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.

Eggs as a Suitable Tool for Species Diagnosis of Causative Agents of Human Diphyllobothriosis (Cestoda).

BACKGROUND: Tapeworms of the order Diphyllobothriidea are parasites of tetrapods and several species may infect man and cause neglected human disease called diphyllobothriosis. Identification of human-infecting diphyllobothriid cestodes is difficult because of their morphological uniformity, which concerns also their eggs in stool samples. METHODS: In the present study, we analysed by far the largest dataset of more than 2,000 eggs of 8 species of diphyllobothriid cestodes that may infect humans, including the most frequent human parasites Diphyllobothrium latum, D. nihonkaiense and Adenocephalus pacificus (syn. Diphyllobothrium pacificum). Size (length, width and length/width ratio) and the surface of the egg shell from naturally and experimentally infected hosts were studied using light and scanning electron microscopy. RESULTS: A high degree of intraspecific and host-related size variability has been detected, but combination of morphometrical and ultrastructural data made it possible to distinguish all of the studied species, including otherwise quite similar eggs of the 3 most common species infecting man, i.e. D. latum, D. nihonkaiense and D. dendriticum. The surface of all marine species is covered by numerous deep pits with species-specific density, whereas the surface of freshwater species is smooth or with isolated shallow hollows or wrinkles.

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.

First study of vitellogenesis of the broad fish tapeworm Diphyllobothrium latum (Cestoda, Diphyllobothriidea), a human parasite with extreme fecundity.

In the present study, the process of vitellogenesis of one of the most prolific organisms, the broad tapeworm, Diphyllobothrium latum, the causative agent of human diphyllobothriosis, was studied for the first time using transmission electron microscopy. Cytochemical staining with periodic acid-thiosemicarbazide-silver proteinate for detection of glycogen was applied. Starting from the periphery toward the center of the vitelline follicle four stages of vitellocytes are differentiated: immature vitellocytes, early maturing vitellocytes, advanced maturing and mature vitellocytes. Differentiation into mature vitellocytes involves the formation of shell globule clusters containing shell globules, large amount of saturated lipid droplets and glycogen. A peculiar ultrastructural feature of D. latum vitellogenesis is the presence of lamellar bodies in the cytoplasm of mature vitellocytes. This feature is similar to that present in the closely related caryophyllideans and spathebothriideans. Despite the great similarity observed in the embryonic development of diphylobothriideans, caryophyllideans and spathebothriideans, and the fact that their vitellocytes share a feature not reported from other cestode groups, there are substantial differences in the morphology of vitelline clusters, types, amount and localization of their nutritive reserves.

GABA in the nervous system of the cestodes Diphyllobothrium dendriticum (Diphyllobothriidea) and Caryophyllaeus laticeps (Caryophyllidea), with comparative analysis of muscle innervation.

The nervous system (NS) of the cestodes Diphyllobothrium dendriticum (Diphyllobothriidea) and Caryophyllaeus laticeps (Caryophyllidea) was investigated using immunocytochemistry. The GABA neurotransmitter was identified in the NS of both species; GABAergic neurons were detected in the main nerve cords (MC). GABA-like immunoreactive neurons were predominantly unipolar and exhibited more intensive immunoreactivity in the neurite than in the perikaryon. In C. laticeps , GABA-like immunoreactive somas are located in both the MCs and peripheral NS near the longitudinal muscles. The innervation of the body musculature was studied using a combination of antibodies against GABA, serotonin (5-HT), and FMRFamide and with complementary staining of F-actin. In both species, the location of GABAergic neurites is associated with all muscle layers including the subtegumental, longitudinal, transverse, and dorsoventral muscles. The cytomorphology of 5-HTergic motoneurons in the MCs of both species is described and differences in muscle innervation between D. dendriticum and C. laticeps are demonstrated. No evidence for co-localization of GABA with 5-HT or FMRFamide neurotransmitters at any particular neuron was found. Neuropiles in MCs and peripheral NS had separate sets of immunoreactive neurites. A functional role for GABA in muscle innervation is discussed.

Tapeworm Diphyllobothrium dendriticum (Cestoda)--neglected or emerging human parasite?

