Variation in the levels of Anisakis spp. infection in the European anchovy Engraulis encrasicolus (L. 1758) from the Bay of Biscay in the period 2000-2023 (ICES Subarea 8)

doi:10.21203/rs.3.rs-3226968/v1

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Variation in the levels of Anisakis spp. infection in the European anchovy Engraulis encrasicolus (L. 1758) from the Bay of Biscay in the period 2000-2023 (ICES Subarea 8)

https://doi.org/10.21203/rs.3.rs-3226968/v1

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published 13 Jan, 2024

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The European anchovy Engraulis encrasicholus is one of the most commercial important species in the Bay of Biscay (ICES Subarea 8). We analysed the temporal mean abundance of Anisakis spp. larvae species in anchovies from ICES subarea 8 in 2000, 2001, 2014-2016, 2019-2023. Prevalence, mean abundance and intensity were examined by length classes. Prevalence in adults was only 1% in 2000 but increased to 90% in 2014. Since 2015 prevalence has decreased; and 17.6 % of individuals were affected in 2023. Mean abundance showed a similar trend, with a peak of 3.79 nematodes/anchovy in 2014, which fell to 0.21 in 2023. The species A. simplex (s.s.) and A. pegreffii were identified by the PCR/SANGER sequencing and PCR/RLFP size ITS fragments techniques in 2019 and 2020. Anisakis simplex sensu stricto was the most abundant species, accounting by these two techniques for 61 to 64 % of total nematodes in 2019 and 52% in 2020.

Anisakis

anchovy

Bay of Biscay

abundance

prevalence

genetic.

Many of the most important commercial fish species caught by European fleets are potential nematode hosts. The presence of parasitic nematodes in commercial fish periodically causes alarm among consumers and an immediate decline in sales in European markets. Anisakid larvae are the most important group of these parasites; they are considered a human health risk and one of the factors affecting product quality in European fish markets (Buchmann and Mehrdana 2016; Caldeira et al. 2021; Debenedetti et al. 2019; Guan et al. 2021; Levsen et al. 2018b; Pascual et al. 2018; Rodríguez et al. 2018a; Rose 2018) The anisakid larvae presence in fish edible tissues causes health problems when fish products are consumed raw or poorly cooked, leading to gastric or intestinal diseases and food allergies (Adroher-Auroux and Benítez-Rodríguez 2020; Audicana et al. 2002; Audicana et al. 1995; Daschner et al. 2021; Santos et al. 2022). The habitat of the European anchovy (Engraulis encrasicholus) includes the North East Atlantic, the Mediterranean and the Black Sea. In the Atlantic it is a target species caught mainly in the Bay of Biscay by purse seiners during the springtime spawning period. It is a high-value species in southern Europe and is mainly marketed fresh, salted or marinated. In the last ten years landings in the Bay of Biscay fluctuated between 14,530 and 30,773 tons, mainly caught by the Spanish and French fleets (ICES 2021).

The presence of anisakid larvae in anchovies has mainly been documented in the Mediterranean Sea (Ciccarelli et al. 2011; Cipriani et al. 2016; Cipriani et al. 2018a; De Liberato et al. 2013a; de Liberato et al. 2013b; Mladineo et al. 2010; Mladineo et al. 2012; Ozuni et al. 2021; Pekmezci et al. 2014; Piras et al. 2014 ; Rello et al. 2009; Roca-Geronès et al. 2020; Serracca et al. 2014). Although the distribution of Anisakis is worldwide and it is able to infect many commercial predator fish, few studies are available regarding changes in levels of infection with anisakid larvae parasitizing anchovies in North East Atlantic waters (Chirapozu et al. 2006; Domingo-Hernández et al. 2023; Marigómez et al. 2006; Mladineo et al. 2010; Mladineo et al. 2012; Rodríguez et al. 2018a).

Studies on the identification of Anisakis species infecting anchovies in Mediterranean waters have been reported since the last decade (Cavallero et al. 2012; Chaligiannis et al. 2012; Cipriani et al. 2018a; Costa et al. 2016; Meloni et al. 2011; Paoletti et al. 2018). However, there are very few genetic studies on the characterization of different Anisakis species in anchovies from North East Atlantic waters (Rodríguez et al. 2018b).

The aims of this study are 1) to assess the temporal changes in the prevalence, mean abundance and intensity of anisakid larvae infections in anchovy caught in the Bay of Biscay (North East Atlantic, Subarea ICES 8; and 2) to identify the relative proportions of Anisakis species that infect anchovies in the area of study.

