Two new species of the genus Sectonema Thorne, 1930 (Nematoda, Dorylaimida, Aporcelaimidae) from Iran, with new insights into its evolutionary relationships.

Two new species of the genus Sectonema found in northern Iran are characterized, including morphological descriptions and molecular (18S-, 28S-rDNA) analyses. Sectonema tehranense sp. nov. is distinguished by its 7.22 - 8.53 mm long body, lip region offset by constriction and 24 - 31 µm wide with perioral lobes and abundant setae- or cilia-like projections covering the oral field, mural tooth 15.5 - 17 µm long at its ventral side, neck 1091 - 1478 µm long, pharyngeal expansion occupying 61 - 71% of the total neck length, female genital system diovarian, uterus simple and 3.9 - 4.2 times the corresponding body diameter long, transverse vulva (V = 49 - 59), tail short and rounded (44 - 65 µm, c = 99 - 162, c' = 0.6 - 0.8), spicules 111 - 127 µm long, and 7 - 10 spaced ventromedian supplements with hiatus. Sectonema noshahrense sp. nov. displays a 4.07 - 4.73 mm long body, lip region offset by constriction and 23 - 25 µm wide with perioral lobes and abundant setae- or cilia-like projections covering the oral field, odontostyle 14 - 14.5 µm long, neck 722 - 822 µm long, pharyngeal expansion occupying 66 - 68% of the total neck length, female genital system diovarian, uterus simple and 2.4 - 2.7 times the corresponding body diameter long, transverse vulva (V = 54 - 55), tail convex conoid (39 - 47 µm, c = 91 - 111, c' = 0.8 - 0.9), spicules 82 µm long, and seven spaced ventromedian supplements with hiatus. Molecular analyses confirm a maximally supported (Epacrolaimus + Metaporcelaimus + Sectonema) clade and a tentative biogeographical pattern, with sequences of Indolamayan taxa forming a clade separated from those of Palearctic ones. Parallel or convergent evolution processes might be involved in the phylogeny of the species currently classified under Sectonema. This genus is certainly more heterogeneous than previously assumed.

A new species of the genus Hexamermis Steiner, 1924 (Nematoda: Mermithidae) from northern Iran: a nematode with an unusual uterine morphology.

Hexamermis zirabi sp. n., recovered from a natural habitat of Mazandaran province, north of Iran, is described based on morphological and molecular data. The new species is characterized by its six cephalic papillae; cuticle with distinct cross fibers; conoid or sharply tapered head; mouth terminal; six hypodermal cords; J-shaped vagina oriented to the anterior end of body; uterus with Z-organs or sclerotized bodies; tail similar in both sexes and bluntly rounded; spicules paired, separate, slightly curved, shorter than body width at cloaca, with rounded tip; and male genital papillae arranged in five rows. In addition to the morphological study, molecular phylogenetic analyses using a partial large subunit (28S D2-D3) were also performed, and the new species formed a highly supported (1.00% Bayesian posterior probability (BPP)) clade with Hexamermis popilliae.

Heterocheilus floridensis sp. n. (Nematoda: Heterocheilidae) from the West Indian manatee Trichechus manatus (Trichechidae, Sirenia) in Florida, USA.

Morphological data are used to describe a new nematode species, Heterocheilus floridensis sp. n. (Heterocheilidae) from the digestive tract of the Florida manatee Trichechus manatus latirostris (Harlan) (Trichechidae, Sirenia) from Florida, USA. Examination by light and scanning electron microscopy revealed that the new species differs from the related Heterocheilus tunicatus Diesing, 1839 mainly by having dentigerous ridges on the inner surface of the lips, a median unpaired papilla located anterior to the cloaca, and a considerably larger body size. Sequence data for subunits I and II of mitochondrial cytochrome oxidase gene, 18S small subunit and 28S ribosomal RNA genes were provided for molecular characterisation of the new species. However, the current unavailability of homologous sequence data for congeneric specimens precluded a molecular assessment of the morphological species hypothesis, and ascaridoid phylogenetic hypotheses could not be advanced. Specimens of Heterocheilus sp. collected from the Antillean manatee Trichechus manatus manatus Linnaeus in Puerto Rico, on loan from the US National Museum of Natural History, were morphologically consistent with the new species, so apparently all congeneric nematodes reported from both subspecies of the West Indian manatee Trichechus manatus Linnaeus and previously identified as H. tunicatus belong rather to H. floridensis sp. n. Heterocheilus hagenbecki (Khalil et Vogelsang, 1932) Sprent 1980 is here considered to be a species inquirenda. A key to valid species of Heterocheilus Diesing, 1839 is provided.

