Toxoplasma gondiiis 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 ofT. gondiiinfection using a modified agglutination test (MAT), to detectT. gondiiDNA 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 ofToxoplasmatachyzoites might frequently occur in milk of seropositive goats testing positive to n-PCR on blood.

Nosema ceranae and Nosema apis are microsporidia which play an important role in the epidemiology of honeybee microsporidiosis worldwide. Nosemiasis reduces honeybee population size and causes significant losses in honey production. To the best of our knowledge, limited information is available about the prevalence of nosemiasis in Italy. In this research, we determined the occurrence of Nosema infection in Central Italy. Thirty-eight seemingly healthy apiaries (2 to 4 hives each) were randomly selected and screened from April to September 2014 (n = 11) or from May to September 2015 (n = 27). The apiaries were located in six areas of Central Italy, including Lucca (n = 11), Massa Carrara (n = 9), Pisa (n = 9), Leghorn (n = 7), Florence (n = 1), and Prato (n = 1) provinces. Light microscopy was carried out according to current OIE recommendations to screen the presence of microsporidiosis in adult worker honeybees. Since the morphological characteristics of N. ceranae and N. apis spores are similar and can hardly be distinguished by optical microscopy, all samples were also screened by multiplex polymerase chain reaction (M-PCR) assay based on 16S rRNA-gene-targeted species-specific primers to differentiate N. ceranae from N. apis. Furthermore, PCR-positive samples were also sequenced to confirm the species of amplified Nosema DNA. Notably, Nosema spores were detected in samples from 24 out of 38 (63.2%, 95% CI: 47.8-78.5%) apiaries. Positivity rates in single provinces were 10/11, 8/9, 3/9, 1/7, or 1/1 (n = 2). A full agreement (Cohen's Kappa = 1) was assessed between microscopy and M-PCR. Based on M-PCR and DNA sequencing results, only N. ceranae was found. Overall, our results highlighted that N. ceranae infection occurs frequently in the cohort of honeybee populations that was examined despite the lack of clinical signs. These findings suggest that colony disease outbreaks might result from environmental factors that lead to higher susceptibility of honeybees to this microsporidian.

Faecal samples were individually collected from pet (n=63) and zoo (n=83) birds representing 14 orders and 63 species. All the samples were examined by faecal flotation technique. In a subgroup of samples (n=75), molecular assays were also used to detectCryptosporidiumoocysts andGiardia duodenaliscysts. Overall, 35.6% of the birds harboured parasites (42.2% of zoo birds and 27% of pet birds), including Strongyles-Capillarids (8.9%),Ascaridia(6.8%), Strongyles (5.5%),G. duodenalisAssemblage A (5.3%), Coccidia (4.1%),Cryptosporidium(4%),Porrocaecum(2.7%),Porrocaecum-Capillarids (2%), andSyngamus-Capillarids (0.7%). The zoonoticG. duodenalisAssemblage A andCryptosporidiumwere exclusively found in Psittaciformes, with prevalences of 10.3% and 7.7% within this bird group. Zoo birds were more likely to harbor mixed infections (OR = 14.81) and symptomatic birds to be parasitized (OR = 4.72). Clinicians should be aware of the public health implications posed by zoonoticG. duodenalisAssemblages andCryptosporidiumspecies in captive birds.