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1.
The objective of this work was to isolate a microorganism producing alkaline protease that can be used as an ecofriendly alternative to chemicals in dehairing process of leather manufacture. Alkaline protease producing bacterium Vibrio metschnikovii NG155 was isolated from soil samples of leather industry. The protease was highly effective in dehairing of goat skin, completely eliminating the use of lime and sulfide. Histological studies of the skin after dehairing showed that the enzyme did not damage the collagen layer and brought good fiber opening. Absence of collagenase activity was confirmed by reacting pure collagen with the enzyme and analyzing it on SDS PAGE, which showed no degradation of collagen. The enzyme was stable in a wide range of pH (7–11) and temperature (10–50 °C), which makes it suitable for industrial application.  相似文献   

2.
Proteases have prospective financial and environment-friendly applications; hence attention is focused currently on the finding of new protease producing microorganism so as to meet the requirements of industry. A thermophilic bacterial strain producing extracellular protease activity was isolated from soil and identified as Bacillus cereus by analysis of 16S rRNA. Protease production by the microorganism was improved by studying the impact of the type of nitrogen and carbon source, fermentation period, growth temperature and initial pH of the culture medium in cultivation optimization experiments. The enzyme was purified to homogeneity in two step procedure involving Sephadex G-75 and Q-Sepharose chromatography. The molecular weight of purified enzyme was found to be 58 kDa by SDS-PAGE. Protease exhibited a pH and temperature optima of 7.5 and 60°, respectively. The enzyme was active in the pH range of 6.0–9.0 and stable up to 70°C. Histological analysis of protease treated goat and cow skin pelts showed complete removal of non leather forming structures such as hair shaft, hair follicles and glandular structures. The protease showed the stain removing property from blood stained cotton cloth and found to be compatible with six commercially available detergents. The protease could release peptides from natural proteins after digestion of coagulated egg albumin and blood clot.  相似文献   

3.
Anti-bacterial drug resistance is one of the most critical concerns among the scientist worldwide. The novel antimicrobial decapeptide SD-8 is designed and its minimal inhibitory concentration and therapeutic index (TI) was found in the range of 1–8 μg/ml and 45–360, respectively, against major group of Gram positive pathogens (GPP). The peptide was also found to be least hemolytic at a concentration of 180 μg/ml, i.e., nearly 77 times higher than its average effective concentration. The kinetics assay showed that the killing time is 120 min for methicillin-sensitive Staphylococcus aureus (MSSA) and 90 min for methicillin-resistant S. aureus (MRSA). Membrane permeabilization is the cause of peptide antimicrobial activity as shown by the transmission electron microscopy studies. The peptide showed the anti-inflammatory property by inhibiting COX-2 with a K D and K i values of 2.36 × 10−9 and 4.8 × 10−8 M, respectively. The peptide was also found to be effective in vivo as derived from histopathological observations in a Staphylococcal skin infection rat model with MRSA as causative organism.  相似文献   

4.
In this study, the methanol extract of Arthrospira (Spirulina) platensis was examined for acute and subchronic toxicities. The extract did not produce any sign of toxicity within 7 days after feeding it at a single high dose of 6 g kg−1 body weight to female and male Swiss mice. For the subchronic toxicity test, the extract at doses of 6, 12, and 24 mg kg−1 body weight was orally administered to six male and six female Wistar rats daily for 12 weeks. Throughout the study period, we did not observe any abnormalities on behavior, food and water intakes, and health status among the treated animals. The hematology and clinical chemistry parameters of treated groups did not significantly differ from those of the controls in both sexes. Postmortem examination of the test groups also showed no abnormalities in both gross and histological findings. These results thus suggest that the methanol extract of A. platensis did not cause acute or subchronic toxicity in our experimental animals.  相似文献   

