Anthelmintic Activity of Yeast Particle-Encapsulated Terpenes

Mirza, Zeynep; Soto, Ernesto; Hu, Yan; Nguyen, Thanh-Thanh; Koch, David; Aroian, Raffi V.; Ostroff, Gary R.

doi:10.3390/molecules25132958

Cited by 21 publications

(27 citation statements)

References 42 publications

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“…Spore lysates (SLs; HD1 Cry deficient strains) and spore-crystal lysates (SCLs; HD1 strains transformed with Cry5B plasmid) were prepared using standard methods (40,45) For experiments in Table S1, Bt 407 ∆spo0A::kan cells transferred with pHY159 were grown in Luria broth overnight at 30°C in 10 µg/mL erythromycin to saturation. The next day, 100 µL of terpenes (49) were added to 100 µL of overnight culture to a final volume of 1 mL in Luria broth plus erythromycin. Tubes were shaken overnight at 30°C.…”

Section: Spore and Spore Crystal Lysatesmentioning

confidence: 99%

Recombinant Paraprobiotics as a New Paradigm for Treating Gastrointestinal Nematode Parasites of Humans

Abraham

Gazzola

et al. 2020

Preprint

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Gastrointestinal nematodes (GINs) of humans, e.g., hookworms, negatively impact childhood growth, cognition, nutrition, educational attainment, income, productivity, and pregnancy. Hundreds of millions of people are targeted with mass drug administration (MDA) of donated benzimidazole (BZ) anthelmintics. However, BZ efficacy against GINs is suboptimal, and reduced/low efficacy has been seen. Developing an anthelmintic for human MDA is daunting: it must be safe, effective, inexpensive, stable without a cold chain, and massively scalable. Bacillus thuringiensis (Bt) crystal protein 5B (Cry5B) has anthelmintic properties that could fill this void. Here we develop an API (Active Pharmaceutical Ingredient) form of Bt Cry5B compatible with MDA. We expressed Cry5B in asporogenous Bt during vegetative phase, forming cytosolic crystals. These Bacteria with Cytosolic Crystals (BaCC) were rendered inviable (inactivated BaCC or IBaCC) with food-grade essential oils. IBaCC potency was validated in vitro against nematodes. IBaCC was also potent in vivo against human hookworm infections in hamsters. IBaCC production was successfully scaled to 350 liters at a contract manufacturing facility. A simple fit-for-purpose formulation to protect against stomach digestion and powdered IBaCC were successfully made and used against GINS in hamsters and mice. A pilot histopathology study and blood chemistry workup showed that five daily consecutive doses of 200 mg/kg Cry5B IBaCC (the curative single dose is 40 mg/kg) was non-toxic and completely safe. IBaCC is a safe, inexpensive, highly effective, easy-to-manufacture, and scalable anthelmintic that is practical for MDA and represents a new paradigm for treating human GINs.

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Section: Spore and Spore Crystal Lysatesmentioning

confidence: 99%

Recombinant Paraprobiotics as a New Paradigm for Treating Gastrointestinal Nematode Parasites of Humans

Abraham

Gazzola

et al. 2020

Preprint

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“…It is not clear which BGCs could be responsible for this activity. A possibility could be represented by the terpene clusters, as it is long known that a few terpenes isolated from plants have shown anthelmintic activities [ 78 ]. Further studies are required to identify the responsible molecules.…”

