Mitochondrial Nad1 Research Articles

Factors contributing to organelle genomes size variation and the intracellular DNA transfer in Polygonaceae

The use of complete organelle genomes, including chloroplast and mitochondrial genomes, is a powerful molecular method for studying biological evolution and gene transfer. However, in the case of Polygonaceae, an important family with numerous edible, medicinal, and ornamental species, the mitochondrial genomes of only three species have been sequenced and analyzed. In this study, we present the mitochondrial and chloroplast genomes of two important Tibetan medicinal plants, Bistorta viviparum and B. macrophyllum. All the organelle genomes are assembled into a single circular structure and contain a common set of 32 protein-coding genes (PCGs). Some genes such as rps2 and ndhF were found to have high nucleotide polymorphism (Pi) in the chloroplast genomes, while cox1, mttB and rps12 showed pronounced Pi values in the mitochondrial genomes. Furthermore, our analysis revealed that most chloroplast genes and mitochondrial PCGs in Polygonaceae plants are under purifying selection. However, a few genes, including the chloroplast gene psaJ and the mitochondrial genes ccmFc, atp8 and nad4, showed positive selection in certain Polygonaceae plants, as indicated by a Ka/Ks ratio greater than one. Structural variation analysis revealed a wealth of differences between the mitochondrial genomes of five Polygonaceae species, with a particularly notable large-scale inversion observed between Reynoutria japonica and Fallopia aubertii. Furthermore, an analysis of the homologous sequences in the chloroplast and mitochondrial genomes revealed that the rps7 has been transferred from the chloroplast to the mitochondrial genome in all five Polygonaceae species. Finally, ecological niche models were constructed for B. viviparum and B. macrophyllum, indicating that mean annual temperature and altitude are the main climatic factors influencing the distribution of both species. Although the current distribution of B. viviparum is significantly wider than that of B. macrophyllum, projections suggest that the optimal growth ranges of both species will expand in the future, with B. macrophyllum potentially exceeding B. viviparum. This study not only contributes to the plastid genome database for Polygonaceae plants, but also provides theoretical insights into the adaptive evolution of these plants.

First molecular characterisation of Hydatigera taeniaeformis (Cestoda: Taeniidae) from Rattus rattus in urban market centres from Tamale, Ghana

BackgroundThe common cat tapeworm, Hydatigera taeniaeformis, is widely distributed across many regions of the world but its global genetic diversity, ecology, and other biological features are not fully understood. The predator-prey relationship of felids and rodents sustains the transmission in both sylvatic and synanthropic or urban settings. This study was carried out in three urban market centres in Tamale, Ghana to determine the prevalence of H. taeniaeformis infection in urban rats and characterise the local population genetics using the mitochondrial nad1 gene sequences.MethodsFrom January to May 2023, rats were captured and euthanized to identity and isolate the H. taeniaeformis metacestodes. The full mitochondrial nad1 gene was amplified, sequenced and used to characterise the cestode population.ResultsIn total 64/80 rats from the three market centres were found naturally infected with metacestodes of H. taeniaeformis. Averagely, infected rats harboured two cysts with one exceptional case of 25 cysts in one liver. Each cyst harboured one metacestode. The DNA sequences revealed four haplotypes in a star-like distribution network across the three market centres. All four haplotypes were found to be unique, clustered with H. taeniaeformis sensu stricto (s.s.) global isolates and had only marginal variation among themselves.ConclusionThis is the first account of the genetic variability within H. taeniaeformis in Ghana and would contribute to ongoing and future studies on African regional biogeographic distribution of the common cat tapeworm.

