Training in Honey Bee Veterinary Medicine in Italy: An Observational Study and Practical Proposals to Face Professional Challenges.

Honey bees, like other livestock, may be affected by infectious, parasitic, and abiotic diseases that need proper sanitary monitoring and control. Currently, there are limited opportunities for undergraduate students to receive education in Honey Bee Veterinary Medicine (HBVM) as part of their regular degree program, despite the professional requirements for veterinarians to carry out the increasing tasks related to honey bee health and production. Additionally, postgraduate training and specialization in HBVM is also underdeveloped. This study was an observational survey that evaluated the educational opportunities available in HBVM for current and future veterinarians in Italy. The survey analyzed both undergraduate and postgraduate programs, including Undergraduate Degree Programs in Veterinary Medicine (UDPVM), "Scuole di Specializzazione", Masters, and other postgraduate courses. The results indicate that the current training available for veterinarians in the field of apiculture, both before and after graduation, is also insufficient in Italy, as already reported in other EU- and extra-EU countries. Finally, a roadmap for veterinary training in HBVM is developed here describing objectives and teachings aimed at fulfilling the needs of the profession in the field of beekeeping, considering the existing rules and regulations governing public health and possible evolution of this legal framework in the future.

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.

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.

Activity of Salvia dolomitica and Salvia somalensis Essential Oils against Bacteria, Molds and Yeasts.

Essential oils (EOs) from Salvia dolomitica and Salvia somalensis, widely employed in the cosmetic and perfume industry, were analyzed for composition and tested against bacterial and fungal pathogens isolated from clinical and environmental specimens. The analyses were carried out against Staphylococcus aureus, Staphylococcus pseudointermedius, Pseudomonas aeruginosa, Escherichia coli, Streptococcus canis, Streptococcus pyogenes, Klebsiella pneumoniae, Proteus mirabilis, Microsporum canis, Microsporum gypseum, Trichophyton mentagrophytes, Aspergillus niger, Aspergillus flavus, Candida albicans, Candida krusei, Mucor sp. and Trichothecium roseum. Both EOs showed similar percentages of total monoterpenes and sesquiterpene hydrocarbons. The main constituents were 1,8-cineole and ß-caryophyllene in S. dolomitica and bornyl acetate and camphor in S. somalensis. The selected EOs have no relevant antifungal or antibacterial activities if compared to conventional drugs.

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

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.

Toxic metal levels in cocoa powder and chocolate by ICP-MS method after microwave-assisted digestion.

The Commission Regulation (EC) Regulation N. 488/2014, established the concentration limits for cadmium in specific products based on cocoa and chocolate products as from January 2019. Based on this information there is a need to determine ultratrace levels of elements that might be presents in cocoa and chocolate products. In this work, the concentrations of Arsenic, Antimony, Cadmium, Chromium, Lead, Selenium and Vanadium were evaluated in cocoa powder and chocolate by the validation of an ICP-MS method. Good selectivity/specificity, recovery, repeatability and within-laboratory reproducibility, LOD, LOQ, range of linearity, standard measurement uncertainty parameters for method validation were achieved, in accordance with Commission Regulation. The cocoa powder revealed the maximum metal concentrations of 0.303 ±â€¯0.035 mg/kg for cadmium, 1.228 ±â€¯0.146 mg/kg for lead and 0.094 ±â€¯0.013 mg/kg for arsenic. A significant difference was found between cocoa powder and chocolate samples (p < .05).

Encephalitozoon cuniculi in rabbits: Serological screening and histopathological findings.

Serological prevalence of E. cuniculi infection was assessed in 183 rabbits from central Italy. In seropositive deceased rabbits, histopathological lesions were also evaluated. Sera from 118 rabbits from 6 intensive farms, 10 rabbits from 6 family farms, 16 rabbits from a zoo, 30 rabbits from 5 research laboratories and 9 pet rabbits from 9 different owners, were tested by an enzyme-linked immunosorbent assay. Data were statistically analysed. Tissue samples from brain and kidney of 10 deceased rabbits were formalin-fixed and subsequently analysed by histopathology and immunohistochemistry. Anti-E. cuniculi antibodies were found in 129/183 (70.5%) analysed sera. At statistical analysis, E. cuniculi seropositivity was significantly higher (p<0.05) in industrial and zoo rabbits. At histology, different degrees of pathological lesions were found in serological positive (9) deceased animals. In three rabbits deceased after showing neurological signs, the severity of the lesions was interpreted as a likely cause for their death.

Seroprevalence of Toxoplasma gondii and Neospora caninum in red deer from Central Italy.

