Cestode Research Articles

Experience of eradicating parasites of laboratory rats in conventional vivarium

The purpose of the research is to test different dehelminthization schemes of laboratory rats infected with cestodes Rodentolepis nana and nematodes Syphacia muris and evaluate the significance of combined environment disinfection measures. The practical experience of eradication (helminth eradication) in animals in a conventional vivarium was described.Materials and methods. Experiments were conducted to study the efficacy of anthelmintics and administration schemes against cestode and nematode infections in laboratory rats. In the first experiment, praziquantel was used at a dose of 10 mg/kg to treat rats infected with R. nana. In the second experiment, the comparative efficacy of fenbendazole, albendazole, and pyrantel was evaluated against syphaciosis at the recommended dosages of 20, 10, and 12.5 mg/kg, respectively. Each drug was given orally, individually, twice with an interval of 7 days. The third experiment tested different schemes for treating syphaciosis with fenbendazole. One group of rats was given the drug orally individually using an esophageal tube at a dose of 20 mg/kg once a day for 7 consecutive days. Other groups were given fenbendazole daily with food for 7 days (150 mg fenbendazole per 1 kg of food). In all three experiments, all animals were divided into groups, and their cells underwent a complex of additional disinfection measures, and those kept in cages without disinfection.Results and discussion. Praziquantel showed 100% efficacy at a single dose of 10 mg/kg in R. nana therapy. In animals without additional disinfection procedures, cestode eggs were again recorded starting from day 14 after the drug administration. In the group of animals with disinfection measures, pathogens were not detected during the experiment. Double administration of fenbendazole, albendazole and pyrantel in the recommended dosages against syphaciosis did not result in eradicated nematodes in the animals. The disinfection did not affect the obtained results. Fenbendazole administered daily for 7 days ensured helminth eradication in animals. However, on day 7 after the therapy, Syphacia sp. eggs were again found in the groups that received the drug individually intragastrically through a tube, regardless of whether their cells were disinfected. The animals that received fenbendazole with food and were regularly disinfested remained free from nematodes throughout the experiment until the additional disinfection measures were cancelled. In the absence of disinfection, released helminth eggs were recorded on day 14 after therapy.

An Initial Study on the Use of Machine Learning and Radio Frequency Identification Data for Predicting Health Outcomes in Free-Range Laying Hens.

Maintaining the health and welfare of laying hens is key to achieving peak productivity and has become significant for assuring consumer confidence in the industry. Free-range egg production systems represent diverse environments, with a range of challenges that undermine flock performance not experienced in more conventional production systems. These challenges can include increased exposure to parasites and bacterial or viral infection, along with injuries and plumage damage resulting from increased freedom of movement and interaction with flock-mates. The ability to forecast the incidence of these health challenges across the production lifecycle for individual laying hens could result in an opportunity to make significant economic savings. By delivering the opportunity to reduce mortality rates and increase egg laying rates, the implementation of flock monitoring systems can be a viable solution. This study investigates the use of Radio Frequency Identification technologies (RFID) and machine learning to identify production system usage patterns and to forecast the health status for individual hens. Analysis of the underpinning data is presented that focuses on identifying correlations and structure that are significant for explaining the performance of predictive models that are trained on these challenging, highly unbalanced, datasets. A machine learning workflow was developed that incorporates data resampling to overcome the dataset imbalance and the identification/refinement of important data features. The results demonstrate promising performance, with an average 28% of Spotty Liver Disease, 33% round worm, and 33% of tape worm infections correctly predicted at the end of production. The analysis showed that monitoring hens during the early stages of egg production shows similar performance to models trained with data obtained at later periods of egg production. Future work could improve on these initial predictions by incorporating additional data streams to create a more complete view of flock health.

Inhibition of mouse colon cancer growth following immunotherapy with a fraction of hydatid cyst fluid

BackgroundHydatid cyst is the larval stage of the tape worm Echinococcus granulosus which is located in human and livestock viscera. There are some scientific evidences indicating that parasitic infections induce antitumor activity against certain types of cancers. In this study, the effects of a fraction of hydatid cyst fluid on colon cancer tumor in BALB/c mice were investigated. Materials and methodsIn this experimental work six groups of mice were challenged with mouse colon cancer cells. 5 days later when the sign of tumor growth in mice was seen, group 1–4 were injected with hydatid cyst fluid, the 78 kDa fraction, live protoscolices and BCG respectively. Group five was injected with alum alone and the sixth group left intact without any injection. The size of the tumor was measured and compared in all groups. Then blood samples of mice were evaluated for serum cytokine levels. ResultIn mice injected with hydatid cyst antigens especially a fraction of hydatid cyst fluid, tumor size was smaller than the that of control groups and the difference of tumor size in cases and control groups was statistically significant. ConclusionThe results of this study showed that injection of mice with a fraction of hydatid cyst fluid significantly inhibits the growth of mouse colon cancer and this inhibition may be related to effect of immune response to these antigens.

