Preliminary data on the occurrence of PAH and Anisakis spp. in Moroccan anchovies: Environmental parasitology and human health risk.

This survey focuses on an environmental parasitology study by investigating Polycyclic Aromatic Hydrocarbons (PAH) bioaccumulation and the occurrence of Anisakis spp. on common anchovies collected from Moroccan coasts over 9 months through chemical (PAH) and Biological (Biometrics and parasitic) analysis. Obtained results were statistically analyzed and human health risks from anchovies consumption were assessed. The results obtained highlighted the good biological and chemical status of this fish in all stations of Morocco's coasts. Anisakis spp. was present in only four stations in the Atlantic fringe (maximum prevalence 22.22 %). Results have shown non-significant differences for tissue nature (Muscle, viscera) or spatial variation and were within a low range of concentrations well below the European Commission standards. Results have shown no serious harmful risk for humans from this fish consumption (ILCR and HI), and statistical analysis had shown positive correlations between prevalence and Chr, prevalence and sex-ratio, and prevalence and weight.

Primer reporte en Cuba de Anisakis spp. en muestras de pescado en conserva / First report in Cuba of Anisakis spp. in samples of canned fish

Introducción: Las especies de Anisakis representan uno de los grupos parasitarios patógenos transmitidos por alimentos más importantes en relación con sus capacidades zoonóticas y pueden producir una enfermedad emergente llamada anisakiosis. En Cuba, no existen reportes de la enfermedad como tampoco publicaciones sobre la detección del parásito en muestras de productos pesqueros. Objetivo: Identificar larvas de Anisakis spp. en muestras de productos pesqueros. Métodos: Se realizó un estudio para la búsqueda de larvas de Anisakis spp. en muestras de productos pesqueros en conserva recibidos en el Laboratorio de Parasitología de los alimentos del Instituto Nacional de Higiene, Epidemiología y Microbiología de Cuba. Se recibieron 96 muestras de dos marcas diferentes. A cada marca se le asignó las letras A o B para proteger la identidad del fabricante. Las larvas recuperadas fueron previamente aclaradas con lactofenol e identificadas morfológicamente por microscopía. Se utilizaron las claves de identificación para larvas de Anisakis spp. descritas en la literatura. Resultados: El 38,5 por ciento del total de muestras de pescado en conserva analizadas estuvieron parasitadas con larvas de Anisakis spp., con una frecuencia media de 1,2 larvas por cada submuestra. Las larvas fueron clasificadas de tercer estadio y de tipo I. Conclusiones: Por primera vez se identificó en Cuba larvas de Anisakis spp. en muestras de pescado en conserva y se proporcionó datos de interés a las autoridades sanitarias para el fortalecimiento de políticas públicas relacionadas con la calidad e inocuidad de los alimentos(AU)

Introduction: Anisakis species represent one of the most important foodborne pathogenic parasitic groups in relation to their zoonotic capabilities, and can produce an emerging disease called anisakiosis. In Cuba, there are no reports of the disease and there are no publications on the detection of the parasite in samples of fishery products. Objective: To identify Anisakis spp. larvae in samples of fishery products. Methods: A study was conducted to search for Anisakis spp. larvae in samples of canned fish products received at the Laboratory of Food Parasitology of the National Institute of Hygiene, Epidemiology and Microbiology of Cuba. Ninety-six samples of two different brands were received. Each brand was assigned letter A or B to protect the identity of the manufacturer. The larvae recovered were previously rinsed with lactophenol and identified morphologically by microscopy. The identification keys for Anisakis spp. larvae described in the literature were used. Results: 38.5 percent of the total canned fish samples analyzed were parasitized with Anisakis spp. larvae, with a mean frequency of 1.2 larvae per subsample. The larvae were classified as third instar and type I. Conclusions: For the first time in Cuba, Anisakis spp. larvae were identified in canned fish samples and provided data of interest to health authorities to strengthen public policies related to food quality and safety(AU)

Anisakidosis diagnosticada tras la expulsión de vermes en deposiciones: forma poco frecuente de diagnóstico en humanos / Anisakidosis diagnosed after expulsion of worms in stools: a rare form of diagnosis in humans

Resumen La anisakidosis es una zoonosis parasitaria accidental y cosmopolita de los seres humanos, siendo su hospedero definitivo los mamíferos marinos como lobos de mar, focas y delfines, entre otros. El ser humano se infecta por el consumo en estado larvario del nemátodo de la familia Anisakidae presentes en pescados y mariscos crudos como la merluza o el congrio. Los helmintos se ubican preferentemente en el tubo digestivo alto, observándose la regurgitación del parásito a la boca o evidenciándose durante la realización de una endoscopía digestiva alta. En forma infrecuente las larvas pueden migrar al peritoneo o seguir su paso por el intestino delgado y colon, siendo inhabitual la expulsión por las deposiciones. Presentamos el caso de una expulsión dos larvas L3 de la familia Anisakidae en deposiciones.

