Mitochondrial 16S rDNA Research Articles

Occurrence of autogeny in a population of Ornithodoros fonsecai (Acari: Argasidae)

Ornithodoros fonsecai is an argasid tick that is endemic to Brazil and has been described in the municipality of Bonito, state of Mato Grosso do Sul. Some specimens of this species were found in a cave in the municipality of Nobres, state of Mato Grosso. The specific identification of this population was confirmed by means of morphology and molecular biology. The mitochondrial 16S rDNA partial sequence of this species from Nobres has been deposited in GenBank (MK158949). The objective of this study was to elucidate the biology of O. fonsecai from Nobres, and to report autogeny in this tick population. Along three laboratory generations was observed molting of first nymphal instar to the second instar without feeding, a typical behavior of species included in the subgenus Alectorobius. The first generation (F1) presented five nymphal instars (N1 to N5), and most of adults emerged through molting of N5. The last nymphal instar of second generation (F2) was N4, but most of adults emerged from N3. In the third generation (F3) the last nymphal instar was N5, with most of the adults emerging from N4. In F2, some females (n = 20) originated from N3 began laying eggs without a blood meal. It was observed that those N3 fed twice before they molted to autogenic females. However, autogenic behavior occurred in relation to third generation females (F3) with specimens originating from N4 (n = 12) that were fed only once as nymphs. This behavior has already been reported as obligatory for the genera Otobius and Antricola, while it is facultative for one species of genus Argas and for four species of genus Ornithodoros. However, the present report provides the first record of facultative autogeny for a species of Ornithodoros in Brazil.

Prevalence and Molecular Characterization of Dirofilaria immitis (Filarioidea: Onchocercidae) in Dogs from Endemic Areas of Rio De Janeiro State, Brazil

Canine dirofilariasis is common in Brazil, but molecular diagnosis is rare even though molecular studies increase our knowledge about molecular epidemiology and circulating genotypes from helminths worldwide. This study aims to estimate the prevalence of infection with a modified Knott's test and to perform molecular characterization of Dirofilaria immitis (Leidy, 1856) Railliet and Henry, 1911, in dogs from endemic areas of Maricá and Niterói municipalities, Rio de Janeiro State, Brazil. Molecular characterization was performed in 33 blood samples from dogs positive for microfilariae and 4 adult worms obtained from 2 other dogs. DNA extraction followed by PCR for mitochondrial target 12S rDNA and cytochrome oxidase subunit 1 (COI) of D. immitis were performed, and the amplified products were sequenced. All sequences were identical for both gene targets and showed 100% identity with D. immitis sequences from different animal species from various countries. The study concluded that this genotype of D. immitis might be dispersed worldwide.

Severe coenurosis caused by larvae of Taenia serialis in an olive baboon (Papio anubis) in Benin.

In March 2017, a captive male juvenile (ca. 6 months old) olive baboon (Papio anubis) was brought to a primate rescue center in Benin with multiple subcutaneous swellings of unknown aetiology. At the general inspection of the body, around 15 partially mobile masses of variable sizes were found in different locations across the body. Following two surgical procedures, several cyst-like structures were removed and placed either in 10% formalin or in absolute ethanol. The cysts had a typical coenurus-like morphology. Genomic DNA was extracted from one cyst using a commercially available kit. The molecular characterization was performed by PCR amplification and sequencing of a region of the nuclear ITS-2 rDNA and a fragment of the mitochondrial 12S rDNA gene, revealing its identity as T. serialis, with 88%–98% similarity to T. serialis isolates from USA, Japan and Germany This study reports a new host for the larval form of T. serialis and confirms the role of primates as intermediate host, highlighting also the risk for human infection. Further genetic studies from multiple geographic isolates are needed to clarify the taxonomic status of this group.

