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Keywords = formazan decomposition

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19 pages, 1390 KB  
Article
Heterotrophic Soil Microbes at Work: Short-Term Responses to Differentiated Fertilization Inputs
by Florin Aonofriesei, Alina Giorgiana Brotea (Andriescu) and Enuță Simion
Biology 2026, 15(1), 41; https://doi.org/10.3390/biology15010041 - 26 Dec 2025
Cited by 1 | Viewed by 885
Abstract
The interaction between organic and inorganic nutrients, bacterial communities, and soil fertility has been well documented over time. Conventional agricultural systems heavily utilize both inorganic and organic fertilizers, each exerting distinct effects on soil microbial dynamics and plant growth. The objective of our [...] Read more.
The interaction between organic and inorganic nutrients, bacterial communities, and soil fertility has been well documented over time. Conventional agricultural systems heavily utilize both inorganic and organic fertilizers, each exerting distinct effects on soil microbial dynamics and plant growth. The objective of our experiments was to identify the most effective fertilization strategy for improving the biological quality of a microbiologically impoverished and low-productivity soil. To this end, four fertilization strategies were evaluated: (i) organic fertilizers characterized by a high content of organic carbon (Fertil 4-5-7—variant 1); (ii) organic fertilizers with 12% organic nitrogen from proteins (Bio Ostara N—variant 2) (iii) combined inorganic–organic fertilizers (P35 Bio—variant 3) and (iv) mineral (inorganic) fertilizers (BioAktiv—variant V4). This study aimed to assess the short-term effects of fertilizers with varying chemical compositions on the density of cultivable heterotrophic bacteria and their associated dehydrogenase (DH) activity in a petrocalcic chernozem soil containing pedogenic carbonates. Soil sampling was conducted according to a randomized block design, comprising four replicates per treatment (control plus four fertilizer types). The enumeration of cultivable bacteria was performed using Nutrient Agar and A2R Agar media, whereas dehydrogenase activity (DHA) was quantified based on the reduction of 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) to 1,3,5-triphenyl-tetrazolium formazan (TPF) by bacterial dehydrogenase enzymes. Marked differences were observed in both parameters between the plots amended with inorganic fertilizers and those treated with organic fertilizers, as well as among the organic fertilizer treatments of varying composition. The most pronounced increases in both bacterial density and dehydrogenase activity (DHA) were recorded in the plots receiving the fertilizer with a high organic nitrogen content. In this treatment, the maximum bacterial population density reached 6.25 log10 CFU g−1 dry soil after approximately two months (May), followed by a significant decline starting in July. In contrast, DHA exhibited a more rapid response, reaching its peak in April (42.75 µg TPF g−1 soil), indicating an earlier DHA activation of microbial metabolism. This temporal lag between the two parameters suggests that enzymatic activity responded more swiftly to the nutrient inputs than did microbial biomass proliferation. For the other two organic fertilizer variants, bacterial population dynamics were broadly similar, with peak densities recorded in June, ranging from 5.98 log10 CFU g−1 soil (V3) to 6.03 log10 CFU g−1 soil (V1). A comparable trend was observed in DHA: in V3, maximum DHA was attained in June (30 µg TPF g−1 soil), after which it remained relatively stable, whereas in V1, it peaked in June (24.05 µg TPF g−1 soil) and subsequently declined slightly toward the end of the experimental period. Overall, the temporal dynamics of bacterial density and DHA demonstrated a strong dependence on the quality and biodegradability of the organic matter supplied by each fertilizer. Both parameters were consistently lower under inorganic fertilization compared with organic treatments, suggesting that the observed increases in microbial density and activity were primarily mediated by the enhanced availability of organic substrates. The relationship between the density of culturable heterotrophic bacteria and dehydrogenase (DH) activity was strongly positive (r = 0.79), indicating a close functional linkage between bacterial density and oxidative enzyme activity. This connection suggests that the culturable fraction of the heterotrophic microbial community plays a key role in the early stages of organic matter mineralization derived from the applied fertilizers, particularly in the decomposition of easily degradable substrates. Full article
(This article belongs to the Special Issue The Application of Microorganisms and Plants in Soil Improvement)
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15 pages, 2802 KB  
Article
Interfering with Color Response by Porphyrin-Related Compounds in the MTT Tetrazolium-Based Colorimetric Assay
by Bo Hee Choi, Mi-Ri Kim, Yu Na Jung, Smee Kang and Jungil Hong
Int. J. Mol. Sci. 2023, 24(1), 562; https://doi.org/10.3390/ijms24010562 - 29 Dec 2022
Cited by 12 | Viewed by 5943
Abstract
Porphyrin compounds are widely distributed in various natural products and biological systems. In this study, effects of porphyrin-related compounds including zinc protoporphyrin (ZnPP), protoporphyrin IX (PPIX), cyanocobalamin (CBL), hemin, and zinc phthalocyanine (ZnPC) were analyzed on color response of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) tetrazolium-based [...] Read more.