Fish sampling

A total of 3586 anchovies were obtained during the spring seasons of 2000, 2001, 2014, 2015 and 2016 from commercial purse seiners in the south-eastern Bay of Biscay (ICES Division 8bc east) and from BIOMAN scientific surveys in 2019, 2020, 2021, 2022 and 2023. In September 2019, 2020 and 2021 a specific sampling plan collected 908 juveniles during the JUVENA survey in the same fishing grounds, which due to shorter length and lower age were analysed separately (Table 1). The anchovies were frozen at -20º C and transported to the laboratory of AZTI-BRTA in Sukarrieta, Bizkaia, Spain. Almost all anchovies (96.6%) were smaller than 17.5 cm that according to the age-length key used as reference in the Bay of Biscay they belonged to age 1 and 2.

Parasitological analysis

Once in the laboratory, the samples were thawed and the individuals sexed, measured and weighed before the parasitological examination.

The nematodes from the muscle and visceral cavity were collected by disintegration of the anchovy tissues with pincers and scalpels and the nematodes from the visceral cavity were extracted with pincers and visually counted with the help of stereoscopic microscopy. The nematodes from the viscera were first collected with pincers and after digested overnight at 37º C in a solution of water, pepsin, HCl and NaCl in a thermostatized bath with agitation, according with the method described by McGladdery (1986). The nematodes were separated from the digested viscera by means of a 1 mm wire-mesh sieve. The number of nematodes found after the digestion of the viscera was generally negligible, which confirmed that the method of dissection and visual inspection in small fish species like the anchovy was accurate.

Infection level was determined by length class and sex according to the parasitological terms defined by (Bush et al. 1997)

Genetic identification of larval nematodes

A sample of 81 and 63 nematodes randomly collected from 100 anchovies each year in 2019 and 2020 were used for identification at species level by two different techniques.

PCR/SANGER Sequencing:

Sanger sequencing was performed based on to the protocol described by Pekmezci et al. (2014).

PCR/RLFP and size of the ITS fragments:

The polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) analysis of internal transcribed spacers of ribosomal DNA was conducted based on the size of the ITS digestion fragments with the with the combination of HinfI/HhaI enzymes in agarose gel electrophoresis (Community Reference Laboratory for Parasites 2010). This technique is able to identify seven species of the genus Anisakis; A. simplex (s.s.), A. pegreffii, A. berlandi, A. ziphidarum, A. physeteris, A. typica, Anisakis sp. and the hybrid genotype A. simplex X pegreffii.

Statistical analysis of the epidemiological data

The analysis of prevalence, abundance and Intensity of each pool of samples was carried out using Quantitative Parasitology (QPweb). To determinate whether there were significant differences in the mean abundance between subareas and years, the non-parametric Mann-Whitney (Wilcoxon) test of comparison of medians (STATGRAPHICS Centurion XVI, V 16.2.04 StatPoint Technologies, Inc.) was applied.

Parasitological analysis: analysis of temporal series

Of the anchovies analysed, 1232 were males and 1164 females; the remaining 1430 were juveniles or could not be sexed. The whole prevalence, mean abundance and intensity and Total Length range of anchovies by year is shown in Table 1.

Table 1

Prevalence (% and 95% CI), mean abundance (mA and 95% CI) and mean intensity (mI and 95% CI) of Anisakis spp collected in the study since the year 2000 in adults and juveniles of *E. encrasicolus*. TL (Total Length)
Year/age	n. hosts examined	mean TL (mm)	range TL (mm)	P (%)	mA	mI
2000/adults	106	148	129–190	0.94 (0.0-0.05)	0.01 (-0.01-0.03)	1.00 (n.a)
2001/adults	390	165	121–201	10.8 (0.08–0.14)	0.12 (0.08–0.15)	1.10 (0.98–1.21)
2014/adults	434	149	111–183	90.3 (0.78–0.93)	3.79 (3.47–4.12)	4.20 (3.86–4.54)
2015/adults	375	146	108–182	40.5 (0.35–0.46)	0.85 (0.70-1.0)	2.11 (1.84–2.37)
2016/adults	175	148	123–180	22.3 (0.17–0.29)	0.28 (0.19–0.37)	1.26 (1.10–1.41)
2019
/adults	366	127	72–170	16.9 (0.13–0.21)	0.22 (0.17–0.29)	1.32 (1.16–1.53)
/juveniles	338	68	29–128	3.6 (0.02–0.06)	0.04 (0.02–0.06)	1.00 (n.a)
2020
/adults	318	110	85–141	16.7 (0.13–0.21)	0.19 (0.14–0.24)	1.13 (1.04–1.23)
/juveniles	480	85	35–145	1.3 (0.01–0.03)	0.01 (0.00-0.03)	1.17 (1.00-1.50)
2021
/adults	233	121	97–157	15.5 (0.11–0.21)	0.16 (0.11–0.21)	1.06 (1.0-1.14)
/juveniles	90	61	25–87	0.0	0.0	0.0
2022/adults	281	132	100–164	22.4 (0.18–0.28)	0.28 (0.22–0.35)	1.25 (1.13–1.41)
2023/adults	240	124	102–148	17.6 (0.13–0.23)	0.21 (0.15–0.29)	1.21 (1.05–1.65)

