We investigated the risk of diphyllobothriasis from ingestion of wild Pacific salmon in Japan by surveying Diphyllobothrium plerocercoids in 182 salmon samples obtained from Japan. The plerocercoids were not detected in chum salmon (Oncorhynchus keta) (0/26), called Akizake in Japan, caught between September and November. However, the detection rate of plerocercoids in chum salmon, called Tokishirazu in Japan, caught between early April and June, was 51.1% (24/47) with an average of two plerocercoid larvae per fish. The detection rates of cherry salmon (Oncorhynchus masou) and pink salmon (Oncorhynchus gorbuscha) were 12.2% (10/82) and 18.5% (5/27), respectively, and the average number of plerocercoids per fish was 0.45 (37 larvae/82 fishes) and 0.22 larvae (6 larvae/27 fishes), respectively. Plerocercoids isolated from O. keta and O. masou were identified as Diphyllobothrium nihonkaiense on the basis of molecular analysis of the cox1 and nad3 genes. Moreover, four tapeworms (three from O. keta and one from O. masou) were obtained by infecting golden hamsters with plerocercoids. The morphological features of these tapeworms were similar to those of D. nihonkaiense isolated from humans. Therefore, we think that O. keta and not O. masou is the most important source of plerocercoid infections in Japan.

Despite the advances of molecular tools, new nematode species are still described mainly based on morphological characters. Parapharyngodon and Thelandros are two genera of oxyurids with unclear related taxonomic histories. Here we use morphological characters (linear measurements and categorical variables) and genetic information (18S rRNA, 28S rRNA and COI partial gene sequences) to confirm the relationships between representatives of these two genera and to determine whether they can be discriminated morphologically. Genetic results confirm the existence of two main clades, mostly congruent with Parapharyngodon and Thelandros genera but with several discordances. Thelandros is polyphyletic, with two of the species analysed (T. filiformis and T. tinerfensis) being part of the Thelandros clade, but with a third one (T. galloti) falling within the Parapharyngodon clade. Regarding the Parapharyngodon clade, P. cubensis, P. scleratus and Parapharyngodon sp. from Mexico form congruent lineages, while most P. echinatus samples cluster in another group, with one exception. Interestingly, P. micipsae samples are scattered across the Parapharyngodon clade, suggesting that they were misidentified or rather represent alternative morphotypes of other species. Morphological analysis identified the length of the tail, number of caudal papillae, position of the nerve ring, presence of caudal alae and length of the lateral alae as reliable characters to distinguish between Parapharyngodon and Thelandros genetic clades. Our study highlights the current taxonomic inconsistency in these groups, mainly derived from the exclusive use of morphological data. As such, we advocate for the routine implementation of molecular data in nematode taxonomic studies.

The current study was designed to increase the spermatological ultrastructural database on Digenea, thus contributing to the future establishment of phylogenetic relationships within this group based on ultrastructural characteristics of both spermiogenesis and spermatozoa. Spermiogenesis in Elstia stossichianum begins with the formation of a differentiation zone containing two centrioles, two striated rootlets, a nucleus, several mitochondria and an intercentriolar body constituted by seven electron-dense layers. Each centriole develops into a free flagellum growing orthogonally to the median cytoplasmic process. Proximo-distal fusion of the flagella with the median cytoplasmic process occurs after flagellar rotation. Both nucleus and mitochondria migrate before the complete proximo-distal fusion of both flagella with the median cytoplasmic process. Finally, the constriction of the ring of arched membranes gives rise to the young spermatozoon. The mature spermatozoon of E. stossichianum exhibits two axonemes of different length of the 9+‘1’ trepaxonematan pattern, a nucleus, a mitochondrion, two bundles of parallel cortical microtubules, external ornamentation of the plasma membrane, a lateral expansion, spine-like bodies, cytoplasmic ornamented buttons and granules of glycogen. The formation of cytoplasmic ornamented buttons during the final stages of spermiogenesis is reported here for the first time.

Free-living nematodes commonly occur in cultures of fæces, and as Looss (1911) showed years ago they have frequently been mistaken for the developmental stages of parasitic nematodes. For this reason it is important that detailed knowledge of the morphology of such free-living forms should be available. With this end in view the following illustrated account of what appears to be a new genus of nematode has' been prepared. The worms occurred originally in a culture of the fæces of a brown rat, Rattus norvegicus, made for the study of the free-living stages of the Strongyloides of the rat; and as after detailed study it has been found impossible to find any existing description of nematodes which agree with the structural appearances shown by the worms, it has been decided to create a new genus for their reception.

