Federica Berrilli

Giardia and Cryptosporidium and public health: the epidemiological scenario from the Italian perspective.

Giardia and Cryptosporidium spp. are protozoa that cause human and animal disease worldwide and often exhibit zoonotic transmission. This review gives ample information concerning the epidemiology of these parasites in Italy, i.e. prevalence data in humans, farm and pet animals, shellfish and aquatic environment. Moreover, it reports genotyping results obtained from different isolates, with particular emphasis on the spread of host-specific and zoonotic species/genotypes of various origin, and on molecular data that make the Italian situation different from that of other countries. Finally, possible explanations are given for the infrequent reports of Giardia and Cryptosporidium spp. outbreaks, despite widespread faecal contamination by these parasites.

Ribosomal and mitochondrial DNA sequence variation in Sarcoptes mites from different hosts and geographical regions

Abstract. In order to investigate the extent of the genetic variation in the DNA sequences of Sarcoptes scabiei, mite populations collected on Alpine chamois (Rupicapra rupicapra), Pyrenean chamois (Rupicapra pyrenaica) and red fox (Vulpes vulpes) from different localities of Italy and Spain were studied. Sequence analyses were carried out on the second internal transcribed spacer of the nuclear ribosomal DNA and on the 16S mitochondrial rRNA gene. ITS-2 sequences showed a higher degree of genetic polymorphism, mostly randomly distributed in the isolates from different hosts and localities, although both genomic regions are characterised by fixed nucleotide substitutions which were able to discriminate the mites collected on Pyrenean chamois from north-western Spain and on foxes from north-eastern Spain and from north-western Italy with respect to the other isolates. These results suggest the existence of a limitation to free gene exchange between the studied populations, probably related to the genetic structuring of local populations rather than to a differential adaptation to host species.

Composition and structure of helminth communities in eels Anguilla anguilla from Italian coastal lagoons

The composition and diversity of total and intestinal helminth component and infracommunities were determined in eels Anguilla anguilla from four shallow coastal lagoons near Rome. The lagoons differed principally in respect of their salinity. Only Lake Burano still received an input of freshwater, and both freshwater and marine helminth species were found in eels. In the other three lagoons all helminth species were marine or euryhaline and communities were dominated by digeneans. At component community level these three lagoons were far more similar to each other than to Lake Burano, but this distinction almost disappeared at infracommunity level. Species richness and diversity declined with increasing salinity at component community level but not at infracommunity level. Despite the changes and differences in helminth community composition, intestinal helminth infracommunity structure was very similar in all four lagoons and to that in eels from freshwater localities in Britain. The findings thus provide further and unexpected support for the view that some factor(s) other than supply side ones must be operating to produce the fundamental structural similarity in helminth communities in eels.

A study of the prevalence and genotypes of Giardia duodenalis infecting kennelled dogs.

Giardia duodenalis is a protozoan parasite of animals that is zoonotic. Given the capacity of this organism to spread via the faecal-oral route, animals held in overcrowded and unhygienic conditions are at high risk of infection. Faecal samples from dogs in three kennels in Rome were examined by microscopy and PCR for G. duodenalis, and the prevalence data generated were correlated with variables such as kennel identity, age of dog, length of time the dog had been kennelled and clinical signs. The overall prevalence of the parasite in the faecal samples was 20.5% and was higher in samples from the largest kennel, which had the greatest turnover of dogs, and in faecal samples from younger animals. Giardia cysts were found more frequently in diarrhoeic animals but were also found in dogs with no clinical signs. Although the finding that the majority of isolates were dog-specific rather than zoonotic genotypes suggests that the zoonotic risk from this pathogen is less than previously thought, the higher prevalence of infection in younger dogs may pose a specific public health issue as such animals are more frequently re-homed with families.

Acanthamoeba T4 and T15 genotypes associated with keratitis infections in Italy

Thus far there is little data available concerning Acanthamoeba associated amoebic keratitis (AK) from Italy. In order to understand the incidence of Acanthamoeba in patients with ocular infections and to characterize the isolates at the molecular level, ocular specimens and contact lenses or lens case solutions from 140 patients were analysed by culture and by an 18S rRNA (Rns) gene-based PCR method. Nineteen (13.6%) patients showed Acanthamoeba culture positive samples. Eleven out of the 14 genetically characterized isolates were assigned to the T4 genotype. Three isolates, two of them from patients with keratitis responding to specific anti-Acanthamoeba therapy, were identified as belonging to the T15 genotype. This finding represents the first association between the T15 genotype and human amoebic keratitis. PCR amplification of the 18S ribosomal DNA proved to be a sensitive method, potentially able to detect Acanthamoeba without the need of long culture incubation, and thus considerably useful for clinical applications.

Giardia cysts and Cryptosporidium oocysts in membrane-filtered municipal wastewater used for irrigation

ABSTRACT A wastewater tertiary treatment system based on membrane ultrafiltration and fed with secondary-treated municipal wastewater was evaluated for its Giardia cyst and Cryptosporidium oocyst removal efficiency. Giardia duodenalis (assemblages A and B) and Cryptosporidium parvum were identified in feed water but were found in filtered water only during occasional failure of the filtration system.

