Production of Induced Secondary Metabolites by a Co-Culture of Sponge-Associated Actinomycetes,Actinokineospora sp. EG49 and Nocardiopsis sp. RV163

Two sponge-derived actinomycetes, Actinokineospora sp. EG49 and Nocardiopsis sp. RV163, were grown in co-culture and the presence of induced metabolites monitored by ¹H NMR. Ten known compounds, including angucycline, diketopiperazine and β-carboline derivatives 1–10, were isolated from the EtOAc extracts of Actinokineospora sp. EG49 and Nocardiopsis sp. RV163. Co-cultivation of Actinokineospora sp. EG49 and Nocardiopsis sp. RV163 induced the biosynthesis of three natural products that were not detected in the single culture of either microorganism, namely N-(2-hydroxyphenyl)-acetamide (11), 1,6-dihydroxyphenazine (12) and 5a,6,11a,12-tetrahydro-5a,11a-dimethyl[1,4]benzoxazino[3,2-b][1,4]benzoxazine (13a). When tested for biological activity against a range of bacteria and parasites, only the phenazine 12 was active against Bacillus sp. P25, Trypanosoma brucei and interestingly, against Actinokineospora sp. EG49. These findings highlight the co-cultivation approach as an effective strategy to access the bioactive secondary metabolites hidden in the genomes of marine actinomycetes.     

Effect of alpha-interferon and alpha-tocopherol in reversing hepatic cirrhosis in rats

The aim of this study was to assess the effects of alpha-interferon and alpha-tocopherol (vitamin E), or a combination of both, in reversing hepatic fibrosis following the induction of cirrhosis using thioacetamide by histological and biochemical analysis. Fifty male Wistar rats were used in this study. The animals were divided equally into five groups. Animals in group I were used as controls. The remaining animals (groups II-V) were provided with 0.5 g/L of thioacetamide in order to induce liver cirrhosis. Group II animals were used as the cirrhotic control. Animals of groups III, IV and V were given alpha-interferon, alpha-tocopherol and interferon together with alpha-tocopherol, respectively, for 30 days. After 30 days the animals were killed and following gross morphological examination of the liver, the hepatic tissues were processed for histological analysis and the serum was used for liver function tests. Morphological analysis showed a decrease in the number of nodules on the surface of the liver in both interferon- as well as vitamin E-treated cirrhotic rats. Histopathological analysis showed that the abnormalities of the cirrhotic liver were partially reversed and liver function tests showed an overall improvement following treatment of animals of groups III, IV and V. Combination therapy using both interferon and alpha-tocopherol did not have any substantial effect on the rats compared with that when they were given separately. These findings suggest that alpha-interferon and alpha-tocopherol may have therapeutic value in reversing liver cirrhosis.     

Genotypic analysis of virulence genes and antimicrobial profile of diarrheagenic <Emphasis Type="Italic">Escherichia coli</Emphasis> isolated from diseased lambs in Iran

The aim of the present study was to determine the analysis of virulence genes and antimicrobial profile of diarrheagenic Escherichia coli isolated from diseased lambs. Two hundred ninety E. coli isolates were recovered from 300 rectal swabs of diarrheic lambs and were confirmed by biochemical tests. The pathotype determination was done according to the presence of genes including f5, f41, LTI, STI, bfp, ipaH, stx ₁ , stx ₂ , eae, ehlyA, cnf ₁ , cnf ₂ , cdIII, cdIV, and f17 by PCR method. Sixty-six isolates (23.72%) possessed the STI gene and categorized into entrotoxigenic E. coli (ETEC). Nine isolates (3.1%) and five isolates (1.72%) were positive for the cnf1 and cnf2 genes which categorized into necrotoxic E. coli (NTEC). Hundred and seventeen isolates (40.34%) harbored stx ₁ and/or stx ₂ and classified as Shiga toxin-producing E. coli (STEC). Thirteen isolates (4.48%) were assigned to atypical entropathogenic E. coli (aEPEC) and possessed eae gene. Two isolates (0.68%) were positive for ipaH gene and were assigned to entroinvasive E. coli (EIEC). Statistical analysis showed a specific association between eae gene and STEC pathotype (P?<?0.0001). The most prevalent resistance was observed against lincomycin (96.5%) and the lowest resistance was against kanamycine (56.02%), respectively. The high prevalence of STEC and ETEC indicates that diarrheic lambs represent an important reservoir for humans. ETEC may play an important role for frequent occurrence of diarrhea in lambs observed in this region. Due to high antibiotic resistance, appropriate control should be implemented in veterinary medicine to curb the development of novel resistant isolates.     

