii infective larvae ingested by the Kangaroo Island Wallaby exsheathe on the saccular stomach wall, then invade the mucosa causing one or two chronic irritative hyperpastic nodules, each containing a number of larvae. Six to eight weeks later the larvae leave the stomach wall and moult to fourth-stage. The fourth-stage larva is characterized by eight lips, while the oesophagus and anterior intestine assume the adult form. The posterior ends of late fourth-stage larvae become sexually differentiated. Moulting to adult occurs after 6–8 weeks with the first gravid females being observed 3 months later. The prepatent period appears to last about 6 months. Reasons for failure of experimental infection trials and the epidemiological significance of the life cycle were discussed.     

Parasitism of the mosquito Culex pipiens by the nematode Heterorhabditis bacteriophora

Various concentrations of the nematode Heterorhabditis bacteriophora were added to dishes containing second, third, and fourth larval instars of the mosquito, Culex pipiens, respectively. The infective stage nematodes were ingested by the mosquito larvae, they then penetrated through the alimentary tract in the neck region and entered the hemocoel. A melanization reaction killed many invading nematodes, but heavier concentrations overwhelmed the hosts' defense reaction and 100% mortality of third- and fourth-instar larvae was achieved using between 170 and 200 nematodes per host. Death was either due to the nematode releasing cells of the symbiotic bacterium, Xenorhabdus luminescens, into the hemocoel or to foreign bacteria (mostly Pseudomonas aeruginosa), which were introduced by the penetrating nematodes. The potential use of this nematode as a biological control agent of larval culicine mosquito is discussed.     

An ultrastructural study of the invasion of Ascaris suum larvae by neutrophils

Neutrophils are capable of penetrating the cuticle of the second-stage larvae of the nematode Ascaris suum. The integrity of the internal tissues of the larvae is then destroyed by cytoplasmic extensions of the neutrophils.     

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

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

Comparative ultrastructure of the oesophageal glands of third stage larval hookworms

Smith J. M. 1976. Comparative ultrastructure of the oesophageal glands of third stage larval hookworms. International Journal for Parasitology6: 9–13. The oesophageal glands of the third stage larvae of Necator americanus and Ancylostoma tubaeforme are compared, both before and after penetration through skin. The glands of “infective” larvae of N. americanus are densely packed with secretory granules, contrasting with a reduced gland size in the “penetrated” larvae coupled with the presence of gland secretions in the oesophageal lumen.No difference was observed between the glands of “infective” and “penetrated” larvae of A. tubaeforme. The role of oesophageal gland secretions for penetration of host skin is discussed.     

Effect of attachment of Corynebacterium rathayi on movement of Anguina agrostis larvae

Bird A. F. and Riddle D. L. 1984. Effect of attachment of Corynebacterium rathayi on movement of Anguina agrostis larvae. International Journal for Parasitology 14: 503–511. The movement of freshly hatched larvae (FHL₂) and dauer larvae (DL₂) of Anguina agrostis was compared on ‘agarose’ plates. The DL₂S moved faster and over greater distances. They were not attracted to Corynebacterium rathayi on agar plates, but contact with this bacterium, in most instances, markedly reduced their speed of movement. This reduction was found to be approximately proportional to the concentration of the bacteria (from 8 × 10⁵ to 8 × 10⁸ per ml) to which the DL₂ were exposed prior to observation of their movement, as was the number of bacteria observed to be adhering when viewed under the light microscope. This type of bacterial attachment appeared to be largely stage specific as it was much more pronounced and characteristically different in the DL₂ from that in the FHL₂. This interaction between the DL₂ and the bacterium was similar in material from fresh and from rehydrated nematode galls so that it was apparently not dependent on any surface cuticular changes associated with anhydrobiosis. Furthermore, pretreatment of the nematodes with the surfactant SDS did not influence attachment. An electron microscope study of sections cut through DL₂ exposed to bacteria showed that this interaction was indeed not a surface phenomenon but that the bacteria exerted a pathological effect on the nematode. The bacterium's capsular material actually penetrated and broke down the nematode epicuticle and part of the cortical zone. These observations make it easier to understand the dramatic physiological responses of this nematode to these bacteria.     

