Developmental physiology of cestodes: cyclic nucleotides and the identity of putative crowding factors in Hymenolepis diminuta

Worm-conditioned saline (WCS) was prepared by incubating Hymenolepis diminuta from crowded infections for 12 hr in a balanced salt solution. The effect of the WCS on the incorporation of [3H] thymidine into DNA in the anterior regions of fresh H. diminuta was compared to effects produced by the cyclic nucleotides in the WCS. Cyclic AMP and cGMP were found in the WCS, and cGMP but not cAMP (at the concentration in WCS) caused some inhibition of DNA synthesis. For further study of the effects of cyclic nucleotides, worms were incubated with theophylline, caffeine, 3-isobutyl-1-methyl xanthine, 2-deoxy cGMP, and L-ascorbic acid, all of which produced some inhibition of [3H] thymidine incorporation. Treatment of WCS with 3',5' cyclic nucleotide phosphodiesterase abolished part of its inhibitory activity, i.e., that part presumed to be due to cGMP. When worms were incubated in the presence of succinate, acetate, D-glucosaminic acid, and cGMP simultaneously and in the concentrations each was found in the WCS, DNA synthesis was inhibited to a degree equal to that found in the WCS. Thus these substances apparently represent the putative crowding factors in the WCS. WCS prepared with worms from different population densities contained the same levels of cAMP but varied in content of cGMP, which decreased as the worm density increased. WCS prepared with patent worms contained high levels of cAMP, but the same amounts of cGMP as WCS prepared with 10-day-old worms. At least some inhibitors of cyclic nucleotide phosphodiesterase inhibited the secretion of cGMP by the worms. Levels of cGMP in the host intestine varied with the presence or absence of worms, number of worms, and area of the intestine.     

黄颡鱼肠道内寄生棘头虫位置分布的研究

长江丽棘虫和黄颡异１钩棘头虫单独在黄颡鱼肠中分布的相似性比例ＰＳ＝０．６８４。它们共同寄生在黄颡鱼肠道内,长江丽棘虫的寄生部位向前移,黄颡异钩棘头虫的寄生部位明显向后移,生态位宽度变窄,生态位重叠值下降。但是,它们的平均感染强度并未降低,种间正关联显著。     

Specific cross-immunity between Hymenolepis nana and H. diminuta: immunization with heterologous and homologous light infections

The consequences of previous and concurrent infection with two related species of cestodes, Hymenolepis nana and H. diminuta, were studied in CD1 mice. A H. diminuta infection strongly affected the establishment and the survival of a secondary H. nana egg or cyst infection administered 30 days later. An infection of 20 H. nana eggs strongly protected against a 5-cyst H. diminuta challenge, whereas an infection of 10 H. nana cysts was ineffective; 20 H. nana eggs also protected against a challenge with 5 cysts of H. diminuta administered 5 days later. No effects were observed in either parasite during a concurrent infection established by administration of cysts. An H. nana egg-infection was unable to affect the establishment of a secondary H. nana cyst-infection given 1 month later; however a significant decrease in growth was found. Similar results were found when a primary H. nana egg-infection was followed 5 days later by the homologous cyst-infection. But an infection with 5 H. nana cysts was unable to protect against a homologous challenge of 5 cysts or 200 eggs. The reciprocal cross immunity between the heterologous parasites and the failure of protection of homologous challenges are discussed in relation to light infections.     

The effect of the expulsion phase of Trichinella spiralis on Hymenolepis diminuta infection in rats.

The effect of the intestinal changes brought about by the expulsion of Trichinella spiralis in rats was studied in relation to the growth and survival of a concurrent infection with Hymenolepis diminuta, a cestode not normally rejected by the rat in low-level infections. Growth of H. diminuta was stunted in rats given T. spiralis just before, or after, infection with H. diminuta, the stunting being more pronounced when the cestode was given closer to the period of inflammation. There was no loss of the cestode from dual-infected rats and no evidence for destrobilation was found. Lower T. spiralis burdens had a correspondingly weaker effect on growth of H. diminuta, and stunting was abolished by administration of the anti-inflammatory drug cortisone acetate. It is concluded that the stunting of H. diminuta is probably due to the non-specific inflammatory component of the rat's response to T. spiralis infection.     

