The epidemiology of Ancylostoma spp. was studied in the endangered Iberian lynx (Lynx pardinus) in the Doñana National Park, south-west Spain. Faecal samples were collected throughout a complete annual cycle (August 1997 to September 1998). The overall egg prevalence of Ancylostoma spp. was 57.8%. The pattern of abundance of Ancylostoma spp. eggs in faeces was overdispersed. Juvenile lynx demonstrated a statistically higher prevalence and abundance of Ancylostoma spp. than in adults. These levels of egg output (maximum 21195 epg), as previously reported in free ranging large felid cubs, could be close to disease involvement. The potential pathogenicity of hookworms and the influence of individual and ecological factors on hookworm transmission in the Iberian lynx from the Doñana National Park population are discussed.

1. B. glabrata exposed to the sublethal concentrations (0·04 ppm, 0·03 ppm and 0·02 ppm) of TPLA died from the effects of the molluscicide over a period of time. Most of the deaths occurred between the 10th and 35th days after treatment, showing that although TPLA is slow acting, its effect is not continuous but restricted to a limited period of time.

2. 0·01 ppm of TPLA was chosen as a sublethal concentration to study the effect of the molluscicide on the development of S. mansoni in B. glabrata.

3. 240 B. glabrata were infected with S. mansoni, and 40 of these were treated with 0·01 ppm of TPLA 4 days before infection. The remaining 200 were divided into 5 groups of 40 each, one group being maintained as a control and the other four treated 2, 10,15 and 20 days postinfection respectively. Other suitable controls for comparison were also maintained.

4. The infection rate in all groups was high, and comparison with the control showed that there was no significant difference in infection rate between the treated groups and the control group at the 5% level. However, the infection rate in the preinfection treated group was lower than in the control group and significant at the 10% level.

Haemonchus contortus and Haemonchus placei are among the major parasites of small ruminants and cattle. Although infection with these nematodes is host-specific, with H. placei predominating in cattle and H. contortus in sheep, cross-infections are observed in areas where both parasites are sympatric, and hybrid offspring can occur. Therefore, a fast and precise method is required for differentiating the parasites. Identification based on spicule morphometry is the most common technique for differentiating Haemonchus species. However, because these measurements overlap between species, morphological analysis is insufficient for differentiating between helminth species. In this work, we present a reliable, conventional polymerase chain reaction (PCR)-based method that uses two species-specific primer pairs to differentiate between H. contortus and H. placei specimens and their hybrids. Each primer pair produces one single and distinct amplification band for each species, which enables the detection of hybrid specimens. These primer pairs were validated by testing eight different populations of H. contortus, H. placei and hybrids.

Sixty-one serum samples selected on the basis of reactivity in the complement fixation (CF) and latex agglutination (LA) test, were further examined for sensitivity and specificity by indirect haemagglutination (IHA), enzyme linked immunosorbent assay (ELISA) and defined antigen substrate spheres (DASS). Twenty sera from healthy Europeans and 48 samples from patients with either schistosomiasis or trichinosis were also tested. Comparable levels of sensitivity were found between the CF and LA positive sera and IHA, ELISA and DASS. Of the CF positive LA negative group of sera, many were positive by DASS but only a few reacted in IHA and ELISA. Some cross reactivity was also observed in the schistosomiasis sera tested by IHA and ELISA.

