An electron microscope study of the tegument of Hunterella nodulosa Mackiewicz and McCrae, 1962 (cestoidea: Caryophyllidea)

Ultrastructural observations using transmission and scanning electron microscopy reveal the tegument to be basically similar to that of other cestodes. The syncytial distal cytoplasm is devoid of organelles except for rod-shaped bodies, believed to be secretory vesicles, and lamellated bodies which probably contribute the raw material for new microtriches. There is evidence that these vesicles originate from the Golgi found in the sub-cuticular cells.Three types of microtriches are described: typical ones with well-developed spines, ones with short filaments instead of spines, and ones with no spines. Microtriches with spines are found only on the anterior part of the worm and may serve to anchor the worm. Microtriches on the posterior have no spines and are believed to be primarily involved in the absorption of nutrients. Between these two regions there is a transitional zone where all three types of microtriches can be found. In general the microtriches are quite uniformly distributed throughout the surface of the worm. The presence of cestodarian-like microtriches raises interesting evolutionary questions.Histochemical tests localized acid and alkaline phosphatase activity on various parts of the tegument, as well as on host intestine, while acian blue tests showed that acid mucopolysaccharide levels correspond with the concentration of the tegument vesicles.     

Hymenolepis microstoma: Ultrastructural localization of adenyl cyclase in the tegument

Peroxidase and phosphatase activities have been reported to be localized in the tegument of adult hymenolepidid tapeworms. In order to localize adenyl cyclase (EC 4.6.1.1) activity in the tegument of mature and gravid sections of Hymenolepis microstoma, 5-adenyl-imidodiphosphate was used as a substrate, and lead was used as a capturing agent. Results indicate that adenyl cyclase activity is present in the crypts between the microtriches of the mature sections and that activity is absent from the gravid sections.     

Tegument and apical end organ fine structure in the metacestode and adult Proteocephalus ambloplitis

The tegument of plerocercoid and adult P. ambloplitis was examined. Differences in tegument structure existed between these two stages. Plerocercoids of P. ambloplitis lacked extensive vacuolization and unicellular gland cells characteristic of adult tegument. Plerocercoid microtriches were short and conoid; adult microtriches were lenticular with an extended, whip-like shaft. An inclusion, not previously reported from proteocephalid cestodes, is described. Adult tegument had ducts, originating from underlying unicellular glands, extending through the distal cytoplasm and opening to the exterior between microtriches.

The apical end organ cavity of P. ambloplitis contained numerous labyrinth-like spherical bodies. These structures appeared to be synthesized and secreted into the end organ by a thin cellular lining of the end organ. This lining was composed of discrete, filamentous cells believed to be modified subtegumental cell bodies. Spherical structures identical to those observed within the end organ cavity occurred within this cellular lining. The spherical bodies may be associated with enzymes necessary for tissue migration by the metacestode.     

The forebody glands and surface features of the metacercariae and adults of Microphallus similis

Numerous unicellular glands are present in the forebody of the metacercariae and adults of Microphallus similis. The gland cell bodies lie beneath the tegumental perinuclear cytoplasm and ducts pass upwards penetrating the tegumental distal cytoplasm. Electron dense granules are secreted onto the forebody surface and into the oesophagus via small pits in the tegument. Histochemical tests revealed that the glands contain diastase-resistant neutral mucosubstances, RNA, protein, esterase and small amounts of acid phosphatase; inhibitor studies indicated that the esterase was acetylcholinesterase (EC 3.1.1.7). Ultracytochemistry showed that the mucosubstances were located in the general cytoplasm of the gland cells and that cholinesterase was present in the granules. The forebody of M. similis is covered by large toothed spines whereas the hindbody only bears short peg-like spines. It is suggested that the forebody spines may have an irritating, abrasive effect on the host's intestinal mucosa and that the secretion of acetylcholinesterase in the region of these spines may produce a ‘local anaesthetic’ effect on the host's gut by reducing movement in the immediate vicinity of the fluke, so decreasing the likelihood of expulsion.     

