Epidemiological studies of Echinococcus multilocularis infections in definitive hosts require a reliable and economic diagnostic method. In this study, the current copro-DNA examination technique was modified by increasing the faecal amounts tested and adding a step to neutralize the faeces before DNA extraction. Reliability of the modified method was evaluated using rectal faecal samples from red foxes and comparing them with intestinal worms detected using the sedimentation and counting technique (SCT) following necropsy. The modified copro-DNA examination method demonstrated 93.9% sensitivity (138/147) on the SCT. Its detectability increased depending on the worm burden, and the sensitivity was 100% in cases harbouring over 1000 worms. From 111 SCT-negative cases, six (5.4%) were copro-DNA-positive, and all were confirmed as E. multilocularis via sequencing analysis. Five of the remaining 105 SCT-negative cases (4.8%) retained polymerase chain reaction (PCR) inhibitors in the extracted solution, suggesting that approximately 5% of the red fox faeces retained these inhibitors after treatment with the present copro-DNA extraction method. Although further evaluation is needed for faeces deposited in the wild, the present copro-DNA examination technique will help monitor the E. multilocularis prevalence in definitive hosts. When used for detailed evaluations of endemicity (e.g. changes in infection pressure or spread in non-endemic areas), the absence of PCR inhibitors should be confirmed, and multiple trials on faecal subsamples are recommended.

Comparative histochemical studies of glycosidase activity were carried out in Clonorchis sinensis, Eurytrema pancreaticum, Fasciola hepatica, Dipylidium caninum, Hymenolepis nana, Ascaris suum, Toxocara canis, Ancylostoma caninum, Trichuris vulpis and Dirofilaria immitis. The enzymes examined were: N-Acetyl-α-Glucosaminidase (Ec 3.2.1.30), α-Glucuronidase (Ec 3.2.1.31) and α-Galactosidase (Ec 3.2.1.23). There were variations in enzyme distribution and intensity among the species and also between trematodes and nematodes; no marked positive reaction for these enzymes occurred in cestodes. In some trematodes, the caeca, especially in the brush border, and the tegument, subtegumental cells and testes, were reactive to the enzymes. In nematodes, although there was variation in reactions among species, N-acetyl-α-glucosaminidase and α-galactosidase were localized in the hypodermis and lateral cords excluding the excretory canal, and coelomocytes, intestinal epithelium and the walls of the reproductive systems.

1. Nematobothrium labeonis n. sp. from the orbit of three species of Labeo is described.

2. The more recent literature relating to the classification of the genus is considered.

3. N. labeonis is contrasted with related species.

In 1988, 1989 and 1990 second year grazing heifers, naturally infected with gastrointestinal nematodes, were separated into two groups, one of which was treated orally with albendazole. In 1988 and 1989 treatment was administered immediately after parturition (February), and in 1990 during the last term of pregnancy (December). Both treated and control animals were grazed on separate plots in a rotational system. Maximum faecal egg counts were observed amund parturition, except in 1990, when treatment was given at the end of gestation. The main genera identified were Cooperia, Trichostrongylus, Ostertagia and Oesophagostomum. The number of Ostertagia larvae in the treated groups increased from 1989 to 1990, while the others decreased. Pasture contamination with third stage larvae (L3) was lower on the piots grazed by treated heifers. Maximum numbers of L3 were found in autumn, at the end of winter, and at the beginning of spring. Mean serum pepsinogen concentrations were significantly higher in the untreated groups. This concurs with the pattern for L3 on pasture. The trial shows that if a single treatment against gastrointestinal nematodes is carried out, and the animals remain on contaminated pastures, the parasitic load tends to level out after 4–5 months under favourable climatic conditions. However, the percentages of nematode genera occurring in the new populations may differ from those in the original infection.

In connection with investigations of the United States Bureau of Fisheries in Alaska in 1906, Professor H. B. Ward spent considerable time in the study of the parasites of the Pacific salmon. Around Excursion Inlet he secured a large amount of valuable material. The following study was made of one of the specimens collected by him, to whom I am grateful for the loan of it.

It has been, for some time, a recognised fact that intense eelworm infections are, in some instances, accompanied by only slight damage to the plants, while in other areas where the infection is much less intense, the plants show every symptom of “potato sickness” and the crop is a complete failure.

