Combination Of Essential Oils Research Articles

Effects of dietary thyme essential oil and prebiotic administration on rainbow trout (Oncorhynchus mykiss) welfare and performance

The aim of this study was to examine the combined effects of thyme essential oil (TEO) and prebiotic (Immunogen ®) on growth performance, hematological parameters, innate immunity, and oxidative status of rainbow trout, Oncorhynchus mykiss. For this purpose, the fish (11.92 ± 0.06 g) were fed (3% of biomass) experimental diets that contained 0 (Control; without Immunogen and/or TEO); 1% TEO + 0.1% Immunogen (T1); 1% TEO + 0.2% Immunogen (T2); 2% TEO + 0.1% Immunogen (T3) and 2% TEO + 0.2% Immunogen (T4) for 60 days. According to results, all experimental treatments exhibited similar final weight, weight gain, specific growth rate (SGR), and survival rate (SR), which were significantly higher than those of the control treatment. Although all treatments reduced the feed conversion ratio (FCR) compared to the control group, the lowest value was observed in T4. All experimental treatments showed a significant increase in amylase and protease activity compared to the control group. Moreover, the fish fed on T4 and T1 diets showed the highest and lowest lipase activity. Dietary TEO and Immunogen supplementations significantly increased WBC count and Hb level compared to the control group. Fish fed on control and T2 diets displayed lower hematocrit than fish fed on other experimental diets. The highest and lowest MCH index were recorded in T3 and control groups, respectively. The fish fed diets supplemented with T3 diet presented significantly higher MCV index compared to the control and T2 treatment. All experimental treatments exhibited similar AST, and ALP activities, which were significantly lower than those of the control group. Also, the lowest ALT activity was observed in T2 and T4 treatments compared to other groups. Dietary TEO and Immunogen supplementations significantly enhanced skin mucus total Ig, total protein level, and ACH50, protease, and lysozyme. All experimental treatments exhibited enhanced intestine total Ig, ACH50, and lysozyme level. Dietary thyme essential oil and Immunogen supplementations significantly enhanced liver antioxidant parameters including catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx) and superoxide dismutase (SOD) and decreased MDA production compared to fish fed on control diet. As a result, it can be suggested that the combination of thyme essential oil and Immunogen (specially 2% TEO + 0.2% Immunogen) is useful for enhancing the yield and well-being of farmed rainbow trout.

The impact of lemongrass, oregano, and thyme essential oils on Candida albicans’ virulence factors

Increased systemic infections and growing resistance of Candida species in immunosuppressed people have prompted research for additional treatment options. The purpose of this study was to investigate the potential of lemongrass, oregano, and thyme essential oils tested individually, combined, and combined with the antifungal agents fluconazole and caspofungin to kill Candida albicans isolates in a controlled laboratory setting. A quantitative design was used to analyze the experimental data collected and investigate risk factors related to age, gender, race, and co-morbidities. Kill rates of lemongrass, oregano, and thyme essential oils individually and combined, kill rates of fluconazole, caspofungin, and the kill rates when the antifungals were each combined with the three essential oils were compared using 117 isolates recovered from bloodstream infections between January 2009 through August 1, 2017. There were statistically significant increases in kill rates when the isolates were exposed to any of the combinations of essential oils tested. Data on the covariates age, gender, race, and co-morbidities were assessed for risk factors related to Candida albicans bloodstream infections. The age group 25-34, kidney failure, and solid organ tumor cancer, were all statistically significantly associated with an increased risk for Candida albicans bloodstream infections and multiple organ failure was negatively associated with the risk. Through the use of essential oils, the ability to reduce the number of patients becoming infected with life-threatening yeast infections could not only reduce mortality but also reduce the costs associated with serious infections.

Essential Oil Blends: The Potential of Combined Use for Respiratory Tract Infections.

