Prospective use of bacteriocinogenic Pediococcus pentosaceus as direct-fed microbial having methane reducing potential

Direct-fed microbials(DFM), generally regarded as safe status, are successfully used in improving rumen ecology, gastro-intestinal health, feed efficiency, milk production and growth rate in ruminants. On the other hand, methanogenesis in rumen, which accounts for a significant loss of ruminant energy and increased greenhouse gas in environment, is of great concern, therefore, use of DFM for improving productivity without compromising the animal health and ecological sustainability is encouraged. The present study was conducted to investigate the methane reducing potential of bacteriocinogenic strain Pediococcus pentosaceus-34. Since, the culture showed no hemolysis on blood agar and DNase activity, hence, it was considered to be avirulent in nature, a prerequisite for any DFM. The culture also showed tolerance to pH 5.0 for 24 h with 0.5% organic acid mixture, whereas when given a shock for 2 h at different p H and organic acids concentrations, it showed growth at pH 3.0 and 4.0 with 0.1 and 1.0% organic acids, respectively, as having good animal probiotics attributes. The total gas production was significantly(P<0.05) higher in live pedicoccal culture(LPC) and dead pedicoccal culture(DPC) both with wheat straw, when compared to the control. In sugarcane bagasse, gas production was significantly lower(P<0.05) with LPC compared to the control and DPC both. Methane was reduced by the inclusion of LPC in sugarcane bagasse(0.07 mL CH4 mg–1 dry matter digestibility) with no effect on other rumen fermentation parameters. However, with wheat straw and LPC total gas, in vitro dry matter digestibility, total volatile fatty acids increased significantly but no reduction in methane production was observed in comparison to the control. Therefore, further research is warranted in this direction, if the bacteriocinogenic strains can be used as DFM for ruminants to improve the ruminant productivity.     

Chrysin Reduced Acrylamide-Induced Neurotoxicity in Both in vitro and in vivo Assessments

Background: Acrylamide (ACR) is a well-known industrial toxic chemical that produces neurotoxicity, which is characterized by progressive central and peripheral neuronal degeneration. Chrysin is a natural, biologically active flavonoid compound, which is commonly found in many plants. The antioxidant and neuroprotective properties of chrysin have been demonstrated. Methods: In this study, the possible effect of chrysin on ACR-induced toxicity was evaluated in both in vitro and in vivo experiments. PC12 cells were used as a suitable in vitro model. Cells were exposed to chrysin (0.5-5 µM) for 12 and 24 h, and then ACR in IC₅₀ concentration was added to the cells. Finally, cell viability was determined using (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium assay. For in vivo assay, Wistar rats were treated with ACR (50 mg/kg i.p. for 11 days) alone or in combination with chrysin (12.5, 25, and 50 mg/kg). At the end of treatment, behavioral index was evaluated. Results: ACR decreased cell viability and pre-treatment with chrysin (0.5-5 µM) significantly decreased ACR-induced cytotoxicity in the time- and dose-dependent manner. In Wistar rats, exposure to ACR significantly induced severe gait abnormalities, but treatment with chrysin (50 mg/kg) reduced ACR-induced neurotoxicity in animals. Conclusion: In the current study, chrysin exhibited neuroprotective effect on PC12 cells as an in vitro model and also on Wistar rats. Iran. Key Words: Acrylamide, Chrysin, Neurotoxicity, Antioxidant     

Effect of body mass index on severity and prevalence of varicocele

Varicocele is classified as grade I-III regarding its severity. This study was aimed to determine the correlation between height and weight with varicocele grade in 18-30 years age group. We enrolled 400 persons aged 18-30 years referred to the specialist's clinics of Tabriz Medical Sciences University or Medical Commission Since Sep. 2004 to Mar. 2005. First we divided the volunteers in two groups including Varicocele Group and Non-varicocele Group, then varicocele patients were classified to three grades considering the severity of the disease: severe (Grade III), moderate (Grade II) and mild (Grade I). Finally, the correlation between height, weight and Body Mass Index (BMI) was evaluated. There was a significant relation between height and grades of left-side varicocele; in other words the severity of disease was increased with height (p = 0.004). Also, height increased the prevalence ofvaricocele (p = 0.011). On the other hand, low weight and BMI increased the prevalence of varicocele (p = 0.000, p = 0.004) but did not affect the severity of disease (p = 0.364, p = 0.172). In conclusion, the height of patients directly affected the prevalence and severity of left-side varicocele which probably is related to length of left internal spermatic veins in these patients and increased hydrostatic pressure in taller patients. Also, the weight and BMI is effective on the prevalence of varicocele. It seems that slim and tall persons will benefit from evaluation while puberty.     

