Morphological and phylogenetic diversity of thermophilic cyanobacteria in Algerian hot springs

Geothermal springs in Algeria have been known since the Roman Empire. They mainly locate in Eastern Algeria and are inhabited by thermophilic organisms, which include cyanobacteria forming mats and concretions. In this work, we have investigated the cyanobacterial diversity of these springs. Cyanobacteria were collected from water, concretions and mats in nine hot springs with water temperatures ranging from 39 to 93 °C. Samples were collected for isolation in culture, microscopic morphological examination, and molecular diversity analysis based on 16S rRNA gene sequences. Nineteen different cyanobacterial morphotypes were identified, the most abundant of which were three species of Leptolyngbya, accompanied by members of the genera Gloeocapsa, Gloeocapsopsis, Stigonema, Fischerella, Synechocystis, Microcoleus, Cyanobacterium, Chroococcus and Geitlerinema. Molecular diversity analyses were in good general agreement with classical identification and allowed the detection of additional species in three springs with temperatures higher than 50 °C. They corresponded to a Synechococcus clade and to relatives of the intracellularly calcifying Candidatus Gloeomargarita lithophora. The hottest springs were dominated by members of Leptolyngbya, Synechococcus-like cyanobacteria and Gloeomargarita, whereas Oscillatoriales other than Leptolyngbya, Chroococcales and Stigonematales dominated lower temperature springs. The isolation of some of these strains sets the ground for future studies on the biology of thermophilic cyanobacteria.     

Production of plant growth substances by rhizosphere mycoflora of broad bean and cotton

The culture filtrates of the rhizosphere fungi of broad bean (Vicia faba L.) and cotton (Gossypium barbadense L.) were analysed for the presence of plant growth substances of auxin and gibberellins nature. Bioassay test and chromatographic analysis indicated that these fungi, each synthesized different auxins in their culture medium. These auxins were indole compounds. Similarly the rhizosphere fungi produced in their culture medium some gibberellins and gibberellin-like substances.     

Diversity and antibacterial activities of culturable fungi associated with coral Porites pukoensis

The diversity of coral associated fungi is not enough understood, especially for scleractinian corals. Members of Porites are common and dominant species of scleractinian corals. To date, the fungal communities associated with coral Porites pukoensis have been not reported. In this paper, the diversity and activity of coral associated fungi in P. pukoensis were explored, 23 fungal strains were isolated, belonging to 10 genera and Aspergillus sp. (30.4 %) was predominant fungal genera. The sequence of isolate C1-23 in GenBank was only 90 % similarity to the most closely related sequences. It is concluded that rich fungal symbionts are attached to P. pukoensis, the rate of isolates with antibacterial activity was up to 30 %, particularly some isolates showed stronger bioactivities to gram-negative bacteria. It is included that the diversity of coral associated fungi in P. pukoensis is abundant and its activity is obviously. So the activities of fungi in P. pukoensis were deserved for further study.     

The survival of microfungi in the nests of Tree-Sparrow [Passer montanus L.] in the nest-boxes over the winter season

At the mycological examination of eleven nest-boxes of the Tree-Sparrow (Passer montanus L.) after the winter season a significant increase of the total number of fungi was ascertained in comparison with the investigation of the same boxes in the previous summer; especially the cellulolytic fungi increased (generaPenicillium, Scopulariopsis, Chrysosporium et al.) but also the potentially zoopathogenic (ascertainedAspergillus fumigatus, Candida albicans, Mucor pusillus et al.) and toxinogenic fungi (e.g.Asp. flavus); a moderate-statistically insignificant-decrease in the number of isolates and species of keratinolytic fungi took place (in spring foundArthroderma ciferrii, A. quadrifidum andAphanoascus fulvescens). In some fungi not only survival but also colonization of the substrate in the winter and early-spring period were proved. As main factors conditioning the survival of fungi in the nest-boxes over the winter were ascertained the composition of the substrate, its temperature and humidity. The limiting abiotic factor is before all the low temperature, inhibiting the development of nonpsychorphilic fungi. On the base of various changes of the physical and chemical conditions of the environment an attempt of explaining the dynamics of some ascertained fungi in accordance with literary statements is presented. The proved overwintering of some potentially pathogenic fungi in the microhabitat supplies to the studies of this kind an epidemiologic and epizootologic importance.     

