This work examines the ability of commerical zeolite Y to act as a slow release agent for a number of anthelmintic drugs. Administration to rats, dosed with Nippostrongylus brasiliensis, of pyrantel and/or fenbendazole and pigs, dosed with Ascaris and Oesophagostomum, of dichlorvos (DDVP) loaded onto zeolite Y was more sucessful in killing adult worms than administration of the pure drug alone. The zeolite Y was used as supplied for initial studies and then later dealuminated for further studies. The drug loadings were monitored by thermal analysis and the loaded zeolites were used in several field trials. The results indicate that zeolite Y is a suitable vehicle for the slow release of some anthelmintics. The slow release of drug from the zeolite matrix improved its efficacy.

The prevalence of helminths in pigs was investigated in five rural communities situated on the embankment of Dongting Lake in Zhiyang County, Hunan Province, People's Republic of China, in an area known to be endemic for Schistosoma japonicum. The helminth prevalences identified on the basis of faecal egg count analysis were: Oesophagostomum spp. (86.7%), Ascaris suum (36.7%), Metastrongylus spp. (25.8%), Strongyloides spp. (25.8%), Trichuris suis (15.8%), Globocephalus spp. (6.7%), Gnathostoma spp. (4.2%), Schistosoma japonicum (5.0%) and Fasciola spp. (1.3%). Post mortem examinations of a small number of pigs depositing eggs of different helminth species revealed the presence of Oesophagostomum dentatum, O. quadrispinulatum, A. suum, Metastrongylus apri, M. pudendotectus, T. suis, G. hispidum and Ascarops dentata. Prevalences of all helminths, with the exception of Oesophagostomum spp., were higher in young pigs (< 8 months old) compared with adult pigs. Prevalences of trematodes were very low, especially for S. japonicum which had decreased dramatically compared with previous reports from this area of P.R. China, whereas prevalences of nematodes were generally in agreement with those reported from other Yangtze River Provinces. Results from helminth prevalence studies in pigs, conducted in other provinces of P.R. China between 1987 and 1997, are presented and discussed. It was concluded that a government helminth control programme, implemented in 1995 to control S. japonicum infection in pigs in Hunan Province, may have resulted in a greatly reduced prevalence of S. japonicum in pigs in this region.

The effect of dose and duration of immunizing infections of Heligmosomoides polygyrus on protection against homologous challenge was studied in female TO mice. Primary infections were terminated at various levels with pyrantel embonate (adult infections) or ivermectin (larval infections) and mice were then challenged with 500 infective larvae (L3). The level of protection to secondary challenge positively correlated with the intensity of the primary immunizing infection but truncation of larval infection produced significantly better protection than termination of the adult nematode infection. The duration of the primary larval infection (1–6 days) positively correlated with the level of protection to secondary challenge, antibody responses and the proportion of circulating eosinophils. Histological changes in the gastrointestinal tract, peripheral leucocytic changes and antibody responses of the mice to H. polygyrus adult somatic antigens indicate both a cellular and humoral basis of host immunity to secondary challenge. Although the TO mice are slow responders in that they harbour chronic infections, immunization by intramucosal killing of the larval stage produced strong protection against secondary challenge infection. The presence of dead immunogenic larval stages within the intestinal wall may well be an important factor, since it exposes the host to stage specific antigens at an appropriate location. The implications of the findings for the control of gastrointestinal nematode infections are also discussed.

The lung fluke, Paragonimus westermani (Kerbert, 1878), is widely distributed in Asia, and exhibits much variation in its biological properties. Previous phylogenetic studies using DNA sequences have demonstrated that samples from north-east Asia form a tight group distinct from samples from south Asia (Philippines, Thailand, Malaysia). Among countries from the latter region, considerable molecular diversity was observed. This was investigated further using additional DNA sequences (partial mitochondrial cytochrome c oxidase subunit 1 (COI) and the second internal transcribed spacer of the nuclear ribosomal gene repeat (ITS2)) from additional samples of P. westermani. Phylogenies inferred from these again found three or four groups within P. westermani, depending on the method of analysis. Populations of P. westermani from north-east Asia use snail hosts of the family Pleuroceridae and differ in other biological properties from populations in south Asia (that use snail hosts of the family Thiaridae). It is considered that the populations we sampled can be divided into two species, one in north-east Asia and the other in south Asia.

