A histochemical study of the secretions of reproductive glands and of the egg envelopes of Achatina fulica (Pulmonata: Stylommatophora)

Histochemical investigations of the secretions of reproductive glands—albumen gland, apical uterus, basal uterus and prostate gland—indicate the presence of galactogen in the albumen gland, acid mucopolysaccharide in the apical uterus, and lipoprotein in the basal uterus and prostate gland of A. fulica. The proteinaceous secretions produced by the glands do differ in their terminal reactive sites. Intense alkaline phosphatase reaction is found in albumen gland and apical uterus; carbonic anhydrase activity could be detected mainly in the uterine glands. The cyclical secretory activity of the reproductive glands has been studied preparatory to egg-laying and in the spent phase. Histochemical characteristics of the egg envelopes—albumen, shell membrane and egg-shell—suggest a possible sequential deposition of glandular products during the descent of eggs through the repv. ductive tract. The factors contributing to the stability and resistant nature of the egg envelopes, and the possible role of nutritive materials contained in reproductive gland secretions in the development of the embryo (even while the eggs are inside the uterus), are discussed.     

Localization and characterization of acharan sulfate in the body of the giant African snail Achatina fulica

Acharan sulfate is a glycosaminoglycan (GAG), having the structure →4)-2-acetamido-2-deoxy-α- -glucopyranose(1→4)-2-sulfo-α- -idopyranosyluronic acid (1→, isolated from the body of the giant African snail Achatina fulica. This GAG represents 3–5% of the dry weight of this snail's soft body tissues. Frozen sections and polyester wax sections of the snail's body were stained by Alcian blue-periodic acid-Schiff's reagent (PAS) to localize acharan sulfate. Alcian blue staining indicated that GAG was mainly secreted into the outer surface of the body from internal granules. A highly mucous material was collected and treated and the acharan sulfate was recovered by ethanol and cetyl pyridinium chloride precipitation. Crude acharan sulfate was purified by DEAE-Sephacel ion-exchange chromatography. Depolymerization of intact mucus and purified acharan sulfate fractions by heparin lyase II (heparitinase I) from Flavobacterium heparinum produced an unsaturated disaccharide as a major product, establishing the repeating unit of acharan sulfate. These results demonstrate that mucus in the granule and secreted to the outside of the body is composed entirely of acharan sulfate.     

褐云玛瑙螺（Achatina fulica）胚胎发育不同时间酯酶同工酶分析

采用聚丙烯酰胺凝胶电泳法,分析了褐云玛瑙螺胚胎发育不同时间的酯酶（ＥＳＴ）同工酶。实验结果表明,在２５℃控温条件下胚胎发育早期ＥＳＴ同工酶谱带显示较弱;到初孵幼螺前的整个发育阶段的ＥＳＴ同工酶谱带呈逐渐递增趋势;初孵幼螺ＥＳＴ同工酶谱带增多。两种不同温度（在３０℃控温条件下进行了胚胎发育三个不同时间的ＥＳＴ同工酶分析）下胚胎发育至８小时的ＥＳＴ 同工酶谱带差异显著。初孵幼螺前２４小时的胚胎发育ＥＳ     

Studies on the Metabolism of Gallic Acid by Microorganisms

The intermediary metabolism of gallic acid by Aspergillus niger under the influence of some added inhibitors has been studied. The decomposition of gallic acid by lyophilized cells under fluoroacetate inhibition allowed cis-aconitic acid, α-ketoglutaric acid and citric acid to accumulate. A mechanism of gallic acid decomposition via cis-aconitic acid has been inferred.     

The secondary invasion of giant African land snail has little impact on litter or seedling dynamics in rainforest

In the absence of empirical evidence, invasive species are often assumed to have negative impacts because of their conspicuously high abundance. The giant African land snail Achatina (Lissachatina) fulica is one such invader where its impact in natural ecosystems remains completely untested. On Christmas Island (Indian Ocean), A. fulica has become established across large tracts of rainforest following the impacts of invasive yellow crazy ant (Anoplolepis gracilipes) in mutualism with non‐native scale insects. Yellow crazy ants facilitate the secondary invasion of A. fulica by extirpating native red land crabs (Gecarcoidea natalis) that are normally effective predators of A. fulica. We used a multifaceted approach to investigate some potential impacts of abundant A. fulica in invaded rainforest. Over the course of a wet season, diel activity transects showed that A. fulica consumed detrital material almost exclusively. However, stable isotope analysis did not confidently identify A. fulica as a predominantly detritivorous species. We found no statistically significant treatment effects of A. fulica exclusion on standing leaf litter and seedling recruitment processes during a 6‐month manipulative field study. However, litter cover and biomass did remain slightly higher where A. fulica were excluded, albeit with overlapping confidence intervals with control plots. Our study constitutes the first empirical test for impact of A. fulica in a natural ecosystem and suggests that for Christmas Island rainforest, this species is not a damaging invader. Other studies will need to assess the impacts of A. fulica in other natural areas before these findings could be considered broadly applicable.     

