A convenient and sensitive fluorescence assay for phospholipid vesicles using diphenylhexatriene

When phospholipid vesicles are added to an aqueous solution of 1,6-diphenyl-1,3,5-hexatriene (DPH) a fluorescence enhancement of up to several hundredfold is observed which can be used for a determination of phospholipid concentration. Fluorescence enhancement of 2 μm DPH is proportional to the phospholipid concentration over a wide range. As little as 0.7 nmol (～0.5 μg of phospholipid) can be determined to within ±10%. The fluorescence is a function of the type of phospholipid used, salt concentration, and time of incubation. Protein and detergents also enhance DPH fluorescence but to a much smaller extent. Optimal conditions for the assay are presented. Use of this assay to detect phospholipid vesicles fractionated by size on a Sepharose 4B column is illustrated. In this case the method compares favorably to more classical methods of analysis in terms of sensitivity, accuracy, and time involved.     

Physicochemical and structural characterization of biosurfactant produced by <Emphasis Type="Italic">Pleurotus djamor</Emphasis> in solid-state fermentation

Biosurfactants are amphiphilic compounds produced by several microorganisms that reduce the surface tension. Low toxicity, optimal activity in extreme conditions, biodegradability and production from several wastes are main advantages of biosurfactants as compared to synthetic surfactants. Production of biosurfactant by a white rot fungus Pleurotus djamor on sunflower seed shell in solid-state fermentation was determined by emulsification indexes, oil spreading activity and surface tension (28.82 ± 0.3mN/m) measurement. The critical micelle concentration was detected as 0.964 ± 0.09 mg/mL. Also, the chemical and physicochemical properties of the biosurfactant produced were investigated. Considering the results of the chemical contents analysis, HPLC, FT-IR and ¹H-NMR, it can be concluded that the produced biosurfactant has a complex structure. Besides, resistance of its activity to environmental factors such as temperature, pH and salt concentration, as well as its thermal stability, were investigated. Additionally, the produced biosurfactant formed stabile emulsions with different hydrocarbons. Lastly, the performance of removing waste frying oil from contaminated sand of produced biosurfactant was detected as 76.57 ± 6%. Owing to its high emulsification capacity, low surface tension and critical micelle concentration, the biosurfactant, shows great potential for use in hydrocarbon removal applications.     

不同浓度胆汁酸盐对人肝胆管癌细胞凋亡的影响

目的:探讨不同浓度胆汁酸盐对人肝胆管癌细胞RBE凋亡的影响。方法:采用0、100μM、500μM、1 m M胆汁酸盐作用于人肝胆管癌细胞后,采用流式细胞术检测细胞的凋亡率,Western-blot法检测细胞Bax、Bcl-2蛋白的表达。结果:胆汁酸盐浓度为0、100μM、500μM、1 m M时,人胆管癌细胞的凋亡率分别为(0.7±0.12)%、(0.9±0.15)%、(1.4±0.17)%、(4.8±0.14)%,以1 m M胆汁酸盐所致人胆管癌细胞的凋亡最明显,Bcl-2蛋白表达量逐渐减少,Bax表达量逐渐增多,以1 m M胆汁酸盐时作用最明显,差异具有统计学意义(P0.05)。1 m M胆汁酸盐处理的人肝胆管癌细胞的形态由原来的梭形变成圆形,细胞核固缩、碎裂。结论:胆汁酸盐可以以浓度依赖性的方式导致人胆管癌细胞凋亡,以1 m M胆汁酸盐的作用最明显。     

Reference values for exhaled nitric oxide (reveno) study

Background

Human airway surface liquid (ASL) has abundant antimicrobial peptides whose potency increases as the salt concentration decreases. Xylitol is a 5-carbon sugar that has the ability to lower ASL salt concentration, potentially enhancing innate immunity. Xylitol was detected for 8 hours in the ASL after application in airway epithelium in vitro. We tested the airway retention time of aerosolized iso-osmotic xylitol in healthy volunteers.

Methods

After a screening spirometry, volunteers received 10 ml of nebulized 5% xylitol. Bronchoscopy was done at 20 minutes (n = 6), 90 minutes (n = 6), and 3 hours (n = 5) after nebulization and ASL was collected using microsampling probes, followed by bronchoalveolar lavage (BAL). Xylitol concentration was measured by nuclear magnetic resonance spectroscopy and corrected for dilution using urea concentration.

