Herbal-based compounds: A review on treatments of cryptosporidiosis.

Cryptosporidium, a monoxenous apicomplexan coccidia, is a prevalent diarrhetic and an opportunistic agent, mainly in immunocompromised individuals. As there are few chemotherapeutic compounds that have limited efficacy, we need to identify new compounds or specific parasite targets for designing more potent drugs to treat cryptosporidiosis. Herbal products with low toxicity, environmental compatibility, wide therapeutic potential, and abundant resources can be considered alternatives for treatment. The current review tried to summarize the studies on plants or herbal bioactive constituents with anti-cryptosporidial activities. Based on constituents, plants act via different mechanisms, and further investigations are needed to clarify the exact mechanisms by which they act on the developmental stages of the parasite or host-parasite relationships.

Newly fabricated zinc oxide nanoparticles loaded materials for therapeutic nano delivery in experimental cryptosporidiosis.

Cryptosporidiosis is a global health problem that threatens the lives of immunocompromised patients. This study targets to fabricate and investigate the efficiency of zinc oxide nanoparticles (ZnO-NPs), nitazoxanide (NTZ)-loaded ZnO-NPs, and Allium sativum (A. sativum)-loaded ZnO-NPs in treating cryptosporidiosis. Further FTIR, SEM, XRD, and zeta analysis were used for the characterization of ZnO-NPs and loaded materials. The morphology of loaded materials for ZnO-NPs changed into wrapped layers and well-distributed homogenous particles, which had a direct effect on the oocyst wall. The charge surface of all particles had a negative sign, which indicated well distribution into the parasite matrix. For anti-cryptosporidiosis efficiency, thirty immunosuppressed Cryptosporidium parvum-infected mice, classified into six groups, were sacrificed on the 21st day after infection with an evaluation of parasitological, histopathological, and oxidative markers. It was detected that the highest reduction percent of Cryptosporidium oocyst shedding was (81.5%) in NTZ, followed by (71.1%) in A. sativum-loaded ZnO-NPs-treated groups. Also, treatment with A. sativum and NTZ-loaded ZnO-NPs revealed remarkable amelioration of the intestinal, hepatic, and pulmonary histopathological lesions. Furthermore, they significantly produced an increase in GSH values and improved the changes in NO and MDA levels. In conclusion, this study is the first to report ZnO-NPs as an effective therapy for treating cryptosporidiosis, especially when combined with other treatments that enhance their antioxidant activity. It provides an economical and environment-friendly approach to novel delivery synthesis for antiparasitic applications.

Study on the Therapeutic Effect of Coconut Oil Extracts as An Alternative Medicinal Plant in Cryptosporidium Infected Mice.

OBJECTIVE: Cryptosporidiosis caused by Cryptosporidium sp. is a globally spreading disease. Nowadays, new researches are moving towards an effective treatment without side effects, especially for young and immune-compromised patients. The current study was designed to evaluate the therapeutic effect of the coconut oil extracts as an alternative medicinal plant in Cryptosporidium infected immunocompromised mice. METHODS: Sixty white albino mice were classified into six groups; Group I: Infected with Cryptosporidium oocysts treated with Nitazoxanide, Group II: Infected with Cryptosporidium oocysts and treated with coconut water extract, Group III: Infected with Cryptosporidium oocysts and treated with coconut Hexan extract, Group IV: Infected with Cryptosporidium oocysts and treated with coconut ethanol extract, Group V: Positive control, Group VI: Negative control. Stool samples were collected and examined; histopathological and immune-histochemical assessment using anti caspase-3 and anti CDX2 monoclonal antibodies were performed. RESULTS: Coconut oil extracts results revealed a significant decrease of oocyst count, correlated with an amelioration of histopathological and confirmed by immunohistochemical changes in ileal tissue. CONCLUSION: The present study has opened fresh avenues for development of natural therapy like coconut oil extracts, which have a potential therapeutic efficacy against Cryptosporidiosis. That was confirmed by different methodologies, parasitological examination, histopathological examination, and immunohistochemical assays. It paves the way for being a promising anti-parasitic agent for infection eradication. However, further studies are still required to gain more knowledge about different coconut extracts in order to reach the best treatment efficacy.

