Name: sexual transmission of american trypanosomes from males and females to naive mates
Uploaded: 2019-01-27T13:00:00
Description: Watch this Scientific Journal Video about Sexual Transmission of American Trypanosomes from Males and Females to Naive Mates at JoVE.com

We acknowledge the laboratory facilities and the critical comments of Izabela Dourado, Carla Araujo, and Clever Gomes and the technical assistance of Bruno Dalago and Rafael Andrade. We are indebted to the Foundation for the Advancement of Science (FAPDF), The National Research Council, Ministry of Science and Technology (CNPq/MCT), and The Agency for Training Human Resources, Ministry of Education (CAPES/ME), Brazil, for supporting these investigations.

The Human and the Animal Research Committees of the Faculty of Medicine of the University of Brasilia approved all the procedures with human subjects and laboratory animals, respectively, in research protocols 2500.167567 and 10411/2011. The Ethics Committee of the Public Foundation Hospital Gaspar Vianna (protocol n&#186; 054/2009 and CONEP 11163/2009) approved the free consent forms for the field study, with extension to the Ministry of Health National Commission on Human Research (CONEP 2585/04). The protocol was adju...

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Herein, we discuss a family-based research protocol that answered the question of whether human Chagas disease stems from sexually transmitted intraspecies T. cruzi infections. Early studies could not provide evidence of the sexual transmission of T. cruzi infections, probably because the available data and information on Chagas disease were obtained separately from the individual3,4,5,<sup class="xr...

The protozoan parasite Trypanosoma cruzi belonging to the family Trypanosomatidae undergoes trypomastigote and amastigote life cycle stages in mammalian hosts and exists as epimastigotes in the insect-vector&#39;s (Reduviid: Triatominae) gut and in axenic culture. In recent decades, several studies have shown the presence of Chagas disease in countries on four continents considered triatomine bug free1,2,3,4,5,6,7,8,9,10,11,12,13; the dispersion of American trypanosomes was initially attributed to Latin American immigrants to the Northern Hemisphere, but the possibility that some are autochthonous cases of Chagas disease can no longer be denied3,4,5,6,7,8,9,10,11,12,13,14. The only recognizable endogenous source of T. cruzi transmission has been ascribed to the chagasic mother&#39;s transfer of the parasite to the offspring in approximately 10% of pregnancies15; the male partner&#39;s contribution to in utero&#160;infections through semen ejaculates has remained unrecognized.
Over one century ago, investigators16,17 observed intracellular T. cruzi amastigotes in the theca cells of the ovary and in the germ line cells of the testicles of acute cases of Chagas disease. The nests and clumps of T. cruzi trypomastigotes and amastigotes in theca cells of the ovary, in goniablasts and in the lumen of seminiferous tubules (Figure 1) of fatal acute Chagas disease cases develop immune privilege in the organs of reproduction in the absence of inflammatory infiltrates18,19. In recent decades, a few experimental studies have shown nests of the round amastigote forms of T. cruzi in the seminiferous tubule, epididymis, and vas deferens as well as in the uterus, tubes and ovary theca cells of acutely infected mice1,20,21,22. Furthermore, in the course of family studies to document the transfer of protozoan&#160;mitochondrial DNA from parental Chagas patients to their descendants, T. cruzi nuclear DNA (nDNA) was verified in human haploid germ line cells23, and parasite life cycle stages were observed in the ejaculates of chagasic mice24. These findings are in agreement with reports on the immune tolerance attained by the progeny of T. cruzi-infected hosts in the absence of the specific antibody1,25,26. Additionally, epidemiological reports that suggested the spread of endemic Chagas disease to the other continents3,4,5,6,7,8,9,10,11,12,13 are now supported by experimental studies showing that Chagas disease can be transmitted sexually1. The present investigation presents an epidemiologic family study protocol and shows that T. cruzi infection propagates by sexual intercourse.

