Changes in tegumental surface during development of Schistosoma mansoni

The tegumental surface of immature Schistosoma mansoni was studied with the scanning electron microscope. The surfaces of immature males and females bear no resemblance to that of adult worms and are characterized by having many tegumental folds. The tegumental surfaces of immature males and females are similar, and the dorsal and ventral surfaces of the male are similar before formation of the gynecophoral canal. Transition of the tegumental surface from the juvenile to the adult form begins after worms are in copula and have grown to several millimeters in length.     

Hybridisation between the digeneans Schistosoma haematobium and S. mattheei : viability of hybrids and their development in sheep

The F1 and F2 hybrids of Schistosoma haematobium male × S. mattheei female were studied with regard to infectivity to intermediate and definitive hosts, isoenzymes (phosphoglucomutase) of individual male worms, randomly amplified polymorphic DNAs of individual adult worms and scanning electron micrographs of the tubercles of male worms. The infection rate of the F1 hybrid miracidia in Bulinus globosus (41.7%) was greater than that achieved in B. wrighti (16.3%); the infection rate of the F2 in B. wrighti was 15.4%. In the definitive hosts: in sheep only male F1 hybrids (i.e. no females and no F2 worms)were recovered; but in hamsters both paired F1 worms and unpaired F1 males were recovered, as were one pair of worms and unpaired males of the F2 generation. The S. mattheei and S. haematobium male worms showed very distinctive PGM patterns, and the F1 hybrids showed additive patterns and a polymorphism with two distinct types of band patterns which are the result of polymorphism in the S. haematobium. The RAPD profiles of the F1 hybrids were also composite of the two parental species. Scanning electron micrographs of the tubercles of male S. haematobium showed them to be heavily spined, whereas those of S. mattheei males were devoid of spines. The F1 hybrids did show variation ranging from non-spined, some with partial spination, to those with heavily spined tubercles. Male worms of the F2 generation possessed tubercles either with or without spines. The potential significance of hybridisation in areas of sympatry between S. haematobium and S. mattheei is discussed.     

Gastrointestinal myiasis by larvae of Sarcophaga sp. and Oestrus sp. in Egypt: report of cases, and endoscopical and morphological studies

Four cases of gastric or intestinal myiasis are reported. The cases contain 2 males (1 child 10 years old, and 1 adult 40 years old) and 2 females (1 girl 18 years old, and 1 adult 50 years old) from Minia Governorate, Southern Egypt. Three of them, including cases no. 1, 3, and 4, were gastric myiasis, and complained of offensive hematemesis of bright red blood. Minute moving worms, larvae of the fly, were found in the vomitus. On the other hand, case no. 2 had intestinal myiasis, and complained of abdominal distention, nausea, vomiting, and diarrhea. The stool of case 2 was mixed with blood, and minute moving worms were observed in the stool. Endoscopy was performed to explore any pathological changes in the stomach of the patients. The larvae were collected and studied macroscopically, microscopically, and us-ing a scanning electron microscope (SEM) to identify their species. Three different types of larvae were identified. The larvae isolated from case 1 were diagnosed as the second stage larvae of Sarcophaga species, and the larvae isolated from case 2 were the third stage larvae of Sarcophaga species. On the other hand, the larvae isolated from cases 3 and 4 were diagnosed as the third stage larvae of Oestrus species.     

Transmission electron microscopic observations on lipid release in Schistosoma mansoni maintained in vitro

Transmission electron microscopy was done on Schistosoma mansoni adult males and females unpaired immediately following recovery from mice and on worms maintained in Earle's balanced salt solution (EBSS) containing 0.1% glucose and 0.5% lactalbumin hydrolysate for 1.0 h at 37°C. Males incubated for 0.5 and 1.0 h released lipids from non-tuberculate and tuberculate areas of the dorsal tegument. Lipid release was not seen in females. Transmission electron microscopy revealed no apparent damage in worms maintained in BBSS for 1.0 h. Our observations suggest that structures described as “spheres” (Weisberg, Carlisle &; Bentley, 1983) are lipid droplets.     

Location,morphology and histology of sex pheromone glands of the female gypsy moth, Lymantria dispar (L.)