This study investigated the potential efficacy of 369 commercial essential oil combinations for antimicrobial, anti-toxic and anti-inflammatory activity with the aim of identifying synergy among essential oils commonly used in combination by aromatherapists for respiratory purposes. Essential oil combinations were assessed for their antimicrobial activities using a panel of Gram-positive, Gram-negative, and yeast strains associated with respiratory tract infections. The antimicrobial activity was measured by determining the minimal inhibitory concentration (MIC) of microbial growth. The fractional inhibitory concentration index (ΣFIC) was calculated to determine the antimicrobial interactions between the essential oils in the combination. The toxicity of the essential oil combinations was tested in vitro using the brine shrimp lethality assay, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay on RAW 264.7 mouse macrophage cells and A549 lung cancer cell lines. In addition, an inflammatory response was evaluated measuring nitric oxide production. The essential oils, when in combination, demonstrated an increased antimicrobial effect, a reduction in toxicity and provided improved anti-inflammatory outcomes. Five distinct combinations [Cupressus sempervirens (cypress) in combination with Melaleuca alternifolia (tea tree), Hyssopus officinalis (hyssop) in combination with Rosmarinus officinalis (rosemary), Origanum marjorana (marjoram) in combination with M. alternifolia, Myrtus communis (myrtle) in combination with M. alternifolia and Origanum vulgare (origanum) in combination with M. alternifolia] were found to be the most promising, demonstrating antimicrobial activity, reduced cytotoxicity and improved anti-inflammatory effects. With the increased prevalence of respiratory tract infections and the growing antimicrobial resistance development associated with antimicrobial treatments, this study provides a promising complementary alternative for the appropriate use of a selection of essential oil combinations for use in the respiratory tract.

Influence of Some Antibiotics and Essential Oils Used Alone or in Combination on the Vitality of Presumptive Probiotic Lactic acid Bacteria

Aims: The aim of this study was to assess the in vitro antibacterial activity of selected antibiotics and essential oils alone or in combination, on selected presumptive probiotic lactic acid bacteria.
 Study Design: Experimental studies.
 Place and Duration of Study: Department of Microbiology of the University of Yaounde I between August 2017 and December 2017 (5 months).
 Methodology: The chemical composition of five essential oils was determined by gas chromatography coupled with Solid-phase micro extraction. Then the sensitivity of four lactic acid bacteria to the essential oils and four antibiotics was assessed by the well diffusion and macrodilution method. Subsequently, two essential oils active on these bacteria and broad spectrum antibiotics were combined according to the central composite design plan.
 Results: In general, the chemical composition of essential oils is very diverse, with the example of carvacrol found only in Origanum compactum at 53.24% and thymol in Thymus vulgaris at 56.19% and in Origanum compactum at 15.28%. The antibacterial activity shows that the majority of antibiotics used are active on the bacteria in the study compared to the essential oils where two were active (Origanum compactum and Cymbopogon winterianus). The evaluation of the combinations of essential oils and antibiotics in terms of kinetics has given us three cases: the first case is the one with no acidity or no growth at all; the second is the one where growth is normal; the third where growth is delayed with a more pronounced latency phase.
 Conclusion: This study suggest that the effect of essential oils and medicinal plant used alone or in combination to antibiotics on the gut microbiota have to be evaluated for validation as well as their toxicity activities before using them for human therapy.

Synergism between essential oils: A promising alternative to control Sitophilus zeamais (Coleoptera: Curculionidae)

Combinations of essential oils (EOs) are promising alternatives for pest management in storage grains. A high diversity of compounds in EOs and EO mixtures can increase their toxicity. Additionally, EOs can be useful in insecticide resistance programs. For this reason, we evaluated the fumigant toxicity and repellent effects of six essential oils (Dysphania ambrosioides, DA; Pelargonium graveolens, PE; Piper arboreum, PA; Piper diospyrifolium, PD; Piper gaudichaudianum, PG; and Piper tuberculatum, PT) separately and in binary combinations against adults of Sitophilus zeamais (the maize weevil). First, the insecticidal activity of all possible combinations of the six EOs were assessed. Next, the concentrations needed to cause 50% and 90% mortality (LC50 and LC90, respectively) in the S. zeamais population were determined for the most toxic EOs and mixtures. After that, the synergistic effect of the mixtures was also verified. Finally, the most toxic treatments were assessed to determine the time required to cause mortality of 50% of the population and check the repellent effects on S. zeamais. The binary mixtures of DA and PE, PT, PG, PD, and PA were the most toxic against S. zeamais. The LC50 and LC90 values for these treatments ranged between 1.77 and 2.25 μL L−1 and from 2.42 to 7.64 μL L−1, respectively. The mixtures were more toxic than the EOs tested separately, which indicates the occurrence of synergistic effects. All treatments caused mortality of 50% of the population of S. zeamais in less than 42 h of exposure. In addition, the combinations DA + PE, DA + PT, DA + PG, and DA + PA were repellent to S. zeamais. Based on these results, we conclude that the mixtures of EOs evaluated in this study are highly promising for the control of S. zeamais populations and can also be used for resistance management of this species.

