Partial Cytochrome Oxidase Research Articles

Prevalence and distribution of Taenia solium cysticercosis in naturally infected pigs in Punjab, India.

BackgroundTaenia solium (T. solium) cysticercosis remains a neglected zoonotic disease in India. The current study was planned to estimate the prevalence of T. solium porcine cysticercosis in the Punjab state of India, to compare this prevalence with the disease prevalence in pigs reared outside Punjab and to assess the distribution of the parasite in pig carcasses.MethodsTwo slaughter shops were selected in each of the 22 districts of Punjab. Pigs slaughtered on the day/s of inspection were post-mortem inspected to identify the presence of T. solium cysts. Estimated true prevalence was estimated by taking into account the diagnostic sensitivity (38%) and specificity (100%) of post-mortem inspection using the Rogan-Gladen estimator. Positive carcasses were purchased and brought to the laboratory to assess the tissue distribution of T. solium cysts and to conduct PCR targeting large subunit rRNA gene, internal transcribed spacer 1 gene, ITS1 gene and Cytochrome oxidase I gene. The selected PCR products were submitted for sequencing and phylogenetic analyses were performed.FindingsWe contacted 71 shop owners to achieve a sample of 44 shops for the study. We inspected 642 pigs reared in Punjab and 450 imported from other states at these slaughter shops. In addition, we sampled 40 pigs from an abattoir located in the state capital. Of the 642 pigs reared in Punjab, 9 had T. solium cysts with an apparent prevalence of 1·40% (95% CI: 0·74%, 2·64%) and the estimated true prevalence of 3.69% (95% CI: 1·95%, 6·95%). Pigs imported from outside the state had a significantly higher prevalence (odds ratio: 2·58; 95% CI: 1·12, 5·98; p-value: 0·026) as 15 of the 450 imported pigs were positive (apparent prevalence: 3.33%; 95% CI: 2.03%, 5.43%; estimated true prevalence: 8.77%; 95% CI: 5.34%, 14.28%). None of samples was positive from the pigs sampled at the abattoir in the state capital. The PCR confirmed T. solium cysts from all the 24 positive samples. We counted a median of 897 (range 526–1964) cysts per infected pig from the 19 infected pig carcasses inspected. The phylogenetic tree based on the alignment of partial cytochrome oxidase 1 sequences indicated all positive samples to be clustered with the T. solium Asian genotype. The analysis did not indicate the presence of T. asiatica in the slaughter pigs.ConclusionsDespite the underestimation of the prevalence due to missing mildly-infected carcasses, low participation and lack of representative sampling, the presence of heavily infected carcasses containing viable cysts, particularly those imported from outside the state, indicates that T. solium cysticercosis is an important food safety concern for pork consumers in Punjab, India. Measures should be taken to reduce the disease prevalence in pigs to reduce the disease burden in the public.

Determination of Genetic Diversity in Chilo partellus, Busseola fusca, and Spodoptera frugiperda Infesting Sugarcane in Southern Malawi Using DNA Barcodes.

Sugarcane is one of the most valuable crops in the world. Native and exotic Lepidopteran stemborers significantly limit sugarcane production. However, the identity and genetic diversity of stemborers infesting sugarcane in Malawi is unknown. The main objectives for this study were to identify and determine genetic diversity in stemborers infesting sugarcane in Malawi. We conducted field surveys between June 2016 and March 2017 in the Lower Shire Valley district of Chikwawa and Nsanje, southern Malawi. Molecular identification was based amplification the partial cytochrome oxidase subunit I (COI) gene region. Phylogenetic trees for sequences were generated and published GenBank accessions for each species were constructed. We found that Malawi Busseola fusca (Lepidoptera: Noctuidae) specimens belonged to clade II, Spodoptera frugiperda sp. 1 (Lepidoptera: Noctuidae) and Chilo partellus (Lepidoptera: Crambidae) were infesting sugarcane. Interspecific divergence ranged from 8.7% to 15.3%. Intraspecific divergence was highest for B. fusca, 3.6%. There were eight haplotypes for B. fusca, three for S. frugiperda and three for C. partellus. The importance of accurate species identification and genetic diversity on stemborer management is presented.

