Toxinas Algales en el Mar Argentino: nuevos hallazgos, nuevos desafíos / Algal Toxins in the Argentine Sea: new findings, new challenges

Resumen Las floraciones de algas nocivas son un problema cada vez más frecuente a nivel mundial que ocasiona severos daños sobre la salud pública, pérdidas económicas en acuicultura, perjuicios al turismo y episodios de mortalidad de poblaciones naturales de peces, aves y mamíferos marinos. Las toxinas son producidas por el fitoplancton y se acumulan en moluscos bivalvos que se alimentan por filtración del agua siendo estos los principales vectores de intoxicación humana. En el Mar Argentino, se han reportado toxinas marinas de origen microalgal asociadas con cuatro síndromes de intoxicación por moluscos. Los síndromes más graves por su extensión, frecuencia, toxicidad y organismos afectados, son los originados por el dinoflagelado Alexandrium cate-nella responsable de la Intoxicación Paralizante por Moluscos la cual ha ocasionado numerosas muertes humanas. Seguidamente, la más leve, en cuanto a gravedad y frecuencia, ha sido la Intoxicación Diarreica por Moluscos. En contraste, el ácido domoico, conocido como toxina amnésica de moluscos, no ha producido hasta ahora intoxicaciones humanas. Recientemente, se amplió el rango de toxinas para la región al registrarse las toxinas y los dinoflagelados productores de la Intoxicación Azaspirácidos por Moluscos. Además, se han detectado las potencialmente tóxicas Yessotoxinas y Espirolidos, cuyos mecanismos de acción y toxicidad están siendo aún evaluados a nivel mundial. Estas toxinas emergentes para la región, representan un riesgo potencial para la salud e inconvenientes socioeconómicos por el cierre de los sitios de explotación de moluscos. Ciertamente presentan un nuevo desafío, pues la detección y cuantificación sólo puede realizarse por medio de métodos basados en HPLC - espectrometría de masas, lo cual dificulta el monitoreo en laboratorios regionales en el país. La herramienta clave de manejo es la prevención, a través de políticas, regulaciones y sistemas de monitoreo y control de cada grupo de toxinas. A través de estas mejoras, se anticipa que no sólo disminuirá el número de afectados por estas intoxicaciones, si no que se podrán realizar vedas más eficientes, asegurando un equilibrio que proteja tanto la salud pública como el desarrollo de la industria pesquera.

Abstract Harmful algal blooms are an increasingly common problem worldwide, causing severe damage to public health, economic losses in aquaculture, damage to tourism and mortality events of natural populations of fish, birds and marine mammals. The toxins are produced by phytoplankton and accumulated in bivalve molluscs that feed on water filtration, being these main vectors of human intoxication. In the Argentine Sea marine toxins of microalgal origin have been reported associated with four shellfish poisoning syn-dromes. The most serious due to their extension, frequency, toxicity and affected organisms are those caused by the dinoflagellate Alexandrium catenella responsible for the Paralytic shellfish poisoning that has caused numerous human deaths. Then, the mildest, in severity and frequency, is the Diarrhetic shellfish poisoning. In contrast, domoic acid, known as Amnesic shellfish toxin, has not produced human intoxications yet. Recently, toxins and dinoflagellate species causing Azaspiracid shellfish poisoning have been re-corded, expanding the range of toxins for the region. In addition, the potentially toxic Yessotoxins and Spirolides have been detected, whose mechanism of action and toxicity is still being evaluated worldwide. These emerging toxins represent a potential risk to public health and socioeconomic activities due to the eventual closure of mollusc exploitation sites. They certainly present a new challenge, since detection and quantification can only be carried out using methods based on HPLC - mass spectrometry, which makes monitor-ing in regional laboratories difficult. Prevention through policies, regulations, and monitoring and control systems of each toxin group is the key management tool. These preventive measures are expected to contribute to reducing the number of poisonings and to ap-plying more efficient fisheries closures, ensuring a balance that protects both public health and the development of the fishing industry.

Genomic prediction for disease resistance to Hepatopancreatic parvovirus and growth, carcass and quality traits in Banana shrimp Fenneropenaeus merguiensis.