The controverted therapeutic efficacy of Allium sativum and Artemisia herba-alba extracts on Cryptosporidium-infected mice.

INTRODUCTION: Cryptosporidiosis has become an issue of great interest being life-threatening among immunocompromised hosts worldwide. This study explored the curative effect of Allium sativum (garlic) and Artemisia herba-alba ethanolic extract versus Nitazoxanide drug on both immunocompetent and immunosuppressed-Cryptosporidium experimentally-infected mice. METHODOLOGY: One hundred male Swiss albino mice were divided into the following groups: (GI) non-infected non-treated group, (GII) infected non-treated group, (GIII) garlic treated group, (GIV) A. herba-alba treated group, (GV) Nitazoxanide treated group, each group subdivided into two subgroups (a) Immunocompetent, (b) immunosuppressed. The assessment was performed by parasitological counting of fecal oocysts, histological examination of intestinal tissue, immunological detection of interferon-gamma levels in mice sera, and ultrastructural study by transmission electron microscopy. RESULTS: Garlic and A. herbal-alba extracts showed a decrease in the mean oocyst counts through all days of follow-up. This was associated with significant up-regulation of interferon-gamma cytokine levels in serum and histological improvement in intestinal tissues of mice compared to control groups and the results were confirmed by transmission electron microscopy. The highest efficacy was obtained by garlic, then by A. herbal-alba extracts followed by Nitazoxanide treated group; where the immunocompetent groups showed better improvement than immunosuppressed ones. CONCLUSIONS: Garlic has a perfect effect as a promising therapeutic agent against Cryptosporidiosis and therefore validates their traditional use in parasitic infections. Accordingly, it may offer a good option for cryptosporidium treatment in immunocompromised patients. They could be used as a natural safe product for the preparation of a new therapeutic agent.

Investigation of the prophylactic and therapeutic effectiveness of oral thyme extract in rats experimentally infected with cryptosporidium parvum.

In this study, the prophylactic and therapeutic activities of thyme extract at different concentrations against experimental Cryptosporidium parvum infection in immunosuppressed rats were investigated. Thyme extract was prepared at four different concentrations (10%, 30%, 50%, and 100%) and administered as a single oral dose of 1 mL for evaluation of its prophylactic efficacy. Five consecutive days after infection was detected in all rats, therapeutic evaluations were also performed. According to the results obtained by daily counting of oocysts in stools, the prophylactic and therapeutic effects of thyme extract administration were significant in comparison to the control group (P˂0.01). Oocyst shedding continued in the control group at high numbers from the beginning to the end of the study, while oocyst counts in the prophylaxis groups remained low throughout the study. On the other hand, oocyst excretion rates were high in the therapeutic groups and decreased rapidly after thyme extract administration. At the end of the study, oocyst excretion had completely stopped for some rats administered thyme extract. There was no group in which oocyst shedding ceased for all rats. No significant differences were observed in the therapeutic or prophylaxis groups regarding the doses administered (P > 0.01). Renal and hepatic functions were monitored by measuring urea, creatinine, alanine transaminase, and aspartate transaminase levels ​​before and after thyme extract administration. As a result, it was concluded that oral thyme extract administration at the doses applied in this study is effective and safe in the prophylactic and therapeutic treatment of experimental cryptosporidiosis in rats.

Therapeutic Potency of Ginger, Garlic, and Pomegranate Extracts Against Cryptosporidium parvum-Mediated Gastro-Splenic Damage in Mice.