A review of the contemporary knowledge of bovine tuberculosis and government policy in Iran

In 1931 Carpantier reported bovine TB (BTB) in Iranian cattle. Some eighty years on with a national test-and-slaughter programme in place for over four decades, the efforts to vanquish Mycobacterium bovis (M. bovis) infection in cattle have been in vain as the vast majority of the 30 Iranian provinces still have reports of BTB in their cattle herds every year. This paper reviews the present epidemiology of BTB in Iran and in the region and evaluates the success of government policy in controlling this disease.     

Brain selenium accumulation in rat pups of selenium supplemented mothers

Selenium is essential for normal mammalian development. Being a component of antioxidant enzyme, glutathione peroxidase, it plays a major role in protecting the cells from free radical damage. The level of glutathione peroxidase was directly related to the amount of selenium present in various tissues and organs. A decrease in selenium leads to various pathological changes in humans as well as in various laboratory animals. The aim of the present study was to understand whether there is an increase in the level of selenium in different brain regions of rat pups whose mothers were supplemented with selenium, either 2 or 4 mg/l of their drinking water throughout the period of their pregnancy. There was a significant increase in the level of selenium in the cerebellum, cortex and hypothalamic and hippocampal tissues of selenium supplemented mothers as compared with those of non-supplemented mothers. The brain stem of these animals did not show any significant difference in the level of selenium. Furthermore, the differences in the level of selenium between the rat pups of 2 mg/l selenium supplemented mothers and 4 mg/l selenium supplemented mothers were not statistically significant. These studies suggest that supplementation of selenium to mothers during the period of their pregnancy can selectively increase the level of this trace element in different brain regions. Further studies are necessary to understand the significance of selective accumulation of selenium in specific brain regions on brain development and function.     

First insight into genetic diversity and population structure of the Caucasian wild apple (<Emphasis Type="Italic">Malus orientalis</Emphasis> Uglitzk.) in the Hyrcanian forest (Iran) and its resistance to apple scab and powdery mildew

Investigating population genetic structure and diversity, and resistance to pathogens in crop wild relatives are key steps to assess appropriate conservation and breeding programs. The Caucasian wild apple (Malus orientalis Uglitzk.) is an emblematic fruit tree of the Hyrcanian forest and is supposed to be a contributor to the cultivated apple genome (Malus domestica Borkh. Yet, no study has investigated its population structure, diversity and susceptibility to the two main pathogens of apples, the apple scab (Venturia inaqualis) and the powdery mildew (Podosphaera leucotricha). Here, we investigated population genetic structure and diversity of M. orientalis in the Hyrcanian forest as a mean to identifying future targeted populations for apple conservation and breeding programs. We genotyped using multilocus microsatellite 100 M. orientalis trees sampled in 14 sites. These trees were also screened for presence/absence of six (Rvi6, Vr, Rvi4, Rvi15, Rvi5 and Rvi11) and three (Pl-1, Pl-w, Pl-d) resistance genes to the apple scab and the powdery mildew respectively. Our results showed significant but weak between-site genetic differentiation and isolation by distance pattern suggesting substantial historical gene flow for M. orientalis in this area. We also detected a West-Eastern genetic structure across the Hyrcanian forest with five main populations showing admixture. We also showed a high diversity of resistance genes to apple scab across sites; in contrast, we only found one resistance gene to powdery mildew. These results are a first glimpse to settle wild apple conservation programs in Iran and pinpoint Iranian wild apple populations as an untapped source for apple breeding.     

Combined effects of plant growth‐promoting rhizobacteria and genistein on nitrogen fixation in soybean at suboptimal root zone temperatures

Application of plant growth‐promoting rhizobacteria (PGPR) or the plant to bacteria signal molecule genistein has been shown to increase nodulation and nitrogen (N) fixation by soybean [Glycine max (L.) Merr.] over a range of root zone temperatures (RZTs) and, specifically, off‐sets at least some of the ill‐effects of low RZTs. Two sets of controlled‐environment experiments, one on a growth bench and the other in a greenhouse, were conducted to examine the combined ability of both PGPR and genistein to reduce the negative effects of low RZT on soybean nodulation and N fixation. Each of two the PGPR strains, Serratia proteamaculans 1–102 and Serratia liquefaciens 2–68 were co‐inoculated with Bradyrhizobium japonicum USDA 110 preincubated with 17.5 (somewhat inhibitory), and 15°C (very inhibitory). At RZTs of 25 and 17.5°C PGPR strains and genistein in combination increased the number of nodules and the amount of Nn fixed. The most stimulatory effect was observed at 17.5°C for the combination: S. proteamaculans 1–102 plus B. japonicum USDA 110 pre‐incubated in 15 μM genistein under greenhouse conditions. For most treatment combinations the stimulatory effects of PGPR and genistein were additive at RZTs of 17.5 and 25°C. Surprisingly, the combination of these two factors resulted in antagonism at the very inhibitory RZT of 15°C. The results suggest that the negative effects of certain low RZTs could be more effectively off‐set by combined treatments of PGPR plus geneistin pre‐incubation of rhizobial cultures than by their individual treatment.     