Taenia solium: Socioeconomic changes and prevalence evolution in four endemic communities in northwestern Mexico.

In Mexico, for the past 30 years, a continuous decrease in the incidence of clinical taeniosis/cysticercosis has been documented. This work aimed to determine the influence of improvement in socioeconomic conditions on the prevalence of Taenia solium in four endemic communities in northwestern Mexico. This study was carried out in two phases. First, documentary information (1989-2018) was collected about the prevalence of Theridion solium in the federal entity of Sinaloa State. Second, a pilot study was performed in four communities of Sinaloa, which had an endemic history of Taenia transmission. In each community, a risk factor questionnaire was applied, and serum and stool samples were collected for convenience in a non-probabilistic way. Anti-cysticercus antibodies and adult worm coproantigen were determined. The documentary analysis showed the incidence of taeniosis and cysticercosis to have decreased by 98 and 53%, respectively, while the human development index increased by 5% (1992-2017). Our data suggest that the risk of parasitic transmission is low, although female sex was a risk factor for reporting tremors or seizures (prevalence rate 2.1336, CI: 1.1821-3.8508) and background of tapeworm infection (prevalence rate 1.2893, CI: 0.9795-1.6972). No tapeworms or eggs were found while examining stool samples, but protozoa cysts were observed in four samples. Unexpectedly, only one of the 79 stool samples was positive for coproantigens. This positive result was confirmed in a second sample. However, the evaluation of a third sample was negative. No antibodies were found in human (n=377) or pig (n=69) samples. These data suggest parasite transmission has been interrupted and could be possibly associated with improving socioeconomic conditions. Further studies are needed to determine the real prevalence of zoonoses in Mexico.

Potent Inhibition of Macropinocytosis by Niclosamide in Cancer Cells: A Novel Mechanism for the Anticancer Efficacy for the Antihelminthic.

Niclosamide, a drug used to treat tapeworm infection, possesses anticancer effects by interfering with multiple signaling pathways. Niclosamide also causes intracellular acidification. We have recently discovered that the amino acid transporter SLC38A5, an amino acid-dependent Na+/H+ exchanger, activates macropinocytosis in cancer cells via amino acid-induced intracellular alkalinization. Therefore, we asked whether niclosamide will block basal and SLC38A5-mediated macropinocytosis via intracellular acidification. We monitored macropinocytosis in pancreatic and breast cancer cells using TMR-dextran and the function of SLC38A5 by measuring Li+-stimulated serine uptake. The peptide transporter activity was measured by the uptake of glycylsarcosine. Treatment of the cancer cells with niclosamide caused intracellular acidification. The drug blocked basal and serine-induced macropinocytosis with differential potency, with an EC50 of ~5 μM for the former and ~0.4 μM for the latter. The increased potency for amino acid-mediated macropinocytosis is due to direct inhibition of SLC38A5 by niclosamide in addition to the ability of the drug to cause intracellular acidification. The drug also inhibited the activity of the H+-coupled peptide transporter. We conclude that niclosamide induces nutrient starvation in cancer cells by blocking macropinocytosis, SLC38A5 and the peptide transporter. These studies uncover novel, hitherto unknown, mechanisms for the anticancer efficacy of this antihelminthic.

