title: Optimization and validation of Mycobacterium marinum-induced adult zebrafish model for evaluation of oral anti-tuberculosis drugs.
abstract: BACKGROUND
  Mycobacterium marinum has emerged as a suitable species for induction of tuberculosis-like disease in zebrafish, and various zebrafish models (larval and adult) for drug screening have been proposed in the literature. It is believed that an adult zebrafish model is more useful in drug screening because, apart from assessment of efficacy, one can obtain data on dosage, pharmacokinetics and overall health improvement. This study suggests a simple, cost-effective and resource-efficient protocol for screening of anti-tuberculosis drugs.
  
  
  METHODS
  The parameters used for assessment of infection as well as anti-bacterial response were: (a) bacterial count; and (b) body weight change. An optimization study was conducted to establish the concentration of bacteria required to produce a reproducible phenotype of tuberculosis (TB). A negative control (Amoxicillin) and anti-mycobacterial drugs (Isoniazid, Rifampicin, Moxifloxacin, Ethambutol and Isoniazid+Rifampicin) were used for validation of the protocol. All the drugs were administered orally.
  
  
  RESULTS
  An intra-peritoneal inoculation of 0.75million bacteria/fish was optimized for the model. All the anti-tuberculosis drugs showed efficacy in this model, whereas the negative control did not show any signs of reversing the parameters of M. marinum infection.
  
  
  CONCLUSIONS
  Adult zebrafish model of M. marinum-induced tuberculosis has not been fully exploited as a drug screening tool. In the present report, a protocol is suggested that is simple, reproducible and resource-efficient for screening of anti-tuberculosis agents. This protocol is an attempt to refine the published protocols and use this model as a surrogate model of human TB for the purpose of drug screening.
authors.affiliation: Drug Discovery Research Laboratory, Department of Pharmacy, Birla Institute of Technology & Science-Pilani, Hyderabad Campus, Hyderabad 500078, India.
authors.forename: Jonnalagadda Padma
authors.initials: JP
authors.lastname: Sridevi
authors.affiliation: Drug Discovery Research Laboratory, Department of Pharmacy, Birla Institute of Technology & Science-Pilani, Hyderabad Campus, Hyderabad 500078, India.
authors.forename: Hasitha Shilpa
authors.initials: HS
authors.lastname: Anantaraju
authors.affiliation: Drug Discovery Research Laboratory, Department of Pharmacy, Birla Institute of Technology & Science-Pilani, Hyderabad Campus, Hyderabad 500078, India; Dr. Reddy's Institute of Life Sciences, University of Hyderabad Campus, Gachibowli, Hyderabad 500046, India. Electronic address: pushkark@drils.org.
authors.forename: Pushkar
authors.initials: P
authors.lastname: Kulkarni
authors.affiliation: Drug Discovery Research Laboratory, Department of Pharmacy, Birla Institute of Technology & Science-Pilani, Hyderabad Campus, Hyderabad 500078, India.
authors.forename: Perumal
authors.initials: P
authors.lastname: Yogeeswari
authors.affiliation: Drug Discovery Research Laboratory, Department of Pharmacy, Birla Institute of Technology & Science-Pilani, Hyderabad Campus, Hyderabad 500078, India.
authors.forename: Dharmarajan
authors.initials: D
authors.lastname: Sriram
keywords: Adult zebrafish; Anti-tuberculosis drugs; Drug discovery; Mycobacterium marinum; Mycobacterium tuberculosis; Oral dosing