Comprehensive assessment of goat adipose tissue-derived mesenchymal stem cells cultured in different media.

Mesenchymal stem cells (MSCs) have demonstrated potential in treating livestock diseases that are unresponsive to conventional therapies. MSCs derived from goats, a valuable model for studying orthopaedic disorders in humans, offer insights into bone formation and regeneration. Adipose tissue-derived MSCs (ADSCs) are easily accessible and have a high capacity for expansion. Although the choice of culture media significantly influences the biological properties of MSCs, the optimal media for goat ADSCs (gADSCs) remains unclear. This study aimed to assess the effects of four commonly used culture media on gADSCs' culture characteristics, stem cell-specific immunophenotype, and differentiation. Results showed that MEM, DMEM/F12, and DMEM-LG were superior in maintaining cell morphology and culture parameters of gADSCs, such as cell adherence, metabolic activity, colony-forming potential, and population doubling. Conversely, DMEM-HG exhibited poor performance across all evaluated parameters. The gADSCs cultured in DMEM/F12 showed enhanced early proliferation and lower apoptosis. The cell surface marker distribution exhibited superior characteristics in gADSCs cultured in MEM and DMEM/F12. In contrast, the distribution was inferior in gADSCs cultured in DMEM-LG. DMEM/F12 and DMEM-LG culture media demonstrated a significantly higher potential for chondrogenic differentiation and DMEM-LG for osteogenic differentiation. In conclusion, DMEM/F12 is a suitable culture medium for propagating gADSCs as it effectively maintains cell morphology, growth parameters, proliferation and lower apoptosis while exhibiting desirable expression patterns of MSC-specific markers. These findings contribute to optimising culture conditions for gADSCs, enhancing their potential applications in disease treatment and regenerative medicine.

Pilot-scale production of Bacillus subtilis MSCL 897 spore biomass and antifungal secondary metabolites in a low-cost medium.

OBJECTIVES: Bacillus subtilis is a plant growth promoting bacterium (PGPB) that acts as a microbial fertilizer and biocontrol agent, providing benefits such as boosting crop productivity and improving nutrient content. It is able to produce secondary metabolites and endospores simultaneously, enhancing its ability to survive in unfavorable conditions and eliminate competing microorganisms. Optimizing cultivation methods to produce B. subtilis MSCL 897 spores on an industrial scale, requires a suitable medium, typically made from food industry by-products, and optimal temperature and pH levels to achieve high vegetative cell and spore densities with maximum productivity. RESULTS: This research demonstrates successful pilot-scale (100 L bioreactor) production of a biocontrol agent B. subtilis with good spore yields (1.5 × 109 spores mL-1) and a high degree of sporulation (>80%) using a low-cost cultivation medium. Culture samples showed excellent antifungal activity (1.6-2.3 cm) against several phytopathogenic fungi. An improved methodology for inoculum preparation was investigated to ensure an optimal seed culture state prior to inoculation, promoting process batch-to-batch repeatability. Increasing the molasses concentration in the medium and operating the process in fed-batch mode with additional molasses feed, did not improve the overall spore yield, hence, process operation in batch mode with 10 g molasses L-1 is preferred. Results also showed that the product quality was not significantly impacted for up to 12 months of storage at room temperature. CONCLUSION: An economically-feasible process for B. subtilis-based biocontrol agent production was successfully developed at the pilot scale.

Life cycle assessment and technoeconomic analysis of biofuels produced from polyculture microalgae cultivated in greywater.

This study evaluated the environmental and economic impacts of substituting synthetic media with greywater for cultivating microalgae in the biofuel production process. Life cycle assessment (LCA) and technoeconomic assessment (TEA) were employed to compare the impacts of two scenarios - one containing bold's basal (BB) media and another containing greywater as growth mediums for microalgae cultivation. Scenarios 1 and 2 mitigated 1.74 and 2.14 kg CO2 per kg of biofuel production, respectively. Substituting BB media with greywater resulted in a 16.3% reduction in energy requirements, leading to a 79.3% increase in net energy recovered. LCA findings demonstrate a reduction in all seven environmental categories. TEA reveals that, despite a 21.7% higher capital investment, scenario 2 proves more economically viable due to a 39.8% lower operating cost and additional revenue from wastewater treatment and carbon credits. The minimum selling price of biofuel dropped from Rs 73.5/kg to Rs 36.5/kg, highlighting the economic and environmental advantages of substituting BB media with greywater in microalgal biofuel production.