Development of antimicrobial formulation based on essential oils and gamma irradiation to increase the shelf life of boneless chicken thighs

The present study illustrates the development of essential oils (EOs) formulation, and boneless chicken thigh samples were treated with the developed EOs formulation and gamma irradiation to increase the shelf life of treated chicken samples. The binary, tertiary, and quaternary combinations of EOs were developed by the broth dilution method and the fractional inhibitory concentration index. The tertiary combinations of EOs, Thyme + Cannelle + Oregano, showed the best result and exhibited additive (AD) effect against S. enterica, S aureus, and E. coli. The shelf life of the chicken sample increased by 3 days and 8 days when treated by Thyme + Cannelle + Oregano EOs and γ-irradiation, respectively. However, when chicken samples were treated with the combinations of EOs formulations and γ-irradiation, the shelf life of the chicken sample increased by 14 days. The results showed that the combination of EOs treatment with a low dose of γ-irradiation could be used to increase the shelf life of treated food.

Evaluating the effects of high-oil rapeseed cake or natural additives on methane emissions and performance of dairy cows

We evaluated the potential of feeding high-oil rapeseed cake or natural additives as rumen modifiers on enteric methane (CH4) emissions, nutrient utilization, performance, and milk fatty acid (FA) profile of dairy cows. Eight Nordic Red dairy cows averaging (mean ± SD) 81 ± 21 d in milk and 41.0 ± 1.9 kg of milk yield at the beginning of the study were randomly assigned to a replicated 4 × 4 Latin square design with 21-d periods. Treatments comprised grass silage-based diets (45:55 forage to concentrate ratio on dry matter basis) including (1) control containing 19.3% rapeseed meal (CON), (2) CON with full replacement of rapeseed meal with rapeseed cake (RSC), (3) supplementation of CON with 50 g/d of yeast hydrolysate product plus coniferous resin acid-based compound (YHR), and (4) supplementation of CON with 20 g/d of combination of garlic-citrus extract and essential oils in a pellet (GCE). Apparent total-tract digestibility was measured using total collection of feces, and CH4 emissions were measured in respiratory chambers on 4 consecutive days. Data collected during d 17 and 21 in each period were used for ANOVA analysis using a mixed model. Treatments did not affect dry matter intake (DMI), whereas feeding RSC increased crude protein and ether extract digestibility compared with the other diets. Emissions of CH4 per day, per kilogram of DMI, and per kilogram of energy-corrected milk, and gross energy intake were lower for RSC compared with other diets. We found no effect of YHR on daily CH4 emissions, whereas CH4 yield (g of CH4/kg of DMI or as percentage of gross energy intake) decreased with GCE compared with CON. Treatments did not influence energy balance. Further, RSC reduced the proportion of N intake excreted in feces, and YHR improved N balance compared with CON diet. Feeding RSC resulted in greatest yields of milk and energy-corrected milk, and feed efficiency. Relative to the CON diet, RSC decreased saturated FA by 10% in milk fat by increasing cis-monounsaturated FA but also increased the proportion of trans FA. Proportion of odd- and branched-chain FA increased with GCE and YHR compared with CON. We conclude that replacing rapeseed meal by rapeseed cake decreased CH4 emissions, whereas YHR or GCE had no effect on CH4 emissions in this study.

