Purification of a novel antibacterial short peptide in earthworm Eisenia foetida

Earthworms live in an environment with abundantpathogens. These pathogens are, firstly, bacteria living inwater or soil that are ingested during feeding or introducedinto the body following injury. Parasites, particularlylarval forms, which represent the dissemination phase,are another important group of potentially pathogenicagents. During the course of evolution, earthworms havedeveloped defense strategies against these living patho-gens [1,2]. Earthworms lack true antibodies and hence anada…     

Reproductive potential of the earthworm Eisenia foetida

Summary Regression equations are provided for the earthworm Eisenia foetida with respect to age at which 50% of the population became clitellate at 25° C in relation to population density in activated sludge and in horse manure. Regression equations are provided for progeny per cocoon versus weight of cocoon, and weight of cocoon in relation to weight of parent; from these an equation is derived for progeny per cocoon relative to worm weight. Regression equations are given on (a) number of cocoons produced per adult in relation to age and population density from onset of adulthood to median peak production of cocoons, age 10 weeks, and from age 10 weeks to age 27 weeks, and (b) weight of worm in relation to population density and age between ages 5 and 27 weeks. From (a) and (b) a family of equations (c) are derived giving progeny per cocoon in relation to age of adult and population density. From equations (a) and (c) two families of equations are generated giving progeny per adult in relation to ascent to, and descent from, the median week of peak cocoon production in relation to population density. Data also are provided on age at which reproduction terminates in relation to population density, optimum population density for reproduction, and hatchability.     

Characteristics of cholinesterase of the earthworm Eisenia foetida

1. Except for pH optimum, reactivation and “aging”, the results indicate that the ChE of Eisenia foetida is distinct from acetylcholinesterase and butyrylcholinesterase.2. Propionylcholine was found to be a better substrate than acetylcholine, while butyrylcholine, benzoylcholine and acetyl-beta-methylcholine were found to be poor substrates for this ChE.3. Little or no tendency towards substrate inhibition was seen.4. The rate constants of inhibition by alkyl phosphates and carbamates favour the concept of this enzyme being a B-esterase, but different from the two main classes of ChE.5. The buffer-soluble and membrane-bound ChE showed no differences in substrate specificity and Michaelis-Menten constants.6. These enzymes separately also showed little or no tendency towards substrate inhibition.7. Crude homogenate, membrane-bound, buffer-soluble and Triton X-100 solubilized ChE, as well as fractions from isoelectric focusing are acted upon by inhibitors at rather similar rates.8. The presence of only one type of enzyme is therefore indicated.9. The existence of isoenzymes is not clearly indicated either by isoelectric focusing, by gel electrophoresis technique or density gradient centrifugation.10. The latter method indicated the molecular weight to be 108,000 ± 7000.     

Sites of expression of mRNA for lysenin, a protein isolated from the coelomic fluid of the earthworm Eisenia foetida

Lysenin is a 33-kDa protein of 297 amino acids that was originally purified from the coelomic fluid of the earthworm Eisenia foetida. It binds specifically to sphingomyelin. In this study, we attempted to identify the site of synthesis of lysenin in the earthworm. We detected the expression of mRNA for lysenin and the presence of immunoreactive lysenin in the large coelomocytes and in the free large chloragocytes present in the lumen of the typhlosole, a depression in the dorsal wall of the intestine. These coelomocytes and chloragocytes seemed to be mature and separate from the chloragogen tissue that lined the typhlosole. The free large chloragocytes in the typhlosole contained numerous vacuoles. The nuclei were small and irregular in shape, and glycogen granules and mitochondria were occasionally found between vacuoles. The chloragocytes of the chloragogen tissue that surrounded the coelomic side of the intestine and the dorsal blood vessel did not react with the lysenin antiserum and no expression of lysenin mRNA was detected in these cells. Furthermore, no evidence of the protein or of the mRNA was found in the cells of the pharyngeal gland. Our findings suggest that lysenin is produced in the free large chloragocytes in the lumen of the typhlosole.     

Nitrite biotransformation by mitochondria from the earthworm Eisenia foetida (Savigny).

1. Nitrite oxidase and cytochrome-c oxidase activity catalysed by cytochrome-aa3 were assayed in earthworms and rats. 2. Cytochrome-aa3 and intact mitochondria from the two species were anaerobically incubated in the presence of nitrite; the occurrence of mitochondria-induced nitrite biotransformations was evaluated by monitoring nitrite recovery in incubation medium. Possible nitric oxide production was also tested. 3. The ratio nitrite oxidase/cytochrome-c oxidase activity was much higher in earthworms than in rats. 4. Under anaerobic conditions and in the presence of respiratory substrates, earthworm mitochondria produced a time-dependent loss of nitrite in the incubation medium. On the contrary, rat mitochondria are unable to decrease environmental nitrite concentration. 5. Results support the notion that metabolic properties of earthworm mitochondria can be considered as an adaptation to chronic nitrite exposure, this toxicant being typically present in natural habitats of these worms.     