BACKGROUND: A total number of 14 valid species of Diphyllobothrium tapeworms have been described in literature to be capable of causing diphyllobothriosis, with D. latum being the major causative agent of all human infections. However, recent data indicate that some of these infections, especially when diagnosed solely on the basis of morphology, have been identified with this causative agent incorrectly, confusing other Diphyllobothrium species with D. latum. Another widely distributed species, D. dendriticum, has never been considered as a frequent parasite of man, even though it is found commonly throughout arctic and subarctic regions parasitizing piscivorous birds and mammals. Recent cases of Europeans infected with this cestode called into question the actual geographic distribution of this tapeworm, largely ignored by medical parasitologists. METHODOLOGY AND RESULTS: On the basis of revision of more than 900 available references and a description and revision of recent European human cases using morphological and molecular (cox1) data supplemented by newly characterized D. dendriticum sequences, we updated the current knowledge of the life-cycle, geographic distribution, epidemiological status, and molecular diagnostics of this emerging causal agent of zoonotic disease of man. CONCLUSIONS: The tapeworm D. dendriticum represents an example of a previously neglected, probably underdiagnosed parasite of man with a potential to spread globally. Recent cases of diphyllobothriosis caused by D. dendriticum in Europe (Netherlands, Switzerland and Czech Republic), where the parasite has not been reported previously, point out that causative agents of diphyllobothriosis and other zoonoses can be imported throughout the world. Molecular tools should be used for specific and reliable parasite diagnostics, and also rare or non-native species should be considered. This will considerably help improve our knowledge of the distribution and epidemiology of these human parasites.

Update on the human broad tapeworm (genus diphyllobothrium), including clinical relevance.

SUMMARY: Tapeworms (Cestoda) continue to be an important cause of morbidity in humans worldwide. Diphyllobothriosis, a human disease caused by tapeworms of the genus Diphyllobothrium, is the most important fish-borne zoonosis caused by a cestode parasite. Up to 20 million humans are estimated to be infected worldwide. Besides humans, definitive hosts of Diphyllobothrium include piscivorous birds and mammals, which represent a significant zoonotic reservoir. The second intermediate hosts include both freshwater and marine fish, especially anadromous species such as salmonids. The zoonosis occurs most commonly in countries where the consumption of raw or marinated fish is a frequent practice. Due to the increasing popularity of dishes utilizing uncooked fish, numerous cases of human infections have appeared recently, even in the most developed countries. As many as 14 valid species of Diphyllobothrium can cause human diphyllobothriosis, with D. latum and D. nihonkaiense being the most important pathogens. In this paper, all taxa from humans reported are reviewed, with brief information on their life history and their current distribution. Data on diagnostics, epidemiology, clinical relevance, and control of the disease are also summarized. The importance of reliable identification of human-infecting species with molecular tools (sequences of mitochondrial genes) as well as the necessity of epidemiological studies aimed at determining the sources of infections are pointed out.

Spermatological characters in Diphyllobothrium latum (Cestoda, Pseudophyllidea).

Spermiogenesis and the ultrastructural features of the spermatozoon of Diphyllobothrium latum (Cestoda, Pseudophyllidea) are described using transmission electron microscopy. Spermiogenesis is characterized by the development of two flagella of unequal length that grow asynchronously. When the first growing flagellum starts to rotate, the second one develops. Flagellar rotation is thus asymmetric and asynchronic. It is followed by proximodistal fusion with the median cytoplasmic process. Electron-dense material is present in the apical region of the zone of differentiation in the early stages of spermiogenesis. The intercentriolar body consists of seven plates: three are electron-dense. Four attachment zones occur in the median cytoplasmic process. An atypical arrangement of striated roots was occasionally observed. The mature spermatozoon possesses two axonemes of 9 + "1" trepaxonematan pattern, nucleus, cortical microtubules, electron-dense granules, and lacks mitochondria. The ultrastructure of the anterior extremity of the spermatozoon in D. latum clearly differs from that in the bothriocephalid pseudophyllideans, mainly in the absence of a crested body and a ring of electron-dense tubular structures. The spermatological data support the assumption that the order Pseudophyllidea is formed by two unrelated clades, "Bothriocephalidea" and "Diphyllobothriidea."

Identification of the copepod intermediate host of the introduced broad fish tapeworm, Diphyllobothrium latum, in Southern Chile.