Taenia taeniaeformis (Cestoda) in the rat: ultrastructure of the host-parasite interface on days 8 to 22 postinfection

The host-parasite interface was examined at the ultrastructural level 8 to 22 days postinfection (DPI) with metacestodes of Taenia taeniaeformis in the rat. Throughout this phase of development the parasite surface was invested with a dense surface coat of complex microtriches. At 8 to 14 DPI the plasma membrane of each microthrix extended beyond the distal end of the electron-dense tip, forming a slender tubular streamer over 10 microns long; by 18 DPI these had shortened and withered. Host cell processes interdigitated with the microtriches without evidence of harm to the parasite surface or the underlying tegument. The cells, on the other hand, became damaged, and their contents were shed into the matrix surrounding the microtriches. Lipid inclusions appeared within the parasites, and in the cytoplasm of surrounding inflammatory cells. By 22 DPI fibroblastic activity had resulted occasionally in the formation of a capsule surrounding a free-floating cysticercus, while in others intense granulocytic infiltration persisted with abutment and intermeshing of host cell and parasite surface processes; however there was still no evidence of any adverse effect on the microtriches, though many granulocytes were clearly pyknotic and degenerating. Evidently, the vigorous cellular response of the host is ineffective in either containing the expansion of the parasite or compromising the integrity of its surface membrane. The changing characteristics of the microtriches may be related to the need for dissolution of both intercellular matrices, and host cells as the vesicular organism rapidly increases in volume.     

Fasciola hepatica: Histochemical observations on juveniles and adults and the cytopathological changes induced in infected mouse liver

The histochemical characteristics of juvenile intrahepatic forms of Fasciola hepatica have been compared with those of the adults. The histochemical distributions of carboxylesterase (EC 3.1.1.1), acetylcholinesterase (EC 3.1.1.7) and alkaline phosphatase (EC 3.1.3.1) were similar in both juvenile and adult forms. Differences were apparent in occurrence and distribution of β-glucuronidase (EC 3.2.1.31) and N-acetyl-β-glucosaminidase (EC 3.2.1.29) activities in the tegument, parenchyma, and caecal cells. These may reflect the dietary mode and relationship to the host of the juvenile intrahepatic and adult bile-duct forms. Starvation of both juveniles and adults is accompanied by an increase in the granule-associated staining reactions for acid hydrolases in some of the parenchymal cells with a concomitant decrease in staining for glycogen in these cells. This response to starvation is reversible with refeeding, indicating that it is probably a genuine response to nutrient stress and not to degeneration induced by the in vitro conditions of the flukes. Carboxylesterase and acid phosphatase (EC 3.1.3.2) have been demonstrated to be polymorphic, using polyacrylamide disc electrophoresis. Changes were observed in the staining patterns with starvation. The juvenile flukes stimulated a considerable increase in the level of lysosomal hydrolases and alkaline phosphatase in mouse hepatic cells.     

Schistosoma haematobium: histochemistry of glycogen, glycogen phosphorylase a and glycogen branching enzyme in niridazole-treated females.

The body posterior to the ovary of Schistosoma haematobium females was investigated. Glycogen, glycogen phosphorylase a (EC 2.4.1.1) and glycogen branching enzyme (EC 2.4.1.18) activities were detected in the subtegumental muscle system, parenchyma and mature vitelline cells, whereas no activities were detected in the tegument and immature vitelline cells of the parasite. Administration of a single niridazole dose of 250 mg kg-1 to the pouched mouse (Saccostomus camestris) produced the following changes in S. haematobium females: a relatively rapid depletion of glycogen stores due to disruption of the absorptive surface of the parasite, and to an increase in the activity of glycogen phosphorylase a; a reduction in the phosphorylase a to phosphorylase b-conversion capacity of glycogen phosphorylase phosphatase (EC 3.1.3.17); a decrease in glycogen branching enzyme activity; and a relatively rapid degeneration of parasite cells possibly due to their loss of endogenous energy reserves.     

