Fifteen lots of fifty individual cysts from three separate batches were exposed to the action of potato root diffusate, the hatching allowed to continue to completion and the total larvae liberated from each cyst were counted. The data obtained were analysed both as they stood and after three different transformations for variability, as well as for independence between mean and variance. Very high variability corresponding to a coefficient of variation of 1.00 was detected, and there was, moreover, a very significant correlation between mean and variance. The most suitable transformations appeared to be either logarithmic or angular—the latter being probably the more useful. The latter, moreover, tended to increase the sensitivity of analysis. The importance of residual counts when hatching is complete is emphasized even in the absence of an angular transformation of the data.

Laboratory experiments were performed to investigate the ability of isolates from the predacious fungus species Arthrobotrys conoides (one isolate) and A. robusta (four isolates) to trap and kill infective Haemonchus placei larvae. Three groups for each isolate were formed: group 1, fungi and infective larvae; group 2, fungi; group 3, infective larvae. There were statistically significant differences (P<0.05) between the antagonistic effects of three isolates of A. robusta, but there was no such difference between the isolate of A. conoides and isolates of A. robusta. This result indicates that there can be a variation in the predatory capacity of different isolates within a single fungus species. After 20 days of assay, there was greater recuperation of infective H. placei larvae in group 3 compared to group 1.

Adults of Clinostomum spp. are digenetic trematodes found in fish-eating birds, reptiles and occasionally mammals, including humans. Freshwater snails serve as first intermediate hosts and many fish species and amphibians as second intermediate hosts. To date, amphibian hosts of Clinostomum metacercariae include members of urodele and anuran families in North America, but no data are available on infections of European amphibians, including newts. In this study, we characterize infections of Clinostomum complanatum metacercariae in four smooth (Lissotriton vulgaris) and 18 Italian crested newts (Triturus carnifex) from an artificial pond located in a protected area in Tuscany, Italy. Parasites were surgically removed from the infected newts and identified both morphologically and using sequences of a mitochondrial gene, cytochrome c oxidase I, and the ribosomal markers, internal transcribed spacers. This is the first record of C. complanatum in European newts and, more generally, in amphibians in Europe.

A monitoring programme aimed at the diagnosis of subcutaneous dirofilariasis and heartworm disease in working (police and military) dogs in Slovakia has been performed during the period of September 2007 to February 2008. In co-operation with the Ministry of the Interior and the Ministry of Defence, in total, 710 dogs (591 police dogs and 119 military dogs) were investigated for the presence of microfilariae in blood. All police and military dogs in active service held on the territory of Slovakia were included. Microfilariae were detected in 118 (20.0%) police dogs and 10 (8.4%) military dogs. The most infected individuals originated from southern parts of Slovakia (Trnava region 53.6% and Nitra region 39.6%); the prevalence was low in northern regions (Žilina 3.1% and Prešov 6.6%). In several districts of southern Slovakia, the prevalence of subcutaneous dirofilariasis in working dogs exceeded 40%. In all infected animals, the autochthonous origin of the disease was confirmed; however, due to the frequent movement of working dogs, it was not possible to identify the exact locality of infection. At present, a dog living in Nemšová village in Trenčín district (north-western part of the country) is regarded as the northernmost localized autochthonous case of subcutaneous dirofilariasis in Slovakia. In three dogs, co-infection of Dirofilaria repens and Dirofilaria immitis was detected. High prevalence rates in working dogs and the zoonotic characteristic of the disease represent an undoubtedly important veterinary and medical problem that requires the urgent introduction of prophylactic and control measures.

A new species, Paradiplozoon ichthyoxanthon (Monogenea: Diplozoidae) is described from the gills of Labeobarbus aeneus (Cyprinidae) in the Vaal Dam and Vaal River Barrage in the Vaal River, South Africa. This species is the first in this genus from southern Africa; Paradiplozoon aegtyptense and Paradiplozoon ghanense have been described from northern Africa. The new species differs from all other Paradiplozoon species on the basis of the combination of morphological characteristics, namely disk-like haptor, eggs without filaments, clamp and central hook morphology and size of specimens. The species description is accompanied by a description of the reproductive organs. The internal transcribed spacer of the ribosomal gene (ITS2) was studied and the South African individuals formed a distinct clade, supporting the establishment of a new species.

