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Il Presidente, la Vicepresidente, i membri degli Organi Sociali e i collaboratori della Fondazione Cassa di Risparmio di Bolzano partecipano commossi al dolore della famiglia e dei parenti tutti per la scomparsa del
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Il dott. Spögler è stato il primo Presidente della Fondazione Cassa di Risparmio di Bolzano. Nel Suo periodo di attività, tra il 1992 e il 1994, ha dato contenuto, forma ed indirizzo all’attività del nostro ente. Il suo fattivo apporto lascerà profonde e sostenibili impronte.

A Lui serberemo un perenne ricordo.

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Emerg Infect Dis

v.18(9); 2012 Sep

PMC3437720

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Emerg Infect Dis. 2012 Sep; 18(9): 1543–1545.
University of Virginia School of Medicine, Charlottesville, Virginia, USA (T.L. Royer, C. Gilchrist, K.S. Ralston, W.A. Petri, Jr.);
International Centre for Diarrheal Disease Research, Dhaka, Bangladesh (M. Kabir, T. Arju, R. Haque);
and London School of Hygiene and Tropical Medicine, London, UK (C.G. Clark)
Address for correspondence: William A. Petri, Jr., Division of Infectious Diseases and International Health, University of Virginia Health System, Carter Harrison Bldg, Rm 1709A, 345 Crispell Dr, PO Box 801340, Charlottesville, VA 22908-1340, USA; email: ude.ainigriv@g3paw
Keywords: Entamoeba, Entamoeba Bangladeshi, polymerase chain reaction, phylogeny, ribosomal RNA, Bangladesh, parasites
This is a publication of the U.S. Government. This publication is in the public domain and is therefore without copyright. All text from this work may be reprinted freely. Use of these materials should be properly cited.
Technical Appendix Sequencing outcomes for 13 samples that were negative by PCR for Entamoeba histolytica , E. dispar , and E. moshkovskii and taxonomic summary for E. Bangladeshi .
GUID: 8752D7F0-3D9F-4635-9B05-9578EC32FFB8
Oligonucleotide primers used for screening and sequencing of Entamoeba bangladeshi nov. sp., Bangladesh*
1. Mondal D, Haque R, Sack RB, Kirkpatrick BD, Petri WA Jr. Attribution of malnutrition to cause-specific diarrheal illness: evidence from a prospective study of preschool children in Mirpur, Dhaka, Bangladesh.
Am J Trop Med Hyg . 2009; 80 :824–6. [ PMC free article ] [ PubMed ] [ Google Scholar ]
2. Shimokawa C, Kabir M, Taniuchi M, Mondal M, Kobayashi S, Ali IK, et al.
Entamoeba moshkovskii is associated with diarrhea in infants and causes diarrhea and colitis in mice. J Infect Dis. 2012. Jun 21; [Epub ahead of print]. [ PMC free article ] [ PubMed ]
3. Haque R, Mondal D, Karim A, Molla IH, Rahim A, Faruque ASG, et al.
Prospective case-control study of the association between common enteric protozoal parasites and diarrhea in Bangladesh.
Clin Infect Dis . 2009; 48 :1191–7. 10.1086/597580 [ PMC free article ] [ PubMed ] [ CrossRef ] [ Google Scholar ]
4. Petri WA Jr, Haque R. Entamoeba species, including Amebiasis. In: Mandell GL, JE Bennett, R Dolin, eds. Mandell, Douglas, and Bennett's principles and practice of infectious diseases, 7th ed. Philadelphia: Churchill Livingstone; 2010. p. 3411–25. [ Google Scholar ]
5. Ali IKM, Zaki M, Clark CG. Use of PCR amplification of tRNA gene-linked short tandem repeats for genotyping Entamoeba histolytica.
