Dragon fruit-derived oligosaccharides alter hemocyte-mediated immunity and expression of genes related to innate immunity and oxidative stress in Daphnia magna.

Dragon fruit oligosaccharide (DFO) is an indigestible prebiotic that enhances the growth and reproduction of Daphnia magna, increases the expression of genes involved in immunity, and reduces oxidative stress. This study investigated the effects of DFO on the expression of innate immunity- (Toll, Pelle, proPO, A2M, and CTL), oxidative stress- (Mn-SOD), and nitric oxide (NO) synthesis-related genes (NOS1, NOS2, and arginase) as well as NO localization and number of hemocytes in D. magna. For this ten-day-old D. magna were treated with 0 or 9 mg l-1 of DFO for 24 and 85 h. Gene expression levels, NO intensity and localization, and total hemocytes were evaluated. After 24 h, the expression of Toll and proPO increased significantly (p < 0.05), while that of C-type lectins (CTL) was reduced (p < 0.05). At 85 h, Mn-SOD and CTL expressions were markedly suppressed (p < 0.05). NO was mostly localized in the foregut, midgut, hindgut, and carapace. The expression of NOS1 was reduced after 24 h (p < 0.05). In addition, NO intensity at 24 h was insignificantly lower than the control (p > 0.05). At 85 h, the expression of NOS1, NOS2, and arginase was higher than control, but NO intensity did not differ significantly (p > 0.05). Furthermore, the total hemocyte count elevated remarkably at 85 h (p < 0.05). Our study suggested that 9 mg l-1 of DFO could alter the expression of the genes related to innate immunity, oxidative stress, and NO synthesis in D. magna and significantly stimulate hemocyte production.

Combined effects of nanoplastics and elevated temperature in the freshwater water flea Daphnia magna.

Global warming and nanoplastics (NPs) are critical global issues. Among NPs, one of the most hazardous types of plastics, polystyrene (PS), poses ecotoxicological threats to several freshwater organisms. The degree of toxicity of PS-NPs is strongly influenced by various environmental factors. This study illustrates the combined effects of temperature and PS-NPs on the water flea Daphnia magna. The sensitivity of D. magna to PS-NPs was tested under control (23 °C) and elevated temperatures (28 °C). As a result, increased temperatures influenced the uptake and accumulation of PS-NPs. Co-exposure to both higher temperatures and PS-NPs resulted in a drastic decrease in reproductive performance. The level of oxidative stress was found to have increased in a temperature-dependent manner. Oxidative stress was stimulated by both stressors, leading to increased levels of reactive oxygen species and antioxidant enzyme activity supported by upregulation of antioxidant enzyme-related genes under combined PS-NPs exposure and elevated temperature. In the imbalanced status of intracellular redox, activation of the p38 mitogen-activated protein kinase signaling pathway was induced by exposure to PS-NPs at high temperatures, which supported the decline of the reproductive capacity of D. magna. Therefore, our results suggest that PS-NPs exposure along with an increase in temperature significantly affects physiological processes triggered by damage from oxidative stress, leading to severely inhibited reproduction of D. magna.

Thermal stress-stimulated ZnO toxicity inhibits reproduction of freshwater crustacean Daphnia magna.

Elevated temperatures due to climate change pose a variety of environmental risks to the freshwater ecosystem. At the same time, zinc oxide (ZnO) has become widely used and has entered the freshwater environment. As thermal stress may potentially impact the physicochemical properties of ZnO, its toxicity to freshwater organisms in the face of global warming is poorly understood. The potential effects on reproductive performances, including oogenesis, are of particular concern. In this study, we investigate the reproductive performances and related mRNA abundance of the zooplankton Daphnia magna under conditions of ZnO exposure and heat stress. The results revealed that ZnO and elevated temperature delayed maturity and juvenile production of D. magna. Histological observations indicated that oogenesis was inhibited, and the number and size of oocytes were reduced in the condition of ZnO exposure under heat stress. Eventual offspring in the same treatment exhibited decreased numbers, size, and quality. Congenital juvenile anomalies were increased, such as deformed eye, and impaired antenna and tail spine. Moreover, both ZnO and elevated temperature treatments inhibited expression levels of reproduction-related genes (vtg, EcR and VMO1) and induced the dmrt93b gene involved in the production of male offspring. Furthermore, we found that D. magna tried to cope with ZnO and thermal stress by upregulating hsp90, HIF-1α and HIF-1ß. ZnO and heat stress inhibited the reproductive capacity of D. magna, produced deleterious effects on reproduction-associated physiological pathways, and damaged reproductive outcomes.

