Antimicrobial and Antibiofilm Activities of the Fungal Metabolites Isolated from the Marine Endophytes Epicoccum nigrum M13 and Alternaria alternata 13A

Epicotripeptin (1), a new cyclic tripeptide along with four known cyclic dipeptides (2–5) and one acetamide derivative (6) were isolated from seagrass-associated endophytic fungus Epicoccum nigrum M13 recovered from the Red Sea. Additionally, two new compounds, cyclodidepsipeptide phragamide A (7) and trioxobutanamide derivative phragamide B (8), together with eight known compounds (9–16), were isolated from plant-derived endophyte Alternaria alternata 13A collected from a saline lake of Wadi El Natrun depression in the Sahara Desert. The structures of the isolated compounds were determined based on the 1D and 2D NMR spectroscopic data, HRESIMS data, and a comparison with the reported literature. The absolute configurations of 1 and 7 were established by advanced Marfey’s and Mosher’s ester analyses. The antimicrobial screening indicated that seven of the tested compounds exhibited considerable (MIC range of 2.5–5 µg/mL) to moderate (10–20 µg/mL) antibacterial effect against the tested Gram-positive strains and moderate to weak (10–30 µg/mL) antibacterial effect against Gram-negative strains. Most of the compounds exhibited weak or no activity against the tested Gram-negative strains. On the other hand, four of the tested compounds showed considerable antibiofilm effects against biofilm forming Gram-positive and Gram-negative strains.     

Isolation and characterization of potassium solubilizing bacteria in some Iranian soils

The ability of a few soil bacteria to transform unavailable forms of potassium (K) to an available form is an important feature in plant growth-promoting bacteria for increasing plant yields of high-K-demand crops. In this research, isolation, screening, and characterization of six isolates of K solubilizing bacteria (KSB) from some Iranian soils were carried out. The ability of all isolates were tested in three treatments including acid-leached soil, biotite, and muscovite by analyzing the soluble K content after 5 days of incubation at 28 ± 2°C. Identification and phylogenetic analyses were also carried out by morphological, biochemical, and 16S rDNA analyses. Among the six efficient isolates, five isolates belonged to Bacillus megaterium (JK3, JK4, JK5, JK6, and JK7), while isolate JK2 belonged to Arthrobacter sp. The soluble K contents in all isolated-treatments were significantly (p < 0.01) higher than the contents in nonbacteria treatment. Herein, isolate JK2 had lower potential for K solubilization (910 mg kg^?1) compared with other isolates in acid-leached soils. The six bacterial strains showed higher solubilized K in biotite treatment than other two treatments. Overall, it can be concluded that the isolates belong to B. megaterium are the most efficient KSB under in vitro condition.     

Gill ATPase activities of silver perch, Bidyanus bidyanus (Mitchell), and golden perch, Macquaria ambigua (Richardson): Effects of environmental salt and ammonia

The Australian freshwater fish, silver and golden perch, are increasingly being used for aquaculture. Addition of salt to water is commonly used in commercial aquaculture to reduce stress attributed to high ammonia concentrations. The activities in gill homogenates of ouabain-sensitive Na⁺/K⁺-ATPase and NEM-sensitive ATPases (as a measure of H⁺-ATPases) of silver and golden perch were measured after maintaining the fish in water containing different salt and ammonia concentrations. Six treatments were applied in a 2 × 3 factorial design: two salt treatments, low salt (LS) of 2.5 g l^− 1 and high salt (HS) 5 g l^− 1, and three ammonia treatments, no added ammonia (NA), low ammonia (LA), 3 mg total ammonia nitrogen (TAN) l^− 1 and high ammonia (HA), 5 mg TAN l^− 1. In both species, activity of Na⁺/K⁺-ATPase was lowest in fish kept in the LSNA treatment (7.4 ± 0.4 μmol Pi mg protein^− 1 h^− 1 for silver perch and 3.1 ± 0.6 for golden perch) and highest in the HSHA treatment (15.2 ± 1.0 μmol Pi mg^− 1 protein h^− 1 for silver and 8.4 ± 1.2 for golden perch). In both species there was a significant increase (P < 0.001) in Na⁺/K⁺-ATPase activity with increase in salt concentration and with an increase in ammonia concentrations. A significant interaction (P < 0.036) between salt and ammonia on Na⁺/K⁺-ATPase activity was observed in silver but not in golden perch. In contrast, the lowest activity for NEM-sensitive ATPase was observed in the HSNA treatment (1.0 ± 0.2 μmol Pi mg^− 1 protein h^− 1 for silver and 1.5 ± 0.4 for golden perch) and highest in LSHA treatment (2.9 ± 0.4 μmol Pi mg^− 1 protein h^− 1 for silver and 3.6 ± 1.2 for golden perch). In both species there was a significant decrease in NEM-sensitive ATPase activity with increase in salt concentration and an increase in activity with increase in ammonia (P < 0.003). In silver perch, a significant interaction between the treatments was observed (P < 0.02). The results suggest that in these species of freshwater fish the Na⁺/K⁺-ATPase has a role in salt and ammonia homeostasis and that the NEM-sensitive ATPases are more active in fish kept in water with a lower salt content. It is possible that the increase in ammonia resistance when salt is added to the environmental water in commercial aquaculture systems may be due to the effects of salt on gill Na⁺/K⁺-ATPase activity rather than the NEM-sensitive ATPases.     

