Intermediate hosts of Toxoplasma gondii and food safety:epidemiology, genetics and parasite survival in meat-producing animals in Italy

Toxoplasma gondii is a coccidian parasite with a global distribution. The definitive host is the cat (and other felids). All warm-blooded animals can act as intermediate hosts, including humans. Sexual reproduction (gametogony) takes place in the final host and oocysts are released in the environment, where they then sporulate to become infective. In intermediate hosts the cycle is extra-intestinal and results in the formation of tachyzoites and bradyzoites. Tachyzoites represent the invasive and proliferative stage and on entering a cell it multiplies asexually by endodyogeny. Bradyzoites within tissue cysts are the latent form. T. gondii is a food-borne parasite causing toxoplasmosis, which can occur in both animals and humans. Infection in humans is asymptomatic in more than 80% of cases in Europe and North-America. In the remaining cases patients present fever, cervical lymphadenopathy and other non-specific clinical signs. Nevertheless, toxoplasmosis is life threatening if it occurs in immunocompromised subjects. The main organs involved are brain (toxoplasmic encephalitis), heart (myocarditis), lungs (pulmonary toxoplasmosis), eyes, pancreas and parasite can be isolated from these tissues. Another aspect is congenital toxoplasmosis that may occur in pregnant women and the severity of the consequences depends on the stage of pregnancy when maternal infection occurs. Acute toxoplasmosis in developing foetuses may result in blindness, deformation, mental retardation or even death. The European Food Safety Authority (EFSA), in recent reports on zoonoses, highlighted that an increasing numbers of animals resulted infected with T. gondii in EU (reported by the European Member States for pigs, sheep, goats, hunted wild boar and hunted deer, in 2011 and 2012). In addition, high prevalence values have been detected in cats, cattle and dogs, as well as several other animal species, indicating the wide distribution of the parasite among different animal and wildlife species. The main route of transmission is consumption of food and water contaminated with sporulated oocysts. However, infection through the ingestion of meat contaminated with tissue cysts is frequent. Finally, although less frequent, other food products contaminated with tachyzoites such as milk, may also pose a risk. The importance of this parasite as a risk for human health was recently highlighted by EFSA’s opinion on modernization of meat inspection, where Toxoplasma gondii was identified as a relevant hazard to be addressed in revised meat inspection systems for pigs, sheep, goats, farmed wild boar and farmed deer (Call for proposals -GP/EFSA/BIOHAZ/2013/01). The risk of infection is more highly associated to animals reared outside, also in free-range or organic farms, where biohazard measure are less strict than in large scale, industrial farms. Here, animals are kept under strict biosecurity measures, including barriers, which inhibit access by cats, thus making soil contamination by oocysts nearly impossible. A growing demand by the consumer for organic products, coming from free-range livestock, in respect of animal-welfare, and the desire for the best quality of derived products, have all led to an increase in the farming of free-range animals. The risk of Toxoplasma gondii infection increases when animals have access to environment and the absence of data in Italy, together with need for in depth study of both the prevalence and genotypes of Toxoplasma gondii present in our country were the main reasons for the development of this thesis project. A total of 152 animals have been analyzed, including 21 free-range pigs (Suino Nero race), 24 transhumant Cornigliese sheep, 77 free-range chickens and 21 wild animals. Serology (on meat juice) and identification of T. gondii DNA through PCR was performed on all samples, except for wild animals (no serology). An in-vitro test was also applied with the aim to find an alternative and valid method to bioassay, actually the gold standard. Meat samples were digested and seeded onto Vero cells, checked every day and a RT-PCR protocol was used to determine an eventual increase in the amount of DNA, demonstrating the viability of the parasite. Several samples were alos genetically characterized using a PCR-RFLP protocol to define the major genotypes diffused in the geographical area studied. Within the context of a project promoted by Istituto Zooprofilattico of Pavia and Brescia (Italy), experimentally infected pigs were also analyzed. One of the aims was to verify if the production process of cured “Prosciutto di Parma” is able to kill the parasite. Our contribution included the digestion and seeding of homogenates on Vero cells and applying the Elisa test on meat juice. This thesis project has highlighted widespread diffusion of T. gondii in the geographical area taken into account. Pigs, sheep, chickens and wild animals showed high prevalence of infection. The data obtained with serology were 95.2%, 70.8%, 36.4%, respectively, indicating the spread of the parasite among numerous animal species. For wild animals, the average value of parasite infection determined through PCR was 44.8%. Meat juice serology appears to be a very useful, rapid and sensitive method for screening carcasses at slaughterhouse and for marketing “Toxo-free” meat. The results obtained on fresh pork meat (derived from experimentally infected pigs) before (on serum) and after (on meat juice) slaughter showed a good concordance. The free-range farming put in evidence a marked risk for meat-producing animals and as a consequence also for the consumer. Genotyping revealed the diffusion of Type-II and in a lower percentage of Type-III. In pigs is predominant the Type-II profile, while in wildlife is more diffused a Type-III and mixed profiles (mainly Type-II/III). The mixed genotypes (Type-II/III) could be explained by the presence of mixed infections. Free-range farming and the contact with wildlife could facilitate the spread of the parasite and the generation of new and atypical strains, with unknown consequences on human health. The curing process employed in this study appears to produce hams that do not pose a serious concern to human health and therefore could be marketed and consumed without significant health risk. Little is known about the diffusion and genotypes of T. gondii in wild animals; further studies on the way in which new and mixed genotypes may be introduced into the domestic cycle should be very interesting, also with the use of NGS techniques, more rapid and sensitive than PCR-RFLP. Furthermore wildlife can become a valuable indicator of environmental contamination with T. gondii oocysts. Other future perspectives regarding pigs include the expansion of the number of free-range animals and farms and for Cornigliese sheep the evaluation of other food products as raw milk and cheeses. It should be interesting to proceed with the validation of an ELISA test for infection in chickens, using both serum and meat juice on a larger number of animals and the same should be done also for wildlife (at the moment no ELISA tests are available and MAT is the reference method for them). Results related to Parma ham do not suggest a concerning risk for consumers. However, further studies are needed to complete the risk assessment and the analysis of other products cured using technological processes other than those investigated in the present study. For example, it could be interesting to analyze products such as salami, produced with pig meat all over the Italian country, with very different recipes, also in domestic and rural contexts, characterized by a very short period of curing (1 to 6 months). Toxoplasma gondii is one of the most diffuse food-borne parasites globally. Public health safety, improved animal production and protection of endangered livestock species are all important goals of research into reliable diagnostic tools for this infection. Future studies into the epidemiology, parasite survival and genotypes of T. gondii in meat producing animals should continue to be a research priority.

