The potential impact of exotic Lymnaea spp. on fascioliasis in Australasia

The presence of the following exotic Lymnaea spp. has been recently confirmed in Australia: L. columella (North and South America, South Africa, New Zealand,) L. viridis (China, Japan, Philippines, New Guinea, Pacific Area), L. auricularia rubiginosa (Indochina, Malaysia, Indonesia) and L. peregra (Europe).In the present studies the relative susceptibility of the exotic snails noted above to Fasciola hepatica was investigated. Two of the species, L. columella and L. viridis, were found to be highly susceptible to the Australian strain of F. hepatica and a substantial number of viable metacercariae were produced. However a strong disparity in the host— parasite relationship was evident when the parthenogenetic development of the fluke in the two exotic snails was compared with that in the native L. tomentosa. It was concluded that L. tomentosa is a more suitable intermediate host for F. hepatica than any of the introduced exotic species, but it has to be considered that the host—parasite relationships between new combinations of Fasciola and Lymnaea spp. may improve by adaptation. In view of the results of the present studies the potential danger of fascioliasis spreading to areas currently not endemic in Australasia through introduced exotic snails is discussed with particular reference to Queensland, Western Australia and the Northern Territory.There is strong circumstantial evidence that the exotic snails were introduced with aquatic plants through the relatively free overseas and Australian trade. Similarly, through the introduction of infected snails, dangerous parasites such as F. gigantica of cattle and sheep or Schistosoma ssp. which cause serious diseases of man and animals could enter the subtropical or tropical regions of Australia. Appropriate State and Federal quarantine regulations are essential to prevent further introduction and spread of the snails, and monitoring of the distribution of established exotic snails in Australasia must be continuous.     

Prevalence of bovine fascioliasis in a new-emerging focus of human fascioliasis in BoyerAhmad district,southwest of Iran

BackgroundThe prevalence of fascioliasis in a population of cattle in a new-emerging focus of human fascioliasis in Boyer-Ahmad District, in the southwest of Iran, was investigated, using an ELISA system and coprological methods.MethodsBlood, as well as stool samples, were collected from 150 dairy cattle consisted of 82 (54.7%) males and 68 (45.3%) females, aged from 1 to 8 years, selected from different pastures. The stool samples were evaluated by direct wet mount microscopic examination as well as sedimentation methods. Moreover, modified Telemann was used for detecting of Fasciola eggs. Anti-Fasciola antibodies in the cattle sera samples were determined, using a Fasciola excretory-secretory antigen-specific ELISA.ResultsAnti-Fasciola antibodies were detected in 64 (42.4%) of the subjects by ELISA. Out of 64 seropositive cases, 29 (45.3%) were female and 35 (54.6%) were male. No significant association was found between sex and seropositivity to fascioliasis. Considering the age of the cattle, the highest prevalence of fascioliasis (23%) was found in the 5-year old age group. However, the difference between the age of the cattle and seropositivity to fascioliasis was not statistically significant (P > 0.05). Fasciola eggs were not detected in fecal samples of any of the studied cattle, while Paramphistomide, as well as Dicrocoelium dendriticum eggs, were detected in 6 (4%) and 17 (11.3%) of the subjects, respectively.ConclusionFindings of the current study provide basic information about bovine fascioliasis in a new focus of human fascioliasis in Iran, which is needed for effective control of this parasitic infection in such areas.     

Prevalence of Fasciola in cattle and of its intermediate host Lymnaea snails in central Vietnam

The aims of this study were to investigate the prevalence of natural Fasciola infections in both the definitive hosts (cattle) and the intermediate hosts (Lymnaea snails) in central Vietnam. A total of 1,075 fecal samples, randomly collected from cattle in Binh Dinh, Khanh Hoa, and Phu Yen provinces, were examined for Fasciola eggs by a sedimentation method. The overall prevalence of Fasciola was 45.3?%. A subset of the animals (235) was also screened for antibodies against Fasciola by an enzyme-linked immunosorbent assay. Overall, 46.3?% of these animals were shedding Fasciola eggs while 87.2?% were Fasciola seropositive. A lower prevalence of Fasciola was observed in calves ??2?years of age (37.6?%) compared to that in cattle >2?years of age (53.7?%) (p?<?0.05). The prevalence in the rainy season (50.8?%) was significantly different to that in the dry season (38.1?%) (p?<?0.05). Of the 3.269 Lymnaea viridis and 1.128 Lymnaea swinhoei examined, 31 (0.95?%) and seven (0.62?%), respectively, were found to be infected with Fasciola. This appears to be the first epidemiological survey of the prevalence of Fasciola in cattle and snails in these three provinces in central Vietnam.     