New species of Kontrimavichusia Makarikov & Binkienė, 2022 (Eucestoda: Hymenolepididae) from arvicoline rodents (Rodentia: Cricetidae) from the North Caucasus

Two previously unrecognized species attributable to the genus Kontrimavichusia Makarikov & Binkienė, 2022 in arvicoline rodents from the North Caucasus are described. Kontrimavichusia testiculata sp. nov. is described from Microtus majori (Thomas, 1906) from the northwestern Caucasus (Republic of Adygeya and Karachay-Cherkess Republic, Russia) and Kontrimavichusia hobergi sp. nov. is described from Microtus daghestanicus (Shidlovsky, 1919) from the central Caucasus (Republic of North Ossetia, Russia). Kontrimavichusia testiculata is readily distinguishable from K. asymmetrica (Janicki, 1904) and K. hobergi in having a larger number of testes (4–6 per proglottis), larger suckers and a longer cirrus and cirrus-sac. In addition, the new species differs from its congeners by the position of the cirrus-sac with regard to the poral osmoregulatory canals and position of distal end of the rostellar pouch relative to the posterior margins of the suckers. Kontrimavichusia hobergi can be readily distinguished from its congeners by the arrangement of the testes in a triangle and the position of the cirrus-sac with regard to the poral osmoregulatory canals. In addition, this previously unrecognized species differs from K. asymmetrica and K. testiculata by the smaller dimensions of the fully developed strobila and a narrower ovary. The cirrus-sac of K. hobergi is larger than that in K. asymmetrica but smaller than that in K. testiculata. We also used partial sequences of the nuclear ribosomal 28S rRNA gene and mitochondrial nad1 gen to justify the generic arrangement and independent status of these two new species which are characterized in the current manuscript.

Species identification of Panax ginseng throughout the entire industrial chain: From the ginseng field to highly processed products

Panax ginseng has been highly treasured in Asian countries due to its long history, more types of pharmaceutically active ginsenosides, and prominent pharmacological effects. However, the potential adulteration of P. ginseng with P. quinquefolius or P. notoginseng poses serious health risks to consumers and disrupts market stability within the ginseng industry. To address this challenge, a simple real-time PCR assay for rapid screening of field P. ginseng and a reliable multiplex PCR method for species identification of highly processed ginseng products were respectively established, based on the InDel markers derived from nuclear and mitochondrial intron sequences. The real-time PCR assay, coupled with a simple NaOH-Tris DNA extraction method, facilitates high-throughput screening of field ginseng samples based on the Tm values of PCR products with a single pair of primers. The multiplex PCR assay, targeting the mitochondrial nad1 and nad5 InDel markers, could detect exogenous adulteration of P. quinquefolius or P. notoginseng at a remarkable detection limit of 0.001 ng and successfully identified the original species in various forms of highly processed ginseng products. The results indicate that mitochondrial InDel markers resist DNA degradation better than nuclear ones, making them more suitable for species identification in highly processed ginseng products that contain trace amounts of degraded DNA. The present study provides robust InDel marker-based methods for species identification across the entire ginseng industrial chain, from the ginseng field to final processed products, thereby serving as reliable tools for ensuring the authenticity of P. ginseng products.

Molecular variation and phylogeography within European Isothecium alopecuroides and Pseudisothecium myosuroides (Bryophyta, Lembophyllaceae)

Abstract Earlier studies on Isothecium s.l. suggested potential interchange of genetic material between species that are nowadays known to belong to separate genera. In this study, we analyse the variation in three molecular markers (nuclear gpd; mitochondrial nad5; and plastid trnG) in a wide selection of Isothecium alopecuroides and Pseudisothecium myosuroides specimens to infer whether this potential exchange of genetic material is frequent. In addition, we explore the phylogeographic structure of these two species in Europe. Our results seem to show repeated genetic interchange for the nuclear and mitochondrial markers in both species, suggesting intergeneric hybridization, although horizontal gene transfer and/or incomplete lineage sorting could also explain the detected incongruences. The genetic lineages found indicate a complex biogeographical history for I. alopecuroides, including both post-glacial immigration from different glacial refugia and developing haplotypes potentially adapted to cold climates. As for P. myosuroides, genetic variants could either indicate a post-glacial immigration history or adaptation to oceanic conditions. Further studies including a larger set of more variable molecular markers could help to reach final conclusions on the results here presented.

