Multifaced application of AFEX-pretreated biomass in producing second-generation biofuels, ruminant animal feed, and value-added bioproducts.

Lignocellulosic biomass holds a crucial position in the prospective bio-based economy, serving as a sustainable and renewable source for a variety of bio-based products. These products play a vital role in displacing fossil fuels and contributing to environmental well-being. However, the inherent recalcitrance of biomass poses a significant obstacle to the efficient access of sugar polymers. Consequently, the bioconversion of lignocellulosic biomass into fermentable sugars remains a prominent challenge in biorefinery processes to produce biofuels and biochemicals. In addressing these challenges, extensive efforts have been dedicated to mitigating biomass recalcitrance through diverse pretreatment methods. One noteworthy process is Ammonia Fiber Expansion (AFEX) pretreatment, characterized by its dry-to-dry nature and minimal water usage. The volatile ammonia, acting as a catalyst in the process, is recyclable. AFEX contributes to cleaning biomass ester linkages and facilitating the opening of cell wall structures, enhancing enzyme accessibility and leading to a fivefold increase in sugar conversion compared to untreated biomass. Over the last decade, AFEX has demonstrated substantial success in augmenting the efficiency of biomass conversion processes. This success has unlocked the potential for sustainable and economically viable biorefineries. This paper offers a comprehensive review of studies focusing on the utilization of AFEX-pretreated biomass in the production of second-generation biofuels, ruminant feed, and additional value-added bioproducts like enzymes, lipids, proteins, and mushrooms. It delves into the details of the AFEX pretreatment process at both laboratory and pilot scales, elucidates the mechanism of action, and underscores the role of AFEX in the biorefinery for developing biofuels and bioproducts, and nutritious ruminant animal feed production. While highlighting the strides made, the paper also addresses current challenges in the commercialization of AFEX pretreatment within biorefineries. Furthermore, it outlines critical considerations that must be addressed to overcome these challenges, ensuring the continued progress and widespread adoption of AFEX in advancing sustainable and economically viable bio-based industries.

Long-term dietary exposure to the non-steroidal anti-inflammatory drugs diclofenac and ibuprofen can affect the physiology of common carp (Cyprinus carpio) on multiple levels, even at "environmentally relevant" concentrations.

The aim of the study was to evaluate the effects of emerging environmental contaminants, the non-steroidal anti-inflammatory drugs (NSAIDs) diclofenac (DCF) and ibuprofen (IBP), on physiological functions in juvenile common carp (Cyprinus carpio). Fish were exposed for 6 weeks, and for the first time, NSAIDs were administered through diet. Either substance was tested at two concentrations, 20 or 2000 µg/kg, resulting in four different treatments (DCF 20, DCF 2000, IBP 20, IBP 2000). The effects on haematological and biochemical profiles, the biomarkers of oxidative stress, and endocrine disruption were studied, and changes in RNA transcription were also monitored to obtain a comprehensive picture of toxicity. Fish exposure to high concentrations of NSAIDs (DCF 2000, IBP 2000) elicited numerous statistically significant changes (p < 0.05) in the endpoints investigated, with DCF being almost always more efficient than IBP. Compared to control fish, a decrease in total leukocyte count attributed to relative lymphopenia was observed. Plasma concentrations of total proteins, ammonia, and thyroxine, and enzyme activities of alanine aminotransferase (ALT), aspartate aminotransferase, and alkaline phosphatase (ALP) were significantly elevated in either group, as were the activities of certain hepatic antioxidant enzymes (superoxide dismutase, glutathione-S-transferase) in the DCF 2000 group. The transcriptomic profile of selected genes in the tissues of exposed fish was affected as well. Significant changes in plasma total proteins, ammonia, ALT, and ALP, as well as in the transcription of genes related to thyroid function and the antioxidant defense of the organism, were found even in fish exposed to the lower DCF concentration (DCF 20). As it was chosen to match DCF concentrations commonly detected in aquatic invertebrates (i.e., the potential feed source of fish), it can be considered "environmentally relevant". Future research is necessary to shed more light on the dietary NSAID toxicity to fish.

Assessing the effectiveness of 3, 4-dimethylpyrazole phosphate (DMPP) inhibitor in mitigating N2O emissions from contrasting Cd-contaminated soils.

