Trichinella papuae n.sp. (Nematoda), a new non-encapsulated species from domestic and sylvatic swine of Papua New Guinea

Encapsulated and non-encapsulated species of the genus Trichinella are widespread in sylvatic animals in almost all zoogeographical regions. In sylvatic animals from Tasmania (Australian region), only the non-encapsulated species Trichinella pseudospiralis has been reported. Between 1988 and 1998, non-encapsulated larvae of Trichinella were detected in five domestic pigs and six wild boars from a remote area of Papua New Guinea. Morphological, biological, and molecular studies carried out on one strain isolated from a wild boar in 1997 suggest that these parasites belong to a new species, which has been named Trichinella papuae n.sp. This species can be identified by the morphology of muscle larvae, which lack a nurse cell in host muscles, and whose total length is one-third greater than that of the other non-encapsulated species, T. pseudospiralis. Adults of T. papuae do not cross with adults of the other species and genotypes. Muscle larvae of T. papuae are unable to infect birds, whereas those of T. pseudospiralis do. The expansion segment V of the large subunit of the ribosomal DNA differs from that of the other species and genotypes. All of these features allow for the easy identification of T. papuae, even in poorly equipped laboratories. The discovery and identification of a second non-encapsulated species in the Australian region strongly supports the existence of two evolutionary lines in the genus Trichinella, which differ in terms of the capacity of larvae to induce a modification of the muscle cell into a nurse cell.     

Stand tall and they still get you in your Achilles foot-pad.

The free-living first-instar larvae of Strepsiptera (Insecta) are the infective stage of the parasitoid. They normally enter the host via the abdominal cuticle, and there have also been reports of entry via the egg of the host. The first-instar larvae of Stichotrema dallatorreanum Hofeneder in Papua New Guinea were found to enter the host orthopteran via the tarsi. This is, to my knowledge, the first report of entry of first-instar larvae of Strepsiptera via the attachment pads (euplantulae) of the host.     

Biological variation in Trichinella species and genotypes

At present, the genus Trichinella comprises seven species of which five have encapsulated muscle larvae (T. spiralis, T. nativa, T. britovi, T. nelsoni and T. murrelli) and two do not (T. pseudospiralis and T. papuae) plus three genotypes of non-specific status (T6, T8 and T9). The diagnostic characteristics of these species are based on biological, biochemical and genetic criteria. Of biological significance is variation observed among species and isolates in parameters such as infectivity and immunogenicity. Infectivity of Trichinella species or isolates is determined, among other considerations, by the immune status of the host in response to species- or isolate-specific antigens. Common and particular antigens determine the extent of protective responses against homologous or heterologous challenge. The kinetics of isotype, cytokine and inflammatory responses against T. spiralis infections are isolate-dependent. Trichinella spiralis and T. pseudospiralis induce different dose-dependent T-cell polarizations in the early host response, with T. spiralis initially preferentially promoting Th1-type responses before switching to Th2 and T. pseudospiralis driving Th2-type responses from the outset.     

Alterations in the longevity and fecundity of adult Trichinella pseudospiralis related to method of isolation of infective larvae

The infectivity of Trichinella pseudospiralis infective larvae was reduced significantly following exposure to low pH or a combination of 1% pepsin at low pH compared to that for larvae isolated in phosphate-buffered saline (PBS) at pH 7.0. Reduction of host gastric pH by administration to mice of sodium bicarbonate solution in PBS was accompanied by an increase in the infectivity of larvae isolated in 1% pepsin/HCl (P/HCl) compared to that for worms inoculated into hosts given PBS alone. Fewer adult worms developing from larvae isolated in P/HCl became established in the host small bowel than was seen with larvae isolated in PBS; moreover, the fecundity in vitro of adult worms developing from P/HCl-isolated larvae was reduced below that for adults developing from larvae isolated from host muscle in PBS. More adult worms were recovered following infection of immune hosts with PBS-isolated larvae than were recovered from immune mice challenged with larvae isolated in P/HCl. Similar findings were observed in mice immunized by infection with Trichinella spiralis and challenged with T. pseudospiralis larvae isolated in either P/HCl or PBS. Immunization of mice with T. pseudospiralis larvae isolated by either method and challenged with larvae of T. spiralis resulted in recovery of similar percentages of the challenge inoculum.     