The presence of helminths and protozoans in public squares and lawns of the city of Maringá, southern Brazil, during winter and summer was assessed in order to evaluate their seasonal fluctuations in relation to edaphic and climatic factors. Samples were collected from January 2003 through June 2004 in 90% (13) of all public squares covered by sand, and in 30% (4) of all lawns used as leisure areas. The samples were analysed quantitatively by modified centrifugal-flotation and sedimentation in water techniques, and qualitatively by a method based on positive larval thermo-hydrotropism. Meteorological data were recorded, and physical, chemical and structural characteristics of the soil were analysed. One hundred and thirty samples of sand from squares, 65 in summer and 65 in winter, and 40 samples of grass from lawns, 20 in each season, were collected. All samples from lawns, 62 (95.38%) from squares in winter and 45 (69.23%) in summer, contained protozoans and/or helminths. Eggs of Toxocara spp. were the most frequently observed parasites in both winter and summer in squares (P < 0.0001) and in lawns (P = 0.6142), being equally distributed among the different locations (P = 0.2038). Species diversity was lower in winter; fewer parasites were found in summer. This region, with a tropical climate and a mild winter dry season, has favourable edaphic and climatic conditions for soil contaminants to persist year-round. In addition, the high frequency of animals such as dogs and cats and the poor sanitary measures in force made it possible for zoonoses to be transmitted in the public spaces.

This review analyses the scarce available data on biodiversity and transmission of helminths in Arctic coastal ecosystems and the potential impact of climate changes on them. The focus is on the helminths of seabirds, dominant parasites in coastal ecosystems. Their fauna in the Arctic is depauperate because of the lack of suitable intermediate hosts and unfavourable conditions for species with free-living larvae. An increasing proportion of crustaceans in the diet of Arctic seabirds would result in a higher infection intensity of cestodes and acanthocephalans, and may also promote the infection of seabirds with non-specific helminths. In this way, the latter may find favourable conditions for colonization of new hosts. Climate changes may alter the composition of the helminth fauna, their infection levels in hosts and ways of transmission in coastal communities. Immigration of boreal invertebrates and fish into Arctic seas may allow the circulation of helminths using them as intermediate hosts. Changing migratory routes of animals would alter the distribution of their parasites, facilitating, in particular, their trans-Arctic transfer. Prolongation of the seasonal ‘transmission window’ may increase the parasitic load on host populations. Changes in Arctic marine food webs would have an overriding influence on the helminths’ circulation. This process may be influenced by the predicted decreased of salinity in Arctic seas, increased storm activity, coastal erosion, ocean acidification, decline of Arctic ice, etc. Greater parasitological research efforts are needed to assess the influence of factors related to Arctic climate change on the transmission of helminths.

A total of nine playground sites in Dublin city and county were surveyed from which 228 samples were removed. Of these samples, 15% were positive for Toxocara canis ova and a mean egg density of 1.4 ova/100 g was calculated. Two types of playground sites were identified and that the level of toxocaral contamination was found to be greater in ‘neighbourhood playgrounds’ than in ‘adventure playgrounds’. There was no significant difference in the number of positive samples taken from sites both inside and outside these playgrounds. No evidence of Toxocara cati ova was found. Only one sample was positive for Toxascaris leonina ova. Over 50% of the T. canis ova identified were infective. The findings in this study suggest that the sample's moisture content contributes to the long term survival of infective Toxocara ova in the environment. A recovery rate of 69.8% was achieved with a modified version of the flotation method.

The community structure exhibited by strongyloid nematodes from the large intestines of horses was examined using data from autopsies of 150 horses. Thirty-one species of nematodes were encountered, but they were not clearly divisible into core and satellite species. Multiple congeneric, consubfamilial and confamilial species were a prominent feature of the community and were more common than singleton infections. Multivariate analyses provided evidence of a stable community of helminths dominated by positive interactions but with few negative interactions, suggesting the absence of competition within the community.