Work-related health problems among resident immigrant workers in Italy and Spain

Background: in both Spain and Italy the number of immigrants has strongly increased in the last 20 years, currently representing more than the 10% of workforce in each country. The segregation of immigrants into unskilled or risky jobs brings negative consequences for their health. The objective of this study is to compare prevalence of work-related health problems between immigrants and native workers in Italy and Spain. Methods: data come from the Italian Labour Force Survey (n=65 779) and Spanish Working Conditions Survey (n=11 019), both conducted in 2007. We analyzed merged datasets to evaluate whether interviewees, both natives and migrants, judge their health being affected by their work conditions and, if so, which specific diseases. For migrants, we considered those coming from countries with a value of the Human Development Index lower than 0.85. Logistic regression models were used, including gender, age, and education as adjusting factors. Results: migrants reported skin diseases (Mantel-Haenszel pooled OR=1.49; 95%CI: 0.59-3.74) and musculoskeletal problems among those employed in agricultural sector (Mantel-Haenszel pooled OR=1.16; 95%CI: 0.69-1.96) more frequently than natives; country-specific analysis showed higher risks of musculoskeletal problems among migrants compared to the non-migrant population in Italy (OR=1.17; 95% CI: 0.48-1.59) and of respiratory problems in Spain (OR=2.02; 95%CI: 1.02-4.0). In both countries the risk of psychological stress was predominant among national workers. Conclusions: this collaborative study allows to strength the evidence concerning the health of migrant workers in Southern European countries.

