Use of essential oils in veterinary medicine to combat bacterial and fungal infections

Essential oils (EOs) are secondary metabolites of plants employed in folk medicine for a long time thanks to their multiple properties. In the last years, their use has been introduced in veterinary medicine, too. The study of the antibacterial properties of EOs is of increasing interest, because therapies with alternative drugs are welcome to combat infections caused by antibiotic-resistant strains. Other issues could be resolved by EOs employment, such as the presence of antibiotic residues in food of animal origin and in environment. Although the in vitro antimicrobial activity of EOs has been frequently demonstrated in studies carried out on bacterial and fungal strains of different origins, there is a lack of information about their effectiveness in treating infections in animals. The scientific literature reports some studies about in vitro EOs’ activity against animal clinical bacterial and fungal isolates, but in vivo studies are very scanty. The use of EOs in therapy of companion and farm animals should follow careful studies on the toxicity of these natural products in relation to animal species and route of administration. Moreover, considering the different behavior of EOs in relation to both species and strain pathogen, before starting a therapy, an aromatogram should be executed to choose the oil with the best antimicrobial activity

Virulence and antimicrobial resistance in canine staphylococcus spp. Isolates

Dogs are reservoirs of different Staphylococcus species, but at the same time, they could develop several clinical forms caused by these bacteria. The aim of the present investigation was to characterize 50 clinical Staphylococcus isolates cultured from sick dogs. Bacterial species determina-tion, hemolysins, protease, lipase, gelatinase, slime, and biofilm production, presence of virulence genes (lukS/F-PV, eta, etb, tsst, icaA, and icaD), methicillin resistance, and antimicrobial resistance were investigated. Most isolates (52%) were Staphylococcus pseudointermedius, but 20% and 8% be-longed to Staphylococcus xylosus and Staphylococcus chromogenes, respectively. Gelatinase, biofilm, and slime production were very common characters among the investigated strains with 80%, 86%, and 76% positive isolates, respectively. Virulence genes were detected in a very small number of the tested strains. A percentage of 14% of isolates were mecA-positive and phenotypically-resistant to methicillin. Multi-drug resistance was detected in 76% of tested staphylococci; in particular, high levels of resistance were detected for ampicillin, amoxicillin, clindamycin, and erythromycin. In conclusion, although staphylococci are considered to be opportunistic bacteria, the obtained data showed that dogs may be infected by Staphylococcus strains with important virulence characteristics and a high antimicrobial resistance

Low Level of Colistin Resistance and mcr Genes Presence in Salmonella spp.: Evaluation of Isolates Collected between 2000 and 2020 from Animals and Environment

Salmonellosis is one of the most important zoonoses in Europe and the world. Human infection may evolve in severe clinical diseases, with the need for hospitalization and antimicrobial treatment. Colistin is now considered an important antimicrobial to treat infections from multidrugresistant Gram-negative bacteria, but the spreading of mobile colistin-resistance (mcr) genes has limited this option. We aimed to evaluate colistin minimum inhibitory concentration and the presence of mcr (mcr-1 to mcr-9) genes in 236 Salmonella isolates previously collected from different animals and the environment between 2000 and 2020. Overall, 17.79% of isolates were resistant to colistin; no differences were observed in relation to years of isolation (2000–2005, 2009–2014, and 2015–2020), Salmonella enterica subspecies (enterica, salamae, diarizonae, and houtenae), origin of samples (domestic animals, wildlife, and environment), or animal category (birds, mammals, and reptiles); only recently isolated strains from houseflies showed the most resistance. Few isolates (5.93%) scored positive for mcr genes, in particular for mcr-1, mcr-2, mcr-4, mcr-6, and mcr-8; furthermore, only 2.54% of isolates were mcr-positive and colistin-resistant. Detected resistance to colistin was equally distributed among all examined Salmonella isolates and not always related to the presence of mcr genes

Serological survey on the occurrence of Rickettsia spp., Neospora caninum, Bartonella henselae and Toxoplasma gondii in cats from Tuscany (central Italy)