Ecotoxicological, ecophysiological, and mechanistic studies on zinc oxide (ZnO) toxicity in freshwater environment.

The world has faced climate change that affects hydrology and thermal systems in the aquatic environment resulting in temperature changes, which directly affect the aquatic ecosystem. Elevated water temperature influences the physico-chemical properties of chemicals in freshwater ecosystems leading to disturbing living organisms. Owing to the industrial revolution, the mass production of zinc oxide (ZnO) has been led to contaminated environments, and therefore, the toxicological effects of ZnO become more concerning under climate change scenarios. A comprehensive understanding of its toxicity influenced by main factors driven by climate change is indispensable. This review summarized the detrimental effects of ZnO with a single ZnO exposure and combined it with key climate change-associated factors in many aspects (i.e., oxidative stress, energy reserves, behavior and life history traits). Moreover, this review tried to point out ZnO kinetic behavior and corresponding mechanisms which pose a problem of observed detrimental effects correlated with the alteration of elevated temperature.

Probiotics of Lacticaseibacillus paracasei SD1 and Lacticaseibacillus rhamnosus SD11 attenuate inflammation and ß-cell death in streptozotocin-induced type 1 diabetic mice.

Probiotics provide health benefits in various aspects and are believed to modulate the immune system by balancing gut microbiota homeostasis, termed the "microbiota-immune axis". Recent evidence supports that several Lactobacillus strains possess glucose-lowering and anti-inflammatory effects in an animal model of type 1 diabetes (T1D). Although probiotics of Lacticaseibacillus paracasei SD1 (SD1) and Lacticaseibacillus rhamnosus SD11 (SD11) exert human oral health benefits by reducing harmful bacterial populations, their clinical application regarding hypoglycemic-related traits as well as the underlying mechanisms are still lacking. In this report, we used multiple low doses of streptozotocin (STZ)-induced diabetic BALB/c mice to explore the effects of SD1 and SD11 supplementation on the regulation of markers related to T1D. Experimental mice were randomly assigned into five groups, non-STZ + V, STZ + V, STZ + SD1, STZ + SD11, and STZ + SDM (mixture of SD1 and SD11), and physiological data were measured every week. Blood and pancreas samples were collected at 4- and 8-weeks. Our results indicate that supplementation with SD1, SD11, or SDM for 8 weeks significantly improved body weights, glycemic levels, glucose tolerance, insulin levels, and lipid profiles. Probiotic administration also preserved islet integrity and increased ß-cell mass in STZ-injected mice, as well as prevented infiltration of macrophages, CD4+, and CD8+ T cells into the islets. Significantly, SD1 and SD11 suppressed the levels of IL1-ß, TNF-α and IFN-γ and increased IL-10, which is concomitant with the inhibition of cleaved caspase 3, caspase 9, caspase 8, proapoptotic Bax, NF-κBp65, pSTAT1, and iNOS. Additionally, the survival ability of ß-cells was mediated by upregulated anti-apoptotic Bcl2. We conclude that SD1 and SD11 attenuate STZ-induced diabetic mice by stabilizing glycemic levels and reducing inflammation, thereby protecting ß-cells. Among the probiotic treatment groups, SD11 revealed the best results in almost all parameters, indicating its potential use for alleviating hyperglycemia-associated symptoms.

