Effect of Heated Saline Solution on Pain Intensity, Wound Bed Temperature, and Comfort during Chronic Wound Dressing Changes: Crossover Randomized Clinical Trial.

OBJECTIVE: To evaluate the use of heated saline solution during wound cleaning on the intensity of pain related to the procedure, the temperature of the wound bed, and the comfort of patients with chronic wounds. Further, to investigate patient preference in relation to the temperature of the solution used for cleaning. METHODS: Crossover, single-blind, clinical trial with 32 people with chronic wounds. Providers cleaned the wounds with room temperature and heated saline solution. Participants were randomized into group 1 A/B (heated solution first, room temperature second) or group 2 B/A (room temperature solution first, heated solution second), with a 10-minute washout period. Investigators evaluated pain intensity, wound bed temperature, and patient-reported comfort and preference. RESULTS: The heated solution was preferred (P = .04) and more often referred to as comfortable (P = .04) by the participants. There was no difference in pain intensity before and after cleaning with room temperature (2.03; P = .155) and heated saline (2.25; P = .44). The heated solution increased the temperature of the wound bed by 0.5 °C. CONCLUSIONS: Although heating saline solution could be an important comfort measure during dressing changes, quantitatively, the temperature of the solution did not significantly change the temperature of the wound bed nor the intensity of pain patients experienced.

Macroporous latex biomembrane from Hancornia speciosa modulates the inflammatory process and has a debridement effect on wound healing in rats.

PURPOSE: The angiogenic, osteogenic and anti-inflammatory activity of latex of Hancornia speciosa has been evidenced and indicates pharmacological potential with great applicability in the health area, especially in the wound healing process. The present work aimed to compare the effects of the H. speciosa macroporous latex biomembrane with saline on wound healing. METHODS: Forty-three Wistar rats were submitted to excisional wound induction procedure and divided into groups according to treatment: saline (G1), and macroporous biomembrane (G2). The animals were euthanized at three, seven, 14, and 21 days after injury induction (DAI), and three animals were used for the debridement test. Morphometric, macroscopic, and microscopic analyses of general pathological processes were performed. RESULTS: The macroporous biomembrane minimized necrosis and inflammation during the inflammatory and proliferative phases of the healing process, confirmed by the lower intensity of the crust and the debridement effect. In addition, the wounds treated with the macroporous biomembrane presented greater contraction rates in all the experimental periods analyzed. CONCLUSIONS: The macroporous biomembrane presents angiogenic, anti-inflammatory and debridement effects, contributing to the healing process, and can be considered a potentially promising new biomaterial to be used as a dressing.

Albendazole - Ivermectin combination decreases inflammation in experimental neurocysticercosis.

Neurocysticercosis (NCC) is a public health issue in endemic regions and is considered the main preventable cause of neurologic disease. It is caused by the presence of Taenia solium cysticercus in the central nervous system. The current treatment is performed with anthelminthic drugs - albendazole (ABZ) or praziquantel - associated with anti-inflammatory and corticosteroids in order to prevent the negative effects of the inflammatory reaction to the parasite's death. Ivermectin (IVM) is an anthelminthic drug that has been shown to present an anti-inflammatory effect. The aim of this study was to was to evaluate the histopathologic aspects of experimental NCC after in vivo treatment with a combination of ABZ-IVM. Balb/c mice were intracranially inoculated with T. crassiceps cysticerci and after 30 days of infection were treated with a single dose of NaCl 0.9% (control group), ABZ monotherapy (40 mg/kg), IVM monotherapy (0.2 mg/kg) or a combination of ABZ-IVM. 24h after the treatment the animals were euthanized and the brain was removed for histopathologic analysis. The IVM monotherapy and ABZ-IVM combination showed more degenerated cysticerci, less inflammatory infiltration, meningitis and hyperemia than the other groups. Therefore, it is possible to recommend the combination of albendazole and ivermectin as alternative chemotherapy for NCC due to its antiparasitic and anti-inflammatory effects, with potential to decrease the negative effects of the inflammatory burst when the parasite is killed within the CNS.

Polymethylmethacrylate Microspheres are Immunologically Inert in Mouse Tissues.