Scanning electron microscopy, histology and a male wing fanning bioassay were used in this study to locate the sex pheromone-producing glands of the female gypsy moth, Lymantria dispar. When exposed to female sex pheromone, adult males exhibit a strong wing fanning behaviour prior to take off. We found that adult males showed positive response to calling females and to tissue extract from both dorsal and ventral portions of the intersegmental membrane between 8th and 9th-abdominal segments. A typical male response usually starts with elevation of antennae, movement of head in different directions, walking, wing fanning and onset of search flight. Histological and scanning electron microscopic studies suggested that the sex pheromone glands are located on the dorsal and ventral aspects of the intersegmental membrane. The glands appear as two highly convoluted integumentary areas with hypertrophied glandular epidermal cells.     

Bone-eating Osedax females and their 'harems' of dwarf males are recruited from a common larval pool

Extreme male dwarfism occurs in Osedax (Annelida: Siboglinidae), marine worms with sessile females that bore into submerged bones. Osedax are hypothesized to use environmental sex determination, in which undifferentiated larvae that settle on bones develop as females, and subsequent larvae that settle on females transform into dwarf males. This study addresses several hypotheses regarding possible recruitment sources for the males: (i) common larval pool--males and females are sampled from a common pool of larvae; (ii) neighbourhood--males are supplied by a limited number of neighbouring females; and (iii) arrhenotoky--males are primarily the sons of host females. Osedax rubiplumus were sampled from submerged whalebones located at 1820-m and 2893-m depths in Monterey Bay, California. Immature females typically did not host males, but mature females maintained male 'harems' that grew exponentially in the number of males as female size increased. Allozyme analysis of the females revealed binomial proportions of nuclear genotypes, an indication of random sexual mating. Analysis of mitochondrial DNA sequences from the male harems and their host females allowed us to reject the arrhenotoky and neighbourhood hypotheses for male recruitment. No significant partitioning of mitochondrial diversity existed between the male and female sexes, or between subsamples of worms collected at different depths or during different years (2002-2007). Mitochondrial sequence diversity was very high in these worms, suggesting that as many as 10(6) females contributed to a common larval pool from which the two sexes were randomly drawn.     

Asymmetric learning to avoid heterospecific males in Mesocricetus hamsters

If a female mates with a male of a closely related species, her fitness is likely to decline. Consequently, females may develop behavioral mechanisms to avoid mating with heterospecific males. In some species, one such mechanism is for adult females to learn to discriminate against heterospecific males after exposure to such males. We have previously shown that adult, female Syrian hamsters (Mesocricetus auratus) learn to discriminate against male Turkish hamsters (Mesocricetus brandti) after exposure to a single heterospecific male during 8 days across a wire-mesh barrier. Here we repeated that experiment but this time we exposed female Turkish hamsters to a male Syrian hamster for 8 days and then measured sexual and aggressive behaviors towards that heterospecific male and towards a conspecific male. In contrast to female Syrian hamsters, female Turkish hamsters did not differ in their latency to go into lordosis or in any measure of aggression towards either type of male. Female Turkish hamsters spent less time in lordosis with the heterospecific male, but the percentage of trials in which females copulated with conspecific and heterospecific males did not differ. When comparing females from both species that had been exposed to a heterospecific male for 8days, female Syrian hamsters copulated less and were more aggressive towards the heterospecific male compared to the behavior of female Turkish hamsters. We discuss how this asymmetric response between females of the two species may be due to the much larger geographical range of Turkish hamsters compared to Syrian hamsters.     

Tegumental changes in adult Schistosoma mansoni harbored in mice treated with artemether

A detailed temporal examination was made of alterations induced by artemether in the tegument of adult Schistosoma mansoni worms using scanning electron microscopy (SEM). Mice infected with S. mansoni cercariae 42 days previously were treated intragastrically with artemether at a single dose of 400 mg/kg. Groups of 3 mice were killed at 24 hr, 72 hr, and 7 days after treatment; the worms were collected by perfusion and examined by SEM. Twenty-four hours after artemether treatment, focal damage to the tubercles on the tegumental surface of male worms was seen. In both male and female worms, there was focal swelling and fusion of tegumental ridges, and sometimes peeling. After 72 hr, the damage to the tegument had increased, especially in female worms, with extensive swelling, fusion, and peeling of the tegumental ridges. In the most severely damaged worms, host leukocytes were seen to be adhered to the damaged tegument. Damage to the oral sucker was also occasionally seen in both male and female worms. Seven days after treatment, the appearance of the tegument had returned to normal in some male and female worms, whereas others still showed apparent damage. The results demonstrate that artemether damages the tegument of adult S. mansoni, and the intensity of damage is more severe in female worms than in males.     