Assessment of UV radiation effects on airborne mucormycetes and bacterial populations in a hospital environment.

Infections, such as mucormycosis, often result from inhaling sporangiospore present in the environment. Surprisingly, the extent of airborne Mucormycetes sporangiospore concentrations remains inadequately explored. This study aimed to assess the influence of UV radiation on microbial populations and Mucormycetes spore levels within a hospital environment in northern Iran. A comprehensive dataset comprising 298 air samples collected from both indoor and outdoor settings was compiled. The culture was conducted using Blood Agar and Dichloran Rose Bengal Chloramphenicol (DRBC) culture media, with Chloramphenicol included for fungal agents and Blood Agar for bacterial. Before UV treatment, the average count of Mucormycetes ranged from 0 to 26.4 ± 25.28 CFU m-3, fungal agents from 2.24 ± 3.22 to 117.24 ± 27.6 CFU m-3, and bacterial agents from 29.03 ± 9.9 to 359.37 ± 68.50 CFU m-3. Following UV irradiation, the averages were as follows: Mucormycetes ranged from 0 to 7.85 ± 6.8 CFU m-3, fungal agents from 16.58 ± 4.79 to 154.98 ± 28.35 CFU m-3, and bacterial agents from 0.38 ± 0.65 to 43.92 ± 6.50 CFU m-3. This study, notably marks the pioneering use of UV light to mitigate Mucormycetes spore counts and bacterial agents in northeastern Iran, contributing to the advancement of environmental health and safety practices in hospital settings.

Growth assessment of Salmonella enterica multi-serovar populations in poultry rinsates with commonly used enrichment and plating media.

Isolation of Salmonella from enrichment cultures of food or environmental samples is a complicated process. Numerous factors including fitness in various selective enrichment media, relative starting concentrations in pre-enrichment, and competition among multi-serovar populations and associated natural microflora, come together to determine which serovars are identified from a given sample. A recently developed approach for assessing the relative abundance (RA) of multi-serovar Salmonella populations (CRISPR-SeroSeq or Deep Serotyping, DST) is providing new insight into how these factors impact the serovars observed, especially when different selective enrichment methods are used to identify Salmonella from a primary enrichment sample. To illustrate this, we examined Salmonella-positive poultry pre-enrichment samples through the selective enrichment process in Tetrathionate (TT) and Rappaport Vassiliadis (RVS) broths and assessed recovery of serovars with each medium. We observed the RA of serovars detected post selective enrichment varied depending on the medium used, initial concentration, and competitive fitness factors, all which could result in minority serovars in pre-enrichment becoming dominant serovars post selective enrichment. The data presented provide a greater understanding of culture biases and lays the groundwork for investigations into robust enrichment and plating media combinations for detecting Salmonella serovars of greater concern for human health.

Global patterns of human exposure to flame retardants indoors.

To fill the knowledge gaps regarding the global patterns of human exposure to flame retardants (FRs) (i.e., brominated flame retardants (BFRs) and organophosphorus flame retardants (OPFRs)), data on the levels and distributions of FRs in external and internal exposure mediums, including indoor dust, indoor air, skin wipe, serum and urine, were summarized and analysed. Comparatively, FR levels were relatively higher in developed regions in all mediums, and significant positive correlations between FR contamination and economic development level were observed in indoor dust and air. Over time, the concentration of BFRs showed a slightly decreasing trend in all mediums worldwide, whereas OPFRs represented an upward tendency in some regions (e.g., the USA and China). The occurrence levels of FRs and their metabolites in all external and internal media were generally correlated, implying a mutual indicative role among them. Dermal absorption generally contributed >60% of the total exposure of most FR monomers, and dust ingestion was dominant for several low volatile compounds, while inhalation was found to be negligible. The high-risk FR monomers (BDE-47, BDE-99 and TCIPP) identified by external exposure assessment showed similarity to the major FRs or metabolites observed in internal exposure mediums, suggesting the feasibility of using these methods to characterize human exposure and the contribution of indoor exposure to the human burden of FRs. This review highlights the significant importance of exposure assessment based on multiple mediums for future studies.

