RFLP proved to be a feasible auxiliary discrimination tool for selected Hyalomma species when access to sequencing methods is not available, as for instance during field studies. In addition, a polymerase chain reaction (PCR) based on the CO1 gene amplification of ticks has been developed for the unequivocal species identification by amplicon sequencing and specific restriction endonuclease cleavage pattern analysis. Moreover, MALDI TOF- MS was able to confirm morphologic findings where sequencing provided ambiguous results. In this study, six different Hyalomma species were tested, all of which were all clearly distinguishable by mass spectrometric analyses. For this purpose, a reference spectral library of several European, American and African tick species was established. Within the last years, MALDI-TOF MS analysis based on tick leg proteins has been shown to be a reliable method to distinguish several tick species. Therefore, we used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and restriction fragment length polymorphism (RFLP) analysis to distinguish the most common Hyalomma species from sub-Saharan Africa ( H. However, tick collection usually takes place far from analytical laboratories and therefore requires proper storage of samples. However, the morphologic species discrimination can be difficult for damaged or blood-fed ticks and in case of species intercrosses. MALDI-TOF MS has been used to identify tick species 2224 and to determine tick infectious status 2527. We also found that Ixori-PVK is expressed in other tick as well.The species identification of tick vectors of Crimean-Congo hemorrhagic fever virus (CCHFV), especially Hyalomma ( H.) species, is a prerequisite to understand the eco-epidemiology of this disease and to reveal vector and virus reservoir species. High energy collision-induced dissociation was successfully used to distinguish between Leu/Ile ambiguities in single cell preparations. Thus, the novel peptide may be involved in one of the key physiological processes in ticks. The function of this peptide in ticks is still unknown, but these pests consume large amounts of blood in a single blood meal which is a challenge for the regulation of diuretic processes. In mass spectrometry, matrix-assisted laser desorption/ionization (MALDI) is an ionization technique that uses a laser energy-absorbing matrix to create ions from large molecules with minimal fragmentation. The novel peptide (Ixori-PVK, PALIPFPRV-NH2) shows a high sequence homology with members of the insect periviscerokinin/CAP2b peptides which, in insects, are involved in the regulation of water balance. Technical Abstract: The first peptidergic neurohormone from ticks has been identified by using a combination of immunocytochemistry and mass spectrometric analysis of single cells. The work brings us one step closer to the development of practical neuropeptide-like substances that will be effective in controlling pest ticks in an environmentally friendly fashion. A deeper understanding of the specific structures of neuropeptides, where they are stored and released, and how these neuropeptides regulate critical processes in ticks will aid in the design of strategies to disrupt feeding and survival. The subject of this investigation is the Southern Cattle Fever tick, a pest of livestock in the US. We report on the use of state-of-the-art analytical techniques to unambiguously characterize the structure of the first regulatory neuropeptide of a species of tick via direct analysis of a single organ. New, selective control measures may be developed by designing metabolically stable mimics of these neuropeptides that actively inhibit or over-stimulate functions regulated by them, resulting in disruption of the internal environment of the insect or tick. The basic premise of this research is that neuropeptides (short chains of amino acids) serve as potent messengers in insects and ticks to regulate vital functions. Interpretive Summary: Because of problems with the development of resistance to conventional pesticides, there is a critical need for new concepts and alternative approaches in controlling tick pests. Biochemical and Biophysical Research Communications. To support the results of the MALDI-TOF MS tick species identification, 41tickspecimenscomprising3to5 specimens per tick species were used to create a reference spectra database, which was evaluated using the spectra of the 44 remaining tick specimens. Identification of tick periviscerokinin, the first neurohormone of Ixodidae: Single cell analysis by means of MALDI-TOF/TOF mass spectrometry. Submitted to: Biochemical and Biophysical Research CommunicationsĬitation: Neupert, S., Predel, R., Russell, W.K., Davey, R.B., Pietrantonio, P.V., Nachman, R.J.
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