Development of a method for the quantitative assessment of microorganism sensitivity to antibiotics using discs. The study of different nutrient media for determining microorganism sensitivity to antibiotics

Five nutrient media used for determination of microbial sensitivity to antibiotics, i.e. beaf-peptone agar, Hottinger pancreatic beaf infusion agar, sprat hydrolysate nutrient agar of the Dagestan Research Institute of Nutrient Media, Muller-Chintone agar from Bulgaria and "Oxoid" agar for determination of microbial sensitivity were studied comparatively. The media were compared with respect to the growth density with the use of different test-cultures and the clearance of the inhibition growth zones around the discs containing different antibiotics. The best results were obtained with the use of sprat hydrolysate nutrient agar. Further studies on the medium standardization are necessary.     

Determination of the sensitivity of bacteria to barium ions,a taxonomic marker of the genus <Emphasis Type="Italic">Pseudomonas</Emphasis>

Comparative efficacy of the determination of the sensitivity of bacterial cells to barium ions was evaluated on a synthetic nutrient medium, FMH agar, Mueller-Hinton agar, and AGV agar. The synthetic nutrient medium developed for this study contained L-proline and L-glutamine as the sole nitrogen and carbon source, which promoted growth of all Pseudomonas strains and ensured the minimal level of barium binding. The sensitivity of 80 strains belonging to 11 Pseudomonas species, including the type strains, as well as of 80 strains of 22 other bacterial species, was studied. The sensitivity of bacteria to barium ions was determined by using serial dilutions of barium chloride in the nutrient medium. The highest level of analytical sensitivity of pseudomonads to barium ions was determined on the synthetic nutrient medium: the minimal inhibitory concentration (MIC) values of barium chloride ranged from 0.5 to 6 g/L, the MIC₉₀ value was 2 g/L. At the same time, 86.1% of all strains of fluorescent Pseudomonas species produced fluorescein on the control BaCl₂-free synthetic nutrient medium. For representatives of other genera grown on all the studied nutrient media, the MIC values of barium chloride ranged from 20 to 50 g/L. The proposed method for determination of the sensitivity of bacteria to barium ions using the synthetic nutrient medium with 6 g/L of barium chloride as a criterion for the classification of barium-sensitive strains to the genus Pseudomonas is suitable for standardization.     

Development of an accelerated method of determining the antibiotic sensitivity of Cl. perfringens type A]

An express method for determination of antibiotic sensitivity in the strains of Cl. perfringens of type A using Soviet dry nutrient media and antibiotics is proposed. The criteria for estimation of the level of the antibiotic sensitivity of the causative agent of gas gangrene in short periods on the basis of comparison of the data of the antibiotic agar diffusion procedure and the antibiotic MIC were worked out. Twelve antibiotics and 45 collection strains of Cl. perfringens of type A were used in the experiment. The antibiotic agar diffusion method with the use of the nutrient media, microbial load and cultivation conditions developed by the authors is recommended for tentative determination of the antibiotic sensitivity in Cl. perfringens of type A for 4 hours. The use of the agar diffusion method and determination of the antibiotic MIC provided complete estimation of tha antibiotic sensitivity of Cl. perfringens of type A within not more than 24 hours.     

A chitinolytic actinomycete complex in black soil

A chitinolytic actinomycete complex in chernozem soil has a specific taxonomic composition, which differs from that of the actinomycete complex which is typically isolated on standard nutrient media containing sugars and organic acids as carbon sources. The actinomycete complex that was isolated by using nutrient media with chitin as the source of carbon and nitrogen was dominated by representatives of the genus Streptosporangium, and the actinomycete complex that was isolated by using nutrient media with sugars and organic acids as the carbon sources was dominated by representatives of the genus Streptomyces. The confirmation to the ability of actinomycetes to utilize chitin as a sole source of carbon and nitrogen came from the augmented length and biomass of the mycelium, the increased number and biomass of the actinomycete spores, the production of carbon dioxide, and the accumulation of NH4+ ions in the culture liquid of the actinomycetes that are grown in the nutrient media with chitin.     

