Basma Najar

Antibacterial and Antifungal Activity of Essential Oils against Pathogens Responsible for Otitis Externa in Dogs and Cats

Background: Essential oils (EOs) are recommended by some veterinarians to treat otitis externa in pets, but data about their efficacy in scientific literature are very scant. Methods: Nine commercial EOs, from roman chamomile (Anthemis nobilis L.), star anise (Illicium verum), lavender (Lavandula hybrida), litsea (Litsea cubeba (Lour.) Pers.), basil (Ocimum basilicum L.), oregano (Origanum vulgare L. subsp. hirticum), rosemary (Rosmarinus officinalis L.), clary sage (Salvia sclarea L.), and thyme (Thymus vulgaris L.) were tested against bacterial and fungal pathogens previously isolated from dogs and cats with otitis externa. In particular, the analyses were carried out against Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus pseudointermedius, Aspergillus niger, Aspergillus fumigatus, Aspergillus terreus, Candida albicans, Candida tropicalis, Trichosporon sp., and Rhodotorula sp. Results: O. vulgare and S. sclarea showed superior antibacterial activity, even if not against all the strains. Trichosporon sp., C. albicans, and A. terreus were insensitive to most Eos, while other yeasts and molds showed different degrees of sensitivity. In particular, most fungi were inhibited by O. vulgare and R. officinalis. Conclusions: The obtained results suggest that some EOs could be included in treatment as an alternative therapeutic option in bacterial otitis complicated by fungi, in association with conventional drugs.

Traditional Mediterranean plants: characterization and use of an essential oils mixture to treat Malassezia otitis externa in atopic dogs.

Abstract Several plants extracts from Mediterranean countries are traditionally employed in skin troubles both in humans and in animals. Malassezia pachydermatis is a lipophylic yeast responsible for otitis externa and dermatitis in dogs and for cutaneous and systemic disease in humans. Five mixtures of essential oils obtained from Mediterranean plants (Citrus paradisi, Salvia sclarea, Ocimum basilicum, Rosmarinus officinalis, Citrus limon, Anthemis nobilis, Lavandula hybrida and Thymus vulgaris) provided with antifungal and/or anti-inflammatory action assayed in vitro, were tested in vivo versus M. pachydermatis to treat once daily for 2 weeks 25 atopic dogs with Malassezia otitis externa. Mixture composed by C. limon 1%, S. sclarea 0,5%, R. officinalis 1%, A. nobilis 0,5% yielded excellent results in all treated dogs. Despite of clinical resolution after all treatments the number of blastospores did not decrease. This study confirms recent findings suggesting a multifactorial alternative approach for the management of canine Malassezia otitis.

Sub-inhibitory stress with essential oil affects enterotoxins production and essential oil susceptibility in Staphylococcus aureus

Abstract Fourteen wild strains of Staphylococcus aureus positive for gene sea were tested for enterotoxins production and the minimum inhibitory concentration of Leptospermum scoparium, Origanum majorana, Origanum vulgare, Satureja montana and Thymus vulgaris essential oils (EOs) were determined. After this trial, bacteria stressed with sub-inhibitory concentration of each EO were tested for enterotoxins production by an immunoenzymatic assay and resistance to the same EO. Oregano oil exhibited the highest antibacterial activity followed by manuka and thyme oils. After the exposure to a sub-inhibitory concentration of EOs, strains displayed an increased sensitivity in more than 95% of the cases. After treatment with oregano and marjoram EOs, few strains showed a modified enterotoxins production, while 43% of the strains were no longer able to produce enterotoxins after treatment with manuka EO. The results obtained in this study highlight that exposure to sub-inhibitory concentration of EO modifies strains enterotoxins production and EOs susceptibility profile.

Chemical Composition and In Vitro Antimicrobial Efficacy of Sixteen Essential Oils against Escherichia coli and Aspergillus fumigatus Isolated from Poultry

