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1.
Viruses ; 16(6)2024 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-38932224

RESUMO

Porcine parvoviruses (PPVs) are among the most important agents of reproductive failure in swine worldwide. PPVs comprise eight genetically different species ascribed to four genera: Protoparvovirus (PPV1, PPV8), Tetraparvovirus (PPV2-3), Copiparvovirus (PPV4-6), and Chaphamaparvovirus (PPV7). In 2016, PPV7 was firstly detected in the USA and afterwards in Europe, Asia, and South America. Recently, it was also identified in Italy in pig farms with reproductive failure. This study aimed to evaluate the circulation of PPV7 in domestic and wild pigs in Sardinia, Italy. In addition, its coinfection with Porcine Circovirus 2 (PCV2) and 3 (PCV3) was analysed, and PPV7 Italian strains were molecularly characterised. PPV7 was detected in domestic pigs and, for the first time, wild pigs in Italy. The PPV7 viral genome was detected in 20.59% of domestic and wild pig samples. PPV7 detection was significantly lower in domestic pigs, with higher PCV2/PCV3 co-infection rates observed in PPV7-positive than in PPV7-negative domestic pigs. Molecular characterisation of the NS1 gene showed a very high frequency of recombination that could presumably promote virus spreading.


Assuntos
Coinfecção , Infecções por Parvoviridae , Parvovirus Suíno , Filogenia , Doenças dos Suínos , Animais , Parvovirus Suíno/genética , Parvovirus Suíno/classificação , Parvovirus Suíno/isolamento & purificação , Itália/epidemiologia , Infecções por Parvoviridae/veterinária , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/virologia , Suínos , Doenças dos Suínos/virologia , Doenças dos Suínos/epidemiologia , Coinfecção/virologia , Coinfecção/veterinária , Coinfecção/epidemiologia , Genoma Viral , Circovirus/genética , Circovirus/classificação , Circovirus/isolamento & purificação , Infecções por Circoviridae/veterinária , Infecções por Circoviridae/virologia , Infecções por Circoviridae/epidemiologia , DNA Viral/genética
2.
Acta Trop ; 252: 107151, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38367944

RESUMO

Cystic Echinococcosis (CE), caused by the larval form of Echinococcus granulosus sensu lato, is a neglected zoonosis still threatening public health worldwide. In Italy different epidemiological scenarios were reported depending on the geographical area and associated socio-economic activities. Although in northern Italy the occurrence of E. granulosus is considered sporadic, in the southern regions and, particularly in Sardinia, CE prevalence reaches high levels. We analysed CE cysts collected from infected sheep from various areas of mainland Italy and the Sardinia island, with the main objective to investigate intergenotypic and intragenotypic variations at national level. CE cysts were collected from slaughtered sheep following post mortem inspection at local abattoirs. Total genomic DNA was extracted and amplification and sequencing of the partial mitochondrial genes nad5 and cox1 were performed. A Bayesian phylogenetic tree was estimated on a nad5 dataset (n = 260) composed of E. granulosus samples from this study (n = 126) and all the nad5 haplotypes available in GenBank (n = 134). In addition, haplotype network, diversity and neutrality analysis were performed on nad5 and cox1 sequences of Italian origin obtained in this study. E. granulosus sensu stricto (s.s.) was found to be the only Echinococcus species infecting sheep in Italy, mainly represented by G1 genotype (76 %) and, to a lower extent, by G3 genotype (24 %). Phylogenetic analyses revealed 40 nad5 and 33 cox 1 haplotypes, and the presence of two founder haplotypes, belonging to G1 and G3 genotype, showing 100 % similarity with DNA sequences from different geographic regions. The lack of geographical segregation, high haplotype and low nucleotide diversity coupled with significant negative values of Tajima's D and Fu's Fs observed in this study indicated high genetic variation and demographic expansion of E. granulosus s.s. in Italy.


