Evaluation of in vitro antibacterial effect of essential oil and some herbal plant extract used against mastitis pathogens.

BACKGROUND: Mastitis in dairy cattle is a highly prevalent infectious disease, caused by various pathogens, mainly Staphylococcu aureus and Escherichia Coli, considerable economic loss worldwide. OBJECTIVES: The aim of this study was to evaluate the in vitro activity of Herbal plants used against S. aureus and E. coli bacteria which are the causative agents of mastitis. METHODS: Therefore, in this study we investigate the antimicrobial effect of plant to evaluate the in vitro antibacterial activity of squaw mint (Mentha pulegium L., Lamiaceae family), catnip (Nepeta cataria L., Lamiaceae), lemon balm (Melissa officinalis L., Lamiaceae), for mastitis treatment. Solutions prepared in fixed oils, against S. aureus and E. coli bacteria which are the main agents of mastitis. Isolation and antibiotic susceptibility analyses of milk samples taken from 100 subclinical mastitis dairy cows were performed. The antibacterial properties of the solutions were analysed by a disk diffusion method. RESULTS: In the bacterial isolation, S. aureus was determined 97.7% and E. coli 53.5% positive of cows with mastitis. Antibacterial susceptibility test of the Lemon balm extract and essential oil showed maximum zone of inhibition against S. aureus 30 µl (23 mm), followed by 20 µl (19 mm), E. coli (19 mm) and 10 µl (5-7 mm), of the same extract against the Gram-positive bacteria. The ethanol extracts show the similar activity against the Gram-negative bacteria at 30, 20, and 10 µl (18-20 mm). Followed by S. aureus, when the zone areas for the susceptible solutions (Lemon balm, and essential oil) and the control group were compared, determined that there was little difference between for S. aureus and E. coli. CONCLUSIONS: This study hence indicated that in vitro cultured plantlets of lemon balm and peppermint oil can be used as the alternative method for production of mastitis and cheap source its precursor with antimicrobial activities.

Cangpu Oral Liquid as a Possible Alternative to Antibiotics for the Control of Undifferentiated Calf Diarrhea.

Antibiotics are essential in the prevention of calf diarrhea epidemics. As more antibiotics become ineffective due to drug-resistant bacteria, attention must be directed toward alternative treatments for calf diarrhea. Natural antibiotic alternatives, such as Chinese herbal medicine, have become a research hotspot in the clinical treatment of diseases such as calf diarrhea due to their characteristics of fewer side effects, low cost, little residue, and no drug resistance. The Cangpu Oral Liquid (CP) was modified from a traditional herbal formula that had been widely used in ancient China to treat gastrointestinal diseases in animals. In order to evaluate the treatment effect of CP on neonatal calf diarrhea, a randomized controlled field trial was performed. Two hundred and forty-six diarrheal Holstein calves of 2-15 days old were selected and randomly divided into two treatment groups receiving either apramycin or CP. 101 out of 123 calves recovered from diarrhea in the CP group, whereas 77 out of 123 calves showed recovery after antibiotic therapy. There were no differences in initial weight between both groups, while the final weight was significantly different (P = 0.892, P = 0.025, respectively). The mean average daily gain (ADG) of calves (211.45 gram/day) in the CP group was significantly higher compared to the antibiotic group (164.56 gram/day) (P = 0.001). The CP group also showed a shorter recovery time from diarrhea (3.90 days vs. 6.62 days, P = 0.001). The current results indicate that the CP has a beneficial clinical effect on the treatment of diarrhea in neonatal calves and is an effective alternative treatment option.

A high-value-added application of the stems of Rheum palmatum L. as a healthy food: the nutritional value, chemical composition, and anti-inflammatory and antioxidant activities.

