MOSP - Macao Scientific Publishers

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Genetic analysis of Karakul sheep breed using microsatellite markers

Shahram Nanekarani*, Cyrus Amirinia and Nour Amirmozafari

Abstract: In this study, the genetic variation in Karakul sheep was investigated using 15 microsatellite markers (MCMA2, BMS460, BM1815, OARCP26, OARFCB20, MAF64, OARAE129, BMS332, LSCV38, BM6444, BMS995, MCMA26, BMS678 and OARCP49) and all fifteen loci were amplified successfully. Genomic DNA was extracted from 120 blood samples, using modified salting-out method. Tests of genotype frequencies for deviation from the Hardy-Weinberg equilibrium (HWE) were performed at each locus and revealed significant departure from HWE (P < 0.001) due to heterozygote excess. Parameters of variability such as effective number of alleles and gene diversities corroborated with the high level of variation frequently displayed by microsatellite markers. The fifteen tested loci were all polymorphic. Furthermore, other criteria of genetic variation including polymorphism information content (PIC) values and Shanon information index were calculated in this study. Results showed that, high level of genetic diversity was observed in Karakul breed and this breed was not at risk for conservation concept. This research has also shown that microsatellite technique is a useful tool for evaluation of genetic variation among domesticated animals.[...] Read More.

Keywords: Karakul sheep, microsatellite markers, genetic variation, polymorphism. Hardy-Weinberg equilibrium (HWE).

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Antimicrobial activities of the whole plant of Cestrum nocturnum against pathogenic microorganisms

Murad Ali Khan*, Humaira Inayat, Haroon Khan, Mohammmad Saeed, Ikhtair Khan and Inayat-Ur-Rahman

Abstract: The crude methanol extract of the whole plant of Cestrum nocturnum L. (Solanaceae) and its subsequent fractions were tested against various bacterial and fungal strains. With the exception of Salmonella typhi, the tested samples showed marked antibacterial activity against Pseudomonas aeruginosa, Staphylococcus aurous, Bacillus subtilis, Escherichia coli and Shigella flexenari. The minimum inhibitory concentrations (MICs) ranged from 19 to 280 µg/ml. The crude extract and fractions were also susceptible to Candida species and Microsporium canis. The minimum inhibitory concentrations (MICs) for various fungi ranged from 170 to 290 µg/ml. In phytochemical analysis, the crude form and fractions of plant showed the presence of various phytochemical chemical groups like glycosides, alkaloids, saponins, phenols, flavonoids, sterols and tannins. Therefore, the current findings can be attributed to these groups.[...] Read More.

Keywords: Cestrum nocturnum L., antibacterial, antifungal, phytochemicals.

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Preservation methods of fungi in 35 years old stock culture storages: A comparative study

Roseli Santos Freitas*, Kátia Cristina Dantas, Cristiane Neves Pereira, José Eduardo Levi and José Eduardo Costa Martins

Abstract: The maintenance of fungi isolates and the development of methodologies were available from their phenotypic and genotypic characteristics. Alterations of Histoplasma capsulatum and Candida spp. Strains preserved by continuous subculturing on fungal collection and by lyophilization for 35 years were evaluated. Candida isolates maintained by lyophilized methodology were viable and preserved by the typical characteristics of each species. Only one lyophilized isolate from H. capsulatum was viable and demonstrated typical morphology. Both genus, which were preserved by continuous subculturing, revealed morphologic alterations and lost their sporulation capacity. The DNA from these isolates was sequenced (conserved 28S rDNA) in order to confirm their identity. Random amplified polymorphism DNA (RAPD)-based comparative analysis of the two preservation methods revealed alterations in the band profiles in 28 and 33% in Candida spp. and Histoplasma capsulatum strains respectively. The RAPD-based results confirm that the subculturing method alters phenotypic and genotypic characteristics by deleting or inserting nucleotides. Otherwise, the lyophilization was effective to yeasts, but it was not effective for dimorphic fungi.[...] Read More.

Keywords: Polymorphism, genotypic characteristics, lyophilization, random amplified polymorphism DNA.