Trichinella spiralis: effect of high temperature on infectivity in pork

Twenty gram samples of homogenized Boston shoulder from swine experimentally infected with Trichinella spiralis were sealed in plastic pouches, pressed to a uniform thickness of 2 mm, and subjected to water bath temperatures of 49, 52, 55, 60, and 63 +/- 0.5 C for intervals of 2 min to 6 hr, especially within the interval of 0 to 15 min. These times included a period of about 1 min at the start and a period of about 1 min at the end for temperature equilibration. Treated samples were rapidly chilled to 25 C and then digested in a 1% pepsin-HCl solution at 37 C for 18 hr to recover T. spiralis larvae. The recovered larvae were suspended in 2 ml saline; 1 ml of this suspension was introduced into the stomach of each of two rats. The linear equation, log (time) = 17.3 -0.302 (temperature), was calculated from the time required at each temperature for the inactivation of T. spiralis larvae. The correlation coefficient for that relationship was r = -0.994. Larvae heated in the meat to 55 C for 4 min retained their infectivity, but were rendered noninfective after 6 min at 55 C. At 60 C, larvae were not infective after only 2 min (zero dwell time); whereas at 52 C, 47 min were required to render the larvae noninfective. Larvae in meat heated to 49 C were infective after 5 hr but not after 6 hr. These data demonstrate that the destruction of infectivity of T. spiralis is time-temperature related.     

Tolerance to low temperatures of domestic and sylvatic Trichinella spp. in rat muscle tissue

The present study was designed to investigate the tolerance to low temperatures of 9 Trichinella isolates in rat muscle tissue. Nine groups of 24 rats were infected with encapsulated Trichinella spiralis, Trichinella nativa, Trichinella britovi, Trichinella murrelli, Trichinella T6, Trichinella nelsoni, and 3 nonencapsulated Trichinella pseudospiralis strains. Six rats from each of the groups were necropsied at 5, 10, 20, and 40 wk postinfection (wpi). Muscle tissues containing Trichinella larvae were exposed to temperatures of -18, -5, and 5 C for 1 or 4 wk, and afterward the reproductive capacity index (RCI) in mice was determined for the 9 individual Trichinella isolates. Only T. nativa muscle larvae were infective after freezing at a temperature of -18 C. At 5 wpi all encapsulated isolates, except for the tropical species T. nelsoni, remained infective after exposure to a temperature of -5 C for both 1 and 4 wk, whereas nonencapsulated T. pseudospiralis survived only 1 wk of exposure. All Trichinella spp. remained infective after exposure to a temperature of 5 C. Muscle larvae for all investigated species remained infective as long as they persisted in live rats during the experiment. Analysis of variance showed a significant effect of age on the temperature tolerance of encapsulated T. spiralis and nonencapsulated T. pseudospiralis. In addition, significant interaction between age of muscle larvae and length of exposure was found. In general Trichinella muscle larvae of medium age (10 and 20 wpi) tolerated freezing better than early and late stages of infection (5 and 40 wpi). This is the first study to demonstrate such a relationship between age of infection and temperature tolerance of Trichinella spp. muscle larvae.     