The broad fish tapeworm, Diphyllobothrium latum, is an exotic species in both Chile and Argentina, and until now, its copepod host has remained unknown in South American waters. The objective of this study was to identify calanoid copepod species that may be intermediate hosts for D. latum in Lake Panguipulli, Chile. In this lake, the highest levels of infection by this tapeworm occur in the introduced rainbow trout, Oncorhynchus mykiss. Of the 2 calanoid copepods found in Lake Panguipulli, Diaptomus diabolicus and Boeckella gracilipes, only D. diabolicus became infected on experimental exposure to coracidia. Prevalence (mean intensity) of experimental infection in adult copepods was 73.2% (2.8 procercoids per host). Diaptomus diabolicus has been demonstrated to be a new intermediate host; this is the first record of a copepod host for D. latum in South America.

Diphyllobothriosis--experimental study.

For the first time the agent of diphyllobothriosis Lake Baikal is the cestoda D. dendriticum establishted by T.P. Chizhova and P.G. Gofman-Kadoshnikov (1962). D. dendriticum reaches maturity in human intestine, but it is considered, that the relationships between the parasite and human are facultative property (Klebonovski 1985). The results of the investigation of the diet of sea-gull nestlings (control-uninfected and experimental-infected birds with cestoda groups) and stress influence (starvation) on the mass loss in experimentally and naturally infected ones with different duration conditions adaptation of experiments are discussed in this article.

The NADPH-diaphorase staining reaction in relation to the aminergic and peptidergic nervous system and the musculature of adult Diphyllobothrium dentriticum.

A new category of signalling molecules--transmitter gases--has appeared. Nitric oxide (NO) is generated by nitric oxide synthase (NOS) and diffuses as a short-lived transcellular messenger through the plasma membrane. NADPH-diaphorase (NADPH-d) is a marker enzyme for NO-producing neurons. In this study the pattern of NADPH-d stained neurons in Diphyllobothrium dendriticum is described and compared to the pattern of aminergic and peptidergic neuronal elements and to that of the musculature stained with TRITC-labelled phalloidin. NADPH-d staining was observed in neurons in the bilobed brain and along the 2 main nerve cords and in nerve fibres close to the body musculature and the musculature of the reproductive ducts, the walls of the testicular follicles and in sensory endings in the tegument. The NADPH-d staining reaction and the 5-HT or FMRFamide immunoreactivities occur in separate sets of neurons.

[Taenia sp and other intestinal cestode infections in individuals from public outpatient clinics and hospitals from the northern section of Santiago, Chile (1985-1994)]. / Infecciones por Taenia sp. y otros cestodos intestinales en pacientes de consultorios y hospitales públicos del sector norte de Santiago, Chile. 1985-1994.

During 1985-1994 period, 70,642 parasitological stool examinations were performed. The exam was made to each patient which presenting gastrointestinal symptoms or nutritional problems, specially in children. 59,960 out of the 70,642 examinations corresponded to 1 month--15 years old children, 4,280 to 16-30 years old individuals and 6.402 to up to 30 years old adults. The infection rates (x 100,000) were: Taenia sp., 86; Hymenolepis nana, 1,165; Diphyllobotrium sp., 4 and Hymenolepis diminuta, 1. In this period, 11 cases of infection by Taenia saginata were detected, and none Taenia sodium infection. Although infections rates by intestinal cesdodes were higher in males than females, in the Taenia sp. infection the difference was not statistical significative. Médico-veterinary control of animals slaughtered in abbatoirs and the improvement of sanitary conditions in the last two decades have contributed to the control of most of human intestinal cestodiases.

El ciclo evolutivo experimental de Diphyllobothrium erinaceieuropei en Paracyclops fimbriatus, larvas de Bufo arenarum y caninos / Experimental evolutive cycle of Diphyllobothrium erinaceieuropei in Paracyclops fimbriatus, tadpoles of Bufo arenarum and dogs

El objetivo del trabajo fue lograr la reproducción experimental del ciclo evolutivo de Diphyllobothrium erinaceieuropei Rudolphi 1819 (Cestoda, Pseudophyllidea) con la intervención de Paracyclops fimbriatus y larvas de Bufo arenarum como hospedadores intermediarios y caninos como hospedadores definitivos. Los huevos del parásito se obtuvieron de heces de caninos infectados naturalmente y se conservaron refrigerados en agua. Se incubaron 7 días a 25-C para que desarrollaran los coracidios y se pusieron en recipientes que contenían a los copépodos mencionados. Al cabo de 12 días a 22,6-C (promedio) se hallaron procercoides maduros en ellos y se agregaron 10 renacuajos de Bufo arenarum. Estos se examinaron por disección 22, 23, 61 y 107 días después, hallándose en todos 1 o más plerocercoides (Temperatura promedio: 24,9-C. El día 23, de 6 renacuajos se obtuvieron 49 plerocercoides, de los cuales se administraron 28, por vía oral, a una perra. El día 107,3 de 11 plerocercoides obtenidos de un renacuajo se le dieron a otra perra por la misma vía. Se hallaron huevos del cestode en las heces del primer canino a partir del día 22 posterior a la infección (p.i.) y a los 30 días p.i., segmentos de estróbila. En el segundo canino se hallaron huevos a los 30 días p.i.