Abstract Anisakidosis is an accidental and cosmopolitan parasitic zoonosis of human beings, its definitive host being marine mammals such as sea lions, seals and dolphins, among others. Humans are infested by consumption in the larval stage of the nematode of Anisakis present in raw fish and shellfish such as hake or conger eel. The infestation is preferably located in the upper digestive tract, observing the regurgitation of the parasite to the mouth or becoming evident during an upper gastrointestinal endoscopy. In rare cases, the larvae can migrate to the peritoneum or continue their passage through the small intestine and colon, with expulsion in stools being unusual. We present a case of expulsion of two larvae of the Anisakidae family in feces.

Anisakid and Raphidascaridid parasites in Trachurus trachurus: infection drivers and possible effects on the host's condition.

This study investigated the distribution of nematode larvae of Anisakidae and Raphidascarididae (genera Anisakis and Hysterothylacium) in Trachurus trachurus (Linnaeus, 1758) in the Ligurian and central-northern Tyrrhenian Seas. The relationship between the number of parasites and the length and weight parameters of the fish was assessed, and the possible effect of the parasites on the condition factor was evaluated. A total of 190 T. trachurus specimens were collected in July 2019. Parasites were found in 70 individuals. A total of 161 visible larvae were collected in the viscera. Morphological analysis revealed the presence of Anisakis spp. in 55 fish and Hysterothylacium spp. in 15 fish, while 5 fish showed coinfection with both genera. The specimens subjected to PCR (n = 67) showed that 85% of the Anisakis larvae analyzed belonged to the species A. pegreffii, while the remaining 15% belonged to hybrids of A. pegreffii-A. simplex (s.s.). A total of 58% (n = 7) of the Hysterothylacium larvae analyzed belonged to the species H. fabri, while 42% belonged to the species H. aduncum. Our results support the hypothesis that infection with these parasites does not affect the condition of the fish host analyzed, and that body size and depth are major drivers in determining infection levels with Anisakid and Raphidascaridid nematodes.

Molecular and evolutionary basis for survival, its failure, and virulence factors of the zoonotic nematode Anisakis pegreffii.

Parasitism is a highly successful life strategy and a driving force in genetic diversity that has evolved many times over. Accidental infections of non-targeted hosts represent an opportunity for lateral host switches and parasite niche expansion. However, if directed toward organisms that are phylogenetically distant from parasite's natural host, such as humans, it may present a dead-end environment where the parasite fails to mature or is even killed by host immunity. One example are nematodes of Anisakidae family, genus Anisakis, that through evolution have lost the ability to propagate in terrestrial hosts, but can survive for a limited time in humans causing anisakiasis. To scrutinize versatility of Anisakis to infect an evolutionary-distant host, we performed transcriptomic profiling of larvae successfully migrating through the rat, a representative model of accidental human infection and compared it to that of larvae infecting an evolutionary-familiar, paratenic host (fish). In a homeothermic accidental host Anisakis upregulated ribosome-related genes, cell division, cuticle constituents, oxidative phosphorylation, in an unsuccessful attempt to molt to the next stage. In contrast, in the paratenic poikilothermic host where metabolic pathways were moderately upregulated or silenced, larvae prepared for dormancy by triggering autophagy and longevity pathways. Identified differences and the modelling of handful of shared transcripts, provide the first insights into evolution of larval nematode virulence, warranting their further investigation as potential drug therapy targets.

Anisakis spp. in toothed and baleen whales from Japanese waters with notes on their potential role as biological tags.