Improving nitrogen use efficiency of chemical fertilizers is essential to mitigate the negative environmental impacts of nitrogen. Nitrification, the conversion of ammonium to nitrate via nitrite by soil microbes, is a prominent source of nitrogen loss in soil systems. The effectiveness of nitrification inhibitors in reducing nitrogen loss through inhibition of nitrification is well-documented, however, their efficacy in heavy metals-contaminated soils needs thorough investigations. The current study assessed the efficacy of nitrification inhibitor 3, 4-dimethylpyrazole phosphate (DMPP) in reducing nitrous oxide (N2O) emissions in cadmium (Cd) contaminated paddy and red soils under lab-controlled environment. Obtained results indicated the substantial reduction in N2O emissions with DMPP in paddy and red soil by 48 and 35 %, respectively. However, Cd contamination resulted in reduced efficacy of DMPP, thus decreased the N2O emissions by 36 and 25 % in paddy and red soil, respectively. It was found that addition of DMPP had a significant effect on the abundance of ammonia oxidizing bacteria (AOB) and archaea (AOA). Notably, the reduction in N2O emissions by DMPP varied with the abundance of AOB. Moreover, Cd pollution resulted in a significant (P < 0.05) reduction in the abundance of archaeal and bacterial amoA genes, as well as bacterial nirK, nirS, and nosZ genes. The combined treatment of Cd and DMPP had a detrimental impact on denitrifiers, thereby influencing the overall efficiency of DMPP. These findings provide novel insights into the application of DMPP to mitigate nitrification and its potential role in reducing N2O emissions in contaminated soils.

H2 supplied via ammonia borane stimulates lateral root branching via phytomelatonin signaling.

A reliable and stable hydrogen gas (H2) supply will benefit agricultural laboratory and field trials. Here, we assessed ammonia borane (AB), an efficient hydrogen storage material used in the energy industry, and determined its effect on plant physiology and the corresponding mechanism. Through hydroponics and pot experiments, we discovered that AB increases tomato (Solanum lycopersicum) lateral root (LR) branching and this function depended on the increased endogenous H2 level caused by the sustainable H2 supply. In particular, AB might trigger LR primordia initiation. Transgenic tomato and Arabidopsis (Arabidopsis thaliana) expressing hydrogenase1 (CrHYD1) from Chlamydomonas reinhardtii not only accumulated higher endogenous H2 and phytomelatonin levels but also displayed pronounced LR branching. These endogenous H2 responses achieved by AB or genetic manipulation were sensitive to the pharmacological removal of phytomelatonin, indicating the downstream role of phytomelatonin in endogenous H2 control of LR formation. Consistently, extra H2 supply failed to influence the LR defective phenotypes in phytomelatonin synthetic mutants. Molecular evidence showed that the phytomelatonin-regulated auxin signaling network and cell-cycle regulation were associated with the AB/H2 control of LR branching. Also, AB and melatonin had little effect on LR branching in the presence of auxin synthetic inhibitors. Collectively, our integrated approaches show that supplying H2 via AB increases LR branching via phytomelatonin signaling. This finding might open the way for applying hydrogen storage materials to horticultural production.

Decoding Microbial Responses to Ammonia Shock Loads in Biogas Reactors through Metagenomics and Metatranscriptomics.

The presence of elevated ammonia levels is widely recognized as a significant contributor to process inhibition in biogas production, posing a common challenge for biogas plant operators. The present study employed a combination of biochemical, genome-centric metagenomic and metatranscriptomic data to investigate the response of the biogas microbiome to two shock loads induced by single pulses of elevated ammonia concentrations (i.e., 1.5 g NH4+/LR and 5 g NH4+/LR). The analysis revealed a microbial community of high complexity consisting of 364 Metagenome Assembled Genomes (MAGs). The hydrogenotrophic pathway was the primary route for methane production during the entire experiment, confirming its efficiency even at high ammonia concentrations. Additionally, metatranscriptomic analysis uncovered a metabolic shift in the methanogens Methanothrix sp. MA6 and Methanosarcina flavescens MX5, which switched their metabolism from the acetoclastic to the CO2 reduction route during the second shock. Furthermore, multiple genes associated with mechanisms for maintaining osmotic balance in the cell were upregulated, emphasizing the critical role of osmoprotection in the rapid response to the presence of ammonia. Finally, this study offers insights into the transcriptional response of an anaerobic digestion community, specifically focusing on the mechanisms involved in recovering from ammonia-induced stress.

Continuous production of high-concentrated ammonia broth through fermentation.

Being considered as a valuable resource and energy carrier, extensive research is going on to efficiently extract ammonia (NH3) from anaerobic digestate. However, due to the well-known NH3 inhibition on methanogens, the total NH3 nitrogen (TAN) concentration is typically limited to 1-4 g N/L in digestate, making the NH3 extraction process energy-consumptive. Here, NH3 fermentation, specifically targeting augmented NH3 production through biological reaction, was performed in a continuous mode. With the increase of gelatin input (10 to 150 g COD/L), NH3 concentration and volumetric productivity gradually increased, reaching 12.0 g TAN-N/L and 36.0 g NH3-N/L/d, which were the highest values ever reported. The stepwise increase in NH3 exposure prompted a shift in microbial dominance towards Hathewaya (from 1 % to 68 %), a critical factor for having high NH3 tolerance. Finally, NH3 stripping results suggested that highly concentrated broth could reduce the specific energy consumption for NH3 extraction to 1/3.

miRNA-ome plasma analysis unveils changes in blood-brain barrier integrity associated with acute liver failure in rats.

