RESUMO
Side streams from aquaculture production such as fish sludge poses ample opportunities for biological upcycling, as the sludge contains high amounts of nutrients, energy and valuable biochemicals, making it an ideal food for extractive species. Sludge has been proposed as a feed stock for polychaete production, which in turn can be utilized live in shrimp aquaculture or as an aquafeed ingredient. However, the biosafety of such value chains has not yet been addressed. We conducted an experiment exposing the polychaete Hediste diversicolor to aquaculture sludge spiked with four different fish pathogens (Mycobacterium salmoniphilum, Yersinia ruckeri, Infectious Pancreatic Necrosis (IPN) and Infectious Salmon Anaemia (ISA)) known to cause diseases in Atlantic salmon (Salmo salar L.). Moreover, we assessed whether heavy metals and other potentially hazardous elements present in fish sludge bioaccumulates in the polychaetes. Neither of the bacteria nor viruses could be detected in the polychaetes after 14 days of continuous exposure. Seven of the 15 elements we analysed showed bioaccumulation factors significantly below one, meaning biodilution, while the other eight did not differ from one, meaning no bioaccumulation. None of the elements showed a significant bioaccumulation. Further on, none of the heavy metals found in the polychaetes at the end of our experiment exceeded the EU regulatory maximum levels for fish feed ingredients. The current results suggest that a H. diversicolor can reared on aquaculture sludge, and aquaculture sludge may serve as feed stock for polychaete production without the product exceeding EU regulations for contaminants in animal feed.
Assuntos
Aquicultura , Poliquetos , Esgotos , Animais , Poliquetos/metabolismo , Bioacumulação , Metais Pesados/metabolismo , Metais Pesados/análise , Salmo salar/metabolismo , Salmão/metabolismoRESUMO
Aquaculture sludge (uneaten feed and faeces) is nutrient rich and has potential as feed for insects. The aim of this study was to investigate the transfer of chemical and biological contaminants, as well as nutrients, from aquaculture sludge to black soldier fly larvae. The larvae were reared on a sludge mixture made of different sludges collected from Norwegian freshwater salmonid facilities. The sludge was spiked with four common salmon pathogens: Infectious Pancreatic Necrosis Virus, Infectious Salmon Anemia virus, Yersinia ruckeri or Mycobacterium salmoniphilum. During the 15 days of growth on sludge, the black soldier fly larvae accumulated valuable nutrients including protein, fat, eicosapentaenoic acid, iron, manganese, zinc and selenium. The larvae also accumulated undesirable substances including cadmium, mercury, dioxins and polychlorinated biphenyls. The concentrations of dioxins exceeded the EU maximum level set for animal feed. None of the salmon pathogens that were spiked to the sludge were detected in the black soldier fly larvae. This study reports low risk of transfer of salmon pathogens from sludge to insect larvae, and showed that the transfer of heavy metals, minerals and metalloids are in accordance with earlier studies. The large variations in levels of heavy metals between batches of sludge can cause levels in BSF exceeding the EU maximum levels, and thus indicate a need for monitoring of the proposed value chain. The transfer of dioxins from sludge to insects, reported for the first time in this paper, would be of special interest for future research, with special focus on risk mitigation.
Assuntos
Dípteros , Nutrientes , Esgotos , Nutrientes/química , Esgotos/química , Animais , Perciformes , Linhagem Celular , Salmão , Resíduos , Ração Animal , LarvaRESUMO
A total of 47 fish sludge samples from commercial land-based Atlantic salmon (Salmo salar) farms in Norway were assessed for their nutrient composition, presence of various legacy contaminants and a wide spectrum of contaminants of emerging concern, veterinary medicines as well as selected salmonid pathogenic bacteria and virus. The aim was to document the levels of desirable and undesirable components in fish sludge in relation to a potential future use of sludge as invertebrate feed. The samples had variable, but relatively high protein and fat contents, indicating a high load of undigested feed in some of the sludge samples. Fatty acid analysis showed the presence of essential omega-3 fatty acids. In terms of undesirable substances, 43% and 84% of the sludge samples contained levels of arsenic and cadmium, respectively, which exceeded the EU Maximum Levels established for complete animal feed. The concentrations of copper, zinc, iron and aluminum were highly variable in the sludge samples. The concentrations of dioxins, sum PCB6, and chlorinated pesticides were all below the Maximum Levels for animal feed. Of the 18 per- and polyfluoroalkyl substances (PFAS) only one compound (L-PFOS) was present at measurable levels. None of the samples had detectable levels of veterinary medicines, salmonid virus or bacteria. Performing a suspect and non-target screening of the sludge samples identified 18 compounds, including four pharmaceuticals, plastic-related products and the UV filter benzophenone, warranting further investigations. Overall, the results from this study show that fish sludge is a nutrient-rich resource; however, undesirable substances, originating from the feed or from treatment of sludge may be present.