5.
Bacterial contamination is a serious problem in plant tissue culture procedures. An experiment was conducted to evaluate the potential of nano silver (NS) to remove bacterial contaminants of valerian nodal explants. This experiment was conducted as a completely randomized design in a factorial arrangement with four replications and each replicate with ten explants. Treatments involved NS at two stages (before and after surface sterilization along with control) with three rates (25, 50 and 100 mg l−1) at three times of soaking (30, 60 and 180 min). Explants were cultured on MS medium supplemented with 5 mg l−1 Kin and 0.1 mg l−1 NAA. Results showed that using 100 mg l−1 of NS solution after surface sterilization resulted in the highest percentage (89%) of disinfected explants. Nano silver solution did not affect the characters measured. On the basis of the data obtained in this experiment, it was concluded that NS had a good potential for removing of the bacterial contaminants in plant tissue culture procedures. As this is the first report on application of NS in in vitro culture techniques, further investigations on other plant species are needed to clarify the effectiveness of NS for the removal of bacterial contaminants in tissue culture of other crops.  相似文献   

6.
Ciliate and bacterial densities and their link with eutrophication were studied in fourteen shallow lakes in northwest Spain. Total phosphorus (TP) in these lakes varied between 30 μg l−1 and 925 μg l−1 and chlorophyll a concentration (chla) between 0.5 μg l−1 and 107 μg l−1. Bacterial abundance ranged from 1 × 106 to 14 × 106 cells ml−1, while ciliate abundance ranged from 0.6 cells ml−1 to 229 cells ml−1. Lakes were classified into three trophic types from their TP and chla concentrations. Bacterial abundance was significantly correlated with trophic type, as well as with TP and with chla separately, whereas ciliate abundance was only correlated with chla. No significant relationship could be established between bacterial and ciliate abundance across the trophic gradient. A general pattern was observed in the ratios of bacterial abundance to TP and chla concentrations, of decreasing ratios with increases in the nutrient loading. This pattern was not found for ciliates. The dominant zooplankton group in 13 of the 14 lakes studied was Rotifera, which accounted for a mean of 71% of total zooplankton abundance (41% of zooplankton biomass). The positive correlation between bacteria and ciliates with this group, and the absence of any relationship with Cladocera suggest that top down control by cladocerans was weaker in our lakes than previously shown in northern European shallow lakes. Rotifers could be important predators of bacteria in the high-nutrient lakes of our study. Higher slopes of regressions on bacterial abundance towards the hypertrophic range indicate that top-down control was weaker in our lakes than in northern European shallow lakes.  相似文献   

7.
To better understand the bacterial processes in river sediments, it is necessary to investigate the depth-related bacterial communities in the whole sediment profile. Sediment samples were collected to a depth of 25 cm from the Pearl River. Bacterial abundance, activity, cell-specific respiration rate, and diversity were measured, respectively, by 4′, 6-diamidino-2-phenylindole direct count, 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) staining, electron transport system by CTC reduction, and denaturing gradient gel electrophoresis analysis of 16S rRNA amplification fragments. Results showed that the bacterial metabolism activities decreased with the sediment depth. The total bacterial abundance was highest in the surface sediment with 65.1 × 107 cells g−1, and decreased to 11.1 × 107 cells g−1 below 20 cm in the sample location that suffered from heavy sewage inputs. The active bacteria accounted for 7.50–46.7% of the total bacterial number and decreased with the sediment depth. Electron transport system by the CTC reduction showed that bacterial respiration rate declined from 1.093 μmol CTC-formazan h−1 g−1 in the surface sediment to a half in the bottom sediment, while the cell-specific respiration increased significantly with the depth from 3.56 to 93.75 fmol CTC-formazan cell−1. The bacterial diversity also changed with the depth. Beta-Proteobacteria were the dominant species in the surface sediment, whereas Delta-Proteobacteria were the main species below 10 cm. Results of canonical correspondence analysis (CCA) indicated that the distribution of bacteria was affected by the combined effect of various dissolved inorganic matter, while the respiration rate was independent of the nutrient conditions. The specific bacterial distribution contributed to not only the nutrient cycle but also enhanced pollutant decomposition in sediment of the Pearl River. The results showed that some specific bacterial species had a strong activity in the deeper layers. Therefore, the metabolic functions of the deeper bacterial species should not be neglected.  相似文献   