PURPOSE: Cryptosporidium parvum is a protozoan parasite infecting most mammalian hosts and causing major health issues. The present study investigated the efficacy of ginger (Zingiber officinale), garlic (Allium sativum), and pomegranate (Punica granatum) peel extracts on the development and progression of experimental cryptosporidiosis in mice. METHODS: Eighty-two mice were assigned to 6 groups: control, infected non-treated, metronidazole (MTZ), ginger, garlic, and pomegranate. The control group topically received no treatments. The infected non-treated group was experimentally infected by 104 C. parvum oocysts per mouse using a stomach tube. The MTZ group was infected with C. parvum oocysts combined with MTZ (50 mg/kg b.w./day). The ginger, garlic, and pomegranate groups daily received different plant extracts at doses of 100 mg/kg BW, 50 mg/kg BW, and 3 gm/kg BW, respectively, followed by infection with C. parvum oocysts. All treatments were applied orally one day after the infection for continuous 30 days. RESULTS: Histopathological and immunohistochemical examinations for P53 and caspase-3 expressions in stomach and spleen tissues showed that MTZ and garlic-treated mice had a more significant effect on infected mice. CONCLUSION: The garlic extract was found to exert a more pronounced effect on infected mice compared with the other treatments as well as to improve health. Garlic extracts, therefore, represent an effective and natural therapeutic alternative for the treatment of cryptosporidiosis with low side effects and without drug resistance.

Potential effect of methanol leaf extracts from three Podocarpus species on experimental cryptosporidiosis.

Cryptosporidiosis causes diarrhea in both immunocompetent and immunocompromised individuals, with acute manifestations occurring particularly in children and the elderly. Up till now, there is no curative therapy for cryptosporidiosis, so discovery of new classes of drugs are of great importance. This study aimed to examine the effect of methanol leaves extracts of the three Podocarpus species; P. macrophyllus (Thunb.), P. gracilior (Pilg.) and P. elongatus (Aiton) L' Hér. ex Pers and their combination on Cryptosporidium parvum (C. parvum) in experimentally infected mice in comparison with the commercially used drug, Nitazoxanide. As well as spectrophotometric estimation of the total phenolic and flavonoid content of these extracts was done. Results revealed that treatment with these three Podocarpus extracts and their combination showed a significant reduction of the number of C. parvum oocyst shed in the stool of infected mice compared to infected control group and Nitazoxanide- infected treated group at P < 0.001. The combination of the three Podocarpus extracts was the most effective treatment showing the lowest number of oocysts shedding in comparison with other used extracts and Nitazoxanide. Histopathological inspection of sections from ilium and colon displayed signs of improvement after treatment with P. macrophyllus and P. gracilior extracts and more remarkable improvement when the three extracts were combined. It was concluded that the three Podocarpus species extracts used in this study had a promising anti-Cryptosporidium activity especially when they were combined.

Evaluation of the efficacy of cow colostrum in the treatment and its effect on serum proteomes in calves with cryptosporidiosis.

This study aimed to investigate the effect of using high-quality colostrum in addition to paromomycin on the treatment outcomes and serum proteomes of calves naturally affected by cryptosporidiosis. Thirty Holstein calves infected with only Cryptosporidium spp. were divided into three equal groups. Calves in the PC group received paromomycin orally at a dose of 100 mg/kg once daily for 5 days. Calves in PCOL and PBCOL groups received 250 ml colostrum 3 h after feeding twice a day for 3 days. The PBCOL group was also given 6 g of sodium bicarbonate 15 min before colostrum administration. While the fecal scores of all calves were evaluated daily for 10 days from the initiation of the treatment, fecal oocyst counts were determined on the 0, 1, 2, 3, 5, 7, and 10th days. Brix%, total protein (TP), immunoglobulin G (IgG), gamma-glutamyltransferase (GGT) levels, and proteomic analyses were performed on the 0 and 3rd days' sera. Considering pretreatment values, fecal scores (8th, 2nd, and 2nd day), and fecal oocyst counts (10th, 3rd, and 2nd day) improved in a significantly (p < 0.05) shorter time in the colostrum groups than in the control group. By serum proteomic analysis, 99, 93, and 83 proteomes were detected in PC, PCOL, and PBCOL groups, respectively. Although the significant changes in any protein in Group PC were absent, significant changes were observed in Alpha-1B-glycoprotein (A1BG), Zinc transporterZIP11 (S39AB), Cathelicidin-1 (CTHL1), Actin_ cytoplasmic-1 (ACTB), and Apolipoprotein A-IV (APOA4) proteins in Group PCOL and Alpha-1-antiproteinase (A1AT), Serum amyloid A protein (SAA), Actin-cytoplasmic-2 (ACTG), Protein HP-20 homolog (HP20) proteins in Group PBCOL with colostral treatment, which indicated that the use of colostrum had an effect on calf serum proteomes. The more pronounced healing and shorter clinical improvement time in the colostrum groups especially colostrum with sodium bicarbonate revealed that these proteomes have positive effects in the treatment with their systemic and local effects in the intestines.

