Ethanol extracts (AF-06, 07, and 08, 10 mg/kg) of Brazilian propolis were administered orally to cutaneously herpes simplex virus type 1 (HSV-1)-infected mice three times daily on days 0 to 6 after infection to evaluate their efficacies against HSV-1 infection and significantly limited development of herpetic skin lesions. AF-07 and 08 significantly reduced virus titers in brain and/or skin on day 4 without toxicity, but AF-08 had no anti-HSV-1 activity in vitro. AF-06 and 08 significantly enhanced delayed-type hypersensitivity (DTH) to inactivated HSV-1 antigen in infected mice. Oral AF-08-administration significantly augmented interferon (IFN)-γ production by HSV-1 antigen from splenocytes of HSV-1-infected mice, while direct exposure of splenocytes of infected mice to AF-06 significantly elevated IFN-γ production in vitro. Thus, AF-08 might have components that are active in vivo even after oral administration and those of AF-06 might be active only in vitro. Because DTH is a major host defense for intradermal HSV-1 infection, augmentation of DTH response by AF-06 or 08, directly or indirectly, respectively, may contribute to their efficacies against HSV-1 infection. In addition, AF-06 and 07 possibly contain anti-HSV-1 components contributing to their efficacies. Such biological activities of Brazilian propolis may be useful to analyze its pharmacological actions.

The effects of a dried Bacillus subtilis culture on the egg qualities of layers were studied. Nine hundred and sixty 25-wk-old Lohmann Brown layers were randomly divided into 5 groups with 192 layers in each group. Layers in group 1 were fed a control diet. The remaining groups received the control diet that contained either 20 mg of zinc bacitracin/kg and 4 mg of colistin sulfate/kg or 500, 1,000, or 1,500 mg of B. subtilis culture/ kg, respectively. The results showed improvements in egg production, feed consumption, and feed conversion (P < 0.05) of layers when 500 mg of B. subtilis culture/kg was added to the diets. The results also showed some special improvements in this group, including increases in eggshell thickness, yolk color, and Haugh unit, and decreases in yolk cholesterol concentration (P < 0.05). However, excessive doses of B. subtilis culture did not improve the performance of layers.

Cucumis sativus L., which belongs to the Cucurbitaceae family, is cultivated worldwide as a vegetable crop. The fruit of C. sativus (cucumber) is used not only as food, but also in folk medicines and folk cosmetics. The leaves, stems and roots of C. sativus are reportedly used in Chinese folk medicine as antidiarrheal, detoxicant and antigonorrheal agents. 1)However, these are not major uses, so after harvest of the cucumber fruits, almost all the remaining plant material is waste. There have been few chemical studies regarding the constituents of this material. The only reports are by Enslin et al., who isolated the triterpenoid cucurbitacin C, 2) and McNally et al., who isolated C-glycosyl flavonoid.3) Thus, in the present study, we attempted to identify useful components of cucumber plant material to support its development as a medicinal resource.As part of our ongoing search for medicinal materials in cucumber plant material, chromatographic separation and purification led to the identification of two new megastigmanes, one known megastigmane, one known carotenoid, one known lignan and three other known compounds. In the present paper, we report the isolation and structural elucidation of the new megastigmanes. Results and DiscussionAir-dried leaves of C. sativus were extracted using methanol (MeOH). The MeOH extract was partitioned into an ethyl acetate (EtOAc)-water mixture to furnish EtOAcsoluble and water-soluble parts. The EtOAc-soluble part was subjected to normal-and reversed-phase column chromatography and HPLC to yield compounds 1, 3, 4, 5, 6 and 7 (Fig. 1). The water-soluble part was again extracted with EtOAc, then with n-butanol (BuOH), using an automatic extractor. The n-BuOH-soluble part was subjected to normal-and reversed-phase column chromatography and HPLC to yield compounds 2 and 8 (Fig. 1).Compound 1 C heteronuclear multiple bond coherence (HMBC) study for 1, long-range correlations were observed between the following protons and carbons: H-4 and C-2; H-2a, 2b and C-3; H-13 and C-4; H-2a, 2b, 4, 7, 8, 13 and C-6; H-7, 8 and C-9; and H-10a, 10b and C-8 (Fig. 2). Furthermore, acetylation of 1 with acetic anhydride in dry pyridine yielded di-O-acetate (1a), whose 13 C-NMR spectrum showed an acetylation shift around the 9 and 10 position of 1a. These findings and comparisons of the 1 H-and 13 C-NMR spectra of 1 and 1a with those of a known megastigmane led us to formulate the 3-oxo-6,9,10-trihydroxy-ionone structure of 1 (Fig. 1). The absolute configuration of C-6 of 1 was determined by application of the circular dichroism (CD) helicity rule 4) with known (ϩ)-dehydrovomifoliol (3). The CD spectrum of 3 showed a positive Cotton effect (De ϩ27.00) at 243.2 nm, and the 6-position was determined to have an S configuration by the CD helicity rule.4) On the basis of this evidence, the CD spectrum of 1 exhibited a positive Cotton effect (De ϩ15.79) at 240.8 nm, and the 6-position was shown to have an S configuration. In order to clarify the absolute stereostructure of the 9-position of 1, a...

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Gut microbiota of grass carp plays an important role in host. However, detailed information regarding the changes of microbiota after probiotics administration in relation to the gastrointestinal microbiota is absent. In the present study, dietary administration of putative probiotics Shewanella xiamenensis A-1, Aeromonas veronii A-7, and Bacillus subtilisstrains was conducted in grass carp to investigate if there is a discernible alteration in intestinal microbiota and whether the alteration is associated with previous study about the immunity regulation in grass carp. Bacterial 16S rRNA gene sequence-based comparisons of the bacterial communities in the grass carp intestine were detected after 28 days feeding by five diets, and results demonstrated the changes of microbial community composition at genus level. The abundance of Cetobacterium genus with potential immunity function increased. Potential pathogens and probiotics are important constitutions of the intestinal microbiota. Orally taken probiotics considerably reduced the abundance of the potential pathogenic bacteria (e.g., Pseudomonas and Flavobacterium genus) in the intestine. Meanwhile, putative probiotics used in this study were favorable to the reproduction of potential probiotics in THE intestine of grass carp (e.g., Vibrio, Streptococcus, and Enterococcus genus). Moreover, modulation of intestinal environment by the probiotics could impact the abundance of cellulose-degrading bacteria (e.g., Citrobacter genus). Those results suggested that oral probiotics administration can positively improve the composition of intestinal microbial community in grass carp, and this was associated with regulation of immunity in grass carp. Probiotics-induced alteration of microbiota may potentially lower the risk of disease outbreaks during cultivation stage of grass carp.

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