Disturbances in growth, oxidative stress, energy reserves and the expressions of related genes in Daphnia magna after exposure to ZnO under thermal stress.

The toxicological effects of metal contamination are influenced by the ambient temperature. Therefore, global warming affects the toxicity of metal contamination in aquatic ecosystems. ZnO is widely used as a catalyst in many industries, and causes contamination in aquatic ecosystems. Here, we investigated the effects of ZnO concentration under elevated temperature by observing growth, oxidative stress, energy reserves and related gene expression in exposed Daphnia magna. Body length and growth rate increased in neonates exposed to ZnO for 2 days but decreased at 9 and 21 days under elevated temperature. ZnO concentration and elevated temperature induced oxidative stress in mature D. magna by reducing superoxide dismutase (SOD) activity and increasing malondialdehyde (MDA) levels. In contrast, juveniles were unaffected. Carbohydrate, protein and caloric contents were reduced throughout development in D. magna treated with ZnO and elevated temperature in all exposure periods (2, 9 and 21 days). However, lipid content also decreased in mature D. magna treated with ZnO cultured under elevated temperature, while that of juveniles showed an increase in lipid content. Therefore, energy was perhaps allocated to physiological processes for detoxification and homeostasis. Moreover, expression patterns of genes related to physiological processes changed under elevated temperature and ZnO exposure. Taken together, our results highlight that the combination of temperature and ZnO concentration induced toxicity in D. magna. This conclusion was confirmed by the Integrated Biological Response (IBR) index. This study shows that changes in biological levels of organization could be used to monitor environmental change using D. magna as a bioindicator.

Sargassum plagiophyllum Extract Enhances Colonic Functions and Modulates Gut Microbiota in Constipated Mice.

Constipation is a symptom that is widely found in the world's population. Various dietary supplementations are used to relieve and prevent constipation. Seaweed is widely used for its health benefits. In this study, we aimed to investigate the effects of Sargassum plagiophyllum extract (SPE) on functions of the gastrointestinal tract and gut microbiota. The results show that SPE pretreatment increased the frequency of gut contraction, leading to reduce gut transit time. SPE pretreatment also significantly increased the secretion of Cl− and reduced Na+ absorption, increasing fecal water content in constipated mice (p < 0.05). In addition, the Bifidobacteria population in cecal contents was significantly higher in constipated mice pretreated with 500 mg/kg SPE for 14 days than in untreated constipated mice (p < 0.05). Our findings suggest that SPE can prevent constipation in loperamide-induced mice. This study may be useful for the development of human food supplements from S. plagiophyllum, which prevent constipation.

Bacteria Associated with Echinodorus cordifolius and Lepironia articulata Enhance Nitrogen and Phosphorus Removal from Wastewater.

Phytoremediation and bioremediation are eco-friendly methods of wastewater treatment that are widely used throughout the world to reduce anthropogenic water contamination. This study was conducted to assess the effectiveness of symbiotic bacteria in phytoremediation using two aquatic plants, Echinodorus cordifolius and Lepironia articulata, that were tested in sterilized and unsterilized groups. The results showed that unsterilized plants removed more phosphate, ammonium, nitrate and nitrite than the sterilized plants. In untreated and unsterilized E. cordifolius groups, the dominant bacterium was Calothrix (46.90 and 49.69%, respectively), which was higher than in the sterilized E. cordifolius group (38.88%). In untreated and unsterilized groups of L. articulata, Clostridium was a dominant bacterium. The proportion of Clostridium was much lower in the sterilized L. articulata group (1.31%) than in the untreated (13.71%) and unsterilized (49.02%) groups. Our results suggested that root-associated bacteria in E. cordifolius and L. articulata were effective in the removal of phosphorus and nitrogen from domestic wastewater.

Temperature and concentration of ZnO particles affect life history traits and oxidative stress in Daphnia magna.