Insights into U-to-C RNA editing from the lycophyte Phylloglossum drummondii.

The lycophyte Phylloglossum drummondii is the sole inhabitant of its genus in the Huperzioideae group and one of a small minority of plants which perform uridine to cytidine RNA editing. We assembled the P.drummondii chloroplast and mitochondrial genomes and used RNA sequence data to build a comprehensive profile of organellar RNA editing events. In addition to many C-to-U editing events in both organelles, we found just four U-to-C editing events in the mitochondrial transcripts cob, nad1, nad5 and rpl2. These events are conserved in related lycophytes in the genera Huperzia and Phlegmariurus. De novo transcriptomes for three of these lycophytes were assembled to search for putative U-to-C RNA editing enzymes. Four putative U-to-C editing factors could be matched to the four mitochondrial U-to-C editing sites. Due to the unusually few numbers of U-to-C RNA editing sites, P. drummondii and related lycophytes are useful models for studying this poorly understood mechanism.

Molecular phylogenetic analysis of Echinococcus granulosus sensu lato infecting sheep in Italy.

Cystic Echinococcosis (CE), caused by the larval form of Echinococcus granulosus sensu lato, is a neglected zoonosis still threatening public health worldwide. In Italy different epidemiological scenarios were reported depending on the geographical area and associated socio-economic activities. Although in northern Italy the occurrence of E. granulosus is considered sporadic, in the southern regions and, particularly in Sardinia, CE prevalence reaches high levels. We analysed CE cysts collected from infected sheep from various areas of mainland Italy and the Sardinia island, with the main objective to investigate intergenotypic and intragenotypic variations at national level. CE cysts were collected from slaughtered sheep following post mortem inspection at local abattoirs. Total genomic DNA was extracted and amplification and sequencing of the partial mitochondrial genes nad5 and cox1 were performed. A Bayesian phylogenetic tree was estimated on a nad5 dataset (n=260) composed of E. granulosus samples from this study (n=126) and all the nad5 haplotypes available in GenBank (n=134). In addition, haplotype network, diversity and neutrality analysis were performed on nad5 and cox1 sequences of Italian origin obtained in this study. E. granulosus sensu stricto (s.s.) was found to be the only Echinococcus species infecting sheep in Italy, mainly represented by G1 genotype (76%) and, to a lower extent, by G3 genotype (24%). Phylogenetic analyses revealed 40 nad5 and 33 cox 1 haplotypes, and the presence of two founder haplotypes, belonging to G1 and G3 genotype, showing 100% similarity with DNA sequences from different geographic regions. The lack of geographical segregation, high haplotype and low nucleotide diversity coupled with significant negative values of Tajima's D and Fu's Fs observed in this study indicated high genetic variation and demographic expansion of E. granulosus s.s. in Italy.

A New Record of Pogonatum tahitense (Polytrichaceae) from Tibet, China: Taxonomic Description, Range Expansion, and Biogeographic History

The genus Pogonatum stands out as the most diverse within the family Polytrichaceae, encompassing over 50 species. Pogonatum tahitense has been recorded across various Pacific regions, including Hawaii in the United States and Tahiti in French Polynesia, as well as in Asia, such as in Taiwan in China, Java in Indonesia, and Sabah in Malaysia. In the current study, a specimen collected in Tibet, China, is described, confirming its taxonomic classification as P. tahitense through a comprehensive analysis integrating morphological evidence and molecular study based on sequences from the plastid (rbcL, rps4, trnL-F), mitochondrial (nad5), and nuclear (ITS2) regions. This documentation represents the first record of the species within mainland China. A time-calibrated, molecular-based phylogenetic analysis was conducted, employing various approaches for ancestral range inference. The findings suggest that P. tahitense originated during the Pleistocene epoch, approximately 1.8 mya, in Tibet, China.

