Fungal Load Research Articles

Chitosan nanoparticles encapsulating farnesol evaluated in vivo against Candida albicans.

Farnesol is a natural essential oil with antimicrobial properties. Complexation of farnesol in chitosan nanoparticles can be useful to improve its bioavailability and potentiate its antifungal capabilities such as inhibition of hyphal and biofilm formation. The aim of this study was to develop and characterize chitosan nanoparticles with farnesol (NF) and evaluate their toxicity and antifungal action on C. albicans in vivo. The NF were prepared by the ionic gelation method and showed physicochemical characteristics such as diameter less than 200 nm, monodisperse distribution, positive zeta potential, spherical morphology, and stability after 120 days of storage. In the evaluation of toxicity in Galleria mellonella, NF did not reduce the survival rate, indicating that there was no toxicity in vivo at the doses tested. In the assays with G. mellonella infected by C. albicans, the larvae treated with NF had a high survival rate after 48 h, with a significant reduction of the fungal load and inhibition of the formation of biofilms and hyphae. In the murine model of vulvovaginal candidiasis (VVC), histopathological analysis showed a reduction in inflammatory parameters, fungal burden, and hyphal inhibition in mice treated with NF. The produced nanoparticles can be a promising alternative to inhibit C. albicans infection.

Ethyl caffeate combined with fluconazole exhibits efficacy against azole-resistant oropharyngeal candidiasis via the EFGR/JNK/c-JUN signaling pathway.

Ethyl caffeate (EC) is a phenylpropanoid compound derived from Elephantopus scaber. In our previous work, EC was investigated to have a strong synergistic antifungal effect against azole-resistant strains of Candida albicans when combined with fluconazole (FLU). However, the protective effect and mechanism of EC+FLU on oropharyngeal candidiasis (OPC) caused by drug-resistant strains of C. albicans has not been investigated. This study aimed to investigate the protective effect and mechanism of EC combined with FLU against C. albicans-resistant strains that lead to OPC. An OPC mouse model revealed that EC+FLU treatment reduced fungal load and massive hyphal invasion of tongue tissues, and ameliorated the integrity of the tongue mucosa. PAS staining results showed more structural integrity of the tongue tissues and reduced inflammatory cell infiltration after EC+FLU treatment. Phosphorylation of EGFR and other proteins in the EFGR/JNK/c-JUN signaling pathway was significantly downregulated by EC+FLU. EGFR and S100A9 mRNA expression was also reduced. The above results were verified in FaDu cells. ELISA results showed that the concentration of inflammatory factors in the cell supernatant was significantly reduced after EC combined with FLU treatment. Molecular docking revealed that EC exhibited high binding energy to EGFR. In conclusion, EC enhances the susceptibility of azole-resistant C. albicans to FLU, and the underlying mechanism is related to the inhibition of the EGFR/JNK/c-JUN signaling pathway. This result suggests that EC has potential to be developed as an antifungal sensitizer to treat OPC caused by azole-resistant C. albicans.

Sporothrix brasiliensis Infection Modulates Antimicrobial Peptides and Stress Management Gene Expression in the Invertebrate Biomodel Galleria mellonella.

Sporothrix brasiliensis is the most pathogenic species, responsible for the Brazilian cat-transmitted sporotrichosis hyperendemic. In this scenario, an investigation of the pathogen-host interaction can provide relevant information for future treatment strategies. To this end, the invertebrate Galleria mellonella has proven to be a suitable alternative for evaluating the virulence of pathogenic fungi, since the insect immune system is similar to the mammalian innate immune response. The aim of this work was to investigate phenotypic and molecular aspects of the immune response of G. mellonella throughout the S. brasiliensis infection. Hemocyte density and the evolution of the fungal load were evaluated. In parallel, RT-qPCR expression analysis of genes encoding antimicrobial peptides (Gallerimycin and Galiomycin) and stress management genes (C7 Contig 15362 and C8 Contig 19101) was conducted. The fungal load and hemocyte densities increased simultaneously and proportionally to the deleterious morphological events and larvae mortality. Gallerimycin, C7 Contig 15362 and C8 Contig 19101 genes were positively regulated (p < 0.05) at distinct moments of S. brasiliensis infection, characterizing a time-dependent and alternately modulated profile. Galiomycin gene expression remained unchanged. Our results contribute to the future proposal of potential alternative pathways for treating and consequently controlling S. brasiliensis zoonosis, a major public health issue in Latin America.

