Hymenolepis diminuta: Worm loss and worm weight loss in long-term infections of the rat

Infections of one and two Hymenolepis diminuta established in newly weaned rats continued to grow for the duration of the experiment (238 days), whereas infections of 5 worms per rat became asymptotic around Day 55 postinfection and remained at or below this level thereafter as shown by biomass and mean weight per worm measurements. Infections of 50 worms established in newly weaned rats became asymptotic around Day 28 postinfection and thereafter worms were lost from the rats. Initially the biomass fell with the loss of worms, but by Day 56 a new lower biomass persisted for the remainder of the infection period. This level was maintained, despite diminishing numbers of worms, due to the growth of surviving individuals to a weight exceeding the original weight at maturity by a factor of more than 2. Experiments using rats that were mature at the time of infection demonstrated that the same response occurred, but approximately 3 weeks earlier.     

Effects of a 100 metacercarial cyst inoculum on the host-parasite relationship of Echinostoma caproni and ICR mice

The host-parasite relationship of a 100 metacercarial cyst inoculum of Echinostoma caproni in the ICR mouse was examined. Three groups of mice, A, B and C, each with six mice per group were used and all mice were necropsied at 14 days postinfection (p.i.), at which time the worms were ovigerous. Group A consisted of uninfected controls, whereas group B received 25 cysts per mouse (low dose) and group C received 100 cysts per mouse (high dose). There was no significant difference in food consumption between any of the groups from 0 to 14 days p.i. Control mice increased their body weight by 12%, group B by 5%, and group C showed a less than 1% increase in body weight between 0 and 14 days p.i. Echinostome parasitism caused a significant increase in the diameter of the mouse gut, with the gut of group C being more significantly dilated than that of either group A or B. The average worm recovery from group B was 20 worms per host, compared to 72 worms per host from group C. The mean wet and dry weights per worm from group B were 2.4 and 0.4 mg, respectively as compared to 0.6 and 0.2 mg respectively for group C. The mean number of uterine eggs per worm from group B was 180 compared to 125 for worms from group C. Worms from group C were more widely distributed in the small intestine than those from group B. Crowding effects associated with the high dose infection were clearly demonstrated in E. caproni from ICR mice.     

Single and multiple worm infections of Echinostoma caproni (Trematoda) in the golden hamster

Six of 10 hamsters fed a single metacercarial cyst of Echinostoma caproni (single-worm infections) and 13 of 19 hamsters fed either 2 or 5 cysts (multiple-worm infections) were infected with adult echinostomes at necropsy 22 days post-infection. Considerable histopathological changes to the small intestine occurred in hamsters carrying single-worm infections. There were no differences in either mean length, width or wet weight of echinostomes in single- versus multiple-worm infections. The mean number of eggs/worm from single-worm infections (525) was significantly greater than that from multiple-worm infections (288). The average percentage of fully developed miracidia/worm from single worms (94%) was similar to that from worms in multiple infections (92-95%). Single worms of E. caproni were capable of self-fertilization and production of viable eggs. Miracidia derived from single worms were as capable of infecting laboratory-reared Biomphalaria glabrata and producing patent rediae as were those from multiple infections.     

Mannose and the ‘Crowding effect’ of Hymenolepis in rats

Hymenolepis diminuta from rats infected with 10 cysticercoids and fed on a diet containing 3% (w/w) mannose for 4 weeks were found to be, on average, much heavier in terms of dry weight (46 mg) than those from rats fed on diets containing an equivalent concentration of either galactose, glucose or fructose (18 mg). Subsequently, the numbers, egg production and dry weights of worms were determined from rats which had been infected with doses varying from 0 to 160 cysticercoids per rat and fed on diets containing either 0, 1, 4, or 8% mannose (w/w). Density-dependent decreases in both worm dry weight and egg production were detected at 5 weeks post-infection, but both the number of worms recovered and their prepatent period appeared to be independent of cysticercoid dose. Differences in the distribution pattern of individual worm dry weights were observed between rats harbouring low (1–15) and high (16–135) worm burdens. Worms recovered from rats infected with 10 cysticercoids and fed on a diet containing 4% mannose (w/w) for 3 weeks were found to be, on average, more than twice as heavy (37 mg) as those from rats fed on diets containing 1, 2, or 8% mannose (w/w) for equivalent periods of time (16 mg). The results indicate that the ‘crowding effect’ cannot be explained simply in terms of inter-worm competition for carbohydrate.     

