Mitochondrial Genome of the Eyeworm,Thelazia callipaeda (Nematoda: Spirurida), as the First Representative from the Family Thelaziidae

Human thelaziosis is an underestimated parasitic disease caused by Thelazia species (Spirurida: Thelaziidae). The oriental eyeworm, Thelazia callipaeda, infects a range of mammalian definitive hosts, including canids, felids and humans. Although this zoonotic parasite is of socio-economic significance in Asian countries, its genetics, epidemiology and biology are poorly understood. Mitochondrial (mt) DNA is known to provide useful genetic markers to underpin fundamental investigations, but no mt genome had been characterized for any members of the family Thelaziidae. In the present study, we sequenced and characterized the mt genome of T. callipaeda. This AT-rich (74.6%) mt genome (13,668 bp) is circular and contains 12 protein-coding genes, 22 transfer RNA genes and two ribosomal RNA genes, but lacks an atp8 gene. All protein-coding genes are transcribed in the same direction; the gene order is the same as those of Dirofilaria immitis and Setaria digitata (Onchocercidae), but distinct from Dracunculus medinensis (Dracunculidae) and Heliconema longissimum (Physalopteridae). Phylogenetic analyses of the concatenated amino acid sequence data for all 12 protein-coding genes by Bayesian inference (BI) showed that T. callipaeda (Thelaziidae) is related to the family Onchocercidae. This is the first mt genome of any member of the family Thelaziidae and should represent a new source of genetic markers for studying the epidemiology, ecology, population genetics and systematics of this parasite of humans and other mammals.     

Histochemical studies on eggshell formation in Paramphistomum cervi (Digenea: Paramphistomatidae)

The formation of the egg mass in the ootype, containing the oocyte, vitelline cells, numerous spermatozoa and Mehlis' gland secretion marks the beginning of eggshell formation. Mehlis' gland cells pour their lipoprotein secretion into the ootype; this secretion forms a thin template around the egg mass and upon this template the shell globules are deposited. Eggshell formation begins in the proximal uterus and is completed in the middle uterus. The eggshells are devoid of phenols and phenolase indicating that hardening is not by quinone tanning. The eggshells contain keratin and are stabilized by disulphide linkages.     

Ultrastructure and cytochemistry of the tegument of Orthocoelium scoliocoelium and Paramphistomum cervi (Trematoda: Digenea)

The tegument of Orthocoelium scoliocoelium and Paramphistomum cervi was examined using histochemical techniques and electron microscopy. On the basis of the distribution of acid and alkaline phosphatase (E.C. 3.1.3.2, E.C. 3.1.3.1), non-specific esterase (E.C. 3.1.1.1), cholinesterase (E.C. 3.1.1.7) and succinate dehydrogenase (E.C. 1.3.99.1) at light microscope level two distinct regions were recognized, an outer and an inner zone. Electron microscopy revealed that the tegument comprises an outer surface syncytium underlain by a thick subsyncytial zone and musculature. Deeper still occur the nucleated "tegumental cells". The latter are in cytoplasmic continuity with the surface syncytium via vacuolated cytoplasmic trabeculae which traverse the muscle layers and the subsyncytial zone. Three types of tegumental cells each lacking mitochondria were observed. The T1 cells synthesize discoid and electron dense T1 bodies while T2 cells produce oval and electron lucent T2 bodies. The third type of tegumental cells apparently produce no secretory bodies and may represent an embryonic cell type. The surface syncytium contains T1 and T2 secretory bodies and is bounded apically by a plasma membrane invested externally by a fuzzy and filamentous glycocalyx. The surface syncytium lacks mitochondria and is traversed by infoldings of the basal plasma membrane. Beneath the surface syncytium the subsyncytial zone is largely comprised of fibrous interstitial material. This zone, which is particularly thick in the amphistomes, is traversed by trabeculae and extensions of underlying parenchymal cells which usually contain mitochondria and lysosomes. The subsyncytial zone overlies numerous circular and longitudinal muscle fibres. The absence of mitochondria and enzymes associated with active transport suggests that the amphistome tegument may be mainly specialized for protection of the worm against mechanical and chemical conditions prevailing in the rumen. Active uptake of nutrients is probably not a primary function.     

Morphological and histochemical observations on the vitelline cells of developing and adult Paramphistomum cervi (Trematoda: Digenea)

In 6 week-old Paramphistomum cervi, vitelline cells appear in the form of a few scattered cells, present below the body wall which divide mitotically to form vitelline follicles in 8 week-old worms. Small shell globules appear first in 10 week-old worms, the number and size of which continue to increase till the worm becomes adult. In the latter the vitelline follicles contain stem cells, immature, maturing and mature vitelline cells, which differ due to the distribution of shell globules as well as their histochemical nature. The shell globules are keratin in nature. The vitelline cells showed activities of various phosphatases but did not show the presence of any of the dehydrogenases studied.     

