Partial 16S rDNA Research Articles

Morphological, histopathological and molecular characterization of Myxobolus szekelyianus n. sp. (Cnidaria: Myxosporea: Myxobolidae) causing acute gill disease in Schizothorax esocinus (Heckel, 1838) from River Jhelum of Kashmir Himalayan region, India

During the present study, we describe a novel myxozoan that forms plasmodia in the gill filament of Schizothorax esocinus collected from River Jhelum of Kashmir Himalayan region (n = 26). Description of the new species is based on morphology of the myxospore and on partial 18S rDNA gene. The results showed that the plasmodia were large, visible with naked eye, elongate in shape, whitish in colour, 8mm in diameter, at the centre of gill filament, histozoic, single or two in number per gill, but too big in size and filled by 1,000–1,400 myxospores. The myxospores measure 13.4 (length) × 9.5µm (width), short oval in frontal view, blunt at both in anterior and posterior, but somewhat narrowing towards the posterior end. Shell valves measure 0.45µm in thickness. No parietal folds were recorded. Two polar capsules equal in size, pyriform in shape were measured 7.3 × 4.2 µm (L × W). The 18S rDNA of this species (accession number MK785257) clustered phylogenetically with other related myxozoan parasites and showed maximum homogeneity of 98.63% with M. sp. SKBU-RC1 (MK412937) and M. sp. SKBU-RC2 (MK412937) infecting gills of Labeo rohita from India followed by 92.51% with M. pronini (MH329619) infecting abdominal cavity of Carassius gibelio from China. The study of histopathological changes of the gills revealed hypertrophy, hyperplasia, uplifting of cellular elements and severe damage to gill filaments resulting in suffocation of fish. By its unique myxospore morphology and its 18S rDNA sequence, we propose naming this species as Myxobolus szekelyianus n. sp.

The first case of Spiroxys contortus in European pond turtle (Emys orbicularis) in the wild in Poland.

During standard parasitological dissection of 4 individuals of European pond turtle (E. orbicularis) (Linneaus, 1758) one nematode was found. In a morphological examination the parasite was identified as Spiroxys contortus (Rudolphi, 1819) and confirmed by DNA analysis based on sequencing of the small subunit ribosomal RNA gene. The partial 18S rDNA gene was deposited to NCBI GenBank with the accession number MN629917. This is the first molecular evidence of S. contortus in E. orbicularis from wild from Poland. The analyzed sample genotype sequence shows 100% similarity to the reference specimen from Argentina.

Hepatozoon cevapii (Apicomplexa: Hepatozoidae) in the Thamnodynastes lanei snake (Colubridae, Tachymenini) from the Eastern Amazon, Brazil.

The aim of the present study was to verify the occurrence of hemogregarines in the colubrid snake Thamnodynastes lanei from the eastern Amazon region of Brazil. Intraerythrocytic gamonts with mean dimensions of 14.8 ± 1.8 × 4.0 ± 0.7μm and encapsulated gamonts with mean dimensions of 15.3 ± 1.1 × 4.8 ± 0.5μm were observed. Through morphological and molecular data based on the partial 18S rDNA gene, the parasite was identified as Hepatozoon cevapii, originally described in the viperid snake Crotalus durissus terrificus from the southeast region of Brazil. Thus, the findings of the present study extend the geographic range of H. cevapii and provide novel Hepatozoon-snake associations.

