Abstract
Of the three species of Entamoeba found in swine, namely E. histolytica, E. polecki and E. suis, E. histolytica can also infect humans, producing colitis or abscesses of liver and leading to death. However, the pathogenicity of other species of Entamoeba has not been fully characterized. Here, we conducted histopathology and molecular surveys on a pig farm where piglets had blackish feces or muddy diarrhea. Histopathological examination of two piglets showed necrosis of the mucous surface at the ileum, cecum, or colon, infiltration of neutrophils, and formation of ulcers. Based on morphological characteristics, E. polecki and E. suis trophozoites were mainly detected at lamina propria and surface of the lesion, respectively, and Lawsonia intracellularis, a bacterial pathogen, was also detected. Molecular analysis using the small subunit ribosomal RNA gene on other piglets and a sow revealed infection with both E. polecki and E. suis. These findings corroborate our previous reports that the two Entamoeba spp. are pathogenic in pigs as aggravations of symptoms with L. intracellularis. This is the first report about mixed infection with E. polecki and E. suis.
Conflict of interest
The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
Acknowledgements
We thank Mr. M. Kobayashi and Ms. M. Shimada (National Institute of Animal Health, Japan) for assistance with histopathological observations, and we thank Dr. N. Abe (Osaka City Institute of Public Health and Environmental Sciences, Japan) for advice on molecular assays.
References
Bradford C.M., Denver M.C., Cranfield M.R. 2008. Development of a polymerase chain reaction test for Entamoeba invadens. Journal of Zoo and Wildlife Medicine, 39, 201–20710.1638/2007-0145.1Search in Google Scholar
Clark C.G., Kaffashian F., Tawari B., Windsor J.J., Twigg-Flesner A., Davies-Morel M.C., Blessmann J., Ebert F., Peschel B., Le Van A., Jackson C.J., Macfarlane L., Tannich E. 2006. New insights into the phylogeny of Entamoeba species provided by analysis of four new small-subunit rRNA genes. International Journal of Systematic and Evolutionary Microbiology, 56, 2235–223910.1099/ijs.0.64208-0Search in Google Scholar
Desowitz R.S., Barnish G. 1986. Entamoeba polecki and other intestinal protozoa in Papua New Guinea Highland children. Annals of Tropical Medicine and Parasitology, 80, 399–40210.1080/00034983.1986.11812040Search in Google Scholar
Evangelopoulos A., Spanakos G., Patsoula E., Vakalis N., Legakis N. 2000. A nested, multiplex, PCR assay for the simultaneous detection and differentiation of Entamoeba histolytica and Entamoeba dispar in faeces. Annals of Tropical Medicine and Parasitology, 94, 233–24010.1080/00034983.2000.11813534Search in Google Scholar
He G.Z., Feng Y., Deng S.X. 2012. Evaluation of the intestinal microbial diversity in miniature pig after orally infected with Entamoeba histolytica. Parasitology Research, 111, 939–94110.1007/s00436-012-2841-xSearch in Google Scholar
Jones G.F., Ward G.E., Murtaugh M.P., Lin G., Gebhart C.J. 1993. Enhanced detection of intracellular organism of swine proliferative enteritis, ileal symbiont intracellularis, in feces by polymerase chain reaction. Journal of Clinical Microbiology, 31, 2611–261510.1128/jcm.31.10.2611-2615.1993Search in Google Scholar
Kawashima K., Tsunemitsu H., Horino R., Katsuda K., Onodera T., Shoji T., Kubo M., Haritani M., Murakami Y. 2003. Effects of dexamethasone on the pathogenesis of porcine circovirus type 2 infection in piglets. Journal of Comparative Pathology, 129, 294–30210.1016/S0021-9975(03)00047-1Search in Google Scholar
Kawashima K., Yamada S., Kobayashi H., Narita M. 1996. Detection of porcine reproductive and respiratory syndrome virus and Mycoplasma hyorhinis antigens in pulmonary lesions of pigs suffering from respiratory distress. Journal of Comparative Pathology, 114, 315–32310.1016/S0021-9975(96)80053-3Search in Google Scholar
La T., Phillips N.D., Hampson D.J. 2003. Development of a duplex PCR assay for detection of Brachyspira hyodysenteriae and Brachyspira pilosicoli in pig feces. Journal of Clinical Microbiology, 41, 3372–337510.1128/JCM.41.7.3372-3375.2003Search in Google Scholar PubMed PubMed Central