First Report of the Yellow Nutsedge Cyst Nematode, Heterodera cyperi, in Georgia, U.S.A.

Soil samples collected during a survey for plant-parasitic nematodes in Tift County GA in summer 2017 were submitted for routine diagnosis of nematodes to the Extension Nematology Lab at the Department of Plant Pathology, University of Georgia, Athens, Georgia. Cyst nematodes recovered by centrifugal flotation technique were discovered in the samples from two research sites in a field with a history of tobacco and vegetable production. Cyst nematodes from tobacco (10 cysts/100 cm 3 of soil) and vegetable (2 cysts/100 cm 3 of soil) sites had similar morphological features. Morphology and morphometric measurements of the cysts and J2 ( Fig. 1A-C ) were in agreement with those of Heterodera cyperi ( Golden et al., 1962 ; Romero and López-Llorca, 1996 ). Measurements of J2 ( n = 12) included the length (range = 443-494 μm, mean = 467.4 μm) and width (18.3-24.4 μm, 20.6 μm) of body, stylet (19.1-20.8 μm, 20.3 μm), tail (61.6.0-66.4 μm, 64.2 μm), body width at anus (11.9-14.1 μm, 12.8 μm), and hyaline tail terminus (22.7-29.2 μm, 26.3 μm). The lateral field of J2 had three lines. Cysts ( n = 10; Fig. 1C ) were lemon-shaped, light to dark brown in color with protruding neck and vulval cone. The cysts had ambifenestrated vulval cone and no bullae was present. Morphometrics included body length excluding neck (370.5-714.4 μm, 555.7 μm); body width (165.6-411.1 μm, 310.9 μm); neck length (36.5-66.3 μm, 49.8 μm); fenestra length (26.3-42.5 μm, 35.8 μm), and fenestra width (19.1-31.5 μm, 23.8 μm). DNA was extracted from single cysts ( n = 3) and internal transcribed spacer (ITS) of rRNA and partial cytochrome oxidase I ( COI ) genes were amplified with primers TW81/AB28 and Het-coxiF/Het-coxiR, respectively ( Subbotin et al., 2001 ; Subbotin, 2015 ) and sequenced. The resulting sequences were deposited into the GenBank database (Accession no. MG825344 and MG857126) and also subjected to BLAST searches in the database. ITS sequence of H. cyperi showed 100% similarity (100% coverage) with that of a H. cyperi population from Spain (AF274388). COI sequence of H. cyperi showed 89% similarity (98% coverage) with that of H. guangdongensis (MF425735), and 88% similarity (83% coverage) with that of H. elachista (KC618473). The pathogenicity of H. cyperi was examined under greenhouse conditions using tobacco cv. K340, tomato cv. Tribute, cucumber cv. Thunder, and yellow nutsedge ( Cyperus esculentus L.). 3-wk-old seedlings of the test plants were transferred into Deepot D25L cell containers (5-cm-diam. × 25.4-cm deep) filled with sterilized sand: sand: soil mixture (1:2) and then inoculated with 1,000 eggs and J2 of H. cyperi . The plants were grown for 90 d in a greenhouse before examination of roots and extraction of cysts from the soil. Results showed that the nematode failed to reproduce on tobacco, tomato, and cucumber whereas white females and mature cysts of H. cyperi were observed on yellow nutsedge roots ( Fig. 1E ). The results confirmed that yellow nutsedeg was a host for the nematode, and tobacco, tomato, or cucumber were non-hosts. In the United States, H. cyperi was reported from Florida, North Carolina, and Arkansas ( Subbotin et al., 2010 ) infecting Cyperus spp. Yellow nutsedge is considered a serious weed problem in many cropping systems including peanut, cotton, tobacco, and vegetable crops in the Southern United States. To our knowledge, this is the first report of H. cyperi infecting yellow nutsedge in Georgia. Figure 1Photomicrographs of Heterodera cyperi from yellow nutsedge in Georgia. Whole body (A), the anterior region (B), and the posterior region (C) of J2. Cysts (D) recovered from the soil and the vulval cone of cyst with the ambifenestrate fenestra (E). A mature cyst (F) on the surface of yellow nutsedge root infected with the nematode.