Assessment of the impact of centralized bioMérieux BACT/ALERT® VIRTUO® blood culture system (VIRTUO) implementation on outcomes in patients with gram-negative bacteremia.

BACKGROUND: We evaluated pre- and postimplementation of Virtuo on outcome in patients with gram-negative bacteremia using a quasiexperimental time-in-motion design. METHODS: Becton Dickinson BACTEC™ 9000 series (Bactec) (2018) and Virtuo system (2020) were utilized in a decentralized and centralized process, respectively. Data collected in August-December in 2018 and 2020 were analyzed with SPSS (ver 28). RESULTS: For 185 patients in each time period, patient age, gender, length of hospitalization were not different. However, blood culture (BC) volume was significantly lower in 2020 (7.1 ± 2.6 mL) compared to 2018 (8.9 ± 1.9 mL). Time from BC draw and time from pathogen identification (ID) to treatment change were both significantly faster in 2020 (52.9 ± 38.3 hours; 15.1 ± 27.4 hours), compared to 2018 (65.0 ± 46.3 hours; 23.8 ± 33.8), respectively. CONCLUSIONS: Replacement of decentralized Bactec with centralized Virtuo, resulted in significant improvement in management of patients gram-negative bacteremia.

Trends and patterns of polycyclic aromatic hydrocarbons (PAHs) in forest fire-affected soils and water mediums with implications on human health risk assessment.

Forest fires are extreme natural/artificial events releasing polycyclic aromatic hydrocarbons (PAHs), which are carcinogenic. Most of the released PAHs are trapped in burnt ash, a part of which is transported and settle on different mediums like soil and water. After strong rainfall events, PAHs enter into surface water bodies through surface runoff, thereby deteriorating water quality. Changes in PAHs levels during the post-fire duration and human health risks due to PAHs released from forest fires need attention. This study aim to explain the trends and patterns of PAHs and health risks due to exposure to soil and water contaminated with PAHs. Forest fires release a higher percentage of low molecular weight PAHs (LMW PAHs) than high molecular weight PAHs (HMW PAHs). Ash and burnt soils contain a higher percentage of LMW PAHs since biomass burning releases huge amounts of LMW PAHs. Whereas, sediments contain a higher percentage of HMW PAHs since most of the LMW PAHs are already degraded. HMW PAHs were causing higher risk to humans (both cancer and non-cancer) due to their higher oxidation potential. Exposure to water contaminated by PAHs resulted in higher health risks for both BaP equivalent and a mixture of PAHs. Exposure to sediment produced the highest health risk due to a higher percentage of HMW PAHs, followed by surface water, burnt soil, ash, and unburnt soil. Cancer and non-cancer risk due to dermal exposure was more elevated than oral exposure. The mixture of PAHs in sediment produced a higher average dermal risk for children (2.21E+00 for cancer and 7.69E+03 for non-cancer risk) and oral cancer risk for adults (7.11E-03). However, exposure to BaP equivalent in sediment produced higher oral non-cancer risk (7.01E+02) for children. Thus, effective PAHs monitoring is required in both soil and surface water mediums for ensuring proper treatment in water supply systems.

Cell culture media dependent in vitro dynamics and culture characteristics of adult caprine dermal fibroblast cells.