A comparison of two laboratory methods of maintaining field-collected Lymnaea tomentosa for the production of Fasciola hepatica metacercariae

Four replicates of 50 field-collected Lymnaea tomentosa were maintained in two laboratory culture systems and their efficiency and practicability for the production of Fasciola hepatica metacercariae were assessed. There was no significant difference in the total number of metacercariae produced (56 552 from 64 snails in the closed container system and 54 073 from 82 snails in the shallow aquarium system).There was no significant difference between culture systems as assessed by snail survival, the number of infected snails or the number of infected snails harvested for the recovery of metacercariae. Within both systems there was a significant effect of snail size on the number of metacercariae produced. Optimum survival and production of metacercariae was achieved in young adult snails of 5–8 mm shell length at the time of infection.The collection of L. tomentosa from the field habitat and their maintenance in either culture system was shown to be a practical alternative to the breeding of laboratory adapted strains of L. tomentosa for the regular production of large numbers of F. hepatica metacercariae.     

MM3-ELISA evaluation of coproantigen release and serum antibody production in sheep experimentally infected with Fasciola hepatica and F. gigantica

During an experimental infection of sheep with Fasciola hepatica or F. gigantica, MM3-SERO and MM3-COPRO ELISA tests were applied to compare the kinetics of antibody production and coproantigen release between the 2nd and 32nd week post-infection (wpi). The Kato-Katz technique was used to measure the kinetics of egg shedding by both Fasciola species (eggs per gram of feces, epg). The kinetics of IgG antibodies for all sheep infected with F. hepatica and F. gigantica followed a similar pattern. Optical density (OD) increased rapidly between the 4th until the 12th wpi, when the highest values were reached and then decreased slowly until the 32nd wpi. Coproantigen levels increased above the cut-off value between 6 and 9 wpi in the F. hepatica group, and between 9 and 11 wpi in the F. gigantica group. The comparison between coproantigen levels and epg indicated that F. hepatica-infected sheep had detectable amounts of coproantigens 4–7 weeks before patency (egg shedding), while F. gigantica-infected sheep had detectable amounts of coproantigens 3–6 weeks before patency. When comparing the kinetics of coproantigen release vs the kinetics of epg, a similar pattern emerged, but with a two-week time-lag in epg, for both F. hepatica and F. gigantica infections. The amount of coproantigen release by each adult was not burden dependent for F. hepatica infection (burden of 33–66 adults), while it was for F. gigantica infection (burden of 17–69 adults). The results demonstrate the usefulness of the MM3-SERO and MM3-COPRO ELISAs as tools for the diagnosis of early as well as long-term fascioliasis infections, and suggest that they can potentially be applied to human fascioliasis even in countries where F. hepatica and F. gigantica co-exist. These tests can be employed not only in the diagnosis, but also in studies on epidemiology as well as pathogenesis and treatment in animals and humans since they allow post-treatment infection monitoring.     

Human and animal fasciolosis: Coprological survey in Narok,Baringo and Kisumu counties,Kenya

Fasciolosis is caused by digenean trematodes of the genus Fasciola. The principal definitive hosts are cattle, sheep and goats. Humans are infected as accidental hosts. Fasciolosis is one of the major neglected tropical diseases and is considered an emerging zoonotic infection. This study was aimed at determining the prevalence of human and domestic animal fasciolosis in selected counties in Kenya. Stool samples for Fasciola diagnosis were collected from humans and domestic animals and transported to the laboratory at Egerton University and processed using sedimentation technique and examined for the presence of eggs. A total of 272 human samples collected were all negative for Fasciola eggs. A total of 582 domestic animals (cattle [46.0%], sheep [29.9%] and goats [24.1%]) samples collected had overall prevalence of 30.9% for Fasciola infection. There was no significant differences (p > 0.05) between the prevalence of fasciolosis and origin of the animals, sex and season. There was a significant difference (p < 0.05) between the prevalence of fasciolosis and domestic animals, age and body condition. The prevalence of fasciolosis was high in two irrigation schemes which favour the breeding of intermediate host snail and grazing of animals along the irrigation canals where metacercaria of Fasciola parasites could be present on the vegetation. Although human fasciolosis was not detected in this study, the presence of animal fasciolosis can pose public health risk because of its zoonotic nature. Therefore, it is important to introduce measures which would help to reduce the exposure of animals to Fasciola infection.     