In the year 2000 the prevalence and the mean abundance in adults were the lowest of the series with values only of and 0.01 respectively. The highest prevalence was recorded in 2014, in which this parameter increased to reach 90.3% with mean abundance of 3.79 (Fig. 1). In 2015 the values of both parasitological variables in adults decreased by a factor of 2.2 and 4.4 respectively. In 2019, 2020, 2021 and 2023 the prevalence remained stable between 15.5% and 17.6% while the mean abundance significantly decreased to 0.16 Anisakis/anchovy in 2021, which is similar to the values recorded in 2001 (P > 0.05). In 2022 and 2023 the abundance showed a slight increase to 0.28 and 0.21 Anisakis/anchovy, respectively.

The analysis of the 908 juveniles in the years 2019–2021 showed much lower parasitic burden compared to adults. The highest prevalence was recorded in 2019, with 3.6% of the analysed juveniles being infected, while in 2021, none of the 90 examined anchovies were infected. The abundance of juveniles was also much lower compared to that of adults in the same year.

To facilitate the analysis of infection levels by length, all the anchovies analysed in the 2000–2023 period were grouped per length classes of 1 cm (Fig. 2). The mean abundance showed a clear increase in the length of the individuals, with values of 1.98 Anisakis/anchovy in the largest length class.

Genetic identification of Anisakis spp. larvae

The results obtained by the two genetic methodologies, PCR/Sanger sequencing and PCR/RFLP-size ITS fragments, were similar. Species of A. pegreffii and A. simplex were identified by both techniques. A. simplex was the most abundant species, accounting 69% and 53% of the total nematodes identified in 2019 and 2020. The proportion of A. pegreffii was 31–33% in 2019 and 47% in 2020. The presence of the nematode Hysterothylacium aduncum was also detected between 7–9% of samples in 2019 and 2% in 2020 (Table 2).

Table 2

proportion of *Anisakis* species and *H. aduncum* (percentage of samples in the total analysed) identified in anchovy by PCR/SANGER Sequencing and PCR/RFLP-size ITS fragments in 2019 and 2020.
2019	PCR/SANGER Sequencing	PCR/RFLP-size ITS fragments
A. simplex	64%	61%
A. pegreffii	28%	30%
H. aduncum	7%	9%
2020
A. simplex	52%	52%
A. pegreffii	47%	46%
H. aduncum	2%	2%

This study compiled information about infection levels of Anisakis spp. in anchovies from the Bay of Biscay (ICES subareas 8 a,b,c,d) during the period from 2000 to 2020, with some gaps in the historical series. However, this study was carried out using individuals caught in the same fishing area and season of the year for the main fishing stock of this species, using the same methodology for nematode collection. The result, comparing anchovies of similar length/age, showed that infection levels by anisakid nematodes were very variable over the course of different years. There are few published reports providing information on the prevalence and mean abundance of nematodes in anchovies in Atlantic waters (Table 3).The information collected from 1988 to 2001 showed that the prevalence in this period was low (from 1–13.1%), with mean abundance in the range of 0.01 and 0.12.

Information provided by Chirapozu et al. (2006) regarding anchovies from Subarea 8 and caught in 1999 showed that mean abundance (0.03) and prevalence (3.5%) were higher than the levels found in 2000 but lower than those recorded in 2001 in this study.