Identification and phylogenetic analysis of fruit borer species of litchi using DNA barcode sequences

Fruit borers are the major insect pests of litchi in humid orchard conditions at the time of fruit ripening in Indian subcontinent. The major constraint faced in formulating any management strategies against the borer is the difficulty in identification of the correct species. The litchi fruit borer has low intraspecific variation among the existing borer populations. Molecular approaches have been used to identify various species. In the present study, partial cytochrome oxidase I (COI) sequences were used to understand the phylogenetic relationship among borer complex, and assess their usefulness to identify and classify unknown borer species collected from litchi orchards. All together, 150 specimens of litchi fruit borer from Bihar and Jharkhand were examined and 2 morphologically similar moths of each genus were used for further analysis. Sequence analysis revealed that the intraspecific and interspecific variations ranged from zero to 10.0% and 4.4 to 20.3%, respectively among fruit borer complex. The phylogenetic analysis showed that the borers specimen used in this study clustered in to distinct species-groups designated as Conopomorpha sinensis Bradley, C. litchiella Bradley, Cryptophlebia ombrodelta Lower and Gatesclarkeana spp. Higher intraspecific genetic variation was observed in Conopomorpha species complex as compared to Cryptophlebia species complex. Present study has clearly demonstrated that DNA barcoding is an efficient and accurate method for identification of coexists of borer complex infestation in litchi fruits. Hence, this approach can play important role in formulating viable pest management strategies.

A new orthopteran-parasitizing horsehair worm, Acutogordius taiwanensis sp. n., with a redescription of Chordodes formosanus and novel host records from Taiwan (Nematomorpha, Gordiida).

A description of a new species of horsehair worm, Acutogordius taiwanensis sp. n., a redescription of Chordodes formosanus, and novel host records for the latter are provided. Acutogordius taiwanensis sp. n. is morphologically similar to A. protectus with moderately flat areoles on its tail tips, but is distinguishable by small mid-body ornamentations. Despite the distinct differences in the post-cloacal crescents between 14 male samples, their conspecific status, along with that of nine female samples, was upheld by a phylogenetic comparison of partial cytochrome oxidase subunit I (COI) sequences. Chordodes formosanus is another common horsehair worm species in Taiwan, which was previously believed to specifically parasitize Hierodula mantids. However, in this study, five C. formosanus were observed emerging from an Acromantis mantid, and two long-horned grasshopper hosts (Leptoteratura sp. and Holochlora japonica). These five worms showed high degrees of similarity in COI sequences and morphology, but one of these individuals bore abnormal crowned areoles, which has never been observed in C. formosanus, and may be attributed to the incomplete development of this particular individual.

Prospects on the Application of DNA Barcoding on Soil-Transmitted Helminths in Children

AbstractSince more than a decade, DNA barcoding has been widely used to examine biological samples and differentiate species, as well as employed in ecological and conservational studies. There is a growing interest of DNA barcoding, particularly in medical parasitology, but its potential utility in soil-transmitted helminths (STHs) remains unclear. Therefore, in this article, we review the studies using DNA barcoding and its applications in medical parasitology with special focus on STHs such as Ascaris lumbricoides, Ancylostoma duodenale, Necator americanus, and Trichuris trichiura. DNA barcoding is reliable for identifying STH species as well as its cryptic species. In addition, epidemiological data and the impacts of STH infections on children are discussed. This article further discusses the paucity of STH DNA barcodes (partial cytochrome oxidase subunit 1 [COI] mitochondrial DNA [mtDNA] sequences) in two gene banks; National Center for Biotechnology Information (NCBI) and Barcode of Life Data (BOLD) Systems. It also highlights the future prospects of DNA barcoding including primer designing and reference library on STHs.