Asymptomatic cats often harbor pathogens, some of which have not been largely investigated in feline populations. The aim of this study was to evaluate the occurrence of antibodies against Rickettsia conorii, Rickettsia felis, Rickettsia typhi, Neospora caninum, Bartonella henselae and Tox-oplasma gondii in cats from Tuscany. Ninety-five blood serum samples, previously collected, were analyzed by indirect immunofluorescence assay. Fifty-six (58.94%) cats had antibodies to at least one investigated pathogen: 28 (29.47%) cats were positive for B. henselae, 17 (17.89%) for R. felis, 14 (14.73%) for R. conorii, 14 (14.73%) for T. gondii, 2 (2.1%) for N. caninum. No cats were positive for R. typhi. Positive reactions to two or more pathogens were detected in 18 (18.94%) cats. The occurrence of antibodies against these microorganisms suggests that cats, even though asymptomatic, may be infected by pathogens, often zoonotic, and thus may be a source of infections for other animals and humans

Helichrysum araxinum Takht. ex Kirp. grown in Italy: Volatiloma composition and in vitro antimicrobial activity

In the present work the composition of biogenic volatile organic compounds (BVOCs) and the essential oil (EO) of Helichrysum araxinum Takht. ex Kirp. aerial parts, together with the antimicrobial activity, were investigated. The results showed the prevalence of sesquiterpene hydrocarbons in both spontaneous emissions as well as in the EO. The main compounds of BVOCs were γ-curcumene (10.7%), γ-muurolene (9.2%), and β-selinene (8.5%). This latter constituent also showed a similar amount in the EO and represented the most abundant compounds together with α-selinene (8.0%). It is Interesting to note the same percentage of monoterpene hydrocarbons (MHs) in both the aroma profile and the EO (18.0%) with the same most abundant compounds: β-pinene (6.3% in BVOCs vs. 5.1% in EO, respectively) and limonene (4.5% in VOCs vs. 4.9% in EO, respectively). With regard to the antimycotic activity, the EO showed to be inactive against the tested strains, while a moderate antibacterial activity was shown against Staphylococcus isolates

Survey on the presence of bacterial and parasitic zoonotic agents in the feces of wild birds

Wild avifauna may act as fecal source of bacterial and parasitic pathogens for other birds and mammals. Most of these pathogens have a relevant impact on human and livestock health which may cause severe disease and economic loss. In the present study, the fecal samples collected from 121 wild birds belonging to 15 species of the genera Anas, Tadorna, Fulica, Arddea, Larus, Falco, Athene, Accipiter, and Columba were submitted to bacteriological and molecular analyses to detect Brucella spp., Coxiella burnetii, Mycobacterium spp., Salmonella spp., Cryptosporidium spp., Giardia spp., and microsporidia. Four (3.3%) animals were positive for one pathogen: one Anas penelope for C. burnetii, one Larus michahellis for S. enterica serovar Coeln, and two Columba livia for Encephalitozoon hellem. Although the prevalence rates found in the present survey were quite low, the obtained re-sults confirm that wild birds would be the a potential fecal source of bacterial and parasitic zoonotic pathogens which sometimes can also represent a severe threat for farm animals

Antimicrobial‐Resistant Enterococcus spp. in Wild Avifauna from Central Italy

Bacteria of the genus Enterococcus are opportunistic pathogens, part of the normal intestinal microflora of animals, able to acquire and transfer antimicrobial resistance genes. The aim of this study was to evaluate the possible role of wild avifauna as a source of antimicrobial‐resistant enterococci. To assess this purpose, 103 Enterococcus spp. strains were isolated from the feces of wild birds of different species; they were tested for antimicrobial resistance against 21 molecules, vancomycin resistance, and high‐level aminoglycosides resistance (HLAR). Furthermore, genes responsible for vancomycin, tetracycline, and HLAR were searched. E. faecium was the most frequently detected species (60.20% of isolates), followed by E. faecalis (34.95% of isolates). Overall, 99.02% of the isolated enterococci were classified as multidrug‐resistant, with 19.41% extensively drug‐resistant, and 2.91% possible pan drug-resistant strains. Most of the isolates were susceptible to amoxicillin/clavulanic acid (77.67%) and ampicillin (75.73%), with only 5.83% of isolates showing an ampicillin MIC ≥ 64 mg/L. HLAR was detected in 35.92% of isolates, mainly associated with the genes ant(6)‐Ia and aac(6′)‐Ie‐aph(2′’)‐Ia. Few strains (4.85%) were resistant to vancomycin, and the genes vanA and vanB were not detected. A percentage of 54.37% of isolates showed resistance to tetracycline; tet(M) was the most frequently detected gene in these strains. Wild birds may contribute to the spreading of antimicrobial‐resistant enterococci, which can affect other animals and humans. Constant monitoring is essential to face up to the evolving antimicrobial resistance issue, and monitoring programs should include wild avifauna, to

