The potential role of roaming dogs in establishing a geographically novel life cycle of taeniids (Echinococcus spp. and Taenia spp.) in a non-endemic area.

INTRODUCTION: Cystic Echinococcosis (CE) is endemic in humans and livestock in many pastoral communities in Kenya. The distribution of the disease is enhanced by several factors, including livestock trade, which has allowed for the spread of CE to non-endemic areas such as western Kenya. Dogs' roaming behaviour, with consequent contamination of the environment with intestinal parasites, could then lead to parasite establishment. This study examined dogs' infection levels with taeniid eggs and their potential role in contaminating the environment with intestinal parasites. METHODOLOGY: We selected sixteen ruminant slaughterhouses in Busia and Bungoma Counties, and around each slaughterhouse we identified ten homesteads owning free-roaming dogs. We administered a questionnaire on dog management practices to the homestead owner and collected a faecal sample from the dog's rectum. In homesteads around 8 of the 16 slaughterhouses, we collared dogs with a GPS tracker to assess their movement patterns. The faecal samples were examined microscopically following zinc-chloride sieving-floatation technique for the presence of taeniid eggs and other canine intestinal parasites. Polymerase Chain Reaction - Restriction Fragment Length Polymorphism of NADH dehydrogenase subunit 1 gene and sequencing were used to confirm taeniid eggs identified during microscopy. Additionally, the Coproantigen-ELISA was used to detect the presence of taeniid antigen in a sub-set of the faecal samples. RESULTS: Helminths detected in the 155 dogs sampled included hookworms (n = 92; 59.4%), ascarids (n = 15; 9.7%), and taeniids (n = 1; 0.6%). Through Copro-PCR, 13 eggs extracted from the sample of the only taeniid infected dog were sequenced and identified as E. canadensis (G6/7) [n = 1], Taenia multiceps [n = 1], and Taenia serialis [n = 6]; the remaining were indeterminate. Of the 77 faecal samples tested for E. granulosus sensu lato (s. l.) with the Copro-ELISA, 64 (83.1%) were negative, 12 (15.6%) were positive, while 1 (1.3%) was suspicious. The dogs travelled a median of 13.5 km daily, and 28 dogs visited the slaughterhouses during the 5-day recording period. CONCLUSION: The results indicate a relatively high carriage of zoonotic parasites by free-roaming domestic dogs in western Kenya, which poses a risk to human and livestock populations. We report for the first time a domestic lifecycle of Echinococcus canadensis and Taenia multiceps in western Kenya, as well as a presumptive sylvatic cycle of coenurosis by T. serialis. We recommend an extensive and ongoing Copro-antigen survey of dog faeces, broader assessment of dog parasites with zoonotic potential, adherence to slaughterhouse management practices, and dog-ownership programmes to highlight the importance of deworming and restricted dog movements.

Comparison of multiplex copro PCR with coproscopy followed by PCR on recovered eggs for the detection of Echinococcus granulosus and Taenia spp. infection in dogs.

Echinococcosis and taeniasis are important helminth diseases that carry considerable impact on human and animal health. Domestic dogs and other canids are definitive hosts for several parasites of this group, including Echinococcus granulosus, Taenia multiceps, T. ovis, T. hydatigena and E. multilocularis. Detection of infection in dog populations is imperative for estimating the risk to susceptible humans and animals, and for its mitigation through prevention measures in dogs, other animals and humans. To date, identification of taeniid eggs, antigens or DNA in fecal samples are the most practical diagnostic modalities available for canine definitive hosts. Although widely used for this purpose, there is limited information comparing copro PCR and combined coproscopy-PCR protocols for the detection of taeniids. In the current study, a widely used multiplex PCR was performed on a large number of dog fecal samples using DNA extracted directly from feces. The samples were also tested by fecal flotation and coproscopy, eggs were isolated from microscopically-positive samples and extracted DNA was tested using the same multiplex PCR. The total number of taeniid positive samples detected using both methods was 46/317 (14.5%), including 10/317 (3.2%) E. granulosus positive samples. Both copro PCR and coproscopy have identified an equal number of samples as taeniid positive (n = 32). However, for the purpose of identification to species level, the copro PCR was significantly more sensitive than coproscopy followed by PCR on isolated eggs (sensitivity 0.7 vs. 0.41, p = 0.012), with 32/317 (10.1%) and 19/317 (6%) positive samples identified, respectively. The difference in identification of E. granulosus was highly apparent, as the majority of the E. granulosus positive samples (8/10) were detected by the copro PCR only. Coproscopy and egg PCR have identified 5/317 (1.6%) positive samples not detected by the copro PCR, including only a single sample (0.3%) positive for E. granulosus. Adding these positive samples to those identified by the copro PCR did not significantly improve the overall sensitivity (p = 0.074). Therefore, using both copro PCR and coproscopy in parallel may not be advantageous for taeniid detection and identification, at least until the egg PCR is further optimized and performs better. These results should be weighed against the different advantages that coproscopy based approach may offer.