J Clin Microbiol . 2005; 43 :5842–7. 10.1128/JCM.43.12.5842-5847.2005 [ PMC free article ] [ PubMed ] [ CrossRef ] [ Google Scholar ]
6. Stensvold CR, Lebbad M, Victory EL, Verweij JJ, Tannich E, Alfellani M, et al.
Increased sampling reveals novel lineages of Entamoeba : consequences of genetic diversity and host specificity for taxonomy and molecular detection.
Protist . 2011; 162 :525–41. 10.1016/j.protis.2010.11.002 [ PubMed ] [ CrossRef ] [ Google Scholar ]
7. Ali IKM, Hossain MB, Roy S, Ayeh-Kumi PF, Petri WA, Haque R, et al.
Entamoeba moshkovskii infections in children, Bangladesh.
Emerg Infect Dis . 2003; 9 :580–4. 10.3201/eid0905.020548 [ PMC free article ] [ PubMed ] [ CrossRef ] [ Google Scholar ]
8. Haque R, Kabir M, Noor Z, Rahman SMM, Mondal D, Alam F, et al.
Diagnosis of amebic liver abscess and amebic colitis by detection of Entamoeba histolytica DNA in blood, urine, and saliva by a real-time PCR assay.
J Clin Microbiol . 2010; 48 :2798–801. 10.1128/JCM.00152-10 [ PMC free article ] [ PubMed ] [ CrossRef ] [ Google Scholar ]
9. Haque R, Ali IK, Akther S, Petri WA Jr. Comparison of PCR, isoenzyme analysis, and antigen detection for diagnosis of Entamoeba histolytica infection.
J Clin Microbiol . 1998; 36 :449–52. [ PMC free article ] [ PubMed ] [ Google Scholar ]
10. Haque R, Faruque AS, Hahn P, Lyerly DM, Petri WA. Entamoeba histolytica and Entamoeba dispar infection in children in Bangladesh.
J Infect Dis . 1997; 175 :734–6. 10.1093/infdis/175.3.734 [ PubMed ] [ CrossRef ] [ Google Scholar ]
Articles from Emerging Infectious Diseases are provided here courtesy of Centers for Disease Control and Prevention
1. Mondal D, Haque R, Sack RB, Kirkpatrick BD, Petri WA Jr. Attribution of malnutrition to cause-specific diarrheal illness: evidence from a prospective study of preschool children in Mirpur, Dhaka, Bangladesh.
Am J Trop Med Hyg . 2009; 80 :824–6. [ PMC free article ] [ PubMed ] [ Google Scholar ] [ Ref list ]
2. Shimokawa C, Kabir M, Taniuchi M, Mondal M, Kobayashi S, Ali IK, et al.
Entamoeba moshkovskii is associated with diarrhea in infants and causes diarrhea and colitis in mice. J Infect Dis. 2012. Jun 21; [Epub ahead of print]. [ PMC free article ] [ PubMed ] [ Ref list ]
3. Haque R, Mondal D, Karim A, Molla IH, Rahim A, Faruque ASG, et al.
Prospective case-control study of the association between common enteric protozoal parasites and diarrhea in Bangladesh.
Clin Infect Dis . 2009; 48 :1191–7. 10.1086/597580 [ PMC free article ] [ PubMed ] [ CrossRef ] [ Google Scholar ] [ Ref list ]
4. Petri WA Jr, Haque R. Entamoeba species, including Amebiasis. In: Mandell GL, JE Bennett, R Dolin, eds. Mandell, Douglas, and Bennett's principles and practice of infectious diseases, 7th ed. Philadelphia: Churchill Livingstone; 2010. p. 3411–25. [ Google Scholar ] [ Ref list ]
5. Ali IKM, Zaki M, Clark CG. Use of PCR amplification of tRNA gene-linked short tandem repeats for genotyping Entamoeba histolytica.
J Clin Microbiol . 2005; 43 :5842–7. 10.1128/JCM.43.12.5842-5847.2005 [ PMC free article ] [ PubMed ] [ CrossRef ] [ Google Scholar ] [ Ref list ]
7. Ali IKM, Hossain MB, Roy S, Ayeh-Kumi PF, Petri WA, Haque R, et al.
Entamoeba moshkovskii infections in children, Bangladesh.