Assuntos
Salmo salar , Esgotos , Animais , Nutrientes/análise , Ração Animal/análise , AquiculturaRESUMO
A confirmatory method for the determination of polypeptide antibiotics (bacitracin, colistin, and polymyxin B) in muscle samples has been developed. Extraction is performed with acidified methanol, and a clean-up step by solid-phase extraction with polymeric cartridges is applied. Separation by ultra-high performance liquid chromatography (UHPLC) is carried out using a solid core C18 column and gradient elution with water/acetonitrile containing 0.2% formic acid. High-resolution mass spectrometry (HRMS) (Q-Orbitrap) detection using different working modes has proved to be highly advantageous in eliminating interfering signals from endogenous matrix components. The analytical method has been successfully validated according to Commission Regulation 2021/808/EU and is currently used in a public health laboratory involved in veterinary medicines residue surveillance activities.
Assuntos
Antibacterianos , Espectrometria de Massas em Tandem , Animais , Antibacterianos/análise , Espectrometria de Massas em Tandem/métodos , Cromatografia Líquida de Alta Pressão/métodos , Músculos/química , Peptídeos , Extração em Fase Sólida/métodosRESUMO
BACKGROUND: Aquaculture aims to reduce the environmental and climate footprints of feed production. Consequently, low trophic marine (LTM) resources such as blue mussels and kelp are potential candidates to be used as ingredients in salmon feed. It is relevant to study potential undesirables associated with their use, as well as assessing food safety by investigating their transfer from feed-to-fish. The marine biota is well known to contain relatively high levels of arsenic (As), which may be present in different organic forms depending on marine biota type and trophic position. Thus, it is important to not only obtain data on the concentrations of As, but also on the As species present in the raw materials, feed and farmed salmon when being fed novel LTM feed resources. METHODS: Atlantic salmon were fed experimental diets for 70 days. A total of nine diets were prepared: four diets containing up to 4 % fermented kelp, three diets containing up to 11 % blue mussel silage, and one diet containing 12 % blue mussel meal, in addition to a standard reference diet containing 25 % fish meal. Concentrations of As and As species in feeds, faeces, liver and fillet of Atlantic salmon were determined by inductively coupled plasma mass spectrometry (ICP-MS) and high-performance liquid chromatography coupled to ICP-MS (HPLC-ICP-MS), respectively. RESULTS: The use of kelp or blue mussel-based feed ingredients increased the concentration of total As, but maximum level as defined in Directive 2002/32 EC and amendments was not exceeded. The concentrations found in the experimental feeds ranged from 3.4 mg kg-1 to 4.6 mg kg-1 ww. Arsenic speciation in the feed varied based on the ingredient, with arsenobetaine dominating in all feed samples (36-60 % of the total As), while arsenosugars (5.2-8.9 % of the total As) were abundant in kelp-included feed. The intestinal uptake of total As ranged from 67 % to 83 %, but retention in fillet only ranged from 2 % to 22 % and in liver from 0.3 % to 0.6 %, depending on the marine source used. Fish fed feeds containing blue mussel showed higher intestinal uptake of total As when compared with fish fed feeds containing fermented kelp. Fish fed fermented kelp-based feeds had higher retained concentrations of total As when comparing with fish fed feeds containing blue mussel. Despite relatively high intestinal uptake of total As, inorganic and organic As, the retained concentrations of As did not reflect the same trend. CONCLUSION: Although the use of LTM feed ingredients increased the level of total As in this feeds, salmon reared on these diets did not show increased total As levels. The well-known toxic inorganic As forms were not detected in salmon muscle reared on LTM diets, and the non-toxic organic AsB was the dominant As species that was retained in salmon muscle, while the organic AsSug forms were not. This study shows that speciation analysis of the LTM resources provides valuable information of the feed-to-fish transfer of As, needed to assess the food safety of farmed Atlantic salmon reared on novel low trophic feeds.