The ultrastructural localization of acetylcholinesterase activity in the scolex of the cysticercus of Hydatigera taeniaeformis

Acetylcholinesterase activity in the scolex of the cysticercus of Hydatigera taeniaeformis was studied by means of electron microscopy. Acetylthiocholine iodide was used as the substrate and 10(-4) M of eserine used to inhibit acetylcholinesterase activity. The sites of activity were visualized within the microtriches of the distal cytoplasm, muscle bundles, cytoplasm, vesiculated bodies, flame cells and hooks of the rostellar region. Enzyme activity was also localized within the nuclear membrane, neuroplasm, and membranes of nerve cells.     

鱊头槽绦虫原尾蚴皮层的超微结构观察

利用透射电镜观察了头槽绦虫 (Bothriocephalusacheilognathi)原尾蚴皮层的超微结构。头槽绦虫原尾蚴的皮层为典型的合胞体结构。皮层表面有浓密的微毛与少量的突起。与成虫相比 ,原尾蚴的微毛长而且粗 ,呈现单态性 ,推测在进入终末宿主的过程中有脱落和再生现象。突起分布在头部与体侧 ,含有较少的电子致密颗粒。在原尾蚴皮层细胞质中观察到三种腺细胞的分泌过程 ,即顶分泌、外分泌和微分泌过程。原尾蚴身体后部观察到尾钩和穿刺腺管道的开口。在核周区和纤维层下可见发育较好的环肌与纵肌。本文还对原尾蚴皮层突起的功能进行了探讨。     

Histochemical studies on Raillietina (Raillietina) johri (Cestoda: Davaineidae). I. Nonspecific and specific phosphatases.

Certain phosphatases have been localized by histochemical techniques in various tissues of a pigeon cestode, Raillietina (Raillietina) johri. Acid phosphatase (AcPase), alkaline phosphatase (AlPase) and adenosine triphosphatase (ATPase) were present in almost all structures: tegument; subtegumental muscles; subtegumental cells; excretory canal; testes; sperm ductules; vas deferens; cirrus sac; cirrus; ovary; receptaculum seminis; vagina; vitelline gland cells; oocytes; uterus; embryonated eggs. AlPase was absent in parenchyma, spermatocytes, spermatids and spermatozoa. AlPase activity was more intense in the tegument of mature gravid proglottides. AcPase and ATPase were visualized in various stages of spermatogenesis of the parasite. ATPase activity was also observed in chromosomes. 5'-nucleotidase (AMPase) activity was restricted to embryonated eggs only. Functional significance of these phosphatases is discussed.     

Ultrastructure of the cirrus sac of echinophallid tapeworms (Cestoda, Bothriocephalidea) and the terminology of cirrus hard structures

Transmission (TEM) and scanning (SEM) electron microscope methods were used to study the fine structure of the cirrus, cirrus sac, internal seminal vesicle, ejaculatory duct, prostate glands and cirrus armature of Echinophallus wageneri (Monticelli, 1890) and Paraechinophallus japonicus (Yamaguti, 1934) (Bothriocephallidea: Echinophallidae). The cirrus sac of these species has two unique ultrastructural features: a thick wall with two bands of muscles and prominent, rooted hard structures. Rare traits echinophallids share with diphyllobothriideans are microtriches on the ejaculatory duct and with spathebothriideans, well-developed unicellular prostate glands outside the cirrus sac. Because there is a similarity of cirrus armature and rostellar hooks in having a tegumental localisation and in having a heterogenous structure of the blade and root, a cortex, a central pulp region and a recurved apex, these structures are named “modified hooks” instead of spines. They also have a spiral arrangement; no base plate was observed. True spines, as found in trematodes, are between the surface and basal plasma membrane of the external syncytial layer of the tegument, rest on the basal plasma membrane of the distal epithelial cytoplasm, show a homogeneous electron-dark crystalline appearance and are covered by the surface plasma membrane. Aside from the characteristic hooks on the scolex of various cestodes, we see no evidence that would preclude the development of still other specialised structures, such as these modified hooks, from microtriches. In spite of the absence of studies on the development of modified hooks from the cirrus of echinophallids and/or its consideration as derived from microtriches, we assume that like microtriches, formation of modified hooks is from tegumental bodies and therefore they are derivative structures of the cestode tegument.     