Cystic echinococcosis in cattle and sheep caused by Echinococcus granulosus sensu stricto genotypes G1 and G3 in the USA

BackgroundEndemic domestic dog-ruminant cycles and human cystic echinococcosis caused by Echinococcus granulosus have been sporadically reported in the United States. However, there is a paucity of molecular data describing the genotypes and haplotypes of this important cestode in domestic ruminant hosts.MethodsNinety-four cysts from the lungs and/or livers of slaughtered beef cattle (76 samples), dairy cows (five samples) and sheep (13 samples) were collected from abattoirs in four states of the USA. Samples were genotyped at two mitochondrial loci, cox1 and nad5. Sequences were used to determine species, genotypes and haplotypes using median joining networks and Bayesian phylogenetic analyses. Cyst fertility was assessed in hematoxylin and eosin-stained sections. Additionally, previously reported autochthonous E. granulosus infections in the USA in various hosts were mapped.ResultsBased on cox1 sequences obtained from 94 cysts, 89 (94.7%) were identified as E. granulosus G1/G3, while five (5.3%) were Taenia hydatigena. Taenia hydatigena were only isolated from sheep. Based on nad5 sequences obtained from 89 hydatid cysts, 96.6% and 3.4% belonged to E. granulosus sensu stricto genotypes G1 and G3 respectively. Two haplotypes were found among E. granulosus cox1 sequences, neither of which was geographically unique. Six haplotypes were found among nad5 sequences in genotype G1, of which five were novel, while one haplotype was found in genotype G3. In the concatenated cox1-nad5 dataset, seven haplotypes were identified, of which six were geographically unique. All cysts from cattle were non-fertile. Four cysts from sheep were fertile.ConclusionsAll genotyped samples belonged to E. granulosus s.s. This is the first study to our knowledge to confirm the presence of genotypes G1 and G3 in domestic cattle and sheep intermediate hosts in the USA and provide data for future diagnostic and epidemiological studies. Sequences have been deposited in GenBank (cox1 sequences: OR398494-OR398496, nad5 sequences: OR400695-OR400702).Graphical

Study on the genotypes of Echinococcus granulosus in yaks and sheep from Langkazi County in Tibet Autonomous Region of China based on mitochondrial cox1 and nad1.

To determine the genotypes of the epidemic strains of Echinococcus granulosus in livestock in Tibet, samples of E. granulosus cysts were collected from 11 yaks and 62 sheep. Genomic DNA was extracted from these samples, and gene fragments of mitochondrial cytochrome c oxidase subunit I (cox1) and NADH dehydrogenase subunit I (nad1) were amplified by PCR and sequenced. DNASTAR and MAGA7.0 were employed for homology analysis and phylogenetic tree construction. Echinococcus granulosus cysts were detected in 56.2% (41/73) of the samples screened. Of these, 63.4% (26/41) were identified as E. granulosus G1 genotype (common sheep strain), 24.4% (10 /41) as G3 genotype (buffalo strain), and 12.2% (5/41) were G6 genotype (camel strain). The study concludes that yaks and sheep in Langkazi county, Tibet, carry three E. granulosus genotypes (G1, G3, and G6), with the G1 genotype the predominant genotype in the region. This study clarifies the distribution of E. granulosus genotypes, providing genetic data and insight for the surveillance and prevention of echinococcosis.