Conserved switch genes that arose via whole-genome duplication regulate a cannibalistic nematode morph.

Experimental genetics in a nematode reveals a key role for developmental plasticity in the evolution of nutritional diversity.

Intestinal cell diversity and treatment responses in a parasitic nematode at single cell resolution.

BACKGROUND: Parasitic nematodes, significant pathogens for humans, animals, and plants, depend on diverse organ systems for intra-host survival. Understanding the cellular diversity and molecular variations underlying these functions holds promise for developing novel therapeutics, with specific emphasis on the neuromuscular system's functional diversity. The nematode intestine, crucial for anthelmintic therapies, exhibits diverse cellular phenotypes, and unraveling this diversity at the single-cell level is essential for advancing knowledge in anthelmintic research across various organ systems. RESULTS: Here, using novel single-cell transcriptomics datasets, we delineate cellular diversity within the intestine of adult female Ascaris suum, a parasitic nematode species that infects animals and people. Gene transcripts expressed in individual nuclei of untreated intestinal cells resolved three phenotypic clusters, while lower stringency resolved additional subclusters and more potential diversity. Clusters 1 and 3 phenotypes displayed variable congruence with scRNA phenotypes of C. elegans intestinal cells, whereas the A. suum cluster 2 phenotype was markedly unique. Distinct functional pathway enrichment characterized each A. suum intestinal cell cluster. Cluster 2 was distinctly enriched for Clade III-associated genes, suggesting it evolved within clade III nematodes. Clusters also demonstrated differential transcriptional responsiveness to nematode intestinal toxic treatments, with Cluster 2 displaying the least responses to short-term intra-pseudocoelomic nematode intestinal toxin treatments. CONCLUSIONS: This investigation presents advances in knowledge related to biological differences among major cell populations of adult A. suum intestinal cells. For the first time, diverse nematode intestinal cell populations were characterized, and associated biological markers of these cells were identified to support tracking of constituent cells under experimental conditions. These advances will promote better understanding of this and other parasitic nematodes of global importance, and will help to guide future anthelmintic treatments.

In silico assessment of 18S rDNA metabarcoding markers for the characterization of nematode communities.

Nematodes are keystone actors of soil, freshwater and marine ecosystems, but the complexity of morphological identification has limited broad-scale monitoring of nematode biodiversity. DNA metabarcoding is increasingly used to assess nematode diversity but requires universal primers with high taxonomic coverage and high taxonomic resolution. Several primers have been proposed for the metabarcoding of nematode diversity, many of which target the 18S rRNA gene. In silico analyses have a great potential to assess key parameters of primers, including taxonomic coverage, resolution and specificity. Based on a recently-available reference database, we tested in silico the performance of fourteen commonly used and one newly optimized primer for nematode metabarcoding. Most primers showed very good coverage, amplifying most of the sequences in the reference database, while four markers showed limited coverage. All primers showed good taxonomic resolution. Resolution was particularly good if the aim was the identification of higher-level taxa, such as genera or families. Overall, species-level resolution was higher for primers amplifying long fragments. None of the primers was highly specific for nematodes as, despite some variation, they all amplified a large number of other eukaryotes. Differences in performance across primers highlight the complexity of the choice of markers appropriate for the metabarcoding of nematodes, which depends on a trade-off between taxonomic resolution and the length of amplified fragments. Our in silico analyses provide new insights for the identification of the most appropriate primers, depending on the study goals and the origin of DNA samples. This represents an essential step to design and optimize metabarcoding studies assessing nematode diversity.

Machine learning prediction of multiple anthelmintic resistance and gastrointestinal nematode control in sheep flocks.

