Molecular identification and prevalence of plerocercoid larvae (Cestoda: Diphyllobothriidae) in some commercial fish species from Peru.

Diphyllobothriosis, a fish-borne zoonosis in South America, is mainly caused by the Pacific broad tapeworm Adenocephalus pacificus Nybelin, 1931, a parasite of considerable concern in fishery resources due to its impact on public health. A new diphyllobothrid, Diphyllobothrium sprakeri Hernández-Orts et al. Parasites Vectors 14:219, 2021, was recently described from sea lions from the Pacific Coast, but marine fish acting as intermediate hosts are unknown. The objective of this study was to confirm the presence of plerocercoid larvae of Diphyllobothriidae Lühe, 1910 (Cestoda: Diphyllobothriidea) in nine fish species of commercial importance in Peru. Of a total of 6999 fish (5861 Engraulis ringens, 853 Sciaena deliciosa, 6 Sciaena callaensis, 171 Scomber japonicus, 40 Trachurus murphyi, 40 Ariopsis seemanni, 18 Merluccius peruanus, 5 Sarda chiliensis, and 5 Coryphaena hippurus), 183 were infected with plerocercoid larvae, representing a total prevalence of 2.61% and a mean intensity of 3.2. Based on mtDNA cox1 sequences of 43 plerocercoids, a phylogenetic analysis revealed that 41 belong to A. pacificus and two to D. sprakeri. These findings are first molecular data for D. sprakeri larvae, and the infections of E. ringens and T. murphyi by plerocercoid larvae represent the first records of intermediate/paratenic hosts for this species. Hence, the findings of the current study enhance our understanding of the presence of diphyllobothriid species in commercial fish from the Southeastern Pacific Ocean and their potential impact on seafood safety for local human populations.

Human diphyllobothriasis in Argentina: assessing the epidemiological significance from historical records and reports of new cases.

Between 1898 and 1940, eight human cases of diphyllobothriasis were reported in Argentina, always in recently arrived European immigrants. In 1982, the first autochthonous case was detected, and since then, 33 other autochthonous cases have been reported, totaling 42 cases of human diphyllobothriasis in Argentina before the present study. Our aim is to update the information on diphyllobothriasis in Argentina by identifying specimens from new cases using morphometrical and/or molecular methods. We also aim to assess the epidemiological relevance of this food-borne disease in the country. Anamnestic data were obtained from patients or professionals, along with 26 worms identified using morphometrical (21 samples) and molecular techniques (5 samples). All the patients acquired the infection by consuming freshwater salmonids caught in Andean lakes in Northern Patagonia. Morphometrics and DNA markers of worms were compatible with Dibothriocephalus latus. In total, 68 human cases have been detected in Argentina, 60 of which were autochthonous. The human population living North-western Patagonia, whose lakes are inhabited by salmonids, is increasing. Similarly, the number of other definitive hosts for Dibothriocephalus dendriticus (gulls) and for D. latus (dogs) is also increasing. In addition, salmonid fishing and the habit of consuming home-prepared raw fish dishes are becoming widespread. Therefore, it is to be expected that diphyllobothriasis in Argentina will increase further.

PALEOEPIDEMIOLOGY OF DIPHYLLOBOTHRIOSIS: CULTURAL AND ENVIRONMENTAL FACTORS AFFECTING ADENOCEPHALUS INTENSITY AND PREVALENCE.

Peruvian and Chilean mummies and coprolites provide a source of population-based parasitological information. This is especially true of the fish tapeworm, Adenocephalus pacificus. Our analysis of Chinchorro and Chiribaya mummies and diversified coprolite samples from Chile and Peru show variation in infection. There is a statistically significant difference in prevalence between Chinchorro hunter-gatherer and Chiribaya mixed-subsistence contexts. Furthermore, the most pronounced differences occur between populations within these groups. Chinchorro differences in cemeteries at the same location can be related to El Niño-Southern Oscillation variations. Pronounced prevalence variations between 3 Chiribaya villages within 7 km of each other relate to fish distribution and preparation variation. As with other recent archaeoparasitology studies, eggs-per-gram data exhibit overdispersion.

