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Detection of Trace Elements in the Bivalve Ruditapes Decussatus

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Detection of Trace Elements in the Bivalve Ruditapes Decussatus Università degli Studi di Sassari SCUOLA DI DOTTORATO DI RICERCA Scienze Veterinarie Indirizzo Qualità e Sicurezza Alimentare Ciclo XXXI Detection of trace elements in the bivalve Ruditapes decussatus from Sardinian coastal lagoons: effects on food safety and pathological findings in target organs Dott. Giuseppe Esposito Coordinatore Prof.ssa Fiammetta Berlinguer Referente di Indirizzo Prof. Enrico P.L. De Santis Docente Tutor Dott. Domenico Meloni Docente co-Tutor Dott. Marino Prearo Dott.ssa Elisabetta Antuofermo Dott. Antonio Pais Anno accademico 2018‐2019 2 A mio nonno Ferruccio Giuseppe Esposito – “Detection of trace elements in the bivalve Ruditapes decussatus from Sardinian coastal lagoons: effects on food safety and pathological findings in target organs” - Tesi di Dottorato in Scienze Veterinarie, indirizzo Qualità e Sicurezza Alimentare. Università degli Studi di Sassari. 3 INDEX Abstract 7 Chapter I General introduction 1.1 An overview of aquaculture production 12 1.1.1 Production from capture fisheries 14 1.1.2 Marine capture production 14 1.1.3 Inland waters capture production 14 1.1.4 Production from aquaculture 15 1.1.5 Marine and coastal aquaculture 15 1.1.6 Inland aquaculture 16 1.2 Aquaculture production systems 17 1.2.1 Extensive aquaculture in the Mediterranean 18 1.2.2 Extensive aquaculture in Italy 19 1.2.3 Extensive aquaculture in Sardinia 20 1.3 Introduction to Bivalve Molluscs 21 1.3.1 Global production of Bivalve Molluscs 23 1.3.2 Bivalve production in Europe 24 1.3.3 Current status of Bivalves production in Italy 25 1.4 Controls on production and processing 27 1.4.1 Classification of production and relaying areas 29 1.4.2 Purification of Bivalve Molluscs 29 1.4.3 Monitoring of classified relaying and production areas 36 1.5 Bivalve contamination and their risk as vehicles of diseases 40 1.5.1 Bacterial infections 41 1.5.2 Viral infections 42 1.5.3 Parasitic protozoa 43 1.5.4 Marine phytoplankton 44 1.5.5 Chemical compounds 45 Giuseppe Esposito – “Detection of trace elements in the bivalve Ruditapes decussatus from Sardinian coastal lagoons: effects on food safety and pathological findings in target organs” - Tesi di Dottorato in Scienze Veterinarie, indirizzo Qualità e Sicurezza Alimentare. Università degli Studi di Sassari. 4 1.5.5.1 Inorganic compounds 47 1.5.5.2 Organic compounds 55 1.6 Bivalve Molluscs as biological indicator of water pollution 58 1.6.1 The Grooved carpet shell Ruditapes decussatus 61 1.6.2 The Shell 61 1.6.3 The Mantle 64 1.6.3.1 Histology of the mantle 65 1.6.4 The Gills 65 1.6.4.1 Histology of the gills 66 1.6.5 The Reproductive system 67 1.6.5.1 Histology of the gonads 68 1.6.6 The digestive system 69 1.6.6.1 Histology of the digestive system 71 1.6.7 The cardio-vascular system 73 1.6.7.1 Histology of the cardio-vascular system 74 1.6.8 The nervous system 74 1.7 References 75 Chapter II Detection of trace elements in the bivalve Ruditapes decussatus from Sardinian coastal lagoons: effects on food safety 2.1 Introduction 99 2.2 Materials and Methods 101 2.2.1 Study area 101 2.2.2 Lagoon characteristics 101 2.2.3 North western Sardinia 101 2.2.3.1 The Calich lagoon 102 2.2.4 North eastern Sardinia 104 2.2.4.1 The Porto Pozzo lagoon 104 2.2.4.2 The San Teodoro lagoon 105 2.2.5 Central western Sardinia 106 Giuseppe Esposito – “Detection of trace elements in the bivalve Ruditapes decussatus from Sardinian coastal lagoons: effects on food safety and pathological findings in target organs” - Tesi di Dottorato in Scienze Veterinarie, indirizzo Qualità e Sicurezza Alimentare. Università degli Studi di Sassari. 5 2.2.5.1 The Marceddì-San Giovanni lagoon 106 2.2.6 South Sardinia 108 2.2.6.1 The Santa Gilla lagoon 108 2.2.7 Sampling procedures 110 2.2.8 Morphometric measures 111 2.2.9 Detection of trace elements 111 2.3 Statistical analyses 114 2.4 Results and Discussion 114 2.4.1 Morphometric measurements and Condition Index 114 2.4.2 Trace elements concentration in Ruditapes decussatus 119 2.4.2.1 Non-essential trace elements 124 2.4.2.2 Cadmium, lead and mercury 124 2.4.2.3 Aluminium and arsenic 125 2.4.2.4 Silver and tin 128 2.4.2.5 Essential trace elements 129 2.4.2.6 Cobalt and chromium 129 2.4.2.7 Copper, iron and manganese 129 2.4.2.8 Nickel, selenium and zinc 131 2.4.2.9 Metals levels in clams from different countries 135 2.4.3 Conclusions 139 2.4.4 References 140 Chapter III Histological indices and inflammatory responses in target organs 3. Introduction 150 3.1 Materials and Methods 156 3.1.1 Description of the study area and sampling procedures 156 3.1.2 Necropsy 157 3.1.3 Histopathology 158 3.1.4 Histopathological indices 161 Giuseppe Esposito – “Detection of trace elements in the bivalve Ruditapes decussatus from Sardinian coastal lagoons: effects on food safety and pathological findings in target organs” - Tesi di Dottorato in Scienze Veterinarie, indirizzo Qualità e Sicurezza Alimentare. Università degli Studi di Sassari. 