Assuntos
Cistos , Equinococose , Echinococcus granulosus , Animais , Ovinos , Echinococcus granulosus/genética , Filogenia , Teorema de Bayes , Variação Genética , Equinococose/epidemiologia , Equinococose/veterinária , Haplótipos , Genótipo , Itália/epidemiologia
3.
Viruses ; 15(11)2023 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-38005836

RESUMO

Porcine Circovirus type 2 (PCV2) is the etiological agent of a disease syndrome named Porcine Circovirus disease (PCVD), representing an important threat for the pig industry. The increasing international trade of live animals and the development of intensive pig farming seem to have sustained the spreading of PCVD on a global scale. Recent classification criteria allowed the identification of nine different PCV2 genotypes (PCV2a-i). PCV2a was the first genotype detected with the highest frequency from the late 1990s to 2000, which was then superseded by PCV2b (first genotype shift). An ongoing genotype shift is now determining increasing prevalence rates of PCV2d, in replacement of PCV2b. In Italy, a complete genotype replacement was not evidenced yet. The present study was carried out on 369 samples originating from domestic pigs, free-ranging pigs, and wild boars collected in Sardinia between 2020 and 2022, with the aim to update the last survey performed on samples collected during 2009-2013. Fifty-seven complete ORF2 sequences were obtained, and the phylogenetic and network analyses evidenced that 56 out of 57 strains belong to the PCV2d genotype and only one strain to PCV2b, thus showing the occurrence of a genotype shift from PCV2b to PCV2d in Sardinia.


Assuntos
Infecções por Circoviridae , Circovirus , Doenças dos Suínos , Suínos , Animais , Filogenia , Circovirus/genética , Comércio , Infecções por Circoviridae/epidemiologia , Infecções por Circoviridae/veterinária , Doenças dos Suínos/epidemiologia , Internacionalidade , Sus scrofa , Genótipo , Itália/epidemiologia
4.
Diseases ; 11(3)2023 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-37489442

RESUMO

An uncommon clinical case of an adult woman who was referred to the hospital with severe symptoms attributable to cystic echinococcosis (CE) is described in this report. According to a questionnaire, the subject was exposed to a high risk of infection since she was employed on a farm about 20 years before diagnosis. She lived close to several animal species and handled vegetables in inadequate hygienic conditions. Medical and laboratory investigations confirmed the presence of massive echinococcal cystic lesions in each lung and in the liver. Given the peculiarity of the case, pharmacological and surgical treatments were the only conceivable option. The association of pharmacological treatment, surgery, and interventional radiology procedure represented a reliable and effective way to handle a complex case of human hydatidosis. A multi-disciplinary approach was mandatory, resulting in a clear and conclusive diagnosis of CE caused by the zoonotic parasite E. granulosus sensu stricto of the G1 genotype.

5.
Diseases ; 11(3)2023 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-37489443

RESUMO

This study involved 20 patients affected by cystic echinococcosis (CE) who were referred to different hospitals of Sardinia (Italy) from 2017 to 2022. By means of a multidisciplinary approach, diagnosis was confirmed for CE in 18 patients and for different aetiologies in two subjects. Moreover, serology was positive for 15 subjects. Since multiple CE cysts were found in five patients, a total of 27 lesions were collected; however, only one for each patient was investigated for genetic characterization of E. granulosus s.s. DNA isolates. Our results included 15 fertile cysts that underwent DNA extraction and amplification by three different PCRs targeting nuclear (calreticulin) and mitochondrial genes (cox1 and nad5). DNA was sequenced, and by neighbour-joining phylogenetic trees we determined 10 G1 and five G3 genotypes previously reported in Sardinia. These sequences were used to construct a network, along with those circulating in Mediterranean areas. The haplotype network calculated on cox1 evidenced seven different haplotypes of the 15 isolates, with SAR2 the most represented, carried by seven cysts, and SAR17 never described in the Mediterranean area. Meanwhile, the nad5 sequences showed the most common haplotype as nd5SAR7, as well as two new haplotypes not previously described, nd5SAR13, isolated from a Sardinian patient, and nd5SAR14, isolated from a Romanian patient.

