Cystic echinococcosis (CE) is a neglected zoonotic disease that remains a public-health concern worldwide. However, poor surveillance and incomplete reporting still hinder the calculation of the CE burden. This study evaluated national hospital discharge records (HDRs) from 2015 to 2022 to update CE epidemiological situation and economic impact of human CE in Italy. We reviewed 4,293 HDRs with an echinococcosis diagnosis during 2015-2022. Data extracted included age, sex, residence, nationality, primary versus secondary diagnosis, treatment type, length of stay and discharge outcome. The average annual hospital prevalence was calculated per 100,000 inhabitants and direct hospital costs were derived from records of CE as the primary diagnosis. The 2015-2022 trend was assessed and compared to an extended data set (2001-2022). From 2015 to 2022, 2,663 cases were identified nationally, over half in the southern and insular regions. The national average annual hospital prevalence was 0.56 cases/100,000 inhabitants, peaking in the islands (1.22/100,000 inhabitants). Patients were predominantly adults (>90%) and divided into the following age categories: 25-44 (20.4%), 45-64 (30.2%) and ≥65 years (41.1%); pediatric cases were rare (<3%). The mean hospital stay for primary CE diagnosis was 10.6 days with elderly patients (>65 years) accounting for 70% of in-hospital deaths. National direct hospital costs amounted to €10,726,413, with 56% borne by the South and Islands (South 23%, Islands 33%). A downward trend in hospitalised CE cases was observed from 2015 (326 cases) to 2022 (165 cases), with a notable decline in 2020 linked to the COVID-19 pandemic. Analysis of the longer time period (2001-2022) revealed 1250 fewer cases. These findings confirm CE's heterogeneous distribution, with persistent hyperendemic foci in traditional pastoral areas driving disproportionate healthcare costs. Sustained One Health surveillance, targeted regional prevention and robust national reporting are critical to further reduce CE burden in Italy.

Echinococcus multilocularis and Echinococcus granulosus sensu lato are the causative agents of two major zoonotic diseases, alveolar echinococcosis and cystic echinococcosis, respectively, and are ranked among the four most important foodborne parasites worldwide. Although interest in food contamination by taeniid eggs is increasing, the estimation of the number of eggs present in food items is still lacking. Using E. multilocularis eggs isolated from fecal samples of experimentally infected red foxes, an average of 5261 (±1253) mitochondrial DNA copies per egg was estimated by digital PCR (dPCR). Based on this estimation, 47 taeniid DNA samples previously obtained from different food items and identified as positive for E. multilocularis, E. granulosus sensu stricto or Taenia species, were submitted to dPCR to estimate the number of eggs present. In 95.7% of the samples, the contamination was estimated to be caused by one to five eggs, with only one egg in 83.0% of the samples. As detection of such low-level contamination requires sensitive detection methods, a performance comparison between microscopy and molecular methods was conducted using 15 pellets obtained from lettuce wash residues spiked with 0 to 63 E. multilocularis eggs. Two operators performed blinded microscopy, followed by DNA extraction from the examined pellets for real-time PCR detection and then an estimation of the number of eggs by dPCR. A higher sensitivity was obtained with real-time PCR (88.5%) compared to microscopy (40%) in samples containing one to five eggs. The correlation between the number of eggs spiked into the samples and the estimated egg number was strong using dPCR (r = 0.99, p < 0.01), while only moderate with microscopy (r = 0.67, p < 0.05). Estimation of the number of taeniid eggs in food samples by dPCR provides the first quantitative basis for exposure assessment and is important for evaluating the risk of human foodborne infections, with future work needed to address egg viability.

IntroductionThe African swine fever (ASF) is a viral disease of significant socio-economic and animal health impact, commonly diagnosed using molecular techniques. The World Organization of Animal Health (WOAH) has recommended two highly sensitive and specific real time polymerase chain reaction (PCR) procedures (P1 qPCR and P2 qPCR) for the reliable and rapid detection of the ASF virus (ASFV). The present work aimed to standardize two third-generation PCR technologies by using the same WOAH-recommended primer/probe sets.MethodsWe developed two droplet digital PCR protocols ddPCRs and compared their analytical performances to the above-mentioned qPCRs. This involved testing serially diluted plasmid containing the vp72 gene sequence of the ASFV as the template. Clinical positive and negative samples were also analyzed to compare both PCR procedures.Results and discussionThe ddPCR assays demonstrated excellent linearity (R2 = 0.999) across a dynamic range from 104 to 1 copies/μl. The limits of detections (LOD) were 3.48 and 2.80 copies/μl and the limit of quantifications (LOQs) were ranging from 25 and 20 copies/μl for the P1 and the P2 procedures, respectively. The LOD values were comparable to those of qPCRs assays. The analysis of the clinical samples evidenced a strong agreement between the qPCR and the ddPCR, with bias values below 0.20, as determined by Bland-Altman analysis. The results of this study indicated that the ddPCR method can be adapted to existing validated qPCR protocols, enabling the quantification of low viral titer samples with similar analytical sensitivity. Moreover, the use of ddPCR allows for the absolute quantification without the need of a calibration curve, providing a reliable tool for ASFV diagnosis and outbreak management. It could also support global effort to control the spread of ASFV. Further validation in diverse matrices (e.g., feed and environmental swabs) could expand its applicability in the One Health frameworks.