Density-dependent effects on the survival and growth of the rodent stomach worm Protospirura muricola in laboratory mice

The spirurid nematode, Protospirura muricola, is of intrinsic interest as a rodent model of gastric nematode infections. Since worm burdens can be very heavy in nature, density dependent processes may constrain parasite growth. Laboratory mice (BKW) were exposed to varying doses of infective larvae of P. muricola in the range 5 to 40 third-stage larvae (L3), in four separate experiments in which progressively higher doses were utilized. All mice were culled 60 days after infection and a total of 518 worms (226 male and 292 female worms) was recovered, measured and weighed. Overall survival was 58.9%, but survival declined significantly with increasing dose by approximately 21% (from 66% at 5 L3 per mouse to 52% at 40 L3 per mouse). The length and weight of worms correlated positively in both sexes. Total worm biomass increased linearly with increasing numbers of worms. However, whilst the length and weight of male worms declined with increasing worm burden (8.4 and 24.6% respectively), female worms were less affected, only length showing a significant reduction with increasing parasite burden (16.0%). Therefore, increasing worm burdens impeded growth of P. muricola, but reduction in length and weight were relatively small in relation to the overall size of this nematode. Increasing worm burdens were associated with loss of host weight and reduction in stomach weight and worm burdens in excess of 20 exerted a measurable cost to the host, which in the field, may be associated with loss of overall host fitness.     

Animal Models for Echinostoma malayanum Infection: Worm Recovery and Some Pathology

Echinostomes are intestinal trematodes that infect a wide range of vertebrate hosts, including humans, in their adult stage and also parasitize numerous invertebrate and cold-blooded vertebrate hosts in their larval stages. The purpose of this study was to compare Echinostoma malayanum parasite growth, including worm recovery, body size of adult worms, eggs per worm, eggs per gram of feces, and pathological changes in the small intestine of experimental animals. In this study, 6-8-week-old male hamsters, rats, mice, and gerbils were infected with echinostome metacercariae and then sacrificed at day 60 post-infection. The small intestine and feces of each infected animal were collected and then processed for analysis. The results showed that worm recovery, eggs per worm, and eggs per gram of feces from all infected hamsters were higher compared with infected rats and mice. However, in infected gerbils, no parasites were observed in the small intestine, and there were no parasite eggs in the feces. The volume of eggs per gram of feces and eggs per worm were related to parasite size. The results of histopathological changes in the small intestine of infected groups showed abnormal villi and goblet cells, as evidenced by short villi and an increase in the number and size of goblet cells compared with the normal control group.     

Influence of host strain and helminth isolate on the first phase of the relationship between rats and Moniliformis moniliformis (Acanthocephala)

Eleven trials, involving 440 rats bred from 3 laboratory strains and worms from 4 isolates of Moniliformis moniliformis, were carried out with each rat receiving an oral dose of 15 cystacanths. The results showed that the infectivity of the cystacanths was not affected by their age (range 55-194 days) or by their density per cockroach during development (16.1-88.6 cystacanths per cockroach). The numbers of worms per rat recovered at 35 days postinfection (p.i.) were shown to be related to rat strain, with highly inbred strains (PVG and F344) being more supportive of numbers of worms than an outbred Wistar strain. There was no evidence to suggest that the sex of the rats had any influence on the numbers of worms recovered at 35 days p.i. Evidence was obtained to suggest that smaller (younger) rats are likely to support more worms on average than larger (older) rats. There was no evidence of any relationship between worm weight and numbers of worms present per rat on day 35 p.i. Generally, rat strain had little effect on the dry weight (growth) of male M. moniliformis, in contrast to observations made for female worms. The greatest range of worm weights was observed from the recent isolate of the worm (1982) as compared with the well established isolate (1956) and the rats that supported most worms differed from those that harbored the largest worms. Rat sex was not observed to be associated with worm weight. The frequency distributions of numbers of M. moniliformis per rat were not described readily by the negative binomial distribution.     

