Leishmania (L.) amazonensis infection impairs reproductive and fetal parameters in female mice / La infección por Leishmania (L.) amazonensis afecta parámetros reproductivos y fetales en ratones hembras

ABSTRACT Leishmaniasis is a group of parasitic zoonotic diseases caused by intracellular protozoans belonging to the genusLeishmania. Little is known about the effects that this parasitosis may have on the reproductive parameters and pregnancy of infected humans and pets. This study aimed to evaluate the influence of chronic cutaneous leishmaniasis caused byLeishmania (Leishmania) amazonensison reproductive and fetal parameters using a female murine model. A control group of female BALB/c mice and a group infected withL. (L.) amazonensiswere mated with healthy males. Clinical parameters were monitored during the pre-mating and gestational periods. Female mice were euthanized on day 19 of gestation, when the fetuses were weighed and their length measured and embryonic resorptions and fetal death were recorded. We observed five fetal deaths and three embryonic resorptions in the infected group. Furthermore, there was a decrease in fertility in the infected group (26.32%). The weight of the offspring from infected mothers was lower than that in the control group (1.019±0.035g and 1.163±0.032g,p<0.01). Fetal length was reduced in the infected group (3.71±0.05cm in the control group and 3.40±0.06cm in the infected groupp<0.001). This study shows that cutaneous leishmaniasis caused byL. (L.) amazonensisimpairs reproductive and fetal parameters in mice.

RESUMEN La leishmaniasis comprende un grupo de enfermedades zoonóticas parasitarias causadas por protozoos intracelulares pertenecientes al géneroLeishmania. Poco se conoce sobre los efectos que esta parasitosis puede tener sobre los parámetros reproductivos y la gestación en humanos y en otras especies infectadas. El objetivo de este estudio fue determinar la influencia de la leishmaniasis cutánea crónica, causada porLeishmania (Leishmania) amazonensis, en parámetros reproductivos y fetales. Se apareó un grupo control y un grupo infectado de ratones hembra BALB/c (previamente inoculado conL. (L.) amazonensis) con machos sanos. Se analizaron parámetros clínicos durante los períodos pregestacional y gestacional. Las hembras fueron eutanasiadas en el día 19 de gestación, momento en el cual se pesaron y midieron los fetos y se registraron las reabsorciones embrionarias y las muertes fetales. Se observaron 5 muertes fetales y 3 reabsorciones embrionarias en el grupo infectado. Además, hubo una disminución en la fertilidad de este último grupo (26,32%). Por otra parte, el peso de la descendencia de madres infectadas fue menor que el del grupo control (1,019±0,035g y 1,163±0,032g, respectivamente,p<0,01). Por último, la longitud fetal se redujo en el grupo infectado (3,71±0,05cm en el grupo control y 3,40±0,06cm en el grupo infectado,p<0,001). Este estudio muestra que la leishmaniasis cutánea causada porL. (L.) amazonensisafecta los parámetros reproductivos y fetales en ratones.

Leishmania (L.) amazonensis infection impairs reproductive and fetal parameters in female mice.

Leishmaniasis is a group of parasitic zoonotic diseases caused by intracellular protozoans belonging to the genus Leishmania. Little is known about the effects that this parasitosis may have on the reproductive parameters and pregnancy of infected humans and pets. This study aimed to evaluate the influence of chronic cutaneous leishmaniasis caused by Leishmania (Leishmania) amazonensis on reproductive and fetal parameters using a female murine model. A control group of female BALB/c mice and a group infected with L. (L.) amazonensis were mated with healthy males. Clinical parameters were monitored during the pre-mating and gestational periods. Female mice were euthanized on day 19 of gestation, when the fetuses were weighed and their length measured and embryonic resorptions and fetal death were recorded. We observed five fetal deaths and three embryonic resorptions in the infected group. Furthermore, there was a decrease in fertility in the infected group (26.32%). The weight of the offspring from infected mothers was lower than that in the control group (1.019±0.035g and 1.163±0.032g, p<0.01). Fetal length was reduced in the infected group (3.71±0.05cm in the control group and 3.40±0.06cm in the infected group p<0.001). This study shows that cutaneous leishmaniasis caused by L. (L.) amazonensis impairs reproductive and fetal parameters in mice.