León-Avila G., Tovar J. 2004. Mitosomes of Entamoeba histolytica are abundant mitochondrion-related remnant organelles that lack a detectable organellar genome. Microbiology, 150, 1245–125010.1099/mic.0.26923-0Search in Google Scholar PubMed
Matsubayashi M., Suzuta F., Terayama Y., Shimojo K., Yui T., Haritani M., Shibahara T. 2014. Ultrastructural characteristics and molecular identification of Entamoeba suis isolated from pigs with hemorrhagic colitis: implications for pathogenicity. Parasitology Research, 113, 3023–3028. 10.1007/s00436-014-3965-ySearch in Google Scholar PubMed
Matsubayashi M., Kanamori K., Sadahiro M., Tokoro M., Abe N., Haritani M., Shibahara T. 2015a. First molecular identification of Entamoeba polecki in a piglet in Japan and implications for aggravation of ileitis by coinfection with Lawsonia intracellularis. Parasitology Research, 114, 3069–3073. 10.1007/s00436-015-4509-9Search in Google Scholar PubMed
Matsubayashi M., Murakoshi N., Komatsu T., Tokoro M., Haritani M., Shibahara T. 2015b. First molecular identification of Entamoeba polecki subtype 3 (Entamoeba struthionis) from a pig in Japan and characterization of its pathogenic role in ulcerative colitis. Infection, Genetics and Evolution, 36, 8–14. 10.1016/j.meegid.2015.08.030Search in Google Scholar PubMed
Noble G.A., Noble E.R. 1952. Entamoebae in farm mammals. Journal of Parasitology, 38, 571–59510.2307/3273985Search in Google Scholar
Prophet E.B., Mills B., Arrington J.B., Sobin L.H. (Eds.). 1992. Laboratory Methods in Histotechnology. American Registry of Pathology, Washington DC, pp. 132–214Search in Google Scholar
Schuster F.L., Visvesvara G.S. 2004. Amebae and ciliated protozoa as causal agents of waterborne zoonotic disease. Veterinary Parasitology, 126, 91–12010.1016/j.vetpar.2004.09.019Search in Google Scholar PubMed
Shibahara T., Wada Y., Sueyoshi M., Ohya T., Ishikawa Y., Kadota K. 2000. Bovine intestinal spirochaetosis with dysentery. Veterinary Record, 146, 585–58610.1136/vr.146.20.585Search in Google Scholar PubMed
Shimizu C., Shibahara T., Takai S., Kasuya K., Chikuba T., Murakoshi N., Kobayashi H., Kubo M. 2010. Lawsonia intracellularis and virulent Rhodococcus equi infection in a thoroughbred colt. Journal of Comparative Pathology, 143:303–8. 10.1016/j.jcpa.2010.03.005Search in Google Scholar PubMed
Skappak C., Akierman S., Belga S., Novak K., Chadee K., Urbanski S.J., Church D., Beck P.L. 2014. Invasive amoebiasis: a review of Entamoeba infections highlighted with case reports. Canadian Journal of Gastroenterology & Hepatology, 28, 355–35910.1155/2014/745130Search in Google Scholar PubMed PubMed Central
Solaymani-Mohammadi S., Rezaian M., Hooshyar H., Mowlavi G.R., Babaei Z., Anwar M.A. 2004. Intestinal protozoa in wild boars (Sus scrofa) in western Iran. Journal of Wildlife Diseases, 40, 801–80310.7589/0090-3558-40.4.801Search in Google Scholar PubMed
Stensvold C.R., Lebbad M., Victory E.L., Verweij J.J., Tannich E., Alfellani M., Legarraga P., Clark C.G. 2011. Increased sampling reveals novel lineages of Entamoeba: consequences of genetic diversity and host specificity for taxonomy and molecular detection. Protist, 162, 525–541. 10.1016/j.protis.2010.11.002Search in Google Scholar PubMed
Tachibana H., Kobayashi S., Takekoshi M., Ihara S. 1991. Distinguishing pathogenic isolates of Entamoeba histolytica by polymerase chain reaction. Journal of Infectious Diseases, 164, 825–82610.1093/infdis/164.4.825Search in Google Scholar PubMed
Tovar J., Fischer A., Clark C.G. 1999. The mitosome, a novel organelle related to mitochondria in the amitochondrial parasite Entamoeba histolytica. Molecular Microbiology, 32, 1013–102110.1046/j.1365-2958.1999.01414.xSearch in Google Scholar PubMed
Verweij J.J., Laeijendecker D., Brienen E.A., van Lieshout L., Polderman A.M. 2003. Detection and identification of Entamoeba species in stool samples by a reverse line hybridization assay. Journal of Clinical Microbiology, 41, 5041–504510.1128/JCM.41.11.5041-5045.2003Search in Google Scholar PubMed PubMed Central
Verweij J.J., Polderman A.M., Clark C.G. 2001. Genetic variation among human isolates of uninucleated cyst-producing Entamoeba species. Journal of Clinical Microbiology, 39, 1644–164610.1128/JCM.39.4.1644-1646.2001Search in Google Scholar PubMed PubMed Central
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