Possible immunological damage to the tegument of Hymenolepis diminuta in mice and rats.

Opaque or darkened areas (DA) of variable size and position occur on Hymenolepis diminuta in mice and rats. In mice DA normally first appear in the neck region of the worm but subsequently they appear elsewhere and increase in number until destrobilation or worm expulsion. The posterior of destrobilated worms is often darkened. In the more immunogenic infections with six cysticercoids there are more DA per worm than in infections with one cysticercoid. DA are areas of the tegument with a homogeneous increase in electron density; abnormal mitochondria; reduced granular endoplasmic reticulum, Golgi complexes and discoidal secretory bodies; and accumulation of lipid droplets. DA disappear from worms maintained for up to 4 h in Hanks' balanced salt solution and can be induced by mechanical damage to the worms. As the numbers of DA increase with the duration and intensity of infection and have similarities with types of cell injury, they are probably sites of worm pathology induced by host immunity.     

Infections of forest rat filaria, Breinlia booliati, in neonate and juvenile laboratory white rats

The kinetics of Breinlia booliati infection in 3 inbred rat strains (Lewis, Wistar and Sprague Dawley) were investigated. One group of rats was infected as neonates (less than 24 hours of age) with third-stage larvae of B. booliati and the other group was infected as juveniles (4 weeks of age). The results showed that infection in the neonates were significantly different from the infection in the juveniles. The 60 rats infected as neonates, when necropsied between 8 to 10 months postinfection, yielded adult worms. The 2 neonatal infection groups of Lewis and Wistar strains showed highest susceptibility to the infections. The mean prepatent period was 85 days. Ninety to 95% of the infected rats were patent with microfilaraemia and a large percentage (33 to 47%) of them had high microfilaraemia counts exceeding 3000 mff/20 mm3 of blood and larger sizes (mean 157.11 mm for female adult worms and 61.88 mm for male adult worms. The adult worms were distributed equally in both the pleural (57%) and peritoneal cavity (43%). In most aspects, the neonatal infection group of the Sprague-Dawley strain was intermediate in susceptibility between the 2 neonatal infection groups of the Lewis and Wistar strains and the 3 juvenile infection groups. In contrast to neonatal infection groups, the 3 juvenile infection groups exhibited low infection rates (37%, 58% and 47% for the Lewis, Wistar and Sprague Dawley strains respectively), longer prepatent periods (mean 101 days), lower recovery rates (2 to 4%), lower adult worm loads (mean 0.4 to 0.8 female worms, and 0.2 to 0.8 male worms per rat), and smaller sizes (mean 141.24 mm for female adult worms and 53.75 mm for male adult worms). Forty-four to 57% of these infected rats harboured either single male or single female adult worms in the body cavity. Most (92%) of the adult worms recovered from the juvenile infection groups resided in the pleural cavity and the remaining 8% were recovered from the peritoneal cavity. Microfilaraemia could be detected in only 3/20 Lewis rats, 5/20 Wistar rats and 5/20 Sprague Dawley rats. The mean peak microfilaraemia of the 3 pooled juvenile infection groups was 632 mff/20 mm3 of blood, ranging from 7 mff/20 mm3 to 1856 mmf/20 mm3. Our results indicate that the susceptibility to B. booliati infection in white rats is both genetic and age-associated. The responses of the 2 distinct infection groups to B. booliati infections are discussed.     

Immunological aspects in the rejection process of Hymenolepis muris-sylvaticae from CFLP and NMRI mice

When mice were treated with 1.25 mg cortisone acetate thrice weekly, recovery of Hymenolepis muris-sylvaticae was significantly higher than in untreated controls, both in oral infections with six cysticercoids and surgical transplantations of one 7-day or 8-day-old worm. Cortisone treatment also resulted in the worms being located more anteriorly in the small intestine. Evidence of an immunological response against the tapeworm in the intestine is given by: an accelerated rejection of a secondary oral cysticercoid infection and a significant difference of the dry weights of the worms recovered on day 10 in CFLP mice; an accelerated rejection of a secondary surgical infection on days 4 and 6 in CFLP mice and on days 3 and 4 in NMRI mice; an accelerated rejection of a secondary surgical infection given 3 and 6 months after the primary immunizing infection in SWISS-albino mice.     

