Functional and structural analysis of two fibrinogen-activating enzymes isolated from the venoms of Crotalus durissus terrificus and Crotalus durissus collilineatus

Fibrinogen-activating enzymes, widely distributed in Crotalidae and Viperidae venoms, are single-chain glycosylated serine proteases that display high macromolecular selectivity and are often referred to as thrombin-like enzymes （TLEs）. TLEs serve as structural models to extend our understanding of the structure-function relationships of blood coagulation factors, have been clinically used for the treatment of thrombotic diseases, and are used as tools in clinical assays. The combination of gel filtration and ion-exchange chromatography proved to be successful in obtaining milligram quantities of pure samples of TLEs from the venoms of Crotalus durissus terrificus （white venom） and Crotalus durissus collilineatus （yellow venom）. Functional characterization indicates that both enzymes preferentially degrade the Bβ chain of bovine fibrinogen and possess edema-inducing and coagulant activities. However, the TLE from C. d. collilineatus venom shows twofold higher coagulant activity with a minimum coagulant dose （MCD） of 0.6 μg/μl, whereas the enzyme isolated from C. d. terrificus indicated an MCD of 1.5 μg/μl. Molecular modeling of gyroxin and structural comparisons with other highly conserved snake venom serine proteases, underlines the key role played by the surface charge distribution and the double insertion in the 174surface loop in macromolecular substrate recognition by TLEs.     

Toxicological and biochemical characterizations of malathion sensitivity in two field populations of Oxya chinensis （Orthoptera： Acridoidea）

We evaluate comparative toxicity of malathion in the two populations of the grasshopper Oxya chinensis, collected from Daixian and Fanshi of Shanxi province, China. General esterases and acetylcholinesterase （ACHE） from the two populations were characterized and compared. LD50 of the Daixian population （7.58 μg/g body weight） was 2.02-fold higher than that of the Fanshi population （3.75μg/g body weight）. General esterase-specific activities in the Daixian population were 1.91,130 and 1.85-fold higher than those in the Fanshi population, when α-NA, α-NB and β-NA were used as a substrate, respectively. Kinetic studies of general esterase showed that Vmax values of general esterases hydrolyzing α-NA,α-NB and β-NA in the Daixian population were 2.15-, 1.12-, and 1.47-fold, respectively, higher than those in the Fanshi population. The AChE activity of the Fanshi population was 1.54-fold higher than that of the Daixian population. Kinetic analysis of AChE showed that significant differences were presented between the two populations in the Km values; and the Vmax value in the Fanshi population was higher than that in the Daixian population. Inhibition studies of AChE indicated that AChE from the Daixian population was 2.56-, 2.80-, and 2.29-fold less sensitive to inhibition by paraoxon, chlorpyrifos-oxon, and demeton-S-methyl, respectively, than that from the Fanshi population. These biochemical characterizations of general esterases and AChE were consistent with malathion bioassay in the two populations. It is inferred that the reduced sensitivity of altered AChE and increased general esterase activities play an important role in the differences of insusceptibility of Oxya chinensis to malathion between the two populations.     

Anti-oxidation activity of ethanol extracts from natural thalli of lichens

Screening test on anti-oxidation activity using 1,1-diphenyl-2-picrylhydrazyl(DPPH) was performed for 99 ethanol extracts of 85 species of natural thalli of lichens in order to find novel anti-oxidation compounds.The 17 extracts of natural thalli showed high anti-oxidation activity.Among them,the activities of extracts from Hypogymnia vittata,Peltigera aphthosa,Nephromopsis ornata,Pseudevernia furfuracea,Cladonia vulcani and Peltigera elizabethae were higher.Extracts of Peltigera spp.showed higher activity than those of other genera.The ethanol extract of P.aphthosa had been separated into ethyl acetate-soluble and water-soluble fractions.Two anti-oxidative spots were found only in the water-soluble fractions by thin-layer chromatography.The compound in the lower spot had the same Rf value,UV spectrum,and color as authentic solorinine that was previously found as a unique quaternary ammonium compound from Peltigera spp.We now report that the hydrophilic lichen substance,solorinine showed a nearly same anti-oxidation activity(EC50=120?mol/Lol/L) as standard antioxidant Trolox(EC50=150?mol/L).     

