Papers by Francisco Javier Adroher
Pathogens, 2025
A survey was conducted to assess the impact of on-board gutting in the pouting fishery, Trisopter... more A survey was conducted to assess the impact of on-board gutting in the pouting fishery, Trisopterus luscus (L.), from the Bay of Biscay (area FAO 27.VIII) on the parasite burden of macroscopic ascaridoid nematodes, including anisakids (causing anisakidosis) and raphidascaridids (causing consumer rejection) in these fish. The fish were caught in the Bay of Biscay and collected from the fish market in Granada (southern Spain). Fish larger than 25 cm were gutted on board after capture. A detailed examination of the fish revealed the presence of nematode larvae, which were identified morphologically and molecularly (PCR-RFLP: polymerase chain reaction with restriction fragment polymorphism). Results revealed that ungutted fish harbored only third-stage larvae of ascaridoids (Anisakis and Hysterothylacium) while prevalence reached up to 91%. In contrast, gutted fish exhibited a significant reduction in both the prevalence (36%) and mean abundance (MA, 4.44 vs. 0.91) of these larvae. The prevalence of Anisakis spp. larvae was reduced by over 20%, with a more pronounced reduction in abundance of more than 40% (MA, 1.56 vs. 0.91). Hysterothylacium larvae were completely absent (MA 2.88 vs. 0.00). These findings indicate that gutting, while not highly efficient, lowers Anisakis larvae presence, thereby reducing the risk of anisakiasis to consumers. Additionally, the complete removal of Hysterothylacium larvae enhances the fish’s appearance, making it more appealing and increasing its commercial value, as well as reducing the risk of seizure by health authorities. Further research on these on-board evisceration practices is needed to enhance effectiveness and reduce zoonotic nematodes in commercial fishes.
Diseases of Aquatic Organisms, 2004
Parasitology Research, 1996
Horse mackerel (Trachurus trachurus) from the fish market in Granada, South Spain, were surveyed ... more Horse mackerel (Trachurus trachurus) from the fish market in Granada, South Spain, were surveyed for anisakid nematodes. The fish came from fishing ports all over the country. Larval anisakids were found in 39.4% of the fish examined. In all, 26.1% were infected with third-stage larvae (L3s) of Anisakis simplex; 0.3%, with A. physeteris L3s; 31.1%, with Hysterothylacium aduncum L3s; and 1.7%, with fourth-stage larvae (L4s) of Hysterothylacium sp. Horse mackerel from Mediterranean Sea coast ports (South and Southeast Spain) had the lowest levels of infection and those from Cantabrian Sea coast ports (North Spain) had the highest levels. Variations in infection levels with host size (age) and season of capture were surveyed.& b d y :
Parasitology, 2002
This is the first demonstration of the in vitro development of the 3rd-stage larvae (L3) of Hyste... more This is the first demonstration of the in vitro development of the 3rd-stage larvae (L3) of Hysterothylacium aduncum to the adult. This was achieved in a semi-defined medium that is easy to prepare and to reproduce. The L3, collected from the peritoneal cavity of horse mackerel (Trachurus trachurus), were individually inoculated into RPMI-1640 medium +20% heat-inactivated fetal bovine serum (IFBS). It has been demonstrated that the optimum temperature for development is around 13 °C and is stimulated by the presence of 5% CO2 in the growth atmosphere, increasing the percentage moulting to the 4th larval stage (L4) by 1.9-fold (from 44 to 82%) and the average survival of the nematodes by 1.6 times (from 60 to 96 days). When the larvae were grown at different pHs, optimum development occurred at pH 4.0. Under these conditions, all the larvae moulted to the L4 and more than two-thirds transformed to the adult stage – in which 25–30% of the females laid eggs – and reached an average sur...
Journal of Fish Diseases, 2024
In order to know the risk of anisakiasis (or anisakidosis) by consumption of fish of the genus Mu... more In order to know the risk of anisakiasis (or anisakidosis) by consumption of fish of the genus Mullus from the western Mediterranean Sea in our geographical area, where they are appreciated for their quality, an epidemiological survey was carried out to detect zoonotic or potentially zoonotic nematodes in M. barbatus and M. surmuletus. Although the presence of third larval stage (L3) of anisakids (Anisakis and Contracaecum) has been previously described in these fish, the results showed absence of anisakids and presence of L3 and L4 of raphidascaridids of the genus Hysterothylacium which were molecularly identified as H. fabri. Phylogenetic analysis group them into the Mediterraean Sea clade and far from individuals isolated in the Pacific Ocean. Prevalence was somewhat higher, but not significant, in M. barbatus vs M. surmuletus (72.3 vs 60.0 %), but mean intensity (MI) and mean abundance (MA) parameters were approximately twice as high in M. barbatus as in M. surmuletus (MI 5.8 vs 2.8, p=0.001; MA 4.2 vs 1.7, p<0.001). The presence of the parasite seems to affect these two sympatric species differently. In M. barbatus it seems to affect their growth, as it appreciably reduces the value of allometry coefficient in infected fish (2.78 vs 2.18). On the other hand, in M. surmuletus the infection significantly (p<0.04) affects the Fulton's condition factor, an indicator of the health status of the fish. It can be concluded, that the ingestion of these fish by the population poses no risk of anisakiasis, but the consumer should continue to be urged to follow the rules of prevention against this illness.
#Note that throughout the manuscript the authors use the term ‘red mullets’ in the plural to refer collectively to the two species of the genus Mullus surveyed. When used in the singular, they refer only to Mullus barbatus (red mullet). Not to be confused with mullets, family Mugilidae.
Research in Veterinary Science, 2023
Domingo-Hernández, A.M., Morales-Yuste, M., Buzo-Domínguez, S., Adroher, F.J., Benítez, R. (2023)... more Domingo-Hernández, A.M., Morales-Yuste, M., Buzo-Domínguez, S., Adroher, F.J., Benítez, R. (2023) Research in Veterinary Science, 157: 26-34. Free downloaded available. https://doi.org/10.1016/j.rvsc.2023.02.007
The Anisakis larvae presence in fish for human consumption is a health risk that needs to be monitored. The anchovy is a fish that is highly appreciated by consumers and that can harbour Anisakis. It is thus necessary to periodically evaluate the presence of anisakid larvae in them. So, anchovies from Iberian Peninsula coasts were analysed. Fish examination for macroscopic nematodes showed L3s of both Anisakis type I and Hysterothylacium aduncum. The Anisakis prevalence varies with the catching area and the fish size. The muscle prevalence was 7.45% (mean intensity 1.75; range 1-5). Molecular analysis showed 110 A. simplex s.s. (17 in muscle), 22 A. pegreffii (3) and 7 hybrid genotype individuals (1). Considering that most of the Iberian Peninsula coasts are a sympatry area between these two Anisakis species, it has been observed that A. simplex s.s./A. pegreffii ratio increases from south to north in a clockwise direction. Also, 19 larvae were detected on the fish surface from the Bay of Biscay, indicating the ability of these larvae to migrate after the fish death. The A. simplex s.s./A. pegreffii larvae proportion found on the anchovy surface is similar to the found in viscera and lower than in muscle, suggesting that most of the larvae migrating to the surface must have come from the visceral package. This confirms the importance of removing fish viscera immediately after capture, for those fish species where this is possible. As both species cause anisakiasis/anisakidosis, these data show a real risk to human health, especially in dishes highly prized in Mediterranean countries prepared with raw or semi-raw anchovies.
Pathogens, 2022
Morales-Yuste, M.; Sánchez-Yebra, W.; Garrido, M.; Benítez, R.; Adroher, F.J. (2022). Pathogens 1... more Morales-Yuste, M.; Sánchez-Yebra, W.; Garrido, M.; Benítez, R.; Adroher, F.J. (2022). Pathogens 11 (12): ID1432. Free download available: https://doi.org/10.3390/pathogens11121432.
Abstract: Anisakis larvae and to assess the possible risk of anisakiasis in humans through consumption of this fish. Larvae of the genera Anisakis and Hysterothylacium were identified in the analysis of 128 purchased fish specimens. All Anisakis larvae corresponded to type I. Molecular analysis showed the presence of A. pegreffii, A. simplex s.s., and recombinant genotype between the two. The prevalence of Anisakis was 9.4% with a mean intensity of 1.42, while for Hysterothylacium the values were 12.5% and 1.06. The length and weight of the fish, but not Fulton’s condition factor, varied significantly between infected and uninfected fish. The prevalence of Anisakis increased with fish length, with no fish parasitized with Anisakis measuring less than 15.5 cm (2–2.5 years old), which is probably related to the reported dietary change of these fish at around 2 years of age. Fish not parasitized with any of these nematodes showed positive allometric growth, while those parasitized only with Anisakis showed negative allometric growth. When comparing both groups including only fish ≥ 15.5 cm (the smallest size of Anisakis-infected fish), the difference is shown to be statistically significant (p = 0.01), suggesting that Anisakis infection of spotted flounder negatively affects fish growth even when parasite intensity is low, which may have important economic repercussions. Finally, the low prevalence and, above all, intensity of Anisakis in these fish, as well as the habit of consuming this fish fried in oil in our geographical area, means that the risk of acquiring anisakiasis through consumption of this fish is low.
Keywords: Anisakis; Hysterothylacium; spotted flounder; Citharus linguatula; Gulf of Cadiz; Spain; FAO 27.IXa; anisakiasis; fish growth
Pathogens, 2021
Buzo-Domínguez, S.; Morales‐Yuste, M.; Domingo-Hernández, A.M.; Benítez, R.; Adroher, F.J. (2021)... more Buzo-Domínguez, S.; Morales‐Yuste, M.; Domingo-Hernández, A.M.; Benítez, R.; Adroher, F.J. (2021) Pathogens, 10 (10), ID1302. Free downloadable: https://www.mdpi.com/2076-0817/10/10/1302
The presence of third stage larvae (L3) of Anisakis spp. in wedge sole, Dicologlossa cuneata (Moreau, 1881), purchased in fishmarkets in the city of Granada (Andalusia, southern Spain) was assessed. The wedge sole were caught in two FAO zones: area 27.IXa NE Atlantic (SW Spain coast) and area 34.1.11 CE Atlantic (NW Morocco coast). Only Anisakis larvae, type I, were detected in the largest fish (>20 cm) from the CE Atlantic. These were molecularly identified as A. simplex s.s. The prevalence (P) of Anisakis in this area was 12.5% and the mean intensity (MI) was 1. The presence of Hysterothylacium spp. larvae was also detected in the fish from both areas, with the prevalence being approximately double in the CE Atlantic area (12.5 vs. 5.7). A comparison between the Anisakisinfected and non-infected fish from this area showed that the former were significantly longer than the latter (p < 0.01). These results show that Anisakis parasitization of wedge sole sold in the markets of the city of Granada is of low prevalence and intensity (P = 4.5, MI = 1), especially in those from area 27.IXa (P = 0), indicating that the risk of human infection is low, particularly as this fish is traditionally prepared by deep-frying in oil in Andalusia (southern Spain).
