Allele diversity of the 24-<i>loci</i> MIRU-VNTR.

<p>Allele diversity of the 24-<i>loci</i> MIRU-VNTR.</p

Molecular characterization of the 37 <i>M</i>. <i>bovis</i> isolates by spoligotyping method.

<p>Molecular characterization of the 37 <i>M</i>. <i>bovis</i> isolates by spoligotyping method.</p

Dendrogram generated by the <i>BioNumerics</i> 6.6 software (Applied Maths) based on the combination of spoligotyping and MIRU-VNTR analyses applied to the 37 <i>M</i>. <i>bovis</i> isolates, using the categorical index and unweighted pair-grouping method analysis algorithm (UPGMA).

<p>Dendrogram generated by the <i>BioNumerics</i> 6.6 software (Applied Maths) based on the combination of spoligotyping and MIRU-VNTR analyses applied to the 37 <i>M</i>. <i>bovis</i> isolates, using the categorical index and unweighted pair-grouping method analysis algorithm (UPGMA).</p

Molecular characterization of <i>M</i>. <i>bovis</i> isolates from cattle in Midwest Brazil.

<p>Molecular characterization of <i>M</i>. <i>bovis</i> isolates from cattle in Midwest Brazil.</p

Discriminatory ability comparison among the spoligotyping and 24 MIRU-VNTR methods and the combination of both in detecting genetic similarities.

<p>Discriminatory ability comparison among the spoligotyping and 24 MIRU-VNTR methods and the combination of both in detecting genetic similarities.</p

Numbers of <i>M</i>. <i>leprae</i> recovered from mice footpads after a six month infection with 10⁴ bacillus isolated from <i>Rhodnius prolixus</i> feces that received an infectious blood meal.

<p>Numbers of <i>M</i>. <i>leprae</i> recovered from mice footpads after a six month infection with 10⁴ bacillus isolated from <i>Rhodnius prolixus</i> feces that received an infectious blood meal.</p

<i>M</i>. <i>leprae</i> reaches <i>Rhodnius prolixus</i> feces 20 days after infection.

<p><i>Rhodnius prolixus</i> were allowed to feed rabbit blood containing (A-B) or not (C-D) 10⁷ PKH26-<i>M</i>. <i>leprae</i> / mL. During the next non-infected blood meal (20 days after infection), feces from groups of ten insects were collected in a sterile tube and analyzed by fluorescence microscopy (B and D). Stained <i>M</i>. <i>leprae</i> was identified only in the infected insects (A-B), allowing its counting. Images are representative of six pools of feces, where at least five fields were analyzed. Scale bar indicates 20μm.</p

The kissing bug <i>Rhodnius prolixus</i> is able to maintain <i>M</i>. <i>leprae</i> viability inside its digestive tract after infection.

<p><i>M</i>. <i>leprae</i> viability was determined by the persistence of 16Sr RNA at the different digestive compartments of artificially infected adult <i>Rhodnius prolixus</i>: anterior midgut (spheres), posterior midgut (squares) and hindgut (triangles), just after blood meal (2h) and after total blood meal digestion (20 days), infected with the pathogen. As we can see, the hindgut was the only compartment where the level of living <i>M</i>. <i>leprae</i> increases after 20 days of infection. Scatter plot showing mean and SEM of four independent experiments, each point represent five insects group. *** means p < 0.001 (2h x 20 days, for each group of sample). Non-infected controls did not present amplification of the targets.</p

Experimental Infection of <i>Rhodnius prolixus</i> (Hemiptera, Triatominae) with <i>Mycobacterium leprae</i> Indicates Potential for Leprosy Transmission

<div><p>Leprosy is a chronic dermato-neurological disease caused by infection with <i>Mycobacterium leprae</i>. In 2013 almost 200,000 new cases of leprosy were detected around the world. Since the first symptoms take from years to decades to appear, the total number of asymptomatic patients is impossible to predict. Although leprosy is one of the oldest records of human disease, the mechanisms involved with its transmission and epidemiology are still not completely understood. In the present work, we experimentally investigated the hypothesis that the mosquitoes <i>Aedes aegypti</i> and <i>Culex quinquefasciatus</i> and the hemiptera <i>Rhodnius prolixus</i> act as leprosy vectors. By means of real-time PCR quantification of <i>M</i>. <i>leprae</i> 16SrRNA, we found that <i>M</i>. <i>leprae</i> remained viable inside the digestive tract of <i>Rhodnius prolixus</i> for 20 days after oral infection. In contrast, in the gut of both mosquito species tested, we were not able to detect <i>M</i>. <i>leprae</i> RNA after a similar period of time. Inside the kissing bug <i>Rhodnius prolixus</i> digestive tract, <i>M</i>. <i>leprae</i> was initially restricted to the anterior midgut, but gradually moved towards the hindgut, in a time course reminiscent of the life cycle of <i>Trypanosoma cruzi</i>, a well-known pathogen transmitted by this insect. The maintenance of <i>M</i>. <i>leprae</i> infectivity inside the digestive tract of this kissing bug is further supported by successful mice footpad inoculation with feces collected 20 days after infection. We conclude that <i>Rhodnius prolixus</i> defecate infective <i>M</i>. <i>leprae</i>, justifying the evaluation of the presence of <i>M</i>. <i>leprae</i> among sylvatic and domestic kissing bugs in countries endemic for leprosy.</p></div

Description of clusters containing 3 or more isolates in this study and their worldwide distribution in the SITVIT2 database (interrogation made on September 25^th 2013).

<p>*Worldwide distribution is reported for regions with more than 3% of a given SITs as compared to their total number in the SITVIT2 database. The definition of macro-geographical regions and sub-regions (<a href="http://unstats.un.org/unsd/methods/m49/m49regin.htm" target="_blank">http://unstats.un.org/unsd/methods/m49/m49regin.htm</a>) is according to the United Nations; Regions: AFRI (Africa), AMER (Americas), ASIA (Asia), EURO (Europe), and OCE (Oceania), subdivided in: E (Eastern), M (Middle), C (Central), N (Northern), S (Southern), SE (South-Eastern), and W (Western). Furthermore, CARIB (Caribbean) belongs to Americas, while Oceania is subdivided in 4 sub-regions, AUST (Australasia), MEL (Melanesia), MIC (Micronesia), and POLY (Polynesia). Note that in our classification scheme, Russia has been attributed a new sub-region by itself (Northern Asia) instead of including it among rest of the Eastern Europe. It reflects its geographical localization as well as due to the similarity of specific TB genotypes circulating in Russia (a majority of Beijing genotypes) with those prevalent in Central, Eastern and South-Eastern Asia.</p><p>**The 3 letter country codes are according to <a href="http://en.wikipedia.org/wiki/ISO_3166-1_alpha-3" target="_blank">http://en.wikipedia.org/wiki/ISO_3166-1_alpha-3</a>; countrywide distribution is only shown for SITs with ≥3% of a given SITs as compared to their total number in the SITVIT2 database.</p><p>Description of clusters containing 3 or more isolates in this study and their worldwide distribution in the SITVIT2 database (interrogation made on September 25^th 2013).</p