Root and Rhizosphere Colonization of Soybean [Glycine max (L.) Merr.] by Plant-Growth-Promoting Rhizobacteria at Low Root Zone Temperatures and under Short-Season Conditions

Co-inoculation of plant-growth-promoting rhizobacteria (PGPR) with B. japonicum has been shown to increase soybean [ Glycine max (L.) Merr.] nodulation, nitrogen fixation, growth and physiological activity at suboptimal root zone temperatures (RZTs). We studied the survival and growth of seven PGPR inoculated on soybean in a sterile rooting medium at three RZTs (25, 17.5 and 15 °C) on a growth bench. The survival of the two most promising strains ( Serratia liquefaciens 2-68 and S. proteamaculans 1-102) was studied under field conditions in methyl bromide fumigated and non-fumigated soils. In general, population densities varied with temperature. PGPR strains generally colonized the rhizosphere and root surface effeciently at higher RZTs; however, S. proteamaculans 1-102 colonized best at a low RZT (15 °C). The population of PGPR applied to the rhizosphere either with or without addition of B. japonicum increased over time in fumigated soil as compared to non-fumigated soil, indicating that the PGPR survive and proliferate better under fumigated conditions. S. liquefaciens 2-68 had higher population densities both on the root and in the rhizosphere, demonstrating, their ability to colonize under short-season conditions. The possible interactions between the two cultivars (Maple Glen and AC Bravor) and the PGPR were generally not significant, despite observations that growth and yield of AC Bravor respond more strongly to PGPR inoculation.     

Overcoming seed dormancy of mooseer (Allium hirtifolium) through cold stratification, gibberellic acid, and acid scarification

Mooseer(Allium hirtifolium Boiss.) is bulbous perennial herb widely used in pharmaceutical and food industry in Iran.We studied germination of mooseer seeds in two separate experiments.In the first experiment,we evaluated four treatments:sulfuric acid scarification,sandpaper scarification,cold stratification,and gibberellic acid(GA 3) application.In the second experiment,we evaluated combinations of these treatments.All treatments in the first experiment had no effect on seed germination,suggesting that mooseer seeds have physical and physiological dormancy.In the second experiment,the highest germination percentage(86.6%) was observed after five minutes scarification with sulfuric acid(75% v/v),followed by 60 days of cold stratification.Duration of sulfuric acid scarification(5,10,and 20 min) did not affect germination rates,but increasing duration of cold stratification(from 15 to 60 days),increased germination from 28.3% to 86.6%.Our study showed that mooseer seeds have both physical and physiological dormancy.     

EFFECTS OF SALINITY AND SOIL ZINC APPLICATION ON GROWTH AND CHEMICAL COMPOSITION OF PISTACHIO SEEDLINGS

The effects of four salinity levels [0, 1000, 2000, and 3000 mg sodium chloride (NaCl) kg^?1 soil] and three zinc (Zn) levels [0, 5, and 10 mg kg^?1 soil as zinc sulfate (ZnSO₄.7 H₂O)] on growth and chemical composition of pistachio seedlings (Pistacia vera L.) cv. ‘Badami’ were studied in a calcareous soil under greenhouse conditions in a completely randomized design with three replications. After 26 weeks, the dry weights of leaves, stems and roots were measured and the total leaf area determined. Salinity decreased leaf, stem, and root dry weights and leaf area, while this effect diminished with increasing Zn levels. Zn fertilization increased leaf, stem and root Zn concentrations, leaf potassium (K) concentration, and stem and root sodium (Na) concentrations, while decreased leaf Na concentration, and stem and root K concentrations. Salinity stress decreased leaf, stem, and root Zn concentrations, and leaf K concentration, while salinity increased leaf, stem and root Na concentrations, and stem and root K concentrations. Proline accumulation increased with increasing salinity levels, whereas the reverse trend was observed for reducing sugar contents. Zn application decreased proline concentration but increased reducing sugar contents. These changes might have alleviated the adverse effects of salinity stress.