Differences in adult egg output of Schistocephalus solidus from singly- and multiply-infected sticklebacks

In this study, we recovered Schistocephalus solidus plerocercoids from singly and multiply infected three-spine sticklebacks, Gasterosteus aculeatus, transferred them individually to in vitro culture conditions, and quantified their lifetime egg output. We found a significant difference in the relationships between plerocercoid mass and lifetime egg output for parasites recovered from singly and multiply infected sticklebacks. Although egg output was strongly and positively related to plerocercoid mass amongst worms from singly infected fish, for those recovered from multiply infected sticklebacks the relationship was marginally nonsignificant and negative, with small worms achieving high levels of egg production. We suggest 2 hypotheses that may explain differences in the egg production of plerocercoids from singly and multiply infected fish. One possibility is that smaller plerocercoids in asymmetric multiple infections develop precocially, in response to host manipulation strategies of larger worms that decrease survival prospects of the host. Alternatively, small worms in singly infected sticklebacks may be prevented from becoming sexually mature because they face energetic constraints associated with having to overcome the host's immune response alone. We discuss our results in terms of recent studies examining strategic egg production in helminths.     

Development of the plerocercoid I of Ophiotaenia europaea in reptiles

The existence of a two-host life-cycle in ophiotaeniid proteocephalideans was tested experimentally using Ophiotaenia europaea as a model. Three species of reptiles, Natrix natrix, Natrix tessellata and Lacerta viridis, were fed with experimentally infected copepods containing a large number of infective plerocercoids I. A few plerocercoids, most of which were dead, corresponding morphologically to the plerocercoid II developmental stage of O. europaea, were found encysted in the intestinal wall of N. natrix (8 days p.i.), N. tessellata (5 and 150 days p.i.) and L. viridis (40 days p.i.), while no plerocercoids or adult worms were recovered from their intestines. The results indicate that the infective plerocercoid I of O. europaea cannot undergo further development when ingested directly by the final host (a reptile), and that environmental temperature stimuli cannot initiate a reverse plerocercoid migration to the gut followed by strobilization.     

Some (worms) like it hot: fish parasites grow faster in warmer water,and alter host thermal preferences

Elevated environmental temperatures associated with anthropogenic warming have the potential to impact host‐parasite interactions, with consequences for population health and ecosystem functioning. One way that elevated temperatures might influence parasite prevalence and intensity is by increasing life cycle completion rates. Here, we investigate how elevated temperatures impact a critical phase of the life cycle of the bird tapeworm Schistocephalus solidus – the growth of plerocercoid larvae in host fish (three‐spined sticklebacks Gasterosteus aculeatus). By 8 weeks post‐infection, plerocercoids recovered from experimentally infected sticklebacks held at 20 °C weighed on average 104.9 mg, with all exceeding 50 mg, the mass considered consistently infective to definitive hosts. In contrast, plerocercoids from sticklebacks held at 15 °C weighed on average 26.5 mg, with none exceeding 50 mg. As small increases in plerocercoid mass affect adult fecundity disproportionately in this species, enhanced plerocercoid growth at higher temperatures predicts dramatically increased output of infective parasite stages. Subsequent screening of thermal preferences of sticklebacks from a population with endemic S. solidus infection demonstrated that fish harbouring infective plerocercoids show significant preferences for warmer temperatures. Our results therefore indicate that parasite transmission might be affected in at least two ways under anthropogenic warming; by enhancing rates of parasite growth and development, and by increasing the likelihood of hosts being able to seek out proliferating warmer microhabitats. Furthermore, our results suggest the potential for positive feedback between parasite growth and host thermal preferences, which could dramatically increase the effects of even small temperature increases. We discuss the possible mechanisms underpinning our results, their likely ecological consequences and highlight key areas for further research.     

