Morphological and molecular characterization of Encephalitozoon hellem in hummingbirds

Microsporidiosis was identified as a cause of enteritis in wild, migratory hummingbirds (Calypte anna). Electron microscopic examinations of parasites showed microsporidian spores with a double spore coat and a polar filament containing four to six coils, compatible with the genus Encephalitozoon. Molecular analysis of ribosomal RNA genes further identified the parasites from droppings and small intestinal segments as Encephalitozoon hellem, genotype I. Microsporidial spores were identified in 19% of droppings from C. anna, Archilochus alexandri and Selasporus sasin using Gram or modified trichrome staining methods. Since E. hellem is an opportunistic pathogen in immunocompromised humans, the pathogenic potential in avian hosts, the zoonotic potential of this parasite, and the role of birds as reservoirs needs to be further explored.     

New insights onto cardiopulmonary nematodes of dogs in Sardinia,Italy

Dog heartworms Angiostrongylus vasorum and Dirofilaria immitis cause severe parasitological diseases; the importance of these parasitosis is growing due to their health impact on animals, the possible zoonotic implications and the recent spreading across several European countries and previously non-endemic areas. The aim of this study is to update the epidemiological scenario of cardiopulmonary nematodes A. vasorum and D. immitis in dogs of Sardinia island and to perform a morphological identification of larvae by the use of the Baermann and Knott techniques respectively and the molecular characterization of the mitochondrial cytochrome c oxidase subunit I (cox1) and the second ribosomal transcribed spacer region (ITS-2) of larvae L1 of A. vasorum. In the present study, 3.4 % (5/146) of dogs resulted positive at Baermann technique for A. vasorum while 8.9 % (61/684) to D. immitis. If on one side A. vasorum can be considered an emerging parasite in Sardinia, the parasitic pressure and the risk of infection for D. immitis in the island seems to be increased compared with the recent past.     

The effects of trauma among kidnap victims in Sardinia, Italy

BACKGROUND: No study to date has investigated the effects of the trauma of being kidnapped for ransom. In the present study, we aimed to assess the general health status and the presence of post-traumatic stress disorder (PTSD) and major depression (MDD) in a sample of kidnap victims. We also focused attention on dissociative experiences and on the development of the Stockholm syndrome during captivity. METHODS: We investigated the traumatic experiences and reported general health status of 24 kidnap victims using a semistructured interview. The Structured Clinical Interview for DSM-IV was used to assess the presence of PTSD and MDD. The Dissociative Experiences Scale was also administered. RESULTS: The lifetime frequency of PTSD and MDD were 45.9% and 37.5% respectively. The Stockholm syndrome had been present in 50% of the sample during captivity. The presence of PTSD can be predicted by the number of violent experiences, whereas the number of humiliating or deprivation experiences predicts the development of the Stockholm syndrome. Subjects with both PTSD and the Stockholm syndrome reported a greater number of physical complaints at the interview. CONCLUSIONS: There is no significant connection between PTSD and the Stockholm syndrome. Both are indices of the severity of the trauma of being kidnapped, but they are associated with different aspects of the traumatic experience. The presence of both syndromes appears to have a detrimental effect on physical health.     

Age at menarche in Sardinia (Italy)

The median age at menarche in 1080 Sardinian girls (ages 11.00-16.49 years) was calculated with the status quo method and probit analysis as 12.78 +/- 0.06 years.     

Detection and molecular characterization of human rotaviruses isolated in Italy and Albania

Rotaviruses are one of the most important causes of gastroenteritis in children under 5 years old. Analysis of G and P rotavirus genotypes in circulation is crucial in evaluating the appropriacy of mass vaccination of children worldwide. Overall, 592 stool samples were collected in Tirana (Albania), the Salento peninsula (South Italy), and three different hospitals in Rome (Central Italy). Of the total samples, 31.3% were rotavirus positive in Albania, 78.3% in the Salento, and 40.3% in Rome. The samples collected in Tirana and Rome were G–P typed, whereas the samples collected in the Salento were only G typed. Overall, in Italy the most frequent combinations were G4 P[8] (54.5%), G1 P[8] (27.3%), and G2 P[4] (18.2%); in Albania they were G9 P[8] (72.1%), G4 P[8] (8.8%), G1 P[8] (5.9%), and G2 P[4] (2.9%). The prevalence in Albania of atypical combinations was 7.4% for G4 P[4] and 2.9% for G9 P[4]. Phylogenetic analysis was also performed to assess the genetic relatedness of the strains. J. Med. Virol. 82:510–518, 2010. © 2010 Wiley‐Liss, Inc.     