Synergy between Pelargonium endlicherianum essential oil and conventional antibiotics against Neisseria meningitidis and Haemophilus influenzae

This study investigated the bactericidal effects of the essential oil of Pelargonium endlicherianum in combination with four antibiotics commonly used in the treatment of bacterial meningitis (penicillin, ampicillin, ciprofloxacin, and gentamicin) against the meningitis causative pathogens Neisseria meningitidis and Haemophilus influenzae. The phagocytic effects of these combinations were also tested against human leukocyte cells. The bactericidal effect of P. endlicherianum essential oil (PEO) and antibiotic combinations was dynamically detected by time-kill assay. The function of PEO and antibiotic in permeating outer membrane barriers, when used singly or in combination, was analyzed by UV spectrophotometer.The interactions between antibiotic and essential oil were calculated according to the fractional inhibitory concentration (FIC) index. While a synergistic effect of the ciprofloxacin + PEO combination was determined on N. meningitidis (FIC≤ 0.5), an additive effect was observed on H. influenzae (FIC= 1). Combined use of PEO with gentamicin showed a synergistic effect against N. meningitidis and H. influenzae (FIC ≤ 0.5). The antimicrobial effect of the penicillin + PEO combination was higher than that of penicillin + PEO used alone. The ampicillin + PEO combination had a synergistic effect on N. meningitidis and an additive effect on H. influenzae. The results of our study showed that the essential oil increases membrane permeability activity and also has phagocytic activity in human leukocyte cells. Combining antibiotics with essential oils that target resistant bacteria may open up new options in combating microbial resistance.

Repellent activity of three essential oils against dengue and filarial vector mosquitoes

Cymbopogon nardus, Syzygium aromaticum, and Citrus sinensis have been well known to possess insecticidal activity. Here, we investigated the repellent activity of individual of three essential oils (C. nardus, S. aromaticum, C. sinensis) and a combination of essential oils (C. nardus + C. sinensis, C. nardus + S. aromaticum, and C. sinensis + S. aromaticum) against a dengue vector, Aedes aegypti and a filarial vector, Culex quinquefasciatus. The essential oils (1 mg) were smeared evenly on the surface of the left arm (three volunteers) and then subjected to 25 females of A. aegypti and C. quinquefasciatus in the cage. The number of landed mosquitos was observed every hour for 7 hours to determine the protection index (PI). The result showed that individuals of three essential oils and a combination of essential oils generated > 90% of PI until 7 hours of application for both mosquitos species tested. Overall, the data demonstrated that individual of three essential oils, i.e., C. nardus, S. aromaticum, C. sinensis, and various combinations of essential oils (C. nardus + C. sinensis; C. nardus + S. aromaticum; C. sinensis + S. aromaticum) are promising as repellent agents against A. aegypti and C. quinquefasciatus.

ИЗМЕНЕНИЕ ФАКТОРОВ НЕСПЕЦИФИЧЕСКОЙ ЗАЩИТЫ РОТОВОЙ ПОЛОСТИ У ДЕТЕЙ С ГИНГИВИТОМ НА ФОНЕ ПРИМЕНЕНИЯ ЭФИРНЫХ МАСЕЛ И БЕНТОНИТОВОЙ ГЛИНЫ

Introduction. The prevention and treatment of inflammatory periodontal disease in children remains a topical issue in dentistry. This is due to the increasing prevalence of this disease in the early age groups. Remission periods of diseases in children are short. Therefore, treatment and preventive measures of periodontal tissue diseases in this population should be primarily focused on the regulation of nonspecific protection system at the local level. The components of the system are secretory immunoglobulin A, lysozyme and modified albumin. Using means of natural origin as regulators will allow to optimize treatment regimens and reduce allergic reactions. Aim. The aim of the study was to study the effectiveness of the complex for applications to the gingiva, consisting of means of natural and mineral origin of the Crimean deposit, in the regulation of local oral immunity in children with chronic catarrhal gingivitis. Material and methods. Twenty-eight children aged 11–12 years with moderate chronic catarrhal gingivitis were examined. The control group consisted of 12 healthy children of the same age. We used applications of phytopreparations on the gingiva in the treatment of chronic catarrhal gingivitis daily for 20 minutes, No 10. Complex for applications was essential oils of peppermint, pine, fennel, eucalyptus (1:1:1:1) on the basis of bentonite clay (10:1). Clinical and laboratory parameters were determined in the dynamics. Statistical processing was performed using the standard method of variation statistics. Comparison of the mean indices was conducted using Student’s t-criterion. Differences were considered reliable at p<0,05. Results and discussion. The criterion for the effectiveness of treatment was the positive dynamics of paraclinical indices (p<0,001). Indicators of oral dysbiosis decreased and reached values of oral microbiocenosis of healthy children. The content of sIgA increased 1,8 times (p<0,001). The level of salivary lysozyme activity increased 1,7 times (p<0,001) and reached normal values. The concentration of modified albumin decreased 1,6-fold. Conclusion. Inclusion of applications of the combination of essential oils based on bentonite clay of the Crimean deposit into the plan of treatment of chronic catarrhal gingivitis in children contributes to a pronounced relief of the signs of inflammation in the gingiva. The proposed complex has a pathogenetic orientation in the treatment of chronic catarrhal gingivitis, which is confirmed by the pronounced dynamics of nonspecific immunity parameters.

