Rederivation of a mutant line (prop 1) of zebrafish Danio rerio infected with Pseudoloma neurophilia using in vitro fertilization with eggs from pathogen‐free wild‐type (AB) females and sperm from prop 1 males

Along with the growing number of laboratories that work with zebrafish (Danio rerio), it is necessary to have animals with good sanitary quality. Specific pathogens can interfere with the experimental results and in the life quality of the animals. Pseudoloma neurophilia is a parasite with high pote...

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Published inJournal of fish diseases Vol. 45; no. 1; pp. 35 - 39
Main Authors Ventura Fernandes, Bianca H., Caetano da Silva, Caroline, Bissegato, Debora, Kent, Michael L., Carvalho, Luciani R.
Format Journal Article
LanguageEnglish
Published England Blackwell Publishing Ltd 01.01.2022
Subjects
Animals
Biological fertilization
Danio rerio
Disinfection
Eggs
Female
Fertilization in Vitro
Fish
Fish Diseases
Freshwater fishes
Genetic modification
Heterozygotes
In vitro fertilization
Laboratories
Male
Microsporidia
Microsporidiosis
Mutants
Nucleotide sequence
Parasites
pathogen
Pathogens
PCR
Pseudoloma neurophilia
Quality of life
Spermatozoa
SPF
Zebrafish
SPF
Pseudoloma neurophilia
pathogen
zebrafish
Danio rerio
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Summary:Along with the growing number of laboratories that work with zebrafish (Danio rerio), it is necessary to have animals with good sanitary quality. Specific pathogens can interfere with the experimental results and in the life quality of the animals. Pseudoloma neurophilia is a parasite with high potential for interference in behavioural, morphology, toxicological and genetic research, and is very common in zebrafish facilities. With that, we implemented a protocol for the pathogen elimination in a genetically modified lineage (prop 1) using eggs from specific pathogen‐free (SPF) wild‐type fish (AB line) for in vitro fertilization, along with water recirculation equipment disinfection, appropriate PCR screening and back crossing protocols. This resulted in SPF prop 1 heterozygotes, which allowed us to move forward with subsequent crossings to develop homozygote prop 1 mutants for our research. Hence, this demonstrates a useful strategy for an individual research laboratory to rederive a specific mutant free line that is not available from other SPF laboratories.
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content type line 23
ISSN:0140-7775
1365-2761
DOI:10.1111/jfd.13529