8.
Preslaughter management procedures that decrease fecal contamination of skins/hides are likely to reduce biological hazards on carcass surfaces during slaughter and processing. This trial was conducted to determine the effects of preslaughter spray-washing on stress responses and skin and carcass bacterial counts in goats. Twenty meat goats were slaughtered in two groups (replicate) on 2 different days (10 goats/replicate). Animals were randomly allotted to treatment (1 min spray-wash) or control (no wash) groups (n = 5/(treatment replicate)). Blood and skin swab samples were collected from control and treated animals before and after the washing treatment. Bacterial counts on the carcasses immediately after dressing were also recorded. Treatment, sampling time or treatment × sampling time did not affect the plasma cortisol, glucose and non-esterified fatty acid (NEFA) concentrations. Skin aerobic plate counts were the same in both treated and control groups prior to washing treatment, but were significantly less in the treated group when sampled after washing (treatment × sampling time, P < 0.05). Aerobic plate counts were 3.6 and 4.4 log10 CFU/cm2 in the treated and control groups, respectively. However, skin Escherichia coli counts did not significantly decrease due to spray-washing treatment. Spray-washing treatment also did not influence carcass E. coli or aerobic plate counts. Results indicate that skin bacterial counts can be significantly reduced by preslaughter spray-washing, without increasing stress in goats. Preslaughter spray-washing may be a cost-effective skin decontamination method that can be easily adopted in goat slaughter plants.  相似文献   

9.
A generic population model is formulated for radiation risk assessment of natural biota. The model demonstrates that effects of radiation on the population survival do not follow directly the effects on individual organisms. Dose rates resulting in population extinction can be analytically calculated. Besides individual radiosensitivity, two key parameters were found to determine the survival potential of a population under chronic radiation stress: the ratio “biomass losses/biomass synthesis,” and the lump amount of limiting resource in the environment. A benchmark scenario “Population response to chronic irradiation” developed within the IAEA Programme EMRAS II was calculated for generic populations of mice, hare/rabbit, wolf/wild dog, and deer/goat chronically exposed to different levels of ionizing radiation. In the conditions of the benchmark scenario, model populations survived normally (>90% of the control value) at dose rates below the following levels: 3 mGy day−1 for wolf/wild dog; 10 mGy day−1 for deer/goat; 14 mGy day−1 for hare/rabbit; and 20 mGy day−1 for mice. The model predictions showed a relatively high survival potential of short-lived and productive species such as mice. At the same time, populations of long-lived animals with slow and radiosensitive reproduction such as wolf/wild dog were candidates to extinction at chronic exposures above 5 mGy day−1. Recovery of short-lived and productive species took a much shorter time compared with long-lived and slow reproductive species.  相似文献   

10.
Photodynamic inactivation is a new promising approach to treat bacterial infections. Usually, the evaluation of the efficacy of this method is done through time-consuming and labor-intensive microbiological test methods. This paper describes the development and implementation of an optical method to evaluate the photodynamic inactivation of bacteria based on non-invasive diffuse reflectance measurements. Five Staphylococcus aureus cultures and 15 mice have been used in this study. A skin lesion was created on the back of all animals, and it was contaminated with S. aureus (5.16 ± 0.013 log CFU/ml). Toluidine Blue O (c = 8.67 × 10 − 3 M) has been used as a photosensitiser agent. The bacterial cultures and animals were exposed to laser radiation (λ = 635 nm, P = 15 mW, DE = 8.654 J/cm2) for 20 min. The photodynamic inactivation of bacteria was monitored by acquiring the wounds’ reflection spectra at different time points and by microbiological exams on the bioptical material. The good correlation between the diffuse reflectance and colony-forming units demonstrates the value of this optical method based on diffuse reflectance measurements as a rapid technique to monitor photodynamic bacterial inactivation.  相似文献   

11.
Studies on the role of endogenous metabolites such as bilirubin and their interactions with biomolecules have attracted considerable attention over the past several years. In this work, the interaction of bilirubin (BR) with purified goat liver cystatin (LC) was studied using fluorescence and ultraviolet (UV) spectroscopy. The fluorescence data proved that the fluorescence quenching of liver cystatin by BR was the result of BR–cystatin complex formation. Stern–Volmer analysis of fluorescence quenching data showed the binding constant to be 9.27 × 104 M−1 and the number of binding sites to be close to unity. The conformation of the BR–cystatin complex was found to change upon varying the pH of the complex. The BR–cystatin complex was found to have reduced papain inhibitory activity. Photo-illumination of BR–cystatin complex causes perturbation in the micro-environment of goat liver cystatin as indicated by red-shift. This report summarizes our research efforts to reveal the mechanism of interaction of bilirubin with liver cystatin.  相似文献   

