Coinfection by Trypanosoma cruzi and a fungal pathogen increases survival of Chagasic bugs: advice against a fungal control strategy.

Triatomine bugs carry the parasitic protozoa Trypanosoma cruzi, the causal agent of Chagas disease. It is known that both the parasite and entomopathogenic fungi can decrease bug survival, but the combined effect of both pathogens is not known, which is relevant for biological control purposes. Herein, the survival of the triatomine Meccus pallidipennis (Stal, 1872) was compared when it was coinfected with the fungus Metarhizium anisopliae (Metschnikoff) and T. cruzi, and when both pathogens acted separately. The immune response of the insect was also studied, using phenoloxidase activity in the bug gut and hemolymph, to understand our survival results. Contrary to expectations, triatomine survival was higher in multiple than in single challenges, even though the immune response was lower in cases of multiple infection. We postulate that T. cruzi exerts a protective effect and/or that the insect reduced the resources allocated to defend itself against both pathogens. Based on the present results, the use of M. anisopliae as a control agent should be re-considered.

In vitro activity of steroidal dendrimers on Trypanosoma cruzi epimastigote form with PAMAM dendrons modified by "click" chemistry.

The increasing use of dendrimers shows promise for the treatment of inflammatory diseases, Chagas disease and other conditions such as cancer. In this study, the activity of 1st and 2nd generation dendrimers over T. cruzi in the epimastigote stage was tested. Dendrimers were derived from α-ethynylestradiol (EE) modified with PAMAM-type dendrons through a triazole ring. The activity of each compound was evaluated in five doses (from 1.3 to 20 µmol/mL) by flow cytometry, including benznidazole (Bz) as positive control. The findings show that an equivalent concentration of 14.8 µmol/mL of 2nd generation (G) dendrimer is 8 times more effective than Bz at 24 h, and it maintains its superiority at 48 h with an IC50 = 1.25 ±â€¯0.19 µmol/mL. A TUNEL assay showed that dendrimers induce cell death in T. cruzi epimastigotes mostly via apoptosis, unlike Bz, which induces death via necrosis in more than 50% of cells.

The cost of being a killer's accomplice: Trypanosoma cruzi impairs the fitness of kissing bugs.

Relatively little is known about the fitness effects and life history trade-offs in medically important parasites and their insect vectors. One such case is the triatomine bugs and the parasite Trypanosoma cruzi, the key actors in Chagas disease. Previous studies have revealed some costs but have not simultaneously examined traits related to development, reproduction, and survival or their possible trade-offs. In addition, these studies have not compared the effects of genetically different T. cruzi strains that differ in their weakening effects in their vertebrate hosts. We compared the body size of the bugs after infection, the number of eggs laid, hatching/non-hatching rate, hatching success, survival, and the resulting number of parasites in Meccus (Triatoma) pallidipennis bugs that were experimentally infected with two strains of T. cruzi (Chilpancingo [CH], the most debilitating in vertebrates; and Morelos [MO], the least debilitating) (both belonging to TcI group). Our results showed that infection affects size (MO < CH; MO and CH = control), number of eggs laid (MO and CH < control) hatching/non-hatching rate (MO < control < CH), hatching success (control < MO, CH = control = MO), and survival (Chilpancingo < Morelos < control). In addition, the CH strain produced more parasites than the MO strain. These results suggest that (a) infection costs depend on the parasite's origin, (b) the more debilitating effects of the CH strain are due to its increased proliferation in the host, and (c) differences in pathogenicity among T. cruzi strains can be maintained through their different effects on hosts' life history traits. Probably, the vectorial capacity mediated by a more aggressive strain could be reduced due to its costs on the triatomine, leading to a lower risk of vertebrate and invertebrate infection in natural populations.

[Enfermedad de Chagas en México].