A cross-sectional survey was conducted to determine the prevalence and intensity of soil-transmitted helminths among pre-school and school-aged children attending nursery and primary schools in Ile-Ife. Single stool samples were collected between January and March, 2009 from 352 children randomly selected from a total of 456 children attending both private and government schools. The stool samples were processed using the modified Kato–Katz technique, and then examined for the eggs of soil-transmitted helminths (STHs). One hundred and twenty-one (34.4%) samples were positive for STH eggs. The overall prevalences of Ascaris lumbricoides, Trichuris trichiura and hookworm were 33.2%, 3.7% and 0.9%, respectively. The prevalence of STH infection in government schools (47.8%) was significantly higher than in private schools (16.1%) (P < 0.001). The most common type of mixed infection was the combination of A. lumbricoides and T. trichiura (6.8%). The prevalence and intensity of A. lumbricoides rose with age. The lowest prevalence and intensity (7.7%; 0.240 ± 0.136 eggs per gram (epg)) were recorded in the 2- to 3-year-old age group, while the highest prevalence and intensity (58.7%; 1.820 ± 0.237 epg) were recorded in children aged 10 years and above. A questionnaire survey indicated that 73% of the children attending private school had been treated with anthelminthics less than 2 months prior to the collection of stool specimens, while 43% of the children attending government school received anthelminthic treatment during the same period. The findings indicate that STH infections are endemic among schoolchildren in Ile-Ife and that the burden of parasitic infections is greater in government schools than in private schools.

The life cycle of Genarchopsis goppo a hemiurid trematode found in the stomach of Channa punctata has been worked out in detail. The egg contains a fully developed miracidium at the time of liberation. The miracidium contains a ciliated covering, a long apical gland and a crown of spines at the anterior end. The snail Amnicola travancorica acts as the first intermediate host inside which the miracidium passes through sporocyst and redial generations. The cercaria is of cystophorous type and is identical to Cercariae Indicae Sewell XXXV. Metacercaria occurs in the ostracods Stenocypris malcolmsoni and Eucyoris capensis. The fish Aplocheilus panchax serves as the paratenic host. The entire developmental cycle from egg to egg producing adult takes 3 months.

We estimated the prevalence of Dirofilaria immitis infection in domestic dogs in five Turkish provinces – Sakarya, Kocaeli, Ankara, Elazig and Mersin – using a commercial ELISA kit for detecting circulating antigen and a PCR test for detecting circulating microfilarial DNA. A total of 211 whole-blood and serum samples were collected from dogs of various breeds, ages and life status (owned or stray). Sample population characteristics were recorded and examined for differences in prevalence. Additionally, we collected 15 blood samples from cats (14 owned and 1 stray) from Ankara province and used PCR to detect D. immitis infection. Twenty-seven (12.8%) of 211 dog samples were positive for D. immitis antigen by ELISA. No differences in prevalence were observed by sex (female: 14.4%; male: 10.7%; P>0.05). The prevalence of D. immitis infection varied with age: 11.8% in younger dogs (0.5–2 years) and 17.5% in older dogs (3–5 years). Prevalence between stray dogs (15.2%) and owned dogs (9.3%) did not differ (P>0.05). Prevalence rates were highest in Kocaeli province (18.3%), followed by Ankara (14.8%), Sakarya (12.3%) and Mersin (10.5%) provinces. Prevalence in Elazig province was 0%. No dogs or cats had microfilarial DNA detectable by PCR.

Soil contaminated with helminth eggs and protozoan cysts is a potential source of infection and poses a threat to the public, especially to young children frequenting playgrounds. The present study determines the levels of infection of helminth eggs in soil samples from urban and suburban playgrounds in five states in Peninsular Malaysia and identifies one source of contamination via faecal screening from stray animals. Three hundred soil samples from 60 playgrounds in five states in Peninsular Malaysia were screened using the centrifugal flotation technique to identify and determine egg/cyst counts per gram (EPG) for each parasite. All playgrounds, especially those in Penang, were found to be contaminated with eggs from four nematode genera, with Toxocara eggs (95.7%) the highest, followed by Ascaris (93.3%), Ancylostoma (88.3%) and Trichuris (77.0%). In addition, faeces from animal shelters were found to contain both helminth eggs and protozoan cysts, with overall infection rates being 54% and 57% for feline and canine samples, respectively. The most frequently occurring parasite in feline samples was Toxocara cati (37%; EPG, 42.47 ± 156.08), while in dog faeces it was Ancylostoma sp. (54%; EPG, 197.16 ± 383.28). Infection levels also tended to be influenced by season, type of park/playground and the texture of soil/faeces. The occurrence of Toxocara, Ancylostoma and Trichuris eggs in soil samples highlights the risk of transmission to the human population, especially children, while the presence of Ascaris eggs suggests a human source of contamination and raises the issue of hygiene standards and public health risks at sites under investigation.