Table 3

Published data on anisakid infection values in anchovies in North East Atlantic waters since 1988. P = prevalence, I = mean intensity; A = mean abundance; TL = Total Length
North East Atlantic
ICES area	Year of analysis	P (%)	A	I	TL range (mm)	TL average (mm)	authors
ICES 8	1988–1989	2.1		2			Pereira Bueno 1992
ICES 9a South	1998–1999	13.1	0.2	1.54			Rello et al 2009
ICES 8*	1999	3.5	0.03	1	< 130 > 150	151	Chirapozu 2006
ICES 8	2000	0.94	0.01	1	129–190	148	Present study
ICES 8	2001	10.8	0.12	1.1	121–201	165	Present study
ICES 8	2003	25–50					Marigomez et al 2006
ICES 8c West	2014	87.1	4.7				Rodríguez et al. 2018b
ICES 8	2014	90.3	3.79	4.2	111–183	149	Present study
ICES 8	2015	40.5	0.85	2.11	108–182	146	Present study
ICES 8c West	2015	41.3	1.18	2.75			Rodríguez et al. 2018b
ICES 8b	2016	22.3	0.28	1.26	123–180	148	Present study
ICES 8b	2019	16.9	0.22	1.32	72–170	127	Present study
ICES 8b	2020	16.7	0.19	1.13	85–141	110	Present study
ICES 8b	2021	90.3	5.95	6.61	134–166	147	Domingo-Hernández et al. 2023
ICES 8b	2021	15.5	0.16	1.06	97–157	121	Present study
ICES 8b	2022	22.4	0.28	1.25	100–164	132	Present study
ICES 8b	2023	17.6	0.21	1.21	102–148	124	Present study
*Cantabrian Sea

The increase trend in both parameters was observed since 2001 and confirmed with the peak of prevalence and abundance in 2014 reported by Rodríguez et al. (2018b) and by the present study in the same year. In 2015 a decrease in the parasitological parameters was observed to levels around 50% lower than the previous year. From 2016 to 2020 the prevalence progressively decreased to 16.7% and the mean abundance to values similar to those found in 2001. In the most recent study in the area Domingo-Hernández et al. (2023), reported in anchovies collected in 2021 a high prevalence of 90.3% and also the highest values of abundance and intensity in the series. The other hand, the prevalence and abundance reported in the present study in 2021 to 2023 were significantly lower than the data reported by Domingo-Hernández et al. (2023), perhaps because the average size of anchovies were slightly smaller.

Contrary to the of long-lived species such gadoids, which accumulate a high number of nematodes during their lives, retained for long periods in different parts of the body (Pascual et al. 2018), the anchovies analysed in this study belong on average to 140 mm adults corresponding to 1 to 2-year old fish. Anchovies in this size range could be a good indicator of parasitic levels in the study area, because they only accumulate the nematodes ingested during one year of life; anchovies of this size sampled in different years might therefore reflect the year-on-year change in mean abundance.

The results published by Rodríguez et al. (2018b) with anchovies collected in 2014 and 2105 in the Bay of Biscay showed that the proportion of A. pegreffi was much lower than the recorded in 2019 and 2020 in the present study. In accordance with the results of these last two years the anchovies in the Bay of Biscay were infected with A. simplex as the most abundant species though the proportion of each species was different in the two years of the study, with a significant increase of A. pegreffii in 2020 (Table 5).

Genetic studies to identify anisakid larvae in anchovies from Mediterranean waters showed that the most common species is A. pegreffii, accounting for 64–100% of total nematodes (Chaligiannis et al. 2012; Cipriani et al. 2016; Cipriani et al. 2018a; Meloni et al. 2011; Paoletti et al. 2018; Piras et al. 2014 ; Serracca et al. 2014), and in much less proportion A. simplex and hybrids of A. pegreffii and A. simplex s.s (Costa et al. 2016; Meloni et al. 2011; Mladineo and Poljak 2014). Conversely, results in genetic identification reported in the Atlantic showed that A. simplex was the most abundant species, with a much higher relative proportion than in Mediterranean anchovies (Table 4). This statistics in anchovies of the Bay of Biscay have an exception in the results published by Dessier et al. (2016) that reported an unusual high abundance of the nematode Hysterothylacium aduncum, being the proportion of A, simplex and A. pefreffii in the total occurrence of nematodes only the 5.7% and 1.7% respectively.

Table 4

Published data on the proportion of *Anisakis* species in anchovies from Mediterranean and North East Atlantic waters
Mediterranean	A. simplex	A. pegreffi	A. pegreffii + A. simplex (s.s.)	authors
Mediterranean		92%	8%	Meloni et al 2011
Aegean Sea		100%		Chaligianis et al 2012
NW Mediterranean		100%		Serracca 2014
W Mediterranean		100%		Piras et al 2104
CW Mediterranean*	18%	64%	18%	Costa et al 2016
Central Adriatic		100%		Cipriani 2016
Mediterranean		100%		Cipriani et al 2018
Tyrrhenian Sea		100%		Paoletti et al 2018
North East Atlantic
Bay of Biscay (ICES 8b)**	77%	23%		Dessier et al. 2016
Bay of Biscay (ICES 8c West)	89.5%	10.5%		Rodríguez et al. 2018b
Bay of Biscay (ICES 8b)	68%	32%		Present study 2019
Bay of Biscay (ICES 8b)	53%	47%		Present study 2020
Bay of Biscay (ICES 8b)	90.3%	9.7%		Domingo-Hernández et al. 2023
* Proportion from the nucleotide identification: A. simplex (n = 2 sequences), A. simplex + A. pegreffii (n = 2 sequences), and A. pegreffii (n = 7 sequences)
** Proportion only from the sum of A. simplex + A. pegreffii