Consumption of leaf detritus by a stream shredder: Influence of tree species and nutrient status

Four species of riparian vegetation (alder, birch, willow and poplar) were fertilized with nitrogen, phosphorus, nitrogen + phosphorus, or no fertilizer (control). The resulting leaf detritus (leached but not microbially colonized) was offered to a stream shredder, Hydatophylax variabilis (Trichoptera: Limnephilidae). In one experiment, shredder consumption of leaf detritus from different nutrient treatments (within tree species) was compared, and in a second experiment, consumption of different tree species (within nutrient treatments) was compared. Larvae preferred leaf detritus from nitrogen + phosphorus treatments (except in poplar where nitrogen treatment was preferred). Alder was preferred over other tree species for all treatments. Chemical and physical analyses of leaf litter showed differences between tree species and nutrient treatments in nutrient content, tannins and leaf toughness. Leaf consumption by larvae was positively associated with nitrogen content and negatively associated with condensed tannin content. Species composition and nutrient status of riparian vegetation may strongly influence detrital food webs in streams.     

Cultivation of Pseudomonas aeruginosa in media with chlorella fermentative hydrolysate as nutrient base

The specific features of the growth of P. aeruginosa in media prepared with the use of Chlorella fermentative hydrolysate as nutrient basis were studied. The accumulation of microbial biomass was shown to depend on increased content of amino nitrogen in the media, while the effectiveness of the pyocyanin formation was linked with decreased content of amino nitrogen and a rise in the amount of sodium chloride.     

Microalgae for oil: strain selection, induction of lipid synthesis and outdoor mass cultivation in a low-cost photobioreactor

Thirty microalgal strains were screened in the laboratory for their biomass productivity and lipid content. Four strains (two marine and two freshwater), selected because robust, highly productive and with a relatively high lipid content, were cultivated under nitrogen deprivation in 0.6-L bubbled tubes. Only the two marine microalgae accumulated lipid under such conditions. One of them, the eustigmatophyte Nannochloropsis sp. F&M-M24, which attained 60% lipid content after nitrogen starvation, was grown in a 20-L Flat Alveolar Panel photobioreactor to study the influence of irradiance and nutrient (nitrogen or phosphorus) deprivation on fatty acid accumulation. Fatty acid content increased with high irradiances (up to 32.5% of dry biomass) and following both nitrogen and phosphorus deprivation (up to about 50%). To evaluate its lipid production potential under natural sunlight, the strain was grown outdoors in 110-L Green Wall Panel photobioreactors under nutrient sufficient and deficient conditions. Lipid productivity increased from 117 mg/L/day in nutrient sufficient media (with an average biomass productivity of 0.36 g/L/day and 32% lipid content) to 204 mg/L/day (with an average biomass productivity of 0.30 g/L/day and more than 60% final lipid content) in nitrogen deprived media. In a two-phase cultivation process (a nutrient sufficient phase to produce the inoculum followed by a nitrogen deprived phase to boost lipid synthesis) the oil production potential could be projected to be more than 90 kg per hectare per day. This is the first report of an increase of both lipid content and areal lipid productivity attained through nutrient deprivation in an outdoor algal culture. The experiments showed that this marine eustigmatophyte has the potential for an annual production of 20 tons of lipid per hectare in the Mediterranean climate and of more than 30 tons of lipid per hectare in sunny tropical areas.     

Production process in <Emphasis Type="Italic">Salicornia europaea</Emphasis> plants as a prospective phototrophic constituent in bioregenerative life support systems

Plant productivity of a common glasswort Salicornia europaea L. was investigated in relation to the type of nitrogen nutrition and as a function of macronutrient concentrations mimicking the mineral composition of human urine. The source of nutrient nitrogen had no substantial effect on productivity of Salicornia europaea. In plants grown on media with amide as a nitrogen source, the content of nitrogenous substances, including glutamic and aspartic amino acids, was higher than in plants grown with nitrate. In plants grown on media with mineral composition analogous to that of human urine, the shoots accumulated Na and K in almost equally high amounts, on the background of high and nearly equal Na and K concentrations in the nutrient media.     

