EFFECTS OF OXYGEN AND ISOFLURANE ANESTHESIA ON HEMOLYMPH GAS ANALYSIS AND RIGHTING REFLEX OF ASIAN FOREST (HETEROMETRUS LONGIMANUS) AND DICTATOR SCORPIONS (PANDINUS DICTATOR).

Large arachnids are commonly managed under professional care, and anesthesia is occasionally required for physical examination and diagnostic and therapeutic procedures. Anesthetic responses and hemolymph gas analysis have been studied previously in spiders, but scorpions have yet to be investigated. This study measured hemolymph gas values with an i-STAT point of care blood gas analyzer in healthy adult Asian forest scorpions (Heterometrus longimanus = HL, n = 8) and dictator scorpions (Pandinus dictator = PD, n = 12) breathing: 1) room air (RA), 2) 100% oxygen for 10 min in a chamber (OX), and 3) 5% isoflurane and oxygen (ISO) in a chamber until induction or loss of righting reflex. All scorpions recovered without complications, and there were no cartridge failures. Analysis of hemolymph gas values revealed that pH was lower in OX compared with RA and ISO and was lower in PD compared with HL scorpions. The partial pressure of carbon dioxide did not differ between inhaled gases but was higher in PD compared with HL. The partial pressure of oxygen (pO2) was higher in ISO compared with OX, and both were higher than when breathing RA. Despite a lack of species difference in pO2, PD had a more dramatic increase in pO2 in ISO compared with HL (significant species and inhalant interaction). PD had a significantly shorter induction time than HL, but recovery time (return of righting reflex) did not differ between species. Subjectively, HL exhibited rough inductions compared with PD, characterized by violent whole-body and tail movements. The unexpected increase in pO2 in ISO compared with OX, along with the species-specific differences and anesthetic effects, emphasizes the unique respiratory physiology of scorpions and demonstrates that further species-specific studies of anesthetics are warranted.

Functionally redundant peg sensilla on the scorpion pecten.

All scorpions have two mid-ventral organs called pectines. Each pecten has thousands of pore-tipped sensilla sensitive to a variety of volatile organic and water-based stimulants. However, it was previously unknown whether individual sensilla were functionally identical or different. The information enhancement hypothesis predicts that all sensilla have similar chemosensitivities such that each is a unit of a parallel processing system. The information segmentation hypothesis states that sensilla differ in their chemosensitivities, a functional arrangement akin to the glomeruli-specific chemical detection system in the moth or human olfactory sense. In this study, we tested these hypotheses by extracellularly tip-recording sensillar responses to three aqueous tastants: 0.01 M KCl, 0.1 M citric acid, and 40% ethanol by volume. We isolated stimulation to one sensillum at a time and compared the chemoresponses. Sensilla appeared to respond similarly to the same stimulant (i.e., sensillar tip-recordings revealed activity of the same cell types), although sometimes a few sensilla responded with higher spike rates than the others. We conclude that our data primarily support the information enhancement hypothesis but for future tests of sensillar function we suggest a new hybrid model, which proposes that a few specialized sensilla exist among a mostly uniform field of identical sensilla.

Solution structure of BTK-2, a novel hK(v)1.1 inhibiting scorpion toxin, from the eastern Indian scorpion Mesobuthus tamulus.

The three dimensional structure of a 32 residue three disulfide scorpion toxin, BTK-2, from the Indian red scorpion Mesobuthus tamulus has been determined using isotope edited solution NMR methods. Samples for structural and electrophysiological studies were prepared using recombinant DNA methods. Electrophysiological studies show that the peptide is active against hK(v)1.1 channels. The structure of BTK-2 was determined using 373 distance restraints from NOE data, 66 dihedral angle restraints from NOE, chemical shift and scalar coupling data, 6 constraints based on disulfide linkages and 8 constraints based on hydrogen bonds. The root mean square deviation (r.m.s.d) about the averaged co-ordinates of the backbone (N, C(α), C') and all heavy atoms are 0.81 ± 0.23Å and 1.51 ± 0.29Å respectively. The backbone dihedral angles (ϕ and ψ) for all residues occupy the favorable and allowed regions of the Ramachandran map. The three dimensional structure of BTK-2 is composed of three well defined secondary structural regions that constitute the α-ß-ß structural motif. Comparisons between the structure of BTK-2 and other closely related scorpion toxins pointed towards distinct differences in surface properties that provide insights into the structure-function relationships among this important class of voltage-gated potassium channel inhibiting peptides.

Coevolution of diet and prey-specific venom activity supports the role of selection in snake venom evolution.