Tapeworm infection and secondary hemorrhagic shock found after cervical cancer surgery: a case report

Radical hysterectomy is an essential component of surgery for early-stage cervical cancer, and secondary hemorrhagic shock due to infection and vaginal stump necrosis after laparoscopic hysterectomy is very rare. Here we report a tapeworm infection and secondary hemorrhagic shock in a patient with cervical cancer after radical hysterectomy. On day two after cervical cancer surgery, this patient was found to have a long-term intestinal tapeworm infection and was discharged on postoperative day 11 after deworming on postoperative day 4. The patient entered the emergency operating room for hemorrhagic shock on day 15 after cervical cancer surgery; after hemodynamic resuscitation, gauze tamponade, and anti-infective treatment, the patient’s massive blood loss temporarily stopped. Two days later, the patient’s vaginal stump bled again repeatedly. Eventually, after 17 hours of surgery and hemodynamic resuscitation, the patient’s wound stopped bleeding profusely with ligation of the right internal iliac artery and branch arteries. This patient had a rare tapeworm infection that presented preoperatively but was detected only after cervical cancer surgery. We considered that preoperative anemia, long surgery time, surgical trauma, and intraoperative blood loss have significantly suppressed the patient’s immune response. A long-term intestinal tapeworm infection increased her susceptibility to bacteria, resulting in secondary pathogenic infection and postoperative blood loss. We advised that if a patient has combined parasitic infection and recurrent fever after a hysterectomy, it is necessary to prolong the hospital stay time, improve anemia, and increase immunity until the condition stabilizes before discharge.

Identification of Parasites on Fish Consumption at Several Fish Markets in Bogor City

This study aims to identify the types of parasites in fish consumed in several fish markets in Bogor City. This study used 100 consumption fish which included goldfish, tilapia, catfish, and mujahir fish. Sampling Fish were taken randomly at 5 fish markets in Bogor City, namely Gunung Batu Market, Pajajaran Market, Merdeka Market, Bogor Market, and Yasmin Market. Samples of fish organs examined were gills, fins and digestive organs (intestines). The examination results showed that the prevalence of consumption fish infected with parasites was 22% and included in the “Often” category. The highest prevalence of parasite infection was at Gunung Batu Market and Merdeka Market with a prevalence of 30% each. The types of parasites found were cestode worms namely Ligula intestinalis and Trematoda sub class Monogea namely Dactylogyrus sp and Gyrodactylus sp. Ligula intestinalis infects goldfish. Dactylogyrus sp and Gyrodactylus sp infected goldfish and mujair fish. Parasite infections in consuming fish at the Bogor City Fish Market are caused by dense fish density, poor water quality, optimal temperature for parasites and stress factors.

In vitro Scolicidal Effect of Ethonolic Extracts of Juglans regia and Carica papaya on Hydatid Cysts of Sheep and Goats from North Western Himalayas

Over a century ago, there are anecdotal stories of the use of therapeutic antihelminthic plants, such as papaya (Carica papaya) against cestode infections. Various studies have explored the efficacy of traditional plants in the inactivation of protoscolices and have reported the scolicidal effect of plants. Further, these plants have relatively lower side effects compared to chemotherapeutic agents and are suggested to be used for treatment of this disease in humans as well. Thus, in the present study the in vitro scolicidal effect of C. papaya and J. regia on Echinococcus granulosus protoscolices was explored from sheep, goat and human cysts in Jammu region of North Western Himalayas. The ethanolic extracts of J. regia and C. papaya showed significant scolicidal activity against E. granulosus, under in-vitro conditions with reference to the known standard drug “Praziquantel”. Against J. regia, highest mortality was observed at 30 mg/ml concentration at different exposure time as 10 min. (88.58%), 20 min. (91.24%), 30 min. (93.16%) and 40 min. (96.64%). Against C. papaya, highest mortality was observed at 30 mg/ml concentration at different exposure time as 10 min. (82.95%), 20 min. (85.83%), 30 min. (90.23%) and 40 min. (92.95%).

Prevalence and Identification of Cestode Species Recovered from Infected Organs of Lizardfish (Saurida undosquamis)

192 specimens of the Saurida undosquamis were collected from the local market of Misurata city in Libya. the prevalence rate of cestode helminths in examined fish Saurida undosquamis, was high. Based on the morphological and morphometric characterization represented two species of cestodes were isolated. Of the total examined fish (169, 88%) were infected with (Tetraphyllidean sp. and Trypanorhyncha sp. larvae). The density of cestode helminths was around 6810, and they were collected from the stomach, anterior and posterior parts of the intestine in all seasons, the liver and muscles showed no cestode infection, and a non-significant difference (P> 0.05) were found between cestode density infection of infected organs, this study found a high infection rate with Tetraphyllidean sp. and Trypanorhyncha sp. larvae in the Saurida undosquamis.