Changes in behaviour associated with pair formation in the polychaete Harmothoë imbricata (L.)

Spacing between individuals in populations of Harmothoë imbricata has been investigated both on the shore and in the laboratory. Males tend to occur closer together than females, and the mean male‐male individual distance measured was less than the mean distance between females; male—female distances for immature worms were intermediate. When worms mature they pair: the male mounts the female and lies across her dorsal surface. There is evidence that after spawning the members of a pair separate. Contact responses between worms have been investigated in the laboratory. Most encounters between immature worms lead to separation, as a result of one or both worms moving rapidly away from the other or fighting; females show more marked avoidance behaviour than males. The majority of male‐male and female‐female encounters between mature worms also lead to separation but in male‐female encounters the male usually mounts the female. A male which has mounted a female becomes highly aggressive and will attack intruding males but not females.     

Comparison of surface topography of three species of Diphyllobothrium (Cestoda,pseudophyllidea) by scanning electron microscopy

Plerocercoids of different sizes as well as adult worms of D. dendriticum, D. latum and D. ditremum were studied using scanning electron microscopy (S.E.M.). In the plerocercoids there were found distinct differences in appearance and length of microtriches between these three species, while the microtriches of adult worms were more similar. A regional difference in microtrix appearance was found in the larvae of D. ditremum and D. dendriticum. This was not apparent with S.E.M. in adult worms. The length of ‘body’ microtriches in D. dendriticum varied with the length of the larvae. The topography of the genital atrium of mature and gravid proglottids in adult worms of these three species is also described.     

Investigation of venereal, transplacental, and contact transmission of the Lyme disease spirochete, Borrelia burgdorferi, in Syrian hamsters.

A hamster was inoculated with the SI-1 strain of Borrelia burgdorferi and subsequently served as a host to larval Ixodes scapularis Say. Approximately 68% of the nymphs resulting from the fed larvae were infected. Nymphs from this group were fed on uninfected hamsters, and 3 of 4 males and 6 of 6 females became infected. The infected hamsters were allowed to mate with uninfected partners to test for venereal transmission. Six infected females were mated with 6 uninfected males, whereas 3 infected males were mated with 6 uninfected females. None of the uninfected hamsters became infected after mating. Two protocols were used to determine if transplacental transmission of B. burgdorferi occurred. One group included 6 nonpregnant infected females that were subsequently mated and became pregnant. Three of the females were allowed to carry to full term, whereas the other 3 were killed prior to parturition. All fetuses and offspring were negative for B. burgdorferi based on cultures and monoclonal antibody assays. Another group of 6 females was infected via tick bite after becoming pregnant; those females were allowed to carry fetuses to birth and all were negative. Attempts at contact transmission of B. burgdorferi from 2 infected females to 2 uninfected male and 2 uninfected female hamsters and from 2 infected males to 2 uninfected male and uninfected female hamsters via urine or feces failed.     

Effects of low-protein diet on Schistosoma mansoni morphology visualized by morphometry and confocal laser scanning microscopy

Previous studies have shown that protein deficiencies can hamper both the course of experimental schistosomiasis and normal development of adult worms. To further investigate this relationship, we compared adult male and female Schistosoma mansoni from malnourished and well-fed mice through morphometric and confocal laser scanning microscopy analysis. Swiss mice were fed protein-deficient diets (8%) and infected subcutaneously with approximately 80 S. mansoni cercariae (BH strain, Brazil). Control mice were fed a standard rodent diet (23% protein). The nutritional status was evaluated by body weight gain and albumin values. Mice were sacrificed 63 days post-infection. Recovered worms were stained with hydrochloric carmine and preserved as whole-mounts for bright-field examination and confocal microscopy. The body weight gain and serum albumin concentrations were significantly lower (P < 0.05) in malnourished mice than in controls. In general, all morphometric values of specimens grown in malnourished mice were lower than those of control mice. Schistosome worms grown in malnourished mice had statistically significant differences (P < 0.05) in the reproductive system and tegument than those grown in mice fed standard diets. In female worms, vitelline glands showed few remaining follicles and ovaries lacked mature oocytes. In male parasites, tubercles were fewer in number on the dorsal surface and testicular lobes presented fewer differentiated germinal cells. In summary, we describe novel data supporting the view that low-protein diets may influence the development of adult worms.     