Yield Components and Fatty Acids Variation of Canola Cultivars Under Different Irrigation Regimes and Planting Dates

To investigate the effect of different planting dates and irrigation regimes on six canola cultivars, a 2-year (2014–2016) experiment was conducted at the Seed and Plant Improvement Institute of Karaj. The experiments were conducted as the factorial split-plot in a randomized complete block design including six canola cultivars (Gabriella, Brutus, Triangle, Marathon, Danube, and Natali), two irrigation regimes (fully irrigated and irrigation termination at the flowering stage) and two planting dates (October 1 and November 1). The results showed that irrigation termination from the flowering stage prevented the supply of required material for filling the seeds and the metabolism of the seed compounds and reduced the growth period of the seed. Therefore, a reduction was observed in the content of oleic and linoleic acids and proline. Also, water deficit stress caused an increase in the glucosinolate content of the seed. The response of canola cultivars was different in terms of fatty acids, so that the Natali cultivar had higher palmitic, oleic acids, and proline content compared to the other cultivars at both planting dates. Fully irrigated treatment and planting date of October 1 produced the highest amount of seed and oil yield (4659 and 2073?kg ha^?1), palmitic acid (5.5%), and proline (22.7?μmol/g) content, and the lowest glucosinolate content (13.2?mg/g). Generally, the Natali cultivar and planting date of October 1 and the fully irrigated regime are highly recommended in the studied area regarding qualitative traits and yield of canola.

     

Molecular and morphological investigation in Hymenocrater: species delimitation,relationship, divergence time and DNA barcoding

Genetic Resources and Crop Evolution - Hymenocrater comprises of 11 species throught the world. The genus is an Irano-Turanian elements, which has several usages in traditional medicine. Northeast...     

Inhibitory effects of Sage extract on the growth of enteric bacteria

Antibacterial activity of Sage extract at concentrations of 0.1, 0.05, 0.025, 0.0125, 0.00625, 0.003125, 0.00156, 0.0005 and 0.00025 g dL(-1) against Salmonella typhi, Shigella sonnei, S. flexneri, Proteus vulgaris, Staphylococcus aureus, ETEC Escherichia coli and Pseudomonas aeruginosa was evaluated. Susceptibility testing of bacterial strains against 18 antibiotics was also performed for comparison. The results showed that P. aeruginosa and ETEC E. coli were completely resistant to Sage extract even at concentration of 0.1 g dL(-1). Its antibacterial activity (0.1 g dL(-1)) against P. vulgaris, S. flexneri and S. sonnei was the same as nitrofurantoin and ampicilline respectively. Sage extract (0.1 and 0.05 g dL(-1)) exhibited the same effects as ampicilline and streptomycin against S. typhi. Its antibacterial activity (0.1, 0.05 and 0.25 g dL(-1)) against S. aureus was the same as ceftazidim, chloramphenicol, gentamycin, neomycin and nitrofurantoin and was more significant compared to streptomycin and vancomycin. The results suggest Sage can be considered as an alternative herbal in the treatment of infections caused by the above-mentioned bacteria.     

Inotropic and chronotropic effects of 6-hydroxy-4-methylquinolin-2(1H)-one derivatives in isolated rat atria

Background: Selective phosphodiesterase (PDE3) inhibitors improve cardiac contractility and may use in congestive heart failure. However, their proarrhythmic potential is the most important side effect. Methods: In this research, we evaluated the potential cardiotonic activity of six new synthesized selective PDE3 inhibitors (6-hydroxy-4-methylquinolin-2(1H)-one derivatives) using the spontaneously beating atria model. In each experiment, atrium of reserpine-treated rat was isolated and the contractile and chronotropic effects of a synthesized compound were assessed. The 3-isobutyl-1-methylxanthine, a non-selective PDE inhibitor, was used for comparison. Results: The results showed that, among new compounds, the best pharmacological profile was obtained with the compound 6-[4-(4-methylpiperazine-1-yl)-4-oxobutoxy]-4-methylquinolin-2(1H)-one, C6, which displayed selectivity for increasing the force of contraction (168 +/- 5% change over the control) rather than the frequency rate (138 +/- 5% change over the control) at 300 muM. However, C6 at concentrations of 10 and 100 muM produced left and upward shift in the positive inotropic concentration-response curve of isoprenaline. The -log EC50 of isoprenaline was 8.843 +/- 0.171 in the absence, 9.448 +/- 0.138 and 9.456 +/- 0.107 in the presence of 10, 100 muM of C6, respectively (P<0.001, n = 6). Also, amrinone, a selective PDE3 inhibitor, shifted the isoprenaline concentration-response curve to the left and upward. The concentration of 10 and 100 muM amrinone decreased -log EC50 of isoprenaline to 9.527 +/- 0.287 and 9.423 +/- 0.243, respectively (P<0.001, n = 6). Moreover, the positive chronotropic effect of isoprenaline was not affected by amrinone or C6. Conclusion: This study provides functional evidence for the positive inotropic effect of C6. Considering the augmentation of isoprenaline positive inotropic concentration-response with C6 and amrinone, we conclude that C6 produces its effect via potentiation of cAMP-dependent signaling system and possibly by inhibition of PDE3 activity.     

Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum

Background:Flagellated protozoan of the genus Leishmania is the causative agent of vector-borne parasitic diseases of leishmaniasis. Since the production of recombinant pharmaceutical proteins requires the cultivation of host cells in a serum-free medium, the elimination of FBS can improve the possibility of large-scale culture of Leishmania parasite. In the current study, we aimed at evaluating a new serum-free medium in Leishmania parasite culture for future live Leishmania vaccine purposes. Methods:Recombinant L. tarentolae secreting PpSP15-EGFP and wild type L. major were cultured in serum-free (CSFM) and serum-supplemented medium. The growth rate, protein expression, and infectivity of cultured parasites in both conditions was then evaluated and compared. Results:Diff-Quick staining and epi-fluorescence microscopy examination displayed the typical morphology of L. major and L. tarentolae-PpSP15-EGFP promastigote grown in CSFM medium. The amount of EGFP expression was similar in CSMF medium compared to M199 supplemented with 5% FBS in flow cytometry analysis of L. tarentolae-PpSP15-EGFP parasite. Also, a similar profile of PpSP15-EGFP proteins was recognized in Western blot analysis of L. tarentolae-PpSP15-EGFP cultured in CSMF and the serum-supplemented medium. Footpad swelling and parasite load measurements showed the ability of CSFM medium to support the L. major infectivity in BALB/C mice. Conclusion:This study demonstrated that CSFM can be a promising substitute for FBS supplemented medium in parasite culture for live vaccination purposes. Key Words: Growth rate, L. major, L. tarentolae, Serum-free medium, PpSP15-EGFP protein     

Effects of different carbon sources and dietary protein levels in a biofloc system on growth performance,immune response against white spot syndrome virus infection and cathepsin L gene expression of Litopenaeus vannamei

A 5‐week study was performed to evaluate the effect of spoilage date extract (SDE) as the biofloc carbon source on Litopenaeus vannamei (5.4 ± 0.3 g) performance. The two levels of dietary protein (15% and 25% crude protein) and two carbohydrate sources (molasses‐M and SDE‐P) were tested including: M15, M25, P15 and P25. The minimum (0.2 ± 0.0 mg/L) and the maximum (0.5 ± 0.0 mg/L) of total ammonia nitrogen were observed in the P15 and M25 groups respectively. The highest protein efficiency ratio (6.1 ± 0.3) and protein productive value (112.3 ± 5.8%) were found in the P15 group (p < 0.05). No significant difference was found between biofloc treatments in the expression of cathepsin L gene in hepatopancreas (p > 0.05). The number of total haemocyte count (THC), semigranular cells (SGC) and granular cells (GC) of shrimp in SDE‐based biofloc treatments was relatively higher than those in molasses‐based biofloc treatments. Following the white spot syndrome virus (WSSV) challenge, a significant decrease in THC, SGC, GC and hyaline cell values was observed in all treatments (p = 0.001). Plasma biochemical parameters were significantly influenced by dietary protein levels, biofloc carbon sources as well as WSSV challenge test. In conclusion, SDE successfully could be used as an alternative carbon source for establishing a biofloc system in L. vannamei production.     

Biochemical and Biological Evaluation of an L-Asparaginase from Isolated Escherichia coli MF-107 as an Anti-Tumor Enzyme on MCF7 Cell Line

Background: One of the most widely used anticancer agents is microbial L-ASNase. Herein, we assessed the biochemical and biological properties of an isolated L-ASNase from a Gram-negative bacteria strain, Escherichia coli MF-107. Methods:Using garden asparagus, we obtained several bacterial isolates. These strains were further screened for L-ASNase activity. A promising bacterial isolate was selected for L-ASNase production and subsequent purification. The molecular weight of purified L-ASNase was determined. The MTT assay was applied to assess the cytotoxic effect of the purified enzyme. Also, for caspase activity determination and the apoptotic effect of purified enzyme on in cells, we conducted a real-time PCR method. Results:The molecular weight of the enzyme was approximately 37 kDa. In the pH range of 7.5 to 8, the enzyme had considerable stability. At 35 °C, the purified L-ASNase optimum activity was recorded. The cytotoxic effect of the enzyme on treated cells was dose-dependent with an IC₅₀ value of 5.7 IU/ml. The Bax gene expression considerably raised by 5.75-fold (p < 0.001) upon L-ASNase treatment. On the other hand, the anti-apoptotic Bcl-2 gene expression showed a 2.63-fold increase compared to the control (p < 0.05). It was detected that the mRNA levels of caspase-3 and p53 were considerably upregulated (5.93 and 1.85-fold, respectively). We did not find any alternation in the caspase-8 activity of the treated cells compared to untreated cells. Conclusion:In this research, the proliferation of the breast cancer cells remarkably inhibited via the cytotoxic effect of isolated L-ASNase from microbial sources.Key Words: Apoptosis, Breast cancer, Escherichia coli, MCF7 cell line