A taxonomic revision of Hedwigia (Hedwigiaceae, Hedwigiales) in China

Hedwigia is a small genus that currently comprises only 12 accepted species. However, taxonomic issues within this genus remain not thoroughly resolved, and its species diversity in China is still debatable. In this study, we conducted an integrative taxonomic revision of Hedwigia in China based on both molecular and morphological evidence. We found an interesting species from Yunnan and Sichuan Province having an unknown combination of morphological characters. The molecular phylogenetic analyses based on mitochondrial nad5, chloroplast trnL-trnF and nuclear ITS markers suggest that this unknown moss represents a new species described here as H. sinica W.Z.Huang & R.L.Zhu, sp. nov. In addition, Hedwigia nemoralis Ignatova, Ignatov & Fedosov is proposed as a new synonym of H. filiformis (Michx.) P.Beauv. based on morphological and molecular evidence. Furthermore, H. czernyadjevae Ignatova, Ignatov & Fedosov, previously known only from Russia, is newly reported for China. The presence in China of H. stellata Hedenäs and H. ciliata (Hedw.) Boucher is doubtful because the voucher specimens of these two species were re-identified as H. sinica and H. emodica or H. filiformis, respectively. In conclusion, at present, four species of Hedwigia are confirmed in China.

Molecular and Immunological Characterization and Incidence Rate of Hydatid Cyst Isolated from Liver and Lungs for Human in Kurdistan Region, Iraq

Objective: This study aimed to identify characteristics, location, cyst stage, and species/genotypes of Echinococcus granulosus s.l. complex in humans from the Kurdistan region, Iraq. Surgical removal of 25 echinococcal cysts from 22 patients in Dohuk and Zakho cities (January 2020 to December 2022) was analyzed. Methods: Characteristics of 22 surgically confirmed cystic echinococcosis (CE) cases, including anatomical site, cyst fertility, and patientdemographics, were examined. Molecular analysis using mitochondrial NAD5 gene confirmed genotypes G1 or G3 of E. granulosus s.s.,indicating sheep–dog–human transmission. Results: Liver (78.6% fertile, 21.4% sterile) and lungs (81.8% fertile, 18.2% sterile) were the most common anatomical sites for CE in females (59.1%) compared to males (40.9%). All analyzed samples belonged to genotypes G1 or G3. No statistically significant correlation was found between genotypes and variables such as fertility, location, and cyst stage. Conclusion: The study underscores the prevalence of genotypes G1 or G3 in the E. granulosus s.l. complex, confirming sheep–dog–human transmission in Kurdistan, Iraq. Recommendations include monitoring and control programs in sheep and dog populations and public health education campaigns to reduce the risk of acquiring CE in humans.

Investigation and genetic polymorphism analysis of rodents infected with Echinococcus in Ili Prefecture, Xinjiang Uygur Autonomous Region, China.

Alveolar echinococcosis (AE) is a life-threatening disease in humans caused by the larval stage of Echinococcus multilocularis. Domestic animals, dogs, foxes, and small mammals constitute the circular chain of AE. To evaluate the infection, distribution, and genetic polymorphism of AE in the Ili Prefecture (Nilka, Xinyuan and Zhaosu), we conducted this survey. In June and July 2018, 267 small mammals were captured using water-infusion and mousetrap methods. Combined pathogenic and molecular biological methods were used to observe the histopathology of Echinococcus carried by rodents, amplify the mitochondrial nad1 gene of the pathogen, and investigate the genotype and haplotype diversity of Echinococcus in rodents in Ili Prefecture. Morphological identification revealed that these captured small mammals belonged to three species, with Microtus gregalis being the dominant species (183/267). Pathological and molecular biological results confirmed that E. multilocularis was the pathogen of echinococcosis in small mammals, with an infection rate of 15.73% (42/267). Among the three areas sampled, the highest infection rate of rodents was 25.45% (14/55) in Nilka County. However, there was no significant difference in the infection rates between regions (χ2 = 5.119, p > 0.05). Of the three captured rodent species, M. gregalis had the highest infection rate of 17.49% (32/183), but there was no significant difference in infection rates between the rodent species (χ2 = 1.364, p > 0.05). Phylogenetic analyses showed that the nad1 gene sequences obtained in this study clustered in the same clade as isolates from China. These isolates contained 21 haplotypes (Hap_1-21); Hap_2 was the most common haplotype (9/42). Furthermore, haplotype diversity (0.925 ± 0.027) and nucleotide diversity (0.01139 ± 0.00119) were higher in the Ili Prefecture than in other regions, indicating that population differentiation was high. Tajima's D and Fu's Fs tests were negative (p > 0.10), indicating that the population had expanded. The low fixation index (Fst) ranged from 0.00000 to 0.16945, indicating that the degree of genetic differentiation was different among different populations. In summary, Ili Prefecture is a high incidence area of AE, and Microtus spp. may play an important role in the transmission of AE in this area. The results of this study provide basic data for further study of the molecular epidemiology, genetic differences, and control of E. multilocularis in the Ili Prefecture, Xinjiang.