长江黄州江段长须黄颡鱼肠道内寄生棘头虫的生态研究

解剖检查了 133尾捕自长江黄州江段的长须黄颡鱼 ,发现寄主感染长江丽棘虫的感染率为 4 8.9% ,感染强度为 5 .4 (虫 /尾 ) ,平均密度为 2 .6 (虫 /尾 ) ,感染鲇异棘虫的感染率为 2 1.8% ,感染强度为 5 .5 (虫 /尾 ) ,平均密度为1.2 (虫 /尾 ) ,两种棘头虫在长须黄颡鱼种群中均为聚集分布 ;当两种棘头虫共同寄生在宿主肠道内时 ,其感染强度和单独感染时相比没有显著差异 ,但是 ,各自的生态位宽度均变小 ;两种棘头虫的正关联关系显著。     

Single and concurrent infections of the golden hamster, Mesocricetus auratus, with Echinostoma revolutum and E. liei (Trematoda: Digenea)

Single or concurrent infections of the intestinal trematodes Echinostoma revolutum and E. liei were studied in the golden hamster (Mesocricetus auratus). In single infections, some hamsters were fed 25 +/- 5 metacercarial cysts and others 100 +/- 25 cysts of either E. revolutum or E. liei. In concurrent infections, hamsters were fed simultaneously 20 +/- 5 metacercarial cysts of E. revolutum and 20 +/- 5 cysts of E. liei or 100 +/- 25 cysts each of both trematodes. All hamsters exposed singly to E. revolutum or E. liei were infected. In concurrent infections, 9 of 10 hamsters were infected with both species of echinostomes, and the ratio of E. revolutum to E. liei was 3:1. In single infections, 80% of the E. liei and 60% of the E. revolutum were in the posterior third of the small intestine. In concurrent infections, 80% of the E. liei were in the posterior third and 57% of the E. revolutum in the middle third of the small intestine. The histopathological response of E. liei and E. revolutum in single and concurrent infections showed erosion of intestinal villi with lymphocytic infiltration as the primary response. Extraintestinal echinostomiasis occurred in 2 of the infection groups. Differences in hemoglobin and packed cell volume occurred in the different infection groups.     

Effect of surgical alteration of the rat gastrointestinal tract on the growth and development of Hymenolepis diminuta

Eight groups of rats were used to study the involvement of the enteric (ENS) and central (CNS) nervous systems in the development of Hymenolepis diminuta using surgical intestinal transection, or CNS denervation, or both procedures. The transection procedure was used to isolate the ENS of the small intestine from either orad and/or caudal portions of the alimentary system, while the CNS denervation was used to eliminate direct visceral efferent inputs from the CNS. Nine days after the surgical procedures, all rats were infected with 35 cysticercoids of H. diminuta. On 20 days postinfection, the infection intensity, tapeworm dry weight, tapeworm morphology, intestine length, and intestinal wet weight were recorded. Only the combination of the duodenal and ileal transections with a CNS denervation reduced infection intensity and prevented the increased intestinal length normally observed in infected rats. In contrast, none of the various intestinal transection procedures alone or CNS denervation alone had any effect on the survival, ability to produce oncospheres or morphology of the tapeworms. In conclusion, tapeworm survival is decreased when both CNS and ENS inputs into the small intestine are altered or absent.     

Differential establishment and survival of Hymenolepis diminuta in syngeneic and outbred rat strains.

Experimental Hymenolepis diminuta infection was carried out in inbred strains of rats (F344/N, JAR-2, LOU/M, TM, DA and DA-bg/bg) and outbred Wistar rats. All strains became infected with this cestode, but clear strain-dependent variation in the susceptibility to H. diminuta infection was observed. Marked differences in worm persistence and worm weight were found at 6 weeks post-infection in TM and DA rats. These strains would be useful to clarify the interactions between H. diminuta and its rat host.     

