Diagnostic and analytical mutation scanning of Cryptosporidium: utility and advantages.

Cryptosporidiosis is predominantly a disease of the alimentary tract of humans and other vertebrates, caused by parasitic protists of the genus Cryptosporidium. This disease, transmitted mainly via the fecal-oral route (in water or food), is of major socioeconomic importance globally. The diagnosis of cryptosporidiosis, including the genetic characterization of the different species, genotypes and subgenotypes (population variants) of Cryptosporidium, is crucial to prevention and control, particularly as there is no cost-effective treatment against this disease.

Redescription and genetic characterization of selected Contracaecum spp. (Nematoda: Anisakidae) from various hosts in Australia.

In the present study, five species of Contracaecum Railliet & Henry 1912 (Nematoda: Ascaridida), including Contracaecum bancrofti, Contracaecum microcephalum, Contracaecum variegatum, Contracaecum eudyptulae, and Contracaecum ogmorhini, were redescribed using light and scanning electron microscopy. In addition, in order to elucidate their taxonomic status, first and second internal transcribed spacers (ITS-1 and ITS-2, respectively) of nuclear ribosomal DNA of each morphospecies were characterized. Analyses of sequence and morphological data sets suggested that C.

The mitochondrial genomes of Ancylostoma caninum and Bunostomum phlebotomum--two hookworms of animal health and zoonotic importance.

Background: Hookworms are blood-feeding nematodes that parasitize the small intestines of many mammals, including humans and cattle. These nematodes are of major socioeconomic importance and cause disease, mainly as a consequence of anaemia (particularly in children or young animals), resulting in impaired development and sometimes deaths. Studying genetic variability within and among hookworm populations is central to addressing epidemiological and ecological questions, thus assisting in the control of hookworm disease.

Systematic analysis of insertions and deletions specific to nematode proteins and their proposed functional and evolutionary relevance.

Background: Amino acid insertions and deletions in proteins are considered relatively rare events, and their associations with the evolution and adaptation of organisms are not yet understood. In this study, we undertook a systematic analysis of over 214,000 polypeptides from 32 nematode species and identified insertions and deletions unique to nematode proteins in more than 1000 families and provided indirect evidence that these alterations are linked to the evolution and adaptation of nematodes.

Results: Amino acid alterations in sequences of nematodes were identified by comparison with homologous sequences from a wide range of eukaryotic (metzoan) organisms.

A polymerase chain reaction-coupled high-resolution melting curve analytical approach for the monitoring of monospecificity of avian Eimeria species.

Coccidiosis is a significant disease of poultry caused by different species of Eimeria. Differentiation of Eimeria species is important for the quality control of the live attenuated Eimeria vaccines derived from monospecific lines of Eimeria spp. In this study, high-resolution melting (HRM) curve analysis of the amplicons generated from the second internal transcribed spacer of nuclear ribosomal DNA (ITS-2) was used to distinguish between seven pathogenic Eimeria species of chickens, and the results were compared with those obtained from the previously described technique, capillary electrophoresis.

Improved molecular diagnostic tools for human hookworms.

Blood-feeding hookworms are parasitic roundworms (i.e., nematodes) of major socioeconomic importance, affecting millions of people worldwide.

Investigation of the regulation of transcriptional changes in Ancylostoma caninum larvae following serum activation, with a focus on the insulin-like signalling pathway.

The exit from dauer in the free-living nematode Caenorhabditis elegans is under the control of a single amphidial neuron (ASJ) of the insulin-like signalling pathway. Mutations of this pathway have the ability to suppress entry into the dauer stage. It has been postulated that insulin-like signalling plays a significant role in the response to serum stimulation in vitro of the third-stage larvae (L3s) of the canine hookworm Ancylostoma caninum.

High resolution melting-curve (HRM) analysis for the diagnosis of cryptosporidiosis in humans.

Cryptosporidiosis of humans is an intestinal disease caused predominantly by infection with Cryptosporidium hominis or C. parvum. This disease is transmitted mainly via the faecal-oral route (water or food) and has major socioeconomic impact globally.