Non-touch treatment to solve the persistent contamination of culture plates with Aspergillus niger in a clinical microbiology laboratory of Spain.

A pseudo-outbreak of Aspergillus caused by false positive cultures can have a high sanitary impact. We determined the effectiveness (fungal load elimination) of a non-touch disinfection system, vs conventional disinfection methods, to solve steady contamination of culture plates with Aspergillus niger at a clinical microbiology laboratory. Routine cleaning-disinfection (RCD), intensive cleaning-disinfection (ICD), and terminal airborne disinfection (TAD) were employed in stages. Air sampling was carried out before and after each procedure. The effectiveness of TAD on contact surfaces was tested by surface sampling. After RCD, ICD, and TAD, there was a mean decrease of 5.4 (95% CI = 1.8-9.0), 19.2 (95% CI = 12.4-26.0), and 4.4 (95% CI = 2.5-6.3) CFU per tested area, and 46.2%, 21.7%, and 95.5% of contaminated areas became sterile, respectively. There was a mean decrease of 30.6CFU per tested surface (p < 0.0007) and 50% of tested surfaces became sterile. Global effectiveness of RCD, ICD, and TAD was 68.8% (95% CI = 68.5-69.1), 82.2% (95% CI = 82.1-82.3), and 99.0% (95% CI = 98.8-99.2), respectively. The effectiveness was higher with TAD (4.1CFU/cm2 less than with ICD; p = 0.0290). No further contamination has occurred since then. When construction and renovation activities are discarded and RCD and ICD practices are insufficient, non-touch disinfection remove residual dust contamination and avoid recurrence.

Curcuminoid-Mediated Antimicrobial Photodynamic Therapy on a Murine Model of Oral Candidiasis.

Antimicrobial Photodynamic Therapy (aPDT) has been extensively investigated in vitro, and preclinical animal models of infections are suitable for evaluating alternative treatments prior to clinical trials. This study describes the efficacy of aPDT in a murine model of oral candidiasis. Forty mice were immunosuppressed with subcutaneous injections of prednisolone, and their tongues were inoculated using an oral swab previously soaked in a C. albicans cell suspension. Tetracycline was administered via drinking water during the course of the experiment. Five days after fungal inoculation, mice were randomly distributed into eight groups; a ninth group of untreated uninfected mice was included as a negative control (n = 5). Three concentrations (20 µM, 40 µM, and 80 µM) of a mixture of curcuminoids were tested with a blue LED light (89.2 mW/cm2; ~455 nm) and without light (C+L+ and C+L- groups, respectively). Light alone (C-L+), no treatment (C-L-), and animals without infection were evaluated as controls. Data were analyzed using Welch's ANOVA and Games-Howell tests (α = 0.05). Oral candidiasis was established in all infected animals and visualized macroscopically through the presence of characteristic white patches or pseudomembranes on the dorsum of the tongues. Histopathological sections confirmed a large presence of yeast and filaments limited to the keratinized layer of the epithelium in the C-L- group, and the presence of fungal cells was visually decreased in the images obtained from mice subjected to aPDT with either 40 µM or 80 µM curcuminoids. aPDT mediated by 80 µM curcuminoids promoted a 2.47 log10 reduction in colony counts in comparison to those in the C-L- group (p = 0.008). All other groups showed no statistically significant reduction in the number of colonies, including photosensitizer (C+L-) or light alone (C-L+) groups. Curcuminoid-mediated aPDT reduced the fungal load from the tongues of mice.

