Efficacious therapeutic strategies against lymphatic filariasis are always sought after. However, natural products are a promising resource for developing effective antifilarial agents. Azadirachtin, a significant tetranortriterpenoid phytocompound found in Azadirachta indica, was evaluated in vitro for antifilarial potential against the filarial parasite Setaria cervi. Dye exclusion and MTT assay confirmed the antifilarial potential of azadirachtin against S. cervi with a median lethal dose (LC50) of 6.28 μg/ml for microfilariae (mf), and 9.55 μg/ml for adult parasites. Morphological aberrations were prominent in the histological sections of the azadirachtin-exposed parasites. Moreover, alterations in the reactive oxygen species (ROS) parameters in treated parasites were evident. Induction of apoptosis in treated parasites was confirmed by DNA laddering, acridine orange (AO)/ethidium bromide (EtBr) double staining and in situ DNA fragmentation. The downregulation of anti-apoptotic CED-9 and upregulation of proapoptotic EGL-1, CED-4 and CED-3 at both the transcription and translation levels confirmed apoptosis execution at the molecular level. Changes in the gene expressions of nuc-1, cps-6 and crn-1 further clarified the molecular cause of DNA degradation. Furthermore, azadirachtin was found to be non-toxic in both in vitro and in vivo toxicity analyses. Therefore, the experimental evidence detailed the pharmacological effectiveness of azadirachtin as a possible therapeutic agent against filariasis.

This study assessed the efficacy of closantel vis-à-vis herbal extracts with known anti-parasitic properties, against fenbendazole-resistant nematodes in goats maintained under a semi-intensive system of management at the University goat farm, Jabalpur. Fifty goats were randomly assigned to five groups, each comprising 10 animals, irrespective of their breed, age and sex. Each animal in Group I, II and III was orally administered with aqueous leaf extracts of neem (Azadirachta indica) at 1 g/kg body weight, sitaphal (Annona squamosa) at 1.5 g/kg body weight and tobacco (Nicotiana tabacum) at 1 g/kg body weight, respectively, whereas Group IV was an untreated control group. Each animal in Group V was orally treated with closantel at 10 mg/kg body weight. During the course of the study, all animals were maintained under an identical semi-intensive system of management. Compared to the untreated control group (Group IV), there was no conspicuous reduction in post-treatment (day 10) faecal egg counts (FEC) in animals administered with the herbal extracts (Groups I, II and III), which is suggestive of poor anti-parasitic activity. However, using the faecal egg count reduction test (FECRT), the overall efficacy of closantel was recorded as 95.64%. This supports the rotational use of closantel as a preferred choice over the benzimidazole group of anthelmintics and/or herbal extracts to meet the acute challenge of in situ development of drug-resistant gastrointestinal nematodes, especially Haemonchus contortus.

In this study, we evaluated the in vivo comparative efficacy of ivermectin and Nigella sativa extract against helminths in Aseel chickens, and the effects of helminths on blood parameters before and after treatment in Aseel chickens. Forty naturally infected adult Aseel chickens were randomly divided into four groups (n = 10 each): group A (ivermectin at 300 μg/kg); group B (N. sativa extract at 200 mg/kg); group C (ivermectin at 300 μg/kg + N. sativa extract at 200 mg/kg); group D was kept as a positive control to monitor time-related changes. On day 28 post treatment, the mean percentages of faecal egg-count reduction (FECR %) in groups A, B and C were recorded as 93.58, 88.09 and 100.00%, respectively. Further data analysis showed significantly higher efficacy in group C (100 ± 0.00%) than in groups A and B (P < 0.001). Highly significant (P < 0.001) improvements in mean percentage values of packed cell volume (PCV %) were recorded in groups A and C on days 14 and 28 post treatment. Meanwhile, the improvements in mean values of haemoglobin (Hb) concentration in groups A, B and C were highly significant (P < 0.001) when compared to that of group D on day 28 post treatment. The synergistic combination of ivermectin and N. sativa extract possessed greater efficacy than either ivermectin or N. sativa extract used alone. Furthermore, both PCV % and Hb concentration values gradually increased in the treated groups compared to the control group, in which PCV % and Hb concentration gradually decreased throughout the trial.