Rhubarb has edible stems or stalks. In this paper, we investigated the nutritional value, chemical composition, and bioactivities of Rheum palmatum stems (SRP) and analyzed the mode of action. SRP exhibited biosafety and had nutritional value, with abundant essential amino acids and minerals. Based on network pharmacology and western blot tests, we found that it showed anti-inflammatory activity via the PI3K-Akt-mediated NF-κB pathway. Out of 20 compounds identified using UPLC-ESI-Q-TOF/MS analysis, cirsiliol and hydrangenol were active compounds and they inhibited NO production in RAW264.7 cells induced by LPS. The alleviation of an inflammatory response is combined with a decrease in oxidative stress, and SRP showed antioxidant activity via attenuating antioxidant enzymes, scavenging free radicals, improving the mitochondrial membrane potential, and decreasing the reactive oxygen species level. These results indicated that SRP, with abundant flavonoids and a good nutritional composition, could be used as a dietary supplement for food applications.

Comparative study of antimicrobial action of aloe vera and antibiotics against different bacterial isolates from skin infection.

Aloe vera is reputed to have medicinal properties. For centuries, it has been used for an array of ailments such as mild fever, wounds and burns, gastrointestinal disorders, diabetes, sexual vitality and fertility problems to cancer, immune modulation, AIDS and various skin infections. In present study, antibacterial activity of aloe vera gel extracts was tested against some common skin infection pathogens, that is, Escherichia coli, Shigella, Salmonella spp. and Staphylococcus aureus all were recorded positive. Antibiotic resistance and susceptibility pattern of above isolates were also studied against 10 clinically significant antibiotics (ampicillin [AMC], amoxicillin, augmentin, cefotaxime, ceftazidime [CAZ], cefuroxime [CXM], ciprofloxaci, tetracycline, cefpodoxime and imipenem). AMC and CXM were found to be most effective antibiotic followed by CXM with highest efficacy against Gram-negative bacteria. In case of CAZ showed highest efficacy was showed against Gram-positive bacteria. Aloe vera leave gel was extracted with four different solvent-like aloe vera leaf extract, root extract, leaf ethanol extract and root ethanol extract; however, Gram-negative as well Gram-positive isolates was found highest susceptibility with aloe leaf and aloe root ethanol extract. Moderate sensitivity observed with aloe leaf extract and aloe root extract against both Gram-positive as well as Gram-negative bacterial isolates. This result showed that ethanol extracts of aloe vera both leaf and root can be used alongside conventional antibiotics to fight agents of infections that are so prevalent in the skin infection.

In Vitro and In Vivo Activities, Absorption, Tissue Distribution, and Excretion of OBP-4, a Potential Anti-Clostridioides difficile Agent.

Clostridioides difficile infection (CDI) is considered a major concern of the health care system globally, with an increasing need for alternative therapies. OBP-4, a new oxazolidinone-fluoroquinolone hybrid with excellent in vitro activities and good safety, shows promising features as an antibacterial agent. Here, we further evaluated the in vitro and in vivo activities of OBP-4 against C. difficile and its absorption (A), distribution (D), and excretion (E) profiles in rats. In vitro assays indicated that OBP-4 was active against all tested C. difficile strains, with MICs ranging from 0.25 to 1 mg/liter. In addition, OBP-4 showed complete inhibition of spore formation at 0.5× MIC. In the mouse model of CDI, 5-day oral treatment with OBP-4 provided complete protection from death and CDI recurrence in infected mice. However, cadazolid (CZD) and vancomycin (VAN) showed less protection of infected mice than did OBP-4 in terms of diarrhea and weight loss, especially VAN. Subsequently, ADE investigations of OBP-4 with a reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method showed extremely low systemic exposure and predominantly fecal excretion, resulting in a high local concentration of OBP-4 in the intestinal tract-the site of CDI. These results demonstrated that OBP-4 possesses good activity against C. difficile and favorable ADE characteristics for oral treatment of CDI, which support further development of OBP-4 as a potential anti-CDI agent.

Multiple Biological Activities of Rhododendron przewalskii Maxim. Extracts and UPLC-ESI-Q-TOF/MS Characterization of Their Phytochemical Composition.