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Screening of Pogostemon parviflorus Benth. for anti-Candida activity

Shahla Najafi and Batool Sadeghi-Nejad*

Abstract: Pogostemon parviflorus, an aromatic woody herbaceous plant, is found in high rainfall localities. The medicinal properties of this plant show that leaves are used for anxiety, cellulite, depression, eczema and wounds. Also, they are used for thrush, mycotic enteritis and vaginitis. The aim of the present study is to assay the anti-Candida activity of the ethanolic and methanolic extracts of Pogostemon parviflorus leaf against opportunistic mycosis causing phatogenic fungi, such as Candida albicans (5), Candida glabrata (2), Candida tropicalis (2) and Candida dubliensis (1), by agar well diffusion method. Antifungal activity of the ethanolic and methanolic extracts of Pogostemon parviflorus leaf was characterized by ranges of inhibition zones from 8 to 15 mm and 8 to 20 mm, and minimum inhibitory concentration (MIC) by ranges of 2.5 to 20 mg ml−1 and 2.5 to 10 mg ml−1, respectively. The ethanolic extract of tested plant has more anti-Candida effect at 5.7 mg ml−1 when compared to the methanolic extract at 6.6 mg ml−1. The results prove Pogostemon parviflorus leaf as a potent source of natural anti-Candida compounds.[...] Read More.

Keywords: Anti-Candida activity, Pogostemon parviflorus.

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Interaction of Mycobacterium tuberculosis MPB64 protein with heat shock protein 40

Lei Chen*, Xiaowei He, Xihong Zhao and Jianyu Su

Abstract: The MPB64 protein of Mycobacterium tuberculosis (M.tb) is an important structural and functional protein, which has been known to be involved in the virulence, pathogenesis as well as proliferation of the pathogen, however, how MPB64 protein interaction with host protein is still unclear. To identify cellular proteins that interact with the MPB64 protein and to elucidate the possible involvement of MPB64 protein in M.tb pathogenesis, a human lung cDNA library was screened using a yeast two- hybrid system assay. HSP40, a molecular chaperone facilitating protein the folding and assembly, was found to interact specifically with the MPB64 protein. The interaction between MPB64 and HSP40 was verified by colocalization experiment and coimmunoprecipitation of HeLa cell lysates expressing both proteins. The mapping studies localized the critical MPB64 sequences for this interaction to amino acid 171-206. Based on these results, we speculate that HSP40 is a functional target of M. tuberculosis MPB64 protein in cells. This is the first report demonstrating the interaction of HSP40 with a structural protein of M. tuberculosis, indicating a new drug target for M.tb.[...] Read More.

Keywords: MPB64 protein, HSP40 protein, Mycobacterium tuberculosis, yeast two-hybrid, co-immunoprecipitation, protein-protein interaction.

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A simplified mathematical model of multi-species biofilm for simultaneous removal of sulfur dioxide (SO2) and nitric oxide (NO) using a biotrickling-filter

Full Length Research Paper

Abstract: The population dynamics of a multi-species biofilm for simultaneous removal of nitric oxide (NO) and sulfur dioxide (SO2) in a laboratory-scale biotrickling-filter was described. Based on a simplified multispecies-multisubstrate model with the gas-liquid-biomembrane transfer theory, the calculations were compared to the long-term verification experiments. Simulation results were in good agreement with the experimental values of biofilm thickness, population densities of biofilm, total biomass dry weight and partial pressures of NO and SO2 in the outlet gas. The model has been proven to be capable of describing the dynamic biofilm growth, multiple biomass evolution and synergetic effect between sulfate reducing bacteria and denitrifying bacteria on simultaneous removal of NO and SO2. When NO feed concentration was constant at 1780 mg/m3 and SO 2 feed concentration was shifted between 0 and 3200 mg/m3 every half hour, the removal efficiencies of NO and SO2 at steady-state were above 90 and 95%, both in the simulation and experiment.[...] Read More.

Keywords: Population dynamics, multispecies-multisubstrate, gas-liquid-biomembrane transfer, biotrickling-filter, simultaneous SO2 and NO removal.

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In vitro effect of tigecycline against Mycobacterium tuberculosis and a review of the available drugs for tuberculosis

Ahmet Yilmaz Coban*, Aydin Deveci, Yeliz Tanriverdi Cayci, Meltem Uzun, Alper Akgunes and Belma Durupinar

Abstract: The study aimed to investigate the effectiveness of a novel antibiotic drug tigecycline on clinical isolates of Mycobacterium tuberculosis and, reviewing defined anti-tuberculosis effects of available agents. Minimal inhibitory concentration (MIC) of tigecycline for 50 M. tuberculosis including multidrug-resistant (MDR) clinical isolates (20 MDR isolates) was determined by broth microdilution method in the study. Tigecycline MIC values were ranging between 8 and 64 µg/ml. However, there is not any defined break point for M. tuberculosis resistance. In conclusion, it seems that the in vitro effectiveness of tigecycline to M. tuberculosis is not good but further in vivo studies are needed.[...] Read More.