Temperature affects physiological processes in organisms and the toxicity of chemicals. The widespread industrial use of ZnO causes contamination in aquatic ecosystems. This study aimed to investigate the chronic toxicity of ZnO at different temperatures using Daphnia magna as a model organism. The chronic toxicity of five different concentrations of ZnO was assessed at 23 °C and 28 °C. The results showed that higher concentrations of ZnO inhibited growth, production of first clutch eggs and juvenile accumulation at both 23 °C and 28 °C. Growth rate, numbers of first clutch eggs and juvenile accumulation were lower at 28 °C than at 23 °C. We also observed the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) activity. At higher concentrations of ZnO, oxidative stress was induced leading to increase MDA level and decrease SOD activity at 28 °C. These findings indicated that high temperature and high concentration of ZnO inhibited the activity of enzymatic proteins. Nonetheless, among all treatments, the accumulation of zinc in D. magna was not significantly different. Our results suggested that both ZnO and higher temperature induced oxidative stress in D. magna. As a result, MDA concentration increased, SOD activity changed and the growth and reproduction of D. magna was adversely affected.

Oligosaccharides derived from dragon fruit modulate gut microbiota, reduce oxidative stress and stimulate toll-pathway related gene expression in freshwater crustacean Daphnia magna.

Dragon fruit oligosaccharide (DFO) is an indigestible prebiotic. In this study, we aimed to investigate the effects of DFO on gut microbiota, oxidative stress and immune-related gene expression in Daphnia magna. The 10-day-old D. magna were treated with 0, 9, and 27 mg l-1 DFO for 85 h. The gut bacterial communities, superoxide dismutase (SOD) activity, lipid peroxidation and the expressions of genes in Toll signaling pathway were observed. The results showed that D. magna treated with 9 and 27 mg l-1 DFO altered gut microbiota composition by increasing Limnohabitans and Lactobacillus, and significantly increased SOD activity and reduced lipid peroxidation. Moreover, the expressions of Toll2, Toll3, Toll5, Toll7 and Pelle genes were significantly increased in D. magna treated with 9 and 27 mg l-1 DFO. Our results suggested that DFO changed the composition of the gut microbiota of D. magna by increasing the beneficial bacteria. DFO also had the ability to stimulate innate immunity in D. magna by increasing SOD activity, reducing lipid peroxidation, and increasing the expression of immune-related genes.

Prebiotic oligosaccharides from dragon fruits alter gut motility in mice.

Dragon fruit oligosaccharide (DFO) has a prebiotic property which improves gut health by selectively stimulating the colonic microbiota. Altering microbiota composition may affect intestinal motility. However, no study has been done to understand the DFO effects on gut motor functions. This research thus aimed to investigate the DFO effects on mice colons compared to the prebiotic fructo-oligosaccharide (FOS) and probiotic bifidobacteria. The mice in this study received distilled water; 100, 500, and 1000 mg/kg DFO; 1000 mg/kg FOS; or 109 CFU Bifidobacterium animalis daily for 1 week and some treatments for 2 weeks. Gastrointestinal transits were analysed and motility patterns, smooth muscle (SM) contractions and morphological structures of the colons were assessed. Administration of FOS, 500 and 1000 mg/kg DFO significantly increased fecal output when compared to the control group. In mice treated with FOS and bifidobacteria, gut transit time was reduced, while upper gut transit was increased in comparison to DFO groups. Spatiotemporal maps of colonic wall motions showed that DFO increased the number of colonic non-propagation contractions and fecal pellet velocity, consistent with the results from groups treated with FOS and bifidobacteria. DFO also increased the amplitude and duration of colonic SM contractions. Histological stains showed normal epithelia, crypts, goblet cells, and SM thickness in all groups. In conclusion, DFO increased colonic SM contractions without morphological change and acted as a bulk-forming and stimulant laxative to increase fecal output and intestinal motility. Thus, DFO as a dietary supplement may promote gut health and correct gastrointestinal motility disorders.