Emerg Infect Dis . 2003; 9 :580–4. 10.3201/eid0905.020548 [ PMC free article ] [ PubMed ] [ CrossRef ] [ Google Scholar ] [ Ref list ]
10. Haque R, Faruque AS, Hahn P, Lyerly DM, Petri WA. Entamoeba histolytica and Entamoeba dispar infection in children in Bangladesh.
J Infect Dis . 1997; 175 :734–6. 10.1093/infdis/175.3.734 [ PubMed ] [ CrossRef ] [ Google Scholar ] [ Ref list ]
9. Haque R, Ali IK, Akther S, Petri WA Jr. Comparison of PCR, isoenzyme analysis, and antigen detection for diagnosis of Entamoeba histolytica infection.
J Clin Microbiol . 1998; 36 :449–52. [ PMC free article ] [ PubMed ] [ Google Scholar ] [ Ref list ]
8. Haque R, Kabir M, Noor Z, Rahman SMM, Mondal D, Alam F, et al.
Diagnosis of amebic liver abscess and amebic colitis by detection of Entamoeba histolytica DNA in blood, urine, and saliva by a real-time PCR assay.
J Clin Microbiol . 2010; 48 :2798–801. 10.1128/JCM.00152-10 [ PMC free article ] [ PubMed ] [ CrossRef ] [ Google Scholar ] [ Ref list ]
6. Stensvold CR, Lebbad M, Victory EL, Verweij JJ, Tannich E, Alfellani M, et al.
Increased sampling reveals novel lineages of Entamoeba : consequences of genetic diversity and host specificity for taxonomy and molecular detection.
Protist . 2011; 162 :525–41. 10.1016/j.protis.2010.11.002 [ PubMed ] [ CrossRef ] [ Google Scholar ] [ Ref list ]

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To the Editor: Diarrheal diseases have a major effect on global health, particularly the health of malnourished children ( 1 ). The enteric parasites Entamoeba histolytica and E. moshkovskii are potential causes of diarrheal disease in children ( 2 ). For the last 20 years, we have been studying Entamoeba infections in children from the urban slum of Mirpur in Dhaka, Bangladesh ( 3 ).
E. histolytica infections can be detected through fecal microscopy, culture, PCR, and antigen detection. Microscopy and culture have limited specificity because several species of Entamoeba, which vary in their pathogenic potential, have morphologically similar cysts and trophozoites ( 4 ). In 2010–2011, during analysis of feces positive for Entamoeba organisms by microscopy or culture but negative for E. histolytica , E. dispar , and E. moshkovskii by PCR, a new species was identified, which we have named Entamoeba bangladeshi nov. sp. in recognition of the support of the Bangladesh community for this research.
Feces from both diarrheal and surveillance specimens were collected from a cohort of children living in Mirpur ( 3 ). A total of 2,039 fecal samples were examined microscopically (0.9% saline smear) and/or by fecal culture for amebic trophozoites and cysts ( 4 ). One hundred forty-nine (7%) of the samples were positive by microscopy and/or culture for an Entamoeba parasite with both cysts and trophozoites that closely resembled those of E. histolytica , E. moshkovskii , and E. dispar .
DNA was extracted directly from fecal samples by using the QIAamp DNA Stool Mini Kit (QIAGEN, Hilden, Germany) according to the manufacturer’s instructions. DNA from positive fecal cultures was isolated by using the cetyl-trimethylammonium bromide extraction method ( 5 ). PCR was conducted to detect E. histolytica , E. dispar , and E. moshkovskii , all of which are morphologically indistinguishable by microscopy and are endemic to Bangladesh ( Table ) ( 7 – 10 ). An antigen detection test (TechLab Inc. Blacksburg, VA, USA) was also used to identify fecal samples positive for E. histolytica .
