Zoonotic transmission of Teladorsagia circumcincta and Trichostrongylus species in Guilan province, northern Iran: molecular and morphological characterizations.

BACKGROUND: Parasitic trichostrongyloid nematodes have a worldwide distribution in ruminants and frequently have been reported from humans in Middle and Far East, particularly in rural communities with poor personal hygiene and close cohabitation with herbivorous animals. Different species of the genus Trichostrongylus are the most common trichostrongyloids in humans in endemic areas. Also, Ostertagia species are gastrointestinal nematodes that mainly infect cattle, sheep and goats and in rare occasion humans. The aim of the present study was to identify the trichostrongyloid nematodes obtained from a familial infection in Guilan province, northern Iran, using morphological and molecular criteria. METHODS: After anthelmintic treatment, all fecal materials of the patients were collected up to 48 h and male adult worms were isolated. Morphological identification of the adult worms was performed using valid nematode keys. Genomic DNA was extracted from one male worm of each species. PCR amplification of ITS2-rDNA region was carried out, and products were sequenced. Phylogenetic analysis of the nucleotide sequence data was performed using MEGA 6.0 software. RESULTS: Adult worms expelled from the patients were identified as T. colubriformis, T. vitrinus and Teladorsagia circumcincta based on morphological characteristics of the males. Phylogenetic analysis illustrated that each species obtained in current study was placed together with reference sequences submitted to GenBank database. CONCLUSIONS: The finding of current study confirms the zoonotic aspect of Trichostrongylus species and T. circumcincta in inhabitants of Guilan province. The occurrence of natural human infection by T. circumcincta is reported for the first time in Iran and the second time in the world.

Host immunity shapes the impact of climate changes on the dynamics of parasite infections.

Global climate change is predicted to alter the distribution and dynamics of soil-transmitted helminth infections, and yet host immunity can also influence the impact of warming on host-parasite interactions and mitigate the long-term effects. We used time-series data from two helminth species of a natural herbivore and investigated the contribution of climate change and immunity on the long-term and seasonal dynamics of infection. We provide evidence that climate warming increases the availability of infective stages of both helminth species and the proportional increase in the intensity of infection for the helminth not regulated by immunity. In contrast, there is no significant long-term positive trend in the intensity for the immune-controlled helminth, as immunity reduces the net outcome of climate on parasite dynamics. Even so, hosts experienced higher infections of this helminth at an earlier age during critical months in the warmer years. Immunity can alleviate the expected long-term effect of climate on parasite infections but can also shift the seasonal peak of infection toward the younger individuals.

Recovery of strongylid third-stage larvae from herbage samples: standardisation of a laboratory method and its application in the field.

The estimation of parasitic nematode larvae present on pasture is an important tool applied in many epidemiological studies. In the face of climatic changes, there is increased interest in identifying parameters influencing the survival of free-living stages of parasites under different meteorological conditions. In order to predict possible risk factors for grazing livestock, reliable and reproducible methods to assess the density of larvae on pasture are required. A laboratory method for the recovery of strongylid third-stage larvae from herbage samples was developed, standardised and its efficacy assessed in controlled experiments as well as under field conditions. Grass samples free of any nematode larvae were used and inoculated with known numbers of third-stage larvae of Cooperia oncophora in different concentrations. The grass samples were inoculated with larvae over 24 h, followed by soaking for 4 h. The recovery process included washing over sieves and centrifugation of the obtained liquid. The total time required for the recovery process was about 5-7 h (excluding inoculation). Recovery rates range from 68% to 98% and a strong correlation between numbers of larvae added to the grass samples and numbers of larvae that could be recovered was observed (p < 0.001). The new method proved to be reproducible and provides high recovery rates combined with the potential to handle many samples simultaneously in a relatively short time, thus offering high throughput opportunities applicable to field experiments.

The influence of water on the migration of infective trichostrongyloid larvae onto grass.

