Growth performance, mucosal immunity and disease resistance in goldfish (Carassius auratus) orally administered with Escherichia coli Strain Nissle 1917.

The current research aimed to shed light on the efficacy of Escherichia coli strain Nissle 1917 (EcN) on goldfish (……) growth, gut immunity, morphology, bacterial nutritional enzyme activity and resistance to Aeromonas hydrophila infection. Fish fed with EcN at 106, 107 and 108 CFU/g feed for 80 days showed an enhancement in growth better than control fish. The gut innate immunity in terms of lysozyme activity, immunoglobulin and total protein levels was increased in the treatment fish with the best result being observed in fish fed EcN at 108 CFU/ g. In addition, an increase was noted in the upregulation of immune-relevant genes, namely lysozyme, interleukin-1ß, inducible nitric oxide synthase and tumor necrosis factor α of fish intestine. A marked surge in the number of proteolytic and heterotrophic bacteria was noted in the gut of fish nourished with the probiotic. Histological studies exhibited an improvement in the intestinal absorption surface area, intraepithelial lymphocyte count and goblet cell density. Significantly higher survival rate was obtained in fish fed EcN at 108 CFU/g compared with the fish fed with the basal diet. These data exhibited the beneficial effect of EcN on goldfish growth, digestive enzymes, intestine heterotrophic bacteria and resistance against Aeromonas hydrophila challenge. This study confirmed the favorable outcomes resulting from the administration of EcN at108 CFU/g.

Immuno-modulatory Effects of Inactivated Dietzia Maris on the Selected Aspects of Cellular and Humoral Immune Responses in Mice.

Background: The current study is an attempt to register the alterations in the immunological and histological parameters in mice arising from the administration of Dietza maris (D. maris) in order to confirm its protective properties. Materials and Methods: Mice underwent 7 days of treatment with three doses of D. maris. Then, animals were scrutinized in terms of body weight, relative weight of organs, delayed type of hypersensitivity (DTH) response, and hemagglutination titer (HT). The determination of villus height, villus width, crypt depth, villus/crypt ratio (V/C), Goblet cells, and intestinal epithelial lymphocyte (IEL) density in villi was carried out. Results: A boosted DTH response was observed as a result of bacteria at medium dose. A variation was noted between the hemagglutinin titer of the control group and that of the high-dose group. Crypt depth, villus width, and villus height manifested alterations. High-dose-treated mice demonstrated proliferation of Goblet cells in the villi, whereas both in medium- and high-dose-treated mice, a distribution of IELs in the villus epithelium was noted. Overall, D. maris showed a stimulatory effect on immune functions in mice. Thus, thanks to improved cellular and humoral immunity and the increased quality of intestine function, we believe that D. maris promises novel therapeutic applications in the future. Conclusion: The attained findings lend credence to immuno-stimulatory effects arising from the capacity of D. maris to function as immunological adjuvants and to enhance humoral and cellular immunity as well as the intestinal structure and function.

Effects of Escherichia coli strain Nissle 1917 on arsenic-challenged goldfish (Carassius auratus): histological evidence.

Arsenic (As) contamination in natural water resources has become a great disaster throughout the world posing serious health problems. The current study was performed to evaluate the protective effects of Escherichia coli strain Nissle 1917 (EcN) against As exposure in goldfish (Carassius auratus). Fish were fed three times a day with 4.00% of body weight of diet with different doses (0.00, 1.00 × 106, 1.00 × 107 and 1.00 × 108 CFU g-1) of EcN for 80 days and then, challenged with 20.00 mg L-1 As for 96 hr under stagnant flow. Physicochemical characteristics of the inlet water were temperature of 25.10 ± 0.70 ˚C, pH of 7.30 ± 0.20 and dissolved oxygen of 7.30 ± 0.30 mg L-1 and 50.00% of water was exchanged once a week. Afterwards, fish were euthanized with a clove oil solution (50.00 µL L-1) and tissues were dissected from each fish and immediately fixed in 10.00% buffered formalin. The histopathological results indicated that the supplemented EcN did not have any side effects on various organs. It was also observed that the damages to kidney, liver, gill and skin were pronounced in fish exposed to As. However, the histopathological damages induced by As in fish tissues were less pronounced in the EcN-treated groups compared to the fish fed with the basal diet. Lamellar blood congestion in gills and epidermal cells detachment from the skin surface as well as hepatocytes, enterocytes and tubular necrosis were reduced in treated groups. These findings indicate that EcN has the potential to ameliorate the As-induced organ toxicity.

