Strongylid infection varies with age, sex, movement and social factors in wild African elephants.

Comparing parasitic infection among individuals of wildlife populations can provide insight into factors that influence wildlife disease ecology. Strongylids are parasitic worms that infect the intestinal tract of vertebrates, and infection with strongylids can be approximated by counting strongylid eggs in dung samples. Here we tested for correlations between strongylid egg counts and 18 different individual characteristics, environmental and social factors in individually known wild African elephants. We counted more eggs in the dung samples of younger elephants and females relative to mature elephants and males. We also found that elephants spending more time outside reserves shed more strongylid eggs than elephants that were more often within reserves. Elephants that were less socially integrated, as measured by how much aggression they received from other elephants, shed fewer strongylid eggs; relatedly, socially isolated orphan elephants that had left their family shed fewer strongylid eggs than elephants that remained with their family. Our results suggest that landscapes altered by livestock grazing and social disruption caused by humans may impact parasitic infection in wildlife.

Managing anthelmintic resistance in cyathostomin parasites: Investigating the benefits of refugia-based strategies.

Selective anthelmintic therapy has been recommended as a sustainable strategy for cyathostomin control in horse populations for several decades. The traditional approach has been to determine strongyle fecal egg counts (FEC) for all horses, with treatment only recommended for those exceeding a predetermined threshold. The aims are to achieve a reduction of overall egg shedding, while leaving a proportion of the herd untreated, which lowers anthelmintic treatment intensity and reduces selection pressure for development of anthelmintic resistance. This study made use of the cyathostomin model to evaluate the influence of treatment strategies with between 1 and 8 yearly treatment occasions, where either 1) all horses were treated, 2) a predetermined proportion of the herd remained untreated, or 3) horses were treated if their FEC exceeded thresholds between 100 and 600 strongyle eggs per gram. Weather data representing four different climatic zones was used and three different herd age structures were compared; 1) all yearlings, 2) all mature horses 10-20 years old, and 3) a mixed age structure of 1-20 years of age. Results indicated a consistent effect of age structure, with anthelmintic resistance developing quickest in the yearling group and slowest among the mature horses. Development of anthelmintic resistance was affected by treatment intensity and selective therapy generally delayed resistance. Importantly, the results suggest that the effects of selective therapy on resistance development are likely to vary between climatic zones and herd age structures. Overall, a substantial delaying of resistance development requires that the average number of treatments administered annually across a herd of horses needs to be about two or less. However, results also indicate that an age-structured prioritisation of treatment to younger horses should still be effective. It appears that a 'one-size-fits-all' approach to the management of anthelmintic resistance in cyathostomins is unlikely to be optimal.

Parasite prevalence increases with temperature in an avian metapopulation in northern Norway.

Climate and weather conditions may have substantial effects on the ecology of both parasites and hosts in natural populations. The strength and shape of the effects of weather on parasites and hosts are likely to change as global warming affects local climate. These changes may in turn alter fundamental elements of parasite-host dynamics. We explored the influence of temperature and precipitation on parasite prevalence in a metapopulation of avian hosts in northern Norway. We also investigated if annual change in parasite prevalence was related to winter climate, as described by the North Atlantic Oscillation (NAO). We found that parasite prevalence increased with temperature within-years and decreased slightly with increasing precipitation. We also found that a mild winter (positive winter NAO index) was associated with higher mean parasite prevalence the following year. Our results indicate that both local and large scale weather conditions may affect the proportion of hosts that become infected by parasites in natural populations. Understanding the effect of climate and weather on parasite-host relationships in natural populations is vital in order to predict the full consequence of global warming.