Mature specimens of Cucullanus heterochrous Rudolphi, 1802 (Nematoda: Cucullanidae) were obtained from the intestine of the flounder, Platichthys flesus, from Danish waters. Eggs embryonate in seawater but do not hatch. Fully developed larvae pressed out of eggs are 430 μm long with amphids and dereids and enclosed within the cuticle of a previous larval stage. Infective larvae are believed to be in their third stage. Experimental studies showed that the polychaetes, Nereis spp., Scoloplos armiger, Brada villosa and Capitella sp., may act as intermediate hosts. In N. diversicolor the larvae increase their length to 1 mm within four weeks (15°C) without moulting. Experimental infections showed that larvated eggs are not infective to fish, whereas >550 μm long larvae from polychaetes survived in 4–24 cm long flounders and plaice, Pleuronectes platessa. Third-stage larvae 550 μm to 1.1 mm long were found in the submucosa of the intestine one week post infection. At a length of about 800 μm to 1.4 mm they moult to fourth-stage larvae. Fourth-stage larvae, immature and mature worms occur in the intestine and rectum. Fourth-stage larvae and adults survived experimental transfer from one flounder to another. Similar developmental stages survived for two weeks in the intestine of experimentally infected cod, Gadus morhua.

Soil samples from 79 sites on five islands of Indonesia were baited with insects for the recovery of entomopathogenic nematodes. Heterorhabditis and Steinernema were equally prevalent, and were recovered from 11.7% of samples representing 20.3% of sites sampled. Both genera were recovered from coastal sites only. Entomopathogenic nematodes were more prevalent on the Moluccan islands of Ambon and Seram than on Java or Bali. They were not detected on Sulawesi, where non-coastal sites only were sampled. RFLP analysis was used in the identification of nematode isolates. Heterorhabditis indica was the only heterorhabditid identified. Two RFLP types ofSteinernema were identified.

Samples of Lutjanus carponotatus(Lutjanidae) from reef flat (shallow) and reef slope (deep) sites around Heron and Wistari reefs on the southern Great Barrier Reef were examined for Pomphorhynchus heronensis(Acanthocephala). Individual fish from the reef slope had 0–9 (2.6) worms as compared with 1–122 (39.6) worms for individuals from the reef flat (P < 0.0001). Other variables (year, season, size of fish) made little contribution to the variation. Reef flat and reef slope sites were separated by as little as 300 m. These results imply both that the fish have very limited local movement and that transmission of the parasite is concentrated locally.

Investigations were undertaken to determine whether the tick Ornithodoros moubata moubata mounted a detectable immune response to primary and secondary infections with Acanthocheilonema viteae. Uninfected control tick survival rate was 70%, but only 45% in the primary infection group. Post-secondary infection survival rate (82%) was comparable to controls, indicating that these selected ticks had some protective advantage. Mean A. viteae infective larvae recovery from ticks with secondary infections was 31.4% lower than expected, suggesting the development of immunity. SDS–PAGE of haemolymph for proteins induced post-primary infection yielded a stronger signal at 45 kDa than controls, which was further elevated post-secondary infection. Proteins at 48, 22 and 16 to 18 kDa were detected in haemolymph from infected ticks but not seen from controls. The direct effect of haemolymph on microfilarial viability was examined using a novel in vitro assay; in these preliminary trials no differences were observed in parasite viability when exposed to haemolymph from infected or uninfected groups of ticks.

