Dietary manganese nano-particles improves gene regulation and biochemical attributes for mitigation of lead and ammonia toxicity in fish.

In the present study, we explored the capability of manganese nanoparticles (Mn-NPs) to alleviate the toxicity induced by lead (Pb) and ammonia (NH3) toxicity in Oreochromis niloticus (GIFT strain). The experiment followed a completely randomized design, including a control group (Mn-NPs-0 mg kg-1 diet) and groups exposed to Pb and NH3 alongwith Mn-NPs at 2 and 3 mg kg-1. Cortisol levels were significantly elevated in Pb + NH3 group whereas reduced by Mn-NPs diets. Gene expressions of HSP 70, iNOS, CYP 450, and Cas 3a were notably upregulated by Pb + NH3 group and downregulated by Mn-NPs diets. The cellular metabolic enzymes were affected by Pb + NH3 exposure and mitigated by Mn-NPs diets. The liver and kidney exhibited reduced activities of catalase, superoxide dismutase, and glutathione-s-transferase with Mn-NPs diets. Concurrently, immune-related genes such as total immunoglobulin (Ig) and tumor necrosis factor (TNFα) were upregulated in the Mn-NPs-fed groups. Growth performance indicators, including weight gain %, feed conversion ratio, specific growth rate, protein efficiency ratio, and relative feed intake were adversely affected by Pb + NH3 stress but improvement with Mn-NPs diets. Genes associated with growth performance, such as growth hormone (GH), growth hormone regulatory (GHR1), and myostatin, exhibited enhancements in response to Mn-NPs diets. Digestive enzymes, including protease and amylase were also enhanced by Mn-NPs diets. Additionally, Mn-NPs diets led to a reduction in the bioaccumulation of lead. This study aims to investigate the role of Mn-NPs in mitigating the effects of lead and ammonia toxicity on fish by examining various biochemical and gene regulatory factors to enhance fish wellbeing.

Combined effects of water salinity and ammonia exposure on the antioxidative status, serum biochemistry, and immunity of Nile tilapia (Oreochromis niloticus).

Growing Nile tilapia in brackish water showed promising results, but the possibility of ammonia exposure can interrupt health status and productivity. Herein, the study tested the combined effects of water salinity and ammonia exposure on the antioxidative status, serum biochemistry, and immunity of Nile tilapia. Fish were assigned to eight groups where fish were reared in saline water (5, 10, and 15 ppt) with continuous or intermittent (every 3 days) total ammonia (TAN) exposure (5 mg TAN/L) (2 × 4 factorial design). After 30 days, the water salinity, TAN, and their interaction were markedly (P < 0.05), affecting the growth performance (final weight, weight gain, and specific growth rate) and survival rate of Nile tilapia. The growth performance and survival rate were markedly lower in tilapia grown in 15 ppt with continuous TAN exposure than in the remaining groups. The results showed that fish exposed to higher salinity levels (10 and 15 ppt) and continuous TAN exposure had a more robust antioxidative response, as evidenced by higher superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) activities and lower malondialdehyde (MDA) levels in the homogenates of the gills, intestines, and livers. The gills were notably affected, with congestion of primary filaments blood vessels and degeneration or shedding of secondary filaments epithelium, especially at salinity levels of 10 and 15 ppt. Additionally, the intestines displayed hyperplasia and inflammatory cell infiltration of intestinal mucosa at 5-10 ppt salinity, degeneration and sloughing of the intestinal epithelium at 15 ppt saline water, and increased goblet cell number at salinity of 10 ppt. The study found that continuous TAN exposure had a more significant impact on the fish, especially at higher salinity levels. Water salinity, TAN, and their interaction significantly affected all measured blood bio-indicators (total, albumin, globulin, ALT, AST, creatinine, urea, glucose, and cortisol levels). The phagocytic activity and index were markedly lowered in fish reared in 15 ppt with continuous TAN exposure, while the lysozyme activity was decreased in fish grown in 5, 10, and 15 ppt with continuous TAN exposure. In conclusion, Nile tilapia showed the possibility of growth with normal health status in brackish water (5-10 ppt); however, continuous TAN exposure can impair the productivity of tilapia, especially with high salinity (15 ppt).

α-lipoic acid ameliorates hepatotoxicity induced by chronic ammonia toxicity in crucian carp (Carassius auratus gibelio) by alleviating oxidative stress, inflammation and inhibiting ERS pathway.

