ABSTRACT
Lyme disease is on the rise. Caused by a spirochete Borreliella burgdorferi, it affects an estimated 500,000 people in the United States alone. The antibiotics currently used to treat Lyme disease are broad spectrum, damage the microbiome, and select for resistance in non-target bacteria. We therefore sought to identify a compound acting selectively against B. burgdorferi. A screen of soil micro-organisms revealed a compound highly selective against spirochetes, including B. burgdorferi. Unexpectedly, this compound was determined to be hygromycin A, a known antimicrobial produced by Streptomyces hygroscopicus. Hygromycin A targets the ribosomes and is taken up by B. burgdorferi, explaining its selectivity. Hygromycin A cleared the B. burgdorferi infection in mice, including animals that ingested the compound in a bait, and was less disruptive to the fecal microbiome than clinically relevant antibiotics. This selective antibiotic holds the promise of providing a better therapeutic for Lyme disease and eradicating it in the environment.
Subject(s)
Anti-Bacterial Agents/therapeutic use , Lyme Disease/drug therapy , Animals , Borrelia burgdorferi/drug effects , Calibration , Cinnamates/chemistry , Cinnamates/pharmacology , Cinnamates/therapeutic use , Drug Evaluation, Preclinical , Feces/microbiology , Female , HEK293 Cells , Hep G2 Cells , Humans , Hygromycin B/analogs & derivatives , Hygromycin B/chemistry , Hygromycin B/pharmacology , Hygromycin B/therapeutic use , Lyme Disease/microbiology , Mice , Microbial Sensitivity Tests , Microbiota/drug effectsABSTRACT
During the construction of a physical map for Leishmania major (LV39) chromosome 2 we have rescued and characterized a L. major (LV39) derived genomic clone bearing solely as insert a long stretch of the miniexon gene array. The recombinant was devised as a tool to study the effect of miniexon overexpression on virulence and growth advantage. Such clone, 32D05, contains approximately 40 kb of the miniexon tandem array. We have examined the course of infection in susceptible BALB/c mice inoculated with transfectants carrying 32D05 as an episome. The study was carried out in two different clonal lines of L. major: virulent line LV39 (clone 5) and avirulent LT252 (CC1 clone). The results presented here indicate that high levels of miniexon expression affect negatively the ability of once virulent lines to induce lesions when injected in susceptible mice.
Subject(s)
Exons/genetics , Gene Expression Regulation , Genes, Protozoan , Leishmania major/genetics , Leishmania major/pathogenicity , Animals , Anthelmintics/pharmacology , Clone Cells , Exons/drug effects , Female , Gene Dosage , Gene Expression Regulation/drug effects , Genes, Protozoan/drug effects , Genome, Protozoan , Hygromycin B/therapeutic use , Leishmania major/drug effects , Leishmaniasis, Cutaneous/drug therapy , Leishmaniasis, Cutaneous/genetics , Leishmaniasis, Cutaneous/parasitology , Mice , Mice, Inbred BALB C , Mice, Inbred C3H , RNA, Protozoan/analysis , TransfectionABSTRACT
Hepatitis caused by mouse hepatitis virus (MHV-A59), a murine coronavirus, is accompanied by direct infection and replication of virus within the liver. We demonstrate here that the aminoglycoside hygromycin B is able to eliminate MHV-A59 infection from mouse peritoneal macrophages and cultured liver cells in vitro and is also able to reduce levels of virus replication and necrotic liver foci in vivo.
Subject(s)
Hepatitis, Viral, Animal/drug therapy , Hygromycin B/therapeutic use , Murine hepatitis virus , Animals , Cell Survival/drug effects , Cells, Cultured , Fluorescent Antibody Technique , Hepatitis, Viral, Animal/microbiology , Hygromycin B/toxicity , Liver/microbiology , Macrophages/drug effects , Macrophages/microbiology , Mice , Mice, Inbred A , Mice, Inbred BALB C , Virus Replication/drug effectsABSTRACT
The relatively cell impermeable hygromycin B was found to inhibit viral but not cellular protein synthesis when added to cultures of murine hepatitis virus (MHV)-infected or mock-infected mouse L-2 fibroblasts. Membrane permeability, as judged by influx of sodium ions, has previously been demonstrated to be an MHV E2 glycoprotein-mediated, cytopathic effect of MHV infection in L-2 cells. It is therefore likely that the selective effect of hygromycin B on viral protein synthesis is a reflection of an increased drug penetration into virus-infected cells. Using hygromycin B as a marker for MHV-induced cell membrane cytopathology, the effects of drug treatment on a persistent MHV infection in mouse LM-K fibroblasts were investigated. MHV persistence in LM-K cells, which normally involves a steady state infection of 0.1 to 1% of the cells in culture, was found to be cured by hygromycin B treatment, as measured by the elimination of infectious virus from the supernatant medium. Hygromycin B also resulted in the eradication of MHV-specific RNA from LM-K cells, arguing against the presence of a non-cytopathically or latently infected subpopulation of cells.
Subject(s)
Coronaviridae Infections/etiology , Disease Models, Animal , Animals , Coronaviridae Infections/drug therapy , Coronaviridae Infections/microbiology , Cytopathogenic Effect, Viral/drug effects , Dose-Response Relationship, Drug , Hepatitis, Viral, Animal/drug therapy , Hepatitis, Viral, Animal/etiology , Hepatitis, Viral, Animal/microbiology , Hygromycin B/therapeutic use , L Cells , Mice , Murine hepatitis virus/drug effects , Murine hepatitis virus/metabolism , Viral Proteins/antagonists & inhibitors , Viral Proteins/biosynthesisABSTRACT
In vitro and in vivo screening methods were established for the discovery of new active substances against Treponema hyodysenteriae. During the screening methods, hygromycin A produced by Streptomyces hygroscopicus KA-355 was found to be active against T. hyodysenteriae. Hygromycin A did not show high antitreponemal activity in in vitro test using the paper disc method on the agar plate inoculated with T. hyodysenteriae. However, the antibiotic exhibited highly therapeutic effect in CF-1 mice, compared with of lincomycin, tiamulin, lankacidin C or olaquindox drinking water. The effective dose (ED50) of hygromycin A was 1.1 micrograms/ml.
Subject(s)
Anti-Bacterial Agents/therapeutic use , Cinnamates , Hygromycin B/therapeutic use , Treponemal Infections/drug therapy , Administration, Oral , Animals , Drug Evaluation, Preclinical , Female , Hygromycin B/analogs & derivatives , Mice , Mice, Inbred Strains , Species SpecificityABSTRACT
This study was conducted to evaluate hygromycin A fed to growing swine at 1, 5, 10 or 20 g/ton feed for the control of Treponema hyodysenteriae-caused dysentery. Pigs provided carbadox at 50 g/ton feed served as an infected treatment control group. All pigs were orally, via stomach intubation, administered 100 ml of a T. hyodysenteriae broth culture. During the in vivo test, rectal swabs were taken for T. hyodysenteriae isolation, body weights of all pigs and the feed consumption was determined. All pigs were euthanized and necropsied at study end; the large intestine was cultured for T. hyodysenteriae and gross intestinal lesions were noted. T. hyodysenteriae-caused swine dysentery was successfully controlled by feeding hygromycin A at 5 g/ton. Hygromycin A medicated pigs performed as well as or better than carbadox-medicated pigs.