Effects of symbiotic bacteria on chemical sensitivity of Daphnia magna.

The crustacean zooplankton Daphnia magna has been widely used for chemical toxicity tests. Although abiotic factors have been well documented in ecotoxicological test protocols, biotic factors that may affect the sensitivity to chemical compounds remain limited. Recently, we identified symbiotic bacteria that are critical for the growth and reproduction of D. magna. The presence of symbiotic bacteria on Daphnia raised the question as to whether these bacteria have a positive or negative effect on toxicity tests. In order to evaluate the effects of symbiotic bacteria on toxicity tests, bacteria-free Daphnia were prepared, and their chemical sensitivities were compared with that of Daphnia with symbiotic bacteria based on an acute immobilization test. The Daphnia with symbiotic bacteria showed higher chemical resistance to nonylphenol, fenoxycarb, and pentachlorophenol than bacteria-free Daphnia. These results suggested potential roles of symbiotic bacteria in the chemical resistance of its host Daphnia.

Betaproteobacteria Limnohabitans strains increase fecundity in the crustacean Daphnia magna: symbiotic relationship between major bacterioplankton and zooplankton in freshwater ecosystem.

How symbioses between bacteria and aquatic animals influence food webs in freshwater ecosystems is a fundamental question in ecology. We investigated symbiosis between a crustacean zooplankton Daphnia magna and its dominant bacterial symbiont Limnohabitans, an abundant and globally distributed freshwater Betaproteobacteria. Aposymbiotic juvenile Daphnia were prepared and exposed to any of four Limnohabitans sp. - Limnohabitans strains DM1, 2KL-3, 2KL-7 and Limnohabitans planktonicus strain II-D5, all previously found in D. magna digestive tract or culture. Re-infected Daphnia were cultured until they produced the first clutch of juveniles. Limnohabitans strain DM1 and L. planktonicus strain II-D5 successfully re-infected Daphnia through single exposure at the first instar juvenile stage. In contrast to aposymbiotic Daphnia that produced non-viable juveniles, re-infected Daphnia produced viable juveniles and increased fecundity to levels of that of symbiotic Daphnia. Re-infected Daphnia did not increase their number of eggs nor growth rates. Limnohabitans strains 2KL-7 and 2KL-3 could not recover fecundity even in multiple exposures during culture. This study shows the functional evidence demonstrating that a single bacterium Limnohabitans regulates fecundity of the consumer Daphnia through symbiosis. Our results indicated that symbiotic relationship between major bacterioplankton and zooplankton is important for maintaining the population of zooplankton in freshwater ecosystems.

Symbiotic bacteria contribute to increasing the population size of a freshwater crustacean, Daphnia magna.

The filter-feeding crustacean Daphnia is a key organism in freshwater ecosystems. Here, we report the effect of symbiotic bacteria on ecologically important life history traits, such as population dynamics and longevity, in Daphnia magna. By disinfection of the daphniid embryos with glutaraldehyde, aposymbiotic daphniids were prepared and cultured under bacteria-free conditions. Removal of bacteria from the daphniids was monitored by quantitative polymerase chain reaction for bacterial 16S rRNA gene. The population of aposymbiotic daphniids was reduced 10-folds compared with that of the control daphniids. Importantly, re-infection with symbiotic bacteria caused daphniids to regain bacteria and increase their fecundity to the level of the control daphniids, suggesting that symbiotic bacteria regulate Daphnia fecundity. To identify the species of symbiotic bacteria, 16S rRNA genes of bacteria in daphniids were sequenced. This revealed that 50% of sequences belonged to the Limnohabitans sp. of the Betaproteobacteria class and that the diversity of bacterial taxa was relatively low. These results suggested that symbiotic bacteria have a beneficial effect on D. magna, and that aposymbiotic Daphnia are useful tools in understanding the role of symbiotic bacteria in the environmental responses and evolution of their hosts.