Intestinal cestode infection of Raillietina species in a 9 weeks old broiler in Umuahia, Abia State, Nigeria – A Case Report

This case report presents an intestinal cestode infection of Raillietina species in broilers. Live broiler birds that were 9 weeks old were seen to be infected with cestodes during a farm visit. Post mortem examination was carried out and samples of the intestines, live tapeworms and faeces were collected. On post mortem examination, the intestines were filled with whitish worms, the tapeworms were identified using microscopic examination as Raillietina species but no eggs of Raillietina species were observed following fecal examination. Although cestode infections with this species usually do not adversely affect humans following consumption, they are of economic importance as infections in birds can cause weight loss, growth problems and weakness. The essence of this report was to highlight and identify that cestode infections with Raillietina species can occur in broilers which is uncommon and the reason further investigation with microscopic examination was carried out following the negative result from the faecal examination

Natural variation in the binding pocket of a parasitic flatworm TRPM channel resolves the basis for praziquantel sensitivity

The drug praziquantel (PZQ) is the key clinical therapy for treating schistosomiasis and other infections caused by parasitic flatworms. A schistosome target for PZQ was recently identified- a transient receptor potential ion channel in the melastatin subfamily (TRPMPZQ)-however, little is known about the properties of TRPMPZQ in other parasitic flatworms. Here, TRPMPZQ orthologs were scrutinized from all currently available parasitic flatworm genomes. TRPMPZQ is present in all parasitic flatworms, and the consensus PZQ binding site was well conserved. Functional profiling of trematode, cestode, and a free-living flatworm TRPMPZQ ortholog revealed differing sensitives (~300-fold) of these TRPMPZQ channels toward PZQ, which matched the varied sensitivities of these different flatworms to PZQ. Three loci of variation were defined across the parasitic flatworm TRPMPZQ pocketome with the identity of an acidic residue in the TRP domain acting as a gatekeeper residue impacting PZQ residency within the TRPMPZQ ligand binding pocket. In trematodes and cyclophyllidean cestodes, which display high sensitivity to PZQ, this TRP domain residue is an aspartic acid which is permissive for potent activation by PZQ. However, the presence of a glutamic acid residue found in other parasitic and free-living flatworm TRPMPZQ was associated with lower sensitivity to PZQ. The definition of these different binding pocket architectures explains why PZQ shows high therapeutic effectiveness against specific fluke and tapeworm infections and will help the development of better tailored therapies toward other parasitic infections of humans, livestock, and fish.

Immunopathological Assessment of Hydatid Cyst and Cysticercus Tenuicollis Sonicated Protoscoilces Antigens in Mice

The present study was designed to investigate the cross-protection (protective immunity) between Hydatid cyst and Bladder worm and evaluate the immunologic response of both humerol and cellular immunity in mice. To achieve these goals, 120 mice were used and equally divided into four groups immunized subcutaneously with 2 doses of antigen at the first and 14 days of the experiment. Mice in the first group (n=30) were immunized with 0.3 ml of hydatid cyst sonicated protoscolex antigen. Those in the second group (n=30) were immunized s/c with 0.3 ml of Cysticercus tenuicollis sonicated protscolex antigen. The third group (n=30) was immunized with 0.3 ml of both antigens (0.15 + 0.15), and the fourth group was a control group in which the mice were intraperitoneally injected with 0.2 ml of phosphate buffer solution. At the end of the experiment (30 days), blood samples were taken from the hearts of mice in all groups after being anesthetized by intramuscular injection of Ketamine 60 mg/kg, and Xylazine 12 mg/kg for the assessment of mouse Interleukin-12, IgG, and tumor necrosis alpha levels. The skin test results 24 hours (day 28) post-immunization showed an increase in the skin thickness against both antigens in the treatments, compared to the control. However, there was a decrease at 48 hours (day 29) post-examination in all groups. The results of TNFα titer showed higher titer in the third group, compared to the first, second, and fourth groups. Interleukin 12 concentration showed a higher titer in the third group than in the first, second, and fourth groups. The IgG concentration showed higher titer in the third group compared to the first, second, and fourth groups. In conclusion, immunopathological studies have shown that Ags used in the study, induce humoral and cellular immunity, compared to each Ag alone, and the mixed antigens were much more immunogenic. This cross-reactivity and synergistic interactions between the two parasites may be the cause of their antigenic activities.