Computer-assisted microscopy and multivariate statistics were used to establish and evaluate a procedure for identification of bovine strongylid eggs. Ostertagia ostertagi, Cooperia oncophora, Haemonchus placei, Trichostrongylus axei, and Oesophagostomum radiatum eggs were obtained from faeces voided by monospecifically infected calves. Images of single eggs (400× magnification) were recorded by a CCD camera fitted onto a microscope and digitizied on a PC. After separation of eggs from the image background, the pixel (picture element) positions of the egg outline were analysed by algorithms to describe size and shape. A stepwise discriminant analysis was subsequently used to select and rank descriptive features of 4207 eggs according to discriminatory power. Classification criteria were developed by linear discrimination analysis on the basis of selected features, and the criteria evaluated by cross-validation. A maximum average percentage of correct classification of 85.8% resulted when nineteen features were employed in a linear classification criterion. The percentages correct classification for each species were: O. ostertagi 76.3%, C. oncophora 90.8%, O. radiatum 87.8%, H. placei 90.1%, and T. axei 83.8%. Classification based on the five most important features gave an overall correct classification of 81.5%. Images of ‘unknown’ eggs could be identified automatically by the classification criteria after procedural steps performed by PC were linked in a batch program.

Fasciola hepatica has recently emerged as a major pathogen of humans from reports on areas of endemicity and hyper-endemicity for fascioliasis. This situation is aggravated by the lack of standard assays for the screen diagnosis of F. hepatica infection in humans living in endemic areas. Our laboratory has developed an enzyme-linked immunosorbent assay (Fas2-ELISA) based on the capture of IgG antibody by a purified protein Fas2, which is an adult fluke cysteine proteinase. Fas2-ELISA exhibited 95% sensitivity and 100% specificity in 38 individuals infected with F. hepatica diagnosed by finding eggs in stools and 46 serum samples from healthy volunteers. No cross-reaction was observed with 54 serum samples from patients with ten different parasitic infections including the trematodes Paragonimus westermani and Schistosoma mansoni. The high antigenicity of Fas2 is suggested by the fact that antibodies to Fas2 rise rapidly by 1–2 weeks of infection and rise until patency at 8 weeks of infection in experimentally infected alpacas. Field screening for human fascioliasis using Fas2-ELISA and coprology in three endemic locations of the Peruvian Andes resulted in 95.5% sensitivity, 86.6% specificity in a population of 664 children in an age range of 1 to 16 years old. These results provide evidence of the clinical potential of Fas2-ELISA to diagnose fascioliasis in humans exposed to liver fluke infection in endemic areas for this parasite. Fas2-ELISA is currently developed as a standard assay for both field screening for fascioliasis in people living in endemic areas and detecting occasionally F. hepatica infected patients in clinical laboratories.

Metacercariae of the trematode Curtuteria australis (Echinostomatidae) accumulate in the foot of the New Zealand cockle Austrovenus stutchburyi, severely impairing the cockle's ability to burrow under the sediments. This results in increased predation by birds on cockles, and thus enhanced transmission rates of the parasite to its bird definitive hosts. This host manipulation by the trematode is costly: fish regularly crop the tip of the foot of cockles stranded on the sediment surface, killing any metacercariae they ingest. A second, previously undetected trematode species (characterized by 23 collar spines) co-existing with C. australis, has been found in the foot of cockles in the Otago Harbour, South Island, New Zealand. The relative abundance of the two species varies among localities, with the identity of the numerically dominant species also changing from one locality to the next. Both C. australis and the new species have a strong preference for encysting in the tip of the cockle's foot, where their impact on the burrowing ability of the host is greatest, and where they both face the risk of cropping by fish. Results indicate that these two species are ecological equivalents, and their combined numbers determine how the cockle population is affected.

Abstract A total of 475 soil samples were collected from five public park playgrounds, 17 kindergarten sandpits and 124 housing estates in Resistencia, a medium-sized subtropical-region city in Argentina, and processed by the centrifugal flotation method. Eggs of Toxocara spp. were present in five (3.4%) of the 146 habitats surveyed and in six (1.3%) of the 475 samples examined. Twenty per cent of public parks, 5.9% of kindergarten sandpits and 2.4% of housing estates were contaminated with Toxocara eggs. Depending on the number of samples examined from the three types of habitat, contamination by Toxocara was 0.7% in public park playgrounds, 1.2% in kindergarten sandpits and 1.6% in the housing estates. High prevalences of Ancylostomidae eggs were also found especially in public park playgrounds with a value of 100%, compared with 19.4% found in housing estates and 11.8% in kindergarten samples. These results suggest that in Resistencia, human infections with Toxocara are likely to occur within the limits of housing estates more so than in public parks or open spaces.