Mitochondrial 16s rDNA profiling and phylogenetic analysis suggest genetic diversity of ash weevil (Stereonichus fraxini De Geer) in Serbia

This study contributes to knowledge of ash weevil (Stereonychus fraxini De Geer) molecular taxonomy, phylogeny and genetic diversity. Adult and larvae stages of insect were collected from several locations covering northern and central part of Serbia cojoined with homologous sequences with respect to their different geographic origin and hypothesis of their evolutionary relationships. Due to its slow rates of evolution the gene region that covers mitochondrial 16S rDNA, was choice for sequence profiling and phylogenetic reconstruction of ash weevil in correspondence with sequences of related tribes Cionini. Phylogenetic analyses demonstrating clear separation of the native weevil populations and the Cionini tribes. Even though bioinformatic tools confirm that all native specimens belong to species Stereonychus fraxini, different profile of the mitochondrial 16S rDNA in the clade of Serbian specimens indicate intraspecific genomic rearrangement in one specimen detached it to northern geographic position. Those particular specimens invade also different Fraxinus species. Genetic distinctness of other imagos from this particular individual proved by indels and point mutations found in their sequences. By screening the mitochondrial 16S rDNA, molecular evidence suggests the existence of the specimen with rearranged genome that indicate genetic variability in native populations of ash weevil species.

Molecular insights of mitochondrial 16S rDNA genes of the native honey bees subspecies Apis mellifera carnica and Apis mellifera jementica (Hymenoptera: Apidae) in Saudi Arabia

The honey bee Apis mellifera is of major importance for the world’s agriculture and is also suitable for environmental monitoring. It includes several recognized subspecies distinguished by using morphological and morphometric variants. Here, 200 adult worker Apis mellifera honey bees were collected from Hail region, Saudi Arabia. Mitochondrial 16S rDNA was conducted to detect molecular polymorphism among honey bee A. mellifera subspecies. The amplified and sequenced gene regions of mtDNA revealed the presence of two different subspecies of Apis mellifera carnica (gb| MH939276.1) and Apis mellifera jementica (gb| MH939277.1). The sequences were compared with each other and with others retrieved from the GenBank demonstrating a high degree of similarity (up to 72%). The NJ tree indicated that all Apis species are clustered together in one clade in addition to the genetically origin of Apis species within family Apidae as a paraphyletic group within the African lineage.

High level of genetic variation in mitochondrial 16S rDNA among populations of Porcellionides pruinosus (Brandt, 1833) (Crustacea: Isopoda: Oniscidea) in Tunisia

For more than 20 years, the cosmopolitan species Porcellionides pruinosus (Brandt, 1833) has been characterised by a controversial taxonomic status. This “species” has a high ecological plasticity, which enabled it to conquer different kinds of habitats. Besides that, this “species” exhibits a geographical variation in its morphological features and its reproduction pattern. In fact, some Tunisian populations had a seasonal reproductive period and while others showed a reproductive activity. A genetic analysis of rDNA 16S sequences has been performed to compare eight populations of P. pruinosus from different geographical sites, located in the north, centre, and south of Tunisia. Results reveal that the population of Tunis shows considerable variation in the primary structure of the sequences and has a substantial genetic diversity (FST = 0.98 and FCT = 0.97). However, the genetic variation is low for the populations of Bella Regia and Bousalem (0.00041), and null between populations of Elfeija, Chebba, Monastir, Sbiba and Sned. Moreover, the genetic distance of the population of Tunis (≥ 0.35) compared to the other populations raises questions about the taxonomic status of this population. Also, in this latter population transitions are the most dominant variations, whereas for the other seven populations there are only two types of transversion substitutions. The results of the current study emphasise the controversial taxonomic status of Porcellionides pruinosus in Tunisia. Consequently, it would appear that the taxonomic status of P. pruinosus from Tunisia needs a re-evaluation.

Genetic differentiation of the purpleback flying squid, Sthenoteuthis oualaniensis , in the South China Sea:population or species divergence