A possible circulation of a dominant Dibothriocephalus nihonkaiensis haplotype in Japan revealed by molecular analysis of clinical tapeworm samples.

Human diphyllobothriasis, caused by Dibothriocephalus nihonkaiensis, is prevalent globally, especially in regions where raw fish is consumed. Recent molecular diagnostic techniques have made species identification of tapeworm parasites and the determination of genetic variations among parasite populations possible. However, only a few studies done over a decade ago, have reported on the genetic variation among D. nihonkaiensis in Japan. The present study employed PCR-based mitochondrial DNA analysis to specifically detect D. nihonkaiensis from archived clinical samples, and to determine any genetic variation that may exist among the Japanese broad tapeworms from patients of Kanagawa Prefecture, Japan. Target genes were amplified from DNA extracted from the ethanol- or formaldehyde-fixed samples by PCR. Further sequencing and comparative phylogenetic analyses based on mitochondrial COI and ND1 sequences were also performed. In our results, all PCR-amplified and sequenced samples were identified as D. nihonkaiensis. Analysis of COI sequences revealed two haplotype lineages. However, clustering of almost all COI (and ND1) sample sequences into one of the two haplotype clades, together with reference sequences from different countries worldwide, revealed a common haplotype among D. nihonkaiensis samples in our study. Our results suggest a possible presence of a dominant D. nihonkaiensis haplotype, with a global distribution circulating in Japan. Results from this study have the potential to improve the management of clinical cases and establish robust control measures to reduce the burden of human diphyllobothriasis in Japan.

An examination of nervous system revealed unexpected immunoreactivity of both secretory apparatus and excretory canals in plerocercoids of two broad tapeworms (Cestoda: Diphyllobothriidea).

Dibothriocephalus ditremus and Dibothriocephalus latus are diphyllobothriidean tapeworms autochthonous to Europe. Their larval stages (plerocercoids) may seriously alter health of their intermediate fish hosts (D. ditremus) or cause intestinal diphyllobothriosis of the final human host (D. latus). Despite numerous data on the internal structure of broad tapeworms, many aspects of the morphology and physiology related to host­parasite co-existence remain unclear for these 2 species. The main objective of this work was to elucidate functional morphology of the frontal part (scolex) of plerocercoids, which is crucial for their establishment in fish tissues and for an early attachment in final hosts. The whole-mount specimens were labelled with different antibodies and examined by confocal microscope to capture their complex 3-dimensional microanatomy. Both species exhibited similar general pattern of immunofluorescent signal, although some differences were observed. In the nervous system, FMRF amide-like immunoreactivity (IR) occurred in the bi-lobed brain, 2 main nerve cords and surrounding nerve plexuses. Differences between the species were found in the structure of the brain commissures and the size of the sensilla. Synapsin IR examined in D. ditremus occurred mainly around FMRF amide-like IR brain lobes and main cords. The unexpected finding was an occurrence of FMRF amide-like IR in terminal reservoirs of secretory gland ducts and excretory canals, which has not been observed previously in any tapeworm species. This may indicate that secretory/excretory products, which play a key role in host­parasite relationships, are likely to contain FMRF amide-related peptide/s.

[Implementation of a network drug strategy: the case of praziquantel]. / Implementación de una estrategia de medicamentos en red: el caso de praziquantel.

Parasitic infections cause a huge burden of disease and are a current public health problem. The category of emerging or re-emerging disease is influenced by phenomena that occur in today's interconnected world because of globalization, the displacement of people, trade, uncoordinated urbanization and climate change, they have a very important influence on transmission of these diseases. In 2021 there was an increase in the number of patients who have required treatment for diphyllobothriasis in the Los Ríos Region. This article reviews aspects related to integrated Health Service networks to provide access to pharmacological treatments to patients diagnosed with diphyllobothriasis (tapeworm infection), implemented by the Valdivia Health Service Department, in collaboration with the San José de Osorno hospital, primary care centers and private centers in the Los Ríos Region.