6.
Vet Sci ; 10(5)2023 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-37235401

RESUMO

Tapeworm infections are among the most relevant parasitic diseases in humans and animals. Tapeworms from the Genus Echinococcus are particularly important as they can cause cystic or alveolar echinococcosis. A molecular screening was performed on 279 fecal samples collected from carcasses of wild carnivores from Central Italy using PCR targeting diagnostic fragments of nad1, rrnS, and nad5 genes. Samples positive for either Taenia spp. or Echinococcus granulosus were sequenced to taxonomically identify the parasitic DNA. Of the 279 samples, 134 (48.0%) gave positive results in the multiplex PCR. Only one (0.4%) sample from an Apennine wolf tested positive for Echinococcus granulosus sensu stricto (genotype G3), whereas no sample tested positive for E. multilocularis. The most frequently detected tapeworms were: Mesocestoides corti (syn M. vogae) (12.9%), M. litteratus (10.8%), Taenia serialis (9.3%), and T. hydatigena (6.5%), other tapeworms were rarely detected. The results suggest that Echinococcus infections in Central Italy do not seem to be sustained by sylvatic cycles, confirming the absence of E. multilocularis in Central Italy. The survey corroborates, yet again, the importance of passive surveillance of wild animals that can serve as reservoirs for zoonotic pathogens, especially on wild canids that in other areas are strongly implicated in the transmission of E. granulosus and E. multilocularis.

7.
Int J Parasitol Parasites Wildl ; 21: 11-16, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37025622

RESUMO

Tapeworms are trophically-transmitted and multi-host parasites with a complex indirect life cycle, strictly depending on predator-prey interactions. Their presence in a free-living population, mainly definitive hosts, is arduous to study due to the complexity of collecting fecal samples. However, epidemiological studies on their frequency are crucial from a public health perspective, providing information on food habits and prey selection of predators. The present study aims to update the frequency of tapeworms detected in stool samples by molecular analysis in Italian wolf populations of Umbria and Marche regions collected from 2014 to 2022. Tapeworm's total frequency was 43.2%. In detail, Taenia serialis was detected in 27 samples (21.6%), T. hydatigena in 22 (17.6%), and Mesocestoides corti (syn. M. vogae) in 2 (1.6%). Three samples were identified as M. litteratus and E. granulosus s.s. (G3) and T. pisiformis, with a proportion of 0.8%, respectively. The low frequency of E. granulosus in a hyperendemic area is discussed. The results show for the first time a high frequency of Taenia serialis not comparable to other Italian studies conducted on wild Carnivora; thus, a new ecological niche is conceivable. These findings suggest a plausible wolf-roe deer cycle for T. serialisin the investigated area.

8.
Viruses ; 15(2)2023 01 18.
Artigo em Inglês | MEDLINE | ID: mdl-36851491

RESUMO

Understanding how geography and human mobility shape the patterns and spread of infectious diseases such as COVID-19 is key to control future epidemics. An interesting example is provided by the second wave of the COVID-19 epidemic in Europe, which was facilitated by the intense movement of tourists around the Mediterranean coast in summer 2020. The Italian island of Sardinia is a major tourist destination and is widely believed to be the origin of the second Italian wave. In this study, we characterize the genetic variation among SARS-CoV-2 strains circulating in northern Sardinia during the first and second Italian waves using both Illumina and Oxford Nanopore Technologies Next Generation Sequencing methods. Most viruses were placed into a single clade, implying that despite substantial virus inflow, most outbreaks did not spread widely. The second epidemic wave on the island was actually driven by local transmission of a single B.1.177 subclade. Phylogeographic analyses further suggest that those viral strains circulating on the island were not a relevant source for the second epidemic wave in Italy. This result, however, does not rule out the possibility of intense mixing and transmission of the virus among tourists as a major contributor to the second Italian wave.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , COVID-19/epidemiologia , Epidemiologia Molecular , Itália/epidemiologia , Filogeografia , Variação Genética
9.
J Infect Dev Ctries ; 17(12): 1814-1820, 2023 12 31.
Artigo em Inglês | MEDLINE | ID: mdl-38252735

RESUMO

INTRODUCTION: Humans acquire cystic echinococcosis through accidental ingestion of Echinococcus granulosus (EG) eggs released into the environment by infected dogs. This study aimed to determine the presence of EG antibodies and their determinants in owned dogs in Ibadan, Nigeria. METHODOLOGY: Sera from 185 dogs on routine visits to veterinary clinics were analysed by indirect ELISA. A questionnaire was administered to dog owners to obtain data on demographics, management, and environmental factors. Data were analysed using descriptive statistics, univariate analysis, and logistic regression at α0.05. RESULTS: The median age of the dogs was 20 months (range 2 - 96). The seroprevalence of EG infection was 33.51% (95% CI: 26.71, 40.32%). Low educational level of dog owners (OR: 2.8; 95% CI: 1.3, 5.8); local dog breeds (OR: 3.2; 95% CI: 1.7, 6.0); confinement (OR: 0.4; 95% CI: 0.2, 0.8); interaction with other dogs (OR: 3.2; 95% CI: 1.4, 7.3); self-dewormed dogs (OR: 2.6; 95% CI: 1.2, 5.9) and never dewormed dogs (OR: 4.39; CI: 1.9, 10.0) were significantly associated with EG seropositivity. Our results suggest also that local breed of dog (AOR: 2.4; 95% CI: 1.2, 4.9), self-deworming of dogs (AOR: 2.6; 95% CI: 1.1, 5.9) and the absence of any dog deworming treatment (AOR: 2.9; 95% CI: 1.2, 7.1) might be predictive of EG seropositivity. CONCLUSIONS: Our study provides evidence of EG infection in owned dogs, especially in those medicated by owners. Deworming practices should be based on the recommendations of a veterinarian to effectively prevent EG transmission from dogs to humans.