The high prevalence of Haemonchus contortus and its anthelmintic resistance have affected sheep production worldwide. Machine learning approaches are able to investigate the complex relationships among the factors involved in resistance. Classification trees were built to predict multidrug resistance from 36 management practices in 27 sheep flocks. Resistance to five anthelmintics was assessed using a fecal egg count reduction test (FECRT), and 20 flocks with FECRT < 80% for four or five anthelmintics were considered resistant. The data were randomly split into training (75%) and test (25%) sets, resampled 1,000 times, and the classification trees were generated for the training data. Of the 1,000 trees, 24 (2.4%) showed 100% accuracy, sensitivity, and specificity in predicting a flock as resistant or susceptible for the test data. Forage species was a split common to all 24 trees, and the most frequent trees (12/24) were split by forage species, grazing pasture area, and fecal examination. The farming system, Suffolk sheep breed, and anthelmintic choice criteria were practices highlighted in the other trees. These management practices can be used to predict the anthelmintic resistance status and guide measures for gastrointestinal nematode control in sheep flocks.

Body size mediates the functional potential of soil organisms by diversity and community assembly across soil aggregates.

Body size is an important life-history trait that affects organism niche occupancy and ecological interactions. However, it is still unclear to what extent the assembly process of organisms with different body sizes affects soil biogeochemical cycling processes at the aggregate level. Here, we examined the diversity and community assembly of soil microorganisms (bacteria, fungi, and protists) and microfauna (nematodes) with varying body sizes. The microbial functional potential associated with carbon, nitrogen, phosphorus, and sulfur metabolism within three soil aggregate sizes (large macroaggregates, > 2 mm; small macroaggregates, 0.25-2 mm; and microaggregates, < 0.25 mm) were determined by metagenomics. We found that the smallest microbes (bacteria) had higher α-diversity and lower ß-diversity and were mostly structured by stochastic processes, while all larger organisms (fungi, protists, and nematodes) had lower α-diversity and were relatively more influenced by deterministic processes. Structural equation modeling indicated that the microbial functional potential associated with carbon, nitrogen, phosphorus, and sulfur metabolism was mainly influenced by the bacterial and protist diversity in microaggregates. In contrast, the microbial functional potential was primarily mediated by the assembly processes of four organism groups, especially the nematode community in macroaggregates. This study reveals the important roles of soil organisms with different body sizes in the functional potential related to nutrient cycling, and provides new insights into the ecological processes structuring the diversity and community assembly of organisms of different body sizes at the soil aggregate level, with implications for soil nutrient cycling dynamics.

Helminths of captive and free-ranging populations of the mountain gazelle (Gazella gazella): Evidence from faecal examination.

BACKGROUND: Understanding parasite diversity in wild and captive animal populations has critical implications for both individual animal health and ecosystem dynamics in a broader sense. In mountain gazelles (Gazella gazella), the gastrointestinal helminth community is poorly understood, limiting our efforts in the conservation of this endangered bovid species. This species has only two remaining populations in the world, including the isolated northernmost population in Türkiye. OBJECTIVES: To identify and compare the diversity and prevalence of gastrointestinal helminths in captive and free-ranging populations of mountain gazelles in Hatay, Türkiye, and to assess potential zoonotic risks. METHODS: In total, 105 fresh faecal samples, 45 individual samples and 60 faecal samples, representing 16 pools, from both captive and free-ranging populations were collected and analysed using Fulleborn flotation, Benedek sedimentation and Bearman-Wetzel methods faecal flotation methods, including the McMaster technique to determine the severity of infection. RESULTS: We detected 12 helminth taxa in our examination of faecal samples, including gastrointestinal nematodes, lungworms and trematodes. Parasites from the Trichostrongyloidea family demonstrated variable hatching stages and rates, potentially influenced by ambient conditions. We also detected one protozoan among the samples. Our results revealed a higher diversity of parasites in free-ranging populations compared to captive ones. CONCLUSIONS: This study underscores the necessity for regular parasitological surveillance in both captive and free-ranging wildlife populations for effective conservation management. It also contributes to the 'One Health' perspective by highlighting the potential zoonotic risks posed by parasites in wild ruminants. Our results have implications for the conservation and management of the mountain gazelle.

Assessment of Common Factors Associated with Droplet Digital PCR (ddPCR) Quantification of Paratrichodorus allius in Soil.