This study aimed to evaluate the helminth parasites of Geophagus proximus from the São José dos Dourados River, a tributary of Paraná River, Ilha Solteira Reservoir, São Paulo State, Brazil. From May 2006 to May 2007, 116 G. proximus specimens were examined and seven different taxa of helminth were collected and identified: proteocephalidean plerocercoids (Cestoda); metacercariae of Austrodiplostomumcompactum, Clinostomum heluans and Clinostomum sp. (Trematoda); and Raphidascaris (Sprentascaris) hypostomi, and larvae of Raphidascaris sp. and Contracaecum sp. (Nematoda). All parasites presented the typical aggregated pattern of distribution, as well as the presence of a high number of larval stages, an absence of influence of the host sex and seasonality upon community parameters, as well as a correlation between species richness and host body weight. Moreover, with the exception of A. compactum metacercariae, all helminths found in this study are reported for the first time in G. proximus.

Hormones play a significant role in murine cysticercosis (Taenia crassiceps), and increase the frequency of porcine cysticercosis caused by Taenia solium. In the present study, we report the in vitro effect of insulin on the larval stages of T. crassiceps (ORF strain) and T. solium. In vitro exposure of T. crassiceps cysticerci to insulin was found to stimulate this parasite's reproduction twofold with respect to control values, while the same treatment had no effect on T. solium cysticerci. Moreover, normal female mice (BALB/cAnN) infected with T. crassiceps cysticerci previously exposed to insulin presented larger parasite loads than mice inoculated with vehicle-treated cysticerci. To determine the possible molecular mechanisms by which insulin affects T. crassiceps, the insulin receptor was amplified by means of reverse transcriptase-polymerase chain reaction (RT-PCR). Interestingly, both T. crassiceps and T. solium expressed the insulin receptor, although insulin had effects only on T. crassiceps. These results demonstrate that insulin has a dichotomistic effect; it acts directly only on T. crassiceps cysticerci reproduction, possibly through its binding to a specific insulin receptor synthesized by the parasite. Thus, insulin may be recognized by T. crassiceps cysticercus cells as a mitogenic factor, and contribute to parasite proliferation inside the host, as well as to the female mouse susceptibility to T.crassiceps. This phenomenon has not been reported for cysticercosis caused by T. solium, which could, in part, be related to the poor effect of insulin upon the human parasite.

The genetic mechanisms of how free-living nematodes evolved into parasites are unknown. Current genetic model nematodes (e.g. Caenorhabditis elegans) are not well suited to provide the answer, and mammalian parasites are expensive and logistically difficult to maintain. Here we propose the terrestrial gastropod parasite Phasmarhabditis hermaphrodita as a new alternative to study the evolution of parasitism, and outline the methodology of how to keep P. hermaphrodita in the lab for genetic experiments. We show that P. hermaphrodita (and several other Phasmarhabditis species) are easy to isolate and identify from slugs and snails from around the UK. We outline how to make isogenic lines using ‘semi-natural’ conditions to reduce in-lab evolution, and how to optimize growth using nematode growth media (NGM) agar and naturally isolated bacteria. We show that P. hermaphrodita is amenable to forward genetics and that unc and sma mutants can be generated using formaldehyde mutagenesis. We also detail the procedures needed to carry out genetic crosses. Furthermore, we show natural variation within our Phasmarhabditis collection, with isolates displaying differences in survival when exposed to high temperatures and pH, which facilitates micro and macro evolutionary studies. In summary, we believe that this genetically amenable parasite that shares many attributes with C. elegans as well as being in Clade 5, which contains many animal, plant and arthropod parasites, could be an excellent model to understand the genetic basis of parasitism in the Nematoda.

