Surface ultrastructure of the advanced third-stage larvae of Gnathostoma nipponicum

The surface ultrastructure of advanced third-stage larvae (AL3) of Gnathostoma nipponicum was studied using scanning electron microscopy. The larvae were recovered from the grass snake Rhabdophis tigrina in the Republic of Korea. Parasites had a globular head bulb with a pair of lips at the anterior end and 2 labial papillae and an amphid on each lip. The head bulb was characteristically armed with 3 transverse rows of hooklets, averaging 36, 38, and 43 in number, increasing posteriorly. A total of 213-232 minute unidentate cuticular spines were present along the entire length of the larvae, forming the transverse striations. Two pairs of cervical papillae were located between the 8th and 12th transverse striations, and a pair of body papillae was seen laterally on the posterior third of the body. A pair of caudal phasmids was recognized near the posterior extremity. The surface ultrastructure of AL3 of G. nipponicum is unique compared with that of other species.     

The in vitro effect of albendazole, ivermectin, diethylcarbamazine, and their combinations against infective third-stage larvae of nocturnally subperiodic Brugia malayi (Narathiwat strain): scanning electron microscopy.

Scanning electron microscopy (SEM) was employed to observe the effects of ivermectin (IVM), diethylcarbamazine (DEC), and albendazole (ALB) alone, and the drugs in combination (ALB+IVM and ALB+DEC) against infective third stage larvae (L3) of nocturnally subperiodic (NSP) Brugia malayi (Narathiwat strain) in vitro. IVM, at a concentration of 10(-4) M, killed L3 within 1-2 h. The SEM data showed damage to the L3 surface and loss of regular cuticular annulations. The cuticles were grooved in the middle region of the body. In comparison with normal L3 before treatment with IVM, the cuticular surface showed transversed striations with periodic annulations. The result demonstrated that IVM showed a larvicidal activity against L3 of NSP B. malayi cultivated in vitro. Compared with those larvae in the control group, the treated larvae had no morphological changes in the cuticular surface at the head, body, and tail regions after cultivation with all drugs alone, and in their combinations at a concentration of 10(-5) M for 7 d. In this system, and at that concentration, only the larvae cultured with ALB alone remained highly motile. Although no morphological changes had been observed by SEM, those drugs used alone (IVM and DEC) and in combinations (ALB+IVM and ALB+DEC), reduced larval motility throughout the experiments at a concentration of 10(-5) M. The minimum lethal concentration (MIC) of IVM against NSP B. malayi was 10(-4) M.     

Larval Gnathostoma hispidum detected in the red banded odd-tooth snake, Dinodon rufozonatum rufozonatum, from China

A total of 205 larval gnathostomes were collected from 18 (22.5%) of 80 red banded odd-tooth snakes, Dinodon rufozonatum rufozonatum, which had been smuggled from China and confiscated at Customs in Busan, Republic of Korea. In order to identify the species, some of the larvae were observed by a light microscope and a scanning electron microscope (SEM). The larvae were 2.18 x 0.29 mm in average size, and had a pair of lips at the anterior end, a muscular esophagus, 2 pairs of cervical sacs, and brownish intestines. The head bulb was characteristically equipped with 4 rows of hooklets; the average number of hooklets in each respective row was 38.6, 40.5, 41.5, and 43.7. In SEM views, the mouth evidenced a pair of lateral lips of equal size in a half-moon shape. Each lip featured a couple of labial papillae and a small amphid located between the 2 papillae. The hooklets on the head bulb had single-pointed, posteriorly-curved tips. The cuticular spines were larger and more densely distributed on the anterior part of the body, and decreased gradually in size and number toward the posterior body. On the basis of these morphological characteristics, the larvae were identified as the third stage larvae of Gnathostoma hispidum.     