PDF HTML阅读 XML下载 导出引用 引用提醒 南海鸢乌贼的遗传差异: 种群分化还是种间分化 DOI: 作者: 作者单位: 中国水产科学研究院南海水产研究所, 农业农村部外海渔业开发重点实验室, 广东 广州 510300 作者简介: 李敏(1984-),男,博士,副研究员,从事海洋生物多样性和保护研究.E-mail:limin@scsfri.ac.cn 通讯作者: 中图分类号: S931 基金项目: 国家重点研发计划项目（2018YFC1406500）；农业农村部财政专项（NFZX2018）；广东省自然科学基金项目（2014A030310177）；国家重点基础研究发展计划项目（2014CB441500）. Genetic differentiation of the purpleback flying squid, Sthenoteuthis oualaniensis, in the South China Sea:population or species divergence Author: Affiliation: Key Laboratory of Open-Sea Fishery Development, Ministry of Agriculture and Rural Affairs;South China Sea Fisher-ies Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China Fund Project: 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论 摘要:利用线粒体ND2、COI和16S rDNA序列，对2012年9月至2015年9月采自南海的鸢乌贼（）样本进行了种群遗传结构的分析。系统发育关系显示鸢乌贼中型群和微型群各自形成了单系群分支，而分支内没有形成与地理群体或季节群体对应的支系。分子方差分析同样表明，地理群体或季节群体之间不存在明显的遗传分化，而中型群和微型群之间具有极显著的遗传分化（<0.0001）。遗传多样性分析显示，鸢乌贼中型群和微型群均呈现出较高的单倍型多样性（0.9465和0.8545）和较低的核苷酸多样性（0.0051和0.0021），中型群的遗传多样性指数要高于微型群。基于ND2、COI和16S rDNA序列计算的中型群和微型群之间的遗传距离分别达到了14.0%、9.6%和8.8%，远大于类群内的遗传分化（平均<1%）。研究结果表明，南海鸢乌贼存在分布高度重合且有显著遗传分化的两个种群，即中型群和微型群。中型群和微型群之间的遗传差异达到了种间分化的水平，提示体型较大、具有背部发光器的中型群鸢乌贼（典型鸢乌贼）与体型较小、背部发光器缺失的微型群鸢乌贼可能是两个相互独立的种类。 Abstract:In the present study, the population differentiation of the purpleback flying squid () in the South China Sea was examined based on mitochondrial ND2, COI, and 16S rDNA sequences, using samples collected between September 2012 and September 2015. Phylogenetic analysis revealed two significantly divergent monophyletic groups corresponding to the "medium form" and the "dwarf form, " but no geographical lineages corresponding to the sampling localities or seasonal groups were found. The Analysis of Molecular Variance also revealed significant genetic differentiation between the "medium form" and the "dwarf form" (<0.0001), while no genetic variance between either geographic populations or seasonal groups was found. The genetic diversity analysis showed that both the "medium form" and the "dwarf form" of in the South China Sea were characterized by high haplotype diversity (0.9465 and 0.8545) and low nucleotide diversity (0.0051 and 0.0021). The genetic distance between the ‘medium form’ and the "dwarf form" based on ND2, COI, and 16S sequences were 14.0%, 9.6%, and 8.8%, respectively. The genetic distances between the two forms were considerably larger than the genetic distances within each of the two forms (<1%). It was demonstrated that two divergent populations of were sympatrically distributed in the South China Sea, i.e. the "medium form" and the "dwarf form." The genetic differentiation between these two populations was similar to that among separate species within Ommastrephidae, which indicated that the two morphologically distinct forms of the purpleback flying squid, namely the "medium form" (middle sized, with a photophore) and the "dwarf form" (small sized, without a photophore), are two distinct species. 参考文献 相似文献 引证文献

Morphological, Molecular, Biochemical and Nutritional Characterization of Three Major Thais Species from the Sindh Coast of Pakistan

Objective:The present study was conducted to investigate the biomass assessment, morphological and molecular identification, nutritive status and biochemical characterization of three majorThaisspecies (T. bufo,T. hippocastanumandT. rudolphi) from the Sindh Coast, Pakistan.Methods:Samples were collected from Buleji and Paradise Point at the Sindh Coast. Species were identified morphologically as well as genetically by amplifying two mitochondrial 16S rDNA &amp; Cytochrome Oxidase I (COI) and one nuclear (Histone H3) genes. Shell microstructure and chemistry were also studied by scanning electron microscopy and Energy Dispersive X-ray spectrometry (EDX). The body muscle was dissected and used for nutritional composition determination such as estimation of total protein, carbohydrates, lipids, protein fingerprinting by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) and Size-Exclusion - Fast Protein Liquid Chromatography (SEC-FPLC), amino acid and fatty acid analysis.Results:Nutritionally, the total protein was found to be the major content followed by carbohydrate and lipid in the threeThais sp. The presence of medicinally important hemocyanin as abundant hemolymph protein was confirmedviaSDS-PAGE and SEC FPLC. Nine different types of fatty acids and a high concentration of essential amino acids were also determined.Conclusion:Our findings suggest thatThais sp.are nutritionally rich and can be consumed as a valuable marine resource to overcome the malnutrition problem in developing countries.