BACKGROUND: Hepatic encephalopathy (HE) symptoms associated with liver insufficiency are linked to the neurotoxic effects of ammonia and other toxic metabolites reaching the brain via the blood-brain barrier (BBB), further aggravated by the inflammatory response. Cumulative evidence documents that the non-coding single-stranded RNAs, micro RNAs (miRs) control the BBB functioning. However, miRs' involvement in BBB breakdown in HE is still underexplored. Here, we hypothesized that in rats with acute liver failure (ALF) or rats subjected to hyperammonemia, altered circulating miRs affect BBB composing proteins. METHODS: Transmission electron microscopy was employed to delineate structural alterations of the BBB in rats with ALF (thioacetamide (TAA) intraperitoneal (ip.) administration) or hyperammonemia (ammonium acetate (OA) ip. administration). The BBB permeability was determined with Evans blue dye and sodium fluorescein assay. Plasma MiRs were profiled by Next Generation Sequencing (NGS), followed by in silico analysis. Selected miRs, verified by qRT-PCR, were examined in cultured rat brain endothelial cells. Targeted protein alterations were elucidated with immunofluorescence, western blotting, and, after selected miR mimics transfection, through an in vitro resistance measurement. RESULTS: Changes in BBB structure and increased permeability were observed in the prefrontal cortex of TAA rats but not in the brains of OA rats. The NGS results revealed divergently changed miRNA-ome in the plasma of both rat models. The in silico analysis led to the selection of miR-122-5p and miR-183-5p with their target genes occludin and integrin ß1, respectively, as potential contributors to BBB alterations. Both proteins were reduced in isolated brain vessels and cortical homogenates in TAA rats. We documented in cultured primary brain endothelial cells that ammonia alone and, in combination with TNFα increases the relative expression of NGS-selected miRs with a less pronounced effect of TNFα when added alone. The in vitro study also confirmed miR-122-5p-dependent decrease in occludin and miR-183-5p-related reduction in integrin ß1 expression. CONCLUSION: This work identified, to our knowledge for the first time, potential functional links between alterations in miRs residing in brain endothelium and BBB dysfunction in ALF.

Effects of hydrolyzed chicken liver on digestibility, fecal and urinary characteristics, and fecal metabolites of adult dogs.

High-protein low-carbohydrate diets have been widely used for adult maintenance dogs, as well as in specific weight loss diets and maintenance programs. However, increasing dietary protein may increase undigested protein in the hindgut, modifying intestinal fermentation and fecal metabolite concentrations. The aim of this study was to evaluate the effects of protein source and concentration on apparent total tract digestibility (ATTD) of nutrients, metabolizable energy (ME), fecal and urinary characteristics, and fecal metabolites of dogs. Twelve healthy adult dogs were distributed into six treatments (n = 6 per diet) in a balanced incomplete Latin square design consisting of three periods of 30 days each. Six diets, varying in protein source [poultry byproduct meal (PBPM) and hydrolyzed chicken liver powder (HCLP)] and concentration [24, 32, and 40% crude protein (CP) on dry matter (DM) basis] were tested: PBPM24 (PBPM based diet with 24% CP); PBPM32 (PBPM with 32% CP); PBPM40 (PBPM with 40% CP); HCLP24 (HCLP based diet with 24% CP); HCLP32 (HCLP with 32% CP); HCLP40 (HCLP with 40% CP). The ATTD of CP was greater in dogs-fed HCLP and higher protein concentrations diets (P < 0.05). However, dogs-fed HCLP diets had lower ATTD of fat and carbohydrates, and ME (P < 0.05). Similarly, high-protein diets reduced the ATTD of DM, OM, fat, carbohydrates, and energy (P < 0.05). High-protein diets increased the daily fecal output and moisture (P = 0.004 and P < 0.05, respectively), as well as the fecal score (P < 0.0001), verified as soft, moist stools, but still within the ideal range. Fecal valerate concentration was greater in dogs-fed PBPM at 32% CP (P = 0.007). Fecal isobutyrate tended to increase in dogs-fed PBPM and high-protein diets (P < 0.10). Also, dogs-fed PBPM and high-protein diets had greater fecal concentrations of isovalerate, branched-chain fatty acids (BCFA), and ammonia (P < 0.05). Finally, the fecal lactate concentration increased in dogs-fed HCLP and high-protein diets (P < 0.05). The HCLP increased the ATTD of CP, being a highly digestible protein. Although the inclusion of HCLP slightly increased fecal score and moisture, it decreased the amount of fecal metabolites of protein fermentation ammonia and BCFA, both of which are associated with proteolytic fermentation in the colon.