High environment ammonia (HEA) poses a deadly threat to aquatic animals and indirectly impacts human healthy life, while nutritional regulation can alleviate chronic ammonia toxicity. α-lipoic acid exhibits antioxidative effects in both aqueous and lipid environments, mitigating cellular and tissue damage caused by oxidative stress by aiding in the neutralization of free radicals (reactive oxygen species). Hence, investigating its potential as an effective antioxidant and its protective mechanisms against chronic ammonia stress in crucian carp is highly valuable. Experimental fish (initial weight 20.47 ± 1.68 g) were fed diets supplemented with or without 0.1% α-lipoic acid followed by a chronic ammonia exposure (10 mg/L) for 42 days. The results revealed that chronic ammonia stress affected growth (weight gain rate, specific growth rate, and feed conversion rate), leading to oxidative stress (decreased the activities of antioxidant enzymes catalase, superoxide dismutase, glutathione peroxidase; decreased total antioxidant capacity), increased lipid peroxidation (accumulation of malondialdehyde), immune suppression (decreased contents of nonspecific immune enzymes AKP and ACP, 50% hemolytic complement, and decrease of immunoglobulin M), impaired ammonia metabolism (reduced contents of Glu, GS, GSH, and Gln), imbalance of expression of induced antioxidant-related genes (downregulation of Cu/Zu SOD, CAT, Nrf2, and HO-1; upregulation of GST and Keap1), induction of pro-apoptotic molecules (transcription of BAX, Caspase3, and Caspase9), downregulation of anti-apoptotic gene Bcl-2 expression, and induction of endoplasmic reticulum stress (upregulation of IRE1, PERK, and ATF6 expression). The results suggested that the supplementation of α-lipoic acid could effectively induce humoral immunity, alleviate oxidative stress injury and endoplasmic reticulum stress, and ultimately alleviate liver injury induced by ammonia poisoning (50-60% reduction). This provides theoretical basis for revealing the toxicity of long-term ammonia stress and provides new insights into the anti-ammonia toxicity mechanism of α-lipoic acid.

Effects of dietary Plantago ovata seed extract administration on growth performance and immune function of common carp (Cyprinus carpio) fingerling exposed to ammonia toxicity.

Medicinal plants are powerful antioxidants which can improve well-being and suppress oxidative stress caused by environmental toxins in aquatic animals. In this regard, the present research was designed to show the potential effects of psyllium (Plantago ovata) seed extract (PSE) on the growth, and immune responses of common carp Cyprinus carpio exposed to acute ammonia toxicity. To perform the study, fish were fed with diets containing 0 (T0), 0.25 (T1), 0.5 (T2), and 1% (T3) PSE for 60 days, and then exposed to ammonia (0.5 mg L-1) for 3 h. The findings showed that fish given the T1 diet outperformed the T3 and control groups in terms of ultimate weight, weight increase, and food conversion ratio. Additionally, the T1 group showed a significantly higher level of total protein and serum lysozyme activity than the other treatment groups. Moreover, the highest serum total immunoglobulin values were recorded in T1 and T2 groups. The results showed that PSE, especially at moderate levels, could successfully upregulate the transcription of immune-related genes (IFN-γ, Hsp70, TNF-ɑ, IL-1ß, IL-10, and IgE) compared to the control group after exposure to ammonia. Furthermore, improving ammonia-induced down regulations of antioxidant-related gene expressions (CYP1A, SOD, and GPX) was observed in fish fed with PSE-included diets compared to the control one. However, PSE-supplemented diets did not affect the mRNA expression level of CAT. Regarding tight junction-associated genes, the higher mRNA expression level of occludin was observed in the T1 group, whereas the downregulation of CLD3 gene occurred in all experimental groups. Conversely, significant upregulation of osmoregulation-associated gene (NKA) was recorded in all experimental groups compared to the control one. Therefore, the administration of PSE (0.25% of the diet) for 60 days is recommended to increase growth performance, improve health, and increase the resistance of common carp to oxidative stress caused by ammonia.

Selenium Alleviates Ammonia-Induced Splenic Cell Apoptosis and Inflammation by Regulating the Interleukin Family/Death Receptor Axis and Nrf2 Signaling Pathway.