Chagas disease, which is caused by Trypanosoma cruzi, is considered to be the most serious parasitic disease in America. It is transmitted mainly by triatominae ("kissing bugs"). Mazzoti reported the first two human cases in Mexico. The form of transmission is by parasites entering the organism in feces of the insect, by blood transfusion, from mother to child, by organ transplant and laboratory accidents. In Mexico, 1.1 million people are estimated to be infected; the incidence in 2012 was 0.70 per 1,00,000 population. In 2017, the highest incidence rates were registered in Yucatán, Oaxaca and Hidalgo. The infection causes cardiomyopathies and mega-organs of the digestive tract. Diagnosis in the acute phase is by parasitological approach and, in the chronic phase, by laboratory screening studies. In Mexico's blood banks, screening for Chagas disease is mandatory; from 2007 to 2016, seroprevalence has decreased from 0.40 to 0.32 due to the improvement of donor selection processes and the ad hoc questionnaire. The targets of the parasite are neurons and smooth and myocardial muscle cells. The association of neuronal and smooth muscle destruction defines the presentation of chagas mega-syndromes. Initial manifestations of the disease can go unnoticed; 5% show apparent signs and symptoms and 30% will progress to the chronic asymptomatic phase. Currently available treatments have effect in the acute phase. For the control of Chagas disease, the Specific Action Program for the Prevention and Control of Chagas Disease (PAE Chagas 2013-2018) is available to initiate activities aimed at eliminating transfusion and congenital transmission and controlling vector transmission. The success of medical care depends on oportune detection, early etiological treatment and coverage broadening. On the other hand, monitoring and screening of pregnant women living in risk areas and blood and organ donors universal screening will enable the elimination congenital and transfusion transmission.

La enfermedad de Chagas, causada por el Trypanosoma cruzi, está considerada como la parasitosis más grave en América. Se transmite principalmente por triatominos (chinches). El doctor Mazzoti reportó los dos primeros casos humanos en México. La forma de transmisión es por la entrada al organismo de los parásitos en heces del insecto, por transfusión sanguínea, de madre a hijo, por trasplante de órganos y por accidentes de laboratorio. En México se estima que 1.1 millones de personas están infectadas; la incidencia en 2012 fue de 0.70 por 100 000 habitantes. En 2017, las mayores tasas de incidencia se registraron en Yucatán, Oaxaca e Hidalgo. La infección ocasiona miocardiopatías y megaórganos del tracto digestivo. El diagnóstico en fase aguda es por abordaje parasitológico y en fase crónica, por estudios de tamizaje por laboratorio. En los bancos de sangre de México, el estudio de la enfermedad de Chagas es de observancia obligatoria; de 2007 a 2016, la seroprevalencia ha disminuido de 0.40 a 0.32 debido a la mejora de los procesos de selección al donante y al cuestionario ad hoc. Los blancos del parásito son las células neuronales y las de los músculos liso y miocárdico. La asociación de la destrucción neuronal y del músculo liso define la presentación de los síndromes megachagásicos. Las manifestaciones iniciales de la enfermedad pueden pasar desapercibidas; 5 % de los pacientes presenta signos y síntomas aparentes y 30 % evolucionará a la fase crónica asintomática. Los tratamientos actuales tienen efecto en la fase aguda. Para el control de la enfermedad de Chagas se dispone del Programa de Acción Específico para la Vigilancia Prevención y Control de la Enfermedad de Chagas (PAE Chagas 2013-2018), encaminado a eliminar la transmisión transfusional y congénita y a controlar la transmisión vectorial. De la detección oportuna, el tratamiento etiológico temprano y la ampliación de cobertura depende el éxito de la atención médica. Por su parte, la vigilancia y tamizaje de las mujeres embarazadas que viven en zonas de riesgo y el tamizaje universal de donadores de sangre y órganos harán posible la eliminación de la transmisión connatal y transfusional.

Chagas Disease in Mexico: Report of 14 Cases of Chagasic Cardiomyopathy in Children.