Moravecia australiensis n. g., n. sp. (Dracunculoidea: Guyanemidae) from the gills of the green porcupine fish Tragulichthys jaculiferus (Cuvier) in Australia

A new dracunculoid genus and species, Moravecia australiensis, is described from gill-filaments of the green porcupine fish Tragulichthys jaculiferus (Cuvier) (Tetraodontiformes: Diodontidae) from Moreton Bay, Queensland, Australia. Abundant mobile larvae and a few adult males with females occurred in the gill-filament between the epithelial basement membrane and efferent artery. Gills of all 69 fish examined contained larvae. Eleven harboured adult nematodes of a previously undescribed species belonging to the family Guyanemidae. The new species is placed within a newly proposed genus because it differs from the four existing genera in the family in possessing fine cuticular transverse striations, two forward protruding cephalic elevations, a circumoral elevation, a small triangular mouth surrounded by six cephalic papillae arranged in two lateral clusters of three each and a pair of large lateral amphids. Males have two pairs of pedunculate caudal papillae supporting the caudal alae. A key to the genera of the Guyanemidae is presented.     

Identification of human hookworm in failed-treatment cases using Chinese hamsters (Cricetulus griseus) and scanning electron microscopy

An attempt was made to identify the human hookworm involved in failed-treatment cases using abnormal hosts and scanning electron microscopy. Thirty-seven, 2 to 6 month old Chinese hamsters (Cricetulus griseus) from a closed, outbred, conventional colony, were each given between 20 and 120 filariform larvae per os. The larvae were cultured from faeces from mebendazole (Vermox) 500 mg single-dose, failed-treatment cases living in the lowveld farming area of the Transvaal Province, South Africa. About 60 to 78 days after inoculation, the animals were killed and adult worms were removed from their small intestines. Eleven (30%) of the 37 hamsters harboured a total of 31 adult worms (19 males and 12 females), while 26 hamsters were refractory to infection. The greatest number of worms recovered from a single animal was six. A total of 27 worms (17 males and 10 females) were subjected to examination by scanning electron microscopy. Micrographs showed male and female worms to be morphologically all of the Necator americanus species, as identified by a pair of ventral and dorsal cutting plates, a dorsal tooth and the fused terminus of spicules in the male bursa. The transverse cuticular striations were distinct and smooth. Several points of interest arose from the results of this study and are discussed.     

Nematode cysts and larvae found in Achatina fulica Bowdich, 1822

This study describes the morphology of the nematode cysts and larvae found in Achatina fulica (giant African snail) in Brazil. Sixty snails were collected in Mesquita, Rio de Janeiro State. Fourteen of the snails were naturally infected. The cysts were spherical, pink colored and measured 0.97 to 1.57 mm in diameter. In the majority of cases they had a single larvae involved in amorphous material. A total of 222 encysted larvae were recovered. Of these, 30 were utilized in the morphological study. The length of the larvae varied from 2.57 to 5.8 mm and they were classified as small - up to 3.5 mm; medium - from 3.53 to 4.5 mm; and large - greater than 4.52 mm. The average length of the larvae in the three groups was 2.85, 3.87 and 5.23 mm, respectively. The larval cuticle was white, shiny and transversally striated until the posterior end of the body. At the anterior end there is a mouth with three lips, with amphids and papillae, followed by a muscular esophagus with average length of 0.61 mm, terminating in an esophageal bulb and having a nerve ring in the middle third of the esophagus, and an intestine with an opening near the posterior end. The tail begins from this opening and has two types of ends: short and abrupt or long and gradually tapering. The difference in the tail end can suggest sexual dimorphism, although no primordial reproductive structures were observed. These characteristics were not sufficient to identify the larvae, so there is a need for further study.     

Cuticular morphology of five species of Dirofilaria: a scanning electron microscope study

Cuticular markings of five species of Dirofilaria were examined by SEM. Variations in the patterns on the en face view were observed among the species. Transverse striations were observed on the cuticles of all five species, but longitudinal ridges were lacking on Dirofilaria immitis, the type species of the subgenus Dirofilaria. They were present on the other four species, Dirofilaria repens, Dirofilaria tenuis, Dirofilaria corynodes, and Dirofilaria magnilarvatum, members of the subgenus Nochtiella. Longitudinal ridges and other cuticular structures and patterns were seen to vary among the species and with the portion and aspect of the worm examined. It is suggested that SEM would be a good tool for a detailed study of specific cuticular morphology of all species of Dirofilaria, thus contributing toward their differentiation and recognition.     