Experimental Infection of Lymnaea palustris snails with Fascioloides magna

This report describes experimental infection of Lymnaea (Galba) palustris. (Müller, 1774) snails with Fascioloides magna (Bassi, 1875). Of 725 L. palustris, 400 survived the experimental infection and in 172 (43%) the developmental cycle of the fluke was completed. Free emergence of cercariae was obtained was obtained 68 days after infection. The infectivity of metacercariae thus obtained was demonstrated in guinea-pigs, rabbits and a Cameroon goat. In sheep, the infection produced only changes in the organs, but no flukes could be recovered. Considering that L. palustris are found frequently in natural habitats in Czechoslovakia, they should be considered as possible intermediate hosts for F. magna.     

Growth response in lambs injected with a long acting microencapsulated Vitamin B 12

Nematode parasites have been a major factor limiting sheep production in New Zealand for more than 100 years. Twenty-nine species of nematodes were unintentionally introduced with sheep into New Zealand, but it is principally species of Haemonchus, Ostertagia, Trichostrongylus, Nematodirus and Cooperia that are associated with production losses and clinical disease.

The seasonal dynamics of nematode infection are the consequence of complex inter-relationships between the sheep, their husbandry and the prevailing climate. The patterns of pasture contamination by nematode eggs and then larvae, and the subsequent levels of infection in ewes and lambs are broadly similar throughout New Zealand. Numbers of infective larvae on pasture build up over summer to a peak in autumn/early winter with, in some years, a spring peak derived from the parturient rise in faecal nematode egg counts (FEC), expressed in eggs per gram of faeces (epg), in lactating ewes. The immune capability of lambs is initially low but increases with the magnitude and duration of exposure to infection. Once significant immunity has developed (usually by 10–12 months of age), sheep are capable of markedly restricting parasite infection, except during times of disease, malnutrition or stress.

For the effective control of nematode parasites, farmers have come to rely almost exclusively on broad-spectrum anthelmintics. However, issues relating to resistance, residues and eco-toxicity increasingly threaten the sustainability of chemotherapy. In order to maintain present levels of parasite control and productivity in the long term, farmers need to integrate management practices aimed at minimising animal exposure to parasites with reduced reliance on anthelmintics.     

Environmental determinants of the spatial distribution of Angiostrongylus vasorum,Crenosoma vulpis and Eucoleus aerophilus in Hungary

Angiostrongylus vasorum, Crenosoma vulpis and Eucoleus aerophilus (syn. Capillaria aerophila) are the most important lungworm species infecting wild and domesticated canids in Europe. To investigate the spatial distribution of these parasites and the factors influencing their circulation in the fox populations, 937 red foxes (Vulpes vulpes) were tested for lungworm infection in Hungary. The prevalence of A. vasorum, C. vulpis and E. aerophilus infection was high (17.9, 24.6 and 61.7%). The distribution pattern of infection in foxes and the relationship of this pattern with landscape and climate was analyzed by geographic information system. Based on the analysis, the annual precipitation was the major determinant of the spatial distribution of A. vasorum and C. vulpis and E. aerophilus. Nevertheless, the mean annual temperature also influenced the distribution of A. vasorum and E. aerophilus. The positive relationship with annual precipitation and the negative relationship with mean annual temperature can be attributed to the sensitivity of larvae, eggs and intermediate hosts (snails and slugs) of lungworms for desiccation. Based on the highly clumped distribution of A. vasorum and C. vulpis, the indirect life cycle (larvae, slugs and snails) of these parasites seems to be particularly sensitive for environmental effects. The distribution of E. aerophilus was considerably less clumped indicating a lower sensitivity of the direct life cycle (eggs) of this parasite for environmental factors. Based on these results, lungworm infections in canids including dogs can be expected mainly in relatively wet and cool areas.     

