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The European conger, Conger conger, is a benthic marine fish species with a geographical distribution extending through the northeastern Atlantic and the Mediterranean. Despite being extensively distributed and widely appreciated by Spanish consumers, studies regarding parasite presence in this fish are scarce. In the present work, a hundred and eight specimens from the Mediterranean coast of northeastern Spain (Catalan waters) were surveyed for the presence of nematode parasites. Several species were morphologically identified: third-stage larvae of Anisakis type I (sensu Berland, 1961) (n = 131), third-stage larvae of Anisakis type II (sensu Berland, 1961) (n = 18), third- and fourth-stage larvae and adults of Hysterothylacium spp. (n = 48), adults of Cucullanus sp. (n = 391), and adults of Cristitectus congeri (n = 69). Moreover, some fish and decapode species were also observed as part of the host's diet, with the most detected preys being Micromesistius poutassou, Sardina pilchardus, Macropipus sp., and Goneplax rhomboides. This represents the first survey of nematode parasites infecting C. conger from the northeastern Spanish Mediterranean waters. Among the parasite species detected, the presence of Anisakis species should be highlighted as the ingestion of C. conger parasitized with these larvae could potentially lead to anisakiasis in consumers. [ABSTRACT FROM AUTHOR]

The epidemiology behind multi-host/multi-parasite systems is particularly interesting to investigate for a better understanding of the complex dynamics naturally occurring in wildlife populations. We aimed to approach the naturally occurring polyparasitism of gastrointestinal nematodes in a sympatric wild ruminant scenario present in south-east Spain. To this end, the gastrointestinal tract of 252 wild ruminants of four different species (red deer, Cervus elaphus; mouflon, Ovis aries musimon; Iberian ibex, Capra pyrenaica and fallow deer, Dama dama) were studied in Cazorla, Segura y Las Villas Natural Park (Andalusia, Spain). Of the analysed animals, 81.52% were positive for parasite infection and a total of 29 nematode species were identified. Out of these, 25 species were detected in at least two host species and 11 parasitized all ruminant species surveyed. The multi-host interaction between these nematodes and the four host species is discussed under the perspective of host family-based differences. [ABSTRACT FROM AUTHOR]

Background: Dirofilaria immitis is responsible for heartworm disease in dogs in endemic areas worldwide. Screening for this infection is done by blood tests. Antigen testing is the most sensitive method to detect an infection with adult (female) worms. Microscopic examination of a blood smear or Knott's test can be used to detect circulating microfilariae, the infective larvae. To increase the sensitivity of the antigen test by decreasing the false negative test results, heating of the blood sample has been recommended in recent guidelines. Heating is believed to remove blocking immune-complexes. Circulating microfilariae are not specific findings for heartworm infection, as other nematodes (among others, Acanthocheilonema dracunculoides) can also result in microfilaremia. Although the type of microfilariae cannot be determined by microscopy alone, real-time PCR can reliably identify the infecting nematode species. Correct identification of the parasite is of major importance, as an infection with D. immitis requires antiparasitic therapy, whereas A. dracunculoides is thought to be a clinically irrelevant coincidental finding. The present case report describes a microfilaremic dog where the initial antigen test for D. immitis turned positive after heat treatment, whereas real-time PCR revealed that the microfilariae were A. dracunculoides (syn. Dipetalonema dracunculoides). Results: A circa 5-year old, asymptomatic Spanish mastiff dog was referred for heartworm therapy because microfilariae were found via a screening blood test. The dog was recently imported to the Netherlands from Spain, where it had been a stray dog. Antigen tests on a plasma sample for D. immitis were performed with three different test kits, which all turned out to be negative. However, heat treatment of two of these samples were carried out and both of them led to a positive antigen test result. Real-time PCR showed that the circulating microfilariae belonged to A. dracunculoides species. Three administrations of moxidectin spot-on at monthly intervals resulted in a negative antigen and a negative Knott's tests one month after the last treatment. Conclusions: We conclude that heat treatment of initially negative blood samples for D. immitis could lead to false positive antigen test results if the dog is infected with A. dracunculoides. [ABSTRACT FROM AUTHOR]

Spirocerca vulpis is a parasitic nematode of red foxes associated with gastric nodule formation. Since its description in 2018, this species has been reported in red foxes from Spain, Bosnia and Herzegovina, Italy, and Portugal. We present here the analysis of uncharacterized nematodes obtained from gastric nodules of a red fox from Switzerland in 1999. The specimens were identified as S. vulpis based on the observation of teeth-like structures in the buccal capsule and a 99.4% sequence identity to S. vulpis DNA from Spain. Phylogenetic analysis demonstrated the clustering of the Swiss sequences in a different group from specimens of other geographical locations. Altogether, this study constitutes the first report of S. vulpis in Switzerland and a report of the oldest specimen of this species in the world. Our findings highlight the widespread distribution of S. vulpis in Europe which may be facilitated by the free-roaming nature of red foxes. [ABSTRACT FROM AUTHOR]