A rare case of human taeniasis caused by Taenia saginata with species undetermined cysticercosis.

Taeniasis and cysticercosis, which are caused by Taenia saginata, Taenia solium and Taenia asiatica, are zoonotic parasitic infections with a significant disease burden worldwide. There is consensus amongst experts that T. saginata is a common tapeworm that causes taeniasis in humans as opposed to cysticercosis. This case study of a middle-aged Tibetan man conducted in 2021 challenges the prevailing notion that T. saginata exclusively causes taeniasis and not cysticercosis by documenting symptoms and laboratory studies related to both taeniasis and multiple cysticercosis. The patient's medical record with the symptoms of taeniasis and cysticercosis was reviewed, and the tapeworm's proglottids and cyst were identified from the patient by morphological evaluation, DNA amplification and sequencing. The patient frequently experienced severe headaches and vomiting. Both routine blood screenings and testing for antibodies against the most common parasites were normal. After anthelmintic treatment, an adult tapeworm was found in feces, and medical imaging examinations suggested multiple focal nodules in the brain and muscles of the patient. The morphological and molecular diagnosis of the proglottids revealed the Cestoda was T. saginata. Despite the challenges presented by the cyst's morphology, the molecular analysis suggested that it was most likely T. saginata. This case study suggests that T. saginata infection in humans has the potential to cause human cysticercosis. However, such a conclusion needs to be vetted by accurate genome-wide analysis in patients with T. saginata taeniasis associated with cysts. Such studies shall provide new insights into the pathogenicity of T. saginata.

Urinary neopterin reflects immunological variation associated with age, helminth parasitism, and the microbiome in a wild primate.

Neopterin, a product of activated white blood cells, is a marker of nonspecific inflammation that can capture variation in immune investment or disease-related immune activity and can be collected noninvasively in urine. Mounting studies in wildlife point to lifetime patterns in neopterin related to immune development, aging, and certain diseases, but rarely are studies able to assess whether neopterin can capture multiple concurrent dimensions of health and disease in a single system. We assessed the relationship between urinary neopterin stored on filter paper and multiple metrics of health and disease in wild geladas (Theropithecus gelada), primates endemic to the Ethiopian highlands. We tested whether neopterin captures age-related variation in inflammation arising from developing immunity in infancy and chronic inflammation in old age, inflammation related to intramuscular tapeworm infection, helminth-induced anti-inflammatory immunomodulation, and perturbations in the gastrointestinal microbiome. We found that neopterin had a U-shaped relationship with age, no association with larval tapeworm infection, a negative relationship with metrics related to gastrointestinal helminth infection, and a negative relationship with microbial diversity. Together with growing research on neopterin and specific diseases, our results demonstrate that urinary neopterin can be a powerful tool for assessing multiple dimensions of health and disease in wildlife.

The prevalence of Taenia spp. in pigs slaughtered in Kinshasa.

Taenia hydatigena is a non-zoonotic worm that has dogs and wild canids as definitive hosts. Its presence induces cross reactions in certain diagnostic tests for porcine cysticercosis caused by T. solium, the occurrence of which has a considerable public health and economic impact. In the Democratic Republic of the Congo (DR Congo), T. solium is considered endemic, however, the prevalence of T. hydatigena has not been estimated yet. The objective of the study was therefore to estimate the prevalence of T. hydatigena cysticercosis by serological and molecular diagnostic tools in pigs slaughtered in DR Congo. A total of 480 pigs slaughtered in 6 slaughter slabs in Kinshasa, DR Congo, were examined. The thoracal and abdominal cavity organs were inspected for cysts, which were analyzed using PCR-RFLP. Furthermore, 480 sera were collected, and analyzed for the presence of circulating Taenia spp. cysticercus antigens, using the B158/B60 Ag-ELISA. Upon inspection of the carcass, 41 cysts suspected to be metacestodes of Taenia spp. were collected, from the following viscera: spleen (24/41, 59%), liver (13/41, 32%), intestine (3/41, 7%) and lung (1/41, 2%). Molecular analyses revealed a T. hydatigena prevalence of 0.2% (95% CI: 0.0001-0.0116), based on a single lesion (1/480), taken from the spleen. Out of the 480 sera collected, the presence of circulating Taenia spp. cysticerci antigens was detected in 32 (6.7%; 95% CI: 4.5-11.2). The results of this study revealed that T. hydatigena is present in pigs sold in markets in the city of Kinshasa in DR Congo, albeit at a very low prevalence, thus the impact on the interpretation of the B158/B60 seems low in this setting. Detection of circulating antigens in porcine sera by Ag-ELISA, shows that pigs slaughtered in Kinshasa, DR Congo, were infected with viable cysticerci of Taenia spp. which in turn can infect humans.