Cortisone-sensitive, innate resistance to Hymenolepis nana infection in congenitally athymic nude rats

The innate resistance of the unnatural rat host to the mouse tapeworm Hymenolepis nana is cortisone sensitive but thymus independent. When congenitally athymic nude rats were orally given eggs, cysticercoids, or adult worms of H. nana, no lumenal adults were established except when they were treated with cortisone acetate during the expected lumenal development. The effect of cortisone to promote adult maturation in the rats was compared in nude and normal rats given eggs of H. nana. The fecundity of the worms (assessed by the fresh worm biomass and the number of infective eggs produced) was much higher in cortisone-treated nude rats than in cortisone-treated normal rats. When the nude rats reconstituted with thymocytes were given eggs and treated with cortisone, the fecundity of H. nana dropped to the same level as in cortisone-treated normal rats. It is strongly suggested that the unnatural rat host has thymus-independent cortisone sensitive resistance to an initial infection (which is the main component of the innate resistance and blocks the lumenal establishment of this parasite) and thymus-dependent resistance (which suppresses the established worms' fecundity and may be ascribed to acquired resistance to the ongoing infection).     

Factors influencing the fecundity of Moniliformis moniliformis (Acanthocephala): constant diet and varied dose

Aspects of the reproductive performance of Moniliformis moniliformis were investigated in rats allowed to feed ad libitum on a purified diet containing 1% (w/w) fructose as an energy source for the worms. The rats were infected with either 10, 20, 40 or 80 cystacanths each with the intention of investigating density-dependent effects on worm fecundity. The establishment of the worms in the gut was independent of dose, but survival, growth and reproductive performance generally were shown to be related to the infective dose given to the rats. The effects could not be related to the absolute numbers of worms present in the small intestine at post-mortem examination. In general, some unidentified regulatory process appeared to operate to create severe density-dependence in survival so that surviving parasites were not present in numbers expected to generate competition. Attainment of sexual maturity, growth and the production of mature eggs by worms from rats given doses of 80 cystacanths each were delayed compared with worms from rats given the other doses, but eventually the performance of the high-dose worms caught up. Worms attached more anteriorly in the small intestine grew bigger and produced more mature eggs. Possible mechanisms responsible for the observed effects are discussed.     

Efficacy of ivermectin in hookworms as examined in Ancylostoma caninum infections

Single oral doses of ivermectin were given to dogs with moderate or heavy infections of Ancylostoma caninum (egg counts ranging from 7,100 to 41,700 eggs/g feces) at 100, 50, 30, or 10 micrograms/kg body weight. Each of these dosages was effective in clearing the infection completely, so that numerous worms were passed in the feces on days 1-3, but no worm was recovered from the intestinal tract at necropsy on day 4 after treatment. In contrast, an average of 178 worms per dog was recovered at necropsy from the vehicle-treated control and the untreated animals. Albendazole, a known anti-hookworm agent, even in a dose of 400 mg, eliminated only 21-65% of the worms harbored by the infected animals. No untoward reaction to ivermectin or significant pathological change was noted in the experimental animals. In vitro experiments demonstrated that ivermectin: (1) was highly detrimental to actively motile adult worms in concentrations greater than 5.60 micrograms/ml; (2) was detrimental to eggs inside the uterine tissue of female worms in dosages at or greater than 10 micrograms/kg body weight; and (3) killed infected larvae in concentrations as low as 0.0025 micrograms/ml.     

