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Chagas disease, caused by the protozoan Trypanosoma cruzi, represents an important and worldwide public health issue, particularly in Latin America. Limitations of conventional treatment with benznidazole and nifurtimox underscore the urgent need for new therapeutic strategies for this disease. Schinus molle, a tree used in traditional medicine for various ailments, has demonstrated promising antiparasitic activity. The in vitro anti-T. cruzi activity of Schinus molle crude methanol extract, partitions, and fractions, as well as their cytotoxicity in Vero cells and Artemia salina, and hemolytic activity in human erythrocytes were assessed. Most of the extracts possessed anti-T. cruzi effects, with Sm-CF3 being the fraction with the highest activity (IC50 = 19 µg/mL; SI = 6.8). Gas chromatography–mass spectrometry analysis identified 20 compounds, with fatty acyls comprising the predominant chemical class (55%). We also identified the antiparasitic compounds cis-5,8,11,14,17-eicosapentaenoic acid and trans-Z-α-bisabolene epoxide, suggesting their potential contribution to the observed anti-T. cruzi activity. In conclusion, our findings support the therapeutic potential of S. molle as a source of novel antiparasitic agents against T. cruzi. [ABSTRACT FROM AUTHOR]

Beauveria bassiana has potential for Aedes aegypti biological control. However, its efficacy depends on the strain's geographic location, host susceptibility, and virulence. The present study aimed to evaluate the effectiveness of B. bassiana strain BBPTG4 conidia in controlling Ae. aegypti adults and its detection via introns profile on exposed mosquito corpses. Morphologic characteristics among strains were highly similar. Comprehensive testing of these strains demonstrated that BBPT4 exhibited the ideal biological activity for Ae. aegypti control, with a median lethal time (TL50) of 7.5 d compared to ~3 d and ~10 d for BB01 and BB37 strains, respectively. Infected mosquitoes died after GHA and BBPTG4 exposure, and corpses were analyzed for infecting strains detection. Differences among the seven evaluated strains were determined, assessing five different insertion group I intron profiles in BBTG4, BB01, GHA, BB37, and BB02 strains. Mosquitoes infected by BBPTG4 and non-exposed (negative control) intron profiles were obtained. We detected the presence of introns in the BBPTG4 strain, which were not present in non-exposed mosquitoes. In conclusion, B. bassiana strains showed similarities in terms of their cultural and microscopic morphological characteristics and biologicals virulence level, but different intron profiles. BBPTG4 strain-infected Ae. aegypti adult corpses, showing specific amplicons, enabled us to identify B. bassiana at the strain level among infected mosquitoes. However, monitoring and detection of field-infected insects is essential for further verification. [ABSTRACT FROM AUTHOR]

Rotavirus is the main cause of acute diarrhea in children up to five years of age. In this regard, probiotics are commonly used to treat or prevent gastroenteritis including viral infections. The anti-rotavirus effect of Bifidobacterium longum and Chlorella sorokiniana, by reducing viral infectivity and improving IFN-type I response, has been previously reported. The present study aimed to study the effect of B. longum and/or C. sorokiniana on modulating the antiviral cellular immune response mediated by IFN-γ, IL-10, SOCS3, STAT1, and STAT2 genes in rotavirus-infected cells. To determine the mRNA relative expression of these genes, HT-29 cells were treated with B. longum and C. sorokiniana alone or in combination, followed by rotavirus infection. In addition, infected cells were treated with B. longum and/or C. sorokiniana. Cellular RNA was purified, used for cDNA synthesis, and amplified by qPCR. Our results demonstrated that the combination of B. longum and C. sorokiniana stimulates the antiviral cellular immune response by upregulating IFN-γ and may block pro-inflammatory cytokines by upregulating IL-10 and SOCS3. The results of our study indicated that B. longum, C. sorokiniana, or their combination improve antiviral cellular immune response and might modulate pro-inflammatory responses. [ABSTRACT FROM AUTHOR]

