Correction: Genetic diversity of Entamoeba: Novel ribosomal lineages from cockroaches.

[This corrects the article DOI: 10.1371/journal.pone.0185233.].

Genetic diversity of Entamoeba: Novel ribosomal lineages from cockroaches.

Our current taxonomic perspective on Entamoeba is largely based on small-subunit ribosomal RNA genes (SSU rDNA) from Entamoeba species identified in vertebrate hosts with minor exceptions such as E. moshkovskii from sewage water and E. marina from marine sediment. Other Entamoeba species have also been morphologically identified and described from non-vertebrate species such as insects; however, their genetic diversity remains unknown. In order to further disclose the diversity of the genus, we investigated Entamoeba spp. in the intestines of three cockroach species: Periplaneta americana, Blaptica dubia, and Gromphadorhina oblongonota. We obtained 134 Entamoeba SSU rDNA sequences from 186 cockroaches by direct nested PCR using the DNA extracts of intestines from cockroaches, followed by scrutinized BLASTn screening and phylogenetic analyses. All the sequences identified in this study were distinct from those reported from known Entamoeba species, and considered as novel Entamoeba ribosomal lineages. Furthermore, they were positioned at the base of the clade of known Entamoeba species and displayed remarkable degree of genetic diversity comprising nine major groups in the three cockroach species. This is the first report of the diversity of SSU rDNA sequences from Entamoeba in non-vertebrate host species, and should help to understand the genetic diversity of the genus Entamoeba.

Identification of Entamoeba polecki with Unique 18S rRNA Gene Sequences from Celebes Crested Macaques and Pigs in Tangkoko Nature Reserve, North Sulawesi, Indonesia.

Unique species of macaques are distributed across Sulawesi Island, Indonesia, and the details of Entamoeba infections in these macaques are unknown. A total of 77 stool samples from Celebes crested macaques (Macaca nigra) and 14 stool samples from pigs were collected in Tangkoko Nature Reserve, North Sulawesi, and the prevalence of Entamoeba infection was examined by PCR. Entamoeba polecki was detected in 97% of the macaques and all of the pigs, but no other Entamoeba species were found. The nucleotide sequence of the 18S rRNA gene in E. polecki from M. nigra was unique and showed highest similarity with E. polecki subtype (ST) 4. This is the first case of identification of E. polecki ST4 from wild nonhuman primates. The sequence of the 18S rRNA gene in E. polecki from pigs was also unique and showed highest similarity with E. polecki ST1. These results suggest that the diversity of the 18S rRNA gene in E. polecki is associated with differences in host species and geographic localization, and that there has been no transmission of E. polecki between macaques and pigs in the study area.

Giardia lamblia transcriptome analysis using TSS-Seq and RNA-Seq.

Giardia lamblia is a protozoan parasite that is found worldwide and has both medical and veterinary importance. We applied the transcription start sequence (TSS-seq) and RNA sequence (RNA-seq) techniques to study the transcriptome of the assemblage A WB strain trophozoite. We identified 8000 transcription regions (TR) with significant transcription. Of these regions, 1881 TRs were more than 500 nucleotides upstream of an annotated ORF. Combining both techniques helped us to identify 24 ORFs that should be re-annotated and 60 new ORFs. From the 8000 TRs, we were able to identify an AT-rich consensus that includes the transcription initiation site. It is possible that transcription that was previously thought to be bidirectional is actually unidirectional.

Contribution of the Japan International Cooperation Agency health-related projects to health system strengthening.

BACKGROUND: The Japan International Cooperation Agency (JICA) has focused its attention on appraising health development assistance projects and redirecting efforts towards health system strengthening. This study aimed to describe the type of project and targets of interest, and assess the contribution of JICA health-related projects to strengthening health systems worldwide. METHODS: We collected a web-based Project Design Matrix (PDM) of 105 JICA projects implemented between January 2005 and December 2009. We developed an analytical matrix based on the World Health Organization (WHO) health system framework to examine the PDM data and thereby assess the projects' contributions to health system strengthening. RESULTS: The majority of JICA projects had prioritized workforce development, and improvements in governance and service delivery. Conversely, there was little assistance for finance or medical product development. The vast majority (87.6%) of JICA projects addressed public health issues, for example programs to improve maternal and child health, and the prevention and treatment of infectious diseases such as AIDS, tuberculosis and malaria. Nearly 90% of JICA technical healthcare assistance directly focused on improving governance as the most critical means of accomplishing its goals. CONCLUSIONS: Our study confirmed that JICA projects met the goals of bilateral cooperation by developing workforce capacity and governance. Nevertheless, our findings suggest that JICA assistance could be used to support financial aspects of healthcare systems, which is an area of increasing concern. We also showed that the analytical matrix methodology is an effective means of examining the component of health system strengthening to which the activity and output of a project contributes. This may help policy makers and practitioners focus future projects on priority areas.