Conventional genetic improvement of disease resistance in aquatic animal species involves challenge tests or using qPCR to quantify viral load that is costly, time-consuming and causing biosecurity concerns. Recent developments in high throughput next generation genome sequencing platforms such as genotyping by sequencing (GBS) have opened new possibilities for improving disease traits based on DNA information. The principal aim of this study was thus to examine potential application of genomic selection to improve resistance to hepatopancreatic parvovirus (HPV) in banana shrimp Fenneropenaeus merguiensis. Specifically, we used a total of 9472 single nucleotide polymorphisms (SNPs) developed de novo from GBS platforms to assess accuracy of genomic prediction for HPV resistance and growth, carcass and quality-related traits in this white shrimp species. Our multi-locus mixed model analysis showed moderate heritabilities for HPV resistance (h2 = 0.46) and other traits studied (0.10 to 0.55). Genetic correlations of HPV titre with growth and carcass traits, estimated using SNPs markers, were negative (i.e., favourable), suggesting that selection for improved growth and carcass traits may have increased HPV resistance (i.e., reduced HPV titre). More importantly, our gBLUP model demonstrated that the accuracy of gBLUP prediction was moderate for HPV disease resistance (0.46). The genomic prediction accuracy was somewhat greater for growth and carcass related traits especially for body weight (0.76) and meat or tail weight (0.77). On the other hand, the prediction accuracy was from 0.25 to 0.41 for quality traits (raw and cooked colour and flesh streaks). Collectively, it is concluded that there are prospects to apply genomic selection in the genetic improvement for increased disease resistance, carcass and quality-related traits in this population of banana shrimp F. merguiensis.

Assessment of some heavy metal accumulation and nutritional quality of shellfish with reference to human health and cancer risk assessment: a seafood safety approach.

This study was conducted to assess the proximate analysis (protein, carbohydrate, lipid, and moisture content) and concentration levels of metals (Zn, Cu, Cd, Pb, and Cr) in the muscles of selected shellfish (Portunus reticulatus, P. segnis, P. sanguinolentus, Scylla olivaceae, Penaeus monodon, and P. indicus) species. The concentration of metals showed significant difference (p > 0.05) among species. The detected concentrations of the analyzed heavy metals were below the daily intake and legal limits set by national and international standards. The THQs and CR index were calculated to evaluate the risk estimation of the metal contamination associated with the human health. The THQ values of all metals were below 1 in all species, indicated that there is no risk of adverse health effect, but the risk of elevated intakes of heavy metals adversely affecting food safety for the studied species. The CR index indicated that Cd and Pb caused the greatest cancer risk. The correlation and multivariate (principle component analysis) among metal concentration and nutritional quality were also evaluated. The carbohydrate and moisture showed the positive correlation (p > 0.05) with metals. The biochemical results of the present work clearly indicate that there was a significant difference in the muscles of shellfish. It was concluded that more effective controls should be focused on Cd and Pb to reduce pollution for quality and seafood safety concern.

A sensory and nutritional comparison of mussels (Mytilus sp.) produced in NW Iberia and in the Armona offshore production area (Algarve, Portugal).

A biometric, nutritional and sensory analysis of raw and cooked mussels comparing Mytilus sp. from the north-west coast of Portugal and Spain (Minho and Galicia, respectively) and the new offshore production site of Armona (Algarve, south Portugal) was carried out. In addition, multiple factorial analysis was performed to explore potential relationships between sensory attributes and nutritional content properties between the different mussels. Results showed that, at similar times of sale, biometrics of mussels from Armona and Vigo were similar and bigger than the remaining. Nonetheless, despite some similarities in proximate composition, mussels presented differences in lipid classes, fatty acid content and free amino acids profiles. These differences were not fully reflected in the sensory assessment by the panel, which were able to distinguish different production sites in raw specimens but displayed problems in discrimination these in cooked mussels. Some nutritional components were related to specific sensory sensations.

Polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (dl-PCBs) in fish, seafood products and fish oil in Spain.

A total of 84 samples of wild and farmed fish, cephalopods and fish oils for animal feeding, traded in Spain, were analysed for polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (dl-PCBs) in 2009-2012, by gas chromatography-ion trap tandem mass spectrometry (GC-MS-MS). The method was optimised for screening at moderate costs, allowing PCDD/Fs determination at 1 pg World Health Organization-toxic equivalent quantities (WHO-TEQ) g⁻¹ wet weight (w w) and dl-PCBs at 0.02 pg WHO-TEQ g⁻¹ w w. Concentrations in fish and cephalopods ranged from values below the limit of detection to 1.7 pg g⁻¹ WHO-TEQ sum PCDD/Fs and dl-PCBs, considered as safe with regard to EU legislation. Higher levels were found in cod livers (5.4-54.2) and fish oils (3.3-30.7), with one noncompliant sample in each group.