The ectoparasite fauna of two damselfishes, Stegastes nigricans and Dascyllus aruanus, from Moorea Island in French Polynesia was investigated. Gills of these damselfishes were infected with congeneric Monopisthocotylea Monogenea belonging to the genus Haliotrema. Stegastes nigricans were found to harbour a guild of three Haliotrema species whereas only one species inhabited D. aruanus. Microhabitat distribution, inter- and intraspecific competition and interspecific associations on the gill were studied. Observations on site preference revealed no spatial segregation between the three congeneric species inhabiting the gills of S. nigricans. Juvenile and adult monogeneans of that guild occurred on the same microhabitat. The dominant species Haliotrema sp. 1 did not expand on the microhabitat when the intensity of infection increased. Interspecific association tests revealed positive and negative associations. Haliotrema sp. 4 expanded its distribution on the gills of Dascyllus aruanus when the intensity of infection increased suggesting the likelihood of intraspecific competition. Juvenile and adult monogeneans of Haliotrema sp. 4 appeared to segregate as a result of intraspecific competition. This competition may exist to enhance resource availability when the gill habitat is limited. Overlaps between niche breadth and species microhabitat were revealed for monogenean species inhabiting S. nigricans. Interspecific competition did not appear to play an important role in the distribution of S. nigricans congeneric ectoparasites. Reinforcement of reproductive barriers may have led to the avoidance of hybridization.

The in vitro effects of ethanol and aqueous extracts of the medicinal plant Cardiospermum halicacabum on adult worms and microfilariae of Brugia pahangi were investigated. With or without the plant extracts in culture medium, the motility of adult worms, microfilariae and microfilarial release from female worms were monitored daily. After 7 days of culture, viability or tissue damage of adult worms was assessed using the MTT assay. At > 500 μg ml-1, the aqueous extract significantly reduced motility of adult females after 24 h of exposure and adult males after 3 days. The aqueous extract, at > 500 μg ml-1, also significantly reduced microfilarial release from female worms, starting on day 2. The reduction in the motility of adult worms and the pattern of microfilarial release from female worms were concentration and time dependent. The MTT assay results revealed that adult worms cultured in the presence of aqueous extracts at > 500 μg ml-1 were damaged. However, the aqueous extract did not affect the motility of microfilariae with the exception of those in higher concentration extracts. Higher concentrations of ethanol extracts (2 mg ml-1) inhibited both the motility of adult worms and the release of microfilariae from females. Little effect of ethanol extracts was detected by the MTT assay, as only slight damage was caused to worms exposed only to the highest concentration (2 mg ml-1). However, ethanol extract at 500 μg ml-1 rapidly reduced the motility of microfilariae on day 2. The present study revealed that an aqueous extract of C. halicacabum has mild but definite direct macrofilaricidal action on B. pahangi.

Five fish species used for preparation of a popular dish (cebiche) made with raw fish flesh in Mexico were obtained from five localities of the coast of Yucatan. Lutjanus synagris, Gerres cinereus, Sphyraena barracuda, Epinephelus morio and Haemulon plumieriwere examined for the presence of larvae of anisakid nematodes, causative agents of human anisakiasis. The nematode Pseudoterranova sp. was found in E. morio and S. barracuda with a total prevalence of 83% and 6.5 ± 6.2 worms per fish for E. morio, and a prevalence of 33% and 10.2 ± 30.0 worms per fish forS. barracuda. Contracaecumsp. was found to infect G. cinereus with a prevalence of 57% and 7.6 ± 11.4 worms per fish. The relatively high prevalence of Pseudoterranova sp. indicates that this parasite is a potential causal agent of anisakiasis on the coast of Yucatan. Although all larvae were found only in the mesentery of the fish host, their importance as a potential source of human infection cannot be excluded as larval migration to the muscles in dead fish is possible.