Nosocomios higienistas: el caso Florence Nightingale

Los impresionantes cambios surgidos tras el inicio de la Revolución Industrial ayudaron a que de manera reiterada se buscaran soluciones a los diferentes problemas que se sucedían, uno tras de otro, como resultado del hacinamiento y las modificaciones sociopolíticas y económicas. Los avances en el campo de la ciencia, y en particular en las ciencias de la salud, fueron innumerables, estableciéndose en la sociedad una preocupación marcada por solucionar los problemas de las clases más desfavorecidas. En el campo de la sanidad, progresivamente se fueron dejando atrás posturas ancladas en los modelos basados en los «miasmas», para pasar a fundamentarse en el modelo microbiano de la mano de Pasteur y Lister. Estos descubrimientos fueron trasladados a la arquitectura progresivamente, dando paso a edificios que buscaban mantener unas condiciones higiénicas adecuadas que exiliaran las enfermedades del interior de los «paramentos de los hospitales». Ejemplo de esta lucha fue la llevada a cabo por la enfermera Florence Nightingale, que gracias a la observación y a sus conocimientos adquiridos tanto en el Instituto de Diaconisas de Kaiserswerth en Alemania como en la Maison de la Providence de las Hermanas de la Caridad de París, logró establecer una serie de pautas que aplicó al establecimiento que gestionó, el Hospital Scutari, durante la Guerra de Crimea.

The Early Miocene “Bisciaro volcaniclastic event” (northern Apennines, Italy): a key study for the geodynamic evolution of the central-western Mediterranean

The Early Miocene Bisciaro Fm., a marly limestone succession cropping out widely in the Umbria–Romagna–Marche Apennines, is characterized by a high amount of volcaniclastic content, characterizing this unit as a peculiar event of the Adria Plate margin. Because of this volcaniclastic event, also recognizable in different sectors of the central-western Mediterranean chains, this formation is proposed as a “marker” for the geodynamic evolution of the area. In the Bisciaro Fm., the volcaniclastic supply starts with the “Raffaello” bed (Earliest Aquitanian) that marks the base of the formation and ends in the lower portion of the Schlier Fm. (Late Burdigalian–Langhian p.p.). Forty-one studied successions allowed the recognition of three main petrofacies: (1) Pyroclastic Deposits (volcanic materials more than 90 %) including the sub-petrofacies 1A, Vitroclastic/crystallo-vitroclastic tuffs; 1B, Bentonitic deposits; and 1C, Ocraceous and blackish layers; (2) Resedimented Syn-Eruptive Volcanogenic Deposits (volcanic material 30–90 %) including the sub-petrofacies 2A, High-density volcanogenic turbidites; 2B, Low-density volcanogenic turbidites; 2C, Crystal-rich volcanogenic deposits; and 2D, Glauconitic-rich volcaniclastites; (3) Mixing of Volcaniclastic Sediments with Marine Deposits (volcanic material 5–30 %, mixed with marine sediments: marls, calcareous marls, and marly limestones). Coeval volcaniclastic deposits recognizable in different tectonic units of the Apennines, Maghrebian, and Betic Chains show petrofacies and chemical–geochemical features related to a similar calc-alkaline magmatism. The characterization of this event led to the hypothesis of a co-genetic relationship between volcanic activity centres (primary volcanic systems) and depositional basins (depositional processes) in the Early Miocene palaeogeographic and palaeotectonic evolution of the central-western Mediterranean region. The results support the proposal of a geodynamic model of this area that considers previously proposed interpretations.