This research investigated the factors associated with the quantitative detection of Paratrichodorus allius in soil using droplet digital PCR (ddPCR). Small-sized nematodes exhibited significantly lower DNA quantities compared to their medium and large counterparts. Soil pre-treatments (room temperature drying and 37 °C oven-drying) demonstrated no substantial impact on ddPCR detection, and soil storage (0-3 months at 4 °C) exhibited negligible alterations in DNA quantities. A commercial DNA purification kit improved the resulting quality of ddPCR, albeit at the cost of a notable reduction in DNA quantity. Upon assessing the impact of inhibitors from soil extracts, a higher inhibitor concentration (5%) influenced ddPCR amplification efficiency. Incorporating bovine serum albumin (BSA) (0.2 µg/µL or 0.4 µg/µL) into the ddPCR setup mitigated the issue. In brief, while ddPCR exhibits minimal sensitivity to soil pre-treatments and storage, higher concentrations of PCR inhibitors and the DNA purification process can influence the results. Despite ddPCR's capability to detect nematodes of all sizes, quantification may not precisely reflect soil population. Incorporating BSA into the ddPCR setup enhances both detection and quantification capacities. This study represents the first comprehensive investigation of its kind for plant-parasitic nematodes, providing crucial insights for application of ddPCR in nematode diagnosis directly from the soil DNA.

Identification and Targeted Quantification of Endogenous Neuropeptides in the Nematode Caenorhabditis elegans Using Mass Spectrometry.

The nematode Caenorhabditis elegans lends itself as an excellent model organism for peptidomics studies. Its ease of cultivation and quick generation time make it suitable for high-throughput studies. The nervous system, with its 302 neurons, is probably the best-known and studied endocrine tissue. Moreover, its neuropeptidergic signaling pathways display numerous similarities with those observed in other metazoans. Here, we describe two label-free approaches for neuropeptidomics in C. elegans: one for discovery purposes, and another for targeted quantification and comparisons of neuropeptide levels between different samples. Starting from a detailed peptide extraction procedure, we here outline the liquid chromatography tandem mass spectrometry (LC-MS/MS) setup and describe subsequent data analysis approaches.

To live free or being a parasite: The optimal foraging behavior may favor the evolution of entomopathogenic nematodes.

Facultative parasites can alternate between a free-living and a parasitic existence to complete their life cycle. Yet, it remains uncertain which lifestyle they prefer. The optimal foraging theory suggests that food preferences align with fitness benefits. To test this hypothesis, we investigated the facultative parasite nematode Rhabditis regina, assessing its host preference and the associated benefits. Two experiments were conducted using wild nematode populations collected from Phyllophaga polyphylla, their natural host. In the first experiment, we used a behavioral arena to assess host preference between the natural host and two experimental hosts: Spodoptera frugiperda which is an alternative host and dead Tenebrio molitor, which simulates a saprophytic environment. In the second experiment, we subjected wild nematodes to "experimental evolution" lasting 50 generations in S. frugiperda and 53 generations in T. molitor carcass. We then compared life history traits (the size, survival, number of larvae, and glycogen and triglycerides as energy reserves) of dauer larvae with those nematodes from P. polyphylla (control group). We found a significant preference for P. polyphylla, which correlated with higher values in the nematode's life history traits. In contrast, the preference for S. frugiperda and the saprophytic environment was lower, resulting in less efficient life history traits. These findings align with the optimal foraging theory, as the nematode's parasitic preferences are in line with maximizing fitness. This also indicates that R. regina exhibits specificity to P. polyphylla and is better adapted to a parasitic lifestyle than a free-living one, suggesting an evolutionary pathway towards parasitism.

Four new records of Sipuncula (Annelida) from Northern Brazil.

There are very few taxonomic studies of Brazilian sipunculan worms, and the species occurring along the northern coasts are largely understudied. We report herein the occurrence of four shallow water species along the intertidal zones of Bahia and Pernambuco states. Antillesoma antillarum (Grbe & Oersted, 1858) occurs from underneath beach rocks or within sandstone reefs and may reach high densities. The large species Sipunculus (Sipunculus) nudus Linnaeus, 1766, Sipunculus polymyotus Fisher, 1947, and Xenosiphon branchiatus (Fischer, 1895) were collected in sandy-mud, low-energy intertidal environments. For the first time, we report that sipunculans are collected and used as fishing bait by local fishermen in the Western Atlantic. All species are fully described, including notes on their external and internal anatomy.

Deciphering the impact of endoparasitic infection on immune response and gut microbial composition of Channa punctata.