The analysis of infection levels in anchovy over a long historical series of years clearly showed great variations in the prevalence and mean abundance of Anisakis in the Bay of Biscay. Fiorenza et al. (2020) concluded that the recovery of the abundance of local mammal populations may increase the number of definitive hosts and therefore support the seasonal changes observed in parasite populations. In that regard, an ecological niche model of Anisakis occurrence in fish in the North East Atlantic (Diez et al. 2022b) showed that the increased incidence of anisakid nematodes in the North East Atlantic could be a consequence of the increase in the-host fish stocks, especially hake; and in the abundance of definitive hosts (short-beaked common dolphin, striped dolphin) and sea warming in the North East Atlantic, given that the optimal temperature for Anisakis spp. is higher than that of the area. Also, with respect to the food chain and predatory behaviour, potential migrations of fish species like anchovy to latitudes with a higher abundance of parasite eggs and prey such copepods and euphausiids, crustaceans that act as intermediate hosts of anisakid larvae (Nagasawa 1989; Nagasawa 1990; Smith 1983a; Smith 1983b.), could help explain the changes observed in the study area.

The actual changes in the parasite demography could be related not only to ecological, but also to an ‘anthropogenic shortcut’ in the life cycle of the parasite (Mattiucci et al. 2018). The removal and discard of the viscera on board is a widely used method in commercial fleets to help conserve the flesh quality until the fish are landed in port, and also to prevent the post-mortem migration of the nematode larvae from the viscera to the muscle (Šimat et al. 2015). According to some authors, discarding infected viscera into the sea may affect the natural abundance of Anisakis spp. in the environment, as the viscera can be ingested by paratenic (fish) and definitive hosts (marine mammals) and therefore the nematodes could enter again in the life cycle (Abollo et al. 2001; Mattiucci et al. 2018; Mclelland et al. 1990). The results of an experiment with farmed seabasses free of Anisakis larvae which were fed with fresh hake liver pieces naturally infested with alive anisakis showed only a very small proportion of the nematodes ingested with the viscera were able to reinfect the host, most of them being rapidly digested and defecated. The conclusion was that the availability of larvae that could re-enter the life cycle and re-infect a fish host due to the discarding of fish viscera on board could be much less important than commonly believed (Diez et al. 2022a).

The results of study monitoring infection levels in anchovies showed that the mean abundance of larval nematodes accumulated in their bodies is a dynamic situation, resulting in significant mean abundance and prevalence changes over the course of the study period. Anchovies measuring 140 mm, corresponding to an age from one to two years old, may be a good indicator of overall annual nematode larvae mean abundance in the study area, because those anchovies will only accumulate nematodes ingested during one year of life. Even though anchovies in the Bay of Biscay are mainly infected with A. simplex and A. pegreffii, the proportion of Anisakis species is also a variable parameter and changes from year to year.

Acknowledgements This paper is contribution nº xxxx from AZTI, Marine Research, Basque Research and Technology Alliance (BRTA). We are very grateful to all the participants in the ANIFEMP project who helped collect and analyse samples.

Author contribution Guzmán Díez, Guillermo Boyra, Maria Santos, Guillem Chust and Marina Santurtún contributed to the conception and design of the study. Guillermo Boyra and Maria Santos collected the samples GB, MS, Arantza Maceira and Carmen Abaroa analysed the samples and Elisabette Bilbao and Iñaki Mendibil performed the genetic analysis. Guzmán Díez, Guillermo Boyra and Maria Santos analysed the data, Guzmán Díez wrote the manuscript and carried out substantial revisions of the manuscript. All the authors approved the submitted version.

Funding This study was funded by the Directorate of Fisheries and Aquaculture of the Basque Government (Department of Economic Development, Infrastructures and Environment – Vice Dept. of Agriculture, Fisheries and Food Policies, Directorate of Fisheries and Aquaculture), project ANIFEMP (grant number 00001-IRB2018-33) and by the European Union project (Urban Klima 2050) from the LIFE programme (grant number LIFE 18 IPC 000001).