Presence of Kalicephalus subulatus Molin, 1861 (Nematoda, Diaphanocephalidae) in Wagler's snake, Xenodon merremi from Argentina

This is the first record of the nematode parasite Kalicephalus subulatus (Strongylida, Diaphanocephalidae) in Wagler's snakes, Xenodon merremi (Squamata, Colubridae), collected in Corrientes, Argentina. The species was observed using light and scanning electron microscope (SEM) and compared to previous studies from Tropical America. As for the metric characters, the specimens collected in Argentina are smaller than all of those found so far in other hosts. However, despite the differences in the measurements, some characters such as the dorsal lobe type III and the amphidelphic uterus allow its specific identification. Previously, this genus was found in X. merremi from Argentina but the absence of males precluded their specific identification.

Infection levels of protostrongylid nematodes in definitive caprine and intermediate gastropod hosts from Uzbekistan.

Morphological analysis of lungworms collected among Caprinae from Uzbekistan resulted in the identification of four species of Protostrongylidae: Protostrongylus rufescens, Protostrongylus hobmaieri, Spiculocaulus leuckarti and Cystocaulus ocreatus. The following species were recorded as definitive hosts: Ovis aries, Ovis ammon, Ovis vignei, Capra hircus, Capra falconeri and Capra sibirica. The prevalence of P. rufescens reached 45.3%, followed by S. leuckarti and C. ocreatus with 31.7% and P. hobmaieri with 16.9%. The sex ratio ranged between 1:3.1 and 1:6.2, with P. hobmaieri showing the strongest predominance of females over males. The prevalence of infection of small ruminants with protostrongylid nematodes increased with the age of the hosts. Protostrongyles use terrestrial gastropods as intermediate hosts, and infective larvae were found in the species Vallonia costata, Gibbulinopsis signata, Pupilla muscorum, Pseudonapaeus albiplicata, Pseudonapaeus sogdiana, Leucozonella ferghanica, Xeropicta candacharica, Candaharia levanderi and Macrochlamys sogdiana. Xeropicta candacharica was the most abundant gastropod and had the highest prevalence of infection with protostrongylids. Adult X. candacharica had a significantly higher infection intensity than juveniles. The epidemiology of protostrongylid infections is dynamic and subject to considerable changes. Further characterization of the interaction of protostrongylid parasites with their terrestrial gastropods as intermediate hosts and Caprinae as definitive hosts is required to understand these processes and to monitor the effects of changing ecological contexts.

Practical and low cost whole-organism motility assay: A step-by-step protocol.

Here, we provide a step-by-step protocol for a practical and low cost whole-organism assay for the screening of chemical compounds for activity against parasitic worms. This assay has considerable advantages over conventional methods, mainly in relation to ease of use, throughput, time and cost. It is readily suited to the screening of hundreds to thousands of compounds for subsequent hit-to-lead optimisation, and should be applicable to many different parasites and other organisms commensurate with the size of wells in the microtiter plates used for phenotypic screening.

Efficacy of selected anthelmintic drugs against cyathostomins in horses in the federal state of Brandenburg, Germany.

Cyathostomins are currently the most common internal parasites of horses. With the intensive use of anthelmintic drugs over the past decades, resistance of cyathostomins to anthelmintics is becoming a growing problem in many countries. The aim of this study was to assess the current situation on horse farms in the German federal state of Brandenburg. A pre-selected population of horses from 24 premises that had shown a prevalence of cyathostomins higher than the average in a previous study was examined for anthelmintic efficacy. Faecal egg count reduction tests (FECRTs) were performed for ivermectin (IVM) and pyrantel (PYR). For IVM, the egg reappearance period (ERP) was also examined, as a shortened ERP can be indicative of developing resistance. The efficacy of IVM on cyathostomins was high: 99.1 % of 224 horses had a zero egg count 14 days after treatment. No shortening of the ERP was detected. For the data of the FECRT for PYR, three different methods of calculation were employed: (a) the method as recommended by the World Association for the Advancement of Veterinary Parasitology (WAAVP), (b) a bootstrapping method and (c) a Markov chain Monte Carlo method. Two methods of interpretation for these data were used: Resistance was declared (a) when FECR was <90 % and the lower 95 % confidence interval (LCL) <80 % and (b) when additionally the upper 95 % confidence level (UCL) was <95 %. When applying the first interpretation, resistance against PYR was found on four yards, while, when considering the UCL, all three methods for calculation only detected resistance on one single yard. Twelve species of cyathostomins were detected in larval cultures derived from strongyle egg positive faecal samples collected 14 days after treatment with PYR by reverse line blot hybridization (RLB). In order to generate comparable data, it is suggested to establish international standards for the calculation of FECRT data.