First record of a nematode Metastrongyloidea (Aelurostrongylus abstrusus larvae) in Achatina (Lissachatina) fulica (Mollusca, Achatinidae) in Brazil

Achatina (Lissachatina) fulica was introduced in Brazil in the 1980s for commercial purposes (“escargot” farming) and nowadays, mainly by human activity, it is widespread in at least 23 out of 26 Brazilian states and Brasília, including the Amazonian region and natural reserves, where besides a general nuisance for people it is a pest and also a public health concern, since it is one of the natural intermediate host of Angiostrongylus cantonensis, ethiological agent of the meningoencephalitis in Asia. As Brazil is experiencing the explosive phase of the invasion, the Laboratório de Malacologia do Instituto Oswaldo Cruz/Fiocruz has been receiving samples of these molluscs for identification and search for Angiostrongylus cantonensis and Angiostrongylus costaricensis larvae. While examining samples of A. fulica different nematode larvae were obtained, including Aelurostrongylus, whose different species are parasites of felids, dogs, primates, and badger. Morphological and morphometric analyses presented herein indicated the species Aelurostrongylus abstrusus, as well as the occurrence of other nematode larvae (Strongyluris-like) found in the interior of the pallial cavity of A. fulica. This is the first report in Brazil of the development of A. abstrusus infective larvae in A. fulica evidencing the veterinary importance of this mollusc in the transmission of A. abstrusus to domestic cats. Since the spread of A. fulica is pointed out in the literature as one of the main causative spread of the meningoencephalitis caused by A. cantonensis the authors emphasize the need of sanitary vigilance of snails and rats from vulnerable areas for A. cantonensis introduction as the port side areas.     

A single step purification of a sialic acid binding lectin (AchatininH) from Achatina fulica snail

A sialic acid binding lectin, Achatinin_H, was purified in single step from the hemolymph of the land snail, Achatina fulica, by the affinity chromatography on sheep submaxillary mucin coupled to Sepharose 4B. The yield of the lectin was found to be 3 mg from 100 ml of hemolymph. The homogeneity of the lectin was established by alkaline gel electrophoresis, immunodiffusion, immunoelectrophoresis and analytical isoelectrophoresis. The molecular weight of the native protein was 242000, having identical subunits of Mr 15000. The lectin agglutinated rabbit erythrocytes in the presence of Ca^2–. The inhibition study clearly suggests that the binding site of the lectin recognizes sialic acid as the immunodominant sugar. This was further confirmed by the observation that there was a marked decrease of agglutinating activity of the lectin with neuraminidase treated rabbit erythrocytes and asialofetuin was unable to inhibit the activity of Achatinin_H. Among the inhibitors used the glycoconjugate containing 2-6 linkages of N-acetylneuraminic acid with subterminal galactopyranose or 2-acetamido-2-deoxy-galactopyranose residue was found to be better inhibitor than that containing 23 linkages of N-acetyl neuraminic acid. Besides that sialoglycoprotein containing both N and O type of glycosidic linkages plays an important role in binding with the lectin. Fetuin was found to be the best inhibitor.     

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.     

四种入侵植物提取液对褐云玛瑙蜗牛的防控效果

实验室内观察了4种入侵植物水提取液对褐云玛瑙蜗牛成体和幼体的触杀及抑制蜗牛卵发育效果的差异性。结果表明:(1)以1.0 g/mL薇甘菊和南美蟛蜞菊水提取液对蜗牛喷杀处理, 对蜗牛触杀的有效时间分别为82 h和118 h;马樱丹触杀蜗牛的致死时间和有效时间最慢,4种入侵植物水提取液对成体褐云玛瑙蜗牛触杀效果强弱顺序为:薇甘菊 > 南美蟛蜞菊 > 五爪金龙 > 马缨丹。(2)1.0 g/mL 的4种入侵植物水取液对幼体蜗牛触杀的有效时间比成体蜗牛缩短2倍以上,幼体蜗牛个体死亡数量呈急剧增加趋势, 而成体蜗牛个体死亡数量呈相对平缓的增加趋势。(3)1.0、1.5 g/mL及2.0 g/mL薇甘菊水提取液对成体蜗牛触杀效果差异显著(P < 0.05)。以2.0 g/mL薇甘菊水提取液触杀蜗牛的有效时间最短, 与10%的四聚乙醛处理组相当, 为78 h, 显著少于1.5 g/mL的96 h和1.0 g /mL 的130 h。(4)褐云玛瑙蜗牛卵发育对1.0 g/mL 的4种入侵植物水提取液的干扰表现更为敏感。1.0 g/mL的薇甘菊水提取液抑制蜗牛卵发育的效果最好, 抑制率达81.67%,且与其它处理组间存在显著差异(P < 0.05)。而1.0 g/mL的马缨丹提取液优于五爪金龙提取液对褐云玛瑙蜗牛卵的抑制效果则表明入侵植物内源次生成分对不同发育时期的动物的防控效果存在差异。1.0 g/mL入侵植物水提取液各处理组对褐云玛瑙蜗牛卵发育抑制效果由强至弱的顺序为:薇甘菊 > 南美蟛蜞菊 > 马缨丹 > 五爪金龙。总之, 4种入侵植物对褐云玛瑙蜗牛成体、幼体和卵均具有较好的触杀与抑制作用, 利用入侵植物防控入侵动物, 以害制害, 化害为利, 是一种具有良好应用前景的环境友好型生物防治措施。     

褐云玛瑙螺在我国的适生性研究

研究了世界上重要的有害生物－褐云玛瑙螺在我国的适生性。结果表明,该螺的致互最低和最高温度,发育起点温度,世代有效积浊分别为：０．２,４１．２,１２．０℃和２４２２．６日度。在此基础上,综合应用越冬越夏界限,有效积温法则和生物相似距方法,用ＱＵＩＣＫ ＢＡＳＩＣ计算机语言建立了褐云玛瑙螺在我国的适生性区划系统模型。