A case of intestinal infarction following Vipera aspis bite

1. Cantharidin is a natural defensive toxicant produced by blister beetles. 2. Cantharidin shares structural similarity with highly toxic commercial herbicides (e.g., endothall, endothall anhydride and endothall thioanhydride). 3. Cantharidin inhibits the activity of purified catalytic subunits of serine/threonine protein phosphatases (PP) type 1 and type 2A. 4. Cantharidin increases force of contraction in isolated myocardial and vascular preparations. 5. Cantharidin enhances the phosphorylation state of myocardial and vascular regulatory proteins. 6. Cantharidin is a valuable tool for studying the function of PP in regulatory phosphorylation-dephosphorylation events.     

Purification and partial characterization of vitellin from the black-legged tick, Ixodes scapularis

Vitellin from the black-legged tick, Ixodes scapularis, was purified from eggs using gel filtration and ion exchange chromatography. The purified protein had a native molecular mass of 480 kDa. Under reducing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis; SDS-PAGE), vitellin was composed of seven polypeptides each at 154, 135, 87, 78, 67, 64 and 35 kDa. The isoelectric point was pH 6.9 and absorption maxima for the yolk protein were 280 and 400 nm. As in other ticks, vitellin from I. scapularis is also a hemoglycolipoprotein. Carbohydrates detected in vitellin were predominantly mannose with a small amount of N-acetylglucosamine. Lipids detected by thin layer chromatography (TLC) were triglycerides, free fatty acids, and cholesterol. Phospholipids associated with vitellin were phosphatidylethanolamine and phosphatidylcholine. Polyclonal serum produced in rabbits recognized vitellin from the eggs and ovaries, and vitellogenin from the hemolymph and fat body in reproductive females. This is the first report on the characterization of yolk proteins from a prostriate tick.     

Unusual three-dimensional bite mark evidence in a homicide case

A comparison has been made between bite marks observed on the nose of a female homicide victim and the configuration of the dental structures of a male suspect. The victim's bite marks showed a number of demonstrable characteristics which could be reproduced by models of the dentition of the suspect. Special peculiarities within and between individual teeth, together with the upper and lower jaw relationship, were demonstrably consistent with the bite mark patterns on the victim's nose. Detailed examination of the scientific evidence led the dental investigators to the conclusion that there was a positive match between the suspect's dentition and the bite mark on the decedent's nose. The dental investigation and subsequent dental testimony were prime factors leading to the defendant's conviction of manslaughter in the first case in California involving the major use of bite mark evidence.     

Isolation of a spotted fever group rickettsia from the Pacific Coast tick, Ixodes pacificus, in Oregon

A rickettsia of the spotted fever group was isolated on three occasions from Ixodes pacificus in western Oregon. These isolations, and additional evidence furnished by complement fixation tests on guinea pigs inoculated with other Oregon ticks of this species, indicate that the association of this rickettsia with the Pacific Coast tick may be widespread. This is the first isolation of a spotted fever group rickettsia from I. pacificus. Because the Oregon isolates are mildly virulent for guinea pigs they resemble the Western U and Rickettsia montana strains of rickettsiae. However, preliminary evidence from cross-immunofluorescence tests of mouse antisera suggests the Tillamook and Grants Pass strains are antigenically different from all known spotted fever group agents.     

Entomopathogenic nematodes as control agents of developmental stages of the black-legged tick, Ixodes scapularis

Thirteen species or strains of entomopathogenic nematodes of the genera Steinernema and Heterorhabditis were tested in vitro against unfed and engorged larvae, nymphs, and adults of the black-legged tick, Ixodes scapularis. Nematodes were pathogenic to engorged female ticks, but not to unfed or engorged larvae, nymphs, males, and unfed females. Steinernema riobravis (355) and Heterorhabditis megidis (M145) killed ticks most rapidly, with mean day of death postinfection of 2.5 and 3.5 days, respectively. However, all nematode strains and species were lethal to engorged female ticks within 7.5 days. Development of juveniles and reproduction of nematodes did not occur inside the tick cadavers, because no infective juveniles (IJs) emerged from cadavers after 40 days, and no nematode life-cycle stages other than the primary IJs could be isolated from cadavers. Egg masses were produced by 4 of 702 nematode-infected, engorged females. Egg masses produced were not significantly smaller than masses produced by control ticks; however, only 1 of the 4 egg masses hatched, whereas all of the egg masses produced by the control ticks hatched. Entomopathogenic nematodes may be useful as an alternative management method for I. scapularis populations, and may be more acceptable than acaricidal chemicals for use in infested areas.     