IL-17 and neutrophils: unexpected players in the type 2 immune response.

The study of immunity to helminth infection has been central to understanding the function of type 2 cytokines and their targets. Although type 2 cytokines are considered anti-inflammatory and promote tissue repair, they also contribute to allergy and fibrosis. Here, we utilise data from helminth infection models, to illustrate that IL-17 and neutrophils, typically associated with pro-inflammatory responses, are intimately linked with type 2 immunity. Neutrophils work with IL-4Rα-activated macrophages to control incoming larvae but this comes at a cost of enhanced tissue damage. Chitinase like proteins (CLPs) bridge these diverse outcomes, inducing both protective IL-17 and reparative Th2 responses. Dysregulation of CLPs, IL-17 and neutrophils likely contribute to disease severity and pathology associated with type 2 immunity.

Developmental stage of strongyle eggs affects the outcome variations of real-time PCR analysis.

Strongyle and trichostrongyle parasites are ubiquitous nematodes of grazing livestock. Several molecular diagnostic tests are based upon measuring and quantifying DNA obtained from parasite eggs. It is well known that such eggs undergo development during storage, but it remains unknown to which extent developmental stages can affect the variation of diagnostic test results. This study investigated the influence of developmental stages of strongyle eggs on the variation real-time polymerase chain reaction (PCR) results. Mixed species strongyle eggs were obtained from the faeces of a naturally infected horse. Eggs were isolated and placed in microtiter plates with demineralized water. A total of 25 wells containing 100 eggs each were set up and kept refrigerated for up to five days. Once daily, five wells were examined on an inverted microscope at 100× magnification, where the developmental stages of the eggs were noted, and then eggs harvested for DNA extraction. The protocol was repeated three times. Genomic DNA was extracted using a commercial kit previously validated for strongyle type eggs. PCR reactions were performed with a primer set specific for the ribosomal DNA region for all strongyle type parasites (NC1, NC2). SYBR Green Real-Time PCRs were performed in triplicates. Results revealed a statistically significant increase in PCR yield after three days, which was statistically associated with beginning embryonation of the eggs. In conclusion, storage time and developmental stage of strongyle eggs are significant sources of error in studies based on quantitative real-time PCR analysis. This study suggests that for refrigerated storage of more than three days, eggs should be inactivated and preserved for further analysis.

Development of a modified molecular diagnostic procedure for the identification and quantification of naturally occurring strongylid larvae on pastures.