Escherichia coli and Aspergillus fumigatus are two pathogens largely present among poultry. They can cause mild or severe forms of disease, and are associated with significant economic losses. The aim of the present study was to investigate the chemical composition and the in vitro antimicrobial activity of sixteen essential oils (EOs) and five mixtures against E. coli and A. fumigatus strains previously isolated from poultry. The study was performed with the following EOs: Aloysia tryphilla, Boswellia sacra, Cinnamomum zeylanicum, Citrus aurantium, Citrus bergamia, Citrus limon, Citrus reticulata, Cymbopogon citratus, Eucalyptus globulus, Lavandula hybrida, Litsea cubeba, Ocimum basilicum, Melaleuca alternifolia, Mentha piperita, Pelargonium graveolens, and Syzygium aromaticum. Moreover, the following mixtures were also tested: L. cubeba and C. citratus (M1), L. cubeba and A. triphylla (M2), A. triphylla and C. citratus (M3), A. triphylla, C. citratus and L. cubeba (M4), S. aromaticum and C. zeylanicum (M5). One hundred and ninety-one compounds were identified in the tested EOs and mixtures. MIC determination found good anti-E. coli activity with C. zeylanicum (2.52 mg/mL), C. citratus (1.118 mg/mL), L. cubeba (1.106 mg/mL), M. piperita (1.14 mg/mL) and S. aromaticum (1.318 mg/mL) EOs. Among the mixtures, M5 showed the best result with a MIC value of 2.578 mg/mL. The best antimycotic activity was showed by A. triphylla (0.855 mg/mL), followed by C. citratus (0.895 mg/mL), while C. aurantium, M. piperita, B. sacra and P. graveolens did not yield any antifungal effect at the highest dilution. The mixtures exhibited no antifungal activity at all. This study shows promising results in order to use EOs in the environment for disinfection purposes in poultry farms and/or in hatcheries.

Stonebrood and chalkbrood in Apis mellifera causing fungi: in vitro sensitivity to some essential oils

Abstract Aim of the present study was to evaluate the in vitro antimycotic activity of 17 chemically defined essential oils (EOs) both alone and as a mixture, against agents responsible for stonebrood caused by Aspergillus flavus, and chalkbrood caused by Ascosphaera apis in European honeybees. Cinnamomum zeylanicum yielded the lowest MIC value against A. flavus, but was not effective against A. apis, while Litsea cubeba and Pelargonium graveolens appeared to be effective against all checked fungi. Aspergillus niger showed the lower sensitivity. Two mixtures composed by L. cubeba, C. zeylanicum and Cymbopogon flexuosus (M1) and by L. cubeba, C. zeylanicum, P. graveolens and C. flexuosus (M2), respectively, were tested, both resulting effective. The components of M1 showed a synergistic effect. The use of mixtures allowed to decrease the total amount of EOs. The use of these products could be of interest for an alternative natural approach in honeybee disease management.

In Vitro Activity of 30 Essential Oils against Bovine Clinical Isolates of Prototheca zopfii and Prototheca blaschkeae

Protothecal mastitis poses an emergent animal health problem in dairy herds, with a high impact on dairy industries, causing heavy economic losses. Current methods of treating protothecal infections are ineffective, and no drug is licensed for use in cattle. The aim of the present study was to check the antialgal activity of 30 chemically defined essential oils (EOs) against Prototheca zopfii and Prototheca blaschkeae isolated from the milk of dairy cows with mastitis. A microdilution test was carried out to estimate the antialgal effectiveness of the selected chemically defined EOs. The microdilution test showed different degrees of inhibition among the examined Prototheca species. The activity of some of the examined EOs seem interesting. In particular, Citrus paradisi yielded the lowest minimal inhibitory concentration values (0.75%) for both algal species. P. zopfii appeared to be more sensitive to EOs in comparison to P. blaschkeae. The present study investigated the in vitro susceptibility of P. zopfii and P. blaschkeae to a wide range of EOs, obtained from different botanical families. Further investigations are necessary to evaluate the efficacy of EO-based formulations intended for the disinfection of both udder and milking products.

Salvia broussonetii Benth.: aroma profile and micromorphological analysis

Abstract The volatile profiles (VOC) and the essential oil (EO) composition from the aerial parts of Salvia broussonetii were analysed. Sesquiterpene hydrocarbons dominate the VOCs from leaves (95.7%) and flowers (67.6%), followed by monoterpene hydrocarbons (2.6 and 29.7%, respectively). The main common compounds are germacrene D, β-bourbonene, α-pinene, α-copaene and α-gurjunene, even if with divergent relative abundances. In the leaf EOs the sesquiterpenes prevail, even if not overwhelmingly (about 50.0%), followed by monoterpenes (23.0–35.0%) and by minor fractions of diterpene hydrocarbons and non-terpene derivates. The most abundant common compounds across the two sampling periods are α-pinene, β-pinene, isobornyl acetate, α-gurjenene, germacrene D and bifloratriene. A morphological characterisation of the trichomes responsible for the productivity in terpenes was also performed. Four different morphotypes were observed on the above ground organs of S. brussonetii: peltates and capitates of type II and III resulted the only producers of volatile substances.