Nowadays, aesthetic concerns have gained attention, especially by patients looking for a less invasive alternative to minor facial corrections. Polymethylmethacrylate (PMMA) is widely used as a soft tissue filler; the demand for this polymer has increased, and along with it, there are some reports of adverse reactions. Such adverse reactions stem from consequences of immune and inflammatory reactions to PMMA. Some animal models have been used to unravel the causes of these reactions, among other factors involving the management of PMMA. The aim of this study was to determine the immunogenic profile of PMMA implantation in different anatomical planes of mice, over up to 360 experimental days. In this study, BALB/c mice were divided into 30 groups for immune evaluation of the interaction between the organism and the polymer; 2% PMMA was implanted subcutaneously, 10% intramuscularly and 30% in periosteal juxtaposition and followed during five experimental days (7, 30, 90, 180 and 360 days after implantation-DAI). Pro- and anti-inflammatory cytokines (IL-2, IL-4, IL-6, IFN-gamma, TNF, IL-17A, IL-10 and TGF-beta) were quantified in all experimental days. There was no statistical difference between the groups analyzed considering the evaluated parameters. Therefore, at all implanted depths, PMMA behaved inertly in a murine model.No Level Assigned This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Tissue Remodeling After Implantation with Polymethylmethacrylate: An Experimental Study in Mice.

Polymethylmethacrylate (PMMA) is a filler used for aesthetic and/or repair purposes. The response to the implantation of biomaterials varies according to factors related to the patient, the professional responsible for the application and the material used. In vitro and in vivo experimental models have been used to study aspects such as the organism/biomaterial interface and the role of macrophages, dendritic cells and neutrophils. This study aimed to characterize the inflammatory reactions related to polymer concentration, implantation depth and exposure time. Different concentrations of PMMA were implanted in different anatomical planes in mice. The consequences of contact with PMMA, from structural changes to the inflammatory characteristic of tissue damage, were histologically evaluated. The implantation interfered in the morphological structure of the region where it was implanted, expanding it and due to the inflammatory reaction generated, by the presence of the vehicle in the initial phase and by the collagen produced in the chronic phase. The 30% concentration of PMMA induced a greater presence of foreign body giant cells both subcutaneously, at 7, 30 and 90 days after implantation (DAI), and intramuscular at 30DAI. Tissue remodeling was more expressive in the subcutaneous region with significant density of the extracellular matrix at 90DAI. In conclusion, the foreign body reaction resulting from the implantation process acquires different characteristics depending on the anatomical plane and the concentration of implanted product, where the more superficial the implantation plane, the greater the inflammatory reaction. Moreover, PMMA concentration and the depth of implantation did not influence the collagen production.No Level Assigned This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors https://www.springer.com/00266.

Macroporous latex biomembrane from Hancornia speciosa modulates the inflammatory process and has a debridement effect on wound healing in rats

Purpose: The angiogenic, osteogenic and anti-inflammatory activity of latex of Hancornia speciosa has been evidenced and indicates pharmacological potential with great applicability in the health area, especially in the wound healing process. The present work aimed to compare the effects of the H. speciosa macroporous latex biomembrane with saline on wound healing. Methods: Forty-three Wistar rats were submitted to excisional wound induction procedure and divided into groups according to treatment: saline (G1), and macroporous biomembrane (G2). The animals were euthanized at three, seven, 14, and 21 days after injury induction (DAI), and three animals were used for the debridement test. Morphometric, macroscopic, and microscopic analyses of general pathological processes were performed. Results: The macroporous biomembrane minimized necrosis and inflammation during the inflammatory and proliferative phases of the healing process, confirmed by the lower intensity of the crust and the debridement effect. In addition, the wounds treated with the macroporous biomembrane presented greater contraction rates in all the experimental periods analyzed. Conclusions: The macroporous biomembrane presents angiogenic, anti-inflammatory and debridement effects, contributing to the healing process, and can be considered a potentially promising new biomaterial to be used as a dressing.

Treatment of experimentally induced partial-thickness burns in rats with different silver-impregnated dressings.