Survey of Spirometra erinaceieuropaei in frogs in Taiwan and its experimental infection in cats

Eighteen of 56 (32.1%) wild Rana limnocharis from central and south Taiwan were found to contain plerocercoids of Spirometra erinaceieuropaei. This is the first report of S. erinaceieuropaei infections in frogs in Taiwan, with the plerocercoids being recovered from the thigh and back muscles or under the skin. Other species of frogs examined, including nine wild R. latouchii, one wild Buergeria robustus and 110 cultured R. rugulosa were free of infection. The plerocercoids were orally inoculated into four cats; three of which were each given a single plerocercoid and one a dose of three plerocercoids. Daily faecal examination showed that two cats started shedding eggs of S. erinaceieuropaei on day 8 postinfection (PI) and the other two on day 10 PI. The highest eggs per gram and eggs per day for a single worm was found to be 428,000 and 14,416,000 respectively. Only the cat inoculated with three plerocercoids shed proglottids in its faeces during the 2 month observation period.     

Host specificity of adult versus larval cestodes of the elasmobranch tapeworm order Trypanorhyncha

Host specificity between the adult and final larval stages (plerocercus, plerocercoid, or merocercoid) of a diversity of trypanorhynch species was compared using the host specificity index (HS s). Index values were generated for a total of 63 species representing all five trypanorhynch superfamilies and 11 families. Host specificity of both adults and final larvae was found to be widely variable among species, ranging from very high (oioxenous) to very low (euryxenous) for both stages. However, in general, host specificity was highest for the adult stage in the definitive host (mean HS s=3.86) and lowest for the final larval stage in the second intermediate host (mean HS s=6.29). This difference was found to be significant using the Wilcoxon signed-rank test. Limited data available for procercoids in the first intermediate host suggest that this stage exhibits a degree of specificity intermediate between that of the former two stages (mean HS s=4.23). No taxonomic trend was seen. Species with a plerocercoid final larval stage (mean HS s=8.62) were significantly less host-specific than those with plerocerci or merocercoids (mean HS s=5.56). This result may reflect the use of paratenic hosts by species possessing the relatively more resilient plerocercoid as their final larval stage. These results provide an example of how HS s can be used to compare levels of host specificity, in this instance, among stages of polyxenous life cycles. They also emphasise the importance of articulating the life cycle stage under consideration when general statements are made about host specificity.     

Age of adult worms of Echinostoma caproni does not affect development of miracidia

The effect of the age of adult Echinostoma caproni on egg development was studied. The percentage of fully developed miracidia was determined in eggs derived from adult worms obtained from laboratory mice at 2, 4, 6, and 8 wk postinfection (PI). Regardless of the age of worms from which the eggs were obtained, the percentage of fully developed miracidia was always >90%, and 60-80% of the eggs hatched. Several previous studies have shown that eggs derived from 2- to 4-wk-old E. caproni yielded miracidia that infected Biomphalaria glabrata snails. Results of the present study on E. caproni were in marked contrast to previous results with Echinostoma friedi, for which viable eggs were not obtained at 2 and 3 wk PI and maximal infectivity of miracidia in snails was obtained from eggs derived from worms collected at 8 and 9 wk PI. Further studies are needed to determine if the egg viability of other species in the "revolutum" group follow that of E. caproni or E. friedi.     

Loss of Hymenolepis diminuta from the rat

One, 5, 15 and 30 worm infections of Hymenolepis diminuta were established in juvenile or adult male (Hooded Rowett or Sprague-Dawley) rats. Worm numbers and weight, and egg output were determined from day 15 to day 85 post infection. Gradual worm loss occurred only from 15 and 30 worm infections. In 5, 15 and 30 worm infections worm weight decreased from day 19 to day 50 but no weight loss occurred in single worm infections. The size range of individual worms from a multiple infection of a single rat increased markedly following infection. Adult rats showed a greater worm loss and harboured smaller worms than juvenile rats. The data will fit either a competitive or an immunological model.     

