A clone, designated as TsTM, was selected from the cDNA library of newborn larvae (NBL) of Trichinella spiralis through immunoscreening against infected sera. The clone contained a cDNA transcript of 855 bp in length with a single open reading frame, which encoded 285-amino acids (33 kDa in the estimated molecular weight). A sequence analysis revealed that the clone TsTM encoded the full-length of tropomyosin gene. The phylogenetic analysis of the tropomyosin gene was in good agreement with the classical taxonomical position of T. spiralis. The fusion proteins encoded by the clone TsTM were produced in an Escherichia coli expression system and affinity purified, and the antibody was raised against the protein for the following studies. The antibody against the fusion protein positively bound to the hypodermal muscle layer in immunolocalization analysis, and the 35 kDa band in crude extracts of muscle larvae but not in excretory and secretory (ES) products on Western blots. The antigenicity of the clone TsTM was recognized by host mice but exhibited little species specificity.

Isoelectric focusing of glucose-6-phosphate dehydrogenase (G6PD) produced clearly identifiable profiles for S. haematobium and S. intercalatum and their hybrids. To provide a more detailed analysis of the interactions of S. haematobium and S. intercalatum in South West Cameroon over the last 12 years, G6PD analyses were carried out on individual schistosomes collected in Kumba in 1990, Loum in 1990, 1999 and 2000 and Barombi Mbo and Barombi Kotto in 1999. Studies were also carried out on the two parental species S. haematobium Barombi Mbo, S. intercalatum Edea and subsequent generations of hybrids resulting from laboratory crosses of the two parental species. The isoenzyme analysis demonstrated that the 1990 isolate from Kumba, was a recombinant of S. intercalatumxS. haematobium, and that 30% of individual schistosomes collected in 1990 in Loum were also recombinants. The remainder gave data indicative of S. haematobium. In 1999, 12.5% of individuals from Loum showed recombination and 10% in 2000. Results from the most recent parasitological survey in October 2000 showed the persistence of the recombinant population in addition to that of S. haematobium. There was also evidence of recombination having taken place in Barombi Kotto but not Barombi Mbo. This study demonstrates how the situation has changed over the last 12 years, and emphasizes the importance of assessing morphological, biological and molecular data together to gain a true picture of the rapidly evolving situation.

Little is known about immune responses in teleosts as linked to the aetiology of pollutants and parasitic diseases and in particular their combined effects on the host. Cadmium(Cd)-mediated immunological responses in the thymus and pronephros of juvenile carp (Cyprinus carpio), experimentally infected with the blood parasite, Sanguinicola inermis (Trematoda: Sanguinicolidae) for 30 days followed by an exposure to 0.1mg Cd2+ l−1 for 48 or 168h were investigated. Differential organ-specific changes occurred in both organs examined. In carp exposed to Cd, intracelluar spaces, vacuolation in the eosinophils, dissociation of cell membranes together with the formation of concentric whorls occurred. The thymus of infected carp exposed to Cd had a granular cytosol which contained vesicles with electron-dense inclusions, swollen mitochondria with distended cristae and condensed nuclei in the erythrocytes. Cell counts on the two organs revealed a differential response to cadmium exposure in S. inermis infected carp compared to control infected fish. A significant increase in the neutrophil, eosinophil and thrombocyte components occurred in the thymus in contrast to a significant decrease in pronephric neutrophils. In addition, there was a differential blastogenesis response in infected and Cd-exposed infected carp fry exposed to cercarial antigens and the mitogens, concanavalin A and pokeweed mitogen.

Adult Ascaris suum were dissected to obtain different worm components (body wall, body fluid, ovaries, uterus and oesophagus) which were used as antigens when testing 95 sera of naturally A. suum-infected Iberian pigs by enzyme-linked immunosorbent assay (ELISA) and Western blot (WB). Pigs with patent Ascaris infections had significantly lower ELISA optical density values than pigs without adult worms when using the body fluid and the body wall as antigens. A poor negative correlation was found between adult intestinal worm burden or eggs in faeces and specific antibody responses, measured by ELISA and WB using all antigens. By WB, the recognition of specific bands was variable, but three groups of bands with molecular weights of 97 kDa, 54–58 kDa and 42–44 kDa were generally recognized by sera from naturally infected pigs as well as from hyperimmunized pigs when using the five antigen extracts. The ELISA and WB techniques may be used for immunodiagnosis, using somatic adult worm antigens, to declare young pigs to be Ascaris-free but cannot be used for individual Ascaris-diagnosis in adult Iberian pigs.