Global warming and displacement of vectors and wild and domestic hosts by humans greatly change host-parasite interactions and parasite transmission rates. Thelazia callipaeda Railliet and Henry, 1910 (Spirurida: Thelaziidae) is a zoonotic parasite rapidly colonizing Europe from its Asian native range. This nematode is vectored by Phortica flies and may cause ocular disorders, such as keratitis and corneal ulcers, in wild mammals, dogs, cats and humans. With the aim of detecting possible threats for the European wildcat (Felis silvestris) in one of its last Mediterranean strongholds, we initiated in 2017 a long-term monitoring program in Sierra Arana (southeastern Spain), which includes exploring its epidemiological and spatial relationships with domestic cats (Felis catus). During routine medical check-ups, we detected ocular nematodes in 3 of 17 (17.6%) wildcats and 1 of 23 (4.3%) domestic cats tested, confirmed to be T. callipaeda by microscopic and molecular analyses. This nematode species was first detected in the study area in 2021. To our knowledge, this is the first case of infection in wildcats in Spain. Through telemetry, we detected little spatial overlap between the home ranges of wildcats and domestic cats, which seems to be sufficient to facilitate the inter-specific transmission of T. callipaeda. Our findings suggest that this pathogen could be a new threat to the wildcat, a locally endangered wild felid. Further eco-epidemiological monitoring and sanitary control of domestic cats will be strongly needed, especially given the ongoing global warming scenario., (© 2023. The Author(s).)

Metathelazia capsulata is a lungworm that inhabit in the bronchi and bronchioles from mammal carnivore species, which life cycle is unknown. M. capsulata-like spirurid nematodes were isolated at necropsy from the respiratory tract of red foxes (Vulpes vulpes) from the Region of Murcia (SE Spain). The main objective of this study was to describe in detail the morphometric features of these nematodes, as well as to report some molecular markers. The principal morphometric difference compared to previous M. capsulata descriptions was the shorter total length for both males and females (6.6 mm and 7.4 mm, respectively). In addition, the mean values of buccal cavity depth and distance between the excretory pore and the anterior end of the nematode were also lower than those previously reported. On the other hand, sequence data of the mitochondrial (COI) and nuclear (rDNA) genes of M. capsulata were described, being the first time that molecular markers are reported for the genus Metathelazia and also for the entire family Pneumospiruridae. Based on data available from GenBank, these results indicate that M. capsulata sequences are closely related to the family Rhabdochonidae, which is assumed to belong to superfamily Thelazioidea, a superfamily including the family Pneumospiruridae, but also suggest the distant relations with the family Thelaziidae. This is the first time that M. capsulata is reported in red fox from Europe. This study provides valuable information for future phylogenetic studies on Metathelazia spp. nematodes and, in general, on species of the family Pneumospiruridae., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Abstract: A telephone survey of farmers was conducted to determine current internal parasite control practices on sheep farms in Spain; the farmers were interviewed by their veterinarians. Anthelmintic choice was largely on veterinary advice and dominated by benzimidazoles and macrocyclic lactones. Anthelmintic rotation was separated into: no rotation (42% of farms); annual rotation (36%); rotate within year (20%); and rotate every second year (2%). The mean annual number of treatments varied subtly by region; ewes and rams 1.6–2.1, replacement lambs 1.7–2.1. Anthelmintics are administered primarily during spring and early summer (47% of treatments), and autumn (41%). Thirty-two percent of farmers introduced sheep to their properties and more than half did not quarantine drench the arrivals. [Copyright &y& Elsevier]

The article focuses on a study about the community structure and feeding preference of nematodes in relation to methane seepage at the Darwin mud volcano (MV) located at the Gulf of Cádiz, Spain. It is said that cold seep sites have few dominant species due to lower species diversity, adding that food sources for seep fauna range from organic matter from chemoautotrophic bacteria to photosynthetic organic matter. Findings show that nematode genus diversity exhibits the lowest levels in the seep sediment core and increases with distance. The study also notes that thiotrophic C forms part of the diet of nematodes up to 10 meters (m) from seepage site.

Paralongidorus litoralis sp. n., a new bisexual species of the genus, is described and illustrated by light microscopy, scanning electron microscopy and molecular studies from specimens collected in a coastal sand dune soil around roots of lentisc (Pistacia lentiscus L.) from Zahara de los Atunes (Cadiz), southern Spain. Paralongidorus litoralis sp. n. is characterised by the large body size (7.5-10.0 mm), a rounded lip region, clearly offset from the body by a collar-like constriction, and bearing a very large stirrup-shaped, amphidial fovea, with conspicuous slit-like aperture, a very long and flexible odontostyle ca 190 μm long, guiding ring located at 35 μm from anterior end, and males with spicules ca 70 μm long. In addition, identification data of a Spanish population of P. paramaximus Heyns, 1965 recovered from sandy soil of a commercial citrus orchard at Alcala de Guadaira (Seville), southern Spain, agree very well with the original description of the species from South Africa. The 18S rRNA and D2 and D3 expansion regions of 28S rRNA gene sequences were obtained for P. litoralis sp. n. and P. paramaximus. Phylogenetic analyses of P. litoralis sp. n. and P. paramaximus rRNA gene sequences and of Longidoridae sequences published in GenBank were done using maximum likelihood and Bayesian inference. In trees generated from the 18S data set Paralongidorus clustered as an external clade from Longidorus, and in trees generated from D2-D3 of 28S dataset Paralongidorus was monophyletic and nested within Longidorus. Maximum likelihood test supported the hypothesis of validity of the Paralongidorus genus. [ABSTRACT FROM AUTHOR]