The enhanced availability of functional fibroblasts from precious tissue samples requires an ideal cell-culture system. Therefore, this study was designed to investigate the performance of caprine adult fibroblast cells (cadFibroblast) when cultivated in different culture media. The cadFibroblast cell lines from adult Barbari (Capra hircus) bucks were established and the effect of different media viz. DMEM/F-12 [with low-glucose (5.5 mM; DL) and high-glucose (30 mM; DH)], α-MEM [with low-glucose (5.5 mM; ML) and with high-glucose (30 mM; MH)], and fibroblast growth medium (FGM) were evaluated. Cells were then compared for growth characteristics and in-vitro dynamics through cellular morphology, proliferation, population-doubling time, double-immunocytochemistry, colony-forming units, wound healing, transwell migration, and differential expression of fibroblast-specific markers (FSP-1 and vimentin). The results of immunocytochemistry, transwell migration/invasion, and wound healing assays showed the superiority of DH over DL and other media tested. Whereas, similar effects of glucose supplementation and expression of FSP-1 were not observed in α-MEM. Transwell migration was significantly (p < 0.05) lower in FGM compared with other media tested. Overall, our results illustrate the media-dependent deviation in in-vitro dynamics and culture characteristics of cadFibroblasts that may be useful to develop strategies to cultivate these cells efficiently for research and downstream applications.

Smart Detecting and Versatile Wearable Electrical Sensing Mediums for Healthcare.

A rapidly expanding global population and a sizeable portion of it that is aging are the main causes of the significant increase in healthcare costs. Healthcare in terms of monitoring systems is undergoing radical changes, making it possible to gauge or monitor the health conditions of people constantly, while also removing some minor possibilities of going to the hospital. The development of automated devices that are either attached to organs or the skin, continually monitoring human activity, has been made feasible by advancements in sensor technologies, embedded systems, wireless communication technologies, nanotechnologies, and miniaturization being ultra-thin, lightweight, highly flexible, and stretchable. Wearable sensors track physiological signs together with other symptoms such as respiration, pulse, and gait pattern, etc., to spot unusual or unexpected events. Help may therefore be provided when it is required. In this study, wearable sensor-based activity-monitoring systems for people are reviewed, along with the problems that need to be overcome. In this review, we have shown smart detecting and versatile wearable electrical sensing mediums in healthcare. We have compiled piezoelectric-, electrostatic-, and thermoelectric-based wearable sensors and their working mechanisms, along with their principles, while keeping in view the different medical and healthcare conditions and a discussion on the application of these biosensors in human health. A comparison is also made between the three types of wearable energy-harvesting sensors: piezoelectric-, electrostatic-, and thermoelectric-based on their output performance. Finally, we provide a future outlook on the current challenges and opportunities.

Low-cost culture media designed for biomass production of beneficial lactic acid bacteria for their inclusion in a formula to treat bovine reproductive infections.

As a first step for the use of probiotics in a formula for cattle, it is required to have available low-cost culture medium(s) and efficient production conditions for the growth of probiotic bacteria and high production of cell biomass. De Man-Rogosa-Sharpe medium, used frequently for lactic acid bacteria (LAB) contains adequate ingredients for their growth but is very expensive for industrial application. The nutrients required for LAB growth are strain-dependent. In this work, traditional culture media were evaluated omitting and/or modifying ingredients in their composition, as carbon or nitrogen source, on the basis of their low-cost industrial waste, to select those supporting the most efficient growth. The results showed that the formulation of culture media containing fructose (0.5%) and molasses (1.0%) was better for the growth and production of cell biomass for all the strains assayed, except Lactobacillus gasseri CRL1421 growing in 1.5% corn syrup. FM902 yeast extract at concentrations between 1.5% and 2.5% was the most adequate for most of the strains. The LAB grown in the designed media maintained the beneficial properties for which they selected. The use of the culture media designed to produce biomass decrease production costs, which is an important step for the feasible industrial production of probiotic pharmaceuticals.

Assessment of Omitting MacConkey Agar as a Primary Inoculating Medium for MALDI-TOF MS-Based Bacterial Identification from Urine, Blood, and Respiratory Samples.

