Candida tropicalis is less commonly isolated from clinical specimens than Candida albicans. Unlike C. albicans, which can be occasionally found as a commensal, C. tropicalis is almost always associated with the development of fungal infections. In addition, C. tropicalis has been reported to be resistant to fluconazole (FLC). To analyze the development of FLC resistance in C. tropicalis, an FLC-susceptible strain (ATCC 750) (MIC ‫؍‬ 1.0 g/ml) was cultured in liquid medium containing increasing FLC concentrations from 8.0 to 128 g/ml. The strain developed variable degrees of FLC resistance which paralleled the concentrations of FLC used in the medium. The highest MICs of FLC were 16, 256, and 512 g/ml for strains grown in medium with 8.0, 32, and 128 g of FLC per ml, respectively. Development of resistance was rapid and could be observed already after a single subculture in azole-containing medium. The resistant strains were cross-resistant to itraconazole (MIC > 1.0 g/ml) and terbinafine (MIC > 512 g/ml) but not to amphotericin B. Isolates grown in FLC at concentrations of 8.0 and 32 g/ml reverted to low MICs (1.0 g/ml) after 12 and 11 passages in FLC-free medium, respectively. The MIC for one isolate grown in FLC (128 g/ml) (128 R) reverted to 16 g/ml but remained stable over 60 passages in FLC-free medium. Azole-resistant isolates revealed upregulation of two different multidrug efflux transporter genes: the major facilitators gene MDR1 and the ATP-binding cassette transporter CDR1. The development of FLC resistance in vitro correlated well with the results obtained in an experimental model of disseminated candidiasis. While FLC given at 10 mg/kg of body weight/ day was effective in reducing the fungal burden of mice infected with the parent strain, the same dosing regimen was ineffective in mice infected with strain 128 R. Finally, the acquisition of in vitro FLC resistance in strain 128 R was related to a loss of virulence. The results of our study elucidate important characteristics and potential mechanisms of FLC resistance in C. tropicalis.

ABSTRACT. No direct evidence that genetically modified (GM) food may represent a possible danger for health has been reported so far; however, the scientific literature in this field is still quite poor. Therefore, we carried out an ultrastructural morphometrical and immunocytochemical study on hepatocytes from mice fed on GM soybean, in order to investigate eventual modifications of nuclear components of these cells involved in multiple metabolic pathways related to food processing. Our observations demonstrate significant modifications of some nuclear features in GM-fed mice. In particular, GM fed-mice show irregularly shaped nuclei, which generally represents an index of high metabolic rate, and a higher number of nuclear pores, suggestive of intense molecular trafficking. Moreover, the roundish nucleoli of control animals change in more irregular nucleoli with numerous small fibrillar centres and abundant dense fibrillar component in GM-fed mice, modifications typical of increased metabolic rate. Accordingly, nucleoplasmic (snRNPs and SC-35) and nucleolar (fibrillarin) splicing factors are more abundant in hepatocyte nuclei of GM-fed than in control mice. In conclusion, our data suggest that GM soybean intake can influence hepatocyte nuclear features in young and adult mice; however, the mechanisms responsible for such alterations remain unknown.Key words: cell nucleus/liver/genetically modified soybean Humans have been altering the genome of animals and plants for centuries and selective breeding has been used to produce some desirable characteristics such as yield increase, quality modifications or resistance to diseases. Recently, genetic modification has become the domain of molecular biology and genetic engineering, and genetically modified (GM) organisms have been produced in which new genes have been inserted into the original genome. In particular, genetic engineering has been widely applied in agriculture, thus creating GM crops which are nowadays distributed all over the world.No direct evidence that GM food may represent a possible danger for health has been reported so far; however, the scientific literature in this field is still quite poor (Schubbert et al., 1994(Schubbert et al., , 1997(Schubbert et al., , 1998Ewen and Pustzai, 1999;Chiter et al., 2000;Edwards et al., 2000;Halford and Shewry, 2000), especially as to the possible effect of a diet involving a significant amount of GM plants.The liver is a primary site for biotransformation of the products of digestion and is strategically located between the intestinal tract and the general circulation. Moreover, it degrades and detoxifies toxic compounds received from the intestines or from the general circulation and excretes them in the bile. Finally, it synthesizes many protein components of blood plasma and exercises an important degree of control over the general metabolism. Therefore, hepatocytes