Garlic (Allium sativum Linnaeus) improved inflammation and reduced cryptosporidiosis burden in immunocompromised mice.

ETHNOPHARMACOLOGICAL RELEVANCE: For thousands of years, garlic (Allium sativum Linnaeus) has been consumed in food and health by numerous civilizations. Cryptosporidium (C.) parvum is an apicomplexan parasite that causes a gastrointestinal disease, with the most common symptoms being watery diarrhea. Although several substances have been tried for its anti-cryptosporidial action, there is no effective treatment for Cryptosporidium disease, especially in immunocompromised individuals. The present study aimed firstly to characterize the bio-active compounds in Allium sativum L. and secondly to evaluate its efficacy as a therapy for cryptosporidiosis especially in immunocompromised mice. MATERIALS AND METHODS: This was accomplished by evaluating the parasitological and histopathological parameters in the experimentally infected immunocompetent and immunocompromised mice. Also, the cytokine profile during the experimental time was recorded through the measuring of T helper (h)1, Th2 and Th17 cells cytokines. Immunosuppressed mice were given 0.25 µg/g per day of dexamethasone orally, before infection with Cryptosporidium parvum oocysts, for fourteen consecutive days. Starting 10 days post infection (PI), nitazoxanide (100 mg/kg per day) or Allium sativum (50 mg/kg per day) was given orally for fourteen consecutive days. RESULTS: Our results showed that oocyst shedding, on the 32nd day PI, in immunocompromised infected group treated with Allium sativum (354.11, 99.35% PR) showed a significant decrease when compared to its corresponding group treated with nitazoxanide (4369.14, 92.05% PR). On the 32nd day PI, all cytokines levels have been decreased to levels that were similar to those of their uninfected corresponding control groups; also, the histopathological changes and the loss in animals' body weight had been improved. Treatment with nitazoxanide did not result in infection clearance or a reduction in the increased cytokines' levels. CONCLUSION: Allium sativum L. displayed high efficacy as a potential therapeutic agent against Cryptosporidium, which supports its traditional usage in parasite diseases.

Antiparasitic and immunomodulating effects of nitazoxanide, ivermectin and selenium on Cryptosporidium infection in diabetic mice.

The present work aims to investigate the antiparasitic and the immunomodulating effects of nitazoxanide (NTZ) and ivermectin (IVC) alone or combined together or combined with selenium (Se), on Cryptosporidium infection in diabetic mice. The results revealed that the combined NTZ and IVC therapy achieved the highest reduction of fecal oocysts (92%), whereas single NTZ showed the lowest reduction (63%). Also, adding Se to either NTZ or IVC resulted in elevation of oocyst reduction from 63% to 71% and from 82% to 84% respectively. All treatment regimens, with the exception of NTZ monotherapy, showed a significant improvement in the intestinal histopathology, the highest score was in combined NTZ and IVC therapy. The unique results of immunohistochemistry in this study showed reversal of the normal CD4/CD8 T cell ratio in the infected untreated mice, however, following therapy it reverts back to a normal balanced ratio. The combined (NTZ+ IVC) treatment demonstrated the highest level of CD4 T cell expression. Taken together, NTZ and IVC combined therapy showed remarkable anti-parasitic and immunostimulatory effects, specifically towards the CD4 population that seem to be promising in controlling cryptosporidiosis in diabetic individuals. Further research is required to explore other effective treatment strategies for those comorbid patients.