Results

All subjects tolerated nebulization and bronchoscopy well. Mean ASL volume recovered from the probes was 49 ± 23 μl. The mean ASL xylitol concentration at 20, 90, and 180 minutes was 1.6 ± 1.9 μg/μl, 0.6 ± 0.6 μg/μl, and 0.1 ± 0.1 μg/μl, respectively. Corresponding BAL concentration corrected for dilution was consistently lower at all time points. The terminal half-life of aerosolized xylitol obtained by the probes was 45 minutes with a mean residence time of 65 minutes in ASL. Corresponding BAL values were 36 and 50 minutes, respectively.

Conclusion

After a single dose nebulization, xylitol was detected in ASL for 3 hours, which was shorter than our in vitro measurement. The microsampling probe performed superior to BAL when sampling bronchial ASL.     

Selection and Combustion of Ni-Hyperaccumulators for the Phytomining Process

Ni recovery from serpentine soils by phytomining has proved feasible. Phytomining involves the crop of hyperaccumulating plants with high Ni contents and the valorization of Ni by pyro or hydrometallurgical process. In order to evaluate the Ni content of different plants, we analyzed the organs of 14 hyperaccumulators from three genera: Alyssum, Leptoplax and Bornmuellera. The highest concentration was recorded in the leaves of Leptoplax (34.3 ± 0.7 mg g^?1 _DM). Additionally, we investigated biomass combustion which is the first step of the process we designed to obtain a nickel salt. We showed that temperature and duration were important parameters to ensure a good quality of ashes. At the bench scale, the best conditions were 550°C and 3 h. In this way, we obtained ashes in which Ni could reach 20 wt%. Biomass ashes can be considered as a bio-ore for recovering metal value.     

Influence of polymerization method on the cytotoxicity of three different denture base acrylic resins polymerized in different methods

Statement of ProblemAcrylic plastics are used for over 80 years for the manufacture of prostheses. This kind of material has some limitations, one of them is a residual monomer, that remains after the polymerization has been terminated, which can influence the biological properties of the final medical device.The purposeThe aim of this investigations was a comparison of the residual monomer concentration and cytotoxic effect of three various acrylic materials which differ in the polymerization method (hot-cured, polymerized under pressure and at lower temperatures).Material and methodsThe cytotoxicity of three different acrylic resins from the same producer were tested on the in vitro model (VERO CCL-81) by MTT assay. The residual monomer of acrylic materials was detected by gas chromatography.ResultsThe Superacryl Plus material polymerized in hot water has a residual monomer concentration of 0.67 ± 0.05%, Superpont C + B hardened under pressure of 2.61 ± 0.208%, and Premacryl Plus after cold curing process has 3.53 ± 0.27% of uncured MMA. The results revealed that the least cytotoxic effect were observed in case of a thermally polymerized material.ConclusionMaterial polymerized in high temperatures has lower residual monomer concentration and not affect cell cultures. Self-curing materials polymerized in lower temperature have a higher concentration of residual monomer, which reduces the number of living cells by 20%, which can cause allergic reaction shortly after new denture was prepared.     

A convenient pH-gradient method for the determination of the maximum and minimum pH for microbial growth

Previously existing methods for determining the pH limits for the growth of microorganisms have involved (1), the setting up of individual cultures, each having a specific pH; (2), the pH gradient plate technique devised by Sacks (1956) in which a continuous pH gradient is established in a Petri dish by means of a buffer system; and (3), the pH gradient plate technique of Zak (unpublished), in which a continuous pH gradient is established by means of an electric current. The discontinuous pH gradient technique described here provides a convenient method of determining the maximum and minimum pH at which a microorganism can grow. The technique can be used aerobically or anaerobically, and has a precision of about ± 0.1 pH unit. Data are given for several yeasts and forSerratia marcescens. In all cases, the organisms tested continued to metabolize at pH values beyond those representing the limits for growth, sometimes by as much as 0.5 pH unit. The results suggest that pH limits are unsuitable criteria in microbial classification.     