Bioecology of the parasites of high altitude homeothermic host-parasite systems. II. Infections by strobilocerci of Hydatigera balaniceps (Hall), H. laticollis (Rudolphi) and H. parva (Baer) in relation to sex, size and weight of Indian house rat

The influence of sex, size and weight of the Indian house rat (Rattus rattus) on concurrent liver infections with Hydatigera balaniceps (Hall), H. laticollis (Rudolphi) and H. parva (Baer) has been discussed. A significant inverse relationship was recorded between cestode infection and rat size. Parasitization index and mean worm burden-host weight ratio decreased with increase in rodent weight.     

Biochemical effects of thiabendazole and cambendazole on Hymenolepis diminuta (Cestoda) in vivo

An investigation of the chemotherapeutic and biochemical effects of two benzimidazole anthelmintics, thiabendazole (TBZ) and cambendazole (CBZ), on Hymenolepis diminuta in experimentally infected rats is reported. Thiabendazole was active against H. diminuta at a relatively high dosage. A single oral dose of TBZ at 250 mg/kg body weight on day 15 of infection eliminated 100% of the tapeworms as determined at necropsy 5 days after treatment. The chemotherapeutic actions of TBZ on H. diminuta were accompanied by marked changes in worm weight and chemical composition. Tapeworms recovered from rats that had received a therapeutically effective dose of TBZ 24 hr earlier were significantly smaller and contained much less glycogen (as a percent of the wet weight) than worms from unmedicated controls. Protein concentrations increased in TBZ-treated worms and at a rate sufficient to offset the decline in glycogen concentration. Glycogen/protein ratios in TBZ-treated worms were significantly lower than the corresponding control values. Cambendazole proved to be five times more potent than TBZ against H. diminuta and produced the same basic changes in worm weight and chemical composition within 18 hr of treatment of the host. Administration of a single oral dose of TBZ or CBZ to the host produced in H. diminuta another change, the onset of which coincided with, or preceded, the gross alterations in worm weight and chemical composition. That change, observed in in vitro studies carried out 14 hr after treatment, revealed that tapeworms from drug-treated rats absorbed and metabolized much smaller quantities of exogenous glucose than did the controls, and the ability of the worm to accumulate glucose against a concentration difference was significantly depressed.     

Description and pathology of Neoechinorhynchus idahoensis n. sp. (Acanthocephala: Neoechinorhynchidae) in Catostomus columbianus from Idaho.

Neoechinorhynchus idahoensis is described from Catostomus columbianus caught in the Salmon River, Stanley Basin, Idaho. The new species is closest to Neoechinorhynchus venustus Lynch, 1936, but is distinguished from it by its smaller and variably structured eggs, anterio-dorsal trunk hump, bent and posteriorly notched proboscis receptacle, and larger proboscis, proboscis receptacle, and hooks. It is distinguished also from 2 other species of Neoechinorhynchus with proboscis hooks in middle and anterior circles about equally large and from 7 other species having lemnisci greatly unequal in length. Histopathology of host tissue showed limited host response exemplified by epithelial damage and hemorrhaging at point of proboscis attachment with subsequent macrophage and other phagocytic cell migration. The proboscis extended through the host epithelium into the submucosa with limited hemorrhaging at the point of attachment. Unorganized collagenous fibers were present. The lumen of the host intestine was obstructed, and compressed villi were present. The trunk of the worm damaged intestinal epithelium near the crypts, causing localized inflammation. The caryophyllaeid cestode Isoglaridacris calentinei Mackiewicz, 1974, was present in concurrent infections of C. columbianus.     

Adsorption of bile salts by the cestodes, Hymenolepis diminuta and H. microstoma.