Comparative Analysis of the Distribution of the Cestode Infection Pathogen in the Internal Organs of Sheep in the Apsheron and Khizi Districts

During the research, cysts of Echinococcus granulosus and Cysticercus ovis were collected from the livers of sheep kept in Apsheron district. Echinococcus cysts were found in the whole liver and lungs of 302 out of 771 sheep examined. Among the study sites, Echinococcosiswas found in the villages of Tudar (47.9%), Jeyranbatan (47.6%), Altyagach (46.5%), Fatmayi (45.8%), and Kizilgazma (41.9%). Echinococcal cysts were collected from lungs and livers, and their distribution through organs was analyzed. As a result, in the livers, the intensity of invasion was 1-18 cysts, and in the lungs, it was 1-15 cysts. The high extent of invasion was recorded in the low mountain (45.1%) and foothills (42.5%) zones, and slightly less in the plain zone (32.7%). The infection rate of sheep increases from 28.3% in spring to 42.6% in summer and 48.5% in autumn, and these figures decrease again to 28.7% in winter. High infection with C. ovis species was recorded in Altyagach at 34.9%, Kizilgazma at 33.9%, and Tudar at 33.3%. In the villages of Mehdiabad, Sulutepe and Fatmayi, infection was not detected. C. ovis type is relatively low in the lowland zone with 8.8% EI, 1-8 II; in the areas belonging to the foothill zone, EI is 11.7%, EI is 1-8 units and is slightly higher; finally in the low mountainous zone, EI was 34.0%, EI was 1-19 units and was much higher. It was concluded from the research that the intensity of invasion due to both helminths was higher in Khizi district than Apsheron peninsula. Also, considering the altitude zones in the region, high intensity was recorded in the low mountainous zone.

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.

Hymenolepis diminuta Reduce Lactic Acid Bacterial Load and Induce Dysbiosis in the Early Infection of the Probiotic Colonization of Swiss Albino Rat.

Tapeworm infection continues to be an important cause of morbidity worldwide. Recent metagenomics studies have established a link between gut microbiota and parasite infection. The identification of gut probiotics is of foremost importance to explore its relationship and function with the parasite in the host. In this study, the gut content of hosts infected with tapeworm Hymenolepis diminuta and non-infected host gut were disected out to determine their Lactic acid bacterial (LAB) population in MRS agar and microbial community was analysed by metagenomics. The bacterial count was calculated on a bacterial counting chamber and their morphology was determined microscopically and biochemically. Further, to determine the safety profile antibiotic resistance test, antimicrobial, hemolytic activity, and adhesion capability were calculated. We found six dominant probiotic strains and a decrease in LAB load from 1.7-2.3 × 107 CFU/mL in the uninfected group to a range of 8.4 × 105 CFU/mL to 3.2 × 105 CFU/mL in the infected groups with respect to an increase in the parasite number from 10-18. In addition, we found a depletion in the probiotic relative abundance of Lactobacillus and an enrichment in potentially pathogenic Proteobacteria, Fusobacteria, and Streptococcus. Phylogenetic analysis of the six probiotics revealed a close similarity with different strains of L. brevis, L. johnsonii, L. taiwansis, L. reuteri, L. plantarum, and L. pentosus. Thus, this study suggests that the parasite inhibits probiotic colonization in the gut during its early establishment of infection inside the host.

The magic bullet: Niclosamide.

The term 'magic bullet' is a scientific concept proposed by the German Nobel laureate Paul Ehrlich in 1907, describing a medicine that could specifically and efficiently target a disease without harming the body. Oncologists have been looking for a magic bullet for cancer therapy ever since. However, the current therapies for cancers-including chemotherapy, radiation therapy, hormone therapy, and targeted therapy-pose either pan-cytotoxicity or only single-target efficacy, precluding their ability to function as a magic bullet. Intriguingly, niclosamide, an FDA-approved drug for treating tapeworm infections with an excellent safety profile, displays broad anti-cancer activity in a variety of contexts. In particular, niclosamide inhibits multiple oncogenic pathways such as Wnt/β-catenin, Ras, Stat3, Notch, E2F-Myc, NF-κB, and mTOR and activates tumor suppressor signaling pathways such as p53, PP2A, and AMPK. Moreover, niclosamide potentially improves immunotherapy by modulating pathways such as PD-1/PDL-1. We recently discovered that niclosamide ethanolamine (NEN) reprograms cellular metabolism through its uncoupler function, consequently remodeling the cellular epigenetic landscape to promote differentiation. Inspired by the promising results from the pre-clinical studies, several clinical trials are ongoing to assess the therapeutic effect of niclosamide in cancer patients. This current review summarizes the functions, mechanism of action, and potential applications of niclosamide in cancer therapy as a magic bullet.