Assuntos
Arsênio , Kelp , Mytilus edulis , Salmo salar , Animais , Alimentos Marinhos/análise , Ração Animal/análiseRESUMO
Microalgae and blue mussels are known to accumulate undesirable substances from the environment, including arsenic (As). Microalgae can biotransform inorganic As (iAs) to organoarsenic species, which can be transferred to blue mussels. Knowledge on As uptake, biotransformation, and trophic transfer is important with regards to feed and food safety since As species have varying toxicities. In the current work, experiments were conducted in two parts: (1) exposure of the microalgae Diacronema lutheri to 5 and 10 µg/L As(V) in seawater for 4 days, and (2) dietary As exposure where blue mussels (Mytilus edulis L.) were fed with D. lutheri exposed to 5 and 10 µg/L As(V), or by aquatic exposure to 5 µg/L As(V) in seawater, for a total of 25 days. The results showed that D. lutheri can take up As from seawater and transform it to methylated As species and arsenosugars (AsSug). However, exposure to 10 µg/L As(V) resulted in accumulation of iAs in D. lutheri and lower production of methylated As species, which may suggest that detoxification mechanisms were overwhelmed. Blue mussels exposed to As via the diet and seawater showed no accumulation of As. Use of linear mixed models revealed that the blue mussels were gradually losing As instead, which may be due to As concentration differences in the mussels' natural environment and the experimental setup. Both D. lutheri and blue mussels contained notable proportions of simple methylated As species and AsSug. Arsenobetaine (AB) was not detected in D. lutheri but present in minor fraction in mussels. The findings suggest that low-trophic marine organisms mainly contain methylated As species and AsSug. The use of low-trophic marine organisms as feed ingredients requires further studies since AsSug are regarded as potentially toxic, which may introduce new risks to feed and food safety.
Assuntos
Arsênio , Microalgas , Mytilus edulis , Mytilus , Poluentes Químicos da Água , Animais , Arsênio/toxicidade , Arsênio/análise , Mytilus edulis/metabolismo , Microalgas/metabolismo , Cadeia Alimentar , Organismos Aquáticos/metabolismo , Poluentes Químicos da Água/análise , Mytilus/metabolismoRESUMO
BACKGROUND: Blue mussels (Mytilus edulis L.) can accumulate undesirable substances, including the potentially toxic elements (PTEs) cadmium (Cd), mercury, (Hg), lead (Pb), arsenic (As) and As species. In this study, the levels of PTEs and As species were determined in samples of blue mussels to assess the influence of environmental and biological factors, and evaluate the potential risk associated with blue mussels in terms of food and feed safety. METHODOLOGY: Blue mussels were collected monthly from one location in Western Norway from February 2018 to December 2018, and from April 2019 to April 2020. Samples were analyzed for PTEs using inductively coupled plasma mass spectrometry (ICP-MS), and high-performance liquid chromatography (HPLC) coupled to ICP-MS. Temperature, salinity and fluorescence (chlorophyll a) were monitored in the seawater column by STD/CTD, to assess the potential influence of these environmental factors on the PTE levels in the mussels. RESULTS: The results showed seasonal variations in the PTEs, with somewhat higher concentrations in spring and winter months. Unusually high levels of total As (101.2 mg kg-1 dw) and inorganic As (53.6 mg kg-1 dw) were observed for some of the time points. The organic As species arsenobetaine was generally the major As species (17-82% of total As) in the mussels, but also simple methylated As species and arsenosugars were detected. Principal components analysis (PCA) did not show a consistent relationship between the environmental factors and the PTE concentrations, showing contrary results for some elements for the periods studied. The condition index (CI) could explain variations in element concentration with significant correlations for Cd (r = -0.67, p = 0.009) and Pb (r = -0.62, p = 0.02 in 2019/20 and r = -0.52, p = 0.02 in 2018), whereas the correlation between As and CI was not significant (r = 0.12 in 2018, and r = -0.06 in 2019/20). Higher concentrations of iAs and arsenosugars coincided with increased signals of chlorophyll a, suggesting that phytoplankton blooms could be a source of As in the blue mussels. CONCLUSION: To our knowledge, this is the first study of As species in blue mussels collected over a time period of two years, providing an insight into the natural variations of these chemical forms in mussels. In terms of mussel as food and future feed material, concentrations of Cd, Hg and Pb were below the maximum levels (MLs) established in the EU food and feed legislation. However, levels of As and iAs in mussels at some time points exceeded the MLs for As in the feed legislation, and the margin of exposure (MOE) was low if these mussels were for human consumption, highlighting the importance of determining the chemical forms of As in feed and food.