Laboratory investigations on three aquatic populations of Galba truncatula, originating from the Peruvian Altiplano and French Massif Central, were carried out during three successive snail generations to determine if these populations might be successfully used for the metacercarial production of Fasciola hepatica under experimental conditions. High numbers of surviving snails at day 30 post-exposure (>70%), high prevalences of F. hepatica infections (>60%), and prolonged productions of cercariae for a mean period of 35 to 47 days were observed in the three populations, whatever the snail generation. In the Peruvian population, metacercariae of F. hepatica significantly decreased in numbers from a mean of 251 in the parent snails to 124 per snail in the F2 generation, whereas no significant variation was observed in the two French populations. As these aquatic snails rarely emerged out of water, the use of these populations for the commercial production of F. hepatica metacercariae was of great interest, because the daily time spent watching the breeding boxes of snails was clearly shorter, thereby reducing the cost of producing metacercariae compared with using amphibious snails reared with romaine lettuce.

Dexamethasone, a known immunosuppressant, was administered by bath or injection to Atlantic salmon Salmo salar (Conon stock) to study if this treatment could affect the susceptibility of fish to infections with a Danish strain of Gyrodactylus derjavini (Monogenea). Three groups of S. salar (Conon stock) were immersion treated either with 10, 60 or 240 μg dexamethasone l-1 water, respectively. In addition, one group (positive control) was treated intraperitoneally with 200 μg dexamethasone per fish and one negative control group was kept untreated. A single G. derjavini parasite was placed on the anal fin of each fish and the infection was subsequently monitored weekly for 6 weeks. An increase in parasite populations on the salmon was positively correlated with the amount of immunosuppressant used. Infection levels in the group immersion treated with dexamethasone (240 μg l-1 water) and in the i.p. treated positive control group were significantly higher compared to the untreated control group.

To determine the role of interleukin-5 (IL-5) and eosinophils in protection against Strongyloides ratti, mice treated with anti-IL-5 monoclonal antibody (mAb) were infected with S. ratti larvae. Strongyloides ratti egg numbers in faeces (EPG) in mAb treated mice were higher than those in control mice on days 6 and 7 after inoculation. The numbers of migrating worms in mAb treated mice 36h after inoculation were higher than those observed in control mice. Intestinal worm numbers in mAb treated mice 5 days after inoculation were higher than those in control mice. These results show that eosinophils effectively protected the host against S. ratti infection by mainly the larval stage in primary infections. The involvement of eosinophils in protection against secondary infection was also examined. Before secondary infection, mice were treated with anti-IL-5 mAb and infected with S. ratti. Patent infections were not observed in either mAb treated or control Ab treated mice. The numbers of migrating worms in the head and lungs of mAb treated mice increased to 60% of that in primary infected mice. Intestinal worms were not found in mAb treated mice or in contcrol mice after oral implantation of adult worms. Eosinophils were therefore mainly involved in protection against tissue migrating worms in secondary infections.

The effect of pollutants on the intensity of infection of metazoan parasites in the Mayan catfish, Ariopsis assimilis was investigated. Data were collected on pollutants and metazoan parasites from 76 catfish from five localities in Chetumal Bay in October, 1996. Nineteen pollutants (pesticides, polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs)) were found in the catfish livers. Heavy metal content was not determined. Nineteen metazoan parasite species were recovered. After controlling for fish length and sampling station, there was a significant negative linear relationship between the intensity of the larval digenean Mesostephanus appendiculatoides and 1,1,1,-trichloro-2,2-bis (4-chlorophenyl) ethane (DDT) concentrations. This negative relationship may be explained either by the effect of the pesticide on the mortality of (i) free-living larval forms, (ii) metacercariae in the fish, (iii) infected fish or (iv) intermediate host snails. There were significant differences between fish parasitized and not parasitized with M. appendiculatoides with respect to their DDT concentrations. There were also significant differences between the variances of the mean Clark's coefficient of condition values between catfish parasitized and not parasitized by M. appendiculatoides, with the variance of non-parasitized catfish being significantly larger. The results provided statistical evidence that DDT has a detrimental effect on M. appendiculatoides infection intensity. Furthermore, the significantly larger variance value of Clark's coefficient for non-parasitized fish suggested that DDT affects both the parasite and general host condition.

Despite the commercial and zoonotic importance of larval anisakid infestations of teleosts, their distribution among Australia's diverse marine fish fauna is poorly understood. A preliminary survey of Australia's tropical north-west revealed a generally high prevalence of larval anisakids representing four genera (Anisakis, Terranova, Thynnascaris and Raphidascaris) among only seven fish species. The potential impact of high larval anisakid infections on both the health of recreational fishermen and aquaculture environments is discussed.

