Valentina Virginia Ebani

Prevalence of Leptospira and Brucella antibodies in wild boars (Sus scrofa) in Tuscany, Italy

Five hundred sixty-two blood samples were collected from wild boars (Sus scrofa) shot in six districts of Tuscany, central Italy, between 1997 and 2000. Sera were examined for antibodies specific for Leptospira interrogans by microagglutination test and Brucella spp. by the Rose Bengal test and indirect enzyme-linked immunosorbent assay. Thirty-four (6.0%) samples tested positive for anti-Leptospira antibodies, 29 (5.1%) sera were positive for anti-L. interrogans serovar bratislava antibodies (titres ranging from 1:100–1:400), and 5 (0.9%) sera were positive for anti-L. interrogans serovar icterohaemorrhagiae antibodies (titres 1:100). All the examined sera were negative for anti-Brucella antibodies.

Molecular Survey of Anaplasma phagocytophilum and Ehrlichia canis in Red Foxes (Vulpes vulpes) from Central Italy

During the 2007–2008 hunting season, 150 spleen samples were collected from free-ranging red foxes (Vulpes vulpes) in central Italy. The specimens were tested by two nested PCR assays to detect DNA of Anaplasma phagocytophilum, etiologic agent of granulocytic ehrlichiosis of animals and humans, and DNA of Ehrlichia canis, which causes the monocytic ehrlichiosis in canids. None of the foxes were PCR-positive for E. canis; 25 (16.6%) were positive for A. phagocytophilum. No specific gross alterations were detected at necropsy, and no histopathologic lesions found on PCR-positive spleen samples.

Antibacterial activity of essential oils, their blends and mixtures of their main constituents against some strains supporting livestock mastitis

Ten of the most known and used commercial essential oils (Cinnamomum zeylanicum L., Citrus bergamia Risso, Eucalyptus globulus Labill., Foeniculum vulgare Mill., Origanum majorana L., Origanum vulgare L., Rosmarinus officinalis L., Satureja montana L., Thymus vulgaris L. ct. carvacrol, Thymus vulgaris L. ct. thymol) were tested against six bacteria strains Staphylococcus aureus, Staphylococcus chromogenes, Staphylococcus sciuri, Staphylococcus warneri, Staphylococcus xylosus and Escherichia coli, responsible for mastitis in animals. The best results were achieved by S. montana, T. vulgaris ct. thymol and O. vulgare. Two binary mixtures of essential oils (EOs) were prepared of S. montana and T. vulgaris ct. thymol (ST) and of S. montana and O. vulgare (SO). The ST mixture exhibited the best inhibitory activity against all the tested bacterial strains. Two artificial mixtures of carvacrol/thymol (AB) and carvacrol/thymol/p-cymene (CD) were prepared and tested against all of the bacterial strains used. The results exhibited a general reduction of the inhibitory activity of mixture AB, although not reaching the inhibition of the ST and SO mixtures. However the mixture CD presented an apparent strong inhibition against S. aureus and S. sciuri. The EO mixtures and the mixture CD represent promising phytotherapic approaches against bacteria strains responsible for environmental mastitis.

Serological survey of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, and Ehrlichia canis infections in rural and urban dogs in Central Italy.

INTRODUCTION Borrelia burgdorferi sensu lato (s.l.) and Anaplasma phagocytophilum are well known zoonotic pathogens, whereas Ehrlichia canis is usually considered to be of veterinary concern, although on the basis of recent reports it also seems to be able to infect humans. OBJECTIVE The aim of the study was to determine the seroprevalence of B. burgdorferi s.l., A. phagocytophilum and E. canis in an Italian canine population, and to verify if there are differences between dogs living in urban areas and those from a rural environment. MATERIALS AND METHOD Blood sera of 1,965 dogs, 1,235 from cities and 730 from rural areas, were tested by indirect immunofluorescent assay (IFAT). RESULTS The overall seroprevalence was highest for E. canis (7.07%), followed by A. phagocytophilum (4.68%), and B. burgdorferi s.l. (1.47%). Rural dogs showed the highest seroprevalence to B. burgdorferi s.l. and A. phagocytophilum. No significant differences were observed between rural and urban E. canis-positive dogs. A low percentage (1.32%) of dogs with dual seropositivity was detected, and no triple positive reactions were observed. No significant differences were detected in the seroprevalence of the three agents in relationship to the age and gender of the dogs. Seroprevalence in the five years considered were not statistically different, except for the lowest rate for E. canis observed in 2012. CONCLUSIONS The results confirm the presence of B. burgdorferi s.l., A. phagocytophilum and E. canis in Italian dogs in both urban and rural areas. Monitoring pet dogs, which share the same environment with their owners, is useful for identifying the presence of tick-borne disease agents of both veterinary and public health significance.

Molecular survey of tick-borne pathogens in Ixodid ticks collected from hunted wild animals in Tuscany, Italy.

OBJECTIVE To determine the prevalence of zoonotic tick-borne bacteria in feeding ticks removed from hunted wild animals. METHODS PCR was executed on DNA extracted from 77 tick pools to detect Anaplasma phagocytophilum, Bartonella spp., Borrelia burgdorferi sensu lato, Coxiella burnetii and Rickettsia spp. RESULTS A total of 432 ticks were collected: 30 (6.94%) Haemaphysalis punctata, 72 (16.7%) Dermacentor marginatus and 330 (76.38%) Ixodes ricinus. For each animal one or two pools of 3 ticks of the same species was constituted. Seventy-seven tick pools were examined by PCR: 58 (75.32%) resulted infected and among them 14 (18.18%) showed co-infections. In particular, 29 (37.66%) pools were positive for Bartonella spp., 23 (29.87%) for Anaplasma phagocytophilum, 16 (20.78%) for Rickettsia spp., and 5 (6.49%) for Borrelia burgdorferi s.l. All samples were negative for Coxiella burnetii. CONCLUSIONS The results demonstrate the presence of several zoonotic tick-borne pathogens in the studied area, and underline the risk of exposure to infections for hunters not only during the outdoor activity, but also when they manipulate hunted animals infested by infected ticks.

