The aim of this study was to develop a low-cost antifilarial immunoglobulin (Ig) G4 detection kit for the diagnosis of lymphatic filariasis. The kit was designed to be used by minimally trained personnel without the constraints of expensive laboratory equipment. We provide a description of the development and validation of a single-serum-dilution based enzyme-linked immunosorbent assay (ELISA) kit with ready-to-use reagents for measuring antifilarial IgG4 antibodies. The kit was tested on residents in Brugia malayi-endemic areas in southern Thailand. Detection was performed by naked-eye observation of the resultant colour of the immunological reactivity. The coefficient of variation (CV) was used to assess the reproducibility of the results. Long-term stability was measured over a 6-month period. Sensitivity of the test kit was 97% when compared with microfilariae detection in thick blood smears. Specificity was 98.7% based on the sera of 57 patients living outside the endemic areas who were infected with other parasites and 100 parasite-free subjects. All positive CVs were < 10%. The test kit was remarkably stable over 6 months. Field validation was performed by the detection of antifilarial IgG4 in 4365 serum samples collected from residents of brugian filariasis-endemic areas and compared with outcome colours of the test samples by the naked eye. Subsequent ELISA evaluation of these results using an ELISA reader indicated high agreement by the kappa statistic. These results demonstrate that the test kit is efficient and useful for public health laboratories as an alternative tool for the diagnosis of lymphatic filarial infection.

Experiments were carried out to obtain additional data concerning the role of IgM antibodies, specific for the cuticular surface of the microfilariae (mf) of A. viteae, in clearing microfilaraemia from high-and low-responder mice infected by transplanted adult worms. Although BALB/c mice, which sustain a chronic microfilaraemia, produced IgM mf surface-specific antibodies, the binding to target mf was weak when compared to that of antibodies from the serum of the resistant C57BL/10 mice. Furthermore, antibodies from BALB/c mice were not as efficient as those from C57BL/10 mice in promoting the adherence of immune or control leukocytes to mf in vitro. Evidence is provided to show that mf shed surface bound antibody. Although the results do not establish conclusively the mechanism underlying the contrasting response phenotypes of C57BL/10 and BALB/c mice, they provide support for the involvement of antibody in controlling microfilaraemia and suggest that quantitative and qualitative differences in the amount and affinity of IgM antibody specific for the mf surface, together with the natural tendency of the mf to shed surface bound antibody at 37°C. may combine to allow the former strain to clear microfilaraemia efficiently whilst the latter sustains a chronic infection.

The ultrastructure of the spermatid and spermatozoon of Macracanthorhynchus hirudinaceus (Archiacanthocephala) was studied by means of transmission electron microscopy. The flagellum and nucleus in the spermatid gradually expanded simultaneously. The karyoplasm of the spermatid transformed into dense inclusions and a multibarrel structure, which were also found in the spermatozoan body. The multibarrel structure was located close to the flagellum and consisted of many irregular microtubes. The flagellum of the developing spermatozoon was observed in a concavity of the spermatid nucleus. The microtubule arrangement of the flagellum was ″9+2″. No mitochondria or acrosome were observed in spermatozoa.

Stunkardia minuta sp. n. was recovered from the small intestine and rectum of 5 box-tortoises (Cuora amboinensis) in Malaysia. The average body size (L × W) is 11·42 × 2·35 mm; oral sucker 1·51 × 1.02; oral pouches 0·21 × 0·18; ventral sucker 1·67 × 1·43; oesophageal bulb 0·54 × 0·54; ant. testis 0·95 × 0·98; post. testis 0·94 × 0.94; seminal vesicle 0·82 × 0·24; ootype 0·21 × 0·41; ovary 0·41 × 0·34; and egg 121 × 83 μm. Although morphologically similar to S. dilymphosa, S. minuta is distinct from any other reported member of the Paramphistomidae.

An attempt was made to identify the human hookworm involved in failed-treatment cases using abnormal hosts and scanning electron microscopy. Thirty-seven, 2 to 6 month old Chinese hamsters (Cricetulus griseus) from a closed, outbred, conventional colony, were each given between 20 and 120 filariform larvae per os. The larvae were cultured from faeces from mebendazole (Vermox®) 500 mg single-dose, failed-treatment cases living in the lowveld farming area of the Transvaal Province, South Africa. About 60 to 78 days after inoculation, the animals were killed and adult worms were removed from their small intestines. Eleven (30%) of the 37 hamsters harboured a total of 31 adult worms (19 males and 12 females), while 26 hamsters were refractory to infection. The greatest number of worms recovered from a single animal was six. A total of 27 worms (17 males and 10 females) were subjected to examination by scanning electron microscopy. Micrographs showed male and female worms to be morphologically all of the Necator americanus species, as identified by a pair of ventral and dorsal cutting plates, a dorsal tooth and the fused terminus of spicules in the male bursa. The transverse cuticular striations were distinct and smooth. Several points of interest arose from the results of this study and are discussed.