Assuntos
Equinococose , Echinococcus granulosus , Humanos , Cães , Animais , Lactente , Pré-Escolar , Criança , Estudos Soroepidemiológicos , Nigéria/epidemiologia , Equinococose/epidemiologia , Equinococose/veterinária , Anticorpos Anti-Helmínticos
10.
Vet Sci ; 9(12)2022 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-36548840

RESUMO

Environmental contamination with parasite eggs poses a serious risk to public health. This study aimed to assess the presence of taeniid eggs and, in particular, E. granulosus s.l., in environmental samples in the city of Ibadan, South West Nigeria. To this purpose, soil (n = 200), fecal (n = 200) and water samples (n = 50) were examined by microscopic observation and the multiplex PCR method. The influence of specific environmental factors on E. granulosus s.l. egg dispersion was also evaluated. Taeniid eggs were microscopically found in 11.5%, 25.5% and 8.0% of soil, fecal and water samples, respectively. PCR analyses evidenced the presence of E. granulosus s.l. in 8.0%, 24.0% and 2.0% of soil, fecal and water samples, respectively. The proximity to slaughterhouses, the level of urbanisation and the local government area of belonging did not seem to affect E. granulosus s.l. egg dissemination patterns. Our results have clearly demonstrated that both urban and semi-urban areas of the city of Ibadan in Nigeria are highly contaminated by taeniid eggs and we recommend the adoption of appropriate measures to control E. granulosus s.l.

11.
Comp Immunol Microbiol Infect Dis ; 86: 101816, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35472655

RESUMO

Cystic echinococcosis (CE) is a disease caused by Echinococcus granulosus sensu lato (s.l.), an ubiquitous worldwide zoonotic agent affecting humans and animals. Diagnosis of CE in humans is usually performed by imagine techniques along with immunoassays. The aim of our study was to evaluate and compare four commercial diagnostic kits, based on the detection of IgG antibodies against E. granulosus and E. multilocularis. The study was performed on a total of 259 sera: the positive (n = 74) and the negative (n = 185) group. The following analytic and diagnostic performances of the four kits were evaluated: operator skills, specificity, sensitivity, repeatability, reproducibility, accuracy, positive and negative predictive values. Based on the parameters evaluated, all four tests demonstrated excellent quality and proved to be reliable diagnostic tools to support the clinical evaluation of human patients suspected of having CE. The four commercial assays, in our hands, presented altogether, a range of performances from good to excellent, being immunoblotting (IB) the most reliable, used as gold standard, followed by the immunochromatographic test (ICT) and finally the two enzyme linked immunosorbent assay (ELISAs).


Assuntos
Equinococose , Echinococcus granulosus , Animais , Equinococose/diagnóstico , Equinococose/veterinária , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/veterinária , Humanos , Immunoblotting/veterinária , Reprodutibilidade dos Testes
12.
Acta Trop ; 226: 106253, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34822852

RESUMO

Cystic echinococcosis (CE) diagnosis by means of serological assays is hampered by the presence of parasites closely related to Echinococcus granulosus sensu lato (s.l.), responsible of the zoonotic disease and with which share cross-reacting antigens. Thus, improvements on the characterization of Echinococcus specific antigens expressed in the larval stage are required, in order to provide useful information for the development of immunological assays for the serodiagnosis of CE in sheep. Here, the proteome of the hydatid cyst fluids (HFs) of Echinococcus granulosus (hydatid fluid, EgHF) and other ovine parasites cyst fluids (CFs), Taenia hydatigena (ThCF) and Taenia multiceps (TmCF) were analyzed by a shotgun proteomic approach. Parasite and host protein profiles in the three types of cyst fluids were characterized and compared. Among the identified proteins, differential parasitic markers with serodiagnostic potential, due to their well-known immunoreactivity in human, included Ag5, AgB proteins, 8-kDa glycoproteins, hydatid disease diagnostic antigen P29 and major egg antigen P40. In particular, seven proteoforms of AgB and 8-kDa glycoprotein resulted to be the most promising diagnostic biomarkers, as they might predict CE in ovine and discriminate between different types of parasites.