Effects of the application of ‘cash’ on the growth,fruit yield,and nutrient status of tomato (Solanum lycopersicum L.) grown in reclaimed saline soil

Summary

The effects of ‘cash’ [a novel 2:5:1 (w/w/w) mixture of calcium sulphate, ground sunflower heads, and humic acid] as a soil amendment on the growth, fruit yield, and leaf nutrient status of tomato (Solanum lycopersicum L.) grown on reclaimed saline soil (EC = 9.4 dS m^–1) were investigated. A glasshouse experiment was performed in a completely randomised design with six treatments (0, 10, 20, 30, 40, or 50 g ‘cash’ kg^–1 soil) each with four replicates. The results indicated that ‘cash’ increased both the shoot dry weight (DW) plant^–1 and the root DW plant^–1, the free proline contents of leaves, and leaf chlorophyll contents. The use of ‘cash’ also increased the number of fruit plant^–1, fruit yield pot^–1, and fruit vitamin C contents, although total soluble solids (TSS) contents were not affected. The contents of nitrogen (N), potassium (K), and calcium (Ca), and the Ca:Na ratios of leaf tissues increased with all application rates of ‘cash’. Leaf phosphorus (P) contents showed no response to any applied level of ‘cash’. Leaf sodium (Na) contents declined gradually with an increase in the level of ‘cash’ applied to the soil. We concluded that ‘cash’ has a pronounced positive effect on the growth and fruit yield of tomato plants grown in reclaimed saline soil. ‘Cash’ therefore has the potential to be used as a soil amendment for vegetable crops such as tomato to overcome the adverse effects of salinity in newly-reclaimed soils.     

Effect of nanoclay and magnesium hydroxide on some properties of HDPE/wheat straw composites

Since natural fiber/polymer composites are increasingly used, the development of safe and environmental friendly flame retarding bio-based composites is of great importance. But this issue must maintain the mechanical performance of these composites. To study these objectives, four levels of magnesium hydroxide Mg(OH)₂ of (0, 10, 20, 30 phc) and two levels of nanoclay (0, 3 phc) were considered and incorporated into HDPE/wheat straw composites. Maleic anhydride grafted polyethylene (PE-g-MA) was also used as a compatibilizer at constant content. The samples were prepared by melt compounding and injection molding processes, respectively. The some properties of samples including burning rate and mechanical properties (tensile and impact strengths) were tested based on the ASTM standard. The results showed that the burning rate of samples decreased with increasing the nanoclay and Mg(OH)₂ content. The tensile and impact strengths showed a marginal reduction by adding Mg(OH)₂ from 10 phc to 30 phc and the tensile modulus and impact strength revealed an increase by increasing the amount of nanoclay up to 3 phc. Generally, these results confirmed that the fire retarding and mechanical properties of HDPE/wheat straw composites could be significantly improved with an appropriate combination of the nanoclay and Mg(OH)₂ in the composites.     

Hypocalcemic seizures in breastfed infants with rickets secondary to severe maternal vitamin D deficiency

This study was done to evaluate if nursing mothers of infants with rickets have vitamin D deficiency, and to evaluate the relationship between maternal vitamin D levels with hypocalcemic seizures in infants with rickets. We selected a cohort of breastfed infants with rickets. Infants were included in this study if they were breastfed and presented with any of the following clinical criteria: delayed motor milestones or delayed teething, were found to have specific rachitic bony signs, or presented with hypocalcemic seizures. We checked serum calcium (Ca), phosphorus (P), alkaline phosphatase, 25 hydroxy vitamin D [25(OH)D] and parathyroid hormone (PTH) levels in both infants and their mothers. Out of 32 children who met the clinical criteria for rickets, 23 (72%) had vitamin D level less than 20 ng mL(-1). Twenty two mothers (69%) had vitamin D deficiency (25 hydroxy vitamin D < 20 ng mL(-1)). Mothers of nine infants who presented with hypocalcemic seizures had severe vitamin D deficiency, (p = 0.005). We conclude that maternal vitamin D deficiency is common in nursing mothers of infants diagnosed with rickets. Invariably mothers of infants presenting with hypocalcemic seizures have severe vitamin D deficiency. Hypocalcemic seizures in infants secondary to maternal vitamin D deficiency might be prevented by supplementation of vitamin D.     