Declarations Conflict of interest The authors have no competing interests to declare that are relevant to the content of this article

Ethical Approval No approval of research ethics committees was required to accomplish the goals of this study because parasitological analysis was conducted with commercial fish species obtained from fish markets and scientific surveys

Data Availability The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request

Abollo E, Gestal C, Pascual S (2001) Anisakis infestation in marine fish and cephalopods from Galician waters: an updated perspective. Parasitol Res 87:492–499 doi:DOI 10.007/s004360100389
Adroher-Auroux JF, Benítez-Rodríguez R (2020) Anisakiasis and Anisakis: An underdiagnosed emerging disease and its main etiological agents. Research in Veterinary Science 132:535–545 doi:https://doi.org/10.1016/j.rvsc.2020.08.003
Audicana M, Ansótegui IJ, De Corres LF, M.W. K (2002) Anisakis simplex: dangerous-dead and alive? Trends in Parasitology 18(1):20–25
Audicana M, Fernadez de Corres LF, Muñoz D, Frenández E, Navarro JA, Dolores del Pozo M (1995) Recurrent anaphylaxis caused by Anisakis simplex parasitizing fish. J Allergy Clin Immunol:558–560
Buchmann K, Mehrdana F (2016) Effects of anisakid nematodes Anisakis simplex (s.l.), Pseudoterranova decipiens (s.l.) and Contracaecum osculatum (s.l.) on fish and consumer health. Food and Waterborne Parasitology 4 13–22
Bush AO, Lafferty KD, Lotz JM, Shostakl AW (1997) Parasitology meets ecology on its own terms: Margolis et al revisited. J Parasitol 83(4):575–583
Caldeira AJR, Alves CPP, Santos MJ (2021) Anisakis notification in fish: An assessment of the cases reported in the European Union rapid alert system for food and feed (RASFF) database. Food control 124 107913
Cavallero S, Ligas A, Bruschi F, D’Amelio S (2012) Molecular identification of Anisakis spp. from fishes collected in the Tyrrhenian Sea (NW Mediterranean). Veterinary Parasitology 187 563–566
Chaligiannis I, Lalle M, Pozio E, Sotiraki S (2012) Anisakidae infection in fish of the Aegean Sea. Veterinary Parasitology 184 362–366
Chirapozu A, Astorkiza A, De la Dehesa F, Moreno V (2006) Estudio sobre la prevalencia e intensidad de parasitación por larvas de nematodos en boquerones frescos (Engraulis encrasicholus). Alimentaria Diciembre 05- Enero 06:52–70
Ciccarelli C, Aliventi A, Di Trani V, Semeraro AM (2011) Assessment of the prevalence of anisakidae larvae prevalence in anchovies in the central Adriatic Sea. AIVI online 1:15–19
Cipriani P, et al. (2016) Larval migration of the zoonotic parasite Anisakis pegreffii (Nematoda: Anisakidae) in European anchovy, Engraulis encrasicolus: Implications to seafood safety. Food Control 59 148–157
Cipriani P, et al. (2018a) Anisakis pegreffii (Nematoda: Anisakidae) in European anchovy Engraulis encrasicolus from the Mediterranean Sea: Fishing ground as a predictor of parasite distribution. Fisheries Research 202:59–68
Community Reference Laboratory for Parasites (2010). Identification of Anisakidae Larvae at the species level by PCR/RFLP. Istituto Superiore di Sanità., 12 pp.
Costa A, et al. (2016) Survey on the presence of A. simplex s.s. and A. pegreffii hybrid forms in Central-Western Mediterranean Sea. Parasitology International 65 696–701
Daschner A, Levsen A, Cipriani P, Cuéllar del Hoyo C (2021) Anisakis allergy: unjustified social alarm versus healthy diet. Fish Parasitology 120:769–771
De Liberato C, et al. (2013a) Presence of anisakid larvae in the European anchovy, Engraulis encrasicolus, fished off the Tyrrhenian coast of central Italy. J Food Prot 76 1643–1648.
de Liberato C, et al. (2013b) Presence of Anisakid Larvae in the European Anchovy, Engraulis encrasicolus, Fished Off the Tyrrhenian Coast of Central Italy. Journal of Food Protection 76(9):1643–1648 doi:doi:10.4315/0362-028X.JFP-13-092
Debenedetti AL, et al. (2019) Prevalence and Risk of Anisakid Larvae in Fresh Fish Frequently Consumed in Spain: An Overview. Fishes 4(13):16 pp. doi:doi:10.3390/fishes4010013