Research in Veterinary Sciences, 2020
Adroher-Auroux, F.J., Benítez-Rodríguez, R., 2020. Res. Vet. Sci. 132, 535–545. https://doi.org/1... more Adroher-Auroux, F.J., Benítez-Rodríguez, R., 2020. Res. Vet. Sci. 132, 535–545. https://doi.org/10.1016/j.rvsc.2020.08.003Anisakiasis or anisakiosis is a human parasitic infection caused by the third-stage larvae (L3) of nematodes of the genus Anisakis, although the term is also used in medical literature for the much less frequent (<3% of cases) infection by L3 of other genera of anisakids, particularly Pseudoterranova. These parasites have a marine lifecycle. Humans are infected by the L3 through ingesting of fish and squid, the intermediate/paratenic hosts. The live larvae generally penetrate the wall of the stomach or intestine causing, among other symptoms, intense pain or allergic symptoms. These are emerging, cosmopolite illnesses. Diagnosis and treatment is usually by endoscopy and extraction and identification of the larvae. Allergic forms are usually diagnosed by prick-test and/or allergen-specific IgE detection and treated with a suitable anti-allergy treatment. The patient is also warned against further consumption of marine fish or squid, as these may be infected with Anisakis. The most common method of prevention is thermal treatment of the entire fish or squid prior to consumption (>60 °C, >1 min or − 20 °C, >24 h). Useful measures for the control of anisakiasis would be to establish a national register of cases, to initiate educational campaigns for the general public and consciousness-raising and training campaigns for health professionals. These would be complemented by control measures for the relevant sectors of the economy: fish operators, fish farming, fishermen, fishmongers, fish industry and catering facilities. Possible genetic predisposition for allergy to Anisakis and the possible relationship between anisakiasis and cancer would also require further investigation.
Foodborne Pathogens and Disease, 2019
Torralbo-Ramírez, V., Molina-Fernández, D., Malagón, D., Benítez, R., Adroher, F.J., 2019. Foodb... more Torralbo-Ramírez, V., Molina-Fernández, D., Malagón, D., Benítez, R., Adroher, F.J., 2019. Foodborne Pathog. Dis. 16, 744–751. https://doi.org/10.1089/fpd.2019.2633
Humans can contract anisakiasis by eating fish or squid containing live larvae of the third stage (L3) of the parasitic nematodes of the genus Anisakis, majorly from Anisakis simplex s.s. and Anisakis pegreffii, sibling species of the A. simplex s.l. complex. Most cases diagnosed molecularly are due to A. simplex s.s., although A. pegreffii has also been identified in human cases. Cathepsins are mostly lysosomal multifunctional cysteine proteases and can participate in the pathogenicity of parasites. Cathepsin B and L activities were investigated in the two sibling species of Anisakis mentioned. L3 and L4 of both species were collected during their in vitro development, and cathepsin activity was determined in the range of pH 4.0–8.5, using specific fluorogenic substrates. The activity detected with the substrate Z-FR-AMC (N-α-benzyloxycarbonyl-L-phenylalanyl-L-arginine-7-amido-4-methyl-coumarin) was identified as cathepsin L (optimum pH = 5.0, range 4.0–6.0, p < 0.001). Activity was highest in L3 freshly collected from fish, especially in A. simplex s.s., and decreased during development, which could be related to virulence, invasion of host tissues, and/or intracellular digestion. Cathepsin B-like activity was not identified with either of the substrates used (Z-RR-AMC [N-α-benzyloxycarbonyl-L-arginyl-L-arginine-7-amido-4-methyl-coumarin] and Z-FR-AMC). With Z-RR-AMC, cleaving activity was detected almost exclusively in L4 of A. simplex s.s. (p < 0.05) with optimum pH = 8.0 (range 7.0–8.5). Assays with class-specific protease inhibitors showed that this activity was mainly due to serine proteases [up to 90% inhibition with 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF)], although metalloproteases (up to 40–45% inhibition with 1,10 phenanthroline) and slight cysteine protease activity (<15% inhibition with E64 [L-trans-epoxysuccinyl-leucylamido-(4-guanidino)-butane]; putative cathepsin B-like) were also detected. These results show differential serine protease activity between sibling Anisakis species, regulated by larval development, at least in A. simplex s.s. The higher cathepsin L and serine protease activities detected in this species could be related to its greater pathogenicity, reported in experimental animals, compared to that of A. pegreffii.
Acta Tropica, 2019
Molina-Fernández, D., Benítez, R., Adroher, F.J., Malagón, D., 2019. Acta Trop. 195, 44–50. https... more Molina-Fernández, D., Benítez, R., Adroher, F.J., Malagón, D., 2019. Acta Trop. 195, 44–50. https://doi.org/10.1016/j.actatropica.2019.04.003
Proteolytic activity was studied in two sibling species of Anisakis (Nematoda: Anisakidae), A. simplex s.s. and A. pegreffii, throughout their in vitro development from third larval stage (L3) from the host fish (L3-0h) to fourth larval stage (L4) obtained in culture. Proteases have a significant role in the lifecycle of the parasite and in the pathogen-host relationship. Proteolytic activity peaks were detected at pH 6.0 and 8.5. Protease activity was detected in all the developmental stages of the two species studied at both pH values. These pH values were used for assaying with specific inhibitors which permitted the determination of metalloprotease activity, and, to a lesser extent, that of serine and cysteine protease. Aspartic protease activity was only detected at pH 6.0. At this pH, L4 larvae showed higher proteolytic activity than L3 larvae in both species (p<0.001), the majority of activity being due to metalloproteases and aspartic proteases, which could be related to nutrition, especially the latter, as occurs in invertebrates. At pH 8.5, proteolytic activity was higher in A. simplex s.s. than in A. pegreffii (p<0.01). At this pH, the majority of activity was due to metalloproteases in all developmental phases of both species, although in L3-0h, the activity of these proteases was significantly higher (p<0.03) in A. simplex s.s. than in A. pegreffii. This could be related to the greater invasive capacity of the former. Serine proteases have frequently been implicated in the invasive capacity and pathogenicity of some parasites.This may be related to the significantly higher activity (p≤0.05) of serine protease in all the larval stages studied of A. simplex at pH 6.0. In summary, there are interspecific differences in proteases that have been related to pathogenesis in nematodes. These differences could thus be contributing to the previously reported differences in pathogenicity between these two Anisakis species.
International Journal of Food Microbiology, 2018
Molina-Fernández, D., Rubio-Calvo, D., Adroher, F.J., Benítez, R., 2018. Int. J. Food Microbiol. ... more Molina-Fernández, D., Rubio-Calvo, D., Adroher, F.J., Benítez, R., 2018. Int. J. Food Microbiol. 282, 49–56. https://doi.org/10.1016/j.ijfoodmicro.2018.05.026
The infection of blue whiting Micromesistius poutassou from the western Mediterranean Sea, off the eastern coast of Spain, with larvae of Anisakis spp. was studied. Between April 2016 and April 2017, 140 fish were analyzed. Total epidemiological data showed that the prevalence of Anisakis spp. was 29.3% and the mean intensity 1.8. Of the 74 larvae collected, 61% were type I and the remaining 39%, type II. Of the former, 91.1% were molecularly identified as Anisakis pegreffii (P=19.3%; MI=1.4), 2.2% as Anisakis simplex s.s. (P=0.7%; MI=1.0), while the rest (6.7%) showed a recombinant genotype between the two (P=2.1%; MI=1.0). All the type II larvae analyzed were molecularly identified as Anisakis physeteris (P=10.0%; MI=2.1). Three fish (2.1%) were found to
have larvae in the muscle, while two were found with 1 larva of A. pegreffii and one with two larvae (1 A. simplex s.s. and 1 A. pegreffii). Statistical analysis showed that the prevalence of Anisakis spp. in blue whiting was higher in spring than in autumn (P<0.001), probably due to the greater size (and age) of the fish and related to factors as diet shift, accumulation with age and higher food intake. Analysis of the data suggested that blue whiting were first infected with Anisakis type I (mean age 2.3years) and later with Anisakis type II (mean age 2.7years),
probably due to the diet changing with age, with the incorporation of the paratenic/intermediate host species of these parasites. In any case, the public health authorities must continue to emphasize the need for suitable thermal treatment (freezing or cooking) of the fish prior to consumption.
Parasitology Research, 2018
Molina-Fernández, D., Adroher, F.J., Benítez, R., 2018. Parasitol. Res. 117, 2095–2103. https://d... more Molina-Fernández, D., Adroher, F.J., Benítez, R., 2018. Parasitol. Res. 117, 2095–2103. https://doi.org/10.1007/s00436-018-5896-5 (Open Access)
The development of the fourth larval stage (L4) of Anisakis physeteris was studied using scanning electron microscopy (SEM), comparing it with third larval stage (L3) recently obtained from the host fish, blue whiting (Micromesistius poutassou), from the western Mediterranean Sea (east coast of Spain, zone FAO 37.1.1). After molting to L4, samples of the parasite were examined at different times in order to observe their development. Following collection of the L4, a small portion was taken from the middle of the larva for molecular identification, confirming in all cases that it was A. physeteris. The anterior and posterior sections of the larvae were prepared for morphological study by SEM. The development of a row of denticles on each of the three prominent lips, almost reaching the buccal commisures, was observed in the L4. Pores of unknown function were found in the upper external part of each lip. Clearly developed cephalic papillae, amphids, and deirids were also observed in L4, while, although present in L3, these were beneath the cuticle. Phasmids were detected in L4 but not in L3. The L4 tail finished in a conical lobe with a blunt point, absent in L3. In the oldest L4, some preanal papillae were observed beneath the cuticle in males, while, in females, the vulva could be seen by light microscopy, apparently still covered by the cuticle.