Effect of emulsifier and droplet size on the antibacterial properties of emulsions and emulsion‐based films containing essential oil compounds

In this study, the physical properties and antibacterial activity of emulsions prepared through four different combinations of essential oils (EOs) (trans-cinnamaldehyde [TC] and citral) and emulsifiers (sodium caseinate and lecithin) were investigated. The results showed that the four different nanoemulsions had nanometric sizes ranging from 142.3 to 273.1 nm, exhibiting high negative zeta potential (−25.3 to −42.7 mV) at pH 7. Nanoemulsions containing TC exhibited higher antimicrobial activities against Escherichia coli and Listeria innocua than nanoemulsions containing citral irrespective of the emulsifier used. These nanoemulsions were then incorporated into low methoxyl pectin films and the obtained results showed that films containing TC stabilized by sodium caseinate had a larger inhibition zone than the one stabilized by lecithin irrespective of the EO used. Finally, the obtained results showed that nanosized emulsions were more stable than microsized ones, but no significant differences were observed concerning their antibacterial activity. Practical applications This work should be an interesting basis for developing innovative antimicrobial packaging containing nanoemulsions. The interfacial membrane that surrounds the droplets of the emulsified antimicrobial provides an additional barrier that the active agent should cross before leaving the polymer matrix of the packaging film. Thus, the release kinetics will be modified and the efficiency could be improved.

The effect of essential oils on cholesterol content in chicken meat

The study aimed to investigate cholesterol content in chicken breast and thigh muscles by the influence of feed supplements of various content of essential oils. The experiment was carried out under practical conditions in a poultry farm with broiler chickens of the Cobb 500 hybrid combination according to the feed supplement used thyme essential oil, cinnamon essential oil, commercial citrus fruit essential oil, and their combination. The control group was without the use of experimental feed supplements and commercial coccidiostats were used in their feed mixtures. The experiment lasted 40 days in welfare conditions. Broiler chickens were used for sample preparation of breast and thigh muscles with the skin and their analysis for dry matter, fat and cholesterol contents. Samples were analyzed using a Nicolet 6700 FT-IR Fourier transform infrared spectrometer. The results were processed by the SAS system program, version 8.2. The results, which were evaluated, indicated a tendency to reduce the cholesterol content in chicken breast muscle due to cinnamon essential oil and the combination of cinnamon essential oil with citrus fruit essential oil as well as thyme essential oil with citrus fruit essential. In the achieved results of dry matter, fat, and cholesterol content in breast and thigh muscles, the difference between the effects of the used feed supplements based on essential oils and concerning the control group were not statistically significant p >0.05. The correlation was a statistically significant strong linear relation only between dry matter content and fat content due to thyme and cinnamon essential oils. In conclusion, it was stated that the investigation of the feed supplement effect based on essential oils is an open question concerning the production of safe food of animal origin.

Antifungal and synergistic activities of some selected essential oils on the growth of significant indoor fungi of the genus Aspergillus

The study aimed to assess the antifungal activity of twenty–five essential oils (EOs) and the potential synergistic activity of the most effective EOs against significant indoor fungi of the genus Aspergillus [A. fumigatus (KBio–122), A. flavus (KBio–134), A. terreus (KBio–145) and A. niger (KBio–202)]. The chemical composition of all EOs was evaluated by the gas chromatography coupled with mass spectrometry (GC/MS) and gas chromatography with flame ionization detector (GC–FID) analysis. The antifungal susceptibility of EOs was evaluated by using the broth microdilution method. The most effective EOs were selected to determine the minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations (MFCs) at a concentration range from 256 to 0.125 μg/mL. For the synergistic activities, the most effective EOs were tested using the chessboard pattern. The most sensitive strain to treatments with essential oils alone and in the combination of EOs was A. flavus (KBio–134). The chessboard assay showed that combinations of lemongrass and thyme EOs proved the most potent synergistic antifungal activity (FICI = 0.1875) against A. fumigatus (KBio–122). The synergy displayed by a combination of some EOs may be used to control fungal growth or increasing resistance to available synthetic antifungals, consequently permitting the reduction of their most active doses.