Survey of polycyclic aromatic hydrocarbons in marine products in Korea using GC/MS.

This study investigates polycyclic aromatic hydrocarbons (PAHs) in marine products on the Korean market. A total of 280 samples of fish (n = 100), shellfish (n = 80), cephalopod (n = 60) and crustacea (n = 40) were collected for analyses of PAHs (naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benz[a]anthracene, chrysene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, indeno[1,2,3-c,d]pyrene, dibenz[a,h]anthracene and benzo[g,h,i]perylene). The analytical procedure was based on the matrix solid-phase dispersion on Florisil cartridges and extraction with hexane/dichloromethane (3:1, v/v). The PAHs were determined by gas chromatography with mass spectrometric detection using selective ion monitoring. Average recoveries for all the PAHs studied were in the range 58-79%. The sum of 16 PAHs concentrations in fish, shellfish, cephalopod/crustacea were in the range 0.2-0.5, 1.2-1.6 and 0.8-1.9 µg/kg, respectively.

Nutritional quality and safety of cooked edible crab (Cancer pagurus).

Edible crab (Cancer pagurus) is one of the most important crustaceans consumed in Southern European countries, either as boiled or steamed cooked product. So far, the majority of studies assessing health benefits and risks associated to seafood consumption have been carried out in raw products, despite being generally cooked before consumption, and mostly in muscle tissue. Therefore, the aim of this study was to assess the effect of steaming and boiling on the chemical safety and nutritional value of C. pagurus caught in spring and summer. Generally, chemical and elemental composition of brown meat (tissue in the body cavity comprising mainly gonads and hepatopancreas) differed significantly from muscle (white meat in claws and legs). Additionally, the same tissue revealed differences in chemical and elemental composition of raw and cooked C. pagurus, likely due to water leaching. In contrast, few variations between seasons were observed. The results revealed that the consumption of cooked edible crab muscle should be promoted, whereas brown meat ingestion should be done parsimoniously.

Cadmium in shellfish: the British Columbia, Canada experience--a mini-review.

Over 10 years ago, research scientists in the federal department of Fisheries and Oceans Canada (DFO) were alerted to the presence of high levels of cadmium, a toxic metal, in the Pacific oyster (Crassostrea gigas) cultured in British Columbia (BC), Canada waters. This mini-review summarizes the most recent published studies on levels of cadmium in shellfish from the Pacific Northwest (BC and Washington State). Recent health risk assessments with respect to the ingestion of BC oysters are presented. The review concludes with a discussion on the ethical dilemma now facing the shellfish industry in the Pacific Northwest and provides recommendations for the culturing of shellfish without fear of high cadmium levels.

Effect of pyrophosphate and 4-hexylresorcinol pretreatment on quality of refrigerated white shrimp (Litopenaeus vannamei) kept under modified atmosphere packaging.

The effect of pretreatment with pyrophosphate and 4-hexylresorcinol in combination with modified atmosphere packaging (MAP) (80% CO(2), 10% O(2), 10% N(2), or 80% CO(2), 20% N(2)) on the quality of white shrimp during storage at 4 degrees C was investigated. Shrimp pretreated with 2% pyrophosphate and 0.25% 4-hexylresorcinol and stored under MAP showed the lower microbiological and chemical deteriorations as evidenced by delayed microbial growth as well as lower trimethylamine (TMA) and total volatile base nitrogen (TVB) production (P < 0.05). Additionally, the growth of coliforms was inhibited effectively. White shrimp pretreated with 4-hexylresorcinol had the lower melanosis throughout the storage compared with those without treatment (P < 0.05). This was associated with the lowered polyphenol oxidase (PPO) activity in shrimp treated with 4-hexylresorcinol. Therefore, the effective retardation of microbiological and chemical deterioration of white shrimp stored under MAP with the decrease in melanosis could be achieved by pretreatment of the shrimp with pyrophosphate and 4-hexylresorcinol. Furthermore, decapitation could be another means to lower the microbial load and melanosis in white shrimp, particularly those stored under MAP.

Mercury, cadmium and lead levels in samples of the main traded fish and shellfish species in Galicia, Spain.