Ammonia (NH3) is a harmful gas in livestock houses. So far, many researchers have demonstrated that NH3 is detrimental to animal and human organs. Selenium (Se) is one of the essential trace elements in the body and has a good antioxidant effect. However, there was little conclusive evidence that Se alleviated NH3 poisoning. To investigate the toxic mechanism of NH3 on pig spleen and the antagonistic effect of L-selenomethionine, a porcine NH3-poisoning model and an L-selenomethionine intervention model were established in this study. Our results showed that NH3 exposure increased the apoptosis rate, while L-selenomethionine supplementation alleviated the process of excessive apoptosis. Immunofluorescence staining, real-time quantitative polymerase chain reaction (qRT-PCR), and western blot results confirmed that exposure to NH3 changed the expression levels of interleukin family factors, apoptosis, death receptor, and oxidative stress factors. Our study further confirmed that excessive NH3 induced inflammatory response and mediated necroptosis leading to cell apoptosis by activating the Nrf2 signaling pathway. Excessive NH3 could mediate spleen injury through oxidative stress-induced mitochondrial dynamics disorder. L-Selenomethionine could alleviate inflammation and abnormal apoptosis by inhibiting the IL-17/TNF-α/FADD axis. Our study would pave the way for comparative medicine and environmental toxicology.

Intestinal barrier dysfunction induced by ammonia exposure in pigs in vivo and in vitro: The protective role of L-selenomethionine.

Ammonia has been reported to have a variety of toxicity to aquatic animals, farm animals and humans. However, its potential toxicity on the intestines remains unknown. L-selenomethionine is one of the important organic selenium sources. However, the mitigating effect of L-selenomethionine on ammonia exposure toxicity is still lacking. Therefore, in this study, the mechanism of toxic action of ammonia on intestinal tract and the detoxification effect of L-selenomethionine were examined. We evaluated the intestinal toxicity of ammonia and the alleviating effect of L-selenomethionine in an in vivo model, and then verified it in vitro model by a variety of cutting-edge experimental techniques. Our results showed that ammonia exposure causes oxidative stress, necroptosis, Th1/Th2 imbalance and inflammation in the intestinal tissue and the intestinal cells, and L-selenomethionine had a significant mitigation effect on the changes of these indexes induced by ammonia. In conclusion, ammonia exposure caused oxidative stress and Th1/Th2 imbalance in the porcine small intestine and IPEC-J2 cells, and that excessive ROS accumulation-mediated necroptosis targeted inflammatory responses, resulting in the destruction of tight connections of intestinal cells, thereby causing intestinal barrier dysfunction. L-selenomethionine could effectively reduce the intestinal injury caused by ammonia exposure and antagonize the toxic effect of ammonia.

The use of selenomethionine to reduce ammonia toxicity in porcine spleen by inhibiting endoplasmic reticulum stress and autophagy mediated by oxidative stress.

Ammonia (NH3) is a typical pollutant in the atmosphere and is well known for its harmful effects on plants, animals as well as human health. Previous studies have shown that NH3 exposure can cause damage to immune organs and impaired immune function in animals. Selenomethionine is a kind of organic selenium, which can not only promote the growth and development of the body, but also inhibit the generation of intracellular reactive oxygen species (ROS), and effectively improve the immune function of the body. Therefore, this study evaluated the toxic effect of NH3 exposure on spleen from a new perspective and investigated the protective effect of selenomethionine on ammonia-induced immunotoxicity. Twenty-four Large White*Duroc*Min pigs were randomly assigned to 4 groups: control group, NH3 group, selenium group, and NH3 + selenium group. Our results showed that NH3 inhalation caused autophagy in the pig spleen, a decrease in lymphocytes, and an increase in autophagic vesicles. Also, NH3 exposure led to a decrease in the activity of some antioxidant enzymes (decreased by about 50%) and a significant increase in the expression of genes related to oxidative stress and endoplasmic reticulum stress (ERS). Our results indicated that selenomethionine mitigated ammonia toxicity in pigs (alleviated about 20-55%). In summary, our findings should be of value in providing a theoretical basis for revealing the toxicity of the high-risk gas NH3, and providing a new perspective on the mechanism of Se against toxic substances.

Protective effects of dietary supplementation of probiotic Bacillus licheniformis Dahb1 against ammonia induced immunotoxicity and oxidative stress in Oreochromis mossambicus.