Beauveria bassiana (B. bassiana) is a significant entomopathogenic fungus (EPF) in agriculture as a sprayable biocontrol agent. It has the potential to be established as an endophyte (ENP) in various crops, resulting in beneficial effects for the host plants, including resistance to pest insects and increased growth and yield. However, it is not known whether a B. bassiana strain has such a favorable impact on the plant, since it is a common soil microorganism. Therefore, techniques that allow strain monitoring will be advantageous. To date, methods for detecting or monitoring a specific EPF strain after external application are scarce. In the present study, an in planta nested PCR technique was standardized to differentiate between three B. bassiana strains (GHA, PTG4, and BB37) established as endophytes in bean plants under laboratory conditions by detecting the insertion profile of four group I introns located in the 28S gene of B. bassiana ribosomal DNA. This technique recognized a distinct pattern of bands of different sizes for each strain, with a sensitivity of 1 pg per 10 ng of plant DNA. This molecular approach may be more effective monitoring B. bassiana strains after application to evaluate their significance on crops. [ABSTRACT FROM AUTHOR]

Hirsutella citriformis Speare is the only entomopathogenic fungus that has been applied to control the hemipteran Diaphorina citri Kuwayama. However, the use of available commercial products under field conditions is limited due to conidia's shelf life and short environmental persistence. We have previously reported the citrus psyllid D. citri adults' biocontrol potential using H. citriformis strains. The aim of the present study was to evaluate different formulations based on H. citriformis (OP-Hir-3, OP-Hir-10, and OP-Hir-12 strains) conidia and gums as additives to improve D. citri adults' biocontrol, under laboratory, greenhouse, and field conditions, using Hirsutella gums as conidia stabilizers to improve their viability under environmental drought conditions and as insecticide. Laboratory bioassay results showed that the highest (p < 0.05) D. citri mortality was achieved using FOP-Hir-10GH (63.5%), followed by the Hirsutella gum control (42.2%). Under greenhouse conditions, adults' mortality reached up to 84.6% with FOP-Hir-12 and 49.0% with Hirsutella gum. In addition, we applied H. citriformis formulations under field conditions in a commercial citrus grove located in Tecomán, Colima, México, at 21.5 °C and 73.3% relative humidity (RH) in March and 25.7 °C and 72.5% RH in October 2022 and observed 67.3% and 94.0% mortality of D. citri adults, respectively. Hirsutella gum alone showed significant insecticidal activity against D. citri adults. In conclusion, this study demonstrated that Hirsutella gum functioned as additive to H. citriformis conidia formulations, improving D. citri adults' mortality and showing potential for this pest biocontrol in citrus orchards. [ABSTRACT FROM AUTHOR]

Biological control by beneficial microorganisms is known to significantly reduce the effect of pests on crops yield. Among the biocontrol strategies is the use of entomopathogenic fungi such as Hirsutella citriformis, which has been applied to infect and kill hemipteran insect pests, including Diaphorina citri Kuwayama (Hemiptera: Liviidae) and Bactericera cockerelli Sulc. (Hemiptera: Triozidae). These biological agents are applied in the form of conidia that are often combined with other inert materials to facilitate application, protect conidia, and improve their shelf life. The aim of this study was to implement strategies for developing formulations to increase conidia shelf life. We evaluated gum produced from one strain and conidia from two different H. citriformis strains. Conidia were formulated by evaluating different concentrations of Acacia and Hirsutella gums to enhance conidia viability during storage at 4 °C or 25 °C. Results indicated that formulations maintained conidia viability for at least 90 d after storage at 25 °C (≥70% viability) and at least 120 d after storage at 4 °C, which was significantly (p ≤ 0.05) higher than that of the control, without observing changes in pH values. We also demonstrated 100% formulation purity from days 0 to 120, among all treatments. In conclusion, evaluated formulations maintained H. citriformis conidia viability for at least three months, when stored at 4 °C. [ABSTRACT FROM AUTHOR]