Trichinella britovi etiological agent of sylvatic trichinellosis in the Republic of Guinea (West Africa) and a re-evaluation of geographical distribution for encapsulated species in Africa

In West Africa, Trichinella infection was documented in humans and animals from Senegal in the 1960s, and the biological characters of one isolate showed a lower infectivity to domestic pigs and rodents when compared with that of a Trichinella spiralis pig isolate from Europe. To identify the Trichinella species present in West Africa, a survey was conducted in a total of 160 wild animals in the Republic of Guinea. Three Viverridae, one true civet (Viverra civetta) and two African palm civets (Nandinia binotata) from the Fouta Djallon Massif, Pilimini Subprefecture, were found positive by artificial digestion of muscle samples. Trichinella larvae from these three viverrids were identified as Trichinella britovi and no difference was detected in three examined sequences from these African isolates and the reference strain of T. britovi from Europe, indicating common ancestry, an historically continuous geographic distribution, and recent isolation for African and European populations. The detection of T. britovi in West Africa modifies our knowledge about the distribution of encapsulated species of Trichinella in Africa. Thus, Trichinella nelsoni is now considered to have a distribution limited to the Eastern part of the Afrotropical region from Kenya to South Africa. This provides a plausible explanation for the presence of Trichinella T8 in Namibia and South Africa, and further suggests that T. britovi could be the Trichinella species circulating among wild animals of Northern Africa.     

The endemic normal in lymphatic filariasis: A static concept

Residents of areas endemic for lymphatic filariasis are continually exposed to infection with mosquito-transmitted infective larvae (L3), some of which survive to become adult worms and subsequently produce micro filarial (mf) transmission stages. The question of whether naturally acquired resistance occurs in adult residents of endemic areas has recently become of interest as the development of molecular vaccines against filarial parasites is being considered(1,2). There have been two epidemiological approaches to demonstrate acquired resistance to Filariasis in human populations. In this review Karen Day examines both approaches in the context of an immunoepidemiological study of bancroftian filariasis in Papua New Guinea (PNG). The merits of each as a conceptual framework for studies of protective immunity in lymphatic filariasis will be discussed.     

Nonenzymatic isolation of Trichinella pseudospiralis infective L1 larvae at pH 7.4

Over half of the number of Trichinella pseudospiralis infective L1 larvae recovered from host carcasses by pepsin-HCl digestion were isolated from homogenized carcasses incubated in HBSS. More worms isolated by the latter method were viable compared to those isolated by pepsin-HCl digestion. When host carcasses infected with T. pseudospiralis were diced into pieces and incubated in HBSS, 30% more worms were recovered than from homogenized carcasses incubated in HBSS as above, and the majority of worms acquired by the former method were viable. The infectivity of T. pseudospiralis infective L1 larvae isolated from homogenized muscle in HBSS was 3.9 times greater than that for larvae recovered from homogenized carcasses by pepsin-HCl digestion. Only 4% and 0.8% of the number of T. spiralis recovered from homogenized muscle by pepsin-HCl digestion were isolated from homogenized or diced muscle incubated in HBSS, respectively. Fewer T. spiralis isolated from homogenized tissue in HBSS were viable compared to those recovered from homogenized carcasses digested in pepsin-HCl or diced carcasses incubated in HBSS.     

Parasitic zoonoses in Papua New Guinea

Relatively few species of zoonotic parasites have been recorded in humans in Papua New Guinea. A greater number of potentially zoonotic species, mostly nematodes, occur in animals but are yet to be reported from humans. Protozoa is the best represented group of those infecting man, with Giardia duodenalis, Cryptosporidium parvum, Cyclospora cayetanesis, Toxoplasma gondii, Sarcocystis spp., Entamoeba polecki, Balantidium coli and, possibly, Blastocystis hominis. The only zoonotic helminths infecting humans include the trematode Paragonimus westermani, the cestodes Hymenolepis nana, H. diminuta and the sparganum larva of Spirometra erinacea, and the nematodes Trichinella papuae and Angiostrongylus cantonensis and, possibly, Ascaris suum. Other groups represented are Acanthocephala (Macracanthorhynchus hirudinaceus)), insects (Chrysomya bezziana, Cimex sp., Ctenocephalides spp.), and mites (Leptotrombidium spp. and, possibly Sarcoptes scabiei, and Demodex sp.). One leech (Phytobdella lineata) may also be considered as being zoonotic. The paucity of zoonotic parasite species can be attributed to long historical isolation of the island of New Guinea and its people, and the absence until recent times of large placental mammals other than pig and dog. Some zoonotic helminths have entered the country with recent importation of domestic animals, in spite of quarantine regulations, and a few more (two cestodes, one nematode and one tick) are poised to enter from neighbouring countries, given the opportunity. Improvement in water supplies, human hygiene and sanitation would reduce the prevalence of many of these parasites, and thorough cooking of meat would lessen the risk of infection by some others.     