A Review of the Harvestman Genus Metadentobunus (Opiliones: Sclerosomatidae: Gagrellinae) with A Description of A New Species From Taiwan.

Ssu-Li Chen and Hsi-Te Shih (2018) The harvestman genus Metadentobunus is endemic to Taiwan, and has two taxa, M. formosae formosae Roewer, 1915 and M. formosae garampiensis Suzuki, 1944. The two subspecies have been raised as two species, and this study redescribes the type species M. formosae based on the type specimen and a neotype of M. garampiensis Suzuki, 1944 is assigned. A new species, Metadentobunus brevispinus sp. nov., is also established from central Taiwan based on morphological and molecular evidence. This new species differs from other congeneric species by having one pair of shorter (than other congeners) spines at frontal margin of the ocularium. It can also be distinguished from M. garampiensis by the basal shape of the penile shaft (constricted vs. parallel), location of the seminal receptacle in ovipositor (5th-6th segment vs. 6th-7th segment) and number of ampullas in the terminal of the seminal receptacle (2 vs. 3). Molecular evidence from mitochondrial 16S rDNA and cytochrome oxidase subunit I (COI) further supports the identity of this new species. An identification key for this genus is included as well.

GoFish: A versatile nested PCR strategy for environmental DNA assays for marine vertebrates.

Here we describe GoFish, a strategy for single-species environmental DNA (eDNA) presence/absence assays using nested PCR. The assays amplify a mitochondrial 12S rDNA segment with vertebrate metabarcoding primers, followed by nested PCR with M13-tailed, species-specific primers. Sanger sequencing confirms positives detected by gel electrophoresis. We first obtained 12S sequences from 77 fish specimens for 36 northwestern Atlantic taxa not well documented in GenBank. Using these and existing 12S records, we designed GoFish assays for 11 bony fish species common in the lower Hudson River estuary and tested seasonal abundance and habitat preference at two sites. Additional assays detected nine cartilaginous fish species and a marine mammal, bottlenose dolphin, in southern New York Bight. GoFish sensitivity was equivalent to Illumina MiSeq metabarcoding. Unlike quantitative PCR (qPCR), GoFish does not require tissues of target and related species for assay development and a basic thermal cycler is sufficient. Unlike Illumina metabarcoding, indexing and batching samples are unnecessary and advanced bioinformatics expertise is not needed. From water collection to Sanger sequencing results, the assay can be carried out in three days. The main limitations to this approach, which employs metabarcoding primers, are the same as for metabarcoding, namely, inability to distinguish species with shared target sequences and inconsistent amplification of rarer eDNA. In addition, the performance of the 20 assays reported here as compared to other single-species eDNA assays is not known. This approach will be a useful addition to current eDNA methods when analyzing presence/absence of known species, when turnaround time is important, and in educational settings.

Versuriga anadyomene, a newly recorded scyphozoan jellyfish (Scyphozoa: Rhizostomae) in Chinese waters

Scyphozoans are currently causing great concern worldwide due to the effects of their blooms on the marine environment. During a field survey in Beibu Gulf, South China Sea, on 27 May 2015, we found specimens of a rhizostome jellyfish that had not previously been recorded in Chinese seas. The morphological characters and DNA sequences of several specimens were examined in the laboratory. These specimens had the following morphological characters: hemispherical umbrella with reticular grooves; eight large semicircular velar lappets alternating with narrow lappets in each of the 8 octants; laterally flattened mouth-arms with numerous flat, membranous branches bearing many clubs and intermediate filaments; non-anastomosed broad perradial canals and anastomosed narrow interradial canals. The morphological characters of these rhizostome jellyfish were consistent with descriptions of Versuriga anadyomene. Furthermore, phylogenetic analysis of mitochondrial 16S rDNA and nuclear 18S rDNA gene fragments confirmed that these specimens were in the same cluster as V. anadyomene collected in Palau. Therefore, this rhizostome jellyfish was identified as V. anadyomene. This is the first record of the genus Versuriga and the species V. anadyomene in Chinese seas. The arrival of this species may be attributed to physical transport from the coastal waters of South East Asia.