Androgens have been shown to exert a cysticidal effect upon Taenia crassiceps, an experimental model of cysticercosis. To further inquire into this matter, the Taenia crassiceps model was used to evaluate the expression of several proteins after testosterone (T4) and dihydrotestosterone (DHT) in vitro treatment. Under 2-D proteomic maps, parasite extracts were resolved into approximately 130 proteins distributed in a molecular weight range of 10–250 kDa and isoelectrical point range of 3–10. The resultant proteomic pattern was analysed, and significant changes were observed in response to T4 and DHT. Based on our experience with electrophoretic patterns and proteomic maps of cytoskeletal proteins, alteration in the expression of isoforms of actin, tubulin and paramyosin and of other proteins was assessed. Considering that androgens may exert their biological activity in taeniids through the non-specific progesterone receptor membrane component (PGRMC), we harnessed bioinformatics to propose the identity of androgen-regulated proteins and establish their hypothetical physiological role in the parasites. These analyses yield a possible explanation of how androgens exert their cysticidal effects through changes in the expression of proteins involved in cytoskeletal rearrangement, dynamic vesicular traffic and transduction of intracellular signals.

Taenia solium cysticercosis is a major public health problem in developing countries. Swine cysticercosis results in economic losses for pig farmers in disease endemic areas. Consumption of cysticercotic pork leads to taeniasis in humans. Eggs excreted in the faeces of T. solium carriers disseminate to humans and pigs through the faecal–oral route, thus maintaining the life cycle in endemic areas. An enzyme-linked immunoelectrotransfer blot (EITB) assay was developed using whole crude T. solium cysticercus antigens (WCA) for the diagnosis of swine cysticercosis. Sera from 30 swine with cysticercosis confirmed by magnetic resonance imaging were subjected to EITB assay. Sera from 50 swine that were raised in a government farm and not allowed to roam freely were included as negative controls. Two or more bands of 8, 11, 14, 24, 26 and 29 kDa were immunoreactive on blot with sera from all infected swine except two, and none from swine raised on the government farm. The overall sensitivity and specificity of EITB assay for diagnosis of swine cysticercosis were 93.3% and 100%, respectively. Hence, EITB assay based on WCA may be a suitable diagnostic tool for swine cysticercosis in endemic areas.

The yellow-necked mouse, Apodemus flavicollis, can be considered as a model for genetic polymorphism produced by the frequent presence of supernumerary or B chromosomes (Bs). Host genetic background is rarely taken into account in studies of parasite sex ratio. The main aim of this study was to investigate the range of infrapopulation sex ratios for nematode parasites of the yellow-necked mouse and to determine which factors most influence variation in parasite sex ratios. Six nematode species found in the collected yellow-necked mice were analysed. We confirmed the predominant pattern of female-biased sex ratios in vertebrate parasite infrapopulations. The presence of B chromosomes in host genomes played an important role in infrapopulations of Heligmosomoides polygyrus, Syphacia stroma and Trichuris muris, as hosts with B chromosomes carried a higher proportion of males. The relative increase of males in infrapopulations could result from a shift in parasite life history strategy, induced by adaptation to the specific host genotypes (Bs present). In a meta-analysis with previously published data, the sex determination system was demonstrated to play a significant role in nematode sex ratio variation, as well as specific life history patterns, such as the place of egg hatching.

Parasite distribution patterns in lotic catchments are driven by the combined influences of unidirectional water flow and the mobility of the most mobile host. However, the importance of such drivers in catchments dominated by lentic habitats are poorly understood. We examined parasite populations of Arctic charr Salvelinus alpinus from a series of linear-connected lakes in northern Norway to assess the generality of lotic-derived catchment-scale parasite assemblage patterns. Our results demonstrated that the abundance of most parasite taxa increased from the upper to lower catchment. Allogenic taxa (piscivorous birds as final host) were present throughout the entire catchment, whereas their autogenic counterparts (charr as final hosts) demonstrated restricted distributions, thus supporting the theory that the mobility of the most mobile host determines taxa-specific parasite distribution patterns. Overall, catchment-wide parasite abundance and distribution patterns in this lentic-dominated system were in accordance with those reported for lotic systems. Additionally, our study highlighted that upper catchment regions may be inadequate reservoirs to facilitate recolonization of parasite communities in the event of downstream environmental perturbations.