An epidemiological study of Fasciola hepatica in the Netherlands

Summary

Transmission of F.hepatica under natural conditions was analysed in a three year programme. The variables used were the indirect haemagglutination (IHA) technique, worm establishment in tracer lambs and the population dynamics, infection rate and shedding pattern of Lymnaea truncatula.

It is concluded that fluke eggs, infected snails and metacercariae on herbage can survive the winter in the Netherlands. Metacercarial availability was positively correlated to the amount of rainfall in the grazing period. The role of developed eggs that survive the winter is important, because this results in earlier infections in the herd.

The use of the serological diagnosis method IHA is important to detect F. hepatica infection in an early stage. Use of cellophane paper on floats is a useful method for determining the shedding pattern of cercariae from L. truncatula. It is concluded that collection of metacercariae on cellophane floats, inventarization of L. truncatula and its infection level are useful tools for the prediction of liverfluke infections.     

Identification of an isolate of Saprolegnia ferax as the causal agent of saprolegniosis of Yellow catfish (Pelteobagrus fulvidraco) eggs

Saprolegnia species have been implicated for significant fungal infections of both living and dead fish as well as their eggs. In the present study, an oomycete water mould (strain HP) isolated from yellow catfish (Peleobagrus fulvidraco) eggs suffering from saprolegniosis was characterized both morphologically and from ITS sequence data. It was initially identified as a Saprolegnia sp. isolate based on its morphological features. The constructed phylogenetic tree using neighbour joining method indicated that the HP strain was closely related to Saprolegnia ferax strain Arg4S (GenBank accession no. GQ119935), that had previously been isolated from farming water samples in Argentina. In addition, the zoospore numbers of strain HP were markedly influenced by a variety of environmental variables including temperature, pH, formalin and dithiocyano-methane. Its zoospore formation was optimal at 20?°C and pH?7, could be well inhibited by formalin and dithiocyano-methane above 5?mg/L and 0.25?mg/L, respectively. To our knowledge, this is the first report on the S. ferax infection in the hatching yellow catfish eggs.     

Epidemiology of fascioliasis in the koshi hills of Nepal

Summary Lymnaea auriculariarace rufescensand Lymnaea luteolawere shown to transmit F. giganticain the Koshi hills of Nepal. The ecology of the snails was studied at nine different habitats in Hattikharka panchayat, the highest numbers occurring immediately after the end of the monsoon. Mature F. giganticainfections in the snails were detected from May to August and again in November. The prevalence of the infection in cattle was highest during the summer monsoon and in January and February. A control programme for fascioliasis based on these findings is suggested which requires that cattle be treated in February and possibly again in late August.

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Epidemiologia De Fascioliasis En Las Colinas Koshi De Nepal

Resumen Se encontró que laLymnaea auricularia razarufescens y laLymnea luteola transmitenF. gigantica en las colinas Koshi de Nepal. Se estudió la ecología de los caracoles en 9 habitats diferentes en Hattikharka panchayat, ocurriendo el mayor número inmediatamente despues de los monsones. Las infecciones de las caracolas con fases maduras deF. gigantica, se detectaron desde mayo hasta agosto y otra vez en noviembre. La prevalencia de la infección en bovinos fue más alta durante los monsones veraneros, y posteriormente en enero y febrero. Se sugiere el tratamiento del ganado en febrero y agosto para controlar la fascioliasis.

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Epidemiologie De La Fasciolose Dans Les Collines De Koshi Au Nepal

Résumé On a montré queLymnea auricularia racerufescens etLymnea luteola transmettentF. gigantica dans les collines de Koshi, au Népal. L'écologie des mollusques a été étudiée dans 9 habitats différents du panchayat d'Hatikharka; le plus grand nombre se rencontre immédiatement après la fin de la mousson. Des infections àF. gigantica matures ches les escargots ont été détectées de mai à août, et de nouveau en novembre. La prévalence de l'infestation chez le bétail est maximum pendant la mousson d'été, puis en janvier et février. Un programme de contrôle de la fasciolose est suggéré sur la base de ces découvertes; il requiert que le bétail soit traité en février et, si possible, de nouveau fin août.