Parasites and prey of the nursehound shark Scyliorhinus stellaris (Linnaeus, 1758): Insights into hidden trophic web interactions in the Mediterranean Sea.

The metazoan parasite community and the stomach contents of the nursehound shark Scyliorhinus stellaris from the Gulf of Naples (central Mediterranean Sea) were studied. The nursehound shark harboured a poor parasite community composed of a species of gill monogenean (Hexabothrium appendiculatum) and three intestinal cestode taxa (Acanthobothrium coronatum and two unidentified species of the genera Yamaguticestus and Scyphophyllidium), all represented by adult stages. Hosts were mostly parasitized by individuals of A. coronatum, which was the most abundant species and contributed to almost 80% of the total number of parasites found. Conversely, other trophically transmitted parasites (i.e., Yamaguticestus sp. and Scyphophyllidium sp.) showed low prevalence and abundance. The parasite infracommunity was poor, showing low values of species richness, total mean abundance, and diversity indices. Overall, 52 prey items belonging to 13 taxa were identified in the stomach contents. Cephalopods were the most important prey items (represented by nine taxa) and the most diverse and abundant group. In the multivariate space provided by a principal component of mixed data (PCAmix), nursehound sharks distributed along two main axes, related to individual traits (first axis) and stomach contents (including empty ones, second axis). A logistic regression based on the first two axes of the PCAmix showed a significant influence of host individual traits and, to a lesser extent, of stomach contents, regarding the probability of being infected by A. coronatum. Alongside specific traits already associated with parasites transmission, our results highlight the importance of cephalopods in transferring cestode infections through trophic web interactions in the top-predator nursehound shark.

Evaluating noninvasive methods for estimating cestode prevalence in a wild carnivore population.

Helminth infections are cryptic and can be difficult to study in wildlife species. Helminth research in wildlife hosts has historically required invasive animal handling and necropsy, while results from noninvasive parasite research, like scat analysis, may not be possible at the helminth species or individual host levels. To increase the utility of noninvasive sampling, individual hosts can be identified by applying molecular methods. This allows for longitudinal sampling of known hosts and can be paired with individual-level covariates. Here we evaluate a combination of methods and existing long-term monitoring data to identify patterns of cestode infections in gray wolves in Yellowstone National Park. Our goals were: (1) Identify the species and apparent prevalence of cestodes infecting Yellowstone wolves; (2) Assess the relationships between wolf biological and social characteristics and cestode infections; (3) Examine how wolf samples were affected by environmental conditions with respect to the success of individual genotyping. We collected over 200 wolf scats from 2018-2020 and conducted laboratory analyses including individual wolf genotyping, sex identification, cestode identification, and fecal glucocorticoid measurements. Wolf genotyping success rate was 45%, which was higher in the winter but decreased with higher precipitation and as more time elapsed between scat deposit and collection. One cestode species was detected in 28% of all fecal samples, and 38% of known individuals. The most common infection was Echinococcus granulosus sensu lato (primarily E. canadensis). Adult wolves had 4x greater odds of having a cestode infection than pups, as well as wolves sampled in the winter. Our methods provide an alternative approach to estimate cestode prevalence and to linking parasites to known individuals in a wild host system, but may be most useful when employed in existing study systems and when field collections are designed to minimize the time between fecal deposition and collection.

Niclosamide Inhibits Proliferation of Leukemia Cells and Synergizes with Chemotherapy