In the present paper, we analyse the effect of a primary infection of ICR mice with Echinostoma caproni (Trematoda: Echinostomatidae) on the generation of resistance against homologous challenge infections. In ICR mice, E. caproni induces chronic infections concomitantly with strong responses characterized by the development of T-helper 1 (Th1)-type local immune responses with elevated levels of local interferon-gamma (IFN-γ) and inflammatory and antibody responses. Here, the effect of the response generated against a primary infection with E. caproni in the generation of resistance against subsequent homologous infections was analysed. For this purpose, ICR mice were challenged with metacercariae of E. caproni and the results obtained showed that primary infection induces partial resistance against subsequent homologous infections in ICR mice. This resistance was expressed as a reduced rate of infection, worm recovery and worm size, indicating that primary infection induces changes in the host, making a hostile environment for the development of the parasite.

Experimental infections of Lymnaea glabra (two populations) with Fasciola hepatica were carried out during seven successive snail generations, to determine if prevalence and intensity of snail infection increased over time through descendants of snails already infected with F. hepatica. Controls were descendants coming from uninfected parents and infected according to the same protocol. No larval forms were found in the bodies of control snails coming from uninfected parents. In contrast, prevalence and intensity of F. hepatica infection in snails originating from infected parents progressively increased from the F2 or F3 to the F6 generation of L. glabra. In another experiment carried out with the F7 generations of L. glabra and a single generation of Galba truncatula (as controls), the prevalence of F. hepatica infection and the total number of cercariae were lower in L. glabra (without significant differences between both populations). If the number of cercariae shed by infected snails was compared to overall cercarial production noted in snails containing cercariae but dying without emission, the percentage was greater in G. truncatula (69% instead of 52–54% in L. glabra). Even if most characteristics of F. hepatica infection were lower in L. glabra, prevalence and intensity of parasite infection increased with snail generation when tested snails came from infected parents. This mode of snail infection with F. hepatica suggests an explanation for cases of fasciolosis occurring in cattle-breeding farms where paramphistomosis is lacking and G. truncatula is absent.

A cornerstone in parasitology is why some species or populations are more parasitized than others. Here we examine the influence of host characteristics and habitat on parasite prevalence. We studied the helminths parasitizing the Pyrenean brook newt Calotriton asper (n= 167), paying special attention to the relationship between parasites and ecological factors such as habitat, sex, ontogeny, body size and age of the host. We detected two species of parasites, Megalobatrachonema terdentatum (Nematoda: Kathlaniidae) and Brachycoelium salamandrae (Trematoda: Brachycoeliidae), with a prevalence of 5.99% and 1.2%, respectively. Marginally significant differences were found in the prevalence between sexes, with females being more parasitized than males. The present results show significant differences in the body length of paedomorphic and metamorphic individuals, the former being smaller. Nevertheless, no significant correlations between parasite prevalence and either newt body length, ontogenetic stage or age were found. In comparison with other Salamandridae living in ponds, prevalence and diversity values were low. This may be due to a long hibernation period, the species' lotic habitat and its reophilous lifestyle, which probably do not allow for a high parasite load.

Enzyme-linked immunosorbent assays (ELISAs) which detect specific antibodies to fluke cysteine proteinases have provided good sensitivity and specificity for the immunodiagnosis of trematode diseases. To detect specific antibodies without the need for purified proteinase antigens, an immunocapture assay using Protein A was applied for the immunodiagnosis of paragonimiasis and fascioliasis. ELISA plate wells were coated with Protein A, incubated with diluted patient sera, then incubated with a preparation containing fluke cysteine proteinases, excretory–secretory (ES) products of adult Paragonimus westermani or Fasciola sp. The activity of fluke cysteine proteinases bound on the wells was measured by adding fluorogenic peptidyl substrate, Z-Phe-Arg-MCA or Boc-Val-Leu-Lys-MCA. This assay detected specific immunoglobulin G to cysteine proteinases of P. westermani and Fasciola sp. by measuring proteinase activity on the plate wells. Patient sera showed significant high values of proteinase activity when the wells were treated with the respective homologous ES products, whereas the sera had low values after treatment with the heterologous ES products. The sera of patients with other parasitoses and uninfected healthy individuals also showed low values after treatment with the above fluke ES products. Thus, Protein A immunocapture assay, which detected IgG specific for fluke cysteine proteinases, provided a high sensitivity and specificity for immunodiagnosis of paragonimiasis and fascioliasis.