Phylogeny of Tricorythodes Ulmer (Leptohyphidae: Ephemeroptera) based on molecular and morphological evidence

The first phylogeny of Tricorythodes Ulmer (Ephemeroptera: Leptohyphidae) based on molecular and morphological evidence is presented. A parsimony analysis was conducted with 56 morphological (24 continuous and 32 discrete) characters of 48 species, 39 belonging to Tricorythodes sensu lato and nine additional groups. Bayesian and maximum likelihood analyses of molecular and combined molecular and morphological (32 discrete) data included a total of 22 taxa for which it was possible to extract DNA. DNA sequences used correspond to fragments of the nuclear 18S rDNA and the mitochondrial COI and 16S rDNA (total of 1804 bp). Tricorythodes sensu lato monophyly was tested, as well as validity of some recently proposed related genera. Results of this work fully support the synonymy of Epiphrades and Tricorythodes, as suggested by previous authors. Part of the analyses also support the synonymy of Asioplax and Homoleptohyphes with Tricorythodes, but they change position radically when morphology alone (parsimony) and the combined data (Bayesian and maximum likelihood) are analyzed. Furthermore, Tricoryhyphes was recovered as a polyphyletic group and Cabecar and Loricyphes appeared nested well within Tricorythodes sensu lato. Monophyletic groups of species inside this large Panamerican genus were found. A phylogenetic framework for the group is much needed to better understand the evolution of disparate nymphal body form, ecology and biogeography inside this genus.

A Redescription of the Stygobitic Amphipod Crangonyx grandimanus (Amphipoda: Crangonyctidae) Including Phylogenetically Significant Sequence Data for Mitochondrial and Nuclear Genes

Crangonyx grandimanusBousfield, 1963, a relatively large stygobitic amphipod endemic to the Floridan aquifer, is redescribed from its holotype specimen and topotype material. In addition to the morphological redescription, genetic sequence data for sections of the nuclear 18S rDNA, 28S rDNA, and mitochondrial 16S rDNA are provided from the examined topotypes. The erratum for this article can be found in Bulletin of the Peabody Museum of Natural History, Vol. 60 No. 1.

Relationships of the Sphaeromatidae genera (Peracarida: Isopoda) inferred from 18S rDNA and 16S rDNA genes

The Sphaeromatidae has 100 genera and close to 700 species with a worldwide distribution. Most are abundant primarily in shallow (&lt; 200 m) marine communities, but extend to 1.400 m, and are occasionally present in permanent freshwater habitats. They playan important role as prey for epibenthic fishes and are commensals and scavengers. Sphaeromatids’ impressive exploitation of diverse habitats, in combination with diversity in female life history strategies and elaborate male combat structures, has resulted in extraordinary levels of homoplasy. We sequenced specimens from 39 genera for nuclear 18S rDNA and mitochondrial 16S rDNA genes, comprehensively reviewed the effects of alignments on tree topology, and performed Garli and MrBayes analyses. These data consistently retrieved clades (genus groups), Sphaeroma, Exosphaeroma, Cymodoce, Ischyromene, Cerceis, and Dynamenella and the monogeneric clade of Gnorimosphaeroma. We define the major clades using morphological characters, attribute sampled taxa to consistently and strongly supported ones and suggest placement of unsampled genera based on their morphological characteristics. Within each clade, we also highlight unresolved and poorly sampled genera. We point out taxonomic problems in hopes of encouraging further phylogenetic exploration. Although we identify clades containing consistent generic groups and are confident that some groups will prove stable and reliable, we feel our sampling is insufficient to propose nomenclatural changes at this time.