Differences in Thermal Preference and Tolerance among Three Phrynocephalus Lizards （Agamidae） with Different Body Sizes and Habitat Use

We acclimated adults of two viviparous(Phrynocephalus guinanensis and P.vlangalii) and one oviparous(P.versicolor) species of toad-headed lizards(Agamidae) to 28 °C,33 °C and 38 °C to examine whether thermal preference(preferred body temperature,Tp) and thermal tolerance(critical thermal minimum,CTMin;critical thermal maximum,CTMax) were affected by acclimation temperature,and correlate with body size and habitat use.Both Tp and CTMax were highest in P.versicolor and lowest in P.vlangalii,with P.guinanensis in between.The two viviparous species did not differ in CTMin and thermal tolerance range,and they both were more resistant to low temperatures and had a wider range of thermal tolerance than the oviparous species.Both CTMin and CTMax shifted upward as acclimation temperature increased in all the three species.Tp was higher in the lizards acclimated to 33 °C than in those to 28 °C or 38 °C.The range of thermal tolerance was wider in the lizards acclimated to 28 °C than in those to 33 °C or 38 °C.The data showed that:1) thermal preference and tolerance were affected by acclimation temperature,and differed among the three species of Phrynocephalus lizards with different body sizes and habitat uses;2) both Tp and CTMax were higher in the species exchanging heat more rapidly with the environment,and CTMin was higher in the species using warmer habitats during the active season;and 3) thermal preference and tolerance might correlat with body size and habitat use in Phrynocephalus lizards.     

Metabolic enzymes in seasonally acclimatized and cold acclimated rufous-collared sparrow inhabiting a Chilean Mediterranean environment

Due to the higher energy requirements of birds during winter, it is predicted that the activities of metabolic enzymes （e.g., citrate synthase, CS and cytochrome C oxidase, COX） should increase in tandem with increases in rates of energy expendi ture （e.g., basal metabolic rate, BMR）. However, there is mixed support for the hypothesis of enzymatic acclimatization. Fur thermore, there is little information about the effect of ambient temperature on energetics and tissue enzyme activity levels in passerines inhabiting seasonal Mediterranean environments. In this study we evaluated the interplay between BMR and enzyme activities of freshly caught individuals of the passerine Zonotrichia capensis in winter and summer in a Mediterranean environment from central Chile, and also in warm （30℃） and cold （15℃） labacclimated birds. The results revealed a lack of seasonal variation in BMR, thermal conductance and in the activity of CS and COX. However, we found higher BMR and lower thermal conductance in coldacclimated than in warmacclimated birds. Also, total CS activity was higher in the flight muscles of coldacclimated than in warmacclimated birds. We found also a significant correlation between BMR and total CS activity in pectoral muscle. Although some authors have suggested that BMR mainly depends on the metabolic intensity and mass of internal organs, our results revealed that skeletal muscles such as the flight muscles may also affect BMR. Finally, differences in the accli mation and acclimatization responses in Rufouscollared sparrows suggest that largescale oscillations in the physical environment might maintain the capacity for flexibility in thermogenic traits through generations .     

Energy intake,oxidative stress and antioxidant in mice during lactation

Reproduction is the highest energy demand period for small mammals, during which both energy intake and expenditure are increased to cope with elevated energy requirements of offspring growth and somatic protection. Oxidative stress life history theory proposed that reactive oxygen species(ROS) were produced in direct proportion to metabolic rate, resulting in oxidative stress and damage to macromolecules. In the present study, several markers of oxidative stress and antioxidants activities were examined in brain, liver, kidneys, skeletal muscle and small intestine in non-lactating(Non-Lac) and lactating(Lac) KM mice. Uncoupling protein(ucps) gene expression was examined in brain, liver and muscle. During peak lactation, gross energy intake was 254% higher in Lac mice than in Non-Lac mice. Levels of H2O2 of Lac mice were 17.7% higher in brain(P<0.05), but 21.1%(P<0.01) and 14.5%(P<0.05) lower in liver and small intestine than that of Non-Lac mice. Malonadialdehyde(MDA) levels of Lac mice were significantly higher in brain, but lower in liver, kidneys, muscle and small intestine than that of Non-Lac mice. Activity of glutathione peroxidase(GSH-PX) was significantly decreased in brain and liver in the Lac group compared with that in the Non-Lac group. Total antioxidant capacity(TAOC) activity of Lac mice was significantly higher in muscle, but lower in kidneys than Non-Lac mice. Ucp4 and ucp5 gene expression of brain was 394% and 577% higher in Lac mice than in Non-Lac mice. These findings suggest that KM mice show tissuedependent changes in both oxidative stress and antioxidants. Activities of antioxidants may be regulated physiologically in response to the elevated ROS production in several tissues during peak lactation. Regulations of brain ucp4 and ucp5 gene expression may be involved in the prevention of oxidative damage to the tissue.     