     

Serodiagnosis of Metastrongylus apri infection of pigs by immunofluorescence

The lyophilised first stage larvae of Metastrongylus apri were used as antigen in the indirect fluorescent antibody test for the diagnosis of the infection of pigs. The larvae were recovered from the embryonated eggs discharged by the gravid females of the nematode in vitro. Ten pigs were used in the test. Serial serum samples were taken of seven pigs, experimentally infected with varying doses of M. apri larvae, all of which were found to discharge eggs of the nematode in their faeces after infection was established. The other three pigs were uninfected and used as controls.Positive cuticular fluorescence of the larvae was first detected with the serum samples obtained between 14 and 33 days post-infection. This fluorescence persisted with subsequent serum samples up to 85 days post-infection. Pronounced and uniform cuticular fluorescence was generally observed with the serum dilutions of up to $\text{1}\text{10}$. Earlier post-infection serum samples either exhibited no cuticular fluorescence or gave non-brilliant fluorescence. The pre-infection serum of these pigs and also serum samples obtained from the three uninfected control pigs did not give cuticular fluorescence even at the initial serum dilution of $\text{1}\text{5}$.     

Dynamics of the free-living stages of sheep intestinal parasites on pasture in the North Island of New Zealand. 2. Weather variables associated with development

Abstract

AIM: To identify weather variables associated with the development of eggs of Teladorsagia (=Ostertagia) circumcincta, Trichostrongylus colubriformis and Haemonchus contortus to third-stage infective larvae (L3) under a range of climatic conditions on pasture in the North Island of New Zealand.

METHODS: Sheep faeces containing known numbers of eggs of all three nematode species were deposited on, or buried in, pasture plots at three sites, viz coastal Manawatu, Upper Hutt Valley, and East Cape hill country. Development was measured by recovering L3 from faeces, herbage and soil 28–31 days after deposition, on 13–18 occasions between January 2005 until July 2006. Weather data were recorded at each site, and the association between weather variables and number of L3 recovered was analysed using subsets regression to select best-fitting models from several candidate variables, after adjustment for efficiency of recovery of L3. A multiple linear regression model was then developed for each species, to select weather variables that had both significant and substantive effects on the number of L3 recovered.

RESULTS: For all species, mean daily temperature was the bestpredictor of the number of L3 recovered (p=0.001). For T.circumcincta the final model included mean daily temperature and soil temperature (R²=51%), and for T. colubriformis the model only included mean daily temperature (R²=55%). For development of H. contortus, mean daily temperature was the most significant variable, but moisture in the form of rainfall entropy, i.e. the temporal distribution of rainfall, overthe first 14 days was also significant in the final model (R²=34%).

CONCLUSIONS: Temperature was the most important determinant of developmental success of free-living nematodes on pasture at the study sites, and probably also for other parts ofNew Zealand with similar climates. Moisture was not significant in the development of T. circumcincta or T.colubriformis, implying that under the generally moist temperate climate in New Zealand, moisture is seldom limiting for development of these species. Haemonchus contortusappeared to be sensitive to moisture availability and precipitation in the first 14 days after deposition of faeces. The results of this study will be further developed as part of species-specific climate-driven models of parasite development in New Zealand.     

Epidemiological studies of Fasciola gigantica infections in cattle in the highveld and lowveld communal grazing areas of Zimbabwe