In this study, Anisakis nematodes isolated from toothed and baleen whales from localities around Japan were molecularly (PCR-RFLP) identified. In Wakayama, common bottlenose dolphins (Tursiops truncatus) were infected with A. simplex sensu stricto (s.s.), A. typica and A. pegreffii, while A. typica was the only species found in pantropical spotted dolphin (Stenella attenuata) and striped dolphin (S. coeruleoalba). Offshore common minke whales (Balaenoptera acutorostrata) and sei whales (B. borealis) were almost exclusively infected with A. simplex s.s.. However, in common minke whales from two Hokkaido localities, mature worms mostly consisted of A. simplex s.s. in some individuals and of A. pegreffii in others, but immature worms were mainly A. simplex s.s.. Gross and histopathological examination on gastric mucosa attached by anisakids resulted in mild and superficial reactions by the two baleen whale species in contrast to severe inflammatory reaction associated with ulcer formations by common bottlenose dolphin. Host specificity and adaptability of Anisakis spp. in these baleen and toothed whales were discussed from the points of view of adult worm size, worm population and pathological reactions by hosts. Interestingly, most of the common minke whales predominantly harboring mature A. pegreffii adults belonged to the Yellow Sea - East China Sea stock (J stock), which migrates through the Sea of Japan, whereas most of those mainly parasitized by mature A. simplex s.s. adults were from the Okhotsk Sea - West Pacific stock (O stock), mostly inhabiting the Pacific side, suggesting that these sibling species may have utility as biological tags to differentiate whale stocks. These results represent the first definitive host records for A. pegreffi in the Northwestern Pacific Ocean.

Anisakis spp. Larvae in Deboned, in-Oil Fillets Made of Anchovies (Engraulis encrasicolus) and Sardines (Sardina pilchardus) Sold in EU Retailers.

Sardina pilchardus and Engraulis encrasicolus are considered the principal target species for commercial fishing in Europe and are widely consumed as semipreserved products. Although they are considered shelf-stable products, if treatment is not correctly applied, their consumption may represent a public health risk in regard to anisakiasis and allergic reactions. Little is known about the prevalence of Anisakis spp. in ripened products. This study aimed to evaluate the presence of Anisakis spp. larvae in deboned, in-oil anchovy and sardine fillets marketed in the EU to assess the influence of processing techniques on the prevalence of larvae. Ninety semipreserved anchovy and sardine products deriving from the Mediterranean Sea or Atlantic Ocean were collected from different EU retailers and examined using chloropeptic digestion to evaluate the presence of larvae and identify them. Thirty nonviable Anisakid larvae-A. pegreffii (30%) and A. simplex (70%)-were found. The frequency of larvae was higher in anchovies (28.8%). The low frequency of parasites found proved that processing technologies can influence the presence of larvae in final products, but it is important that visual inspection is performed only by trained people. The sources of raw materials should be considered in the production flow chart.

Occurrence of larval anisakids in horse mackerel (Trachurus trachurus) caught in Portuguese waters.

The third-stage larvae (L3) of the Anisakidae family are parasitic nematodes with zoonotic impact and are frequently encountered in the organs and musculature of various fish intended for human consumption. Since Anisakis simplex (s.s.) and A. pegreffii are the major aetiological agents of human disease, this study aims to combine the morphological and molecular data on the recovered anisakid larvae to contribute to a simplified morphological distinction of those species and conducted a survey of anisakid larvae infection in horse mackerel (Trachurus trachurus). Here, 116 horse mackerel caught in Portuguese waters were analysed for the presence of L3 of anisakids, and 3148 larvae were collected, of which only 30% were retrieved during visual inspection. As such, visual inspection does not appear to be very effective in anisakid detection. A prevalence of 84.5% of infected fish was found, and the mean intensity and mean abundance were 32.1 and 27.1 parasites per fish, respectively. The morphological and molecular analyses of 196 L3 randomly chosen from the total sample of parasites demonstrated the presence of L3 of mostly Anisakis spp., with only one L3 of Hysterothylacium aduncum. Relative frequencies of 62.9% for A. pegreffii and 37.1% for A. simplex (s.s.) were obtained. The morphometry differences between these two sibling species were evaluated, and the results demonstrated significant differences between the length of the ventriculus and the length of the oesophagus. Precisely, A. simplex (s.s.) has a longer oesophagus and ventriculus than A. pegreffii. As such, these differences may be used to distinguish the two species through morphological analysis.

Mass Spectrometry Based-Proteomic Analysis of Anisakis spp.: A Preliminary Study towards a New Diagnostic Tool.