The spinous body tegument of the metacercaria of Timoniella imbutiforme (Molin, 1859) is described in detail and found to comprise an outer tegumental syncytium connected to subjacently situated subtegumentary ‘cells’. There are four types of secretory bodies in the outer syncytial layer as well as serrated overlapping spines and mitochondria. The subtegumentary ‘cells’ are characterized by the presence of four secretory body types as well as giant bodies which may be involved in the elaboration of the secretory bodies or spine material. The normal configuration of the somatic muscles of T. imbutiforme show that the muscular machinery necessary for activity once the larval stage becomes excysted is already in place. The sensory endings are found to be of the uniciliate type occurring in groups of up to eight in raised domes distributed over the body. Secretory gland cells are numerous and possess long ducts connected to the basal lamina of the outer syncytial layer via septate desmosomes.

An analysis is made of the community structure of the helminth parasites of the tuna Euthynnus affinis collected off the coast of Visakhapatnam, Bay of Bengal. The helminth fauna comprised 23 species, consisting of two monogeneans, 18 digeneans, one larval cestode, one nematode and one acanthocephalan. Didymozoids, represented by 14 species, were the dominant members of the parasite spectrum. The parasite assemblage is characterized by species richness as well as a high diversity. Most of the parasites found, the didymozoids in particular, were host specialists, exhibiting a high degree of host specificity. The fauna comprised three core species, four satellite species and many secondary species, with no potential for interaction among them but the degree of predictability is high in that each infracommunity comprised two or three core species superimposed by a few secondary species. Many host factors such as the varied diet, high vagility, long life span and endothermy appear to have contributed to the development of the species rich and diverse parasite communities in the tuna. There was evidence for a decrease in parasite density and an increase in diversity with increase in host size, indicating that host size has a profound influence on the component community structure.

Investigations were conducted on the host response in rainbow trout and the associated changes in mucous cell density during infection with the skin monogenean Gyrodactylus derjavini. Parasite populations increased on all naive hosts and peaked 4–5 weeks p.i. after which infection levels decreased. Introduction of naive fish into responding host populations resulted in heavy infections of the naive fish, whereas parasite expulsion continued in the responding host groups showing an acquired, non-sterile immunity. This non-sterile immunity lasted at least a month as these hosts were refractory to reinfection despite being exposed to a high infection pressure. Mucous cell hyperplasia was seen in some groups during the intermediary phase of infection, but at the termination of the study a significant depletion was evident. Passive immunization of naive host (with sera from immune hosts) did not confer protection. This indicates differences between host responses to G. derjavini compared to responses against other pathogens where such a passive immunity has been described.

The ultrastructure of the spinous body tegument of the metacercaria of Timoniella imbutiforme (Molin, 1859) has recently been described. Other regions of the metacercarial tegument, including those of the oral sucker, pharynx, and nephridiopore, demonstrate considerable specializations. The oral sucker tegument had an aspinous outer syncytial layer that possessed a pimpled apical surface as well as enclosing two types of secretory bodies. The pharyngeal tegument likewise lacked spines, but possessed only one type of secretory body, and a smooth but folded outer surface. The nephridiopore tegument, however, showed the greatest degree of specialization possessing a single type of secretory body specific only to this region of the tegument. Also associated with the syncytium here was a prominent long filamentous glycocalyx, and microtubules which were observed for the first time in this region of the tegument.

Experiments were carried out in vitro to investigate whether the sera of several animals as well as albumins and peptides might act as attractants for larvae of Strongyloides ratti. Samples of sera from several mammal species were dialysed and the aliquots were further centrifuged using ultrafiltration cartridges to remove any remaining small molecules. Additional test substances included commercially obtained ovalbumin, rat and bovine serum albumins, polypeptides such as peptone, tryptone and tryptose, amino nitrogens, monosaccharides, and reduced glutathione (triaminopeptide). Larvae were strongly attracted to the dialysed mammalian sera, which mainly consisted of serum albumin and globulins. Ov- and serum albumins, and polypeptides also acted as attractants. On the other hand, reduced glutathione, 16 kinds of amino acids and four kinds of monosaccharides did not attract this nematode.

