Preliminary assessment of the mucosal toxicity of tea tree (Melaleuca alternifolia) and rosemary (rosmarinus officinalis) essential oils on novel porcine uterus models

Antimicrobial resistance, an ever-growing global crisis, is strongly linked to the swine production industry. In previous studies, Melaleuca alternifolia and Rosmarinus officinalis essential oils have been evaluated for toxicity on porcine spermatozoa and for antimicrobial capabilities in artificial insemination doses, with the future perspective of their use as antibiotic alternatives. The aim of the present research was to develop and validate in vitro and ex vivo models of porcine uterine mucosa for the evaluation of mucosal toxicity of essential oils. The in vitro model assessed the toxicity of a wider range of concentrations of both essential oils (from 0.2 to 500 mg/mL) on sections of uterine tissue, while the ex vivo model was achieved by filling the uterine horns. The damage induced by the oils was assessed by Evans Blue (EB) permeability assay and histologically. The expression of ZO-1, a protein involved in the composition of tight junctions, was assessed through immunohistochemical and immunofluorescence analysis. The results showed that low concentrations (0.2\u20130.4 mg/mL) of both essential oils, already identified as non-spermicidal but still antimicrobial, did not alter the structure and permeability of the swine uterine mucosa. Overall, these findings strengthen the hypothesis of a safe use of essential oils in inseminating doses of boar to replace antibiotics

Potential Applications of Essential Oils for Environmental Sanitization and Antimicrobial Treatment of Intensive Livestock Infections

The extensive use of antibiotics has contributed to the current antibiotic resistance crisis. Livestock infections of Salmonella spp, Clostridium spp. and E. coli antimicrobial-resistant bacteria represent a public threat to human and animal health. To reduce the incidence of these zoonoses, essential oils (EOs) could be effective antibiotic alternatives. This study aims at identifying EOs safe for use, effective both in complementary therapy and in the environmental sanitization of intensive farming. Natural products were chemo-characterized by gas chromatography. Three S. Typhimurium, three C. perfringens and four E. coli strains isolated from poultry and swine farms were used to assess the antimicrobial properties of nine EOs and a modified GR-OLI (mGR-OLI). The toxicity of the most effective ones (Cinnamomum zeylanicum, Cz; Origanum vulgare, Ov) was also evaluated on porcine spermatozoa and Galleria mellonella larvae. Cz, Ov and mGR-OLI showed the strongest antimicrobial activity; their volatile components were also able to significantly inhibit the growth of tested strains. In vitro, Ov toxicity was slightly lower than Cz, while it showed no toxicity on G. mellonella larvae. In conclusion, the study confirms the importance of evaluating natural products to consolidate the idea of safe EO applications in reducing and preventing intensive livestock infections

Origanum vulgare essential oil vs. A commercial mixture of essential oils: In vitro effectiveness on salmonella spp. from poultry and swine intensive livestock

open11noSalmonella spp. represent a public health concern for humans and animals due to the increase of antibiotic resistances. In this scenario, the use of essential oils (EOs) could be a valid tool against Salmonella contamination of meat. This work compares the in vitro effectiveness of an Italian mixture of feed additives based on EOs (GR-OLI) with EO of Origanum vulgare L., recently admitted by European Food Safety Authority (EFSA) for animal use. Twenty-nine Salmonella serotypes isolated from poultry and pig farms were used to assess GR-OLI and O. vulgare EO antimicrobial propeties. O. vulgare EO was active on the disaggregation of mature biofilm, while GR-OLI was capable of inhibiting biofilm formation and disaggregating preformed biofilm. Furthermore, GR-OLI inhibited bacterial adhesion to Caco-2 cells in a dose-dependent manner. Both products showed inhibition of bacterial growth at all time points tested. Finally, the synergistic action of GR-OLI with commonly used antibiotics against resistant strains was investigated. In conclusion, the mixture could be used both to reduce the meat contamination of Salmonella spp. before slaughter, and in synergy with low doses of ciprofloxacin against resistant strains. Although EOs as feed additives are already used in animal husbandry, no scientific study has ever highlighted their real antimicrobial potential.openDi Vito M.; Cacaci M.; Barbanti L.; Martini C.; Sanguinetti M.; Benvenuti S.; Tosi G.; Fiorentini L.; Scozzoli M.; Bugli F.; Mattarelli P.Di Vito M.; Cacaci M.; Barbanti L.; Martini C.; Sanguinetti M.; Benvenuti S.; Tosi G.; Fiorentini L.; Scozzoli M.; Bugli F.; Mattarelli P