Observations on the occurrence of crystal-like structures in nematode parasites of sheep and cattle

Waller P. J., Dash K. M. and Major G. W. 1979. Observations on the Occurrence of crystal-like structures in nematode parasites of sheep and cattle. Internationl Journal for Parasitology9: 147–151. Intracellular rod-like inclusions were found in a high percentage of inhibited fourth-stage larvae of H. contortus in sheep with naturally acquired infections. Artificial infections showed inclusions occurred in developing as well as inhibited fourth-stage larvae. The presence of these structures was associated with degenerative changes of the parasites and serial worm counts showed that larvae with large numbers of inclusions failed to persist within the host.Large refractive hexagonal crystals were observed in the gut lumen of fourth-stage and adult O. ostertagi acquired by previously worm-free lambs grazed on cattle pastures. The crystals tended to accumulate in the posterior gut region where they may cause gut blockage and lead to early mortality of the parasites in the abnormal host. No inclusions or crystals were observed in O. ostertagi from cattle or goats, or in O. circumcincta from sheep.     

Susceptibility of early larval stages of Pseudaletia unipuncta and Spodoptera exigua (Lepidoptera: Noctuidae) to the entomogenous nematode Steinernema feltiae (Rhabditida: Steinernematidae)

Early stages (neonate to 7- or 8-day-old larvae) of Spodoptera exigua and Pseudaletia unipuncta were exposed to the entomogenous nematode, Steinernema feltiae, at concentrations of 0, 10, 25, 60, 100, or 200 nematodes per larva. Larvae of both species were susceptible to nematode infections. However, neonate larvae of S. exigua were significantly less susceptible to nematode infection than 3- or 8-day-old larvae at or above 50 nematodes per larva. Mortalities of neonate larvae exposed to 50 or more nematodes ranged from 68 to 74% while mortalities of 3- and 8-day-old larvae ranged from 91 to 100%. The results with P. unipuncta showed similar trends as described for S. exigua, albeit at a lower mortality level and usually with no statistical differences. Mortalities of neonate larvae exposed to 50 or more nematodes ranged from 34 to 44% while mortalities of 7-day-old larvae ranged from 32 to 91%.     

The reproduction potentials of four entomopathogenic nematode strains related to cost-effective production for biological control

Bioassays to evaluate the mortality, virulence and reproduction potentials of four indigenous EPN strains, S-PQ16, S-BM12, H-KT3987 and H-CB3452 on insect larvae of mealworm (Tenebrio molitor) and greater wax moth (Galleria mellonella) revealed the highest mortality rates of two insect larvae at the highest inoculation dose of 100 IJs to range from 89 to 100 percent and 94.3–100 percent at 48 h after inoculation, respectively. Virulence was high for all nematode strains, with LC₅₀ values between 29.6 and 47.3 IJs/insect host. The highest IJ yields were different between nematode strains and insect host, from 66.8 × 10³ IJs (S-PQ16) to 118.6 × 10³ IJs (H-KT3987) on T. molitor, and from 54.2 × 10³ IJs (S-BM12) to 163.3 × 10³ IJs (H-KT3987) on G. mellonella. The culturing cost in terms of food expenditure for rearing insect larvae varied between insect larvae and nematode strains, from 6.76 to 26.63 USD per billion IJs for nematode strains cultured on T. molitor larvae and from 3.54 to 7.81 USD per billion IJs for nematode strains cultured on G. mellonella larvae. The full cost for a nematode product of 2.5 × 10⁹ IJs per hectare, produced through in vivo mass culturing, of the most efficient nematode strain, H-KT3987, was 191.3 USD, slightly cheaper than 199.4 USD for the same nematode product produced through in vitro mass culturing.     

Subclinical nematode parasitism affects activity and rumination patterns in first-season grazing cattle