One hundred and sixty-one northern bobwhites (Colinus virginianus; hereafter ‘bobwhite’) were examined from the Rolling Plains ecoregion of Texas and western Oklahoma from 2011 to 2013. Complete necropsies yielded 13 species, of which two are new host (Gongylonema phasianella) and region (Eucoleus contortus) records and three (Dispharynx nasuta, Tetrameres pattersoni and Oxyspirura petrowi) are known to cause morbidity and mortality. Of the species found, Aulonocephalus pennula commonly occurred, Oxyspirura petrowi was intermediate in prevalence, and the remaining species were rare. Species richness was similar compared to studies from the southeastern U.S., but higher than studies from the same region. In addition, 12 of the 13 species were heteroxenous helminths, supporting the theory that heteroxenous helminths in semi-arid regions are more successful than monoxenous helminths. Prevalence and abundance of A. pennula and O. petrowi were higher in adult bobwhites than in juveniles. Abundance of A. pennula and O. petrowi was higher at southern locations compared to northern locations in the study area. Our study is the first to provide a current assessment of the bobwhite helminth community across the Rolling Plains ecoregion of the U.S.

There is a paucity of information on the diversity and occurrence of freshwater fish parasites in Australia. This study investigates the distribution and occurrence of a parasitic nematode of the genus Contracaecum in freshwater fish from south-eastern Australia. Fish (n = 508) belonging to nine species and eight families were collected from eight wetlands associated with the Murrumbidgee River floodplain in the southern Murray–Darling Basin and subjected to laboratory examination. Third-stage nematode larvae were found in eight of the nine fish species. The exception was the fly-specked hardeyhead (Craterocephalus stercusmuscarum), although only one specimen of this species was examined. Nematode larvae were identified as Contracaecum bancrofti using a combined morphological and molecular approach. The parasite was most prevalent in weather loach (Misgurnus anguillicaudatus; 34.8%) and Australian smelt (Retropinna semoni; 21.4%), followed by carp (Cyprinus carpio; 9.5%), of which the former and latter are non-native species. Contracaecum prevalence differed between locations, with Two Bridges having the highest number of infected fish. This may be due to the higher number of suitable host species collected at these localities. Contracaecum spp. are parasites with low host specificity that have also been reported in Australian marine fish, humans and piscivorous birds. The high parasite prevalence in the two abundant non-native fish species in the region suggests that they act as a suitable host for this endemic parasite, resulting in the increase in the parasite population. It would be interesting to study host–parasite interactions in this area, especially if introduced fish populations declined dramatically in the attempt to eradicate them or control their population.

We describe the larval developmental stages and life cycle of the dracunculid nematode Mexiconema cichlasomae in both the intermediate, Argulus yucatanus (Crustacea: Branchiura), and definitive hosts, Cichlasoma urophthalmus (Perciformes: Cichlidae), from the Celestun tropical coastal lagoon, Yucatan, Mexico. The morphological analyses showed significant differences between the total length of L1 found in M. cichlasomae gravid female and L2–L3 in A. yucatanus. This result indicates that the M. cichlasomae larval development occurs in the intermediate host. We obtained sequences from the small subunit (SSU) ribosomal marker from larval stages of M. cichlasomae in A. yucatanus and adult nematodes in C. urophthalmus. Our morphological and molecular results support conspecificity between M. cichlasomae larvae in A. yucatanus and the adult stages in C. urophthalmus. We briefly discuss the phylogenetic position of M. cichlasomae among the Daniconematidae, and provide evidence of the monophyly of the daniconematids associated with branchiurid intermediate hosts. Based on the phylogenetic results, we support the transfer of the Mexiconema genus to the family Skrjabillanidae and do not support the lowering of family Daniconematidae to subfamily.

Examination of rajid skates off the Algerian coast in the Mediterranean Sea revealed that three of the 33 Dipturus batis Linnaeus, 1758 examined harboured a new tapeworm species: Echeneibothrium algeriensis n. sp. This new species, collected from the anterior half of the spiral valves, is described on the basis of morphological data from light and scanning electron microscopy. The new species differs from previously described Echeneibothrium species by details of the scolex and loculi, total length, the length of the myzorhynchus, the number of proglottides, and the number of testes. Comparison of the diets of the ten skate species common in the Mediterranean basin indicates some varying degree of overlap, suggesting that host specificity in this host–parasite system is determined by other host and/or ecological variables such as adaptations of the parasites to their respective hosts, either on the morpho-anatomical level, in physiological characteristics of the parasite's habitat, in the trophic requirements for the successful transmission of the parasite, or in adaptations to the behavioural characteristics of the host. Furthermore, restricted overlap of E. algeriensis n. sp. with congeners in parasite assemblages of D. batis indicates some structuring according to attachment-site preferences. However, attachment-site preferences are not explained solely by morphological compatibility between bothridia and villi. This study reiterates the need to examine multiple factors synergistically in studies on host specificity of parasites, and the need to examine the parasite fauna of hosts across their entire geographical range in order to truly appreciate the biodiversity they harbour.

