Antibacterial Action of Essential Oils of Artemisia as an Ecological Factor: I. Antibacterial Action of the Volatile Oils of Artemisia tridentata and Artemisia nova on Aerobic Bacteria

Bacterial response to increasing amounts of the volatile oils varies significantly according to species of bacteria tested. Among the four species examined, Escherichia coli was the most resistant to the oils, followed by Neisseria sicca, Bacillus subtilis, and Staphylococcus aureus. The oils of Artemisia tridentata seem to have the same degree of antibacterial action as oils obtained from A. nova.     

Comparison of Rumen Microbial Inhibition Resulting from Various Essential Oils Isolated from Relatively Unpalatable Plant Species

Essential oils were isolated from eight plant species which were relatively unpalatable to sheep and deer. The inhibitory potency of these essential oils upon sheep and deer rumen microorganisms was compared, in terms of total gas and volatile fatty acid (VFA) production, by use of an anaerobic manometric technique. Inhibitory effects of oils from the eight plant species may be placed in four groups: (i) essential oils from vinegar weed (Trichostema lanceoletum) and California bay (Umbellularia californica) inhibited rumen microbial activity most; (ii) lesser inhibition was exhibited by rosemary (Rosmarinus officinalis) and California mugwort (Artemisia douglasiana) oils, followed by (iii) blue-gum eucalyptus (Eucalyptus globulus) and sagebrush (Artemisia tridentata) oils; and (iv) oils from Douglas fir (Psuedotsuga menziesii) and Jerusalem oak (chenopodium botrys) resulted in the least inhibition, when 0.3 ml of each oil was used. A highly significant correlation coefficient (r = 0.98(**)) between total gas and VFA production indicated the validity of either method to measure the activity of rumen microorganisms. Our results are discussed in relation to the hypothesis that the selectivity and voluntary consumption of ruminants are related to the characteristic odor and antibacterial action of essential oils isolated from relatively unpalatable plant species.     

Effect of Various Essential Oils Isolated from Douglas Fir Needles upon Sheep and Deer Rumen Microbial Activity

The effects of essential oils isolated from Douglas fir needles on sheep and deer rumen microbial activity were tested by use of an anaerobic manometric technique. Rumen microorganisms were obtained from a sheep which had been fed mainly on alfalfa hay and dried range grass. One deer used in this study had access to Douglas fir trees the year around, whereas the other deer had no access to Douglas fir. All of the monoterpene hydrocarbons isolated from Douglas fir needles—α-pinene, β-pinene, limonene, myrcene, camphene, Δ³-carene, and terpinolene—promoted only slightly or had no effect on deer rumen microbial activity, whereas all of them promoted activity in sheep rumen microbes, except Δ³-carene and terpinolene, which inhibited activity. Of the oxygenated monoterpenes, all monoterpene alcohols—α-terpineol, terpinen-4-ol, linalool, citronellol, and fenchyl alcohol—strongly inhibited the rumen microbial activity of both sheep and deer. Monoterpene esters (bornyl acetate) produced mild inhibition for both sheep and deer microbes, and citronellyl acetate inhibited rumen microbial activity in sheep, whereas it promoted activity in both deer. Monoterpene aldehyde (citronellal) inhibited the activity of rumen microbes from both sheep and deer having no access to Douglas fir from the Hopland Field Station, whereas they produced no effect upon the deer having access to Douglas fir from the Masonite forest. Rumen microbial activity for sheep and deer was promoted slightly with aliphatic ester (ethyl-n-caproate). There was a marked difference between sheep and deer rumen microbes as affected by addition of the various essential oils. The monoterpene hydrocarbons promoted activity more on sheep rumen microbes than on deer, and the monoterpene alcohols inhibited sheep rumen microbial activity more than that of deer. Furthermore, the deer rumen microbes from Hopland Field Station were affected more than the deer from Masonite forest.     