Airborne Culturable Fungi in Primary Schools

The aim of this current research was to investigate airborne fungi in indoor environments at primary schools. The study also examined the correlation between these fungi and building characteristics, as well as some geographical and meteorological parameters. To achieve this aim, a passive sampling method was used. In this study, 148 samples were collected from the indoor environments of 24 schools located in Qom, Iran. To collect the samples, passive sampling was performed using Petri plates containing Sabouraud dextrose agar (SDA). The characteristics of school buildings were evaluated by checklist. The mean ± SD fungal load of indoor air in selected schools was found to be 10.1 ± 14.0 colonies (CFU/dm2/ h). According to the IMA standard, the majority of the classes (71.7%) were in very good condition. The dominant species were as follows: Aspergillus niger, Aspergillus candidus, and Aspergillus flavus. Furthermore, the fungal load of girls’ schools was significantly higher than that of the boys' schools (p &lt; .05). The correlation analysis using the Pearson test showed that there was a direct correlation between the mean fungal load of classrooms and the number of students (p &lt; .01). The highest concentration of fungi was found on the ground floor and in poor ventilation conditions (p &lt; .05). During the study, it was found that schools located in the western part of Qom, Iran, had a higher concentration of fungi. This can be attributed to their exposure to the prevailing winds and the penetration of outdoor dusty air into indoor environments. The large number of students in each class and the inappropriate ventilation, which are the causes of airborne culturable fungi of these classes, call for the need for proper operation of school buildings.

Differential skin immune responses in mice intradermally infected with Candida auris and Candida albicans.

Candida auris is a globally emerging fungal pathogen that transmits among individuals in hospitals and nursing home residents. Unlike other Candida species, C. auris predominantly colonizes and persists in skin tissue, resulting in outbreaks of nosocomial infections. Understanding the factors that regulate C. auris skin colonization is critical to develop novel preventive and therapeutic approaches against this emerging pathogen. We established a model of intradermal C. auris inoculation in mice and found that mice infected with C. auris elicit less potent innate and adaptive immune responses in the infected skin compared to C. albicans. These findings help explain the clinical observation of persistent C. auris colonization in skin tissue.

Thymoquinone attenuates inflammation in C. Albicans keratitis by activating Nrf2/HO-1 signaling pathway and reducing fungal load

PurposeThis study aims to investigate the anti-inflammatory and antifungal properties of thymoquinone (TQ) and elucidate its mechanism of action in the context of C. albicans keratitis. MethodsVarious methods were employed to identify a safe and effective concentration of TQ with antifungal properties, including the determination of the minimum inhibitory concentration (MIC), the cell counting kit-8 (CCK-8) test, and the Draize experiment. The severity of fungal keratitis (FK) was assessed through clinical ratings and slit-lamp imaging. Fungus burden was determined using plate counting and periodic acid Schiff (PAS) staining. Neutrophil infiltration and activity were investigated through immunofluorescence staining (IFS), myeloperoxidase (MPO) analysis, and hematoxylin and eosin (HE) staining. To explore the anti-inflammatory effects of TQ and its mechanism of action, we employed RT-PCR, ELISA, and western blot techniques. ResultsTQ effectively controlled fungal growth at a concentration of 50 µg/mL while preserving the integrity of mouse corneas. Human corneal epithelial cells (HCECs) remained unaffected by TQ at concentrations ≤ 3.75 µg/mL. Treatment with TQ led to significant improvements in clinical scores, fungal burden, neutrophil infiltration, and the expression of inflammatory factors compared to the DMSO group. Moreover, TQ demonstrated the ability to reduce the levels of inflammatory factors in HCECs stimulated by C. albicans. Additionally, TQ enhanced the expressions of Nrf2 and HO-1 in mouse corneas. The downregulation of cytokines induced by TQ was reversed upon pretreatment with inhibitors of Nrf2 or HO-1. ConclusionTQ exhibits a protective effect in the context of C. albicans keratitis through multiple mechanisms, including inhibition of C. albicans growth, reduction of neutrophil recruitment, activation of the Nrf2/HO-1 pathway, and limitation of the expression of pro-inflammatory factors.