Efficacy of dietary live or heat-killed Bacillus subtilis in goldfish (Carassius auratus) infected with Ichthyophthirius multifiliis.

BACKGROUND: The beneficial effects of Bacillus subtilis on growth, immune response, and disease resistance against various diseases in different fish species have been proved. However, there are no data concerning this probiotic effect on skin mucosal immunity in fish infected with Ichthyophthirius multifiliis (Ich). Ich has a high mortality rate in both edible and ornamental fish and consequently is concerned with heavy economic losses. OBJECTIVES: Thus, we assessed the efficacy of live and heat-killed B. subtilis on skin immunity and histopathology in goldfish (Carassius auratus) infected with Ich. METHODS: Goldfish (144 fish, 2.38 g average weight) were stocked in nine glass tanks each in three replicates. Fish were fed 109 CFU g-1 live or heat-killed B. subtilis for 80 days. RESULTS: Probiotic administration in both viable and non-viable forms could enhance the growth performance in goldfish. Probiotic therapy also reduced the density of the parasite and histopathological level on skin and gill tissues of the treated fish. Real-time polymerase chain reaction analysis showed a higher expression of lysozyme and tumour necrosis factor-α in the treated groups compared to the control group. CONCLUSIONS: These data demonstrated the beneficial effect of B. subtilis as probiotic and paraprobiotic on growth performance and disease resistance to Ich infestation in goldfish.

Suppression of the malignancy of mammary tumor in mice model by inactivated preparation of Mycobacterium obuense.

Breast cancer (BC) is a significant cause of global mortality in women. This study was aimed to evaluate the immune-activation of malignant BC via the administration of attenuated Mycobacterium obuense. For this purpose, an in vivo model was developed with BALB/c mice. Mice were injected with 2.00 × 106 4T1 cells with breast tumor cell line. Forty-two mice were equally divided into control as well as low dose (0.20 mg 100 µL-1) and high dose (0.50 mg 100 µL-1) groups of M. obuense to investigate gene expression in the antitumor effects of M. obuense. In one group, paclitaxel was administrated as a choice drug in BC treatment. Antitumor manners were characterized by cytotoxicity against tumor target cells, size of the tumor and the expression of some BC metastatic genes together with pathology. The MTT assay demonstrated that different concentrations of both low and a high dose of bacteria did present no cytotoxicity effect on 4T1 cells. According to our findings, M. obuense significantly repressed tumor growth. M. obuense downregulated the expression of collagen type I alpha 1 (COLIA1), cFos, alkaline phosphatase (ALP), claudin 3 (cldn3), and conversely, activated transcription factor 4 (ATF4) and Twist related protein-1 (Twist1). All these alternations induced a decrease in the migratory and invasive capabilities of BC. The result of pathology was indicative of tumor regression in the paclitaxel and HK- M. obuense -recipient group. Thus, it seems most likely that M. obuense might impinge upon cell growth and metastatic behavior of malignant cells exerting anti-tumor activity in BC.

The Beneficial Effects of Actinomycetales Immune Modulators in the Pancreas of Diabetic Rats.