Description of Ditylenchus acantholimonis n. sp. (Rhabditida: Anguinidae) from Iran, a morphological and molecular phylogenetic study

Summary Ditylenchus acantholimonis n. sp. is described based on morphological, morphometric and molecular characters. It was isolated from the rhizosphere soil of Acantholimon sp. in Golestan province, Iran, and is mainly characterised by having four lines in the lateral field, a pyriform to bottle-shaped offset pharyngeal bulb, post-vulval uterine sac 36.6-56.1% of the vulva to anus distance long, and a subcylindrical to conical tail with widely rounded tip. It is further characterised by short to medium-sized females, 480-617 μm long, with a fine stylet having small rounded knobs, V = 80.8-83.6, c = 11.0-13.8, c′ = 3.3-4.6, and males with 16.0-17.0 μm long spicules. The new species was morphologically compared with six species having four lines in their lateral field, rounded tail tip and comparable morphometric data namely: D. dipsacoideus, D. emus, D. exilis, D. paraparvus, D. sturhani, and D. solani. It was also compared with two species, D. ferepolitor and D. angustus, forming a maximally supported clade in the 18S tree. The phylogenetic analyses using the maximal number of Anguinidae and several Sphaerularioidea genera based upon partial 18S and 28S rDNA D2-D3 sequences revealed that Ditylenchus is polyphyletic. In the 18S tree, the new species formed a clade with D. ferepolitor (KJ636374) and D. angustus (AJ966483); in the 28S tree it formed a poorly supported clade with D. phyllobios (KT192618) and Ditylenchus sp. (MG865719).

Morphological and molecular characterization of Ameloblastella pirarara sp. n. (Monogenoidea: Dactylogyridae) parasitizing the large Amazonian catfish Phractocephalus hemioliopterus

In this study, integrative taxonomy is applied to describe a new dactylogyrid species, Ameloblastella pirarara sp. n. from the gills of Phractocephalus hemioliopterus, a commercially and ecologically important Amazonian catfish. Ameloblastella pirarara sp. n. can be distinguished from its congeners mainly by the morphology of the male copulatory organ (MCO), accessory piece, and anchors. The new species most resembles Ameloblastella unapi, from the Peruvian Amazon, but differs from it by the number of MCO rings, morphology of the vaginal canal and sclerotized structures of the haptor. Phylogenetic analyses based on sequences of the partial 28S rDNA (D1-D2 domains) gene placed the new species in a well-supported subclade of Ameloblastella spp. parasites of Neotropical siluriform fish, as a sister taxon to Ameloblastella unapioides. Thus, the new species described herein expands our knowledge of the diversity of monogenoid parasites from Amazonian freshwater fish.

Comparative and phylogenomic analysis of nuclear and organelle genes in cryptic Coelastrella vacuolata MACC-549 green algae

The nuclear, chloroplast and mitochondrial genomes of a unicellular green algal species of the Coelastrella genus was sequenced, assembled and annotated. The strain was previously classified as Chlamydomonas sp. MACC-549 based on morphology and partial 18S rDNA analysis. However, the proposed multi-loci phylogenomic approach described in this paper placed this strain within the Coelastrella genus, therefore it was re-named to Coelastrella vacuolata MACC-549. The strain was selected for de novo sequencing based on its potential value in biohydrogen production as revealed in earlier studies. This is the first thorough report and characterization for green algae from the Coelastrella genus. The whole genome annotation of Coelastrella vacuolata MACC-549 (including nuclear, chloroplast and mitochondrial genomes) shed light on interesting metabolic and sexual breeding features of this algae and served as a basis to taxonomically classify this strain.

Phylogenetics and Pathogenicity of Balantioides coli Isolates in Vietnamese Weaned Pigs.