The golden snub-nosed monkey (Rhinopithecus roxellana) is an endangered species endemic to China. Relatively little is known about the taxonomic status of soil-transmitted helminths (STH) in these monkeys. Trichuris spp. (syn. Trichocephalus) are among the most important STHs, causing significant socio-economic losses and public health concerns. To date, five Trichuris species have been reported in golden monkeys, including a novel species, T. rhinopiptheroxella, based on morphology. In the present study, molecular and morphological analysis was conducted on adult Trichuris worms obtained from a dead golden snub-nosed monkey, to better understand their taxonomic status. Morphology indicated that the adult Trichuris worms were similar to T. rhinopiptheroxella. To further ascertain their phylogenetic position, the complete mitochondrial (mt) genome of these worms was sequenced and characterized. The mt genome of T. rhinopiptheroxella is 14,186 bp, encoding 37 genes. Phylogenetic analysis based on the concatenated amino acids of 12 protein-coding genes (with the exception of atp8) indicated that T. rhinopiptheroxella was genetically distinct and exhibited 27.5–27.8% genetic distance between T. rhinopiptheroxella and other Trichuris spp. Our results support T. rhinopiptheroxella as a valid Trichuris species and suggest that mt DNA could serve as a marker for future studies on the classification, evolution and molecular epidemiology of Trichuris spp. from golden snub-nosed monkeys.

Strongyloides fuelleborni is a soil-transmitted nematode parasite of non-human primates. The worm is prevalent also in human populations in Africa and South-East Asia. In this study, we amplified and sequenced a portion of the 18S ribosomal RNA gene (rRNA) and of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene of Strongyloides adult males recovered from faecal samples from long-tailed macaques (Macaca fascicularis) in Thailand and Lao PDR. The prevalence in Thailand was 31.1% (55/177) and in Lao PDR it was 62.1% (41/66), with an overall prevalence of 39.5% (96/243). All 18S rRNA sequences that we obtained (n = 96) showed 100% identity with published S. fuelleborni sequences. The 96 cox1 sequences that we obtained represented 32 new haplotypes. When included with the 17 previously known haplotypes from S. fuelleborni, the cox1 sequences fell into four clusters, which had clear geographical structure. This is the first molecular confirmation of S. fuelleborni in long-tailed macaques in Thailand and Lao PDR. Clearly, awareness needs to be raised of the zoonotic potential of S. fuelleborni. A monitoring programme should be organized, taking into account the role of reservoir hosts (i.e. monkeys) in the natural background of human strongyloidiasis caused by S. fuelleborni.

Despite the advances of molecular tools, new nematode species are still described mainly based on morphological characters. Parapharyngodon and Thelandros are two genera of oxyurids with unclear related taxonomic histories. Here we use morphological characters (linear measurements and categorical variables) and genetic information (18S rRNA, 28S rRNA and COI partial gene sequences) to confirm the relationships between representatives of these two genera and to determine whether they can be discriminated morphologically. Genetic results confirm the existence of two main clades, mostly congruent with Parapharyngodon and Thelandros genera but with several discordances. Thelandros is polyphyletic, with two of the species analysed (T. filiformis and T. tinerfensis) being part of the Thelandros clade, but with a third one (T. galloti) falling within the Parapharyngodon clade. Regarding the Parapharyngodon clade, P. cubensis, P. scleratus and Parapharyngodon sp. from Mexico form congruent lineages, while most P. echinatus samples cluster in another group, with one exception. Interestingly, P. micipsae samples are scattered across the Parapharyngodon clade, suggesting that they were misidentified or rather represent alternative morphotypes of other species. Morphological analysis identified the length of the tail, number of caudal papillae, position of the nerve ring, presence of caudal alae and length of the lateral alae as reliable characters to distinguish between Parapharyngodon and Thelandros genetic clades. Our study highlights the current taxonomic inconsistency in these groups, mainly derived from the exclusive use of morphological data. As such, we advocate for the routine implementation of molecular data in nematode taxonomic studies.