Parasitology Research, 2017
Molina-Fernández, D., Valles-Vega, I., Hernández-Trujillo, S., Adroher, F.J., Benítez, R., 2017. ... more Molina-Fernández, D., Valles-Vega, I., Hernández-Trujillo, S., Adroher, F.J., Benítez, R., 2017. Parasitol. Res. 116, 2733–2740. https://doi.org/10.1007/s00436-017-5583-y
Eggs obtained from the uteri of female nematodes, genetically identified as Contracaecum multipapillatum s.l., found in a brown pelican (Pelecanus occidentalis) from Bahía de La Paz, Gulf of California, Mexico, were used to study the early developmental stages of this anisakid by scanning electron microscopy (SEM). Egg dimensions were approximately 54 x 45 μm measured by SEM. Observation of the eggs revealed an outer surface of fibrous appearance. The newly hatched larvae were ensheathed and highly motile. Observation with SEM showed that the sheaths of the larvae were striated and revealed an excretory pore and a cleft near the anterior end of the sheath, presumably to facilitate the opening of the sheath for the emergence of the larva. The hatched larvae were placed in nutritive culture medium, where they grew within their sheath, some exsheathing completely two weeks later. The surface patterns of the sheath and the cuticle of the exsheathed larvae were clearly different. Although they did not moult during culture, SEM revealed a morphology typical of third-stage larvae of Contracaecum from fish, as previously observed by optical microscopy. Thus, we suggest that newly hatched larvae from eggs of C. multipapillatum are third larval stage but with sheath of the second larval stage, as occurs in other anisakids. See article in: http://rdcu.be/u8Fw . The final publication is available at link.springer.com
Diseases of Aquatic Organisms, 2017
Valles-Vega, I., Molina-Fernández, D., Benítez, R., Hernández-Trujillo, S., Adroher, F.J., 2017. ... more Valles-Vega, I., Molina-Fernández, D., Benítez, R., Hernández-Trujillo, S., Adroher, F.J., 2017. Dis. Aquat. Organ. 125, 167–178. https://doi.org/10.3354/dao03147 (Open Access)
The initial developmental stages of Contracaecum multipapillatum (von Drasche, 1882) Lucker, 1941 sensu lato were studied using eggs obtained from the uterus of female nematodes (genetically identified) found in a brown pelican Pelecanus occidentalis from Bahía de La Paz (Gulf of California, Mexico). Optical microscopy revealed a smooth or slightly rough surface to the eggs. Egg dimensions were approximately 53 x 43 μm, although when the larvae developed inside their size increased to 66 x 55 μm. Hatching and survival of the larvae was greater at 15 ºC than 24 ºC and increased salinity resulted in a slight increase in hatching but seemed to reduce survival at 24 ºC, but not at 15 ºC. The recently hatched larvae measured 261 x 16 μm within their sheath. When placed in culture medium the larvae grew within their sheath and a small percentage (~2%) exsheathed completely (314 x 19 μm). The larvae continued to grow and develop once they had exsheathed, attaining mean dimensions of 333 x 22 μm. Although they did not moult during culture, optical microscopy revealed a morphology typical of third-stage larvae. Finally, the genetic identity found between the larvae of the parasite from mullet and adult females from the brown pelican suggests a life cycle of C. multipapillatum in which the mullet are involved as intermediate/paratenic hosts and the brown pelicans as final hosts in the geographical area of Bahía de La Paz. Open Access Article. Free Download Available with supplementary material in https://doi.org/10.3354/dao03147 .
Ars Pharmaceutica, 2016
Adroher, F.J., 2016. Ars Pharm. 57, 153–165. https://doi.org/10.4321/S2340-98942016000400002
RES... more Adroher, F.J., 2016. Ars Pharm. 57, 153–165. https://doi.org/10.4321/S2340-98942016000400002
RESUMEN/ABSTRACT
Introducción: Dracunculus medinensis o gusano de Guinea es el mayor nematodo parásito del hombre y causa unas úlceras que generan una importante incapacidad durante 2-4 meses, generalmente, que coincide con los periodos de siembra o recogida de las cosechas. Por ello, esta enfermedad, denominada dracunculosis, provoca importantes pérdidas económicas. En sociedades pobres, las más afectadas, estas pérdidas las mantienen en un círculo vicioso de pobreza del que no pueden salir sin ayuda. La dracunculosis forma parte del grupo de enfermedades tropicales olvidadas que, según la OMS, afecta a más de 1000 millones de pobres del mundo.
Objetivo: Dar a conocer a la sociedad científica hispanohablante el estado actual de la enfermedad y de su lucha contra ella.
Material y métodos: Se ha llevado a cabo una búsqueda bibliográfica en bases de datos científicas, especialmente PubMed (Medline) de la Biblioteca Nacional de Medicina (NML) de Estados Unidos, y se han revisado los informes publicados por los principales organismos e instituciones implicados en la lucha contra la dracunculosis, especialmente The Carter Center, la Organización Mundial de la Salud y Centers for Disease Control and Prevention (CDC) de EE.UU.
Resultados: En 1986 se inició una campaña de erradicación basada en las características epidemiológicas de la enfermedad y en la implicación de los países endémicos, lo que permitía plantear ese objetivo, aprobado por la Asamblea Mundial de la Salud en 1981. Este parásito se distribuía por 18 países de África y 3 de Asia, afectando a 5-10 millones de pobres, según la OMS. El esfuerzo solidario de instituciones, fundaciones, empresas y personal, sobre todo voluntario, ha permitido reducir el número de casos en más del 99,99% tras 30 años de campaña. En 2015 se han contabilizado sólo 22 casos limitados a 4 países de África: Malí, Etiopía, Sudán del Sur y Chad. En 2016 se han detectado 25 casos: 16 en Chad, 3 en Etiopía, 6 en Sudán del Sur y ninguno en Malí (datos provisionales,11-enero-2017).
Conclusión: Se trabaja con la esperanza de que en 2020 la enfermedad esté erradicada del planeta, siendo así la primera de un parásito, mostrándose además que el trabajo de educación sanitaria de la población afectada ha resultado vital para el éxito de la campaña de erradicación.
ABSTRACT
Introduction: Dracunculus medinensis or Guinea worm is the largest nematode parasite in man and causes ulcers that generate a major disability generally for 2-4 months, coinciding with the periods of planting or harvesting of the crops. Therefore, this disease, denominated dracunculiasis, causes important economic losses. In poor societies, those most affected, these losses keep them in a vicious circle of poverty from which they can not leave without help. Dracunculiasis is one of a group of Neglected Tropical Diseases that, according to the WHO, affects more than 1 billion poor people in the world.
Objective: Introduce to the Spanish-speaking scientific community about the current status of the dracunculiasis and its fight against it.
Material and methods: A bibliographic search has been carried out in scientific databases, especially PubMed (Medline) of the National Library of Medicine (NML) of the United States, and the reports published by the main organisms and institutions involved in the fight against dracunculiasis have been reviewed, especially those of The Carter Center, the World Health Organization and Centers for Disease Control and Prevention (CDC).
Results: An eradication campaign was initiated in 1986, based on the epidemiological characteristics of the disease and the involvement of endemic countries, which allowed to raise that objective, adopted by the World Health Assembly in 1981. This parasite was occurring in 18 countries of Africa and 3 of Asia, affecting 5-10 million poor people, according to WHO. The solidarity effort of institutions, foundations, companies and people, especially volunteers, has allowed to reduce the number of cases in more than 99.99% after 30 years of campaign. In 2015, only 22 cases limited to 4 African countries were recorded: Mali, Ethiopia, South Sudan and Chad. In 2016, 25 cases have been detected: 16 in Chad, 3 in Ethiopia, 6 in South Sudan and none in Mali (provisional data1).
Conclusion: It works in the hope that in 2020 the disease will be eradicated from the planet, thus being the first of a parasite. This fight also shows that the health education work on the affected population, carried out jointly by volunteers and officials, is vital to the success of this eradication campaign.
International Journal of Food Microbiology , 2015
Molina-Fernández, D., Malagón, D., Gómez-Mateos, M., Benítez, R., Martín-Sánchez, J., Adroher, F.... more Molina-Fernández, D., Malagón, D., Gómez-Mateos, M., Benítez, R., Martín-Sánchez, J., Adroher, F.J., 2015. Int. J. Food Microbiol. 203, 27–34. https://doi.org/10.1016/j.ijfoodmicro.2015.02.024
The sardine (Sardina pilchardus) is a fish commonly consumed and appreciated in many countries, although they are more likely to be eaten fresh in western Mediterranean countries such as Spain, Portugal, France or Italy. A molecular epidemiological survey of sardines from 5 fishing areas of the Spanish Mediterranean (Málaga, southern Spain) and Atlantic coasts (southern: Cádiz and Isla Cristina; northern: A Coruña and Ondarroa) was carried out to determine the presence of Anisakis spp. larvae. The highest prevalence of these larvae was observed in fish from A Coruña (28.3%), followed by Ondarroa (5%) and Cádiz (2.5%). No Anisakis larvae were found in fish from Málaga and Isla Cristina. Three Anisakis genotypes were identified: Anisakis simplex sensu stricto, Anisakis pegreffii and a hybrid genotype between these two species. A. pegreffii was the most prevalent species in A Coruña (71% of larvae). Only three Anisakis larvae (9% collected larvae) were located in the musculature of sardines: two were identified as A. pegreffii while the other was a hybrid genotype. Sardine infection was associated with fishing area and fish length/weight (length and weight were strongly correlated; Pearson's correlation 0.82; p b 0.001). Risk factor multivariate analysis showed that the risk of infection increases 1.6 times for every additional cm in the length of the sardines from the same fishing area. Comparison of fish of equal length showed that in sardines from A Coruña the risk of parasitization is 11.5 times higher than in those from other fishing areas. Although the risk of infection by Anisakis through consumption of sardines is generally low due to the low epidemiological parameter values (prevalence 10%, mean intensity 1.7 (range 1–5) and mean abundance 0.17), as larger fish are more heavily parasitized, there is an increased risk of infection by Anisakis through consumption of large sardines which are raw or have undergone insufficient treatment (undercooked, smoked, marinated, salted, pickled, freezing,…).
Helminthologia, 2011
Iglesias, L., Benítez, R., Adroher, F.J., Valero, A., 2011. Helminthologia 48, 36–40. https://doi... more Iglesias, L., Benítez, R., Adroher, F.J., Valero, A., 2011. Helminthologia 48, 36–40. https://doi.org/10.2478/s11687-011-0007-5
This survey records the occurrence of larvae of Contracaecum multipapillatum sensu lato (Von Drasche, 1882) Lucker, 1941 (Nematoda, Anisakidae) and adults of Floridosentis mugilis (Machado Filho, 1951) Ward, 1953 (Acanthocephala, Neoechinorhynchidae) in mullet Mugil incilis Hancock, 1830 caught in La Boquilla, Cartagena de Indias (Colombian Caribbean coast). The prevalence of F. mugilis was 45.7 % and of C. multipapillatum s.l. 58.6 %, while 28.6 % of the fish were parasitized by both species. The smallest hosts presented the lowest prevalence of F. mugilis but the highest prevalence of C. multipapillatum s.l.. Prevalence, mean abundance and mean intensity of C. multipapillatum s.l. in M. incilis were all highest in viscera. The prevalence in fish muscle tissue was low (2.9 %).