[Experimental evolutive cycle of Diphyllobothrium erinaceieuropei in Paracyclops fimbriatus, tadpoles of Bufo arenarum and dogs]. / El ciclo evolutivo experimental de Diphyllobothrium erinaceieuropei en Paracyclops fimbriatus, larvas de Bufo arenarum y caninos.

Experiments were performed in order to develop the life cycle of Diphyllobothrium erinaceieuropei Rudolphi 1819 (Cestoda, Pseudophyllidea) in Paracyclops fimbriatus and Bufo arenarum as intermediate hosts and dogs as definitive hosts. The eggs of Diphyllobothrium erinaceieuropei from faeces of naturally infected dogs were kept refrigered, in water. In order to obtain coracidiums they were incubated at 25 degrees C, and then were placed in a flask which contained Paracyclops fimbriatus. The copepods were observed to be infected with procercoids 12 days after, (mean temperature 22.6 degrees C) and then, ten tadpoles of Bufo arenarum were put into the same flask. The tadpoles were examined 22, 23, 61 and 107 days later, finding plerocercoids in all them (mean temperature 24.9 degrees C). On the 23rd day, 49 plerocercoids were removed from 6 tadpoles and 28 of them were fed to a bitch. On the 107th day, 11 plerocercoids were recovered from a dead tadpole and 3 of them were fed to another bitch. In the faeces of the first bitch there were observed the eggs of Diphyllobothrium erinaceieuropei 22 days post infection (d.p.i.) and part of the strobila 30 d.p.i.. In the faeces of the second bitch the eggs were found 30 d.p.i..

[The survival of Diphyllobothrium dendriticum pleurocercoids during the processing of fish and sanitary helminthologic evaluation of the product]. / Vyzhivaemost' plerotserkoidov Diphyllobothrium dendriticum pri pererabotke ryby i sanitarno-gel'mintologicheskaia otsenka produktsii.

Diphyllobothrium dendriticum plerocercoids' survival in salt water solutions of various concentrations and at various temperature was assessed. Conditions of salt treatment and freezing of the Baikal salmon assuring total release of fish from D. dendriticum plerocercoids are determined. The authors suggested differentiation in sanitary helminthological assessment of fish invasion rate by D. latum and D. dendriticum plerocercoids.

[Cestode egg production dynamics and fertility in Diphyllobothrium dendriticum (Cestoda, Pseudophyllidea)]. / Dinamika iaitseproduktsii i plodovitost' tsestody Diphyllobothrium dendriticum (Cestoda, Pseudophyllidea).

Dynamics of egg production of the tapeworm Diphyllobothrium dendriticum has been estimated experimentally in nestlings of the herring gull Larus argentatus per day and per reproductive period. The number of eggs in strobiles has been estimated for the maturation period. Mean egg production of the tapeworm per day (10.43 +/- 2.72 mln) is two orders of magnitude as high as the maximum number of eggs in a mature strobile (0.206 +/- 0.007). It is proposed to estimate coefficient of reproduction intensity as the ratio of egg output per unit of time and maximum egg number in a mature strobile.

Dynamics of egg production of the cestode Diphyllobothrium dendriticum (Nitzsch, 1824) (Cestoda: Pseudophyllidea) and the concept of fecundity in helminths.

The dynamics of egg production of the tapeworm D. dendriticum has been estimated experimentally in nestlings of the herring gull Larus argentatus per day and per reproductive period. Numbers of eggs in strobilae have been estimated for the maturation period. Mean egg production, of one tapeworm per day (10.42 +/- 2.72 mln) is two orders of magnitude as high as the maximal number of eggs in a mature strobila (0.206 +/- 0.007). It is proposed to estimate the coefficient of reproduction intensity as the ratio of egg output per unit of time and maximum egg numbers in a mature strobila.