Description of the Asian chili pod gall midge, Asphondylia capsicicola sp. n., with comparative notes on Asphondylia gennadii (Diptera: Cecidomyiidae) that induces the same sort of pod gall on the same host plant species in the Mediterranean region

A new species of the genus Asphondylia (Diptera: Cecidomyiidae) that infests pods of chili, Capsicum annuum L. and Capsicum frutescens L. (Solanaceae), is described as Asphondylia capsicicola sp. n. based on specimens collected from Indonesia and Vietnam. The new species is similar to Asphondylia gennadii (Marchal) (=Asphondylia capsici) that induces chili pod galls in the Mediterranean region, but is distinguishable from it by the morphological features of pupa such as the nonlinear arrangement of the lower frontal horns, and the narrower longitudinal band of transverse wrinkles on the tergite of the mesothorax. Differences between the two species in the DNA sequencing data were 69 bp (16%) to 77 bp (19%) among 413 bp of the partial cytochrome oxidase subunit I region examined, supporting the morphological identification. This is one of the examples in which two congeners induce the same sort of gall on the same host plant organ and species, which means that the two species are not distinguishable based solely on gall shape and host plant information, unlike many other gall midges.

Preliminary Taxonomic Survey and Molecular Documentation of Jellyfish Species (Cnidaria: Scyphozoa and Cubozoa) in Malaysia.

Mohammed Rizman-Idid, Abu Bakar Farrah-Azwa, and Ving Ching Chong (2016) Scientific enquiries into jellyfish blooms and associated problems are often deterred by the lack of taxonomical and ecological studies worldwide. Taxonomic difficulty is attributed to the high degree of morphological variations among and within species. To date, only two scyphozoan jellyfish species have been documented from field surveys in Malaysian waters, whereas another four Malaysian scyphozoan and two cubozoan jellyfish species have been mentioned in toxicological studies. None of these species have; however, been verified. This study thus aimed to document and resolves the uncertainty of earlier identified species in the region using morphology and molecular DNA sequencing. Jellyfish specimens were collected from Malaysian waters in the Straits of Malacca, South-China Sea and the Sulu-Sulawesi Sea over two years (June 2008 to October 2010), and their DNA sequences were compared with those from the Atlantic and Pacific regions. Ten scyphozoan and two cubozoan species were recorded in Malaysian waters (South China Sea and Straits of Malacca). These jellyfish included eight species from the order Rhizostomeae (Rhizostomatidae, Lobonematidae, Mastigiidae, Catostylidae and Cepheidae), two species from Semaestomeae (Pelagiidae and Cyaneidae) and two species from class Cubozoa; one from order Carybdeida (family Carukiidae) and another from order Chirodropida (family Chiropsalmidae). Molecular identification of species using phylogenetic approaches was based on DNA sequences of partial cytochrome oxidase I (COI), 16S and internal transcribed spacer (ITS1) regions. The COI phylogenetic tree of Cubozoa and Scyphozoa species from the Atlantic and Pacific regions showed distinct clustering of six Malaysian jellyfish species. However, most of the deeper divergences and relationships between the families were unresolved, which were also observed in the 16S and ITS1 phylogenetic trees. The Malaysian edible species Lobonemoides robustus, Rhopilema hispidum and Rhopilema esculentum were grouped within Rhizostomeae, whereas other scyphozoans showed phylogenetic affinities to Semaestomeae and Kolpophorae. Chrysaora and Cyanea appeared non-monophyletic; however their paraphyly was not confirmed. This study has provided the much needed baseline information on the taxonomy of Malaysian jellyfish species which have been substantiated by partial COI, 16S and ITS1 sequences. A total of 12 putative species of jellyfish were identified, which encompassed 12 genera.

Soybean Stem Fly, Melanagromyza sojae (Diptera: Agromyzidae), in the New World: detection of high genetic diversity from soybean fields in Brazil.