Rumen Microbial Ecology in Mule Deer

Mule deer rumen microbial populations from animals in the natural habitat in Utah and from captive deer fed various rations were studied. The microorganisms were characterized on the basis of morphology and Gram reaction. Rumen samples contained 13 identifiable types of bacteria and one genus of ciliate protozoa (Entodinium). Highest rumen bacterial populations were produced on rations containing barley. No differences in proportions of ruminal bacteria in the various morphological groups could be detected when animals were fed either natural browse plants or alfalfa hay. The total numbers of bacteria were similar for animals feeding on controlled diets of browse or hay and those in the natural habitat. Numbers of some bacterial types were directly related to ciliate protozoal numbers, whereas others were inversely related. Highest rumen ciliate protozoal populations were observed on rations containing barley. No differences in protozoal populations were noted between diets containing only browse or hay. Seasonal variations were noted in ciliate protozoal numbers from deer feeding in the natural habitat. The total number of ciliate protozoa decreased in the fall and winter and remained low until spring. There were indications that salt in the deer diet favorably affected rumen ciliate protozoa. Rather than revealing direct deer management applications, this study serves to stimulate and illuminate new approaches to research in range and wildlife nutrition.     

种植青蒿与野生青蒿精油成分对比分析

用水蒸气蒸馏法和GC-MS法对来源于重庆云阳的栽培青蒿和野生青蒿精油的化学成分进行分析.从冬季和春季栽培的青蒿精油中分别鉴定出29和28个化合物,野生青蒿精油中鉴定出21个化合物.3种精油的化学成分差别明显,两种栽培青蒿精油有16个化合物相同.而栽培青蒿与野生青蒿精油仅有6个化合物相同.这表明基因型对青蒿精油的组成起决定作用;而气候因素对各成分的相对含量有重要影响,尤其对蒿酮和樟脑的积累影响较大.本文对进一步研究青蒿精油中萜类化合物之间的生源关系具有一定的参考价值.     

Antimicrobial Action of Some Citrus Fruit Oils on Selected Food-Borne Bacteria

The antimicrobial properties of essential oils, terpineol, and orange oil, in particular, varied according to the type of bacteria tested. Terpineol and other terpeneless fractions of citrus oils appeared to have greater inhibitory effect on food-borne bacteria than the other citrus oils or derivatives. Gram-positive bacteria were, in general, more sensitive to essential oils than gram-negative bacteria. Terpineol extended the shelf life of commercially pasteurized skim milk, low-fat milk, and whole milk for more than 56 days at 4 C. Orange oil extended the shelf life of skim milk and low-fat milk for the same period.     

獐的警戒行为模式及逃跑起始距离的适应性变化

为了掌握獐(Hydropotes inermis)的警戒行为特征并为重引入项目提供管理依据,以人为干扰源观察獐的警戒反应,发现其警戒模式包括听(hear)或扫视(scan)、盯视(stare)、走开(walk away)、跑开(run away)、吼叫(bark)和压脖(stretch)。利用逃跑起始距离对上海松江野化圈养(自主采食)獐、上海华夏圈养(人工饲喂)獐和江苏盐城野生獐警戒性进行比较,得出人工饲喂獐警戒性最小,野生獐警戒性最大。野化獐警戒性提高,表明可通过降低人类活动和种群密度、扩大区域面积等途径野化提高獐警戒性。     

Variation of the Chemical Composition and Antimicrobial Activity of the Essential Oils of Natural Populations of Tunisian Daucus carota L. (Apiaceae)

The essential oils of Daucus carota L. (Apiaceae) seeds sampled from ten wild populations spread over northern Tunisia were characterized by GC‐FID and GC/MS analyses. In total, 36 compounds were identified in the D. carota seed essential oils, with a predominance of sesquiterpene hydrocarbons in most samples (22.63–89.93% of the total oil composition). The main volatile compounds identified were β‐bisabolene (mean content of 39.33%), sabinene (8.53%), geranyl acetate (7.12%), and elemicin (6.26%). The volatile composition varied significantly across the populations, even for oils of populations harvested in similar areas. The chemometric principal component analysis and the hierarchical clustering identified four groups, each corresponding to a composition‐specific chemotype. The in vitro antimicrobial activity of the isolated essential oils was preliminarily evaluated, using the disk‐diffusion method, against one Gram‐positive (Staphylococcus aureus) and two Gram‐negative bacteria (Escherichia coli and Salmonella typhimurium), as well as against a pathogenic yeast (Candida albicans). All tested essential oils exhibited interesting antibacterial and antifungal activities against the assayed microorganisms.     