PURPOSE: To evaluate the morphometric, macroscopic and microscopic aspects of experimentally induced partial-thickness burns in rats treated with different silver-based dressings. METHODS: Wistar rats were used, divided into six treatments: saline (NaCl 0.9%); silver sulfadiazine 1%; Silvercel; Mepilex Ag; Aquacel Ag and Acticoat. The animals were monitored daily and euthanized at 7, 14 and 30 days after injury induction (DAI). RESULTS: At 7 DAI, necrosis/crust was greater in control, silver sulfadiazine and Mepilex Ag treatments, granulation tissue was induced by Aquacel Ag, polymorphonuclear infiltrate (PMN) infiltration was intensified by Mepilex Ag; mononuclear infiltrate (MN) infiltration and angiogenesis were increased by Silvercel. At 14 DAI, hemorrhage was decreased by Silvercel and Mepilex Ag, PMN infiltration increased by Acticoat. At 30 DAI, angiogenesis was greater in the Acticoat treatment and fibroblasts were increased by Acticoat and Mepilex Ag. Collagen was induced at 14 DAI by silver sulfadiazine and Aquacel Ag and, at 30 DAI, by silver sulfadiazine and Silvercel treatments. CONCLUSIONS: Silvercel and Acticoat presented better results than the other products. However, all the dressings were better than the control at some point during the process, and may contribute to the healing of partial thickness burns. Silvercel and Aquacel Ag treatments induced better cosmetic outcomes regarding wound closure and scarring.

Hydrogel-based dressings in the treatment of partial thickness experimentally induced burn wounds in rats.

PURPOSE: To compare four commercially available hydrogel formulations in the healing of partial thickness burns experimentally induced in rats. METHODS: Wistar rats were used, and after the burn wound induction they were divided into the following treatment groups: G1) NaCl 0.9%; G2) 1% silver sulfadiazine; G3) Debrigel™; G4) Safgel™; G5) Dersani™; G6) Solosite™. The animals were followed during seven, 14 and 30 days after the injury induction. Morphometric, macroscopic and microscopic evaluations were performed. RESULTS: The treatment with Dersani™ induced better results during the inflammatory and proliferative phases of the healing process (p<0.05). The animals treated with Safgel™ presented better scaring in the remodeling phase (p<0.05), and the treatment with Dersani™ and Solosite™ induced greater wound closure (p<0.05). CONCLUSIONS: The hydrogel-based dressings presented beneficial outcomes in the healing of burn wounds experimentally induced in rats due to their ability in maintain the humidity of the wound, in removing the exudate, in promoting cell migration and collagen production during the different phases of the healing process.

In vitro metabolic stress induced by nitazoxanide and flubendazole combination in Taenia crassiceps cysticerci.

Taenia crassiceps is often used as experimental model for T. solium cysticercosis studies. Currently cysticercosis antiparasitic treatment is based on albendazole and praziquantel which may present side effects and parasitic resistance. The search for other antiparasitic drugs is necessary. Nitazoxanide (NTZ) and flubendazole (FLB) are broad spectrum antiparasitic drugs that present anti-cysticercosis effect. Metabolic analyses help to determine the impact of these drugs on parasites. The aim of this study was to determine the impact on the production and excretion of organic metabolites in T. crassiceps cysticerci after in vitro exposure to NTZ and FLB, isolated or in combination. T. crassiceps cysticerci were culture in RPMI medium and exposed to 10 µg/mL of NTZ, 10 µg/mL of FLB or 10 µg/mL of NTZ +10 µg/mL of FLB. 24 h after exposure, the parasites were chromatographic analyzed to determine the impact of these drugs on glycolysis, homolactic fermentation, tricarboxylic acid cycle, fatty acids oxidation and proteins catabolism. It was possible to determine that the drugs combination induced greater metabolic impact on cysticerci in comparison to the isolated drugs exposure. The drugs combination induced gluconeogenesis, metabolic acidosis, increase in tricarboxylic acid cycle and in proteins catabolism. While the NTZ isolated exposure induced metabolic acidosis and protein catabolism and the FLB isolate exposure induced gluconeogenesis and protein catabolism. These results show that the combination of drugs with different modes of action increase the antiparasitic effect and may be indicated as alternative cysticercosis treatments.

Intraperitoneal and intracranial experimental cysticercosis present different metabolic preferences after treatment with isolated or combined albendazole and nitazoxanide.