The initial pathway of skin penetrating larvae of Strongyloides ratti inside the host is not systemic and could well involve local components of the lymphatic system. The experiments described were an attempt to detect an effect on immunity depending on whether larvae of a challenge infection were committed to a pathway through heavily primed or lightly primed lymph nodes. Female rats were immunised by subcutaneous injection into the right forearm of 1000 heat killed, or 1000 live, third stage larvae of S. ratti. Animals given living parasites were placed on a diet containing 0·1 or 0·2% thiabendazole 48h or 36h after ifection. Challenge infections of <100 larvae (‘exact’ doses) were applied on day 21 to either the right or left flank and, similarly, to controls that had received the anthelmintic but not the priming infection. Heat killed parasites elicited no response at all. Priming with live larvae stimulated a significant immunity (16% and 37% depression in 2 experiments), but there was no difference between rats whose challenge was on the same side as the priming dose and those which had the contralateral treatment. The significance of these results to theories of pathfinding in general, and to practical immunology, is discussed.

Promising results in the colorimetric assay of potato-root eelworm hatching factor have been obtained with picric acid and with 3: 5- dinitrobenzoic acid. Both of the reagents are used for the assay of cardiac glycosides with which the hatching factor may have affinities.

Thirty-eight spotted ground squirrels, Spermophilus spilosoma were examined for endoparasites, and 31 of them were infected with one or more species. Helminths found were Hymenolepis citelli, Citellina triradiata, Physaloptera massino, and Trichuris sp. The protozoan Eimeria larimerensis was also found. Citellina triradiata is redescribed. This is the first report of any helminths from this squirrel.

As a result of complaints received from farmers as to the presence of parasitic worms in sheep, an attempt was made during the period October 1923 to September 1924 (inclusive) to ascertain the distribution and relative abundance of the more usual parasitic worms inhabiting the alimentary canal of sheep in North Wales. Regular collections were made from the following centres:—Pwllheli, Carnarvon, Bangor, Conway and Llandudno, with a few records also from Colwyn Bay, Bethesda, Menai Bridge and Denbigh. All the sheep examined were slaughtered for food purposes, practically the whole being obtained from slaughter houses. Almost all had grazed for considerable periods on the farms in the district mentioned, at elevations varying from sea level to 600 feet.

On the 17th September, 1964, a Mute Swan, Cygnus olor (Gmelin) was received for examination. It was a male bird, estimated to be about three years old, which had been found at Henley-on-Thames, Oxfordshire, and had been killed on account of a broken wing which had healed badly. It was believed to be of wild stock.

A detailed examination of the abundant flatfish species Platichthys flesus, the flounder, in the tidal Thames has revealed the presence of four digenean metacercarial parasites, Cryptocotyle concava (Creplin, 1825), Timoniella imbutiforme (Molin, 1859), T. praeterita (Looss, 1901) and Labratrema minimus (Stossich, 1887). Flounders were recorded as a new second intermediate host for T. praeterita and L. minimus. They were also recorded as second intermediate hosts for the first time in British waters for T. imbutiforme. The temporal and spatial characteristics of these infections were examined and were believed to provide indirect parasitological evidence of the movement patterns of flounders during their utilization of the Thames Estuary as a nursery ground. From these data it was also surmised that the first intermediate host of T. imbutiforme, T. praeterita and C. concava was probably the molluscan species Hydrobia ulvae in the lower Thames Estuary, whereas L. minimus was most likely to occur in the molluscan host Cerastoderma edule, also present in the lower estuary.

Genetic markers, DNA sequences and karyotypes, of some European lissorchiid species from their intermediate and final hosts were obtained to clarify controversial data about their life cycles and taxonomy, and to reveal phylogenetic affinities. The life cycles of three species have been confirmed for the first time based on molecular data. Comparative analysis of internal transcribed spacer 2 (ITS2) and partial 28S rDNA sequences has undoubtedly proven that cercariaeum of type-species of the genus Asymphylodora, Asymphylodora tincae, develops in pulmonate snails, Anisus vortex and Stagnicola palustris, but not in the genus Bithynia. The faucet snail, Bithynia tentaculata, serves as the first intermediate host for Parasymphylodora (=Asymphylodora) markewitschi and Parasymphylodora parasquamosa; adults of both species were isolated from the common rudd, Scardinius erythrophthalmus. It has also been confirmed that B. tentaculata serves as the second intermediate host for P. parasquamosa. Phylogenetic analysis supports the validity of the genus Parasymphylodora. Two species, Parasymphylodora markewitschi and P. parasquamosa, with cercariaeum belonging to the Parasquamosum group, are closely related and are being recovered as a well-defined evolutionary lineage in phylogenetic trees. A significant divergence between Parasymphylodora spp. and Asymphylodora spp. was revealed. The diploid chromosome set of P. markewitschi is composed of 14 chromosomes and does not show similarities with karyotypes of other lissorchiid species. Asymphylodora progenetica and Asymphylodora tincae share the basal diploid value of the family, 2n = 20, and reveal very close morphology of the corresponding chromosome pairs. Karyotypic similarities of these species are in accordance with molecular phylogenetic data. Thus, the available molecular and cytogenetic data support the assignment of P. markewitschi and P. parasquamosa to a separate genus, meanwhile, the assignment of A. progenetica to the genus Parasymphylodora was not justified.