The goal of this study was to assess the efficacy of probiotics in mitigating ammonia-induced toxicity in fish. Fish were divided into four groups: control, only probiotic, only ammonia, and combined ammonia + probiotic. For 8 weeks, the Oreochromis mossambicus were exposed to waterborne ammonia at 1.0 mg L-1 and/or dietary Bacillus licheniformis Dahb1 at 107 cfu g-1. After the 4th and 8th weeks, the fish were evaluated for growth performance, enzymatic and non-enzymatic antioxidant activities (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) reduced glutathione (GSH), neurotoxicity (acetylcholinesterase - AChE), non-specific immune responses (lysozyme (LYZ), myeloperoxidase (MPO), reactive nitrogen and oxygen species (RNS and ROS) and oxidative stress effects (lipid peroxidation (LPO), DNA damage)). Our results showed that in the absence of waterborne ammonia exposure, B. licheniformis Dahb1 significantly improved growth performance, enzymatic and non-enzymatic antioxidant capacity, AChE activity, non-specific immune response and decreased oxidative stress effect. Ammonia exposure resulted in significantly lower growth performance, reduced enzymatic and non-enzymatic antioxidant ability, decreased AChE activity, decreased non-specific immune response and increased oxidative stress effect. When O. mossambicus were exposed to ammonia, supplementation with B. licheniformis Dahb1 in the diet significantly increased survival, indicating that it may have a significant protective effect against ammonia toxicity by enhancing enzymatic and non-enzymatic antioxidant ability, activity of AChE, non-specific immune response and reduced oxidative stress effect. According to our findings, diet supplementation of B. licheniformis Dahb1 (107 cfu g-1) has the potential to combat ammonia toxicity in O. mossambicus.

Evaluation of potential cardiotoxicity of ammonia: l-selenomethionine inhibits ammonia-induced cardiac autophagy by activating the PI3K/AKT/mTOR signaling pathway.

Ammonia is a major harmful gas in the environment of livestock and poultry. Studies have shown that excessive ammonia inhalation has adverse effects in pig heart. However, the mechanism of ammonia-induced cardiac toxicity in pigs has not been reported. L-selenomethionine is a kind of organic selenium (Se) which is easily absorbed by the body. Therefore, in this study, twenty-four 125-day-old pigs were randomly divided into 4 groups: C (control) group, A (ammonia) group, Se group (Se content: 0.5 mg kg-1), and A (ammonia) + Se group. The mechanism of ammonia-induced cardiotoxicity and the alleviating effect of L-selenomethionine were examined. The results in the A group showed as follows: a large number of myocardial fiber edema and cytoplasmic bleakness were observed in the heart; a large number of mitochondrial autophagy were observed; ATP content, ATPase activities and hematological parameters decreased significantly; Endoplasmic reticulum stress (ERS) markers (GRP78, IRE1α, ATF4, ATF6, and CHOP) were significantly induced in the mRNA and protein levels; PI3K/AKT/mTOR signaling pathway was activated; and autophagy key genes and proteins (Beclin-1, LC3, ATG3, and ATG5) were significantly up-regulated. The results of comparison between the A + Se group and the A group were as follows: the degree of edema of cardiac muscle fiber in the A + Se group was somewhat relieved; the level of mitochondrial autophagy decreased; ATP content and ATPase activities increased significantly; the mRNA and protein levels of ERS markers were significantly down-regulated; the expression level of PI3K/AKT/mTOR signaling pathway was decreased; and the mRNA and protein levels of key autophagy genes were decreased. However, the changes of these indexes in the A + Se group were still significantly different from those in the C group. Our results indicated that L-selenomethionine supplementation inhibited ammonia-induced cardiac autophagy by activating the PI3K/AKT/mTOR signaling pathway, which confirmed that L-selenomethionine could alleviate the cardiac injury caused by excessive ammonia inhalation to a certain extent. This study aims to enrich the toxicological mechanism of ammonia and provide valuable reference for future intervention of ammonia toxicity.

Effects of ammonia toxicity on the histopathology, detoxification, oxidative stress, and immune response of the cuttlefish Sepia pharaonis and the mitigation of γ-aminobutyric acid.

In this study, γ-aminobutyric acid (GABA) was examined as an additional supplement to improve the ammonia stress resistance of S. pharaonis. Specifically, we added different doses of GABA (0, 20, 40, 60, 80, and 100 mg/kg) to food, cultivated S. pharaonis in regular seawater for 8 weeks and then in 8.40 mg/L ammonia seawater for 48 h and then investigated the accumulation of ammonia (the hepatic ammonia content), ammonia detoxification process (the urea content), antioxidant enzymes (superoxide dismutase (SOD) and catalase (CAT) enzyme activities), immune response (the serum haemolytic complement (C3) and lysozyme (LYZ) contents), membrane lipid peroxidation (malondialdehyde (MDA)) and histopathology of the liver. The results showed that ammonia poisoning could induce ammonia and MDA accumulation and subsequently lead to oxidative stress (decreases in SOD and CAT activities), immunosuppression (reductions in the haemolytic C3 and LYZ content), and histopathological injury in the liver. The application of GABA had a significant effect on alleviating the adverse effect of ammonia poisoning, and 80-100 mg/kg treatment exerted the best effect. This treatment significantly reduced the ammonia and MDA contents, significantly increased the urea content, increased the SOD, CAT, C3 and LYZ activities, reduced the MDA content, suppressed membrane lipid peroxidation, and significantly improved the histopathological injury to the liver. In summary, the results could provide a new method for mitigating liver damage, alleviating the physiological and metabolic disorders caused by ammonia stress in cuttlefish, and provide a theoretical basis for the application of GABA in alleviating ammonia poisoning.