Section: Resultsmentioning

confidence: 99%

Isolation and Characterization of Strain Exiguobacterium sp. KRL4, a Producer of Bioactive Secondary Metabolites from a Tibetan Glacier

et al. 2021

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Extremophilic microorganisms represent a unique source of novel natural products. Among them, cold adapted bacteria and particularly alpine microorganisms are still underexplored. Here, we describe the isolation and characterization of a novel Gram-positive, aerobic rod-shaped alpine bacterium (KRL4), isolated from sediments from the Karuola glacier in Tibet, China. Complete phenotypic analysis was performed revealing the great adaptability of the strain to a wide range of temperatures (5–40 °C), pHs (5.5–8.5), and salinities (0–15% w/v NaCl). Genome sequencing identified KRL4 as a member of the placeholder genus Exiguobacterium_A and annotation revealed that only half of the protein-encoding genes (1522 of 3079) could be assigned a putative function. An analysis of the secondary metabolite clusters revealed the presence of two uncharacterized phytoene synthase containing pathways and a novel siderophore pathway. Biological assays confirmed that the strain produces molecules with antioxidant and siderophore activities. Furthermore, intracellular extracts showed nematocidal activity towards C. elegans, suggesting that strain KRL4 is a source of anthelmintic compounds.

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“…For experiments shown in Table S1, B. thuringiensis 407 Δ spo0A :: kan cells transferred with pHY159 were grown in Luria broth overnight at 30°C in 10 µg/ml erythromycin to saturation. The next day, 100 µl of terpene or essential oil ( 80 ) was added to 100 µl of overnight culture to a final volume of 1 ml in Luria broth plus erythromycin (the following terpenes and essential oil used were: geraniol, eugenol, thymol, citral, carvacrol, cinnamic aldehyde, tea tree, limonene, and undecanoic acid). Tubes were shaken overnight at 30°C.…”

Section: Methodsmentioning

confidence: 99%

Recombinant Paraprobiotics as a New Paradigm for Treating Gastrointestinal Nematode Parasites of Humans

Abraham

Gazzola

et al. 2021

Antimicrob Agents Chemother

Self Cite

View full text Add to dashboard Cite

Gastrointestinal nematodes (GINs) of humans, e.g., hookworms, negatively impact childhood growth, cognition, nutrition, educational attainment, income, productivity, and pregnancy. Hundreds of millions of people are targeted with mass drug administration (MDA) of donated benzimidazole anthelmintics. However, benzimidazole efficacy against GINs is suboptimal, and reduced/low efficacy has been seen. Developing an anthelmintic for human MDA is daunting: it must be safe, effective, inexpensive, stable without a cold chain, and massively scalable. Bacillus thuringiensis (Bt) crystal protein 5B (Cry5B) has anthelmintic properties that could fill this void. Here we develop an Active Pharmaceutical Ingredient or API containing Bt Cry5B compatible with MDA. We expressed Cry5B in asporogenous Bt during vegetative phase, forming cytosolic crystals. These Bacteria with Cytosolic Crystals (BaCC) were rendered inviable (inactivated BaCC or IBaCC) with food-grade essential oils. IBaCC potency was validated in vitro against nematodes. IBaCC was also potent in vivo against human hookworm infections in hamsters. IBaCC production was successfully scaled to 350 liters at a contract manufacturing facility. A simple fit-for-purpose formulation to protect against stomach digestion and powdered IBaCC were successfully made and used against GINS in hamsters and mice. A pilot histopathology study and blood chemistry workup showed that five daily consecutive doses of 200 mg/kg Cry5B IBaCC (the curative single dose is 40 mg/kg) was non-toxic to hamsters and completely safe. IBaCC is a safe, inexpensive, highly effective, easy-to-manufacture, and scalable anthelmintic that is practical for MDA and represents a new paradigm for treating human GINs.

show abstract

Anthelmintic Activity of Yeast Particle-Encapsulated Terpenes

Cited by 21 publications

References 42 publications

Recombinant Paraprobiotics as a New Paradigm for Treating Gastrointestinal Nematode Parasites of Humans

Recombinant Paraprobiotics as a New Paradigm for Treating Gastrointestinal Nematode Parasites of Humans

Isolation and Characterization of Strain Exiguobacterium sp. KRL4, a Producer of Bioactive Secondary Metabolites from a Tibetan Glacier

Recombinant Paraprobiotics as a New Paradigm for Treating Gastrointestinal Nematode Parasites of Humans

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