12.
Endogenous bacterial contaminants isolated from infected cultures of Ilex dumosa nodal segments were identified as Stenotrophomonas maltophilia and Achromobacter sp. using 16S rDNA analysis. A range of antibiotics with different mechanism of actions and the commercial biocide PPM™ were tested for their capacity to repress the growth of Gram negative bacteria grown in liquid medium during the establishment phase of temporal immersion systems. The best results were obtained with the addition of 0.5 mg ml−1 cefotaxime to the culture media obtaining 100% of uncontaminated cultures without suppress of shoot growth.  相似文献   

13.
A thiol proteinase inhibitor from Capra hircus (goat) pancreas (PTPI) isolated by ammonium sulphate precipitation (20–80%) and gel filtration chromatography on Sephacryl S-100HR, with 20.4% yield and 500-fold purification, gave molecular mass of 44 kDa determined by its electrophoretic and gel filtration behavior, respectively. The stokes radius, diffusion and sedimentation coefficients of PTPI were 27.3 Ǻ, 7.87 × 10−7 cm2 s−1 and 3.83 s, respectively. It was stable in pH range 3–10 and up to 70°C (critical temperature, E a = 21 kJ mol−1). Kinetic analysis revealed reversible and competitive mode of inhibition with PTPI showing the highest inhibitory efficiency against papain (K i  = 5.88 nM). The partial amino acid sequence analysis showed that it shared good homology with bovine parotid and skin cystatin C. PTPI possessed 17.18% α helical content assessed by CD spectroscopy. The hydropathy plot of first 24 residues suggested that most amino acids of this stretch might be in the hydrophobic core of the protein.  相似文献   

14.
The alternative transportation strategy implemented during the 2008 Summer Olympic Games in Beijing provided an opportunity to study the impact of the control measures and weather conditions on air quality and asthma morbidity. An ecological study compared the 41 days of the Olympic Games (8 August–17 September 2008) to a baseline period (1–30 June). Also, in order to emphasize the impact of weather conditions on air quality, a pollution linking meteorological index (Plam) was introduced to represent the air pollution meteorological condition. Our study showed that the average number of outpatient visits for asthma was 12.5 per day at baseline and 7.3 per day during the Olympics—a 41.6% overall decrease. Compared with the baseline, the Games were associated with a significant reduction in asthma visits (RR 0.58, 95%CI: 0.52–0.65). At 16.5 visits per day, asthma visits were also significantly higher, during the pre-Olympic period (RR 1.32, 95% CI: 1.15–1.52). The study also showed that the RR of asthma events on a given day, as well as the average daily peak ozone concentration during the preceding 48–72 h, increased at cumulative ozone concentrations of 70 to 100 ppb and 100 ppb or more compared with ozone concentrations of less than 70 ppb (P < 0.05). We concluded that along with “good” weather conditions, efforts to reduce traffic congestion in Beijing during the Olympic Games were associated with a prolonged reduction in air pollution and significantly lower rates of adult asthma events. These data provide support for efforts to reduce air pollution and improve health via reductions in motor vehicle traffic.  相似文献   

15.
Laws of different countries regarding SCC of goat milk are not in agreement with each other and sometimes they fix a threshold for the enhancement of dairy products. The aim of this study was to assess if renneting properties of goat milk are influenced by higher somatic cell count (SCC) measured by an electronic cell counter. Milk samples, taken throughout the lactation of 169 goats from three farms, were analyzed for chemical, physical, hygienic and renneting properties. Samples were divided into three levels on the basis of their SCC: L: low level, samples with SCC lower than 106 cells/ml; M: medium, between 106 and 2 × 106 cells/ml; H: high, higher than 2 × 106 cells/ml. Milk clotting time was between 12.07 and 13.31 min, curd firming time between 1.68 and 2.05 min and curd firmness between 41.66 and 48.97 mm. All the three renneting properties were not affected by the SCC level but they were highly correlated with other factors as protein content and pH. Furthermore, the microbial count showed a high positive correlation with SCC. These results showed that in goat milk, contrarily to other dairy species, higher SCC did not affect renneting properties and that counting of somatic cells by using electronic cell counters might be not suitable for the improvement of dairy products.  相似文献   