Backgroud: Rhododendron przewalskii Maxim. is an evergreen shrub that is used as a traditional medicine in China. However, the modern pharmacology and the chemical components of this plant has not been studied. In this paper, we aimed to investigate the antifungal, anti-inflammatory and antioxidant activities and underlying mechanism of its aqueous and ethanol extracts, and analyze their chemical composition and active compounds of R. przewalskii. Methods: The antifungal activity was determined in vitro, and anti-inflammatory and antioxidant activities and underlying mechanism of its aqueous and ethanol extracts were evaluated in vitro and in RAW 264.7 cells. The chemical composition were analyzed using UPLC-ESI-Q-TOF/MS, and the contents of six compounds were determined via HPLC. Results: Both extracts of R. przewalskii showed promising anti-inflammatory activity in vitro; decreased the production of four inflammatory cytokines, namely, nitric oxide, IL-1ß, IL-6 and TNF-ɑ, in RAW 264.7 cells induced by lipopolysaccharide; and exhibited weak cytotoxicity. The extracts significantly scavenged DPPH radicals, superoxide radicals and hydroxyl radicals to exert antioxidant effects in vitro. The two extracts also exhibited cellular antioxidant activity by increasing superoxide dismutase and CAT activities and decreasing malondialdehyde content in RAW 264.7 cells induced by LPS. However, the antifungal activity of the two extracts was weak. Nine flavonoids were identified by UPLC-ESI-Q-TOF/MS. Of these, six compounds were analyzed quantitatively, including avicularin, quercetin, azaleatin, astragalin and kaempferol, and five compounds (myricetin 3-O-galactoside, paeoniflorin, astragalin, azaleatin and kaempferol) were found in this species for the first time. These compounds demonstrated antioxidant activities that were similar to those of the R. przewalskii extracts and were thought to be the active compounds in the extracts. Conclusion: R. przewalskii extracts presented promising anti-inflammatory and antioxidant activities. The extracts contained amounts of valuable flavonoids (8.98 mg/g fresh material) that were likely the active compounds in the extract contributing to the potential antioxidant activity. These results highlight the potential of R. przewalskii as a source of natural antioxidant and anti-inflammatory agents for the pharmaceutical industry.

A value-added application of eugenol as acaricidal agent: The mechanism of action and the safety evaluation.

Introduction: Eugenol is a major component of essential oils of several plants, it exhibits significant antiparasitic and acaricidal activities, yet its molecular targets remain unknown. Objectives: We aimed to systematically investigate the mechanism of action and the potential targets of eugenol against P. cuniculi, and evaluate the safety for laying the theoretical foundation for clinical application as an acaricide. Methods: Using RNA-Seq analysis, surface plasmon resonance analysis and RNA interference assay, the mode of action of eugenol against Psoroptes cuniculi was investigated. The effect on the mitochondrial membrane potential and complex I of PC12 cells and C6/36 cells was assayed to investigate the species specificity of eugenol in insects and mammals. Finally, a safety evaluation of eugenol in vivo was performed. Results: Eugenol inhibited complex I activity of the mitochondrial respiratory chain in the oxidative phosphorylation pathway by binding to NADH dehydrogenase chain 2 and resulted in the death of mites. The inhibition rates were 37.89% for 50 µg/mL and 60.26% for 100 µg/mL, respectively. Further experiments indicated that the difference in the complex I sequence between insects and mammals led to the different affinity of eugenol to specific peptide, resulting in species specificity. Eugenol exhibited significant inhibitory effects against the mitochondrial membrane potential and complex I in Aedes albopictus C6/36 cells but was not active in rat PC12 cells. Insect cells were particularly sensitive to eugenol. In contrast to the known inhibitor rotenone, eugenol had better safety and did not result in Parkinson's disease or other diseases in rats. Conclusion: This is the first report on acaricidal eugenol targeting complex I of the mitochondrial respiratory chain. This work lays the foundation for the development of eugenol as an environmentally alternative acaricidal agent.

The acaricidal mechanism and active compounds against Psoroptes cuniculi of the methanol extract of Adonis coerulea Maxim II: Integrated proteomics and SPR analysis.