OBJECTIVE: MacConkey agar (MAC) is commonly used as a primary medium for conventional bacterial identification in clinical microbiology laboratories. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has revolutionized microbial identification and is considered a reliable identification tool. While conventional identification methods rely on colony characteristics, MALDI-TOF MS requires a pure isolate on a solid medium. METHODS: This study investigated whether MAC can be omitted as a routine inoculation medium for urine, lower respiratory tract (LRT), and positive blood culture samples. The study included 462 clinical samples. Among these, 221 were urine samples, 141 were positive blood cultures, and the remaining 100 were LRT samples. The samples were inoculated on blood agar (BA) and MAC for the control group and on BA only for the experimental group, followed by incubation and identification with MALDI-TOF MS. RESULTS: The BA only group showed the same microbial identification using MALDI-TOF MS as the control BA and MAC groups for blood and LRT specimens. For urine samples, 99.1% (219/221) of the samples produced the same identification results for the two groups. The cause of discrepant results for two urine specimens was due to Proteus species overgrowth on BA, which hindered non-Proteus spp. identification for the BA-only group. CONCLUSION: Our results may indicate that omitting MAC has little or no impact on the recovery of organisms present in culture. However, due to possible Proteus spp. overgrowth, caution should be exercised in the decision to omit MAC from the primary inoculating medium, which necessitates further studies in other centers with a larger sample size.

Reliable assessment of carbon black nanomaterial of a variety of cell culture media for in vitro toxicity assays by asymmetrical flow field-flow fractionation.

Carbon black nanomaterial (CB-NM), as an industrial product with a large number of applications, poses a high risk of exposure, and its impact on health needs to be assessed. The most common testing platform for engineered (E)NMs is in vitro toxicity assessment, which requires prior ENM dispersion, stabilization, and characterization in cell culture media. Here, asymmetric flow field-flow fractionation (AF4) coupled to UV-Vis and dynamic light scattering (DLS) detectors in series was used for the study of CB dispersions in cell culture media, optimizing instrumental variables and working conditions. It was possible to disperse CB in a non-ionic surfactant aqueous solution due to the steric effect provided by surfactant molecules attached on the CB surface which prevented agglomeration. The protection provided by the surfactant or by culture media alone was insufficient to ensure good dispersion stability needed for carrying out in vitro toxicity studies. On the other hand, cell culture media in combination with the surfactant improved dispersion stability considerably, enabling the generation of shorter particles and a more favourable zeta potential magnitude, leading to greater stability due to electrostatic repulsion. It was demonstrated that the presence of amino acids in the culture media improved the monodisperse nature and stability of the CB dispersions, and resulted in a turn towards more negative zeta potential values when the pH was above the amino acid isoelectric point (IEP). Culture media used in real cell culture scenarios were also tested, and in vitro toxicity assays were developed optimizing the compatible amount of surfactant.

Cost-Efficient Production of the Sphingan WL Gum by Sphingomonas sp. WG Using Molasses and Sucrose as the Carbon Sources.

The polysaccharide WL gum is produced by the marine microorganism Sphingomonas sp. WG and presents great commercial utility potential in many industries especially in oil industries. However, the high fermentation cost limits its wide application. Therefore, an efficient production system at a lower cost was established using beet molasses to partially replace the commonly used carbon sources. Four different molasses were screened and their composition was investigated. One-factor design and RSM statistical analysis were employed to optimize the WL gum fermentation medium. The effects of molasses on the rheological properties and gene expression of WL gum were also investigated. The results showed that the pretreated beet molasses generated both high broth viscosity and WL gum production (12.94 Pa·s and 11.16 g/L). Heavy metal ions and ash were found to be the key factors in unpretreated and pretreated molasses affecting WL production. The cost-efficient production medium contained (g/L): sucrose 61.79, molasses 9.95, yeast extract 1.23, K2HPO4 1, MgSO4 0.1, ZnSO4 0.1 and the WL gum production reached 40.25 ± 1.15 g/L. The WL gum product WL-molasses showed the higher apparent viscosity, and viscous modulus and elastic modulus than WL-sucrose and WL-mix, which might be related to its highest molecular mass. The higher expressional level of genes such as pgm, ugp, ugd, rmlA, welS, and welG in WL gum synthesis in the mixed carbon source medium caused the high production and broth viscosity. This work provided a cost-efficient method for WL gum production.

Economical Optimization of Industrial Medium Culture for Bacterial Cellulose Production.