Analysis of the genetic diversity within Cryptosporidium hominis and Cryptosporidium parvum from imported and autochtonous cases of human cryptosporidiosis by mutation scanning.

The present study investigated sequence variation in part of the 60 kilodalton glycoprotein (pgp60) gene among Cryptosporidium hominis and Cryptosporidium parvum isolates (n=115) from citizens of the UK inferred to have been infected whilst travelling abroad (to 25 countries) or in the UK. The genomic DNA samples from these isolates were subjected to PCR-coupled single-strand conformation polymorphism analysis, followed by targeted sequencing of pgp60. Individual samples were classified to the genotypic and subgenotypic levels based on phylogenetic analysis (Bayesian inference) of pgp60 data, including published sequences for comparison.

Improved insights into the transcriptomes of the human hookworm Necator americanus--fundamental and biotechnological implications.

Hookworms of humans are blood-feeding parasitic nematodes of major socio-economic significance in a wide range of countries. They cause a neglected tropical disease (NTD) called "hookworm disease" (=necatoriasis and/or ancylostomiasis). Necator americanus is the most widely distributed hookworm of humans and is a leading cause of iron deficiency anaemia, which can cause physical and mental retardation and deaths in children as well as adverse maternal-foetal outcomes.

Needles in the EST haystack: large-scale identification and analysis of excretory-secretory (ES) proteins in parasitic nematodes using expressed sequence tags (ESTs).

Background: Parasitic nematodes of humans, other animals and plants continue to impose a significant public health and economic burden worldwide, due to the diseases they cause. Promising antiparasitic drug and vaccine candidates have been discovered from excreted or secreted (ES) proteins released from the parasite and exposed to the immune system of the host. Mining the entire expressed sequence tag (EST) data available from parasitic nematodes represents an approach to discover such ES targets.

Tv-RIO1 - an atypical protein kinase from the parasitic nematode Trichostrongylus vitrinus.

Background: Protein kinases are key enzymes that regulate a wide range of cellular processes, including cell-cycle progression, transcription, DNA replication and metabolic functions. These enzymes catalyse the transfer of phosphates to serine, threonine and tyrosine residues, thus playing functional roles in reversible protein phosphorylation. There are two main groups, namely eukaryotic protein kinases (ePKs) and atypical protein kinases (aPKs); RIO kinases belong to the latter group.

Genetic divergence between island and continental populations of the parasitic nematode Labiosimplex australis in Australia.

Sequences of the first and second internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal (r) DNA were characterised for Labiosimplex australis, a parasitic nematode of macropodid marsupials from continental Australia and from island populations which have been isolated from mainland Australia for relatively short periods of time (<40,000 years). The results showed that the geographically isolated populations of L. australis on Kangaroo Island and Tasmania were genetically different from each other and from populations on the mainland.

Contracaecum pyripapillatum n. sp. (Nematoda: Anisakidae) and a description of C. multipapillatum (von Drasche, 1882) from the Australian pelican, Pelecanus conspicillatus.

A new species of nematode, Contracaecum pyripapillatum, is reported from the Australian pelican, Pelecanus conspicillatus. This species resembles Contracaecum multipapillatum which was found in the same host. These two species can be differentiated from one another based on the shape of the preanal papillae, being pyriform in C.

The mitochondrial genome of Toxocara canis.

Toxocara canis (Ascaridida: Nematoda), which parasitizes (at the adult stage) the small intestine of canids, can be transmitted to a range of other mammals, including humans, and can cause the disease toxocariasis. Despite its significance as a pathogen, the genetics, epidemiology and biology of this parasite remain poorly understood. In addition, the zoonotic potential of related species of Toxocara, such as T.

Genomic-bioinformatic analysis of transcripts enriched in the third-stage larva of the parasitic nematode Ascaris suum.

Differential transcription in Ascaris suum was investigated using a genomic-bioinformatic approach. A cDNA archive enriched for molecules in the infective third-stage larva (L3) of A. suum was constructed by suppressive-subtractive hybridization (SSH), and a subset of cDNAs from 3075 clones subjected to microarray analysis using cDNA probes derived from RNA from different developmental stages of A.