高温湿热酸法破壁提取法夫酵母胞内虾青素

法夫酵母是一种能积累虾青素的红酵母, 对其进行破壁是当前虾青素工业化提取生产的瓶颈工艺。研究在高温湿热条件下,低浓度盐酸对法夫酵母破壁而提取其胞内虾青素的工艺。探讨了不同破壁温度、盐酸浓度、液料比与破壁处理时间等因素对法夫酵母破壁后虾青素及类胡萝卜素提取率的影响, 确定了高温湿热酸法破壁提取虾青素的最佳条件为: 饱和蒸汽压力 0.1 MPa (121°C), 盐酸浓度0.5 mol/L, 液料比 (V/W)30 mL/g, 破壁时间2 min。在最佳条件下进行中试放大实验, 可得到虾青素与类胡萝卜素的提取率分别为: (84.8±3.2)%, (93.3±2)%。经优化后的新破壁工艺安全高效, 不会有毒性残留, 具有良好的工业应用前景。     

A study of the efficiency of edible oils degraded in alkaline conditions by <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> SS-219 and <Emphasis Type="Italic">Acinetobacter</Emphasis> sp. SS-192 bacteria isolated from Japanese soil

High lipid concentration contained in wastewater inhibits the activity of microorganisms in biological wastewater treatment systems such as activated sludge and methane fermentation. To reduce the inhibitory effects, microorganisms capable of efficiently degrading edible oils were screened from various environmental sources. From Japanese soil, we isolated 2 bacteria strains with high degradation abilities at an alkaline pH without consumption of biological oxygen demand (BOD) constituents. Acinetobacter sp. strain SS-192 and Pseudomonas aeruginosa strain SS-219 degraded 77.5 ± 0.6% and 89.5 ± 1.5%, respectively, of 3,000 ppm of mixed oil consisting of salad oil/lard/beef tallow (1/1/1, w/w/w) at 37°C and pH 9.0 in 24 h. Efficient degradation by the two strains occurred at pH 8–9 and 25–40°C. Strain SS-219 degraded lipids even at pH 3. The degradation rate of 3,000 ppm of salad oil, lard, and beef tallow by strain SS-192 was 79.9 ± 2.6%, 63.6 ± 1.9%, and 70.1 ± 1.2%, respectively, during a 24-h cultivation. The degradation rate of 3,000 ppm of salad oil, lard, and beef tallow by strain SS-219 was 82.3 ± 2.1%, 71.9 ± 2.2%, and 71.0 ± 1.1%, respectively, during a 24-h cultivation. After mixed oil degradation by both strains, the BOD value of the cell culture increased from 2,100 ppm to 3,200–4,000 ppm. The fact that neither strain utilizes BOD ingredients will be beneficial to pretreatment of methane fermentation systems such as upflow anaerobic sludge blanket reactors. In addition, the growth of usual heterotrophic microorganisms utilizing soluble BOD can be suppressed under alkaline pH.     

Optimization of Operating Parameters for the Selective Flotation of Heavy Metals from Contaminated Fine Sediment Using Response Surface Model

In this study, the effect of different flotation operating variables, such as pH, pulp density, collector concentration, impeller speed, frother concentration, and air flow rate, on selective flotation of heavy metals, especially Cu, from fine dredged sediment has been evaluated. Parameter optimization was done using the single parameter at a time method and response surface method (RSM) using Box-Behnken design and was assessed in terms of metal removal, metal recovery, metal concentration factor, and mass recovery. Among the operating variables studied, pulp pH, collector concentration, pulp density, and impeller speed were found to have significant effect on metal flotation selectivity. A validation study of the response surface model showed its aptness to predict the optimum values of operating parameters and their interactions on flotation responses which evaluate flotation performance. Flotation experiments under optimum operating parameters showed good flotation selectivity for Cu (3.3 ± 0.2) with a mass recovery of (mass of sediment in the froth) 14.1 ± 1 and Cu removal of 37.4 ± 3.6%.     

Distribution and fate of anthropogenic nitrogen in the Calamagrostis angustifolia wetland ecosystem of Sanjiang Plain, Northeast China