Infectivity, growth, and distribution of Echinostoma caproni (Trematoda) in the ICR mouse

Host-parasite interactions of the intestinal trematode Echinostoma caproni were studied in ICR laboratory mice. All of 40 mice, each fed 25 metacercarial cysts of Echinostoma caproni, were infected 1-20 wk postinfection (PI) with a mean of 17.2 worms/host. At 24 and 29 wk PI only 2 of 6 mice (33%) were infected, with a mean of 4.2 worms/host. Mean body area of worms increased rapidly to about 5 mm2 by week 2, increased less rapidly to 8.8 mm2 by week 12, plateaued until week 24, and then declined. Mean dry weight of worms increased rapidly to about 0.5 mg by week 2, less rapidly to 1.4 mg by week 12, and then plateaued until week 24 PI. From 1 to 8 wk PI most worms localized in the jejunum and ileum; later most worms were in the jejunum and duodenum. Considerable differences were seen in the growth and distribution of E. caproni in the ICR mouse compared with previous studies on this echinostome species in the NMRI mouse.     

Development of Taenia asiatica cysticerci to infective stage and adult stage in Mongolian gerbils

The development of metacestodes and adult worms of Taenia asiatica in Mongolian gerbils (Meriones unguiculatus) were observed. Cysticerci were recovered from gerbils subcutaneously injected with hatched oncospheres. The recovery rate ranged from 0.1 to 3.2%. No cysticerci were recovered from the orally inoculated gerbils. The infectivity of the cysticerci recovered at 48 weeks post-infection was evaluated. Tapeworms were recovered on day 14 post-infection from the small intestine of 5 of 11 gerbils, with a recovery rate of 27% (6 worms recovered/22 worms inoculated). Three and four adult worms were recovered from two human volunteers who ingested five cysticerci after 4 months post-infection. In worms recovered from gerbils, segmentation and genital primordia in the posterior proglottids and hooklets in the residual rostellum were observed. The results indicate that gerbils can serve as an alternative intermediate host and that partial development of the adult worm stage occurs in gerbils.     

Comparison of microthrix ultrastructure and morphology on the plerocercoid and adult scolex of Calliobothrium cf. verticillatum (Tetraphyllidea: Onchobothriidae)

The distal bothridial surfaces of adult triloculate onchobothriids are covered with short structures that have been tentatively classified as very short filitriches, but this hypothesis has never been tested. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to investigate microthrix morphology in the plerocercoid and adult forms of Calliobothrium cf. verticillatum, a triloculate onchobothriid tapeworm from Long Island Sound (Connecticut). Plerocercoids of C. cf. verticillatum were collected from the anterior midgut ceaca of Pagurus pollicaris Say, 1817 (flat-clawed hermit crab), and adults were collected from the spiral intestine of the dusky smooth hound Mustelis canis (Mitchell, 1815). Two plerocercoids and 2 adults were examined using SEM; 2 plerocercoids and 2 adults were examined using TEM. Microthrix distribution and morphology (including measurements of total length, base length, shaft length, and base width) were investigated on all surfaces of the plerocercoid and adult scolex. Slender filitriches and large bladelike spinitriches were observed extending from the tegument of plerocercoid and adult forms. The filitriches were found to have significantly narrower bases than the spinitriches (65-167 nm vs. 466-1,936 nm, respectively). The scolex proper of the plerocercoid and adult forms were found to have filitriches of medium-length and bladelike spinitriches. The distal bothridial surfaces differed dramatically in microthrix morphology between plerocercoid and adult forms; on the distal surfaces of the plerocercoids were long filitriches and bladelike spinitriches. However, the distal surfaces of the adults had short structures (previously hypothesized to be short filitriches) and a few bladelike spinitriches. Serial transverse sections revealed that the short structures on the distal bothridial surfaces of the adults were homologous with filitriches. They included all of the structural components of a filithrix as well as a base width that conformed to the filitriches found on other surfaces. The bothridial margins of the plerocercoid and adult forms had a microthrix pattern similar to that seen on the proximal bothridial surfaces except that the filitriches on the margins were significantly longer than those found anywhere else on the bothridia. The most dramatic difference between the plerocercoid and adult forms occurred on the distal bothridial surfaces, where the filitriches of the adult cestodes were significantly shorter and narrower, and the spinitriches were almost entirely lacking.     