Sylvatic and domestic Trichinella spp. in wild boars; infectivity, muscle larvae distribution, and antibody response

Thirty-six wild boars were inoculated with Trichinella spiralis, Trichinella nativa, Trichinella britovi, Trichinella pseudospiralis (USSR), T. pseudospiralis (USA), T. pseudospiralis (AUST), Trichinella murrelli, Trichinella T6, and Trichinella nelsoni. The wild boars were killed at 5 and 10 wk postinoculation (PI), and the number of muscle larvae per g (lpg) of tissue was determined for 18 muscles or muscle groups. Five weeks PI, all Trichinella genotypes had established as muscle larvae, but their infectivity varied widely: T. spiralis established in high numbers (mean = 296 lpg), T. britovi, T. nelsoni, and 1 of the T. pseudospiralis genotypes (AUST) in moderate numbers (mean = 53-74 lpg), whereas the remaining genotypes were poorly infective (mean 2-16 lpg). Because of considerable weight gain of the wild boars, an estimated total larval burden (live weight x lpg) was calculated for each animal. The total larval burden did not change significantly over time for T. spiralis, T. murrelli, T. britovi, T. nelsoni, and T. pseudospiralis (USA and USSR), whereas a significant reduction could be demonstrated for T. nativa, Trichinella T6, and T. pseudospiralis (AUST). Diaphragm and tongue were predilection sites in wild boars, independent of Trichinella genotype and infection level. At low infection levels, a greater percentage of larvae were found in diaphragm and tongue at 10 wk than 5 wk PI. Antibody responses increased rapidly between weeks 3 and 5 PI. For T. spiralis and T. nelsoni, the high antibody level persisted throughout the experimental period, but for T. nativa, T. britovi, T. murrelli, or Trichinella T6, the levels declined. For T. pseudospiralis, the antibody response increased more gradually between weeks 3 to 10 PI. Infection with all genotypes of Trichinella were detected using any of 7 excretory-secretory antigens, which points to the potential use of 1 common antigen for epidemiological studies on Trichinella in wild boars. In conclusion, T. spiralis is highly infective to wild boars, T. britovi, T. nelsoni, T. pseudospiralis (USA), and T. pseudospiralis (USSR) are moderately infective, and T. nativa, T. murrelli, T. pseudospiralis (AUST), and Trichinella T6 are poorly adapted to this host species.     

Trichinella pseudospiralis populations of the Palearctic region and their relationship with populations of the Nearctic and Australian regions

Since few non-encapsulated isolates of Trichinella have been studied to date, their level of differentiation from encapsulated species and the taxonomic value of the observed polymorphisms remain to be determined. To this end, biological, biochemical and molecular data from 11 isolates of Trichinella pseudospiralis and one isolate of Trichinella papuae were examined using the broad group of encapsulated species and genotypes for comparison. Single-worm cross-breeding experiments and reproductivity capacity indices revealed F1 progeny only among T. pseudospiralis isolates from different zoogeographical regions, whereas no F1 were produced when T. pseudospiralis was crossed with T. papuae. Furthermore, unlike T. pseudospiralis, T. papuae failed to infect chickens. Comparative analysis of 12 allozymes revealed a single difference between Nearctic and Australian isolates of T. pseudospiralis, but substantial differences when compared with T. papuae (i.e. two unique and six diagnostic markers). Molecular studies involving mitochondrial-derived genes encoding cytochrome oxidase I and the large subunit ribosomal DNA indicated a high level of sequence similarity among T. pseudospiralis isolates; however, a concomitantly high level of variation was observed in expansion segment five of the genomic large subunit ribosomal DNAs among T. pseudospiralis isolates and between this species and T. papuae. Collectively, these results demonstrate high uniformity among isolates of T. pseudospiralis from Eurasia and polymorphism among isolates of T. pseudospiralis belonging to different zoogeographical regions; the results corroborate the classification of T. papuae as a differentiated species.     