Dystrophic epidermolysis bullosa pruriginosa in Italy: clinical and molecular characterization

Dystrophic epidermolysis bullosa (DEB) pruriginosa (DEB-Pr) is a rare variant of DEB due to COL7A1 dominant and recessive mutations, which is characterized by severe itching and lichenoid or nodular prurigo-like lesions, mainly involving the extremities. Less than 30 patients have been described showing variable disease expression, and frequently, delayed age of onset. We report the clinical and molecular characterization of seven Italian DEB patients, three affected with recessive DEB-Pr and four with dominant DEB-Pr. In all the patients, the signs were typical of a mild DEB phenotype, until the onset of pruritus, which was followed by worsening of the clinical picture, with appearance of the distinctive lichenified lesions of DEB-Pr. Nine mutations were found in the COL7A1 gene, three of which were novel and one was de novo. DEB-Pr patients with either dominant or recessive mutations were shown to synthesize a normal or variably reduced amount of type VII collagen, which was correctly deposited at the dermal-epidermal junction. Since six of these mutations have been reported in DEB patients in the absence of intense pruritus, these data implicate a role of yet unidentified phenotype-modifying factors in the pathogenesis of DEB-Pr.     

Prevalence and molecular characterization of Giardia duodenalis from sheep in central Italy

Giardiosis in domestic ruminants is an important parasitic disease and it has been shown to impair growth in lambs, thus constituting a disease of economic concern. In Europe, surveys on the prevalence of giardiosis in sheep are limited. In order to obtain additional information on the presence of giardiosis and on the potential zoonotic role of Giardia duodenalis affecting sheep in central Italy, faecal samples of 325 native sheep from 20 farms in Abruzzo region (Italy) were examined for the presence of Giardia and the isolates were genotyped and sequenced. G. duodenalis cysts were detected in five of the 325 sheep (1.5%) (mean of 450 cysts/g) coming from two farms. The 770-bp fragment of the glutamate dehydrogenase gene and the 753-bp fragment of the -giardin gene showed 100% homology with the Assemblage AI. This work suggests for the first time in Italy that sheep carry a Giardia genotype, which can be a potential public health hazard.     

Detection and molecular characterization of bovine leukemia viruses from Jordan

Bovine leukemia virus (BLV) is distributed worldwide. BLV has many effects on the health status and productivity of infected animals and is a potential risk for humans. In this study, we aimed to investigate the presence of and genotype bovine leukemia viruses on Jordanian dairy farms. Nested PCR coupled with RFLP and direct sequencing of a partial fragment of the env gene were carried out. Two BLV genotypes were found, genotypes 1 and 6. These genotypes were identified by nested PCR-RFLP of 444 bp of the env gene by restriction digestion with HaeIII, Bcl I and Pvu II. However, BLV-Jordan-10 seems to represent an entirely new genotype in our phylogenetic analysis. The nucleotide sequence identity between these two Jordanian BLV genotypes (1 and 6) was 96.2?%. The nucleotide sequence identity between Jordanian BLV genotype 1 and other reference BLV genotype 1 strains ranged from 99?% to 99.5?%. The nucleotide sequence similarity of the Jordanian BLV genotype 6 to other BLV genotypes ranged from 90?% to 96.7?%. A neutralizing motif and CD8⁺ T-cell epitope were found in the env protein of both Jordanian isolates. In this study, we documented the presence of two BLV genotypes (1 and 6) on Jordanian dairy farms.     

Morphological,toxicological and molecular characterization of a benthic Nodularia isolated from Atlantic estuarine environments

A polyphasic study of a benthic Nodularia isolate (LEGE06071) from an Atlantic environment, specifically salt pans, was performed. LEGE06071 resembled both type strains of Nodularia sphaerocarpa and Nodularia harveyana, while ACOI00729 (purchased isolate) was identified as N. sphaerocarpa. The length and width of vegetative cells varied from 3.10 to 3.15 μm and from 3.71 to 4.25 μm, respectively, while heterocyts were 3.91–4.89 μm long and 4.20–4.74 μm wide. None of the isolates had aerotopes. The sequencing of the 16S rRNA gene from the two Nodularia isolates indicated that they belonged neither to Nodularia spumigena nor N. harveyana. Nodularin and other cyanotoxin synthesis-associated genes could not be detected, nor could nodularin production be detected by ELISA. However, MALDI-TOF analysis of LEGE06071 revealed the presence of other compounds, namely, glycolipids. Hence, toxicological screenings against Artemia nauplii, Escherichia coli and Salmonella typhimurium were performed. Toxic effects could only be observed against Artemia, with 48 h-LC₅₀ values for the aqueous and crude extract of methanol of 53.21 mg ml^?1 and 17.81 mg ml^?1, respectively. This study presents the first evidence of a non-nodularin-producing Nodularia isolate in Atlantic salt pan ecosystems and its potential ecotoxicity against Artemia sp.     