Purpose: Type 1 diabetes mellitus (T1DM) has dramatically increased in recent years, especially in young people, and limits the life quality of the patients involved. Thus, many researchers are performing extensive studies to find alternative treatments for DM. Methods: Here, we evaluated the improvement effects of the heat-killed Actinomycetales species, including Gordonia bronchialis, and Tsukamurella inchonensis in streptozotocin (STZ)- diabetic rats by biochemical, immunological, and histopathological examinations. Results: The present findings exhibited a dramatic and progressive alteration in the serum levels of interleukin-6 (IL-6), IL-10 and tumor necrosis factor-α (TNF-α) in the diabetic group, which were related to the blood glucose and insulin levels, oxidative stress defense (evaluated by TAC and MDA activities), and the pancreas biochemical indicators (such as amylase and lipase). More importantly, the present results were consistent with the histopathological findings, which included cellular degeneration, vascular congestion, hemorrhage, focal necrosis associated with mononuclear cell infiltration. Interestingly, all of the diabetic changes in the blood serum and tissues improved remarkably in the treated groups by Actinomycetales species. Conclusion: Surprisingly, most of the current diabetic complications effectively attenuated after oral administration of both Actinomycetales species, particularly with a high dose of T. inchonensis. Thus, it is concluded that the heat-killed Actinomycetales species can prevent and improve the progression of T1DM and its various complications profoundly.

The effects of heat-killed Tsukamurella inchonensis on intestinal morphology and humoral immune responses of broiler chickens.

BACKGROUND AND OBJECTIVES: Tsukamurella species are Gram-positive rods that exist in a broad range of environments. In this study, the efficacy of heat-killed Tsukamurella inchonensis on growth performance, intestinal morphology, and humoral immune responses of broiler chicken was evaluated. MATERIALS AND METHODS: Ross broiler chicks in the cage were randomly allocated to five groups. Trail diets were prepared by adding 106 cells per bird of heat-killed T. inchonensis into the basal trading diet for group 1 continuously dosed for 24 h from day 1 to day 13, and for group 2, 24 h on days 1 to 5; 8; 9, 12 and 13. Group 3 was received 106 bacteria as a subcutaneous injection on days 1, 6, and 12. Groups 4 and 5 were not received T. inchonensis during the experiment period. RESULTS: Feed intake (FI) and feed conversion ratio (FCR) were not altered by different delivery methods of T. inchonensis supplementation. The pulsed dosed in feed tended to provide higher body weight gain (BWG) than the negative control groups. T. inchonensis treatments, never less of the ways of delivery, boosted (P<0.05) the antibody titers to Newcastle disease virus (NDV), and avian influenza (AI) (H9N2) virus, especially when broiler chickens treated with pulse dosed in the feed. The most significant intestinal development (p<0.05) was observed between groups 1 and 2. There were no significant differences in the thymus, liver, and bursa of Fabricius relative weight. Still, there were significant increases in the relative weight of spleen on day 14 in vaccinated chickens treated with T. inchonensis pulse dosed. CONCLUSION: It seems that the supplementation of T. inchonensis in the broiler diet can improve intestinal morphology and humoral immune response, which was represented by increased antibody response to NDV, and AI vaccines significantly, but it cannot affect FI and FCR.

Treatment of mixed dairy and dye wastewater in anode of microbial fuel cell with simultaneous electricity generation.

Microbial fuel cell (MFC) is a green technology that converts the stored chemical energy of organic matter to electricity; therefore, it can be used for wastewater purification and energy production simultaneously. In this study, three kinds of dairy products, including milk, cheese water, and yogurt water, were mixed with Acid orange 7 (AO7) as the model wastewater and used as the anolyte of an MFC. The capability of the system in energy production and dye removal was also investigated. The FESEM images were used to investigate the biofilms attachment to the anodes. Moreover, the polarization curves, electrochemical impedance spectroscopy, cyclic voltammetry (CV), voltage-time profiles, and coulombic efficiency were used to evaluate the electrochemical activity of the MFCs. Based on the CV results, the biofilm formation significantly improved the electrochemical activity of the electrodes. Maximum power density, voltage, and coulombic efficiency were obtained as 44.05 mW.m-2, 332.4 mV, and 1.76%, respectively, for cheese water + AO7 anolyte, but the milk + AO7 MFC produced a stable voltage for a long time and its performance was similar to the cheese water + AO7 anolyte. Maximum COD removal and decolorization efficiencies were obtained equal to 84.57 and 92.18% for yogurt water + AO7 and cheese water + AO7 anolytes, respectively.