Electron microscopic study of Haller's organ in the tick, Ixodes persulcatus (Ixodidae)

Haller's organ consists of the capsule, in which are arranged 7 thin-walled porous olfactory hairs, and an anterior group of sensillae situated in the cuticular recess before the capsule. The anterior group is represented by one thick-walled porous, one conical, two thin and two grooved hairs. The porous hairs are characterized by "plugs", the grooved ones--by a complex system of cavities and tubules. Before the organ there are hairs resembling tactile and gustatory receptors of arthropods, behind the organ there arrange hairs with a system of cavities and tubules. Sensible elements of all sensillae studied are bipolar receptor cells bearing one modified cilium on the surface of peripheral processes. The details of the structure of perceptive and accessory apparatus of the above receptors were investigated.     

Immunization with a polyvalent OspA vaccine protects mice against Ixodes ricinus tick bites infected by Borrelia burgdorferi ss, Borrelia garinii and Borrelia afzelii

Sequence variability of the outer surface protein (Osp) A among Borrelia burgdorferi sl species suggests that a monovalent OspA vaccine may not protect against the various Borrelia present in Eurasia. Here, we confirmed that a monovalent recombinant OspA (rOspA) vaccine does not protect mice against Ixodes ricinus mediated infection with B. burgdorferi ss, Borrelia garinii and Borrelia afzelii. However, when mice were vaccinated with a cocktail of various rOspA from these three species, they were protected, and all challenge ticks that fed on them were cleared of their spirochetes. These results showed that a multiple OspA antigens vaccine, compatible with human use, was very efficient at protecting mice against B. burgdorferi ss, B. garinii, and B. afzelii.     

Characterization of Borrelia spp. isolated from the tick, Ixodes tanuki and small rodents in Japan

Rat pancreatic periacinar fibroblastoid cells (PFCs) appear to be involved in intralobular fibrosis and acinar cell regeneration. We isolated pancreatic acini of the rat, cultured the fibroblastoid cells, and characterized the cells morphologically and immunohistochemically. Isolated acini were seeded on culture dishes, and spindle-shaped cells migrated and proliferated. On Electronmicroscopic examination, microfilament bundles were seen, and the intracellular localization of vimentin, alpha-smooth muscle actin, and non-muscle myosin was identified immunohistochemically. These findings strongly suggest that the cells were myofibroblast-like. The PFCs were also demonstrated, immunohistochemically, to contain prolyl hydroxylase, type-I procollagen, type-III procollagen, type-IV collagen, fibronectin, and laminin. Stimulation by transforming growth factor beta 1 (TGF beta 1) increased intracellular immunoreactive prolyl hydroxylase and collagen synthesis in the PFCs. These findings indicate that PFCs proliferate in culture as myofibroblast-like cells and synthesize extracellular matrix components. It is possible that PFCs are involved in intralobular fibrosis in response to stimulation with TGF beta 1.     

A red-back spider bite in a lymphoedematous arm

We administered a record eight ampoules of antivenom over one week to a 55-year-old woman with a red-back spider bite on her chronically lymphoedematous right arm. She had severe local signs of envenomation but relatively little systemic effect. The final antivenom dose, injected subcutaneously at the bite site, had a good effect.     

Borrelia burgdorferi erp proteins are immunogenic in mammals infected by tick bite, and their synthesis is inducible in cultured bacteria

Borrelia burgdorferi, the causative agent of Lyme disease, can contain multiple genes encoding different members of the Erp lipoprotein family. Some arthropod-borne bacteria increase the synthesis of proteins required for transmission or mammalian infection when cultures are shifted from cool, ambient air temperature to a warmer, blood temperature. We found that all of the erp genes known to be encoded by infectious isolate B31 were differentially expressed in culture after a change in temperature, with greater amounts of message being produced by bacteria shifted from 23 to 35 degrees C than in those maintained at 23 degrees C. Mice infected with B31 by tick bite produced antibodies that recognized each of the Erp proteins within 4 weeks of infection, suggesting that the Erp proteins are produced by the bacteria during the early stages of mammalian infection and may play roles in transmission from ticks to mammals. Several of the B31 Erp proteins were also recognized by antibodies from patients with Lyme disease and may prove to be useful antigens for diagnostic testing or as components of a protective vaccine.     