Chemical Polymorphism of Essential Oils of Artemisia vulgaris Growing Wild in Lithuania

Compositional variability of mugwort (Artemisia vulgaris L.) essential oils has been investigated in the study. Plant material (over ground parts at full flowering stage) was collected from forty‐four wild populations in Lithuania. The oils from aerial parts were obtained by hydrodistillation and analyzed by GC(FID) and GC/MS. In total, up to 111 components were determined in the oils. As the major constituents were found: sabinene, 1,8‐cineole, artemisia ketone, both thujone isomers, camphor, cis‐chrysanthenyl acetate, davanone and davanone B. The compositional data were subjected to statistical analysis. The application of PCA (Principal Component Analysis) and AHC (Agglomerative Hierarchical Clustering) allowed grouping the oils into six clusters. AHC permitted to distinguish an artemisia ketone chemotype, which, to the best of our knowledge, is very scarce. Additionally, two rare cis‐chrysanthenyl acetate and sabinene oil types were determined for the plants growing in Lithuania. Besides, davanone was found for the first time as a principal component in mugwort oils. The performed study revealed significant chemical polymorphism of essential oils in mugwort plants native to Lithuania; it has expanded our chemotaxonomic knowledge both of A. vulgaris species and Artemisia genus.     

Effective degradation of tannic acid by immobilized rumen microbes of a sika deer (Cervus nippon yesoensis) in winter

In winter seasons, wild sika deer (Cervus nippon yesoensis) inhabiting the Shiretoko Peninsula of Hokkaido Island, Japan, mainly graze woody materials (bark and twigs, etc.) as their feed source. Most of the tree species that they feed upon contain a high level of hydrolysable tannins within the inner bark. Tannins generally lead to low protein digestion and nutrient loss to these herbivorous mammals due to tannization of proteins. In winter months, it is speculated that wild sika deer develop a mechanism to degrade the tannins which are contained in their feed sources, but rumen fluid obtained from sika deer in winter months did not exhibit any ability to degrade tannins in liquid culture medium. However, constant degradation of hydrolysable tannin was observed when Ca-alginate gel beads were used for microbial immobilization and culturing. The gel beads that had been impregnated with 0.6×10⁴ fold-diluted rumen fluid of sika deer in winter and pre-incubated for 24 h under anaerobic conditions supplemented with a 1.5 g/L sugar were reacted with 5 g/L tannic acid solution. Under these conditions, the immobilized rumen bacteria grown in the macrogel beads effectively hydrolyzed tannic acid to release gallic acid monomers. Major bacterial colonies emerging in the Ca-alginate gel beads were identified as Streptococcus macedonicus and this bacterium (EC-D140) was regarded as the most likely candidate as the tannin-degrading bacterium.     

Glycosylated hemoglobin as a stable alternative to serum glucose in white-tailed deer

We compared serum glucose concentration and percent glycosylated hemoglobin (GH) in captive and wild white-tailed deer (Odocoileus virginianus) to determine stability of glucose relative to GH. Temporal changes in levels of serum glucose and GH were ascertained from serial blood samples collected from three captive deer over a 2-week period. State of glycemia also was determined for 17 wild deer that were collected from three populations in southeastern Oklahoma and southwestern Arkansas (USA). Concentration of serum glucose of captive deer decreased (P = 0.04) from 190.4 to 155.8 mg/dl over the 2 weeks; percent GH did not differ temporally (P = 0.30). Percent GH of wild deer did not differ (P = 0.23) when deer were separated into 2 groups (high and low state of glycemia) based on the median serum glucose concentration. We found a significant difference (P = 0.04) in percent GH among wild deer populations; serum glucose concentration did not differ (P = 0.72) among populations. Our results indicate that percent GH is more stable than serum glucose concentration and may be useful in population comparisons of nutritional condition.     

Acute sarcocystosis in a captive white-tailed deer in Virginia

A 19-mo-old female captive white-tailed deer in a public wild animal park in Richmond (Virginia, USA) was necropsied and evaluated histologically following spontaneous death after a 1 wk period of lethargy in a captive herd of 22 deer. An acute necrotizing pneumonia was associated with intraendothelial protozoal schizonts that were identified immunohistochemically as Sarcocystis spp. This is the first confirmed report of acute sarcocystosis in a wild ruminant.     