Adonis coerulea Maxim. as a folk medicine, presented acaricidal acitvity. However, the mode of action and active compounds were unclear. In this study, using proteomics and surface plasmon resonance (SPR) technology the mode of action and active compounds of A. coerulea were investigated, as well as a sensitive and environmentally friendly analytical method developed. Proteomics analysis results showed that after treatment of mites with A. coerulea methanol extract (MEAC), 135 proteins were differentially expressed, most of them enriched in the myocardium pathway and participated in the function of the inflated cystic organ. Na+-K+-ATPase may be a potential target. Then, it was used as a target to capture the compounds from the extract using a SPR test. Twelve compounds were found, five compounds, namely ellagic acid, ouabain, convallatoxin, strophanthidin and cymarin presented the higher affinity with Na+-K+-ATPase in molecular docking test. Further study showed that the latter four compounds presented the stronger cytotoxicity and the inhibitory effect on Na+-K+-ATPase with IC50 values ranging with 2.38-0.56 µg/mL, and also showed toxicity against Psoroptes cuniculi. These results indicated that MEAC presented toxicity against mites by inhibiting Na+-K+-ATPase, and cardiac glycosides may be active compounds of this plant in terms of its acaricidal activity. Only 10 g of plant was used to investigate its active compounds. This study lays the foundation for developing sensitive methods for active compound detection.

The active compounds and AChE inhibitor of the methanol extract of Adonis coerulea maxim against Psoroptes cuniculi.

Adonis coerulea Maxim. presents acaricidal activity in vitro and in vivo, and inhibits AChE and other enzymes activities. However, the active compounds against Psoroptes cuniculi were still unclear. AChE, a common acaricidal and insecticidal target, plays a key role in neural conduction of mites. In this study, using surface plasmon resonance (SPR) technology, AChE was used as a target to capture the compounds from A. coerulea methanol extract (MEAC). After calculating the affinity with molecular docking, the inhibitory effect of compounds against AChE was studied. Results showed that 27 compounds were captured by AChE and identified from MEAC by LC-MS/MS. Among of these compounds, eight compounds presented the high affinity with AChE and high scores in molecular docking assay, especially for silibinin (-12.19 kcal/mol) and vitexin (-11.72 kcal/mol). Further studies showed that although these compounds have the weak cytotoxicity against C6/36 cells, silibinin, quercetin and corilagin could inhibit AChE activity with IC50 values of 40.11 µg/mL, 46.15 µg/mL and 50.98 µg/mL, respectively. These results indicated that silibinin, quercetin and corilagin may be responsible for AChE inhibition which contributes to the acaricidal properties of A coerulea. This study lays the foundation for developing sensitive and sustainability methods for active compound detection from plants.

Estimating 24-Hour Sodium Excretion from Spot Urine Samples in Chinese Adults: Can Spot Urine Substitute 24-Hour Urine Samples?

Several estimating equations for predicting 24-h urinary sodium (24-hUNa) excretion using spot urine (SU) samples have been developed, but have not been readily available to Chinese populations. We aimed to compare and validate the six existing methods at population level and individual level. We extracted 1671 adults eligible for both 24-h urine and SU sample collection. Mean biases (95% CI) of predicting 24-hUNa excretion using six formulas were 58.6 (54.7, 62.5) mmol for Kawasaki, -2.7 (-6.2, 0.9) mmol for Tanaka, -24.5 (-28.0, -21.0) mmol for the International Cooperative Study on Salt, Other Factors, and Blood Pressure (INTERSALT) with potassium, -26.8 (-30.1, -23.3) mmol for INTERSALT without potassium, 5.9 (2.3, 9.6) mmol for Toft, and -24.2 (-27.7, -20.6) mmol for Whitton. The proportions of relative difference >40% with the six methods were nearly a third, and the proportions of absolute difference >51.3 mmol/24-h (3 g/day salt) were more than 40%. The misclassification rate were all >55% for the six methods at the individual level. Although the Tanaka method could offer a plausible estimation for surveillance of the population sodium excretion in Shandong province, caution remains when using the Tanaka formula for other provincial populations in China. However, these predictive methods were inadequate to evaluate individual sodium excretion.

