Proposal for a new tapeworm order, Rhinebothriidea

The polyphyletic nature of the tapeworm order Tetraphyllidea Carus, 1863 is addressed in part with the establishment of the new order Rhinebothriidea for a subset of the taxa formerly comprising the phyllobothriid subfamily Rhinebothriinae (Platyhelminthes: Eucestoda). Support for the order comes from Bayesian, maximum likelihood, and parsimony analyses of complete ssrDNA and partial (D1-D3) lsrDNA sequence data for 58 cestode species. These data consisted of novel data generated for 40 species in 15 genera of candidate rhinebothriines and the cathetocephalidean species Sanguilevator yearsleyi as well as comparable data taken from GenBank for an additional 18 cestode species in 17 genera. In total, the species analyzed consisted of two Cathetocephalidea, two Litobothriidea, two Lecanicephalidea, three Proteocephalidea, and 49 Tetraphyllidea. The tetraphyllideans consisted of three Onchobothriidae, three Serendipidae, and 43 Phyllobothriidae (one Thysanocephalinae, one Echeneibothriinae, five Phyllobothriinae, 35 candidate Rhinebothriinae and the poorly known Spongiobothrium). This work suggests that some elements of current membership in the group are in need of revision. For example, while inclusion of the echeneibothriine genus Echeneibothrium and the phyllobothriine genera Rhodobothrium and Anthocephalum, and also Spongiobothrium, in the Rhinebothriidea is supported, inclusion of Duplicibothrium and Caulobothrium in the new order is not. Histological sections and scanning electron microscopy of selected members of the study group suggest that the presence of bothridial stalks may serve as an effective morphological feature to characterise the order. The group is restricted to elasmobranchs, and appears to have a particular affinity for Myliobatiformes. The new order includes at least 13 genera. Intraordinal relationships were determined to be insufficiently stable to justify the formal reorganization of rhinebothriidean families at this time.     

Chaperone characteristics of PDI-related protein A from Aspergillus niger

The functional properties of a novel protein, protein disulfide isomerase-related protein A (PRPA) from Aspergillus niger T21, have been characterized. (1) PRPA possesses disulfide isomerase activity. (2) In Hepes buffer, at substoichiometric concentrations, PRPA facilitates the formation of inactive lysozyme aggregates associated with PRPA (anti-chaperone activity); while at a high molar excess, PRPA inhibits aggregation by maintaining lysozyme in a soluble, yet inactive, state (chaperone-like activity). However, PRPA only exhibits chaperone-like activity during lysozyme refolding in phosphate buffer. (3) Experiments have indicated that disulfide cross-linkage is not required for the interaction between PRPA and lysozyme, and hydrophobic interaction may be responsible for PRPA effect on lysozyme. (4) Co-expression of PRPA and prochymosin in Escherichia coli leads to reduction of inclusion bodies, rendering part of prochymosin molecules soluble yet inactive. The structural and functional characteristics of PRPA suggest that PRPA may play an important role in protein folding, aggregation, and retention in the endoplasmic reticulum.     

Characteristics of the adaptation of the dwarf tapeworm to new hosts

Studies of the inbred lines of mice A/He, AKR and CBA infected with different strains of H. nana have shown that the helminth, when changing the host for another one of the same species but with different hereditary characters, happens to be insufficiently adapted to the new host. However it does not prevent the start of the infectious process. With increasing number of passages the parasite's adaptation level to the organism of the new host rises gradually. Possibilities and the adaptation level of the agent to the new host are defined as well by the adaptational mechanisms common to each specific strain of H. nana and the host's characters.     

Isolation and purification of a new protease from Aspergillus niger LCF no 9

Summary Aspergillus niger LCF N^o9 synthesizes large amounts of a semi-alkaline protease highly active on Benzoyl arginine p-nitroanilide (BAPNA) that is likely to find industrial use. An industrial-grade enzyme of around 80% purity was prepared and a reference enzyme was obtained in high yield from the industrial product by ultrafiltration and HPIEC on a mono Q column. The industrial-grade enzyme was produced on a pilot scale with a yield of 0.52% of medium solid starting material by adsorption on CM-Sephadex A₅₀, precipitation with acetone at –30°C, ion-exchange chromatography on DEAE-Sepharose and diafiltration.     

A new genus and species of proteocephalidean tapeworm (Cestoda) from Pangasius larnaudii (Siluriformes: Pangasiidae) in Southeast Asia

A new proteocephalidean cestode is described from spot pangasius, Pangasius larnaudii (Siluriformes: Pangasiidae), from Tonle Sap Lake in Cambodia and a new genus, Pangasiocestus , is proposed to accommodate it. The genus is placed in the Gangesiinae because its scolex possesses a large rostellum-like apical organ and its genital organs (testes, ovary, vitellarium, and uterus) are situated in the medulla, with some vitelline follicles paramuscular. Pangasiocestus romani n. gen. and n. sp., the type and only species of the new genus, is characterized mainly by its rosette-like scolex composed of 4 lobes bearing a small sucker in their center, and the apical part with a large, discoidal, rostellum-like apical organ devoid of hooks, by weakly developed inner longitudinal musculature formed by very few isolated muscle fibers, uneven size of testes in immature and mature proglottids, with lateral testes smaller and more dense than median ones, by very narrow lateral bands of vitelline follicles, formed usually by single follicles, and by the vagina anterior to the cirrus sac. This is the first proteocephalidean cestode from a pangasiid catfish identified to the species level (proteocephalidean cestodes from 3 Pangasius spp. reported in an unpublished account from Vietnam, misidentified as Proteocephalus osculatus (Goeze, 1782) [?= Glanitaenia osculata ], are not considered).     