This study aimed to investigate the diversity of Balantioides coli(B. coli) based on the 18S rDNA of isolates collected from weaned pigs with diarrhea and then select a B.coliisolate to determine its pathogenicity in weaned pigs through experimental infection. The genetic analysis of field-isolated B. coli strains was based on sequencing of the partial 18S rDNA genetic fragment and the pathogenicity of an isolate of B. coli (DN2018-2-DTD) was identified through an experimental infection. The fourteen B.coliisolates shared 98.47-100% nucleotide similarity and were divided into two clades in a phylogenetic tree. In an infection experiment, a slight increase in the body temperature was identified in two infected groups as compared to that in the control group (P < 0.05) from days post-infection (dpi) 12-18 and from dpi 20-27. In the group infected with 3000 CPG/TPG, 2/9 pigs started to develop diarrhea on day two. In the group infected with 500 CPG/TPG, 8/9 pigs had diarrhea on dpi 3. Trophozoites/cysts of B.coliwere firstly detected in feces on dpi 3 in a group of pigs infected with a dose of 3000 CPG/TPG, and CPG/TPG reached its peak at dpi 5 in both groups of infected pigs (35,450 and 13,250 CPG/TPG). There was a significant difference in the bodyweight of the control group (noninfected) as compared to that of the groups of infected pigs on dpi 7 (P < 0.05), 14 (P < 0.001) and 28 (P < 0.01). Lesions were solely recorded in infected pigs, including white ulcerative nodules with necrosis scattered in the mucosa of the colon and cecum on dpi 7, 14, and 28. This is the first study to investigate the pathogenicity of B.coliin Vietnamese pigs to demonstrate that B.colicould induce necrotic ulcerative enteritis and diarrhea in weaned pigs.

Biodiversity, phylogeny and toxin production profile of cyanobacterial strains isolated from lake Latyan in Iran

Monitoring toxigenic cyanobacteria in freshwaters is of great importance due to the adverse health impacts on humans and aquatic organisms. Here we studied cyanobacterial occurrence and biodiversity in a drinking water reservoir in Tehran province, Iran. In total, nine different species representing three orders of Synechococcales, Oscillatoriales and Nostocales were isolated and classified into six families and seven genera ranging from 92.3% to 99.0% similarities in their partial 16S rDNA with GenBank sequences. The cultures were analyzed for cyanotoxins production by the Artemia salina bioassay, ultrahigh performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and also screened for the presence of marker genes involved in toxins production. Ethyl acetate extracts of three strains showed more than 50% mortality on A. salina larvae after 24 h at a concentration of 500 µg/ml. Production of at least one of the cyanotoxins, microcystin (MC), cylindrospermopsin (CYN) and anatoxin-a (ATX-a), was detected in 6 of the strains. Seven MC variants with a total concentration of 130.6 ng/mg of biomass dry weight were detected for the strain Phormidium sp. UTMC6001 and molecular screening of the mcyE gene also confirmed the presence of this biomarker in its genome. Our study also revealed the production of CYN in a novel picocyanobacterial strain Cyanobium sp. UTMC6007 at 1.0 ng/mg of biomass dry weight. Considering the limited information on freshwater toxic cyanobacteria taxonomy in the Middle East, these findings will expand our knowledge and consequently aid in development of new water management policies in future.

Bio-plastic Films Production from Feather Waste Degradation by Keratinolytic Bacteria Bacillus cereus

Plastic materials have become a necessity of human life especially in the packaging of food commodities and biomedical procedures. Bioplastic is emerging as an effective alternative to fossil oil-based materials to avoid the environmental hazards of the plastic industry. During this study, chicken feathers were used as a substrate to isolate keratin degrading bacteria. Among 14 identified isolates, Bacillus sp BAM3 was found to be the most promising isolate. Partial 16S rDNA analysis-based molecular characterization revealed it is a strain of Bacillus cereus. Bacillus sp BAM3 can grow and produce keratinase in feathers containing basal medium as the sole carbon and energy source. The maximum keratinase production (730U/ml) was achieved within 24 h under optimum reaction conditions. The optimized reaction pH and temperature were noted as 9.0 and 50 °C for crude keratinase activity, respectively. The chicken feathers were used as a substrate in 2, 5, and 10 wt% glycerol to synthesize keratin-based bioplastic with keratinolytic bacterium Bacillus cereus BAM3. Bioplastic prepared from keratin with 2% of glycerol was found to possess good mechanical properties. Therefore, the results present a novel keratinolytic isolate of Bacillus cereus BAM3, which may have potential biotechnological applications in keratin hydrolysis processes. The development of keratin-based bioplastics possessing superior crystalline morphology requires further investigations to substitute fossil oil-based materials.