Magnetic molecularly imprinted polymers doped with graphene oxide for the selective recognition and extraction of four flavonoids from Rhododendron species.

Herein, a novel magnetic molecularly imprinted polymer doped with reticular graphene oxide (Fe3O4@SiO2-GO@MIPs) was synthesized for the selective recognition and extraction of 4 flavonoids (farrerol, taxifolin, kaempferol, and hyperin) from Rhododendrons species. The Fe3O4@SiO2-GO@MIPs with lamellar membranes showed outstanding adsorption capacity. The 3D cavities complementary to the "shape" of farrerol were "imprinted" on the polymer framework after removal of farrerol template. Competitive binding assays showed that the polymer has a higher selectivity for farrerol compared with other analogues and references. The Fe3O4@SiO2-GO@MIPs as solid-phase extraction adsorbents combined with liquid chromatography-tandem quadrupole mass spectrometry (LC-MS/MS) was used for selective determination of four flavonoids from Rhododendrons samples. The limits of detection (LOD) were 0.07, 0.08, 0.06, and 0.08 µg L-1 for farrerol, taxifolin, kaempferol, and hyperin, respectively. These results suggest that the prepared Fe3O4@SiO2-GO@MIPs have the potential applicability to extract, purify, and enrich flavonoids from herbs, supplements, and other natural products.

The Genus Adonis as an Important Cardiac Folk Medicine: A Review of the Ethnobotany, Phytochemistry and Pharmacology.

The genus Adonis L. (Ranunculaceae), native to Europe and Asia, comprises 32 annual or perennial herbaceous species. Due to their cardiac-enhancing effects, Adonis spp. have long been used in European and Chinese folk medicine. These plants have been widely investigated since the late 19th century, when the cardiovascular activity of Adonis vernalis L. was noted in Europe. The present paper provides a review of the phytochemistry, biological activities and toxicology in order to highlight the future prospects of the genus. More than 120 chemical compounds have been isolated, with the most important components being cardiac glycosides as well as flavones, carotenoids, coumarins and other structural types. Plants of the genus, especially A. vernalis L. and A. amurensis Regel & Radde, their extracts and their active constituents possess broad pharmacological properties, including cardiovascular, antiangiogenic, antibacterial, antioxidant, anti-inflammatory and acaricidal activities, and exhibit both diuretic effects and effects on the central nervous system. However, most plants within the 32 species have not been comprehensively studied, and further clinical evaluation of their cardiovascular activity and toxicity should be conducted after addressing the problem of the rapidly decreasing resources. This review provides new insight into the genus and lays a solid foundation for further development of Adonis.

The Anti-diarrheal Activity of the Non-toxic Dihuang Powder in Mice.

Dihuang powder (DHP) has been used in the traditional Chinese medicine for the treatment of diarrhea in some regions of China. But up to now, the anti-diarrheal activity of DHP haven't been performed with modern pharmacological technology. This study aims to investigate the quality control, the potential toxicity and anti-diarrheal activity of Dihuang powder in mice. High performance liquid chromatography (HPLC) and thin layer chromatography (TLC) were used to detect five active compounds in DHP for quality control, and the acute toxicity and sub-acute toxicity for 28-day oral administration of DHP were then evaluated. The anti-diarrheal activity was investigated using mouse model. Results showed that the levels of quercetin and berberine in DHP were 0.054 and 0.632 mg/g, respectively, and atractylodin, matrine, and patehouli aleohal were also detected in DHP. At the given doses, DHP was safe in terms of acute and sub-acute toxicity. Meanwhile, DHP exhibited strong anti-diarrheal effects as well as decreased gastrointestinal motility and the secretions induced by Sennae and castor oil in a dose-dependent manner. It could decrease the content of IL-1ß, IL-6, and TNF-α in the small intestine, and improve the histopathological changes of small intestine and large intestine induced by Sennae. The antinociceptive and anti-inflammatory activities in vivo also were presented. Based on all of the results, we thought that DHP has anti-diarrheal activity, and could be used to treat diarrhea as well as alleviate the pain and inflammation induced by diarrhea. This study provides a theoretical basis for the clinical use of DHP and may assist in the development of new drugs for the treatment of diarrhea. The mechanism of the anti-diarrheal activity should be investigated in the future.