Heat shock protein synthesis over time in infective Trichinella spiralis larvae raised in suboptimal culture conditions

Changes in the viability, infectivity and heat shock protein (Hsp) levels are reported in Trichinella spiralis first stage larvae (L1) stored in 199 medium for up to seven days at 37 degrees C. These conditions induce stress that the larvae, eventually, cannot overcome. After three days of storage, the infectivity and viability were unchanged, although higher Hsp70 levels were observed. After this time, larvae gradually lost viability and infectivity, coinciding with a decrease in Hsp70 and Hsp90 and an increase in actin (a housekeeping protein). In addition, a possibly inducible heat shock protein, Hsp90i, appeared as constitutive Hsp90 disappeared. No significant changes in Hsp60 levels were detected at any time. These results suggest that heat shock proteins initially try to maintain homeostasis, but on failing, may be involved in cell death.     

Influence of temperature on the survival and infectivity of Trichinella spiralis larvae in Sarcophaga argyrostoma (Diptera, Sarcophagidae) maggots

To investigate the role of fleshfly maggots as a paratenic host for Trichinella spiralis larvae, maggots of Sarcophaga argyrostoma (Muscidae, Sarcophagidae) kept at different temperatures (26, 22, 20, 16, 12, 8, and 4 C) were allowed to feed on T. spiralis-infected mouse meat. Trichinella larvae found in maggots kept at 8-26 C were able to cause infection when inoculated in mice. Infective larvae survived in maggots up to 5 days postinfection at 8 C and for shorter periods of time at higher temperatures. The survival time in maggots was negatively related to the temperature of maggot breeding. The results suggest that the role of S. argyrostoma in the dissemination of Trichinella larvae in nature is limited in comparison to the role played by mammals with scavenger and cannibalistic behavior.     

Infectivity, persistence, and antibody response to domestic and sylvatic Trichinella spp. in experimentally infected pigs

Groups of pigs were inoculated with genotypes of Trichinella belonging to: Trichinella spiralis, Trichinella nativa, Trichinella britovi, Trichinella pseudospiralis (from Caucasus), T. pseudospiralis (from USA), Trichinella murrelli, Trichinella sp. (from North America), and Trichinella nelsoni. The pigs were sacrificed between 5 and 40weeks p.i., and the number of muscle larvae per gram (l.p.g.) of tissue was determined as an average of 18 muscles. All Trichinella genotypes were infective for pigs, but both their infectivity and persistence varied: 5weeks p.i., T. spiralis muscle larvae were present in high numbers (mean=427l.p.g.), while T. britovi, T. nelsoni, and T. pseudospiralis larvae were present in moderate numbers (means=24-52l.p.g.); larvae of the remaining genotypes were recovered only in low numbers (means=0.05-5. 00l.p.g.). The total larval burden (live weight of pigxl.p.g.) was constant over time for T. spiralis, T. britovi, and T. nelsoni, but declined significantly (P<0.05) for the other genotypes. Antibody responses could be detected 3-4weeks p.i. by seven different Trichinella ES antigens, but the antibody levels and dynamics differed significantly among the experimental groups. In pigs inoculated with T. spiralis, T. britovi, or T. nelsoni, the antibody level increased rapidly between weeks 3 and 5 p.i. and was stable or increased slightly throughout the experimental period. In pigs inoculated with T. nativa, T. murrelli, or Trichinella (T6) (from North America), a rapid increase was detected between weeks 3 and 5 p.i., but for these genotypes a reduction in the antibody levels was seen thereafter. In the pigs inoculated with T. pseudospiralis, the antibody level increased more gradually over a period from week 3 p. i. to weeks 15-20 p.i., and decreased thereafter. In general, all species of Trichinella were detected by any of the seven ES antigens, which points to the potential use of one common antigen for surveillance and epidemiological studies on both domestic and sylvatic Trichinella in pigs. Homologous ES antigens were slightly more sensitive in detecting antibodies to the corresponding Trichinella species.     