Detailed knowledge of the effects of water on the migration of infective larvae of economically important trichostrongyloid species is urgently needed to feed into prediction models of future epidemiology. The influence of water on the migration of the parasitic nematodes Nematodirus battus, Haemonchus contortus and Teladorsagia circumcincta from sheep dung onto grass was examined in a series of laboratory experiments. Turf plots were seeded with larvae, which were recovered from grass clippings by serial sampling. Free water was necessary for larvae to escape from dung, but not for vertical migration onto grass. When temperature and relative humidity were held constant, the proportion of a population of live larvae present on herbage reached a plateau of around 2 (1-10)% after 24 h, and then changed little over time. Larvae in soil and dung formed a reservoir, such that a similar proportion of the larval population was maintained on grass after clipping. These findings suggest continuing random movement of free larvae. Implications for the epidemiology of trichostrongyloid species are discussed in the context of trade-offs faced by the parasites.

Adaptations and phenotypic plasticity in developmental traits of Marshallagia marshalli.

Despite the economic, social and ecological importance of the ostertagiine abomasal nematode Marshallagia marshalli, little is known about its life history traits and its adaptations to cope with environmental extremes. Conserved species-specific traits can act as exaptations that may enhance parasite fitness in changing environments. Using a series of experiments, we revealed several unique adaptations of the free-living stages of M. marshalli that differ from other ostertagiines. Eggs were isolated from the feces of bighorn sheep (Ovis canadensis) from the Canadian Rocky Mountains and were cultured at different temperatures and with different media. Hatching occurred primarily as L1s in an advanced stage of development, morphologically very similar to a L2. When cultured at 20 °C, however, 2.86% of eggs hatched as L3, with this phenomenon being significantly more common at higher temperatures, peaking at 30 °C with 28.95% of eggs hatching as L3s. After hatching, free-living larvae of M. marshalli did not feed nor grow as they matured from L1 to infective L3. These life history traits seem to be adaptations to cope with the extreme environmental conditions that Marshallagia faces across its extensive latitudinal distribution in North America and Eurasia. In order to refine the predictions of parasite dynamics under scenarios of a changing climate, basic life history traits and temperature-dependent phenotypic behaviour should be incorporated into models for parasite biology.

[Biology of gastrointestinal nematodes of ruminants]. / Biologia dei nematodi gastrointestinali dei ruminanti.

The development and survival of free-living stages of gastro-intestinal nematodes of small ruminants are influenced by several abiotic and biotic factors. Within the abiotic factors, most important are the environmental temperature and humidity. They regulate the development of larvae from eggs dispersed on the pasture by the animals faeces. Each parasite species that infect ruminants requires a different time to development, depending on temperature and humidity. Among trichostrongylids, Ostertagia, Teladorsagia and Nematodirus show a strong adaptation to low temperatures. Nematodirus larvae are able to survive to winter inside the egg shell. Temperature and humidity influence the distribution and survival of larvae on pasture. The larval third stage can migrate from faeces to pasture vegetation and they accumulate at the basis of vegetation where stay during the day or in the soil to avoid the desiccation. The forage species affects the migration of larvae on herbage too. Many biological factors contribute to disperse the larvae on the pasture. Dung burying beetles, coprophagous beetles and earthworms can greatly reduce the larvae of some trichostrongylids on pasture. They contribute to the spread of the faecal material on the pasture and allow the larval death as a consequence of drying.

Very low helminth infection in sheep grazed on pastures fertilised by sewage sludge or cattle slurry.

Helminth infection acquired by lambs grazing on pastures fertilised either by urban sewage sludge or cattle slurry were studied in temperate Central Western France. The aim was to assess the risk of larval cestodoses in lambs after sewage application and of digestive tract nematode infection following the slurry application. Twenty-six sheep were allocated on two paddocks of 0.7 ha, one fertilised with sludge and the other with cattle slurry. The delay between application and actual grazing was 6 weeks; grazing on these paddocks extended from mid July to beginning of November 2002. The herbage biomass was slightly increased in the sludge paddock but it did not result in an increase of lamb live weight, compared with the slurry paddock. The lambs did not acquire cysticercosis or any other larval cestodoses in the sewage sludge group and only very limited infections with Cooperia spp. and Nematodirus spp. were observed in the slurry group. It was concluded that in our conditions the helminth risk was extremely low and was not a cause of restriction of the use of these biowastes.