Insecticidal Activity and Chemical Composition of the Essential Oils of Artemisia lavandulaefolia and Artemisia sieversiana from China

In our screening program for new agrochemicals from local wild plants, Artemisia lavandulaefolia and A. sieversiana were found to possess insecticidal activity against the maize weevil Sitophilus zeamais. The essential oils of the aerial parts of the two plants were obtained by hydrodistillation and analyzed by GC and GC/MS. The main components of A. lavandulaefolia oil were caryophyllene (15.5%), β‐thujone (13.8%), eucalyptol (13.1%), and β‐farnesene (12.3%), and the principal compounds identified in A. sieversiana oil were eucalyptol (9.2%), geranyl butyrate (9.2%), borneol (7.9%), and camphor (7.9%). The essential oils of A. lavandulaefolia and A. sieversiana possessed fumigant toxicity against S. zeamais adults with LC₅₀ values of 11.2 and 15.0 mg/l air, respectively. Both essential oils also showed contact toxicity against S. zeamais adults with LD₅₀ values of 55.2 and 112.7 μg/adult, respectively.     

Bacteria and Methanogens Differ along the Gastrointestinal Tract of Chinese Roe Deer (Capreolus pygargus)

The current study provides the insight into the bacteria in the gastrointestinal tract (GIT) and methanogens presented in the rumen and cecum of the Chinese roe deer (Capreolus pygargus). The ruminal, ileal, cecal, and colonic contents, as well as feces, were obtained from each of the three, free-range, roe deer ingesting natural pasture after euthanasia. For the bacterial community, a total of 697,031 high-quality 16S rRNA gene sequences were generated using high-throughput sequencing, and assigned to 2,223 core operational taxonomic units (OTUs) (12 bacterial phyla and 87 genera). The phyla Firmicutes (51.2%) and Bacteroidetes (39.4%) were the dominant bacteria in the GIT of roe deer. However, the bacterial community in the rumen was significantly (P<0.01) different from the other sampled regions along the GIT. Secondly, Prevotella spp., Anaerovibrio spp., and unidentified bacteria within the families Veillonellaceae and Paraprevotellaceae were more abundant in the rumen than in the other regions. Unidentified bacteria within the family Enterobacteriaceae, Succinivibrio spp., and Desulfovibrio spp. were more predominant in the colon than in other regions. Unidentified bacteria within the family Ruminococcaceae, and Bacteroides spp. were more prevalent in the ileum, cecum and fecal pellets. For methanogens in the rumen and cecum, a total of 375,647 high quality 16S rRNA gene sequences were obtained and assigned to 113 core OTUs. Methanobrevibacter millerae was the dominant species accounting for 77.3±7.4 (S.E) % and 68.9±4.4 (S.E) % of total sequences in the rumen and cecum of roe deer, respectively. However, the abundance of Methanobrevibacter smithii was higher in the rumen than in the cecum (P = 0.004). These results revealed that there was intra variation in the bacterial community composition across the GIT of roe deer, and also showed that the methanogen community in the rumen differed from that in the cecum.     

Chemodiversity of Volatile Oils in Thapsia garganica L. (Apiaceae)

The chemical composition of the volatile oils obtained from the roots, leaves, flowers, and stems of Thapsia garganica of Tunisian origin was investigated by GC‐FID and GC/MS analyses. Sesquiterpene hydrocarbons and oxygenated monoterpenes were predominant in the oils of all plant parts. Bicyclogermacrene (21.59–35.09%) was the main component in the former compound class, whereas geranial (3.31–14.84%) and linalool (0.81–10.9%) were the most prominent ones in the latter compound class. Principal‐component (PCA) and hierarchical‐cluster (HCA) analyses revealed some common constituents, but also significant variability amongst the oils of the different plant parts. This organ‐specific oil composition was discussed in relation to their biological and ecological functions. For the evaluation of the intraspecific chemical variability in T. garganica, the composition of the flower volatile oils from four wild populations was investigated. Bicyclogermacrene, linalool, and geranial were predominant in the oils of three populations, whereas epicubenol, β‐sesquiphellandrene, and cadina‐1,4‐diene were the most prominent components of the oil of one population. PCA and HCA allowed the separation of the flower oils into three distinct groups, however, no relationship was found between the volatile‐oil composition and the geographical distribution and pedoclimatic conditions of the studied populations.     