Dynamic Reconfiguration of Switchgrass Proteomes in Response to Rust (Puccinia novopanici) Infection.

Switchgrass (Panicum virgatum L.) can be infected by the rust pathogen (Puccinia novopanici) and results in lowering biomass yields and quality. Label-free quantitative proteomics was conducted on leaf extracts harvested from non-infected and infected plants from a susceptible cultivar (Summer) at 7, 11, and 18 days after inoculation (DAI) to follow the progression of disease and evaluate any plant compensatory mechanisms to infection. Some pustules were evident at 7 DAI, and their numbers increased with time. However, fungal DNA loads did not appreciably change over the course of this experiment in the infected plants. In total, 3830 proteins were identified at 1% false discovery rate, with 3632 mapped to the switchgrass proteome and 198 proteins mapped to different Puccinia proteomes. Across all comparisons, 1825 differentially accumulated switchgrass proteins were identified and subjected to a STRING analysis using Arabidopsis (A. thaliana L.) orthologs to deduce switchgrass cellular pathways impacted by rust infection. Proteins associated with plastid functions and primary metabolism were diminished in infected Summer plants at all harvest dates, whereas proteins associated with immunity, chaperone functions, and phenylpropanoid biosynthesis were significantly enriched. At 18 DAI, 1105 and 151 proteins were significantly enriched or diminished, respectively. Many of the enriched proteins were associated with mitigation of cellular stress and defense.

Quercetin protects against Aspergillus fumigatus keratitis by reducing fungal load and inhibiting TLR-4 induced inflammatory response

PurposeTo investigate the antifungal and anti-inflammatory effects of quercetin in Aspergillus fumigatus (A. fumigatus) keratitis. MethodsDraize eye test was performed in mice to evaluate the toxicity of quercetin, and the antifungal effects on A. fumigatus were assessed via scanning electron microscopy (SEM), propidium iodide uptake, and adherence assay. In fungal keratitis (FK) mouse models, immunostaining was performed for investigating toll-like receptor 4 (TLR-4) expression and macrophage infiltration. Real-time PCR, ELISA, and Western blot were used to evaluate the expression of pro-inflammatory factors IL-1β, TNF-α, and IL-6 in infected RAW264.7 cells. Cells were also treated with TLR-4 siRNA or agonist CRX-527 to investigate mechanisms underlying the anti-inflammatory activity of quercetin. ResultsQuercetin at 32 μM was non-toxic to corneal epithelial and significantly inhibited A. fumigatus growth and adhesion, and also altered the structure and reduced the number of mycelia. Quercetin significantly reduced macrophage infiltration in the mouse cornea, and attenuated the expression of TLR-4 in the corneal epithelium and stroma of mice with keratitis caused by A. fumigatus. In RAW264.7 cells infected by A. fumigatus, quercetin downregulated TLR-4 along with pro-inflammatory factors IL-1β, TNF-α, and IL-6. RAW cells with TLR-4 knockdown had reduced expression of factors after A. fumigatus infection, which was decreased even further with quercetin treatment. In contrast, cells with CRX-527 had elevated inflammatory factors compared to control, which was significantly attenuated in the presence of quercetin. ConclusionQuercetin plays a protective role in mouse A. fumigatus keratitis by inhibiting fungal load, disrupting hyphae structure, macrophage infiltration, and suppressing inflammation response in macrophages via TLR-4 mediated signaling pathway.