Phenotypic and genotypic resistance to colistin in E. coli isolated from wild boar (Sus scrofa) hunted in Italy

The One Health approach is not only focused on diseases and zoonosis control but also on antimicrobial resistance. As concern this important issue, the problem of plasmid-mediated colistin resistance recently emerged. Few studies reported data about colistin resistance and mcr genes in bacteria from wildlife. In this manuscript, 168 Escherichia coli isolated from hunted wild boar were tested; colistin resistance was evaluated by MIC microdilution method, and the presence of mcr-1 and mcr-2 genes was evaluated by PCR. Overall, 27.9% of isolates resulted resistant to colistin, and most of them showed a MIC value > 256 μg/mL. A percentage of 44.6% of tested E. coli scored positive for one or both genes. In details, 13.6% of isolated harbored mcr-1 and mcr-2 in combination; most of them exhibiting the highest MIC values. Interestingly, 19.6% of mcr-positive E. coli resulted phenotypically susceptible to colistin. Wild boar could be considered a potential reservoir of colistin-resistant bacteria. In the light of the possible contacts with domestic animals and humans, this wild species could play an important role in the diffusion of colistin resistance. Thus, the monitoring programs on wildlife should include this aspect

Volatilome analyses and in vitro antimicrobial activity of the essential oils from five south african helichrysum species

Helichrysum genus was used in folk South African medicine to treat various human disorders. As a part of our on-going research addressing the exploitation of South African plants belonging to this genus, five species were investigated for their volatile and antimicrobial activities. The volatile organic compounds (VOCs) and the essential oils (EOs) were analysed by gas chromatography mass spectrometry (GC-MS). Microdilution was the method used for assessing both antimycotic and antibacterial activities, which was also tested by Kirby-Bauer agar disc diffusion. Total monoterpenes (TMs) dominated the VOCs of four species (H. trilineatum (70.6%), H. edwardsii (79.3%), H. cooperi (84.5%), and H. pandurifolium (57.0%)). H. cooperi and H. edwardsii EOs showed the predominance of TMs (68.2% and 84.5%, respectively), while H. pandurifolium and H. trilineatum EOs were characterized by the prevalence of TSs (86.5% and 43.6%, respectively). H. odoratissimum EO evidenced a similar amount of both TMs (49.5%) and TSs (46.4%). Microsporum canis was more sensitive to these EOs. The lowest minimum inhibitory concentration (MIC) was observed with H. pandurifolium and H. edwardsii EOs (0.25%). H. pandurifolium and H. trilineatum had a good effect on Staphylococcus aureus (MIC 5%). These findings open new perspectives for the exploitation of these natural compounds for application in cosmetics and pharmaceutics

Sensitivity of entomopathogenic fungi and bacteria to plants secondary metabolites, for an alternative control of Rhipicephalus (Boophilus) microplus in cattle

Boophilus (Rhipicephalus) microplus is a one host hard tick widespread in warm climates worldwide, responsible for great economic losses. To avoid resistance in ticks population, induced by the repeated administration of conventional acaricides and/or the presence of residues in the environment in meat and in milk, an alternative approach can be achieved using entomopathogenic microorganisms such as fungi and bacteria, or essential oils (EOs). The aim of the present study was to evaluate the in vitro sensitivity of Beauveria bassiana, Metarhizium anisopliae, Scopulariopsis sp, Bacillus thuringiensis and Proteus mirabilis to Eucalyptus globulus, Lavandula hybrida, Pelargonium graveolens EOs and to their main constituents such as lynalool, linalyl-acetate, geraniol, citronellol and 1,8 cineole. EOs has been chemically characterized by GC-MS. Fungal isolates were tested by a microdilution assay to achieve minimal inhibitory concentration (MIC) of both EOs and main components. The sensitivity of bacteria was evaluated by an agar disk diffusion. The results obtained show the feasibility of an integrate approach for an eco-friendly control of R. microplus by use of both entomopathogenic fungi and P. graveolens EO. L. hybrida could be an interesting alternative when B. bassiana is not employed. Conversely, a combined use of B. thuringiensis and EOs would not be advisable in the integrate control of ticks