Morphological and histochemical studies on the prostate gland of developing and adult Paramphistomum cervi (Digenea: Paramphistomatidae)

Morphological and histochemical studies have been made on the development of the prostate gland of the digenetic trematode, Paramphistomum cervi during the course of its infection in sheep; histochemical characterization of prostate gland secretion in adult worms and its functional relationship with the transport of spermatozoa have also been studied. In 6-wk-old worms, the terminal portion of the male genital duct is formed of what appears to be a syncytial epithelium containing a relatively large number of nuclei compared to the rest of the male duct. Cellular organization of the prostate gland becomes conspicuous in 8-wk-old worms and the prostate gland is fully developed by 16 wk. Two types of prostate gland cells, characteristic of the adult, become distinct in 16-wk-old worms which contain spermatozoa in the lumen of their vas deferens and pars prostatica. Adult worms show two types of prostate gland cells: type I containing mainly glycoprotein and type II mainly phospholipid granules. Weak-to-strong activities of acid phosphatase, alkaline phosphatase, esterase, lipase, adenosine triphosphatase, tetrazolium reductase, NAD-diaphorase, isocitrate dehydrogenase, glyceraldehyde dehydrogenase, α-glycerophosphate dehydrogenase and glucose-6-phosphate dehydrogenase have been observed in the prostate gland. The release of prostate gland secretion and the passage of spermatozoa through the lumen of pars prostatica appear to be synchronized events.     

Complete Mitochondrial Genome of the Steppe Ribbon Racer(Psammophis lineolatus): The First Representative from the Snake Family Psammophiidae and its Phylogenetic Implications

Comparisons of vertebrate mitochondrial genomes(mitogenomes) may yield significant insights into the evolution of organisms and genomes. However,no complete mitogenome from the snake family Psammophiidae has been reported. In this study, we sequenced and annotated the complete mitogenome of Psammophis lineolatus, representing the first mitogenome of Psammophiidae. The total length is 17 166 bp,consisting of 13 protein-coding genes(PCGs), 22 transfer RNAs(tRNAs), two ribosomal RNAs(12 S rRNA and16 S rRNA), and duplicate control regions(CRs). This gene arrangement belongs to the Type Ⅲ pattern,which is a widely shared gene order in Alethinophidian snakes. All tRNAs exhibit cloverleaf structures with the exception of tRNA-SerAGY and tRNA-Pro, which lack a dihydrouridine(DHU) arm/stem and TΨC loop,respectively. The 13 PCGs include five start codons(ATG,GTG, ATA, ATT, and ATC), two complete stop codons(TAA and AGG), and two incomplete stop codon(T--and TA-). In addition, the Ka/Ks ratios indicate that all PCGs had undergone a strong purifying selection.Four types of CR domains rearrangement occurred among eight species of Elapoidea. The phylogenetic reconstructions with both Bayesian inference and maximum likelihood methods support the placement of Psammophiidae in the Elapoidea superfamily,with Homalopsidae being the sister taxon to Elapoidea and Colubroidea. However, the sister taxon of Psammophiidae is unclear due to the availability of Elapoidea mitogenomes being limited to the family Elapidae. More mitogenomes from different taxonomic groups in Elapoidea are needed to better understand the phylogenetic relationships within Elapoidea.     

Ultrastructure and histochemistry of the digestive tract of juvenile Paramphistomum epiclitum (Paramphistomidae: Digenea) during migration in Indian ruminants.

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and 1992. Ultrastructure and histochemistry of the digestive tract of juvenile Paramphistomum epiclitum (Paramphistomidae: Digenea) during migration in Indian ruminants. International Journal for Parasitology 22: 1089–1101. The digestive tract of juvenile Paramphistomum epiclitum consists of a foregut with a highly muscular terminal pharynx and an oesophagus, which leads to a pair of unbranched and blind-ending intestinal caeca. A syncytium lining the foregut is continuous with the external tegument and displays similar sensory papillae and secretory bodies (T1 and T2). A third type of secretory body (T3) is confined to the oesophageal cytons of newly excysted juveniles and is first evident in the syncytium by day 14 of migration. An epithelium lining the caeca is composed of a single layer of morphologically uniform cells whose apical surface is amplified by microvilli. Dense secretions synthesized in the caecal epithelium of mature cercariae are released during migration by a mechanism resembling modified apocrine discharge. The caecal epithelium of migrating juveniles undergoes a 10-fold increase in surface amplification (irrespective of growth) during its transition from a primarily secretory tissue to one apparently specialized for absorption.     