Histochemical findings of the tribocytic organ and tegument of Fibricola seoulensis

The tribocytic organ and tegument of Fibricola seoulensis were examined histochemically for the detection of carbohydrates, mucosubstances, amyloid, collagen and alkaline phosphatase. The surface, secretes, gland cells of the tribocytic organ, and the tegument of the worms were positive to periodic acid Schiff (PAS) and PAS with diastase stain but negative to other stains. It was inferred that the tribocytic organ and tegument of F. seoulensis comprise neutral mucopolysaccharides, which may take a protective role against host enzymes. The surface and secretes of the tribocytic organ, and the tegument of the worms were also positive to double bridge PAP for alkaline phosphatase. This fact suggests that they may play a role as both self protective and host tissue lytic functions.     

Microtriches of tetraphyllidean metacestodes from Western Mediterranean striped dolphins (Stenella coeruleoalba)

The tegumental structures of two types of tetraphyllidean plerocercoids and two types of merocercoids (Phyllobothrium delphini and Monorygma grimaldii) from Mediterranean striped dolphins, Stenella coeruleoalba, are described for the first time using scanning electron microscopy. The tegument of all of the specimens was fully covered with microtriches. Four basic types were found: filitriches, blade-like spinitriches of different sizes and shapes, cone-shaped spinitriches (with two parallel small projections of equal length at the apex), and crowned cylindrical spinitriches (with 6-7 small papillae forming a crown at the apex); the two latter types are newly described. The two types of plerocercoids had a similar morphology and distribution of microtriches; in addition, cilium-like projections appeared interspersed among the microtriches on the apical sucker, accessory suckers, and distal bothridial loculus. Merocercoids exhibited a greater variety of tegumental structures, especially on the distal bothridial loculus. Both merocercoid types had regularly spaced papillae or "buttons" on the accessory suckers and the distal bothridial loculus that were composed of a central cilium-like projection surrounded by numerous filitriches. However, crowned cylindrical spinitriches were specific to P. delphini and cone-shaped spinitriches were specific to M. grimaldii. Differences in the morphology and distribution of scolex microtriches of adult cestodes have been considered useful for species identification. A previous molecular study has suggested that P. delphini and M. grimaldii are actually different congeneric species. Our study has shown that significant differences in the morphology and distribution of microtriches occur between these species at the merocercoid stage.     

Ultrastructure of the tegument of the cestode Paraechinophallus japonicus (Bothriocephalidea: Echinophallidae), a parasite of the bathypelagic fish Psenopsis anomala

Abstract. The ultrastructure of the tegument in Paraechinophallus japonicus (Bothriocephalidea: Echinophallidae), a cestode parasite of the bathypelagic fish Psenopsis anomala , was studied using scanning and transmission electron microscopy. Paraechinophallus japonicus lacks a true scolex. Four different types of microtriches have been observed on the tegumental surface of P. japonicus. Capilliform (∼2.3-μm long) and blade-like spiniform (∼1.4-μm long) microtriches are intermingled on the surface of the pseudoscolex. Capilliform microtriches are distinct in possessing a short base and a long electron-lucent cap. The strobila is covered with two types of microtriches, namely filiform (∼2.1-μm long) and tusk-shaped microtriches (≤4.5-μm long). Tusk-shaped microtriches are limited to the posterior border of each proglottid and are characterized by a short and narrow base, and a large and wide, sharply pointed, electron-dense cap. Similar tusk-shaped microtriches were previously found in members of the family Echinophallidae and may represent an autapomorphy of echinophallid cestodes, all of them being parasitic in centrolophid fish. A unified terminology of microthrix parts is proposed.     