Soybean Stem Fly (SSF), Melanagromyza sojae (Zehntner), belongs to the family Agromyzidae and is highly polyphagous, attacking many plant species of the family Fabaceae, including soybean and other beans. SSF is regarded as one of the most important pests in soybean fields of Asia (e.g., China, India), North East Africa (e.g., Egypt), parts of Russia, and South East Asia. Despite reports of Agromyzidae flies infesting soybean fields in Rio Grande do Sul State (Brazil) in 1983 and 2009 and periodic interceptions of SSF since the 1940s by the USA quarantine authorities, SSF has not been officially reported to have successfully established in the North and South Americas. In South America, M. sojae was recently confirmed using morphology and its complete mitochondrial DNA (mtDNA) was characterized. In the present study, we surveyed the genetic diversity of M. sojae, collected directly from soybean host plants, using partial mtDNA cytochrome oxidase I (COI) gene, and provide evidence of multiple (>10) maternal lineages in SSF populations in South America, potentially representing multiple incursion events. However, a single incursion involving multiple-female founders could not be ruled out. We identified a haplotype that was common in the fields of two Brazilian states and the individuals collected from Australia in 2013. The implications of SSF incursions in southern Brazil are discussed in relation to the current soybean agricultural practices, highlighting an urgent need for better understanding of SSF population movements in the New World, which is necessary for developing effective management options for this significant soybean pest.

Phylogeny and Genetic Divergence of Indian Himalayan Population of Anomala dimidiata (Coleoptera: Scarabaeidae) Inferred from Mitochondrial DNA Sequences

Anomala dimidiata, a key white grub species associated with substantial crop losses in Indian Himalayan region, belong to one of the largest genera under scarab tribe, Anomalini. This paper presents the molecular variation and phylogenetic relationship of this species from the region using partial cytochrome oxidase I (COI) and cytochrome b (cyt b) sequences of mitochondrial genome in both nucleotides and polypeptides. The COI sequences were compared with available five sequences of A. dimidiata and 12 sequences of species under genus Anomala to study the divergence within and between species, respectively. However, cyt b sequence, as we were presenting for the first time, was compared with cyt b sequences of Drosophila yakuba whose entire mitochondrial genome was known and Dasylepida ishigakiensis, a scarabid white grub species, whose partial cyt b sequence is available. Both the genes were A + T biased and showed differential codon usage between species and geographical isolates. This is evidenced by discrepancies between nucleotide and deduced polypeptide sequences. The phylogenetic relationship of COI sequences also revealed that A. dimidata was evolutionarily associated with A. xanthoptera. The evolutionary distances of three cyt b sequences showed that differences were larger than expected based on evolutionary divergence. The study illustrates a complex genetic variation coupled with highly structured evolutionary divergences between and within species of Anomala.

A new species of Mesochaetopterus (Annelida, Chaetopteridae) from Hong Kong, with comments on the phylogeny of the family

We described a new species, Mesochaetopterus tingkokensis, based on 14 specimens collected from an intertidal area in Hong Kong. This species is large (body length of complete specimens 17.9-24.1 cm), with 9, 2 and 36-41 chaetigers in region A, B and C, respectively. It belongs to a small group of Mesochaetopterus species with an expanded wing-shaped notopodia in chaetiger B2. The new species can be distinguished from other Mesochaetopterus species in this group by having a pair of palps with two longitudinal stripes formed by suture-like discontinuous orange bands, more teeth in the uncini of region B and C neuropodia, and presence of a bundle of simple chaetae in region C notopodia. Comparison with other chaetopterids based on partial cytochrome oxidase I (COI), 18S and 28S rRNA gene sequences confirmed the placement of M. tingkokensis n. sp.within Mesochaetopterus and its distinction from other members of this genus with the available DNA sequences. The phylogenetic tree base on COI showed that Mesochaetopterus and Chaetopterus are paraphyletic, but that based on concatenated data, 18S and 28S showed they are monophyletic with low supporting values.

Taiwanassiminea phantasma n. sp.: a terrestrial assimineid (Caenogastropoda: Truncatelloidea) from Middle Osborn Island, Kimberley, Western Australia

A new terrestrial assimineid gastropod, Taiwanassiminea phantasma n. sp. (Truncatelloidea: Assimineidae), is described. Taiwanassiminea phantasma is characterised by its translucent, conical shell with deeply impressed sutures, spiral groove surrounding a wide umbilicus, relative lack of pigmentation and long distal penial papilla. The species differs from most assimineids in having a subcircular aperture and an operculum with a central nucleus. Taiwanassiminea phantasma has anatomical similarities to other members of Taiwanassiminea, and this relationship is supported by a partial cytochrome oxidase I mitochondrial analysis. Taiwanassiminea phantasma lives in leaf litter in vine thickets on Middle Osborn Island in northern Kimberley, Western Australia, where it is possibly endemic. No terrestrial assimineid is otherwise known from mainland Australia or its adjacent islands, including Tasmania.http://zoobank.org/urn:lsid:zoobank.org:pub:A14ACAAE-4BAD-402D-9F9D-3BC9D928411A

Angiostrongylus chabaudi Biocca, 1957: a new parasite for domestic cats?