Anisakiasis is nowadays a well-known infection, mainly caused by the accidental ingestion of Anisakis larvae, following the consumption of raw or undercooked fishes and cephalopods. Due to the similarity of symptoms with those of common gastrointestinal disorders, this infection is often underestimated, and the need for new specific diagnostic tools is becoming crucial. Given the remarkable impact that MALDI-TOF MS biotyping had in the last decade in clinical routine practice for the recognition of bacterial and fungi strains, a similar scenario could be foreseen for the identification of parasites, such as nematodes. In this work, a MALDI-TOF MS profiling of Anisakis proteome was pursued with a view to constructing a first spectral library for the diagnosis of Anisakis infections. At the same time, a shotgun proteomics approach by LC-ESI-MS/MS was performed on the two main fractions obtained from protein extraction, to evaluate the protein species enriched by the protocol. A set of MALDI-TOF MS signals associated with proteins originating in the ribosomal fraction of the nematode extract was selected as a potential diagnostic tool for the identification of Anisakis spp.

Anisakis spp. in fishery products from Japanese waters: Updated insights on host prevalence and human infection risk factors.

The nematodes of the genus Anisakis are among the most relevant parasitic hazards in fishery products since they are responsible for human infection and allergy cases. In a food safety and epidemiological perspective, several marine hosts from different locations around Japan were examined to characterize the parasitism of Anisakis larvae. Chum salmon (Oncorhynchus keta) and Alaska pollock (Gadus chalcogrammus) showed the highest overall prevalence (100%), followed by blue mackerel (Scomber australasicus) (97.5%), Pacific cod (Gadus macrocephalus) (80%), chub mackerel (Scomber japonicus) (60.1%), Japanese flying squid (Todarodes pacificus) (17%) and Japanese pilchard (Sardinops sagax melanostictus) (2%). In Pacific krill (Euphausia pacifica), apart from one Hysterothylacium aduncum larva, no Anisakis specimens were detected. Anisakis simplex sensu stricto was molecularly identified (PCR-RFLP) for the first time in Japanese flying squid and Japanese pilchard distributed in the Northwestern Pacific ocean. That was the most frequent parasitic species detected followed by A. pegreffii, mostly in the western areas of Japan, hybrid genotypes between the two sibling species as well as A. typica and A. berlandi. Surprisingly, A. simplex s.s. was the most abundant species in one batch of chub mackerel from the East China Sea and A. pegreffii was the main species found in one batch from the Pacific coast of Aomori, which seems to indicate that the ranges of these two sibling species might be more variable than previously thought.

Validation of a Commercial Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Anisakis spp. DNA in Processed Fish Products.

Parasites belonging to the Anisakis genera are organisms of interest for human health because they are responsible for the Anisakiasis zoonosis, caused by the ingestion of raw or undercooked fish. Furthermore, several authors have reported this parasite to be a relevant inducer of acute or chronic allergic diseases. In this work, a rapid commercial system based on Loop-Mediated Isothermal Amplification (LAMP) was optimised and validated for the sensitive and rapid detection of Anisakis spp. DNA in processed fish products. The specificity and sensitivity of the LAMP assay for processed fish samples experimentally infected with Anisakis spp. larvae and DNA were determined. The LAMP system proposed in this study was able to give positive amplification for all the processed fish samples artificially contaminated with Anisakis spp., giving sensitivity values equal to 100%. Specificity tests provided no amplification for the Contracaecum, Pseudoterranova, or Hysterothylacium genera and uninfected samples. The limit of detection (LOD) of the LAMP assay proposed was 102 times lower than the real-time PCR method compared. To the best of our knowledge, this is the first report regarding the application of the LAMP assay for the detection of Anisakis spp. in processed fish products. The results obtained indicate that the LAMP assay validated in this work could be a reliable, easy-to-use, and convenient tool for the rapid detection of Anisakis DNA in fish product inspection.

Seasonal trend of Anisakidae infestation in South Mediterranean bluefish.

A total of 1104 fish samples from markets of Sicily were analysed for the detection and species identification of Anisakidae nematodes. The preliminary analysis of the fish samples showed the presence of 2459 larvae. All the fish species revealed different prevalence of infestation, with a maximum of 100% for Lepidopus caudatus and a minimum of 4.5% in Sardina pilchardus. The 80% of the larvae examined by PCR-RFLP analysis belonged to Anisakis pegreffii species. The seasonal infestation trend of Anisakis was evaluated in all the fish sample examined. The results of the seasonal infestation trend showed a marked connection with the ecological aspects of the fish species examined. As far as we know, this work report for the first time important ecological aspects of Lepidopus caudatus specimens of South Mediterranean. This work could be useful to plan a seasonal fishing strategy aimed at reducing the health risks related to Anisakis.