The influence of weather and egg contamination on the development of third-stage larvae of Cooperia oncophora on pasture.

The influence of weather and egg contamination on the dynamics of herbage contamination with infective larave of Cooperia oncophora was investigated on artificially contaminated grass plots and in a grazing experiment with 24 first-year grazing calves from May to October 1987 in Lower Saxony, Germany. On the experimental plots the larval translation was highest at the beginning of July and in the second part of September, following high mean weekly temperatures. Between July and September peak recovery of larvae from herbage occurred 4 weeks after contamination. A seasonal pattern of larval translation similar to that on the experimental plots could be demonstrated on the grazed pastures when the number of larvae per m2 of pasture had been adjusted to the previous egg output by means of a contamination index. The resulting 'relative larval density' is regarded as a good indicator for larval development on pasture. From July to September the larval population on pasture resulted mainly from the egg contamination 2-3 weeks earlier. The short persistence of the infective larvae on herbage was probably due to the frequent and heavy rainfall throughout the season, causing a passive washout of larvae into the soil. On single pastures the larval density started to increase within 1 week after the calves had first contact with these fields. The impact of the calves on the distribution of larvae is discussed.

Transmission dynamics of helminth parasites of pigs on continuous pasture: Oesophagostomum dentatum and Hyostrongylus rubidus.

An increase in alternative outdoor pig production systems is occurring in Denmark, and this study was designed to elucidate the transmission patterns of Oesophagostomum dentatum and Hyostrongylus rubidus in pigs allowed to graze continuously on a pasture. A group of pigs was turned out in May 1993 (Year 1 of the study) and subsequently inoculated with low numbers of both helminths. These pigs were followed parasitologically until October by serial necropsy and sampling of faeces, grass and soil. A non-inoculated group of pigs was similarly followed on the same pasture in Year 2 (1994). Pasture infectivity was measured using helminth-naïve tracer pigs during all seasons. The pasture vegetation was rapidly destroyed by the pigs, resulting in a dirt lot by the autumn of Year 2. The area was soon contaminated with eggs, resulting in heavy pasture infectivity and increasing worm burdens in late summer; then the numbers of larvae declined markedly. In May of Year 2, newly exposed pigs became only lightly infected (mostly O. dentatum), and no transmission was observed in July-August of Year 2, probably due to an unusually dry summer and a lack of protecting vegetation. The results indicate that both O. dentatum and H. rubidus are very sensitive to environmental factors, because significant transmission occurred only under the most favourable conditions (summer combined with protecting vegetation as in Year 1). Transmission was severely reduced during the low temperatures experienced in the winter between Years 1 and 2 and during the dry summer of Year 2, when vegetation was lacking. Continuous grazing actually reduced transmission of O. dentatum and H. rubidus because of the reduction in vegetation. This, however, is not a desirable alternative farming system, because of its adverse environmental effects. This environmental impact may be mitigated by employment of a pasture rotation system in place of continuous grazing.

Estimation of uptake of digestive tract strongyle larvae from pasture, using oesophagus fistulated sheep.

Three Merino of Arles ewes fistulated at the oesophagus were used in autumn and spring in order to assess their larval uptake on infected irrigated pasture. The rate of transmission (ingested/available larvae of digestive tract strongyle) was low when the pasture was heavily infected; it was not otherwise modified by environmental factors.

Limited role of lagomorphs (Oryctolagus cuniculus and Lepus capensis) in the dispersion of parasite nematodes of ruminants.