6 3.1.5 Statistical analyses 162 3.2 Results and Discussion 163 3.2.1 Morphometric measures 163 3.2.2 Calich lagoon 163 3.2.3 Porto Pozzo lagoon 164 3.2.4 San Teodoro lagoon 166 3.2.5 Santa Gilla lagoon 168 3.2.6 Digestive gland histopathology 170 3.2.7 Gills histopathology 172 3.2.8 Kidney histopathology 175 3.3 Conclusions 186 3.4 References 187 Giuseppe Esposito – “Detection of trace elements in the bivalve Ruditapes decussatus from Sardinian coastal lagoons: effects on food safety and pathological findings in target organs” - Tesi di Dottorato in Scienze Veterinarie, indirizzo Qualità e Sicurezza Alimentare. Università degli Studi di Sassari. 7 Abstract Coastal waters and lagoons are typical environments devoted to extensive aquaculture. In Italy, which annual production is nearly 133,731 tonnes, shellfish farming is the main extensive aquaculture activity with an average production of about 119,166 tonnes in 2016 and representing over 90% of total production from transitional environments. Shellfish farming, being practiced above all in these ecosystems, is by its nature a very fragile productive sector. These environments represent the most widespread model of transition systems in the Mediterranean, whose possible pollution status must be assessed both in terms of the health status of the ecosystem and of direct or indirect risk to human health. The anthropogenic contamination of water bodies with heavy metals via fertilizer, industrial sewage and urban wastewater has resulted in widespread problems in aquatic organisms, but also poses a risk to consumer health. Shellfish from coastal and estuarine environments bioaccumulate toxic metals in their tissues due to their ability to concentrate inorganic contaminants several orders of magnitude above ambient levels. Since biomonitoring by using bivalve molluscs is currently considered one of the most effective approaches for assessing the degree of pollution of coastal brackish environments, the aims of this PhD thesis were: a) to detect the content of trace elements in the Grooved carpet shell Ruditapes decussatus collected in Sardinian coastal lagoons (Western Mediterranean Sea, Italy) and their effects on food safety; b) to evaluate the pathological findings in its target organs. To this end, in the first experimental contribution of the PhD thesis, wild clams were collected from five different brackish areas of Sardinia devoted to extensive aquaculture. In detail, 400 clams were sampled during autumn 2016 (100 specimens from Calich, Porto Pozzo, San Teodoro and Santa Gilla lagoons). During springer 2017 instead, 100 Giuseppe Esposito – “Detection of trace elements in the bivalve Ruditapes decussatus from Sardinian coastal lagoons: effects on food safety and pathological findings in target organs” - Tesi di Dottorato in Scienze Veterinarie, indirizzo Qualità e Sicurezza Alimentare. Università degli Studi di Sassari. 8 clams were harvested from Calich, Marceddì, Porto Pozzo and Santa Gilla lagoons (400 in total). The concentration of 16 trace elements (Al, Ag, As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Se, Sn, Tl and Zn) in the tissues of the clams was quantified. On the other hand, for the year 2017 the concentration of inorganic arsenic (iAs) was also considered as the sum of AsIII and AsV. The legal limits set by European Regulations for cadmium (Cd), mercury (Hg) and lead (Pb) were never exceeded. However, in both years, unexpectedly high values for aluminium (Al) and iron (Fe) (mean 127 and 105; 139.4 and 97.4 mg kg⁻1 wet weight, for 2016 and 2017 respectively) were found in Santa Gilla lagoon, which is close to industrial settlements and had the highest values for most of the investigated chemical elements. Nevertheless, during spring elevated values for the same metals were recorded also in Marceddì lagoon (133.1 and 106.6 mg kg⁻1 w.w., Al and Fe respectively), characterized by intensive zootechnical and agricultural activities. In 2016, the highest values of the metalloid arsenic (As) were reported in San Teodoro and Porto Pozzo (12.8 and 9.6 mg kg⁻1 w.w., respectively) while in 2017 the major value was found for Marceddì (3.4 mg kg⁻1 w.w.) followed by Porto Pozzo and Santa Gilla lagoons (8.2 and 3.3 mg kg⁻1 w.w., respectively). As for the concentration of inorganic arsenic (%iAs/tAs) instead, the highest values were recorded in Marceddì and Santa Gilla (2.76 and 1.94 mg kg⁻1 w.w., respectively).
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