Assuntos
Echinococcus granulosus , Echinococcus , Taenia , Animais , Líquido Cístico , Proteômica , Ovinos
13.
Vet Sci ; 8(6)2021 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-34198886

RESUMO

Echinococcus granulosus sensu lato (s.l.) infection in dogs poses risk of transmission to their owners and family members. We determined the prevalence and factors associated with E. granulosus s.l. infection among owned dogs presented at veterinary clinics or hospitals in Lagos State, Nigeria. Fecal samples from 217 dogs were screened for the presence of taeniid eggs using a sedimentation test in a cross sectional study. The taeniid eggs were identified at molecular level using a multiplex PCR. A structured questionnaire was used to obtain data on intrinsic and extrinsic factors from 133 dog owners. Out of the 217 dog fecal samples, 13 (6.0%) had taeniid eggs, of which 12 (92.3%) were identified as Echinococcus granulosus s.l. We found that Echinococcus granulosus infection is present among owned dogs in Lagos State with an overall prevalence of 5.5%. Location of the veterinary clinics or hospital and purpose for keeping dogs were significant factors associated with E. granulosus infection among owned dogs. Dogs living in suburban areas and kept for security purposes or guarding have higher probability of infection. Appropriate and regular treatment of dogs with praziquantel is highly recommended to reduce risk of E. granulosus transmission to humans.

14.
Pathogens ; 10(2)2021 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-33530642

RESUMO

Echinococcus granulosus sensu lato (s.l.) is the causative agent of cystic echinococcosis in animals and humans. Different E. granulosuss.l. genotypes exhibit great diversity in their life cycle, host selectivity and pathogenicity. For this reason, the study of genetic variation within Echinococcus species is of importance for their epidemiological implication. We employed two SNP genotyping technologies to distinguish G1 and G3 E. granulosus sensu stricto (s.s.). genotypes. The genotypes of DNA samples (n = 28) extracted from hydatid cysts of different animal species were identified by amplification and sequencing of a fragment of the mitochondrial nad5 gene. Two SYBR green and three TaqMan real time PCR assays were developed for targeting of three nad5 informative positions (SNP758, 1123, and 1380) known to be able to discriminate G1 from G3. Genotyping by SYBR Green PCR based on cycle threshold (Ct) with melting temperature (Tm) analysis and performed on SNP1123 and SNP1380 failed to identify one DNA sample. TaqMan assays for SNP758, 1123 and 1380 effectively confirmed genotype identification obtained by Sanger sequencing. Our results demonstrated that the combination of the three Taqman assays developed in this study represents a valuable and cost effective tool alternative to DNA sequencing for E. granulosus s.s. genotyping.

15.
Pathogens ; 9(11)2020 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-33143032

RESUMO

Cystic echinococcosis (CE), a zoonotic disease caused by the larval stage of the tapeworm Echinococcus granulosus sensu lato (s.l.), is a worldwide public health problem. Echinococcus granulosus sensu stricto (s.s.), associated with G1 and G3 genotypes, is endemic with high prevalence in the Mediterranean basin. The parasite's life cycle comprises definitive hosts (canids) and intermediate hosts (ruminants) and can occasionally involve humans. The main aim of this research was to confirm the diagnosis of 13 patients suspected of CE who presented different complications and needed the surgical removal of the cysts. We also wanted to understand and clarify more the diagnosis of echinococcosis in humans. For this purpose, the patients first underwent cyst evaluation by ultrasound (US), immunological analysis, and then total pericystectomy, followed by parasitological, histopathological, and molecular biology examinations of the cysts. US stadiated one CE1, one CE2, eight CE3b, one CE4, and two CE5; immunology evidenced nine positives; histopathology confirmed 11 CE cysts, of which 8 fertile presenting protoscoleces were identified as E. granulosus s.s. by molecular biology, genotyped as three G1 and four G3 by neighbor-joining (NJ) phylogenetic tree. In conclusion, the results showed that 11 patients were affected by E. granulosus s.s. G1 orG3, and 2 cystic neoformations were of non-parasitic origin.