Feeding companion animals with high-protein diets has been a demand of the market and pet owners. However, the protein quality and quantity consumed can interfere with the amount of undigested protein that reaches the hindgut and be fermented. Intestinal fermentation can be desired when well controlled. This study tested two protein sources (hydrolyzed chicken liver and poultry byproduct meal) combined at three dietary protein concentrations (24, 32, and 40% crude protein on dry matter basis) and their effects on the apparent total tract digestibility (ATTD), fecal and urinary characteristics, and fecal metabolites of healthy adult dogs. In summary, diets containing higher inclusion of hydrolyzed chicken liver had improved protein ATTD. However, the same diets impaired the ATTD of fat and carbohydrates and decreased metabolizable energy. High-protein diets retained more water in the feces and increased the fecal output. Fecal consistency was affected, scored as soft and moist stools, but remained within an acceptable score. Dogs-fed poultry byproduct meal diets had greater concentrations of fecal protein fermentation metabolites, such as ammonia and branched-chain fatty acids, possibly related to a greater amount of undigested protein that reached the hindgut and was fermented.

Anticancer and antibacterial properties of carbon nanotubes are governed by their functional groups.

Due to their high strength, low weight, and biologically-inspired dimensions, carbon nanotubes have found wide interest across all of medicine. In this study, four types of highly dispersible multi-walled carbon nanotubes (CNTs) of similar dimensions, but slightly different chemical compositions, were compared with an unmodified material to verify the impact their surface chemistry has on cytocompatibility, anticancer, inflammation, and antibacterial properties. Minute changes in the chemical composition were found to greatly affect the biological performance of the CNTs. Specifically, the CNTs with a large number of carbon atoms with a +2 coordination number induced cytotoxicity in macrophages and melanoma cells, and had a moderate antibacterial effect against Gram-positive (S. aureus) and Gram-negative (E. coli) bacteria strains, all while being cytocompatible towards human dermal fibroblasts. Moreover, substituting some of the OH groups with ammonia diminished their cytotoxicity towards macrophages while still maintaining the aforementioned positive qualities. At the same time, CNTs with a large number of carbon atoms with a +3 coordination number had a high innate cytocompatibility towards normal healthy cells but were toxic towards cancer cells and bacteria. The latter was further boosted by reacting the CNTs' carboxyl groups with ammonia. Although requiring further analyses, the results of this study, thus, introduce new CNTs that without drugs can treat cancer, inflammation, and/or infection while still remaining cytocompatible with mammalian cells.

Combined effects of dietary carbohydrate levels and ammonia stress on growth, antioxidant capacity and glucose metabolism in juvenile oriental river prawn (Macrobrachium nipponense).

Ammonia is a common environmental stress factor that constrains aquaculture industry development. This study evaluated the effect of carbohydrate levels and ammonia stress in oriental river prawn (Macrobrachium nipponense). The experiment had six treatments containing two water ammonia levels (0 and 5 mg/L) and three dietary carbohydrate levels (low carbohydrate diet (LCD, 10%), medium carbohydrate diet [MCD, 20%], and high carbohydrate diet [HCD, 30%]), and lasted six weeks. The results showed that the prawns fed on MCD had higher weight gain than those fed on LCD and HCD during ammonia stress. Moreover, the prawns fed on MCD had significantly lower acid phosphatase and alkaline phosphatase activities during ammonia stress. Feeding the prawns on the MCD increased B cells in the hepatopancreas during ammonia stress. Interestingly, the prawns fed on MCD had significantly lower superoxide dismutase activity compared to LCD and HCD during ammonia stress. Moreover, the prawns fed on MCD had significantly lower pyruvate kinase activity and pyruvate and lactic acid contents, while those fed on LCD had significantly higher succinic dehydrogenase, 6-phosphogluconic dehydrogenase, and phosphoenol pyruvate carboxykinase activities during ammonia stress. The prawns fed on the MCD increased significantly glutaminase activity and decreased the ammonia content in the serum during ammonia exposure. In addition, feeding the prawns on MCD decreased significantly the expression of apoptosis and inflammation-related genes. Taken together, the MCD supplied energy required to counteract ammonia stress, which increased growth, improved antioxidant capacity, facilitated ammonia excretion, and alleviated inflammation and apoptosis of the oriental river prawn.

Effect of Saengmaeksan on Fatigue, Liver Function, and Immunity Combined with High-Intensity Training.