Ruta chalepensis is an herb used to treat various ailments, and its potential cytotoxic effects on different tumor cell lines have been extensively studied. The present study aimed to evaluate the cytotoxic activity of R. chalepensis methanol extract (RCME), sub-partitions obtained from solvents of increasing polarity, and major compounds, as well as their hemolytic, anti-hemolytic, and antioxidant potential. The in vitro cytotoxic activity against the human hepatocarcinoma (HEP-G2) and the murine lymphoma cell line (L5178Y-R) was evaluated using the colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay, whereas selectivity indices (SIs) were determined by comparing cytotoxicity against normal African green monkey kidney cells (VERO) and human peripheral blood mononuclear cells (PBMC). Hemolytic and anti-hemolytic activities were evaluated on human erythrocytes. The most effective cytotoxic treatment was evaluated for nitric oxide release by J774A.1 macrophages. Antioxidant activity of R. chalepensis material was also determined. Results showed that RCME produced significant (p < 0.05) cytotoxicity in HEP-G2 (IC50 = 1.79 µg/mL) and L5178Y-R (IC50 = 1.60 µg/mL) cells and exhibited high SIs (291.50 and 114.80, respectively). In addition, the n-hexane fraction (RCHF) showed an IC50 of 18.31 µg/mL in HEP-G2 cells and an SI of 9.48 in VERO cells, whereas the chloroform fraction (RCCF) evidenced an IC50 of 1.60 µg/mL in L5178Y-R cells and an SI of 34.27 in PBMC cells. Chalepensin (CHL), rutamarin (RTM), and graveolin (GRV), which are major components of R. chalepensis, showed high activity against L5178Y-R cells, with IC50 of 9.15, 15.13 and SI of 45.08 µg/mL, respectively. In addition, CHL, RTM, and GRV showed SIs of 24.76, 9.98, and 3.52, respectively, when compared with PBMC cells. RCME at concentrations of 125 µg/mL and 250 µg/mL, significantly (p < 0.05) decreased nitrite production in J774A.1 cells, when exposed to lipopolysaccharide. This study demonstrated that RCME showed significant cytotoxic activity against HEP-G2 and L5178Y-R cells, without affecting normal VERO, PBMC, and J774A.1 cells. [ABSTRACT FROM AUTHOR]

Probiotics are effective to treat or prevent gastrointestinal infections, and microalgae have demonstrated important health-promoting effects and in some cases function as prebiotics. In this regard, the anti-rotavirus effect of Bifidobacterium longum and Chlorella sorokiniana by reducing viral infectivity is well known. However, their effect on immune response against rotavirus has not yet been investigated. Therefore, the aim of this study was to determine the role of Bifidobacterium longum and/or Chlorella sorokiniana in influencing an IFN type I-mediated antiviral response in rotavirus-infected cells. In pre-infection experiments, HT-29 cells were treated with B. longum and C. sorokiniana alone or in combination, followed by rotavirus infection, whereas in post-infection assays, HT-29 cells were treated after infection. The cells' mRNA was then purified to determine the relative expression level of IFN-α, IFN-β, and precursors of interferons such as RIG-I, IRF-3, and IRF-5 by qPCR. We showed that combination of B. longum and C. sorokiniana significantly increased IFN-α levels in pre-infection and IFN-β in post-infection assays, as compared with individual effects. Results indicate that B. longum, C. sorokiniana, or their combination improve cellular antiviral immune response. [ABSTRACT FROM AUTHOR]

Cancer is a major health problem with significant morbidity and mortality. In addition, plants are a source of metabolites with diverse biological properties, including antitumor potential. In this study, we investigated the in vitro murine lymphoma L5178Y-R cell growth inhibition, human peripheral blood mononuclear cells (PBMC) toxicity and proliferation, and antioxidant, hemolytic, and anti-hemolytic activities of methanol extracts from 15 plants of traditional use in Mexico. Justicia spicigera caused the highest tumor cell growth inhibition with a half maximal inhibitory concentration (IC50) of 29.10 µg/mL and a selectivity index >34.36 compared with those of PBMC, whereas Mimosa tenuiflora showed the highest lymphoproliferative activity from 200 µg/mL compared with that induced by concanavalin A. In addition, M. tenuiflora showed an antioxidant effect (IC50 = 2.86 µg/mL) higher than that of ascorbic acid. Regarding the hemolytic and anti-hemolytic activity, all extracts presented significant anti-hemolytic activity. The extract of J. spicigera is emerging as a possible source of effective antineoplastic compounds. [ABSTRACT FROM AUTHOR]