Decreased embryo implantation in rabbits infected with Taenia pisiformis.

Parasitic infections can have detrimental effects on the reproductive capacity of their hosts. Infections by the cestode Taenia pisiformis in rabbits is generally not associated with increased mortality of offspring or with loss of maternal body condition but can result in reduced fecundity, and increased circulating progesterone levels have been reported in infected does compared to uninfected ones. In the present study, the possibility that T. pisiformis infection affects fecundity by reducing embryonic implantation was examined. Seven anesthetized New Zealand does were orally infected with 1000 eggs of T. pisiformis, and seven were administered saline. Seven weeks after infection, does were mated, and 7 days after, humanely sacrificed. A decrease in the number of implanted embryos and a decrease in the size of the embryo vesicles in infected does were observed (P ≤ 0.02, Student's t-test). There was a negative correlation between the number of hepatic granulomas and embryo implantation (ρ = - 0.8, P = 0.04, Spearman's test).

Taenia hydatigena larvae vesicular concentrates increase Anti-OVA IgG and the production of some cytokines in rats.

The effects of administration of four different fractions of T. hydatigena larvae vesicular concentrate (ThLVC) prior to immunization with ovalbumin (OVA) in rats on different parameters of the immune response were evaluated. The amount of anti-OVA IgG by ELISA, amount of blood eosinophils (BE), percentage of cell subpopulations by flow cytometry (CD3+/CD4+, CD3+/CD8+, CD3-/CD45RA+, and CD11b/c+), and production of serum cytokines by bead-based immunoassays (IL-2, IL-4, INFγ, IL-5, TNFα, GM-CSF, IL-17F, IL-17A IL-13, IL-22, and IL-6) were measured. Rats receiving total-ThLVC (p ≤ 0.05) and fraction ThLVC30-100 kDa (p < 0.001) prior to OVA administration produced higher amounts of anti-OVA IgG than rats receiving OVA alone. Rats that were only administered with OVA showed a strong increase in BE that was significantly correlated (r = 0.72, p < 0.001) with an increase in IL-5 in the blood. However, rats that received any of the ThLVC fractions prior to administration of OVA did not show these increases. In general, administration of ThLVC30-100 kDa prior to administration of OVA increased (p < 0.05) the percentage of B, CD4, and CD8 lymphocytes in the spleen and mesenteric lymph nodes. Rats that received ThLVC total fraction and OVA showed an increase (p < 0.05) in IL-2, IL17F, and IL22. The results of this study show that total-ThLVC and ThLVC30-100 kDa modify the immune response of rats in differentiated ways. Our observations suggests that both fractions of ThLVC have the potential to be used as adjuvants.

Identification and expression analysis of a new small ubiquitin-like modifier from Taenia pisiformis.