Intestinal parasitic infections caused by helminths are globally distributed and are a major cause of morbidity worldwide. Parasites may modulate the virulence, gut microbiota diversity and host responses during infection. Despite numerous works, little is known about the complex interaction between parasites and the gut microbiota. In the present study, the complex interplay between parasites and the gut microbiota was investigated. A total of 12 bacterial strains across four major families, including Enterobacteriaceae, Morganellaceae, Flavobacteriaceae, and Pseudomonadaceae, were isolated from Channa punctata, infected with the nematode species Aporcella sp., Axonchium sp., Tylencholaimus mirabilis, and Dioctophyme renale. The findings revealed that nematode infection shaped the fish gut bacterial microbiota and significantly affected their virulence levels. Nematode-infected fish bacterial isolates are more likely to be pathogenic, with elevated hemolytic activity and biofilm formation, causing high fish mortality. In contrast, isolates recovered further from non-parasitised C. punctata were observed to be non-pathogenic and had negligible hemolytic activity and biofilm formation. Antibiogram analysis of the bacterial isolates revealed a disproportionately high percentage of bacteria that were either marginally or multidrug resistant, suggesting that parasitic infection-induced stress modulates the gut microenvironment and enables colonization by antibiotic-resistant strains. This isolation-based study provides an avenue to unravel the influence of parasitic infection on gut bacterial characteristics, which is valuable for understanding the infection mechanism and designing further studies aimed at optimizing treatment strategies. In addition, the cultured isolates can supplement future gut microbiome studies by providing wet lab specimens to compare (meta)genomic information discovered within the gut microenvironment of fish.

Nematodes associated with terrestrial gastropod molluscs in Belgium and additional characterisation of Pellioditis californica and P. hermaphrodita.

A survey for slug- and snail-associated nematodes was conducted in forests, parks, botanical gardens, and nature reserves at 13 localities in Belgium to uncover more diversity of gastropod mollusc-associated nematodes and to characterise Pellioditis populations found in the country. A total of 319 slugs and snails belonging to nine species were examined. Arion vulgaris was the most commonly found mollusc species in this study (eight locations), and 19.4% of the examined mollusc specimens were found infected by nematodes. The highest prevalence of nematodes was observed in Cornu aspersum (60%) followed by A. vulgaris (34.8%), Limax maximus (28.6%), and Cepaea sp. (20%). Eleven nematode species belonging to eight families were isolated and identified from the mollusc hosts including Alloionema appendiculatum, Angiostoma dentiferum, A. gandavense, Angiostrongylus vasorum, Cosmocerca longicauda, Panagrolaimus cf. subelongatus, Pellioditis californica, P. hermaphrodita, Rhabditis sp., Tetrameres cf. fissispina, and Troglostrongylus cf. brevior.Pellioditis was the most commonly found nematode genus (at nine localities) and C. longicauda and P. californica were reported in Belgium for the first time. Co-infections of more than one nematode species were observed in eight (2.5%) molluscs specimens. Most co-infections consisted of two nematode species. In one A. vulgaris specimen, a co-infection of three nematode species (A. vasorum, P. hermaphrodita, and Tetrameres cf. fissispina) was observed. Four ex vivo cultures of P. californica and six ex vivo cultures of P. hermaphrodita were established from single hermaphrodites, and both species were described based on light microscopy, scanning electron microscopy, and morphometric, morphological, and molecular data.

Acquisition and transmission of Grapevine fanleaf virus (GFLV) by Xiphinema index and Xiphinema italiae (Longidoridae).

Grapevine fanleaf virus (GFLV) is one of the most severe virus diseases of grapevines, causing fanleaf degeneration that is transmitted by Xiphinema index. This paper aims to isolate Xiphinema species from Tunisian vineyard soil samples and assess their ability to acquire and transmit GFLV under natural and controlled conditions. Based on morphological and morphometric analyses, Tunisian dagger nematodes were identified as X. index and Xiphinema italiae. These results were confirmed with molecular identification tools using species-specific polymerase chain reaction primers. The total RNA of GFLV was extracted from specimens of Xiphinema and amplified based on real-time polymerase chain reaction using virus-specific primers. Our results showed that X. index could acquire and transmit the viral particles of GFLV. This nepovirus was not detected in X. italiae, under natural conditions; however, under controlled conditions, this nematode was able to successfully acquire and transmit the viral particles of GFLV.