Saengmaeksan (SMS) is a traditional drink that consists of three natural herbs, Lirio platyphlla, Panax ginseng, and Schisandra chinensis, and improves fatigue, liver function, and immunity. Moderate-intensity exercise has a positive effect on fatigue, liver function, and immune function, whereas long-term high-intensity training has a negative effect on these aspects. We hypothesized that SMS intake will improve fatigue (ammonia, lactic acid), liver function (aspartate transaminidase (AST) and alanine aminotransferase (ALT)), and immunity (IgA, IgG, IgM) with high-intensity training. To investigate this hypothesis, 17 male college tennis players were randomly assigned to SMS and placebo groups with high-intensity training. SMS and placebo were consumed in 110 mL doses for a total of 770 mL. High-intensity training was performed at 70%-90% of heart rate reserve, conducted five times weekly for 4 weeks. A significant interaction effect was observed between the SMS and control (CON) groups regarding ammonia, ALT, and IgA levels. Ammonia levels significantly decreased in the SMS group, but there was no difference in the lactic acid levels. AST significantly decreased in the SMS group. IgA significantly increased in the SMS group, IgM significantly decreased in both groups, but there was no change in IgG. The correlation analysis revealed positive correlation in ΔAST vs. ΔALT, ΔALT vs. ΔIgG, and ΔIgA vs. ΔIgG in the SMS group. These findings demonstrate that SMS intake can reduce ammonia, AST, ALT, and IgM levels, while causing an increase in IgA, which has a positive effect on fatigue reduction, liver function, and immunoglobulins in a high-intensity training or related environment.

An orally deliverable ornithine-based self-assembling polymer nanomedicine ameliorates hyperammonemia in acetaminophen-induced acute liver injury.

l-Ornithine (Orn) is a core amino acid responsible for ammonia detoxification in the body via the hepatic urea cycle. Clinical studies in Orn therapy have focused on interventions for hyperammonemia-associated diseases, such as hepatic encephalopathy (HE), a life-threatening neurological symptom affecting more than 80% of patients with liver cirrhosis. However, its low molecular weight (LMW) causes Orn to diffuse nonspecifically and be rapidly eliminated from the body after oral administration, resulting in unfavorable therapeutic efficacy. Hence, Orn is constantly supplied by intravenous infusion in many clinical settings; however, this treatment inevitably decreases patient compliance and limits its application in long-term management. To improve the performance of Orn, we designed self-assembling polyOrn-based nanoparticles for oral administration through ring-opening polymerization of Orn-N-carboxy anhydride initiated with amino-ended poly(ethylene glycol), followed by acylation of free amino groups in the main chain of the polyOrn segment. The obtained amphiphilic block copolymers, poly(ethylene glycol)-block-polyOrn(acyl) (PEG-block-POrn(acyl)), enabled the formation of stable nanoparticles (NanoOrn(acyl)) in aqueous media. We employed the isobutyryl (iBu) group for acyl derivatization in this study (NanoOrn(iBu)). In the healthy mice, daily oral administration of NanoOrn(iBu) for one week did not induce any abnormalities. In the mice exhibiting acetaminophen (APAP)-induced acute liver injury, oral pretreatment with NanoOrn(iBu) effectively reduced systemic ammonia and transaminases levels compared to the LMW Orn and untreated groups. The results suggest that the application of NanoOrn(iBu) is of significant clinical value with the feasibility of oral delivery and improvement in APAP-induced hepatic pathogenesis. STATEMENT OF SIGNIFICANCE: Liver injury is often accompanied by hyperammonemia, a life-threatening condition characterized by elevated blood ammonia levels. Current clinical treatments for reducing ammonia typically entail the invasive approach of intravenous infusion, involving the administration of l-ornithine (Orn) or a combination of Orn and L-aspartate. This method is employed due to the poor pharmacokinetics associated with these compounds. In our pursuit of enhancing therapy, we have developed an orally administrable nanomedicine based on Orn-based self-assembling nanoparticle (NanoOrn(iBu)), which provides sustained Orn supply to the injured liver. Oral administration of NanoOrn(iBu) to healthy mice did not cause any toxic effects. In a mouse model of acetaminophen-induced acute liver injury, oral administration of NanoOrn(iBu) surpassed Orn in reducing systemic ammonia levels and liver damage, thereby establishing NanoOrn(iBu) as a safe and effective therapeutic option.

Accelerated dissipation, soil microbial toxicity and dispersal of antimicrobial resistance in soils repeatedly exposed to tiamulin, tilmicosin and sulfamethoxazole.