接种丛枝菌根真菌对模拟大气氮沉降下灌木铁线莲根系形态及养分承载的影响

为了解菌根化处理的灌木铁线莲(Clematis fruticosa)苗木根系形态及养分承载对氮沉降的应激响应,以1年生盆栽灌木铁线莲为对象,分别采用单接种和混合接种,即：单接种根内根孢囊霉(Rhizophagus intraradices,以下简称+R),单接种摩西斗管囊霉(Funneliformis mosseae,以下简称+F);混合接菌(上述2菌种菌剂按体积1∶1混合,以下简称+RF)的菌根苗。以非菌根苗(未接菌,以下简称-M)为对照。氮沉降处理设置4个梯度(不施氮(0N,0 g·m^-2·a^-1)、低氮(LN,3 g·m^-2·a^-1)、中氮(MN,6 g·m^-2·a^-1)、高氮(HN,9 g·m^-2·a^-1)),1年后测定各处理细根形态(直径≤0.5 mm的总根长、总表面积、总体积、根尖数量)、菌根侵染率、土壤孢子密度及根、茎、叶各器官的养分(碳、氮、磷)含量等指标。(1)在+R和+RF处理下,LN处理的苗...     

Effect of sporulation and recovery medium on the heat resistance and amount of injury of spores from spoilage bacilli

AIMS: To assess the influence of sporulation media on heat resistance, and the use of stress recovery media to measure preservation injury of spores of five representative spoilage bacilli. METHODS AND RESULTS: Bacillus spores prepared on nutrient agar supplemented with Ca2+, Mg2+, Mn2+, Fe2+ and K+ were more heat-resistant than spores obtained from nutrient agar with Mn2+. This increased heat resistance correlated with a decrease in the protoplast water content as determined by buoyant density sedimentation. The degree of preservation injury severity could be assessed on media containing NaCl at moderate pH and organic acids at acid pH. Ca-DPA, K+ or proline were added to the recovery media to demonstrate that heat probably caused injury to both spore germination and the outgrowth system. SIGNIFICANCE AND IMPACT OF THE STUDY: The metal content of sporulation media can strongly effect the validity of preservation resistance studies. The distinctive recovery media developed here can be relevant for assessing and comparing new preservation technologies.     

三江平原小叶樟和毛果苔草中N素营养动态分析

讨论了沼泽湿地优势种小叶樟（Ｃａｌａｍａｇｒｏｓｔｉｓ ａｎｇｕｓｔｉｆｏｌｉａ）和毛果苔草（Ｃａｒｅｘ ｌａｓｉｏｃａｒｐａ）生物量和生长率变化情况,不同生长期各器官中Ｎ素含量及储量动态变化,以及植物对Ｎ素利用和区域养分限制情况。结果表明,两种植物地上生物量生长符合模式ｐ＝γ＋ａｔ＋βｔ＾２,地下生物量符合曲线ｐ＋ａ０＋ｂ０ｔ;受土壤水分、养分、气温和植物本身特点及其对Ｎ素选择吸收作用等多种因素     

氮素形态对黄檗幼苗生长及氮代谢相关酶类的影响

通过改变水培溶液中NH4+-N和NO3--N的比例, 研究了不同氮素形态对黄檗(Phellodendron amurense)幼苗生长及氮代谢相关酶类的影响。结果表明, 硝态氮比例较高的营养供给比铵态氮比例较高的营养供给有利于黄檗幼苗的生长, 叶片叶绿素含量和可溶性蛋白含量也高。在NH4+-N/NO3--N为25/75 时黄檗幼苗具有最大生物量。在铵态氮比例大的营养供给下, 黄檗幼苗的谷氨酰胺合成酶(GS)活性增强,而在硝态氮比例大的营养供给下幼苗的硝酸还原酶(NR)活性则较高, 叶片中的硝态氮较低。营养液的氮素形态及其组成通过影响GS与NR的活性而调控黄檗幼苗的氮素代谢。     

