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.

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.

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.

Epidemiology of Diphyllobothrium nihonkaiense Diphyllobothriasis, Japan, 2001-2016.

We report 958 cases of cestodiasis occurring in Japan during 2001-2016. The predominant pathogen was Diphyllobothrium nihonkaiense tapeworm (n = 825), which caused 86.1% of all cases. The other cestode species involved were Taenia spp. (10.3%), Diplogonoporus balaenopterae (3.3%), and Spirometra spp. (0.2%). We estimated D. nihonkaiense diphyllobothriasis incidence as 52 cases/year. We observed a predominance of cases during March-July, coinciding with the cherry salmon and immature chum salmon fishing season, but cases were present year-round, suggesting that other fish could be involved in transmission to humans. Because of increased salmon trade, increased tourism in Japan, and lack of awareness of the risks associated with eating raw fish, cases of D. nihonkaiense diphyllobothriasis are expected to rise. Therefore, information regarding these concerning parasitic infections and warnings of the potential risks associated with these infections must be disseminated to consumers, food producers, restaurant owners, physicians, and travelers.

Clinical characteristics and epidemiology of intestinal tapeworm infections over the last decade in Tokyo, Japan: A retrospective review.

BACKGROUND: Tapeworm (cestode) infections occur worldwide even in developed countries and globalization has further complicated the epidemiology of such infections. Nonetheless, recent epidemiological data on cestode infections are limited. Our objectives were to elucidate the clinical characteristics and epidemiology of diphyllobothriosis and taeniosis in Tokyo, Japan. METHODOLOGY/PRINCIPAL FINDINGS: We retrospectively reviewed 24 cases of human intestinal cestode infection from January 2006 to December 2015 at a tertiary referral hospital in Tokyo, Japan. The patients included were diagnosed with cestode infection based on morphological and/or molecular identification of expelled proglottids and/or eggs and treated in our hospital. Fifteen and 9 patients were diagnosed with diphyllobothriosis and taeniosis, respectively. The median patient age was 31 years (interquartile range [IQR]: 26-42 years), and 13 (54%) were male. Most of the patients (91.7%) were Japanese. All patients were successfully treated with praziquantel without recurrence. Diphyllobothriosis was caused by Diphyllobothrium nihonkaiense in all patients. Taeniosis was due to infection of Taenia saginata in 8 [88.9%] patients and T. asiatica in 1 [11.1%] patient. All patients with taeniosis were infected outside Japan, as opposed to those with diphyllobothriosis, which were domestic. The source locations of taeniosis were mostly in developing regions. The median duration of the stay of the patients with taeniosis at the respective source location was 1 month (IQR: 1-8). CONCLUSIONS/SIGNIFICANCE: The cestode infection, especially with D. nihonkaiense, has frequently occurred, even in Japanese cities, thereby implicating the probable increase in the prevalence of diphyllobothriosis among travelers, as the number of travelers is expected to increase owing to the Tokyo Olympics/Paralympics in 2020. In addition, medical practitioners should be aware of the importance of providing advice to travelers to endemic countries of taeniosis, including the potential risks of infection and preventive methods for these infections.

Diphyllobothriidea in the north area of the Andean Patagonia: Epidemiology in urban dogs, morphometrical and molecular identification, with comments on wild carnivores.

Diphyllobothriidea are the principal agents of Diphyllobothriosis, a widespread food-borne cestodosis. Accurate identification of the species in samples is therefore crucial for diagnosis and epidemiology in wild and domestic animals, and also humans. We aim to identify at specific level the causative agent, and provide an observational, descriptive, and transversal study of the epidemiology of this zoonosis in urban dogs. Also data on wild carnivores from Northwestern Patagonia are presented. Dog feces were collected in thirteen neighborhoods of varying socioeconomic status, and stools were analyzed by two concentration methods. Adult worms were collected and identified by molecular methods. The population of free-roaming dogs in each neighborhood was estimated, and surveys were conducted at all veterinary clinics registered in the Veterinary Medical College of Bariloche city. A total of 36 wild carnivores road killed or found dead in three National Parks were analyzed. Molecular and morphometric analyses of proglottids and eggs from dogs indicate they are infected with D. latum. Twenty out of 118 dog feces were positive for Diphyllobothrium, from 9 out of 13 neighborhoods, with infection values between 10% and 66%. Percentage of infection was correlated positively with the number of free roaming dogs per block, and with Unsatisfied Basic Needs (UBN %), but not with distance to nearest water body. Infection by D. latum in dogs is widely distributed throughout the city. Not all local veterinarians know the occurrence of Diphyllobothriosis in the dogs of the city, and it is evident that this zoonosis is underdiagnosed in relation to the percentage of infection found in this study. None of the analyzed wild carnivores were positive for Diphyllobothrium.

[(THE CURRENT SITUATION OF DIPHYLLOBOTHRIASIS: EPIDEMIOLOGY AND EPIDEMIOLOGICAL SURVEILLANCE)].