A molecular procedure was developed to detect and quantify larvae of different strongylid parasite species recovered from pasture samples. Two lamb flocks (L and S) grazed separate paddocks with different natural larvae challenges (one low [Paddock L] and one high [Paddock S] challenge) on a commercial farm in Western Australia. Pasture samples were collected and analysed for larvae on 9 separate occasions from each paddock. Pregnant Merino ewes were sampled on 3 separate occasions (2 pre-partum and 1 post-partum). Following lambing, 203 female crossbred lambs were identified, from which faecal samples were collected across five separate samplings. Lamb production and faecal attributes were recorded. Genomic DNA was extracted directly from lamb faeces, in addition to the genomic DNA extracts from strongylid larval species recovered from pastures. Faecal worm egg counts (FWECs) were undertaken. Species-specific qPCRs and conventional PCRs (ITS-2 nuclear ribosomal DNA) were used to screen samples for strongylid species (Teladorsagia circumcincta, Trichostrongylus spp., Haemonchus contortus, Chabertia ovina and Oesophagostomum venulosum). Negative correlations (r(2)>0.91) were found between qPCR C(q) values and log-transformed pasture larval counts for Trichostrongylus spp. and T. circumcincta. Moderate levels of agreement between pasture larval counts and qPCR results were observed (67%). A clear difference in pasture larval challenge levels was observed between the two flocks using both qPCR and conventional pasture larval counts. It is difficult to draw conclusions on the production performances of lambs from the two experimental flocks, as no further replicates were able to be conducted following this experiment. Flock L had higher dressing percentages than Flock S (P=0.038), along with significantly higher faecal consistency and breech fleece faecal soiling scores at successive samplings. The molecular procedures utilised in this study have the potential to be beneficial for livestock grazing management strategies and parasite surveillance, however further investigation is necessary before they can become part of routine diagnostics.

[Strongylids (Nematoda: Strongylidae) of domestic horses in Ukraine: modern state of Fauna and structure of the parasite community].

Species composition and structure of the strongylid community was studies on helminthological material collected from 162 domestic horses from 11 regions of Ukraine by the in vivo method. Animals were treated with anthelmintic drug "Univerm" (0.2% aversectin C, Russia). Faecal samples (200 g each) were collected from every horse at 24, 36, 48 and 60 hours after treatment; all nematodes expelled (90.851 specimens) were collected and identified. Thirty-three strongylid species from 12 genera (8 species of subfamily Strongylinae and 25--of Cyathostominae) were found in domestic horses in Ukraine. Cyathostominae dominated in the strongylid community; they were found in 100 % horses and composed 98.21% of community. "Core" of the strongylid community was composed by 7 cyathostome species. Decreasing of proportion of Strongylinae in the community for last 40 years was registered; strongylines were found in 37.6% of horses and composed 1.25% of community. Maximal prevalence was 20.98% (Strongylus vulgaris). Bray-Curtis cluster analysis revealed high similarity of strongylid communities in horses from various regions of Ukraine. Difference in general structure of strongylid communities of horses from different horse-keeping conditions was established. Horses from farms with stable-paddock keeping conditions had bimodal strongylid community structure; while horses from stable-pasture keeping conditions possessed multimodal community structure.

Synergistic effects of seasonal rainfall, parasites and demography on fluctuations in springbok body condition.

1. Seasonality of rainfall can exert a strong influence on animal condition and on host-parasite interactions. The body condition of ruminants fluctuates seasonally in response to changes in energy requirements, foraging patterns and resource availability, and seasonal variation in parasite infections may further alter ruminant body condition. 2. This study disentangles the effects of rainfall and gastrointestinal parasite infections on springbok (Antidorcas marsupialis) body condition and determines how these factors vary among demographic groups. 3. Using data from four years and three study areas, we investigated (i) the influence of rainfall variation, demographic factors and parasite interactions on parasite prevalence or infection intensity, (ii) whether parasitism or rainfall is a more important predictor of springbok body condition and (iii) how parasitism and condition vary among study areas along a rainfall gradient. 4. We found that increased parasite intensity is associated with reduced body condition only for adult females. For all other demographic groups, body condition was significantly related to prior rainfall and not to parasitism. Rainfall lagged by two months had a positive effect on body condition. 5. Adult females showed evidence of a 'periparturient rise' in parasite intensity and had higher parasite intensity and lower body condition than adult males after parturition and during early lactation. After juveniles were weaned, adult females had lower parasite intensity than adult males. Sex differences in parasitism and condition may be due to differences between adult females and males in the seasonal timing of reproductive effort and its effects on host immunity, as well as documented sex differences in vulnerability to predation. 6. Our results highlight that parasites and the environment can synergistically affect host populations, but that these interactions might be masked by their interwoven relationships, their differential impacts on demographic groups, and the different time-scales at which they operate.