During the summer and autumn of 1994, 1995 and 1996, 406 juvenile herring caught off the Swedish west coast were examined for parasites. Amongst those found was the digenean Pseudobacciger harengulae, which represents new host and locality records for this parasite. Pseudobacciger harengulae has been reported from several species of clupeiformes, mostly from tropical and temperate regions of the Atlantic, Pacific and Indian Oceans. The morphology of P. harengulae is described and compared with earlier descriptions of P. harengulae and P. manteri. The possible relationships between P. harengulae and P. manteri are discussed and the validity of the P. manteri is questioned. Most of the specimens (75%) of P. harengulae were found in the pyloric caeca and the remainder (25%) in the intestine.

In vitro released products of adult Setaria cervi females, microfilariae and extracts showed considerable amounts of collagenase activity. On the basis of per mg protein released in vitro, the products of both microfilariae and adult females exhibited comparable activity but this was much higher than that of extract of microfilariae and adult females. Two collagenase enzymes with molecular masses of 50 kDa and 70 kDa were separated using DEAE-sepharose CL6B and Sephadex G-100 column chromatography. The 50 kDa and 70 kDa collagenase exhibited pH optima of 5.2 and 7.0, respectivly. Considering specific activity, the 50 kDa enzyme was found to contribute about ten times more collagenase activity as compared to the 70 kDa enzyme. An inhibition study revealed obvious differences between them. Thiol group inhibitors such as N-ethylmaleimide and leupeptin inhibited the 50 kDa enzyme but this was strongly activated by dithiothreitol, a thiol group stabilizer. Alternatively, the 70 kDa enzyme showed a sensitivity to a metal chelator and a serine group inhibitor indicating its metalloserine protease nature. The antifilarial drug diethylcarbamazine did not demonstrate any inhibition under in vitro conditions. Both enzymes were significantly inhibited by antibody IgG separated from Wuchereria bancrofti infected human sera, showing a possible immunoprotective role.

Twenty European domestic cats were each infected with 15,000 protoscoleces of Echinococcus multilocularis extracted from metacestodes grown in experimentally infected common voles (Microtus arvalis). Sixteen days after infection, ten cats were treated with a broad-spectrum anthelmintic and acaricide comprising praziquantel and milbemycin oxime. Five days later treated and untreated cats were euthanized and the intestine examined for E. multilocularis. Five of ten untreated cats were infected with E. multilocularis with worm burdens ranging from 235 to 1920 worms per cat. No E. multilocularis were recovered from any of the treated cats. This study has demonstrated that this new combination broad spectrum anthelmintic and acaricide for cats is highly efficacious against E. multilocularis and the relevance of this is discussed.

Between 1993 and 1997, 837 stray dogs from North Taiwan were necropsied and examined for dog heartworm infections. A thick smear from 20 ml of peripheral blood from each dog was also prepared and examined for microfilariae (mf). The overall prevalence of adult worms in the dog population was 57%. The prevalence of mf in 1228 house dogs from different parts of Taiwan was also determined from 20 ml of peripheral blood in the same way. The overall prevalence of mf was 25%, with a value of 30% in the main island of Taiwan, this being 15 times higher than that in the offshore islands (2%). In Taiwan, the prevalence ranged from 4% in Hualien County, East Taiwan, to 41% in Nantou County, Central Taiwan. The mf prevalences on offshore islands were 1% on Liuchiu and 2% on Lanyu. The mf density per 20 ml blood in 82 house dogs was found to be 23 mf per dog, with a range of 3–97 mf per dog. A total of 477 stray dogs were found to be infected with adult worms of Dirofilaria immitis. The mean number of 7 worms per dog was obtained, with a range of 1–55 worms per dog. These results indicate that the prevalence of canine dirofilariasis has increased in Taiwan over the past 10 years. Moreover, the prevalence may be related to the wind speed, temperature, relative humidity, and altitude in the different areas surveyed.

Two male worms of Camelostrongylus mentulatus were found in one of 84 murciano-granadina goats in Murcia, south-eastern Spain. This is the first report of C. mentulatus in goats in the Iberian peninsula and the possible origin and transmission of this nematode in Spain are discussed.

It is demonstrated that Schistosoma curassoni, a parasite of sheep, cattle and goats in parts of West Africa, will live for at least 8 years 5 months in a sheep. The sheep was exposed to 500 cercariae of S. curassoni liberated from infected Bulinus wrighti. The sheep died of natural causes, and at post-mortem 28 pairs of adult S. curassoni were removed from the mesenteric and rectal veins. All female worms were gravid, and eggs were hatched from faeces to produce miracidia. The development of immune responses of the host had apparently little or no effect on the viability of the eggs. Histological studies of the liver, small and large intestines revealed mild pathological symptoms. The longevity of S. curassoni is the first record of longevity of schistosomes to be based on worm counts.