RESUMO
The aim of this study was to assess the single and synergistic effects of fenbendazole (Fenb) and metronidazole (Metro) for the treatment of Giardia duodenalis infection in different species of non-human primates (NHPs) housed in a zoological garden of southern Italy. Moreover, the study also aimed to better define the circulation of G. duodenalis zoonotic assemblages in NHP and the potential occurrence of zoonotic transmission between the staff from the zoo and NHP. Briefly, six species that belonged to four families (Lemuridae, Cercopithecidae, Atelidae, and Hylobatidae) of NHP and housed in six cages (CG) were identified as Giardia positive and divided into two groups. Group F (N = 16 animals) was treated with Fenb (50 mg/kg, every 24 h for 5 consecutive days) and Group M (N = 7 animals) was treated with Metro (25 mg/kg, two times a day for 5 consecutive days). After the first round of therapy, all the animals were retreated for 5 days by inverting the drugs in each group. On each sampling day [study days (SDs) 3-24], the samples were tested for the presence of Giardia cysts using the FLOTAC technique. Multiple fecal tests for the antigen detection of Giardia, such as rapid ELISA and direct immunofluorescence (IFA), were performed at each sampling point only on samples that resulted in positive for Giardia cysts with FLOTAC. The efficacy of Fenb ranged from 30 to 67% and for Metro ranged from 82 to 96%. The results showed the synergistic effects of Metro and Fenb (98-100%) over the combination of Fenb and Metro (52-90%) against the infection by Giardia in NHPs. The overall k agreement between FLOTAC and IFA was reached 0.858 (p = 0.0001). In contrast, all the samples had a negative antigen result when using ELISA. At molecular analysis, six samples were confirmed positive for Giardia by nested PCR. Only two positive samples were successfully sequenced that showed 100% of identity with assemblage B. All the samples from the humans included in the study resulted in negative for Giardia cysts. Overall, the study emphasizes the need for regular monitoring of Giardia infections in NHP housed in zoos by traditional diagnostic tools combined with molecular characterization of the parasite.
RESUMO
Italy is the only European country where the crested porcupine (Hystrix cristata) lives. A parasitological investigation was performed on faecal samples, aimed to evaluate Giardia and other parasites in a free-ranging crested porcupine population in Central Italy. Samples were collected from captured and road-killed individuals as well as from feeding areas and pathways. Collected faecal samples were examined by the Mini-FLOTAC technique and a rapid immunoassay for the search of Giardia and Cryptosporidium spp. faecal antigens. For the identification of Giardia species and genotypes, molecular analysis was performed on Giardia-positive samples, by using PCR protocols able to amplify glutamate dehydrogenase, triosephosphate isomerase and a fragment of the small subunit ribosomal RNA genes. A total of 52 crested porcupine faecal samples were collected and analysed. At microscopical examination, 39 out of 52 samples were found positive for at least a single parasite species and six different parasite taxa were identified. Forty-eight percent (25/52) of faecal samples were positive for Giardia spp. and 1.9% (1/52) for Cryptosporidium spp. at the immunoassay. Among 12 faecal samples belonging to different individuals, 33.3% (4/12) were positive for Giardia spp. By using the Mini-FLOTAC technique, positivity for Trichuris spp. (32.7%, 17/52), gastrointestinal strongyles (32.7%, 17/52), capillariid eggs (3.8%, 2/52) and coccidian oocysts (1.9%; 1/52) was also evidenced. Molecular analysis was performed on 17 out of 25 Giardia-positive isolates. At the SSU rDNA locus, expected bands were achieved for 12 out of 17 isolates and all samples were assigned to Giardia duodenalis assemblage B. Sequencing at tpi locus revealed potentially zoonotic G. duodenalis assemblage AII (two isolates) and assemblage BIV (one isolate). The present study provides the first report of G. duodenalis infection in H. cristata. More in depth studies are needed on the impact and epidemiology of G. duodenalis and other identified parasites in crested porcupines.