Alternative Methods in Biological Assays

A special issue of Animals (ISSN 2076-2615). This special issue belongs to the section "Animal Welfare".

Deadline for manuscript submissions: closed (31 December 2020) | Viewed by 10278

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Special Issue Information

Dear Colleagues,

Experimental animal data are the cornerstone in pharmacology, toxicology, and risk assessment; however, promoting good animal welfare is a contentious issue in drug research. The need to reduce the pain and distress experienced by animals during scientific experiments highlights the importance of the principles of replacing, reducing, and refining animal use in the study of biomolecules. There is an urgent need to develop and use replacement and reduction alternative techniques that could substitute for a significant proportion of animal research at every level of preclinical studies.

Several alternative models, such as copepod (Tigriopus spp.), water flea (Daphnia spp.), brine shrimp (Artemia salina), Caenorhabditis elegans, fathead minnow (Pimephales promelas), zebrafish (Danio rerio), and rainbow trout (Oncorhynchus mykiss), have been developed and proven to be successful, especially with respect to the testing of chemical toxicity. Some of these models could be extended in many other biomedical research fields, including basic developmental biology or pharmacology, in order to capitalize on their potential to model human diseases. Lower vertebrates are an attractive option because of the reduced ethical impact and their good genetic relatedness to mammals.

This Special Issue will focus on research studies highlighting the development of alternative assays and testing strategies to model human responses. This Special Issue aims to bring together multi- and interdisciplinary approaches driven by the concept of the 3Rs (reduction, refinement, and replacement) in drug development. Of special interest is research on the use of alternative species or methods for complex pharmacology studies, such as those related to memory, aging, endocrine dysfunction, muscle dystrophy, and diabetes, to name a few.

Prof. Dr. Nitulescu George Mihai
Guest Editor

Manuscript Submission Information

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Keywords

  • alternative pharmacology tests
  • 3Rs
  • Daphnia spp.
  • Artemia salina
  • Danio rerio
  • Caenorhabditis elegans

Published Papers (3 papers)