Immune enhancing effects of Lactobacillus acidophilus on Newcastle disease vaccination in chickens.

INTRODUCTION AND PURPOSE: Despite the various vaccination programs for protection against New Castle disease, it remains an important threat to the poultry industry. The ability of the probiotic bacteria to improve the immune system in both animals and humans supports their use as immune adjuvants for vaccination. The aim of the present study was to examine the effects of Lactobacillus acidophilus in ND vaccination. MATERIALS AND METHODS: A total of 170 one day old chicks were divided in 5 groups. In groups A, B and C chicks were received L. acidophilus (5 × 109, 3 × 109 and 2 × 109) and also vaccinated with inactivated and attenuated ND vaccines. In group D, chicks only vaccinated without bacterial inoculation and group E was negative control with neither vaccine nor bacteria. Then IgG and HI NDV antibody titers were measured in all tested groups. RESULTS: IgG and HI NDV antibody levels were significantly higher in Lactobacillus treated groups especially in group A with 5 × 109 bacteria, than only vaccinated and negative control groups. Also antibody levels against NDV increased during the vaccination period especially in probiotic treated groups. CONCLUSION: In conclusion, L. acidophilus can use for improving immunogenicity of NDV vaccination programs.

miR-330 Regulates Colorectal Cancer Oncogenesis by Targeting BACH1.

Purpose: Based on WHO report, colorectal cancer (CRC) is the second cause of death among patients with cancer worldwide. Dysregulation of miRNAs expressions has been demonstrated in different human cancers, especially CRC. Studies have shown that miR-330 could act as both TS-miR and/or oncomiR in different types of cancers. BACH1 is also identified as a transcription factor, which is involved in ontogenesis. In this study, we evaluated the CRC suppression via silencing of BACH1 by small silencer molecule called miR-330. Methods: Firstly, we analyzed the BACH1, miR-330-3p and miR-330-5p expressions according to the colon adenocarcinoma (COAD) and rectal adenocarcinoma (READ) project established from a patient of the colon and rectal cancer patients in The Cancer Genome Atlas (TCGA) database. The targeting of BACH1 via miR-330 in human CRC cells was evaluated by Vejnar bioinformatics methods, and confirmed by qRT-PCR and western blot analysis. Proliferation was performed by MTT assay. The MMP9, CXCR4, and VEGFR proteins were measured by western blotting. Results: The analysis of BACH1, miR-330-3p, and miR-330-5p expressions according to the COAD and READ projects showed that BACH1 was overexpressed, but miR-330-3p and miR330-5p were reduced in CRC tumors compared to normal controls. The miR-330 induction prevented proliferation of CRC cell by targeting BACH1 mRNA, which represses MMP9, C-X-C chemokine receptor type 4 (CXCR4), and vascular endothelial growth factor receptor (VEGFR) proteins expressions. Conclusion: Our results suggested that BACH1 is a potential target for miR-330 in CRC cells. The miR-330 induction inhibits CRC cells proliferation by suppressing BACH1 expression in posttranscriptional level. It was suggested that targeting of BACH1 via miRNA such as miR-330 could be a valid strategy in the field of CRC targeted therapy via modulating the oncogenic signaling pathway.

Beneficial effects of killed Tsukamurella inchonensis on rainbow trout (Oncorhynchus mykiss) growth, intestinal histology, immunological, and biochemical parameters.