Diaphanocephalus galeatus collected from the small intestine of the lizard Dracaena paraguayensis in the Pantanal wetlands, State of Mato Grosso do Sul, Brazil, is redescribed. Genetic characterization and observations using scanning electron microscopy (SEM) were performed for the first time. The vouchers of D. galeatus and the type specimens of its congeners, deposited in the Coleção Helmintológica do Instituto Oswaldo Cruz (CHIOC), were consulted. Light and SEM observations revealed several undescribed features of D. galeatus, i.e. structure of the cephalic end and of the buccal capsule, position and morphology of deirids, presence of phasmids in females and presence of unpaired papilla on the membranous projection that covers the genital cone in males. After observation of the specimens deposited in the helminthological collection, D. jacuruxi is considered a synonym of D. galeatus, and D. diesingi, despite its incomplete description, is tentatively retained as valid due to the poor condition of the type material. The results also indicated low host specificity of D. galeatus, contradicting previous assertions. Genetic comparisons using patristic distances and phylogenetic trees generated from sequences of the 28S rRNA nuclear gene indicated that D. galeatus is closer to the taxa within Ancylostomatoidea and Strongyloidea than any lineage of Metastrongyloidea or Trichostrongyloidea. However, most of the nodal supports were low. Based on the genetic and morphological characterization, the validity of D. galeatus was confirmed. These data may serve for further comparative approaches for different populations of the parasite, from different hosts in different geographical areas, mitigating taxonomic confusions.

Diagnosis of cystic echinococcosis (CE) is at present mainly based on imaging techniques. Serology has a complementary role, partly due to the small number of standardized and commercially available assays. Therefore we examined the clinical performance of the SERION ELISA classic Echinococcus IgG test. Using 10 U/ml as a cut-off point, and serum samples from 50 CE patients and 105 healthy controls, the sensitivity and specificity were 98.0% and 96.2%, respectively. If patients with other infectious diseases were used as negative controls, the specificity decreased to 76.9%, which causes poor positive predictive values. However, if results between 10 and 15 U/ml are classified as indecisive, the specificity of positive results (≥15 U/ml) increased to 92.5% without greatly affecting the sensitivity (92.0%). Using this approach in combination with imaging studies, the SERION ELISA classic Echinococcosis IgG test can be a useful aid in the diagnosis of CE.

Neurocysticercosis is a parasitic disease caused by encysted larvae of Taenia solium in the human central nervous system. Cysts mainly affect the cerebral hemispheres, although they can also be found in ventricles, basal cisterns, and subarachnoid spaces, and rarely in the cerebellum. Given the impossibility of studying the disease in human patients, Cardona et al. (1999) developed a mouse model of neurocysticercosis, using Mesocestoides corti, a closely related cestode. This allows us to study the parasite–host relationship and the mechanisms involved in the disease, in order to improve the therapy. In this murine model of neurocysticercosis, the location of tetrathyridia in parenchyma, ventricles and meninges has already been reported. The aim of this work is to report the cerebellum as a new location for M. corti tetrathyridia in the murine model of neurocysticercosis. A murine model that reproduces the human pathology is essential to evaluate the symptomatology and response to drug treatment in experimentally infected mice.

The parasite Cyathostomum pateratum, which occurs in the large intestine of equines, is a common species of the subfamily Cyathostominae. Cyathostominae nematodes are a complex nematode group for which only limited genetic information has been reported. To re-examine the phylogenetic relationships among Cyathostominae nematodes, we sequenced the complete mitochondrial (mt) genome of Cy. pateratum and compared it with the mt genome of the congeneric species Cyathostomum catinatum. The complete mtDNA sequence of Cy. pateratum was 13,822 bp in length, 16 bp shorter than that of Cy. catinatum. The mtDNA sequences of both species contained 12 protein-coding genes, two rRNA genes and 22 tRNA genes, and all 36 genes were transcribed in the same direction and in the same strand. Pairwise comparisons of the 12 predicted amino acid sequences between Cy. catinatum and Cy. pateratum revealed differences of 0.4–3.1%; the least conserved sequence was that of cytochrome c oxidase subunit 3 (cox3). Phylogenetic analysis of the concatenated amino acid sequences using Bayesian inference and maximum likelihood methods showed that Cy. catinatum and Cy. pateratum clustered together with very high nodal support, and Cylicostephanus goldi was closer to the Cyathostomum nematodes than to other Cyathostominae nematodes. The mtDNA sequence of Cy. pateratum is reported here for the first time. The study will shed some light on the genetic evolution among parasitic nematodes in Cyathostomum.