Domenico Cerri

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.

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.

Experimental infection of dogs with Borrelia burgdorferi

Four beagle dogs were inoculated subcutaneously with the BITs1 Italian strain of Borrelia burgdorferi. Only one dog became infected and B burgdorferi was isolated from its blood and urine three and four weeks after infection. B burgdorferi antibodies were detected by immunofluorescence from four to 11 weeks after infection. An uninoculated dog kept in the same run as the infected dog, developed a positive serological response, but none of the five dogs showed clinical signs.

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.

Detection of Mycobacterium avium subsp. paratuberculosis in cheeses from small ruminants in Tuscany

Paratuberculosis is an infectious disease which affects mainly domestic and wild ruminants caused by Mycobacterium avium subsp. paratuberculosis (Map). Map has been associated with human diseases like Crohn disease, type-1 diabetes, sarcoidosis, multiple sclerosis and Hashimotos thyroiditis. The aim of this study was to determine the level of Map positivity of cheeses produced in Tuscany (Italy) as an indication of human exposure to the specific pathogen. Sampling was focused on artisanal cheeses produced without commercial starter culture from raw sheep or goat milk, on small-scale farms. Samples were tested by quantitative PCR (qPCR) and culture. Map DNA was detected in 4/7 (57.14%) goat, and in 14/25 (56%) sheep cheeses by qPCR, whereas cultivation produced a positive result in only one case. This corresponded to a goat cheese that had also reacted positively by qPCR and yielded a viable Type S (sheep) strain of Map. The Map load of the tested samples based on qPCR ranged from 6×10 to 1.8×10(4)Map cells/g of cheese. The results indicate on average 56.57% and 66.6% positivity of cheese samples and farms, respectively. Hence, the type of cheeses that were analyzed within the context of this study seem to constitute a considerable source of human exposure to Map; although the question remains of whether the Map cells were present in a viable form, since positive results were almost exclusively recorded by qPCR.

A novel interpretation of the Fractional Inhibitory Concentration Index: The case Origanum vulgare L. and Leptospermum scoparium J. R. et G. Forst essential oils against Staphylococcus aureus strains

Origanum vulgare (oregano) and Leptospermum scoparium (manuka) were traditionally employed as natural remedies for infected wounds and skin injuries where Staphylococcus aureus is mainly involved. The first aim of this study was to investigate oregano and manuka essential oils (EOs) chemical compositions and evaluate their antibacterial activity (MIC, Minimum Inhibitory Concentration) against fourteen S. aureus wild strains. The second aim was to evaluate the antibacterial activities of oregano and manuka EOs mixed in different combination (FIC, Fractional Inhibitory Concentration) with an improved chequerboard technique. This allowed to avoid the usual uncertainty in the determination of MIC and FIC values and to obtain a more precise interpretation of FIC indexes (FICIs). Moreover, FICIs were discussed on the basis of a novel interpretation method to evaluate the synergistic/antagonistic effect of EOs mixtures. The most representative compounds in oregano EO were Carvacrol (65.93%), p-Cymene (9.33%) and γ-Terpinene (5.25%), while in manuka EO were Leptospermone (31.65%), cis-Calamenene (15.93%) and Flavesone (6.92%). EOs presented MIC values ranging from 1:2048 to 1:4096 v/v and FIC values ranging from 0.125 to 1. According to our interpretation, a synergistic effect (34.68%), a commutative effect (15.32%) and an indifferent effect (50.00%) and no antagonistic effect were observed. Conversely, according to two previously proposed FICI interpretation models, 1.80% synergistic effect could be observed and, respectively, 98.20% indifferent effect or 48.20% additive effect and 50.00% indifferent effect. As practical results, oregano and manuka EOs may be an effective alternative to chemotherapic drugs in staphylococcal infections and useful tools to enhance food security.

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

Occurrence of Vancomycin-Resistant Enterococci in Foods of Animal Origin Purchased in Tuscany

Enterococci are ubiquitous microorganisms, commonly found in plants, insects and wild animals, and are normal residents of the gastrointestinal tract of both animals and humans. They frequently occur in large numbers in certain foods, especially those of animal origin. Over the last 10 years, enterococci have emerged as major nosocomial pathogens, thus representing an increasingly important problem for public health, in relation to their ability to acquire resistance to some antimicrobial agents of relevance to human therapy, such as glycopeptides, particularly vancomycin. In this regard, the food chain represents a possible source of vancomycin-resistant enterococci for humans, mainly as a result of the use of avoparcin, a vancomycin-like glycopeptide, as a feed additive for growth promotion of farm animals, as undoubtedly established by previous studies (Bates, 1997; Mc Donald et al., 1997). Although the utilization of avoparcin in animal husbandry has been banned since 1997, VRE have recently been isolated from foods of different typology (Giraffa et al., 2000; Knijff et al., 2002), therefore investigation into the frequency of recovery of VRE at this level is still recommendable. Thus, the present survey was carried out with the aim of evaluating the occurrence of VRE in foods of animal origin purchased in Tuscany, considering that these microorganisms could play a relevant role both directly as potential pathogens for humans, as well as in relation to their capability to spread antibiotic resistance genes to other pathogenic bacteria, such as Staphylococcus aureus.

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.