The paper analyzes the etiology of diphyllobothriasis pathogens and their distribution in the world. It estimates the infection rate of the population with tapeworms, the value of different fish species and procedures for their cooking as factors for transmission of Diphyllobothrium to the population and the risk for infection with tapeworms through infected foods. Epidemiological surveillance measures in different (synanthropic and natural) foci of diphyllobothriasis and with various types of pathogens are considered.

Prevalence of Diphyllobothrium latum (Cestoda: Diphyllobothriidae) plerocercoids in fish species from four Italian lakes and risk for the consumers.

In recent years there has been a re-emergence of diphyllobothriasis by Diphyllobothrium latum (Cestoda: Diphyllobothriidae) in Italy, France and Switzerland, where in the past this fish-borne zoonosis was widespread and then virtually disappeared. A change in eating habits such as the consumption of raw/undercooked freshwater fish, has led to an increased risk for consumers of ingesting infective larvae of D. latum. A survey on the factors responsible for the re-emergence of human diphyllobothriasis in Italy was carried out from March 2013 to December 2014. The aim of this study was to assess the diffusion of D. latum plerocercoids in the fish populations of the sub-alpine lakes of Maggiore, Como, Iseo and Garda, updating the scarce historical data and assessing a preliminary "risk level" of the lacustrine environments and fish species under investigation. A total of 2228 fish belonging to 5 species, 690 from Lake Maggiore, 500 from Lake Como, 655 from Lake Iseo and 383 from Lake Garda were submitted to parasitological examination. The presence of D. latum plerocercoid larvae was detected in 6.6%, 25.4% and 7.6% of perch (Perca fluviatilis) from Lakes Maggiore, Como and Iseo respectively. The parasite was also present in pike (Esox lucius) with prevalence values ranging from 71.4 to 84.2% and in 3.6-3.8% of burbot (Lota lota) from Lakes Iseo and Como. Fish from Lake Garda were negative as well as sampled whitefish (Coregonus lavaretus) and shad (Alosa fallax lacustris). The results of this survey showed a widespread presence of D. latum plerocercoid larvae in Maggiore, Como and Iseo fish populations. Urban fecal contamination of water is still a key issue to be resolved, together with the improvement of communication with consumers regarding the best dietary habits and the most effective processes of parasite inactivation, required for the consumption of raw/undercooked fish caught in high-risk areas.

Effects of fish species composition on Diphyllobothrium spp. infections in brown trout - is three-spined stickleback a key species?

Subarctic populations of brown trout (Salmo trutta) are often heavily infected with cestodes of the genus Diphyllobothrium, assumedly because of their piscivorous behaviour. This study explores possible associations between availability of fish prey and Diphyllobothrium spp. infections in lacustrine trout populations. Trout in (i) allopatry (group T); (ii) sympatry with Arctic charr (Salvelinus alpinus) (group TC); and (iii) sympatry with charr and three-spined stickleback (Gasterosteus aculeatus) (group TCS) were contrasted. Mean abundance and intensity of Diphyllobothrium spp. were higher in group TCS compared to groups TC and T. Prevalence, however, was similarly higher in groups TCS and TC compared to group T. Zero-altered negative binomial modelling identified the lowest probability of infection in group T and similar probabilities of infection in groups TC and TCS, whereas the highest intensity was predicted in group TCS. The most infected trout were from the group co-occurring with stickleback (TCS), possibly due to a higher availability of fish prey. In conclusion, our study demonstrates elevated Diphyllobothrium spp. infections in lacustrine trout populations where fish prey are available and suggests that highly available and easily caught stickleback prey may play a key role in the transmission of Diphyllobothrium spp. parasite larvae.

Pacific Broad Tapeworm Adenocephalus pacificus as a Causative Agent of Globally Reemerging Diphyllobothriosis.

The Pacific broad tapeworm Adenocephalus pacificus (syn. Diphyllobothrium pacificum) is the causative agent of the third most common fish-borne cestodosis among humans. Although most of the nearly 1,000 cases among humans have been reported in South America (Peru, Chile, and Ecuador), cases recently imported to Europe demonstrate the potential for spread of this tapeworm throughout the world as a result of global trade of fresh or chilled marine fish and travel or migration of humans. We provide a comprehensive survey of human cases of infection with this zoonotic parasite, summarize the history of this re-emerging disease, and identify marine fish species that may serve as a source of human infection when eaten raw or undercooked.

