Partial sequence of hemoglobin from cobra (Naja naja naja)

Hemoglobin from the cobra snake, Naja naja naja, was isolated and its chains separated on a CM-cellulose column. The separation profile revealed an and two chains having the molar proportions of []₂,[ ₁]₁,[ ₂]₁. The N-terminal amino acid sequence of the intact chains and of the CNBr peptides were carried out. The ₂ chain was found to be heterogeneous comprising a minor component amounting to 11%. This later showed changes at two positions 9 and 14 in the first 30 residues sequenced.     

Cardiotoxin of the Indian cobra(Naja naja) is a pyrophosphatase

An inorganic pyrophosphatase has been purified to apparent homogeniety from Indian cobra(Naja naja) venom, with a ten-fold increase in specific activity. The enzyme activity is intrinsic to a protein fraction in the venom which is normally termed cardiotoxin, cobramine, cytotoxin and so on. The enzyme shows a lowK _m (70 μI) and high heat stability. The enzyme was active against sodium pyrophosphate; it also hydrolyses a few mononucletides and sugar phosphates at much lower rates. The physiological significance of inorganic pyrophosphatase in venom is discussed.     

Characterization of a cytotoxin-like basic protein from the cobra (Naja naja naja) venom

A cytotoxin-like basic protein has been isolated from the venom of the nominate race of cobra (Naja naja naja from Pakistan) by a single step of high-performance liquid chromatography. The primary structure was determined and consists of 62 amino acid residues in a single polypeptide chain. It is highly similar to that of the cytotoxin-like basic proteins isolated from other Naja species, but differs in two of the SS-loop structures from that of cytotoxins.     

Macrostylophora kinabaluae n. sp. (Insecta: Siphonaptera: Ceratophyllidae), a new flea from Sabah (Borneo, Eastern Malaysia)

This species, known only by a single male, is described from Mount Kinabalu and thus is recorded from the same area as Macrostylophora borneensis (Jordan, 1926), teste Traub (1972). It is distinguished from its congeneric and characterized by the absence of eriged setae on the thorax and first abdominal tergits, as well as by the shape of terminal segments and the phallosom. Macrostylophora kinabaluae was found to parasitize the Sciurid rodent Callosciurus prevostii that is widespread throughout most areas in the Malaysian subregion.     

Neutralization of cobra venom by cocktail antiserum against venom proteins of cobra (Naja naja naja)

Naja naja venom was characterized by its immunochemical properties and electrophoretic pattern which revealed eight protein bands (14 kDa, 24 kDa, 29 kDa, 45 kDa, 48 kDa, 65 kDa, 72 kDa and 99 kDa) by SDS-PAGE in reducing condition after staining with Coomassie Brilliant Blue. The results showed that Naja venom presented high lethal activity. Whole venom antiserum or individual venom protein antiserum (14 kDa, 29 kDa, 65 kDa, 72 kDa and 99 kDa) of venom could recognize N. naja venom by Western blotting and ELISA, and N. naja venom presented antibody titer when assayed by ELISA. The neutralization tests showed that the polyvalent antiserum neutralized lethal activities by both in vivo and in vitro studies using mice and Vero cells. The antiserum could neutralize the lethal activities in in-vivo and antivenom administered after injection of cobra venom through intraperitoneal route in mice. The cocktail antiserum also could neutralize the cytotoxic activities in Vero cell line by MTT and Neutral red assays. The results of the present study suggest that cocktail antiserum neutralizes the lethal activities in both in vitro and in vivo models using the antiserum against cobra venom and its individual venom proteins serum produced in rabbits.     

Acanthocephalus reunionensis n sp (Acanthocephala: Echinorhynchidae), a parasite of Anguilla species (Anguillidae) from Reunion Island

In a survey of 118 eels Anguilla bicolor, A. marmorata and A. mossambica, (Anguillidae) indigenous to Reunion Island in the Mascarene island group, western Indian Ocean, a new species of acanthocephalan, Acanthocepholus reunionensis n. sp., was found. With a proboscis hook formula of 19 rows of 4-5 hooks, and elongated cement glands arranged in three pairs, this species differs from all other species in the genus. This is the first record of the genus Acanthocephalus occurring in eels from the African Region.     

Preparation of highly potent Naja naja atra (Formosan cobra) antivenin.

