Comparative toxicity of lead (Pb2+), copper (Cu2+), and mixtures of lead and copper to zebrafish embryos on a microfluidic chip

Abstract

Investigations were conducted to determine acute effects of Pb 2þ and Cu 2þ presented individually and collectively on zebrafish embryos. Aquatic safety testing requires a cheap, fast, and highly efficient platform for real-time evaluation of single and mixture of metal toxicity. In this study, we have developed a microfluidic system for phenotype-based evaluation of toxic effects of Pb 2þ and Cu 2þ using zebrafish (Danio rerio) embryos. The microfluidic chip is composed of a disc-shaped concentration gradient generator and 24 culture chambers, which can generate one blank solution, seven mixture concentrations, and eight single concentrations for each metal solution, thus enabling the assessment of zebrafish embryos.To test the accuracy of this new chip platform, we have examined the toxicity and teratogenicity of Pb 2þ and Cu 2þ on embryos. The individual and combined impact of Pb 2þ and Cu 2þ on zebrafish embryonic development was quantitatively assessed by recording a series of physiological indicators, such as spontaneous motion at 22 hours post fertilization (hpf), mortality at 24 hpf, heartbeat and body length at 96 hpf, etc. It was found that Pb 2þ or Cu 2þ could induce deformity and cardiovascular toxicity in zebrafish embryos and the mixture could induce more severe toxicity. This chip is a multiplexed testing apparatus that allows for the examination of toxicity and teratogenicity for substances and it also can be used as a potentially costeffective and rapid aquatic safety assessment tool. V C 2015 AIP Publishing LLC.

Reduced metabolic rate and increased heart beat as early signs of sub lethal Copper toxicity in developing Zebrafish

“…A significant observed effect of Cu was the morphological abnormalities, severe pericardial edema, spine curvature, posterior body scoliosis, caudal tip malformation, and abnormal fattening of the posterior part in zebrafish embryos. These results are in agreement with reported pathological problems after exposure to Cu, such as teratogenicity of Pb 2+ and Cu 2+ on zebrafish embryos, including bent spine, double tail spine, pericardial edema, and yolk sac edema, while several kinds of malformations often appeared together in one larva (Li et al, 2015;Sfakianakis et al, 2015;Zhang et al, 2015;Almond and Trombetta, 2016;Ganesan et al, 2016). Exposure to Cu significantly affected embryonic development, with groups exposed to Cu, exhibiting lower rates of utilisation of yolk sac nutrient reserves.…”

A millifluidic chip for cultivation of fish embryos and toxicity testing fabricated by 3D printing technology

“…not negatively inuence the development of the cultivated embryos in the designed chip. This was in agreement with the results in other systems, 21,23 where a normal embryo development was observed in long-term experiments under ow rates in range from 4 ml min À1 to 100 ml min À1 .…”

“…It can be e.g. a continuous 20,23 or a time-dependent sequential ow 26 of the media or toxicant. Another possibility is a sequential ow switched between a continuous ow of the media and the toxicant, 21 or a looping of the ow 26,39 of the toxicant in the cultivation chamber with its possible regular exchange.…”

“…The current microuidics benets from the use of polymers as building materials mainly due to their low costs and simplied manufacturing procedures. 27 The most published techniques employed for the zebrash embryo cultivation system are techniques using PDMS, where the master was manufactured by laser cutting 23,25 or by 3D printing. 28 Also, individual PMMA sheets cut by a laser micromachining method can be layered and assembled by thermal bonding to create a suitable cultivation microenvironment.…”

Changes in Arsenic and Copper Bioavailability and Oxytetracycline Degradation during the Composting Process

“…Copper addition increases the copper concentration in animal waste and manure [17,18]. Copper and other heavy metals show antimicrobial activity and are toxic toward embryos of Gobiocypris rarus and zebrafish [19,20,21]. Excessive Cu can also harm livestock and humans [22].…”