Ultrastructure and localization of Neorickettsia in adult digenean trematodes provides novel insights into helminth-endobacteria interaction

Abstract Background Neorickettsia are a group of intracellular α proteobacteria transmitted by digeneans (Platyhelminthes, Trematoda). These endobacteria can also infect vertebrate hosts of the helminths and cause serious diseases in animals and humans. Neorickettsia have been isolated from infected animals and maintained in cell cultures, and their morphology in mammalian cells has been described. However, limited information is available on the morphology and localization of Neorickettsia in the trematode host. Methods We used a Neorickettsia-infected strain of the model trematode Plagiorchis elegans to infect Syrian Golden hamsters to produce adult worms. Ultrastructure of Neorickettsia was assessed by transmission electron microscopy of high pressure freezing/freeze substitution fixed specimens. A Neorickettsia surface protein from P. elegans (PeNsp-3) was cloned and antibodies against the recombinant protein were used to localize Neorickettsia by immunohistochemistry. Results Ultrastructural analysis revealed moderate numbers of pleomorphic endobacteria with a median size of 600 × 400 nm and characteristic double membranes in various tissue types. Endobacteria showed tubular membrane invaginations and secretion of polymorphic vesicles. Endobacteria were unevenly localized as single cells, or less frequently as small morula-like clusters in the ovary, Mehlis’ gland, vitelline follicles, uterus, intrauterine eggs, testis, cirrus-sac, tegument, intestine and the oral and ventral sucker. Examination of hamster small intestine infected with P. elegans showed many endobacteria at the host-parasite interface such as the oral and ventral sucker, the tegument and the excretory pore. Conclusions We conclude that adult P. elegans trematodes carry Neorickettsia endobacteria in varying numbers in many tissue types that support vertical transmission, trematode to trematode transmission via seminal fluid, and possibly horizontal transmission from trematodes to vertebrate hosts. These means appear to be novel mechanisms of pathogen transmission by endoparasitic worms

Two New Species of \u3ci\u3eRhabdias\u3c/i\u3e (Nematoda: Rhabdiasidae) from the Marine Toad, \u3ci\u3eBufo marinus\u3c/i\u3e (L.) (Lissamphibia: Anura: Bufonidae), in Central America

Two new Rhabdias species are described from the lungs of the cane toad Bufo marinus (L.) from Costa Rica and Nicaragua. Rhabdias alabialis n. sp. differs from other known species of the genus by the remarkable morphology of its head end, i.e., the absence of lips or pseudolabia, the slitlike oral opening, and the triangular shape of the buccal capsule in apical view. Rhabdias pseudosphaerocephala n. sp. is identified as a form previously known in Central and South America as Rhabdias sphaerocephala Goodey, 1924, a species initially described from toads in Europe. The new species is differentiated from R. sphaerocephala based on head-end morphology and sequences of nuclear rDNA

Two New Species of \u3ci\u3eRhabdias\u3c/i\u3e (Nematoda: Rhabdiasidae) from the Marine Toad, \u3ci\u3eBufo marinus\u3c/i\u3e (L.) (Lissamphibia: Anura: Bufonidae), in Central America

Two new Rhabdias species are described from the lungs of the cane toad Bufo marinus (L.) from Costa Rica and Nicaragua. Rhabdias alabialis n. sp. differs from other known species of the genus by the remarkable morphology of its head end, i.e., the absence of lips or pseudolabia, the slitlike oral opening, and the triangular shape of the buccal capsule in apical view. Rhabdias pseudosphaerocephala n. sp. is identified as a form previously known in Central and South America as Rhabdias sphaerocephala Goodey, 1924, a species initially described from toads in Europe. The new species is differentiated from R. sphaerocephala based on head-end morphology and sequences of nuclear rDNA

A Review of \u3ci\u3ePolylekithum Arnold\u3c/i\u3e, 1934 and Its Familial Affinities Using Morphological and Molecular Data, With Description of \u3ci\u3ePolylekithum catahoulensis\u3c/i\u3e sp nov.