On the study of molecular markers of the genus <i>Picea</i> in Russia and Belarus

Currently, degradation of spruce stands is observed in various regions of Russia, which is associated with the transformation of climatic and environmental conditions of the environment. To ensure effective reforestation and afforestation of spruce forests, it is necessary to study the features of the population-genetic structure of plantations in order to preserve the genetic resources of valuable forest-forming tree species. The purpose of this work is to review modern achievements in the study of molecular markers of the genus Picea in Russia and Belarus. To characterize the genetic diversity of spruce populations, 9 ISSR primers with a high level of polymorphism (86,6%) were selected: (GA)9T, (AC)8C, (CA)6RY, (CA)6RG, (CA)6(GT), (CA)6(AC), (AG)8T, (GA)8C and (AG)8YT. The analysis of the mh44 locus of mitochondrial DNA revealed 13 allelic variants and is suitable for studying the genetic diversity of intra- and inter-population genetic variability of spruce. Experimental work with the mitochondrial Nad1 gene has shown the possibility of its application for phylogeographic studies of spruce. The assessment of the genetic variability of microsatellite loci Pt63718, Pt26081, Pt71936 revealed 19 allelic variants and is also suitable for phylogeographic studies. These results are very valuable and can form the basis for studying and characterizing the genetic structure of spruce plantations.

Genetic Analysis Based on Mitochondrial nad2 Gene Reveals a Recent Population Expansion of the Invasive Mussel, Mytella strigata, in China.

Mytella strigata is a highly adaptable invasive alien species that has been established in coastal China since 2014. Mitochondrial DNA (mtDNA) is an important tool for studying the evolution and population genetics of invasive species. In this study, the mitochondrial genome of M. strigata from China was sequenced by Illumina high-throughput sequencing and characterized with 13 protein-coding genes (PCGs). By assessing the selective pressure of 13 PCGs, the nad2 gene had the fastest evolutionary rate and was finally selected for population genetic analysis. A total of 285 nad2 sequences from seven M. strigata populations in China were analyzed and showed obviously T-rich and C-rich characteristics. According to population genetic diversity analysis, all the seven populations had haplotype (gene) diversity (Hd) ≥ 0.5 and nucleotide diversity (Pi) < 0.005. Haplotype networks showed a "star" distribution. Population historical dynamic analyses showed that Fu's Fs and Tajima's D values of all populations were negative except the Qukou (QK) and Beihai (BH) populations. The Zhangzhou (ZJ) and Xiamen (XM) populations were unimodal while the other populations were multimodal. These results suggested that the population of M. strigata in China may have passed the bottleneck period and is currently in a state of population expansion.