As the preparation of high titer antiserum against Naja naja atra venom is a time-consuming process, attempts were made to develop a immunization procedure for producing highly potent antiserums within a short period. Rabbits were immunized for 12 weeks with (1) whole venom as used routinely in our Institute, (II) whole venom adsorbed on carboxymethyl-cellulose (CMC) and mixed with Freund's complete adjuvant and (III) neurotoxin adsorbed on CMC and adjuvant mixture followed by CMC-whole venom-adjuvant mixture. The results showed that one ml of the antiserum prepared by method (III) could neutralize 445 LD50 of whole venom, in other words, its potency was 4 and 40 times higher than those prepared by methods (II) and (I), respectively.     

Primary structure and functional properties of cobra (Naja naja naja) venom Kunitz-type trypsin inhibitor.

A trypsin inhibitor from the venom of the cobra Naja naja naja has been isolated by a single step of reverse-phase high-performance liquid chromatography. The protein strongly inhibits trypsin (Ki = 3.5 pM). The primary structure was determined by peptide analysis of the [14C]carboxymethylated inhibitor. The 57-residue polypeptide chain belongs to the family of Kunitz-type inhibitors, and exhibits 42% residue identity with bovine pancreatic trypsin inhibitor. The structure shows only 70% identity with the corresponding peptide from the Capa cobra (Naja nevia), establishing that the inhibitor molecule exhibits extensive variations. Functionally, a basic residue at position P3' correlates with strong inhibition.     

Caryospora cobrae n. sp. from the Indian Cobra Naja naja Linnaeus

Oocysts of Caryospora cobrae n. sp. were isolated from an Indian cobra, Naja naja Linnaeus from West Bengal, India. The sporulated oocysts were spherical to subspherical, 16.5-19.5 times 16.5-18.0 μm, and had a micropyle but lacked a polar granule and oocysl residuum. The sporocysts were piriform, measuring 12.0-16.5 times 9.0?12.8 μm; a Stieda body and sporocyst residuum were present.     

Cytoarchitectonic pattern of the hypothalamus in the cobra,Naja naja

Summary The distribution and cytoarchitectonic pattern of the magno- and parvocellular hypothalamic nuclei of the cobra, Naja naja, are described at the light-microscopic level. With respect to their tinctorial affinity to paraldehyde fuchsin (AF) as a representative of the Gomori-type of stains, the magnocellular neurons belong to the AF-positive and the parvocellular neurons to the AF-negative elements. In addition to the supraoptic and paraventricular nuclei proper, two accessory aggregations of magnocellular neurons, the nucleus retrochiasmaticus and nucleus circularis, can be identified. Although in a peculiar location, they may be regarded as subunits of the supraopticoparaventricular neurosecretory complex. As many as 22 AF-negative nuclear areas are identified in the hypothalamus of the cobra. The nucleus periventricularis hypothalami of earlier authors is subdivided into several circumscribed neuronal complexes. The nucleus arcuatus, nucleus hypothalamicus lateralis and nucleus lateralis recessus infundibuli are well developed. An attempt is made to interpret the significance of these nuclei on a comparative and phylogenetic basis.On leave from the Department of Zoology, Nagpur University, Nagpur, India     

Spiracanthus bovichthys n. gen. n. sp. (Acanthocephala: Arhythmacanthidae), a parasite of littoral fishes of the central south coast of Chile

A new acanthocephalan species, Spiracanthus bovichthys n. gen. n. sp., is described. Samples were taken from 26 marine fish species between 23 and 53 degrees S of Chile. The parasite was found in the intestine of 6 species and only between 36 and 40 degrees S, especially in those fish that prey on small crustaceans in the upper and sublittoral zone. The parasite was found in Bovichthys chilensis (Reagan). Auchenionchus variolosus (Valenciennes), Calliclinus genigutattus (Valenciennes), Sindoscopus australis (Fowler and Bean), Myxodes cristatus Valenciennes, and Gobiesox marmoratus (Jenyns). However, only the first species is a suitable host for the parasite as evidenced by the presence of mature females. This acanthocephalan belongs to Arhythmacanthidae, but it does not correspond to any genus described. Spiracanthus bovichthys is different in the number and spiralled distribution of its hooks in the proboscis. In the short proboscis, 3 groups of hooks are distinguished according to size, summing up to 150-190 hooks. The group of largest hooks are found in the apical part of the proboscis, and there are 10 diagonal rows of small hooks from the prebasal to basal proboscis. Its trunk is covered partially by small spines. Amphipod and isopods were the prey items shared among the host fish species and are the best candidates to be the intermediate hosts of S. bovichthys.     