Evaluation of different total Leishmania amazonensis antigens for the development of a first-generation vaccine formulated with a Toll-like receptor-3 agonist to prevent cutaneous leishmaniasis.

BACKGROUND Unfortunately, no any vaccine against leishmaniasis has been developed for human use. Therefore, a vaccine based on total Leishmania antigens could be a good and economic approach; and there are different methodologies to obtain these antigens. However, it is unknown whether the method to obtain the antigens affects the integrity and immune response caused by them. OBJECTIVES to compare the protein profile and immune response generated by total L. amazonensis antigens (TLA) produced by different methods, as well as to analyse the immune response and protection by a first-generation vaccine formulated with sonicated TLA (sTLA) and polyinosinic:polycytidylic acid [Poly (I:C)]. METHODS TLA were obtained by four different methodologies and their integrity and immune response were evaluated. Finally, sTLA was formulated with Poly (I:C) and their protective immune response was measured. FINDINGS sTLA presented a conserved protein profile and induced a strong immune response. In addition, Poly (I:C) improved the immune response generated by sTLA. Finally, sTLA + Poly (I:C) formulation provided partial protection against L. amazonensis infection. MAIN CONCLUSIONS The protein profile and immune response depend on the methodology used to obtain the antigens. Also, the formulation sTLA + Poly (I:C) provides partial protection against cutaneous leishmaniasis in mice.

Evaluation of different total Leishmania amazonensis antigens for the development of a first-generation vaccine formulated with a Toll-like receptor-3 agonist to prevent cutaneous leishmaniasis

BACKGROUND Unfortunately, no any vaccine against leishmaniasis has been developed for human use. Therefore, a vaccine based on total Leishmania antigens could be a good and economic approach; and there are different methodologies to obtain these antigens. However, it is unknown whether the method to obtain the antigens affects the integrity and immune response caused by them. OBJECTIVES to compare the protein profile and immune response generated by total L. amazonensis antigens (TLA) produced by different methods, as well as to analyse the immune response and protection by a first-generation vaccine formulated with sonicated TLA (sTLA) and polyinosinic:polycytidylic acid [Poly (I:C)]. METHODS TLA were obtained by four different methodologies and their integrity and immune response were evaluated. Finally, sTLA was formulated with Poly (I:C) and their protective immune response was measured. FINDINGS sTLA presented a conserved protein profile and induced a strong immune response. In addition, Poly (I:C) improved the immune response generated by sTLA. Finally, sTLA + Poly (I:C) formulation provided partial protection against L. amazonensis infection. MAIN CONCLUSIONS The protein profile and immune response depend on the methodology used to obtain the antigens. Also, the formulation sTLA + Poly (I:C) provides partial protection against cutaneous leishmaniasis in mice.

Detection of Trichinella britovi in pork sausage suspected to be implicated in a human outbreak in Mendoza, Argentina.

Of the three Trichinella species described in South America, T. spiralis, T. pseudospiralis and T. patagoniensis, only the former has been implicated in human infections from consumption of pork-derived products. During a presumed trichinellosis outbreak in 2012 in Mendoza, Argentina, we evaluated the serological responses of three patients who had eaten the incriminated food and had signs and symptoms compatible with trichinellosis, using ELISA. We also analyzed potentially contaminated pork sausage by artificial digestion technique and recovered Trichinella muscle larvae, which were identified to the species level using a PCR multiplex assay and by sequencing a region of the mitochondrial gene coding cytochrome oxidase subunit I. No antibodies were detected in the sera of the patients, probably because the samples were collected during the immunological window period. According to molecular identification, all larvae from the sausage corresponded to T. britovi. Trichinella britovi is reported here for the first time in the American Continent, and represents the only cold-tolerant member of the genus in the Neotropics. This species was most likely introduced from Europe to South America during Spanish colonization through pigs, wild boars and/or rats.