The application of manures leads to the contamination of agricultural soils with veterinary antibiotics (VAs). These might exert toxicity on the soil microbiota and threaten environmental quality, and public health. We obtained mechanistic insights about the impact of three VAs, namely, sulfamethoxazole (SMX), tiamulin (TIA) and tilmicosin (TLM), on the abundance of key soil microbial groups, antibiotic resistance genes (ARGs) and class I integron integrases (intl1). In a microcosm study, we repeatedly treated two soils (differing in pH and VA dissipation capacity) with the studied VAs, either directly or via fortified manure. This application scheme resulted in accelerated dissipation of TIA, but not of SMX, and accumulation of TLM. Potential nitrification rates (PNR), and the abundance of ammonia-oxidizing microorganism (AOM) were reduced by SMX and TIA, but not by TLM. VAs strongly impacted the total prokaryotic and AOM communities, whereas manure addition was the main determinant of the fungal and protist communities. SMX stimulated sulfonamide resistance, while manure stimulated ARGs and horizontal gene transfer. Correlations identified opportunistic pathogens like Clostridia, Burkholderia-Caballeronia-Paraburkholderia, and Nocardioides as potential ARG reservoirs in soil. Our results provide unprecedented evidence about the effects of understudied VAs on soil microbiota and highlight risks posed by VA-contaminated manures. ENVIRONMENTAL IMPLICATION: The dispersal of veterinary antibiotics (VAs) through soil manuring enhances antimicrobial resistance (AMR) development and poses a threat to the environment and the public health. We provide insights about the impact of selected VAs on their: (i) microbially-mediated dissipation in soil; (ii) ecotoxicity on the soil microbial communities; (iii) capacity to stimulate AMR. Our results (i) demonstrate the effects of VAs and their application-mode on the bacterial, fungal, and protistan communities, and on the soil ammonia oxidizers; (ii) describe natural attenuation processes against VA dispersal, (iii) depict potential soil microbial AMR reservoirs, essential for the development of risk assessment strategies.

Predation pressure regulates plant growth promoting (PGP) attributes of bacterial species.

AIM: The present study aimed to investigate the effect of bacterivorous soil protists on plant growth promoting (PGP) attributes of bacterial species and their co-inoculative impact on rice seedling growth. METHODS AND RESULTS: The effect of protists on the PGP attributes of bacteria was tested using standard protocols. The results revealed that the plant-beneficial properties of plant growth promoting bacteria (PGPB) were altered in the presence of various protist species. A significant increase in the production of siderophore units (86.66%), ammonia (34.80 µmol mL-1), and phosphate solubilization index (PSI) (5.6) was observed when Bacillus cereus (Bc) and Pseudomonas fluorescens (Ps) were co-inoculated with unidentified species belonging to the family Kreyellidae (C5). In the case of Enterobacter cloacae co-inoculated with C5 (Kreyellidae), a higher amount of siderophore (51.33%), ammonia (25.18 µmol mL-1), and indole-3-acetic acid (IAA)-like substance (28.59 µg mL-1) production were observed. The biofilm-forming ability of B. cereus is enhanced in the presence of Tetrahymena sp. (C2Bc), unidentified Kreyellidae (C5Bc), and Colpoda elliotti (C12Bc), whereas E. cloacae showed higher biofilm formation in the presence of Tetrahymena sp. alone Although IAA production decreased under predation pressure, a significant increase in shoot length (64.24%) and primary root length (98.18%) in co-inoculative treatments (C12Bc and C5Bc) compared to bacteria alone (25% and 61.50% for shoots and roots, respectively) was observed. The results of enhanced PGP attributes and rice seedlings growth under predation pressure correlated with the enhanced bacterial activity under predation pressure and protist involvement in plant growth development. CONCLUSIONS: Protists may act as regulators of the bacterial activities involved in plant growth promotion and thus enhance plant growth.

Effects of Curcumin on Oxidative Stress and Ferroptosis in Acute Ammonia Stress-Induced Liver Injury in Gibel Carp (Carassius gibelio).

This study investigated the potential role of curcumin (CUR) in preventing oxidative stress and ferroptosis induced by ammonia exposure in gibel carp. Experimental fish (initial weight: 11.22 ± 0.10 g, n = 150) were fed diets supplemented with or without 0.5% CUR for 56 days, followed by a 24 h ammonia (32.5 mg/L) exposure. Liver damages (aspartate aminotransferase (AST), alanine aminotransferase (ALT), adenosine deaminase (ADA), and alkaline phosphatase (ALP)) and oxidative stress enzyme activities (reactive oxygen species (ROS), malondialdehyde (MDA); and the content of antioxidant capacity (T-AOC), superoxide dismutase (SOD), and glutathione peroxidase (GPx)) were induced by ammonia stress. The antioxidant capacity was decreased, as indicated by inhibited gene expression of nuclear factor erythroid 2-related factor 2 (nrf2), heme oxygenase-1 (ho-1), catalase (cat), and sod. Ferroptosis was induced by ammonia stress, as suggested by upregulated mRNA levels of nuclear receptor coactivator 4 (ncoa4), transferrin receptor 1 (tfr1), and iron-responsive element-binding protein 2 (ireb2), and downregulated expression of glutathione peroxidase 4 (gpx4), ferroportin (fpn), and ferritin heavy chain 1 (fth1). In addition, both mRNA and protein levels of ferroptosis markers acyl-CoA synthetase long-chain family member 4 (ACSL4) and prostaglandin-endoperoxide synthase 2 (PTGS2) were upregulated, while cystine/glutamate antiporter (SLC7A11) was downregulated. However, liver injury and ferroptosis in fish induced by ammonia could be attenuated by CUR. Collectively, these findings demonstrate that CUR ameliorates oxidative stress and attenuates ammonia stress-induced ferroptosis. This study provides a new perspective on potential preventive strategies against ammonia stress in gibel carp by dietary CUR.