Experiments on the influence of egg density and varying quantities of chlamydospores on the nematode-trapping ability of Duddingtonia flagrans, influence of D. flagrans on the larval translation of gastrointestinal nematodes, doses of chlamydospores required for the effective control of gastrointestinal nematodosis and the epidemiology of nematode parasites were conducted in goats, which generated baseline data required for strategic application of the biocontrol agent. The nematode-trapping ability of D. flagrans, measured by numerical enumeration of infective third-stage larvae developed in the faecal culture, revealed that the efficacy is dependent on both nematode egg and chlamydospore density. Pasture plot studies revealed that D. flagrans, if deposited at the same time as nematode eggs, prevents translation of third-stage larvae of caprine nematodes from the faecal pats onto the grass blades. Feeding of 1 × 106 chlamydospores per kg body weight and above to goats virtually eliminated larvae from the pasture. Application of as few as 1 × 104 and 1 × 105 chlamydospores per kg body weight had a profound impact on larval recovery. The effect persisted as long as the chlamydospores were fed. Monthly faecal worm egg counts of adult goats maintained under a semi-intensive management system on the Chhattisgarh plain and pasture larval burden revealed that June to August were the months of high risk for nematodosis. Haemonchus was the dominant species recorded throughout the year. The present data can best be utilized by formulating a strategic control measure when the larval challenge to the animal is maximum (June to August), so as to prevent establishment of patent infection. The observations reinforced the strategy to be adopted for nematode parasite control in goats by applying the biocontrol option at the onset of the monsoon.

Onchocerca tarsicola is a subcutaneous filariid of the red deer living on the tendons of the radial-carpal and tibial-tarsal joints. Its micronfilariae concentrate mainly in the external ears. The extrinsic cycle of this parasite was investigated. In a hyperendemic area wild-caught Odagmia ornata were dissected, as well as samples which had fed on a tame deer and been kept in the laboratory after the infecting blood-meal. About 3 to 5% of the flies were infected with larval stages of Onchocerca tarsicola. More than 40% of Odagmia ornata induced to feed on the ears of the deer became infected and about 30% of the flies which survived more than 18 days contained third-stage larvae. The larval development in the vector is described. Prosimulium nigripes appears to be a second natural vector of Onchocerca tarsicola. Observations concerning the potential vectors of O. Jlexuosa, O. tubingensis and Cutifilaria wenki living in red deer are discussed.

Tryptic-like enzymes have been extracted from the intestines and intestinal contents of Ascarts lumbricoides and Strongylus edentatus. The action of these enzymes, in relation to hydrogen ion concentration, has been examined.

The parasite trypsins resembled pancreatic trypsin in that the relative production of “free acid” and “formaldehyde acid” was similar, but though acting only on the alkaline side of the isoelectric points of the substrate proteins, optimum action on gelatin, blood-albumin and casein was obtained at pH 6·2.

The amount of protease extracted from a given weight of S. edentatus tissue was far greater than that from a similar amount of A. lumbricoides. Thus Strongylus digested 4·9 to 8·3 times as much gelatin, 12·5 to 40·9 times as much casein and 2·5 to 5·2 times as much blood-albumin as Ascaris.

Spectroscopic examination of the process of digestion of oxyhaemoglobin showed, first, the formation of reduced haemoglobin and then the formation of haematin. These changes took place most rapidly at higher hydrogen ion concentrations.

Owing to the close association and proximity of man with his domestic animals (in particular the dog and cat), there exist the possibilities of human infection by certain helminth parasites of these animals which may, in some circumstances, lead to serious problems of public health, e.g. encephalitis and granulomatous lesions of the eye caused by toxocariasis (Ashton, 1960; Woodruff et al., 1964).