Seasonal changes of thermogenic capacity in Melano-bellied oriental voles (Eothenomys melanogaster)☆

Seasonal changes of thermogenic capacity in Melano-bellied oriental voles (Eothenomysmelanogaster) were studied by measurements of body mass, basal metabolic rate (BMR), non-shivering thermogenesis (NST), thermogenic properties of brown adipose tissue (BAT), relative weight of liver mass, mitochondrial protein (MP) content, and activities of mitochondrial cytochrome C oxidase (COX) in autumn (October), winter (December), spring (April) and summer (July), respectively.
Results showed that: (1) Body mass of E.melanogaster was the highest in autumn, and the lowest in summer. The seasonal change in body mass was subject not only by environment temperature but also to other factors such as reproduction, feeding habit and climate in the habitat. Reproductive periods of E.melanogaster were in early spring (February–March) and late autumn (September–October), and therefore, the body mass was higher in spring and autumn. In addition, E.melanogaster feeds on leaves and stems of plants in spring and summer, whereas mainly on fruits in autumn and winter as they contain more energy. The difference in the feeding habit could result in higher body mass in the late half of the year, but lower body mass in winter. The lower body mass in winter provides E.melanogaster with an advantage in motility. (2) The BMR and NST of E.melanogaster was higher in winter than in other seasons, which suggests that the regulating thermogenic capacity is an important adaptive strategy for E.melanogaster. However, there was no difference in relative oxygen consumption (NST–BMR/BMR), suggesting that the regulative capacity of BMR is approximately equal to NST in E.melanogaster. This is different from some small mammal species in north China, which have higher regulative capacity in NST than E.melanogaster. Melano-bellied oriental voles live in mountainous area with subtropical zone monsoon climate and abundant food resource, and therefore have lower regulative capacity in NST, which can reduce an absolute request for energy. The low regulative capacity in NST can also help E.melanogaster to maintain energy balance and stable body temperature. (3) The relative mass of BAT in winter was significantly higher than that in summer. Different seasons significantly affected the MP content of BAT. The MP content of BAT was in the following order: winter > fall > spring > summer. The trend as following was observed for the mitochondrial COX activity of BAT: winter > spring > fall > summer. In experimental conditions, the thermogenic rate of many small mammal species increased under cold acclimation. Short photoperiod alone could also induce E.melanogaster to increase NST and mitochondrial COX activity of BAT. Therefore, ambient temperature and photoperiod are important environmental cues to induce seasonal changes in thermogenic capacity in E.melanogaster. (4) The relative mass of liver in summer was higher than that in other seasons, but no seasonal changes were found among other seasons. The MP contents of liver showed highest level in winter and lowest in spring and summer. Significant seasonal changes were also observed in the mitochondrial COX activity of liver. It increased significantly in winter, but not in other seasons. This indicates that liver participate in adjustment of thermogenesis. These data indicate that both increasing whole-animal thermogenesis rate and thermogenic capacity at the cells of BAT and liver play an important role in the adaption of E.melanogaster to seasonal changes, and they are also closely correlated with the habitat condition and their life habit.     

Effects of elevated CO2 on the nutrient compositions and enzymes activities of Nilaparvata lugens nymphs fed on rice plants

Elevated CO 2 may reduce the tolerance of Nilaparvata lugen (N. lugens) to adverse environmental factors through the biological and physiological degeneration of N. lugens. In an artificial climate box, under 375 and 750 μL L 1 CO 2 levels, the rice stems nutrient content, the nutrient content and enzyme activities of N. lugens nymph fed on rice seedlings exposed to ambient and elevated CO 2 were studied. The results showed that rice stems had significantly higher protein and total amino acid levels under ambient than elevated CO 2 levels. Nymphs had significantly higher protein levels in the ambient CO 2 treatment, while their glucose levels were significantly lower under ambient CO 2 conditions. Significantly higher trypsin activity was observed in nymphs grown in elevated CO 2 . Significantly lower activities of the protective enzymes total superoxide dismutase and catalase were observed in the nymphs under ambient CO 2 . Meanwhile, the activity of the detoxification enzyme glutathione S-transferase was significantly higher in the ambient CO 2 treatment. Measuring how energy and resources were allocated to enzymes in N. lugens nymphs under elevated CO 2 conditions can provide a more meaningful evaluation of their metabolic tolerances to adverse climatic conditions.     