Intestinal parasites in the Neolithic population who built Stonehenge (Durrington Walls, 2500 BCE).

Durrington Walls was a large Neolithic settlement in Britain dating around 2500 BCE, located very close to Stonehenge and likely to be the campsite where its builders lived during its main stage of construction. Nineteen coprolites recovered from a midden and associated pits at Durrington Walls were analysed for intestinal parasite eggs using digital light microscopy. Five (26%) contained helminth eggs, 1 with those of fish tapeworm (likely Dibothriocephalus dendriticus) and 4 with those of capillariid nematodes. Analyses of bile acid and sterol from these 5 coprolites show 1 to be of likely human origin and the other 4 to likely derive from dogs. The presence of fish tapeworm reveals that the Neolithic people who gathered to feast at Durrington Walls were at risk of infection from eating raw or undercooked freshwater fish. When the eggs of capillariids are found in the feces of humans or dogs it normally indicates that the internal organs (liver, lung or intestines) of animals with capillariasis have been eaten, and eggs passed through the gut without causing disease. Their presence in multiple coprolites provides new evidence that internal organs of animals were consumed. These novel findings improve our understanding of both parasitic infection and dietary habits associated with this key Neolithic ceremonial site.

Implementación de una estrategia de medicamentos en red: el caso de praziquantel / Implementation of a network drug strategy: the case of praziquantel

Resumen Las infecciones parasitarias provocan una enorme carga de enfermedad y constituyen un problema presente para la salud pública. Las enfermedades emergentes o reemergentes se ven influenciadas por fenómenos del mundo actual interconectado producto de la globalización, el desplazamiento de las personas, el comercio, la urbanización descoordinada y el cambio climático, contribuyendo en la transmisión de estas enfermedades. En el año 2021 hubo un aumento de la cantidad de pacientes que han requerido tratamiento para la difilobotriasis en la Región de los Ríos. Se revisan los aspectos relacionados con las redes integradas de servicios de salud para el acceso al tratamiento farmacológico a pacientes con diagnóstico de difilobotriasis, implementado por la Dirección de Servicio de Salud Valdivia, en colaboración con el hospital San José de Osorno, centros de atención primaria y centros privados de la Región de los Ríos.

Abstract Parasitic infections cause a huge burden of disease and are a current public health problem. The category of emerging or re-emerging disease is influenced by phenomena that occur in today's interconnected world because of globalization, the displacement of people, trade, uncoordinated urbanization and climate change, they have a very important influence on transmission of these diseases. In 2021 there was an increase in the number of patients who have required treatment for diphyllobothriasis in the Los Ríos Region. This article reviews aspects related to integrated Health Service networks to provide access to pharmacological treatments to patients diagnosed with diphyllobothriasis (tapeworm infection), implemented by the Valdivia Health Service Department, in collaboration with the San José de Osorno hospital, primary care centers and private centers in the Los Ríos Region.

Population genetic structure of diphyllobothriid tapeworms (Cestoda: Diphyllobothriidea) parasitising fish in the Baikal Rift Zone.

Tapeworms of the genus Dibothriocephalus are widely distributed throughout the world, and some are agents of human diphyllobothriasis, one of the most important fish-borne zoonoses caused by a cestode parasite. Until now, the population genetic structure of diphyllobothriid tapeworms in the Baikal Rift Zone (BRZ) has remained unexplored. The major aim of this study was to analyse the population genetic structure of D. dendriticus and D. ditremus parasitising fish in the BRZ based on internal transcribed spacer 1 (ITS1) and mitochondrial gene cytochrome oxidase subunit I (cox1) sequences. We found that both species had complex population genetic structures. Each species formed 2 clades (D. dendriticus: Clade 1 & 2; D. ditremus Clade A & B) that differed in genetic diversity. D. dendriticus haplotypes in Clade 1 formed a star-like sub-network with a main haplotype, whereas the haplotypes in Clade 2 formed a diffuse network. We assumed that the complex population genetic structure of D. dendriticus was a consequence of populations evolving under different palaeoecological conditions during the Last Glacial Maximum. In contrast to D. dendriticus, both clades in the D. ditremus samples formed a diffuse network. Our findings revealed hypothetical pathways in the formation of the population genetic structure of diphyllobothriids in the BRZ. On one hand, isolation by distance played an important role; on the other hand, lake recolonisation from refugia and a genetic bottleneck after the end of the Last Glacial Maximum had a possible influence.

