Effect of Christmas Melon (Laganaria Breviflorus) extract on toxigenic Mycoflora Isolated from Stored Unpolished Rice sold in major Markets in Abeokuta, Nigeria

Study on toxigenic mycoflora and potential mitigation effect of Christmas Melon (Laganaria Breviflorus) extract in unpolished rice sold in Abeokuta Ogun state of Nigeria was carried out. Unpolished rice gotten from markets in Abeokuta were aseptically transported to the laboratory, serial dilution to reduce the fungal load was carried out and were plated on Potato Dextrose Agar (PDA) and Methyl Red Dessicated Coconut Agar (MRDCA) respectively. Microscopy, macroscopy, toxigenicity test and inhibition studies with the peeled and unpeeled fruit of Laganaria breviflorus fermented for seven days was carried out. Results reveal the predominance of Aspergillus as the major genera, specifically, A. niger, A.flavus, A. parasiticus, A. fumigatus, A. terreus, A. nidulans. Other fungi genera isolated include Penicillium, F`usarium, Mucor, Alternaria and Rhizopus . Of the 11 fungi genera isolated, 9 were toxigenic of which the zones of inhibition of unpeeled whole fruit extract of Laganaria breviflorus range from (3 - 28mm) where A. nidulans shows the highest susceptibility to the whole fruit extract of Laganaria breviflorus while the zone of inhibition of peeled fruit extract of Laganaria breviflorus ranges from (3 - 22mm) where A. parasiticus, Fusarium specie and P.chrysogenum showed the highest susceptibility . As the day progresses the zone of inhibition becomes wider. Unpeeled LB extract exhibited more zones of inhibition than the peeled LB extract. Laganaria breviflorus fruit extracts in the study demonstrates a potential in reducing toxigenic fungi, consequently a means to reducing mycotoxins in staple foods in Nigeria.

Interventional Effect of Nanosilver Paint on Fungal Load of Indoor Air in a Hospital Ward

Hospital ward environments contain various types of microorganisms, in which fungal agents are one of the main contaminants that may cause hospital-acquired infections. Regarding this, the aim of the present study was to evaluate the effect of nanosilver paint on reducing fungal contaminants of indoor air in an educational, research, and treatment center. Two rooms in the hematology ward were selected. One room was painted using usual paint (control room) and the other room was painted with paint containing nanosilver particles (experimental room). One hundred and twelve samples were collected using active (Anderson BioSampler) and passive (settle plate or open plate) air sampling techniques. The samples were incubated for 3–7 days at 35°C, and the positive fungal cultures were examined according to morphological and microscopic characteristics. Following active sampling, the mean and standard deviation of the number of colony-forming units (CFU/m3) of fungi colonies in the experimental and control rooms were 29.21 ± 17.99 and 22.50 ± 10.02 before intervention and 13.79 ± 6.20 and 31.07 ± 21.1 after intervention, respectively. Following passive sampling, the number of CFU/plate in the experimental and control rooms was 6 and 0 before and 1and 1 after intervention, respectively. The use of the nanosilver paint was effective in reducing air fungal contamination. Moreover, the active sampling method was more sensitive to measuring the concentration changes for fungal bioaerosols.

Fungal Population and Physicochemical Characteristics of Abattoir-Impacted Soil in Iwofe, Rivers State