Examination of water balance and ionic regulation in the earthworm,Eisenia foetida

• 1.1. Weight change after submerging the earthworm into water varied remarkably according to the environmental humidity in which animals were placed before submergence.
• 2.2. Pretreatment with physiological saline solution before submergence in water gave stable values for the ionic concentrations of the body fluid.
• 3.3. Brain removal caused decrease of both sodium and chloride ion concentrations and increase of potassium ion concentration of the coelomic fluid when animals were submerged in water.
• 4.4. Although brain replacement failed, action of a brain hormone is suggested to regulate the decrease of both sodium and choride ions and increase of potassium ion of the coelomic fluid to normal level when animals were submerged in water.

     

Metabolic parameters of the earthworm Eisenia foetida in relation to temperature

Comparisons of rate of achieving sexual maturity, cocoon production, and growth rate at six temperatures between 15 and 30 degrees C led to the conclusion that the earthworm Eisenia foetida can be produced optimally over the range 18-25 degrees C and perhaps maximally at 20 degrees C.     

Immunochemical evidence for a gastrin-like peptide in the intestinal tissues of the earthworm Lumbricus terrestris

• 1.1. The presence of gastrin-like immunoreactive materials in intestinal tissues of the earthworm Lumbricus terrestris has been demonstrated by means of radioimmunoassay technique.
• 2.2. The greatest activity coincides with the areas of the intestines with high digestive enzyme activities. No gastrin-like activities were detected in nerve tissues.
• 3.3. The results also indicate that earthworm gut tissues may contain different forms of gastrin or gastrin-like peptides.
• 4.4. The absence of gastrin-like activity in earthworm nerve tissues may have significant phylogenetic implication for the proposed gastrin-cholecystokinin origin in neuronal elements of invertebrates.

     

Survival of body wall autographs, allografts, and xenografts in the earthworm Eisenia foetida.

It is acknowledged that autografts of Eisenia foetida body wall are accepted by their hosts. Allografts, however, have not been found to be accepted, although macroscopic observation of these might suggest they are, possibly owing to fusion of grafts and host epidermal cells. All xenografts eventually fail. Histological studies show that both allografts and xenografts were isolated from hosts, invaded, absorbed, and replaced with collagen. Statistical analyses of results from multiple transfers of xenografts show no evidence of hosts producing a heightened response of any nature to a second challenge of the same antigen.     

Development of a water hyacinth based vermireactor using an epigeic earthworm Eisenia foetida

The aim of this work was to investigate the potential of water hyacinth (WH) spiked with cow dung (CD) into vermicompost. Five vermireactors containing WH and CD in different ratios, were run under laboratory conditions for 147 days. The maximum worm growth was recorded in CD alone. Worms grew and reproduced favourably in 25% WH+75% CD feed mixture. Greater proportion of WH in feed mixture significantly affected the biomass gain, hatchling numbers and numbers of cocoons produced during experiments. In all the vermireactors, there was significant decrease in pH, TOC and C:N ratio, but increase in TKN, TK and TAP at the end. The heavy metals content in the vermicomposts was lower than initial feed mixtures. The results indicated that WH could be potentially useful as raw substrate in vermicomposting if mixed with up to 25% in cow dung (on dry weight basis).     

Expression and characterization of earthworm Eisenia foetida lumbrokinase-3 in Pichia pastoris

Lumbrokinase-3 (LK-3, AY438622), first cloned from the earthworm Eisenia foetida in our laboratory, is a component of earthworm fibrinolytic enzymes. In this study, cDNA encoding the LK-3 gene was sub-cloned into yeast pPIC9K expression vector and transformed into the Pichia pastoris GS115 cells by electroporation. High level expression of LK-3 in yeast cells was confirmed with a different induction time. The activity of expressed LK-3 was observed in fibrin plates. In addition, the expressed LK-3 protein could dissolve fibrinogen and bovine serum albumin. The use of this system for the high level production of biological protein is implicated from this study.     

Molecular cloning and characterization of cDNA encoding fibrinolytic enzyme-3 from earthworm Eisenia foetida

Earthworm fibrinolytic enzyme (EFE), a multi-com-ponent protease purified from some earthworm breeds,belongs to serine protease family with fibrinolytic activity[1]. It has been used in prevention and treatment of cardiacand cerebrovascular diseases in Ch…     

Purification and function of two analgesic and anti-inflammatory peptides from coelomic fluid of the earthworm,Eisenia foetida

The potential application of anti-inflammatory and analgesic compounds in medication and therapeutic care have become of increasing interest. We purified and characterized two novel analgesic and anti-inflammatory peptides, VQ-5 and AQ-5, from the coelomic fluid of the earthworm (Eisenia foetida). Their primary structures were determined as VSSVQ and AMADQ, respectively. Both peptides, especially AQ-5, exhibited analgesic activity in mouse models of persistent neuropathic pain and inflammation. AQ-5 also inhibited tumor necrosis factor alpha and cyclooxygenase-2 production. The mitogen-activated protein kinase signaling pathway, which is involved in analgesic and anti-inflammatory functions, was inhibited by AQ-5. Thus, the analgesic and anti-inflammatory effects of these peptides, especially AQ-5, demonstrated their potential as candidates for the development of novel analgesic medicines.     