Investigation of Toxoplasma gondii presence in farmed shellfish by nested-PCR and real-time PCR fluorescent amplicon generation assay (FLAG)

To evaluate the presence of Toxoplasma gondii in edible farmed shellfish, 1734 shellfish specimens i.e., 109 Crassostrea gigas (6 pools), 660 Mytilus galloprovincialis (22 pools), 804 Tapes decussatus (28 pools) and 161 Tapes philippinarum (6 pools), were collected from the Varano Lagoon (Apulia, Italy). Shellfish from 62 pools were subjected to two molecular techniques: a nested-PCR assay, and a fluorescent amplicon generation (FLAG) real-time PCR assay, both based on the multi-copy B1 target, were performed. One pooled sample of gills from C. gigas and one pooled sample of haemolymphs from T. decussatus were assessed as positive for T. gondii DNA by both techniques. The results demonstrated the presence of T. gondii in edible farmed C. gigas and T. decussatus and indicate that there may be a considerable health threat involved in eating contaminated raw shellfish.

Helminth communities in eels Anguilla anguilla from Adriatic coastal lagoons in Italy.

The composition and diversity of the total and intestinal component and infra-communities were determined in eels Anguilla anguilla from three shallow lagoons on the Adriatic coast of Italy to determine whether the helminth communities would differ in composition and structure from those in eels from lagoons on the Tyrrhenian coast. The lagoons differed in respect of their management regimes and the extent of freshwater influx. Both freshwater and marine species of helminths were found in the eels in all three lagoons, but the freshwater component was richer in Valle Figheri. A suite of three digenean eel specialist species occurred in all three lagoons, of which any two members dominated each community. This conferred a high degree of similarity between the communities of the three lagoons. The same three species also dominated helminth communities in eels in lagoons along the Tyrrhenian coast of Italy, and compositional similarity levels were similar within and between western and eastern groups. Species richness was higher in the component communities of the eels of the Adriatic lagoons when compared to the Tyrrhenian ones, but diversity and dominance indices were of a similar order of magnitude and range. Intestinal helminth communities were richer and more diverse in two of the Adriatic lagoons because the proportion of eels with zero or one helminth species was, unusually, in the minority. It was nevertheless concluded that infracommunity structure was similar in eels from both western and eastern lagoons and that the hypothesis that it would differ in Adriatic lagoons could not be supported. The findings provide further evidence of the similarity in composition and structure of helminth communities in eels from coastal lagoons throughout Europe.

Giardia and Cryptosporidium in inflowing water and harvested shellfish in a lagoon in Southern Italy.

Giardia and Cryptosporidium spp. are important enteric protozoan pathogens for humans and animals, and have been found to contaminate water as well as edible shellfish all over the world. This is the first study to simultaneously investigate the presence of Giardia and Cryptosporidium in inflowing water and harvested shellfish in a geographically closed environment (Varano Lagoon, Southern Italy). Samples of treated wastewater were collected each month - at the outlet from the treatment plant, and downstream at the inlet into the lagoon - from the channels flowing into the Lagoon, together with specimens of Ruditapes decussatus and Mytilus galloprovincialis from shellfish-farms on the same lagoon. Giardia cysts were found by immunofluorescence (IF) microscopy in 16 out of 21 samples of treated wastewater and in 7 out of 21 samples from downstream water channels, and viable cysts were also detected by a beta-giardin RT-PCR. G. duodenalis Assemblages A and B were identified by small ribosomal subunit (18S-rDNA) and triosephosphate isomerase (tpi)-PCR, followed by sequencing. Cryptosporidium oocysts were found by IF in 5 out of 21 wastewater samples, and in 8 out of 21 samples from water channels. Molecular analysis identified the zoonotic species Cryptosporidium parvum by oocyst wall protein (COWP)-PCR and sequencing. Higher concentrations of Giardia cysts than Cryptosporidium oocysts were registered in almost all wastewater and water samples. IF and molecular testing of shellfish gave negative results for both protozoa. Wastewaters carrying Giardia and Cryptosporidium (oo)cysts are discharged into the Lagoon; however, the shellfish harvested in the same environment were found to be unaffected, thus suggesting that physical, ecological and climatic conditions may prevent contamination of harvested shellfish.

Composition and diversity of helminth communities in eels Anguilla anguilla in the River Tiber: long-term changes and comparison with insular Europe

Most studies of helminth communities in the European eel Anguilla anguilla have been undertaken in the British Isles, and there are very few analyses of community composition and structure from continental Europe. To fill this gap and test the hypothesis that helminth communities in freshwater eels in the British Isles are not typical of those of continental Europe, helminth communities of eels in the River Tiber below Rome were analysed by season using data collected in 1980 and new data from 1996. The intestinal helminth communities in the Tiber eels were species poor and characterized by low diversity. Most eels harboured one or no parasite species and communities were heavily dominated by the acanthocephalan Acanthocephalus clavula. Intestinal helminth infracommunity richness and diversity did not differ between seasons within a year or between the same seasons in 1980 and 1996, although some changes in composition were apparent. Intestinal infracommunities from Tiber eels were very similar in characteristics to those analysed from the British Isles, and their temporal changes also showed close similarities to those reported from rivers in the UK. It seems likely therefore that conclusions derived from British studies can be applied to helminth communities of eels on the continent.