Present study was conducted to investigate the effects of heat-killed Tsukamurella inchonensis on growth performance, gastrointestinal structure, immune response, and biochemical parameters in rainbow trout. Fish (mean weight 25 g) were fed basal diet (control), diets containing 2.48 × 108 colony-forming units (low-dose group) or 1.24 × 109 colony-forming units (high-dose group) of heat-killed Tsukamurella inchonensis per 1 kg of feed for 90 days. Results showed that growth performance was significantly enhanced in both treatment groups compared to the control group. The intestinal villus length and pyloric cecal fold length were mainly enhanced in the high-dose group. On the other hand, higher goblet cell percentage was shown with administration of dead Tsukamurella inchonensis in both treatment groups. Immune parameters such as alternative complement activity, immunoglobulin level, and hemagglutination titer were significantly higher in treatment groups than in fish fed in the control diet. Meanwhile, feeding heat-killed Tsukamurella inchonensis especially at higher dose caused a decrease in the levels of total cholesterol, uric acid, and lipid peroxidation product whereas no significant changes were noted in serum-specific marker enzymes levels, namely alanine aminotransferase (ALT), alkaline phosphatase (ALP), and aspartate aminotransferase (AST) by feeding both treatment diets compared to the control group. This study suggests that heat-killed Tsukamurella inchonensis especially at 1.24 × 109 colony-forming units had more potential to enhance growth, immunological parameters, and intestinal structure in rainbow trout.

Influence of extremely low frequency electromagnetic fields on growth performance, innate immune response, biochemical parameters and disease resistance in rainbow trout, Oncorhynchus mykiss.

The effects of extremely low frequency electromagnetic fields on rainbow trout growth performance, innate immunity and biochemical parameters were studied. Rainbow trout (17-18 g) were exposed to electromagnetic fields (15 Hz) at 0.01, 0.1, 0.5, 5 and 50 µT, for 1 h daily over period of 60 days. Growth performance of fish improved in different treatment groups, especially at 0.1, 0.5, 5 and 50 µT. Immunological parameters, specifically hemagglutinating titer, total antiprotease and α1-antiprotease levels in treatment groups, were also enhanced. Total protein and globulin contents in the serum of fish exposed to 0.1, 0.5, 5 and 50 µT were significantly higher than those in the control group. No significant differences were found in serum enzyme activities, namely aspartate aminotransferase and alanine aminotransferase of fish in all treatment groups. Conversely, alkaline phosphatase level decreased in fish exposed to 0.01 and 50 µT electromagnetic fields. Meanwhile, electromagnetic induction at 0.1, 0.5, 5 and 50 µT enhanced fish protection against Yersinia ruckeri. These results indicated that these specific electromagnetic fields had possible effects on growth performance, nonspecific immunity and disease resistance of rainbow trout.

Green tea (Camellia sinensis) administration induces expression of immune relevant genes and biochemical parameters in rainbow trout (Oncorhynchus mykiss).

Present study elucidates the efficacy of green tea (Camellia sinensis) on growth performance, immune and antioxidant systems and cytokine gene expression in rainbow trout tissues. Green tea was supplemented at 20, 100, and 500 mg kg(-1) diet and fed to fish (average weight: 23.5 g) for 35 days. No remarkable changes in growth performance were observed among all test groups. Lower lipid peroxidation product and higher superoxide dismutase activity were noted in fish received the medium dose of green tea. Significant increase in serum bactericidal activity and total protein were recorded in all treatment groups. All doses of green tea up-regulated Interleukin-1ß transcription in the spleen, while Interleukin-1ß mRNA level decreased significantly in the kidney of low dose of green tea. Interleukin-6 mRNA level was up-regulated in the spleen of high dose of green tea and liver of middle and high doses of green tea. High dose and medium dose of green tea up-regulated the interleukin-8 transcription in the kidney and liver, respectively. Meanwhile, green tea inhibited the production of interleukin-10 in all treatment groups compared with control group. Medium dose of green tea up-regulated tumor necrosis factor-α transcription in all fish tissues, while high dose and low dose of green tea enhanced tumor necrosis factor-α mRNA levels in the kidney and spleen, respectively. Present study suggests that green tea especially at 100 mg kg(-1) feed may effectively enhance the antioxidant system and immune system in rainbow trout.