Sickness behaviour has been suggested as an applicable indicator for monitoring disease. Deviating feeding behaviour and activity can provide information about animals’ health and welfare status. Recent advances in sensor technology enable monitoring of such behaviours and could potentially be utilized as an indicator of gastrointestinal nematode (GIN) infections. This study investigated activity and rumination patterns in first-season grazing steers exposed to subclinical infection levels of the GIN Ostertagia ostertagi and Cooperia oncophora. At turnout, animals were allocated to one of four experimental groups and were faced with “high” (H1, n = 15; H2, n = 17) or “low” (L1, n = 17; L2, n = 11) levels of parasite exposure by grazing in similar enclosures contaminated with overwintering third stage (L3) GIN larvae. Animals in H1 and H2 (HP) received a 1:1 mix of approximately 10,000 O. ostertagi and C. oncophora L3 at turnout; whereas the animals in L1 and L2 (LP) were treated monthly with ivermectin. Activity and rumination patterns were monitored by fitting animals with leg- (IceQube) and neck-mounted (Heatime) sensors. BW was recorded every fortnight, whereas faecal and blood samples were examined every four weeks for nematode faecal egg count and serum pepsinogen concentrations (SPCs). There was an interaction effect of exposure level and period (P < 0.0001) on average lying daily time across the entire grazing time. A higher mean daily lying time (P = 0.0037) was found in HP compared with LP during the first 40 days on pasture. There was also interaction effects of treatment and day since turnout on rumination time (P < 0.0001) and rumination change (P = 0.0008). Also mean daily steps (P < 0.0001) and mean daily motion index (P < 0.0001) were markedly higher in HP during days 62–69, coinciding with peaking SPC in HP. Strongyle eggs were observed both in HP and LP from 31 days after turnout. Eggs per gram (EPG) differed between parasite exposure levels (P < 0.0001), with mean EPG remaining low in LP throughout the experiment. Similarly, an increase in SPC was observed (P < 0.0001), but only in HP where it peaked at day 56. In contrast, no difference in BW gain (BWG) (P = 0.78) between HP and LP was observed. In conclusion, this study shows that behavioural measurements monitored with sensors were affected even at low infection levels not affecting BWG. These combined results demonstrate the potential of automated behavioural recordings as a tool for detection of subclinical parasitism.     

Biological Control of the Leafminer Liriomyza trifolii (Burgess): Implications for Intraguild Predation between Diglyphus begini Ashmead and Steinernema carpocapsae (Weiser)

Studies were conducted on the combined use of the eulophid parasitoid wasp Diglyphus begini Ashmead and the entomopathogenic nematode Steinernema carpocapsae (Weiser) for control of the leafminer Liriomyza trifolii (Burgess) on chrysanthemums. Several factors indicated that these two agents were suitable for combined use: adult D. begini were not susceptible to nematode infection, leafminer larvae parasitized by the wasp were less susceptible to nematode infection, adult wasps detected and tended to avoid ovipositing on nematode-infected leafminer larvae, nematode-infected larvae served as host-feeding sources for the adult wasps, and nematodes showed equal orientation toward paralyzed/parasitized leafminer larvae and healthy leafminer larvae. However, interspecific interference and intraguild predation (IGP) between the agents were found. Infection of D. begini larval stages by nematodes was seen in petri dishes and in intact leaf mines. The presence of nematodes in mines with wasp eggs decreased the chance of wasp survival to adulthood. IGP may be minimized through proper timing of natural enemy releases.     

Effect of repeatedly sensitized bursal extract on the resistance of WLH chickens to experimental infection of Ancylostoma caninum larvae

Goyal P. K. and Johri G. N. 1982. Effect of repeatedly sensitized bursal extract on the resistance of WLH chickens to experimental infection of Ancylostoma caninum larvae. International Journal for Parasitology12: 245–249. Bursal extract from chickens, repeatedly sensitized with Ancylostoma caninum larvae and from normal nonsensitized donor WLH chickens were injected separately into experimental and control groups of isologous recipients of same age and weight. Extract from donors infected with repeatedly high dose (250 + 250 + 500) and low dose (125 + 125 + 250) of A. caninum larvae produced significantly greater acquired immunity in recipients compared to those which received nonsensitized (normal) extract. However, no significant difference was observed between the two experimental groups.     

Effects of Inoculum Level and Time of Microcoleus vaginatus on Control of Meloidogyne incognita on Tomato

Application of Microcoleus vaginatus, a blue-green alga (Cyanobacterium) at different levels along with Meloidogyne incognita, second stage larvae, in the rhizosphere of tomato plants; showed that the plant growth as well as yield of tomato were increased and gall formations and nematode populations decreased with the increase in inoculum level of M. vaginatus. An inoculum level of 20 ml endospores suspension of M. vaginatus (2.4 × 10⁶ endospores per ml) per plant was optimum to reduce nematode attack with a population density of 1000 larvae per kg soil. Plant growth and yield of fruits were greatly suppressed and gall formations on roots, and nematode populations in soil were increased when M. incognita larvae added five days prior to M. vaginatus inoculation. On the other hand, when M. vaginatus inoculated ten days before nematode inoculation, suppressive effect of M. incognta on plants was reduced and their population density as well as gall formations were also decreased significantly. The efficacy of simultaneous inoculation of both nematode and M. vaginatus was lied in between two treatments discussed above.     