Presence of Anisakidae in commercial fish species imported into the Belgian food markets: A systematic review and meta-analyses.

Anisakidae are marine zoonotic nematodes with most commercial fish species as intermediate hosts. Both public health risks and socio-economic problems are attributed to these larvae. Despite these concerns, the occurrence of Anisakidae in commercial fish species in Belgium remains unknown. Therefore, the main objective of this systematic review was to look into studies assessing the prevalence and intensity (level of infection) of Anisakidae in countries importing fish to the Belgian market. The databases of PubMed, Web of Science, Cordis, Google Scholar, Google, African Journals online and Asia Journals online were searched. Main eligibility criteria were: fish species consumed in Belgium; studies conducted in one of the main importing countries; and the availability of prevalence data. From the original 519 identified studies, 83 were included with data from Spain, Germany, Chile, Denmark, Turkey, France, China, England, Belgium, Norway, Iceland, Senegal and Sweden. Overall results show a widespread occurrence of Anisakidae with a high variability in prevalence between fish species and fishing sea. Cod (Gadus morhua) and Atlantic salmon (Salmo salar), the most consumed fish species in Belgium, have a mean prevalence of 33% and 5% respectively. Of all investigated fishing zones, fish caught in the Northeast Atlantic has the highest rate of infection (68%). Furthermore, higher prevalences were found when looking at the viscera (mean prevalence 59%) compared to the muscle (29%) and with superior techniques such as enzymatic digestion or UV press (46%) compared to candling, the routine method (23%). Farmed fish were found to be the least infected (2%) but were still not Anisakidae free. The widespread presence of Anisakidae and the associated food safety implications indicate the need to further investigate the presence of Anisakidae in fish in the Belgian market.

Presence of Anisakis pegreffii in farmed sea bass (Dicentrarchus labrax L.) commercialized in Southern Italy: A first report.

We examined 151 European sea bass (Dicentrarchus labrax L.) samples from farms and fish markets of Sicily (Southern Italy) for Anisakidae larvae detection. All the samples were examined by visual inspection and modified chloro-peptic digestion. Two nematode larvae were found in the viscera of only one European sea bass sample from a farm located in Greece (FAO 37.3), giving a total prevalence of infestation of 0.7%. No other parasites were found after chloro-peptic digestion of the samples. The larvae were morphologically ascribed, at genus level, to morphotypes I and molecularly identified as Anisakis pegreffii. To the best if our knowledge, this is the first report on the presence of anisakid parasites in farmed European sea bass of Mediterranean Sea. Our findings suggest that the risk of exposure to Anisakidae nematodes in farmed European sea bass remains very low. However, further data on Mediterranean farms are needed to have a detailed risk analysis.

Integrative taxonomy of anisakid nematodes in stranded cetaceans from Brazilian waters: an update on parasite's hosts and geographical records.

Anisakidae are represented mainly by species of Anisakis Dujardin, 1845, Pseudoterranova Krabbe, 1878, and Contracaecum Railliet and Henry, 1913. Integrative taxonomy, based on morphological and genetic approaches, can provide a precise identification of species, increasing the knowledge of their systematics, biology, and evolution. In this study, integrative taxonomy was applied in order to identify, at generic and/or specific taxonomic level, 245 anisakids from 63 cetaceans of 12 different species, which stranded in the Brazilian coast. Parasites from nine hosts were preserved by freezing and, from 54 hosts, were fixed in 70% ethanol (n = 43) or alcohol-formalin-acetic (n = 11) for a period varying from 6 to 19 years. The morphological analysis allowed the identification of 171 specimens at genus level: 120 as Anisakis sp. clade I, 47 as Anisakis sp. clade II, and four as Pseudoterranova sp. From those 15, specimens were identified at species level, seven as Anisakis typica, seven as Anisakis paggiae and one as Anisakis brevispiculata. Moreover, 74 specimens were identified as belonging to Anisakidae family. The genetic analysis based on cox2 gene allowed the identification/confirmation of Anisakis typica (n = 33), Anisakis paggiae (n = 1), and for the first time in the Brazilian coast, Anisakis brevispiculata (n = 4) and Anisakis ziphidarum (n = 1). Additionally, a probably new Anisakis species, genetically close to A. paggiae, is reported infecting a dwarf sperm whale. The results clearly indicated that the preservation of nematodes influenced significantly on the level of taxonomic definition, in both morphological and, mainly, genetic analyses. Pseudoterranova sp. was identified for the first time in the spinner dolphin, Stenella longirostris. The detection of nine A. paggiae specimens, in three Kogia breviceps animals, substantiates the austral occurrence of this species. The identification of A. brevispiculata in Kogia sima and K. breviceps confirms Kogiidae whales as their most common hosts. Anisakis typica was corroborated as a generalist species, the most widely distributed in the Brazilian coast, infecting a broad variety of Delphinidae and Kogiidae, with two new geographical records in Stenella frontalis and Lagenodelphis hosei. The study showed a new scenery of parasite diversity of marine anisakids, updating the Anisakis species distribution.