The presence of mercury, cadmium and lead has been studied in raw and processed fish and shellfish of the main traded species in Galicia (Spain) from at least 40 commercial outlets between January 2002 and October 2003. The results show that, according to European legislation, the products on the market are generally "safe", although the high levels of mercury in swordfish and shark are notable, with 22% of samples (n = 37) over the limit. We also indicate the relatively important contribution to mercury ingestion of widely consumed species, such as tuna, with 25% of samples (n = 378) over 0.25 mg kg⁻¹. Cadmium was over the limit (1.0 mg kg⁻¹) in 15% of the tested cephalopods (n = 79) and 29% of Chlamys sp. bivalves (n = 17).

Monitoring programme on cadmium, lead and mercury in fish and seafood from Valencia, Spain: levels and estimated weekly intake.

The study was carried out to determine the current levels of mercury, cadmium, and lead in fish and seafood from the market of Comunitat Valenciana, Spain. Levels of total mercury ranged from 0.02 to 3.15 mg kg⁻¹ w.w. (average = 0.073 mg kg⁻¹ w.w.). Cadmium concentrations ranged from 0.003 to 0.66 mg kg⁻¹ w.w. (average = 0.27 mg kg⁻¹ w.w.) for seafood, and between 0.003 and 0.71 mg kg⁻¹ w.w. (average = 0.01 mg kg⁻¹ w.w.) for marine fish. Concerning lead, concentrations from 0.02 to 0.36 mg kg⁻¹ w.w (average = 0.04 mg kg⁻¹ w.w.) were found in fish, and from 0.02 to 1.02 mg kg⁻¹ w.w. in seafood (average = 0.147 mg kg⁻¹ w.w.). The levels found were, in general, lower than maximum levels proposed by European legislation. The estimated weekly intake of mercury, cadmium and lead through fish and seafood consumption was found to be about 43%, 10% and 3% of the established provisional tolerable weekly intake, respectively.

Postmortem biochemical behavior of giant squid (Dosidicus gigas) mantle muscle stored in ice and its relation with quality parameters.

Several freshness and spoilage indicators were monitored to characterize the postmortem biochemistry of giant squid (Dosidicus gigas) mantle muscle. Squid samples were obtained directly from the sea and kept at 0 degrees C during a 15-d storage period. Data at zero time were obtained from cryogenically frozen samples at time of capture. The adenosine 5'-triphosphate (ATP) degradation followed a different pattern as compared with that from fish species. ATP was almost completely depleted at 24-h postcatch from 6.54 to <1 micromol/g, while at the same time Hx was the predominant catabolite with a concentration of 4 mumol/g, reaching 6.85 micromol/g at day 15. K-value data followed a logarithmic pattern with time instead of a linear one, with no change after day 3, thus reducing its suitability as a freshness index. The coefficient Hx/AMP seems to be an adequate alternative for this purpose due to its constant increment with time. The high NH4Cl content in mantle muscle (461.3 +/- 24.5 mg of NH4(+)/100 g) derived from its physiological importance for the species compromises the use of the distillation step of the TVB-N analysis commonly used as a spoilage index. This fact explains why the initially high value of TVB-N detected in mantle muscle (243.7 mg N/100 g) did not correlate with the initial low TMA-N content (1.5 +/- 0.1 mg/100 g of muscle). The results suggested that under the experimental conditions the shelf life of squid exceeds 15 d.

Microbial population, physicochemical quality, and allergenicity of molluscs and shrimp treated with cobalt-60 gamma radiation.

Frozen molluscs (squid, octopuses, and cuttlefish) and crustaceans (shrimp) were irradiated using a cobalt-60 gamma source, at different doses, in order to investigate the effects of gamma radiation on their microbial population, organoleptic characteristics, lipid profile, and tropomyosin content. Irradiation of shrimp and squid with either 2.5 or 4.7 kGy reduced mesophilic bacteria contamination to low or nondetectable levels, respectively, whereas irradiation of octopus and cuttlefish with the same doses reduced the bacterial population. Irradiation treatment had no significant (P > 0.05) effect on the total lipid content and the major detected classes of polar and neutral lipids, whereas it significantly (P < 0.05) increased the contents of neutral lipids in octopus mantle and in shrimp muscle and cephalothorax samples. The total fatty acid content and the omega-3: omega-6 fatty acid ration was not affected. A dose-dependent significant (P < 0.05) decrease in the ratio of polyunsaturated fatty acids:saturated fatty acids was observed. With the increase in radiation dose, redness (a) and yellowness (b) values showed a variation, whereas the lightness (L) value was significantly (P < 0.05) decreased in mollusc mantles and shrimp muscle and increased in shrimp cephalothorax. The total of color changes ( delta E) increased (P < 0.05) as the dose increased. Significant (P < 0.05) changes in textural properties were observed with radiation treatment in octopus tentacles and in squid and cuttlefish mantle. The amount of tropomyosin, which is the major mollusc and crustacean allergen in the irradiated organisms, was reduced by gamma radiation, depending on the dose.