During the period between January 1999 and December 2000, the distribution and seasonal patterns of Fasciola gigantica infections in cattle in the highveld and lowveld communal grazing areas of Zimbabwe were determined through monthly coprological examination. Cattle faecal samples were collected from 12 and nine dipping sites in the highveld and lowveld communal grazing areas respectively. Patterns of distribution and seasonal fluctuations of the intermediate host-snail populations and the climatic factors influencing the distribution were also determined by sampling at monthly intervals for a period of 24 months (November 1998 to October 2000) in six dams and six streams in the highveld and in nine dams in the lowveld communal grazing areas. Each site was sampled for relative snail density and the vegetation cover and type, physical and chemical properties of water, and mean monthly rainfall and temperature were recorded. Aquatic vegetation and grass samples 0-1 m from the edges of the snail habitats were collected monthly to determine the presence or absence of F. gigantica metacercariae. Snails collected at the same time were individually checked for the emergence of larval stages of F. gigantica. A total of 16264 (calves 5418; weaners 5461 and adults 5385) faecal samples were collected during the entire period of the study and 2500 (15.4%) of the samples were positive for F. gigantica eggs. Significantly higher prevalences were found in the highveld compared to the lowveld (P < 0.001), for adult cattle than calves (P < 0.01) and in the wet season over the dry season (P < 0.01). Faecal egg output peaked from August/September to March/April for both years of the study. Lymnaea natalensis, the snail intermediate host of F. gigantica was recorded from the study sites with the highveld having a significantly higher abundance of the snail species than the lowveld (P < 0.01). The snail population was low between December and March and started to increase in April reaching a peak in September/October. The number of juvenile snails peaked between April and August. The mean number of snails collected was negatively correlated with rainfall and positively correlated with temperature. Mean number of snails collected was also positively correlated with Potamogeton plant species and negatively correlated with Cyperus plant species. However, none of the L. natalensis collected from the habitats were found shedding Fasciola cercariae. Metacercariae were found on herbage from the fringes of the snail habitats between February and August for both years, with most of the metacercariae concentrated on herbage 0-1 m from the banks of the habitats. Based on the findings of this study, anthelmintic treatment should be administered in December/January to control chronic and mature fasciolosis. A second treatment should be given in April/May to reduce pasture contamination and subsequently snail infection, as this is the time the snail population starts to build up. To control acute fasciolosis due to the immature liver flukes a third treatment should be given in August. The first application of molluscicides to control the snail intermediate hosts can be done in June the time when the snail is harbouring the parasite and a second application in September in order to kill new generations of infected snails     

Development of immune responsiveness to Ascarissuum antigens in pigs vaccinated with ultraviolet-attenuated eggs

Pigs fed $\text{Ascaris}$$\text{suum}$ eggs attenuated by short wave ultraviolet-radiation developed resistance to challenge infection. Per os inoculation of pigs on three successive weeks with 10,000 eggs irradiated to total exposures of 150,100 and 75 μW-min/cm², respectively, resulted in an 88% reduction in the number of larvae recovered from the lungs 7 days after challenge with 10,000 infective eggs. Peripheral blood lymphocytes (PBL) from vaccinated pigs were specifically stimulated in vitro to incorporate tritiated thymidine by egg hatching fluid (Ea) and by excretory-secretory products obtained from cultures in defined-media of second-stage larvae (L2) developing to third-stage (L2–3) and from cultures of third-stage larvae developing to fourth-stage (L3–4). PBL were also specifically stimulated by living L2. Ea and L2 stimulated pig PBL significantly at 7 days after the first inoculation; responses to L2–3 and L3–4 developed 7 days after a second inoculation. The antigen-responsive cells in the PBL population were non-immunoglobulin-bearing lymphocytes. Antibodies to Ea and L2–3 were not detected in the serum of vaccinated pigs, and only 3 of 7 pigs had low concentrations of serum antibodies to L3–4.     

Effects of Three Anthelmintic Treatment Regimes against Fasciola and Nematodes on the Performance of Ewes and Lambs on Pasture in the Highlands of Kenya

The efficacy of strategic anthelmintic control of liver flukes (Fasciola) and gastrointestinal (GI) nematodes on the performance of ewes and lambs on pasture was assessed on a farm in the highlands of Kenya. In May 1999, 45 Corriedale ewes, aged between 2 and 3 years, were ear-tagged, weighed and allocated randomly to three equal treatment groups based on body weight. Faecal samples taken at this time revealed low levels of strongyle-type eggs per gram of faeces (EPG) and the presence of liver fluke eggs in only a few of the animals. All the animals were then set stocked for 12 months on separate paddocks in an area endemic for both Fasciola and GI nematodes. The ewes in group 1 were given a combined anthelmintic treatment against Fasciola and GI nematodes during the periods recommended for the control of Fasciola in the area (February, June and October). The ewes in group 2 were given the combined treatments 3 weeks after the onset of both the short and long rainy seasons (November and April, respectively). Those in group 3 were given separate treatments for Fasciola (February, June and October) and nematodes (3 weeks after the onset of the rainy seasons). The anthelmintic treatment against Fasciola consisted of triclabendazole at 10 mg per kg body weight, and that against nematodes was levamisole at 10 mg per kg body weight.The nematode EPG for the ewes in group 1 were higher than in groups 2 and 3 during both rainy seasons. The nematode EPG did not differ significantly between groups 2 and 3. The prevalence of Fasciola eggs (number of ewes shedding eggs in a group) in the ewes in groups 1 and 3 remained very low throughout the study period compared to those in group 2. The highest birth weights and the weight gains of lambs were recorded for the group of ewes given separate anthelmintic treatments for Fasciola and nematodes (group 3). The results of this trial indicated that, in an area like Nyandarua District, where liver flukes and GI nematodes are important constraints to sheep production, the best practice is to give separate treatments for the two groups of parasites at the recommended times.     