Diagnosis of Leishmania infection in a nonendemic area of South America.

American tegumentary leishmaniasis is the generic name for a variety of cutaneous and mucocutaneous presentations of parasitosis caused by several species of the genus Leishmania. This is a widespread infection in the American continent, from the South of the United States to the North of Argentina. We herein describe the management of a patient with American tegumentary leishmaniasis in Mendoza, Argentina, a nonendemic area of South America, whose diagnosis and treatment were significantly delayed, because the patient did not report a recent history of travel to any known endemic areas. This case stresses the need for training health-care professionals in the diagnosis and treatment of not only endemic parasitosis within their work zones but also nonendemic parasitosis.

Immunization with antigenic extracts of Leishmania associated with Montanide ISA 763 adjuvant induces partial protection in BALB/c mice against Leishmania (Leishmania) amazonensis infection.

BACKGROUND/PURPOSE: A proper adjuvant has a relevant role in vaccine formulations to generate an effective immune response. In this study, total Leishmania antigen (TLA) formulated with Montanide ISA 763 or R848 as adjuvants were evaluated as a first generation Leishmania vaccine in a murine model. METHODS: Immunization protocols were tested in BALB/c mice with a subcutaneous prime/boost regimen with an interval of 3 weeks. Mice immunized with unadjuvanted TLA and phosphate-buffered saline (PBS) served as control groups. On Day 21 and Day 36 of the protocol, we evaluated the humoral immune response induced by each formulation. Fifteen days after the boost, the immunized mice were challenged with 1 × 10(5) promastigotes of Leishmania (Leishmania) amazonensis in the right footpad (RFP). The progress of the infection was followed for 10 weeks; at the end of this period, histopathological studies were performed in the RFP. RESULTS: Vaccines formulated with Montanide ISA 763 generated an increase in the production of immunoglobulin G (IgG; p < 0.05) compared with the control group. There were no statistically significant differences in IgG1 production between the study groups. However, immunization with TLA-Montanide ISA 763 resulted in an increase in IgG2a compared to the unadjuvanted control (p < 0.001). Also noteworthy was the fact that a significant reduction in swelling and histopathological damage of the RFP was recorded with the Montanide ISA 763 formulation. CONCLUSION: We conclude that the immunization of BALB/c mice with a vaccine formulated with TLA and Montanide ISA 763 generated a protective immune response against L. (L.) amazonensis, characterized by an intense production of IgG2a.

Impact of tumor necrosis factor receptor p55 deficiency in susceptibility of C57BL/6 mice to infection with Leishmania (Leishmania) amazonensis.

Tumor necrosis factor (TNF) is involved in host resistance to several intracellular pathogens. Although the critical role of TNF receptor (TNFR)p55 in Leishmania (Leishmania) major infection has been demonstrated, the impact of TNFRp55 deficiency on L. (L.) amazonensis infection has not been explored. L. (L.) amazonensis-infected TNFRp55(-/-) mice failed to resolve lesions, whereas C57BL/6 wild-type mice completely healed. The susceptibility of the TNFRp55(-/-) mice was characterized by higher lesion size and histopathological damage in comparison with the wild-type mice. A marked increased of the splenic index was observed in the TNFRp55(-/-) mice after 15 weeks infection. These results show that in the absence of TNFRp55, L. (L.) amazonensis-infected knockout mice fail to resolve lesions, whereas wild-type mice completely heal.