Diphyllobothrium stemmacephalum (Cestoda: Diphyllobothriidae) found from a harbor porpoise in northern Japan, with comments on a geographical gap with human infection cases in southern Japan.

Even though the cetacean tapeworm Diphyllobothrium stemmacephalum occurs in both cold and warm waters, human infections and final host occurrences have been confined to temperate areas in and near Japan. We recently obtained a strobila of this cestode that was excreted from a harbor porpoise accidentally caught offshore of Hokkaido of northern Japan. Genetic analysis of 28S rDNA and cox1 genes confirmed that the cestode was D. stemmacephalum. Our finding sets the northernmost record of D. stemmacephalum in the western Pacific, suggesting that the risk of human infections by this parasite in northern Japan deserves further attention.

Gastrointestinal parasites in captive and free-living wild birds in Goiania Zoo / Parasitas gastrointestinais em aves silvestres em cativeiro e de vida livre no Zoológico de Goiânia

Helminths are endoparasites that infect a variety of bird species. Endoparasite infections can cause severe diseases, including kill captive avian hosts and represents a problem to maintenance and conservation. The eggs and larval forms of these parasites are usually eliminated in the host feces. The main interest of this study is to report the occurrence of eggs and oocysts in feces from captive wild birds in Goiânia Zoo and free-living birds in its surroundings. The fecal samples were subjected to parasitological examination to identify the presence of helminths and to classify their eggs based on morphological characteristics. Eggs of nematode parasites (Positive/N) were identified as Ascaridia spp. in Brotogeris chiriri (2/2), Dromaius novaehollandiae (1/1) and Rhea americana (2/2); Ascarididae in Pavo cristatus nigripensis (1/1); Capillaria spp. in Ara chloropterus (1/1) and Penelope jacucaca (1/1); Capillaria plagiaticia in Anodorhynchus hyacinthinus (1/2) and Ara spp. (2/2); Capillaria venusta in Rhamphastus tucanus (1/2); and unidentified nematode eggs from Amazona amazonica (2/2). Eimeria spp. oocyst was founded in feces from A. hyacinthinus (1/2). Considering free-living birds, Diphyllobothrium spp. eggs were founded in Ardea alba (2/2) and Nycticorax nycticorax (2/3). Co-infection of Eustrongylides spp. was founded in A. alba (1/2). This is the first occurrence of: Ascaridia spp. parasitizing B. chiriri; and C. venusta parasitizing R. tucanus. In conclusion, the helminth eggs found in the abovementioned host bird species are consistent with those reported in the literature, and treatment and control protocols were based on their identification.

Helmintos são endoparasitas que infectam uma variedade de espécies de aves. As infecções por endoparasitas podem causar doenças graves, podendo matar aves em cativeiro e representam um problema de manutenção e conservação. Os ovos e as formas larvais desses parasitas são geralmente eliminados nas fezes do hospedeiro. O principal objetivo deste estudo é relatar a ocorrência de ovos e oocistos em fezes de aves silvestres em cativeiro no Zoológico de Goiânia e em aves de vida livre em seu entorno. As amostras fecais foram submetidas a exame parasitológico para identificação da presença de helmintos e classificação dos ovos com base nas características morfológicas. Ovos de nematóides (Positivo/N) foram identificados como Ascaridia spp. em Brotogeris chiriri (2/2), Dromaius novaehollandiae (1/1) e Rhea americana (2/2); Ascarididae em Pavo cristatus nigripensis (1/1); Capillaria spp. em Ara chloropterus (1/1) e Penelope jacucaca (1/1); Capillaria plagiaticia em Anodorhynchus hyacinthinus (1/2) e Ara spp. (2/2); Capillaria venusta em Rhamphastus tucanus (1/2); e ovos de nematóide não identificado de Amazona amazonica (2/2). Oocistos de Eimeria spp. foram encontrados nas fezes de A. hyacinthinus (1/2). Considerando pássaros de vida livre, ovos de Diphyllobothrium spp. foram encontrados em Ardea alba (2/2) e Nycticorax nycticorax (2/3). Coinfecção de Eustrongylides spp. foi encontradada em A. alba (1/2). Esta é a primeira ocorrência de: Ascaridia spp. parasitando B. chiriri; e C. venusta parasitando R. tucanus. Em conclusão, os ovos de helmintos encontrados nas espécies de aves hospedeiras acima mencionadas são consistentes com os relatados na literatura, e os protocolos de tratamento e controle foram baseados em sua identificação.