The accumulation of purified sodium taurocholate (NaTC) and sodium glycocholate (NaGC) by Hymenolepis diminuta and Hymenolepis microstoma (Cestoda: Cyclophyllidea) was determined using radioactive bile salts. H. diminuta reached equilibrium levels of approximately 120 nmoles NaTC/g dry wt and 300 nmoles NaGC/g dry wt. Presentation of the bile salts in mixed micelles with 0.35 mM oleic acid did not alter these values. With H. microstoma, the maxima were 195 nmoles NaTC/g dry wt and 614 nmoles NaCG/g dry wt. These values were similarly unaffected by the addition of 0.35 mM oleic acid to the micelles. Equilibrium values of this magnitude, in media containing as much as 25 or 30 mM bile salt, and the maintenance of this level during incubations of 15 to 60 min eliminated the possibility that the accumulation was by diffusion or by any form of mediated transport into the worm. The accumulation on NaTC by H. diminuta was [Na+] independent, and insensitive to ouabain, DNP, and high [K+]. These observations, the maintenance of different levels of NaTC and NaGC, and the failure of the 2 bile salts to compete indicated that there was no active excretion mechanism operating in a fashion similar to the active transport of bile salts in the vertebrate small intestine. It was concluded that the accumulation of NaTC by H. diminuta was actually adsorption to the tegument. Comparable, although more limited, experiments extended this conclusion to the accumulation of NaGC by H. diminuta and of NaTC and NaGC by H. microstoma. It is suggested that bile salt monomers, rather than intact micelles, adsorb to specific loci on the tegument.     

Shape variation of cysticercoids of Hymenolepis diminuta (Cyclophyllidea) from fed, partially fed, and fasted Tribolium confusum (Coleoptera)

Quantitative studies of a crowding effect on cysticercoids of Hymenolepis diminuta in the intermediate host are few and limited in scope. In this study, we developed a technique to rapidly collect morphological information on large numbers of parasites, and verified the utility of geometric models for simple and accurate estimation of cysticercoid size for quantitative studies. These models were tested using measurements from 4,899 H. diminuta obtained from 666 Tribolium confusum exposed 1-4 wk previously. Length, width, and depth of the body and cercomer (when present) can be used in conjunction with these models to provide the most accurate estimation of parasite size. However, parasite body length alone can be used, with adjustment for effects of host diet and infection intensity, to predict the remaining measurements in incomplete specimens. Parasites that developed in higher intensity infections, or in hosts with reduced food intake, were narrower and had a proportionately shorter cercomer. Host age, sex, and mating status, and parasite age also had statistically significant, but small-magnitude, effects on parasite shape.     

Secondary infections of Hymenolepis diminuta in mice: effects of varying worm burdens in primary and secondary infections.

In one (1 c) and six (6 c) cysticercoid primary infections of Hymenolepis diminuta in NIH (inbred) and CFLP (outbred) male mice 6 +/- 1 weeks old greater than 85% of the worms established but were rejected (destrobilated or expelled) subsequently. Rejection occurs more quickly in 6 c infections than in 1 c infections. Considerable worm growth occurs in 1 c and 6 c primary infections but worms from 6 c infections weighed less than worms from 1 c infections on all days studied. Expulsion of H. diminuta does not occur more rapidly in secondary infections than in primary infections; loss of 6 c secondary worms occurs at the same rate as 6 c primary worms but 1 c secondary worms survive longer than 1 c primary worms. Although worms are not lost more quickly in secondary than in primary infections, they are affected at an early age by the immune response which stunts their growth. Increasing the intensity of primary and secondary infections increases the severity of stunting of secondary worms. The results are discussed and it is suggested that immune responses to Hymenolepis spp. in rodents are common but that thresholds of worm numbers exist below which appreciable worm loss does not occur. Stunting due to crowding, which generally is attributed to inter-worm competition, may be in part immunologically mediated. For future immunological studies attempting to induce secondary responses to H. diminuta in mice, worm growth, not survival, is the criterion to evaluate.     