Wetlands are important for the protection of water quality of rivers and lakes, especially those adjacent to agricultural landscapes, by intercepting and removing nutrients in runoff. In this study, the 15N tracer technique was applied to study the distribution and fate of anthropogenic nitrogen (15N-fertilizer) in Calamagrostis angustifolia Kom wetland plant-soil microcosms to identify the main ecological effects of it. 15NH415NO3 solution (14.93 mg N/L, 20.28 at.% 15N) was added to each microcosm of the first group, which was approximate to the current nitrogen concentration (CNC) of farm drainage, and 29.86 mg NIL 15NH415NO3 solution was added to another group, which was approximate to the double nitrogen concentration (DNC) of farm drainage, while no nitrogen (NN) was added to the third group. The results suggest that the Input of anthropogenic nitrogen has positive effects on the biomass and total nitrogen content of plant, and the positive effects will be elevated as the increase of its Input amount. The increase of 15N-fartilizer can also elevate its amounts and proportions in plant nitrogen. Soil nitrogen is still the main source of plant nitrogen, but its proportion will be reduced as the increase of 15N-fertilizer. The study of the fate of 15N-fartilizar indicates that, in CNC treatment, only a small proportion is water-dissolved (0.13±0.20%), a considerable proportion is soil-immobilized (17.02±8.62%), or plant-assimilated (23.70±0.92%), and most is lost by gaseous forms (59.15±8.35%). While in DNC treatment, about 0.09±0.15% is water-dissolved, 15.33±7.46% is soil-immobilized, 23.55±2.86% is plant-assimilated, and 61.01±5.59% is lost by gaseous forms. The double input of anthropogenic nitrogen can not elevate the proportions of plant-assimilation, soil-immobilization and water-dissolution,but it can enhance the gaseous losses.     

Bioethanol from Macroalgal Biomass: Utilization of Marine Yeast for Production of the Same

The present study deals with the first systematic study on the isolation, characterization, and utilization of marine yeast for bioethanol production using seaweed biomass. The ability and efficiency of isolated marine yeast to grow and ferment sugar to ethanol in the presence of 2.5 % to 15 % salt concentration was validated by fermenting galactose in the presence of different salts at varied concentrations. Successively, this yeast was employed for fermentation of seaweed hydrolysate, containing high salt concentration, to ethanol. The hydrolysate having varying sugar as well as salt contents, from 2.7 % to 5.5 % and from 6.25 to 11.25 %, respectively, yielded 1.23–1.76 % ethanol. Through biochemical, fatty acid methyl ester analysis, and BioLog, the yeast was identified as Candida sp. The ability of this yeast to function at high salinity can be commercialized for its use to convert seaweed polysaccharide based hydrolysate, rich in salt, to ethanol without desalting process, ultimately making the process more efficient and economically viable. This is the first organized study for the utilization of marine yeast for converting Kappaphycus alvarezii, a red algal biomass, into ethanol as a byproduct, under highly saline condition.     

Antagonistic Interactions between Stress Factors during the Growth of Microorganisms under Conditions Simulating the Parameters of Their Natural Ecotopes

Two stress factors, hypoxia (microaerobic conditions) and a high salt concentration, if applied simultaneously to aerobic microorganisms, display an antagonistic mode of interaction. As a result, the NaCl level that is usually optimal for moderate halophiles (5–6 %) becomes optimal for the growth of weak halophiles (Rhodococcus erythropolis and Shewanella sp. CN32); the halotolerant yeast Yarrowia lypolytica acquires halophilic properties (with a growth optimum at a NaCl concentration of 10%), and the growth rate of the extremely halophilic Halobacterium salinarum increases at supraoptimal salt concentrations (25–34%). This phenomenon is apparently due to multiple changes in metabolic reactions. In particular, high salt concentrations suppress respiration and the formation of enzymes (superoxide dismutase and catalase) that protect the cell from toxic oxygen species. Therefore, establishment of microaerobic conditions compensates for the loss of these protective mechanisms and enables cell growth at higher salt concentrations than under aerobic conditions. Of some importance can also be the increase in the intracellular concentrations of osmoprotectants caused by the suppression of their intracellular oxidation. The implications of this phenomenon for the ecophysiology of microorganisms (including oil-oxidizing species) and for the classification of weak and moderate halophiles are discussed.     