Nematode parasites from the skin of the platypus Ornithorhynchus anatinus (Monotremata)

The morphology of three nematode taxa found in the skin of the platypus Ornithorhynchus anatinus (Monotremata) in southeastern New South Wales, Australia, is described and illustrated. Fourth-stage larvae belonging to the Rhabditoidea, ?Cylindrocorporidae, occurred primarily in superficial keratin and epidermal layers of the skin and in hair follicles. A nematode recognised by its conspicuous synlophe as a fourth-stage larva of the Trichostrongyloidea, of unknown genus and species but probably belonging to the molineid evolutionary line, was observed in histological section in the hypodermis. A gravid female nematode belonging to the Filarioidea, ?Lemdaninae, occurred in a lymphoid nodule in the hypodermis. O. anatinus is a protected species and these data are presented to draw the attention of parasitologists to skin as an important habitat for nematodes of the platypus.     

Vaccination against the nematode Trichostrongylus colubriformis. III. Some observations on factors influencing immunity to infection in vaccinated guinea-pigs

Rothwell T. L. W. 1978. Vaccination against the nematode Trichostrongylus colubriformis. III. Some observations on factors influencing immunity to infection in vaccinated guineapigs. International Journal for Parasitology8: 33–37. Guinea-pigs were protected against infection with T. colubriformis when soluble material extracted from fourth-stage larvae was administered by the subcutaneous, intradermal, intraperitoneal and intraduodenal but not oral routes. The level of immunity following vaccination by the various effective routes was similar. Mature animals were found to respond significantly better to vaccination than immature animals. Significant immunity was present 10 days after vaccination but higher levels were found after 20 and 40 days. A single dose of vaccine was as effective as three divided doses. Finally, it was found that the adjuvant aluminium hydroxide gel, but not B. pertussis vaccine or levamisole improved the level of immunity to infection which followed vaccination.     

Kinetics of primary and secondary infections with Strongyloides ratti in mice

Dawkins H. J. S. and Grove D. I. 1981 Kinetics of primary and secondary infections with Strongyloides ratti in mice. International journal for Parasitology11: 89–96. The kinetics of infection with S. ratti were quantitated in normal and previously exposed C57B1 /6 mice. In primary infections, larvae penetrated the skin rapidly and were seen in peak numbers 12 h after infection. By 24 h after infection, larval numbers had declined appreciably and there was a slow decrease in numbers thereafter. Larvae were first observed in the lungs at 24 h and maximal recovery occurred at 48 h. It is thought that larval migration through the lungs is rapid. Worms were first seen in the intestines two days after infection. Maximum numbers were seen on the fifth day and worm expulsion was complete by day 10. Two moults took place in the small intestine during days 3 and 4 after infection. Rhabditiform larvae were first noted on the fourth day after infection. Mice exposed to S. ratti four weeks previously had significantly less larvae in the skin 4 and 12 h after infection but by 24 h there was no difference when compared with mice with primary infections. Peak recovery of larvae from the lungs occurred 24 h after infection; significantly less larvae were recovered on days 2 and 3 when compared with normal mice. There was a marked reduction in the adult worm burden in the gut; the number of worms recovered was less than one fifth of that seen in primary infections. Those worms which did mature were less fecund and were expelled from the intestines within 7 days of infection. It is suggested that in previously exposed animals, the migration of larvae from the skin is hastened, many of these larvae are destroyed in the lungs and that expulsion of worms which do mature in the intestines is accelerated.     