A Combined Molecular and Morphological Approach to Explore the Higher Phylogeny of Entimine Weevils (Coleoptera: Curculionidae), with Special Reference to South American Taxa

The Entiminae are broad-nosed weevils constituting the most diverse subfamily of Curculionidae, with over 50 tribes. We performed Bayesian and Maximum Parsimony combined phylogenetic analyses with the main objective of testing higher-level relationships and the naturalness of the major Neotropical and Southern South American (Patagonia and Andes) tribes, including some members from other regions. We compiled a data matrix of 67 terminal units with 63 Entiminae species, as well as four outgroup taxa from Cyclominae, by 3522 molecular (from nuclear 18S rDNA and 28S rDNA, and mitochondrial 16S rDNA and COI gene sequences) and 70 morphological characters. The resulting trees recover a clade Entiminae with a monophyletic Cylydrorhinini and Premnotrypes branching off early. The tree resulting from parsimony analysis shows a clade of Leptopiini from the Australian region and another clade including taxa mainly distributed in the Palaearctic and Neotropical regions, but in the Bayesian tree the South American and Australian Leptopiini are grouped together. The mainly Palaearctic Entiminae (e.g., Brachyderini, Laparocerini, Otiorhynchini, Peritelini, Polydrusini, Phyllobiini and Sciaphylini) form a subclade separated from Southern Hemisphere taxa. Among the latter, the well-supported Naupactini are the sister group of the South American Tanymecini, excluding Platyaspistes, herein transferred to Leptopiini (new placement). Another well-justified clade is Eustylini–Geonemini, which also includes the enigmatic Galapagonotus, and the genus Artipus, thus corroborating its recent exclusion from Naupactini.

Development of a DNA metabarcoding method for the identification of fifteen mammalian and six poultry species in food

Meat products are prone to adulteration by the replacement of meat from more expensive animal species with meat from cheaper sources. We present a DNA metabarcoding method allowing the identification and differentiation of 15 mammalian and six poultry species in foodstuffs. The method, developed on the MiSeq® platform, targets a mitochondrial 16S rDNA region recently found to be suitable for the differentiation of 300 mammalian species. We designed a novel primer pair for poultry and applied it in combination with the primer pair for mammalian species in a duplex assay. The applicability of the method was investigated by analysing DNA extracts from muscle, DNA extract mixtures and extracts from model sausages. Our results indicated that the species of interest can be identified, differentiated and detected down to a proportion of 0.1%. Since 96 samples can be sequenced in one run, the method has high potential for application in routine analysis.

Mitochondrial DNA analyses of Donax trunculus (Mollusca: Bivalvia) population structure in the Iberian Peninsula, a bivalve with high commercial importance

Abstract The wedge clam, Donax trunculus (Linnaeus, 1758), is an ecologically and economically important species found in the warm waters of the Atlantic and Mediterranean areas. Donax trunculus has been suffering from a severe population decline in some wild stocks of the Iberian Peninsula, and genetic monitoring of wild populations can provide valuable information to support conservation measures and management policies. In this study, mitochondrial 16S rDNA (16S) and cytochrome b (Cytb) gene variation was examined at 18 localities across the Iberian Peninsula to test whether the patterns of genetic diversity, population structure, and demographic inferences previously deduced by using microsatellites are supported by these molecular markers. Samples (N = 557) were collected from 18 localities of the Iberian Peninsula and attention was paid to the north‐western Atlantic area, where wild stocks have diminished greatly. Analysis of molecular variance, fixation index (FST), and demographic inferences were estimated based on mitochondrial DNA. Since D. trunculus shows doubly uniparental inheritance, both types of mitochondrial DNA (F and M) were analysed. A high level of genetic diversity was detected using the haplotype diversity dataset, showing different geographic patterns. FST values for the M type revealed significant differentiation between populations on both sides of the Almeria–Oran front, and between the locality from the Alboran Sea (FU) and localities from the Atlantic Ocean. However, the F type did not detect the differentiation of FU. This makes the M genome sequences more suitable for investigating genetic structure of D. trunculus populations. Management and recovering strategies of this important resource should consider the genetic structure observed. Several strategies for conservation and management of this commercial and ecologically important bivalve are proposed.

Detection of Rickettsia spp. in ticks parasitizing toads (Rhinella marina) in the northern Brazilian Amazon.