Analysis of bauxite residue components responsible for copper removal and related reaction products.

Bauxite residue is a solid waste produced during alumina production process, and the storage of that in China reached 0.6 billion tons with an increase of more than 70 million annually. Bauxite residue can be used to remove heavy metals from water. This study analyzed components of bauxite residue responsible for copper removal, removal process and accompanying reaction products. Calcite (CaCO3), hematite (Fe2O3) and sulfur-Fe are main components contributing to copper removal. Sulfur in bauxite residue works with iron to remove copper. All these components reacted with copper immediately as bauxite residue was added. Reaction time of sulfur-Fe and carbonate was 5 min and 1 h, respectively. And hematite reacted until complete removal of copper (>2 h). Sulfur quickly reacted with coexisting iron to remove copper, producing chalcopyrite (CuFeS2), cubanite (CuFe2S3) and bornite (Cu5FeS4). Carbonate in bauxite residue reacted with copper, producing tenorite (CuO), copper hydroxide (Cu(OH)2), malachite (Cu2(OH)2CO3), carbonate cyanotrichite (Cu4Al2(CO3,SO4)(OH)12·2H2O), chalconatronite (Na2Cu(CO3)2·3H2O), nakauriite (Cu8(SO4)4(CO3)(OH)6·48H2O) and callaghanite (Cu2Mg2(CO3)(OH)6·2H2O). Copper precipitated through reaction with hematite to produce delafossite (CuFeO2). After removal reaction, the existing forms of copper in bauxite residue comprised carbonate-bound (73.6%-85.7%), iron oxide-bound (5.6%-23.8%), organic matter/sulfide-bound (0.5%-9.0%) and residual forms (0.9%-2.0%). In conclusion, removal of copper using bauxite residue features a more complex reaction than adsorption.

Coumarin Content, Morphological Variation, and Molecular Phylogenetics of Melilotus.

Melilotus albus and Melilotus officinalis are widely used in forage production and herbal medicine due to the biological activity of their coumarins, which have many biological and pharmacological activities, including anti-HIV and anti-tumor effects. To comprehensively evaluate M. albus and M. officinalis coumarin content (Cou), morphological variation, and molecular phylogeny, we examined the Cou, five morphological traits and the molecular characterization based on the trnL-F spacer and internal transcribed spacer (ITS) regions of 93 accessions. Significant (p < 0.05) variation was observed in the Cou and all five morphological traits in both species. Analysis of population differentiation (Pst) of the phenotypic traits showed that powdery mildew resistance (PMR) had the greatest Pst, meaning that this trait demonstrated the largest genetic differentiation among the accessions. The Pst values of dry matter yield (DMY) and Cou were relatively high. Biplot analysis identified accessions with higher DMY and higher and lower Cou. Analysis of molecular sequence variation identified seven haplotypes of the trnL-F spacer and 13 haplotypes of the ITS region. Based on haplotype and sequence analyses, the genetic variation of M. officinalis was higher than that of M. albus. Additionally, ITS sequence analysis showed that the variation among accessions was larger than that among species across three geographical areas: Asia, Europe, and North America. Similarly, variation among accessions for both the trnL-F and ITS sequences were larger than the differences between the geographical areas. Our results indicate that there has been considerable gene flow between the two Melilotus species. Our characterization of Cou and the morphological and genetic variations of these two Melilotus species may provide useful insights into germplasm improvement to enhance DMY and Cou.

Analysis of microRNA reveals cleistogamous and chasmogamous floret divergence in dimorphic plant.