Development of male-incubated ovaries in the gypsy moth,Lymantria dispar

Ovaries from Lymantria dispar females were transplanted into an environment lacking vitellogenin, the male milieu, in order to determine how the presence of vitellogenin in the hemolymph affects the process of protein uptake by gypsy moth oocytes. When undeveloped ovaries from newly ecdysed last instar females were transplanted into males of the same stage, follicles detached from the germarium and increased in size, but the growth of oocytes proceeded more slowly than those from female controls. Although chorion fromation was delayed in male-grown ovaries, scanning electron microscopy of chorionated eggs recovered from adult males showed that a chorion with normal surface architecture was formed by the adult stage. SDS-PAGE analysis of the male-grown ovaries and hemolymph from males receiving ovaries showed that vitellogenin production was not stimulated by the organ transplant and only male hemolymph proteins were internalized by the male-incubated ovaries. Thus, in the absence of vitellogenin, endocytosis of male hemolymph proteins occurred, but the rate of oocyte growth was slowed.     

Dipetalonema viteae: primary, secondary and tertiary infections in hamsters.

Hamsters were given primary infections of 100, 200, and 300 D. viteae larvae and groups killed at various intervals after infection. In addition, hamsters were sequentially infected with 100, 200, and 300 larvae and groups killed at 100 or 75 days after the secondary and tertiary infection, respectively. Blood microfilariae were detected on Day 60 following a primary infection, reached a maximum on Day 75, declined to low levels by Day 105, and were negative on Day 120. No microfilariae reappeared in the blood of hamsters given secondary or tertiary infections.Between 20–30% of the infecting larval dose had reached the adult stage by Days 75 or 100 postinfection in hamsters given primary, secondary, or tertiary infections. There was no evidence of arrested larval development in hamsters receiving a second or third challenge infection. Almost half of the tertiary infection hamsters developed subcutaneous nodules and their numbers varied greatly among individual animals. The nodules variously contained living worms, pus, and fragmented worms, or pus only. Hamsters given primary infections of 100, 200, or 300 larvae and killed 375 days after infection had no subcutaneous nodules; however, hamsters given the 200 and 300 larval infections were seen to have dead worms in the subcutaneous tissues. No stunting of adult worms was noted and all female worms had uteri packed with microfilariae.     

The Infestiveness of Heat and Gold Exposed Diphyllobothrium Latum Plerocercoids on Golden Hamster

The infestiveness of heat (56°C/5 min. and 50°C/5 min.), and cold (−10°C and −6°C) exposed Diphyllobothrium latum plerocercoids was studied by administration to golden hamsters (Mesocricetus auratus). The cold exposed larvae were subjected to an exposure, analogous the freezing of fish and which is cut off when the temperature has declined either to −6°C or −10°C. The plerocercoids were administered to the hamsters under inhalation anaesthesia, 5 larvae per hamster. Out of 150 normal control plerocercoids administered on 30 hamsters, 74 adult worms developed, indicating 49 % infestiveness. Tests carried out with 105 56°C/5 min. exposed larvae on 21 hamsters, 45 50°C/5 min. exposed larvae on 9 hamsters, and 100 larvae exposed to −10°C on 20 hamsters all gave negative results, while out of 50 −6°C exposed larvae on 10 hamsters 3 developed to adult worms. The results show that the 56°C/5 min. and −10°C exposed larvae, which in previous studies have been considered to be inactivated on the basis of the resultant permanent immobilization, were not able to develop further in the host. Thus permanent immobility seems to be a reliable criterion of larval death.     