1. Eggs of Contracaecum osculatum, extracted from grey seal faeces, have been hatched in the laboratory at different temperatures and the morphology and behaviour of the larvae described.

2. Artificial exsheathment of the second stage larva from the first stage cuticle has been demonstrated.

3. In attempting to establish the first intermediate host of the life cycle, two species of harpacticoid copepods have been successfully infected but no significant developments reported.

The process of spermiogenesis in two paramphistomes, Sandonia sudanensis and Basidiodiscus ectorchis from the Nile fish Synodontis schall in Egypt was studied by transmission electron microscopy. Spermiogenesis is characterized by the outgrowth of the zone of differentiation, presenting two basal bodies separated by a microtubule organizing centre, each basal body developing into a flagellum. Proximodistal fusion of these flagella with a median cytoplasmic extension gives rise to the spermatozoon. The mature spermatozoon possesses two axonemes of the 9+‘1’ pattern typical of parasitic helminths. There are few ultrastructural studies on spermiogenesis in paramphistomes, which are considered the most primitive digenetic trematodes. The present study provides new and more detailed information on this process, including the presence of a lateral flange and external ornamentation of the cell membrane. The value of sperm ultrastructure as a taxonomic tool in phylogeny is also discussed.

Diphtherostomum brusinae was first recorded by the present study in the north of Portugal. Sporocysts, containing cercariae and encysted metacercariae, were observed in the gonads and digestive gland of the gastropod Nassarius reticulatus. Metacercariae were also found infecting the foot, mantle border and gills of the cockle Cerastoderma edule. The adult form was lodged in the rectum of the definitive host Diplodus sargus. The morphology of the three parasitic stages was studied by light (LM) and scanning electron microscopy (SEM). Despite the close similarity between cercaria and metacercaria, SEM data provided information that allowed their differentiation, namely the presence of a dense crown of microvilli around the oral cavity of the cercariae, which was absent in the metacercariae. In addition, the metacercariae presented a specific pre-acetabular rectangular band with conspicuous triangular spines. The adult showed characteristics of D. brusinae species, in particular the presence of acetabular lips, compact vitellaria and large elliptical eggs. Sequenced ITS1 data clearly demonstrated that the cercariae and metacercarial cysts from N. reticulatus, the cysts from C. edule and the adult isolated from D. sargus were life cycle stages that belonged to the same species, i.e. D. brusinae. Two transmission strategies in the life cycle of this species were observed: (1) cercariae encyst within the sporocysts of N. reticulatus and await ingestion by the definitive host; and (2) N. reticulatus naturally emits cercariae; they encyst in C. edule or the environment and are ingested by the definitive host.

Different species of Cassia plant are widely available in India and are commonly used either for their laxative, antimicrobial or antibacterial activity. In the present study the effectiveness in vitro of the crude alcoholic extracts of three species, namely Cassia alata, C. occidentalis and C. angustifolia, in the early paralysis and mortality of the fowl tapeworm Raillietina tetragona at concentrations ranging from 5 to 80 mg/ml was investigated. Time of paralysis and death were monitored frequently. Immediately after paralysis the tapeworms were processed for electron microscopic studies. While the untreated or control parasites survived for 81.93 ± 5.85 h, the parasites treated with C. alata took less time (1.68 ± 0.27 h) to be paralysed, followed by those treated with C. angustifolia (2.95 ± 0.29 h). Although C. occidentalis took more time (4.13 ± 0.31 h) to paralyse, in combination with either C. alata or C. angustifolia the time taken to paralyse became shorter. All the plant-treated parasites showed irrevocable changes in the scolex and proglottids as compared with the control, and these observations are comparable with those obtained with praziquantel. These results indicate that the three plants tested can be claimed to have anthelmintic activity in addition to their known properties, both when used individually and in combination. Further investigations will be required to evaluate their mechanism of action.