Veterinary Parasitology, 2011
Malagón, D., Benítez, R., Adroher, F.J., Díaz-López, M., 2011. Vet. Parasitol. 183, 95–102. https... more Malagón, D., Benítez, R., Adroher, F.J., Díaz-López, M., 2011. Vet. Parasitol. 183, 95–102. https://doi.org/10.1016/j.vetpar.2011.07.002
Proteases have a significant role in the life cycle of parasites and the pathogen–host relationship, being regarded as important virulence factors. In the parasitic nematode Hys-terothylacium aduncum proteolytic activity was measured during in vitro development from third larval stage (L3) to mature adult, using DQ red casein as a fluorogenic substrate. Proteolytic activity was detected in all the developmental stages studied and at all pH values within the range employed (2.0–7.5). The assay with specific inhibitors permitted the determination of metalloprotease activity, and, to a lesser extent, that of aspartate-and cysteine-protease. Serine-protease activity was the lowest of those studied. In L3 recently collected from the host fish (L3-0 h), the greatest activity was found at an optimum pH of 4.0 and was mainly inhibited by 1,10-phenathroline (metalloprotease inhibitor). This met-alloprotease activity in L3-0 h (infective stage) may be related to the invasion of the host tissues by this larva. In the other developmental stages, the greatest protease activity was found at pH 5.5, although at pH 4.0 a lower activity peak was detected. On the other hand, our data show that the proteolytic activity of the nematode varies according to the presence of pepsin (an aspartic-protease) in the culture medium. Thus, at pH 4.0, activity was greater in the absence of pepsin, with increasing aspartic-protease activity. Together with the detection of aspartic-, cysteine-and metallo-protease (enzymes involved in digestion in invertebrates) in all the developmental stages of the parasite taking place in the digestive tract of the host fish, this allows us to suggest that the pepsin in the culture medium mimics the predigestion conditions in the habitat of the worm within the host and that the activity detected may have, amongst others, a digestive function.
Diseases of Aquatic Organisms, 2010
Malagón, D., Adroher, F.J., Díaz-López, M., Benítez, R., 2010. Dis. Aquat. Organ. 90, 129–134. ht... more Malagón, D., Adroher, F.J., Díaz-López, M., Benítez, R., 2010. Dis. Aquat. Organ. 90, 129–134. https://doi.org/10.3354/dao02234
Proteases play a vital role in both the life cycle of parasites and the parasite–host relationship and are considered important virulence factors. In the present study, the presence of proteases with collagenolytic activity was investigated in the fish nematode Hysterothylacium aduncum during in vitro development. Collagenolytic activity was found in all studied developmental stages of the nematode (third [L3] and fourth [L4] larval stages and adults). In L3, the activity was maximum at pH 6.5 and, in the other stages, at 7.0. Pepsin is known to favour in vitro development of the worm, but, in this study, collagenolytic activity was shown to be significantly greater when no pepsin was added to the culture medium (at pH 6.5, p = 0.011). At pH 7.0, most activity was observed in the immature adult, after the final moult, suggesting that the collagenolytic activity may be involved in remodelling of the cuticle and in sexual maturity. On the other hand, at pH 6.5, activity may be related to tissue migration by L3 within the host. Using specific inhibitors, it was demonstrated that most of the collagenolytic activity detected in all the developmental stages was due to metalloproteases (40 to 100%), although serine proteases were also detected in L4 and adults (10 to 30%).
Uploads
Papers by Francisco Javier Adroher
#Note that throughout the manuscript the authors use the term ‘red mullets’ in the plural to refer collectively to the two species of the genus Mullus surveyed. When used in the singular, they refer only to Mullus barbatus (red mullet). Not to be confused with mullets, family Mugilidae.
The Anisakis larvae presence in fish for human consumption is a health risk that needs to be monitored. The anchovy is a fish that is highly appreciated by consumers and that can harbour Anisakis. It is thus necessary to periodically evaluate the presence of anisakid larvae in them. So, anchovies from Iberian Peninsula coasts were analysed. Fish examination for macroscopic nematodes showed L3s of both Anisakis type I and Hysterothylacium aduncum. The Anisakis prevalence varies with the catching area and the fish size. The muscle prevalence was 7.45% (mean intensity 1.75; range 1-5). Molecular analysis showed 110 A. simplex s.s. (17 in muscle), 22 A. pegreffii (3) and 7 hybrid genotype individuals (1). Considering that most of the Iberian Peninsula coasts are a sympatry area between these two Anisakis species, it has been observed that A. simplex s.s./A. pegreffii ratio increases from south to north in a clockwise direction. Also, 19 larvae were detected on the fish surface from the Bay of Biscay, indicating the ability of these larvae to migrate after the fish death. The A. simplex s.s./A. pegreffii larvae proportion found on the anchovy surface is similar to the found in viscera and lower than in muscle, suggesting that most of the larvae migrating to the surface must have come from the visceral package. This confirms the importance of removing fish viscera immediately after capture, for those fish species where this is possible. As both species cause anisakiasis/anisakidosis, these data show a real risk to human health, especially in dishes highly prized in Mediterranean countries prepared with raw or semi-raw anchovies.
Abstract: Anisakis larvae and to assess the possible risk of anisakiasis in humans through consumption of this fish. Larvae of the genera Anisakis and Hysterothylacium were identified in the analysis of 128 purchased fish specimens. All Anisakis larvae corresponded to type I. Molecular analysis showed the presence of A. pegreffii, A. simplex s.s., and recombinant genotype between the two. The prevalence of Anisakis was 9.4% with a mean intensity of 1.42, while for Hysterothylacium the values were 12.5% and 1.06. The length and weight of the fish, but not Fulton’s condition factor, varied significantly between infected and uninfected fish. The prevalence of Anisakis increased with fish length, with no fish parasitized with Anisakis measuring less than 15.5 cm (2–2.5 years old), which is probably related to the reported dietary change of these fish at around 2 years of age. Fish not parasitized with any of these nematodes showed positive allometric growth, while those parasitized only with Anisakis showed negative allometric growth. When comparing both groups including only fish ≥ 15.5 cm (the smallest size of Anisakis-infected fish), the difference is shown to be statistically significant (p = 0.01), suggesting that Anisakis infection of spotted flounder negatively affects fish growth even when parasite intensity is low, which may have important economic repercussions. Finally, the low prevalence and, above all, intensity of Anisakis in these fish, as well as the habit of consuming this fish fried in oil in our geographical area, means that the risk of acquiring anisakiasis through consumption of this fish is low.
Keywords: Anisakis; Hysterothylacium; spotted flounder; Citharus linguatula; Gulf of Cadiz; Spain; FAO 27.IXa; anisakiasis; fish growth
The presence of third stage larvae (L3) of Anisakis spp. in wedge sole, Dicologlossa cuneata (Moreau, 1881), purchased in fishmarkets in the city of Granada (Andalusia, southern Spain) was assessed. The wedge sole were caught in two FAO zones: area 27.IXa NE Atlantic (SW Spain coast) and area 34.1.11 CE Atlantic (NW Morocco coast). Only Anisakis larvae, type I, were detected in the largest fish (>20 cm) from the CE Atlantic. These were molecularly identified as A. simplex s.s. The prevalence (P) of Anisakis in this area was 12.5% and the mean intensity (MI) was 1. The presence of Hysterothylacium spp. larvae was also detected in the fish from both areas, with the prevalence being approximately double in the CE Atlantic area (12.5 vs. 5.7). A comparison between the Anisakisinfected and non-infected fish from this area showed that the former were significantly longer than the latter (p < 0.01). These results show that Anisakis parasitization of wedge sole sold in the markets of the city of Granada is of low prevalence and intensity (P = 4.5, MI = 1), especially in those from area 27.IXa (P = 0), indicating that the risk of human infection is low, particularly as this fish is traditionally prepared by deep-frying in oil in Andalusia (southern Spain).
Humans can contract anisakiasis by eating fish or squid containing live larvae of the third stage (L3) of the parasitic nematodes of the genus Anisakis, majorly from Anisakis simplex s.s. and Anisakis pegreffii, sibling species of the A. simplex s.l. complex. Most cases diagnosed molecularly are due to A. simplex s.s., although A. pegreffii has also been identified in human cases. Cathepsins are mostly lysosomal multifunctional cysteine proteases and can participate in the pathogenicity of parasites. Cathepsin B and L activities were investigated in the two sibling species of Anisakis mentioned. L3 and L4 of both species were collected during their in vitro development, and cathepsin activity was determined in the range of pH 4.0–8.5, using specific fluorogenic substrates. The activity detected with the substrate Z-FR-AMC (N-α-benzyloxycarbonyl-L-phenylalanyl-L-arginine-7-amido-4-methyl-coumarin) was identified as cathepsin L (optimum pH = 5.0, range 4.0–6.0, p < 0.001). Activity was highest in L3 freshly collected from fish, especially in A. simplex s.s., and decreased during development, which could be related to virulence, invasion of host tissues, and/or intracellular digestion. Cathepsin B-like activity was not identified with either of the substrates used (Z-RR-AMC [N-α-benzyloxycarbonyl-L-arginyl-L-arginine-7-amido-4-methyl-coumarin] and Z-FR-AMC). With Z-RR-AMC, cleaving activity was detected almost exclusively in L4 of A. simplex s.s. (p < 0.05) with optimum pH = 8.0 (range 7.0–8.5). Assays with class-specific protease inhibitors showed that this activity was mainly due to serine proteases [up to 90% inhibition with 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF)], although metalloproteases (up to 40–45% inhibition with 1,10 phenanthroline) and slight cysteine protease activity (<15% inhibition with E64 [L-trans-epoxysuccinyl-leucylamido-(4-guanidino)-butane]; putative cathepsin B-like) were also detected. These results show differential serine protease activity between sibling Anisakis species, regulated by larval development, at least in A. simplex s.s. The higher cathepsin L and serine protease activities detected in this species could be related to its greater pathogenicity, reported in experimental animals, compared to that of A. pegreffii.
Proteolytic activity was studied in two sibling species of Anisakis (Nematoda: Anisakidae), A. simplex s.s. and A. pegreffii, throughout their in vitro development from third larval stage (L3) from the host fish (L3-0h) to fourth larval stage (L4) obtained in culture. Proteases have a significant role in the lifecycle of the parasite and in the pathogen-host relationship. Proteolytic activity peaks were detected at pH 6.0 and 8.5. Protease activity was detected in all the developmental stages of the two species studied at both pH values. These pH values were used for assaying with specific inhibitors which permitted the determination of metalloprotease activity, and, to a lesser extent, that of serine and cysteine protease. Aspartic protease activity was only detected at pH 6.0. At this pH, L4 larvae showed higher proteolytic activity than L3 larvae in both species (p<0.001), the majority of activity being due to metalloproteases and aspartic proteases, which could be related to nutrition, especially the latter, as occurs in invertebrates. At pH 8.5, proteolytic activity was higher in A. simplex s.s. than in A. pegreffii (p<0.01). At this pH, the majority of activity was due to metalloproteases in all developmental phases of both species, although in L3-0h, the activity of these proteases was significantly higher (p<0.03) in A. simplex s.s. than in A. pegreffii. This could be related to the greater invasive capacity of the former. Serine proteases have frequently been implicated in the invasive capacity and pathogenicity of some parasites.This may be related to the significantly higher activity (p≤0.05) of serine protease in all the larval stages studied of A. simplex at pH 6.0. In summary, there are interspecific differences in proteases that have been related to pathogenesis in nematodes. These differences could thus be contributing to the previously reported differences in pathogenicity between these two Anisakis species.