Molecular detection of tick-borne pathogens in wild red foxes (Vulpes vulpes) from Central Italy

Spleen samples from 153 red foxes, shot during regular hunting season in the province of Pisa (Central Italy), were examined to detect DNA of Anaplasma phagocytophilum, Ehrlichia canis, Coxiella burnetii, Francisella tularensis, Hepatozoon canis and Babesia sp./Theileria sp. DNA of vector-borne pathogens was detected in 120 (78.43%; 95% CI: 71.06-84.66%) foxes. Specifically, 75 (49%; 95% CI: 40.86-57.22%) animals scored PCR-positive per H. canis, 68 (44.44%; 95% CI: 36.42-52.69%) for E. canis, 35 (22.88%; 95% CI: 16.48-30.35%) for piroplasms (Theileria annae), 3 (1.96%; 95% CI: 0.41-5.62%) for C. burnetii and 1 (0.65%; 95% CI: 0.02-3.59%) for A. phagocytophilum. No positive reaction was observed for F. tularensis. Fifty-six animals (36.6%; 95% CI: 28.97-44.76%) were positive for two or three pathogens. Red foxes result to be involved in the cycle of vector-borne pathogens that are associated to disease in dogs and humans.

Pseudomonas and Aeromonas isolates from domestic reptiles and study of their antimicrobial in vitro sensitivity

Pseudomonas and Aeromonas isolates from domestic reptiles and study of their antimicrobial in vitro sensitivity V. V. Ebani & F. Fratini & M. Ampola & E. Rizzo & D. Cerri & E. Andreani Published online: 6 August 2008 # Springer Science + Business Media B.V. 2008

Serological evidence of exposure to Ehrlichia canis and Anaplasma phagocytophilum in Central Italian healthy domestic cats.

The aim of the present survey was to estimate the seroprevalences of Ehrlichia canis and Anaplasma phagocytophilum in the Central Italian feline population. Serum samples of 560 healthy domestic cats were examined by indirect immunofluorescence assay (IFAT), considering an antibody titre of 1:40 as cut-off. Seroprevalences of 6.4% and 4.5% were found for E. canis and A. phagocytophilum, respectively. Adult, mixed breed cats showed seroprevalences higher than younger and purebred subjects, whereas no differences were observed in relation to gender and living conditions.

Isolation and identification of mycobacteria from captive reptiles

The occurrence of Mycobacterium species in clinically healthy pet reptiles was studied in Italy during the period 2004-2006. The feces samples of 223 animals were examined bacteriologically. Thirty-seven strains were isolated, in particular from 13/18 (72.2%) ophidians, 13/134 (9.7%) saurians and 11/71 (15.5%) chelonians. The isolates were classified, after HPLC analysis of bromophenacyl esters of cell wall mycolic acids, as Mycobacterium fortuitum (14 strains, 37.8%), Mycobacterium fortuitum-like (17, 45.9%), Mycobacterium peregrinum (4, 10.8%), and Mycobacterium chelonae (1, 2.7%). M. fortuitum was isolated from seven pythons, five saurians and two turtles; M. fortuitum-like from six saurians, six pythons and five turtles; M. peregrinum from four turtles; M. chelonae from one lizard. One isolate from an Iguana iguana could not be identified by HPLC analysis showing a previously unreported profile. Comparative 16S rDNA sequencing showed a low similarity with Mycobacterium triviale (97.2%) and Mycobacterium confluentis (97.1%). On the basis of such data the unidentified bacterium turned out to belong to a not yet described Mycobacterium species.

Experimental Brucella ovis infection in mouflon (Ovis musimon).

Brucella ovis was isolated for the first time in Italy in 1994 from the genital organs of two domestic rams. In subsequent years bacteriologic and serologic investigations demonstrated an increasing distribution of this disease in domestic sheep. Mouflon (Ovis musimon) occur in several hilly and mountainous areas of Italy where they can potentially contact domestic sheep. To determine if this species may have a role in the epidemiology of B. ovis, four male and four female mouflon, serologically negative for B. ovis and other Brucella spp., were infected intraconjunctivally with B. ovis strain BG1/94. Physical examinations, including collection of blood samples for serology and bacteriology, were performed weekly. The animals were euthanized 8 mo postinoculation (p.i.). Samples of retropharyngeal, parotid, and iliac lymph nodes; bone marrow; kidneys; spleen; epididymis; testicle; bulbourethral glands; seminal vesicles; uterus; and oviducts were collected from each animal as appropriate for histopathology and bacteriology. At the time of euthanasia none of the animals exhibited obvious clinical signs of brucellosis. The animals seroconverted 2 wk p.i. and became seronegative 24 wk p.i. Bacterial cultures, including hemocultures, were negative. No lesions due to B. ovis infection were revealed by histologic examinations. Brucella ovis probably did not infect mouflon and this wild sheep is not likely to play a role in the epidemiology of contagious epididymitis caused by B. ovis.