Cystic and alveolar echinococcosis are severe zoonotic diseases characterized by long asymptomatic periods lasting months or years. Viable Echinococcus spp. eggs released into the environment through the feces of canids can infect humans through accidental ingestion via hand-to-mouth contact or consumption of contaminated food or water. Both Echinococcus multilocularis and Echinococcus granulosus sensu lato are considered as foodborne parasites. However, when considering possible pathways of human infection, it appears that food and waterborne related variables do not significantly increase the risk of infection. Providing evidence-based data for the presence of DNA and, potentially, eggs in fresh produce is crucial in understanding foodborne transmission of Echinococcus spp. to humans. Two multicenter and multicountry studies were conducted within the One Health EJP framework to estimate the proportion of lettuces and berries contaminated by E. multilocularis, E. granulosus sensu lato, and other taeniid DNAs from a total of 12 European countries, Tunisia and Pakistan. A total of 1117 lettuces, 71 others vegetables, 300 strawberries, 130 blueberries and 50 others berries samples were collected and analysed by washing, sequential sieving and real-time PCRs. E. multilocularis DNA was detected in 1.2 % (7/ 570) of lettuce samples tested from the seven European endemic countries (Denmark, France, Germany, Latvia, the Netherlands, Poland and Switzerland) and in 2 % (2/100) from Pakistan. E. granulosus sensu lato DNA was identified in 1.3 % of lettuces (9/695) collected in five European endemic countries (France, Italy, Latvia, Poland and Portugal) and in 12 % (9/75) and 4 % (4/100) from Tunisia and Pakistan, respectively. All E. granulosus sensu lato samples were identified as E. granulosus sensu stricto (20/22), except for two identified as E. canadensis (2/22) from Latvia and Pakistan. Regarding berries, E. multilocularis DNA was detected in 5.4 % (n = 11/202) of strawberries, 7.3 % (6/82) of blueberries from the seven European endemic countries and 56 % (14/25) of blueberries from Pakistan. High contamination rates of E. granulosus sensu stricto were found outside of Europe, with 12.0 % (3/25) in blueberries from Pakistan and 81.3 %. (13/16) in strawberries from Tunisia. The total contamination rate of all taeniid species DNA in lettuces (5.3 %; 59/1117), others vegetables (5.6 %; 4/71) and berries (12.1 %; 58/480) suggests that the transfer of taeniid eggs from carnivore feces to food is not uncommon. Although we assume that eggs are the source of the DNA detected in this study, the viability of such eggs is unknown. The detection of Echinococcus species in lettuces and berries suggests a potential risk of foodborne human infection. The relative contribution of this risk remains to be estimated. Further studies on food and environmental contamination are necessary to cover different epidemiological contexts and social habits, leading to a better understanding of human infections by Echinococcus spp. eggs.

Few studies have analysed the current status of cystic echinococcosis (CE) in Pakistan. The study aimed to molecularly characterise Echinococcus infections in humans in Khyber Pakhtunkhwa (KP), Pakistan. A total of 251 formalin-fixed, paraffin-embedded (FFPE) CE cysts, with the clinical data of patients, were collected from four major hospitals in Peshawar, KP, during 2007-2021. The genomic DNAs (gDNA) containing 106 samples were successfully extracted. A multiplex PCR and a real-time PCR assay were performed to detect Echinococcus species. All 106 samples were confirmed as E. granulosus sensu lato and identified as E. granulosus sensu stricto (s.s.). Further analysis by using SNP genotyping showed that the G1 (79.3 %) genotype was the most prevalent compared to G3 (20.7 %). The study pointed out that E. granulosus s.s. G1 genotype is the primary cause of cystic echinococcosis (CE) in Peshawar, highlighting sheep as a major transmission source. The findings underscore the persistent zoonotic risk due to human-livestock interactions over 14 years. The limited access to healthcare facilities for early diagnosis and treatment, poor awareness about the disease among the population, and traditional livestock management practices may be possibly considered as the major factors causing the persistence of human disease in this area. To reduce the disease's impact on human health and economic losses in livestock, further epidemiological studies are essential to support effective control programs against this zoonotic disease in Pakistan.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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).

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.

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.

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.

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.

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.