The infection of blue whiting Micromesistius poutassou from the western Mediterranean Sea, off the eastern coast of Spain, with larvae of Anisakis spp. was studied. Between April 2016 and April 2017, 140 fish were analyzed. Total epidemiological data showed that the prevalence of Anisakis spp. was 29.3% and the mean intensity 1.8. Of the 74 larvae collected, 61% were type I and the remaining 39%, type II. Of the former, 91.1% were molecularly identified as Anisakis pegreffii (P=19.3%; MI=1.4), 2.2% as Anisakis simplex s.s. (P=0.7%; MI=1.0), while the rest (6.7%) showed a recombinant genotype between the two (P=2.1%; MI=1.0). All the type II larvae analyzed were molecularly identified as Anisakis physeteris (P=10.0%; MI=2.1). Three fish (2.1%) were found to
have larvae in the muscle, while two were found with 1 larva of A. pegreffii and one with two larvae (1 A. simplex s.s. and 1 A. pegreffii). Statistical analysis showed that the prevalence of Anisakis spp. in blue whiting was higher in spring than in autumn (P<0.001), probably due to the greater size (and age) of the fish and related to factors as diet shift, accumulation with age and higher food intake. Analysis of the data suggested that blue whiting were first infected with Anisakis type I (mean age 2.3years) and later with Anisakis type II (mean age 2.7years),
probably due to the diet changing with age, with the incorporation of the paratenic/intermediate host species of these parasites. In any case, the public health authorities must continue to emphasize the need for suitable thermal treatment (freezing or cooking) of the fish prior to consumption.
The development of the fourth larval stage (L4) of Anisakis physeteris was studied using scanning electron microscopy (SEM), comparing it with third larval stage (L3) recently obtained from the host fish, blue whiting (Micromesistius poutassou), from the western Mediterranean Sea (east coast of Spain, zone FAO 37.1.1). After molting to L4, samples of the parasite were examined at different times in order to observe their development. Following collection of the L4, a small portion was taken from the middle of the larva for molecular identification, confirming in all cases that it was A. physeteris. The anterior and posterior sections of the larvae were prepared for morphological study by SEM. The development of a row of denticles on each of the three prominent lips, almost reaching the buccal commisures, was observed in the L4. Pores of unknown function were found in the upper external part of each lip. Clearly developed cephalic papillae, amphids, and deirids were also observed in L4, while, although present in L3, these were beneath the cuticle. Phasmids were detected in L4 but not in L3. The L4 tail finished in a conical lobe with a blunt point, absent in L3. In the oldest L4, some preanal papillae were observed beneath the cuticle in males, while, in females, the vulva could be seen by light microscopy, apparently still covered by the cuticle.
Eggs obtained from the uteri of female nematodes, genetically identified as Contracaecum multipapillatum s.l., found in a brown pelican (Pelecanus occidentalis) from Bahía de La Paz, Gulf of California, Mexico, were used to study the early developmental stages of this anisakid by scanning electron microscopy (SEM). Egg dimensions were approximately 54 x 45 μm measured by SEM. Observation of the eggs revealed an outer surface of fibrous appearance. The newly hatched larvae were ensheathed and highly motile. Observation with SEM showed that the sheaths of the larvae were striated and revealed an excretory pore and a cleft near the anterior end of the sheath, presumably to facilitate the opening of the sheath for the emergence of the larva. The hatched larvae were placed in nutritive culture medium, where they grew within their sheath, some exsheathing completely two weeks later. The surface patterns of the sheath and the cuticle of the exsheathed larvae were clearly different. Although they did not moult during culture, SEM revealed a morphology typical of third-stage larvae of Contracaecum from fish, as previously observed by optical microscopy. Thus, we suggest that newly hatched larvae from eggs of C. multipapillatum are third larval stage but with sheath of the second larval stage, as occurs in other anisakids. See article in: http://rdcu.be/u8Fw . The final publication is available at link.springer.com
The initial developmental stages of Contracaecum multipapillatum (von Drasche, 1882) Lucker, 1941 sensu lato were studied using eggs obtained from the uterus of female nematodes (genetically identified) found in a brown pelican Pelecanus occidentalis from Bahía de La Paz (Gulf of California, Mexico). Optical microscopy revealed a smooth or slightly rough surface to the eggs. Egg dimensions were approximately 53 x 43 μm, although when the larvae developed inside their size increased to 66 x 55 μm. Hatching and survival of the larvae was greater at 15 ºC than 24 ºC and increased salinity resulted in a slight increase in hatching but seemed to reduce survival at 24 ºC, but not at 15 ºC. The recently hatched larvae measured 261 x 16 μm within their sheath. When placed in culture medium the larvae grew within their sheath and a small percentage (~2%) exsheathed completely (314 x 19 μm). The larvae continued to grow and develop once they had exsheathed, attaining mean dimensions of 333 x 22 μm. Although they did not moult during culture, optical microscopy revealed a morphology typical of third-stage larvae. Finally, the genetic identity found between the larvae of the parasite from mullet and adult females from the brown pelican suggests a life cycle of C. multipapillatum in which the mullet are involved as intermediate/paratenic hosts and the brown pelicans as final hosts in the geographical area of Bahía de La Paz. Open Access Article. Free Download Available with supplementary material in https://doi.org/10.3354/dao03147 .
RESUMEN/ABSTRACT
Introducción: Dracunculus medinensis o gusano de Guinea es el mayor nematodo parásito del hombre y causa unas úlceras que generan una importante incapacidad durante 2-4 meses, generalmente, que coincide con los periodos de siembra o recogida de las cosechas. Por ello, esta enfermedad, denominada dracunculosis, provoca importantes pérdidas económicas. En sociedades pobres, las más afectadas, estas pérdidas las mantienen en un círculo vicioso de pobreza del que no pueden salir sin ayuda. La dracunculosis forma parte del grupo de enfermedades tropicales olvidadas que, según la OMS, afecta a más de 1000 millones de pobres del mundo.
Objetivo: Dar a conocer a la sociedad científica hispanohablante el estado actual de la enfermedad y de su lucha contra ella.
Material y métodos: Se ha llevado a cabo una búsqueda bibliográfica en bases de datos científicas, especialmente PubMed (Medline) de la Biblioteca Nacional de Medicina (NML) de Estados Unidos, y se han revisado los informes publicados por los principales organismos e instituciones implicados en la lucha contra la dracunculosis, especialmente The Carter Center, la Organización Mundial de la Salud y Centers for Disease Control and Prevention (CDC) de EE.UU.
Resultados: En 1986 se inició una campaña de erradicación basada en las características epidemiológicas de la enfermedad y en la implicación de los países endémicos, lo que permitía plantear ese objetivo, aprobado por la Asamblea Mundial de la Salud en 1981. Este parásito se distribuía por 18 países de África y 3 de Asia, afectando a 5-10 millones de pobres, según la OMS. El esfuerzo solidario de instituciones, fundaciones, empresas y personal, sobre todo voluntario, ha permitido reducir el número de casos en más del 99,99% tras 30 años de campaña. En 2015 se han contabilizado sólo 22 casos limitados a 4 países de África: Malí, Etiopía, Sudán del Sur y Chad. En 2016 se han detectado 25 casos: 16 en Chad, 3 en Etiopía, 6 en Sudán del Sur y ninguno en Malí (datos provisionales,11-enero-2017).
Conclusión: Se trabaja con la esperanza de que en 2020 la enfermedad esté erradicada del planeta, siendo así la primera de un parásito, mostrándose además que el trabajo de educación sanitaria de la población afectada ha resultado vital para el éxito de la campaña de erradicación.
ABSTRACT
Introduction: Dracunculus medinensis or Guinea worm is the largest nematode parasite in man and causes ulcers that generate a major disability generally for 2-4 months, coinciding with the periods of planting or harvesting of the crops. Therefore, this disease, denominated dracunculiasis, causes important economic losses. In poor societies, those most affected, these losses keep them in a vicious circle of poverty from which they can not leave without help. Dracunculiasis is one of a group of Neglected Tropical Diseases that, according to the WHO, affects more than 1 billion poor people in the world.
Objective: Introduce to the Spanish-speaking scientific community about the current status of the dracunculiasis and its fight against it.
Material and methods: A bibliographic search has been carried out in scientific databases, especially PubMed (Medline) of the National Library of Medicine (NML) of the United States, and the reports published by the main organisms and institutions involved in the fight against dracunculiasis have been reviewed, especially those of The Carter Center, the World Health Organization and Centers for Disease Control and Prevention (CDC).
Results: An eradication campaign was initiated in 1986, based on the epidemiological characteristics of the disease and the involvement of endemic countries, which allowed to raise that objective, adopted by the World Health Assembly in 1981. This parasite was occurring in 18 countries of Africa and 3 of Asia, affecting 5-10 million poor people, according to WHO. The solidarity effort of institutions, foundations, companies and people, especially volunteers, has allowed to reduce the number of cases in more than 99.99% after 30 years of campaign. In 2015, only 22 cases limited to 4 African countries were recorded: Mali, Ethiopia, South Sudan and Chad. In 2016, 25 cases have been detected: 16 in Chad, 3 in Ethiopia, 6 in South Sudan and none in Mali (provisional data1).
Conclusion: It works in the hope that in 2020 the disease will be eradicated from the planet, thus being the first of a parasite. This fight also shows that the health education work on the affected population, carried out jointly by volunteers and officials, is vital to the success of this eradication campaign.
The sardine (Sardina pilchardus) is a fish commonly consumed and appreciated in many countries, although they are more likely to be eaten fresh in western Mediterranean countries such as Spain, Portugal, France or Italy. A molecular epidemiological survey of sardines from 5 fishing areas of the Spanish Mediterranean (Málaga, southern Spain) and Atlantic coasts (southern: Cádiz and Isla Cristina; northern: A Coruña and Ondarroa) was carried out to determine the presence of Anisakis spp. larvae. The highest prevalence of these larvae was observed in fish from A Coruña (28.3%), followed by Ondarroa (5%) and Cádiz (2.5%). No Anisakis larvae were found in fish from Málaga and Isla Cristina. Three Anisakis genotypes were identified: Anisakis simplex sensu stricto, Anisakis pegreffii and a hybrid genotype between these two species. A. pegreffii was the most prevalent species in A Coruña (71% of larvae). Only three Anisakis larvae (9% collected larvae) were located in the musculature of sardines: two were identified as A. pegreffii while the other was a hybrid genotype. Sardine infection was associated with fishing area and fish length/weight (length and weight were strongly correlated; Pearson's correlation 0.82; p b 0.001). Risk factor multivariate analysis showed that the risk of infection increases 1.6 times for every additional cm in the length of the sardines from the same fishing area. Comparison of fish of equal length showed that in sardines from A Coruña the risk of parasitization is 11.5 times higher than in those from other fishing areas. Although the risk of infection by Anisakis through consumption of sardines is generally low due to the low epidemiological parameter values (prevalence 10%, mean intensity 1.7 (range 1–5) and mean abundance 0.17), as larger fish are more heavily parasitized, there is an increased risk of infection by Anisakis through consumption of large sardines which are raw or have undergone insufficient treatment (undercooked, smoked, marinated, salted, pickled, freezing,…).