Keywords: Mycobacterium tuberculosis, tigecycline.

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Sensitive and specific detection of Agrobacterium tumefaciens in soil using a rapid polymerase chain reaction (PCR)

Wei Yang, Lei Ji, Li-Rong Tan, Shi-Mo Li, Yan Wang, Hong-Xia Liu*, and Yu-Ming Luo*

Abstract: One pair of primers was designed based on the sequence of tmr locus for specific and sensitive detection of Agrobacterium tumefaciens. Only the A. tumefaciens strain can produce the 236bp target fragment among the fourteen bacterial species that tested. The sensitivity of the specific PCR system was determined by a nested-PCR amplification which can numbered the copies of the template DNA. According to the results, it can give positive band when only 100 copies were in the template. The protocol was carried out for detection A. tumefaciens of twelve soil samples collected from six different gardens in Shanghai where crown gall happened. Two of the samples which collected from symptomless gardens also give the positive band. Based on the results we can make a conclusion that this pair of primers can be a useful tool in detecting A. tumefaciens, especially in detecting latent infection of this devastating pathogen.[...] Read More.

Keywords: Agrobacterium tumefaciens, detection, polymerase chain reaction (PCR).

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Degradation of purine ribonucleosides by extracts of Penicillium viridicatum

Ali M. Elshafei*, Latifa A. Mohamed and Mohamed M. Hassan

Abstract: Cell-free extracts of nitrate-grown Penicillium viridicatum could catalyze the hydrolytic cleavage of N-glycosidic bond of adenosine, guanosine and inosine to the corresponding base and ribose by a ribonucleoside hydrolase; however, there is no evidence for the degradation of these compounds through phosphorylation. The rate of hydrolysis of the three ribonucleosides was in the order inosine> guanosine> adenosine. It was proven that adenosine hydrolyzing enzyme is not associated with the cell membrane. Maximum enzyme activity was observed at pH 4 and 50°C. Heat inactivation kinetics and the effect of the nature of the buffer on enzyme activity revealed that, the cleavage of the three purine ribonucleosides is affected by one intracellular nucleoside hydrolase. It was proven experimentally that, all the metal ions tested had a remarkable inhibitory effect on the activity of the ribonucleoside hydrolase. Results obtained indicate that extracts of P. viridicatum catalyzed the conversion of guanosine into guanine and ribose by a nucleoside hydrolase and the resulting guanine was then deaminated to xanthine by an inducible guanine deaminase. In addition, xanthosine was not split into ribose and xanthine by the same extracts under the same experimental conditions and even at different pH values of the reaction mixture.[...] Read More.

Keywords: Purine ribonucleosides, adenosine, guanosine, inosine, hydrolase, Penicillium viridicatum.

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Anthrax in animals of the Eastern Mediterranean region of Turkey

Ekrem Kirecci*, Suleyman Aslan and Nevin Turut

Abstract: In this study, we aim to perform a microbiological evaluation on the anthrax cases encountered in small and large ruminants as well as in equidae in the Eastern Mediterranean region of Turkey between March 2005 and July 2010. Blood specimens from 44 animals raised in the Mediterranean region of Turkey and suspected of having anthrax in light of the clinical examination results between 2005 and 2010, were subjected to bacteriological analysis. Conventional methods such as Giemsa and Gram staining as well as culture and motility tests were carried out. Smears prepared from blood specimens were treated with Giemsa and Bacillus anthracis pathogens with typical capsules were investigated. During culture test, specimens were cultivated in nutrient broth, blood agar, and MacConkey agar after which they were incubated at 37°C for 24 - 48 h in an aerobic environment. Colony morphologies and microscopic appearances of the bacteria that grew in the nutrient broth and blood agar were evaluated. Thus, bacteria displaying an appearance of typical hair-like shape and a configuration of long chains similar to bamboo sticks, with nonmotile and R-form like colony view, were diagnosed as B. anthracis. The animals with suspected anthrax in this study were comprised of 19 cattle, 14 goats, 10 sheeps and 1 mule. In 12 (27.3%) of 44 specimens, B. anthracis was isolated and identified. B. anthracis was isolated from the animals with suspected anthrax. Anthrax, recognized as a dangerous zoonosis across the entire world, is still of endemic status in our country and preventive measures should include prevention of illegal animal slaughter by performing efficient supervision, applying routine vaccination programs and raising awareness among the livestock owners.[...] Read More.

Keywords: Anthrax, animals, Bacillus anthracis, pathogens.

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