The competitiveness of bacterial cellulose (BC) production with plant cellulose can be achieved by production on cost-effective media. It was found that the bacterial cell number ratio of BC to culture medium increases over time so that from the fourth day, the entrapped cell number in the cellulose network exceeds the suspended cells. Optimization based on 23-full factorial showed that inoculum development at 50 rpm and the main culture process under static conditions significantly increases BC production. A cost-effective culture medium containing molasses (ML) and corn steep liquor (CSL) was developed based on the same C/N ratio to HS medium, with 7.24 g/l cellulose at C/N ratio 12.6 is competitive with maximum production 8.7 g/L in HS medium. The BC production cost was reduced about 94% using the proposed cheap and locally available medium containing ML and CSL, while BC mechanical properties increased by about 50%.

Evaluation of three alternatives cost-effective culture media for Mycobacterium tuberculosis detection and drug susceptibility determination using the microscopic observation drug susceptibility (MODS) assay.

Tuberculosis phenotypic detection assays are commonly used in low-resource countries. Therefore, reliable detection methods are crucial for early diagnosis and treatment. The microscopic observation drug susceptibility (MODS) assay is a culture-based test to detect Mycobacterium tuberculosis and characterize drug resistance in 7-10 days directly from sputum. The use of MODS is limited by the availability of supplies necessary for preparing the enriched culture. In this study, we evaluated three dry culture media that are easier to produce and cheaper than the standard one used in MODS [1]: an unsterilized powder-based mixed (Boldú et al., 2007) [2], a sterile-lyophilized medium, and (Sengstake et al., 2017) [3] an irradiated powder-based mixed. Mycobacterial growth and drug susceptibility were evaluated for rifampin, isoniazid, and pyrazinamide (PZA). The alternative cultures were evaluated using 282 sputum samples with positive acid-fast smears. No significant differences were observed in the positivity test rates. The positivity time showed high correlations (Rho) of 0.925, 0.889, and 0.866 between each of the three alternative media and the standard. Susceptibility testing for MDR and PZA showed an excellent concordance of 1 compared to the reference test. These results demonstrate that dry culture media are appropriate and advantageous for use in MODS in low-resource settings.

Analysis of reproducibility and robustness of OrganoPlate® 2-lane 96, a liver microphysiological system for studies of pharmacokinetics and toxicological assessment of drugs.

Establishing the functionality, reproducibility, robustness, and reliability of microphysiological systems is a critical need for adoption of these technologies. A high throughput microphysiological system for liver studies was recently proposed in which induced pluripotent stem cell-derived hepatocytes (iHeps) and non-parenchymal cells (endothelial cells and THP-1 cells differentiated with phorbol 12-myristate 13-acetate into macrophage-like cells) were co-cultured in OrganoPlate® 2-lane 96 devices. The goal of this study was to evaluate this platform using additional cell types and conditions and characterize its utility and reproducibility. Primary human hepatocytes or iHeps, with and without non-parenchymal cells, were cultured for up to 17 days. Image-based cell viability, albumin and urea secretion into culture media, CYP3A4 activity and drug metabolism were assessed. The iHeps co-cultured with non-parenchymal cells demonstrated stable cell viability and function up to 17 days; however, variability was appreciable both within and among studies. The iHeps in monoculture did not form clusters and lost viability and function over time. The primary human hepatocytes in monoculture also exhibited low cell viability and hepatic function. Metabolism of various drugs was most efficient when iHeps were co-cultured with non-parenchymal cells. Overall, we found that the OrganoPlate® 2-lane 96 device, when used with iHeps and non-parenchymal cells, is a functional liver microphysiological model; however, the high-throughput nature of this model is somewhat dampened by the need for replicates to compensate for high variability.

Optimization of in vitro culture of honeybee nervous tissue for pesticide risk assessment.

The most used pesticides have neurotoxic action on the neurotransmitter system of target and non-targeted insects, such as honeybees. However, honeybees have foremost importance worldwide, which has encouraged the development of tools to evaluate the action of specific pesticide molecules on their nervous system, providing accurate data on damage to their brain. In this sense, our study aimed to optimize in vitro honeybee nervous tissue culture to assess pesticide risks. To this end, six forager honeybee brains were dissected and transferred to different combinations of Leibovitz-15 (L-15) culture medium supplemented with Fetal Bovine Serum (FBS), Hank's Balanced Salt Solution (HBSS), and Insect Medium Supplement (IMS). Nervous tissues were collected after different incubation times (1, 6, 12, and 24 h) for morphology and Kenyon cell analyses. Our results showed that L-15 medium supplemented with HBSS and with HBSS plus FBS were the best media for culturing honey nervous tissue for 24 h, as they resulted in less tissue spacing and cell disarrangement. Therefore, they may be assessed in future ecotoxicological tests.