The type material of Polylekithum ictaluri, P. halli, and Maculifer chandleri was examined from the United States National Parasite Museum, and we determined that the material was conspecific, making P. halli and M. chandleri junior subjective synonyms of P. ictaluri. Polylekithum catahoulensis sp. nov. was described from material collected from catfishes at the Catahoula Wildlife Refuge, LaSalle Parish, Louisiana, USA, and compared with P. ictaluri collected from catfishes in Reelfoot Lake, Obion County, Tennessee, USA, and the Pearl River, Hancock County, Mississippi, USA. Polylekithum catahoulensis had smaller eggs (77–88 μm long by 51–63 μm wide vs. 94–108 μm by 52–76 μm) and a longer forebody (35–41% of overall body length vs. 29–34%). Comparison of more than 2,400 bp long fragments of nuclear ribosomal DNA (complete ITS and partial 28S regions) strongly supported the status of P. catahoulensis as a new species. Molecular phylogenetic analysis of 28S rDNA gene sequences from Polylekithum as well as representative species from Allocreadiidae, Atractotrematidae, Brachycoeliidae, Callodistomidae, Dicrocoeliidae, Encyclometridae, Gorgoderidae, Haploporidae, Opecoelidae, Plagiorchiidae, and Telorchiidae rooted by Monorchiidae and Lissorchiidae demonstrated that of the families tested, Polylekithum was most closely related to Encyclometridae as a gorgoderoid and not to Allocreadiidae as previously reported. Morphological features of three South American allocreadiids, Allocreadium patagonicum, P. percai, and A. pichi were inconsistent with generic diagnoses of Allocreadium and Polylekithum, so we suggested they belonged in a single unnamed genus similar to Creptotrema. Polylekithum catlai from India was assessed from the description and failed to conform to the generic diagnosis of Polylekithum. Morphology of Caudouterina suggested a close relationship with Polylekithum and not Allocreadiidae

The Innovative Approaches in the Staff Management of Enterprises

The article is aimed at studying and systematizing the basic modern theoretical approaches to innovative staff management. The most important innovative approaches in staff management of enterprises presented in the scientific-methodological researches of contemporary scholars are considered and defined. The necessity of scientific substantiation of introduction of innovations in the sphere of staff management by definition of key principles of innovative development of this sphere is identified. The authors also consider the main traditional (administrative, economic, social-psychological) and the innovative methods (innovative selection of staff, providing the use of the newest technologies of search for employees; innovative methods of assessment of activities used at the stage of recruitment and management of personnel to determine the level of conformity of both professional and personal characteristics of staff with both official and internal requirements; innovative career management methods; innovative methods of stimulation and motivation) in the system of modern staff management of enterprises, the most priority aspects of their introduction and implementation are allocated

Large Scale Screening of Digeneans for \u3cem\u3eNeorickettsia\u3c/em\u3e Endosymbionts Using Real-Time PCR Reveals New \u3cem\u3eNeorickettsia\u3c/em\u3e Genotypes, Host Associations and Geographic Records

Digeneans are endoparasitic flatworms with complex life cycles including one or two intermediate hosts (first of which is always a mollusk) and a vertebrate definitive host. Digeneans may harbor intracellular endosymbiotic bacteria belonging to the genus Neorickettsia (order Rickettsiales, family Anaplasmataceae). Some Neorickettsia are able to invade cells of the digenean\u27s vertebrate host and are known to cause diseases of wildlife and humans. In this study we report the results of screening 771 digenean samples for Neorickettsia collected from various vertebrates in terrestrial, freshwater, brackish, and marine habitats in the United States, China and Australia. Neorickettsia were detected using a newly designed real-time PCR protocol targeting a 152 bp fragment of the heat shock protein coding gene, GroEL, and verified with nested PCR and sequencing of a 1371 bp long region of 16S rRNA. Eight isolates of Neorickettsia have been obtained. Sequence comparison and phylogenetic analysis demonstrated that 7 of these isolates, provisionally named Neorickettsia sp. 1–7 (obtained from allocreadiid Crepidostomum affine, haploporids Saccocoelioides beauforti and Saccocoelioides lizae, faustulid Bacciger sprenti, deropegid Deropegus aspina, a lecithodendriid, and a pleurogenid) represent new genotypes and one (obtained from Metagonimoides oregonensis) was identical to a published sequence of Neorickettsia known as SF agent. All digenean species reported in this study represent new host records. Three of the 6 digenean families (Haploporidae, Pleurogenidae, and Faustulidae) are also reported for the first time as hosts of Neorickettsia. We have detected Neorickettsia in digeneans from China and Australia for the first time based on PCR and sequencing evidence. Our findings suggest that further surveys from broader geographic regions and wider selection of digenean taxa are likely to reveal new Neorickettsia lineages as well as new digenean host associations