16.
Waldrop MP  Firestone MK 《Oecologia》2004,138(2):275-284
Little is known about how the structure of microbial communities impacts carbon cycling or how soil microbial community composition mediates plant effects on C-decomposition processes. We examined the degradation of four 13C-labeled compounds (starch, xylose, vanillin, and pine litter), quantified rates of associated enzyme activities, and identified microbial groups utilizing the 13C-labeled substrates in soils under oaks and in adjacent open grasslands. By quantifying increases in non-13C-labeled carbon in microbial biomarkers, we were also able to identify functional groups responsible for the metabolism of indigenous soil organic matter. Although microbial community composition differed between oak and grassland soils, the microbial groups responsible for starch, xylose, and vanillin degradation, as defined by 13C-PLFA, did not differ significantly between oak and grassland soils. Microbial groups responsible for pine litter and SOM-C degradation did differ between the two soils. Enhanced degradation of SOM resulting from substrate addition (priming) was greater in grassland soils, particularly in response to pine litter addition; under these conditions, fungal and Gram + biomarkers showed more incorporation of SOM-C than did Gram – biomarkers. In contrast, the oak soil microbial community primarily incorporated C from the added substrates. More 13C (from both simple and recalcitrant sources) was incorporated into the Gram – biomarkers than Gram + biomarkers despite the fact that the Gram + group generally comprised a greater portion of the bacterial biomass than did markers for the Gram – group. These experiments begin to identify components of the soil microbial community responsible for decomposition of different types of C-substrates. The results demonstrate that the presence of distinctly different plant communities did not alter the microbial community profile responsible for decomposition of relatively labile C-substrates but did alter the profiles of microbial communities responsible for decomposition of the more recalcitrant substrates, pine litter and indigenous soil organic matter.  相似文献   

17.
Transmembrane Bax Inhibitor Motif-containing 6 (TMBIM6) gene acts as calcium leak channel and negatively regulates autophagy and autophagosome formation. The TMBIM6 gene was amplified and searched for variation in three different goat populations (i.e. Black Bengal, Ganjam and Raighar) of Odisha state of the India. The result indicated two substitutions i.e. 55th position (C55T) and 95th position (C95A) in the amplified region of the gene resulting in change of amino acids (Leu > Phe and Thr > Asn). The identified SNPs were combined to form haplotypes and animals were grouped accordingly. Structural analysis showed minor changes (5%) in between mutant and wild TMBIM6 protein structures. However, any functional variation could not be identified with respect to the calcium ligand and open pore state. But an alteration of calcium binding site was found. The binding interaction of calcium with the TMBIM6 protein was hydrophobic in nature in closed state whereas hydrophilic in open pore stage. The stress releasing function was the result of calcium leakage controlled by amino acids coded by exon 4 and exon 5 regions of TMBIM6 gene. The effect of breed and haplotype on cardiopulmonary traits was studied. The data on cardiopulmonary traits of body i.e. rectal temperature, skin temperature, heart rate and respiration rate were recorded when ambient temperature usually remained the highest. The statistical analysis showed, significant difference in rectal temperature, skin temperature and respiration rate among these goat populations. The haplotypes (CC and TA) were found to have a significant (P < 0.05) effect on rectal temperature, skin temperature and respiration rate. However, any such significant effect could not be identified in recorded heart rate. The objective of the present study to identify the genetic variations in TMBIM6 gene having significant effect on cardiopulmonary traits which can be further uses as the molecular markers to improve heat tolerance mechanism in goats.  相似文献   