Diphyllobothrium sp. en Canis familiares de la región del subtrópico argentino (Puerto Iguazú, Misiones) / Diphyllobothrium sp. in Canis familiaris from the subtropical area of Argentina (Puerto Iguazú, Misiones)

En este trabajo se informa el hallazgo de huevos de Diphyllobothrium sp. en ejemplares de Canis familiaris (perro doméstico) de Puerto Iguazú, una ciudad subtropical de la provincia de Misiones, Argentina. Durante 2013, en el marco de un relevamiento de la fauna parasitológica de los perros de Puerto Iguazú, se detectaron dos casos positivos en la búsqueda de huevos de Diphyllobothrium sp. La materia fecal de los perros fue recolectada en frascos con formol al 10% y procesada mediante las técnicas de sedimentación de Telemann y de flotación de Sheather. Dado que Misiones no forma parte de la zona endémica de difilobotriasis y considerando, además, su ubicación fronteriza, este hallazgo reviste gran importancia para la salud pública. Se señala la necesidad de actualizar el estado de conocimiento sobre el ciclo de vida de estos parásitos identificando el rango de hospederos intermediarios y definitivos, su potencial zoonótico y la situación epidemiológica en áreas no endémicas

This paper reports the first finding of Diphyllobothrium sp. eggs in Canis familiaris (domestic dog) from Puerto Iguazú, a subtropical city of Misiones province, Argentina. In 2013, two positive cases of Diphyllobothrium sp. eggs were detected during an annual parasitological survey of dogs. Dog feces were collected in vials containing 10% formalin and processed using Telemann's sedimentation and Sheather's flotation techniques. The two cases were detected in rural areas of the municipality. Since Misiones is not a part of the endemic area of diphyllobothriasis and given the fact that it is located in the three-border area of Argentina, Brazil and Paraguay, we consider this finding of great importance to public health. We stress the need for updating the current knowledge about the life cycle of these parasites considering the range of intermediate and definitive hosts, their zoonotic potential, and the epidemiological situation in non-endemic areas

Diphyllobothrium sp. in Canis familiaris from the subtropical area of Argentina (Puerto Iguazú, Misiones).

This paper reports the first finding of Diphyllobothrium sp. eggs in Canis familiaris (domestic dog) from Puerto Iguazú, a subtropical city of Misiones province, Argentina. In 2013, two positive cases of Diphyllobothrium sp. eggs were detected during an annual parasitological survey of dogs. Dog feces were collected in vials containing 10% formalin and processed using Telemann's sedimentation and Sheather's flotation techniques. The two cases were detected in rural areas of the municipality. Since Misiones is not a part of the endemic area of diphyllobothriasis and given the fact that it is located in the three-border area of Argentina, Brazil and Paraguay, we consider this finding of great importance to public health. We stress the need for updating the current knowledge about the life cycle of these parasites considering the range of intermediate and definitive hosts, their zoonotic potential, and the epidemiological situation in non-endemic areas.

Diphyllobothrium nihonkaiense: wide egg size variation in 32 molecularly confirmed adult specimens from Korea.

The eggs of Diphyllobothrium nihonkaiense were reported to be smaller than those of the classical Diphyllobothrium latum in general. However, verification using a large number of adult tapeworms is required. We assessed the egg size variation in 32 adult specimens of D. nihonkaiense recovered from Korean patients in 1975-2014. The diagnosis of individual specimens was based on analysis of the mitochondrial cytochrome c oxidase 1 gene sequence. Uterine eggs (n = 10) were obtained from each specimen, and their length and width were measured by micrometry. The results indicated that the egg size of D. nihonkaiense (total number of eggs measured, 320) was widely variable according to individual specimens, 54-76 µm long (mean 64) and 35-58 µm wide (mean 45), with a length-width ratio of 1.32-1.70 (mean 1.46). The worm showing the smallest egg size had a length range of 54-62 µm, whereas the one showing the largest egg size had a length range of 68-76 µm. The two ranges did not overlap, and a similar pattern was observed for the egg width. Mapping of each egg size (n = 320) showed a wide variation in length and width. The widely variable egg size of D. nihonkaiense cannot be used for specific diagnosis of diphyllobothriid tapeworm infections in human patients.

The influence of changing prey availability on the prevalence of Diphyllobothrium in river otters from Yellowstone National Park.

Parasite prevalence in predatory mammals is influenced by numerous factors including diet, sex, season, and habitat. We examined the effect of such factors on the prevalence of gastrointestinal helminths in North American river otters ( Lontra canadensis ) in Yellowstone Lake, Yellowstone National Park. Otters in this ecosystem have recently experienced a decline in their main prey, Yellowstone cutthroat trout ( Oncorhynchus clarkii bouvieri), and have, in turn, increased consumption of alternative foods. Helminths were found in 13.2% of otter fecal samples. The dominant parasite was a Diphyllobothrium sp. ( Diphyllobothrium ditremum or Diphyllobothrium dendriticum ), a cestode acquired from cutthroat trout. Truttaedacnitis truttae and Contracaecum spp. nematodes were incidental parasites in otter feces, and acanthocephalan eggs were found in 1 sample. The prevalence of trout remains and a Diphyllobothrium sp. in otter feces was higher during the cutthroat trout spawning season. A Diphyllobothrium sp. was more prevalent in the feces of female otters. There was no relationship between annual declines in the frequency of trout in otter feces and prevalence of parasites. Helminth prevalence and species richness in Yellowstone otters was low and likely reflects low predator densities and few intermediate hosts. This is the first study reporting the helminth fauna of river otters in the Greater Yellowstone Area and confirms the otter as a definitive host for Diphyllobothrium sp. in this region.