The processes that drive the evolution of snake venom variability, particularly the role of diet, have been a topic of intense recent research interest. Here, we test whether extensive variation in venom composition in the medically important viper genus Echis is associated with shifts in diet. Examination of stomach and hindgut contents revealed extreme variation between the major clades of Echis in the proportion of arthropod prey consumed. The toxicity (median lethal dose, LD(50)) of representative Echis venoms to a natural scorpion prey species was found to be strongly associated with the degree of arthropod feeding. Mapping the results onto a novel Echis phylogeny generated from nuclear and mitochondrial sequence data revealed two independent instances of coevolution of venom toxicity and diet. Unlike venom LD(50), the speed with which venoms incapacitated and killed scorpions was not associated with the degree of arthropod feeding. The prey-specific venom toxicity of arthropod-feeding Echis may thus be adaptive primarily by reducing venom expenditure. Overall, our results provide strong evidence that variation in snake venom composition results from adaptive evolution driven by natural selection for different diets, and underscores the need for a multi-faceted, integrative approach to the study of the causes of venom evolution.

Switch between tyrosinase and catecholoxidase activity of scorpion hemocyanin by allosteric effectors.

Phenoloxidases and hemocyanins have similar type 3 copper centers although they perform different functions. Hemocyanins are oxygen carriers, while phenoloxidases (tyrosinase/catecholoxidase) catalyze the initial step in melanin synthesis. Tyrosinases catalyze two subsequent reactions, whereas catecholoxidases catalyze only the second one. Recent results indicate that hemocyanins can also function as phenoloxidases and here we show for the first time that hemocyanin can be converted to phenoloxidase. Furthermore, its substrate specificity can be switched between catecholoxidase and tyrosinase activity depending on effectors such as hydroxymethyl-aminomethan (Tris) and Mg(2+)-ions. This demonstrates that substrate specificity is not caused by a chemical modification of the active site.

Domestic scorpion control with pyrethroid insecticides in Mexico.

Scorpion stings cause more morbidity in Mexico than any other country, leading to about 100 deaths annually. In 1999, the State of Morelos reported nine deaths among 30663 cases of scorpion sting. To replace lindane used for scorpion control, field trials of pyrethroid pesticides were undertaken in Morelos during 1998-2000 at the village of Chalcatzingo (population initially with 2760 inhabitants and 530 houses). Pre-intervention surveys detected scorpions (Scorpiones: Buthidae) of two species in the majority of houses: Centruroides limpidus limpidus Karsch outnumbering Vaejovis mexicanus smithi Koch. Scorpion prevalence was assessed, pre- and post-spray, directly by searching (40 min/house) and by householder reports of sightings inside houses. Pre-intervention perceptions of scorpion abundance were similar for all areas, with sightings in 12-18% of houses, whereas daytime searches detected more infestations in eastern parts of the village (prevalence 17% indoors, 22% outdoors) than in other sectors (9% indoors, 14% outdoors). Pyrethroids were evaluated as residual treatments in separate sectors of Chalcatzingo, with almost complete coverage indoors and peridomestically, using the following four formulations: bifenthrin 10% wettable powder (WP) applied at 50mg a.i./m2, cyfluthrin 10% WP (Solfac 10 WP) at 44-55 mg a.i./m2, deltamethrin 2.5% suspension concentrate (Biothrin 25 SC) at 11 mg a.i./m2 and 5% WP (K-Othrine 50 WP) at 35 mg a.i./m2. Phase 1 compared bifenthrin 10 WP, Solfac 10 WP and Biothrin 25 SC sprayed in December 1998; phase 2 compared Solfac 10 WP and K-Othrine 50 WP sprayed in June and again in December 2000, with follow-up surveys of scorpions one month post-spray and subsequently. Scorpion prevalence was reduced by 64-77% peridomestically one month post-spray and by 83, 46 and 15% in houses sprayed with cyfluthrin WP, bifenthrin WP or deltamethrin SC, respectively. Householder reports of sighting scorpions indoors were 33-85% below pre-intervention levels. Cumulative effects of the three spray-rounds over 3 years reduced scorpion prevalence by approximately 60% in the deltamethrin WP re-sprayed area and by approximately 90% in the cyfluthrin WP re-sprayed area. Householder sightings also fell by 67 and 28% in the cyfluthrin and delta-methrin re-sprayed areas, respectively. Operational efficacy of these products against scorpions at the dosages applied was ranked as cyfluthrin WP > bifenthrin WP > deltamethrin SC > WP. Reported cases of scorpion sting intoxication fell by 17% during this study after having risen by approximately 40% over four previous years.