Use of a natural herbal-based feed additive containing isoquinoline alkaloids in newborn calves with cryptosporidiosis.

Cryptosporidium infections are one of the most prevalent causes of diarrhea in calves and considered to be one of the major sources of economic loss in livestock production. A global trend is currently underway, in identifying natural and sustainable alternatives to support animal husbandry and production. Isoquinoline alkaloids are known for their anti-inflammatory, antimicrobial, and antioxidant properties in the promotion of gut health. Thus, an experiment was designed to evaluate the effects of natural, herbal-based feed isoquinoline alkaloids to support calves experimentally inoculated with Cryptosporidium parvum. Twenty-six calves were randomly divided into control (CN) (n = 13) and treatment (SG) (n = 13) groups. The SG group received 5 g of feed additive in every milk feeding from 1 to 21 days of age. The CN group received milk without any additives. All calves were orally inoculated on the third day of life with 1 × 106Cryptosporidium parvum oocysts. The animals were evaluated daily, from 3 to 30 days of age, for the occurrence, duration, and intensity of diarrhea. Calves with a base deficit of ≥ 9 mEq/L were hydrated to aid recovery. The SG calves showed a higher average weight gain between 14 and 21 days of age, without mortality and with reduced intensity and duration of diarrhea. In contrast, calves in the CN group showed more serious acid-base disorders, required more hydration support, and had a mortality rate of 15.4 %. These results showed that calves supplemented with isoquinoline alkaloids had decreased intensity and duration of symptoms, reduced requirement for supportive therapy, and prevented mortality among animals.

[Identification of potential targets and synergistic mechanism of Kushen Decoction for the treatment of cryptosporidiosis].

OBJECTIVE: To explore the potential targets and synergistic mechanisms of Kushen Decoction for the treatment of cryptosporidiosis using network pharmacology and molecular docking methods. METHODS: The main active ingredients of Kushen Decoction were captured from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TC-MSP) and the Universal Protein Resource (UniProt) database, and the potential targets were predicted. In addition, the active ingredients of Kushen Decoction that were not included in the TCMSP database were retrieved in CNKI, WanFang Data, CBM, PubMed and Web of Science databases, and the target genes of all supplemented active ingredients were predicted using the online TargetNet database. Network construction and analysis were performed using the Cytoscape software, and cryptosporidiosis-related targets were retrieved in the Comparative Toxicogenomics Database and GeneCards database. The protein-protein interaction (PPI) network was created using the STRING database, and the DAVID database was used for GO enrichment and KEGG pathway analyses. The tissue distribution of key targets was investigated using the BioGPS database, and the AutoDockTools software was employed to verify the molecular docking results. RESULTS: A total of 38 active ingredients of Kushen Decoction were screened, and the core ingredients included quercetin, (+)-14α-hydroxymatrine and apigenin. A total of 831 targets of Kushen Decoction and 512 cryptosporidiosis-related targets were predicted, and PPI network analysis revealed 69 key targets, including AKT1, TNF and IL-6. There were 303 biological processes, 46 molecular functions and 29 cellular components involved in the treatment of cryptosporidiosis with Kushen Decoction, and 13 KEGG pathways played a therapeutic role in the synergistic mechanisms of multiple targets, such as Toll-like receptor (TLR), nuclear factor kappa B(NF)-κB, nucleotide binding oligomerization domain like receptor (NLR) signal pathways. The core targets were mainly distributed in the hematologic and immune systems. Molecular docking analysis showed that the binding energy between active ingredients and key targets were all less than 0 kJ/mol, indicating the strong binding of ligands to receptors. CONCLUSIONS: The active ingredients of Kushen Decoction, such as quercetin, (+)-14α-hydroxymatrine and apigenin, may act on targets like AKT1, TNF, IL-6 to modulate TLR, NLR and NF-κB signaling pathways to play a synergistic role in the treatment of cryptosporidiosis in the hematologic and immune system.