Ten BALB/c mice were infected with the cestode Echinococcus granulosus. After the infection, serum was collected at different periods of time and monocyte chemotactic protein-1 (MCP-1) and macrophage inflammatory protein-2 (MIP-2) were determined. The level of MCP-1 increased from 290±12 pg ml-1 (mean±SD) at day 20 post infection (p.i.), to a maximum of 820±23 pg ml-1 on day 60 p.i., then decreased to 460±12.6 pg ml-1 on day 130 p.i. A second peak was observed at day 150 p.i. In addition, MIP-2 was detectable in serum as late as day 100 p.i. The highest level (100±11 pg ml-1) was observed on day 130 p.i., and decreased thereafter. Serum from noninfected animals (controls) contained no detectable levels of either MCP-1 or MIP-2. However, MCP-1 and MIP-2 appear to be implicated in E. granulosus infections, but their exact role during the disease is under determination.

Fasciola gigantica fatty acid binding protein (FABP) was evaluated for evoking an effective immune response in mice and rabbits, when delivered as a DNA vaccine in muscle cells. Polyethylenimine (PEI), 25 kDa, branched cationic polymer was used as a delivery vehicle for this DNA in the muscle cells of mice and rabbits. Naked DNA evoked mixed Th1 and Th2 responses in mice. PEI condensed DNA, at amine nitrogen over DNA phosphate (N/P) ratios of 4, 6 and 8 and with various DNA concentrations, failed to evoke a significantly higher antibody response compared to naked DNA in mice. Similarly, the humoral immune response to naked DNA administration in rabbit thigh muscles was poor and no boosting of this antibody response on administration of DNA complexed to PEI was observed. On metacercarial challenge, rabbits failed to show any significant protective immune response in both the naked DNA and PEI–DNA immunized groups. Administration of PEI alone (12.5 μg) in mouse thigh muscles caused significant muscle cytotoxicity but condensation of DNA with PEI had less of a toxic effect on muscle cells, which was inversely related to the N/P ratio. Delivery of plasmid DNA encoding F. gigantica FABP by high molecular weight polyethylenimine (branched, 25 kDa) did not boost the effective immune response in both the animal species, which could either be attributed to cytotoxicity associated with this cationic polymer or muscle cells being unsuitable target cells for PEI condensed DNA delivery.

Determinants of parasite species richness have been investigated in a host–parasite system comprising fish of the family Sparidae and their monogenean gill ectoparasites of the genus Lamellodiscus. This study was carried out on a small geographical scale in the northwestern Mediterranean Sea. Host phylogenetic relationships were taken into account by phylogenetic eigenvector regression which required the reconstruction of a phylogenetic tree for the sparid fish species using mtDNA sequences. Several ecological variables potentially acting on Lamellodiscus species richness were considered. Host body size and host migratory behaviour appeared to be the main determinants of parasite species richness in this system. It is concluded that structuring of monogenean communities is controlled more by ecological than evolutionary factors.

Rapid freezing and substitution with fixative prior to scanning electron microscopy was used to demonstrate the pattern of beat and recovery of the cilia of free swimming miracidia of Fasciola hepatica. There were stages of dexioplectic metachronal co-ordination and the power stroke was approximately 15° anticlockwise from the anterior–posterior axis. Around the circumference of the body of the miracidia there were approximately 12 metachronal waves of power and recovery. Free-swimming cercariae were recorded by time-lapse photography and, after conventional fixation, by scanning electron microscopy. Cercarial tail-beats were to the posterior of the body in the lateral plane at a rate of 8 Hz. The tail has paired lateral ridges positioned to act as leading edges. There is an array of 32 sensory papillae on the mid-ventral surface of the tail. The tegument of the most distal part of the tail is described: it is free of sensory endings and the surface shows a spiral pattern.

Galactosomum lacteum (Jägerskiöld, 1896) Looss, 1899 metacercariae, encysted on the optic nerve, on the brain and/or on the muscle and the connective of the pharynx and oesophagus, were found in Spicara maena L., S. flexuosa Rafinesque, 1810, S. smaris L. (Centracanthidae), Gobius cruentatus Gmelin, 1789 (Gobiidae), Symphodus tinca L., S. mediterraneus L. (Labridae), Serranus cabrilla L. (Serranidae), Diplodus sargus L. and D. annularis L. (Sparidae) caught in the Gulf of Cagliari (southern Sardinia, Italy). Excysted specimens were identified by some distinctive morphological features: more or less expanded forebody, depending on whether the specimens were living or fixed; tubular excretory bladder extending to the posterior border of the ovary; two-chambered seminal vesicle; asymmetrical and parenchymatous ventral sucker with lines of spines within its cavity; and unarmed gonotyle. Comparison has been made with the congeneric species metacercaria, G. timondavidi Pearson & Prévot, 1971, also registered in the Mediterranean Sea.