Morphological and molecular characterization of Udonella brasiliensis n. sp. (Monogenoidea), an epibiont on Caligus sp. parasite of Ariidae from the southeastern coast of Brazil.

The present study describes Udonella brasiliensis n. sp., an epibiont found on Caligus sp., a parasite the ariids Genidens barbus (Lacepède) and Aspistor luniscutis (Valenciennes), caught on the coast of the state of São Paulo, Brazil. Morphological and molecular analyses (partial 18S rDNA) were carried out. The morphological data showed that U. brasiliensis n. sp. can be distinguished from current valid species by its morphometric attributes (e.g., body, pharynx, ovary and testis), while the molecular information supports the proposal of a new species. The 18S rDNA phylogenetic analysis shows a close relationship between the new species and Udonella australis Carvajal & Sepulveda, in a subclade formed of species that parasitize South American fish. Finally, this study also discusses a scenario of initial irradiation for udonellids.

First report on genetic characterization, cell-surface properties and pathogenicity of Lactococcus garvieae, emerging pathogen isolated from cage-cultured cobia (Rachycentron canadum).

The diseased cage-cultured cobia (Rachycentron canadum) displayed clinical signs, haemorrhagic eyes, dorsal darkness and gross pathological lesions, enlargement of spleen and liver. Haemorrhages were found in brain, heart and liver with cumulative mortality rates ranging from 20% to 50%. Extensive congestion in the heart, liver, spleen, kidney and brain was observed histopathologically. Epicarditis and meningitis were also revealed in diseased cobia. All isolates recovered from the organs (liver, spleen, head kidney, posterior kidney, brain and muscle) of cobia were found to be gram-positive, non-motile, ovoid cocci, short-chain-forming (diplococci) and α-haemolytic. The API 32 strep system together with the polymerase chain reaction assay for species-specific primers (pLG1 and pLG2) and the internal transcribed spacer (ITS) region (G1 and L1 primers) confirmed all four selected isolates as Lactococcus garvieae. Partial 16S rDNA nucleotide sequence (~1,100bp) of one representative L.garvieae isolate AOD109191 (GenBank accession number, MW328528.1) shared 99.9% identities with the 16S rDNA nucleotide sequence of L.garvieae (GenBank accession numbers: MT604790.1). Transmission electron microscopy (TEM) evaluation of one representative L.garvieae isolate (AOD109191) and the results of multiplex PCR did not reveal the presence of the capsular gene cluster (CGC), thus categorizing the isolate as the KG+ phenotype. Capsule staining and TEM observations confirmed the presence of a hyaluronic acid-like capsule, a possible virulence factor in KG+ phenotype L.garvieae isolates. The pathogenic potential of the representative isolate (AOD109191) was assessed through intraperitoneal injection challenges in cobia. The gross lesions and histopathological changes found in experimentally infected cobia were similar to those seen in naturally infected fish. This is the first report that confirms L.garvieae-induced 'warm water lactococcsis' can cause outbreaks of diseases in cage-cultured cobia.

Redescription and New Host Record of Heteraxinoides atlanticus (Monogenea: Heteraxinidae) from the Gills of Nemipterus japonicus (Bloch) and Its Systematics

Background: Heteraxinoides atlanticus Gayevskaya et Kovaleva (1979) is described from the locality of Arabian Sea, India. This monogenean species is first time reported from India, infesting the gills of Nemipterus japonicus (Bloch). The present study redescribes H. atlanticus and its phylogenetic status using morphometric and molecular tools. Methods: A total of 35 parasites were collected from the 234 specimens of Nemipterus japonicus at Mumbai, India. The temporary and permanent mounts were prepared for the morphometric analyses of H. atlanticus. For PCR, the genomic DNA was isolated from the parasites using primers for 18S rDNA, 28S rDNA and mtCOI gene. The obtained DNA sequences were subjected to different bioinformatics softwares (BLASTn, Clustal Omega and MEGA) for phlyogenetic analyses.Result: Phylogenetic analyses with the help of partial 18S and 28S rDNA sequences of H. atlanticus and other available sequences of heteraxinids provided the better understanding of relationship in the family Heteraxinidae. Another species, H. karachiensis Hadi and Bilqees (2014) showed similar morphological features such as number of clamps and structure of genital atrium to H. atlanticus except few minor differences, so it must be considered as synonym of H. atlanticus and placed into Heteraxinidae instead of Axinidae.