The small ubiquitin-like modifier (SUMO) plays important roles, with the SUMOylation pathway as one of its core components. In the present work, a single SUMO gene was initially identified from Taenia pisiformis and designated as TpSUMO. Bioinformatic analysis showed that the TpSUMO gene contained a 309 bp open reading frame (ORF), encoding 102 amino acids, and had a predicted molecular weight of ∼12 kDa. The amino acid sequence of TpSUMO was deduced and it shared 44.00% identity with human SUMO2 (HsSUMO2) and exhibited more than 97.78% identity with SUMOs from Taenia and Echinococcus. TpSUMO possessed a putative non-consensus site (FK11MG) within its N-terminus and a typical di-glycine (GG) motif at the C-terminus. Basic local alignment search tool (BLAST) analysis showed that only a single SUMO-related ortholog was present in each set of known genome data for fourteen tapeworm species. The precursor His-TpSUMO-FL, mature His-TpSUMO-GG and mutant His-TpSUMO-GGK11R proteins (∼18 kDa) were expressed in Escherichia coli Rosseta (DE3), and rabbit polyclonal anti-TpSUMO was generated with a high titer of 1.28 × 105. In vitro SUMOylation assay results showed that TpSUMO multimer formation in the His-TpSUMO-GG reaction could be catalyzed by the human SAE1/SAE2 and UBC9 conjugation system, but K11R mutation disrupted TpSUMO chain synthesis. Quantitative real-time PCR (qRT-PCR) further revealed that TpSUMO was ubiquitously expressed in different stages of T. pisiformis and in higher levels during an early development phase (day 14) of adult worms. Immunofluorescence localization showed that TpSUMO was detected in the bladder wall of cysticerci, in the testis in immature segment, and within eggs in the gravid proglottids. These findings indicated that TpSUMO is a new member of the SUMO protein family and may play a vital role in regulation of functions within proteins involved in worm growth and development.

In vitro system for the growth and asexual multiplication of Taenia crassiceps cysticerci.

Taenia solium is the aetiological agent of cysticercosis, a zoonosis that causes severe health and economic losses across Latin America, Africa and Asia. The most serious manifestation of the disease is neurocysticercosis, which occurs when the larval stage (cysticercus) establishes in the central nervous system. Using Taenia crassiceps as an experimental model organism for the study of cysticercosis, we aimed to identify the in vitro conditions necessary to allow parasite development at the short- and long terms. First, cysticerci were incubated for 15 days in different media and parasite densities. The number of buddings and cysticerci diameter were measured to evaluate asexual multiplication and parasite growth, respectively. Vitality was determined by trypan blue staining and morphology analysis. As a result, high cysticerci density and medium containing FBS and the excretion/secretion (E/S) products of feeder cells induced parasite survival, growth and multiplication. Then, the long-term (5 weeks) incubation of the parasites in co-culture with feeder cells was evaluated. Consequently, the mammalian cell lines induced a significant increase in total parasite volume while axenic cultures did not show any statistically significant change over time. In this study, the proper conditions to maintain T. crassiceps in vitro are described for the first time in a simpler and more controlled setting other than experimental infections. In addition, it was shown that cysticerci growth, survival and asexual multiplication depend on a complex network of secreted factors from both parasite and host.

Comparison of mitochondrial genetic variation of Taenia hydatigena cysticerci from China and Mongolia.

Parasitic infection is one of the many challenges facing livestock production globally. Cysticercosis tenuicollis is a common parasitic disease in domestic and wild ruminants (intermediate host) caused by the larval stage of Taenia hydatigena that primarily infects dogs (definitive host). Although genetic studies on this parasite exist, only a few describe the genetic variation of this parasite in Mongolia. Our aim was thus, to identify the mitochondrial differences in ovine isolates of Cysticercus tenuicollis entering China from Mongolia and comparison with existing Chinese isolates from sheep and goats based on the recently described PCR-RFLP method and mitochondrial genes of NADH dehydrogenase subunit 4 (nad4) and the NADH dehydrogenase subunit 5 (nad5). Sixty-nine isolates were collected during routine veterinary meat inspections from sheep that originated from Mongolia, at the modern slaughterhouses in Erenhot City, Inner Mongolia. Additional 114 cysticerci were also retrieved from sheep and goats from northern (Inner Mongolia Autonomous Region, Ningxia Hui Autonomous Region, and Gansu Province), western (Tibet Autonomous Region), and southern (Jiangxi Province and Guangxi Province) China. The PCR-RFLP approach of the nad5 showed nine mitochondrial subclusters A1, A2, A3, A5, A8, A9, A10, A11, and B of T. hydatigena isolates from sheep and goats from Mongolia and China. Meanwhile, haplogroup A1 RFLP profile was more widespread than other variants. These data supplements existing information on the molecular epidemiology of T. hydatigena in China and Mongolia and demonstrate the occurrence of similar genetic population structures in both countries.

Global epidemiology and molecular biology of Taenia multiceps: a comparative meta-analysis and in silico analysis study.