Proliferation and metastases formation of larval Echinococcus multilocularis. II. Ultrastructural investigations

The larval stage (metacestode) of Echinococcus multilocularis was studied by means of electron microscopy (SEM, TEM) before and after subcutaneous transplantation to jirds (Meriones unguiculatus) and in their lymph nodes and lungs with parasite metastases. It was found that the metacestode consists of a network of solid, cellular protrusions (buds) of the germinal layer which transform to tube-like and cystic structures devoid or with a laminated layer. Proliferation of the metacestode apparently occurs by protruding filamentous solid cell columns (buds) from the germinal layer. Their tips have diameters of only one cell: they are covered with a smooth syncytial tegument without microtriches and are filled with undifferentiated cells which contain a nucleus with a large nucleolus. The tegument is constantly enlarged by fusion with the underlying undifferentiated cells that divide repeatedly. At some distance from the tip a cavity develops inside the protrusion, thus finally giving rise to a tube-like structure which may transform to a cystic expansion. Simultaneously, the surface of the protrusion changes by the formation of microtriches and the occurrence of an amorphous laminated layer. The latter is concentrically covered by connective tissue cells and large amounts of collagen. Within cyst-brood capsules, finally protoscoleces are formed from accumulations of undifferentiated cells beneath the tegument. The study has unequivocally proven the cestode nature of the cellular protrusions, and it is assumed that detachment of cells from these structures and their subsequent distribution via the circulation may play a role in the formation of metastases. The origin of the laminated layer is discussed.     

Morphological studies on the early development of Taenia taeniaeformis larvae in susceptible mice

Bortoletti G. and Ferretti G. 1985. Morphological studies on the early development of Taenia taeniaeformis larvae in susceptible mice. International Journal for Parasitology15: 365–375. Taenia taeniaeformis larvae which develop into infective strobilocerci in C3H mice have been studied from the 5th to the 15th day of development (L5–L15), both at light and electron microscope level. The L5 were initially compact, without a central cavity but then become vacuolized. Until stages L7–L8 they were surrounded by a perilarval amorphous layer (PAL) made up of a finely granular material which prevented the host cells from making contact with the larval tegument. The larval volume increased considerably between stages L6 and L8, remained unchanged from L9 to L13, but continued to increase thereafter. The larval cellular layer, which appeared as a single, large ‘syncitial system’, grow until stages L14–L15 when the scolex anlagen began to form. The tegument was initially incompletely organized and was covered by microvilli. These were completely replaced by microtriches from stage L8 onward. Sometimes both microvilli and microtriches were together observed in stage L7. Microvilli fragments, sometimes beaded, could be observed at L5 within the damaged cytoplasm of host cell debris. Very often they were branched at different heights, especially in stages L5–L7. In L10–L15 all undamaged microtriches increased in density and formed bundles which invaded the host cells. In stages L5–L8 and in some L9, muscular bundles started to become organized inside the tegumental distal cytoplasm (TDC), and after become independent in the subtegumental cellular layer (SCL). Until L8–L9 the larvae were surrounded by host cells debris. From stages L8–L10 onwards the adjacent host cells were less damaged though the larval microtriches penetrated them deeply. In stages L5–L7 neutrophils together with macrophages and some damaged hepatocytes were detected, while eosinophils were present only from L8 onward. In the other stages neutrophils clearly diminished in numbers, whereas macrophages had increased. No mastcells and few plasma cells were observed.     