A new species of camallanidae from Ageneiosus ucayalensis (Pisces: Siluriformes) from Pará State, Brazil

Procamallanus (Spirocamallanus) belenensis n. sp. is described using light and scanning electron microcopy, based on specimens taken from Ageneiosus ucayalensis, Castelnau, 1855 (Pisces: Siluriforme), a freshwater fish commonly called mandubé, in Pard State, Brazil; Procamallanus (Spirocamallanus) rarus is recorded from the same host. To our knowledge there are no previous reports of nematodes from this fish species. Procamallanus (S.) belenensis is characterized by the presence of 8-13 cuticular spiral thickenings coating 2/3 of posterior region of the buccal capsule of males and females (the anterior third is smooth); small sub-equal spicules; 3 and 4-5 pairs of pre- and post-cloacal papillae, respectively; the proportion of muscular and glandular esophagus is 1:1.5-2; males present 2 spike-like projections at the tail tip, which are absent in females by light microscopy. The most closely related species are Procamallanus (Spirocamallanus) inopinatus and Procamallanus (Spirocamallanus) pintoi. The new species differs from the most closely related species by the presence of 8-10 cuticular spiral thickenings in the buccal capsule of males and 9-13 in the females, 8 pairs of male clocal papillae (3 pre- and 5 post-cloacal, respectively), and only males bearing 2 minute, spike-like projections at the posterior end.     

屋尘螨成螨形态的扫描电镜观察

【目的】利用扫描电子显微镜（scanning electron microscopy, SEM）观察屋尘螨Dermatophagoides pteronyssinus颚体、 躯体、 外生殖器及足等的形态结构。【方法】从床尘、 枕尘中采集屋尘螨, 分离出雌雄成螨, 在体视显微镜下清洗处理活螨后, 用SEM观察其外部形态特征。【结果】SEM照片显示, 屋尘螨螯肢钳状, 须肢扁平; 体表具细密皮纹, 似指纹状, 纹间距小于2 μm。外生殖器位于腹面正中, 雌螨为产卵孔, 雄螨生殖孔具1对叶状生殖盖。肛门呈纵行裂孔, 雄螨具2个肛吸盘。雌螨足跗节末端各具爪垫1个, 雄螨跗节Ⅳ具2个吸盘。【结论】本研究观察结果为尘螨鉴定提供了更多依据。     

Observations on the Morphology of the Pine Wood Nematode, Bursaphelenchus xylophilus

Scans of slash pine wood chips infested with pine wood nematodes showed coiled aggregates of 5-10 nematodes in the axial resin canals, and 1-2 nematodes folded in the radial resin canals. Observations of the pine wood nematode by SEM showed the head to be offset with six labial lobes. A smaller circle of six inner lobes surrounded the stoma. A single labial sensillum was observed on each of the two subventral and two subdorsal lobes. The two opposite lobes, without sensilla, bore a pore-like amphid. The female vulva flap was observed to be a pouch. In addition to two pairs of copulatory papillae previously described for the male, three additional papillae were observed: an additional post-anal pair, and a single papilla anterior to the cloaca. Spicules were fused ventrally. Distal spicule ends terminated in two fused half discs. Caudal alae were angular at the tail terminus, but culed inward at the corners to appear oval. Lateral fields had four lateral striations.     

文昌鱼Hedgehog基因敲除和突变体表型分析

文昌鱼隶属脊索动物门头索动物亚门,是无脊椎动物到脊椎动物的过渡类群,其躯体结构简单,是研究胚胎发育的理想材料。本文以文昌鱼为实验对象,利用TALEN敲除技术对Hedgehog(Hh)基因在胚胎发育中的功能进行了研究。在文昌鱼Hh基因翻译起始位点下游附近选取TALEN目标位点,根据此序列组装相应TALEN重组质粒,体外合成mRNA,向未受精卵注射mRNA后,经体外受精获得F₀代胚胎。效率分析显示,靶向该基因的TALEN mRNA可导致F₀代胚胎在相应基因组区域发生突变的比例为34%。对部分F₀个体所产配子筛查发现,TALEN引起的突变可进入配子,将其中1尾突变类型为8 bp缺失的雄性个体与野生型雌性配对获得F₁群体,对F₁群体逐尾筛查,从中获得多尾携带8 bp缺失的杂合子;这些杂合子相互配对所产的F₂代胚胎,其中约有1/4个体在幼体早期出现躯体前端和尾向下弯曲、脊索前端腹侧的中胚层组织发育不全,不能开口等;随着幼体生长发育,躯体前端和尾部进一步卷曲,口部仍未形成,左右各形成一个口前窝,内柱和鳃裂位于躯体腹侧,最终因无口摄食而死亡。基因型分析发现,上述畸形胚胎均为Hh纯合突变体,其与杂合子及野生型比例分布符合孟德尔遗传定律,表明这些发育畸型的特征与Hh基因功能缺失有关。     

The ultrastructural characterization of the tegument of Clonorchis sinensis (Cobbold, 1875) cercaria.