Rediscovery and first genetic description of some poorly known tick species: Haemaphysalis kopetdaghica Kerbabaev, 1962 and Dermacentor raskemensis Pomerantzev, 1946

This study aimed to provide novel information for some poorly known/rare tick species collected from wild goats (Capra aegagrus) in the mountains of Eastern Anatolia, Turkey and to expand upon the available genetic data. The collected ticks were morphologically identified as Haemaphysalis kopetdaghica (all active stages, n = 140), Dermacentor raskemensis (adults, n = 7), Ixodes gibbosus (adults, n = 15), Rhipicephalus kohlsi (female, n = 1), and R. bursa (nymphs, n = 2). A total of 32 engorged ticks (6 larvae, 6 nymphs, and 20 females) collected were allowed to molt to the next stage or for egg laying and larval hatching, respectively. In addition, one R. kohlsi female (previously confirmed by SEM microscopy) collected from a wild goat in the neighboring province of Erzurum was included in this study for further genetic comparison. The partial mitochondrial 16S rDNA and cytochrome c oxidase subunit 1 (barcoding regions) genes of each tick species were sequenced. All DNA samples obtained from the ticks were checked by PCR for the presence of Anaplasma spp., Babesia spp., Borrelia burgdorferi sensu lato, spotted fever group rickettsiae, and Theileria spp., but were found to be negative. Phylogenetic analyses of the 16S rDNA and cox1 genes were performed using the ML method to determine their genetic relationship with related ticks. As a result, this study has: i) rediscovered and provided two new tick records (H. kopetdaghica and D. raskemensis) for Turkey, ii) provided the first genetic data for H. kopetdaghica and D. raskemensis and revealed their phylogenetic relationships, iii) characterized the cox1 region of I. gibbosus for the first time, and iv) revealed significant genetic diversity between R. kohlsi from Anatolia and R. kohlsi from Oman, suggesting that R. kohlsi could include a cryptic species.

Campylobacter vulpis sp. nov. isolated from wild red foxes

During a sampling of wild red foxes (Vulpes vulpes) for the detection of Epsilonproteobacteria, 14 strains were isolated from the caecal contents of 14 epidemiologically-unrelated animals. A genus-specific PCR indicated that the isolates belonged to the genus Campylobacter. Based on the results of a species-specific PCR, the isolates were initially identified as C. upsaliensis. However, multi-locus sequence typing (MLST) revealed that the isolates were significantly different from the C. upsaliensis present in the MLST database. A polyphasic study, including conventional biochemical and tolerance characteristics, morphology by transmission electron microscopy (TEM), MALDI-TOF analysis, and genetic comparisons based on partial 16S rDNA and atpA gene sequences, was undertaken. Finally, the complete genome sequence of the type strain 251/13T and the draft genome sequences of the other isolates were determined. Average nucleotide identity, average amino acid identity and in silico DNA-DNA hybridization analyses confirmed that the isolates represent a novel taxon for which the name Campylobacter vulpis sp. nov. is proposed, with isolate 251/13T (=CCUG 70587T = LMG 30110T) as the type strain. In order to allow a rapid discrimination of C. vulpis from the closely-related C. upsaliensis, a specific PCR test was designed, based on atpA gene sequences.