Abiotic and biotic stress alleviating effects of the medicinal and aromatic plant-derived product on striped catfish Pangasianodonhypophthalmus.

Intensification of aquaculture production leads to abiotic and biotic stresses, which are further induced by the impact of climate change. Thus, it is important to explore a combined strategy to alleviate multiple stresses in fish. In the present investigation, the removal of nitrogenous metabolites from aquaponics water was studied using products derived from five different locally available medicinal and aromatic plants (MAPs) namely banana stem (Musa Accuminata), Aloe vera, Mint (Minata sepicata), Indian beech/Pongameoil-tree/Karanj (Pongamia pinnatum), and Coat Button/Mexican daisy/Dagadipala (Tridax procrumbens) in the batch experiment for 24 h. A reduction of 91-94% in the ammonia level and 75-80% removal of nitrite-N in the treatments with products derived from mint, banana stem, and aloe vera has been observed. Mint product was also found to be bactericidal against fish pathogenic bacteria. Based on this dual function of mint, further study was conducted under wet-lab conditions to evaluate the possible protective role of the mint-based product in dry powder form against abiotic stresses caused by nitrogenous toxicants for 24 h and 45 days multiple stresses caused by nitrogenous toxicants in freshwater stocked with eight fingerlings of Pangasianodon hypophthalmus of the initial weight of 8 ± 0.5g for 24 h and 8.82 ± 0.75g for 45 days. The results revealed that mint-derived plant product has better anti-stress properties in terms of their bioremediation effectiveness in lowering ammonia and nitrite. Mint also improved the fish growth performance with better physiological responses and anti-oxidative status and reduced the cellular metabolic stress in fish reared under ammonia, as further indicated by reduced oxidative stress parameters, and tissue biochemical indices. Overall, mint showed its main properties as a growth promoter, and immune enhancer, where this medicinal plant product acts as a bioremediation and antibacterial agent to the host immune system. This has potential applications in the environmental and health management of aquaculture.

Nematode-Suppressive Potential of Digestates to Meloidogyne incognita and Heterodera schachtii.

Management of plant-parasitic nematodes uses host plant resistance, crop rotation, cultural methods, and nematicide applications. Host plant resistance is tedious to develop, and crop rotation and cultural methods are challenging to use. Environmental and human health concerns render sole reliance on chemical nematode suppression nonsustainable. Previously, digestate from anaerobically fermented maize silage suppressed Heterodera schachtii in Beta vulgaris crops. Here, seven digestates were investigated for nematode suppressive potential: liquid dairy manure digestate (LDMD), liquid dairy manure digestate with ammonia removed (LDMDA-), food waste digestate (FWD), liquid food waste digestate with ammonia removed (LFWDA-), liquid food waste digestate (LFWD), food waste hydrolysate from the Renewable Energy Anaerobic Digester (HREAD), and food waste hydrolysate from the South Area Transfer Station in Sacramento (HSATS). In a red radish (Raphanus sativus) bioassay with H. schachtii, digestates were amended at rates of 0.02, 0.11, 0.57, and 2.86 ml per 100 cm3 of soil. At a rate of 2.86 ml, all amendments except LDMDA- and LFWDA- significantly reduced juvenile root penetration compared with the infested control. In a greenhouse watermelon (Citrullus lanatus) bioassay with Meloidogyne incognita, amendments FWD, LFWD, HREAD, and HSATS as well as LDMD (less effectively) at 2.86 and 5.76 ml per 100 cm3 of soil significantly reduced egg masses per root system compared with the nontreated, nematode-infested control. In a microplot experiment with M. incognita and red radish, in the treatment amended with LFWD at 2.37 ml per 100 cm3 of soil, marketable yields were improved by approximately 50% over the nontreated control and were comparable with those in the treatment with the nematicide Reklemel. In a second microplot experiment with M. incognita and watermelon, treatments that contained LFWD at rates of 3.55 ml per 100 cm3 of soil had transient numerical effects of initial nematode suppression that were not maintained throughout the 3-month growth period. The results of these studies demonstrated that digestates FWD and LFWD consistently expressed some nematode-suppressive capacity.