Re-establishing a life cycle of Schistosoma mansoni from cryopreserved larvae

To study the feasibility of re-establishing a life cycle of Schistosoma mansoni (NMRI strain) from cryopreserved larvae, schistosomules were suspended in the cryoprotectant 1,2-ethanediol and cryopreserved in liquid nitrogen. Mice were injected intramuscularly with samples thawed after 3 days, 3 wk, or 6 mo in liquid nitrogen storage. Two to 5% of the cryopreserved larvae and approximately 18% of corresponding unfrozen control larvae developed into adult worms. Infectivity did not decrease as a function of storage time. The adult worms showed no structural damage or changes in overall size and morphology when examined by light and transmission electron microscopy. Female worms derived from cryopreserved larvae had the same or slightly elevated egg production as controls, but tissue egg distributions were comparable. Subsequent passages through Biomphalaria glabrata snails and mice revealed no difference in snail prepatent death rate, percentage of snails infected, cercarial production per snail, or cercarial infectivity.     

Development of Heterodera glycines Ichinohe on Soybean,Glycine max (L.) Merr., under Gnotobiotic Conditions

The life cycle of the soybean cyst nematode, Race 3 (SCN 3), Heterodera glycines Inchinohe was determined from observations of the developmental stages on soybean Glycine max cv. Kent root explants under gnotobiotic conditions at 25 C. Approximately 51% of the second-stage larvae penetrated the root l day after inoculation (DAI). Third-stage larvae appeared 4 DAI, became sexually differentiated 5 DAI, and protruded from the root tissues 6 DAI. Fourth-stage males and females were observed 7 DAI. Ensheathcd adult males were observed at 9 DAI and exsheathed to free adults at 11 DAI. The fourth-stage female became an adult at l0 DAI, Males entwined arotmd the gelatinons sac of the female at 12 DAI and were assumed to be mating. Some males actually penetrated and were enveloped by the gelatinous sac. The female-to-male sex ratio ranged from 2.3 to 9.5:1. First- and second-stage larvae were observed in the egg 17 and 19 DAI, respectively. The life cycle of the SCN 3 was completed 21 DAI upon hatching of the eggs and emergence of second-stage larvae. The average number of eggs in the cyst body and gelatinous sac, was 210 and 187, respectively. Key words: reproduction, soybean cyst nematode, scanning electron microscopy.     

Effect of niclosamide on the tegumental surface of Haplorchis taichui using scanning electron microscopy

The effect of niclosamide on the tegument of adult Haplorchis taichui (Trematoda: Heterophyidae) exposed in vitro was observed by scanning electron microscope. Adult worms were incubated in Tyrode's solution containing 0.01, 0.1, 1.0, and 10 microg ml(-1) of niclosamide for 30 min, 1, 6, 12 and 24 h. Control groups were incubated in Tyrode's solution without niclosamide and worms remained active until 24 h. In 0.01 microg ml(-1) of niclosamide, worms showed slightly active movements up to 1 h after incubation, while in 0.1 microg ml(-1) solution a few worms showed only slightly active movements after 30 min. Tegumental changes were determined by scanning electron microscopy. Swelling and blebbing of the tegument were observed on both ventral and dorsal sides. After longer periods, extensive swelling and blebbing of the tegument became more severe and there was a loss of the apical plasma membrane in some regions. Empty spine sockets occurred, and small perforations penetrated the basal lamina, followed by some lesions. Destruction of both surfaces was more pronounced on the posterior compared with the anterior regions.     

Dracunculus insignis: experimental infection in the ferret, Mustela putorius furo

The laboratory study of dracunculiasis has suffered from the lack of a suitable, readily available animal model. We have been able to experimentally infect ferrets. Mustela putorius furo, with the North American dracunculid, Dracunculus insignis. Ferrets were inoculated with 75 to 100 infective larvae and were necropsied 90 to 240 days later. Guinea worms were recovered from 10 (56%) of 18 ferrets. A total of 44 worms were recovered, for an average of 4.4 worms per infected ferret. Gravid female worms were recovered as early as 128 days postinoculation. Thirteen (87%) of 15 gravid female worms were recovered from the extremities. Living male worms were recovered at 200 days of age, indicating that not all male worms die shortly after mating. First-stage larvae recovered from gravid females as early as 200 days of age were found to be infective to the copepod, Acanthocyclops vernalis. These observations suggest that the ferret is an excellent laboratory animal for dracunculiasis research.