Hilyses®, fermented Saccharomyces cerevisiae, enhances the growth performance and skin non-specific immune parameters in rainbow trout (Oncorhynchus mykiss).

Effects of Hilyses(®), fermented Saccharomyces cerevisiae (S. cerevisiae), on growth, body composition and skin mucus immune components in rainbow trout were quantified. Ninety rainbow trout (105 ± 5 g) were randomly assigned to 2 groups in triplicates and fed dietary Hilyses(®) (5 g kg(-1)) or control diet without Hilyses(®) for 50 days. Results of this study demonstrated that growth performance increased significantly by the dietary yeast supplement; however body composition was not affected in treatment group. At the 45th and 50th day of feeding trial, results of mucus samples demonstrated that yeast supplementation in treatment group significantly promoted enzyme activities, namely lysozyme, protease, alkaline phosphatase and esterase compared to control group. Significant increases were also observed in hemagglutination and antibacterial activity against Yersinia ruckeri in fish fed treatment diet. The present study suggests that fermented S. cerevisiae may effectively promote the growth performance and skin non-specific immune parameters in rainbow trout.

Effects of dietary Ergosan on cutaneous mucosal immune response in rainbow trout (Oncorhynchus mykiss).

The effects of dietary Ergosan on the growth performance and mucosal immunity in rainbow trout skin were investigated. 60 rainbow trout (100-110 g) were randomly assigned to 2 groups in triplicates and fed one of the experimental diet formulated with 5 g kg⁻¹ Ergosan or control diet for 50 days. Results showed that on the 45th day of feeding trial, Ergosan supplementation significantly enhanced the growth performance compared to control group. Various enzyme activities, namely lysozyme, protease, alkaline phosphatase and esterase in treatment group were also enhanced on the 45th and 50th day. Skin mucus in Ergosan-fed fish showed the agglutination of erythrocytes while in control group, no visible agglutination was shown. In addition, skin mucus in treatment group showed strong antibacterial activity against Yersinia ruckeri. In conclusion, the major immune components of rainbow trout mucus that are involved in the non-specific immunity were enhanced by administration of Ergosan in 5 g kg⁻¹.

Immunomodulatory effects of decaffeinated green tea (Camellia sinensis) on the immune system of rainbow trout (Oncorhynchus mykiss).

In order to study the immunomodulatory effects of decaffeinated green tea extract on rainbow trout, a study with a 30-day feeding trial was conducted. Commercial diets with graded levels of decaffeinated green tea extract, 20 mg (T1), 100 mg (T2), 500 mg (T3) per kg feed were prepared. 120 rainbow trout (35 ± 3 g) were randomly assigned to 4 groups in triplicates and fed one of the 3 experimental diets formulated or control diet. After feeding trial, 12 fish from each group were sampled for analysis of some immunological parameters. Remaining fish were injected with 0.5 ml of chicken red blood cell (C-RBC) suspension (2%) intraperitoneally on days 5 and 15 after feeding trial. Results of the current study showed that the inclusion of 20 mg kg-1 green tea (T1) in fish diet enhanced the serum bactericidal activity against Yersinia ruckeri, while significant elevation of lysozyme activity was shown in T2 group. Anti-trypsin activity due to α1-antiprotease was significantly higher in T1 and T2 groups while peroxidase content showed significant increase in all treatment groups compared to control group. Hemagglutination antibody titer against C-RBC was significantly higher in fish administered with 100 mg kg(-1) green tea (T2). Our findings showed that decaffeinated green tea in lower doses of administration could be optimum to enhance the immunity of rainbow trout.