ABSTRACT
The main goal of this study was to estimate the prevalence of Echinococcus granulosus among stray dogs, as well as its potential impact on the environmental contamination in the Kurdistan-Iraq using microscopic examination and the Copro-PCR method. The presence of taeniid eggs was recorded in 400 dog faeces collected from the four different regions in the Sulaimani Governorate. The parasite eggs were recovered from fresh and aged faecal samples of the dogs using two isolation techniques, a flotation method (Sheather's solution, modified; specific gravity: d = 1.27) and a sedimentation method (formal-ether) in which the sediments from dog faeces were collected. Both methods were used for Copro-PCR to detect the presence of Echinococcus species egg through DNA using common primers designed to amplify a partial gene of cytochrome oxidase subunit 1 (COX1). The results of the microscopic examination showed a higher prevalence rate, i.e., 97 (24.25%) of E. granulosus among stray dogs generally in Sulaimani Governorate. The prevalence of E. granulosus among stray dogs according to the district area was 40, 24, 23, and 20.8% in Rzgari, Kalar, Sulaimani, and Halabja, respectively. The positive samples (n = 50) were selected for molecular confirmation, the DNA was extracted from the sediment of the positive samples and 40 (80%) samples were successfully amplified by polymerase chain reaction. The sequences show that all samples belong to the Echinococcus granulosus sensu lato (G1-G3), with slight genetic variation. It was concluded that the sediment of dog faeces can be used for DNA extraction, which is a new method that increases the sensitivity of the test, and the amount of DNA yield would be higher than the routine method, which directly uses faeces of the dogs. In addition, the molecular diagnosis was more sensitive than the microscope examination for the presence of E. granulosus eggs. The prevalence of E. granulosus in both the final hosts and the intermediate hosts must be regularly monitored.
ABSTRACT
Room temperature iodocyclisation of homoallylamines stereoselectively delivers functionalised 2-(iodomethyl)azetidine derivatives in high yield. Increasing reaction temperature from 20 °C to 50 °C switches the reaction outcome to realise the stereoselective formation of functionalised 3-iodopyrrolidine derivatives. It was shown that these pyrrolidines are formed via thermal isomerisation of the aforementioned azetidines. Primary and secondary amines could be reacted with iodomethyl azetidine derivatives to deliver stable methylamino azetidine derivatives. With subtle changes to the reaction sequences homoallyl amines could be stereoselectively converted to either cis- or trans-substituted 3-amino pyrrolidine derivatives at will. The stereochemical divergent synthesis of cis and trans substituted pyrrolidines supports an ion part, aziridinium, isomerisation pathway for azetidine to pyrrolidine isomerisation. Six azetidine derivatives were probed in a zebrafish embryo developmental assay to detect potential biological effects through the analysis of morphology and motility behaviour phenotypes. The range of effects across the probed molecules demonstrates the suitability of this assay for screening azetidine derivatives. One of the probed molecules, rac-(((cis)-1-benzyl-4-phenylazetidin-2-yl)methyl)piperidine, exhibited particularly interesting effects in the developmental assay presenting with hypopigmentation and reduced circulation amongst others. This shows that the zebrafish embryo provides a fast, sensitive and effective way to screen new compounds and in the future in combination with existing in vivo and in vitro assays it will become an integral part in drug discovery and development.