Samples of wild rabbits were collected each month over the six-year period 1977–82 and their intestines and abdomens examined for helminths. Three species of nematode, four of cestode and one trematode were found, Graphidium strigosum was recorded in 33% of the 786 rabbits, Trichostrongylus retortaeformis in 79%, Passalurus ambiguus in 14·2% Cittotaenia Pectinata in 37%, C. denticulata in 11%, Coenurus pisiformis in 3%, Cysticercus serialis in 0·1% and Fasciola hepatica in 0·1%. G. strigosum and P. ambiguus infections were greatest in the older heavier animals whereas juvenile rabbits had the heaviest T. retortaeformis nematode burdens. Seasonal fluctuations were observed in G. strigosum, T. retortaeformis, P. ambiguus and Cittotaenia denticulata. The sex and reproductive state of the female rabbit did not have any significant effect on either the nematode or cestode worm burdens. Myxomatosis was shown to significantly increase the worm burden of T. retortaeformis and raise that of P. ambiguus. No nematode or cestode from domesticated animals were recovered and it was concluded that since F. hepatica was found only once rabbits did not constitute a serious hazard to the farm animals grazing the area.

The present study aimed to investigate the seasonal epidemiological prevalence of gastro-intestinal nematodes (GINs) of goats with respect to sex and age of the host in the Kashmir valley from 1 February 2005 to 31 January 2007. A total of 1267 goats were examined [faecal examination: 938 (year 1: 470; year 2: 468); gastro-intestinal (GIT) examination: 329 (year 1: 175; year 2: 154)]. The overall prevalence of GIN infection in these animals was 54.3% (year 1: 54.8%; year 2: 53.8%; P = 0.842). The different parasites reported with their respective prevalences (%) were: Haemonchus contortus (48.3); Bunostomum trigonocephalum (30.1); Chabertia. ovina (29.8); Ostertagia circumcincta (29.8); Nematodirus spathiger (25.2); Trichostrongylus spp. (25.1); Oesophagostomum columbianum (23.5); Trichuris ovis (19.0); and Marshallagia marshalli (16.6). The mean maximum prevalence of GIN infection (faecal examination: 75.6 ± 0.20; GIT examination: 85.3 ± 0.95), faecal egg counts (2552 ± 85.7) and average worm burden (333.25 ± 2.25) were found in the summer and they were lowest in winter (prevalence: faecal examination, 23.2 ± 0.95; GIT examination, 12.7 ± 0.20; faecal egg counts: 134.15 ± 9.15; and average worm burden: 79.8 ± 52.2), with significant differences between the seasons (P < 0.05). The sex of the hosts was not an important factor influencing the prevalence of GIN infection. With the increase in host age, prevalence of infection decreased significantly (P ≥ 0.05). Thus seasonal dynamics and age of the host animals significantly influenced the prevalence of GIN infection. The above findings will be helpful in devising the appropriate control strategies for GINs of goats reared under the traditional husbandry system in temperate agro-climatic conditions in the Kashmir valley as well as in similar climatic zones of other parts of the world.

During a non-targeted survey for entomopathogenic nematodes in South Africa, a new species of Steinernema was isolated from a soil sample collected from underneath a guava tree, close to the shore at Jeffrey's Bay. The nematode was isolated by means of the insect-baiting technique using last-instar larvae of Galleria mellonella. It is described herein as Steinernema jeffreyense n. sp. The nematode can be separated from other described, closely related species in terms of the morphological and morphometric characteristics of the different life stages, and in terms of the characterization and phylogeny of DNA sequences of the internal transcribed spacer (ITS) rDNA of the 18S gene, and of the D2D3 region of the 28S rDNA gene. The new species is placed molecularly in the arenarium–glaseri–karii–longicaudatum group characterized by the following morphological characters: infective third-stage juvenile with a body length of 926 (784–1043) μm, distance from head to excretory pore of 87 (78–107) μm, tail length of 81 (50–96) μm, with an E% of 109 (86–169), and eight evenly spaced ridges (i.e. nine lines) in the middle of the body. First-generation males have a spicule length of 88 (79–95) μm and gubernaculum length of 57 (51–61) μm. Male mucron is absent in both generations. First-generation females have an asymmetrical protuberance and a short, double-flapped epiptygmata, with both flaps directed to the front. The tail of the first-generation female is shorter than the anal body width, with a mucron on the dorsal tail tip, with D% = 78 (59–99). Cross-hybridization with S. khoisanae, S. tophus and S. innovationi showed the new species to isolate reproductively from the others. The analyses of ITS rDNA and D2D3 sequence of the 18S and 28S rDNA genes support the studied nematode isolate to be a valid new species belonging to the ‘glaseri’ group (Clade V).