Validation of degree-day models for predict!ng the emergence of two fruit flies (Diptera: lephritidae) in northeast Egypt

We estimated thermal developmental thresholds(7o)and degree-day(DD)constants for the immature stages of two tephritid pests,Bcictrocera zonata(Saunders)and Ceratitis capitata(Weidenmann).Males of both species were trapped in an Egyptian guava orchard during the fruiting seasons of 2016 and 2017 and trap catches were compared with peak flights predicted by the DD model based on local weather data.Ceratitis capitata had faster development than B.zonata at 20 and 25°C,but their overall developmental rate was similar at 30 and 35°C.The thermal threshold of development(7o)of B.zonata was higher than that of C.capitata,indicating greater sensitivity to cold.Although 35°C yielded the fastest development of both species,survival was higher at 30°C,with B.zonata experiencing a slight advantage,suggesting better tropical adaptation.Immature development of B.zonata and C.capitata was estimated to require 338 and 373 d,respectively,and 616 and 424 DD for a complete generation.Trap catches over both seasons showed good correspondence to peaks of fly activity predicted by the DD models;deviations from expectation ranged from 0 to 7 d for both fly species.Both species had four overlapping generations per season,with B.zonata abundance peaking in the first generation in both years,but only in 2016 for C.capitata.The models predict about eight and 12 generations per year in northeast Egypt for B.zonata and C.capitata,respectively.These models should be useful for timing pest control measures to coincide with periods of peak fly activity in fruit orchards.     

Molecular differentiation of the microgastrine species commonly found in paddy fields from Southeast Asia,with additional data on their phylogeny （Hymenoptera：Braconidae）

Partial DNA sequences of three genes, that is, mitochondrial large ribosomal subunit (16S), nuclear large ribosomal subunit (28S D2) and mitochondrial NADH1 dehydrogenase (NADH1) gene, were sequenced from different microgas trine species(Braconidae: Microgastrinae) collected fresh from paddy fields. The DNA sequences were used to determine the extent of sequence variation among species in order to evaluate the specific status of each species. Cladistic analysis was also used to infer a phylogenetic relationship among these species. The results showed that sequence divergence among species of the same genus Cotesia was much lower than those among different genera, such as Cotesia, Exoryza and Apanteles; the sequence similarity of 16S rDNA and NADH 1 genes between Cotesia sp. and C. chilonis was higher than that between C. sp. and C. ruficrus.Phylogenetic analyses suggested that four species of Cotesia were always grouped in the same clade regardless of using different analysis methods; Cotesia sp. and C. chilonis are more closely related to each other than to C. ruficrus, different from previous morphological results. Additionally, sequence analyses indicated that NADH1 gene has more parsimony informative characters than 28S rDNA D2 and 16S rDNA at the species-level analysis,indicating that NADH1 gene might be a useful marker for species-level analysis.     

A comparative study of the biological properties of Dendroaspis (mamba) snake venoms.

1. The biological properties of twelve samples of venoms from all four species of Dendroaspis (mamba) were investigated. 2. Dendroaspis venoms generally exhibited very low levels of protease, phosphodiesterase and alkaline phosphomonoesterase; low to moderately low level of 5'-nucleotidase and very high hyaluronidase activities, but were devoid of L-amino acid oxidase, phospholipase A, acetylcholinesterase and arginine ester hydrolase activities. The unusual feature in venom enzyme content can be used to distinguish Dendroaspis venoms from other snake venoms. 3. All Dendroaspis venoms did not exhibit hemorrhagic or procoagulant activity. Some Dendroaspis venoms, however, exhibited strong anticoagulant activity. The intravenous median lethal dose of the venoms ranged from 0.5 microgram/g mouse to 4.2 micrograms/g mouse. 4. Venom biological activities are not very useful for the differentiation of the Dendroaspis species. The four Dendroaspis venoms, however, can be differentiated by their venom SDS-polyacrylamide gel electrophoretic patterns.     

A comparative study of the biological properties of some sea snake venoms.