Worm morphology of Schistosoma japonicum using confocal laser scanning microscopy

Male and female Schistosoma japonicum worms have dissimilar appearances in their final host. In this study, a morphometric and morphological assessment of whole worms derived from unisexual and mixed infections in mice was conducted using confocal laser scanning microscopy. Worms from mixed infections showed significant morphological changes between 15 and 25 days post-infection (PI). On the fifteenth day PI, 33% of males had formed the conspicuous gynecophoric canal, but only 8% of them had testicular lobes containing a few germinative cells; 13% of females had incipient ovaries with a few immature ovarian cells inside. On the twentieth day PI, the testicular lobes contained more germinative cells in all male worms, while female worms presented vitelline glands. On the twenty-fifth day PI, more germinative cells were observed in the male testicular lobes, and differentiated cells were present in the female ovaries. All worms had fully developed reproductive organs from 30 days PI onwards. Morphometric analysis showed significant differences between mixed and unisexual infections at 35 days PI. Ovaries of worms from unisexual infections contained cells in one stage of maturation and vitelline glands had undifferentiated cells. Our study of S. japonicum provides a detailed comparison of different morphological traits from worms of mixed and unisexual infections throughout development.     

Inflammatory cell stimulation and wound healing in Sphaeridiotrema globulus experimentally infected mallard ducks (Anas platyrhynchos)

Thirty laboratory-reared mallard ducks (Anas platyrhynchos) were experimentally infected with Sphaeridiotrema globulus. Host cell-mediated immunity and wound healing in S. globulus infected ducks were evaluated by gross and histological examination. Establishment, location, and life span of S. globulus differed from previous reports of sphaeridiotremiasis in both naturally and experimentally infected waterfowl. No worms were recovered from the ceca, and worm migration occurred anterior to the ileo-cecal valve with greater dispersion (less worm crowding) at higher rates of infectivity. Parasite death and host lesion resolution were evident at days 8 to 10 postinfection (PI) in ducks fed a moderate dose (200 metacercariae, group A) with a 5% mean parasite recovery rate. Host death occurred at days 3 to 6 in ducks fed a high dose (550 metacercariae, group B) with a 16% mean parasite recovery rate. Mast cells increased significantly (P less than 0.005) in group A ducks from days 4 to 10 PI. Eosinophil proliferation was greater in group B than in group A on day 4 PI, but comparatively fewer eosinophils were identifiable in group B ducks on day 6 PI.     

Schistocephalus solidus and Ligula intestinalis: Pinocytosis by the tegument

Using ruthenium red as a macromolecule, endocytosis was demonstrated in the plerocercoid of Ligula intestinalis and adult Schistocephalus solidus. Uptake, transport across the tegument, and exocytosis across the basal plasma membrane occurred within 6 min. The types of vesicles in the tegument of L. intestinalis are redescribed and their former classification is modified. The vertical and longitudinal distribution of pinosomes in the tegument of adult S. solidus and L. intestinalis plerocercoids were determined. The possible role of macromolecular uptake in the economy of pseudophyllidean tapeworms is discussed with particular reference to growth and defence of an unencysted larval stage in the tissues of the intermediate host.     