Chagas disease is a parasitic infection mainly found in Latin America; it is transmitted by a triatomine, also known as assassin bug or kissing bug. In humans, the parasite causes mostly cardiac disorders. Two-thirds of the Mexican territory are regarded as risk areas for vector transmission of Trypanosoma cruzi, the causal agent. The parasite can be found as a blood-borne trypomastigote or as an intracellular amastigote. The progression and severity of lesions could be due to frequent reinfections or to infection by highly virulent strains. A total of 3,327 individuals younger than 18 years old, living in risk areas for this disease in the rural setting of the States of Queretaro, San Luis Potosi, and Veracruz, underwent a seroepidemiological study. Among them, 37 subjects were seropositive for T. cruzi, and were studied to look for signs of cardiac pathology, which has only been reported in adults. A clinical record was prepared for all included individuals, and electrocardiography (ECG) and echocardiography (ECHO) studies were performed; 25 cases showed lesions compatible with the onset of Chagas cardiomyopathy. The other 12 patients showed either normal ECG and ECHO data or showed abnormal parameters that were not regarded as significant. Lesions found in the onset of Chagas cardiomyopathy in children are herein reported, along with 14 cases of cardiac pathology compatible with Chagas disease. Our results indicate that patients younger than 18 years can show a cardiac pathology similar to that observed in adults.

[Identification of immunodominant components of an isolate of Trypanosoma cruzi by immunoblot and its standardization for diagnostic purposes]. / Identificación de componentes inmunodominantes de un aislado de Trypanosoma cruzi por inmunoblot y su estandarización con fines diagnósticos.

INTRODUCTION: Conventional serology was used for the detection of Trypanosoma cruzi infection, with diverse sensitivity and specificity results. Due to the number of samples with doubtful results, it is necessary to develop additional confirmation tests such as the immunoblot. OBJECTIVE: The aim of this study was identify major immunogenic proteins of T. cruzi isolate and establish criteria for immunoblot positivity with diagnostic purposes. METHODS: Immunoblot initial standardization was performed, determining optimal concentrations of antigen, serum, and second antibody. Thirty-five positive and thirty negative sera were assayed to evaluate different criteria of positivity and determine which provides greater sensitivity and specificity. RESULTS: Immunoblot of T. cruzi positive sera shared a rich pattern of components with molecular weights between 10-250 kDa. Twelve components had a recognition rate higher than 50%, of which the polypeptides of 27, 32, 34, and 38 kDa were close to 100%. Of the positivity criteria evaluated, the recognition of the components of 27 and 32 kDa provided sensitivity and specificity of 100%. DISCUSSION: The Immunoblot is suitable for confirmation of infection by T. cruzi, so it is strongly recommended for confirmation and discrimination of discordant cases.

Expression of Proteins, Glycoproteins, and Transcripts in the Guts of Fasting, Fed, and Trypanosoma cruzi-Infected Triatomines: A Systematic Review.

Chagas disease is caused by the hemoflagellate protozoan Trypanosoma cruzi. The main transmission mechanism for the parasite in endemic areas is contact with the feces of an infected triatomine bug. Part of the life cycle of T. cruzi occurs in the digestive tract of triatomines, where vector and parasite engage in a close interaction at a proteomic-molecular level. This interaction triggers replication and differentiation processes in the parasite that can affect its infectivity for the vertebrate host. With the aim of compiling and analyzing information from indexed publications on transcripts, proteins, and glycoproteins in the guts of fasting, fed, and T. cruzi-infected triatomines in the period 2000-2022, a systematic review was conducted following the PRISMA guidelines. Fifty-five original research articles retrieved from PubMed and ScienceDirect were selected; forty-four papers reported 1-26,946 transcripts, and twenty-one studies described 1-2603 peptides/proteins.

Immunopathological Mechanisms Underlying Cardiac Damage in Chagas Disease.

In Chagas disease, the mechanisms involved in cardiac damage are an active field of study. The factors underlying the evolution of lesions following infection by Trypanosoma cruzi and, in some cases, the persistence of its antigens and the host response, with the ensuing development of clinically observable cardiac damage, are analyzed in this review.

Main Cardiac Histopathologic Alterations in the Acute Phase of Trypanosoma cruzi Infection in a Murine Model.