This survey records the occurrence of larvae of Contracaecum multipapillatum sensu lato (Von Drasche, 1882) Lucker, 1941 (Nematoda, Anisakidae) and adults of Floridosentis mugilis (Machado Filho, 1951) Ward, 1953 (Acanthocephala, Neoechinorhynchidae) in mullet Mugil incilis Hancock, 1830 caught in La Boquilla, Cartagena de Indias (Colombian Caribbean coast). The prevalence of F. mugilis was 45.7 % and of C. multipapillatum s.l. 58.6 %, while 28.6 % of the fish were parasitized by both species. The smallest hosts presented the lowest prevalence of F. mugilis but the highest prevalence of C. multipapillatum s.l.. Prevalence, mean abundance and mean intensity of C. multipapillatum s.l. in M. incilis were all highest in viscera. The prevalence in fish muscle tissue was low (2.9 %).
Proteases have a significant role in the life cycle of parasites and the pathogen–host relationship, being regarded as important virulence factors. In the parasitic nematode Hys-terothylacium aduncum proteolytic activity was measured during in vitro development from third larval stage (L3) to mature adult, using DQ red casein as a fluorogenic substrate. Proteolytic activity was detected in all the developmental stages studied and at all pH values within the range employed (2.0–7.5). The assay with specific inhibitors permitted the determination of metalloprotease activity, and, to a lesser extent, that of aspartate-and cysteine-protease. Serine-protease activity was the lowest of those studied. In L3 recently collected from the host fish (L3-0 h), the greatest activity was found at an optimum pH of 4.0 and was mainly inhibited by 1,10-phenathroline (metalloprotease inhibitor). This met-alloprotease activity in L3-0 h (infective stage) may be related to the invasion of the host tissues by this larva. In the other developmental stages, the greatest protease activity was found at pH 5.5, although at pH 4.0 a lower activity peak was detected. On the other hand, our data show that the proteolytic activity of the nematode varies according to the presence of pepsin (an aspartic-protease) in the culture medium. Thus, at pH 4.0, activity was greater in the absence of pepsin, with increasing aspartic-protease activity. Together with the detection of aspartic-, cysteine-and metallo-protease (enzymes involved in digestion in invertebrates) in all the developmental stages of the parasite taking place in the digestive tract of the host fish, this allows us to suggest that the pepsin in the culture medium mimics the predigestion conditions in the habitat of the worm within the host and that the activity detected may have, amongst others, a digestive function.
Proteases play a vital role in both the life cycle of parasites and the parasite–host relationship and are considered important virulence factors. In the present study, the presence of proteases with collagenolytic activity was investigated in the fish nematode Hysterothylacium aduncum during in vitro development. Collagenolytic activity was found in all studied developmental stages of the nematode (third [L3] and fourth [L4] larval stages and adults). In L3, the activity was maximum at pH 6.5 and, in the other stages, at 7.0. Pepsin is known to favour in vitro development of the worm, but, in this study, collagenolytic activity was shown to be significantly greater when no pepsin was added to the culture medium (at pH 6.5, p = 0.011). At pH 7.0, most activity was observed in the immature adult, after the final moult, suggesting that the collagenolytic activity may be involved in remodelling of the cuticle and in sexual maturity. On the other hand, at pH 6.5, activity may be related to tissue migration by L3 within the host. Using specific inhibitors, it was demonstrated that most of the collagenolytic activity detected in all the developmental stages was due to metalloproteases (40 to 100%), although serine proteases were also detected in L4 and adults (10 to 30%).
#Note that throughout the manuscript the authors use the term ‘red mullets’ in the plural to refer collectively to the two species of the genus Mullus surveyed. When used in the singular, they refer only to Mullus barbatus (red mullet). Not to be confused with mullets, family Mugilidae.
The Anisakis larvae presence in fish for human consumption is a health risk that needs to be monitored. The anchovy is a fish that is highly appreciated by consumers and that can harbour Anisakis. It is thus necessary to periodically evaluate the presence of anisakid larvae in them. So, anchovies from Iberian Peninsula coasts were analysed. Fish examination for macroscopic nematodes showed L3s of both Anisakis type I and Hysterothylacium aduncum. The Anisakis prevalence varies with the catching area and the fish size. The muscle prevalence was 7.45% (mean intensity 1.75; range 1-5). Molecular analysis showed 110 A. simplex s.s. (17 in muscle), 22 A. pegreffii (3) and 7 hybrid genotype individuals (1). Considering that most of the Iberian Peninsula coasts are a sympatry area between these two Anisakis species, it has been observed that A. simplex s.s./A. pegreffii ratio increases from south to north in a clockwise direction. Also, 19 larvae were detected on the fish surface from the Bay of Biscay, indicating the ability of these larvae to migrate after the fish death. The A. simplex s.s./A. pegreffii larvae proportion found on the anchovy surface is similar to the found in viscera and lower than in muscle, suggesting that most of the larvae migrating to the surface must have come from the visceral package. This confirms the importance of removing fish viscera immediately after capture, for those fish species where this is possible. As both species cause anisakiasis/anisakidosis, these data show a real risk to human health, especially in dishes highly prized in Mediterranean countries prepared with raw or semi-raw anchovies.
Abstract: Anisakis larvae and to assess the possible risk of anisakiasis in humans through consumption of this fish. Larvae of the genera Anisakis and Hysterothylacium were identified in the analysis of 128 purchased fish specimens. All Anisakis larvae corresponded to type I. Molecular analysis showed the presence of A. pegreffii, A. simplex s.s., and recombinant genotype between the two. The prevalence of Anisakis was 9.4% with a mean intensity of 1.42, while for Hysterothylacium the values were 12.5% and 1.06. The length and weight of the fish, but not Fulton’s condition factor, varied significantly between infected and uninfected fish. The prevalence of Anisakis increased with fish length, with no fish parasitized with Anisakis measuring less than 15.5 cm (2–2.5 years old), which is probably related to the reported dietary change of these fish at around 2 years of age. Fish not parasitized with any of these nematodes showed positive allometric growth, while those parasitized only with Anisakis showed negative allometric growth. When comparing both groups including only fish ≥ 15.5 cm (the smallest size of Anisakis-infected fish), the difference is shown to be statistically significant (p = 0.01), suggesting that Anisakis infection of spotted flounder negatively affects fish growth even when parasite intensity is low, which may have important economic repercussions. Finally, the low prevalence and, above all, intensity of Anisakis in these fish, as well as the habit of consuming this fish fried in oil in our geographical area, means that the risk of acquiring anisakiasis through consumption of this fish is low.
Keywords: Anisakis; Hysterothylacium; spotted flounder; Citharus linguatula; Gulf of Cadiz; Spain; FAO 27.IXa; anisakiasis; fish growth
The presence of third stage larvae (L3) of Anisakis spp. in wedge sole, Dicologlossa cuneata (Moreau, 1881), purchased in fishmarkets in the city of Granada (Andalusia, southern Spain) was assessed. The wedge sole were caught in two FAO zones: area 27.IXa NE Atlantic (SW Spain coast) and area 34.1.11 CE Atlantic (NW Morocco coast). Only Anisakis larvae, type I, were detected in the largest fish (>20 cm) from the CE Atlantic. These were molecularly identified as A. simplex s.s. The prevalence (P) of Anisakis in this area was 12.5% and the mean intensity (MI) was 1. The presence of Hysterothylacium spp. larvae was also detected in the fish from both areas, with the prevalence being approximately double in the CE Atlantic area (12.5 vs. 5.7). A comparison between the Anisakisinfected and non-infected fish from this area showed that the former were significantly longer than the latter (p < 0.01). These results show that Anisakis parasitization of wedge sole sold in the markets of the city of Granada is of low prevalence and intensity (P = 4.5, MI = 1), especially in those from area 27.IXa (P = 0), indicating that the risk of human infection is low, particularly as this fish is traditionally prepared by deep-frying in oil in Andalusia (southern Spain).
Humans can contract anisakiasis by eating fish or squid containing live larvae of the third stage (L3) of the parasitic nematodes of the genus Anisakis, majorly from Anisakis simplex s.s. and Anisakis pegreffii, sibling species of the A. simplex s.l. complex. Most cases diagnosed molecularly are due to A. simplex s.s., although A. pegreffii has also been identified in human cases. Cathepsins are mostly lysosomal multifunctional cysteine proteases and can participate in the pathogenicity of parasites. Cathepsin B and L activities were investigated in the two sibling species of Anisakis mentioned. L3 and L4 of both species were collected during their in vitro development, and cathepsin activity was determined in the range of pH 4.0–8.5, using specific fluorogenic substrates. The activity detected with the substrate Z-FR-AMC (N-α-benzyloxycarbonyl-L-phenylalanyl-L-arginine-7-amido-4-methyl-coumarin) was identified as cathepsin L (optimum pH = 5.0, range 4.0–6.0, p < 0.001). Activity was highest in L3 freshly collected from fish, especially in A. simplex s.s., and decreased during development, which could be related to virulence, invasion of host tissues, and/or intracellular digestion. Cathepsin B-like activity was not identified with either of the substrates used (Z-RR-AMC [N-α-benzyloxycarbonyl-L-arginyl-L-arginine-7-amido-4-methyl-coumarin] and Z-FR-AMC). With Z-RR-AMC, cleaving activity was detected almost exclusively in L4 of A. simplex s.s. (p < 0.05) with optimum pH = 8.0 (range 7.0–8.5). Assays with class-specific protease inhibitors showed that this activity was mainly due to serine proteases [up to 90% inhibition with 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF)], although metalloproteases (up to 40–45% inhibition with 1,10 phenanthroline) and slight cysteine protease activity (<15% inhibition with E64 [L-trans-epoxysuccinyl-leucylamido-(4-guanidino)-butane]; putative cathepsin B-like) were also detected. These results show differential serine protease activity between sibling Anisakis species, regulated by larval development, at least in A. simplex s.s. The higher cathepsin L and serine protease activities detected in this species could be related to its greater pathogenicity, reported in experimental animals, compared to that of A. pegreffii.