Hymenolepis diminuta: effects of decerebration on thermal response behavior.

Hymenolepis diminuta propels itself with unidirectional peristaltic-like waveforms. When intact adult H. diminuta are placed in a thermal gradient, with the anterior proglottids hot relative to the posterior proglottids, the worms migrate up the gradient toward the hot side. When the anterior is cold, relative to the posterior, the worms moved slightly or little. These behaviors in a thermal gradient represent true thermokinetic responses for an organism with undirectional locomotion. Removal of the scolex, containing the worm's cerebral ganglia, did not significantly alter these thermal responses. These data suggest that the peripheral nervous system is capable of integrating sensory input over the length of the strobila and coordinating locomotory behavior, in the absence of the central nervous system.     

In vivo effects of putative crowding factors on development of Hymenolepis diminuta.

During in vitro incubation, Hymenolepsis diminuta secretes substances into the medium that inhibit DNA synthesis in the germinative region of freshly isolated, uncrowded worms. Of the many substances that are released by H. diminuta into the medium, earlier studies indicate that only succinate, acetate, glucosaminic acid, and cGMP are responsible for the inhibition. In the present report, effects of these putative crowding factors on worm development in vivo were examined. At 7 days postinfection the proximal end of the host's intestine was catheterized and perfused with test solution. The test solution contained 28 nM cGMP, 250 microM glucosaminic acid, 120 mM succinate, and 40 mM acetate. The solution was perfused by a peristaltic pump at a rate of 50 ml/day. At 2 wk postinfection, worms were recovered for subsequent analysis. Worms developing in the presence of crowding factors were 53% less in wet weight than control worms. Carbohydrate concentrations in worms from experimental groups were not different from those in control groups; therefore, the inhibition in growth was probably not due to carbohydrate deprivation. Worms from experimental groups had fewer immature, mature, and gravid proglottids than did worms from control groups. The results are consistent with the hypothesis that the tested substances, which inhibit DNA synthesis in H. diminuta in vitro, are a part of the cause of the crowding effect in vivo.     

Hymenolepis citelli, H. diminuta and H. microstoma: immunoglobulin-containing cells in the lamina propria of the mouse gut during primary and secondary infections

The indirect immunofluorescent technique was used to determine the occurrence of IgA, IgM and IgG1 immunoglobulin-containing cells in local intestinal mucosal immune responses to Hymenolepis citelli, H. diminuta and H. microstoma infections in mice. In the intestinal lamina propria of H. citelli and H. diminuta infected mice there was no increase in the mean numbers of immunoglobulin-containing cells when compared with uninfected control mice, but there was in H. microstoma infected mice. The numbers of IgG1- positive cells in both infected and uninfected mice were very small relative to IgA and IgM immunocytes. The distribution of immunocytes in the lamina propria of infected and uninfected mice was essentially similar and the localization of isotypes in duodenal sections showed no immunoglobulins in the villous epithelial cells. There was also no marked difference between primary and secondary infections indicating that immunoglobulin-containing cells play no major role in functional immunity against hymenolepid infections in the mouse. The presence of IgA and IgM was also demonstrated on the tegument of the tapeworms, although the distribution was patchy and more abundant on H. microstoma than on H. diminuta or H. citelli. The time of appearance of both isotypes was latest on H. citelli.     

Ileal mucosal mast cell, eosinophil, and goblet cell populations during Hymenolepis diminuta infection of the rat.