A new affinity ligand for the isolation of a single 'feruloyl esterase' (FAE-III) from Aspergillus niger

8-Aminooctyl 5'-S-coniferyl-5'-deoxy-thio-alpha-L-arabinofuranoside has been synthesised and shown to be a selective affinity ligand for the feruloyl esterase III of Aspergillus niger. The hydrolyses of methyl 5-O-coumaroyl, feruloyl, or sinapoyl alpha-L-arabinofuranosides by this enzyme proceed at comparable rates.     

A novel NADPH/NADH-dependent aldehyde reduction enzyme isolated from the tapeworm Moniezia expansa

An aldehyde reduction enzyme has been purified from the cytosol of the tapeworm, Moniezia expansa, by chromatofocusing and Reactive-Red chromatography. The enzyme is monomeric (subunit 34 kDa) and can utilise NADH and NADPH as co-factors. Substrates of the enzyme include alkanals, alka-2,4-dienals and alk-2-enals, established secondary products of lipid peroxidation. The enzyme reduced methylglyoxal, another possible natural substrate (M. expansa lacks glyoxalase I activity). The parasite enzyme may help form a final line of defence against cytotoxic aldehydes arising from host immune initiated lipid peroxidation.     

Malformin A1 as a Mammalian Toxicant from Aspergillus niger

Culture conditions for guanosine production were studied with Bacillus subtilis MG–1 that exclusively accumulated guanosine. Of components investigated, KH₂PO₄, KC1, Fe⁺⁺, Mn⁺⁺, NH₄NO₃ and sodium glutamate have played important roles for guanosine production. The optimal concentrations in the culture medium were 2.0 g, 0.9 g, 7.5 mg, 7.5 mg, 20. 4 g and 6.0 g per liter, respectively.

In particular, the extremely minor concentration of Mn⁺⁺ 0.01 ppm completely repressed guanosine production although the cells grew sufficiently. Amino acids mixture was necessary for cell growth, but not essential for guanosine production.

Under these conditions, MG–1 accumulated 15 g of guanosine per liter in a weight yield of 18.8% of consumed sugar. However, a large amount of acetoin was also found as a byproduct.     

Galactofuranoic-oligomannose N-linked glycans of alpha-galactosidase A from Aspergillus niger.

Extracellular alpha-galactosidase A was purified from the culture filtrate of an over-producing strain of Aspergillus niger containing multiple copies of the encoding aglA gene under the control of the glucoamylase (glaA) promoter. Endoglycosidase digestion followed by SDS/PAGE, lectin and immunoblotting suggested that glycosylation accounted for approximately 25% of the molecular size of the purified protein. Monosaccharide analysis showed that this was composed of N-acetyl glucosamine, mannose and galactose. Mild acid hydrolysis, mild methanolysis, immunoblotting and exoglycosidase digestion indicated that the majority of the galactosyl component was in the furanoic conformation (beta-D-galactofuranose, Galf). At least 20 different N-linked oligosaccharides were fractionated by high-pH anion-exchange chromatography following release from the polypeptide by peptide-N-glycosidase F. The structures of these were subsequently determined by fast atom bombardment mass spectrometry to be a linear series of Hex(7-26)HexHA(c2). Indicating that oligosaccharides from GlcNA(c2)Man(7), increasing in molecular size up to GlcNA(c2)Man(24) were present. Each of these were additionally substituted with up to three beta-Galf residues. Linkage analysis confirmed the presence of mild acid labile terminal hexofuranose residues. These results show that filamentous fungi are capable of producing a heterogeneous mixture of high molecular-size N-linked glycans substituted with galactofuranoic residues, on a secreted glycoprotein.     

Cellulase Enzyme from Aspergillus niger

Cellulase enzyme was produced by a selected strain of Aspergillus niger isolated from deteriorated wood and grown on different carbon sources. Filter paper gave the highest yield, followed by carboxymethyl cellulose (CMC). Cellobiose as well as glucose gave a low yield, while the yield from lactose was negligible. The concentration of filter paper cellulose that induced the maximum yield of the enzyme was 1%. Both soluble cellulose (CMC) and cotton cellulose treated with phosphoric acid (swollen) were easily hydrolyzed by cellulase; an increase in cellulase concentration lead to more hydrolysis of CMC and gave linearity in the reaction velocity. At certain concentrations of the enzyme, increase in CMC concentration, (up to 1%) resulted in more reducing sugar. Beyond this point no more hydrolysis occur.