Inactivation by ionizing radiation of Salmonella enteritidis, Salmonella infantis, and Vibrio parahaemolyticus in oysters (Crassostrea brasiliana).

Irradiation is considered one of the most efficient technological processes for the reduction of microorganisms in food. It can be used to improve the safety of food products, and to extend their shelf lives. Oysters are considered one of the most important vehicles for pathogenic bacteria because of their feeding characteristics. The aim of this study was to evaluate the influence of a gamma radiation process on high levels of Salmonella Enteritidis, Salmonella Infantis, and Vibrio parahaemolyticus incorporated by oysters (Crassostrea brasiliana), as well as the effects of the process on the survival of the oysters and on their sensory attributes. The oysters were exposed to gamma radiation (60Co) in doses ranging from 0.5 to 3.0 kGy. A dose of 3.0 kGy was generally sufficient to reduce the level of Salmonella serotypes by 5 to 6 log10 units. A dose of 1.0 kGy was sufficient to produce a 6-log10 reduction in the level of V. parahaemolyticus. The highest irradiation dose did not kill the oysters or affect their sensory attributes. Hence, a dose of 3.0 kGy can be considered effective in inactivating Salmonella and V. parahaemolyticus in oysters without changing their odor, flavor, or appearance.

Volatile components associated with bacterial spoilage of tropical prawns.

Analysis of headspace volatiles by gas chromatography/mass spectrometry from king (Penaeus plebejus), banana (P. merguiensis), tiger (P. esculentus/semisulcatus) and greasy (Metapenaeus bennettae) prawns stored in ice or ice slurry, which is effectively an environment of low oxygen tension, indicated the presence of amines at the early stages of storage (less than 8 days) irrespective of the nature of the storage media. Esters were more prevalent in prawns stored on ice (normal oxygen conditions) at the latter stages of storage (more than 8 days) and were only produced by Pseudomonas fragi, whereas sulphides and amines occurred whether the predominant spoilage organism was Ps. fragi or Shewanella putrefaciens. The free amino acid profiles of banana and king prawns were high in arginine (12-14%) and low in cysteine (0.1-0.17%) and methionine (0.1-0.2%). Filter sterilised raw banana prawn broth inoculated with a total of 15 cultures of Ps. fragi and S. putrefaciens and incubated for two weeks at 5 degrees C, showed the presence of 17 major compounds in the headspace volatiles analysed using gas chromatography/mass spectrometry (GC/MS). These were mainly amines, sulphides, ketones and esters. Principal Component Analysis of the results for the comparative levels of the volatiles produced by pure cultures, inoculated into sterile prawn broth, indicated three subgroupings of the organisms; I, Ps. fragi from a particular geographic location; II, S. putrefaciens from another geographic location; and III, a mixture of Ps. fragi and S. putrefaciens from different geographic locations. The sensory impression created by the cultures was strongly related to the chemical profile as determined by GC/MS. Organisms, even within the same subgrouping classified as identical by the usual tests, produced a different range of volatiles in the same uniform substrate.

Recuperación e identificación de vibrio choloerae en mariscos provenientes de diferentes mercados de Lima Metropolitana y Callao / Identification and recuperation of vibrio cholerae en sellfish from Lima and Callao market

A fines de 1991, se inició en el Perú una epidemia de cólera de gran inagnitud, en cuanto a morbi-mortalidad y extensión geográfica, que invadió a 19 países produciendo 600,000 casos y más de 6000 muertes. Esta enfermedad ha sido relacionada con el consumo de numerosos productos de pesca, entre ellos los mariscos como vehículos de transmisión del vibrio cholerae. El método utilizado fue el reconocimiento por el ICMSF modificado. Se analizó 104 muestras de mariscos, obteniéndose 0.96 por ciento de positividad correspondiente a la muestra de marisco tipo almeja, comprobándose mediantes pruebas serológicas, pertenecia al serotipo inaba. Asimismo, se comprobó la localización del microorganismo en las valvas del marisco y no en el músculo. También se puso de manifiesto la efectividad del método mediante un segundo enriquecimiento en APA pH=8,6 que proporcionó una mayor selectividad.