Symptoms in the acute phase of Chagas disease are usually mild and nonspecific. However, after several years, severe complications like dilated heart failure and even death may arise in the chronic phase. Due to the lack of specific symptoms in the acute phase, the aim of this work was to describe and analyze the cardiac histopathology during this phase in a CD1 mouse model by assessing parasitism, fibrotic damage, and the presence and composition of a cellular infiltrate, to determine its involvement in the pathogenesis of lesions in the cardiac tissue. Our results indicate that the acute phase lasts about 62 days post-infection (dpi). A significant increase in parasitemia was observed since 15 dpi, reaching a maximum at 33 dpi (4.1 × 106). The presence of amastigote nests was observed at 15-62 dpi, with a maximum count of 27 nests at 35 dpi. An infiltrate consisting primarily of macrophages and neutrophils was found in the cardiac tissue within the first 30 days, but the abundance of lymphocytes showed an 8 ≥ fold increase at 40-62 dpi. Unifocal interstitial fibrosis was identified after 9 dpi, which subsequently showed a 16 ≥ fold increase at 40-60 dpi, along with a 50% mortality rate in the model under study. The increased area of fibrotic lesions revealed progression in the extent of fibrosis, mainly at 50-62 dpi. The presence of perivasculitis and thrombus circulation disorders was seen in the last days (62 dpi); finally, cases of myocytolysis were observed at 50 and 62 dpi. These histopathological alterations, combined with collagen deposition, seem to lead to the development of interstitial fibrosis and damage to the cardiac tissue during the acute phase of infection. This study provides a more complete understanding of the patterns of histopathological abnormalities involved in the acute phase, which could help the development of new therapies to aid the preclinical tests of drugs for their application in Chagas disease.

Parasitemia and Differential Tissue Tropism in Mice Infected with Trypanosoma cruzi Isolates Obtained from Meccus phyllosoma in the State of Oaxaca, Mexico.

Trypanosoma cruzi is a parasite transmitted by the feces of triatomines. Many triatomine species are found in Mexico, and various T. cruzi variants have been isolated from these species, each showing very different virulence and cell tropism. The isolates were obtained from Meccus phyllosoma specimens in three localities in the state of Oaxaca, Mexico: Tehuantitla, Vixhana, and Guichivere. The virulence of each isolate was assessed by quantifying parasitemia, survival, and histopathologic findings. The lineage of each isolate was identified using the mini-exon gene. The expression of the tssa gene during infection was detected in the heart, esophagus, gastrocnemius, and brain. Our results show that the maximum post-infection parasitemia was higher for the Tehuantitla isolate. On genotyping, all isolates were identified as T. cruzi I. The amastigotes in the heart and gastrocnemius were verified for all isolates, but in the brain only for Tehuantitla and Vixhana. The tssa expression allowed us to detect T. cruzi isolates, for Tehuantitla, predominantly in the heart. For Vixhana, a higher tssa expression was detected in gastrocnemius, and for Guichivere, it was higher in the esophagus. Results show that virulence, tropism, and tssa expression can vary, even when the isolates are derived from the same vector species, in the same region, and at similar altitudes.

[Chagas disease in Mexico. Report of two acute cases]. / Presentación de dos casos de enfermedad de Chagas aguda en México.

Two cases of acute Chagas disease in schoolchildren of 6 and 13 years of age, both with the clinical features of Romaña's sign, regional lymphadenopathy, and fever, have a history of coexistence and the bite of the transmitter, and live in housing constructed with material considered at risk for infestation by the vector; i.e. roof and walls with palma/zacate (palm tree, grass leaves), dirt floor, and inadequate illumination and ventilation. The parasitological diagnosis of both cases was established by identification of trypomastigotes of Trypanosoma cruzi in blood smears and the parasite was isolated in one of them. Benznidazole treatment was administered according to the guidelines of the WHO/PAHO for this disease. The presence of acute Chagas disease in rural areas confirms the active transmission of the parasite, so surveillance and epidemiological controls should be applied. The importance of these cases is that T. cruzi infection is symptomatic in 5% of cases, which implies that a high percentage of cases of infection appear asymptomatic and are not being diagnosed in our country.

Higher temperatures reduce the number of Trypanosoma cruzi parasites in the vector Triatoma pallidipennis.