圈养马麝的个性分类

本研究在甘肃兴隆山国家级自然保护区马麝繁育场展开,分别于2012年7月和2013年8月,针对374只圈养马麝,采用基于量表的个性评价法进行评价调查。结果表明:可建立包含17个个性项目的圈养马麝个性量表,因子分析显示每个项目公因子方差和载荷均大于0. 4,各因子累计贡献率为60. 33%,表明此量表具有较好的结构效度。此外Cronbach’sα系数为0. 787,表明量表具有较高的内部一致性信度,量表设计合理;因子分析和聚类分析结果显示,可将圈养马麝划分为五大个性维度:侵略性、社会性、刻板性、温顺性、活跃性。研究发现,圈养马麝个性维度与野生个体的个性类型划分存在差异,主要是受圈养条件和人为干扰等影响。圈养马麝个性量表和五大个性维度的建立对指导马麝繁育管理具有重要实践价值,填补了马麝个性研究的空缺,并为进一步进行麝类动物个性研究奠定了基础。另外,此类个性层次研究方法具有较好区分度,可为其他动物个性的初期研究提供参考。     

Chemical Composition,Cytotoxic and Antibacterial Activities of Essential Oils of Cultivated Clones of Juniperus communis and Wild Juniperus Species

Needles of seven cultivated clones (C1 – C7) of Juniperus communis at lower altitude and three wild Juniperus species (J. communis, J. recurva and J. indica) at higher altitudes were investigated comparatively for their essential oils (EOs) yields, chemical composition, cytotoxic and antibacterial activities. The EOs yields varied from 0.26 to 0.56% (v/w) among samples. Sixty‐one volatile components were identified by gas chromatography‐mass spectrometry (GC/MS) and quantified using gas chromatography GC (FID) representing 82.5 – 95.7% of the total oil. Monoterpene hydrocarbons (49.1 – 82.8%) dominated in all samples (α‐pinene, limonene and sabinene as major components). Principal component analysis (PCA) of GC data revealed that wild and cultivated Juniperus species are highly distinct due to variation in chemical composition. J. communis (wild species) displayed cytotoxicity against SiHa (human cervical cancer), A549 (human lung carcinoma) and A431 (human skin carcinoma) cells (66.4 ± 2.2%, 74.4 ± 1.4% and 57.4 ± 4.0%), respectively, at 200 μg/ml. EOs exhibited better antibacterial activity against Gram‐positive bacteria than against Gram‐negative bacteria with the highest zone of inhibition against Staphylococcus aureus MTCC 96 (19.2 ± 0.7) by clone‐7. As per the conclusion of the findings, EOs of clone‐2, clone‐5 and clone‐7 can be suggested to the growers of lower altitude, as there is more possibility of uses of these EOs in food and medicinal preparations.     

不同方法提取的荷叶挥发油化学成分分析

采取超临界CO2萃取和水蒸气蒸馏提取荷叶挥发油,利用GC-MS对它们进行了定性、定量分析。结果表明这两种方法提取荷叶挥发油的化学成分及含量皆有很大差别。超临界CO2萃取的荷叶挥发油更具天然性,超临界CO2萃取法为提取荷叶挥发油的理想方法。     

不同方法提取的艾叶挥发油指纹图谱分析

采取超临界CO2、微波和水蒸气蒸馏三种方法提取艾叶挥发油,利用GC-MS分析了不同方法提取的 艾叶挥发油指纹图谱。超临界CO2和微波萃取的艾叶挥发油的化学成分相似,水蒸气蒸馏与前两种方法提 取的挥发油的化学成分有些差异。超临界CO2萃取的艾叶挥发油更具天然性,超临界CO2萃取法为提取艾 叶挥发油的较理想方法;微波辅助萃取也不失为一种可行的方法。     

Genetic assessment of captive Chinese water deer populations in Zhoushan Archipelago,China

Chinese water deer is a rare and vulnerable animal in China because of the poaching for medical use and the habitat loss. In this study, the genetic diversity and population genetic structure of 40 Chinese water deer from three populations in Zhoushan Archipelago were investigated with ten highly polymorphic microsatellite loci, including 4 screened from the nuclear DNA in the study, and 6 selected from the literature. According to the results, these captive populations had a higher genetic diversity than other rare cervid species, such as forest musk deer. No signs of inbreeding were detected. Low genetic differentiation among these populations was found. The probable reasons included the isolation by distance, the exchange among islands, or the supplement of the wild Chinese water deer. We proposed the deer raisers to strengthen the exchanges from different islands or mainland, and if possible, some deer would be returned to the wild to expand the wild population.