Ups and downs of infections with the broad fish tapeworm Dibothriocephalus latus in Europe from 1900 to 2020: Part I.

The broad fish tapeworm, Dibothriocephalus latus (Diphyllobothriidea), is the most frequent causative agent of diphyllobothriosis, a fish-borne zoonosis, in Europe. Diphyllobothriosis is characterized by the transmission of D. latus larvae to humans via the consumption of raw, marinated, smoked or inadequately cooked fish products. The most important European foci of diphyllobothriosis have been Fennoscandia, the Baltic region, the Alpine lakes region, the Danube River region, and several endemic regions in Russia. This review provides basic data on the biology, life cycle, host specificity, methods of identification of D. latus, and a detailed summary of its occurrence in intermediate and definitive hosts in Fennoscandia and the Baltic, Alpine, and Danube regions during the last 120 years (1900-2020). Deeper insight into the unique pattern of distribution of D. latus in endemic regions is provided. The numbers of records are associated with several milestones of particular time periods. The first milestone (historical), which influenced studies on D. latus in Europe, was the period during and after World War II (1941-1950). The second milestone (epidemiological) was the decade 1981-1990, when previous massive health campaigns led to a marked decline of diphyllobothriosis in Europe and less published data on D. latus. Based on recent data, the broad fish tapeworm is either absent or present at very low prevalences in Fennoscandia and the Baltic and Danube regions, but the Alpine lakes region represents a continuous ongoing circulation of the parasite in the natural environment and humans.

Marine fish imported from Argentina as source of human diphyllobothriosis in Europe? Ecological evidence from dolphins.

Diphyllobothriosis caused by the Pacific broad tapeworm Adenocephalus pacificus (syn. Diphyllobothrium pacificum) is an emerging parasitic disease reported also from non-endemic areas, including Europe (Spain). The origin of these human cases is unknown but should be related to fresh marine fish imported from endemic areas. In this study, we molecularly confirmed common dolphins Delphinus delphis off Argentina as euparatenic transit hosts of A. pacificus. Preliminary analysis of their stomach content, together with data from previous studies from the Southwest Atlantic, showed that common dolphins feed almost exclusively on schooling Argentine hake Merluccius hubbsi and Argentine anchovy Engraulis anchoita. Therefore, we suggest that Argentine hake and Argentine anchovy may represent the intermediate hosts of A. pacificus in the Southwest Atlantic, but also in Europe to where M. hubbsi is imported on ice (unfrozen).

A regional study of the zoonotic broad tapeworm Dibothriocephalus spp. in Northwestern Patagonia (Argentina): origin of fishes and coastal cities as factors affecting infection in fishes.