氮素形态对黄檗幼苗生长及氮代谢相关酶类的影响

通过改变水培溶液中NH4^＋-N和NO3^--N的比例,研究了不同氮素形态对黄檗（Phellodendron amurense）幼苗生长及氮代谢相关酶类的影响。结果表明,硝态氮比例较高的营养供给比铵态氮比例较高的营养供给有利于黄檗幼苗的生长,叶片叶绿素含量和可溶性蛋白含量也高。在NH4^＋-N／NO3^--N为25／75时黄檗幼苗具有最大生物量。在铵态氮比例大的营养供给下,黄檗幼苗的谷氨酰胺合成酶（GS）活性增强,而在硝态氮比例大的营养供给下幼苗的硝酸还原酶（NR）活性则较高,叶片中的硝态氮较低。营养液的氮素形态及其组成通过影响GS与NR的活性而调控黄檗幼苗的氮素代谢。     

Standardization of the nutrient medium for determining the biological activity of a number of antibiotics by the diffusion-in-agar method

The possibility of using 2 variants of a unified nutrient medium containing no Hottinger's broth for determination of the biological activity of a large number of antibacterial antibiotics was shown. The medium provided satisfactory reproducibility of the results. It contained yeast extract as a source of nitrogen. The medium was selected with the method of the fractional factor experiment. Clear inhibition growth zones and the slope of the dose-response curve were used as the criteria for estimation of the optimal medium. The described nutrient medium allowed a 2-time increase in the sensitivity of the method used for determination of the biological activity of neomycin, monomycin, streptomycin, oxytetracycline, chlortetracycline and erythromycin.     

Inference of allelopathy is complicated by effects of activated carbon on plant growth

Allelopathy can play an important role in structuring plant communities, but allelopathic effects are often difficult to detect because many methods used to test for allelopathy can be confounded by experimental artifacts. The use of activated carbon, a technique for neutralizing allelopathic compounds, is now employed in tests for allelopathy; however, this technique also could produce large experimental artifacts. In three independent experiments, it was shown that adding activated carbon to potting media affected nutrient availability and plant growth. For most species tested, activated carbon increased plant biomass, even in the absence of the potentially allelopathic agent. The increased growth corresponded to increased plant nitrogen content, likely resulting from greater nitrogen availability. Activated carbon also affected nitrogen and other nutrient concentrations in soil media in the absence of plants. The observed effects of activated carbon on plant growth can confound its use to test for allelopathy. The detection of allelopathy relies on the difference between plant growth in medium with carbon and that in medium without carbon in the presence of the potentially allelopathic competitor; however, this difference may be biased if activated carbon alters soil nutrient availability and plant growth even in the absence of the focal allelopathic agent.     

Anomalous potential response and (Na+ + K+)-ATPase in in vitro frog gastric mucosa

In general, increasing K+ on the nutrient side decreases the transmucosal PD (nutrient becomes more negative) but after bathing the mucosa in zero K+ media for about 30 min, or longer, elevation of K+ on the nutrient side increases the PD, an anomalous effect. In Cl- media, increasing nutrient K+ from zero to 4 mM produces an increase in PD (an anomalous response) of 3.1 and 5.3 mV in 2 and 5 min, respectively. Ouabain (10(-3) M) to the nutrient side abolished the anomalous response as did removal of Na+ (choline for Na+) from bathing media. In SO4(2-) media (SO4(2-) for Cl-), a significant anomalous PD response was observed when K+ on the nutrient side was increased from zero to 1, 2 or 3 mM but not to higher K+ concentrations. In this case, ouabain also abolished the anomalous response. It is postulated, on the basis of the effects of ouabain and the use of choline media, that an electrogenic (Na+ + K+)-ATPase pump is present on the nutrient-facing membrane in which more Na+ than K+ are transported per cycle.     