Prevalence and risk factors for intestinal parasite infection in goats raised in Nakhon Pathom Province, Thailand.

The objectives of this study were to determine the prevalence and risk factors for intestinal parasite infection in goats raised in Nakhon Pathom Province. Fecal samples from 190 goats in 12 herds were collected per rectum. Questionnaires focusing on the general information about the farm and farmers, and management practices on farms were completed. Modified McMaster counting technique was performed to detect eggs of intestinal parasites. Each egg found was classified into three groups of parasites based on its characteristics. Individual and herd prevalence were calculated. Multiple logistic regressions were performed to analyze risk factors of infection. Herd prevalence of intestinal parasite infection was 100%, while individual prevalence was 79.47%. Strongyle group was the most common intestinal parasite found in this study. Moniezia spp. (8/190) and Trichuris spp. (1/190) were also found. The average number of eggs found was 1,176 eggs per gram of feces. Risk factors for intestinal parasite infection were housing system, deworming interval, and type of goat in herd. Goats housed in groups had higher infection rates compared with goats housed individually (odds ratio (OR) = 6.34; P value = 0.009). Goats in herds in which anthelmintic drugs were administered in intervals of greater than 3 months were more likely to become infected with intestinal parasites (OR = 33.07; P value <0.001). Goats in herds that kept only dairy goats were less likely to become infected than herd that kept only meat goats or kept both dairy and meat goats (OR = 46.20 and 8.75; P value <0.001 and 0.011, respectively).

Species composition and morphology of protostrongylids (Nematoda: Protostrongylidae) in ruminants from Bulgaria.

Lungs of 52 ruminants from different regions of Bulgaria, 16 from goats (Capra aegagrus f. domestica L.), 15 from sheep (Ovis ammon f. domestica L.), 11 from mouflons (Ovis musimon L.), and 10 from chamois (Rupicapra rupicapra L.), were investigated. The aim of the study was to determine the species composition of small lungworms in these hosts. The obtained results are summarized with those of previous studies, and a picture of the present status of the species composition of protostrongylids in ruminants from Bulgaria is forwarded. Morphometric data about the species Muellerius capillaris, Cystocaulus ocreatus, Neostrongylus linearis, Protostrongylus brevispiculum, and Protostrongylus rufescens are presented. The data on the morphology of these five species are supplied for the first time both for Bulgaria and the south-east part of the European continent.

Efficacy of an in-feed formulation containing ivermectin for the control of intestinal strongyles in captive zebras (Equus burchelli (Gray, 1824)).

The efficacy of medicated food pellets (containing 10mg of ivermectin per kg, UNIFEED, Veronesi, Verona, Italy) was evaluated for the control of intestinal strongyles in a group of captive zebras (Equus burchelli) at the Safari Park, Fasano (Apulia region, Brindisi Province, southern Italy). The egg reappearance period and the faecal egg counts in terms of eggs per gram of faeces were investigated. From day 0 until day +9, 30 zebras were fed with medicated pellets and pooled faecal samples were collected from the floor of paddocks. The drug showed an efficacy of 100% for up to 78 days post-treatment, with one exception. Strongyle eggs collected prior to the treatment were used to perform coprocultures and larvae harvested were molecularly identified as Cylicostephanus minutus and Cylicocyclus leptostomum with a reverse line blot hybridization assay. The administration of ivermectin with medicated food pellets was effective in controlling intestinal strongylosis in captive zebras. The opportunity to treat captive ungulates with an in-feed anthelmintic is discussed in relation to the fractious nature of these animals, which often impairs helminth control programs in zoo-parks.

Pathological findings in the caeca of naturally infected ostriches, Struthio camelus Linnaeus, 1758 (Aves, Struthionidae) parasitized by Codiostomum struthionis (Horst, 1885) Railliet and Henry, 1911 (Nematoda, Strongylidae).