Marine reptiles from the Triassic of the Tre Venezie Area, Northeastern Italy / Olivier Rieppel --, Fabio Marco Dalla Vecchia --.

n.s. no.44(2001)

Rome, Italy, 1891 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Pianta di Roma, col patrocinio del comune di Roma. It was published by editore Loreto Pasqualucci [per] dall' Istituto cartografico italiano in 1891. Scale 1:6,000. Map in Italian.The image inside the map neatline is georeferenced to the surface of the earth and fit to the European Datum 1950, Universal Transverse Mercator (UTM) Zone 33N projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, railroads and stations, drainage, built-up areas and selected buildings, walls, gates, fortification, ground cover, and more. Relief shown by shading. Includes also index and inset: Receniti di Roma nei successivi periodi della sua storia.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

Brescia, Italy, 1769 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Plan de Brescia. It was published by Desaint in 1769. Scale [ca. 1:7,448]. Covers a portion of Brescia, Italy. Map in French.The image inside the map neatline is georeferenced to the surface of the earth and fit to the European Datum 1950, Universal Transverse Mercator (UTM) Zone 32N projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, drainage, built-up areas and selected buildings, fortifications, ground cover, and more. Relief shown by hachures. Includes index.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

Padova, Italy, 1840 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Pianta della città di Padova, Vincenzo Voltolino, dis. e inc. It was published by V. Voltolino in 1840. Scale [ca. 1:11,000]. Covers the Podova region, Italy. Map in Italian.The image inside the map neatline is georeferenced to the surface of the earth and fit to the European Datum 1950, Universal Transverse Mercator (UTM) Zone 32N projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, drainage, built-up areas and selected buildings, fortifications, ground cover, and more. Includes views of buildings and index.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

Palermo, Italy, 1891 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Nuova pianta della città di Palermo. It was published by Carlo Clausen, editore in 1891. Scale 1:10,000. Covers a portion of Palermo, Italy. Map in Italian.The image inside the map neatline is georeferenced to the surface of the earth and fit to the European Datum 1950, Universal Transverse Mercator (UTM) Zone 33N projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, railroads and stations, drainage, built-up areas, selected buildings, parks, ground cover, shoreline features, and more. Includes index. Relief shown by shading.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

Parma, Italy, 1769 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Parma. It was published by Desaint in 1769. Scale [ca. 1:9,868]. Covers a portion of Parma, Italy. Map in Italian.The image inside the map neatline is georeferenced to the surface of the earth and fit to the European Datum 1950, Universal Transverse Mercator (UTM) Zone 32N projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, drainage, built-up areas and selected buildings, fortifications, ground cover, and more. Includes index.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

Piacenza, Italy, 1769 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Plan de la ville de Plaisance. It was published by Desaint in 1769. Scale [ca. 1:11,409]. Covers a portion of Piacenza, Italy. Map in French.The image inside the map neatline is georeferenced to the surface of the earth and fit to the European Datum 1950, Universal Transverse Mercator (UTM) Zone 32N projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, drainage, built-up areas and selected buildings, fortifications, ground cover, and more. Includes index.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

Genoa, Italy, 1910 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Nuova pianta topografica della città e del porto di Genova : secondo gli ultimi lavori stradali con il percorso dei trams. It was published by A. P. Editore in 1910. Scale [ca. 1:7,000]. Covers Genoa, Italy. Map in Italian.The image inside the map neatline is georeferenced to the surface of the earth and fit to the European Datum 1950, Universal Transverse Mercator (UTM) Zone 32N projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, railroads and stations, street railways, drainage, built-up areas and selected buildings, fortification, wharves, docks, gardens, and more. Relief shown by hachures.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

Milan, Italy, 1778 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Pianta della città di Milano, Arcan. Lavelli Ing. ... delir. Ang. Pestagalli F.Q. sculp. It was published in 1778. Scale [ca. 1:7,000]. Covers Milan, Italy. Map in Italian.The image inside the map neatline is georeferenced to the surface of the earth and fit to the European Datum 1950, Universal Transverse Mercator (UTM) Zone 32N projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, drainage, built-up areas and selected buildings, fortification, ground cover, parks, and more.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.