Cysticercosis is a zoonotic public health issue especially severe when the parasite is in the central nervous system although it may be found all over the human organism. Taenia crassiceps cysticerci inoculated in mice is the experimental model used to study cysticercosis. The most used cysticercosis treatment is with albendazole (ABZ). Nitazoxanide (NTZ) has been experimentally tested against this parasite. Metabolic analysis has been used to determine drugs impact on the parasite. The aim of this study was to determine the in vivo metabolic impact of the ABZ-NTZ combination in T. crassiceps cysticerci inoculated in mice peritoneal and intracranial cavities. Mice were experimentally inoculated with T. crassiceps cysticerci in the intraperitoneal cavity or in the intracranial one. Thirty days after the infection they were treated with NaCl 0.9% (control group), 50 mg/kg of ABZ, 50 mg/kg of NTZ or 50 mg/kg of NTZ and ABZ (ABZ/NTZ combination). 24 h after treatment the animals were euthanized and the cysticerci analyzed through high performance chromatography and spectrophotometry in order to detect the glycolytic, mitochondrial and protein catabolism pathways. The intracranial parasites used more intensely the homolactic fermentation while the intraperitoneal ones presented a greater use of the mitochondrial pathways and protein catabolism. Regarding the glycolytic pathways, it was possible to observe a significant impact induced by the drugs used, both isolated or in combination. It was possible to detect an increase in the fumarate reductase pathway after the drugs exposure and no impact in the protein's catabolism. Therefore, the cysticerci showed different uses of metabolic pathways regarding the site of inoculation due to the availability of nutrients inherent of each environment. This study showed the parasite metabolic resilience and capability of use of different biochemical pathways in order to ensure survival in spite of a hostile environment.

Treatment of experimentally induced partial-thickness burns in rats with different silver-impregnated dressings

Purpose: To evaluate the morphometric, macroscopic and microscopic aspects of experimentally induced partial-thickness burns in rats treated with different silver-based dressings. Methods: Wistar rats were used, divided into six treatments: saline (NaCl 0.9%); silver sulfadiazine 1%; Silvercel; Mepilex Ag; Aquacel Ag and Acticoat. The animals were monitored daily and euthanized at 7, 14 and 30 days after injury induction (DAI). Results: At 7 DAI, necrosis/crust was greater in control, silver sulfadiazine and Mepilex Ag treatments, granulation tissue was induced by Aquacel Ag, polymorphonuclear infiltrate (PMN) infiltration was intensified by Mepilex Ag; mononuclear infiltrate (MN) infiltration and angiogenesis were increased by Silvercel. At 14 DAI, hemorrhage was decreased by Silvercel and Mepilex Ag, PMN infiltration increased by Acticoat. At 30 DAI, angiogenesis was greater in the Acticoat treatment and fibroblasts were increased by Acticoat and Mepilex Ag. Collagen was induced at 14 DAI by silver sulfadiazine and Aquacel Ag and, at 30 DAI, by silver sulfadiazine and Silvercel treatments. Conclusions: Silvercel and Acticoat presented better results than the other products. However, all the dressings were better than the control at some point during the process, and may contribute to the healing of partial thickness burns. Silvercel and Aquacel Ag treatments induced better cosmetic outcomes regarding wound closure and scarring.

Alterations in Taenia crassiceps cysticerci cytoskeleton induced by nitazoxanide and flubendazole.

Cysticercosis is the presence of Taenia solium larval stage in tissues such as central nervous system, skin, muscles and eye globe. The current treatment is based on albendazole and praziquantel which already present resistance reports. Therefore, the search for alternative treatments is paramount. The aim of this study was to determine the effect of flubendazole and nitazoxanide on cytoskeleton proteins from Taenia crassiceps cysticerci, an experimental model for cysticercosis. Cysticerci were cultured in RPMI supplemented medium containing nitazoxanide and/or flubendazole. 24 h after the exposure the cysticerci were processed for scanning and transmission electron microscopy and for protein analysis of the cytoskeleton. The proteins were detected through 1D electrophoresis and identified through Western Blot. Nitazoxanide exposure increased tubulin and actin quantifications in T. crassiceps cysticerci. While flubendazole alone and the drugs combinations induced an increase in α-tubulin and actin and decreased ß-tubulin quantifications in the parasite. Morphological changes such as swelling and rupture of vesicle, stiff membrane, decrease in movements were observed when the cysticerci were incubated with the different compounds. In conclusion the drugs induced significative impact in the parasite`s cytoskeleton and may be considered as alternative treatments for cysticercosis.