Hirsutella citriformis Speare is the only entomopathogenic fungus involved in Diaphorina citri Kuwayama natural epizootics. The aim of the present study was to evaluate different protein sources as supplements to stimulate Hirsutella citriformis growth, improve conidiation on solid culture, and evaluate its produced gum for conidia formulation against D. citri adults. Hirsutella citriformis INIFAP-Hir-2 strain was grown on agar media enriched with wheat bran, wheat germ, soy, amaranth, quinoa, and pumpkin seed, in addition to oat with wheat bran and/or amaranth. The results demonstrated that 2% wheat bran significantly (p < 0.05) promoted mycelium growth. However, 4% and 5% wheat bran achieved the highest conidiation (3.65 × 107 conidia/mL and 3.68 × 107 conidia/mL, respectively). Higher conidiation (p < 0.05) was observed on oat grains supplemented with wheat bran, as compared with culturing on oat grains without supplements (7.25 × 107 versus 5.22 × 107 conidia/g), after a 14 d instead of 21 d incubation period. After supplementing synthetic medium or oat grains with wheat bran and/or amaranth, INIFAP-Hir-2 conidiation increased, whereas production time was reduced. After using Acacia and Hirsutella gums to formulate conidia produced on wheat bran and amaranth at 4%, field trial results showed that the highest (p < 0.05) D. citri mortality was achieved by Hirsutella gum-formulated conidia (80.0%), followed by the Hirsutella gum control (57.8%). Furthermore, Acacia gum-formulated conidia caused 37.8%, whereas Acacia gum and negative controls induced 9% mortality. In conclusion, Hirsutella citriformis gum used to formulate its conidia improved biological control against D. citri adults. [ABSTRACT FROM AUTHOR]

Medicinal plants are traditionally used in Mexico to treat diseases such as cancer. The present study aimed to evaluate the cytotoxic, antioxidant, and anti-hemolytic activity of 15 plants of ethnopharmacological use in Mexico. For this, plant methanol extracts were prepared by the Soxhlet method, after which their cytotoxic activity was evaluated against human hepatocellular carcinoma (HEP-G2) and monkey kidney epithelial (Vero) cells by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction colorimetric assay. The selectivity index (SI) of each extract was then determined by the IC50 ratio of normal to tumor cells. We showed that Ruta chalepensis extract possessed an IC50 of 1.79 µg/mL and 522.08 µg/mL against HEP-G2 and Vero cells, respectively, resulting in an SI of 291.50. Furthermore, antioxidant activity was evaluated by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging technique, where the best antioxidant potential was shown by the Heterotheca inuloides extract (IC50 = 19.24 µg/mL). Furthermore, the hemolytic potential was determined against human erythrocytes, which showed that the extracts with the highest anti-hemolytic activity were Smilax aspera (IC50 = 4.41 µg/mL) and Amphipterygium adstringens (IC50 = 5.35 µg/mL). In conclusion, we observed that R. chalepensis methanol extract possesses cytotoxic activity against HEP-G2 cells, without affecting non-tumorigenic Vero cells. Our results indicated the antitumor potential of medicinal plants used in Mexico. [ABSTRACT FROM AUTHOR]

Aedes aegypti (Linn.) incidence has increased in recent years, causing human viral diseases such as dengue, which are often fatal. Beauveria bassiana (Bals., Vuillemin) efficacy for Ae. aegypti biological control has been evidenced but it relies on host susceptibility and strain virulence. We hypothesized that B. bassiana conidia microgranular formulations (MGF) with the additives acetone, lactic acid, and sugar increase Ae. aegypti adult exposure, thus improving their biocontrol effectiveness. Beauveria bassiana strain four (BBPTG4) conidia stability was assessed after 0 d, 5 d, and 30 d storage at 25 °C ± 2 °C with additives or in MGF after 91 d of storage at 25 °C ± 2 °C or 4 °C ± 1 °C, whereas mortality was evaluated after adult exposure to MGF + conidia, using home-made traps. Additives did not show toxicity to conidia. In addition, we observed that sugar in MGF increased Ae. aegypti adults' attraction and their viability resulted in a 3-fold reduction after 5 d and 1- to 4-fold decrease after 30 d of storage, and formulations were less attractive (p < 0.05). Conidia stability was higher on MGF regardless of the storage temperature, losing up to 2.5-fold viability after 91 d. In conclusion, BBPTG4 infected and killed Ae. aegypti, whereas MGF attracting adults resulted in 42.2% mortality, increasing fungus auto dissemination potential among infected surviving adults. It is necessary to further evaluate MGF against Ae. aegypti in the field. [ABSTRACT FROM AUTHOR]