Heterorhabditis sp. (Nematoda: Heterorhabditidae): A Nematode Parasite Isolated from the Banded Cucumber Beetle Diabrotica balteata

A nematode identified as Heterorhabditis sp. was discovered in June 1982 in larval cadavers of the banded cucumber beetle, Diabrotica balteata, in soil on wooded land. Effective beetle control (over 95%) was obtained when larvae were exposed to potted soil containing infective stage nematode juveniles or infected larval cadavers. The nematode was propagated in vivo on larvae of D. balteata, Diaphania nitidalis (the pickleworm), and Galleria mellonella (the greater wax moth). This Heterorhabditis sp. has promising potential as a biocontrol agent for the banded cucumber beetle.     

Histopathological appearances in primary and secondary infections with Strongyloides ratti in mice

Dawkins H. J. S., Muir G. M. & Grove D. I. 1981. Histopathological appearances in primary and secondary infections with Strongyloides ratti in mice. International Journal for Parasitology11: 97–103. The histological appearances of the skin, lungs and small intestines of mice with primary and secondary infections with S. ratti are described. When the skins of mice with a primary infection were examined, larvae were seen scattered throughout the dermis. An inflammatory reaction of neutrophils and eosinophils was first noted around larvae 12 h after infection. By 48 h, mononuclear cells were prominent. The intensity of the inflammatory reaction gradually increased to a maximum on the fifth day and the larvae were destroyed. Very few larvae were seen in the lungs; those observed were located in the alveolar spaces and were not surrounded by an inflammatory infiltrate. Worms in the small intestines were found mostly in the crypts of Leiberkuhn, and were probably located within the epithelial layer; there was no significant villous atrophy or cellular infiltration. Marked differences were found in the tissues of mice with secondary infections. In the skin, oedema and neutrophils and eosinophils were seen around worms as early as 2 h after infection. By 24 h after infection, there was a mixed inflammatory infiltrate and worms were undergoing disintegration. Larvae in the lungs were surrounded by polymorphonuclear and mononuclear cells 48 h and 72 h after infection and the engulfed larvae were undergoing lysis. Only a few worms were seen in the intestines of mice with a secondary infection; the histological appearances were similar to that found in animals with primary infections. It is suggested that the rapid development of an oedematous reaction in the skins of immune mice may facilitate the entry of larvae into the bloodstream and that inflammatory cells destroy many larvae in the lungs of immune mice.     

Susceptibility of the Plum Curculio,Conotrachelus nenuphar,to Entomopathogenic Nematodes

The plum curculio, Conotrachelus nenuphar, is a major pest of pome and stone fruit. Our objective was to determine virulence and reproductive potential of six commercially available nematode species in C. nenuphar larvae and adults. Nematodes tested were Heterorhabditis bacteriophora (Hb strain), H. marelatus (Point Reyes strains), H. megidis (UK211 strain), Steinernema riobrave (355 strain), S. carpocapsae (All strain), and S. feltiae (SN strain). Survival of C. nenuphar larvae treated with S. feltiae and S. riobrave, and survival of adults treated with S. carpocapsae and S. riobrave, was reduced relative to non-treated insects. Other nematode treatments were not different from the control. Conotrachelus nenuphar larvae were more susceptible to S. feltiae infection than were adults, but for other nematode species there was no significant insect-stage effect. Reproduction in C. nenuphar was greatest for H. marelatus, which produced approximately 10,000 nematodes in larvae and 5,500 in adults. Other nematodes produced approximately 1,000 to 3,700 infective juveniles per C. nenuphar with no significant differences among nematode species or insect stages. We conclude that S. carpocapsae or S. riobrave appears to have the most potential for controlling adults, whereas S. feltiae or S. riobrave appears to have the most potential for larval control.     

Parasitism of Western Corn Rootworm Larvae and Pupae by Steinernema carpocapsae

Virulence and development of the insect-parasitic nematode, Steinernema carpocapsae (Weiser) (Mexican strain), were evaluated for the immature stages of the western corn rootworm, Diabrotica virgifera virgifera LeConte. Third instar rootworm larvae were five times more susceptible to nematode infection than second instar larvae and 75 times more susceptible than first instar larvae and pupae, based on laboratory bioassays. Rootworm eggs were not susceptible. Nematode development was observed in all susceptible rootworm stages, but a complete life cycle was observed only in second and third instar larvae and pupae. Nematode size was affected by rootworm stage; the smallest infective-stage nematodes were recovered from second instar rootworm larvae. Results of this study suggest that S. carpocapsae should be applied when second and third instar rootworm larvae are predominant in the field.