BackgroundNatural infection with a species of Angiostrongylus has been reported only once in wildcats from central Italy by Biocca in 1957. The causative species of this infection was identified as Angiostrongylus chabaudi. Following this report, this parasite had never been found in either wild or domestic cats.FindingsThe lungs and the pulmonary arteries of an adult female cat (Felis silvestris catus), road-killed in Sardinia, Italy, were macroscopically examined and dissected under a light microscope for the presence of parasites. A slender nematode was detected and its morphometrical features were consistent with those of A. chabaudi. Morphological data were supplemented by sequencing of the partial cytochrome oxidase c subunit 1 (cox1) gene, as well as the internal transcribed spacer 2 (ITS2) of the rDNA. Nucleotide sequences displayed 99% homology with the ITS2 sequence [GenBank KM216825.1] of a specimen of Angiostrongylus sp. recovered recently from the pulmonary artery of a wildcat in Germany and 91% with cox1 sequence [GenBank GU138118.1] of Angiostrongylus vasorum.ConclusionThe results of the present study indicate, for the first time, that A. chabaudi may also infect domestic cats, and thus should be considered in the diagnosis of metastrongyloid species infecting their cardio-pulmonary system.

First Report of Phytophthora hydropathica Causing Wilting and Shoot Dieback on Viburnum in Italy

The genus Viburnum comprises over 150 species of shrubs and small trees such as Laurustinus (Viburnum tinus L.), which is one of the most widely used ornamental plants in private and public gardens. Furthermore, it commonly forms stands of natural woodland in the Mediterranean area. In autumn 2012, a survey was conducted to determine the presence of Phytophthora ramorum on Viburnum in commercial nurseries in the Latium region where wilting, dieback, and death of twigs were observed on 30% of the Laurustinus plants. A Phytophthora species was consistently recovered from soil rich in feeder roots from potted Laurustinus plants showing symptoms. Soil samples were baited with rhododendron leaves. Small pieces of leaf tissue cut from the margin of lesions were plated on P5ARPH selective medium (4). Pure cultures, obtained by single-hypha transfers on potato dextrose agar (PDA), were petaloid. Sporangia formation was induced on pepper seeds (3). Sporangia were almost spherical, ovoid or obpyriform, non-papillate and non-caducous, measuring 36.6 to 71.4 × 33.4 to 48.3 μm (average 53.3 × 37.4 μm) with a length/width ratio of 1.4. Chlamydospores were terminal and 25.2 to 37.9 μm in diameter. Isolates were considered heterothallic because they did not produce gametangia in culture or on the host. All isolates examined had 30 to 35°C as optimum temperatures. Based on these morphological characteristics, the isolates were identified as Phytophthora hydropathica (2). Morphological identification was confirmed by internal transcribed spacer (ITS), and mitochondrial partial cytochrome oxidase subunit 2 (CoxII) with BLAST analysis in the NCBI database revealing 99% identity with ITS and 100% identity with CoxII. The sequences of the three isolates AB234, AB235, and AB236 were deposited in European Nucleotide Archive (ENA) with the accession nos. HG934148, HG934149, and HG934150 for ITS and HG934151, HG934152, and HG934153 for CoxII, respectively. Pathogenicity tests were conducted in the greenhouse on a total of six 1-year-old shoots cut from V. tinus plants with two inoculation points each. Mycelial plugs cut from the margins of actively growing 8-day-old cultures on PDA were inserted through the epidermis into the phloem. Controls were treated as described above except that sterile PDA plugs replaced the inoculum. Shoots were incubated in test tubes with sterile water in the dark at 24 ± 2°C. After 2 weeks, lesions were evident at the inoculation points and symptoms were similar to those caused by natural infection. P. hydropathica was consistently re-isolated from the margin of lesions, while controls remained symptomless. In the United States in 2008, P. hydropathica was described as spreading from irrigation water to Rhododendron catawbiense and Kalmia latifolia (2). This pathogen can also attack several other horticultural crops (1), but to our knowledge, this is the first report of P. hydropathica causing wilting and shoot dieback on V. tinus.