Assuntos
Arsênio , Mercúrio , Mytilus edulis , Poluentes Químicos da Água , Animais , Humanos , Arsênio/análise , Cádmio/análise , Mercúrio/análise , Clorofila A/análise , Chumbo/análise , Estações do Ano , Monitoramento Ambiental/métodos , Noruega , Poluentes Químicos da Água/análiseRESUMO
A responsible harvest of mesopelagic species as aquafeed ingredients has the potential to address the United Nations Sustainable Development Goal 14, which calls for sustainable use of marine resources. Prior to utilization, the levels of undesirable substances need to be examined, and earlier studies on mesopelagic species have reported on total arsenic (As) content. However, the total As content does not give a complete basis for risk assessment since As can occur in different chemical species with varying toxicity. In this work, As speciation was conducted in single-species samples of the five most abundant mesopelagic organisms in Norwegian fjords. In addition, As species were studied in mesopelagic mixed biomass and in the resulting oil and meal feed ingredients after lab-scale feed processing. Water-soluble As species were determined based on ion-exchange high-performance liquid chromatography coupled to inductively coupled plasma mass spectrometry (HPLC-ICP-MS). This was supplemented by extracting arsenolipids (AsLipids) and determining total As in this fraction. The non-toxic arsenobetaine (AB) was the dominant form in mesopelagic crustaceans and fish species, accounting for approximately 70% and 50% of total As, respectively. Other water-soluble species were present in minor fractions, including carcinogenic inorganic As, which, in most samples, was below limit of quantification. The fish species had a higher proportion of AsLipids, approximately 35% of total As, compared to crustaceans which contained 20% on average. The feed processing simulation revealed generally low levels of water-soluble As species besides AB, but considerable fractions of potentially toxic AsLipids were found in the biomass, and transferred to the mesopelagic meal and oil. This study is the first to report occurrence data of at least 12 As species in mesopelagic organisms, thereby providing valuable information for future risk assessments on the feasibility of harnessing mesopelagic biomass as feed ingredients.
Assuntos
Arsênio , Animais , Arsênio/análise , Cromatografia Líquida de Alta Pressão/métodos , Crustáceos , Peixes , Espectrometria de Massas/métodos , ÁguaRESUMO
Organoarsenic species in marine matrices have been studied for many years but knowledge gaps still exist. Most literature focuses on monitoring of arsenic (As) species using previously published methods based on anion- and cation-exchange high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS). These studies are often limited to few As species and/or only specific method performance characteristics are described. Most marine certified reference materials (CRMs) are only certified for arsenobetaine (AB) and dimethylarsinate (DMA), making it difficult to evaluate the accuracy of analytical methods for other organoarsenic species. To address these gaps, the main objective of this work was to develop and validate a method for speciation analysis of a broad range of organoarsenic species in marine matrices. Optimum extraction conditions were identified through a 27-3 fractional factorial design using blue mussel as test sample. The effects of sample weight, type and volume of extraction solution, addition of H2O2 to the extraction solution, extraction time and temperature, and use of ultrasonication were investigated. The highest As recoveries were obtained by using 0.2 g as sample weight, 5 mL of aqueous methanol (MeOH:H2O, 50% v/v) as extractant, extraction carried out at 90 °C for 30 min, and without ultrasonication. Anion- and cation-exchange HPLC-ICP-MS settings were subsequently optimized. The method detected a total of 33 known and unknown As species within a run time of 23 and 20 min for cation-exchange and anion-exchange, respectively. A single-laboratory validation was conducted using several marine CRMs: BCR 627 (tuna fish tissue), ERM-CE278k (mussel tissue), DORM-4 (fish protein), DOLT-5 (dogfish liver), SQID-1 (cuttlefish), TORT-3 (lobster hepatopancreas), and CRM 7405-b (hijiki seaweed). Method performance characteristics were evaluated based on selectivity, limits of detection and quantification, linearity, trueness, precision, and measurement uncertainty. This work proposes an extraction procedure which allowed satisfactory quantification of As species with low solvent and energy consumption, supporting "Green Chemistry" principles. The study also presents a new set of As speciation data, including methylated arsenic species and arsenosugars, in recently issued marine CRMs, which will be valuable for future speciation studies on As. This work is the first to report a total of 33 different As species in marine CRMs.
RESUMO
The increase in the global population demands more biomass from the ocean as future food and feed, and the mesopelagic species might contribute significantly. In the present study, we evaluated the food and feed safety of six of the most abundant mesopelagic species in Norwegian fjords. Trace elements (i.e., arsenic, cadmium, mercury, and lead), organic pollutants (i.e., dioxins, furans, dioxin-like polychlorinated biphenyls, and polybrominated flame-retardants), and potentially problematic lipid compounds (i.e., wax esters and erucic acid) were analyzed and compared to existing food and feed maximum levels and intake recommendations. Furthermore, contaminant loads in processed mesopelagic biomass (protein, oil, and fish meal) was estimated using worst-case scenarios to identify possible food and feed safety issues. While most undesirables were low considering European food legislation, we identified a few potential food safety issues regarding high levels of fluoride in Northern krill, wax esters in glacier lanternfish, and long-chain monounsaturated fatty acids in silvery lightfish. Our estimates in processed biomass indicated high levels of undesirable trace elements in the protein fraction, frequently exceeding the maximum levels for feed ingredients. However, in fish meal, almost no exceedances were seen. In the oil fraction, dioxins and furans were above the maximum levels, given for food and feed ingredients. The present study is crucial to enable an evaluation of the value of these species; however, more data is needed before proceeding with large-scale harvesting of mesopelagic biomass.