Cytokine production by lymph node cells from mice infested with Ixodes ricinus ticks and the effect of tick salivary gland extracts on IL-2 production

In BALB/c mice repeatedly infested with nymphal Ixodes ricinus ticks, lymphocytes from axillary and brachial lymph nodes which drain the tick attachment site produced significant levels of IL-2, TNF-alpha and GM-CSF when stimulated in vitro with Con A or anti-CD3 antibodies. Cytokine production by cells from lymph nodes of the opposite flank was equivalent to that of cells from uninfested mice. Nine days after the first infestation and IL-2, GM-CSF were produced primarily by the CD4+ T cells, while some other cell types contributed also to the TNF-alpha production. In mice repeatedly infested, a gradual increase of lymph node cell production of IL-2 was observed. The IL-2 levels regularly increased from the first to the third infestation compared to TNF-alpha levels which gradually decreased. The in vitro production of GM-CSF was not affected by successive infestations. Spleen lymphocytes from naive mice produced higher levels of IL-2 than lymphocytes from axillary and brachial lymph nodes. Both tick salivary gland extracts and D-mannose inhibited IL-2 production by these lymphocytes.     

Duration of Borrelia burgdorferi infectivity in white-footed mice for the tick vector Ixodes scapularis under laboratory and field conditions in Ontario

The duration of Borrelia burgdorferi infectivity in white-footed mice (Peromyscus leucopus) experimentally inoculated or infested with infected Ixodes scapularis nymphs was evaluated. Infectivity was assessed by infesting these mice with unfed I. scapularis larvae at 7, 21, 35 and 49 days post-inoculation (DPI) or post-infestation (PI). At 7 DPI, B. burgdorferi was transmitted from 18 of 24 syringe-inoculated mice and all three tick-infected mice to I. scapularis larvae which fed upon them. However, at 21, 35 and 49 DPI, significantly fewer mice were infective. Borrelia burgdorferi was isolated from tissues of 14 of 22 syringe-inoculated mice about 56 DPI, and from all three tick-infected mice. However, the level of agreement between xenodiagnosis and bacterial culture was no greater than would be expected by chance alone. We also determined if B. burgdorferi infectivity of mice varied in relation to periods of tick feeding in the field. White-footed mice were trapped during April, July and August 1993 from two habitats on Long Point peninsula (Ontario, Canada), where B. burgdorferi is endemic. Mice from each habitat were infested with laboratory-reared I. scapularis larvae. Ticks from each mouse were subsequently examined by immunofluorescent assay for B. burgdorferi infection and mice were cultured for B. burgdorferi. None of 3577 I. scapularis larvae fed on 62 mice captured within the cottonwood dune habitat were infected with B. burgdorferi, although it was isolated from six of these mice. Within the maple forest habitat, 0/24, 8/21 (38%) and 1/21 (5%) mice transmitted B. burgdorferi to I. scapularis larvae during April, July and August, respectively. Most mice from the maple forest with B. burgdorferi-positive tissues (14/21) were collected during July, although the level of agreement between xenodiagnosis and tissue culture was poor. Because B. burgdorferi infectivity in mice appears to be of short duration, overwintered I. scapularis larvae and nymphs may have to feed upon infected hosts at the same time of year in order for a cycle of B. burgdorferi infection to be maintained on Long Point. Infected I. scapularis nymphs, rather than persistently infected vertebrate hosts, likely serve as the overwintering "reservoir" for B. burgdorferi on Long Point.     

Occlusion of the cervical internal carotid artery due to a dog bite wound: case report

A rare case of traumatic occlusion of the carotid artery at the neck caused by a dog bite is reported. A 10-year-old boy presented delayed onset of left hemiparesis after a dog-bite wound in the right neck. CT scan revealed a cerebral infarction in the right basal ganglia, and IV-DSA disclosed complete occlusion of the right internal carotid artery at the neck. The patient was treated conservatively and his left hemiparesis improved, but impaired visual acuity remained on the right side. Repeated IV-DSA 12 days later showed recanalization of the right internal carotid artery. The mechanisms of the traumatic occlusion of the internal carotid artery were discussed with reference to the literature.     

Seasonal variation in the role of grey squirrels as hosts of Ixodes ricinus, the tick vector of the Lyme disease spirochaete, in a British woodland