Full-parasites: database of full-length cDNAs of apicomplexa parasites, 2010 update.

Full-Parasites (http://fullmal.hgc.jp/) is a transcriptome database of apicomplexa parasites, which include Plasmodium and Toxoplasma species. The latest version of Full-Parasites contains a total of 105,786 EST sequences from 12 parasites, of which 5925 full-length cDNAs have been completely sequenced. Full-Parasites also contain more than 30 million transcription start sites (TSS) for Plasmodium falciparum (Pf) and Toxoplasma gondii (Tg), which were identified using our novel oligo-capping-based protocol. Various types of cDNA data resources were interconnected with our original database functionalities. Specifically, in this update, we have included two unique RNA-Seq data sets consisting of 730 million mapped RNA-Seq tags. One is a dataset of 16 time-lapse experiments of cultured bradyzoite differentiation for Tg. The other dataset includes 31 clinical samples of Pf. Parasite RNA was extracted together with host human RNA, and the extracted mixed RNA was used for RNA sequencing, with the expectation that gene expression information from the host and parasite would be simultaneously represented. By providing the largest unique full-length cDNA and dynamic transcriptome data, Full-Parasites is useful for understanding host-parasite interactions and will help to eventually elucidate how monophyletic organisms have evolved to become parasites by adopting complex life cycles.

Characterization of alpha-phosphoglucomutase isozymes from Toxoplasma gondii.

The Toxoplasma gondii genome project has revealed two putative isoforms (TgPGM-I and TgPGM-II) of alpha-phosphoglucomutase (EC 5.4.2.2). We obtained recombinant proteins of these isoforms from the Beverley strain of T. gondii and characterized their properties, particularly the kinetic properties of these isoforms. The specific activities of TgPGM-I and TgPGM-II for alpha-D-glucose 1-phosphate were 338+/-9 and 84+/-6micromol/min/mg protein, respectively, at 37 degrees C under optimal conditions. The Kcat and Km values of TgPGM-I were 398+/-11/s and 0.19+/-0.03mM and those for TgPGM-II were 93+/-7/s and 3.53+/-0.91mM, respectively, for alpha-d-glucose 1-phosphate. Magnesium ions were the most effective divalent cations for both the enzyme activities. The maximum activities of both the enzymes were obtained in the presence of more than 0.2mM alpha-D-glucose 1,6-bisphosphate. Although both enzymes were attached to the alpha-phosphohexomutase superfamily, amino acid sequence homology between TgPGM-I and TgPGM-II showed very low overall identity (25%). No alpha-phosphomannomutase (EC 5.4.2.8) activity was detected for either enzyme. The data indicated that TgPGM-I, but not TgPGM-II, may play an important role in alpha-D-glucose 6-phosphate production.

The high prevalence of asymptomatic Toxocara infection among schoolchildren in Manado, Indonesia.

We performed a serological survey of Toxocara canis infection in junior high school students from three districts in northern Sulawesi. Almost all of the 117 subjects from two rural districts near Manado allowed dogs in their houses, and there was an 84.6% prevalence of T. canis infection in this group. Fifty-three subjects (45.3%) had serum samples with a high titer of specific anti-Toxocara antibody. By contrast, 41 students tested in one urban district showed a 12.2% prevalence. To confirm the clinical symptoms of visceral larva migrans (VML) and ocular larva migrans (OLM) caused by Toxocara, we administered a questionnaire survey, serological liver function tests, and an ophthalmoscopic examination in 34 subjects having high anti-Toxocara antibodies. One rural district showed a high prevalence; 58 out of 71 subjects (81.7%) had a high titer of anti-Toxocara antibodies according to a plate-ELISA test, although none showed clinical signs. Five of these subjects exhibited hypereosinophilia. These results indicated that T. canis infection in northern Sulawesi is latent in many more cases than previously estimated, and suggest that people living in environments polluted by Toxocara eggs become easily infected with T. canis and show a high prevalence of infection.