* Boldface indicates the probe fluorophore and quencher.
Fecal samples (129) and cultures derived from fecal material (20) were tested by PCR. Forty-four fecal samples were positive for E. histolytica , 42 for E. dispar , and 7 for E. moshkovskii . PCR results for 48 samples were negative for all 3 parasites (mixed infections account for the total being >129); 5 cultures also were negative for all 3 parasites.
ENTAGEN-F and ENTAGEN-R primers, which exhibit a broad specificity for the small subunit ribosomal RNA (SSU rRNA) gene sequences of Entamoeba , were used in PCR to amplify DNA fragments from 43 of the samples that were negative by PCR for the 3 Entamoeba species; amplification conditions were adapted from Stensvold et al. ( 6 ). The amplified DNA was separated by electrophoresis by using 2% agarose gel. Bands of the size predicted for the Entamoeba spp. SSU rRNA gene amplicon were detected in 15 samples ( Technical Appendix Table ). The PCR products were extracted by using the QIAquick Gel Extraction Kit (QIAGEN) and cloned by using the Zero Blunt TOPO Cloning Kit (Invitrogen, Carlsbad, CA, USA). The sequenced clones from 2 different isolates, 1 diarrheal and 1 surveillance specimen, were completely novel when compared with the SSU rRNA gene sequences from other organisms and did not match any previously sequenced Entamoeba species. These isolates represent a new species of Entamoeba (GenBank accession nos. {"type":"entrez-nucleotide","attrs":{"text":"JQ412861","term_id":"402484900"}} JQ412861 and {"type":"entrez-nucleotide","attrs":{"text":"JQ412862","term_id":"402484901"}} JQ412862 ), here named E. bangladeshi ( Technical Appendix )
We examined the phylogenetic relationship between E. bangladeshi and other Entamoeba parasites by using maximum-likelihood analysis as implemented in MEGA 5 ( Technical Appendix Figure, panel A ). E. bangladeshi , although distinct, clearly grouped with the clade of Entamoeba infecting humans, including E. histolytica . E. bangladeshi , however, appeared more distantly related than the noninvasive E. dispar , but closer than E. moshkovskii, to E. histolytica .
To further characterize E. bangladeshi, we established it in xenic culture, and it displayed the ability to grow at 37°C and 25°C, a characteristic shared with E. moshkovskii and E. ecuadoriensis but that distinguishes it from E. histolytica and E. dispar . Cultured trophozoites were evaluated through light and transmission electron microscopy ( Technical Appendix Figure, panel B ). By light microscopy, we detected no apparent differences between E. bangladeshi and E. histolytica . The physical resemblance between E. histolytica and E. bangladeshi is notable because direct microscopic examination of fecal samples is still used as a diagnostic tool in areas to which these species are endemic to detect E. histolytica parasites.
Our findings add to the diversity of Entamoeba species found in humans. The incidence and effect of infection in infants by the newly recognized species E. bangladeshi await future epidemiologic studies.
Sequencing outcomes for 13 samples that were negative by PCR for Entamoeba histolytica , E. dispar , and E. moshkovskii and taxonomic summary for E. Bangladeshi .
We thank B. Mann for careful reading of this manuscript.
This investigation was supported by grant R01AI043596 from the National Institute of Allergy and Infectious Diseases to W.A.P.
Suggested citation for this article : Royer TL, Gilchrist C, Kabir M, Arju T, Ralston KS, Haque R, et al. Entamoeba bangladeshi nov. sp., Bangladesh [letter]. Emerg Infect Dis [serial on the Internet]. 2012 Sep [date cited]. http://dx.doi.org/10.3201/eid1809.120122
1 These authors contributed equally to this article.
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1. März 2023 Il dott. Spögler è stato il primo Presidente della Fondazione Cassa di Risparmio di Bolzano. Nel Suo periodo di attività, tra il 1992 e il 1994, ha dato contenuto, forma ed indirizzo all'attività del nostro ente.
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