Difilobotriosis humana: Un caso en área no endémica de la Argentina / Human diphyllobothriosis: A case in a non-endemic area of Argentina

La difilobotriosis es una parasitosis intestinal causada por la infección de cestodos del genero Diphyllobothrium. En la Argentina, la Patagonia Andina es considerada una zona endémica para esta parasitosis. La infección por Diphyllobothrium latum no ha sido previamente notificada en la provincia de Mendoza; en este trabajo comunicamos un caso de esta parasitosis que fue confirmada por el análisis de las características morfológicas de los huevos eliminados con la materia fecal de un paciente infectado. Se destaca la necesidad de información y capacitación de los profesionales de la salud en el diagnóstico y tratamiento de parasitosis no endémicas.(AU)

Diphyllobothriosis is an intestinal parasitosis caused by cestodes infection of the genus Diphyllobothrium. In Argentina, the Andean Patagonia is considered an endemic area for this parasitosis. Diphyllobothrium latum infection has not been previously reported in the province of Mendoza, Argentina. We are now reporting then the first case. Diphyllobothriosis was confirmed by examination of morphologic characteristics of the eggs eliminated in the patients feces. These results suggest the requirement of a more specific training of health workers in the diagnosis and treatment of non endemic parasitosis. We want to emphasize the need of health workers education on diagnosis and treatment of endemic and non-endemic parasitosis.(AU)

Difilobotriosis humana: Un caso en área no endémica de la Argentina / Human diphyllobothriosis: A case in a non-endemic area of Argentina

La difilobotriosis es una parasitosis intestinal causada por la infección de cestodos del genero Diphyllobothrium. En la Argentina, la Patagonia Andina es considerada una zona endémica para esta parasitosis. La infección por Diphyllobothrium latum no ha sido previamente notificada en la provincia de Mendoza; en este trabajo comunicamos un caso de esta parasitosis que fue confirmada por el análisis de las características morfológicas de los huevos eliminados con la materia fecal de un paciente infectado. Se destaca la necesidad de información y capacitación de los profesionales de la salud en el diagnóstico y tratamiento de parasitosis no endémicas.(AU)

Diphyllobothriosis is an intestinal parasitosis caused by cestodes infection of the genus Diphyllobothrium. In Argentina, the Andean Patagonia is considered an endemic area for this parasitosis. Diphyllobothrium latum infection has not been previously reported in the province of Mendoza, Argentina. We are now reporting then the first case. Diphyllobothriosis was confirmed by examination of morphologic characteristics of the eggs eliminated in the patients feces. These results suggest the requirement of a more specific training of health workers in the diagnosis and treatment of non endemic parasitosis. We want to emphasize the need of health workers education on diagnosis and treatment of endemic and non-endemic parasitosis.(AU)

Difilobotriosis humana: Un caso en área no endémica de la Argentina / Human diphyllobothriosis: A case in a non-endemic area of Argentina

La difilobotriosis es una parasitosis intestinal causada por la infección de cestodos del genero Diphyllobothrium. En la Argentina, la Patagonia Andina es considerada una zona endémica para esta parasitosis. La infección por Diphyllobothrium latum no ha sido previamente notificada en la provincia de Mendoza; en este trabajo comunicamos un caso de esta parasitosis que fue confirmada por el análisis de las características morfológicas de los huevos eliminados con la materia fecal de un paciente infectado. Se destaca la necesidad de información y capacitación de los profesionales de la salud en el diagnóstico y tratamiento de parasitosis no endémicas.

Diphyllobothriosis is an intestinal parasitosis caused by cestodes infection of the genus Diphyllobothrium. In Argentina, the Andean Patagonia is considered an endemic area for this parasitosis. Diphyllobothrium latum infection has not been previously reported in the province of Mendoza, Argentina. We are now reporting then the first case. Diphyllobothriosis was confirmed by examination of morphologic characteristics of the eggs eliminated in the patients' feces. These results suggest the requirement of a more specific training of health workers in the diagnosis and treatment of non endemic parasitosis. We want to emphasize the need of health workers' education on diagnosis and treatment of endemic and non-endemic parasitosis.

[Human diphyllobothriosis. A case in non-endemic area of Argentina]. / Difilobotriosis humana. Un caso en área no endémica de la Argentina.