Normal and anomalous potential responses due to K+ changes in bullfrog antrum

The effect of changing the K+ concentration in the bathing media was studied in the bullfrog antrum. Usually increasing K+ on the nutrient side in standard Cl- -containing and Cl- -free solutions decreased the transmucosal potential difference (nutrient became more negative) - a normal effect. Similar results were obtained on the secretory side. Moreover, for K+ changes on the nutrient side in Cl- media, a plot of magnitude of delta V vs. log [K+] was linear for [K+] greater than 20 mM with slope of 27 mV per 10-fold change in [K+]. However, after bathing the mucosa in Cl- media with zero K+ for about 20 min, elevating the nutrient [K+] to 4 mM increased the potential difference (V) by 4.8 mV in 5 min and repeating the same sequence increased V by 6.9 mV in 5 min - both anomalous effects. Beyond 20 mM K+ the response was normal. In SO2-4 media, an anomalous potential difference of about 1 mV was obtained for changes from 0 to 3 or 6 mM nutrient K+. Ouabain (1 X 10(-3) M) in the nutrient solution abolished the anomalous response in Cl- and SO2-4 media. The normal response is attributed to passive, conductance pathways and the anomalous response because of the effect of ouabain, to a (Na+ + K+)-ATPase pump on the nutrient-facing membrane in which more Na+ than K+ ions are transported per cycle.     

中小型土壤动物组成对复垦铜尾地土壤养分的影响

通过盆钵生物试验研究了中小型土壤动物不同类群的组成对复垦铜尾矿废弃地土壤养分的影响。整个培养试验根据需求对不同中小型土壤动物组合设置四个处理措施,每种措施分为有无凋落物两种水平,即:(1)无土壤动物(Re),无土壤动物+凋落物(Re+Li)(2)跳虫(Co),跳虫+凋落物(Co+Li)(3)跳虫+蜱螨类(Co+Ac),跳虫+蜱螨类+凋落物(Co+Ac+Li)(4)中小型土壤动物(剔除大型土壤动物Me),中小型土壤动物+凋落物(Me+Li)。另设一组对照(CK)处理,并于培养实验开始前完成测定,作为本底值。分析表明,土壤p H随着中小型土壤动物类群的增加而降低,最低值出现在Me和Me+Li处理,且二者之间无显著性差异;对照(CK)处理下的各养分含量均显著低于其他各处理,各处理中有机质、有效磷和水解性氮的含量的最大值均出现Me+Li处理,且与其他各处理呈极显著差异(P0.01);土壤全磷含量最大值出现在Re和Me+Li处理,最低值出现在Co+Li处理且之间呈显著差异(P0.05);土壤全氮含量最大值出现在Co+Ac和Me+Li处理,最低值出现在Co+Ac+Li处理且彼此间也呈显著差异(P0.05)。相关分析表明,有机质与有效磷以及全氮与水解性氮之间相关性达到极显著正相关关系(r=0.886,r=0.898),有机质与全氮及水解性氮分别呈现显著正相关关系(r=0.735,r=0.780),全氮与有效磷(r=0.782)以及有效磷与水解性氮(r=0.755)也呈现显著正相关关系。p H与有机质呈现显著负相关关系(r=-0.782),全磷与有机质、全氮、有效磷、水解性氮均无相关关系。     

Efficient iron plaque formation on tea(Camellia sinensis) roots contributes to acidic stress tolerance

Tea plants grow in acidic soil, but to date, their intrinsic mechanisms of acidic stress tolerance have not been elucidated. Here, we assessed the tea plant response to growth on NHt4 nutrient media having different p H and iron levels. When grown in standard NHt4 nutrient solution(iron insufficient, 0.35 mg Là1 Fe2t), tea roots exhibited significantly lower nitrogen accumulation, plasma membrane Ht-ATPase activity, and protein levels; net Htefflux was lower at pH 4.0 and 5.0 than at pH 6.0. Addition of30 mg Là1 Fe2t(iron sufficient, mimicking normal soil Fe2tconcentrations) to the NHt4 nutrient solution led to more efficient iron plaque formation on roots and increased root plasma membrane Ht-ATPase levels and activities at p H 4.0 eland 5.0, compared to the p H 6.0 condition. Furthermore,plants grown at pH 4.0 and 5.0, with sufficient iron,exhibited significantly higher nitrogen accumulation than those grown at pH 6.0. Together, these results support the hypothesis that efficient iron plaque formation, on tea roots, is important for acidic stress tolerance. Furthermore,our findings establish that efficient iron plaque formation is linked to increased levels and activities of the tea root plasma membrane Ht-ATPase, under low pH conditions.