1. The protease, phosphodiesterase, alkaline phosphomonoesterase, L-amino acid oxidase, acetylcholinesterase, phospholipase A, 5'-nucleotidase, hyaluronidase, arginine ester hydrolase, procoagulant, anticoagulant and hemorrhagic activities of ten samples of venoms from seven taxa of sea snakes were examined. 2. The results show that venoms of sea snakes of both subfamilies of Hydrophiinae and Laticaudinae are characterized by a very low level of enzymatic activities, except phospholipase A activity and, for some species, hyaluronidase activity. 3. Because of the low levels of enzymatic activities and the total lack of procoagulant and hemorrhagic activities, venom biological properties are not useful for the differentiation of species of sea snakes. Nevertheless, the unusually low levels of enzymatic activities of sea snake venoms may be used to distinguish sea snake venoms from other elapid or viperid venoms.     

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.     

A comparative study of the biological properties of Australian elapid venoms

1. The hemorrhagic, procoagulant, anticoagulant, protease, phosphodiesterase, alkaline phosphomonoesterase, L-amino acid oxidase, acetylcholinesterase, arginine ester hydrolase, phospholipase A, 5'-nucleotidase and hyaluronidase activities of 39 samples of venoms from 13 species (15 taxa) of Australian elapids were determined and the Sephadex G-75 gel filtration patterns for some of the venoms were also examined. 2. The results indicate that Australian elapid venoms can be divided into two groups: procoagulant Australian venoms (including N. scutatus, N. ater, O. scutellatus, O. microlepidotus, P. porphyriacus, T. carinatus, H. stephensii and P. textilis) and non-procoagulant Australian venoms (including A. superbus, P. colletti, P. australis, P. guttatus and A. antarcticus). 3. The non-procoagulant Australian venoms exhibited biological properties similar to other elapid venoms, while the procoagulant Australian venoms exhibited some properties characteristic of viperid venoms. 4. The data show that information on venom biological properties can be used for differentiation of many species of Australian elapids. 5. Particularly useful for this purpose are the hyaluronidase, alkaline phosphomonoesterase, acetylcholinesterase, and the procoagulant activities and the Sephadex G-75 gel filtration patterns of the venoms.     

乌梢蛇血清的抗出血因子:一个有前途的抗蛇毒药物原料

用柱层析和聚丙烯酰胺凝胶盘状电泳法,从乌梢蛇血清中分离纯化了一个抗出血因子。用SDS-聚丙烯酰胺凝胶电泳法测得其分子量大约为65 kD;测定了五种蝮亚科蛇毒(尖吻蝮、竹叶青蛇、原矛头蝮、哈扑和短尾蝮)的最小出血剂量和乌梢蛇血清中抗出血因子对这五种蛇毒的抗出血活性;还测定了七种蛇毒(除上述五种毒蛇外,还包括圆斑蝰和银环蛇)的半数致死量,以及抗出血因子对中毒小鼠的治疗作用。结果显示:从乌梢蛇血清中提纯的抗出血因子的抗蛇毒活性,不仅可以抵抗它的捕食者尖吻蝮的蛇毒,而且还可以抵抗具出血活性的其它蛇毒;但它对不具出血活性的银环蛇毒的致死抑制作用不明显。该抗出血因子不仅在体外实验表现出强的中和出血毒素的活性,而且在体内实验中亦表现出对中毒小鼠良好的治疗作用,因而可能成为新的抗蛇毒药物的有前途的原料。乌梢蛇血清对血循毒的中和能力的获得,可能归因于尖吻蝮与乌梢蛇之间捕食与被捕食相互作用的关系。     

Comparative characterisation of Russell's viper (Daboia/Vipera russelli) venoms from different regions of the Indian peninsula.

Russell's viper (Daboia/Vipera russelli) venom from different regions of India was subjected to chromatographic, electrophoretic, biochemical and immunological analysis. The elution profiles from ion-exchange chromatography and protein banding pattern from SDS-PAGE showed a significant variation in the constituents of venoms. The acidic proteins are found to be predominant in the venoms of eastern and western regions while basic proteins are the major contributors of the northern and southern regional venoms. The major variation of phospholipases A(2) in the venom samples of India may be described as: southern regional venom is rich in basic, toxic PLA(2) while this activity showed a dramatic decrease as one moves towards west, north and eastern regions of India. In addition, the caseinolytic, TAME-hydrolytic, anticoagulant, oedema-inducing and haemorrhagic activities of the venoms have also varied from one region to another. The muscle specimens of mice injected with venoms of different regions showed variable change in the muscle fibre damage and cell morphology. The eastern regional venom is most lethal among all the venoms. The lethal potencies for four regional venoms vary as: eastern>western>southern>northern. The polyclonal antibodies prepared against the venom of southern region showed cross-reaction with the venoms of other regions, but the extent of cross-reaction and diffusion patterns are different. However, the polyclonal antibodies prepared against southern regional venom showed no protection against lethal toxicity of other regional venoms.     

