Characterization of microRNAs expressed in the cystic legion of the liver of Mus musculus perorally infected with Echinococcus multilocularis Nemuro strain.

Alveolar echinococcosis (AE) is a zoonosis caused by the metacestode of Echinococcus multilocularis. The published genome of E. multilocularis showed that approximately 86% of its genome is non-coding. Micro RNAs (miRNAs) are small non-coding regulatory RNAs, and recent studies on parasitic helminths expect miRNAs as a promising target for drug development and diagnostic markers. Prior to this study, only a few studies reported the E. multilocularis miRNA profiles in the intermediate host. The primary objective of this study was to characterize miRNA profiles via small RNA-seq in E. multilocularis Nemuro strain, a laboratory strain of Asian genotype, using mice perorally infected with the parasite eggs. The data were then compared with two previously published small RNA-seq data. We identified 44 mature miRNAs as E. multilocularis origin out of the 68 mature miRNA sequences registered in the miRNA database miRbase. The highest quantities of miRNAs detected were miR-10-5p, followed by bantam-3p, let-7-5p, miR-61-3p, and miR-71-5p. The top two most abundant miRNAs (miR-10-5p and bantam-3p) accounted for approximately 80.9% of the total parasite miRNAs. The highly expressed miRNA repertoire is mostly comparable to that obtained from the previous experiment using secondary echinococcosis created by an intraperitoneal administration of metacestodes. A detailed characterization and functional annotations of these shared miRNAs will lead to a better understanding of parasitic dynamics, which could provide a basis for the development of novel diagnostic and treatment methods for AE.

Dembo polymerase chain reaction technique for detection of bovine abortion, diarrhea, and respiratory disease complex infectious agents in potential vectors and reservoirs

Bovine abortion, diarrhea, and respiratory disease complexes, caused by infectious agents, result in high and significant economic losses for the cattle industry. These pathogens are likely transmitted by various vectors and reservoirs including insects, birds, and rodents. However, experimental data supporting this possibility are scarce. We collected 117 samples and screened them for 44 bovine abortive, diarrheal, and respiratory disease complex pathogens by using Dembo polymerase chain reaction (PCR), which is based on TaqMan real-time PCR. Fifty-seven samples were positive for at least one pathogen, including bovine viral diarrhea virus, bovine enterovirus, Salmonella enterica ser. Dublin, Salmonella enterica ser. Typhimurium, and Neospora caninum; some samples were positive for multiple pathogens. Bovine viral diarrhea virus and bovine enterovirus were the most frequently detected pathogens, especially in flies, suggesting an important role of flies in the transmission of these viruses. Additionally, we detected the N. caninum genome from a cockroach sample for the first time. Our data suggest that insects (particularly flies), birds, and rodents are potential vectors and reservoirs of abortion, diarrhea, and respiratory infectious agents, and that they may transmit more than one pathogen at the same time.

Monitoring of environmental contamination by Echinococcus multilocularis in an urban fringe forest park in Hokkaido, Japan

<p><b>OBJECTIVES</b>The aim of this study was to determine the prevalence of Echinococcus multilocularis environmental contamination in an urban fringe-the Nopporo forest park of Sapporo city, Hokkaido, Japan. A secondary aim was to determine possible transmission risks areas by associating percentage occurrence of E. multilocularis-positive faeces with the different land-use classes.</p><p><b>METHODS</b>Wild fox faeces collected from the environment were examined by intravital methods, such as the taeniid egg sucrose floatation technique, E. multilocularis coproantigen enzyme-linked immunosorbent analysis and DNA test of taeniid eggs by PCR. Geospatial maps produced by the Global Positioning System and Landsat data were analysed using geographic information system software to determine the association between percentage occurrences of E. multilocularis-positive fox faeces and land-use classes.</p><p><b>RESULTS</b>Our findings showed high prevalence rates in both E. multilocularis egg and coproantigen-positive faeces (16 and 49%, respectively) in the investigated urban fringe forest park. Data revealed that percentage occurrence of E. multilocularis-positive fox faeces was associated with land-use classes, such as forest and open field (P < 0.05).</p><p><b>CONCLUSIONS</b>We conclude that Nopporo forest park in the urban fringe of Sapporo city, Hokkaido is a reservoir with a high prevalence of zoonotic infective agents for alveolar echinococcosis. Our findings suggest that interface habitats between forests or woodlands and open fields are indispensable for continued maintenance of the life-cycle of E. multilocularis and, as such, constitute high risk areas for echinococcosis transmission.</p>