16.
Pathogens ; 9(10)2020 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-32992621

RESUMO

Diagnosis of cystic echinococcosis (CE) in sheep is essentially based on necropsy findings. Clinical symptoms can be easily overlooked, while the use of immunological tests is still not recommended for an intra vitam diagnosis. This study assessed the performances of three post-mortem laboratory methods in the diagnosis of ovine CE. In the absence of a single and accurate test as a gold standard, the results of multiple analytical tests can be combined to estimate diagnostic performance based on a Bayesian statistical approach. For this purpose, livers (n = 77), and lungs (n = 79) were sampled from adult sheep and examined using gross pathology, histopathology and molecular analyses. Data from the three diagnostic methods were analyzed using a Bayesian latent class analysis model to evaluate their diagnostic accuracy in terms of sensitivity (Se), specificity (Sp), positive predictive value (PPV) and negative predictive value (NPV). The gross pathology examination revealed excellent diagnostic capabilities in diagnosing ovine CE with an Se of 99.7 (96.7-99.8), Sp of 97.5 (90.3-99.8), PPV of 97.6 (90.5-100), and NPV of 99.7 (96.5-100). The experimental design used in this work could be implemented as a validation protocol in a quality assurance system.

17.
Pathogens ; 9(5)2020 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-32370251

RESUMO

Porcine circovirus 3 (PCV3) is a recently discovered member of the Circoviridae family. So far, its presence has been reported in North America, Asia, South America, and Europe. In this study, blood and tissue samples from 189 Sardinian suids (34 domestic pigs, 115 feral free ranging pigs, and 39 wild boars) were used to genetically characterize the PCV3 strains from Sardinia. PCV3 infection in the animals was confirmed by real time PCR. The detection rate in the three groups analyzed was l7.64% in domestic pigs, 77.39% in free ranging pigs, and 61.54% in wild boars. Moreover, our results showed that co-infection of PCV3 with other viruses is quite a common occurrence. Molecular characterization of Sardinian PCV3 strains was performed by sequencing 6 complete genomes and 12 complete cap genes. Our results revealed that there is a high similarity between our strains and those identified in different countries, confirming the genetic stability of PCV3 regardless of geographical origin. Haplotype network analysis revealed the presence of 6 whole genomes or 12 unique ORF2 haplotypes and a nonsynonymous mutation in ORF2 that leads to an R14K amino acid substitution. Phylogenetic analysis of whole genome and ORF2 was also conducted. The Sardinian strains were allocated in three different clusters of phylogenetic trees of both complete genome and ORF2. With this study, we have provided a snapshot of PCV3 circulation in Sardinia. Our findings might help to achieve a deeper understanding of this emerging porcine virus.

18.
Parasitol Int ; 77: 102120, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32259586

RESUMO

Cystic echinococcosis (CE) is a severe parasitic zoonosis caused by the metacestode of the tapeworm Echinococcus granulosus sensu lato (s.l.). The disease has a global distribution representing a significant public health concern. Based on mitochondrial DNA analysis E. granulosus s.l. has been subdivided into five species: E. granulosus sensu stricto (s.s.) (G1, G3 genotype), E. equinus (G4 genotype), E. ortleppi (G5 genotype), E. canadensis (G6-G8, G10 genotype) and E. felidis. E. granulosus s.s., and in particular G1, is the most widespread genotype and the major responsible of human CE cases worldwide. In Italy G1 genotype is higly represented with larger percentages in some hyperendemic areas such as Sardinia. Molecular studies represent a valuable tool to improve our understanding of the E. granulosus epidemiology and CE control strategies. In the present study we investigated genetic variability of E. granulosus s.s. in Sardinia. To this purpose 83 hydatid cysts were collected from different animal species including humans and the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene was partially sequenced (720 bp). Nucleotide sequences from Mediterranean basin were also analyzed for comparison. The phylogenetic network revealed 30 haplotypes grouped around a predominant isolate that had been already reported from other world regions. Haplotype diversity (0.8495 ± 0.0336) and nucleotide diversity (0.003305 ± 0.002014) were similar in Sardinia respect to other Mediterranean countries. Neutrality indices obtained by Tajima's D and Fu's Fs test were significantly negative (p ≤ .01) suggesting expansion of Sardinian population. Low Fixation indices (Fst), ranging from negative values (Algeria, Greece, Spain, other part of Italy) to 0.089 (Albania, France), indicated absence of genetic differentiation, and gene flow between Sardinia and other Mediterranean countries.