Effect of ursodeoxycholate on the bile flow in the rat

The relationship between the bile flow and biliary excretion rate of bile salt was studied by a continuous infusion of ursodeoxycholate and its glycine conjugate in rats. Infusion of glycoursodeoxycholate produced a higher flow rate and higher bile salt concentration than previously reported values for taurocholate. The estimated biliary transport maximum value was 2.21±0.15 μmole/min/100g body weight (mean±SD, N=13). Furthermore, a linear relation was found between the bile flow and bile salt excretion rate for a wide range of bile salt excretion with a slope value of 4.10±0.64 μl/μmole (N=10). These values were close to values previously reported for tauroursodeoxycholate. In contrast, when free ursodeoxycholate was infused, a bile salt excretion rate increased at first to a level of around 1.0 μmole/min/100g body weight with a concomitant bile flow increase, but after one hr, the bile salt excretion dropped sharply and a lower plateau of about half of the initial maximum level was established in the following hr. On the other hand, the bile flow further increased even in the second hr. Consequently, the linear relationship initially observed between the bile flow and bile salt excretion rate became gradually distorted and after one hr even the positive correlation between the two parameters was completely lost. The sharp drop in the bile salt excretion rate was found to be due to the decrease in the taurine conjugate of ursodeoxycholate in the bile. The excretion rate of free ursodeoxycholate remained at a very low level (about 0.1 μmole/min/100g body weight) throughout the experiments. The concentration of ursodeoxycholate in the liver increased sharply in the second hr corresponding to the decrease in the bile salt excretion rate. These results appear to be most easily explained by the thesis that there is a fraction of bile independent of bile salt excretion but dependent on the bile salt concentration in the hepatocyte.     

Antagonistic interactions between stress factors during the growth of microorganisms under conditions simulating the parameters of their natural ecotopes

Two stress factors, hypoxia (microaerobic conditions) and a high salt concentration, if applied simultaneously to aerobic microorganisms, display an antagonistic mode of interaction. As a result, the NaCl level that is usually optimal for moderate halophiles (5-6%) becomes optimal for the growth of weak halophiles (Rhodococcus erythropolis and Shewanella sp. CN32); the halotolerant yeast Yarrowia lypolytica acquires halophilic properties (with a growth optimum at a NaCl concentration of 10%), and the growth rate of the extremely halophilic Halobacterium salinarum increases at supraoptimal salt concentrations (25-34%). This phenomenon is apparently due to multiple changes in metabolic reactions. In particular, high salt concentrations suppress respiration and the formation of enzymes (superoxide dismutase and catalase) that protect the cell from toxic oxygen species. Therefore, establishment of microaerobic conditions compensates for the loss of these protective mechanisms and enables cell growth at higher salt concentrations than under aerobic conditions. Of some importance can also be the increase in the intracellular concentrations of osmoprotectants caused by the suppression of their intracellular oxidation. The implications of this phenomenon for the ecophysiology of microorganisms (including oiloxidizing species) and for the classification of weak and moderate halophiles are discussed.     

超滤法和盐析法粗提基因工程干扰素其比活的比较

我们的工作是应用ＹＦ２ＰＳＫ３１Ｒ型超滤器超滤粗提基因工程干扰素,并与盐析法对比。超滤法简便快速条件温和,超滤后干扰素平均收率为９３．９５％±２．４８％,盐析法则为７５．９３％±３．９％,前者比后者提高１８．０２％,差异显著。比活前为：４．４５５Ｅ６±７．５８Ｅ５ＩＵ／ｍｇ蛋白,后者为：１．６１１７Ｅ６±２．０７９Ｅ５ＩＵ／ｍｇ蛋白,差异也是显著的     

Influence of Celery on the Remediation of PAHs-contaminated Farm Soil

Polycyclic aromatic hydrocarbons (PAHs) contamination has been considered as one of the major environmental concerns for farmland soil all over the world including China. Due to small per capita land area, to find crops or vegetable, which could not only degrade the PAHs contaminants but also would not concentrate PAHs, was particularly important. Celery was selected as the phytoremediator in this experiment, and the soil enzyme activity, PAHs-degrading microorganisms, and the speciation of PAHs in soil were studied. The results showed that celery could significantly enhance the remediation of PAHs compared with the controlled experiment after 90 days (p< 0.01), and the removal efficiency were 31.29%, 30.79%, and 50.21% in the soil, non-rhizosphere soil, and rhizosphere soil, respectively. The soil enzyme activity and PAHs-degrading microorganisms significantly increased in rhizosphere soil compared with non-rhizosphere soil (p< 0.05), and the bioaccessibility of PAHs in soil could have been enhanced in the presence of celery root exudates. Those would help the bioremediation of PAHs by soil microorganisms. Meanwhile, the concentration of PAHs in the edible portion of celery was only 17.13 ± 1.24 μg/kg, and the bioconcentration factors in the aboveground part of celery were only 0.025. This study provides a potential in-site farmland soil phytoremediation technology that could have practical utility.     