Rabbits (Oryctolagus cuniculus) and hares (Lepus capensis) were investigated under natural conditions for infection with ruminant nematodes, either in the condition of close contact with sheep (south-east of France) or unknown contact with ruminants (central region and French Champaign). Rabbits and hares were poorly infected with sheep digestive tract nematodes: Trichostrongylus colubriformis (second record) was encountered in one site and Trichostrongylus capricola (a first record) in another site, at very low intensity. Small sheep lungworms (Neostrongylus linearis and Cystocaulus ocreatus) were not recorded in rabbits grazing common pasture with sheep. Although transmission remained limited between lagomorphs and ruminants, it could play a role in the transfer of nematode isolates resistant to anthelmintics from one farm to another.

Inactivation of eggs and larvae of the cattle nematodes Ostertagia ostertagi and Cooperia oncophora after passage in pigs.

The study investigated the effect of gastrointestinal passage in pigs on free-living stages of bovine nematodes. Two Landrace x Yorkshire pigs, A and B, were fed fresh eggs of Ostertagia ostertagi and Cooperia oncophora while two other pigs, C and D, were fed third stage larvae (L3) of the same parasites. Faeces from the pigs were collected for 48 h after ingestion. In pigs A and B, 15 and 66% of the eggs were recovered after passage, respectively. However, only 0.003 and 0.002% of the ingested eggs developed into third stage larvae (L3) after subsequent culturing. In pigs C and D, 0.01 and 0.02% of the L3 survived the passage of the gastrointestinal tract. Fresh O. ostertagi and C. oncophora eggs were cultured in parasite free porcine and bovine faeces. Only 0.05% L3 developed in porcine faeces, whereas 21% of the eggs developed into L3 in the bovine culture. Our results demonstrate an extremely poor rate of development and survival of both bovine nematode eggs and infective larvae after passage in pigs. It may imply that pigs can play an important role in reducing transmission of cattle nematodes if the two species are grazed together or alternately.

Reduced splash dispersal of bovine parasitic nematodes from cow pats by the dung beetle Diastellopalpus quinquedens.

From a thoroughly mixed portion of cattle feces with Cooperia sp. eggs, 1-kg artificial pats were placed in 6 buckets containing 6 kg of soil each. Ten dung beetles, Diastellopalpus quinquedens, were added to each of 3 buckets. The remaining 3 buckets served as controls without beetles. When infective parasite larvae (L3) had developed in the cow pats indoors, the following procedure was followed. During occasions of rainfall each bucket was placed outdoors in the center of a wider and higher container. When the rain stopped all buckets were brought indoors, and infective larvae spread by splash droplets during the rain were collected in the containers and counted. After 33 days, the remaining dung on the soil surface in buckets with dung beetles constituted only 38% of that in the controls. Moreover, the number of L3 in feces left on the soil surface in the buckets with beetles was reduced by 88%, presumably due to beetle activity. This may explain the 70-90% reduction in splash dispersal of L3 of Cooperia sp. from cow pats attacked by beetles. The dung-burying activity of the beetles did not result in increased numbers of L3 in the soil under the cow pats, suggesting that many of the parasites in buried feces were destroyed.

The significane of Nematodirus helvetianus eggs which have survived on a pasture throughout the winter in the transmission of infection to calves.

A field experiment was carried out to determine the significance of Nematodirus helvetianus eggs which had survived throughout the winter in the transmission of infection to calves turned on to a pasture in the early summer. The calves became infected but did not exhibit symptoms of clinical disease. It was concluded that eggs which survive the winter are unlikely to be associated with outbreaks of disease.

A new host record of Camelostrongylus mentulatus (Nematoda; Trichostrongyloidea) from abomasum of a giraffe at a zoo in Japan.

Camelostrongylus mentulatus (Railliet et Henry, 1909) Orloff, 1933 (Nematoda; Trichostrongyloidea) was found from the abomasum of a three-year-old female cape giraffe, Giraffa camelopardalis giraffa, born and died in a zoo park in Yamaguchi prefecture, Japan. This is the new host record from Giraffidae and geographical distribution of C. mentulatus. Present case of C. mentulatus might be infected from other ruminants, e.g., camels, antelopes and goats, kept at a same paddock in the zoo. Risk of imported parasitic diseases by the zoo animals from outside of Japan is discussed.