Detection of Asian genetic components in autochthonous human Echinococcus multilocularis infections from endemic Warmia-Masuria (north-eastern Poland)

Alveolar echinococcosis is a life-threatening zoonotic disease caused by the larval stage of the cestode Echinococcus multilocularis. People are aberrant intermediate hosts accidentally infected with the parasite eggs via faecal-oral route, usually by the consumption of unwashed fruit and vegetable or direct contact with definitive hosts. The recently reported presence of Asian admixture in E. multilocularis tapeworms from Polish red foxes prompted the question of metacestode descent in the human population. In this study, a Maximum Likelihood tree based on partial sequences of E. multilocularis mitochondrial genes cox1, cob, and nad2 coupled with a hierarchical clustering analysis of microsatellite EmsB profiles and supplemented by Sammon's nonlinear mapping with k-means clustering revealed Asian genetic components, to date associated only with the sylvatic cycle, in two autochthonous samples from alveolar echinococcosis patients living in endemic Warmia-Masuria, north-eastern Poland. The red fox is the most likely source of contamination in the environment shared by people and wildlife that led to these infections. Our results confirm that Asian genetic variants participate in the synanthropic cycle in north-eastern Poland and indicate that they may be present in the human population in other areas where Asian genetic variants were detected in red foxes.

Phylogeography and host‐shift speciation in the vampirecups, an enigmatic clade of endophytic holoparasitic plants

AbstractAimDiversification of obligate parasites is often closely associated with historical shifts among host species. This phenomenon is well‐documented in animal parasites but still poorly understood for parasitic plants. We studied the associations between host shifts, biogeography and diversification in Cytinus, a genus of endophytic holoparasitic plants which show a remarkable disjunct distribution.LocationMediterranean Basin, southern Africa, Madagascar.TaxonCytinus.MethodsWe sampled numerous populations in South Africa and the western Mediterranean Basin, as well as a few populations in Madagascar where the genus is rare. We amplified two nuclear (ITS and SSU) and two mitochondrial (matR and nad1) loci, to infer its evolutionary history using maximum likelihood and time‐calibrated Bayesian analyses, to elucidate the factors responsible for its current distribution and to assess whether host switching is a main driver of speciation.ResultsAll genes recover lineages corresponding to the three geographic areas of Cytinus and sublineages associated with specific host plants, although they disagree on their deep‐level relationships. Ribosomal nuclear genes support an initial split of the Malagasy lineage and an Afrotropical origin of Cytinus before colonization of warm temperate latitudes. Mitochondrial genes support an initial split of the Mediterranean and Afro‐Malagasy lineages and a later split between the South African and Malagasy clades. Introgressive hybridization is the most likely explanation for this discrepancy, while horizontal gene transfers from hosts is less likely. We found strong support for cryptic host‐specific diversity, with five and eight host races in the Mediterranean and South Africa respectively, and a strong phylogenetic signal in the host–parasite associations suggestive of parasite diversification via phylogenetic host tracking.Main ConclusionsThese dated phylogenies indicate that radiation through long‐distance dispersal events and adaptation to new hosts best explain the disjunct distribution of Cytinus. We emphasize the importance of host specificity and host‐switching in the diversification and biogeography of plant endoparasites.

When wildlife comes to town: interaction of sylvatic and domestic host animals in transmission of Echinococcus spp. in Namibia.

The present study was conducted in the isolated desert town of Oranjemund in the far south of Namibia. It is an extremely arid region where no livestock husbandry is practiced and only animals adapted to the desert can be found. However, in and around the city, artificial irrigation maintains lush green patches of grass that attract wild animals, in particular oryx antelopes (Oryx gazella). In 2015 four oryx antelopes were euthanised due to poor conditions and a post-mortem examination was conducted. Two were found positive for cystic echinococcosis and 16 cysts were collected for molecular analyses. In addition, faecal samples from black-backed jackals (n=5) and domestic dogs (n=9), which were regularly observed to feed on oryx carcasses, were collected and taeniid eggs isolated. Parasite species identification of the cysts and eggs was done by amplifying and sequencing the mitochondrial nad1 gene. Both oryx antelopes were found infected with E. ortleppi and one co-infected with E. canadensis G6/7. Both Echinococcus species were able to develop fertile cysts in oryx, making oryx antelopes competent hosts for these parasites. Therefore, the analysis of faecal samples was of high interest and although the numbers were quite small, taeniid eggs were found in three out of five faecal samples of jackals and in all nine dog samples. However, species determination was only successful with two jackal and one dog sample. All three were positive for E. canadensis G6/7. The absence of E. ortleppi may be due to the low number of faecal samples examined. In our small study, we discovered a rather unique lifecycle of Echinococcus spp. between jackals and domestic dogs as definitive hosts and oryx antelopes as intermediate hosts. Here, the presence of E. canadensis G6/7 is of particular concern, as it is the second most important causative agent of CE in humans.