In the present study, all published data on the epidemiology and molecular characters of Taenia multiceps were systematically collected from relevant databases (e.g. PubMed, Scopus, National Center for Biotechnology Information), and combined in various statistical and genetic analyses as a contribution to a better understanding of the epidemiology of this ubiquitous taeniid worldwide. While 5.8% of the key hosts (dogs) from various countries had T. multiceps, grey wolves displayed the highest prevalence (21.6%) among the definitive hosts. Small ruminants are the main intermediate hosts and carry the coenuri in various locations, but most commonly in the central nervous system (CNS). Cerebral coenuri were confirmed in 53% of sheep exhibiting neurological symptoms, and infected animals often had only a single coenurus in the brain. Sheep had a higher prevalence (8.8%) of CNS coenuri than goats (5.8%); however, extra-CNS coenuri were detected more frequently in goats than in sheep. In either case, the difference between sheep and goats was statistically insignificant. Analysis of 233 partial cytochrome oxidase subunit I nucleotide sections for T. multiceps revealed high haplotype and low nucleotide diversities. Fifty-one haplotypes were detected circulating in 6 geographic populations. China, Iran and Turkey had 2 major haplotypes, whereas Italy and Egypt shared 3. Haplotypes from Greece circulate worldwide, and displayed similar gene flow values when compared with the other populations. There were no distinct patterns for haplotype distribution in relation to the infected hosts or coenuri locations. The existence of genetic variants in T. multiceps was highlighted, but needs further studies.

Evaluating the effect of curcumin on the metacestode of Taenia crassiceps.

Curcumin, a curcuminoid present in the rhizome of the plant Curcuma longa has multiple pharmacological effects including anticarcinogenic and anti-inflammatory properties. This work evaluates the anthelmintic effect of the curcumin molecule (98% pure) on Taenia crassiceps cysticerci viability in vitro. Cysticerci incubated in the presence of increasing concentrations of curcumin showed a dose-dependent mortality correlated with a significant increase in the production of reactive oxygen species and a partial inhibition of thioredoxin-glutathione reductase, the only disulfide reductase present in these parasites. At 500 µM curcumin, a 100% of cysticerci lethality was obtained after 2 h of treatment. These results suggest the curcumin-induced oxidative stress could be in the origin of the anthelminthic effect of curcumin. Mice with cysticerci were injected intraperitoneally with 20, 40, or 60 mM curcumin daily for 30 days. A decrease in the burden of cysticerci (46%) was observed with a 60 mM dose of curcumin, supporting this compound as a potential anthelmintic drug.

Experimental animal models and their use in understanding cysticercosis: A systematic review.

BACKGROUND: Cysticercosis and Neurocysticercosis (NCC) can be studied using several animal species in experimental models which contributes to the understanding of the human form of the disease. Experimental infections of Taenia spp. are vital in explaining the modes of transmission of the parasite and helps the understanding of transmission of the parasite in humans and thus may be useful in designing therapeutic and immune-prophylactic studies to combat the disease. Thus, this systematic review aims to explore the existing experimental animal models to the understanding of cysticercosis in both humans and animals and elucidate the risk factors of cysticercosis and identify the Taenia spp. used in these models. METHODOLOGY: We systematically identified all publications from the Web of Science, Google Scholar, and Pubmed regarding experimental animal models using Taenia spp. that cause cysticercosis in both humans and animals. 58 studies were identified for eligibility. Of these, only 48 studies met the inclusion criteria from which data extraction was done and presented descriptively. RESULTS: Pigs, cattle, gerbils, mice, rats, voles, monkeys, cats, dogs, and goats were used in which T. solium, T. saginata, T. saginata asiatica, T. crassiceps and T. asiatica were studied. The routes used to induce disease were; oral, intravenous, subcutaneous, intramuscular, intraperitoneal, intraarterial, intracranial, intraduodenal, and surgical routes using eggs, oncospheres, and proglottids. Besides, the establishment of infection using eggs and oncospheres was affected by the route used to induce infection in the experimental animals. The cysticerci recovery rate in all the experimental studies was low and the number of animals used in these experiments varied from 1 to 84. Although not analysed statistically, sex, age, and breed of animals influenced the cysticerci recovery rate. Additionally, the cysticerci recovery rate and antibody-antigen levels were shown to increase with an increase in the dose of oncospheres and eggs inoculated in the animals. Contrasting results were reported in which the cysticerci recovery rate decreased with an increase in the dose of eggs inoculated. CONCLUSION: This review describes the various animal experiments using Taenia species that cause cysticercosis highlighting the animals used, age and their breed, the routes of infection used to induce disease and the sample size used, and the cysticerci recovery rate in these animal models.