ALTHOUGH some marsupials are insectivorous or carnivorous in diet many others are vegetarian in diet and frequently cause appreciable damage to pastures and crops and therefore are of considerable economic importance. Recently, during the course of searching the literature for references to helminth infections in marsupials it was observed that, in certain cases, hosts other than marsupials were involved and in some instances were of economic importance in that they were domesticated animals. Whether or not marsupials assist in the spread of parasitic infection is somewhat inadequately dealt with in the literature but one cannot lose sight of their acting as reservoir hosts.

1. Four worms were recovered from the subcutaneous connective tissue of a leopard. Of these two males and one female were identified as Filaria martis Gmelin, 1790 with the measurements given and a female described as a new species under the name Filaria russelli.

2. This finding is the first record of the genus Filaria Mueller, 1787 in a leopard.

A medium containing inorganic salts, vitamins and amino-acids is described for the in vitro cultivation of the amphistome, Orthocoelium scoliocoelium, from the rumen of buffalo. Based on the properties of rumen fluid, this medium promoted and sustained normal egg production, at 37°C temperature and pH 7·4 to 7·8 for 15 days. In contrast to other media used, no patho-physiological alterations except abnormal cytoplasmic lipid, were detected. Since the parasites produced normal eggs, it is concluded that this medium is suitable for short-term cultivation of the parasites.

Infective larvae (third-stage larvae) of both Angiostrongylus cantonensis and A. costaricensis from their snail intermediate host were subjected to either X-ray or gamma-ray irradiation. The viability of the irradiated larvae was assayed by oral inoculation of the larvae into rodents (A. cantoflensis in mice and rats, A. costaricensis in mice only). From the results of worm recovery, the minimal dose of irradiation that inhibited the infectivity of the third-stage larvae of A. cantonensis and A. costaricensis was 2 and 4 kGy, respectively.

Two new species belonging to two different families of the super-family Aphelenchoidea are described. The material was obtained from the roots of tobacco plants, Nicotiana tabacum.

The worms were killed by hot water, fixed in F.A.A. solution, cleared in 1% glycerine alcohol mixture and mounted in pure glycerine.

Three new records are made, viz: Achillurbainia recondita Travassos, 1942; Metadelphis evandroi Travassos, 1944; Rhopalias coronatus (Rudolphi, 1819) Stiles and Hassall, 1898; all are recorded from the manicou, Didelphis marsupialis insularis (Allen) from Trinidad, British West Indies. Descriptions of all these species are given.

This study reports further observations on encystment and excystment of the paramphistomid trematode Zygocotyle lunata. Of numerous substrates tested in the laboratory for cercarial encystment, i.e. plastic and glass dishes, Styrofoam™, aluminium foil, snail shells, and cheesecloth, all but cheesecloth allowed for 100% encystment within 4 h of cercarial release at 28°C. Numerous physical and chemical factors were tested to kill cysts, and their capacity to do so was determined by the subsequent inability of cysts to excyst within a chemical excystment medium. Vinegar, Italian salad dressing, soy sauce and 10% solutions of sucrose, acetic acid and sodium chloride were the most detrimental. Freezing, boiling and storage of cysts for more than 2 years also killed them. Several marinades such as lemon juice, Worcestershire sauce, and various concentrations of potassium permanganate, did not kill the majority of cysts tested. Since Z. lunata is a potential foodborne trematode for humans and animals, it is important to determine suitable ways to control or kill these cysts.

A defined medium is described as a basis for in vitro culture work with larval Fasciola hepatica. This medium, termed BCM, can be quickly made up by using a system of stock solutions. BCM contains inorganic salts, glucose, amino acids, vitamins and antibiotics, but no lipid or proteins. Rediae can be dissected from infected snails for culture, but many appear to be contaminated with bacteria. Large rediae cannot survive in BCM but free immature cercariae can complete their final maturation in vitro. This final maturation, from the 30th to the 35th day after miracidial penetration of donor snails, includes tail growth and appearance of body pigmentation. Cercariae matured in vitro encyst successfully when transferred from BCM to water. Small rediae survive in BCM for 5 days, but show no growth or development measured as dry weight and total nitrogen.