Extensive and complex sequence diversity in mitochondrial cytochrome c oxidase subunit 1 within Necator americanus from Colombia revealed by SSCP-coupled sequencing.

In spite of the human health importance of hookworms, there has been only a small number of studies of genetic variability within Necator americanus, and none in South America. In the present study, we investigated sequence variability in a 395-bp region of the mitochondrial cytochrome c oxidase subunit 1 gene among N. americanus individuals (n=100) from humans of six villages in Colombia, employing a mutation scanning-coupled sequencing approach.

Assessing the relationship between Malassezia and leishmaniasis in dogs with or without skin lesions.

The relationship among the frequency, population size and phospholipase activity of Malasseziapachydermatis was investigated for dogs with Leishmania infantum infection (Li+) and those without evidence of this infection (Li-). A group of 188 dogs (141 without and 47 with skin lesions) was examined clinically, and samples were taken for the detection of Malassezia and L. infantum using various diagnostic methods.

Toward practical, DNA-based diagnostic methods for parasitic nematodes of livestock--bionomic and biotechnological implications.

Parasitic nematodes of livestock have a major economic impact worldwide. In spite of the health problems caused by nematodes and advances toward the development of vaccines and new therapeutic agents against some of them, relatively limited attention has been paid to the need for improved, practical methods of diagnosis. Accurate diagnosis and genetic characterization of parasitic nematodes of livestock are central to their effective control, particularly given the current, serious problems with anthelmintic resistance in nematode populations.

Classification of Cryptosporidium species from patients with sporadic cryptosporidiosis by use of sequence-based multilocus analysis following mutation scanning.

In the present study, we analyzed genetic variation in Cryptosporidium species from humans (n = 62) with clinical cryptosporidiosis in South Australia. Sequence variation was assessed in regions within the small subunit of nuclear rRNA (p-SSU), the 70-kDa heat shock protein (p-hsp70), and the 60-kDa glycoprotein (p-gp60) genes by employing single-strand conformation polymorphism analysis and sequencing. Based on the analyses of p-SSU and p-hsp70, Cryptosporidium hominis (n = 38) and Cryptosporidium parvum (n = 24) were identified.

In silico analysis of expressed sequence tags from Trichostrongylus vitrinus (Nematoda): comparison of the automated ESTExplorer workflow platform with conventional database searches.

Background: The analysis of expressed sequence tags (EST) offers a rapid and cost effective approach to elucidate the transcriptome of an organism, but requires several computational methods for assembly and annotation. Researchers frequently analyse each step manually, which is laborious and time consuming. We have recently developed ESTExplorer, a semi-automated computational workflow system, in order to achieve the rapid analysis of EST datasets.

Genetic variants of Malassezia pachydermatis from canine skin: body distribution and phospholipase activity.

Malassezia pachydermatis isolates (n=185) from skin sites from dogs (n=30) were characterized genetically and biochemically following in vitro culture. Two regions in the chitin synthase-2 gene (chs-2) and the first internal transcribed spacer (ITS-1) of nuclear ribosomal DNA were sequenced, and the phospholipase activity of each isolate was assessed. Three chs-2 (i.

Parasite transmission by insects: a female affair?

Understanding the relationship between the gender of insects and their ability to act as vectors of insect-borne diseases (IBDs) could provide clues as to the origin of the intimate interplay among insect, pathogen and vertebrate hosts. The vector activity of several species of blood-feeding insects is linked to adult females. Interestingly, the only known exception is the transmission of canine and human thelaziosis by a male dipteran fly.

Transcriptional changes in the hookworm, Ancylostoma caninum, during the transition from a free-living to a parasitic larva.

Background: Third-stage larvae (L3) of the canine hookworm, Ancylostoma caninum, undergo arrested development preceding transmission to a host. Many of the mRNAs up-regulated at this stage are likely to encode proteins that facilitate the transition from a free-living to a parasitic larva. The initial phase of mammalian host invasion by A.