In vitro and in vivo anti-Cryptosporidium and anti-inflammatory effects of Aloe vera gel in dexamethasone immunosuppressed mice.

Cryptosporidiosis has been considered as a serious diarrheal disease, especially in immunodeficient patients, where they failed to clear the infection leading to several consequences of infection (i.e death). The role of cell mediated immunity in clearing the infection was demonstrated by the increased susceptibility of HIV/AIDS patients to infection. To date, no specific treatment has been proven for cryptosporidiosis in immunodeficient patients. The study aimed to evaluate the efficacy of Aloe vera gel for the treatment of cryptosporidiosis in immunocompetent and dexamethasone immunosuppressed mice in comparison to that of nitazoxanide. Mice were orally administrated with Aloe vera gel, in a daily dose of 250 mg/L in drinking water, for 14 consecutive days post infection. Parasitological, molecular and immunological measurements were recorded on the 7th, 14th, 21st and 32nd days post infection. Our in vitro results showed that 250 mg/L of prepared gel achieved the highest parasitic reduction. The body weights of Aloe vera treated mice on the 21st and 32nd day post infection, either in immunocompetent or immunosuppressed groups, were nearly the same as those of their corresponding control groups. Aloe vera gel succeeded in clearing cryptosporidiosis with a percent reduction of 100% in immunocompetent mice and 99.67% in immunosuppressed mice. The anti-inflammatory effect of Aloe vera reduced the levels of IFN-γ, IL-4, -6 and -17. The success of Aloe vera gel, in clearing cryptosporidiosis in immunosuppressed mice, was obvious either from the reduction of Cryptosporidium DNA or the oocysts in stool samples; and from the improvement of histopathological sections.

Anticryptosporidium Efficacy of Olea europaea and Ficus carica Leaves Extract in Immunocompromised Mice Associated with Biochemical Characters and Antioxidative System.

Cryptosporidiosis is caused by an opportunistic protozoan parasite (Cryptosporidium parvum and C. hominis) known as a parasite of humans, especially children and immunocompromised patients. The current study was designed to evaluate the therapeutic efficacy of a mixture of fig and olive leaf extracts as an alternative medicinal plant. Parasitological examination for oocysts in the stool and histopathological alterations in the small intestines were examined. Additionally, biochemical analyses of liver and kidney functions in addition to antioxidant parameters such as superoxide dismutase (SOD), glutathione peroxidase (GSH) and catalase (CAT) in the plasma were evaluated. Our results showed that marked reduction in oocysts shedding and amelioration in intestinal histopathological changes and hepatic or renal functions were detected in all treated groups compared to the control infected group. Additionally, the treated groups with tested extracts at ratios 1:3 and 1:5 showed a significant decrease in the number of oocysts compared to the other treated groups. Results exhibited a significant increase in the plasma SOD, CAT and GSH levels in treated groups compared to the infected control one. This study suggested that a mixture of fig and olive leaf extracts is a convenient promising therapeutic agent for Cryptosporidiosis.

The Efficacy of Citrus maxima Peels Aqueous Extract Against Cryptosporidiosis in Immunecompromised Mice.