Plant size metrics and organic carbon content of Florida salt marsh vegetation

Carbon stored in soil and vegetative biomass in mangrove swamps, salt marshes, and seagrass flats (“coastal blue carbon”) is a significant component of the coastal carbon budget due to the ability of these ecosystems to sequester atmospheric carbon dioxide as plant biomass and peat. Growing interest in the protection of these essential yet vulnerable coastal ecosystems, as well as recognition of their capacity to sequester atmospheric carbon dioxide, has led to the need for rapid and accurate quantification of location-specific coastal blue carbon stocks. This study provides allometric equations for the calculation of aboveground dry biomass based on plant height for 18 salt marsh species from Tampa Bay, Florida, USA. The 18 plant species were also analyzed for carbon and nitrogen content to determine the appropriate carbon conversion factor to use for blue carbon calculations. The salt marsh plants had an average of 1.2 ± 0.4% nitrogen and 41.1 ± 5.5% carbon. With the exception of Batis maritima, which had a carbon content of only 23.4%, carbon content for all other species ranged from 33.4–47.1%. The four most succulent plant species in this study contained an average of 33.3 ± 7.0% carbon, while the nine graminoid species contained 44.4 ± 2.0% carbon. These species-specific equations and carbon data provide non-destructive methods to rapidly quantify vegetative biomass and carbon stocks for monitoring efforts and blue carbon stock assessments.     

Salt-induced hypertension in chronic renal failure: Evidence for a neurogenic mechanism

We investigated the possibility that blood pressure elevation induced by salt excess may be secondary to a neurogenic mechanism. The compound SK&F 64139 (50 mg/kg) known to inhibit central and peripheral phenylethanolamine N-methytransferase (PNMT) the enzyme necessary for the conversion of norepinephrine to epinephrine, was given by oral gavage to two groups of subtotally nephrectomized rats maintained for five days on either a high salt (HS) or low salt (LS) diet respectively. Blood urea nitrogen (BUN) and hematocrit were not different between the two groups, while body weight and serum Na were significantly higher in the HS animals. Baseline mean blood pressure (BP) was higher in the HS animals (HS 154 ± 4.7 vs LS 121 ± 3.7 mmHg, p<0.001) and decreased by 39 ± 6.9 mmHg one and one half hour post SK&F 64139 to normotensive levels in the HS as opposed to a decrease of 10 ± 1.8 mmHg in the LS group. Baseline heart rate (HR) was higher in the LS group (474 ± 17 beats/min) vs the HS group (408 ± 17, p<0.05), and decreased significantly after SK&F 64139 in both groups to the same extent (by 17.6% in the HS vs 13.3% in the LS). A third group of subtotally nephrectomized rats maintained for five days on a HS diet were given by oral gavage the compount SK&F 29661 (100 mg/kg), a PNMT inhibitor which does not cross the blood-brain barrier. Following SK&F 29661, there was no significant decrease in mean BP (153 ± 5 to 149 ± 4 mmHg) and a less than 2% decrease in HR. Baseline plasma norepinephrine (NE) was higher in the HS as compared to the LS group (1.50 ± 0.16 vs 0.904±0.15 ng/ml, respectively, p<0.05) and a significant correlation was found between plasma NE level and decrease in BP following SK&F 64139 (r=0.65, p<0.01). Not withstanding possible effects of some ancillary properties of SK&F 64139, these data support the hypothesis that a neurogenic component, possibly mediated via central epinephrine containing neurons, contributes to the BP elevation induced by salt excess.     

Development of a RNA extraction method from milk for gene expression study in the mammary gland of sheep

The aim of the study was to develop a reliable method for the RNA extraction from milk of Sarda sheep breed and to highlight if the extracted RNA can be used for expression study on mammary genes involved in milk fat synthesis using RT-qPCR. The main result is that a sample of 150 ml of milk provides an optimal amount of RNA (73.5 μg/ml). The highest RNA concentration has been found in the samples analysed within 4 h after collection. The RNA extracted was positively correlated to the number of somatic cells (P < 0.001). The efficiency of the extraction method was confirmed by the results obtained from qPCR which showed a Ct value, for SREBPF1 gene of 26.8 ± 0.15. This research demonstrated that the high-quality of the RNA obtained is suited to use for studies of mammary genes expression in sheep, avoiding any damage caused by mammary gland biopsy.