This study evaluated rickettsial infection in ticks collected on toads from the northern Brazilian Amazon (Amapá state), where to our knowledge there are neither records of ticks from amphibians nor rickettsial infections in ticks. During 2016-2017, a total of 22 free-living toads were captured and identified as Rhinella marina. Overall, 12 (54.5%) toads were parasitized by a total of 97 ticks (6 males, 39 females, 31 nymphs, 21 larvae), giving a mean intensity of 8.1 ticks per infested toad. Two tick species were morphologically identified: Amblyomma rotundatum Koch (31 females, 14 nymphs), and Amblyomma dissimile Koch (6 males, 8 females, 17 nymphs). The 21 larvae were morphologically denoted as Amblyomma sp. Five toads were co-infested by A. rotundatum and A. dissimile. Morphological identifications were confirmed by nucleotide sequencing of fragments of the mitochondrial genes 16S rDNA, 12S rDNA and/or COX1. A total of 54 ticks were analyzed for the presence of rickettsial DNA. Eleven (9 females and 2 nymphs) out of 14 A. rotundatum ticks contained Rickettsia bellii. None of the 25 specimens of A. dissimile (6 males, 6 females, 13 nymphs) contained amplifiable rickettsial DNA. From 15 Amblyomma sp. larvae, a pool of 10 individuals contained Rickettsia sp. strain Colombianensi. Sequencing of the 16S rDNA amplicon derived from the positive pool yielded a sequence of A. dissimile. We detected Rickettsia sp. strain Colombianensi for the first time in Brazil. Prior records of this agent were restricted to Colombia and Honduras. In addition, we report the presence of A. rotundatum for the first time in the state of Amapá, where the only other record of A. dissimile was registered over 20years ago.

&lt;p class="Body"&gt;Amblyomma yucumense Krawczak, Martins &amp;amp; Labruna, 2015 (Acari: Ixodidae) in Argentina

Amblyomma yucumense was recently described from specimens collected in the Turvo State Park, southern Brazil. In search of this tick species, free-living ticks were collected at the Moconá Provincial Park and the Yabotí Biosphere Reserve in Misiones province, Argentina. A total of 594 ticks were collected, of which 448 were Amblyomma brasiliense, 90 Amblyomma incisum, one Amblyomma ovale, 48 Haemaphysalis juxtakochi and seven A. yucumense. The latter was confirmed by the presence of certain morphological characters and the analysis of sequences of a 410-bp fragment of the mitochondrial 16S rDNA performed to representative specimens. The sequences showed 100% identity with that of A. yucumense from Turvo State Park in Brazil. Amblyomma yucumense, Amblyomma dubitatum and Amblyomma coelebs phylogenetically group together and are morphologically similar, though they seem to have distinct ecological preferences. Amblyomma coelebs has only been found in northern Misiones province, while the most numerous records of A. dubitatum come from the central area of the province. Until now, the distribution of A. yucumense in Argentina is restricted to Moconá and Yabotí. The findings in this study expand A. brasiliense, A. incisum, A. ovale and H. juxtakochi distribution ranges to the south, and we report for the first time the presence of A. yucumense in Argentina.

A discreet invasion: distribution, origins, and expansion of the European slug complexArion subfuscuss.l. in Quebec

Accurate identification of species and their geographic origins is crucial to assess the impacts of exotic species. This is particularly relevant when multiple introductions are suspected. Species complexes are therefore problematic, as different species can be treated as a single taxon. The highly diversified European complex Arion subfuscus (Draparnaud, 1805) s.l. is abundant throughout northeastern North America. However, surveys performed in the United States have reported a low mitochondrial diversity associated with a single species. To assess whether regions characterized by different human activities harbor different diversities, the taxonomic identities, distributions, and origins of the species of this complex in Quebec (Canada) were determined. The mitochondrial 16S rDNA gene was analyzed in 526 specimens from 69 locations. Both Arion fuscus (O.F. Müller, 1774) and Arion subfuscus (Draparnaud, 1805) s. str. (lineages S1 and S2) were detected. Arion fuscus was the most abundant species, contrasting with previous studies conducted in North America. An explosive range expansion occurred during the last 50 years. Moreover, the presence of different haplotypes confirmed multiple introductions of these species. These results emphasize the importance of not considering an exotic species complex as a single taxon, especially in regions with different human activities.