The tegumental ultrastructure of the cercaria of the liver fluke, Clonorchis sinensis (Cobbold, 1875), was studied by scanning and transmission electron microscopy. The body surface is almost encircled by many rows of regularly arranged spines. The tegumental syncytium of the body contains many rod-shaped dense granules and central electron-lucent bodies, neither of which are present in the tail tegument. There are four rows of hooked teeth and modified spines on the oral cone. These teeth are differentiated morphologically and probably functionally from the other body spines. Disc-shaped papillae with long or short cilia are distributed on the body in a bilaterally symmetrical pattern dorsally and ventrally. There are 30 to 37 papillae with much longer cilia laterally. Four pairs of papillae were found on the tail. From their structure and location these papillae appear to have a mainly tango- or rheoreceptive function. Another type of sheathed papillae is situated around the oral sucker. The cuticular tegument is expanded laterally at the base of the tail, forming a characteristic sac-like structure.     

A gastric-brooding asteroid, Smilasterias multipara

The gastric-brooding asterinid sea star, Smilasterias multipara, broods from late August to early November in the shallow sublittoral zone of southeastern Australia. We observed males and females spawning in the laboratory. They shed gametes through gonopores on the sides of the arms. The eggs were orange, about 1.0 mm in diameter, and heavier than seawater. They were externally fertilized by sperm, and placed into the stomach of the female by the tube feet. Twenty-four hours after fertilization, the first cleavage occurred. Cleavage was equal, total, and radial. Development via a non-feeding lecithotrophic brachiolaria was direct, there being no planktrotrophic bipinnaria or brachiolaria larva. Embryos developed, through wrinkled blastula and gastrula stages, into brachiolariae with arms. All of the surfaces of the brachiolaria were covered by cilia. At metamorphosis, a starfish rudiment appeared on the posterior portion of the larval body, while the anterior portion of the larval body was absorbed. Two months after fertilization, metamorphosis was complete. After metamorphosis, juveniles in the stomach grew six pairs of tube feet in each arm. Juveniles, 3 mm in diameter, emerged from the mouth of the mother in early November. Developmental evidence suggests that this asteroid has evolved mechanisms for the protection of larvae and juveniles from gastric digestion.     

红棕象甲室内生物学特性及形态观察

在室内对红棕象甲Rhynchophorus ferrugineus Fab.生物学特性和形态进行系统观察。结果表明在26℃、RH80%、寄主食料华盛顿棕榈饲养条件下,红棕象甲世代历期平均114.77d,最短87.00d、最长166.00d,其中卵期平均3.50d,幼虫期平均36.68d,结茧到产卵前历期平均29.96d。幼虫以8龄为主,1~2龄幼虫的体重小于0.01g,头壳宽短于1.0mm,3龄后幼虫体重、头壳宽度、体长和体宽开始增加,8龄时幼虫平均体重为3.36g,头壳宽度6.0mm。成虫雌雄性比1.78∶1,补充营养能显著延长成虫寿命,在无补充营养时,雌雄成虫寿命只有7.00d左右,而在补充营养后,雌雄成虫寿命最长可分别达130.00d和244.00d。雌虫产卵量最大376粒,最小89粒,平均216粒。     

The cuticular proteins from free-living and parasitic stages of Haemonchus contortus--I. Isolation and partial characterization