Reproductive potential of Echinococcus multilocularis in experimentally infected foxes, dogs, raccoon dogs and cats

A total of 15 red foxes, 15 raccoon dogs, 15 domestic dogs and 15 domestic cats were each infected with 20,000 protoscolices of Echinococcus multilocularis. At 35, 63, and 90 days post inoculation (dpi), five animals from each group were necropsied and the worm burdens determined. The highest worm burdens in foxes (mean of 16,792) and raccoon dogs (mean of 7930) were found at 35 dpi. These declined to a mean of just 331 worms in foxes and 3213 worms in raccoon dogs by day 63 with a further decline to 134 worms in foxes and 67 worms in raccoon dogs by day 90. In dogs, there was no significant difference between worm burdens recovered at days 35 (mean of 2466) and day 90 (mean of 1563), although reduced numbers were recovered on day 63 (mean of 899). In cats, worms were found in four animals 35 dpi (mean of 642), in three at 63 dpi (mean of 28) and in two at 90 dpi (mean of 57). Faecal egg counts were determined at 3 day intervals from 25 dpi. A mathematical model of egg excretion dynamics suggested that the mean biotic potential per infected animal was high in foxes (346,473 eggs); raccoon dogs (335,361 eggs) and dogs (279,910 eggs) but very low for cats (573 eggs). It also indicated that approximately 114, 42 and 27 eggs per worm were excreted in the faeces of dogs, raccoon dogs and foxes, respectively. The fecundity of worms in cats was low with an average of less than one egg per worm. The peak levels of coproantigen were detected earlier in foxes and raccoon dogs than in dogs. Eggs recovered from foxes, raccoon dogs and dogs resulted in massive infections in experimental mice. However, metacestodes did not develop from eggs originating from infected cats. It is concluded that foxes, raccoon dogs and dogs are good hosts of E. multilocularis. In contrast, the low worm establishment, the very few excreted eggs and the lack of infectivity of eggs strongly indicate that cats play an insignificant role in parasite transmission.     

Hymenolepis nana: worm recovery from congenitally athymic nude and phenotypically normal rats and mice

When eggs or mouse-derived cysticercoids of Hymenolepis nana were inoculated into previously uninfected congenitally athymic nude (rnu/rnu) rats of an outbred Rowett strain, they failed to mature in the intestinal lumen. They also failed to mature in phenotypically normal (rnu/+) littermates, except when these hosts were treated with cortisone acetate from the beginning of the lumen phase. The Rowett rat, either thymus-deficient or not, was susceptible to tissue cysticercoids but resistant to luminal adults. It is therefore considered to be an unnatural host, at least for mouse-derived H. nana. There was little or no difference in susceptibility to initial tissue cysticercoids between these nude rats and phenotypically normal ones. The normal rats became completely resistant to reinfection with eggs and no secondary cysticercoids developed in their intestinal tissue, whereas the nude rats showed unaltered susceptibility to secondary tissue cysticercoids. Thus, acquired resistance to egg challenge, assessed by the failure of tissue cysticercoid recovery, was thymus-dependent. However, innate resistance to both a primary egg dose, assessed by the low recovery rates of tissue cysticercoids, and to a primary cysticercoid dose, assessed by the failure of luminal adult recovery, were thymus-independent. The effect of cortisone acetate to initiate maturation of H. nana appeared to be unrelated to thymus function. In contrast, all mice, either thymus-deficient or not, were highly susceptible to both phases. The number of worms recovered was more than 10 times greater than that of cysticercoids established in the rat's intestinal tissue.(ABSTRACT TRUNCATED AT 250 WORDS)     

Recovery,growth, and development of Acanthoparyphium tyosenense (Digenea: Echinostomatidae) in experimental chicks

Chicks were experimentally infected with Acanthoparyphium tyosenense (Digenea: Echinostomatidae) metacercariae per os, and the growth and development of worms in this host were observed from days I to 38 postinfection (PI). The worms grew rapidly and matured sexually in the small intestine (chiefly in the jejunum) of chicks by day 5 PI. and survived at least up to day 38 Pi, although worm recovery decreased after day 5 PI. Both parenchymal and reproductive organs increased greatly in size from day 2 to day 10 PI and then continued to increase gradually in size up to day 38 PI. The number of uterine eggs reached a peak on days 10 and 15 PI and then decreased gradually. The results suggest that chicks are a fairly suitable definitive host for experimental infection with A. tyosenense.     