Redescription and molecular characterization of Baruscapillaria spiculata (Nematoda: Capillariidae) parasitizing the Neotropic cormorant Phalacrocorax brasilianus from two Argentinian lagoons.

Two species of intestinal Capillariidae were hitherto known from the Neotropic cormorant Phalacrocorax brasilianus, Baruscapillaria spiculata (Freitas, 1933), and Baruscapillaria appendiculata (Freitas, 1933). The original descriptions are very short and brief, and further reports of both species are scarce and/or confusing. This paper provides a morphological redescription and molecular characterization, based on the partial 18S rDNA gene, of B. spiculata specimens parasitizing the Neotropic cormorant in two continental lagoons from Buenos Aires province, Argentina. Both morphological and morphometrical differences between B. spiculata and B. appendiculata are highlighted on the examination of available type material. Additionally, two previous reports of B. appendiculata from Mexico and Brazil are discussed. A phylogenetic analysis conducted on specimens of B. spiculata and 46 other capillariid isolates available from the GenBank demonstrated a sister-taxon relationship between our specimens and the type species of Baruscapillaria. But, at the same time, significant genetic distances between both taxa showed an interesting variability of the genus Baruscapillaria. The probable division of this genus into multiple genera could probably be confirmed through integrative studies including more species.

Against growing synonymy: Identification pitfalls of Hepatozoon and Schellackia demonstrated on North Iranian reptiles

The analyses of molecular data represent an effective tool for increasing the credibility of taxonomy and facilitate the description of species. Nevertheless, in haemoprotozoa, the growing amount of available sequential data is not matched by the still limited number of well-defined species. We identified four protistan haemoparasites in North Iranian reptiles: two Hepatozoon and two Schellackia species. Hepatozoon colubri and Hepatozoon ophisauri were morphologically identified in their type hosts, their partial 18S rDNA was analyzed, and thorough literature data were included in their redescription. The scarce data on the detected Schellackia spp. did not allow for their formal species description. Using an integrative approach, including morphological and geographical features, host specificity, molecular data, and the data published thus far, we face the following main difficulties hindering reliable diagnosis. (1) The lack of molecular data on well-described and named species. (2) The insufficiency of using only morphological and biological features, or only sequential data without morphology, to perform an absolutely reliable species diagnosis. (3) Typical morphological features are more substantial than metric means. (4) High risk of synonymy is present in taxonomy of blood Protista. (5) Artefacts caused by blood smear processing further complicate the correct morphological determination.

Molecular identification of a novel intracellular proteobacteria from scallop Chlamys farreri

The scallop Chlamys farreri is an important bivalve species cultured in the north of China. With the large expansion of culturing, mass mortalities have occurred persistently and caused great economic loss since 1996. An intracellular Rickettsiales-like prokaryote (RLP) was proposed as the causative agent. To identify the RLP, a partial 16S rDNA sequence of 1319 base pairs was amplified, cloned and sequenced. Phylogenetic analysis indicates that the prokaryote is a member of the α subclass of the proteobacteria. In the phylogenetic tree, it forms an independent and novel clade, which shares an evolutionary line of descent with a group of uniquely obligate intracellular organism comprised of Caedibacter caryophilus, an endosymbiont of Acanthamoeba sp., and the necrotizing hepatopancreatitis (NHP) bacterium of shrimp. Based on the obtained 16S rDNA sequence, three specific RNA targeted oligonucleotide DNA probes were designed, synthesized and labeled with digoxigenin for in situ hybridization detection. Results showed that the probes specifically bonded to the RLP inclusion in infected C. farreri tissues, indicating the amplified, cloned and sequenced 16S rDNA originated from the intracellular RLP parasitizing the C. farreri. We propose the name Sinorickettsia chlamys sp.nov. to C. farreri RLP.

New insights on the diversity of Brazilian anuran blood parasites: With the description of three new species of Hepatozoon (Apicomplexa: Hepatozoidae) from Leptodactylidae anurans.