Brachylaemus dasyuri (Johnston, 1912) is recorded from Thylacis obesulus (Shaw & Nodder, 1797) from Queensland. It is redescribed and Brachylaemus simile (Johnston, 1912) is confirmed to be a synonym.

A review is made of the species of Brachylaemus recorded from American and Australian marsupials. It seems probable that they have arisen from a common ancestral type parasite in an ancestral marsupial.

Early in June, 1933, the writer received two vials of caterpillars, to which nematodes were attached, from Mr. A. M. Adamson of the Pacific Entomological Survey with a request, supported by Mr. E. P. Mumford, the Director of the Survey, that an examination and identification of the helminths would be much appreciated.

A gelatin-agar bolus, designed and developed for the administration of metacercariae of the liver fluke Fasciola hepatica, was evaluated in adult Holstein Friesian cattle. The metacercariae, enclosed within a gelatin capsule, were placed inside the bolus and delivered to each animal using an oesophageal balling gun. At slaughter, 13 weeks after challenge, an average of 25% of the challenge dose was recovered from each liver. This percentage recovery is similar to that obtained with other known methods. The new bolus, however, offers improved handling qualities. In addition, the bolus also has potential for improving a number of other techniques including those for the administration of other parasites, compounds or chemotherapeutic agents.

A successful peritoneal transplant of 5 Cardiofilaria nilesi adult worms, 3♀ Symbo; and 2♂ Symbo; is reported. Microfilaraemia in the recipient chicken was found on the 6th day after the transplant. The count rose to 4/20 c.mm., from 1/20 c.mm. in 19 days after the transplant. The highest count recorded was 14/20 c.mm. The microfilaraemia persisted for 63 days.

Neurocysticercosis is a parasitic disease caused by encysted larvae of Taenia solium in the human central nervous system. Cysts mainly affect the cerebral hemispheres, although they can also be found in ventricles, basal cisterns, and subarachnoid spaces, and rarely in the cerebellum. Given the impossibility of studying the disease in human patients, Cardona et al. (1999) developed a mouse model of neurocysticercosis, using Mesocestoides corti, a closely related cestode. This allows us to study the parasite–host relationship and the mechanisms involved in the disease, in order to improve the therapy. In this murine model of neurocysticercosis, the location of tetrathyridia in parenchyma, ventricles and meninges has already been reported. The aim of this work is to report the cerebellum as a new location for M. corti tetrathyridia in the murine model of neurocysticercosis. A murine model that reproduces the human pathology is essential to evaluate the symptomatology and response to drug treatment in experimentally infected mice.

In this study, morphological and molecular features were used to identify a new Steinernema sp. from Chhattisgarh, India. Morphological and molecular features provide evidence for placing the new species into the “bicornutum” clade. The new species is characterized by the following morphological features: infective juveniles with a body length of 587 (494–671) μm; a distance from the anterior end to excretory pore of 46 (43–50) μm; a distance from anterior end to nerve ring of 72 μm (61–85 μm); and E% of 88 (77–97). The first-generation males are characterised by 27 genital papillae and very short spicules, with a length of 61 μm (53–67) μm. The SW% and GS% ratio of S. shori n. sp. are 139 (107–190) and 75 (62–90), respectively. The new species is further characterized by sequences of the internal transcribed spacer and partial 28S regions of the ribosomal DNA. Phylogenetic analyses show that S. shori n. sp. is most closely related to S. abbasi, S. kandii, and S. yirgalemense.

The present report describes two cases of infection by Molossinema wimsatti in the brain of Pallas's mastiff bats (Molossus molossus). The first bat was captured and killed by a domestic cat in a suburban area of the municipality of Patos, Paraiba, northeastern Brazil. The second bat was found crawling on the ground in the same area before dying. No gross lesions were found at necropsy. Histology of the central nervous system revealed filarioid nematodes in the brain ventricles and cerebellum. There were adults, subadults and eggs, the latter sometimes containing microfilariae. No inflammatory response was observed in bat 1, while bat 2 presented a mild lymphoplasmacytic meningoencephalitis. Three nematodes were recovered and submitted for parasitological examination. The diagnosis of M. wimsatti infection was based on the histomorphological and parasitological characteristics of the agent and its location in the brain ventricular system of insectivorous bats. The infection likely occurs in other insectivorous bats from South American and Caribbean countries but may be overlooked.