The lungworm, Aelurostrongylus abstrusus, is a common parasite of the domestic cat in Great Britain and produces pulmonary lesions in that species which are occasionally severe. The disease may be reproduced experimentally either by the oral administration of third stage larvae which have been extracted from infected molluscs or by feeding the latter, themselves. For experimental purposes, the former procedure is preferred but the main disadvantage is that an infecting dose must be given on several occasions and over a period of days if sickness, with consequent loss of larvae, is to be avoided. Parenteral administration would, therefore, be advantageous. It is known that pulmonary infections have been produced in calves by the subcutaneous administration of larvae of Dictyocaulus viviparus (Wade and Swanson, 1958), in pigs by the intravenous and intratracheal injection of Metastrongylus spp. larvae (Kelley & Krous, 1961) and in sheep by the intravenous inoculation of Dictyocaulus filaria larvae (Michel and Sinclair, 1963). As neither intravenous nor intratracheal administration is particularly desirable in cats, it was resolved to attempt the reproduction of lungworm disease in that species by the subcutaneous injection of third stage larvae of Aelurostrongylus abstrusus.

The relationship between the cercarial allergen and two previously isolated egg allergens (J1, J2) of Schistosoma japonicum was examined especially in terms of the cross-reactivity between them. Semi-purified cercarial allergen (JAC) was obtained from the crude extract of S. japonicum cercariae by gel chromatography on Sephadex G-200. The apparent molecular weight of JAC was estimated approximately as 60–100 kDa. JAC could bind to Con A-Sepharose, indicating its glycoprotein nature. Three groups of BALB/c mice were immunized with JAC, J1 or J2 using A1(OH)3 as adjuvant, and the cross-reactivity of each antiserum was examined by PCA. Anti-JAC, anti-Jl or anti-J2 serum was highly specific to the corresponding antigen. When IgE-ELISA of S. japonicum patient sera was performed using JAC, J1 or J2 as an antigen, the correlation between anti-J1 and anti-J2 (r = 0.78) was high, whereas the correlation between anti-JAC and anti-J1 (r = 0.27) or between anti-JAC and anti-J2 (r = 0.12) was low. These results suggest that most IgE epitopes on cercarial allergen are independent from those on egg allergens in S. japonicum.

In 1923, the writer in studying some species of the family Trichostrongylidæ, had occasion to create a new genus Graphidioides to receive the form previously known as Graphidium affine (Megnin, 1895), Railliet and Henry, 1909. This species differs from the type species of the genus Graphidium on the following points: absence of cervical papillæ, shape of bursa and disposition of ventro-ventral rays, shape of accessory piece and spicules, shape of the ovejectors and vagina and posterior extremity of the female. In an addendum on the remaining species of Graphidium, G. rudicaudatum the following statement was made:—

“Graphidium rudicaudatus R. & H., 1909, from Viscacia viscacia from Argentina resembles G. affinis very closely. It differs mainly by its smaller size, equal ovejectors, sharply pointed externo-dorsal ray and absence of an accessory piece. Its affinities to Graphidioides are so close, that the latter point requires re-investigation. The other points are only of specific value, and it is proposed to include it in this new genus. Should the absence of an accessory piece be confirmed, its position would require re-determination.”

The parasite was found in the bile ducts of three sperm whales, which were caught in the 1957–58 whaling season.

As specimens were found independently in bile ducts of three whales this position in the host appears to be the natural habitat. All the worms were in pieces and incomplete. The pieces, in each case, totalled approximately 18 metres, 0·9 metres and 21 metres respectively. The holotype (length 18 metres) was probably complete except for a few terminal proglottids because it consisted of two very long pieces and one short piece which could be fitted together. The type material has been deposited in the British Museum (Natural History) Registered No. 1961·12·12·1 (Holotype) and 1961·12·12·1 (Para types).

Strongylus (Dicerocola) tremletti sp.nov. is described from the black rhinoceros, Diceros bicornis L. from Kenya.

The affinities of S. (D.) tremletti sp.nov. with the other species of the genus Strongylus Müller, 1780 are discussed and the subgenus Dicerocola subgen.nov. is erected to contain it.

The species of Strongylus in Kenya are noted and original drawings of S. (D.) asini Boulenger, 1920 are included.