Extracts of Oliviera decumbens and Satureja khuzestanica as immunostimulants affect some innate immunity indices of Cyprinus carpio against Aeromonas hydrophila infection

In this study, the effect of extracts of two herbs (Oliviera decumbens and Satureja khuzestanica) on immune response of carp was investigated. At the beginning of experiment, fish were divided to two groups including vaccinated (using vaccine developed against A. hydrophila) and non‐vaccinated. Fish in both groups were fed diets containing O. decumbens and S. khuzestanica and combination of two herbal extracts for 5 weeks. Control fish (negative control) and fish vaccinated only (positive control) were fed basal diets without supplements of herbal extracts. Lysozyme activity, antibody titre, complement activity and bactericidal activity in serum were measured. After 5 weeks feeding, fish were infected with A. hydrophila and mortalities were recorded. In both experimental groups, no significant differences were found in terms of alternative haemolytic complement (ACH50) activity and antibody titres of Serum. In non‐vaccinated fish group, lysozyme and bactericidal activity of fish fed S. khuzestanica or combination of O. decumbens and S. khuzestanica was higher compared with control and other experimental treatments. In vaccinated fish group, the lysozyme and bactericidal activity was not significant in all treatments compared with control group. The results of this study showed that feeding non‐vaccinated and vaccinated carp with only S. khuzestanica or in combination with O. decumbens enhance only some immunity indices including lysozyme and bactericidal activity in non‐vaccinated fish and these extracts have no immunological stimulatory role on vaccinated individuals.     

A comparative histopathological, immunohistochemical and nested-PCR study for diagnosis of infectious pancreatic necrosis in the naturally infected cultured rainbow trout (Oncorhynchus mykiss)

To confirm the diagnosis of the infectious pancreatic necrosis, the pancreas, liver, kidney and spleen specimens from 140 rainbow trout (Oncorhynchus mykiss) of weight ranges from 15 to 250?g were processed for routine histopathological and immunohistochemical (IHC) studies together with a nested-PCR using primers that amplify a 164-bp product. Of the 140 fish samples, 37 (26.4%) had a final diagnosis of IPN on the basis of hematoxylin and eosin staining (H&E), while 39/45 (86.6%) were positive by IHC method. With the nested-PCR, 59/140 (42.1%) fish samples were positive. The IHC and nested-PCR showed higher prevalence than histopathology (P?<?0.05). Based on the nested-PCR, as the fish weight increased, IPN positive results decreased (P?<?0.05). However, IHC method detected IPNV constantly in these weight ranges. Only IPNV serotype Sp was identified by nucleotide sequencing and immunohistochemistry, and antiserum to IPNV serotype Ab and IHNV showed negative results in IHC. This is the first comparative diagnostic study of IPN at different weight ranges in cultured salmonids.     

Ambivalent effects of atorvastatin on angiogenesis, epidermal cell proliferation and tumorgenesis in animal models

Background: A growing body of preclinical data indicates that statins may possess antineoplastic properties; however, some studies have raised the possibility that statins may also have carcinogenic potential. Methods: An air pouch model was used for angiogenesis. Single or multiple applications of croton oil on the back of Swiss albino mice with or without initiation by dimethylbenz(a)antheracene (DMBA) were used to evaluate the skin tumorgenesis, ultrastructural and histological alterations. Results: Atorvastatin (orally, 10 mg/kg/day) produced a significant (P<0.05) reduction in angiogenesis. Concurrent administration of mevalonate reversed the anti-angiogenic effect of atorvastatin. However, local injection of atorvastatin (200 µg) into the pouches induced a significant (P<0.5) increase in angiogenesis that was not reversed by co-administration of mevalonate. The disturbance of cell polarity, inflammatory response, thickness of epidermal layer, and mitotic index induced by croton oil were inhibited markedly and dose-dependently (P<0.001) by pre-treatment with atorvastatin. In spite of the strong anti-inflammatory and anti-proliferative effects of atorvastatin on epidermal cell proliferation, it was identified that the same doses of atorvastatin in DMBA-initiated and croton oil-promoted skin tumorgenesis in mice increased the incidence of tumors and their conversion into malignant carcinoma. Conclusion: The reasons for these discrepancies remain unclear, and could be related to ambivalent effects of atorvastatin on angiogenesis or to specific differences in the experimental conditions. It is suggested that the pro-angiogenic effect of the drug, which could be responsible for promotion of skin tumors, is independent of the 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase inhibition that can be mediated directly by atorvastatin.Key Words: Atorvastatin, Angiogenesis, Cell Proliferation, Cancer     

Spermiation time affect the milt quality indices of the Russian sturgeon,Acipenser gueldenstaedtii,Brandt & Ratzeburg, 1833

In this study, the effects of spermiation time are investigated on milt quality of Russian sturgeon over the course of the spawning season. The milt samples were collected from three broodstock batches at three time points including: the beginning, middle and at the end of the spawning season. According to the results, the milt quality parameters including pH, sperm density, spermatocrit, duration of sperm motility and percentage of sperm motility were significantly low in the beginning and end of season than middle of season. The values of milt quality parameters in the middle of season were as follows: (motility percentage: 69.6 ± 3.5, motility duration: 460.3 ± 37.2 s, sperm density: 8.7 ± 0.4 × 10⁹, milt volume: 86.3 ± 8.1 and milt pH: 8.3 ± 0.15). Significant positive correlations were also found between milt pH and sperm motility as well as between sperm density and spermatocrit. In conclusion, our study showed that the middle of season is the best time for collection of milt with appropriate quality in Russian sturgeon. Selection of milt with good quality is necessary aim to cryopreservation of spermatozoa in endangered fish species including Russian sturgeon.