Diphyllobothriosis was first recorded in humans in Argentina in 1892 and in introduced salmonids in 1952. The aim of this work is to assess factors influencing the values of prevalence and abundance of plerocercoids in fishes that could increase the risk of transmission of Dibothriocephalus spp. in Andean Patagonian lakes. We analysed two key issues potentially related to the occurrence of tapeworms in fish: the presence of cities on coastlines (as potential sources of eggs to nearby lakes) and the difference between native and exotic fishes in susceptibility to infection. We investigated the probability of finding parasites in fish, the variation in parasite abundance in different environments and the relationship between host length and occurrence of plerocercoids. A total of 3226 fishes (belonging to six autochthonous and four introduced species) were analysed between 2010 and 2019 in eight environments. Plerocercoids were counted, and a subset was determined molecularly to species level. Two species, Dibothriocephalus latus and Dibothriocephalus dendriticus, were identified from both salmonids and native fishes, this being the first molecular confirmation of these tapeworm species parasitizing native South American fishes. Salmonids had higher levels of infection than native fishes, and these levels were higher in aquatic environments with a city on their coastline. Transmission to humans seems to occur mainly through Oncorhynchus mykiss, which showed the highest infection values and is the species most captured by fishers. Based on previous data and the present results, eggs shed by humans, dogs and gulls in cities could be the principal factors in maintaining the life cycle of this parasite in surrounding aquatic environments.

Diphyllobothrium sprakeri n. sp. (Cestoda: Diphyllobothriidae): a hidden broad tapeworm from sea lions off North and South America.

BACKGROUND: The systematic of several marine diphyllobothriid tapeworms of pinnipeds has been revised in recent years. However, 20 species of Diphyllobothrium from phocids and otariids are still recognized as incertae sedis. We describe a new species of Diphyllobothrium from the intestine of California sea lions Zalophus californianus (Lesson) (type-host) and South American sea lions Otaria flavescens (Shaw). METHODS: Zalophus californianus from the Pacific coast of the USA and O. flavescens from Peru and Argentina were screened for parasites. Partial fragments of the large ribosomal subunit gene (lsrDNA) and the cytochrome c oxidase subunit 1 (cox1) mitochondrial gene were amplified for 22 isolates. Properly fixed material from California sea lions was examined using light and scanning electron microscopy. RESULTS: A total of four lsrDNA and 21 cox1 sequences were generated and aligned with published sequences of other diphyllobothriid taxa. Based on cox1 sequences, four diphyllobothriid tapeworms from O. flavescens in Peru were found to be conspecific with Adenocephalus pacificus Nybelin, 1931. The other newly generated sequences fall into a well-supported clade with sequences of a putative new species previously identified as Diphyllobothrium sp. 1. from Z. californianus and O. flavescens. A new species, Diphyllobothrium sprakeri n. sp., is proposed for tapeworms of this clade. CONCLUSIONS: Diphyllobothrium sprakeri n. sp. is the first diphyllobothriid species described from Z. californianus from the Pacific coast of North America, but O. flavescens from Argentina, Chile and Peru was confirmed as an additional host. The present study molecularly confirmed the first coinfection of two diphyllobothriid species in sea lions from the Southern Hemisphere.

Molecular cytogenetic analysis of a triploid population of the human broad tapeworm, Dibothriocephalus latus (Diphyllobothriidea).

The large-sized tapeworm Dibothriocephalus latus is known as the broad or fish-borne cestode of mammals that is capable to infect humans and cause diphyllobothriosis. Recently, molecular data on D. latus has been accumulating in the literature and a complete genome sequence has been published; however, little is known about the karyotype and chromosome architecture. In this study, an in-depth karyological analysis of 2 D. latus specimens was carried out. The plerocercoids originated from a perch caught in subalpine Lake Iseo (Italy) and the tapeworms were reared in hamsters. Both specimens contained cells with a highly variable number of chromosomes ranging from18 to 27. Nevertheless, the largest portion of mitotic figures (47%) showed a number corresponding to the triploid set, 3n = 27. Accordingly, the karyotype of the analyzed specimens consisted of 9 triplets of metacentric chromosomes. Fluorescence in situ hybridization (FISH) with the 18S rDNA probe clearly demonstrated the presence of 3 clusters of hybridization signals on the triplet of chromosome 7, thus confirming the triploid status of the specimens. FISH with a telomeric (TTAGGG)n probe confined hybridization signals exclusively to the terminal chromosomal regions, supporting the earlier findings that this repetitive motif is a conserved feature of tapeworm telomeres.