Molecular characterization of Toxoplasma gondii isolates from free-range chickens reveals new genotypes in Goiânia, Goiás, Brazil.

The aim of this study was to evaluate the genotypic characteristics of Toxoplasma gondii isolated from free-range chickens in the metropolitan area of Goiânia, Goiás, in Brazil's central-west region. The seroprevalence rate was found to be 96%, according to an indirect hemagglutination assay. Brain and heart samples were processed by peptic digestion for a mice bioassay. The tissues were homogenized and the resulting samples were subjected to polymerase chain reaction (PCR), which revealed that 64% of them contained the parasite's DNA. The mice bioassay revealed 15 isolates, 8 of them tachyzoites isolates from the peritoneal lavage and 7 from brain cysts. T. gondii genotypes were determined through PCR-RFLP, using the following markers: SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, alt. SAG2, Apico and CS3. Three genotypes were identified, inclued ToxoDB #65, and the other two are not yet described in the literature. Hence, we conclude that the isolates obtained from the metropolitan area of Goiânia showed relatively low genetic diversity.

Carboxymethyl chitosan hydrogel formulations enhance the healing process in experimental partial-thickness (second-degree) burn wound healing.

PURPOSE: This study aimed to elaborate a hydrogel constituted by carboxymethyl chitosan (CMC), hyaluronic acid (HA) and silver (Ag) and to evaluate its healing effect on partial-thickness burn wounds experimentally induced in rats. METHODS: CMC was obtained by chitosan reacting with monochloroacetic acid. The carboxymethylation was confirmed by Fourier-transform infrared spectroscopy and hydrogen nuclear magnetic resonance (NMR). Scanning electron microscopy was used to determine the morphologicalcharacteristics of chitosan and CMC. After the experimental burn wound induction, the animals (n = 126) were treated with different CMC formulations, had their occlusive dressings changed daily and were followed through 7, 14 and 30 days. Morphometric, macroscopic and microscopic aspects and collagen quantification were evaluated. RESULTS: Significative wound contraction, granulation tissue formation, inflammatory infiltration and collagen fibers deposit throughout different phases of the healing process were observed in the CMC hydrogels treated groups. CONCLUSIONS: The results showed that, in the initial phase of the healing process, the most adequate product was the CMC/HA/Ag association, while in the other phases the CMC/HA association was the best one to promote the healing of burn wounds.

Polymethyl Methacrylate (PMMA) in the Treatment of a Case of Hemifacial Microsomia.

Hemifacial microsomia (HMF) is a morphological alteration characterized by facial deformities. These alterations are a consequence of a congenital anomalous development of the first and second branchial arches. It may present a genetic or environmental origin or a mixture of both and is considered mostly multifactorial. The clinical presentation varies; however, some characteristics are predominant such as unilateral mandible hypoplasia, agenesis or malformation of the auricular pavilion, and agenesis or malformation of the eye globe. The aim of this paper was to report a clinical case of a patient with late diagnosis of HMF which presented multiple deformities and received treatment with polymethyl methacrylate (PMMA) implant in different concentrations. Aiming for facial harmony, the PMMA implant occurred in the following regions: canine fossa, nasolabial sulcus, mandible, Bichat Ball, chin outline, lip contour, nose base, columella, nose tip, and dorsum. The treatment resulted in significant improvement in the facial symmetry.

Partial inhibition of the main energetic pathways and its metabolic consequences after in vivo treatment with benzimidazole derivatives in experimental neurocysticercosis.