A comparative study of cobra (Naja) venom enzymes

1. The L-amino acid oxidase, hyaluronidase, alkaline phosphomonoesterase, protease, phosphodiesterase, acetylcholinesterase, phospholipase A and 5'-nucleotidase activities of 47 samples of venoms from all the six species of cobra (Naja), including five subspecies of Naja naja, were examined. 2. The results demonstrated interspecific differences in the venom contents of phospholipase A, acetylcholinesterase, hyaluronidase and phosphodiesterase. These differences in venom enzyme contents can be used for the differentiation of species of the genus Naja. 3. Thus, our results revealed a correlation between the enzyme composition of venom and the taxonomic status of the snake at the species level for the genus Naja.     

A comparative study of the biological properties of venoms of some old world vipers (subfamily viperinae).

1. The hemorrhagic, procoagulant, anticoagulant, phosphodiesterase, hyaluronidase, alkaline phosphomonoesterase, 5'-nucleotidase, arginine ester hydrolase, phospholipase A, L-amino acid oxidase and protease activities of 30 samples of venoms from nine species (12 taxa) of the old world vipers (Subfamily Viperinae) including snakes from the genera Bitis, Causus, Cerastes, Echis, Eristicophis and Pseudocerastes, were determined and the Sephadex G-75 gel filtration patterns for some of the venoms were also examined. 2. Examination of the biological properties of the venoms of the Viperinae tested indicates the presence of common venom biological characteristics at the various phylogenic levels. 3. Venoms of most species of the Viperinae examined exhibited characteristic biological properties at the species level, and this allows the differentiation of the Viperinae species by differences in their biological properties. 4. Particularly useful for this purpose, are the effects of venom on kaolin-cephalin clotting time of platelet poor rabbit plasma and the Sephadex G-75 gel filtration pattern and arginine ester hydrolase activity of the venom.     

A comparative study of the biological activities of venoms from snakes of the genus Agkistrodon (moccasins and copperheads)

1. The hemorrhagic, procoagulant, anticoagulant, phosphodiesterase, alkaline phosphomonoesterase, 5'-nucleotidase, hyaluronidase, arginine ester hydrolase, phospholipase A, L-amino acid oxidase and protease activities of 31 samples of venom from three species of Agkistrodon (A. bilineatus, A. contortrix and A. piscivorus) and 10 venom samples from five other related species belonging to the same tribe of Agkistrodontini were examined. 2. The results indicate that interspecific differences in certain biological activities of the Agkistrodon venoms are more marked than individual variations of the activities, and that these differences can be used for differentiation of the species. Particularly useful for this purpose are the phosphodiesterase, arginine ester hydrolase and anticoagulant activities of the venoms. 3. Venoms of the subspecies of A. contortrix and A. piscivorus do not differ significantly in their biological activities.     

Physiological and Biochemical Analysis of Factors in the Female Venom Gland and Larval Salivary Secretions of the Ectoparasitoid Wasp Eupelmus orientalis

The stinging adult female and the biting newly-hatched larva of the solitary ectoparasitoid wasp Eupelmus orientalis can both cause permanent paralysis and stop the development of Callosobruchus maculatus host larvae. These two processes of host envenomation appeared to be independent and complementary in primary parasitism or in hyperparasitism of a distantly related hymenopteran host species. In contrast, the development of larvae as hyperparasites on members of their own species or genus depended completely on the prior injection of female venom. The venoms of the female and the first instar larva had similar effects on the cellular metabolism of the primary hosts. Protein synthesis was blocked in C. maculatus hosts envenomated by a female or a first instar larva of E. orientalis, but the absence of DNA breakdown indicated that these paralysed hosts were alive and quiescent. The venomous secretions injected by adult females and first instar larvae of E. orientalis had distinct electrophoretic profiles. The immunoreactive features of proteins from female venom and larval secretions were also examined. There is evidence for antigenic conservation between some venom proteins of E. orientalis and Apis mellifera. Lastly, the hyaluronidase, phospholipase and lipase activities in the female venom gland and in larval-derived secretions of E. orientalis were assayed. No lipase activity was detected. Phospholipase activity was found in both the female venom and the larval secretions of E. orientalis, whereas hyaluronidase was specific to the female venom.