Neospora caninum and Toxoplasma gondii are cosmopolite protozoan parasites impacting on human and animal health. In particular, T. gondii commonly infects human beings and all warm-blooded animals, while N. caninum is responsible for bovine abortion and neuromuscular disease in dogs. The aim of the presented survey was to evaluate the occurrence and prevalence of these parasites in the most numerous Italian red deer population. The sera of 60 red deer (Cervus elaphus) inhabiting Central Italy (43°56'N 10°55'E) and killed by selective hunting were examined using an indirect fluorescent antibody test (IFAT) for both N. caninum and T. gondii antibodies. White blood cells (buffy coat) were also checked by PCR and T. gondii DNA was genotyped. Thirteen out of 60 sera (22%) scored positive for Toxoplasma, 17 samples (28%) were Neospora positive. Coinfection was recorded in 5 cases (8%). T. gondii (genotype II) and N. caninum DNA was detected in one and 3 samples of buffy coat, respectively. The presented study is the first to examine the occurrence of these parasites in the most numerous red deer Italian population, confirming this animal species as carrier of the investigated pathogens. These animals spread near human settlements, co-inhabiting with final hosts of T. gondii and N. caninum and could contribute to their transmission to domestic ruminants and humans. In particular, the seroprevalence value for N. caninum was the highest among European records.

Occurrence of moulds from bee pollen in Central Italy--A preliminary study.

The present study aimed to preliminary evaluate the occurrence of fungi in 40 specimens of trapped pollen collected from April-September 2015 in 40 apiaries from Tuscany (Central Italy). Cultural and microscopical examinations allowed the recognition of Cladosporium sp., Alternaria sp., Humicola sp. Mucoraceae and Acremonium sp. Penicillium brevicompactum, Penicillium chrysogenum, Aspergillus flavus , Aspergillus nidulans, Aspergillus niger and Aspergillus terreus were also identified. Yeasts and Fusarium spp. were not isolated. All pollen specimens were positive for at least one fungal isolate. Total CFU per gram ranged from 4-568. Aspergillus and Penicillium were obtained from 8 (20%) and from 22 (55%) pollen samples, respectively, associated in 4 cases (10%). The recovery of storage fungi such as Aspergillus and Penicillium in trapped pollen presents a potential risk for human health and attention should be paid to all stages of the post-harvest process.

Detection and genotyping of Toxoplasma gondii DNA in the blood and milk of naturally infected donkeys (Equus asinus).

BACKGROUND: Toxoplasma gondii is a worldwide zoonotic protozoan. Consumption of raw milk from infected animals is considered a risk factor for acquiring toxoplasmosis in humans. Recently, donkey milk has been indicated for therapeutic and nutritional purposes and T. gondii infection is common in donkeys. The purpose of the present paper was to detect the presence of parasite DNA in milk of T. gondii positive donkeys. FINDINGS: Antibodies to T. gondii were found in 11 out of 44 healthy lactating donkeys by IFAT. T. gondii DNA was detected by PCR in blood of 6 and milk of 3 seropositive jennies. Results of limited RFLP-PCR genotyping indicated the presence of T. gondii genotype II or III, commonly found in Europe. CONCLUSIONS: The occurrence of T. gondii DNA in milk suggests that the consumption of raw milk from seropositive donkeys could be a potential source of human infection.

Seroprevalence, detection of DNA in blood and milk, and genotyping of Toxoplasma gondii in a goat population in Italy.

Toxoplasma gondii is the causative agent of a major zoonosis with cosmopolitan distribution and is known to be transmitted mainly by the ingestion of undercooked or raw animal products. Drinking unpasteurized goat's milk is a risk factor associated with human toxoplasmosis. However, very little is known about the excretion of DNA in goat milk. Aim of the present study was to determine the seroprevalence of T. gondii infection using a modified agglutination test (MAT), to detect T. gondii DNA by nested-PCR (n-PCR) in samples of blood and milk from seropositive goats, and to genotype DNA isolates using 11 molecular markers in 127 adult lactating goats from 6 farms in Italy. Positive MAT results were found in 60.6% of goats while 13% of blood and milk samples from seropositive goats were positive to n-PCR. A kappa coefficient of 1 indicated a perfect agreement between blood and milk n-PCR. Genetic characterization of isolates revealed the occurrence of genotype III (n = 7), genotype I (n = 1), and atypical genotypes with hints for genotype I (n = 2). Our results suggest that the risk of excretion of Toxoplasma tachyzoites might frequently occur in milk of seropositive goats testing positive to n-PCR on blood.