1. Cuticles were isolated from the adult males, adult females, the second molt (2M) sheath from the infective larvae (L3(2M)), and the parasitic third stage (L3) of the sheep parasite Haemonchus contortus by a combination of mechanical disruption and detergent treatment. 2. The 2ME soluble cuticular proteins from adult males contained 4 or 5 major protein bands with molecular weights ranging from 100 to 56 kD with the most prominent band at 56 kD. The cuticular proteins from adult females were similar to the male. 3. Cuticular proteins from the larval stages, 2M cuticle, and L3 cuticle, differed from the adults and from each other. The most prominent protein bands were observed with molecular weights on 78 and 39 kD for the L3 cuticle and 100, 91 and 46 kD for the 2M cuticle. The 2ME soluble cuticular proteins from all developmental stages were at least partially digested by bacterial collagenase. 4. The amino acid composition of cuticular proteins was similar for the L3 and 2M, but adults had lesser amounts of glycine and greater amounts of basic amino acids than the larval stages. The amount of the isolated cuticle solubilized by the 2ME treatment was greatest in adults (80%) compared to the L3 (64%) and the 2M (22%). 5. These results support a hypothesis that there are quantitative and qualitative stage specific differences in the cuticular proteins of H. contortus.     

Biparental mouthbrooding of the catfish Phyllonemus filinemus in Lake Tanganyika

This article reports biparental mouthbrooding of the bagrid catfish Phyllonemus filinemus in Lake Tanganyika, based on analysis of specimens collected during SCUBA diving. This catfish was nocturnally active, and in the daytime it was concealed singly or in pairs beneath rocks. Within a breeding pair, the male or female alone incubated all the brood in the mouth until the offspring attained 12 mm or so in total length, but thereafter joint mouthbrooding or guarding by both parents took place. Most females of nonbrooding pairs showed high values of gonadosomatic index (GSI), whereas all females of brooding pairs and most single females showed low GSI values. This fact indicated that a pair is formed at a time near the gonadal maturation of the female and separates after the brood is reared. No significant difference in body condition among parents of different reproductive states was observed, which suggested that their condition does not deteriorate markedly as the result of foraging by an off-duty parent. Received: September 16, 2000 / Revised: November 18, 2000 / Accepted: January 23, 2001     

Larval development of Argentine hake Merluccius hubbsi

The ontogeny of the developmental stages of the hake Merluccius hubbsi is described. Fish larvae and post-transitional juveniles were collected in the Nor-Patagonian area from 1989 to 2004. The opening of the mouth and the pigmentation of the eyes are coincident with yolk resorption, finishing the yolk-sac stage. This species presents pigmentation on the head, trunk and tail typical of gadiform larvae. Pectoral fin development is completed during the transformation stage. The post-transitional juvenile stage begins when the fin-ray complements are complete and squamation begins. The fins become fully formed in the following sequence: pelvic fins, first dorsal fin, second dorsal and anal fins together, caudal fin and pectoral fins. The caudal complex is totally developed in larvae of 22·0–23·0 mm standard lengths ( L _S) and all vertebral elements are first observed in larvae of 8·5 mm L _S. The rate of development of M. hubbsi observed in this study could be faster than the rates reported for other species of Merluccius by different authors.     

广西发现版纳鱼螈白化幼体一例

2007年5月,在广西北流市六麻镇捕到一例雄性版纳鱼螈(Ichthyophis bannanicus)白化幼体。该个体完全白化。体重9.1 g,全长195 mm,尾长3.85 mm,体宽10.36 mm,吻长3.09 mm,尾鳍高5.71 mm,吻端至第一环沟9.47 mm,吻端至第二环沟13.41 mm,鼻间距2.42 mm,眼间距5.17 mm。与正常个体对比发现:该白化幼体的尾长、体宽、尾鳍高大于正常范围,吻端至第二环沟小于正常范围,其余数据均在正常范围内。     

土耳其斯坦东毕吸虫的扫描电镜观察

血吸虫类中如血居科(sanguinicolidae)的Aporocotyle simplex Odhner,1900、裂体科(Schistosomatidae)的日本血吸虫(Schistosoma japonicum)曼氏血吸虫(S.mansoni)及埃及血吸虫(S.haematobium)等多种血吸虫均经扫描电镜观察(Johnson and Moriearts,1969;Silk et al., 1969;Robson and Erasmus, 1970; Miller et al., 1972; Kuntz et al.,1976、1977;Voge et al., 1978; Thulin, 1980;及何毅勋和马金鑫,1980等)。关于土耳其斯坦东毕吸虫[Orientobitharzia lurkestanica(skrjabin,1913)Dut et Srivastava,1955]的体表扫描电镜尚无报告,而只见有此虫种体壁及肠管的透射电镜观察的资料(Lavrov and Fedoseenke,1978)。本文部分作者最近在内蒙东部兴安岭以南部分地区进行牛羊土