Studies on Hirsutella citriformis Speare are scarce. Among these, some reports have focused on phenotypic identification, based on its morphological structure and morphometric characteristics. This fungus is known to control economically important citrus crop pests. In recent years, H. citriformis has received increased attention as a control agent for the Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Liviidae), which causes the Huanglongbing (HLB) disease. Unfortunately, formal H. citriformis strains characterization is marginal, which mainly involves the role of biologically active exudates (metabolites) produced during their growth. Information regarding their mode of action and biocontrol potential is limited. However, epizootics reports of this fungus, under suitable environmental conditions for its development (25 °C to 28 °C and ~80% relative humidity), have demonstrated its parasitization efficacy. Therefore, it becomes challenging to determine whether H. citriformis strains may be developed as commercial products. In this review, we showed relevant information on isolation and bioassay strategies of H. citriformis to evaluate potential biocontrol strains under laboratory and field conditions in America. [ABSTRACT FROM AUTHOR]

The milk ring test is a detection assay for antibodies against Brucella in bovine milk. It has good sensitivity but tends to give false positive results. In this study, we standardized the application of the fluorescence polarization assay (FPA) for the detection of antibodies against B.melitensis in goat milk. We obtained negative serum and milk samples from healthy goat flocks in the northern zone of Nuevo León. Positive milk and negative, weak, and strong controls were obtained by mixing volumes of positive control serum with negative control milk. Milk samples were treated with citric acid, after which an FPA was performed. Results were then compared with the Rose Bengal test and the FPA in serum. Milk treatment allowed the quantification of antibodies in samples. Significant differences were found between the 2%, 4%, and 6% groups, compared with the control group (F3, 67 = 17.45, p < 0.0001) but not between the 2% and 4% groups (p = 0.0718). The cut-off value was 74.1 mP, with a sensitivity (Se) of 95% and a specificity (Sp) of 100%. Se and Sp values in field milk samples were 84% and 74.55%, respectively. Despite the FPA test on milk samples showed lower Se and Sp than the FPA test on serum samples, its cutoff may be adjusted. It may be recommended as a screening test in goat milk and become useful for the control and eradication of the disease. [ABSTRACT FROM AUTHOR]

Spodoptera frugiperda is a widely distributed insect pest that causes major economic losses in various crops, particularly maize. On the other hand, Beauveria bassiana is an entomopathogenic fungus that establishes symbiotic associations with many plants and contributes to tolerance against biotic and abiotic stresses. In the present work, in laboratory experiments, the effects of the B. bassiana strain GHA, in addition to a native strain (PTG4), delivered via seed treatment in maize seedlings, were evaluated on S. frugiperda growth, development, and mortality. We inoculated maize seeds with 1 × 106B. bassiana blastospores; then these seeds were germinated and grown to seedlings under growth chamber conditions. Third-instar S. frugiperda larvae were allowed to feed on B. bassiana-treated and -untreated (negative control) seedlings until reaching the sixth instar and transferred to an artificial diet until reaching adult stage. Results showed that larvae feeding on B. bassiana strain PTG4-treated plants prolonged their larval stage. Furthermore, feeding on plants treated with B. bassiana strains yielded fewer S. frugiperda male moths compared with feeding with the untreated control plants. Under field conditions, 1 × 106 (first trial) and 1 × 108 (second trial) of B. bassiana (GHA strain) blastospores were used for corn seed inoculation. In the first field trial, there were a higher number of larvae in the negative control plants compared to those in the plants treated with B. bassiana. No larvae were found in negative control and B. bassiana-treated plants in the second field trial. In conclusion, seed treatment with B. bassiana in maize reduced S. frugiperda infestation of maize plants in field trials. S. frugiperda development was also affected in laboratory trials. [ABSTRACT FROM AUTHOR]

Bioassay-guided fractionation of hexane extracts of Gymnosperma glutinosum (Asteraceae) leaves, collected in North Mexico, afforded the known compounds hentriacontane (1) and (+)-13S,14R,15-trihydroxy-ent-labd-7-ene (2), as well as the new ent-labdane diterpene (-)-13S,14R,15-trihydroxy-7-oxo-ent-labd-8(9)-ene (3). In addition, D-glycero-D-galactoheptitol (4) was isolated from the methanolic extract of this plant. Their structures were established on the basis of high-field 1D- and 2D NMR methods supported by HR-MS data. The cytotoxic activity was determined by using the in vitro L5178Y-R lymphoma murine model. Hentriacontane (1) and the new ent-labdane 3 showed weak cytotoxicity, whereas the ent-labdane 2 showed significant (p < 0.05) and concentration dependent cytotoxicity (up to 78%) against L5178Y-R cells at concentrations ranging from 7.8 to 250 μg/mL. [ABSTRACT FROM AUTHOR]