A total of 110 cattle were examined in an area endemic for Bancroftian filariasis for the prevalence of infection of the bovine filarial parasite Setaria digitata. About 12.5% of cattle were found to harbour both adult worms in the peritoneum and microfilariae (mf) in circulation; 70% of the cattle were amicrofilaraemic but with an adult worm infection. A third group of cattle (16.5%) was free of detectable mf and adult worms. The presence of adult worms and/or mf did not influence the antibody levels to any of the four antigen preparations of S. digitata. However, there was a significant inverse relationship between the presence of antibodies to microfilarial sheaths and the absence of circulating mf as shown by the immunoperoxidase assay. Cattle immunoglobulin containing high titres of anti-sheath antibodies cleared circulating microfilariae very effectively in Mastomys coucha thus demonstrating the protective nature of anti-sheath antibodies in eliminating circulating microfilariae in vivo.

The infection by Opecoeloides furcatus and Poracanthium furcatum (Opecoeliidae) was studied in 121 Mullus barbatus and 113 M. surmuletus collected from the Spanish south-eastern Mediterranean. The prevalence of infection was most frequent in M. surmuletus with values of 81.42% for O. furcatus and 38.05% for P. furcatum. In M. barbatus the prevalences of O. furcatus and P. furcatum were 54.54% and 14.88% respectively. Statistically significant differences were found between the infection of the two hosts with P. furcatum. No significant differences in worm burdens could be attributable to host size or to seasonal changes, although a lower infection of M. barbatus by O. furcatus occurred in the autumn. Furthermore, the electrophoretic mobility of the enzyme malic dehydrogenase (MDH) was also studied and both digeneans presented different patterns, corresponding in both cases to homozygotic genotypes.

The ultrastructure of spermiogenesis and the mature spermatozoon in Catenotaenia pusilla (Cestoda: Catenotaeniidae) is described. Spermiogenesis is characterized by the presence of a single axoneme which grows on the outside of a cytoplasmic extension at an angle of 45°. Flagellar rotation and proximodistal fusion are produced in this process. The centrioles lack striated roots and an intercentriolar body. In the mature spermatozoon four different regions are described. The anterior extremity is capped by an apical cone and presents two helical crest-like bodies of unequal length. The axoneme, of the 9 + ‘1’ pattern of the Trepaxonemata, presents a periaxonemal sheath. The cortical microtubules form a spiral pattern at an angle of about 40° to the hypothetical spermatozoon axis. The nucleus is kidney- to horseshoe-shaped in cross section. Granules and proteinaceus walls are not observed in the spermatozoon of C. pusilla.

An in vitro assay was used to examine the effect of Bothriocephalus acheilognathi Yamaguti, 1934 (Cestoda: Pseudophyllidea) on the polarization response of pronephric leucocytes of carp, Cyprinus carpio. Leucocytes, isolated from naive, naturally-infected fish and carp injected intraperitoneally with cestode extracts, were exposed to parasite extracts (protein concentrations 0–10.0 μg ml-1), for up to 24 h in the presence or absence of carp serum. In general, polarization responses of the pronephric leucocytes, primarily neutrophils and eosinophils, increased with incubation time although there was no significant difference in the response induced by the different protein concentrations. Differences in the polarization response were, however, observed in naive, naturally infected and injected fish and the cells responded differently in the presence and absence of carp serum. In the absence of carp serum the polarization response of pronephric leucocytes in vitro was significantly reduced with cells obtained from injected and naturally infected fish compared with those obtained from naive carp. This suppression of leucocyte migration was however reduced by the addition of carp serum to the in vitro system. The role of this interaction between the possible suppression of polarization induced by the parasite and stimulation by serum is discussed.