Effects of Icotinib on Advanced Non-Small Cell Lung Cancer with Different EGFR Phenotypes

Icotinib is the first oral epidermal growth factor receptor (EGFR) tyrosine kinase receptor inhibitor, which has been proven to exert significant inhibitory effects on non-small cell lung cancer in vitro. Clinical evidence has showed that the efficacy of Icotinib on retreating advanced non-small cell lung cancer is comparable to Gefitinib. However, different phenotypes of EGFR can affect the therapeutic outcomes of EGFR tyrosine kinase receptor inhibitor. Therefore, our study focused on efficacy and safety of Icotinib in patients with advanced non-small cell lung cancer of different EGPR phenotypes. Clinical data of patients with advanced non-small cell lung cancer who received Icotinib treatment from August, 2011 to May, 2013 were retrospectively analyzed. Kaplan–Meier analysis was used for survival analysis and comparison. 18 wild-type EGFR and 51 mutant type were found in a total of 69 patients. Objective response rate of patients with mutant type EGFR was 54.9 % and disease control rate was 86.3 %. Objective response rate of wild-type patients was 11.1 % (P = 0.0013 vs mutant type), disease control rate was 50.0 % (P = 0.0017). Median progression-free survival (PFS) of mutant type and wild-type patients were 9.7 and 2.6 months, respectively (P < 0.001). Median PFS of exon 19 mutated mutant patients was 11.3 months, mean PFS of exon 21 L858R mutated mutant patients was 8.7 months (P = 0.3145). Median overall survival (OS) of EGFR mutated patients had not reached. OS time of 13 wild-type patients was 12.9 months (P < 0.001). The common adverse reactions of Icotinib included rash, diarrhea, itching skin with occurrence rates of 24.6 % (17/69), 13.0 % (9/69), and 11.6 % (8/69), respectively. Most adverse reactions were grade I–II. Icotinib has great efficacy in EGFR mutated patients, making it an optimal regimen to treat EGFR mutated patients. Furthermore, most of adverse reactions associated with Icotinib treatment were tolerable.     

Determination of guanosine tetraphosphate (ppGpp) and adenosine pentaphosphate (pppApp) in various microorganisms by radioimmunoassay

The intracellular levels of ppGpp and pppApp in various microorganisms were determined by radioimmunoassay, with was of more accuracy and convenience than previous ³²P-labeling method. ppGpp was detected in bacteria, actinomycetes, yeasts and fungi. pppApp was found in non-spore forming bacteria such as the genera Pseudomonas and Escherichia, fungi and actinomycetes, but not in yeasts. In conclusion, ppGpp and pppApp are present in various prokaryotes and lower eukaryotes.     

Studies on the rhizosphere mycoflora of broad bean and cotton

Investigations were made on the effect of species of fungi isolated from therhizosphere of broad bean (Vicia faba Linn.) variety Giza I, and cotton (Gossypium barbadense Linn.) variety Giza 47, on plant growth. Broad bean rhizosphere fungi differently affected plant growth.Fusarium moniliforme, Penicillium martensii, Rhizopus stolonifer, andCladosporium sphaerospermum stimulated both root and shoot growth.Aspergillus niger andAlternaria tenuis have an inhibitory effect on plant growth. On the other hand, the rhizosphere fungi of cotton namely,Penicillium cyclopium, Aspergillus terreus, Cephalosporium sp.,Aspergillus niger, Cladosporium sphaerospermum, andFusarium oxysporum stimulated plant growth.     

Studies on the rhizosphere mycoflora of broad bean and cotton. Effect of fungal filtrate on plant growth

The dominating rhizosphere fungi of broad bean (Vicia faba Linn.) variety “Giza 1”, and cotton (Gossypium barbadense Linn.) variety “Giza 47”, were grown in liquid medium. After 10 days, filtrates were obtained and sterilized by filtration through sintered-glass filter. Plants were grown in sterile sand which was supplemented with nutrient solution. Every plant was irrigated with fungal filtrate, unconsumed medium, and water. The filtrates of the rhizosphere fungi of both broad bean and cotton stimulated plant growth.     