Dogs are a common feeding hosts for Ixodes ricinus and may act as reservoir hosts for zoonotic tick-borne pathogens (TBPs) and as carriers of infected ticks into human settings. The aim of this work was to evaluate the presence of several selected TBPs of significant public health concern by molecular methods in I. ricinus recovered from dogs living in urban and suburban settings in central Italy. A total of 212 I. ricinus specimens were collected from the coat of domestic dogs. DNA was extracted from each specimen individually and tested for Rickettsia spp., Borrelia burgdorferi sensu lato, Babesia spp., and Anaplasma phagocytophilum, using real-time and conventional PCR protocols, followed by sequencing. Sixty-one ticks (28.8%) tested positive for TBPs; 57 samples were infected by one pathogen, while four showed coinfections. Rickettsia spp. was detected in 39 specimens (18.4%), of which 32 were identified as Rickettsia monacensis and seven as Rickettsia helvetica. Twenty-two samples (10.4%) tested positive for A. phagocytophilum; Borrelia lusitaniae and Borrelia afzelii were detected in two specimens and one specimen, respectively. One tick (0.5%) was found to be positive for Babesia venatorum (EU1). Our findings reveal the significant exposure of dogs to TBPs of public health concern and provide data on the role of dogs in the circulation of I. ricinus-borne pathogens in central Italy.

Dogs may be useful sentinels for public health monitoring of spotted fever group rickettsioses (SFGR). The aim of this study was to determine the exposure to SFGR among dogs and feeding ticks in central Italy. A total of 344 dogs and 607 adult ticks (395 Rhipicephalus sanguineus and 212 Ixodes ricinus specimens) collected from the coats of sampled animals were included in the study. Canine serum samples were analyzed by indirect fluorescent antibody technique (IFAT) for IgG antibodies against Rickettsia conorii and Rickettsia rickettsii. All the ticks and buffy coats were processed by a PCR targeting a fragment of gltA followed by sequencing. Overall, 56 dogs (16.3%) tested positive for one or both rickettsial antigens by IFAT with endpoint titers ranging from 1:64 to 1:2048; 38 (11%) serum samples reacted against R. conorii, 46 (13.4%) reacted against R. rickettsii, and 28 (8.1%) reacted simultaneously against both rickettsial agents. All buffy coats were PCR negative. Rickettsial DNA was revealed in 39 (18.4%) I. ricinus and in 10 (2.5%) R. sanguineus specimens. The amplicons sequencing showed three SFGR, that is, R. conorii detected in 10 R. sanguineus specimens and Rickettsia helvetica and Rickettsia monacensis detected in 7 and 32 I. ricinus ticks. Nine out of the 10 R. conorii isolates were obtained from ticks collected from seronegative dogs, and one specimen from a dog tested positive for both R. rickettsii and R. conorii by immunofluorescence assay. Among the seven ticks tested positive for R. helvetica, six were recovered from the coats of seronegative dogs and one from a dog having antibodies against R. conorii; the 32 isolates of R. monacensis were obtained from 28 seronegative and 4 R. conorii/R. rickettsii-positive dogs. The results highlight the non-negligible exposure of the canine population to SFGR in the sampled areas.

The lesions observed in 16 wild boars, hunted in central Italy, led to the suspect that could be related to the infection by porcine circovirus type 2 (PCV2). The animals had macroscopic and histological lesions in the lungs, tonsils, and bronchial lymph nodes. PCV2 was detected in tissue samples by polymerase chain reaction (PCR) and immunohistochemistry and it was isolated in newborn swine kidney cell cultures. From the infected cell culture supernatant, the presence of PCV2 DNA was confirmed by real-time PCR whereas virus particles were observed by electron microscopy. These diagnostic data indicate that PCV2 can infect and cause disease in Sus scrofa subspecies other than domestic swine and it is present in the wild boar population in central Italy.