Latitude and Altitude Influence Secondary Metabolite Production in Peripheral Alpine Populations of the Mediterranean Species Lavandula angustifolia Mill.

Lavandula angustifolia Mill. has a great economic importance in perfumery, cosmetics, food manufacturing, aromatherapy, and pharmaceutical industry. This species finds its phytosociological optimum in the sub-Mediterranean region. Latitudinal and altitudinal gradients are expected to affect species diversification in peripheral alpine populations. In this study, phenotypic traits including morphometric parameters, volatile organic compounds (VOCs) and essential oils (EOs) were analyzed in lavender peripheral populations selected in order to explore different ecological conditions. Plants were cultivated under uniform conditions to observe variations due to the genetic adaptation to native environments and to exclude the short-term response to environmental factors. Results showed qualitatively and quantitatively intra-specific variations in secondary metabolites, mainly along the latitudinal gradient, while minor effect was attributable to the altitude. This latter affected more the morphometric parameters. As the latitude augmented, VOCs showed lower content of monoterpene hydrocarbon (mh) and higher content of oxygenated monoterpenes (om); whereas EOs showed higher content of mh and non-terpene derivatives (nt) and lower content of sesquiterpene hydrocarbons (sh). Lavender aroma and EO composition varied in every population, for a total of 88 and 104 compounds identified, respectively. Eleven and 13 compounds were responsible for 95% of the dissimilarity, with linalool, linalyl acetate and 1,8-cineole as major contributors. As the latitude augmented, linalool decreased and 1,8-cineole increased while linalyl acetate content was unaffected. These results are discussed with regards to the potential adoption of the lavender peripheral alpine populations for the improvement of quality and productivity of lavender cultivations, especially in mountainous areas.

Activity of selected essential oils on spoiling fungi cultured from Marzolino cheese

Microscopic fungi can be present on a variety of foodstuff, including cheese. They can be responsible for fungal spoilage, causing sensory changes making food unacceptable for human consumption, and posing severe health concerns. Furthermore, some of these organisms are able to resist antimicrobial preservatives provided for by law. Antifungal activity of 15 chemically defined EOs, alone and in mixture, were checked by a microdilution test against isolates of Penicillium funiculosum and Mucor racemosus cultured from rinds of Marzolino, a typical Italian fresh pecorino cheese. Origanum vulgare yielded the lowest MIC values, followed by Salvia sclarea, Ocimum basilicum and Cymbopogon citratus, while Citrus paradisi and Citrus limon were not active. All mixtures showed antifungal activity at lower concentration with respect to MIC values of each EO component, when not in combination. This study is the first to describe the setting up of EOs mixtures to limit spoiling moulds.

Sub-inhibitory concentration of essential oils induces antibiotic resistance in Staphylococcus aureus

Abstract Fourteen Staphylococcus aureus wild strains were stressed with sub-inhibitory concentration of five essential oils: Leptospermum scoparium (manuka), Origanum majorana (marjoram), Origanum vulgare (oregano), Satureja montana (winter savoury) and Thymus vulgaris (thyme). Antibiotics susceptibility profiles of the strains were determined by agar disk diffusion method before and after EOs treatment. The following antibiotics were employed: amoxicillin-clavulanic acid, amikacin, ampicillin, amoxicillin, aztreonam, ceftazidime, cephalothin, ciprofloxacin, colistin, cefotaxime, doxycycline, enrofloxacin, erythromycin, gentamicin, cephalexin, neomycin, piperacillin, rifampin, streptomycin, trimethoprim-sulphamethoxazole, tetracycline and tobramycin. Before EOs treatment, strains were susceptible to all antibiotics except for aztreonam and colistin. After exposure to sub-inhibitory EOs concentration of manuka, marjoram and oregano, several modifications in antibiotics susceptibility profiles were detected. Conversely, few modifications were induced by winter savoury and thyme EOs. Moreover, occurrence of resistances seems uncorrelated with drug classes as low concentration of EO could induce phenotypic changes in susceptible bacteria leading to antibiotic resistance phenomena.