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Research

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18 pages, 1984 KiB  
Article
Toxicity of Carbon, Silicon, and Metal-Based Nanoparticles to the Hemocytes of Three Marine Bivalves
by Konstantin Pikula, Vladimir Chaika, Alexander Zakharenko, Anastasia Savelyeva, Irina Kirsanova, Anna Anisimova and Kirill Golokhvast
Animals 2020, 10(5), 827; https://doi.org/10.3390/ani10050827 - 10 May 2020
Cited by 18 | Viewed by 2870
Abstract
Nanoparticles (NPs) have broad applications in medicine, cosmetics, optics, catalysis, environmental purification, and other areas nowadays. With increasing annual production of NPs, the risks of their harmful influence on the environment and human health are also increasing. Currently, our knowledge about the mechanisms [...] Read more.
Nanoparticles (NPs) have broad applications in medicine, cosmetics, optics, catalysis, environmental purification, and other areas nowadays. With increasing annual production of NPs, the risks of their harmful influence on the environment and human health are also increasing. Currently, our knowledge about the mechanisms of the interaction between NPs and living organisms is limited. The marine species and their habitat environment are under continuous stress owing to the anthropogenic activities, which result in the release of NPs in the aquatic environment. We used a bioassay model with hemocytes of three bivalve mollusc species, namely, Crenomytilus grayanus, Modiolus modiolus, and Arca boucardi, to evaluate the toxicity of 10 different types of NPs. Specifically, we compared the cytotoxic effects and cell-membrane polarization changes in the hemocytes exposed to carbon nanotubes, carbon nanofibers, silicon nanotubes, cadmium and zinc sulfides, Au-NPs, and TiO2 NPs. Viability and the changes in hemocyte membrane polarization were measured by the flow cytometry method. The highest aquatic toxicity was registered for metal-based NPs, which caused cytotoxicity to the hemocytes of all the studied bivalve species. Our results also highlighted different sensitivities of the used tested mollusc species to specific NPs. Full article
(This article belongs to the Special Issue Alternative Methods in Biological Assays)
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15 pages, 2437 KiB  
Article
Metabolomic Profile of Primary Turkey and Rat Hepatocytes and Two Cell Lines after Chloramphenicol Exposure
by Lidia Radko, Tomasz Śniegocki, Bartosz Sell and Andrzej Posyniak
Animals 2020, 10(1), 30; https://doi.org/10.3390/ani10010030 - 21 Dec 2019
Cited by 6 | Viewed by 2759
Abstract
The purpose of this study was to assess the formation of chloramphenicol metabolites in primary turkey and rat hepatocyte cultures and human hepatoma (HepG2) cells and nonhepatic, Balb/c 3T3 fibroblasts. Additionally, the cytotoxicity of the drug was assessed through three biochemical endpoints: mitochondrial [...] Read more.
The purpose of this study was to assess the formation of chloramphenicol metabolites in primary turkey and rat hepatocyte cultures and human hepatoma (HepG2) cells and nonhepatic, Balb/c 3T3 fibroblasts. Additionally, the cytotoxicity of the drug was assessed through three biochemical endpoints: mitochondrial and lysosomal activity and cellular membrane integrity after 24 and 48 h exposure. The two metabolites of the drug, chloramphenicol glucuronide and nitroso-chloramphenicol, were detected to the greatest extent in both primary hepatocyte cultures by liquid chromatography–tandem mass spectrometry. Toxic nitroso-chloramphenicol was the main metabolite in the primary turkey hepatocyte cultures, but it was not in the primary rat hepatocyte cultures. The most affected endpoint in turkey and rat hepatocyte cultures was the disintegration of the cellular membrane, but in the cell lines, mitochondrial and lysosomal activities underwent the greatest change. The primary hepatocyte cultures represent valuable tools with which to study the species differences in the biotransformation and toxicity of drugs. To the best of our knowledge, this is the first report of differences in chloramphenicol metabolism in primary turkey and rat hepatocyte cultures. Full article
(This article belongs to the Special Issue Alternative Methods in Biological Assays)
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Review

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21 pages, 575 KiB  
Review
Wild-Type Zebrafish (Danio rerio) Larvae as a Vertebrate Model for Diabetes and Comorbidities: A Review
by Maryna van de Venter, Jenske Didloff, Shanika Reddy, Bresler Swanepoel, Sharlene Govender, Ntokozo Shirley Dambuza, Saralene Williams, Trevor Craig Koekemoer and Luanne Venables
Animals 2021, 11(1), 54; https://doi.org/10.3390/ani11010054 - 30 Dec 2020
Cited by 4 | Viewed by 4080
Abstract
Zebrafish have become a popular alternative to higher animals in biomedical and pharmaceutical research. The development of stable mutant lines to model target specific aspects of many diseases, including diabetes, is well reported. However, these mutant lines are much more costly and challenging [...] Read more.
Zebrafish have become a popular alternative to higher animals in biomedical and pharmaceutical research. The development of stable mutant lines to model target specific aspects of many diseases, including diabetes, is well reported. However, these mutant lines are much more costly and challenging to maintain than wild-type zebrafish and are simply not an option for many research facilities. As an alternative to address the disadvantages of advanced mutant lines, wild-type larvae may represent a suitable option. In this review, we evaluate organ development in zebrafish larvae and discuss established methods that use wild-type zebrafish larvae up to seven days post fertilization to test for potential drug candidates for diabetes and its commonly associated conditions of oxidative stress and inflammation. This provides an up to date overview of the relevance of wild-type zebrafish larvae as a vertebrate antidiabetic model and confidence as an alternative tool for preclinical studies. We highlight the advantages and disadvantages of established methods and suggest recommendations for future developments to promote the use of zebrafish, specifically larvae, rather than higher animals in the early phase of antidiabetic drug discovery. Full article
(This article belongs to the Special Issue Alternative Methods in Biological Assays)
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