Evaluation of L-Selenomethionine on Ameliorating Cardiac Injury Induced by Environmental Ammonia.

L-Selenomethionine is one of the important organic selenium sources. The supplementation of L-selenomethionine in diets is significant to improve the health of pigs. Ammonia is a major pollutant in the atmosphere and piggery, posing a threat to human and animal health. Although ammonia exposure can damage the heart, the mechanism of cardiac toxicity by ammonia is still unknown. In this study, we investigated the mechanism of cardiac injury induced by ammonia exposure in pigs and the protective effect of L-selenomethionine on its cardiotoxicity. The results showed that the blood ammonia content of pig increased significantly in ammonia group, the expressions of energy metabolism-related genes (LDHA, PDK4, HK2, and CPTIB) and the oxidative stress indexes were significantly changed (P < 0.05), the AMPK/PPAR-γ/NF-κB signaling pathways were activated, the chromatin edge aggregation and nuclear pyknosis were observed in ultrastructure, the apoptotic cells were significantly increased (P < 0.05), and the mRNA and protein expressions of apoptosis-related genes (Bcl-2, Bax, Cyt-c, caspase-3, and caspase-9) were significantly affected (P < 0.05). The above changes were significantly alleviated in ammonia + L-selenomethionine group, but there were still significant differences compared with the C group (P < 0.05). Our results indicated that ammonia exposure could cause energy metabolism disorder and oxidative stress and induce apoptosis of cardiomyocytes through AMPK/PPAR-γ/NF-κB pathways, which could lead to cardiac injury and affect cardiac function. L-Selenomethionine could effectively alleviate the cardiac damage caused by ammonia and antagonize the cardiotoxicity of ammonia.

Chemical profiling of Fritillariae thunbergii Miq prepared by different processing methods reveals two new quality markers: Zhebeininoside and imperialine-3-ß-D-glucoside.

ETHNOPHARMACOLOGICAL RELEVANCE: Fritillariae thunbergii Miq (FTM)exhibit versatile biological activities including the significant antitussive and expectorant activities. As a herbal medicine, the therapeutic effects of FTM may be expressed by multi-components which have complicated integration effects on multi-targets. With the time going, the different processing methods of FTM has been changed a lot. Thus，the study described the effect of processing methods to FTM and its quality. MATERIAL AND METHOD: Studies were undertaken by using UHPLC-LTQ Orbitrap MS and pharmacodynamic models. All reagents were involved of analytical grade. While a HPLC-ELSD's method has been developed and validated, a certified Quality System is conformed to ICH requirements. The experimental animals followed the animal welfare guidelines. AIM OF THE STUDY: We aimed to found the differences after the different processing methods of FTM, and to demonstrate the changes could be selected as quality control indicators, and established a method for simultaneous determination of these for quality control. RESULTS: we have previously found two new steroidal alkaloids: zhebeininoside and imperialine-3-ß-D-glucoside from the different processing methods of FTM, which is the difference between the different processing methods of FTM, mainly on the steroidal alkaloids. The activity analysis of zhebeininoside, imperialine-3-ß-D-glucoside, verticine and verticinone showed that the mouse model of cough expectorant has antitussive effect. The positive drug selected was dextromethorphan syrup. The positive group showed biological activity, but the blank group showed nothing. The model group showed illness which means that the model was effective. There are two ways of the mechanism of action of the expectorant action which can make sputum thin, reduce its viscosity, and be easy to cough up, or can accelerate the movement of mucous cilia in the respiratory tract and promote the discharge of sputum. In our study, the content of phenol red was significantly reduced in the administration group. CONCLUSIONS: To sum up, our results suggest that zhebeininoside and other three components cloud be selected as quality control indicators, and a method for simultaneous determination of zhebeininoside and other three components was established for quality control.

Effects of dietary tea tree oil on the growth, physiological and non-specific immunity response in the giant freshwater prawn (Macrobrachium rosenbergii) under high ammonia stress.