Complete mitochondrial exploration of Echinococcus multilocularis from French alveolar echinococcosis patients

Alveolar echinococcosis (AE) is a parasitosis that is expanding worldwide, including in Europe. The development of genotypic markers is essential to follow its spatiotemporal evolution. Sequencing of the commonly used mitochondrial genes cob, cox1, and nad2 shows low discriminatory power, and analysis of the microsatellite marker EmsB does not allow nucleotide sequence analysis. We aimed to develop a new method for the genotyping of Echinococcus multilocularis based on whole mitochondrial genome (mitogenome) sequencing, to determine the genetic diversity among 30 human visceral samples from French patients, and compare this method with those currently in use. Sequencing of the whole mitochondrial genome was carried out after amplification by PCR, using one uniplex and two multiplex reactions to cover the 13,738 bp of the mitogenome, combined with Illumina technology. Thirty complete mitogenome sequences were obtained from AE lesions. One showed strong identity with Asian genotypes (99.98% identity) in a patient who had travelled to China. The other 29 mitogenomes could be differentiated into 13 haplotypes, showing higher haplotype and nucleotide diversity than when using the cob, cox1, and nad2 gene sequences alone. The mitochondrial genotyping data and EmsB profiles did not overlap, probably because one method uses the mitochondrial genome and the other the nuclear genome. The pairwise fixation index (Fst) value between individuals living inside and those living outside the endemic area was high (Fst = 0.222, P = 0.002). This is consistent with the hypothesis of an expansion from historical endemic areas to peripheral regions.

Update on the genetic diversity and population structure of Echinococcus granulosus in Gansu Province, Tibet Autonomous Region, and Xinjiang Uygur Autonomous Region, Western China, inferred from mitochondrial cox1, nad1, and nad5 sequences.

The identification of additional Echinococcus granulosus sensu lato (s.l.) complex species/genotypes in recent years raises the possibility that there might be more variation among this species in China than is currently understood. The aim of this study was to explore intra- and inter-species variation and population structure of Echinococcus species isolated from sheep in three areas of Western China. Of the isolates, 317, 322, and 326 were successfully amplified and sequenced for cox1, nad1, and nad5 genes, respectively. BLAST analysis revealed that the majority of the isolates were E. granulosus s.s., and using the cox1, nad1, and nad5 genes, respectively, 17, 14, and 11 isolates corresponded to Elodea canadensis (genotype G6/G7). In the three study areas, G1 genotypes were the most prevalent. There were 233 mutation sites along with 129 parsimony informative sites. A transition/transversion ratio of 7.5, 8, and 3.25, respectively, for cox1, nad1, and nad5 genes was obtained. Every mitochondrial gene had intraspecific variations, which were represented in a star-like network with a major haplotype with observable mutations from other distant and minor haplotypes. The Tajima's D value was significantly negative in all populations, indicating a substantial divergence from neutrality and supporting the demographic expansion of E. granulosus s.s. in the study areas. The phylogeny inferred by the maximum likelihood (ML) method using nucleotide sequences of cox1-nad1-nad5 further confirmed their identity. The nodes assigned to the G1, G3, and G6 clades as well as the reference sequences utilized had maximal posterior probability values (1.00). In conclusion, our study confirms the existence of a significant major haplotype of E. granulosus s.s. where G1 is the predominant genotype causing of CE in both livestock and humans in China.