Complete Mitochondrial Genome of Echinostoma hortense (Digenea: Echinostomatidae)

Echinostoma hortense (Digenea: Echinostomatidae) is one of the intestinal flukes with medical importance in humans. However, the mitochondrial (mt) genome of this fluke has not been known yet. The present study has determined the complete mt genome sequences of E. hortense and assessed the phylogenetic relationships with other digenean species for which the complete mt genome sequences are available in GenBank using concatenated amino acid sequences inferred from 12 protein-coding genes. The mt genome of E. hortense contained 12 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and 1 non-coding region. The length of the mt genome of E. hortense was 14,994 bp, which was somewhat smaller than those of other trematode species. Phylogenetic analyses based on concatenated nucleotide sequence datasets for all 12 protein-coding genes using maximum parsimony (MP) method showed that E. hortense and Hypoderaeum conoideum gathered together, and they were closer to each other than to Fasciolidae and other echinostomatid trematodes. The availability of the complete mt genome sequences of E. hortense provides important genetic markers for diagnostics, population genetics, and evolutionary studies of digeneans.     

The First Mitochondrial Genome of the Sepsid Fly Nemopoda mamaevi Ozerov, 1997 (Diptera: Sciomyzoidea: Sepsidae), with Mitochondrial Genome Phylogeny of Cyclorrhapha

Sepsid flies (Diptera: Sepsidae) are important model insects for sexual selection research. In order to develop mitochondrial (mt) genome data for this significant group, we sequenced the first complete mt genome of the sepsid fly Nemopoda mamaevi Ozerov, 1997. The circular 15,878 bp mt genome is typical of Diptera, containing all 37 genes usually present in bilaterian animals. We discovered inaccurate annotations of fly mt genomes previously deposited on GenBank and thus re-annotated all published mt genomes of Cyclorrhapha. These re-annotations were based on comparative analysis of homologous genes, and provide a statistical analysis of start and stop codon positions. We further detected two 18 bp of conserved intergenic sequences from tRNA^Glu-tRNA^Phe and ND1-tRNA^Ser(UCN) across Cyclorrhapha, which are the mtTERM binding site motifs. Additionally, we compared automated annotation software MITOS with hand annotation method. Phylogenetic trees based on the mt genome data from Cyclorrhapha were inferred by Maximum-likelihood and Bayesian methods, strongly supported a close relationship between Sepsidae and the Tephritoidea.     

Ultrastructure and histochemistry of the protonephridial system of juvenile Paramphistomum epiclitum and Fischoederius elongatus (Paramphistomidae: Digenea) during migration in Indian ruminants.

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and 1992. Ultrastructure and histochemistry of the protonephridial system of juvenile Paramphistomum epiclitum and Fischoederius elongatus (Paramphistomidae: Digenea) during migration in Indian ruminants. International Journal for Parasitology 22: 1103–1115. The protonephridial system of juvenile Paramphistomum epiclitum and Fischoederius elongatus consists of a bilaterally symmetrical arrangement of primary, secondary and tertiary ducts which connect individual flame cells with a simple common bladder. Primary and secondary ducts are formed from columns of adjoining cells which provide an epithelial lining, whose luminal surface is elaborated with either short tubercles or lamellae. Groups of cilia project from the luminal surface at frequent intervals along secondary ducts. By contrast, the tertiary ducts and bladder are lined with a nucleated syncytium which ends at a junctional complex formed with the terminal canal. The latter is continuous with the tegumental syncytium and opens at a nephridiopore on the postero-dorsal surface. Tertiary ducts of mature cercariae contain concretions which are voided by migrating juveniles in whose tertiary ducts lipids are progressively accumulated. Evidence for the role of protonephridia in excretion and possibly in osmoregulation and ionic balance is currently examined.     

The First Complete Mitochondrial Genome Sequence in the Genus Aphanius (Teleostei)

Journal of Ichthyology - In the present study, for the first time we isolated and characterized the complete mitochondrial genome sequence of the endangered Farsi killifish Aphanius farsicus by...     

The First Mitochondrial Genome for Caddisfly (Insecta: Trichoptera) with Phylogenetic Implications

The Trichoptera (caddisflies) is a holometabolous insect order with 14,300 described species forming the second most species-rich monophyletic group of animals in freshwater. Hitherto, there is no mitochondrial genome reported of this order. Herein, we describe the complete mitochondrial (mt) genome of a caddisfly species, Eubasilissa regina (McLachlan, 1871). A phylogenomic analysis was carried out based on the mt genomic sequences of 13 mt protein coding genes (PCGs) and two rRNA genes of 24 species belonging to eight holometabolous orders. Both maximum likelihood and Bayesian inference analyses highly support the sister relationship between Trichoptera and Lepidoptera.     