A molecular assessment of Ostertagia leptospicularis and Spiculopteragia asymmetrica among wild fallow deer in Northern Ireland and implications for false detection of livestock-associated species.

BACKGROUND: Wild deer populations utilizing livestock grazing areas risk cross-species transmission of gastrointestinal nematode parasites (GINs), including GINs with anthelmintic resistance (AR) traits. Wild deer have been shown to carry problematic GIN species such as Haemonchus contortus and Trichostrongylus species in the UK, but the presence of livestock GINs in Northern Ireland deer populations is unknown. Also, is it not known whether AR traits exist among GINs of deer such as Ostertagia leptospicularis and Spiculopteragia asymmetrica in pastureland where anthelmintics are heavily used. METHODS: Adult-stage GIN samples were retrieved from Northern Irish wild fallow deer abomasa. Individual specimens were subject to a species-specific PCR analysis for common sheep and cattle GIN species with ITS-2 sequence analysis to validate species identities. In addition, the beta-tubulin gene was subject to sequencing to identify benzimidazole (BZ) resistance markers. RESULTS: ITS-2 sequencing revealed O. leptospicularis and S. asymmetrica, but species-specific PCR yielded false-positive hits for H. contortus, Teladorsagia circimcincta, Trichostrongylus axei, T. colubriformis, T. vitrinus and Ostertagia ostertagi. For beta-tubulin, O. leptospicularis and S. asymmetrica yielded species-specific sequences at the E198 codon, but no resistance markers were identified in either species at positions 167, 198 or 200 of the coding region. DISCUSSION: From this report, no GIN species of significance in livestock were identified among Northern Ireland fallow deer. However, false-positive PCR hits for sheep and cattle-associated GINs is concerning as the presence of deer species in livestock areas could impact both deer and livestock diagnostics and lead to overestimation of both GIN burden in deer and the role as of deer as drivers of these pathogens. ITS-2 sequences from both O. leptospicularis and S. asymmetrica show minor sequence variations to geographically distinct isolates. AR has been noted among GINs of deer but molecular analyses are lacking for GINs of wildlife. In producing the first beta-tubulin sequences for both O. leptospicularis and S. asymmetrica, we report no BZ resistance in this cohort. CONCLUSIONS: This work contributes to genetic resources for wildlife species and considers the implications of such species when performing livestock GIN diagnostics.

Utilisation of indigenous knowledge to control gastrointestinal nematodes in Southern Africa.

Gastrointestinal nematodes (GIN) exacerbate the impact of droughts on the survival of cattle. The inadequacies of the conventional system make it increasingly important to explore indigenous knowledge (IK) to create drought-tolerant and GIN resilient herds. The objective of the study was to assess the indigenous strategies for controlling GIN during droughts. Face-to-face interviews with experts on IK were conducted to give insight into the importance, methods and ranking of GIN control. The experts identified 86 cattle that were used to test their assertions. The control methods used were identifying cattle that were susceptible to high GIN loads using predisposing factors, diagnosis of GIN burdens using faecal appearance, and treatment using phytotherapy. Experts ranked predisposing factors as the most critical control strategy and identified body condition, class, sex, coat colour, pregnancy status and lactation status as predisposing factors to high GIN burdens. Thin, older, dark-coloured cattle, as well as pregnant and lactating cows, were considered susceptible to GIN. However, pregnancy status, coat colour and sex were significantly associated with high GIN burdens. Cows were 2.6 times more likely to have high GIN burdens than bulls. Dark-coloured cattle were 3.5 times more likely to have high GIN burdens than light-coloured ones, and the likelihood of pregnant cows was 4.9 times higher than non-pregnant cows. A dark-coloured pregnant cow was extremely susceptible to high GIN burdens. In conclusion, knowledge of predisposing factors informs selection decisions when purchasing foundation stock. Cattle that are susceptible to high GIN loads are prioritised during droughts or culled where resources are scarce.

The biocontrol nematode Phasmarhabditis hermaphrodita infects and increases mortality of Monadenia fidelis, a non-target terrestrial gastropod species endemic to the Pacific Northwest of North America, in laboratory conditions.