Characterization of a noncyst-forming isolate of Trichinella from a wild boar in Yugoslavia.

An isolate of Trichinella obtained from a wild boar in Yugoslavia did not form cysts in the musculature of its natural host. Subsequent inoculation into experimental hosts demonstrated that some larvae became encysted only after extended time periods, whereas others remained unencapsulated. Histological staining of larvae in the musculature demonstrated no deposition of collagen typically seen for Trichinella spiralis spiralis, Trichinella spiralis nativa, or Trichinella spiralis nelsoni. The Yugoslavian isolate, given the name of Zagreb isolate after the University where it was first studied, had low infectivity for pigs and mice. Isozyme analysis demonstrated greater homology with T. s. nelsoni than with other subspecies of Trichinella. Restriction fragment length polymorphisms and dot blot analyses further demonstrated the distinctive nature of this isolate. These results suggest that lack of cyst formation might be characteristic of isolates other than those designated Trichinella pseudospiralis and that this character might be important in the classification of Trichinella.     

A possible role for rusa deer (Cervus timorensis russa) and wild pigs in spread of Trypanosoma evansi from Indonesia to Papua New Guinea

Movement of transmigrants and livestock from western Indonesia to southeastern areas of Irian Jaya near the border with Papua New Guinea may pose a risk of introducing Trypanosoma evansi into Papua New Guinea via feral Rusa deer (Cervus timorensis russa) and wild pigs which inhabit these areas in large numbers. Pilot experimental studies were conducted to observe infection in pigs and Rusa deer with a strain of T. evansi isolated in Indonesia. Parasitaemia and signs of clinical disease were monitored each second day for 120 days. Trypanosomes were observed in haematocrit tubes at the plasma-buffy coat interface of jugular blood of deer and pigs on 86% and 37% of sampling occasions respectively. Parasitaemia was at a high level in deer for 35% of the time but for only 11.5% of the time in pigs. Results indicate that both Rusa deer and pigs have a high tolerance for infection with T. evansi. The deer suffered mild anaemia evidenced by a 25% reduction in packed cell volume (PCV) 14 days after infection which coincided with the initial peak in parasitaemia. However, PCV had returned to pre infection values by the end of the experiment. The pigs showed no change in PCV. There were no visual indications of disease in either species and appetite was not noticeably affected. It was concluded that both Rusa deer and pigs were capable reservoir hosts for T. evansi but that Rusa deer, with their more persistent higher levels of parasitaemia, have more potential to spread T. evansi into Papua New Guinea from West Irian than pigs.     

Observations on the morphology and infectivity for cattle of Babesia bovis parasites in unfed Boophilus microplus larvae after incubation at various temperatures

Dalgliesh R. J. and Stewart N. P. 1979. Observations on the morphology and infectivity for cattle of Babesia bovis parasites in unfed Boophilus microplus larvae after incubation at various temperatures. International Journal for Parasitology9: 115–120. The temperature of incubation of unfed Boophilus microplus larvae infected with Babesia bovis influenced the morphology and infectivity of the Babesia within the tick. Incubation at 37°C for 1–3 days stimulated the development of parasites morphologically similar to those usually observed in fed larvae harvested from cattle; similar forms appeared more slowly in larvae incubated at 31°C or 25°C. Extracts prepared from larvae after incubation at 37°C for 3–5 days or 30°C for 8 days were consistently infective for cattle. Prior storage of larvae at 14°C for up to 28 days enhanced the development of infectivity at 37°C; infectivity could still be produced after 65 days storage at 14°C but not after 76 days. Larvae released on a host transmitted B. bovis sooner if they had been incubated at 37°C for 4 days. It was concluded that the development of B. bovis to an infective stage in B. microplus is temperature dependent and does not require the stimulus of feeding by the host.     