BACKGROUND: Relatively little is known about how pathogens transmitted by vector insects are affected by changing temperatures analogous to those occurring in the present global warming scenario. One expectation is that, like their ectothermic vectors, an increase in temperature could reduce their fitness. Here, we have investigated the effect of high temperatures on the abundance of Trypanosoma cruzi parasites during infection in the vector Triatoma pallidipennis. METHODS: We exposed T. pallidipennis nymphs to two strains (Morelos and Chilpancingo) of T. cruzi. Once infected, the fifth-instar bugs were distributed among three different temperature groups, i.e. 20, 30, and 34 °C, and the resulting parasites were counted when the bugs reached adulthood. RESULTS: The number of parasites increased linearly with time at 20 °C and, to a lesser extent, at 30 °C, especially in the Chilpancingo compared to the Morelos strain. Conversely, at 34 °C, the number of parasites of both strains decreased significantly compared to the other two temperatures. CONCLUSIONS: These results suggest negative effects on the abundance of T. cruzi in T. pallidipennis at high temperatures. This is the first evidence of the effect of high temperatures on a pathogenic agent transmitted by an insect vector in the context of global warming. Further tests should be done to determine whether this pattern occurs with other triatomine species and T. cruzi strains.

Response to Infection by Trypanosoma cruzi in a Murine Model.

Cardiopathy is a common, irreversible manifestation of the chronic phase of Chagas disease; however, there is controversy as to how the causes for progression from the acute to the chronic phase are defined. In this work, the presence of the parasite is correlated with the occurrence of cell infiltration and fibrosis in cardiac tissues, as well as IgG detection and disease progression in a murine model. Fifty CD1 mice were infected intraperitoneally with Trypanosoma cruzi, while 30 control were administered with saline solution. Parasitemia levels were determined, and IgG titers were quantified by ELISA. At different times, randomly selected mice were euthanized, and the heart was recovered. Cardiac tissue slides were stained with HE and Masson trichrome stain. A significant increase in parasitemia levels was observed after 15 days post-infection (dpi), with a maximum of 4.1 × 106 parasites on 33 dpi, ending on 43 dpi; amastigote nests were observed on 15-62 dpi. Histological analysis revealed lymphocytic infiltration and fibrotic lesions from 8 dpi until the end of the study, on 100 dpi. The presence of plasma cells in the myocardium observed on 40-60 dpi, accompanied by seropositivity to ELISA on 40-100 dpi, was regarded as the hallmark of the transition phase. Meanwhile, the chronic phase, characterized by the absence of amastigotes, presence of cell infiltration, fibrotic lesions, and seropositivity, started on 62 dpi. A strong correlation between parasitemia and the presence of amastigote nests was found (r 2 = 0.930), while correlation between the presence of fibrosis and of amastigote nests was weak (r 2 = 0.306), and that between fibrosis and lymphocyte infiltration on 100 dpi was strong (r 2 = 0.899). The murine model is suitable to study Chagas disease, since it can reproduce the chronic and acute phases of the human disease. The acute phase was determined to occur on 1-60 dpi, while the chronic phase starts on 62 dpi, and fibrotic damage is a consequence of the continuous inflammatory infiltration; on the other hand, fibrosis was determined to start on the acute phase, being more apparent in the chronic phase, when Chagas disease-related cardiopathy is induced.

Occurrence of hybrids and laboratory evidence of fertility among three species of the Phyllosoma complex (Hemiptera: Reduviidae) in Mexico.

In seven studied communities of Western Mexico, triatomine specimens were sympatrically collected, some with atypical morphological characteristics in contrast to pure specimens, which were presumed to be hybrids. More than 200 specimens of Meccus pallidipennis and Meccus longipennis with brown-yellow markings on dorsal connexival segments were collected in Ahuacapán and Quitupan. In La Mesa, more than 60 specimens similar to Meccus picturatus in most morphological characteristics (including size) were collected, although they presented a largely yellowish corium like M. pallidipennis. Interfertility was proven between all of the studied wild hybrid specimens, as well as between all the experimental laboratory hybrids. Two different phenotypes (M. picturatus and M. longipennis) were obtained from crosses between M. picturatus x M. picturatus and M. longipennis x M. longipennis from the three studied localities in state of Nayarit as from La Mesita. Results support the hypothesis that the subspecific ranking of those triatomines may, therefore, be more appropriate because reproductive isolation has not been developed and complete interbreeding was recorded.