Aim: To determine the fungal population and physicochemistry of abattoir impacted soil in Iwofe, Rivers State. Study Design: This study focused on Abattoir impacted soil. Statistical analysis of data and interpretation was carried out. Place and Duration of Study: Abattoir impacted soil was collected from three points in an abattoir located in Iwofe, Rivers State while the unpolluted soil which served as control was collected from the Rivers State University, Port Harcourt in January, 2021. Methodology: Standard microbiological techniques were used: the fungal population was determined by inoculating aliquots of an appropriate dilution resulting from a ten-fold serial dilution on prepared Sabouraud dextrose agar plates in duplicates. Plates were later incubated for 3-5 days after which colonies were enumerated and used in obtaining the fungal population in the soil samples while distinct colonies were subcultured for macroscopic and microscopic identification of fungi. The physicochemical parameters and heavy metals were analyzed using standard methods. Results: Fungal load in the control and abattoir impacted soil were 1.09×105 and 3.9×104 CFU/g, respectively. The fungal load of the control soil was significantly higher (P˂0.05) than the abattoir impacted soil. The fungal isolates identified in the abattoir impacted soil were Microsporium sp, Aspergillus niger and Candida sp while Aspergillus niger, Aspergillus flavus, Fusarium sp, Penicillium sp, Mucor sp and Rhizopus sp were identified from the control soil. The pH, temperature, nitrate and phosphate of the abattoir soil were 6.7, 28.33℃, 27.83(mgKg-1) and 1055(mgKg-1), respectively. The concentrations of Cadmium, Iron and Lead in the abattoir Impacted soil and control soil were 0.81, 563.35 and 7.12 mgKg-1, 0.51, 582.0 and 3.18 mgKg-1, respectively. The physico chemistry and heavy metals in the abattoir soil were within acceptable limits. Discussion and Conclusion: The findings from this study showed that heavy metals in abattoir impacted soil had an impact in the fungal population which led to the isolation of only three fungal isolates belonging to Microsporium sp, Candida sp and Aspergillus niger. More so, despite the presence of heavy metals in the abattoir impacted soil, the metals were all within permissible limits. Thus, the abattoir impacted soil was not heavily polluted.

Tryptophan metabolism and bacterial commensals prevent fungal dysbiosis in Arabidopsis roots

In nature, roots of healthy plants are colonized by multikingdom microbial communities that include bacteria, fungi, and oomycetes. A key question is how plants control the assembly of these diverse microbes in roots to maintain host–microbe homeostasis and health. Using microbiota reconstitution experiments with a set of immunocompromised Arabidopsis thaliana mutants and a multikingdom synthetic microbial community (SynCom) representative of the natural A. thaliana root microbiota, we observed that microbiota-mediated plant growth promotion was abolished in most of the tested immunocompromised mutants. Notably, more than 40% of between-genotype variation in these microbiota-induced growth differences was explained by fungal but not bacterial or oomycete load in roots. Extensive fungal overgrowth in roots and altered plant growth was evident at both vegetative and reproductive stages for a mutant impaired in the production of tryptophan-derived, specialized metabolites (cyp79b2/b3). Microbiota manipulation experiments with single- and multikingdom microbial SynComs further demonstrated that 1) the presence of fungi in the multikingdom SynCom was the direct cause of the dysbiotic phenotype in the cyp79b2/b3 mutant and 2) bacterial commensals and host tryptophan metabolism are both necessary to control fungal load, thereby promoting A. thaliana growth and survival. Our results indicate that protective activities of bacterial root commensals are as critical as the host tryptophan metabolic pathway in preventing fungal dysbiosis in the A. thaliana root endosphere.

NLRP3 Inflammasome Contributes to Host Defense Against Talaromyces marneffei Infection

Talaromyce marneffei is an important thermally dimorphic pathogen causing disseminated mycoses in immunocompromised individuals in southeast Asia. Previous studies have suggested that NLRP3 inflammasome plays a critical role in antifungal immunity. However, the mechanism underlying the role of NLRP3 inflammasome activation in host defense against T. marneffei remains unclear. We show that T. marneffei yeasts but not conidia induce potent IL-1β production. The IL-1β response to T. marneffei yeasts is differently regulated in different cell types; T. marneffei yeasts alone are able to induce IL-1β production in human PBMCs and monocytes, whereas LPS priming is essential for IL-1β response to yeasts. We also find that Dectin-1/Syk signaling pathway mediates pro-IL-1β production, and NLRP3-ASC-caspase-1 inflammasome is assembled to trigger the processing of pro-IL-1β into IL-1β. In vivo, mice deficient in NLRP3 or caspase-1 exhibit higher mortality rate and fungal load compared to wild-type mice after systemic T. marneffei infection, which correlates with the diminished recruitment of CD4 T cells into granulomas in knockout mice. Thus, our study first demonstrates that NLRP3 inflammasome contributes to host defense against T. marneffei infection.