Prey range of the predatory ladybird Cryptolaemus montrouzieri

The prey range of Cryptolaemus montrouzieri was studied in the laboratory to investigate whether the mealybug destroyer can contribute to the suppression of other pest insects besides mealybugs and to assess its potential impact on non-mealybug populations as part of an environmental risk assessment for its use in biological control. Prey tested in these experiments were: tobacco aphid Myzus persicae nicotianae (Sulzer)(Hemiptera: Aphididae), pea aphid Acyrthosiphon pisum (Harris)(Hemiptera: Aphididae), tobacco whitefly Bemisia tabaci (Gennadius)(Hemiptera: Aleyrodidae), southern green stinkbug Nezara viridula (L.)(Hemiptera: Pentatomidae) eggs, western flower thrips Frankliniella occidentalis (Pergande)(Thysanoptera: Thripidae), two-spotted ladybird Adalia bipunctata (L.)(Coleoptera: Coccinellidae) eggs and eggs of the greater wax moth Galleria mellonella L. (Lepidoptera: Pyralidae). Larval survival was high to moderate when C. montrouzieri was provided with hemipteran prey and poor to zero when the ladybirds were provided with non-hemipteran prey. Females reared on M. persicae and A. pisum produced similar numbers of eggs as their counterparts fed the citrus mealybug Planococcus citri (Risso)(Hemiptera: Pseudococcidae), but fecundity was significantly lower when the ladybirds were reared on B. tabaci nymphs or on A. bipunctata eggs. Prey species that were found to be less suitable for immature development of C. montrouzieri could still be an adequate food source for reproduction and survival of adult ladybirds. For example, only 8 % of the predator larvae reached the adult stage when provided with A. bipunctata eggs, but females that had developed on eggs of the Mediterranean flour moth Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) and that were supplied with A. bipunctata eggs from adult emergence on, were only 35 % less fecund than females provided with mealybugs in their adult life. The results are discussed in relation to the development of a suitable methodology for prey/host range testing in the framework of an environmental risk assessment for arthropod natural enemies.     

Antimicrobial activity of some Indian plants

Antimicrobial activity of 105 Indian plant species was tested. Among them, 30 showed antibacterial activity; 20 of these exhibited antifungal action as well. Seeds ofCarum copticum, stem ofPinus longifolia, roots ofPlumbago zeylanica andSaussurea lappa, and rhizome ofAlpinia officinarum have considerable antifungal activity, especially against pathogenic fungi. Antibiotic activity against a wide variety of microorganisms—pathogenic and nonpathogenic gram-positive and gram-negative bacteria, yeast, and fungi—was also noted with leaves ofLawsonia inermis, roots ofPlumbago zeylanica, and fruits ofTamarindus indica,Terminalia belerica, andEmblica officinalis.     

Persistence of antibiotic-resistant and -sensitive Proteus mirabilis strains in the digestive tract of the housefly (Musca domestica) and green bottle flies (Calliphoridae)

Synanthropic flies have been implicated in the rapid dissemination of antibiotic-resistant bacteria and resistance determinants in the biosphere. These flies stably harbor a considerable number of bacteria that exhibit resistance to various antibiotics, but the mechanisms underlying this phenomenon remain unclear. In this study, we investigated the persistence of antibiotic-resistant bacteria in the digestive tract of houseflies and green bottle flies, using Proteus mirabilis as a model microorganism. One resistant strain carried the blaTEM and aphA1 genes, and another carried a plasmid containing qnrD gene. Quantitative PCR and 454 pyrosequencing were used to monitor the relative abundance of the Proteus strains, as well as potential changes in the overall structure of the whole bacterial community incurred by the artificial induction of Proteus cultures. Both antibiotic-resistant and -sensitive P. mirabilis strains persisted in the fly digestive tract for at least 3 days, and there was no significant difference in the relative abundance of resistant and sensitive strains despite the lower growth rate of resistant strains when cultured in vitro. Therefore, conditions in the fly digestive tract may allow resistant strains to survive the competition with sensitive strains in the absence of antibiotic selective pressure. The composition of the fly-associated bacterial community changed over time, but the contribution of the artificially introduced P. mirabilis strains to these changes was not clear. In order to explain these changes, it will be necessary to obtain more information about bacterial interspecies antagonism in the fly digestive tract.     