The First Mitochondrial Genome for the Fishfly Subfamily Chauliodinae and Implications for the Higher Phylogeny of Megaloptera

Megaloptera are a basal holometabolous insect order with larvae exclusively predacious and aquatic. The evolutionary history of Megaloptera attracts great interest because of its antiquity and important systematic status in Holometabola. However, due to the difficulties identifying morphological apomorphies for the group, controversial hypotheses on the monophyly and higher phylogeny of Megaloptera have been proposed. Herein, we describe the complete mitochondrial (mt) genome of a fishfly species, Neochauliodes punctatolosus Liu & Yang, 2006, representing the first mt genome of the subfamily Chauliodinae. A phylogenomic analysis was carried out based on the mt genomic sequences of 13 mt protein-coding genes (PCGs) and two rRNA genes of nine Neuropterida species, comprising all three orders of Neuropterida and all families and subfamilies of Megaloptera. Both maximum likelihood and Bayesian inference analyses highly support the monophyly of Megaloptera, which was recovered as the sister of Neuroptera. Within Megaloptera, the sister relationship between Corydalinae and Chauliodinae was corroborated. The divergence time estimation suggests that stem lineage of Neuropterida and Coleoptera separated in the Early Permian. The interordinal divergence within Neuropterida might have occurred in the Late Permian.     

The First Mitochondrial Genome for the Superfamily Hagloidea and Implications for Its Systematic Status in Ensifera

Hagloidea Handlirsch, 1906 was an ancient group of Ensifera, that was much more diverse in the past extending at least into the Triassic, apparently diminishing in diversity through the Cretaceous, and now only represented by a few extant species. In this paper, we report the complete mitochondrial genome (mitogenome) of Tarragoilus diuturnus Gorochov, 2001, representing the first mitogenome of the superfamily Hagloidea. The size of the entire mitogenome of T. diuturnus is 16144 bp, containing 13 protein-coding genes (PCGs), 2 ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes and one control region. The order and orientation of the gene arrangement pattern is identical to that of D. yakuba and most ensiferans species. A phylogenomic analysis was carried out based on the concatenated dataset of 13 PCGs and 2 rRNA genes from mitogenome sequences of 15 ensiferan species, comprising four superfamilies Grylloidea, Tettigonioidae, Rhaphidophoroidea and Hagloidea. Both maximum likelihood and Bayesian inference analyses strongly support Hagloidea T. diuturnus and Rhaphidophoroidea Troglophilus neglectus as forming a monophyletic group, sister to the Tettigonioidea. The relationships among four superfamilies of Ensifera were (Grylloidea, (Tettigonioidea, (Hagloidea, Rhaphidophoroidea))).     

Effect of temperature on the infectivity of metacercariae of Zygocotyle lunata (Digenea: Paramphistomidae)

As a test of the energy limitation hypothesis (ELH), we predicted that temperature would have a significant influence on the infectivity of metacercariae of the digenetic trematode Zygocotyle lunata. Snails infected with Z. lunata were collected from ponds near Crawfordsville, Indiana, isolated at room temperature, and examined for the release of cercariae. Newly encysted metacercariae were collected and incubated 1-30 days at 1 of 5 temperatures (0, 3, 25, 31, 37 C). Twenty-five cysts were fed to each of 5 or 10 mice per treatment group (temperature). At 17 days postinfection, mice were killed and worms were recovered; data were collected on levels of infection in each group and the total body area of each worm. No worms were found in mice fed cysts that had been held at 0 C or 37 C (after 30 days). There were no differences in prevalence, infectivity, or mean intensity among the 3, 25, and 31 C treatments. Infectivity of metacercariae incubated at 37 C for 1 day was significantly greater than in all other treatments, while infectivity of metacercariae in the 37 C/15-day treatment was significantly lower than in all others. Mean body area of worms at 37 C/15 days was significantly greater than at other temperatures, suggesting density-dependent increases in growth. These results, particularly those from the 37 C treatments, are consistent with the ELH; infectivity was lower at high temperatures or when incubated for more time at 1 temperature (fewer energy reserves). It is suggested that microhabitat conditions experienced by metacercariae of Z. lunata could contribute to longer larval life, thus influencing this parasite's temporal dispersal.     

Ultrastructural and cytochemical studies on the ventral pouch of Gastrothylax crumenifer (Digenea:Paramphistomidae)

The ventral pouch of G. crumenifer consists of an extensive atrium, opening just behind the mouth and ending blindly at the level of the testes. It is lined by a thin tegument underlain by numerous flask shaped pyriform cells. The non-glandular nature of the pyriform cell cytoplasm and the presence of haemoglobin and numerous mitochondria suggest that the pouch fulfils primarily a respiratory role. Contractions and relaxations of musculature associated with the pouch may ventilate the pouch with ruminal fluid, or with gaseous oxygen which may be present in small quantities in the rumen. Oxygen taken up by haemoglobin in pyriform cells may be transferred to haemoglobin in the lymph system, and thence distributed internally.