Cleistogenes songorica, a grass species that exhibits two spatially different type of inflorescence, chastogamy (CH), flowers localized at the top, and cleistogamy (CL) flowers embedded in leaf sheath. This study aimed at dissecting reasons underlying these distinct floral development patterns at morphological and microRNA level. Phenotyping for CH and CL was conducted and four small RNA libraries were constructed from the CH and CL flowers for high-throughput sequencing to identify the differentiated miRNAs. As results, spikelet, stigma, anther, lemma and lodicule length of CH flowers were found larger than that of CL, and so was seed setting. Also, 17 flower-related differential expression miRNAs were identified which were associated with floral organ development and morphogenesis, and the flower development. Further results showed that miR159a.1-CL3996.Contig2 pair was related to anther development, miR156a-5p-CL1954.Contig2 was linked to response to high light intensity, miR408-3p/miR408d-Unigene429 was related to pollination and Unigene429 positively regulated flower development. To our knowledge, this is the first study on differential miRNA accumulation between CH and CL flowers and our study serves as a foundation to the future elucidation of regulatory mechanisms of miRNAs in the divergent development of CL and CH flowers in a single plant.

EFFICACY AND SAFETY OF BAN HUANG ORAL LIQUID FOR TREATING BOVINE RESPIRATORY DISEASES.

BACKGROUND: Ban Huang oral liquid was developed as a veterinary compound preparation by the Lanzhou Institute of Husbandry and Pharmaceutical Sciences of the Chinese Academy of Agricultural Sciences (CAAS). The purpose of this study was to determine whether the oral liquid preparation of traditional Chinese medicine, Ban Huang, is safe and effective for treating respiratory diseases in cattle. MATERIALS AND METHODS: Acute oral toxicity experiments were conducted in Wistar rats and Kunming mice via oral administration. The minimum inhibitory concentration of the drug against Mycoplasma bovis in vitro with the double dilution method was 500 mg/mL, indicating good sensitivity. The results of laboratory pathogen testing, analysis of clinical symptoms, and analysis of pathological anatomy were combined to diagnose bovine respiratory diseases in 147 Simmental cattle caused by mixed infections of M. bovis, bovine respiratory syncytial virus, bovine parainfluenza virus type 3, and Mannheimia haemolytica. These cattle were randomly divided into three groups: drug treatment group 1 (treated via Tilmicosin injection), drug treatment group 2 (treated with Shuang Huang Lian oral liquid combined with Tilmicosin injection), and drug treatment group 3 (treated with Ban Huang oral liquid combined with Tilmicosin injection). Treatment effects were observed within 7 days. RESULTS: The results showed no toxicity and a maximum tolerated dose greater than 20 g/kg BW. For the 87 cattle in drug-treatment group, the cure rate was 90.80%, whereas the response rate was 94.25%. The cure rate of drug treatment group was increased by 14.13% in comparison with that of drug control group 1 and by 7.47% in comparison with that of drug control group 2 (both P < 0.05). CONCLUSION: This study demonstrates that Ban Huang oral liquid is a safe and effective treatment for bovine respiratory diseases, especially for mixed infection caused by M. bovis, bacteria, and viruses.

The toxicity and the acaricidal mechanism against Psoroptes cuniculi of the methanol extract of Adonis coerulea Maxim.