Impacts of fungal entomopathogens on survival and immune responses of Aedes albopictus and Culex pipiens mosquitoes in the context of native Wolbachia infections

Microbial control of mosquitoes via the use of symbiotic or pathogenic microbes, such as Wolbachia and entomopathogenic fungi, are promising alternatives to synthetic insecticides to tackle the rapid increase in insecticide resistance and vector-borne disease outbreaks. This study evaluated the susceptibility and host responses of two important mosquito vectors, Ae. albopictus and Cx. pipiens, that naturally carry Wolbachia, to infections by entomopathogenic fungi. Our study indicated that while Wolbachia presence did not provide a protective advantage against entomopathogenic fungal infection, it nevertheless influenced the bacterial / fungal load and the expression of select anti-microbial effectors and phenoloxidase cascade genes in mosquitoes. Furthermore, although host responses from Ae. albopictus and Cx. pipiens were mostly similar, we observed contrasting phenotypes with regards to susceptibility and immune responses to fungal entomopathogenic infection in these two mosquitoes. This study provides new insights into the intricate multipartite interaction between the mosquito host, its native symbiont and pathogenic microbes that might be employed to control mosquito populations.

Nutritional and Fungal Load Dynamics of Fresh Brewers’ Grain Stored Under Aerobic Conditions

Brewers’ spent grain (BSG) is the amplest by-product of the brewing process. Fresh BSG is currently used as low-cost cattle feed due to its microbiological instability and high perishability. While recent research has looked the effects of storage time and temperature on the characteristics of wet brewers grains (WBG) as ruminant feeds. Three storage temperatures (15°C, 20°C, and 25°C) and periods (2, 4 and 6 days) were arranged in a 3×3 factorial design. Surface spoilage was not apparent at 15 °C throughout the storage periods. Deterioration was not also observed at 20 °C until the fourth day of storage where slight mold growth was apparent. Extensive mold growth was detected late in the sixth day at 20° C and continued manifestations up until the last day of storage at 25°C. Changes in major nutrients, DM losses, and yeast and mold colony count were significantly affected by the interaction of storage temperatures and durations (P<0.05). Except for samples stored at 15° C, nutrients contents decreased concomitantly (exceptions are ADF, lignin, and loss in DM) with prolonged storage times (p<0.05) and increasing temperatures (p<0.05). Contrast analysis indicated that it would be safe to store under aerobic storage conditions and feed the WBG for dairy cattle.

Total Aseptization of Boar Semen, to Increase the Biosecurity of Reproduction in Swine

The aim of the study was to establish the complete microbiological profile of boar semen (Sus scrofa domesticus) and to choose the most effective antiseptic measures in order to control and optimize AI reproduction in pig farms. One hundred and one semen samples were collected and analyzed from several pig farms. The microbiological profile of ejaculates was determined by evaluating the degree of contamination of fresh semen and after dilution with specific extenders. The bacterial and fungal load of fresh boar semen recorded an average value of 82.41/0.149 × 103 CFU/mL, while after diluting the ejaculates the contamination value was 0.354/0.140 × 103 CFU/mL. Twenty-four germs (15 bacterial and 9 fungal species) were isolated, the most common being Candida parapsilosis/sake (92%) and Escherichia coli (81.2%). Modification of the sperm collection protocol (HPBC) reduced contamination in raw sperm by 49.85% in bacteria (significant (p < 0.00001) and by 9.67% in fungi (non-significant (p < 0.111491). The load in bacteria and filamentous fungi can be controllable, but not in levuras fungi. Some fluconazole-added extenders (12.5 mg%), ensure fungal aseptization, and even an increase in sperm progressivity (8.39%) for at least a 12 h shelf life after dilution. Validation of sperm aseptization was done by maintaining sow fecundity unchanged after AI (insignificant p > 0.05).