The lack of a rapid and reliable method of making permanent preparations of nematodes is most strongly felt in the case of the plant-parasitic forms. The method in general use for most plant- and soil-inhabiting nematodes consists in relaxing the worms by gentle heat, fixing in 5% formalin or Ditlivsen's fixative (Thorne, 1925) and then impregnating slowly with glycerine. This procedure is quite reliable for the non-plant-parasitic forms but its results with plant-parasitic worms are very uncertain. With the latter it is necessary to transfer from fixative to very dilute glycerine, usually 1.5% glycerine in 7.5% alcohol with a trace of thymol or copper sulphate to discourage moulds. The glycerine is allowed to concentrate very slowly during at least four weeks until the worms can be transferred to pure glycerine in which they are mounted. Even then the results are often disappointing for the worms very frequently collapse.

In considering this general problem recently the authors called to mind the appearance of worms stained within plant tissues by the acid fuchsin-lactophenol method (Goodey, 1987: Franklin, 1949). This method causes no collapse or distortion of the worms and takes but a few minutes from living material to permanently stained preparations. The process has now been developed for use with free specimens of Anguillulina dipsaci and plant-parasitic species of Aphelenchoides, and is as follows:

The helminth fauna of two sympatric congeneric skinks (Mabuya agilis and M. macrorhyncha) from two distinct ‘restinga’ habitats (Praia das Neves and Grussaí) in southeastern Brazil were studied, totalling four data sets (sample sizes ranging from 11 to 28). A total of ten helminth species were associated with the skinks: Raillietiella sp., Paradistomum parvissimum, Pulchrosomoides elegans, Oochoristica ameivae, Hexametra boddaertii, parapharyngodon sceleratus, Physalopteroides venancioi, Physaloptera sp., an unidentified acuariid nematode and an unidentified centrorhynchid acanthocephalan. Except for Hexametra boddaertii (found only in Grussaí) and Pulchrosomoides elegans (found only in Praia das Neves), all helminth species were present at both localities. Half of the helminth species were present only as larvae and, in most cases, appear to represent paratenic parasitism. Overall prevalences of infection were high for both host species in both localities. Mabuya agilis tended to have richer and more diverse infracommunities than M. macrorhyncha. Some parameters of infection by individual helminth species seem to be related to the ecology of each Mabuya species. The parasite faunas were qualitatively very similar among species and/or localities, but quantitative similarities were more varied, due to differential representativeness of individual helminth species among host populations. The helminth communities of both skink species can be classified as non-interactive, being composed of site-specialists and immature stages of non-lizard parasites.

Studies were carried out to determine the predilection sites of Trichinella nativa muscle larvae in arctic foxes (Alopex lagopus) caught in Greenland. The highest number of larvae per gram of tissue was found in the muscles of the eyes and the legs. With regard to predilection sites no significant differences were demonstrated either between age groups or between foxes with high and low total parasite burdens. Predilection sites were comparable with those recorded earlier in experimentally infected caged foxes and in other carnivorous species. Hypotheses on predilection sites of Trichinella muscle larvae are discussed.

The interaction between Duddingtonia flagrans and infective larvae of Haemonchus contortus was studied in vitro under optical and scanning electron microscopy. Trap formation by the fungus started 9 hours after inoculation and first larvae were found 11 hours after larval inoculation on colonies grown on the surface of dialysis membranes. Scanning electron micrographs were taken 12, 24, 36 and 48 h after larval predation. Details of predation structures and fungus–larvae interaction are described. A mucilaginous substance occurred at the points of adherence of traps to nematode cuticle. Bacteria were also found at some points of interaction between fungus and larval cuticle. Cuticle penetration by fungus hyphae occurred only 48 h after predation.