Effect of modification of tyrosine residues in cytotoxin-1 from Indian cobra venom. (Naja naja naja)

The role of tyrosine residues in the biological activity of cytotoxin-1 was evaluated using N-bromo succinimide. N-bromo succinimide effected the oxidation of tyrosine residues in cytotoxin-1 with an increase in absorption at 260 nm. N-chloro succinimide was ineffective in the oxidation of tyrosine residues in the toxin. Oxidation of a single tyrosine residue (at 3.50 equivalents of N-bromo succinimide/mole of the toxin) resulted in complete loss of lethal activity of the toxin. The lytic activity of the toxin (lysis of erythrocytes) remained uneffected even after three of the four tyrosine residues in the toxin were oxidised.     

Isolation and characterization of a lethal phospholipase A2 (NN-IVb1-PLA2) from the Indian cobra (Naja naja naja) venom.

Indian cobra (Naja naja naja) venom is reported to contain multiple forms of phospholipase A2. Only a couple of them have been isolated and characterized. A lethal phospholipase A2 (NN-IVb1-PLA2) from Naja naja naja venom has been purified in three steps involving CM-Sephadex C-25, Sephadex G-50 and rechromatography on CM-Sephadex C-25 columns. It is a basic protein with pl value between 7-7.5 and has molecular weight between 11,000-11,500. The LD50 of NN-IVb1-PLA2 is 1.2 mg/K g body weight. It induces neurotoxic symptoms in the experimental mice and is devoid of myotoxic, anticoagulant, edema inducing and direct hemolytic activities.     

Variation in biochemical and pharmacological properties of Indian cobra (Naja naja naja) venom due to geographical distribution

Indian cobra (Naja naja naja) venom obtained from three different geographical regions was studied in terms of electrophoretic pattern, biochemical and pharmacological activities. SDS-PAGE banding pattern revealed significant variation in the protein constituents of the three regional venoms. The eastern venom showed highest indirect hemolysis and hyaluronidase activity. In contrast, western and southern venoms were rich in proteolytic activity. All the three regional venoms were devoid of p-tosyl-L-arginine methyl ester hydrolysing activity. The eastern venom was found to be most lethal among the three regional venoms. The lethal potency varied as eastern > western > southern regional venoms. In addition, all the three regional venoms showed marked variations in their ability to induce symptoms/signs of neurotoxicity, myotoxicity, edema and effect on plasma coagulation process. Polyclonal antiserum prepared against the venom of eastern region cross-reacted with both southern and western regional venoms, but varied in the extent of cross-reactivity by ouchterlony immunodiffusion and ELISA.     

Echinorhynchus brayi n. sp. (Acanthocephala: Echinorhynchidae) from Pachycara crassiceps (Roule) (Zoarcidae), a deep-sea fish

Echinorhynchus brayi n. sp. (Palaeacanthocephala: Echinorhynchidae) is described from Pachycara crassiceps (Roule) (Zoarcidae) from the Porcupine Seabight, Northeast Atlantic. The new species closely resembles E. canyonensis Huffman & Kliever, 1977, a parasite of a Pacific zoarcid, but has longer lemnisci, larger eggs and larger testes. E. brayi n. sp. can be readily differentiated from the ten other Echinorhynchus spp. recorded from deep-sea fishes (E. abyssicola, E. gadi, E. longiproboscis, E. malacocephali, E. melanoglaeae, E. muraenolepisi, E. petrotschenkoi, E. sebastolobi, E. trachyrinci and E. truttae), because it has fewer hooks per longitudinal row.     

Empruthotrema stenophallus n. sp. (Monogenea: Monocotylidae) from the nasal tissue of Dasyatis kuhlii (Dasyatidae) from Sabah, Borneo, Malaysia

Empruthotrema stenophallus n. sp. (Monogenea: Monocotylidae) is described from specimens from the nasal tissue of the blue-spotted maskray Dasyatis kuhlii (Muller and Henle, 1841) collected in shallow waters off Pulau Banggi and Pulau Mabul, Sabah, Borneo, Malaysia. This is the first monogenean species to be described from an elasmobranch collected from Sabah. E. stenophallus can be distinguished from the other 6 members of the genus by the morphology of the sclerotized male copulatory organ, which is narrow, short, and distally tapered. E. dasyatidis Whittington and Kearn, 1992, previously documented from the nasal tissue of several of elasmobranch species from Australia, is recorded from 8 host species distributed around Malaysian Borneo. These represent new host and locality records for this monocotylid. The difficulties in identifying species of Empruthotrema and the apparent lack of host specificity by some members of the genus are discussed.