Host immunocompetence assessed by spleen size and response to phytohaemagglutinin (PHA) injection may give some indications on the control of parasite infection and on host mediation effect, through immunity, on parasite community structure. We investigated the helminth community and immunocompetence of the white-toothed shrew in a small area to test the relationship between immunocompetence and intensity of helminth infection. At the proximate level and if spleen mass and PHA response reflect the level of immunocompetence, we expected that individuals with a large spleen or a high PHA response should harbour a lower parasite load than individuals with relatively small spleen or low PHA response. In addition, we predicted that the structure of the helminth community should be mediated by the host’s immune defence. Spleen mass was linked to helminth infection. Nematodes and cestodes were negatively associated within hosts. PHA response was not related to total helminth intensity of infection but was negatively related to cestode intensity and positively to nematode intensity. This result suggests either a differential modulating effect on immunity by the two groups of worms or the existence of an antagonistic association between nematodes and cestodes mediated by the immune response of the host. [ABSTRACT FROM AUTHOR]

Some of the microorganisms present in urban wastewater, which include intestinal protozoa and nematodes, can be pathogenic. Their (oo)cyst and egg transmissible stages are very resistant to environmental stresses and disinfectants and they are therefore difficult to remove. Thus, they can constitute a health risk if water or sludge obtained in the purification of wastewater is reused for agricultural purposes. In this context, the presence of intestinal protozoa and nematodes were studied in influents, effluents and sludge from five wastewater treatment plants (WWTPs) in the north of Spain by optical microscopy and PCR techniques. The removal efficiency of different wastewater treatments was also compared. The presence of protozoa has increased among the population discharging waste to WWTPs in recent years. Cryptosporidium spp., Giardia duodenalis , Entamoeba spp. and nematodes were detected in all of the WWTPs. Indeed, this is the first report of Entamoeba histolytica and Entamoeba moshkovskii in Spanish WWTPs. The water treatments studied showed different removal efficiencies for each species of intestinal protozoa, with the aerated lagoons providing the best results. (Oo)cysts were also detected in sludge even after aerobic digestion and dehydration. To avoid risks, (oo)cyst viability should be analysed whenever the sludge is to be used as a fertilizer. This study reinforces the necessity of establishing legal limits on the presence of protozoa in WWTP effluents and sludges, especially if reuse is planned. Further studies are necessary for a better understanding of the presence and behaviour of intestinal parasites. Image 10591 • Wastewater treatment plants did not remove intestinal protozoa from water or sludge. • E. histolytica was found after a tertiary treatment based on aerated lagoons. • E. histolytica and E. moshkovskii were detected for the first time in Spanish WWTPs. • Protozoa presence in wastewater reflects their infective occurrence in the population. • (Oo)cyst viability studies are needed for effluents and sludge reuse in agriculture. [ABSTRACT FROM AUTHOR]

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 .

Anisakiasis is a fish-borne parasitic disease caused by the consumption of raw or undercooked fish, as well as cephalopods, contaminated by third instar larvae (L3) of species belonging to the genus Anisakis (Anisakidae). Origanum compactum is a small herbaceous aromatic plant endemic to Spain and Morocco. In Morocco, the plant is used under infusion to treat heart diseases and intestinal pains or as preservative for foodstuffs. This is the first time that the O. compactum essential oil is tested against the parasitic nematode Anisakis simplex. The phytochemical analysis by GC-MS revealed carvacrol (50.3%) and thymol (14.8%) as the major oil constituents. The essential oil and its major constituents carvacrol and thymol were tested against A. simplex L3 larvae isolated from blue whiting fish (Micromesistius poutassou). A. simplex mortality (%) after 24 and 48 h of treatment at 1 μl/ml was 100%, with a low LD 50 compared with other essential oils and extracts, and the penetration in the agar assay was also reduced, if compared with control wells. The oil, as well as its major constituents, demonstrated a dose-dependent larvicidal activity. Inhibition of the enzyme acetylcholinesterase through a colorimetric assay in 96-well plates was used to elucidate the pharmacological mechanism as this enzyme plays a key role in nematodes neuromuscular function. Interestingly, O. compactum essential oil, carvacrol and thymol inhibited the enzyme, confirming that this could be one of the mechanisms involved in the anthelmintic activity. To the best of our knowledge, this is the first time that O. compactum essential oil is reported as a larvicidal agent against A. simplex L3 larvae.