Strongyloides ratti: the role of enteral antigenic stimuli by adult worms in the generation of protective immunity in rats

Immunogenicity of adult Strongyloides ratti was studied in rats. Immunization of rats by intraduodenal implantation of adult worms could completely inhibit the egg production and hasten the expulsion of challenged worms which were developed from subcutaneously inoculated L3 or were implanted intraduodenally as adults. Enteral immunization by intraduodenal implantation of adult worms was, however, not able to affect the esophageal larval output of the challenge infection with L3. In contrast to enteral immunization with adult worms, immunization by full sequence of a primary infection or by a combination of drug-abbreviated infection and adult worm implantation could suppress the esophageal larval output of the challenge infection. The relationship between the host defense mechanism and the life cycle of S. ratti is discussed.     

Physiological specificity of the interaction between Traenophorus nodulosus (Cestoda) and its host--fishes

The influence of the T. nodulosus plerocercoid invasion on the liver of the intermediate fish hosts (perch) was studied. The water, protein, glycogen and glucose contents was examined in adult worms inhabiting the intestine of definitive hosts (pike), the rate of glucose accumulation along the strobile of parasite and an ability of adult worms to hydrolyze the main nutrients was also investigated. As a result of the plerocercoid infection, the glucose and glycogen contents in the liver of perch juveniles decreased, while the wet weight of liver increased. It was found, that the water and protein contents from the anterior to posterior section of strobile in T. nodulosus adults was not significantly different, while the glucose contents and the rate of its accumulation decreased. The data confirming the possibility to realize the final stages of protein and carbohydrate hydrolisis with the participation of membrane digestion on the surface of helminths and proving the ability of cestodes. The problems of relations of T. nodulosus with intermediate and definitive hosts at the level of the whole organisms are discussed.     

The consequences of short-term grazing of bioactive forages on established adult and incoming larvae populations of Teladorsagia circumcincta in lambs

The objective of this study was to investigate the consequences of short-term grazing on bioactive forages on (i) the viability and fecundity of established adult Teladorsagia circumcincta population and (ii) the establishment and development of incoming T. circumcincta infective larvae. Forty-eight, parasite naive, 3-month old, grazing lambs were artificially infected with 8000 infective larvae of T. circumcincta on day 1 of the experiment. On day 21 p.i., lambs were allocated to one of three bioactive forage grazing treatments; chicory (Cichorium intybus), sulla (Hedysarum coronarium), lotus (Lotus pedunculatus), and the control grass/clover (Lolium perenne/Trifolium repens) forage. On day 28 of the experiment a second dose of 8000 T. circumcincta infective larvae was administered to the lambs to investigate the effects of forages on the ability of infective larvae to establish within the host. All animals were slaughtered for worm recovery on day 35, while liveweight gain, feacal egg counts (FEC) and total worm egg output were monitored regularly throughout the experiment. Although FEC or total egg output were similar among the groups, adult worm burdens at slaughter were significantly affected (P<0.05) by forage treatment during the 2 week grazing period. Lambs grazing chicory had the lowest adult worm burdens and significantly lower numbers of male worms compared to those grazing on grass/clover (P<0.01), while the lambs grazing on sulla or lotus had similar adult populations to grass/clover fed animals. The results from the worm recoveries of the second dose (immature worm burdens) were affected by physiologically and/or immunologically mediated mechanisms, which reduced larval establishment in all treatments. Nevertheless, immature worm burdens at slaughter were similar between chicory, sulla and grass/clover group, while the immature worm recoveries from the lotus group were significantly higher (P<0.05) compared to those from lambs grazing grass/clover. Overall, the results of the present study support the view that chicory can be a promising candidate species in pasture management practices to control T. circumcincta burdens.     