Hymenolepis citelli and H. diminuta: worm burdens in homologous and heterologous infections in rats

Primary and secondary infections of Hymenolepis citelli were followed in CFHB rats. In primary infections of 6, 20, and 50 cysticercoids, over 75% of the worms administered became established and grew. Thereafter, survival depended on the intensity of the primary infection. Acquired resistance to homologous challenge infections could be stimulated in rats by prior infection with H. citelli. The growth of secondary worms decreased as the intensity of the sensitizing infection increased. The protective response waned with time in the absence of the primary worms. Cross-protective responses between H. citelli and H. diminuta occur in rats infected previously with either parasite. Retardation in the growth of secondary worms may have an immunological basis.     

Establishment of adult Parelaphostrongylus tenuis, patent infections, and acquired immunity after experimental infection of white-tailed deer (Odocoileus virginianus) and red deer (Cervus elaphus elaphus)

Experimental Parelaphostrongylus tenuis infections were established in white-tailed deer (Odocoileus virginianus) and an atypical host, red deer (Cervus elaphus elaphus). Groups of deer were fed 10, 25, or 100 third-stage larvae (L3) of P. tenuis and received a single equivalent challenge exposure at varying intervals. Infections were monitored up to 6 yr in white-tailed deer and up to 2.8 yr in red deer. The prepatent period in white-tailed deer varied from 91 to 1,072 days (381 +/- 374) and in red deer from 105 to 358 days (167 +/- 77). Adult worms lived for up to 6 yr in white-tailed deer. Although most had patent infections until necropsy, latent periods were observed regardless of season. Adult worms lived for up to 2.8 yr in red deer, and patent infections persisted for 20-363 days (152 +/- 106). Patent infections were correlated with the presence of adult worms in blood vessels and sinuses of both deer species. Worms were restricted to the subdural space in all deer with latent and occult infections. Adult worm recovery in white-tailed deer fed 10 or 25 L3 corresponded to the mean intensities reported in natural infections of white-tailed deer Recovery from deer fed 100 L3 was not typical of natural infection intensities. Adult P. tenuis established in all groups of red deer, but neurologic disease was restricted to animals fed 100 L3. Acute neurologic disease was associated with subdural hemorrhage and occurred at 11 mo postinfection in 2 red deer. The absence of postchallenge patent periods and the persistence of occult infections indicated that challenge exposures did not establish. These data indicate that acquired immunity to P. tenuis was established by 6 mo postinfection in both white-tailed and red deer. Latent periods in white-tailed deer and latent infections in red deer reinforce the need for a reliable diagnostic assay.     

Brugia pahangi in rats: increasing the number of infective larvae inoculated increases the percentage of microfilaremic rats

One hundred Brugia pahangi infective larvae (L3) caused microfilaremic (mf + ve) infection in 56% of inbred PVG rats. Adult worms were recovered consistently from infected rats but worm recovery was very low, only 1-3% of L3 inoculated survived to adulthood and the worms were dispersed in a wide range of anatomical sites. This suggested that lack of microfilaremia may be due to the low probability of male and female worms meeting in the same site and thus may be numerically and topographically based. When the number of infective larvae inoculated was increased to 500, the percentage of mf + ve infections in rats also increased to 94%, corroborating the hypothesis that lack of mf was not due to an immune response. In a further experiment all infected rats had lost both mf and adult worms by day 420. It has yet to be established whether final rejection of the parasite is due to immunity.     

Contamination of fox hair with eggs of Toxocara canis

Eighty-seven red foxes were investigated for the presence of Toxocara eggs on a sample of their hair from the peri-anal region. The worm burden of Toxocara in each fox intestine was also assessed and the relationship between eggs on the hair and worms in the intestine explored. Twenty-eight per cent of the foxes were found to have Toxocara eggs on their hair, with an average of 1.31 +/- 3.21 eggs per gram of hair (+/- SD). Sixty-one per cent of foxes harboured Toxocara worms within their intestines, with a mean worm burden (+/- SD) of 4 +/- 8. Host age and sex did not significantly influence the observed eggs on the hair or the worm burden. No significant correlation was found between the numbers of eggs on the hair and the worm burden within the intestine. These data collected from foxes are explored in the context of data from stray dogs and the possible epidemiological differences are discussed.