In order to characterize lesions associated with Codiostomum struthionis in ostriches, 10 caeca were examined on both macro- and microscopic levels. Parasites were found in the distal third of the caecum and characterized as C. struthionis. Thickened mucosa was identified macroscopically where parasites were observed in high concentrations. Nodular areas were also observed in the distal third of the infected caeca, as well as hemorrhagic areas abutting small ulcers surrounded by edema. These findings were not observed in healthy controls. The concentration of C. struthionis found in infected animals was directly correlated with the severity of lesions observed in each caecum. These results allowed us to infer that C. struthionis is responsible for lesions in ostrich caeca.

Hatching behaviour of Nematodirus filicollis in a flock co-infected with Nematodirus battus.

The development and survival of larvae of Nematodirus filicollis, an important nematode parasite of ruminants, were extensively investigated in the laboratory for the first time, using eggs harvested from sheep grazing in the UK in winter. Third-stage larvae (L3s) developed within the eggs between 12 and 25 degrees C, with the proportion of eggs developing successfully being highest at the upper end of this range. Eggs required chilling below 10 degrees C before being able to hatch at all, and maximum hatch was observed only following chilling to 4 degrees C. Hatching occurred above 6 degrees C, and peaked at 13 degrees C, with the proportion of eggs hatching decreasing to 17-20 degrees C, which appears to form an upper threshold temperature for hatching. When fully developed eggs were placed at 11-13 degrees C, L3 emerged very rapidly with hatching being completed within 6 days. Mortality of emerged L3s increased with temperature, particularly above 20 degrees C. For this parasite, given the low minimum hatching thresholds and the rapid emergence of L3, the existence of an upper hatching threshold is likely to have limited impact on patterns of larval availability on pasture. The strict requirement for chilling is surprising, being at odds with both previous observations and the wide geographical distribution of this species. It is possible that the proportion of eggs requiring chilling is an adaptable trait, enabling early infection of naïve hosts in the face of competition from N. battus, yet also persistence in regions that do not provide good conditions for the chilling of eggs. Further study of selectable variation in this trait could help us to understand the contrasting geographical ranges and disease patterns within the genus, and how different species might respond to climate change.

[On the occurrence of the nematode Oswaldocruzia filiformis (Strongylida: Molineidae) in Karelia].

Contents of the intestines of the viper Vipera berus (L., 1758) from Kizhi archipelago (Lake Onego) was examined. Helminth fauna of the viper was found to include single nematode species, Oswaldocruzia filiformis Goeze, 1782 (Strongylida: Molineidae). Prevalence of the invasion was 60%, intensity of the invasion was 1-8 specimens per host, index of abundance was 1.92 specimens. Measurements and pictures of the parasite are given. Morphometric data on the nematode from viper are compared with those from other host species.

Phenotypic analysis of host-parasite interactions in lambs infected with Teladorsagia circumcincta.

Female Blackface lambs expected to exhibit genetic variability for resistance to gastrointestinal nematodes, were either exposed to continuous experimental infections of Teladorsagia circumcincta or were sham-dosed to monitor phenotypic responses to infection. As a measure of parasitism and host response, worm-eggs in faeces (faecal egg count, FEC) were counted over a 3-month period and worm burdens were ascertained at post-mortem. The host response to the infection was also measured by differential counts of white blood cells, anti-T. circumcincta IgA antibody levels and body weight. Results suggest that nematode abundance (mean number of parasites per host) and prevalence (proportion of infected animals) were maximal shortly after the beginning of infection (21 days p.i.) when virtually all the infected animals were shedding worm eggs. Increasing anti-T. circumcincta IgA antibody and eosinophil concentrations were associated with a reduction in total numbers of adult worms and an increase in the frequency of early L4s. The data also suggest that genetic selection for an enhanced anti-T. circumcincta IgA response might complement selection based on a reduced FEC as a strategy to select for resistance to gastrointestinal nematodes.