18.
The high polysaccharide content of some plant species hinders the successful isolation of their DNA. As an alternative to the macro-extraction methods previously published for polysaccharide-rich plants, we present two techniques (STE/CTAB and HEPES/CTAB), which are performed in microcentrifuge tubes. These protocols are suitable for small amounts of silica gel-preserved plant tissue such as are commonly available from endangered plants. The critical step to remove polysaccharides was performing initial washes in either STE (0.25 M sucrose, 0.03 M Tris, 0.05 M EDTA) or HEPES (2% β-mercaptoethanol, 0.2% PVP, 0.1 M HEPES, pH 8.0) buffer. Precipitating the DNA at room temperature with isopropanol also aided in decreasing polysaccharide co-precipitation. Of the two protocols we present the STE/CTAB method has the advantages of being more cost-effective and avoiding the use of the hazardous chemical β-mercaptoethanol.  相似文献   

19.
We cloned the cDNA and genomic DNA encoding for Izumo1 of cashmere goat (Capra hircus) and sheep (Ovis aries). Analysis of 4.6 kb Izumo1 genomic sequences in sheep and goat revealed a canonical open reading frame (ORF) of 963 bp spliced by eight exons. Sheep and goat Izumo1 genes share >99% identity at both DNA and protein levels and are also highly homologous to the orthologues in cattle, mouse, rat and human. Extensive cloning and analysis of Izumo1 cDNA revealed three (del 69, del 182 and del 217) and two (del 69 and ins 30) alternative splicing isoforms in goat and sheep, respectively. All of the isoforms are derived from splicing at typical GT-AG sites leading to partial or complete truncation of the immunoglobulin (Ig)-like domain. Bioinformatics analysis showed that caprine and ovine Izumo1 proteins share similar structure with their murine orthologue. There are a signal peptide at the N-terminus (1–22 aa), a transmembrane domain at the C-terminus (302–319 aa), and an extracellular Ig-like region in the middle (161–252 aa) with a putative N-linked glycosylation site (N205-N-S). Alignment of Izumo1 protein sequences among 15 mammalian species displayed several highly conserved regions, including LDC and YRC motifs with cysteine residues for potential disulfide bridge formation, CPNKCG motif upstream of the Ig-like domain, GLTDYSFYRVW motif upstream of the putative N-linked glycosylation site, and a number of scattered cysteine residues. These distinctive features are very informative to pinpoint the important gene motifs and functions. The C-terminal regions, however, are more variable across species. Izumo1 cDNA sequences of goat, sheep, and cow were found to be largely homologous, and the molecular phylogenetic analysis is consistent with their morphological taxonomy. This implies the Izumo1 gene evolves from the same ancestor, and the mechanism of sperm–egg fusion in mammals may be under the same principle in which Izumo1 plays an important role.  相似文献   

20.
Tributyl phosphate (TBP) is an organophosphorous compound, used extensively (3000–5000 tonnes/annum) as a solvent for nuclear fuel processing and as a base stock in the formulation of fire-resistant aircraft hydraulic fluids and other applications. Because of its wide applications and relative stability in the natural environment TBP poses the problem of pollution and health hazards. In the present study, fifteen potent bacterial strains capable of using tributyl phosphate (TBP) as sole carbon and phosphorus source were isolated from enrichment cultures. These isolates were identified on the basis of biochemical and morphological characteristics and 16S rRNA gene sequence analysis. Phylogenetic analysis of 16S rRNA gene sequences revealed that two isolates belonged to class Bacilli and thirteen to β and γ-Proteobacteria. All these isolates were found to be members of genera Alcaligenes, Providencia, Delftia, Ralstonia, and Bacillus. These isolates were able to tolerate and degrade up to 5 mM TBP, the highest concentration reported to date. The GC–MS method was developed to monitor TBP degradation. Two strains, Providencia sp. BGW4 and Delftia sp. BGW1 showed respectively, 61.0 ± 2.8% and 57.0 ± 2.0% TBP degradation within 4 days. The degradation rate constants, calculated by first order kinetic model were between 0.0024 and 0.0099 h−1. These bacterial strains are novel for TBP degradation and could be used as an important bioresource for efficient decontamination of TBP polluted waste streams.  相似文献   

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