Diphyllobothriosis is an intestinal parasitosis caused by cestodes infection of the genus Diphyllobothrium. In Argentina, the Andean Patagonia is considered an endemic area for this parasitosis. Diphyllobothrium latum infection has not been previously reported in the province of Mendoza, Argentina. We are now reporting then the first case. Diphyllobothriosis was confirmed by examination of morphologic characteristics of the eggs eliminated in the patients' feces. These results suggest the requirement of a more specific training of health workers in the diagnosis and treatment of non endemic parasitosis. We want to emphasize the need of health workers' education on diagnosis and treatment of endemic and non-endemic parasitosis.

[Trichomonas vaginalis prevalence in sex workers]. / Prevalencia de Trichomonas vaginalis en trabajadores sexuales.

Trichomoniasis is a sexually transmitted infection (STI) usually associated to vaginitis, urethritis, cervicitis and pelvic inflammatory disease, which can cause infertility. The aim of this retrospective longitudinal study was to determine Trichomonas vaginalis prevalence in a population of male and female sex workers in Mendoza, Argentina. A cohort of 720 people who attended the STI Program in this province was retrospectively analyzed. From February 2007 up to June 2009, 1692 samples were processed using a reposition sampling method. The age and sex distribution of the population was: 687 women/33 men, from 15 to 65 years old. Trichomoniasis prevalence determined for this population at risk was 7.6% in women only, men did not present infection. The high prevalence shown for this risk group suggests a great vulnerability of this population to contract other more serious STIs. In another sense, it also shows a remarkable lack in the use of mechanical barriers to prevent sexually transmitted diseases (STDs). The present research justifies the elaboration of standardized procedures guide to diagnose T. vaginalis within the framework of the Reproductive Health Program (PROSAR).

Prevalencia de Trichomonas vaginalis en trabajadores sexuales / Trichomonas vaginalis prevalence in sex workers

La tricomoniasis es una infección de transmisión sexual (ITS) que se asocia a vaginitis, uretritis, cervicitis y enfermedad inflamatoria pélvica pudiendo producir infertilidad tubárica. El objetivo de este trabajo, de tipo retrospectivo longitudinal, fue determinar la prevalencia de Trichomonas vaginalis en una población de trabajadores sexuales en Mendoza, Argentina. Se estudió en forma retrospectiva una población de 720 personas asistidas por el Programa Provincial de ITS de Mendoza. Entre febrero de 2007 y junio de 2009 se procesaron 1692 muestras con un muestreo por reposición. La distribución por sexo fue de 687 mujeres y 33 varones con edades comprendidas entre 15 y 65 años. La prevalencia de tricomoniasis determinada en esta población en riesgo fue de 7.6% para las mujeres, sin encontrarse infecciones en el grupo de los hombres. La prevalencia hallada en este grupo en riesgo muestra una población con alta vulnerabilidad para contraer otras ITS de mayor gravedad. En otro sentido, sugiere una importante falta de adhesión al uso de barreras mecánicas para prevenir infecciones de transmisión sexual. Este trabajo justifica la elaboración de una guía de procedimientos para el manejo de la infección por tricomonas en el marco del Programa de Salud Reproductiva (PROSAR).

Trichomoniasis is a sexually transmitted infection (STI) usually associated to vaginitis, urethritis, cervicitis and pelvic inflammatory disease, which can cause infertility. The aim of this retrospective longitudinal study was to determine Trichomonas vaginalis prevalence in a population of male and female sex workers in Mendoza, Argentina. A cohort of 720 people who attended the STI Program in this province was retrospectively analyzed. From February 2007 up to June 2009, 1692 samples were processed using a reposition sampling method. The age and sex distribution of the population was: 687 women/33 men, from 15 to 65 years old. Trichomoniasis prevalence determined for this population at risk was 7.6% in women only, men did not present infection. The high prevalence shown for this risk group suggests a great vulnerability of this population to contract other more serious STIs. In another sense, it also shows a remarkable lack in the use of mechanical barriers to prevent sexually transmitted diseases (STDs). The present research justifies the elaboration of standardized procedures guide to diagnose T. vaginalis within the framework of the Reproductive Health Program (PROSAR).