Proteolytic activity was studied in two sibling species of Anisakis (Nematoda: Anisakidae), A. simplex s.s. and A. pegreffii, throughout their in vitro development from third larval stage (L3) from the host fish (L3-0h) to fourth larval stage (L4) obtained in culture. Proteases have a significant role in the lifecycle of the parasite and in the pathogen-host relationship. Proteolytic activity peaks were detected at pH 6.0 and 8.5. Protease activity was detected in all the developmental stages of the two species studied at both pH values. These pH values were used for assaying with specific inhibitors which permitted the determination of metalloprotease activity, and, to a lesser extent, that of serine and cysteine protease. Aspartic protease activity was only detected at pH 6.0. At this pH, L4 larvae showed higher proteolytic activity than L3 larvae in both species (p<0.001), the majority of activity being due to metalloproteases and aspartic proteases, which could be related to nutrition, especially the latter, as occurs in invertebrates. At pH 8.5, proteolytic activity was higher in A. simplex s.s. than in A. pegreffii (p<0.01). At this pH, the majority of activity was due to metalloproteases in all developmental phases of both species, although in L3-0h, the activity of these proteases was significantly higher (p<0.03) in A. simplex s.s. than in A. pegreffii. This could be related to the greater invasive capacity of the former. Serine proteases have frequently been implicated in the invasive capacity and pathogenicity of some parasites.This may be related to the significantly higher activity (p≤0.05) of serine protease in all the larval stages studied of A. simplex at pH 6.0. In summary, there are interspecific differences in proteases that have been related to pathogenesis in nematodes. These differences could thus be contributing to the previously reported differences in pathogenicity between these two Anisakis species.
The infection of blue whiting Micromesistius poutassou from the western Mediterranean Sea, off the eastern coast of Spain, with larvae of Anisakis spp. was studied. Between April 2016 and April 2017, 140 fish were analyzed. Total epidemiological data showed that the prevalence of Anisakis spp. was 29.3% and the mean intensity 1.8. Of the 74 larvae collected, 61% were type I and the remaining 39%, type II. Of the former, 91.1% were molecularly identified as Anisakis pegreffii (P=19.3%; MI=1.4), 2.2% as Anisakis simplex s.s. (P=0.7%; MI=1.0), while the rest (6.7%) showed a recombinant genotype between the two (P=2.1%; MI=1.0). All the type II larvae analyzed were molecularly identified as Anisakis physeteris (P=10.0%; MI=2.1). Three fish (2.1%) were found to
have larvae in the muscle, while two were found with 1 larva of A. pegreffii and one with two larvae (1 A. simplex s.s. and 1 A. pegreffii). Statistical analysis showed that the prevalence of Anisakis spp. in blue whiting was higher in spring than in autumn (P<0.001), probably due to the greater size (and age) of the fish and related to factors as diet shift, accumulation with age and higher food intake. Analysis of the data suggested that blue whiting were first infected with Anisakis type I (mean age 2.3years) and later with Anisakis type II (mean age 2.7years),
probably due to the diet changing with age, with the incorporation of the paratenic/intermediate host species of these parasites. In any case, the public health authorities must continue to emphasize the need for suitable thermal treatment (freezing or cooking) of the fish prior to consumption.
The development of the fourth larval stage (L4) of Anisakis physeteris was studied using scanning electron microscopy (SEM), comparing it with third larval stage (L3) recently obtained from the host fish, blue whiting (Micromesistius poutassou), from the western Mediterranean Sea (east coast of Spain, zone FAO 37.1.1). After molting to L4, samples of the parasite were examined at different times in order to observe their development. Following collection of the L4, a small portion was taken from the middle of the larva for molecular identification, confirming in all cases that it was A. physeteris. The anterior and posterior sections of the larvae were prepared for morphological study by SEM. The development of a row of denticles on each of the three prominent lips, almost reaching the buccal commisures, was observed in the L4. Pores of unknown function were found in the upper external part of each lip. Clearly developed cephalic papillae, amphids, and deirids were also observed in L4, while, although present in L3, these were beneath the cuticle. Phasmids were detected in L4 but not in L3. The L4 tail finished in a conical lobe with a blunt point, absent in L3. In the oldest L4, some preanal papillae were observed beneath the cuticle in males, while, in females, the vulva could be seen by light microscopy, apparently still covered by the cuticle.
Eggs obtained from the uteri of female nematodes, genetically identified as Contracaecum multipapillatum s.l., found in a brown pelican (Pelecanus occidentalis) from Bahía de La Paz, Gulf of California, Mexico, were used to study the early developmental stages of this anisakid by scanning electron microscopy (SEM). Egg dimensions were approximately 54 x 45 μm measured by SEM. Observation of the eggs revealed an outer surface of fibrous appearance. The newly hatched larvae were ensheathed and highly motile. Observation with SEM showed that the sheaths of the larvae were striated and revealed an excretory pore and a cleft near the anterior end of the sheath, presumably to facilitate the opening of the sheath for the emergence of the larva. The hatched larvae were placed in nutritive culture medium, where they grew within their sheath, some exsheathing completely two weeks later. The surface patterns of the sheath and the cuticle of the exsheathed larvae were clearly different. Although they did not moult during culture, SEM revealed a morphology typical of third-stage larvae of Contracaecum from fish, as previously observed by optical microscopy. Thus, we suggest that newly hatched larvae from eggs of C. multipapillatum are third larval stage but with sheath of the second larval stage, as occurs in other anisakids. See article in: http://rdcu.be/u8Fw . The final publication is available at link.springer.com
The initial developmental stages of Contracaecum multipapillatum (von Drasche, 1882) Lucker, 1941 sensu lato were studied using eggs obtained from the uterus of female nematodes (genetically identified) found in a brown pelican Pelecanus occidentalis from Bahía de La Paz (Gulf of California, Mexico). Optical microscopy revealed a smooth or slightly rough surface to the eggs. Egg dimensions were approximately 53 x 43 μm, although when the larvae developed inside their size increased to 66 x 55 μm. Hatching and survival of the larvae was greater at 15 ºC than 24 ºC and increased salinity resulted in a slight increase in hatching but seemed to reduce survival at 24 ºC, but not at 15 ºC. The recently hatched larvae measured 261 x 16 μm within their sheath. When placed in culture medium the larvae grew within their sheath and a small percentage (~2%) exsheathed completely (314 x 19 μm). The larvae continued to grow and develop once they had exsheathed, attaining mean dimensions of 333 x 22 μm. Although they did not moult during culture, optical microscopy revealed a morphology typical of third-stage larvae. Finally, the genetic identity found between the larvae of the parasite from mullet and adult females from the brown pelican suggests a life cycle of C. multipapillatum in which the mullet are involved as intermediate/paratenic hosts and the brown pelicans as final hosts in the geographical area of Bahía de La Paz. Open Access Article. Free Download Available with supplementary material in https://doi.org/10.3354/dao03147 .
RESUMEN/ABSTRACT
Introducción: Dracunculus medinensis o gusano de Guinea es el mayor nematodo parásito del hombre y causa unas úlceras que generan una importante incapacidad durante 2-4 meses, generalmente, que coincide con los periodos de siembra o recogida de las cosechas. Por ello, esta enfermedad, denominada dracunculosis, provoca importantes pérdidas económicas. En sociedades pobres, las más afectadas, estas pérdidas las mantienen en un círculo vicioso de pobreza del que no pueden salir sin ayuda. La dracunculosis forma parte del grupo de enfermedades tropicales olvidadas que, según la OMS, afecta a más de 1000 millones de pobres del mundo.
Objetivo: Dar a conocer a la sociedad científica hispanohablante el estado actual de la enfermedad y de su lucha contra ella.
Material y métodos: Se ha llevado a cabo una búsqueda bibliográfica en bases de datos científicas, especialmente PubMed (Medline) de la Biblioteca Nacional de Medicina (NML) de Estados Unidos, y se han revisado los informes publicados por los principales organismos e instituciones implicados en la lucha contra la dracunculosis, especialmente The Carter Center, la Organización Mundial de la Salud y Centers for Disease Control and Prevention (CDC) de EE.UU.
Resultados: En 1986 se inició una campaña de erradicación basada en las características epidemiológicas de la enfermedad y en la implicación de los países endémicos, lo que permitía plantear ese objetivo, aprobado por la Asamblea Mundial de la Salud en 1981. Este parásito se distribuía por 18 países de África y 3 de Asia, afectando a 5-10 millones de pobres, según la OMS. El esfuerzo solidario de instituciones, fundaciones, empresas y personal, sobre todo voluntario, ha permitido reducir el número de casos en más del 99,99% tras 30 años de campaña. En 2015 se han contabilizado sólo 22 casos limitados a 4 países de África: Malí, Etiopía, Sudán del Sur y Chad. En 2016 se han detectado 25 casos: 16 en Chad, 3 en Etiopía, 6 en Sudán del Sur y ninguno en Malí (datos provisionales,11-enero-2017).
Conclusión: Se trabaja con la esperanza de que en 2020 la enfermedad esté erradicada del planeta, siendo así la primera de un parásito, mostrándose además que el trabajo de educación sanitaria de la población afectada ha resultado vital para el éxito de la campaña de erradicación.
ABSTRACT
Introduction: Dracunculus medinensis or Guinea worm is the largest nematode parasite in man and causes ulcers that generate a major disability generally for 2-4 months, coinciding with the periods of planting or harvesting of the crops. Therefore, this disease, denominated dracunculiasis, causes important economic losses. In poor societies, those most affected, these losses keep them in a vicious circle of poverty from which they can not leave without help. Dracunculiasis is one of a group of Neglected Tropical Diseases that, according to the WHO, affects more than 1 billion poor people in the world.
Objective: Introduce to the Spanish-speaking scientific community about the current status of the dracunculiasis and its fight against it.
Material and methods: A bibliographic search has been carried out in scientific databases, especially PubMed (Medline) of the National Library of Medicine (NML) of the United States, and the reports published by the main organisms and institutions involved in the fight against dracunculiasis have been reviewed, especially those of The Carter Center, the World Health Organization and Centers for Disease Control and Prevention (CDC).
Results: An eradication campaign was initiated in 1986, based on the epidemiological characteristics of the disease and the involvement of endemic countries, which allowed to raise that objective, adopted by the World Health Assembly in 1981. This parasite was occurring in 18 countries of Africa and 3 of Asia, affecting 5-10 million poor people, according to WHO. The solidarity effort of institutions, foundations, companies and people, especially volunteers, has allowed to reduce the number of cases in more than 99.99% after 30 years of campaign. In 2015, only 22 cases limited to 4 African countries were recorded: Mali, Ethiopia, South Sudan and Chad. In 2016, 25 cases have been detected: 16 in Chad, 3 in Ethiopia, 6 in South Sudan and none in Mali (provisional data1).
Conclusion: It works in the hope that in 2020 the disease will be eradicated from the planet, thus being the first of a parasite. This fight also shows that the health education work on the affected population, carried out jointly by volunteers and officials, is vital to the success of this eradication campaign.