There have been significant advances in the treatment of acute childhood leukemia, which accounts for 25% of all childhood cancer cases. The 5-year survival rate for children with acute lymphoblastic leukemia (ALL) is 90%; however, there are few therapies available for relapsed disease. Pediatric acute myeloid leukemia (AML) has a 5-year survival rate of 65%, but current treatments are associated with significant morbidity and mortality. Less toxic and more effective therapies are needed to improve outcomes for patients with acute childhood leukemia. We previously showed that the cAMP-responsive element binding protein (CREB) plays an important role in leukemia cell proliferation and is overexpressed in blast cells from both ALL and AML patients. Niclosamide, an FDA-approved oral drug that has been used for decades to safely treat tapeworm infections, suppresses AML cell proliferation through inhibition of CREB activation and oxidative phosphorylation (Chae, et al. 2016). As a result of our preclinical studies, a Phase I clinical trial with niclosamide and cytarabine for relapsed/refractory pediatric AML is in progress. The effects of niclosamide in ALL cells or mechanisms of resistance to niclosamide have not been previously studied. The purpose of this study was to determine the antileukemic effects of niclosamide in ALL and to investigate additional synergistic effects of niclosamide in combination with drugs that are known to be effective against ALL or AML. To determine the effects of niclosamide on ALL proliferation, we treated two human B-ALL cell lines, Nalm6 and REH, and two human T-ALL cell lines, Jurkat and Loucy, with niclosamide alone or in combination with vincristine. Cells (2x104) were treated with 0.01-10µM niclosamide or 0.1% DMSO control. After 72 hours, CellTiter-Glo assays were performed to assess the concentration of drug that inhibited cell viability by 50% (IC50). The IC50 values for Nalm6 and REH were 0.45±0.13µM and 0.51±0.11µM, respectively (n=3). IC50 values for Jurkat and Loucy were 0.21±0.30µM and 0.58±0.11µM, respectively (n=3). Next, we investigated the potential synergistic effects between niclosamide and the chemotherapy drug, vincristine. The combination index (CI) values were calculated using the CompuSyn software (Chou-Talalay method). Simultaneous treatment of Nalm6 cells (ED75, 0.547; ED90, 0.316; ED95, 0.219; n=3) and Jurkat cells (ED75, 0.790; ED90, 0.686; ED95, 0.625; n=3) with niclosamide and vincristine had synergistic effects on cellular viability, indicating activity against both B-ALL and T-ALL. To study potential mechanisms of resistance to niclosamide, we performed a CRISPR/Cas9 library screen with HL60 human AML cells. Downregulation of BCL-2, an anti-apoptotic gene, sensitized cells to niclosamide treatment. Therefore, we investigated the effects of combining niclosamide with the BCL-2 inhibitor venetoclax in human AML cell lines, HL60 and U937, and in primary AML cells. CI values indicated synergy between niclosamide and venetoclax in HL60 cells (ED75, 0.251; ED90, 0.103; ED95, 0.063; n=5), U937 cells (ED75, 0.628; ED90, 0.614; ED95, 0.614; n=3), and patient sample 186 (ED75, 0.226; ED90, 0.301; ED95, 0.383; n=3). To investigate the effects of pretreatment with niclosamide, HL60 cells were cultured with niclosamide or vehicle for 3 days, followed by venetoclax for 3 days. Pretreatment with 300nM niclosamide significantly decreased the IC50 of venetoclax from 17.7±4.5nM to 1.87±0.46nM (n=3, p<0.05), indicating that niclosamide pretreatment sensitized HL60 cells to venetoclax. Our study established that both B-ALL and T-ALL cells are sensitive to niclosamide. Furthermore, niclosamide has synergistic effects when combined with standard chemotherapeutic agents such as vincristine and venetoclax. Our results demonstrate that niclosamide is a promising therapeutic agent for both AML and ALL and should be considered in future clinical trials.

Cestode Infection in the Digestive Tract of Ducks from Chicken Slaughterhouses in Bojong Gede and Cibinong Market, Bogor, West Java

Helminthiasis is a crucial gastrointestinal endoparasitic in ducks. This study aimed to determine the type and degree of infection of adult worms in the digestive tract of ducks at a chicken slaughterhouse in Bogor Regency, West Java. A total of 10 samples of the digestive tract of ducks were collected in this study. Five ducks were collected from a chicken slaughterhouse in Bojong Gede District and five other ducks were collected from a market in Cibinong City. Each intestinal sample was divided into 4 parts, i.e. the duodenum, jejunum, ileum and cecum. Adult cestode worms were stained with Semichon's Acetocarmine for identification. In result, three ducks were positive for endoparasite infection. The cestodes found were Raillietina cesticillus with 20% degree of infection in the jenjunum and ileum respectively. Hymenolepis nana with 20% degree of infection in the jejunum and ileum respectively. Choanotaenia infundibulum with 10% degree of infection in the ileum.