Genomic palaeoparasitology traced the occurrence of Taenia asiatica in ancient Iran (Sassanid Empire, 2th cent. CE-6th cent. CE).

Palaeoparasitology investigates parasitological infections in animals and humans of past distance by examining biological remains. Palaeofaeces (or coprolites) are biological remains that provide valuable information on the disease, diet, and population movements in ancient times. Today, advances in detecting ancient DNA have cast light on dark corners that microscopy could never reach. The archaeological site of the Chehrabad salt mine of Achaemenid (550-330 BC) and Sassanid (third-seventh century AD) provides remains of various biotic and abiotic samples, including animal coprolites, for multidisciplinary studies. In the present work, we investigated coprolites for helminth eggs and larvae by microscopy and traced their biological agents' DNA by Next Generation Sequencing. Our results revealed various helminths, including Taenia asiatica, the species introduced in the 1990s. Implementing advanced modern molecular techniques like NGS gives a paramount view of pathogenic agents in space and time.

Increased iron uptake in the bladder wall of racemose cysts of Taenia solium.

Racemose neurocysticercosis is an aggressive infection caused by the aberrant expansion and proliferation of the bladder wall of the Taenia solium cyst within the subarachnoid spaces of the human brain. The parasite develops and proliferates in a microenvironment with low concentrations of growth factors and micronutrients compared to serum. Iron is important for essential biological processes, but its requirement for racemose cyst viability and proliferation has not been studied. The presence of iron in the bladder wall of racemose and normal univesicular T. solium cysts was determined using Prussian blue staining. Iron deposits were readily detected in the bladder wall of racemose cysts but were not detectable in the bladder wall of univesicular cysts. Consistent with this finding, the genes for two iron-binding proteins (ferritin and melanotransferrin) and ribonucleotide reductase were markedly overexpressed in the racemose cyst compared to univesicular cysts. The presence of iron in the bladder wall of racemose cysts may be due to its increased metabolic rate due to proliferation.

A chromosome-level genome assembly for the rabbit tapeworm Taenia pisiformis.

Taenia pisiformis is one of the most widespread gastrointestinal parasites and its larvae (cysticercosis) causes significant economic loss to rabbit industry. No efficient drug is available for this disease to date. To better understand its genomics, we assembled a 211-Mb high quality genome of T. pisiformis at chromosome level with a scaffold N50 size of 20 Mbp. Totally, 12,097 protein-coding genes was predicted from the genome. Genome-level phylogenetic analysis confirmed the taxonomic affiliations with other tapeworms and revealed that T. pisiformis diverged from its closely related relative T. hydatigena âˆ¼ 14.6 Mya. Comparative genomic analyses revealed that the T. pisiformis genome was characterized by adaptive features of strong positive selection signals from carbohydrate/lipid metabolism and body surface integrity, and of expanded gene families related to metabolism of amino acids and lipids. The high-quality genome of T. pisiformis constitutes a resource for the comparative genomics and for further applications in general parasitology.

Oxfendazole induces protein catabolism and gluconeogenesis in experimental neurocysticercosis.

Neurocysticercosis (NCC) is an endemic public health disease of the central nervous system highly related to epilepsy and seizures. Taenia crassiceps is an experimental model used for NCC and biochemical studies of the host-parasite relationship. For the past 50 years the NCC therapeutic treatment is performed with albendazole (ABZ) and praziquantel which opens a gap for new therapies due to parasitic resistance and other adverse effects of the drugs. Oxfendazole (OXF) is an albendazole derivative with efficacy against tissue cestodes of veterinary importance. The aim of this study was to determine the metabolic impact of OXF on T. crassiceps cysticerci intracranially inoculated in Balb/C mice. The animals were intracranially inoculated with T. crassiceps cysticerci and 30 days later received single dose oral treatment of OXF, ABZ and NaCl 0.9% (control group). The metabolic impact was quantified through the detection of metabolites from glycolysis, anaerobic fermentation of lactate and propionate, tricarboxylic acid cycle, protein catabolism, fatty acids oxidation. The differences observed in the concentrations of metabolites from the OXF treated group showed that the drug induced gluconeogenesis, increase in protein catabolism, fatty acids oxidation and propionate fermentation in comparison to the ABZ and control treated groups. In conclusion, OXF induced greater metabolic impact in T. crassiceps cysticerci than the standard NCC treatment, ABZ, showing that it may represent an alternative drug for its treatment.