Gastric lesions in dolphins stranded along the Eastern Adriatic coast.

Stranded cetaceans are often found with gastric lesions associated with the presence of parasites; most frequently, nematodes of the genus Anisakis and the heterophyd digenean trematode Pholeter gastrophilus. In this study, we present histopathology mainly (but not exclusively) related to these 2 parasite species. Macroscopically, lesions associated with the presence of Anisakis spp. were characterised by the presence of ulcers within the gastric mucosa, while the digenean P. gastrophilus was found within large submucosal fibrotic nodules in the gastric wall. Anisakis-induced alterations included severe ulcerative gastritis with mixed inflammatory infiltrate often associated with colonies of bacteria, and mild to moderate granulomatous gastritis with eosinophilic infiltrate. P. gastrophilus-associated lesions were characterised by fibrogranulomatous gastritis with mixed inflammatory infiltrate. Additionally, immunohistochemical (IHC) analysis of P. gastrophilus lesions was consistent with the histopathologic findings, revealing inflammation-mediated stimulation. IHC-positive localisation of CD3+, iNOS+ and caspase-3+ cells suggests intensive accumulation of cytotoxic T-cells, proinflammatory cytokines and execution-phase of cell apoptosis at the parasitized area. In contrast, mechanical damage, rather than visible inflammatory response could be observed at the site of attachment of Braunina cordiformis recorded in 4 animals. Lesions not associated with the presence of parasites were mostly characterised by focal loss of superficial epithelial cells and accumulation of brown hemosiderin-like pigment or fibrous gastritis with lymphoplasmacytic infiltrate. In light of these results, we argue that observed 'tolerant' host-parasite interactions that led toward gastric lesions do not represent the cause of death and stranding of cetaceans included in this study.

Pathogenic Potential of Fresh, Frozen, and Thermally Treated Anisakis spp. Type II (L3) (Nematoda: Anisakidae) after Oral Inoculation into Wistar Rats: A Histopathological Study.

The third-stage (L3) larvae of Anisakis are the etiological agents of human anisakiasis caused by consumption of raw or undercooked seafood infected with anisakid nematodes. Infection with these worms is associated with abdominal pain, nausea, and diarrhea and can lead to massive infiltration of eosinophils and the formation of granulomas in the gastrointestinal tract if the larvae are not removed. Food allergy affects populations worldwide, and despite several reports on the presence of the potentially zoonotic nematodes among edible fishes in Egypt, there are few immunological and molecular studies investigating the epidemiology of these parasites. Anisakidosis, a human infection with nematodes of the family Anisakidae, is caused most commonly by Anisakis spp. In the present study, seventy specimens of the European seabass Dicentrarchus labrax commercialized in Alexandria city along the Mediterranean Sea were acquired during the period from July to December, 2015. Fish were necropsied and dissected to investigate the presence of nematode larvae. Thirty fish (42.9%) of the total were parasitized by nematode larvae which were morphologically identified as Anisakis spp. Type II (L3) according to light and scanning electron microscopy. The pathogenic potential of oral inoculation of fresh, frozen, and thermally treated larvae into Wistar rats was elucidated by histological examination of their thymus and spleen. Results obtained indicated that neither cooling nor freezing of the parasite could destroy their allergenic capacity. So, it is important to create a wider awareness of this potential risk to human health. It is becoming increasingly likely that the impact of Anisakis spp. on human health has been underestimated, and it is perhaps time to consider more sweeping measures than those currently enforced to protect the public health.