Strongyle infections of small ruminants in Nigeria

A survey of strongyle infections was conducted in sheep and goats reared in a traditional e extensive husbandry system in two ecological zones if Nigeria. One zone had a seasonal pattern of infection. The majority of animals had faecal worm parasite egg counts of below 500 eggs per gram. Kids, and lambs younger than 3 months did not carry strongyle worm burdens, and the highest infection rate was found in the 7–12 month age group. A high proportion of small ruminants shed strongyle eggs during the postparturient period.The helminth species found by the use of larval culture techniques on the faeces were: Haemonchus contortus, Trichostrongylus colubriformis and Oesophagostonum columbianum. Adults of the same species were found in the few animals necropsied. The significance of the findings is discussed.     

New multiplex PCR method for the simultaneous diagnosis of the three known species of equine tapeworm

Although several techniques exist for the detection of equine tapeworms in serum and feces, the differential diagnosis of tapeworm infection is usually based on postmortem findings and the morphological identification of eggs in feces. In this study, a multiplex polymerase chain reaction (PCR)-based method for the simultaneuos detection of Anoplocephala magna, Anoplocephala perfoliata and Anoplocephaloides mamillana has been developed and validated. The method simultaneously amplifies hypervariable SSUrRNA gene regions in the three tapeworm species in a single reaction using three pairs of primers, which exclusively amplify the internal transcribed spacer 2 (ITS-2) in each target gene. The method was tested on three types of sample: (a) 1/10, 1/100, 1/500, 1/1000, 1/2000 and 1/5000 dilutions of 70 ng of genomic DNA of the three tapeworm species, (b) DNA extracted from negative aliquots of sediments of negative control fecal samples spiked with 500, 200, 100, 50 and 10 eggs (only for A. magna and A. perfoliata; no A. mamillana eggs available) and (c) DNA extracted from 80, 50, 40, 30, 10 and 1 egg per 2 μl of PCR reaction mix (only for A. magna and A. perfoliata; no A. mamillana eggs available). No amplification was observed against the DNA of Gasterophilus intestinalis, Parascaris equorum and Strongylus vulgaris. The multiplex PCR method emerged as specific for the three tapeworms and was able to identify as few as 50 eggs per fecal sample and as little as 0.7 ng of control genomic DNA obtained from the three species. The method proposed is able to differentiate infections caused by the two most frequent species A. magna or A. perfoliata when the eggs are present in feces and is also able to detect mixed infections by the three cestode species.     

The laboratory maintenance of field-collected Lymnaea tomentosa for the production of Fasciola hepatica metacercariae

A culture method is described for the production of Fasciola hepatica metacercariae in Lymnaea tomentosa collected from the field habitat. Two-litre glass jars, filled with artificial spring (A.S.) water and having a substratum of soil-bentonite (S.B.) agar to provide habitat and food, maintain groups of 40 to 100 snails graded according to size. Small quantities of washed lettuce are fed as a supplement. Compressed air, pre-warmed to 25°C, is piped to each jar to maintain thermoregulation, aeration, filtration and circulation of the water. Infection is achieved by reducing the A.S. water level to 3–4 mm and exposing the snails to miracidia for 6 h. During the aquatic developmental phase of 5–6 weeks, twice-weekly changes of A.S. water and the filter minimise proliferation of protozoans and Chaetogaster spp. Accumulation of mature cercariae is achieved by sustaining the infected snails on moist S.B. agar while humidification within the jar is maintained. Alternatively, groups of five to ten snails may be held in disposable petri dishes lined with S.B. agar. Maturation of cercariae may be delayed by refrigeration at 4–5°C. Upon maximum accumulation, the cercariae are harvested by dissection of the snails and metacercariae are stored in plastic containers at 4°C.