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.

Restoration of hookworm egg development after prolonged storage in stool suspension.

Hookworm infection is still prevalent in southern Thailand despite control measures. Hookworm eggs submerged for an extended period under water from rainfall or in latrines may not survive, but they may recover their ability to develop into infective larvae when exposed to atmospheric air. This study examined the survival of the hookworm eggs in stool suspension and the restoration of development capability after prolonged storage. In stool mass, eggs developed normally and yielded infective filariform larvae (FL) in 7 days. On the contrary, in 1:10 stool suspension, hookworm eggs were found to remain at the 4-8 cell stage; degenerated eggs were observed after 15 days of storage, and the number of degenerated eggs reached 80 % on day 30. Aeration of the suspension, or transferring to a Petri dish or agar plate, restored the capacity of eggs stored for up to 15 days to develop into FL; thereafter, the capacity declined sharply. Retardation of egg development under water or in stool suspension may be due to a lack of atmospheric air. Use of "night soil" from latrines as fertilizer may be one factor in maintaining hookworm transmission, as worm eggs can undergo normal development upon exposure to atmospheric air.

Effect of dilution of stool soluble component on growth and development of Strongyloides stercoralis.

Dispersion or dilution of stool by water from heavy rainfall may affect Strongyloides stercoralis free-living development producing infective filariform larvae (FL). This study examined effect of water dilution of stool on survival of S. stercoralis free-living development. One g of stool was prepared in water so that its soluble component was diluted sequentially from 1:2 to 1:480. Three dishes were used to compare FL production in three culture conditions: stool suspension, stool sediment deposited in soil, and isolated rhabditiform larvae (RhL) deposited in soil. The fourth dish was for developmental observation of RhL into free-living stages. Numerous FL were generated from undiluted or 1:2 diluted stool and stool sediment placed on soil. However, starting from dilution 1:5, FL production continuously decreased in both stool suspensions and stool sediments placed on soil. RhL isolated from stool dilutions placed on soil gave rise to few FL. Worm mating were seen at 24-30 hours in dilutions 1:20-1:120 only. Highest numbers of FL from indirect free-living cycle were 1/3 of those from control. FL production decreased as stool dilution increased, and reached zero production at 1:160 dilution. Rainfall may disperse or dilute stool so that nutritional supplement for S. stercoralis free-living development is insufficient.

The majority of cockroaches from the Samutprakarn province of Thailand are carriers of parasitic organisms.

We undertook a study of the mechanical transmission of parasitic organisms in cockroaches in the Samutprakarn province of Thailand. In this study, 920 cockroaches were obtained from 18 open-air shopping markets in 5 districts and 1 subdistrict of this province. All cockroaches were captured during their feeding time in their natural habitat. Direct wet smear and modified acid-fast bacilli staining were used to identify the parasites from the external surface or cuticle of the cockroaches. The results show that 498 (54.1 %) of the cockroaches harbored parasitic organisms. Of these, 56.1 % were protozoa and the remaining 43.9 % were helminthes (pathogenic helminthes- 1.4 %, non-pathogenic helminthes- 42.5 %). Of the pathogenic helminthes, the species included Strongyloides stercoralis (6 instances of the free-living adult male, 0.8 %), Ascaris lumbricoides (2 decorticated eggs, 0.3 %), Trichuris trichiura (2, 0.3 %), and Taenia spp. (1 egg, 0.1 %). The protozoa types that were identified included Cyclospora spp. (10 oocysts, 1.3 %), Endolimax nana (10 cysts, 1.3 %), B. hominis (9 instances of the vacuolated form, 1.2 %), Isospora belli (75 oocysts, 9.6 %), Entamoeba histolytica/E. dispar (36 cysts, 4.6 %), Cryptosporidium spp. (220 oocysts, 28.1 %), Chilomastix mesnilli (2 cysts, 0.3 %), Entamoeba coli (31 cysts, 4.0 %), Balantidium coli (45 trophozoites, 5.8 %), and Iodamoeba butschlii (1 cyst, 0.1 %). These results show that cockroaches isolated from these markets are carriers of several parasitic organisms that cause commonly found symptoms of illness such as diarrhea or bowel disorder.