Culturable fungal endophytes in roots of Enkianthus campanulatus (Ericaceae)

Roots of plants in the genus Enkianthus, which belongs to the earliest diverging lineage in the Ericaceae, are commonly colonized by arbuscular mycorrhizal (AM) fungi. We documented the community of fungal root endophytes associated with Enkianthus species using a culture-based method for better understanding the members of root-colonizing fungi, except for AM fungi. Fungal isolates were successfully obtained from 610 out of 3,599 (16.9 %) root segments. Molecular analysis of fungal cultures based on ribosomal internal transcribed spacer (ITS) sequences yielded 63 operational taxonomical units (OTUs: 97 % sequence similarity cutoff) from 315 representative isolates. Further phylogenetic analysis showed that most (296 isolates) belonged to Ascomycota and were either members of Helotiales (Dermataceae, Hyaloscyphaceae, Phialocephala and Rhizoscyphus ericae aggregate), Oidiodendron, or other Pezizomycotina. Twenty-three out of 63 OTUs, which mainly consisted of Leotiomycetes, showed high similarities with reference sequences derived from roots of other ericaceous plants such as Rhododendron. The results indicated that Enkianthus houses variable root mycobionts including putative endophytic and mycorrhizal fungi in addition to AM fungi.     

Investigation of the internal bacterial flora of Eurygaster integriceps (Hemiptera: Scutelleridae) and pathogenicity of the flora members

Eurygaster integriceps (Hemiptera: Scutelleridae) is one of the most serious pest of wheat and other cereal crops throughout the Near and Middle East including Turkey. To contribute biological control of this pest, we investigated the culturable bacterial flora of E. integriceps and their virulence against it. We also tested different entomopathogenic fungi (Isaria fumosorosea ARSEF8356, Beauveria bassiana ARSEF8356, Metarhizium brunneum ARSEF8671 and Nomurae rileyi ARSEF1670) against this pest under laboratory conditions. Bacterial isolates were characterized based on morphological, biochemical, physiological and molecular characteristics (16S rRNA sequencing). The isolates were identified as Pantoea sp. (S1, S5, S7, S8, S10 and S11), P. agglomerans (S2, S3 and S4), Pseudomonas sp. S6 and Micrococcus luteus S9. The highest mortality within bacterial isolates was observed from Pantoea sp. S1, P. agglomerans S4 and Pantoea sp. S7 with 100%. Mortality of other bacterial species ranged from 33% to 88%. The highest mortality among the tested entomopathogenic fungi was obtained from Isaria fumosorosea ARSEF 8333 with 100%. Mortality of other fungi ranged from 33% to 50%. Consequently, Pantoea sp. S1, P. agglomerans S4, Pantoea sp. S7, Beauveria bassiana ARSEF 8356 and Isaria fumosorosea ARSEF 8356 seem to be promising candidates in the control of E. integriceps.     

Expression of genes in the 111.35–116.16 million bp fragment of chromosome 13 in brain of mice with different predisposition to hereditary catalepsy

Catalepsy is a pathological animal behavior that is usually associated with dysfunctions in the striatal pallidal system of the brain and can be caused by different reasons. It was previously demonstrated that hereditary catalepsy is linked to the 111.35–116.16 million bp fragment of chromosome 13 in mice. The level of mRNA content in 42 genes localized in this fragment was determined in the study. Two brain departments that are functionally associated with catalepsy (striatum and substantia nigra) were studied in mice from AKR line (resistant to catalepsy), cataleptic CBA line, and recombinant cataleptic AKR.CBA-D13Mit76 (D13) line. The latter was obtained by the transfer of indicated fragment of chromosome 13 from the CBA line to the genome of the AKR line. It was found that two genes (Ndufs4 and Ppap2a) in the striatum and ten genes (Esm1, Fst, Gm10735, Gm15322, Gm15323, Gm15324, Gm15325, Il6st, Il31ra, and Itga1) in the substantia nigra differ in the level of mRNA expression in AKR and D13 lines. The Mcidas gene mRNA level is lower in both structures in D13 line mice than in the AKR line. The expression of the Hspb3 and Mocs2 genes (that encode heat shock protein and molybdenum cofactor synthesis, respectively) is lower in the substantia nigra of CBA and D13 cataleptic line mice than in the AKR line resistant to catalepsy. These genes are considered to be the most likely candidate genes of the catalepsy. The coexpression of a large amount of genes in these brain structures in sick animals indicates the existence of a complex gene network that regulates hereditary catalepsy.     