Significance of the distribution of 57Co-vitamin B12 in Spirometra mansonoides (Cestoidea) during growth and differentiation in mammalian intermediate and definitive hosts

The distribution of labeled cyanocobalamin (CN-[57Co]Cbl = [57Co]-vitamin B12) in pleurocercoids and adult tapeworms of Spirometra mansonoides was studied during development in mice 22 days days PI, respectively. Plerocercoid scolices, obtained by cutting away their bodies or by in vitro enzymatic dissolution of the bodies, were pulsed with CN- magnitude of 57Co Cbl for 1h at 37 degrees C and reimplanted subcutaneously into mice or given per os to cats. In regenerated plerocercoids, the highest concentration of magnitude of 57Co Cbl occurred in the scolex and then decreased posteriorly in the newly-formed tissues of the body. Approximately 60% of the total magnitude of 57Co Cbl present remained concentrated in the scolex following body regeneration plerocercoids and adult tapeworms of Spirometra mansonoides was studied during development in mice 22 days post-infection (PI) and in cats 16 days PI, respectively. Plerocercoid scolices, obtained by cutting away their bodies or by in vitro enzymatic dissolution of the bodies, were pulsed with CN-[57Co]Cbl for 1 h at 37 degrees C and reimplanted subcutaneously into mice or given per os to cats. In regenerated plerocercoids, the highest concentration of [57Co]Cbl occurred in the scolex and then decreased posteriorly in the newly-formed tissues of the body. Approximately 60% of the total [57Co]Cbl present remained concentrated in the scolex following body regeneration for up to 109 days PI. This high [57Co]Cbl concentration in the plerocercoid scolex was bound to protein and appears to be maintained by a complex homeostatic mechanism in association with directional transport of [57Co]Cbl to the scolex with ultimate depletion along the length of the body.(ABSTRACT TRUNCATED AT 250 WORDS)     

Susceptibility of Laboratory Rodents to Trichinella papuae

Members of the genus Trichinella are small nematodes that can infect a wide range of animal hosts. However, their infectivity varies depending on the parasite and host species combination. In this study, we examined the susceptibility of 4 species of laboratory rodents, i.e., mice, rats, hamsters, and gerbils to Trichinella papuae, an emerging non-encapsulated Trichinella species. Trichinella spiralis and Trichinella pseudospiralis were also included in this study for comparison. Fifteen animals of each rodent species were infected orally with 100 muscle larvae of each Trichinella species. Intestinal worm burden was determined at day 6 and 10 post-inoculation (PI). The numbers of muscle larvae were examined at day 45 PI. The reproductive capacity index (RCI) of the 3 Trichinella species in different rodent hosts was determined. By day 6 PI, 33.2-69.6% of the inoculated larvae of the 3 Trichinella species became adult worms in the small intestines of the host animals. However, in rats, more than 96% of adult worms of all 3 Trichinella species were expelled from the gut by day 10 PI. In gerbils, only 4.8-18.1% of adult worms were expelled by day 10 PI. In accordance with the intestinal worm burden and the persistence of adults, the RCI was the highest in gerbils with values of 241.5±41.0 for T. papuae, 432.6±48 for T. pseudospiralis, and 528.6±20.6 for T. spiralis. Hamsters ranked second and mice ranked third in susceptibility in terms of the RCI, Rats yielded the lowest parasite RCI for all 3 Trichinella species. Gerbils may be an alternative laboratory animal for isolation and maintenance of Trichinella spp.     

Lesion induction by the plerocercoid Lacistorhynchus tenuis (Cestoda) and wound healing in the striped bass Morone saxatilis (Walbaum)

San Francisco Bay-Delta striped bass, Morone saxatilis (Walbaum), form open lesions in response to a plerocercoid infection of Lacistorhynchus tenuis (Van Beneden, 1858) (Cestoda: Trypanorhyncha). Laboratory infection experiments showed that striped bass can be infected with the plerocercoids by ingesting infected copepods. Histological sections indicated that a cellular host response was mounted early in the infection period, and that despite the leucocytic infiltration the parasites continued to develop. However, at 3 months post-infection some of the plerocercoids began to degenerate, and lesions formed at this time and 14 months post-infection. Open lesions in adult striped bass collected from the field took 2 months to heal and were detectable for at least 22 months. Regeneration of the muscle tissue did not occur although the wound completely healed externally.