Colonization, larval survival and epidemiology of the nematode Anguillicola crassus, parasitic in the eel, Anguilla anguilla, in Britain

In late 1987, immature Anguillicola crassus were reported for the first time in Britain from eels from two river systems. By late 1988, gravid adults were present in a number of rivers in the east of England in two discrete centres of distribution: one in East Anglia correlated with the route taken by lorries exporting eels to the continent, and one in the R. Thames correlated with the import of eels to London. The parasite was firmly established in the R. Trent, where prevalence levels reached 100% in some places. Laboratory investigations showed that adult parasites and their eggs remained viable even after infected eels had been maintained for 4 weeks in 100% sea water. Hatching of eggs declined with increasing salinity, but was not totally inhibited by sea water. Survival and infectivity of freeliving second stage larvae were maximal in natural fresh water (95% survival for 4 months, and 50% still infective to copepod intermediate hosts after 70 days), but declined in alkaline water and with increased salinity. Nevertheless, in 100% sea water, 50% of larvae were still infective after 8 days. Specificity to the intermediate host was low, and eels of all sizes could be infected. These characteristics, plus a high reproductive potential, give the parasite exceptional colonization potential and ability, enabling it to survive natural movements of eels from catchment to catchment and to increase rapidly within a new locality. The ability of free-living larvae to adhere to the substratum and survive in sea water enables them to survive in eel-transport lorries from which they will not readily be removed by flushing, the normal cleansing procedure. It is concluded that there were two separate introductions of the parasite to Britain; via the eel import trade through London, and, totally unexpectedly, via the eel export trade in lorries traversing East Anglia. The parasite is now firmly established in Britain and will continue to spread by natural movements of eels but especially by human-assisted movements of infected eels for stocking and market. This latter practice is recognized as a major factor in introducing and disseminating fish parasites.     

Phased infectivity in Heterorhabditis megidis: the effects of infection density in the parental host and filial generation

Entomopathogenic nematodes can develop through two or more generations in the cadavers of killed insect hosts. Non-feeding infective juveniles from each generation emerge and may spend prolonged periods searching for a new host. The infectivity of the infective juveniles of Heterorhabditis megidis varies with time after emergence and may not reach a maximum until several weeks have passed. 'Phased infectivity' hypotheses propose that this pattern is adaptive, tending to reduce competition in new hosts. Here we provide further evidence that infectivity is phased in H. megidis. In addition, we show that the basic pattern is modified by infection density in the parental host and by filial generation. Two general patterns were observed: first, infective juveniles that developed under the least crowded conditions (F(1) infective juveniles produced in hosts infected with 16 parent nematodes) reached maximum infectivity after only 15 days, compared to 27 or 39 days for infective juveniles that developed under more crowded conditions (F(1) produced in hosts infected with 103 or 424 parent nematodes or F(2) infective juveniles). Second, infective juveniles had lower infectivity overall when produced under the most crowded conditions (F(2) versus F(1); highest versus lowest infection density). We propose that while lower overall infectivity is a necessary consequence of limited resource availability during infective juvenile development, the difference in the timing of peak infectivity reflects a shift in the fitness gains associated with being maximally infective either 'early' or 'late'. F(1) infective juveniles emerge several days before F(2) infective juveniles, and we suggest that filial generation and infection density in the parental host function as indicators of the potential risk of competition within new hosts.