Repeated Blood Collection for Blood Tests in Adult Zebrafish

Abstract

Repeated blood collection is one of the most common techniques performed on laboratory animals. However, a non-lethal protocol for blood collection from zebrafish has not been established. The previous methods for blood collection from zebrafish are lethal, such as lateral incision, decapitation and tail ablation. Thus we have developed a novel "repeated" blood collection method, and present here a detailed protocol outlining this procedure. This method is minimally invasive and results in a very low mortality rate (2.3%) for zebrafish, thus enabling repeated blood sampling from the same individual. The maximum volume of blood sampling is dependent on body weight of the fish. The volume for repeated blood sampling at intervals should be ≤0.4% of body weight every week or ≤1% every 2 weeks, which were evaluated by measurements of blood hemoglobin. Additionally, hemoglobin, fasting blood glucose, plasma triacylglycerol (TG) and total cholesterol levels in male and female adult zebrafish were measured. We also applied this method to investigate the dysregulation of glucose metabolism in diet-induced obesity. This blood collection method will allow many applications, including glucose and lipid metabolism and hematological studies, which will increase the use of zebrafish as a human disease model organism. Video LinkThe video component of this article can be found at

Metabolomics and fish nutrition: a review in the context of sustainable feed development

“…The volume of blood samples is another issue if a noninvasive sampling method is set up. Approximately 0.5% of total body weight (i.e., 0.5 ml for a 100 g fish) can be collected without or with low mortality (Iwama et al 2011;Zang et al 2015). In general, metabolomic approaches use millilitre samples of blood although microlitre samples may suffice depending on the techniques employed or the instrumental setup.…”

“…; Zang et al . ). In general, metabolomic approaches use millilitre samples of blood although microlitre samples may suffice depending on the techniques employed or the instrumental setup.…”

The presence of a conspecific induces risk‐taking behaviour and enlargement of somata size of dopaminergic neurons in the brain of male medaka fish

“…Immediately after blood collection using a heparinized capillary glass, the concentration of blood glucose was measured using a commercial hand-held glucometer (Glutest neo alpha, GT-1830; PHC Co., Tokyo, Japan), as described by Zang et al (2015).…”

Comment: Practices for Drawing Blood Samples from Teleost Fish

“…), including repeated samples (Zang et al. ). Some analytical techniques (e.g., transcriptomics and endocrinology) require relatively small quantities of blood for analysis such that repeated blood sampling is possible even from fish of small body sizes (Lewis et al.…”

“…Despite Duman et al (2019) suggesting that blood sampling is restricted to "large" fish, researchers have successfully drawn blood from even tiny fish, such as Zebrafish Danio rerio (Pedroso et al 2012), including repeated samples (Zang et al 2015). Some analytical techniques (e.g., transcriptomics and endocrinology) require relatively small quantities of blood for analysis such that repeated blood sampling is possible even from fish of small body sizes (Lewis et al 2010;Cook et al 2012).…”