A further survey in East Caprivi, Chobe National Park, Okavango swamps and Kavango was undertaken in June 1976. No evidence of lechwe schistosomes was found in droppings of African buffalo (Syncerus caffer) nor baboons (Papio ursinus) living in lechwe habitats. It was thought that they were not capable of spreading or maintaining these parasites outside the confines of the known distribution of Kobus sp. The role of goats was equivocal but probably they too are poor hosts.

Kavango, an endemic area of S. haematobium and S. mansoni, was thought to be free of all animal schistosomes, thus confirming the hypotheses that (1) cattle and goats are poor hosts of the lechwe schistosomes and (2)S. mattheei was blocked from entering the territory by the presence of lechwe schistosomes in the surrounding areas. Evidence of schistosomes was not found in cattle and goats at Maun for the same reasons. The prevalence of S. mansoni at Maun has increased alarmingly over the past 20 years with a simultaneous disappearance of lechwe from the area. S. margrebowiei and S. leiperi eggs were found in lechwe and tsessebe droppings some 80 km north of Maun.

A high proportion of children with negative excreta from “non-endemic” areas in East Caprivi had positive CFT and/or skin tests, suggestive of exposure to lechwe schistosomes resulting in a possible immunity to S. mansoni and S. haematobium.

Researches involving dilution egg-counts, recently carried out by members of the staff of this Institute, have shown that the error variance, i.e. that between parallel counts, approximates more or less closely to the mean count. The following examples illustrate this point:—

The counts were all made by the same technique, and it is fair to total them in order to observe the average effect. This agreement between variance (or mean square) and mean is a feature of the Poisson distribution, and it has been assumed that dilution counts are in fact distributed according to the terms of that series, in all cases where the technique is adequate. Reasonable agreement between mean and error variance becomes, indeed, a criterion of the accuracy of the sampling technique. The purpose of the present note is to enquire a little further into the theoretical basis of this assumption, doubts having arisen as to whether high counts (say, up to 100) can be expected to conform to the Poisson series.

The essence of the McMaster (Gordon & Whitlock) technique is that one counts the eggs lying under a centimetre square engraved on a coverglass which is supported 0·15 cm. above a slide, i.e. the eggs contained in 0·15 cc. of faecal suspension. Since the suspending liquid is a half-saturated solution of salt, the eggs float up into one optical plane immediately below the coverglass, this making for ease in counting.

Since the publication of the writer's (1932) paper on the nematode genus Anguillulina, opportunities have occurred from time to time for making further observations on the morphology and anatomy of some of the species of the genus found associated with the roots of plants. Certain of them are parasitic in roots, at least during part of their life, at other times occurring free in the soil. Others have not been found within roots but have been obtained only in water extracts of turf and their exact relation to the roots of plants is at present undetermined.

Experiments were constructed to assess the ellects or changes in the diet of Periplaneta americana on Thelastoma attenuatum. Synthetic diets depressed parasitaemia although apparently adequate for the host. However, adult worms were supported best by high carbohydrate levels although protein was necessary for the final moult. Little effect was observed on the distribution of worms within the htndgut although excess roughage appeared to result in a slight backward migration. Egg output per worm remained fairly constant but rose slightly under conditions of extreme stress. Dietary changes affected the size of adult female worms differentially but had little apparent effect on food reserves. Possible reasons for the changes are discussed

1. Two new species, Spironoura sudanensis sp. nov. and Spironoura hexapapillata sp. nov. are described from freshwater fishes in the Sudan.

2. The genus Velariocephalus Singh, 1958 is considered to be a synonym of the genus Spironoura Leidy, 185G.

The daily cercarial output of two Nigerian strains of Schistosoma haematobium in sympatric Bulinus truncatus, B. globosus and B. senegalensis was measured at weekly intervals from the start of emission to the snails' death. In all cases cercariae were released throughout the life of the host, with no cases of “self cure”. Patterns of output through the course of infections in B. truncatus and B. senegalensis were similar to those reported for S. haematobium by other workers, with daily production of cercariae rising to a peak within a few weeks of the onset of shedding, then declining until the host's death. In the longer lived B. globosus production was significantly higher, but declined to very low levels after the initial peak; in some individuals cercarial output remained very low, while others showed a second period of high cercarial emission. The relative compatibility of each host-parasite combination is discussed.

The members of the genus Falcaustra are mainly parasites of tortoises. Of the 17 species of Falcaustra described from Asia, only 2 are parasitic in amphibians. Baylis (1933), recorded 3 species of Falcaustra from Malayan tortoises and Balasingam (1961), recorded another from Testudo emys from Malaya. The present paper records for the first time 3 new species of Falcaustra from Malayan amphibians for which the following names are proposed: Falcaustra purchoni sp.nov., F. pahangi sp.nov. and F. dubia sp.nov.