The ultrastructural localisation of membrane-associated phosphatases in the integument of the earthworm Eisenia foetida

Summary Acid phosphatase was demonstrated in relation to the apical microvilli, though not in the epicuticular projections derived from them, and reaction products were rare at the level of the apical membrane between microvilli. Positive reactions occurred at the septate desmosomal level of the epidermal intercellular junctions. All reactions were associated with the inner leaflet of the membrane. The alkaline phosphatase technique was negative at all epidermal membrane sites.     

甲胺磷、乙草胺和铜单一与复合污染对蚯蚓的毒性效应研究

以赤子爱胜蚓(Eisenia foetida)为例,通过滤纸急性毒性试验法研究了东北黑土区普遍存在的3种农用化学品甲胺磷、乙草胺和Cu对蚯蚓的单一与复合毒性效应．单一实验结果表明,三者对蚯蚓均有毒性,它们的毒性顺序为乙草胺＞甲胺磷＞Cu,半致死浓度分别为：0．307、0．708和118．70mg.kg^-1;同时通过观察可知,乙草胺与Cu以皮肤渗入蚯蚓体的毒性效应较甲胺磷明显．复合毒性实验结果表明,低浓度Cu与高浓度Cu对甲胺磷的毒性均有增强作用;低浓度Cu对乙草胺的毒性有削弱作用,但高浓度Cu对乙草胺的毒性有增强作用．可见,三者对土壤生态系统生态安全性和土壤健康质量存在潜在危害,同时这几种污染物的共存进一步加大了潜在危害性,且复合毒性效应与各组浓度组合密切相关．     

9种农药对家蚕和蚯蚓的急性毒性

【背景】当前农药品种及其使用量日益增多,测试农药对环境生物的急性毒性成为农药环境安全监测的重要途径。【方法】采用食下毒叶法和药土法分别测定了甲氨基阿维菌素苯甲酸盐、阿维菌素、氰氟虫腙、螺螨酯、螺虫乙脂、嘧菌酯、苯醚甲环唑、戊唑醇、2,4-D二甲胺盐等9种农药对家蚕和蚯蚓的急性毒性,并根据其毒性等级划分标准进行分级,评价其对环境的安全性。【结果】甲氨基阿维菌素苯甲酸盐、阿维菌素、氰氟虫腙、螺螨酯、螺虫乙脂、嘧菌酯、苯醚甲环唑、戊唑醇和2,4-D二甲胺盐对家蚕的LC50(96 h)分别为2.05×10-3、8.59×10-4、2.79、250.48、11.52、272.18、2.50、1.93×10-2和534.47 a.i.mg·L-1,对蚯蚓的LC50(14 d)分别为11.05、6.29、100、100、100、100、99.13、115.31和100 a.i.mg·kg-1干土。其中,甲氨基阿维菌素苯甲酸盐、阿维菌素和戊唑醇对家蚕"剧毒",氰氟虫腙、螺虫乙脂和苯醚甲环唑对家蚕"高毒",其余农药对家蚕均为"低毒";阿维菌素对蚯蚓为"中毒",其余农药对蚯蚓均为"低毒"。【结论与意义】9种农药对家蚕和蚯蚓的急性毒性存在差异,为农药的合理使用和环境保护提供了依据。     

Analysis of oxygen equilibria of the giant hemoglobin from the earthworm Eisenia foetida using the Adair model

Oxygen equilibrium curves of the giant hemoglobin from the earthworm Eisenia foetida were determined at various concentrations of cations. Using the Adair model of 12 oxygenation steps, we succeeded in fitting the data better than the simple concerted model (MWC model). Analysis of the Adair constants (K1 to K12) indicated that the increase in oxygen affinity occurs in the last six steps (K7 to K12) of the oxygen binding and that it is enhanced by increase in Ca2+ concentration. The Hill coefficient (nmax) at pH 7.5 attained a maximum value of 9.76 at 20 mM CaCl2. In the presence of physiological levels of Ca2+ (5 mM), the Bohr effect was similar to that seen in vertebrates. The data were consistent with the release of two Bohr protons being accompanied by the oxygen-linked binding of one Ca2+. Mg2+ and Na+ exerted a similar effect on the hemoglobin, though to a lesser extent. The stoichiometry of Ca2+ binding of the hemoglobin revealed the presence of two classes of binding sites, of which the affinities are high (Ka = 8.8 x 10(3) +/- 103 M-1) and low. The number of high affinity sites per heme was found to be 0.3, comparable to the number of oxygen-linked Ca2+ binding sites.