Diversity and plastid types in Dinophysis acuminata complex (Dinophyceae) in Scottish waters

Dinophysis acuminata produces lipophilic shellfish toxins (LSTs) that have economic and ecological impact on marine invertebrates in NE Atlantic where aquaculture farming is prevalent. Identification of D. acuminata can be complex. Cells exhibit a variety of morphotypes that overlap between species making identification using routine light microscopy difficult. These cells are mixotrophic and their population size is influenced by hydrographic conditions and prey populations. Dinophysis cells are able to acquire and temporarily keep prey plastids from a variety of photosynthetic unicellular sources. The Dinophysis community in Scottish waters tend to be dominated by cells with morphologies that appear to be variants of D. acuminata/norvegica complex particularly during late spring/early summer. To determine the identity of these morphotypes, DNA barcoding was performed on 32 single cell isolates from sites around the Scottish coast using the ribosomal internal transcribed spacer 1 (ITS1) and a partial cytochrome oxidase I (COI) fragment on the same single cells. Although the cells exhibited a variety of morphotypes, most were restricted to one cluster containing D. acuminata and three grouped with Dinophysis ovum. This is the first molecular confirmation of the presence of D. ovum in Scottish waters. Two isolates showed considerable divergence – one was unidentifiable from the public databases, whilst the other matched a Dinophysis cf. acuta isolate from Canada. To investigate prey plastids, molecular analysis of these Dinophysis single cells was conducted with a partial fragment of the plastid ribosomal marker (16S). Most cells harboured plastids from the cryptophyte Teleaulax – the most commonly reported plastid type, however one cell harboured a Rhodomonas/Storeatula derived plastid. This finding increases the range and variety of cryptophyte plastids found in Dinophysis and increases the range of prey-types.

Heleobia maltzani (Westerlund, 1886) (Caenogastropoda: Truncatelloidea: Cochliopidae) from Crete and species-level diversity of Heleobia Stimpson, 1865 in Europe

Morphological characters and partial cytochrome oxidase (COI) DNA sequences were used to investigate intra- and interpopulation differences among three Cretan brackish-water populations of Heleobia maltzani snails. The morphology of male and female reproductive organs of H. maltzani did not differ within studied populations or in comparison with closely related species. In contrast, analysis of shell morphology revealed substantial variation not supported by the genetic data. Genetics results showed surprisingly low differentiation between all Cretan populations and also mainland populations of Heleobia. They also showed that H. maltzani is conspecific with four other European species of Heleobia, thus implying that there is only one species of brackish-water Heleobia in Europe. Such a low level of genotypic differentiation is unusual for typical island species and is most probably the consequence of recent colonization of Crete. Possible scenarios of such processes are discussed.

Molecular and morphological variation in saddled fathead sculpins (Psychrolutidae: Cottunculus) of the north-east Atlantic Ocean, with a synonymy of three species.

On the basis of 104 individuals of Cottunculus from the Norwegian coast, the Barents Sea, Svalbard and north-east Greenland waters, this paper evaluates the descriptive taxonomic characters of three species Cottunculus sadko, Cottunculus microps and Cottunculus konstantinovi and links the character variation to genetic characterizations. Eleven morphological characters used as diagnostic characters in the original descriptions of these species were analysed in pair-wise character comparisons. All characters show cross-species traits when in combination with each other, and all individuals show traits of more than one of the species. Partial cytochrome oxidase c subunit I (coI) gene sequences from 21 individuals, covering the morphological variation among the specimens, show little variation among the individuals. From these results, it is concluded that the descriptive characters do not support the maintenance of more than one species. It is recommended that C. microps remains the name for this species, with C. sadko and C. konstantinovi as junior synonyms.