This study aimed to investigate the effects of dietary tea tree oil (TTO) on the performance, intestinal antioxidant capacity, and non-specific immunity after ammonia nitrogen stress in Macrobrachium rosenbergii. Six experimental diets were formulated with 0, 25, 50, 100, 200, 400 mg/kg TTO, respectively. A total of 900 prawns (average initial weight, 0.39 ± 0.01 g) were randomly assigned to 6 groups in triplicate in 18 tanks. After an 8-week feeding trial, 20 prawns from each tank were changed with 20 mg/L ammonia stress for 24 h. The results showed that 100 mg/kg TTO significantly increased prawns performance and survival rate compared with the control group. Moreover, 100 and 200 mg/kg TTO significantly improved intestinal antioxidant capabilities by increasing SOD enzyme activities and decreasing MDA levels. In addition, the prawns fed with 100 mg/kg TTO diet showed the highest survival rate under ammonia stress. After ammonia stress, the group of 100 mg/kg TTO significantly improved antioxidant capacity by increasing hemolymph respiratory burst activity, as well as intestinal anti-superoxide anion activity and SOD. Coincidentally, 100 mg/kg TTO significantly upregulated the intestinal relative expression of antioxidant-related genes (peroxiredoxin-5). Further, it was found that 100 mg/kg TTO activated the toll-dorsal pathway in prawns, which performed the similar function as the classic NF-κB pathway by upregulating the TNF-α and IL-1. Finally, 100 mg/kg TTO increased the levels of iNOS activities and NO contents after ammonia stress and enhanced non-specific immunity. The results indicated that 100 mg/kg TTO could significantly improve the M. rosenbergii performance, antioxidant capacity and ammonia stress resistance. We suggested that the mechanisms may be attributed to that TTO enhanced the antioxidant capacity and non-specific immunity of M. rosenbergii via the NF-κB signal pathway.

Effects of dietary inclusion of Radix Bupleuri extract on the growth performance, and ultrastructural changes and apoptosis of lung epithelial cells in broilers exposed to atmospheric ammonia.

To explore whether Radix Bupleuri extract (RBE) could protect lung injury of broilers under ammonia (NH3) exposure, 360 one-d-old male broilers were randomly allocated to four groups of six replicates each in a 2 × 2 factorial design with two diets (the basal diet [control; CON] and the basal diet supplemented with RBE [RB]) and two air conditions (normal condition [<2 ppm of NH3; NOR] and NH3 exposure [70 ppm of NH3; NH70]). The RB diet contained 80 mg saikosaponins/kg diet. On day 7, the lung tissues were collected and the lung epithelial cells (LEC) were isolated. Our experimental results showed that the NH3 exposure decreased body weight gain and feed intake irrespective of dietary treatments during days 1 to 7. However, the RBE addition decreased feed consumption to body weight gain ratio in broilers under NH70 conditions. In the LEC of CON-fed broilers under NH70 conditions, Golgi stacks showed the dilation of cisternaes and reduced secretory vesicles, mitochondria enlarged, the inner membrane of mitochondria became obscure, and the cristae of mitochondria ruptured, whereas only a mild enlargement of Golgi cisternaes and the part rupture of mitochondrial cristaes occurred in the LEC of RB-fed broilers under NH70 conditions. The NH3 exposure increased malondialdehyde (MDA) level, but decreased total antioxidant capacity (T-AOC) in the lungs of CON-fed broilers. However, the RBE addition decreased MDA level and increased T-AOC in the lungs of broilers under NH70 conditions. Simultaneously, the NH3 exposure increased apoptotic rate (AR), mitochondrial membrane potential (MMP), and reactive oxygen species (ROS) level in the isolated LEC of CON-fed broilers. The RBE addition decreased AR, MMP, and ROS in the isolated LEC of broilers under NH70 condition. Besides, the NH3 exposure increased mRNA expression of B-cell lymphoma-2 associated X protein (BAX), caspase-3, and tumor necrosis factor α (TNF-α), but increased interferon γ (IFN-γ) mRNA abundance in the lungs of CON-fed broilers. The RBE supplement decreased mRNA levels of BAX, caspase-3, and TNF-α, but increased IFN-γ, interleukin (IL)-4, and IL-17 mRNA levels in the lungs of broilers under NH70 conditions. These results indicated that dietary RBE addition alleviated NH3 exposure-induced intercellular ultrastructural damage via mitochondrial apoptotic pathway, possibly due to RBE-induced increase of antioxidant capacity and immunomodulatory function in the lungs of broilers under NH3 exposure.

Gill oxidative damage caused by acute ammonia stress was reduced through the HIF-1α/NF-κb signaling pathway in golden pompano (Trachinotus ovatus).