INTRODUCTION:  Cryptosporidiosis is an opportunistic globally distributed parasitic disease caused by protozoan Cryptosporidium where its development is closely related to the host's immune status. New therapeutic agents are a high priority as chemotherapeutics are impractical and vaccines are unavailable for young as well as immune-compromised patients or animals. The current study was designed to evaluate the therapeutic effect of the internal white (albedo) and external yellow (flavedo) peels of Citrus maxima (C. maxima) as an alternative medicinal plant. MATERIALS AND METHODS : Parasitological examination for oocysts in the stool was determined. Histopathological alterations and immunohistochemical expression of APC and cyclin D1 as well as an assessment of interferon-γ (IFN-γ) and interleukin 1ß (IL-1ß) in ileal tissues was carried out. In addition, the biochemical examination of serum albumin, globulin and liver enzymes were evaluated. Results revealed a significant decrease of oocysts count correlated with an amelioration of histopathological and immunohistochemical changes in ileal tissue with an enhancement of liver enzymes and inflammatory cytokines levels. CONCLUSION:  It could be concluded that treatment with C. maxima peel extracts have a potential therapeutic and an immunoregulatory efficacy against Cryptosporidiosis. Obtained results showed that the white peel was found to have more immunological effect that could significantly enhance inflammatory cytokines response towards normal status. Hence, it can be used in the daily animal diet to give protective effects against infection.

Antiparasitic and immunomodulating effects of nitazoxanide, ivermectin and selenium on Cryptosporidium infection in diabetic mice / Efeitos antiparasitários e imunomodulantes da nitazoxanida, ivermectina e selênio sobre a infecção por Cryptosporidium em camundongos diabéticos

Abstract The present work aims to investigate the antiparasitic and the immunomodulating effects of nitazoxanide (NTZ) and ivermectin (IVC) alone or combined together or combined with selenium (Se), on Cryptosporidium infection in diabetic mice. The results revealed that the combined NTZ and IVC therapy achieved the highest reduction of fecal oocysts (92%), whereas single NTZ showed the lowest reduction (63%). Also, adding Se to either NTZ or IVC resulted in elevation of oocyst reduction from 63% to 71% and from 82% to 84% respectively. All treatment regimens, with the exception of NTZ monotherapy, showed a significant improvement in the intestinal histopathology, the highest score was in combined NTZ and IVC therapy. The unique results of immunohistochemistry in this study showed reversal of the normal CD4/CD8 T cell ratio in the infected untreated mice, however, following therapy it reverts back to a normal balanced ratio. The combined (NTZ+ IVC) treatment demonstrated the highest level of CD4 T cell expression. Taken together, NTZ and IVC combined therapy showed remarkable anti-parasitic and immunostimulatory effects, specifically towards the CD4 population that seem to be promising in controlling cryptosporidiosis in diabetic individuals. Further research is required to explore other effective treatment strategies for those comorbid patients.

Resumo O presente trabalho tem como objetivo investigar os efeitos anti-parasitários e imunomodulantes da nitazoxanida (NTZ) e ivermectina (IVC), isoladas ou em associação, e do selênio (SE), associado à NTZ ou à IVC, sobre a infecção por Cryptosporidium em camundongos diabéticos. Os resultados revelaram que a terapia combinada com NTZ e IVC resultou em maior redução de oocistos fecais, enquanto a NTZ isolada mostrou a menor redução de oocistos fecais (63%). Além disso, a associação do SE com a NTZ ou IVC resultou em redução do número de oocistos fecais de 63% para 71% e de 82% para 84%, respectivamente. Todos os tratamentos, com exceção da monoterapia com NTZ, mostraram uma melhora significativa nos índices relacionados à histopatologia intestinal. Os resultados da imuno-histoquímica mostraram reversão da razão celular CD4/CD8 T normal nos camundongos infectados não tratados, no entanto, após a terapia, houve retorno à razão celular CD4/CD8 T normal. O tratamento combinado (NTZ+ IVC) demonstrou o mais alto nível de expressão celular CD4 T. Em conclusão, a terapia combinada com NTZ e IVC mostrou efeitos anti-parasitários e imunoestimuladores notáveis, especificamente para a população CD4, que parecem ser promissores para o controle da criptosporidiose em indivíduos diabéticos.