Possible Effects of Glyphosate on Mucorales Abundance in the Rumen of Dairy Cows in Germany

Glyphosate (N-phosphonomethyl glycine) is registered as a herbicide for many food and non-food crops, as well as non-crop areas where total vegetation control is desired. Glyphosate influences the soil mycobiota; however, the possible effect of glyphosate residues in animal feed (soybean, corn, etc.) on animal mycobiota is almost unknown. Accordingly, the present study was initiated to investigate the mycological characteristics of dairy cows in relationship to glyphosate concentrations in urine. A total of 258 dairy cows on 14 dairy farms in Germany were examined. Glyphosate was detected in urine using ELISA. The fungal profile was analyzed in rumen fluid samples using conventional microbiological culture techniques and differentiated by MALDI-TOF mass spectrometry. LPS-binding protein (LBP) and antibodies (IgG1, IgG2, IgA, and IgM) against fungi were determined in blood using ELISA. Different populations of Lichtheimia corymbifera, Lichtheimia ramosa, Mucor, and Rhizopus were detected. L. corymbifera and L. ramosa were significantly more abundant in animals containing high glyphosate (>40 ng/ml) concentrations in urine. There were no significant changes in IgG1 and IgG2 antibodies toward isolated fungi that were related to glyphosate concentration in urine; however, IgA antibodies against L. corymbifera and L. ramosa were significantly lower in the higher glyphosate groups. Moreover, a negative correlation between IgM antibodies against L. corymbifera, L. ramosa, and Rhizopus relative to glyphosate concentration in urine was observed. LBP also was significantly decreased in animals with higher concentrations of glyphosate in their urine. In conclusion, glyphosate appears to modulate the fungal community. The reduction of IgM antibodies and LBP indicates an influence on the innate immune system of animals.     

Antagonistic interactions between Aureobasidium pullulans and Fusarium culmorum,a fungal pathogen of winter wheat

The effect of the biological control agent Aureobasidium pullulans (de Bary) G. Arnaud on the development of Fusarium head blight (FHB) on winter wheat and kernel contamination with fungi of the genera Fusarium, Acremonium, Cladosporium and Penicillium was analyzed in a greenhouse experiment. Scanning electron microscopy was used to evaluate the distribution of A. pullulans cells and aggregates on wheat kernels, infection structures of Fusarium culmorum (W.G. Smith) Sacc and the antagonist-pathogen interactions. Biological control with A. pullulans reduced FHB severity by 21.67 % and improved grain filling by 5.02 %, compared with the control treatment. The survival of A. pullulans was good (to 31 cells per kernel), in particular on the surface and in the crease of kernels, including in pathogen-inoculated wheat plants. A. pullulans cells firmly adhered to F. culmorum hyphae, and damaged them. In most cases, autochthonous communities of filamentous fungi of the genera Acremonium and Penicillium developed at a slower rate after kernel inoculation with the pathogen.     

Outbreeding Depression in Atlantic Salmon Revealed by Hypoxic Stress During Embryonic Development

The fitness consequences of outbreeding in wild populations are extremely variable. Heterosis and outbreeding depression can both be observed but the effect of environmental stressors on the occurrence of these phenomena is still poorly understood. We tested the influence of oxygen stress during embryonic development on consequences of outbreeding in wild populations of Atlantic salmon (Salmo salar). We used a common garden experiment and performed crosses within and between salmon populations to study performances of embryos reared under normal and hypoxic conditions. We detected both heterosis and outbreeding depression depending on traits but irrespective of divergence between parental populations. Nevertheless, outbreeding depression was observed almost exclusively under hypoxic conditions and prevailed over heterosis regarding survival during the whole embryonic development. Notably, the post-hatching survival of all between population crosses was approximately 15 % lower than the survival of within-population crosses under hypoxic conditions. Different hypoxia reaction norms for post-hatching survival, length and time to hatch were also noticed among within and between populations crosses further indicating outbreeding depression. These results demonstrate that consequences of outbreeding can dramatically vary depending on environmental conditions with outbreeding depression being possibly stronger under stressful conditions.