This study aimed to investigate the intoxication mechanism of golden pompano (Trachinotus ovatus) exposed to high ammonia levels and the effects on the immune and antioxidant mechanisms of gills. Juvenile golden pompano was exposed to ammonia (total ammonia: 26.9 mg/L) to induce 96 h of ammonia stress, and a 96 h recovery experiment was performed after poisoning. Then, we evaluated hematological parameters, the histological structure and the expression of related genes. In this experiment, continuous exposure to high levels of ammonia led to a significant increase in plasma alkaline phosphatase (ALP), acid phosphatase (ACP) and lactate dehydrogenase (LDH) levels (P < 0.05), and the levels of triiodothyronine (T3) and tetraiodothyronine (T4) were significantly reduced (P < 0.05). Moreover, the expression of antioxidant genes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and inflammatory cytokines such as tumor necrosis factor α (TNF-α) and interleukin 1ß (IL-1ß) increased (P < 0.05). These results indicate that ammonia activates the active osmotic regulatory mechanism of fish gills and participates in defense and immune responses. However, with prolonged exposure to ammonia, the balance of the defense system is disrupted, leading to oxidative damage and inflammation of the gill tissue. This research not only helps elucidate the intoxication mechanism of golden pompano by ammonia at the molecular level but also provides a theoretical basis for further research on detoxification mechanisms.

Ammonia exposure causes the disruption of the solute carrier family gene network in pigs.

Ammonia is the main harmful gas in livestock houses. However, the toxic mechanism of ammonia is still unclear. Therefore, we examined the effects of ammonia exposure on different tissues of fattening pigs by histological analysis and transcriptome techniques in this study. The results showed that there were varying degrees of pathological changes in liver, kidney, hypothalamus, jejunum, lungs, spleen, heart and trachea of fattening pigs under ammonia exposure. Notably, the extent of damage in liver, kidney, jejunum, lungs, hypothalamus and trachea was more severe than that in heart and spleen. Transcriptome results showed that ammonia exposure caused changes in 349, 335, 340, 229, 120, 578, 407 and 115 differentially expressed genes in liver, kidney, spleen, lung, trachea, hypothalamus, jejunum and heart, respectively. Interestingly, the changes in solute vector (SLC) family genes were found in all 8 tissues, and the verified gene results (SLC11A1, SLC17A7, SLC17A6, SLC6A4, SLC22A7, SLC25A3, SLC28A3, SLC7A2, SLC6A6, SLC38A5, SLC22A12, SLC34A1, SLC26A1, SLC26A6, SLC27A5, SLC22A8 and SLC44A4) were consistent with qRT-PCR results. In conclusion, ammonia exposure can cause pathological changes in many tissues and organs of fattening pigs and changes in the SCL family gene network. Importantly, the SCL family is involved in the toxic mechanism of ammonia. Our findings will provide a new insight for better assessing the mechanism of ammonia toxicity.

Protective effect of dietary supplementation of Bupleurum falcatum L saikosaponins on ammonia exposure-induced ileum injury in broilers.

Ammonia (NH3) at a high concentration has been recognized as a highly poisonous pollutant affecting both air and water quality. NH3, as a stimulus, exerts negative impact on broiler growth and production, but the molecular mechanisms are not clear yet. This study was designed to evaluate the effects of dietary supplementation of Bupleurum falcatum L saikosaponins (SP) on the growth and ileum health status in broilers exposed to NH3. Day-old Arbor Acers broilers (n = 480) were randomly allocated into 1 of 4 treatments. The main factors were dietary SP supplementation (0 or 80 mg/kg of diet) and NH3 challenge (with or without 70 ± 5 ppm NH3). The data of growth, intestinal morphology, and mRNA expression related to ileal function were collected from broilers exposed to NH3 for 7 d. Results showed that NH3 remarkably suppressed growth performance and intestinal development as well as induced biological injuries in the ileum of broilers, resulting from oxidative stress, mucous barrier damage, and immune dysfunction as well as upregulated apoptosis. These negative effects of NH3 were alleviated by the SP supplement. In conclusion, dietary supplementation of SP may be helpful in alleviating the detrimental effects of NH3 on the ileum development in broilers.

The protective effects of selenium on chronic ammonia toxicity in yellow catfish (Pelteobagrus fulvidraco).