299 Combination of Essential Oils, α-amylase and 25-hydroxy-vitamin-d3 Increases Carcass Production by feedlot Cattle

Abstract Our objective was to evaluate the effects of associating feed additives on feedlot performance of Nellore bulls. One hundred and sixtieth Nellore bulls (iBW, 380± 5.8 kg; age, 18 mo) were allocated in 20 pens (n = 8/pen), in a completely randomized block design, according to the treatments: 1) MON+VM - Sodium Monensin (MON, 25 mg/kg DM, Elanco Animal Health, Indianapolis, IN), + Virginiamycin (VM, 30 mg/kg DM, Phibro Animal Health Corporation, Guarulhos, São Paulo), and 2) CRD - Crina® RumistarTM + HyD® - a blend of essential oils, 90 mg/kg DM + exogenous α-amylase, 560 mg/kg DM + 25-hydroxy-vitamin-D3 at 1 mg/animal/d (DSM Nutritional Products, Basel, Switzerland). The initial BW was utilized as a criterion for block formation. Pens were considered experimental units (10 pens per treatment). Animals were adapted to the diets during 14 days with gradually increasing concentrate level from 70% to 86% of diet DM, and fed for 90 days. Weight assessments were performed at day 0 and 90 after 14h fasting. Data were analyzed using PROC MIXED of SAS and means were separated using the PDIFF statement, adopting P < 0.05. Feeding CRD increased the DMI (12.13 kg vs. 10.52 kg, P < 0.01), ADG (1.86 vs. 1.63 kg/d, P < 0.01) and final BW (542 vs. 521 kg, P < 0.01) compared to animals fed MON+VM. In addition, feeding CRD increased final HCW in 15 kg (311 vs. 296 kg; P < 0.01), carcass ADG in 0,16 kg/d (1.31 vs. 1.15 kg/d; P < 0.01) and dressing percentage in 0.48 percent points (57.34 vs. 56.86%; P < 0.01) compared to MON+VM. The G:F (0.155, P = 0.53) and biological efficiency (137.9 kg DM/15 kg carcass, P = 0.87) were similar among treatments. Therefore, the inclusion of Crina® RumistarTM +HyD® can be used as a tool to increase carcass gain of feedlot finishing cattle.

Preliminary Study on Citrus Oils Antibacterial Activity Measured by Flow Cytometry: A Step-by-Step Development.

Essential oils of Citrus sinensis and Citrus latifolia have shown biological functions as antiseptics, anti-inflammatories, antioxidants, antifungal and antimutagenic, so the evaluation of their antibacterial capacity, by themselves or in combination with standard antibiotics, presents an alternative for infection treatment. Flow cytometry opens the door for the design of faster and more accurate measurement of antibacterial activity. We use a SYTO9/PI staining system on E. coli ATCC 25922 to determine antibacterial activity by counting live and dead cells through flow cytometry. We found that dual staining showed highly variable results due to wavelength overlapping and instead we used fluorochrome individual staining that highly correlated with viable counts. Chloramphenicol and cefotaxime treatments did not present a dose-response behavior, rendered diffuse readings and/or gave filament formation on fluorescence microscopy. Amikacin was a better comparison standard because it presented a dose-response behavior. Essential oils had low antibacterial activity as compared to amikacin, with a maximum of 10% and 20% for C. latifolia and C. sinensis, respectively. Combinations of essential oils with antibiotic resulted in an unforeseen strong inhibition of amikacin activity. Although a low antibacterial activity was found, a series of standardization steps are proposed for antibacterial activity measurement by flow cytometry.