A novel use of a geometric morphometric technique to distinguish human parasite eggs of twelve different species.

Copro-microscopic diagnostic methods are the most common approach for screening patients with parasitic infections. However, expertise is required to identify helminthic eggs from fecal specimens. Consequently, new methods are required to support accurate species identification. Novel technologies have recently been developed for the classification of organisms, including geometric morphometric (GM) approaches. In this study, the outline-based GM approach was used to distinguish the eggs of 12 common human parasite species, including Ascaris lumbricoides, Trichuris trichiura, Enterobius vermicularis, hookworm, Capillaria philippinensis, Opisthorchis spp., Fasciola spp., Paragonimus spp., Schistosoma mekongi, Taenia spp., Hymenolepis diminuta and Hymenolepis nana. The GM analysis revealed that the size cannot be used as the main variable in the identification of parasite species at the egg stage, producing only 30.18% overall accuracy. However, comparisons of shape based on the Mahalanobis distances between pairs of parasite species showed significant differences in all pairs (p < 0.05). The shape analysis produced 84.29% overall accuracy. This is the first time that outline-based GM has been preliminarily confirmed as a valuable approach to support copro-microscopic analysis, in order to effectively screen helminth eggs. However, further studies with a larger set of helminth eggs and artefacts should be carried out to increase confidence in the identification of parasite species in the absence of local experts.

Prevalence and phylogeography of Taenia hydatigena metacestodes from goats of India.

The study determined the prevalence and genetic population structure relationships of Cysticercus tenuicollis (Taenia hydatigena metacestode) retrieved from the goats slaughtered in north India. An overall prevalence of 9.62% (59/613) was recorded. Genetic population structure relationships were assessed by targeting partial cytochrome c oxidase subunit 1 mitochondrial gene sequence. Phylogenetic tree analysis revealed that all the present study representative isolates (n = 7) formed a major clade and grouped with T. hydatigena isolates retrieved from sheep, goats, pigs and dogs, originating from China, Iran, Nigeria, Ghana and Poland. However, a single isolate from Himachal Pradesh (isolate 3) formed a subgroup within the clade. The neutrality and diversity indices revealed high values of haplotype diversity [Hd = 0.99695 (0.95238­1.0000)] and low nucleotide diversity (π = 0.49276), which was indicative of demographic expansion and low gene flow, suggesting that Indian T. hydatigena isolates were not genetically differentiated. Tajima's D (−1.26988) and Fu and Li's D statistics values (−0.74556) were negative, demonstrating deviations from neutrality and both propounded recent population expansion or purifying selection. Results highlighted a low genetic diversity of T. hydatigena metacestodes across the geographical range of north India.

In vitro metabolic stress induced by nitazoxanide and flubendazole combination in Taenia crassiceps cysticerci.

Taenia crassiceps is often used as experimental model for T. solium cysticercosis studies. Currently cysticercosis antiparasitic treatment is based on albendazole and praziquantel which may present side effects and parasitic resistance. The search for other antiparasitic drugs is necessary. Nitazoxanide (NTZ) and flubendazole (FLB) are broad spectrum antiparasitic drugs that present anti-cysticercosis effect. Metabolic analyses help to determine the impact of these drugs on parasites. The aim of this study was to determine the impact on the production and excretion of organic metabolites in T. crassiceps cysticerci after in vitro exposure to NTZ and FLB, isolated or in combination. T. crassiceps cysticerci were culture in RPMI medium and exposed to 10 µg/mL of NTZ, 10 µg/mL of FLB or 10 µg/mL of NTZ +10 µg/mL of FLB. 24 h after exposure, the parasites were chromatographic analyzed to determine the impact of these drugs on glycolysis, homolactic fermentation, tricarboxylic acid cycle, fatty acids oxidation and proteins catabolism. It was possible to determine that the drugs combination induced greater metabolic impact on cysticerci in comparison to the isolated drugs exposure. The drugs combination induced gluconeogenesis, metabolic acidosis, increase in tricarboxylic acid cycle and in proteins catabolism. While the NTZ isolated exposure induced metabolic acidosis and protein catabolism and the FLB isolate exposure induced gluconeogenesis and protein catabolism. These results show that the combination of drugs with different modes of action increase the antiparasitic effect and may be indicated as alternative cysticercosis treatments.