Growth-promoting and immunostimulatory effects of phytobiotics as dietary supplements for Pangasianodon hypophthalmus fingerlings.

A 56-day feeding experiment was designed to evaluate the impacts of five herbal extracts, namely Ginkgo biloba (GB), Moringa oleifera (MR), Myristica fragrans (NM), Silybum marianum (MT), and Astragalus membranaceus (AT) on growth, serum immune indices, and ammonia-N stress resistance of Pangasianodon hypophthalmus fingerlings. Fish were fed on a diet with no herbal additives (reference or CNT), and several plant extracts-based diets were supplied with two inclusion doses of each extract (1.0 and 2.0 g/kg). After the end of the feeding, fish in all groups were exposed to acute ammonia stress, mortalities were recorded every 2 h for a duration of 10 h, and then survival percent was assessed. Results showed that growth was upgraded significantly in groups fed plant extracts-based diets compared to the CNT group. Serum proteins (albumin and total protein), lysozyme, complement C3, bactericidal, and myeloperoxidase activities were increased significantly in groups fed plant extracts-based diets in relation to the CNT group. The total immunoglobulin levels were increased significantly only in fish groups fed GB, MT, and NM-based diets. Post-exposure to acute ammonia stress, the Kaplan-Meier survival curve presented significantly higher survival of fish groups that fed plant extracts-enriched diets than those fed the CNT diet. The aforementioned results suggest that using herbal extracts as feed supplements can beneficially enhance the growth, the immunity of P. hypophthalmus fingerlings and may increase their tolerance in the face of extrinsic stressors. These findings may pave the way for the potential and regular application of herbal extracts in diets of P. hypophthalmus at their early life stages to raise their immunity and maintain aquaculture sustainability.

Gasotransmitter ammonia accelerates seed germination, seedling growth, and thermotolerance acquirement in maize.

Ammonia (NH3), as an intermediate product of nitrogen metabolism, is recognized as a novel gasotransmitter (namely gaseous signaling molecule), its signaling role being revealed in plants. NH3 exists in two different chemical forms, namely the weak base (free molecule: NH3) and the weak acid (ammonium: NH4+), which are generally in equilibrium with each other in plants. However, the effect of NH3 on seed germination, seedling growth, and thermotolerance acquirement in maize remains unclear. Here, maize seeds were imbibed in the different concentrations of NH3·H2O (NH3 donor), and then germinated and calculated seed germination rate at the various time points. Also, the 60-h-old seedlings were irrigated in the different concentrations of NH3·H2O, and then subjected to heat stress and counted survival rate. The data implied that the appropriate concentrations (6, 9, and 12 mM) of NH3·H2O accelerated seed germination as well as increased seedling height and root length compared with the control without NH3 treatment. Also, the suitable concentrations (2 and 4 mM) of NH3·H2O improved tissue vitality, relieved an increase in malondialdehyde content, and enhanced survival rate of maize seedlings under heat stress compared with the control. These results firstly suggest that NH3 could accelerate seed germination, seedling growth, and thermotolerance acquirement in maize.

Modified dietary fiber from cassava pulp affects the cecal microbial population, short-chain fatty acid, and ammonia production in broiler chickens.

The objective of this study was to investigate the effects of modified dietary fiber from cassava pulp (M-DFCP) supplementation in broiler diets on cecal microbial populations, short-chain fatty acids (SCFAs), ammonia production, and immune responses. A total of 336, one-day-old male broiler chicks (Ross 308) were distributed over 4 dietary treatments in 7 replicate pens (n = 12 chicks) using a completely randomized design. Chicks were fed the control diet and 3 levels of M-DFCP (0.5, 1.0, and 1.5%) for an experimental duration of 42 d. The M-DFCP contained total dietary fiber (TDF), soluble dietary fiber (SDF), insoluble dietary fiber (IDF), cello-oligosaccharides (COS), and xylo-oligosaccharides (XOS) of approximately 280.70, 22.20, 258.50, 23.93, and 157.55 g/kg, respectively. The 1.0 and 1.5% M-DFCP supplementation diets showed positive effects on stimulating the growth of Lactobacillus spp. and Bifidobacterium spp., enhancing SCFAs (acetic, propionic, butyric acid, and branched SCFAs) and lactic acid concentrations during growing periods. Broilers fed 1.0 and 1.5% M-DFCP also exhibited a significant increase in caecal Lactobacillus spp. and lactic acid concentrations during the finisher period as well. In addition, M-DFCP also reduced cecal digesta and excreta ammonia production in broilers over both periods (0-21 and 22-42 d of age). However, M-DFCP did not exhibit any effect on total serum immunoglobulin (Ig) or lysozyme activity. In conclusion, this study shows that M-DFCP can be used as a dietary fiber source in broiler diets, with a recommended level of approximately 1.0%.