Karakterisasi Obat Nyamuk Bakar Herbal Dengan Bahan Baku Limbah Gagang Cengkeh dan Tempurung Biji Pala

The increase of of cloves and sed nutmeg productions effects in its waste into the environment. However, the cloves and nutmeg shell could be use as mosquito repellent. The combination of essential oil extracted from cloves and nutmeg shell functions well as repellent for mosquitos. This research aims to identified the characteristics of essential oil extracted from these two spices by analysing its water, carbon aand ash concentrate. Therefore, we found that water level in U1 was 1.4573, and U2 was 1.6070, hence ash rate for U1 was  7,8674, U2 was 7,49224 and for carbon it was U1 24,4496 U2 23,5778. At the end we could say that for and ash it has already meet the SNI standard, while carbon hasn’t meet SNI standard yet.  Key words: Herbal mosquito repellent; waste; nutmeg shell waste.

THE EFFECT OF BACTERIOCIN ENTEROCOCCUS ITALICUS ONU 547 AND ESSENTIAL OILS ON THE GROWTH OF OPPORTUNISTIC MICROORGANISMS

Purpose. To evaluate the effectiveness of bacteriocin of Enterococcus italicusONU 547 and essential oils individually and in combination against indicatorstrains of opportunistic pathogens. Methods. The antagonistic activity of essential oils and bacteriocin was determined by the optical density of the bacterial suspension of test strains (Salmonella enterica NCTC 6017, Escherichia coli АТСС 25922, Pseudomonas aeruginosa АТСС 27853, Pseudomonas putida KT 2440, Enterococcus faecalis АТСС 29212, Klebsiella pneumoniae АТСС 10031,Staphylococcus aureus АТСС 25923). The index of fractional inhibitory activity(FIC) was calculated by adding the FIC values of bacteriocin E. italicus ONU 547and the studied essential oils. The results. The maximum antimicrobial effect was caused by the essential oils of Anisum officinalis and Melissa officinalis, which suppressed the growth of test strains more than 50.0%. When using bacteriocin of E. italicus ONU 547 suppression of viability of indicator strains did not exceed 30.0%. Experimental combinations of essential oils and bacteriocin caused 4 types of consequences: synergy effect; additive effect; antagonistic effect; interactiveeffect (no interaction effect). In most cases, the combined action of bacteriocinand essential oils caused an additive effect. Conclusions. This study demonstrated the potential of using different combinations of natural antimicrobial compounds. The obtained results provide grounds for further development and optimization ofcombinations of essential oils and probiotic strains of microorganisms for use in food and pharmaceutical biotechnology.

Minimum inhibitory concentrations of commercial essential oils against common chicken pathogenic bacteria and their relationship with antibiotic resistance.

AimsWe investigated the antibacterial effect of seven essential oils (EOs) and one EO‐containing liquid phytogenic solution marketed for poultry and pigs (‘Product A’) on chicken pathogens, as well as the relationship between minimum inhibitory concentration (MIC) in EOs and antibiotics commonly administered to chicken flocks in the Mekong Delta (Vietnam).Methods and ResultsMicellar extracts from oregano (Origanum vulgare), cajeput (Melaleuca leucadendra), garlic (Allium sativum), black pepper (Piper nigrum), peppermint (Mentha × piperita L.), tea tree (Melaleuca alternifolia), cinnamon (Cinnamomum zeylanicum) EOs and Product A were investigated for their MIC against Avibacterium endocarditidis (N = 10), Pasteurella multocida (N = 7), Ornitobacterium rhinotracheale (ORT) (N = 10), Escherichia coli (N = 10) and Gallibacterium anatis (N = 10). Cinnamon EO had the lowest median MIC across strains (median 0.5 mg/ml [IQR, interquartile range 0.3–2.0 mg/ml]), followed by Product A (3.8 mg/ml [1.9–3.8 mg/ml]), oregano EO (30.4 mg/ml [7.6–60.8 mg/ml]) and garlic 63.1 mg/ml [3.9 to >505.0 mg/ml]. Peppermint, tea tree, cajeput and pepper EOs had all MIC ≥219 mg/ml. In addition, we determined the MIC of the 12 most commonly used antibiotics in chicken flocks in the area. After accounting for pathogen species, we found an independent, statistically significant (p < 0.05) positive correlation between MIC of 10 of 28 (35.7%) pairs of EOs. For 67/96 (69.8%) combinations of EOs and antibiotics, the MICs were correlated. Of all antibiotics, doxycycline was positively associated with the highest number of EOs (peppermint, tea tree, black pepper and cajeput, all p < 0.05). For cinnamon, the MICs were negatively correlated with the MICs of 11/12 antimicrobial tested (all except colistin).ConclusionsIncreases in MIC of antibiotics generally correlates with increased tolerance to EOs. For cinnamon EO, however, the opposite was observed.Significance and Impact of the StudyOur results suggest increased antibacterial effects of EOs on multi‐drug resistant pathogens; cinnamon EO was particularly effective against bacterial poultry pathogens.