The Amphibia are considered the most threatened vertebrate class globally, yet in Brazil they are also one of the more diverse and species rich groups. Although, in recent years there has been strong focus on amphibian related research, their parasites have not received the same attention. In Brazil, only a single species of Hepatozoon, namely H. leptodactyli (Lesage, 1908) Pessoa, 1970, has been described from anuran hosts. The present study aimed to describe three new species of Hepatozoon parasitising Leptodactylus labyrinthicus and Leptodactylus latrans from Mato Grosso State, Brazil. From 66 anurans screened for haemogregarines, four belonging to the Leptodactylidae were found positive for species of Hepatozoon. Based on the morphological analysis of peripheral blood gamonts and spleen and liver tissue meronts, three different morphotypes of Hepatozoon spp. were identified. Morphotype 1 (M1) and morphotype 2 (M2) in L. labyrinthicus and morphotype 3 (M3) in L. latrans. Molecular data based on partial 18S rDNA sequences revealed an interspecific divergence, between the species ranging from 0.43% to 1.16%. Phylogenetic analysis recovered isolates from the present study monophyletic with other isolates from Brazilian reptile and anuran hosts, sister to a clade comprising species isolated from African, North American and European reptile and anuran host species. Thus, using morphological and molecular analysis three new species infecting Brazilian Leptodactylidae anurans were identified and described. This study increases the knowledge of Brazilian anurans blood parasites and demonstrates the importance of using integrative approaches for diagnosis of hemoparasites.

The molecular profile of Acanthogyrus (Acanthosentis) kashmirensis from the Indian subcontinent.

The purpose of the present study was to provide molecular support for the validity of the morphological description of Acanthosentis kashmirensis Amin, Heckmann, Zargar, 2017 which was originally poorly described as Neoechinorhynchus kashmirensis Fotedar and Dar, 1977 from the Indian subcontinent, and to characterise its molecular identity and phylogenetic relationships. Total DNA was extracted, and the partial region of the small subunit (SSU) 18S rDNA and ITS-rDNA genes were amplified and sequenced. Genetic diversity was calculated and phylogenetic analysis of the nucleotide sequence data was performed. In this study, the molecular profile of this acanthocephalan was generated for the first time. Based on the partial 18S rDNA, interspecific variation between A. kashmirensis with different species of Acanthosentis and were 3.6-10.3% and 22.4-38.2% based on 18S rDNA and ITS-rDNA genes, respectively. We described the phylogenetic relationships of A. kashmirensis compared with other species of the genus and also with members of the family Quadrigyridae. The ITS-rDNA sequences of members of the family Quadrigyridae are more variable than 18S rDNA that can be useful for achieving a proper assessment of biodiversity. Sequence data generation from additional species of Acanthosentis will be needed to reconstruct the phylogenetic relationships of this group of acanthocephalans.

Elastic Light Scatter Pattern Analysis for the Expedited Detection of Yersinia Species in Pork Mince: Proof of Concept.

Isolation of the pathogens Yersinia enterocolitica and Yersinia pseudotuberculosis from foods typically rely on slow (10–21 day) “cold enrichment” protocols before confirmed results are obtained. We describe an approach that yields results in 39 h that combines an alternative enrichment method with culture on a non-selective medium, and subsequent identification of suspect colonies using elastic light scatter (ELS) analysis. A prototype database of ELS profiles from five Yersinia species and six other bacterial genera found in pork mince was established, and used to compare similar profiles of colonies obtained from enrichment cultures from pork mince samples seeded with representative strains of Y. enterocolitica and Y. pseudotuberculosis. The presumptive identification by ELS using computerised or visual analyses of 83/90 colonies in these experiments as the target species was confirmed by partial 16S rDNA sequencing. In addition to seeded cultures, our method recovered two naturally occurring Yersinia strains. Our results indicate that modified enrichment combined with ELS is a promising new approach for expedited detection of foodborne pathogenic yersiniae.