Benzimidazole derivatives such as albendazole (ABZ) and mebendazole are important molecules used in helminthic treatment. Neurocysticercosis is the main cause of acquired epilepsy throughout the world and is currently treated with ABZ. New molecules have been studied in order to aid in the treatment of this neglected tropical disease, among them RCB15 and RCB20. The aim of this study was to evaluate the metabolic impact of RCB15 and RCB20 on Taenia crassiceps cysticerci intracranially inoculated in Balb/c mice. Thirty days after the inoculation the mice were treated with 50 mg kg-1 of RCB15, RCB20, ABZ or NaCl 0.9%. The euthanasia and cysticerci removal were performed 24 h after the treatment. The cysticerci were analysed through high performance liquid chromatography. After the treatments, there was an impairment in the main energetic pathways such as glycolytic pathway, homolactic fermentation or in mitochondrion energy production detected through the decrease in pyruvate, lactate, oxaloacetate, malate and fumarate concentrations. This induced the parasite to resort to alternative energetic pathways such as proteins catabolism, propionate fermentation and fatty acids oxidation. Therefore, benzimidazole derivatives are a promising alternative to ABZ use as they also reach the brain tissue and induce a metabolic stress in the cysticerci.

The first Acanthamoeba keratitis case in the Midwest region of Brazil: diagnosis, genotyping of the parasite and disease outcome.

We report an Acanthamoeba keratitis case associated with the use of contact lens in a 28-year-old female from Brasília, Brazil. Samples from corneal scraping and contact lens case were used for culture establishment, PCR amplification, and partial sequencing (fragments of ~400kb) of small subunit rDNA; both culture and PCR were positive. The sequence analyses of the cornea and of isolates from the contact lens case showed similarity with the T4 genotype. To the best of our knowledge, this is the first report of T4 Acanthamoeba keratitis case from the Midwest region of Brazil.

The first Acanthamoeba keratitis case in the Midwest region of Brazil: diagnosis, genotyping of the parasite and disease outcome

Abstract We report an Acanthamoeba keratitis case associated with the use of contact lens in a 28-year-old female from Brasília, Brazil. Samples from corneal scraping and contact lens case were used for culture establishment, PCR amplification, and partial sequencing (fragments of ~400kb) of small subunit rDNA; both culture and PCR were positive. The sequence analyses of the cornea and of isolates from the contact lens case showed similarity with the T4 genotype. To the best of our knowledge, this is the first report of T4 Acanthamoeba keratitis case from the Midwest region of Brazil.

Facial trauma reconstruction with polymethyl methacrylate-A case report.

BACKGROUND: Polymethyl methacrylate (PMMA) is a dermatologic filler commonly used in esthetic procedures. However, it can also be used in more severe cases such as reconstruction of facial traumas. AIMS: The aim of this report was to describe the use of PMMA in a nasal reconstruction of a patient victim of an automobilist accident which presented previous rejection of autograft. PATIENT: A young female patient, 26 y, victim of automobilist accident, with no bone support due to trauma both in the mandibulum and maxilla, presented fracture of several facial bones. Presented rejection of a bone autograft from the skullcap implanted on the nose. In order to harmonize the facial aspect, a nasal fill with PMMA for the anatomic reconstruction of the nose was performed. RESULTS AND CONCLUSIONS: PMMA promoted the recovery of facial and nasal esthetic characteristics of the patient ensuring a satisfactory result.

Promastigote parasites cultured from the lesions of patients with mucosal leishmaniasis are more resistant to oxidative stress than promastigotes from a cutaneous lesion.

Human leishmaniasis caused by Leishmania (Viannia) braziliensis can be presented as localized cutaneous leishmaniasis (LCL) or mucosal leishmaniasis (ML). Macrophages kill parasites using nitric oxide (NO) and reactive oxygen species (ROS). The aim of this study was to evaluate the ability of parasites obtained from patients with LCL or ML to produce and resist NO or ROS. Promastigotes and amastigotes from LCL or ML isolates produced similar amounts of NO in culture. Promastigotes from ML isolates were more resistant to NO and H2O2 than LCL parasites in a stationary phase, whereas amastigotes from LCL isolates were more resistant to NO. In addition, in the stationary phase, promastigote isolates from patients with ML expressed more thiol-specific antioxidant protein (TSA) than LCL isolates. Therefore it is suggested that infective promastigotes from ML isolates are more resistant to microbicidal mechanisms in the initial phase of infection. Subsequently, amastigotes lose this resistance. This behavior of ML parasites can decrease the number of parasites capable of stimulating the host immune response shortly after the infection establishment.