Porphyrin compounds are widely distributed in various natural products and biological systems. In this study, effects of porphyrin-related compounds including zinc protoporphyrin (ZnPP), protoporphyrin IX (PPIX), cyanocobalamin (CBL), hemin, and zinc phthalocyanine (ZnPC) were analyzed on color response of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) tetrazolium-based assay, a commonly-used method for analyzing cell viability. Color responses of MTT formazan formed in cells treated with ZnPP, PPIX, or ZnPC were significantly reduced even at submicromolar concentrations without affecting cell viability, whereas hemin and CBL did not. ZnPP, PPIX, and ZnPC rapidly induced degradation of MTT formazan already-produced by cells when exposed to light, but not under a dark condition. Photosensitizing properties of the three compounds were also verified through extensive generation of reactive oxygen species under light. The porphyrins did not affect the stability of water-soluble formazans including XTT, WST-1, WST-8, and MTS formazans. Several factors including different light sources and antioxidants modulated the degradation process of MTT formazan by the porphyrins. The results suggest that certain porphyrin compounds could cause a severe artifact in the MTT assay through rapid degradation of formazan dye due to their photosensitizing property, which needs to be considered carefully in the related assays. Full article
(This article belongs to the Special Issue Bioactive Compounds: Potential New Anti-inflammatory Drugs)
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19 pages, 669 KB  
Article
Case Study upon Foliar Application of Biofertilizers Affecting Microbial Biomass and Enzyme Activity in Soil and Yield Related Properties of Maize and Wheat Grains
by Dragana Latkovic, Jelena Maksimovic, Zoran Dinic, Radmila Pivic, Aleksandar Stanojkovic and Aleksandra Stanojkovic-Sebic
Biology 2020, 9(12), 452; https://doi.org/10.3390/biology9120452 - 8 Dec 2020
Cited by 39 | Viewed by 6268
Abstract
This study evaluated the effects of the application of microbial inoculants (N-fixing Klebsiella planticola and Enterobacter spp.), two rates of composite mineral fertilizers, and their combination on microbial biomass carbon (MBC), dehydrogenase (DHA), and proteinase activity (PTA) in Lessivated Cambisol and yield-related properties [...] Read more.
This study evaluated the effects of the application of microbial inoculants (N-fixing Klebsiella planticola and Enterobacter spp.), two rates of composite mineral fertilizers, and their combination on microbial biomass carbon (MBC), dehydrogenase (DHA), and proteinase activity (PTA) in Lessivated Cambisol and yield-related properties of maize and wheat grains in a two-year trial. Unfertilized soil was used as a control variant. MBC was measured using the chloroform fumigation-extraction method, DHA was determined spectrophotometrically by measuring the intensity of the formed red-colored triphenyl formazan, while PTA was determined using a titration method by measuring the degree of gelatine decomposition. In grain samples, P was determined spectrophotometrically, K—by flame emission photometry, N—on an elemental carbon/nitrogen/sulfur (CNS) analyzer, and crude proteins—by calculation of N content. Measuring both crops’ yield was carried out at the end of the vegetation. The results indicated that mineral fertilizers are not, in general, negative for soil microbiota when used in the context of sustainable agriculture without monoculture. There is a significant increase in the values of soil MBC, DHA, and PTA in the variants with combined application of bacterial inoculants and lower rates of mineral fertilizers. The highest values of these parameters were determined in the period with a better distribution of precipitation during the vegetation period of the year. The mentioned combination also resulted in a higher grain yield of maize and wheat comparing to the application of lower rates of the NPK nutrients solely. The combined application of high rates of mineral fertilizers and bacterial inoculants resulted in significantly increased N, P, K, and protein content in the grains of crops, and the same applied to yield. Concluding, studied bacterial inoculants can be used to specify the replacement of nitrogen fertilizers, stimulating the microbial biomass and enzyme activity in the soil, helping to ensure that the supply of nutrients contributing to an optimized yield of crops is maintained. Full article
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