Molecular characterization of Echinococcus granulosus strains in Sardinia

Investigations were undertaken to determine the genotypes of the parasite Echinococcus granulosus that were present in livestock animals on the island of Sardinia. Liver, lung, and spleen samples were obtained from 770 sheep, 229 cattle, and 277 pigs slaughtered in Sardinia between January 2003 and April 2005, and the number and fertility of hydatid cysts were determined. Protoscoleces and/or germinal layer were collected from individual cysts, DNA was extracted from 91 samples, and polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) methods were used for identification of the strain genotype for each sample (G1, G5, G6/G7). Fragments of the mitochondrial cytochrome c oxidase subunit 1 and NADH dehydrogenase I were sequenced. Hydatid disease prevalence of 75.3, 41.5, and 9.4% were found in the organs collected from sheep, cattle, and pigs, respectively. Molecular analysis showed that 89 of 91 ovine, bovine, and swine cysts belonged to the G1 genotype (common sheep strain) of E. granulosus. Parasite isolates from two pigs were identified to belong to the G7 genotype (pig strain). Our results confirm the high prevalence of E. granulosus infection in livestock animals in Sardinia and reveal the presence of at least two parasite genotypes in Sardinia.     

Cercopithifilaria spp. in dogs in Sardinia Island (Italy)

A survey on Cercopithifilaria spp. was carried out on owned and kennelled dogs in Sardinia, Italy. A total of 180 dogs were sampled and tested by microscopic detection or PCR of dermal microfilariae in skin snip sediments. The overall prevalence for Cercopithifilaria spp. at both microscopy and molecular tests was 9.4 % (17/180), while 8.3 % (15/180) of dogs scored positive at microscopic detection of sediments only. Of the 225 microfilariae measured, 212 were identified as Cercopithifilaria bainae and the remaining as Cercopithifilaria sp. II. All samples were molecularly processed for specific amplification of cytochrome oxidase subunit 1 (cox1) and ribosomal 12S gene fragments. The Basic Local Alignment Search Tool analysis of the cox1 and 12S sequences here obtained showed a high nucleotide similarity (99 and 100 %, respectively) with those of C. bainae available in GenBank. In particular, cox1 haplotype I (HI; n=?14), haplotype HXVIII (n=?2), and a new haplotype, named HXIX (n=?1), differing for a single polymorphism from HI, were detected. This study reports data on the occurrence, distribution, and genetic makeup of C. bainae and Cercopithifilaria sp. II infesting dogs in Sardinia, suggesting that these filarioids are spread in areas where Rhipicephalus sanguineus sensu lato ticks occur.     

Charcot-Marie-Tooth disease: molecular characterization of patients from Central and Southern Italy

The syndrome of peroneal muscular atrophy, or Charcot-Marie-Tooth (CMT), disease represents the most common inherited peripheral neuropathy, with a prevalence of about 1 per 2500. The disease is usually transmitted in an autosomal dominant fashion, although it can display all the mendelian patterns of inheritance. The chromosome 17-linked form (CMT1a) appears to be the most common form of the disease in all the ethnic groups studied so far, Italians included, and is due to a tandem duplication in 17p11.2. In order to study the distribution of CMT types and to establish a genotype-phenotype correlation in patients from Central and Southern Italy, we collected 19 CMT pedigrees diagnosed in the years 1992–1993. Simple tandem repeats (STR) polymorphism analysis with the marker RM11-GT and Southern blotting with the probes pVAW409R3 and pVAW412 were performed, demonstrating a high prevalence (about 60%) or 17p duplication in the families studied. No clinical or electrophysiological differences were noted between CMT1 patients with or without 17p duplication, respectively. Two families affected by CMT2 showed no evidence of rearrangement at the D17S122 locus. These data are consistent with the hypothesis of a different molecular basis for CMT2.     

A Case of Brachymetatarsia From Medieval Sardinia (Italy)

Archaeological excavations carried out in the Medieval village of Geridu (Sardinia) uncovered several burials dating to the late 13th or the first half of 14th century. Among these individuals, the skeleton of an adult female showing a bilateral abnormal shortness of the fourth metatarsal bone was identified. Bilaterality and absence of other skeletal anomalies allow to rule out an acquired aetiology of the disease and to support a diagnosis of congenital brachymetatarsia. Such a rare deformity has a clinical incidence of 0.02% to 0.05%, with strong predominance of the female gender. To our knowledge, no other cases of brachymetatarsia have been reported in paleopathology so far. Anat Rec, 297:650–652, 2014. © 2014 Wiley Periodicals, Inc.