The sardine (Sardina pilchardus) is a fish commonly consumed and appreciated in many countries, although they are more likely to be eaten fresh in western Mediterranean countries such as Spain, Portugal, France or Italy. A molecular epidemiological survey of sardines from 5 fishing areas of the Spanish Mediterranean (Málaga, southern Spain) and Atlantic coasts (southern: Cádiz and Isla Cristina; northern: A Coruña and Ondarroa) was carried out to determine the presence of Anisakis spp. larvae. The highest prevalence of these larvae was observed in fish from A Coruña (28.3%), followed by Ondarroa (5%) and Cádiz (2.5%). No Anisakis larvae were found in fish from Málaga and Isla Cristina. Three Anisakis genotypes were identified: Anisakis simplex sensu stricto, Anisakis pegreffii and a hybrid genotype between these two species. A. pegreffii was the most prevalent species in A Coruña (71% of larvae). Only three Anisakis larvae (9% collected larvae) were located in the musculature of sardines: two were identified as A. pegreffii while the other was a hybrid genotype. Sardine infection was associated with fishing area and fish length/weight (length and weight were strongly correlated; Pearson's correlation 0.82; p b 0.001). Risk factor multivariate analysis showed that the risk of infection increases 1.6 times for every additional cm in the length of the sardines from the same fishing area. Comparison of fish of equal length showed that in sardines from A Coruña the risk of parasitization is 11.5 times higher than in those from other fishing areas. Although the risk of infection by Anisakis through consumption of sardines is generally low due to the low epidemiological parameter values (prevalence 10%, mean intensity 1.7 (range 1–5) and mean abundance 0.17), as larger fish are more heavily parasitized, there is an increased risk of infection by Anisakis through consumption of large sardines which are raw or have undergone insufficient treatment (undercooked, smoked, marinated, salted, pickled, freezing,…).
This survey records the occurrence of larvae of Contracaecum multipapillatum sensu lato (Von Drasche, 1882) Lucker, 1941 (Nematoda, Anisakidae) and adults of Floridosentis mugilis (Machado Filho, 1951) Ward, 1953 (Acanthocephala, Neoechinorhynchidae) in mullet Mugil incilis Hancock, 1830 caught in La Boquilla, Cartagena de Indias (Colombian Caribbean coast). The prevalence of F. mugilis was 45.7 % and of C. multipapillatum s.l. 58.6 %, while 28.6 % of the fish were parasitized by both species. The smallest hosts presented the lowest prevalence of F. mugilis but the highest prevalence of C. multipapillatum s.l.. Prevalence, mean abundance and mean intensity of C. multipapillatum s.l. in M. incilis were all highest in viscera. The prevalence in fish muscle tissue was low (2.9 %).
Proteases have a significant role in the life cycle of parasites and the pathogen–host relationship, being regarded as important virulence factors. In the parasitic nematode Hys-terothylacium aduncum proteolytic activity was measured during in vitro development from third larval stage (L3) to mature adult, using DQ red casein as a fluorogenic substrate. Proteolytic activity was detected in all the developmental stages studied and at all pH values within the range employed (2.0–7.5). The assay with specific inhibitors permitted the determination of metalloprotease activity, and, to a lesser extent, that of aspartate-and cysteine-protease. Serine-protease activity was the lowest of those studied. In L3 recently collected from the host fish (L3-0 h), the greatest activity was found at an optimum pH of 4.0 and was mainly inhibited by 1,10-phenathroline (metalloprotease inhibitor). This met-alloprotease activity in L3-0 h (infective stage) may be related to the invasion of the host tissues by this larva. In the other developmental stages, the greatest protease activity was found at pH 5.5, although at pH 4.0 a lower activity peak was detected. On the other hand, our data show that the proteolytic activity of the nematode varies according to the presence of pepsin (an aspartic-protease) in the culture medium. Thus, at pH 4.0, activity was greater in the absence of pepsin, with increasing aspartic-protease activity. Together with the detection of aspartic-, cysteine-and metallo-protease (enzymes involved in digestion in invertebrates) in all the developmental stages of the parasite taking place in the digestive tract of the host fish, this allows us to suggest that the pepsin in the culture medium mimics the predigestion conditions in the habitat of the worm within the host and that the activity detected may have, amongst others, a digestive function.
Proteases play a vital role in both the life cycle of parasites and the parasite–host relationship and are considered important virulence factors. In the present study, the presence of proteases with collagenolytic activity was investigated in the fish nematode Hysterothylacium aduncum during in vitro development. Collagenolytic activity was found in all studied developmental stages of the nematode (third [L3] and fourth [L4] larval stages and adults). In L3, the activity was maximum at pH 6.5 and, in the other stages, at 7.0. Pepsin is known to favour in vitro development of the worm, but, in this study, collagenolytic activity was shown to be significantly greater when no pepsin was added to the culture medium (at pH 6.5, p = 0.011). At pH 7.0, most activity was observed in the immature adult, after the final moult, suggesting that the collagenolytic activity may be involved in remodelling of the cuticle and in sexual maturity. On the other hand, at pH 6.5, activity may be related to tissue migration by L3 within the host. Using specific inhibitors, it was demonstrated that most of the collagenolytic activity detected in all the developmental stages was due to metalloproteases (40 to 100%), although serine proteases were also detected in L4 and adults (10 to 30%).
The nematodes are, after the insects, the group of organisms with the largest number of species identified. They include members of great medical, veterinary and agricultural significance, making this group one of the most important animal parasites. However there are many gaps in our knowledge of them. For example, there is still not a single nematode species for which we have detailed knowledge of feeding, digestion and nutritional requirements, showing that there are still many aspects to be learned about nutrition in nematodes [1]. Our understanding of the process of protein digestion, a very important function in the biology of any organism, is still poor since our knowledge is composed of fragmentary data for different groups of nematodes. It is believed that peptidases are essential during the development process and in the most critical moments of parasite-host interactions, and are thus directly involved with the growth and survival of the parasite. Their identification and characterization are important for basic understanding of the biology of the parasite, and their relevance to parasitic nematodes as virulence factors is clear. Consequently, peptidases are currently viewed as potential targets for vaccines, drugs and serodiagnosis. Despite this, in most cases, the precise physiological functions of peptidases in parasites are not known [2]. Peptidases comprise a large class of hydrolytic enzymes in parasitic nematodes, participating in nutrition through digestion of host proteins [3]. ...
Summary. An epidemiological study has been undertaken of ascarids in 914 fishes for human consumption for the inhabitants of Granada. Anisakis simplex (67.9%), Anisakis sp. (2.3%) and Hysterothylacium aduncum (22.4%) have been detected in the host Micromesistius poutassou. Anisakis simplex (27.2%) , Anisakis physeteris (0.3%), Hysterothylacium aduncum (36.7%) and Hysterothylacium sp (1.3%) have been detected in the host Trachurus trachurus. Anisakis simplex (0.9%), Hysterothylacium aduncum (3.5%) Hysterothylacium sp (0.3%) have been found in the Sardina pilchardus. A. simplex have been found in the muscle of 29.1% Micromesistius poutassou, only in 1.0% of Trachurus trachurus and only in 0.3 % of Micromesistius poutassou.
Resumen. Se ha llevado a cabo una encuesta epidemiológica sobre los ascáridos de 914 peces destinados al consumo de la población de Granada. En el hospedador Micromesistius poutassou (bacaladilla) se ha encontrado un índice de parasitación de 67,9% para Anisakis simplex, 2,3 % Anisakis sp. y 22,4% para Hysterothylacium aduncum. En el hospedador Trachurus trachurus (jurel) se ha hallado un índice de
parasitación de 27,2% para Anisakis simplex, 0,3% para Anisakis physeteris, el 36,7% para Hysterothylacium aduncum y el 1,3% Hysterothylacium sp. En el hospedador Sardina pilchardus (sardina) el índice de parasitación observado ha sido de 0,9% para Anisakis simplex, 3,5% para Hysterothylacium aduncum y 0,3% para Hysterothylacium sp. La localización de parásitos en la musculatura de los peces es: A. simplex en un 29,1% de las bacaladillas y 1,0% en jureles. Anisakis sp. 0,3% en bacaladillas.
Anisakis simplex is a nematode of the family Anisakidae which parasitizes a large number of fishes, crustaceans and cephalopods, as intermediate/paratenic hosts, and cetaceans, as final hosts. This parasite affects the man causing the human anisakiasis. The man is usually infected by eating raw, marinated or undercooked fish and squid with third larval stage (L3) of A. simplex. Anisakiasis is a concern of public health in those countries where the fish is consumed raw. For example in Japan, 2000-3000 new cases per year are recorded (Umehara et al., 2007). Also in Spain, new cases every year are recorded and three cases caused by sardines ingestion were described in 1992 (Barros et al., 1992; López-Vélez et al., 1992). The presence of Anisakis larvae in commercial fishes is frequent, especially in fishes such as European hake (Merluccius merluccius) with a prevalence of 87.97% (Valero et al. 2006), blue whiting (Micromesistius poutassou) 81.7% (Martín-Sánchez et al., 2005) and horse mackerel (Trachurus trachurus) 43.9% (Sanmartín et al., 1989).
Anisakis simplex is a parasite ascaridoid nematode, main etiological agent of the parasitic illness known as human anisakiasis, which affects the gastrointestinal tract. The natural final hosts are cetaceans. The intermediate/paratenic hosts are crustaceans, fishes, and cephalopods, which harbor the third larval stage (L3) of this nematode. When the
human, who acts as an accidental host, consumes these raw, marinated or undercooked hosts harboring L3 of Anisakis, has a high risk of acquiring human anisakiasis.
Cita:
Valero A.*, Mira-Gutiérrez J., Pérez-Cano R., Sanz-Domínguez J., Geraldía-Lama M., García-Herruzo J., García-Martos P., Adroher F.J.* (1992). Descripción de un caso de anisakiosis humana. IX Reunión Científica de la Asociación de Parasitólogos Españoles, León, 1-2 October 1992, p. 69.
Resumen: La lisa rayada (Mugil cephalus) se consume con frecuencia entre la población mexicana debido a su bajo costo y a la abundancia con la que se encuentra en las costas de este país. En este trabajo se ha realizado un estudio parasitológico sobre este pescado procedente de las costas de Baja California Sur (México) y se han encontrado dos tipos de parásitos: nematodos que correspondieron a L3 de Contracaecum sp., y acantocéfalos que correspondieron a estadios juveniles y adultos de Floridosentis elongatus (=F. mugilis). Los nematodos se encontraron parasitando al 100% de las lisas en localizaciones diversas como hígado, gónadas, mesenterios intestinales, musculatura y riñón, siendo este último el más afectado. Los acantocéfalos se encontraron siempre en el lumen intestinal causando daños al hospedador como oclusiones y perforaciones intestinales. El porcentaje de parasitación varió según la talla y sexo del hospedador en ambos parásitos. En diversos trabajos se ha implicado a las larvas de Contracaecum spp. como causa de anisakidosis humana (1)(2). Esta circunstancia y el hecho de que la población mexicana consume la lisa ahumada y en forma de cebiche (pescado crudo con limón), justifican este estudio. También se ha ensayado la supervivencia de las L3 de Contracaecum sp. en diversas condiciones de mantenimiento, pudiendo destacar la supervivencia de hasta 3 días a 15ºC en zumo de limón.