扩张莫尼茨绦虫(圆叶目:裸头科)节间腺的超微结构及组织化学

用光学显微镜和透射电子显微镜观察了扩张莫尼茨绦虫节间腺形成过程的精细结构及一些组化变化。结果表明：节间腺是扩张莫尼茨绦虫皮层的特化部分,由节片后缘的皮层及其邻近细胞体向绦虫实质组织中陷入开始其形成过程,随着虫体发育的进行,新的陷入不断形成,原陷入的部分不断脱离皮层形成簇状腺体结构。节间腺的数目随着体节的发育不断增加,幼节中仅有少数几个(6～9个),而远端的孕节中多于100个。电镜下可见腺细胞体由细胞质管与腺皮层相联,簇状腺体结构为一合胞体形态,腺细胞体围绕并开口于椭球体或不规则形状的皮层腔中。离腺皮层远的腺细胞体电子密度高并含有与腺皮层相应的典型分泌颗粒,而靠近腺皮层的腺细胞体电子密度低,所含分泌颗粒较少。扩张莫尼茨绦虫节间腺的组化性质尚不完全清楚。糖与蛋白质等组化结果不稳定,随染液pH值及染色时间的变化等多种因素而改变。基于我们的研究及其他研究者的观察表明,节间腺可能参与外源基质形成虫卵的转运,同时他们可能在虫体节片脱落及虫卵溢出时起作用。     

Schistosoma mansoni: Radioautography of colchicine's effect on [3H]proline incorporation into adults in vitro

Adult Schistosoma mansoni were studied radioautographically in order to ascertain the effect of exposures to a fixed concentration of colchicine (5 × 10^?4M) for varying time intervals upon the incorporation of [³H]proline in the tegument. Additionally, a study was made on the effect of varying time exposures of colchicine on the cytochemical localization of alkaline phosphatase (EC 3.1.3.1) in the tegumental invaginations. Worms exposed to colchicine for more than 2 hr preceding addition of the labeled amino acid displayed significant changes in the pattern of distribution. The most profound change was noted in the male tegument where a statistically significant decrease was observed in treated worms. Female worms, on the other hand, failed to display any effect of the drug on the distribution pattern for the times utilized. The distribution of alkaline phosphatase activity was much reduced in the teguments of both sexes. Morphological effects of the drug included disappearance of microtubules from the cytoplasmic connectives, a stacking of RER in the subtegumental cells, and accumulation of discoid granules and membranous bodies in the subtegumental cells. It is hypothesized that the amino acid is associated with the discoid granule at the subtegumental cell level and is ultimately translocated, with the aid of microtubules in the cytoplasmic connectives, to the tegument. Alkaline phosphatase activity is assumed to be associated with the membranous body.     

Turnaround of Axoplasmic Transport of Selected Particle-Specific Enzymes at an Injury in Control and Diisopropylphosphorofluoridate-Treated Rats

: Reversal of direction (turnaround) of axonal transport of particle specific enzyme activities was studied at a ligature placed on rat sciatic nerve. In the principal experiment, the ligature remained on the nerve in vivo several hours, allowing enzyme activities (acetylcholinesterase, acid phosphatase, and monoamine oxidase) to accumulate immediately proximal to the tie. The nerve was then tied a second time, proximal to the first tie, and incubated in vitro for several more hours. Accumulation of enzyme activities just distal to the second tie was measured. This second accumulation, of activities traveling in the retrograde direction, was shown to be the result of turnaround in several ways. (1) The increase in activity distal to the second tie was equal to the decrease in activity proximal to the first. (2) The increase in enzyme activities distal to the second tie was greatly reduced when the accumulation proximal to the first tie was trapped by placing a third tie between the first and second ties. (3) It was shown that the activity that accumulated distal to the second tie could not have been in retrograde motion at the time of the first tie. (4) Accumulation distal to the second tie was not a function of the length of nerve segment included between the two ties. In contrast to the consistent occurrence of turnaround of orthograde flow, turnaround of retrograde flow could not be demonstrated. Turnaround transport was blocked by incubation in the cold and in the presence of NaCN or vinblastine. The turnaround process operated on all three enzymes studied, suggesting that it operates on lysosomes and mitochondria, as well as on the endoplasmic reticulum-like material bearing acetylcholinesterase. Evidence for the participation of the transport process in the renewal of AChE in the distal portions of the axon was obtained in experiments using diisopropylphosphorofluoridate and cycloheximide.