Principal component analysis of factors for sensitization to Anisakis spp. in postpartum women.

INTRODUCTION: Immunoreactivity to Anisakis spp. is believed to be associated with frequency of fish intake. The objective of this study was to evaluate, using principal component analysis, the main factors potentially involved in reactivity to these nematodes in postpartum women. METHODS: Retrospective study conducted on a database of 309 postpartum women. All completed a structured questionnaire and had blood samples collected for ELISA analysis of specific immunoglobulins against total Anisakis spp. antigens and assessment of reactivity. Parametric and nonparametric tests were used to assess factors for sensitization in the reactive and nonreactive groups, and a principal component analysis was performed. A Pearson correlation matrix with varimax rotation was used to assess the variables of interest (place of residence, age, number of prenatal visits, type of birth facility, fish intake and frequency, raw fish intake, fish handling, history of allergies). RESULTS: After exclusions, samples from 203 women were assessed. Of these, 52 (25.6%) were reactive for anti-Anisakis IgG. Most women claimed not to handle fish (n = 121) and eat fish only sporadically (n = 71). Significant differences in age were seen between the reactive and nonreactive groups (p = 0.001). The first two components explained 32.55% and 38.94% of variances in the nonreactive and reactive groups respectively. The adjusted matrix assigned greater probabilistic weight to weekly intake frequency (0.804), followed by raw fish intake (0.759), with differences in relation to the nonreactive group. CONCLUSION: Correlation matrices revealed a direct relationship between seroreactivity to Anisakis spp. and frequency of fish intake in a sample of postpartum women.

Organochlorine pesticides and parasites in Mugil incilis collected in Cartagena Bay, Colombia.

Nematode parasites of the Anisakides family are often found in people living in countries where fish is consumed raw or partially cooked. This research shows the histological changes in the liver and spleen of Mugil incilis, collected in Cartagena Bay. These changes are associated with pollution by organochlorine pesticides and their possible influence on the parasite. Organochlorine compounds were extracted using the headspace-solid-phase microextraction (HS-SPME) technique. Residual amounts in the muscle of M. incilis such as ß-HCH, γ-HCH, heptachlor, aldrin, endosulfan, 4,4'-DDE, and dieldrin, among others, were identified by gas chromatography connected to an electron capture detector, indicating that the fauna of Cartagena Bay are exposed to these pollutants. Histological analysis was carried out on liver and spleen samples of M. incilis which were fixed, processed, and embedded in paraffin. The presence of melano-macrophages, granulomes, and trematodes in the liver was the most important changes observed. Larval prevalence for the Anisakis spp. was determined to be 1.6%; for Pseudoterranova spp., 25.3%, and for Contracaecum spp., 57.8%. Other parasites such as acanthocephalans were also reported for a total of 15.3%. Nevertheless, no significant correlation between parasites and organochlorines was found. This study is the first to correlate the presence of organochlorine compounds and histological damage in the liver and spleen of M. incilis, with the presence of parasites in fish from Cartagena Bay (Colombia).

Three Anisakis spp. isolated from toothed whales stranded along the eastern Adriatic Sea coast.

Knowledge concerning cetacean ecology in the Mediterranean is limited but important for sustainable planning and enforcement of appropriate conservation measures. Any information that might help to elucidate their ecology is essential. We explored the population and genetic structures of Anisakis spp. nematodes isolated from four toothed whale species - bottlenose dolphins (Tursiops truncatus), striped dolphins (Stenella coeruleoalba), Risso's dolphins (Grampus griseus) and Cuvier's beaked whales (Ziphius cavirostris) - stranded along the eastern Adriatic Sea coast (1990-2012) to reveal more information on host ecological patterns. Lower parasite prevalence was observed in resident dolphin species compared with occasionally occurring species, as well as in young compared with adult dolphins, indicating different feeding habits related to age. No unequivocal relationship between the biological traits of a host (age, body length, body mass and blubber depth) and Anisakis population parameters was observed. Phylogenetic analysis revealed a new geographical record of Anisakis simplex sensu stricto (1.96%) and Anisakis physeteris (1.31%) in the Adriatic Sea in addition to resident Anisakis pegreffii (96.73%). In an assessment of the Adriatic Sea and oceans worldwide, the genetic structure of Anisakis revealed that A. pegreffii populations do not differ among various final host species but do differ with respect to geographical location in contrast to previously accepted Anisakis panmixia.