Double trouble for grasshopper molecular systematics: intra-individual heterogeneity of both mitochondrial 12S-valine-16S and nuclear internal transcribed spacer ribosomal DNA sequences in Hesperotettix viridis (Orthoptera: Acrididae)

Abstract.  Hesperotettix viridis grasshoppers (Orthoptera: Acrididae: Melanoplinae) exhibit intra-individual variation in both mitochondrial 12S-valine-16S and nuclear internal transcribed spacer (ITS) ribosomal DNA sequences. These findings violate core assumptions underlying DNA sequence data obtained via polymerase chain reaction (PCR) amplification for use in molecular systematics investigations. Undetected intra-individual variation of this sort can confound phylogenetic analyses at a range of taxonomic levels. The use of a DNA extraction protocol designed to enrich mitochondrial DNA as well as an initial long PCR of approximately 40% of the grasshopper mitochondrial genome failed to control for the presence of paralogous mitochondrial DNA-like sequences within individuals. These findings constitute the first demonstration of intra-individual heterogeneity in mitochondrial DNA-like sequences in the grasshopper subfamily, Melanoplinae, and only the second report of intra-individual variation in nuclear ITS ribosomal DNA sequences in grasshoppers. The fact that intra-individual variation was detected in two independent DNA marker sets in the same organism strengthens the notion that the orthology of PCR-derived DNA sequences should be examined thoroughly prior to their use in molecular phylogenetic analyses or as DNA barcodes.     

Fence-Line Contact Between Wild and Farmed Cervids in Colorado: Potential for Disease Transmission

Abstract: Direct and indirect contact between wild and farmed cervids along perimeter fences may play a role in transmission of diseases like chronic wasting disease (CWD), but no studies have quantified such interactions. At 9 high-fenced commercial elk (Cervus elaphus) farms in Colorado, USA, during October 2003 to January 2005, we used animal-activated video to estimate rates of fence-line use by wild cervids, rates of direct contact between farmed and wild cervids, and probability of direct contact when wild cervids were present. We recorded 8-fold-more wild elk per unit time than mule deer (Odocoileus hemionus) at fence lines. Depending on site, we recorded 0.66 to 46.90 wild elk per 1,000 hours of camera monitoring. We documented 77 interactions between wild and farmed elk involving naso-oral contact and no contact between wild mule deer and farmed elk. Rate of direct contact ranged from 0.00 to 1.92 direct contacts per 1,000 hours of camera monitoring among sites. Given recorded presence of wild elk, estimated probability of observing direct contact during a 2-minute video recording ranged from 0.00 to 0.11 among sites. Risk of direct contact was about 3.5 times greater for single woven-wire fence compared with offset electric fence attached to a single woven-wire fence. We observed no direct contact through double woven-wire fences. Because little is currently known about infection rates associated with infection mechanisms, we cannot infer a level of CWD infection risk from our results, but some form of double fencing should reduce potential for direct and indirect transmission of disease into or out of elk farms.     

A histochemical and electrophysiological study of the action of diazoxon on cholinesterase activity and nerve conduction in ganglia of the cockroach Periplaneta americana L

When sixth abdominal or metathoracic ganglia of the cockroach Periplaneta americana L. were irrigated continuously with diazoxon (O, O-diethyl O-(2-isopropyl-6-methyl-4-pyrimidinyl) phosphate) solution in situ, the log. of the time required to block conduction in certain nerve pathways in the ganglia was directly proportional to the log. of the concentration of diazoxon applied. Inhibition of cholinesterase began peripherally before function was affected, and had begun to affect the neuropile by the time that conduction was first blocked. Longer exposure to diazoxon disrupted nerve function even more, especially in the sixth abdominal ganglion, and inhibited more cholinesterase, but much longer exposure was needed to inhibit nearly all the cholinesterase. Irrigation with saline, begun when block first occurred, failed to restore completely either nerve function or cholinesterase activity. The cholinesterase activity of ganglia from cockroaches treated topically with an LD90 of diazoxon and examined at intervals after treatment decreased steadily to a level similar to that of ganglia treated directly with diazoxon until conduction was just blocked, but rarely became less, even in moribund insects. Nerve function in metathoracic ganglia became badly affected and remained so in all cockroaches that failed to recover, but sixth abdominal ganglia, though usually badly affected for a time, always recovered normal function, even in prostrate cockroaches. The condition of a poisoned insect, therefore, corresponded much more closely to the functional condition of the metathoracic ganglion than to that of the sixth abdominal ganglion. Applying the insecticide close to a ganglion advanced the time of onset of symptoms but affected the final outcome very little. It was calculated that the highest concentration of diazoxon in the haemo-lymph in contact with the nervous systems of cockroaches treated topically with LDgo's of diazoxon was about 10^-5M.     

Revision of Palaearctic mountain midges (Diptera: Deuterophlebiidae), with phylogenetic and biogeographic analyses of world species

Abstract. The mountain midges (Diptera: Deuterophlebiidae) of the Palaearctic Region are revised to include eight species. Four new species are described: D. brachyrhina sp. nov., D.oporina sp. nov. and D. blepharis sp.nov. from the Himalayas (Assam and Sikkim), and D.bicarinata sp. nov. from southern Korea. A lectotype is designated for D.mirabilis Edwards, and a key to adult males of all Palaearctic species is provided.
Larval, pupal and adult characters were used to reconstruct the phylogenetic and biogeographic relationships of world species of Deuterophlebiidae. Based on features of the adult male, the Himalayan species D. brachyrhina and D. oporina are considered the most primitive deuterophlebiids. The Nearctic species D. inyoensis is proposed as the sister group of the remaining species. Relationships among the latter are based primarily on larval and pupal characters; however, lack of information about the immature stages of several Palaearctic species contributes to a poorly resolved phylogeny. Several alternative hypotheses are presented and discussed. All phylogenetic alternatives suggest that the Nearctic fauna originated from at least two invasions of North America.     

Determining the site of brain tumors; the use of radioactive iodine and phosphorus

By tests using radioactive iodine combined with diiodofluorescein, the site of tumors was correctly determined in 61 per cent of 39 cases of tumors of the cerebral hemispheres. In 19 cases where the focal radioactivity was increased 24 per cent or more over that of the surrounding area, there were no errors in localization. Fifteen patients with expanding intracranial lesions were tested at operation with radioactive phosphorus and 14 lesions were correctly localized. This procedure in which the needle probe was used was found of great value in rapidly locating and outlining the area of involvement.