SCOPE: Adonis coerulea Maxim. is a perennial herbaceous plant that grows in scrub, grassy slope areas, and as traditional medicine it has been used to treat animal acariasis for thousands of years. In this paper, we aimed to study the acute toxicity and cytotoxicity of the methanol extract of A. coerulea (MEAC) in vivo and in vitro for supporting the clinic uses. The acaricidal activity and the mechanism of action against Psoroptes cuniculi were investigated. RESULTS: The results showed that isoorientin, luteolin and apigenin were the primary compounds in MEAC. The toxicity test showed that median lethal dose (LD50) and the 50% inhibitory concentration (IC50) of MEAC were estimated to be more than 5000mg/kg in mice in vivo and more than 50mg/ml against RAW 264.7 and GM00637 cells in the 3-(4, 5-dimethylthiazol-2- yl)-2,5-diphenyltetrazolium bromide (MTT) test. After culturing with MEAC, the activities of superoxide dismutase (SOD), catalase (CAT), malonyldialdehyde (MDA), glutathione-S-transferase (GST), acetylcholinesterase (AChE) and Na+-K+-ATPase of mites were evaluated. Compared with the control group, SOD activity of MEAC-treated group of mites was inhibited, and CAT activity was activated at the preliminary phase but was gradually inhibited over the period of incubation. MDA content reached a peak at 6h and then gradually decreased. However, GST activity in the mites was activated in a dose- and time-dependent manner. AChE and Na+-K+-ATPase activities related to neural conduction, vital functions and the transmembrane ion gradient of the mites were inhibited. CONCLUSION: MEAC is safe in the given doses in both the in vitro and the in vivo tests, can be applied in the clinic and it had good acaricidal activity. The extension of the incubation time in the mites led to dynamic disequilibrium between the production and clearing of superoxide anions, a disruption of the energy metabolism and the transmembrane ion gradient, and the inhibition of motor function. These factors may have resulted in mite death.

Acaricidal activities of the essential oil from Rhododendron nivale Hook. f. and its main compund, δ-cadinene against Psoroptes cuniculi.

In this paper, the acaricidal activities of Rhododendron nivale Hook. f. and its main compound, δ-cadinene were investigated, and the chemical composition of the essential oil was analyzed. The results showed that among aqueous, 70% ethanols, acetic ether, chloroform, petroleum ether and essential oil extracts from the shoots and leaves, the essential oil showed the best in vitro acaricidal activity against adult P. cuniculi, which occurred in a concentration- and time-dependent manner. The median lethal time (LT50) values of four concentrations (33.33-4.17mg/ml) of the essential oil ranged from 1.476 to 25.900h, respectively. After the treatment of P. cuniculi with the essential oil and ivermectin, infected rabbits were free of scabs or secretions in the ear canal by day 20. Then, the percent yield of essential oil from the leaves and shoots was 2.45% (w/w), which includes 50 compounds. The primary component identified was terpenes, and among of compounds identified from the essential oil of R. nivale the highest relative content was δ-cadinene, which also presented the marked acaricidal activity against Psoroptes cuniculi in vitro. These findings provide evidence for the use of acaricides as a traditional medicine and indicate that the essential oil and δ-cadinene could be used to control mites in livestock.

Gymnadenia conopsea (L.) R. Br.: A Systemic Review of the Ethnobotany, Phytochemistry, and Pharmacology of an Important Asian Folk Medicine.

Gymnadenia conopsea (L.) R. Br. (Orchidaceae) is a perennial herbaceous orchid plant that grows widely throughout Europe and in temperate and subtropical zones of Asia. In China, its tuber has been used in traditional Chinese medicines, Tibetan medicines, Mongolian medicines and other ethnic medicines, and taken to treat numerous health conditions. The present paper provides a review of the traditional uses, phytochemistry, biological activities, and toxicology to highlight the future prospects of the plant. More than 120 chemical compounds have been isolated, and the primary components are glucosides, dihydrostilbenes, phenanthrenes, aromatic compounds, and other compounds. G. conopsea and its active constituents possess broad pharmacological properties, such as the tonifying effect, anti-oxidative activity, anti-viral activity, immunoregulatory, antianaphylaxis, antigastric ulcer, sedative, and hypnotic activities, etc. However, overexploitation combined with the habitat destruction has resulted in the rapid decrease of the resources of this plant, and the sustainable use of G. conopsea is necessary to study. Meanwhile, the toxicity of this plant had not been comprehensively studied, and the active constituents and the mechanisms of action of the tuber were still unclear. Further, studies on G. conopsea should lead to the development of scientific quality control and new drugs and therapies for various diseases; thus, its use and development require additional investigation.