Effectiveness Between Daily and After-Each-Case Room Disinfection of the Endoscopy Unit

Background: To evaluate the effectiveness between daily and after-each-case room disinfection in the endoscopy unit.Methods: This study was conducted in an endoscopy unit of the First Affiliation of Zhejiang Chinese Medical University. We cultured samples from the surface of endoscopy unit items, including operation unit air, isolation gown of an endoscopist, control panel buttons, workstation mouse, and the bed head of the patient. All the samples were divided into daily and after-each-case room disinfection groups. In addition, each group was subdivided into sedation and nonsedation gastroscopy with and without ventilation room groups.Results: The qualified rate of bed head samples of the patient were lower in the daily room disinfection group (76.67%) compared with the after-each-case group (100%). The isolation gown, mouse at the workstation, and the bed head of the patient demonstrated the lowest bacterial and fungal load in the after-each-case room disinfection group compared with the daily room disinfection group (p &lt; 0.05). In the subgroup analysis, a higher microbial load was observed for the isolation gown of the endoscopist used during nonsedation gastroscopy in an unventilated room under the after-each-case room disinfection pattern (p &lt; 0.05); a higher microbial load was observed for the control panel buttons used during nonsedation gastroscopy under the after-each-case room disinfection pattern (p &lt; 0.05).Conclusions: For risk-free or low-risk patients, daily room disinfection provides the basic health requirements of the endoscopy procedure. However, it is better to adopt the after-each-case room disinfection for the isolation gown of the endoscopist and bed head of the patient. For the patients with high risk, the after-each-case room disinfection is more suitable for every endoscopy unit (www.ClinicalTrials.gov, NCT04399005).

Association of LRRK2 rs11564258 single nucleotide polymorphisms with type and extent of gastrointestinal mycobiome in ulcerative colitis: a case–control study

Background Recently, the role of endogenous microbiota and the genotype-microbiota correlation in inflammatory bowel disease (IBD) pathogenesis have been highlighted. However, fungi, as the second most prevalent residents of the intestine, and their primary receptor, Dectin-1, are underrated. Thus, we conducted the first human study investigating the association of Leucine-rich repeat kinase 2 (LRRK2) polymorphism (rs11564258) with type and the extent of intestinal fungi in IBD patients. Material and methods A case–control study was performed on 79 ulcerative colitis (UC)-patients (case group) and 58 healthy subjects (HS group). DNA was extracted from blood samples of both groups and amplified with the primers designed for the specific locus containing the LRRK2 polymorphism (rs11564258) and then sequenced. Dectin-1 and LRRK2 mRNA expression levels were also determined. Furthermore, the type and prevalence of fecal yeast species were surveyed in case and control groups. Results A positive correlation was observed between rs11564258 polymorphism and UC susceptibility (p = 0.008 vs. HS). Patients with active UC had the highest rate of isolated fungal colonies (50.41%), followed by patients with non-active UC (24.6%) and HS (25%). These results showed a relationship between UC severity with the increased fungal load. Candida albicans had the highest prevalence in both UC (78.7%) and HS groups (55.8%). Whereas Saccharomyces cerevisiae was the second most common species detected in HS (15.23%), it was significantly reduced in the UC patient group (1.68%) (P = 0.0001). On the other hand, single nucleotide polymorphism (SNP, rs11564258) was not correlated with the increased fungal flora in the UC patients. The expression of LRRK2 and Dectin-1 mRNA detected in blood samples was notably higher in the UC patients (P < 0.01) than in the HS group, without being affected by rs11564258 polymorphism. Conclusions Here, we disclosed that LRRK2 mediates Dectin-1 signaling pathway activation and subsequent inflammation in the UC patients without being affected by the presence of SNP rs11564258. Our data showed an increased global fungal load in the UC patients along with elevated UC susceptibility in cases carrying rs11564258 polymorphism. However, more clinical investigations, particularly in larger populations with different ethnic groups, are required to support this conclusion.