Inundative biological control (biocontrol) efforts in pest management lead to the mass distribution of commercialized biocontrol agents. Many 'biocontrol gone awry' incidents have resulted in disastrous biodiversity impacts, leading to increased scrutiny of biocontrol efforts. The nematode Phasmarhabditis hermaphrodita is sold as a biocontrol agent on three continents and targets pest gastropods such as Deroceras reticulatum, the Grey Field Slug; P. hermaphrodita is not presently approved for use in the United States. Investigations into the potential for P. hermaphrodita to infect non-target gastropod species of conservation relevance, however, are limited. We examined the effects of three strains of P. hermaphrodita on mortality in Monadenia fidelis, the Pacific Sideband, a snail species endemic to the Pacific Northwest of North America, in laboratory conditions. Across a 71-day laboratory infectivity assay, snails exposed to each of the three nematode strains, each analyzed at two doses, experienced a mean 50% mortality by days 20-42. All nematode-treated snails were dead by the end of the study. By contrast, 30/30 water-control snails experienced no mortality. Nematodes killed smaller, juvenile-stage snails significantly faster than those in larger and more developmentally advanced hosts. Our results provide direct evidence that the biocontrol nematode P. hermaphrodita infects and kills M. fidelis, a non-target gastropod species endemic to the Pacific Northwest, in laboratory conditions. This study suggests that introduction of P. hermaphrodita to new ecosystems might negatively impact endemic gastropod biodiversity and advocates for further investigation of non-target effects, including in conditions closer to the natural environments of non-target species.

Epidemiology and field efficacy of anthelmintic drugs associated with gastrointestinal nematodes of sheep in Nejo district, Oromia, Ethiopia.

Gastrointestinal nematodes (GINs) are major constraints to health and productivity of small ruminants. Methods of their control relies mainly on anthelmintic drugs; however, the indiscriminate use of these drugs could lead to the development of anthelmintic resistance (AR). This study aimed to investigate the epidemiology of GINs infection, and field evaluation of anthelmintic efficacy in sheep. The epidemiological data were collected using a cross-sectional study design while a farm-based field study design was employed for the evaluation of anthelminthic efficacy. Furthermore, standard parasitological techniques were employed for qualitative and quantitative worm identification. The overall prevalence indicated 50.3%. Six genera of GINs (Haemonchus, Trichostrongylus, Oesophagostomum/Chabertia, Trichuris, Teladosargia/Ostertagia and Nematodirus) were identified. Among the identified genera, Haemonchus (25.4%) and Trichostrongylus (24.8%) were the dominant genera followed by mixed infection (21.8%), Oesophagostomum/Chabertia (10.4%), Trichuris (7.8%), Teladosargia (Ostertagia) (5.7%) and Nematodirus (4.1%). Mixed infections consisted either of double infections with Haemonchus and Trichostrongylus, or triple infections with Haemonchus, Trichostrongylus and Trichuris. The McMaster egg counting results showed that the mean EPG of infected sheep was 845.6. The results also showed 66 (34.2%), 101 (52.3%) and 26 (13.5%) sheep had low, moderate and heavy worm burden, respectively. Albendazole and Ivermectin showed low efficacy (percentage reductions = 90% and 92%; 95% lower confidence limit = 82.1% and 83.6% respectively) whereas Tetramisole was effective (FECR% = 96.8%; 95% LCL = 93.4%). Factors such as age, body condition, management system and past deworming history of sheep were found to have a statistically significant (p < 0.05) influence on the occurrence and burden of the worms. This is further explained as the highest prevalence and worm burden was detected in sheep of young age (p = 0.008; OR = 0.58; 95% CI = 0.39-0.87), poor body condition (p = 0.001; OR = 0.08; 95% CI = 0.04-0.16) and sheep kept under semi-intensive (p = 0.04; OR = 1.53; 95% CI = 1.02-2.29) with no deworming history for the last two months (p = 0.001; OR = 2.97; 95% CI = 1.94-4.56). The study results revealed that nematode infections were among sheep health constraints that could hurt their productivity while low efficacy of Albendazole and Ivermectin were detected. Therefore, the appropriate management techniques of GIN infections should be designed and implemented. Moreover, a further study involving more sensitive techniques (e.g. Mini-FLOTAC, molecular, and serological techniques) should be conducted by considering different host and environmental risk factors such as production level and seasons.