[Evaluation of the function and ventricular synchrony in patients with latency stage of Chagas' disease]. / Evaluación de la función y sincronía de la contracción ventricular en pacientes con enfermedad de Chagas en estadio de latencia.

OBJECTIVE: To compare the left ventricular function and the ventricular synchrony in patients with Chagas disease in latency stage respect to a control group. METHODS: We analyze a prospective, comparative, transversal and non randomized study of the left ventricular function (LVF) and the ventricular contraction synchronicity (VCS) in 36 subjects with positive serology for Chagas disease (18 males and 18 females), with mean of 15 +/- 5-years-old. The findings were compared with respect to 23 control volunteers (11 males and 12 females) with mean of 28 +/- 5-years-old. LVF and VCS were evaluated using equilibrium radionuclide angiography images (ERNA). The comparison of both Chagas and control populations was carried out by t Student test for independent samples, considering a statistically significant value of p < 0.05. RESULTS: The parameters of the ventricular function and the ventricular synchronicity in subjects with positive serology for Chagas disease were not statistically different with respect to the parameters of the control group. However, although they have a homogeneous contraction, the mean time of contraction for the right and the left ventricle is statistically smaller with respect to the control group. CONCLUSIONS: In clinically incipient stages of Chagas disease we do not found abnormalities in the ventricular function and the ventricular synchronicity. It's necessary to consider the follow up of the studied populations using indices for the identification of abnormalities of the autonomic nervous system.

Activity of the prophenoloxidase system and survival of triatomines infected with different Trypanosoma cruzi strains under different temperatures: understanding Chagas disease in the face of climate change.

BACKGROUND: Little is known about how human disease vectors will modify their life history patterns and survival capacity as a result of climate change. One case is that of Chagas disease, which has triatomine bugs and Trypanosoma cruzi as vectors and parasite, respectively. This work aimed to determine: (i) the activity of the prophenoloxidase system (prophenoloxidase and phenoloxidase activity, two indicators of immune ability) in three intestine regions (anterior midgut, posterior midgutand rectum) of the triatomine bug Meccus pallidipennis under three temperature conditions (20 °C, 30 °C and 34 °C) against two T. cruzi strains [ITRI/MX/14/CHIL (Chilpancingo) and ITRI/MX/12/MOR (Morelos)], and (ii) whether vector survival varies under these three temperatures after infection by these T. cruzi strains. RESULTS: Our results indicate that prophenoloxidase activity was lower at higher temperatures, that the level of prophenoloxidase activity elicited by each strain was different (higher in Chilpancingo than in Morelos strains), and that prophenoloxidase activity was more intense in the anterior midgut than in the posterior midgut or rectum. Survival rates were lower in insects maintained at higher temperatures and infected by Chilpancingo strains. CONCLUSIONS: These results indicate that climate change could lead to lower prophenoloxidase activity and survival rates in triatomines when infected with different T. cruzi strains, which could reduce the vector capacity of M. pallidipennis.

Zombie bugs? Manipulation of kissing bug behavior by the parasite Trypanosoma cruzi.

The parasite manipulation hypothesis states that the parasite modifies host's behavior thereby increasing the probability that the parasite will pass from an intermediate host to its final host. We used the kissing bugs Triatoma pallidipennis and T. longipennis and two isolates of the Trypanosoma cruzi parasite (Chilpancingo and Morelos) to test these ideas. These insects are intermediate hosts of this parasite, which is the causal agent of Chagas disease. The Chilpancingo isolate is more pathogenic than the Morelos isolate, in the bugs. We expected that infected bugs would be more active and likely at detecting human-like odors. Given the differences in pathogenicity between isolates, we expected the Chilpancingo isolate to induce these effects more strongly and lead to higher parasite number than the Morelos isolate. Finally, infected bugs would gain less mass (a mechanism thought to increase bite rate, and thus transmission) than non-infected bugs. Having determined that both isolate haplotypes belong to the Tc1a group, we found that: (a) young instars of both species were more active and likely to detect human odor when they were infected, regardless of the isolate; (b) there was no difference in parasite abundance depending on isolate; and, (c) infected bugs did not end up with less weight than uninfected bugs. These results suggest that T. cruzi can manipulate the bugs, which implies a higher risk to contract Chagas disease than previously thought.