Construction of a murine model of latent infection and reactivation induced by Talaromyces marneffei

ObjectiveTo establish a murine model of Talaromyces marneffei (T. marneffei) latent infection and reactivation, providing a foundation for exploring the molecular mechanisms underlying disease relapse. MethodsBALB/c mice were tail vein injected with T. marneffei at 0 days post-infection (dpi) and treated with cyclophosphamide (CTX) intraperitoneally every four days, starting from 21 dpi or 42 dpi. Mice were observed for body weight changes, liver and spleen indices, histological characteristics of liver and spleen, fungal load detection in liver and spleen, and Mp1p qualitation in liver and spleen to assess T. marneffei infection severity. ResultsT. marneffei-infected mice exhibited a trend of initial weight loss followed by recovery and a subsequent decrease in weight after CTX injection throughout the observation period. Liver and spleen indices, as well as tissue damage, significantly increased during infection but later returned to normal levels, with a gradual rise observed after immunosuppression. Fungal load analysis revealed positive T. marneffei cultures in the liver and spleen at 7 dpi and 14 dpi, followed by negative T. marneffei cultures from 21 dpi until day 21 post-immunosuppression (42 dpi or 63 dpi); however, the spleen remained T. marneffei-cultured negative, consistent with the trend observed in Mp1p detection results. ConclusionA latent infection and reactivation model of T. marneffei in mice was successfully established, with the liver likely serving as a key site for latent T. marneffei.

Harmonizing Drying Time, Layer Thickness, and Drier Zones for Drying Kinetics: Quality and Safety of Solar Tunnel-Dried Wet-Processed Parchment Coffee (Coffea arabica L.).

Tunnel solar dryer is the recently used drying method for better quality and safety of parchment coffee. However, the higher variation of drying temperature and RH along the long tunnel solar dryer results in a heterogeneous environment in the tunnel, which could make parchment coffee dried at different times or with different moisture contents. This study is aimed at investigating the effect of solar tunnel dryer zones at different zones of the dryer, divided into three zones from the inlet to the exit side of the drier and drying layer thicknesses on the drying time, drying kinetics, physicochemical, sensory, and fungal growth loads of parchment coffee. Furthermore, seven mathematical models were evaluated to select the best-fitting model for a specific zone to predict drying time. Results showed that dryer zones significantly (p < 0.05) interacted with layer thickness for most of the measured parameters except titratable acidity and sensory properties. The dryer zone, coupled with the reduction in drying layer thickness, caused an increase in effective diffusivity and moisture removal rate and reduced drying time. The drying time to reach constant moisture content varied from 14 to 17 hours. Overall raw bean, cup, and total quality varied from 36.3 to 37, 48 to 51, and 84.3 to 87.3%, respectively. Values for physicochemical parameters ranged from 5.3 to 6.9 (pH), 2.1 to 2.6% (titratable acidity), 2.3 to 4.3°Brix TSS, 10.9 to 15.2% (ether extract), 39.2 to 53.5GAE/g (total phenolic content), and 38.5 to 59.2 (DPPH scavenging capacity). The fungal infection percentage at the end of drying varied from 4 to 93.3%, which could be associated with potential mycotoxin formation if recommended conditions were not maintained. In general, for better quality, similar drying times, and a lesser fungal load, it is recommended to use 4, 5, and 6 cm layer thickness in zones one, two, and three, respectively. The drying kinetics of parchment coffee in different dryer zones with different drying layer thicknesses showed variation. Zone one at 2 and 4 cm layer thicknesses is best described by the Verma model. Four- and six-centimetre layer thicknesses in zones 2 and 3 are best described by the modified Midilli model.

Increased susceptibility to encephalitozoonosis associated with mixed Th1/Th2 profile and M1/M2 profile in mice immunosuppressed with cyclophosphamide