Dessier A, Dupuy C, Trancart T, Audras A, Bustamante P, Gérard C (2016) Low diversity of helminth parasites in Sardina pilchardus and Engraulis encrasicolus (Clupeidae) from the Bay of Biscay. Mar Freshw Res 67:1583–1588 doi:https://doi.org/10.1071/MF15147
Diez G, et al. (2022a) Infection Rate in Seabasses Fed with Viscera Parasitised by Anisakid Larvae. Acta Parasitologica doi: https://doi.org/10.1007/s11686-022-00525-5
Diez G, et al. (2022b) Analysis of potential drivers of spatial and temporal changes in anisakid larvae infection levels in European hake, Merluccius merluccius (L.), from the North East Atlantic fishing grounds. Parasitology Research doi: https://doi.org/10.1007/s00436-022-07446-2
Domingo-Hernández AM, Morales-Yuste M, Buzo-Domínguez S, Adroher FJ, Benítez R (2023) Anisakis infection in anchovies (Engraulis encrasicolus) from Iberian waters, southwestern Europe: Post-mortem larval migration. Research in Veterinary Science doi:https://doi.org/10.1016/j.rvsc.2023.02.007
Fiorenza EA, et al. (2020) It’s a wormy world: Meta-analysis reveals several decades of change in the global abundance of the parasitic nematodes Anisakis spp. and Pseudoterranova spp. in marine fishes and invertebrates. Glob Change Biol 00:1–13 doi:DOI: 10.1111/gcb.15048
Guan A, Van Damme I, Devlieghere F, Gabriël S (2021) Effect of temperature, CO2 and O2 on motility and mobility of Anisakidae larvae. Scientific Reports 11:Article number:4279
ICES (2021). Working Group for the Bay of Biscay and the Iberian Waters Ecoregion (WGBIE). ICES Scientific Reports, 3:48. 1101 pp. https://doi.org/10.17895/ices.pub.8212
Levsen A, et al. (2018b) A survey of zoonotic nematodes of commercial key fish species from major European fishing grounds—Introducing the FP7 PARASITE exposure assessment study. Fisheries Research 202 4–21
Marigómez I, Soto M, Cancio I, Orbea A, Garmendia L, Cajaraville MP (2006) Cell and tissue biomarkers in mussel, and histopathology in hake and anchovy from Bay of Biscay after the Prestige oil spill (Monitoring Campaign 2003). Mar Poll Bull 53:287–304
Mattiucci S, Cipriani P, Levsen A, Paoletti M, Nascetti G (2018) Molecular Epidemiology of Anisakis and Anisakiasis: An Ecological and Evolutionary Road Map. Advances in Parasitology 99:93–263 doi:https://doi.org/10.1016/bs.apar.2017.12.001
McGladdery SE (1986) Anisakis simplex (Nematoda: Anisakidae) infection of the musculature and body cavity of Atlantic herring (Clupea harengus harengus). CanJ Fish Aquat Sci 43(7):1312–1317
Mclelland G, Misra RK, Martell DJ (1990) Larval anisakine nematodes in various fish species from Sable Island bank and vicinity. Can J Fish Aquat Sci 222:83–118
Meloni M, et al. (2011) Molecular characterization of anisakis larvae from fish caught off sardinia. J Parasitol 97(5):908–914 pp.
Mladineo I, Petri M, Segvi T, Dobri N (2010) Scarcity of parasite assemblages in the Adriatic-reared European sea bass (Dicentrarchus labrax) and sea bream (Sparus aurata). Veterinary Parasitology 174 131–138
Mladineo I, Poljak V (2014) Ecology and Genetic Structure of Zoonotic Anisakis spp. from Adriatic Commercial Fish Species. Applied and Environmental Microbiology 80(4):1281–1290
Mladineo I, Simat V, Miletic J, Beck R, Poljak V (2012) Molecular identification and population dynamic of Anisakis pegreffii (Nematoda: Anisakidae Dujardin, 1845) isolated from the European anchovy (Engraulis encrasicolus L.) in the Adriatic Sea. International Journal of Food Microbiology 157 224–229
Nagasawa K (1989). The life cycle of Anisakis simplex. Nematode problems in North Atlantic fish. ICES, CM 1989/F: 6, pp: 3–8
Nagasawa K (1990) The life cycle of Anisakis simplex: a Review. In: Kikuchi HIK (ed) Intestinal Anisakiasis in Japan. SpringerVerlag., p pp: 31–40
Ozuni E, et al. (2021) Prevalence of Anisakis Larvae in Different Fish Species in Southern Albania: Five-Year Monitoring (2016–2020). Appl Sci 11:11528