The partition of free fatty acids (FFA) to egg-phosphatidylcholine (egg-PC) and egg-phosphatidylethanolamine (egg-PE) vesicles was studied. Upon the addition of FFA to the suspension of vesicles, the pH of the aqueous phase changed depending on the length and saturation of the FFA hydrocarbon chain, as well as on the vesicle composition. The medium pH decreased faster if FFA was added to egg-PE as compared to egg-PC vesicles. The fluorescent free fatty acid indicator (ADIFAB) was used to measure the amount of FFA remaining in the aqueous phase. Most of the FFA added to the suspension of egg-PE vesicles remained in the aqueous phase, whereas in the presence of egg-PC vesicles the FFA partitioned preferentially into the lipid phase. The amount of FFA incorporated into the lipid bilayers was estimated by measuring the changes of pH at the lipid bilayer surface, using fluorescein-PE. At high surface concentrations of FFA, decreasing pH at the bilayer surface caused the protonation of FFA, and raised the pK of FFA at the bilayer surface from 5 to about 7. The partition of FFA in egg-PE vesicles was an order of magnitude lower than that in egg-PC vesicles. The incorporation amount was determined more by the molecular packing than by the nature of lipid headgroups, because steroylcaprioyl-PE, which preferred the bilayer structure, behaved more like egg-PC than egg-PE. Understanding FFA partition characteristics would help to interpret the hydrolysis measurements of phospholipids, and to explain many biological activities of FFA.     

Risk of contamination by Borrelia burgdorferi S. lato in a forest environment. Survey during 13 months of abundance of the tick Ixodes ricinus and its level of infestation by the Lyme borreliosis agent in Brittany

The authors, in a forest in Brittany previously studied for several years, caught by flagging, each month from April 1992 to May 1993, nymphs of I. ricinus tick, and looked by indirect immunofluorescence, for B. burgdorferi infestation. An amount of 1,506 ticks was thus studied. Infestation frequency was varying from 0 per cent in January and February to 14.4 per cent in August. Standarding of tick collecting method allowed to establish, for each month, a tick, "availability" index, and, according to the spirochete infestation frequency, to do estimation of the risk level, for human visiting the concerned forest, of being infected by B. burgdorferi. Obtained results show that this risk is the highest in August, and quite non-existent in January and February.     

Mark and recapture of adult Ixodes pacificus (Acari: Ixodidae) to determine the effect of repeated removal sampling on tick abundance

The effect of biweekly flagging on adult Ixodes pacificus Cooley & Kohls abundance was determined by using mark-recapture techniques. Ticks were recaptured up to three times during the 12-wk study, and increasingly higher proportions of marked ticks were recaptured as the study progressed. By week 6 of the study, 46% of the collected ticks had been marked, and on the final sampling date, all ticks had been captured previously. Removal sampling would have substantially reduced estimates of the abundance of the questing I. pacificus adult population. For ecological studies that require the repeated sampling of a given habitat, the most representative data are obtained by returning flagged ticks to their collection site.     

Identification of group VS116 strains among Borrelia burgdorferi sensu lato grown from the hard tick, Ixodes ricinus (Linnaeus, 1758) by off-coupled restriction fragment length polymorphism analysis

The degree of chemical shift similarity for homologous proteins has been determined from a chemical shift database of over 50 proteins representing a variety of families and folds, and spanning a wide range of sequence homologies. After sequence alignment, the similarity of the secondary chemical shifts of C alpha protons was examined as a function of amino acid sequence identity for 37 pairs of structurally homologous proteins. A correlation between sequence identity and secondary chemical shift rmsd was observed. Important insights are provided by examining the sequence identity of homologous proteins versus percentage of secondary chemical shifts that fall within 0.1 and 0.3 ppm thresholds. These results begin to establish practical guidelines for the extent of chemical shift similarity to expect among structurally homologous proteins.     

OspA antibodies inhibit the acquisition of Borrelia burgdorferi by Ixodes ticks

Ixodes ticks are infected by Borrelia burgdorferi when larvae feed on spirochete-infected mice. We studied the acquisition of B. burgdorferi by larval ticks, characterized the production of outer surface protein A (OspA) by spirochetes entering larvae, and examined the effects of OspA antibodies on the establishment of B. burgdorferi infections in ticks. Most larvae were infected by spirochetes 24 to 48 h after placement on mice. OspA antibodies stained the first spirochetes observed in larvae, suggesting that OspA is synthesized early during the colonization of the vector. When OspA antibodies were administered to B. burgdorferi-infected mice and larvae were then placed on the animals, the severity of larval infection and the number of infected ticks (7 of 16) were decreased compared with that of controls (15 of 16). The inhibitory effects of OspA antibodies were observed with passive antibody transfer as well as active host-generated immunity. The lower larval infection rate observed in the presence of OspA antibodies was exacerbated after the larval molt since only 1 of 12 nymphs was infected, and none of the mice that were fed upon by these nymphs became infected with B. burgdorferi. Therefore, an OspA antibody response in mice altered the reservoir competence of the vertebrate host by inhibiting the movement of B. burgdorferi from the host to the vector.