New Insight into the Chemical Composition, Antimicrobial and Synergistic Effects of the Moroccan Endemic Thymus atlanticus (Ball) Roussine Essential Oil in Combination with Conventional Antibiotics.

This study reported the volatile profile, the antimicrobial activity and the synergistic potential of essential oil (EO) from the Moroccan endemic Thymus atlanticus (Ball) Roussine, in combination with the antibiotics ciprofloxacin and fluconazole for the first time, to the best of our knowledge. The EO chemical composition was determined by gas chromatography coupled to mass spectrometry (GC-MS) analysis and the antimicrobial activity assessed by the disc diffusion method against three Gram positive (Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus) and three Gram-negative bacteria (Pseudomonas aeruginosa, Escherichia coli and one clinical isolate, Klebsiella pneumonia). The antifungal activity was evaluated in four pathogenic yeasts (Candida albicans, C. glabrata, C. krusei and C. parapsilosis). The minimum inhibition concentration (MIC) and the synergistic effect with ciprofloxacin and fluconazole were determined by the two-fold dilution technique and checkerboard test, respectively. Twenty-one constituents were identified by GC-MS in the EO, including carvacrol (21.62%) and borneol (21.13%) as the major components. The EO exhibited a significant antimicrobial activity with inhibition zones ranging from 0.7 mm to 22 mm for P. aeruginosa and B. subtilis, respectively, and MIC values varying from 0.56 mg/mL to 4.47 mg/mL. The fractional inhibitory concentration index (FICI) values ranged from 0.25 to 0.50 for bacteria and from 0.25 to 0.28 for yeasts. The maximum synergistic effect was observed for K. pneumonia with a 256-fold gain of antibiotic MIC. Our results have suggested that EO from T. atlanticus may be used alone or in association with antibiotics as a new potential alternative to prevent and control the emergence of resistant microbial strains both in the medical field and in the food industry.

Influence of dietary supplementation of clove and rosemary essential oils or their combination on growth performance, immunity status, and blood antioxidant of growing rabbits.

This study aimed to assess the dietary effects of rosemary and clove essential oils separately and in combination on the growth performance; immunological, hematological, and physiological responses; and antioxidant status of growing rabbits. One hundred forty-four of 42-day-old growing V-line rabbits (both sexes with initial live body weights of 765 ± 6g) were randomly allocated into four treatment groups of 36 rabbits each. Each group was further sub-divided into 12 replicates of 3 rabbits in a completely randomized design. The 1st group was fed a basal diet free of additives and served as the control group, the 2nd and 3rd groups were fed basal diets supplemented with rosemary and clove essential oils, respectively, at doses of 400mg/kg diet. The 4th group received a basal diet supplemented with a combination of clove and rosemary essential oils at doses of 200mg/kg diet each. The results showed that the different supplementations did not influence rabbit performance or immunological traits. Opposite to performance or immunological traits, differences in red blood cells and hemoglobin value among all dietary treatments were improved (P < 0.05). Dietary essential oil supplementation with clove, rosemary oil, or a mixed of both increased (P < 0.05) blood concentrations of catalase, superoxide dismutase, and glutathione peroxidase by 47, 42, and 7%; 56, 35, and 36%; and 40, 39, and 37%, respectively, in supplemented rabbits versus control rabbits. In conclusion, clove and/or rosemary essential oils can potentially be used in rabbit diets to improve antioxidant status without change in rabbit's growth performance or immunological parameters.