Paoletti M, Mattiucci S, Colantoni A, Levsen A, Gay M, Giuseppe Nascetti G (2018) Species-specific Real Time-PCR primers/probe systems to identify fish parasites of the genera Anisakis, Pseudoterranova and Hysterothylacium (Nematoda: Ascaridoidea). Fisheries Research 202 38–48
Pascual S, Rodríguez H, Pierce GJ, Hastie LC, González AF (2018) The NE Atlantic European hake: A neglected high exposure risk for zoonotic parasites in European fish markets. Fisheries Research 202 69–78
Pekmezci GZ, et al. (2014) Molecular identification of Anisakis species (Nematoda:Anisakidae) from marine fishes collected in Turkish waters. Veterinary Parasitology 201 82–94
Piras MC, et al. (2014) Molecular and epidemiological data on Anisakis spp. (Nematoda: Anisakidae) in commercial fish caught off northern Sardinia(western Mediterranean Sea). Veterinary Parasitology 203 237–240
Rello FJ, Adroher FJ, Benítez R, Valero A (2009) The fishing area as a possible indicator of the infection by anisakids in anchovies (Engraulis encrasicolus) from southwestern Europe. International Journal of Food Microbiology 129 277–281
Roca-Geronès X, Segovia M, Godínez-González C, Fisa R, Montoliu I (2020) Anisakis and Hysterothylacium species in Mediterranean and North-East Atlantic fishes commonly consumed in Spain: Epidemiological, molecular and morphometric discriminant analysis. International Journal of Food Microbiology () 108642 325 108642
Rodríguez H, Abollo E, González AF, Pascual S, Abaunza P (2018a) Scoring the parasite risk in highly-valuable fish species from southern ICES areas. Fisheries Research 202 134–139
Rodríguez H, González AF, Abollo E, Pascual S (2018b) Re-evaluation of anchovies (Engraulis encrasicolus) as an important risk factor for sensitization to zoonotic nematodes in Spain. Fisheries Research 202 49–58
Rose GA (2018) The fish nematode problem in major European fish stocks. Fisheries Research 202 1–3
Santos MJ, et al. (2022) Preliminary Data on the Occurrence of Anisakis spp. in European Hake (Merluccius merluccius) Caught Off the Portuguese Coast and on Reports of Human Anisakiosis in Portugal. Microorganisms 10, 331 doi:
https://doi.org/10.3390/
Serracca L, et al. (2014) Food safety considerations in relation to Anisakis pegreffii in anchovies (Engraulis encrasicolus) and sardines (Sardina pilchardus) fished off the Ligurian Coast (Cinque Terre National Park, NW Mediterranean). International Journal of Food Microbiology 190 79–83
Šimat V, Miletić J, Bogdanović T, Poljak V, Mladineo I (2015) Role of biogenic amines in the post-mortem migration of Anisakis pegreffii (Nematoda: Anisakidae Dujardin, 1845) larvae into fish fillets. Int J Food Microbiol Dec 2(214):179–186 doi:10.1016/j.ijfoodmicro.2015.08.008.
Smith JW (1983a) Anisakis simplex (Rudolphi, 1809, det. Krabbe, 1878) (Nematoda: Ascaroidea): morphology and morphometry of larvae from euphausiids and fish, and a review of the life-history and ecology.. Journal of Helminthology 57:205–224
Smith JW (1983b.) Larval Anisakis simplex (Rudolphi, 1809, det. Krabbe, 1878) and larval Hysterothylacium sp. (Nematoda: Ascaridoidea) in euphausiids (Crustacea: Malacostraca) in the North-East Atlantic and northern North Sea.. Journal of Helminthology 57:167–177

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Variation in the levels of Anisakis spp. infection in the European anchovy Engraulis encrasicolus (L. 1758) from the Bay of Biscay in the period 2000-2023 (ICES Subarea 8)

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Abstract

Figures

Introduction

Material and Methods

Fish sampling

Parasitological analysis

Genetic identification of larval nematodes

PCR/SANGER Sequencing:

PCR/RLFP and size of the ITS fragments:

Statistical analysis of the epidemiological data

Results

Parasitological analysis: analysis of temporal series

Genetic identification of Anisakis spp. larvae

Discussion

Conclusions

Declarations

References

Additional Declarations

Status:

Journal Publication

Version 1