Molecular phylogeny provides new insights on the taxonomy and composition of Lyperosomum Looss, 1899 (Digenea, Dicrocoeliidae) and related genera

Lyperosomum Looss, 1899 is one of the largest genera of the Dicrocoeliidae and is one of the best examples of the systematic complexity and taxonomic instability within this family. We present the molecular analyses based on novel sequences of nuclear and mitochondrial genes obtained from 56 isolates of adult flukes and larval stages of dicrocoeliids belonging to Lyperosomum, Skrjabinus, Zonorchis as well as previously available sequence data. According to obtained results we propose to return Zonorchis clathratus and Z. petiolatus into Lyperosomum, and to recognize L. alagesi as a synonym of L. petiolatum. Our study shows that L. petiolatum commonly occurs in Europe in corvids as well as in several species of migratory songbirds, e.g. Sylvia atricapilla. At the same time, the Turdidae appear to host a distinct species of Lyperosomum. The phylogenetic analysis has clearly demonstrated the paraphylepic nature of Lyperosomum and indicated the need of its thorough revision preferably using specimens from type hosts and type territories of nominal species. In addition, inclusion of numerous not yet sequenced dicrocoeliid genera into future phylogenetic studies is necessary to clarify the interrelationships of taxa within the family and stabilize its system

A descrição matemática do desempenho de sensores e biossensores eletroquímicos de ph, baseados em polímeros condutores, nos processos de medição do ph dos líquidos de cavidade bucal no modo de voltagem constante

O desempenho de sensores e biossensores de pH, baseados nos polímeros condutores, usados no modo de voltagem constante para medir o pH da cavidade bucal, foi descrito matematicamente. Por meio da análise do respetivo modelo, foram inferidas as condições da melhor precisão do indicador, da multiplicidade de estado estacionário e do comportamento oscilatório em parâmetro eletroquímico

Comparing the mitochondrial genomes of Wolbachia-dependent and independent filarial nematode species

BACKGROUND: Many species of filarial nematodes depend on Wolbachia endobacteria to carry out their life cycle. Other species are naturally Wolbachia-free. The biological mechanisms underpinning Wolbachia-dependence and independence in filarial nematodes are not known. Previous studies have indicated that Wolbachia have an impact on mitochondrial gene expression, which may suggest a role in energy metabolism. If Wolbachia can supplement host energy metabolism, reduced mitochondrial function in infected filarial species may account for Wolbachia-dependence. Wolbachia also have a strong influence on mitochondrial evolution due to vertical co-transmission. This could drive alterations in mitochondrial genome sequence in infected species. Comparisons between the mitochondrial genome sequences of Wolbachia-dependent and independent filarial worms may reveal differences indicative of altered mitochondrial function. RESULTS: The mitochondrial genomes of 5 species of filarial nematodes, Acanthocheilonema viteae, Chandlerella quiscali, Loa loa, Onchocerca flexuosa, and Wuchereria bancrofti, were sequenced, annotated and compared with available mitochondrial genome sequences from Brugia malayi, Dirofilaria immitis, Onchocerca volvulus and Setaria digitata. B. malayi, D. immitis, O. volvulus and W. bancrofti are Wolbachia-dependent while A. viteae, C. quiscali, L. loa, O. flexuosa and S. digitata are Wolbachia-free. The 9 mitochondrial genomes were similar in size and AT content and encoded the same 12 protein-coding genes, 22 tRNAs and 2 rRNAs. Synteny was perfectly preserved in all species except C. quiscali, which had a different order for 5 tRNA genes. Protein-coding genes were expressed at the RNA level in all examined species. In phylogenetic trees based on mitochondrial protein-coding sequences, species did not cluster according to Wolbachia dependence. CONCLUSIONS: Thus far, no discernable differences were detected between the mitochondrial genome sequences of Wolbachia-dependent and independent species. Additional research will be needed to determine whether mitochondria from Wolbachia-dependent filarial species show reduced function in comparison to the mitochondria of Wolbachia-independent species despite their sequence-level similarities