Assessment of the Potential of a Native Non-Aflatoxigenic Aspergillus flavus Isolate to Reduce Aflatoxin Contamination in Dairy Feed.

Aspergillus species can produce aflatoxins (AFs), which can severely affect human and animal health. The objective was to evaluate the efficacy of reducing AF contamination of a non-aflatoxigenic isolate of A. flavus experimentally coinoculated with different aflatoxigenic strains in whole plant (WP), corn silage (CS), immature grains (IG) and in culture media (CM). An L-morphotype of A. flavus (CS1) was obtained from CS in a dairy farm located in the Mexican Highland Plateau; The CS1 failed to amplify the AFs biosynthetic pathway regulatory gene (aflR). Monosporic CS1 isolates were coinoculated in WP, CS, IG and CM, together with A. flavus strains with known aflatoxigenic capacity (originating from Cuautitlán and Tamaulipas, Mexico), and native isolates from concentrate feed (CF1, CF2 and CF3) and CS (CS2, CS3). AF production was evaluated by HPLC and fungal growth rate was measured on culture media. The positive control strains and those isolated from CF produced a large average amount of AFs (15,622 ± 3952 and 12,189 ± 3311 µg/kg), whereas A. flavus strains obtained from CS produced a lower AF concentration (126 ± 25.9 µg/kg). CS1 was efficient (p < 0.01) in decreasing AF concentrations when coinoculated together with CF, CS and aflatoxigenic positive control strains (71.6−88.7, 51.0−51.1 and 63.1−71.5%) on WP, CS, IG and CM substrates (73.9−78.2, 65.1−73.7, 63.8−68.4 and 57.4−67.6%). The results suggest that the non-aflatoxigenic isolate can be an effective tool to reduce AF contamination in feed and to minimize the presence of its metabolites in raw milk and dairy products intended for human nutrition.

Towards large scale biocrust restoration: Producing an efficient and low-cost inoculum of N-fixing cyanobacteria.

Dryland soil degradation is increasing due to global change and traditional restoration methods are not successful due to water scarcity. Thus, an alternative technology based on inoculating biocrust-forming cyanobacteria on degraded soils has emerged. Biocrusts are communities of mosses, lichens, cyanobacteria or fungi that colonize soil surface forming a stable and fertile layer. Previous studies have shown the benefits of inoculating cyanobacteria to restore soils at a small scale. However, to face field restoration projects, it is necessary to produce high quantities of biomass at an affordable cost. In this work, we analyze if the previously tested cyanobacteria Scytonema hyalinum, Tolypothrix distorta (heterocystous strains) and Trichocoleus desertorum (a bundle-forming one) can be produced with agricultural fertilizers. Different culture media were used: two containing pure chemicals (BG11 and BG110, this N-free medium was used just for heterocystous strains) and two containing fertilizers (BG11-F and MM-F). The performance of the cultures was monitored by measuring the biomass concentration and photosynthetic stress. Afterwards, we analyzed their capacity to induce biocrusts and improve soil properties by inoculating the biomass on a mine substrate indoors and measuring, three months later, the albedo, chlorophyll a and organic carbon content. Results show that the bundle-forming cyanobacterium was unable to grow in the media tested, whereas both heterocystous cyanobacteria grew in all of them and induced the formation of biocrusts improving the organic carbon substrate content. The best results for S. hyalinum were found using the MM-F medium, and for T. distorta using a medium containing pure chemicals (BG11). However, results were also positive when using a medium containing fertilizers (BG11-F). Thus, agricultural fertilizers can be used to undertake the production of heterocystous cyanobacteria for large scale restoration in drylands. On the other hand, more research is needed to find sustainable techniques to produce biomass of bundle-forming cyanobacteria.