Assuntos
Equinococose/epidemiologia , Echinococcus granulosus/genética , Variação Genética , Haplótipos , Animais , Animais Domésticos/parasitologia , Ciclo-Oxigenase 1/genética , Equinococose/parasitologia , Echinococcus granulosus/classificação , Genótipo , Humanos , Itália/epidemiologia , Região do Mediterrâneo/epidemiologia , Filogenia
19.
Infect Genet Evol ; 71: 189-196, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30904672

RESUMO

Porcine circovirus type 2 (PCV2) is associated with multi-factorial syndromes, commonly known as porcine-circovirus-associated diseases, which cause severe economic losses in the swine industry worldwide. Four genotypes (PCV2a, PCV2b, PCV2c, and PCV2d) have been identified. Lately, the prevalence of PCV2d has been increasing in many countries, thereby prefiguring a global replacement of PCV2b. Wild boars are also susceptible to PCV2 infection, with virus prevalence similar to that of domestic pigs. This work was aimed at expanding the knowledge about the molecular epidemiology of PCV2 in Italy. For this purpose, we analysed 40 complete ORF-2 sequences from PCV2 strains isolated from domestic pigs and wild boars in Sardinia (Italy) over a period of 5 years (2009-2013). Phylogenetic and Bayesian analyses were performed on three data sets compiled from DNA sequences over a large geographical area. PCV2b was found to be dominant in Sardinia, whereas no PCV2a and PCV2c were found. This study indicates the presence of genotype PCV2d-2 infecting both domestic and wild pigs, thus confirming its circulation in Italy. Sardinian sequences clustered mostly with Italian isolates and with strains from China, Belgium, Croatia, Taiwan, Korea, and Portugal. Genetic variability of PCV2 in Sardinia appears to be a result of both local viral evolution and different epidemic introduction events.


Assuntos
Infecções por Circoviridae/epidemiologia , Circovirus/genética , Animais , Infecções por Circoviridae/transmissão , Genótipo , Itália , Epidemiologia Molecular , Filogenia , Sus scrofa , Suínos , Doenças dos Suínos/virologia
20.
Mol Biol Rep ; 46(2): 1747-1755, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30734170

RESUMO

The Italian National Reference Center for Echinococcosis (CeNRE, Sassari, Italy) set up a diagnostic protocol of "one-step-PCR" useful for the detection of E. granulosus sensu stricto (E.g.s.s.) and the identification of its genotype (G1-G3). The purpose of this work was to perform the validation of the "PCR E.g.s.s." method. The procedures were performed employing the criteria of the World Organization for Animal Health as well as of the Italian Accreditation Body (ACCREDIA) based on the Regulation UNI CEI EN ISO/IEC 17025. Positive DNA samples belonging to E. granulosus, E. ortleppi, E. multilocularis, E. canadensis species were used for the experiments. Analytical specificity evidenced primer pairs Cal (Calreticulin l gene of 1001 bp) with an specificity higher respect to Ef1 (Elongation-Factor 1 Alpha gene of 706 bp) and NAD (Dehydrogenase-subunit 1 gene of 219 bp). The analytical sensitivity presented the capability to detect a very low amount of parasite DNA corresponding to a concentration of 12.5 pg/µl; accuracy and precision related to the operator performance, along with repeatability and reproducibility, evidenced high concordance among results and demonstrated an excellent κ values of Cohen. According to the good performance related to the evaluated parameters, the method "PCR E.g.s.s." was suitable for the validation procedure, and consequently, to be undergone to the accreditation process. In conclusion, the results demonstrated an elevated robustness and reliable features of the "PCR E.g.s.s." able to perform a rapid diagnosis of E. granulosus in only "one step", hence, it is likely to avoid the sequencing step.


Assuntos
Echinococcus granulosus/classificação , Echinococcus granulosus/genética , Reação em Cadeia da Polimerase/métodos , Animais , Equinococose/classificação , Equinococose/genética , Genótipo , Itália , Epidemiologia Molecular/métodos , Reprodutibilidade dos Testes
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