Ammonia is toxic to most fish, and its negative effects can be eliminated by nutritional manipulation. In this study, triplicate groups of yellow catfish (0.58 ± 0.03 g) were fed diets supplemented with 0, 0.30 and 0.60 mg selenium (Se) kg-1 diet for 56 days under three ammonia contents (0.00, 5.70 and 11.40 mg L-1 total ammonia nitrogen). The results showed that ammonia toxicity could affects growth (weight gain, feed efficiency ratio, Se contents in muscle and whole body declined) and survival, leads to oxidative stress (total antioxidant capacity, superoxide dismutase, catalase and glutathione peroxidase activities declined and malondialdehyde accumulation), immunosuppression (lysozyme activity, 50% hemolytic complement, immunoglobulin M, respiratory burst and phagocytic index declined) and cytokines release (TNF, IL 1 and IL 8 elevated), induces up-regulation of antioxidant enzymes (Cu/Zn-SOD, Mn-SOD, CAT and GPx), cytokines (TNFα, IL 1 and IL 8) and pro-apoptotic genes (p53, Bax, Cytochrome c, Caspase 3 and Caspase 9) transcription, and down-regulation of anti-apoptotic gene Bcl2 transcription. The dietary Se supplementation could mitigate the adverse effect of ammonia poisoning on fish growth, oxidative damage, immunosuppression and apoptotic.

Improvement of growth, intestinal short-chain fatty acids, non-specific immunity and ammonia resistance in Pacific white shrimp (Litopenaeus vannamei) fed dietary water-soluble chitosan and mixed probiotics.

This study was to explore the impacts of water-soluble chitosan and mixed probiotics on growth performance, intestinal short-chain fatty acids (SCFAs) and immunity and ammonia resistance in Litopenaeus vannamei. Shrimp were fed one of four experimental diets including basal diet (CON), 0.10% water-soluble chitosan diet (WSC), 0.30% mixed probiotics (MP) and 0.10% water-soluble chitosan +0.30% mixed probiotics (SYN) for 8 weeks. Results showed shrimp fed with dietary MP and SYN diets could significantly improve growth performance and feed utilization in comparison with those of shrimp fed with CON diet (P < 0.05). Acetic acid content was significantly higher in shrimp fed with all supplemented diets compared to that in shrimp fed with CON diet (P < 0.05). Compared to shrimp fed with CON diet, dietary WSC and MP significantly influenced the contents and/or activities of aspartate aminotransferase (AST), total protein (TP), superoxide dismutase (SOD), lysozyme (LZM) in serum, SOD, malondialdehyde (MDA), acid phosphatase (ACP) in hepatopancreas and SOD and MDA in intestine. In addition, the gene expression levels of prophenoloxidase (proPO), penaiedin 3a (Pen-3a), crustin (Crustin), serine proteinase (SP), GPX and SOD in hepatopancreas, were significantly upregulated compared to those in CON diet at some time points (P < 0.05). Significantly higher survival rate in all supplemented diets were observed after ammonia challenge (P < 0.05). Therefore, the above results indicated dietary WSC and MP or SYN could enhance intestinal SCFAs content, stimulated antioxidant capacity and immune response, and increase the ammonia resistance of Litopenaeus vannamei. Besides, the growth performance was also improved by dietary MP and SYN.

ß-glucan extracted from eukaryotic single-celled microorganism Saccharomyces cerevisiae: Dietary supplementation and enhanced ammonia stress tolerance on Oreochromis mossambicus.

Ammonia is a widespread pollutant that is toxic to living organisms in aquaculture. This study aimed to evaluate the effects of a diet supplemented with beta-glucan from yeast, Saccharomyces cerevisiae (Sc-ßG), on the stress response of Oreochromis mossambicus (Tilapia) to ammonia. Fish were divided into four groups, including a control fed a basal diet and three experimental groups fed diets supplemented with Sc-ßG at 2, 5 and 10 mg/g respectively. After 8 weeks, experimental groups were exposed to ammonia at 100 mg L-1 for 1 week. Growth was measured after the 8-week feeding trial and serum, mucus, and liver tissue were sampled before and after the ammonia challenge. Compared with the control diet, feed supplemented with Sc-ßG at 10 mg/g significantly (p < 0.05) improved growth performance (7.8-9.9 g increase in weight). The cellular immune responses (myeloperoxidase, reactive oxygen species, and reactive nitrogen species), humoral immune responses (alkaline phosphatase, lysozyme, and peroxidase inhibition), and antioxidant response (catalase, superoxide dismutase, and glutathione) were tested in serum, mucus and liver tissue. Compared with the control, these responses were significantly (p < 0.05) enhanced at 10 mg/g supplementation with Sc-ßG. This study demonstrates that Sc-ßG may be applied to induce stress tolerance and improve growth performance in aquaculture.