Encephalitozoon cuniculi is a unicellular, spore-forming, obligate intracellular eukaryote belonging to the phylum Microsporidia. It is known to infect mainly immunocompromised and immunocompetent mammals, including humans. The parasite–host relationship has been evaluated using both in vitro cell culturing and animal models. For example, Balb/c and C57BL/6 mouse strains have been used interchangeably, although the latter has been considered more susceptible due to the higher fungal load observed after infection. In the present study, we identified the characteristics of the immune response of C57BL/6 mice treated or not with the immunosuppressant cyclophosphamide (Cy) and challenged with E. cuniculi by intraperitoneal route. After 14 days of infection, serum was collected to analyze Th1, Th2, and Th17 cytokine levels. In addition, peritoneal washes were performed, and the spleen sample was collected for immune cell phenotyping, whereas liver, spleen, kidney, lung, intestine, and central nervous system (CNS) samples were collected for histopathological analysis. Although infected mice displayed a reduced absolute number of macrophages, they showed an M1 profile, an elevated number of CD4+T, CD8+T, B-1, and B-2 lymphocytes, with a predominance of Th1 inflammatory cytokines (interferon [IFN]-γ, tumor necrosis factor [TNF]-α, and interleukin [IL]-2) and Th17. Furthermore, Cy-Infected mice showed a reduced absolute number of macrophages with an M1 profile but a reduced number of CD4+T, CD8+T, B-1, and B-2 lymphocytes, with a predominance of Th1 inflammatory cytokines (IFN-γ, TNF-α, and IL-2) and Th2 (IL-4). This group displayed a higher fungal burden as well and developed more severe encephalitozoonosis, which was associated with a reduced number of T and B lymphocytes and a mixed profile of Th1 and Th2 cytokines.

MYCOFLORA OF FERMENTED LOCUST BEANS (Parkia biglobosa) SEED, “IRU” SOLD IN LOKOJA MARKETS, KOGI STATE, NIGERIA

Fermented locust beans are a popular nutritious food seasoning that are prone to microbial contamination due to unhygienic handling and improper storage. This research aimed to evaluate the mycological quality of locust beans sold in Lokoja markets, Kogi State, Nigeria. Thirty (30) samples of locust beans were collected from five different markets in Lokoja, analyzed for inherent fungi which were identified using standard mycological techniques. The results indicated variations in the mean fungal loads across the sampled markets. Old market had the highest mean fungal load (4.33 × 105 CFU/g), followed by Adankolo market (3.67 × 105 CFU/g) and Kpata market (3.0 × 105 CFU/g) while Lokongoma market had the lowest mean fungal load (2.0 × 105 CFU/g). The mean fungal load of the fermented locust beans from Old market was significantly different (p&gt;0.05). A total of 55 fungal isolates that include: Aspergillus niger (29.1 %), Rhizopus arrhizus (25.5 %), Fusarium cladosporium (18.2 %), Fusarium cummunis (12.7 %), Aspergillus nudilans (9.1%), Aspergillus tamari (7.3 %) and Aspergillus terreus (1.8%) were obtained from the samples. Aspergillus niger, F. communis, F. clamadosporium and R. arrhizus were present in all the sampled markets, suggesting they are the predominant fermenter of locust beans. The presence of mycotoxigenic fungi in the locust beans seeds poses a significant public health risk. Therefore, it is important to ensure that locust beans are processed and stored under hygienic conditions at all times to safeguard consumer health.

The respiratory tract microbiome, the pathogen load, and clinical interventions define severity of bacterial pneumonia

Bacterial pneumonia is a considerable problem worldwide. Here, we follow the inter-kingdom respiratory tract microbiome (RTM) of a unique cohort of 38 hospitalized patients (n= 97 samples) with pneumonia caused by Legionella pneumophila. The RTM composition is characterized by diversity drops early in hospitalization and ecological species replacement. RTMs with the highest bacterial and fungal loads show low diversity and pathogen enrichment, suggesting high biomass as a biomarker for secondary and/or co-infections. The RTM structure is defined by a "commensal" cluster associated with a healthy RTM and a "pathogen" enriched one, suggesting that the cluster equilibrium drives the microbiome to recovery or dysbiosis. Legionella biomass correlates with disease severity and co-morbidities, while clinical interventions influence the RTM dynamics. Fungi, archaea, and protozoa seem to contribute to progress of pneumonia. Thus, the interplay of the RTM equilibrium, the pathogen load dynamics, and clinical interventions play a critical role in patient recovery.

Development of Echinocandin Resistance in Candida haemulonii: An Emergent, Widespread, and Opportunistic Fungal Pathogen.