The population dynamics in the enteric connective tissues of eosinophils, mucosal mast cells (MMC), and in the mucosal epithelium of goblet cells were examined morphometrically in fixed ileal tissue of outbred Sprague Dawley rats during the first 32 days of infection with the tapeworm Hymenolepis diminuta. MMC and eosinophils were present in the lamina propria and submucosa; however, only eosinophils were also present in the muscularis externa. Eosinophilic infiltrate was first observed in the lamina propria at 15 days postinfection (dpi) and the numbers of eosinophils remained elevated through 32 dpi. Initial mucosal mastocytosis was detected on 6 dpi and MMC numbers continued to rise over the study period without reaching a plateau. Goblet cell hyperplasia occurred only at 32 dpi. In contrast to some intestinal nematode infections where these same 3 cell types are associated with the host's expulsion responses, H. diminuta is not lost by a rapid host response in the outbred Sprague Dawley rat strain used in these experiments. We suggest that either the induction of hyperplasia of these host effector cells in ileum tissue during H. diminuta infection is not capable of triggering parasite rejection mechanisms, or the function of the induced hyperplasia is necessary for some as yet unassociated physiological or tissue architecture change in the host's intestine.     

Water balance and its relation to fermentation acid production in the intestinal parasites Hymenolepis diminuta (Cestoda) and Moniliformis moniliformis (Acanthocephala).

Water balance and its relation to carbohydrate metabolism was examined in Hymenolepis diminuta in parallel with the putative osmoconformer Moniliformis moniliformis. Worms were removed from rat intestines, weighed, and incubated (37 C) 1 hr in rat serum and various salines, some with mannitol to vary osmotic concentration from 150 to 400 mOsm/L. Worms were removed at 15-min intervals, weighed, and returned to the test solution. Rat serum and a Ringer's saline (pH 7.4 and 300 mOsm/L) with or without 5 mM glucose were isotonic to M. moniliformis, which behaved like an osmometer, shrinking, or swelling in proportion to external osmotic changes. Hymenolepis diminuta rapidly lost 20-25% wet weight in these solutions and regained lost water when 5 mM glucose was added to the saline. Tapeworms maintained constant body weight between 210 and 335 mOsm/L, but they rapidly gained or lost water outside of this range. Glucose metabolism and uptake of [3H]glucose from the medium increased progressively between 210 and 310 mOsm/L, whereas uptake rates of [3H]leucine, 22Na+, and 36Cl- were not affected. Unbuffered saline (initial pH 6.5 and 300 mOsm/L) had a lower pH (5.0) and higher osmolality (307 mOsm/L) after a 1-hr incubation with tapeworms. Such saline was less hypertonic than unconditioned saline to freshly obtained worms. A Ringer's saline (300 mOsm/L) containing 50 mM acetate- was also hypertonic (greater than 20% weight loss) to tapeworms at pH 7.4, but it was hypotonic (greater than 20% weight gain) at pH 5.0. Isotonicity at 300 mOsm/L was achieved with pH 5.0 and 20 mM acetate-, the approximate pH and fermentation acid concentration in an infected rat intestine. Rats infected with tapeworms (12 days old) were fasted for 2 days. Starved worms were smaller but had the same percentage of body water and internal osmolality as controls. These results show that H. diminuta can regulate its body water content and that water balance is closely related to the fermentation acid concentration and pH of the bathing medium.     

Some influences of population density on Hymenolepis diminuta in rats.

When measured 56 days postinfection the length, wet weight and dry weight of Hymenolepis diminuta were all found to decrease with increasing number of cysticercoids given up to 20. The mean position of the worms in 10, 12 and 20 worm infections is significantly posterior to that of 1, 2 and 5 worm infections and the worms are attached over a wider area of the intestine. Egg production by the worms was followed up to day 56 postinfection; the number of eggs produced per worm and even per rat decreased with increasing population density. Thus the best way to get most eggs and to maintain the parasite in the laboratory is to have rats infected with only one tapeworm. Rats given 1-20 cysticercoids showed a mean recovery of 100-65%, while rats given 40-200 cysticercoids showed a mean recovery ranging from 13 to 2%. In addition to 'normal' worms, defined as worms greater than 10 mm, small, most probably destrobilated, worms were found. In the 50 and 100 cysticercoid infections, worm recoveries were, respectively, 8% 'normal', 16% small, and 2% 'normal', 5% small. From the significantly lower recovery from heavy infections it is concluded that a deleterious factor is operating during the 8 weeks after the infection.