Prevalence of Muellerius capillaris in free-ranging spotted deer (Cervus axis) in India and its experimental cross-transmission to goats.

A survey to assess the prevalence of parasitic infections among axis deer (Cervus axis) in three National Parks in India revealed infections with the lungworm Muellerius capillaris. Clinical signs were not evident in infected animals. Therefore, it is suggested that C. axis is probably a carrier of the infection. Under laboratory conditions, terrestrial molluscs (Macrochalamys sp.) were infected with first stage larvae of M. capillaris collected from fecal pellets of C. axis. Feeding of third stage larvae collected from these snails on day 14 post exposure produced patent infections in goats. On day 31 post infection, adult M. capillaris could be collected from the lungs of infected goats. This study establishes the possibility of cross-transmission of M. capillaris between wild and domestic animals in India.

Seasonal transmission of bovine gastrointestinal nematodes in the Texas Gulf Coast.

Sentinel calves were placed in pastures for 1 month in two environmentally dissimilar areas of the Texas Gulf Coast to determine the seasonal transmission of various gastrointestinal nematodes. Transmission was determined for Cooperia spp, Haemonchus placei, Ostertagia ostertagi, and Trichostrongylus axei. Large numbers of Cooperia spp were acquired from May through November, with the peak of transmission occurring in July and August. Haemonchus placei was encountered on both field sites but was transmitted in large numbers only at one site, during August. Ostertagia ostertagi was acquired primarily from November through May, with the peak of transmission occurring in January and February. January through March was the period when the greatest numbers of Ostertagia larvae undergoing arrested development were acquired. Trichostrongylus axei was abundance in December and January at one field site. In general, trends of transmission were the same in both areas, indicating that weather conditions were most important than vegetation type in larval transmission.

Nematodirus battus infection in calves.

In studies on the control of parasitic gastroenteritis in calves and sheep, involving an annual rotation of pastures grazed by these host species, it was shown that young cattle could play an important role in the epidemiology of Nematodirus battus, a species usually regarded as a parasite of lambs. Thus, young cattle readily acquired heavy burdens of N battus in spring and the contamination of pastures with eggs from these infections resulted in significant populations of larvae on the herbage, which were infective to both calves and lambs grazed on these pastures in the following year. Although the majority of the N battus eggs hatched in the spring, some hatched in the autumn. The calves developed a strong immunity to N battus during the grazing season as demonstrated by the absence of worms at necropsy in the autumn, despite the presence of infective larvae on the pasture.

A study on the transmission of Oesophagostomum dentatum and Hyostrongylus rubidus among outdoor reared pigs in Denmark.

This study was carried out to obtain basic information on the transmission of Oesophagostomum dentatum and Hyostrongylus rubidus in outdoor reared pigs in Denmark. Eighteen 10 weeks old worm-free pigs were allocated into 3 groups of 6 pigs each. In May, all pigs were turned out on the same parasitologically naive pasture, and after 2 weeks the pigs in groups 2 and 3 were experimentally infected with 10,800 O. dentatum and 8,700 H. rubidus infective larvae, respectively. Pigs in group 1 served as non-infected controls. All pigs were reared together on the experimental pasture for further 134 days until slaughter in October. Strongyle egg counts, differentiation of infective larvae at species level, serum pepsinogen, and herbage larval infectivity were monitored at regular intervals throughout. Both strongyle species established in the originally parasite-free pigs (group 1) and cross infections were established in group 2 and 3. The pigs were exposed to steadily increasing herbage infectivity of both species of strongyles. At the end of the experiment, geometric mean worm burdens of O. dentatum in groups 1, 2 and 3 were 1202, 6136 and 1431 respectively, the burden in group 2 being significantly higher (p < 0.05) than that of the 2 other groups. The geometric mean worm burdens of H. rubidus in groups 1, 2 and 3 were 4907, 3679 and 5246 respectively, showing no significant differences between groups.