Echinocandins, used for the prevention and treatment of invasive fungal infections, have led to a rise in breakthrough infections caused by resistant Candida species. Among these species, those belonging to the Candida haemulonii complex are rare multidrug-resistant (MDR) yeasts that are frequently misidentified but have emerged as significant healthcare-associated pathogens causing invasive infections. The objectives of this study were to investigate the evolutionary pathways of echinocandin resistance in C. haemulonii by identifying mutations in the FKS1 gene and evaluating the impact of resistance on fitness. After subjecting a MDR clinical isolate of C. haemulonii (named Ch4) to direct selection using increasing caspofungin concentrations, we successfully obtained an isolate (designated Ch4'r) that exhibited a high level of resistance, with MIC values exceeding 16 mg/L for all tested echinocandin drugs (caspofungin, micafungin, and anidulafungin). Sequence analysis revealed a specific mutation in the resistant Ch4'r strain, leading to an arginine-histidine amino acid substitution (R1354H), occurring at the G4061A position of the HS2 region of the FKS1 gene. Compared to the wild-type strain, Ch4'r exhibited significantly reduced growth proliferation, biofilm formation capability, and phagocytosis ratio, indicating a decrease in fitness. Transmission electron microscopy analysis revealed alterations in cell wall components, with a notable increase in cell wall thickness. The resistant strain also exhibited higher amounts (2.5-fold) of chitin, a cell wall-located molecule, compared to the wild-type strain. Furthermore, the resistant strain demonstrated attenuated virulence in the Galleria mellonella larval model. The evolved strain Ch4'r maintained its resistance profile in vivo since the treatment with either caspofungin or micafungin did not improve larval survival or reduce the fungal load. Taken together, our findings suggest that the acquisition of pan-echinocandin resistance occurred rapidly after drug exposure and was associated with a significant fitness cost in C. haemulonii. This is particularly concerning as echinocandins are often the first-line treatment option for MDR Candida species.

Correlation between Aspergillus flavus fungal biomass and aflatoxin contamination in harvested maize: insights from Kenya and Tanzania

Controlling the occurrence of aflatoxins in foods must be accompanied by managing the fungi responsible for their production. The abundance and diversity of aflatoxin-producing Aspergillus flavus are responsible for the accumulation of these toxins in crops, posing a persistent threat to public health and the economy in tropical developing countries. A study was conducted to investigate the occurrence and level of A. flavus and relate them to aflatoxin levels in maize in Kenya and Tanzania. A total of 786 maize samples were collected during harvesting in selected areas of the two countries for analysis. The fungal abundance in the samples was measured as the amount of fungal DNA relative to maize DNA. This was accomplished by quantifying the fungal DNA using qPCR, targeting the internal transcribed spacer (ITS) gene, while the maize DNA was quantified through the alpha-tubulin gene, the two genes known to be conserved. Aflatoxins were quantified using ultra-high performance liquid chromatography, coupled with ultra-high sensitivity, ultra-fast triple quadrupole tandem-mass spectrophotometer. A. flavus was detected in 88.5% of the 786 tested samples, and the average fungal load for these samples (expressed as the log host/pathogen ratio) was 5.53. Aflatoxin occurrence was positive in 31.9% of the samples, with an average level of 2.3 ± 0.643 ppb. The study established a positive relationship between the occurrence and level of aflatoxin B1 and the presence and biomass of A. flavus, which was statistically proven. These findings emphasize the need to place substantial attention on preharvest control of A. flavus in cereal fields as an effort to control the accumulation of aflatoxin B1 in foods.

Environmental factors influencing indoor airborne fungi in students dormitory - a case study in Nakhon Sithammarat, Thailand.