Thymbra capitata (L.) cav. and rosmarinus officinalis (L.) Essential oils: in vitro effects and toxicity on swine spermatozoa

In the last years, Essential Oils are commanding the attention of the scientific world in a variety of fields because of their peculiar characteristics. Among the reasons behind this growing interest, there is their capability of interfering with bacteria, viruses and fungi. Moreover, they can also act as effective antioxidant and spermicidal agents. The aim of this preliminary study was to analyze the effects of the Essential Oils extracted by Cuminumcyminum and Coriandrumsativum on the membranes of spermatozoa using porcine ejaculates as model, with the future prospective of possible applications in reproductive medicine. Four different concentrations of the oils mix (1:1) were tested on samples of swine spermatozoa alongside with two controls (one with and one without Penicillin). The prepared samples were incubated at 16°C (± 1°C) in a refrigerated bath, and evaluated for Viability and Acrosome Status at three different time points (24, 72 and 120 h). When compared to the control samples, the two lower tested concentrations (0.1 and 0.2 mg/ml) do not seem to alter viability nor acrosome reaction percentage. On the other hand, the two remaining concentrations impair both parameters in a concentration-dependent manner. Overall, these preliminary results prove how this Essential Oils mix can interact with the spermatozoa membranes, both cytoplasmic and acrosoma

Optimization and Validation of a High-Performance Liquid Chromatography Method for the Analysis of Hesperidin and Carvacrol for Veterinary Use

Plant-derived compounds have been recognized by the feed industry as important supplements for livestock welfare and health. In this context, Citrus aurantium L. extract and Origanum vulgare L. essential oil have been demonstrated to have strong anti-inflammatory and antioxidant effects on animals. Being the composition of plant-derived extracts extremely influenced by the environmental and growing conditions of the plants, quality control is necessary in terms of the concentration of the active compounds to assure the reproducibility of natural feed additives. The present work aimed at the validation of the extraction procedure from feed additives of Hesperidin (HES) and Carvacrol (CAR), the main active compounds of Citrus aurantium and Origanum vulgare extracts. Then, the quantification method of both the analytes was developed and validated by reversed high-performance liquid chromatography coupled with a UV detector. The validated method was tested on premixtures and final feed additives supplied by a local feed factory to supervise the production chain. The extraction method with methanol resulted to be efficient and highly reproducible, with recovery higher than 90% for both the analytes. The chromatographic method has been demonstrated to be accurate, precise (relative standard deviation percent lower than 2.06%), and linear in the tested range concentrations, with regression coefficients equal to 0.995 and 0.999 for HES and CAR respectively. The method demonstrated that the feed additives prepared by the factory by diluting the premixtures were less concentrated than what was declared on the label

Effect of Camellia sinensis L. whole plant extract on piglet intestinal ecosystem

The employment of Camellia sinensis L. whole plant extract as food supplement in livestock nutrition has been suggested in order to prevent usual livestock intestinal diseases. The aim of the present research was to test the effects of such plant extract on the composition of pig faecal microbiota. Preliminary in vitro fermentation trials evidenced in mixed pig faecal cultures, supplemented with the tested extract, an increase of total anaerobe (p=0.02) and aerobe (p=0.03) bacteria, and a decrease of clostridia (p=0.04) compared to control cultures. Afterwards we investigated in vivo the effects on piglet faecal microbiota of a diet added with 250 mg/kg of Camellia sinensis whole plant extract. A control diet without the plant extract, but added with antibiotic (sulfadiazine, trimethoprim, and tiamulin) was used for the comparison. Microbiological analyses of faecal samples collected after 60 days of the experimental feeding, evidenced a decrease of clostridia (p=0.001) and enterococci (p=0.04) counts in the faeces of animals fed with the experimental diet, compared to those fed with the control diet. These results show that the Camellia sinensis L. whole plant extract is able to reduce the number of some potential pathogenic bacteria in piglet gut and hence might improve animal health

Indagine sulle malattie parassitarie del bovino in Emilia Romagna nell'ambito dei progetti integrati mediterranei

I dati relativi alla presenza di parassiti dell'Appennino emiliano-romagnolo sono stati analizzati ai fine di individuare i fattori di rischio associati. Fasciolosi e coccidiosi sono apparse associate a fattori geografici, le strongilosi gastro-intestinali alla effettuazione di trattamenti e al regime di rotazione dei pascol