Identification of O-Glcnacylated Proteins in Trypanosoma cruzi.

Originally an anthropozoonosis in the Americas, Chagas disease has spread from its previous borders through migration. It is caused by the protozoan Trypanosoma cruzi. Differences in disease severity have been attributed to a natural pleomorphism in T. cruzi. Several post-translational modifications (PTMs) have been studied in T. cruzi, but to date no work has focused on O-GlcNAcylation, a highly conserved monosaccharide-PTM of serine and threonine residues mainly found in nucleus, cytoplasm, and mitochondrion proteins. O-GlcNAcylation is thought to regulate protein function analogously to protein phosphorylation; indeed, crosstalk between both PTMs allows the cell to regulate its functions in response to nutrient levels and stress. Herein, we demonstrate O-GlcNAcylation in T. cruzi epimastigotes by three methods: by using specific antibodies against the modification in lysates and whole parasites, by click chemistry labeling, and by proteomics. In total, 1,271 putative O-GlcNAcylated proteins and six modification sequences were identified by mass spectrometry (data available via ProteomeXchange, ID PXD010285). Most of these proteins have structural and metabolic functions that are essential for parasite survival and evolution. Furthermore, O-GlcNAcylation pattern variations were observed by antibody detection under glucose deprivation and heat stress conditions, supporting their possible role in the adaptive response. Given the numerous biological processes in which O-GlcNAcylated proteins participate, its identification in T. cruzi proteins opens a new research field in the biology of Trypanosomatids, improve our understanding of infection processes and may allow us to identify new therapeutic targets.

Effects on Meccus pallidipennis (Hemiptera: Reduviidae) Eggs Exposed to Entomopathogenic Fungi: Exploring Alternatives to Control Chagas Disease.

Meccus pallidipennis Stål is a vector for Chagas disease. The extensive use of pyrethroid insecticides to control triatomines in Mexico has resulted in the development of resistant populations. As an alternative control approach, the effects on M. pallidipennis eggs of two entomopathogenic fungi, Isaria fumosorosea Wize (Hypocreales: Cordycipitaceae) EH-511/3 and Metarhizium anisopliae (Metschn.) Sorokin (Hypocreales: Clavicipitaceae) EH-473/4, were examined. Egg mortality was estimated 1 mo after egg infection, based on hyphal growth and unsuccessful hatching as proxies for infection and death. Sporulation and conidial production rates were also recorded. Mortality rates caused by I. fumosorosea and Me. anisopliae were 92% ± 3.1 and 88% ± 3.7, respectively. Sporulation rate and conidial production were greater in I. fumosorosea than in Me. anisopliae. Transmission electron microscopy revealed hyphal penetration by both fungal species and damage to embryonic epidermal and cuticular cells. Our results demonstrated that I. fumosorosea and Me. anisopliae are promising candidates for controlling M. pallidipennis eggs and offer alternatives to control the transmission of Chagas disease under natural conditions.

Influence of temperature and humidity on the biology of Triatoma mexicana (Hemiptera: Reduviidae: Triatominae) under laboratory conditions.

Several biological parameters related to the Triatoma mexicana life-cycle were evaluated in this study. Three cohorts were maintained under different combinations of temperature and relative humidity (RH): 25 degrees C/50% RH; 25 degrees C/75% RH; and 30 degrees C/75% RH. Observed hatching rates varied from 49-57.5% whereas the average time of hatching varied from 19.5-22.7 days. In the three cohorts studied, the mean time-lapse between presentation of the blood meal and the beginning of feeding was less than 5 min in all instars; the mean feeding time was longer than 10 min in all the instars; the post-feed defecation delay was over 10 min in all the instars. Less than 50% of nymphs in each cohort completed the cycle and the average time from 1st instar nymph to adult was more than 255 days for the three cohorts. The number of blood meals before molt at each nymphal instar varied from 1-9. Our results appear to indicate a lack of influence of temperature and RH on the biological parameters of T. mexicana that were studied, which could reflect the adaptation capacity of this species. We also conclude that T. mexicana can not be considered an effective transmitter of Trypanosoma cruzi to human populations in areas where this species is currently present.