Indoor airborne fungi are a significant health concern that can cause respiratory symptoms and other health problems. Indoor fungi are influenced by various factors such as meteorological conditions and dwelling characteristics. This study aims to evaluate the association between indoor airborne fungi and environmental factors in a student dormitory in southern Thailand. The study was conducted at Walailak University in southern Thailand from September toDecember 2020. Air samples were collected from rooms in thirteen dormitories, and the fungal load was determined using the passive air sampling method. The study also measured meteorological parameters and gathered data on occupant behaviors and exposure-related symptoms through a self-administered questionnaire. In a total of 135 student rooms, the average concentration (mean ± SD) of indoor airborne fungi was 409.72±176.22 CFU/m3, which showed the highest concentration on the first floor. For meteorological parameters, the averages of RH (%), temperature (°C), and CO2 (ppm) were 70.99±2.37, 31.11±0.56 and 413.29±76.72, respectively. The abundance of indoor airborne fungi was positively associated with an increase in RH (β=0.267, 95% CI: 5.288, 34.401) and building height (β=0.269, 95% CI: 16.283, 105.873), with values of 19.845 and 61.078, respectively. Conversely, temperature exhibited a negative effect on indoor airborne fungi (-92.224, β=-0.292, 95% CI: -150.052, -34.396). The findings highlight the influence of RH, temperature and building height on indoor airborne fungi in the student dormitory. Therefore, effective management strategies are necessary to improve indoor air quality and reduce associated health risks in student dormitories.

Use of X-Irradiations in Reducing the Waste of Aflatoxin-Contaminated Pistachios and Evaluation of the Physicochemical Properties of the Irradiated Product.

This study investigates the effects of electron beam irradiation (0, 1, 2, 4, and 6 kGy) on Aspergillus flavus, aflatoxin B1 (AFB1), and the physicochemical properties of pistachios. The findings suggested that e-beam significantly reduced the spore population of A. flavus and the concentration of AFB1 at doses of 4 and 6 kGy. Three AFB1 degradation products were detected via LC-MS analysis and their structures were presented. Total phenolic content was improved at a dose of 2 kGy, while antioxidant activity was decreased in all treatments in both DPPH and ABTS assays. The chlorophyll and carotenoid content declined and the color indices changed, leading to a darker color. E-beam at a dose of 2 kGy raised the soluble protein levels and changed the intensity and pattern of protein bands. Irradiation doses of up to 6 kGy enhanced the content of malondialdehyde and total saturated fatty acids while leading to a decline in unsaturated fatty acids. The quality features were adversely affected at doses > 4 kGy. The findings suggest that as an alternative method, e-beam at doses ≥ 2 kGy can effectively decrease fungal load and aflatoxin B1 contamination, and e-beam application at doses ≤ 2 kGy can maintain the physicochemical attributes of pistachios to an acceptable extent.

Ascending renal infection following experimental candiduria by Candida tropicalis in immunocompromised mice

The present study evaluated renal infection resulting from the implantation of C. tropicalis in the bladder of immunosuppressed mice. Yeasts were implanted in two manners: planktonic and via preformed biofilm on a small catheter fragment (SCF). Renal histopathology and cultures was performed 72 and 144 h after cystotomy was carried out in mice from three groups: group I contained non-contaminated mice implanted with a sterile SCF; group II mice received a sterile SCF plus a yeast suspension containing 1 × 107 yeasts/mL in a planktonic form; group III mice were implanted with a SCF containing preformed C. tropicalis biofilm. Viable yeasts were found in the kidneys of mice from both groups II and III. However, after 72 h the planktonic cells (group II) invaded more quickly than the sessile cells (group III). Over a longer period (144 h), group III exhibited a more invasive infection (50% of the animals presented renal infection and the renal fungal load was 3.2 log10 CFU/g tissue) than in group II, where yeasts were not found. C. tropicalis introduced into the bladder in two ways (in planktonic or biofilm form) were able to reach the kidney and establish a renal fungal infection, causing interstitial disorders. The data of the present study therefore support the hypothesis of an ascending pathway for renal infections by C. tropicalis. Furthermore, the biofilm resulted in a greater and progressive risk of renal infection, attributed to the slow detachment of the yeasts.