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		<tr height="20">
			<td height="20" style="height:20px;width:336px;">BCIP and NBT redox system</td>
			<td style="width:183px;">Sigma-Aldrich</td>
			<td style="width:102px;">681 451 001</td>
			<td style="width:65px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:336px;">Blood DNA Purification columns</td>
			<td style="width:183px;">Amersham Biosciences</td>
			<td style="width:102px;">27-9603-01</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:336px;">d-ATP, [&#945;-32P], 250 &#181;Ci.</td>
			<td style="width:183px;">&#160; Perkin Elmer&#160;&#160;</td>
			<td style="width:102px;">BLU012H</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:336px;">DNA, Solution Salt Fish Sperm</td>
			<td style="width:183px;">AMRESCO</td>
			<td style="width:102px;">064-10G</td>
			<td></td>
		</tr>
		<tr height="33">
			<td height="33" style="height:34px;width:336px;">dNTP Set, 100 mM Solutions</td>
			<td style="width:183px;">GE Healthcare</td>
			<td style="width:102px;">28-4065-51</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:336px;">Eco&#160;RI</td>
			<td style="width:183px;">Invitrogen</td>
			<td style="width:102px;">15202-021</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:336px;">Goat anti-human IgG- alkaline phosphatase conjugated</td>
			<td style="width:183px;">Southern Biotech</td>
			<td style="width:102px;">&#160;&#160;&#160;&#160;&#160;&#160; 2040-04</td>
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			<td height="20" style="height:20px;width:336px;">Micro imaging software cell Sens software</td>
			<td style="width:183px;">Olympus, Japan</td>
			<td style="width:102px;"></td>
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			<td height="20" style="height:20px;width:336px;">Plasmid Prep Mini Spin Kit</td>
			<td style="width:183px;">GE Healthcare</td>
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			<td height="20" style="height:20px;width:336px;">Plate reader&#160;</td>
			<td style="width:183px;">Bio-Tek GmBH</td>
			<td style="width:102px;">2015</td>
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		<tr height="20">
			<td height="20" style="height:20px;width:336px;">Rabbit anti-chicken IgG-alkaline phosphatase conjugated</td>
			<td style="width:183px;">Sigma-Aldrich</td>
			<td style="width:102px;">A9171</td>
			<td></td>
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		<tr height="20">
			<td height="20" style="height:20px;width:336px;">Rabbit anti-chicken IgG-FITC conjugated</td>
			<td style="width:183px;">Sigma-Aldrich</td>
			<td style="width:102px;">F8888</td>
			<td></td>
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		<tr height="20">
			<td height="20" style="height:20px;width:336px;">Rabbit anti-mouse IgG- alkaline phosphatase conjugated</td>
			<td style="width:183px;">Sigma Aldrich</td>
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		<tr height="20">
			<td height="20" style="height:20px;width:336px;">Rabbit anti-mouse IgG-FITC conjugated</td>
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			<td height="20" style="height:20px;width:336px;">Spin Columns for radio labeled DNA purification, Sephadex G-25, fine</td>
			<td style="width:183px;">Sigma-Aldrich</td>
			<td style="width:102px;">G25DNA-RO&#160;</td>
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			<td height="20" style="height:20px;width:336px;">Taq DNA Polymerase Recombinant</td>
			<td style="width:183px;">Invitrogen</td>
			<td style="width:102px;">11615-010</td>
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			<td height="20" style="height:20px;width:336px;">Thermal cycler system</td>
			<td style="width:183px;">Biorad, USA</td>
			<td style="width:102px;">1709703</td>
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			<td height="20" style="height:20px;width:336px;">Vector Systems</td>
			<td style="width:183px;">Promega</td>
			<td style="width:102px;">A1380</td>
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	</tbody>
</table>

sexual transmission of american trypanosomes from males and females to naive mates

American trypanosomiasis is transmitted to humans by triatomine bugs through the ingestion of contaminated food, by blood transfusions or accidently in hospitals and research laboratories. In addition, the Trypanosoma cruzi infection is transmitted congenitally from a&#160;chagasic mother to her offspring, but the male partner&#39;s contribution to in utero contamination is unknown. The findings of nests and clumps of amastigotes and of trypomastigotes in the theca cells of the ovary, in the goniablasts and in the lumen of seminiferous tubules suggest that T. cruzi infections are sexually transmitted. The research protocol herein presents the results of a family study population showing parasite nuclear DNA in the diploid blood mononuclear cells and in the haploid gametes of human subjects. Thus, three independent biological samples collected one year apart confirmed that T. cruzi infections were sexually transmitted to progeny. Interestingly, the specific T. cruzi antibody was absent in the majority of family progeny that bore immune tolerance to the parasite antigen. Immune tolerance was demonstrated in chicken refractory to T. cruzi after the first week of embryonic growth, and chicks hatched from the flagellate-inoculated eggs were unable to produce the specific antibody.&#160;Moreover, the instillation of the human semen ejaculates intraperitoneally or into the vagina of naive mice yielded T. cruzi amastigotes in the epididymis, seminiferous tubule, vas deferens and uterine tube with an absence of inflammatory reactions in the immune privileged organs of reproduction. The breeding of T. cruzi-infected male and female mice with naive mates resulted in acquisition of the infections, which were later transmitted to the progeny. Therefore, a robust education, information and communication program that involves the population and social organizations is deemed necessary to prevent Chagas disease.

This research, conducted according to the protocol, aimed to detect acute cases of Chagas disease by clinical and parasitological exams. Venous blood samples were subjected to direct microscopic examination and in vitro culture for parasite growth. Twenty-one acute cases of Chagas disease showed T. cruzi in blood. The research protocol secured the isolation of T. cruzi ECI1-to ECI21 from acute Chagas disease, and the DNA samples exhibited positive DNA footprints in the r...

Watch this Scientific Journal Video about Sexual Transmission of American Trypanosomes from Males and Females to Naive Mates at JoVE.com

Sexual Transmission of American Trypanosomes from Males and Females to Naive Mates

The Trypanosoma cruzi agent of Chagas disease produces long-lasting asymptomatic infections that abruptly develop into clinically recognized pathology. The following research protocol describes a short-run family-based epidemiological study to unravel the T. cruzi infection transmitted sexually from parent to progeny.

American trypanosomiasis is transmitted to humans by triatomine bugs through the ingestion of contaminated food, by blood transfusions or accidently in ...

Research

JoVE Journal

Immunology and Infection

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