Identification of a potent benzoxaborole drug candidate for treating cryptosporidiosis.

Cryptosporidiosis is a leading cause of life-threatening diarrhea in young children and causes chronic diarrhea in AIDS patients, but the only approved treatment is ineffective in malnourished children and immunocompromised people. We here use a drug repositioning strategy and identify a promising anticryptosporidial drug candidate. Screening a library of benzoxaboroles comprised of analogs to four antiprotozoal chemical scaffolds under pre-clinical development for neglected tropical diseases for Cryptosporidium growth inhibitors identifies the 6-carboxamide benzoxaborole AN7973. AN7973 blocks intracellular parasite development, appears to be parasiticidal, and potently inhibits the two Cryptosporidium species most relevant to human health, C. parvum and C. hominis. It is efficacious in murine models of both acute and established infection, and in a neonatal dairy calf model of cryptosporidiosis. AN7973 also possesses favorable safety, stability, and PK parameters, and therefore, is an exciting drug candidate for treating cryptosporidiosis.

Curcumin: A promising treatment for Cryptosporidium parvum infection in immunosuppressed BALB/c mice.

Members of the genus Cryptosporidium are frequent protozoan pathogens in humans and a wide range of animals. There is no consistently effective treatment against cryptosporidiosis, especially in immunodeficient patients. The present study was carried out to study the therapeutic effects of curcumin against cryptosporidiosis in immunosuppressed BALB/c mice. Mice were divided into five groups and immunosuppressed by dexamethasone. Three groups were inoculated with C. parvum oocysts, administered with curcumin, paromomycin, and without treatment. The reminders were regarded as controls. The oocysts in the fecal smear were counted daily. At days 0, 3, 7, and 11 post-treatment, the mice were sacrificed, and the efficacy of drugs was evaluated by comparing the histopathological alterations in jejunum and ileum, measuring the total antioxidant capacity, and malondialdehyde in the affected tissues. The infection was completely eliminated in the curcumin-treated group, and oocyst shedding stopped with no recurrence after drug withdrawal. On the contrary, paromomycin was unable to eliminate C. parvum infection completely, and oocyst shedding continued even 10 days after the drug withdrawal. Based on these findings, curcumin can be a trustworthy compound for the elimination of infection in immunosuppressed hosts. Further evaluation to find its accurate mechanism of action should be considered.

The ReFRAME library as a comprehensive drug repurposing library and its application to the treatment of cryptosporidiosis.

The chemical diversity and known safety profiles of drugs previously tested in humans make them a valuable set of compounds to explore potential therapeutic utility in indications outside those originally targeted, especially neglected tropical diseases. This practice of "drug repurposing" has become commonplace in academic and other nonprofit drug-discovery efforts, with the appeal that significantly less time and resources are required to advance a candidate into the clinic. Here, we report a comprehensive open-access, drug repositioning screening set of 12,000 compounds (termed ReFRAME; Repurposing, Focused Rescue, and Accelerated Medchem) that was assembled by combining three widely used commercial drug competitive intelligence databases (Clarivate Integrity, GVK Excelra GoStar, and Citeline Pharmaprojects), together with extensive patent mining of small molecules that have been dosed in humans. To date, 12,000 compounds (∼80% of compounds identified from data mining) have been purchased or synthesized and subsequently plated for screening. To exemplify its utility, this collection was screened against Cryptosporidium spp., a major cause of childhood diarrhea in the developing world, and two active compounds previously tested in humans for other therapeutic indications were identified. Both compounds, VB-201 and a structurally related analog of ASP-7962, were subsequently shown to be efficacious in animal models of Cryptosporidium infection at clinically relevant doses, based on available human doses. In addition, an open-access data portal (https://reframedb.org) has been developed to share ReFRAME screen hits to encourage additional follow-up and maximize the impact of the ReFRAME screening collection.