Evidence of cryptic speciation in mesostigmatid mites from South Africa.

Laelaps giganteus and Laelaps muricola (Mesostigmata; Laelapidae) are widespread and locally abundant host generalists on small mammals in southern Africa. The large host range and complex life history of these ectoparasites may allude to possible intraspecific cryptic diversity in these taxa. To assess genetic and morphological diversity in L. giganteus and L. muricola, we sampled 228 rodents at eight localities in South Africa. This sample included nine previously recorded host species and on these, L. muricola was only recorded from Mastomys natalensis and Micaelamys namaquensis while L. giganteus was found on Rhabdomys dilectus and Lemniscomys rosalia. Phylogenetic analyses of partial mtDNA cytochrome oxidase subunit I (COI) and nuclear ITS1 data strongly supported the recognition of L. giganteus and L. muricola, a scenario partly supported by the Tropomyosin intron. Strong support for evolutionary distinct lineages within L. giganteus is found: L. giganteus lineage 1 is confined to R. dilectus and L. giganteus lineage 2 is confined to L. rosalia. These host specific monophyletic lineages were also separated by 9.84% mtDNA sequence divergence and 3.44% nuclear DNA sequence divergence. Since quantitative morphometric analyses were not congruent with these findings, these two lineages more than likely represent cryptic species.

Survey on whiteflies and their parasitoids in cassava mosaic pandemic areas of Tanzania using morphological and molecular techniques.

Bemisia tabaci (Gennadius) is the vector of cassava mosaic geminiviruses (CMGs) and cassava brown streak viruses (CBSVs) in Africa, which cause devastating yield losses. As a prerequisite to developing biological control methods and enhancing knowledge of the fauna of whitefly parasitoids in sub-Saharan Africa, endemic parasitoids were surveyed in the cassava-growing regions of Tanzania and analysed using both morphological and molecular methods. An attempt was made to corroborate the identification of the parasitoid species on the basis of consideration of their morphology and sequence analyses of three DNA fragments, namely partial cytochrome oxidase I (COI), the D2 expansion segment of the 28S rRNA and the internal transcribed spacer I (ITS1). Eight whitefly species colonising cassava and twelve species of parasitoids were detected. A species in the Encarsia strenua group and a species in the Eretmocerus mundus group were the most common parasitoids. Molecular systematics indicated the occurrence of two new species of Eretmocerus Haldeman parasitising B. tabaci. The accurate identification of natural enemies is an essential first step in developing effective biological control solutions for B. tabaci in Tanzania and the wider cassava-growing environments of Africa. The new data provided here represent an important contribution to this goal.

Old Aegean palaeoevents driving the diversification of an endemic isopod species (Oniscidea, Trachelipodidae)

The Greek endemic isopod species Trachelipus aegaeus is distributed in Aegean islands and the adjacent coastal parts of the Greek mainland. Major palaeogeographic events of the Aegean archipelago, such as the formation of the mid‐Aegean trench and the Messinian Salinity Crisis, have been often employed as major causal factors of evolutionary events and phylogeographic patterns exhibited by several taxa. Herein, we infer phylogenetic relationships among T. aegaeus populations using partial cytochrome oxidase subunit I (COI) and 16S rRNA sequences. Due to the poor preservation of the specimens, we propose a modified DNA extraction protocol, which returned highly positive results in terms of the quality of the total extracted DNA. We implement a calibrated molecular clock and path sampling analysis, using alternative palaeogeographic events and rates of substitution, to evaluate the biogeographic history of the species and to estimate the chronology of diversification events among its populations. Our results are clearly in favour of the scenario of the MAT triggering vicariance among most T. aegaus populations. Moreover, the large intraspecific genetic divergence (0–19% for COI and 0–20.3% for the 16S rRNA) and the overall phylogeographic patterns depicted herein seem not to have been obscured by more recent palaeogeological events. A role of dispersal, probably human‐aided, is assumed for certain ‘deviant’ cases.