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A Simple and Rapid Method for Blood Collection from Walking Catfish, Clarias Batrachus (Linneaus, 1758)

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A simple and rapid method for blood collection from walking catfish, Clarias batrachus (Linneaus, 1758) Item Type article Authors Argungu, L.A.; Siraj, S.S.; Christianus, A.; Amin, M.S.N.; Daud, S.K.; Abubakar, M.S.; Abubakar, I.A.; Aliyu-Paiko, M. Download date 01/10/2021 02:13:44 Link to Item http://hdl.handle.net/1834/37860 Iranian Journal of Fisheries Sciences 16(3) 935-944 2017 A simple and rapid method for blood collection from walking catfish, Clarias batrachus (Linneaus, 1758) Argungu L.A.¹; Siraj S.S.¹; Christianus A.1,4*; Amin M.S.N.1; Daud S.K.²; Abubakar M.S.3; Abubakar I.A. 3 ; Aliyu-Paiko M.4 Received: July 2015 Accepted: April 2016 Abstract Blood is collected from experimental animals for a wide range of scientific purposes including; hematology, clinical biochemistry parameters, immunology, studies in bacteriology, parasitology and investigations in reproductive performance and health. The number of methods employed to collect blood from fish include; the puncture of caudal vein, dorsal aorta or cardiac vessels and the severance of the caudal vein. Unfortunately, all these procedures are practically found to be slow and stressful to Clarias batrachus, including the popular caudal vein approach, likely due to the small size of caudal veins relative to the size of the species. In line with the universal ethical recommendations for taking blood from small research animals, we propose an alternative one-operator approach for C. batrachus that is simple, rapid and without the need to sacrifice the fish as with other methods. This procedure targets the dorsal aorta (a relatively larger blood vessel) in a sedated fish, punctured by inserting a needle directly from the anterior part of the anal fin about 2-5 mm behind the genital papilla, to Downloaded from jifro.ir at 16:46 +0330 on Sunday March 4th 2018 draw the desired amount of blood. The technique is a one-operator procedure not requiring the help of an assistant or any special equipment to restrain the fish. The operation of the protocol is unique since it permits the continuous collection of blood from the same experimental fish over a varied time course and reduces the need for a large number of replicate animals. The advantages of the proposed protocol are also highlighted and discussed in detail. Keywords: Haematology, Catfish, Aquaculture, Blood sampling, Clarias batrachus 1-Department of Aquaculture, Faculty of Agriculture, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia 2-Departments of Biology, Faculty of Science, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia 3-Department of Veterinary Pathology and Microbiology, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia 4-Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia *Corresponding author’s Email: [email protected] 936 Argungu et al., A simple and rapid method for blood collection from walking catfish, Clarias … Introduction available for blood collection from fish, Clarias batrachus is one of the most including the puncture of caudal vein, notable catfishes in Asia (Argungu et dorsal aorta or cardiac vessels, al., 2012) and its popularity has decapitation and the severance of increased significantly as an important caudal vein (MUAWC, 2008). commercial aquaculture species However, the choice of sampling (Zonneveld et al., 1988; Knud-Hansen technique utilized by a researcher et al., 1990; Kumari and Sahoo, 2006; depends on a number of factors such as Sahoo et al., 2007) and for aquarium the health status and size of fish, the trade in recent times among the fish quantity of blood required to be farming communities in Asia (Ng and collected and the fate of the animal Kottelat, 2008). This is better (terminal or non-lethal) in a study appreciated, considering that consumers (Svobodora and Vykusova, 1991). prefer it more, than Clarias gariepinus Overall, the chosen sampling approach in some parts of Asia because of its should be the least stressful, to avoid excellent taste and rough muscle mass. interference with the outcome of the This is in spite of the better growth study. This consideration makes it performance achieved with the latter necessary for the chosen methodology species when both are farmed (Rahman to be refined, in order to also comply et al., 1995). C. batrachus typically with International ethical and attains a standard length of 22.5-30 cm, procedural requirements, such as the although individuals twice this size has Australian Code of Practice for the been reported (Masterson, 2007). Care and Use of Animals for Scientific Lately, research efforts on C. batrachus Purposes (MUAWC, 2008). are focused on its reproductive The most popular technique, Downloaded from jifro.ir at 16:46 +0330 on Sunday March 4th 2018 performance, especially with regards to targeting the caudal vein to draw blood selection of matured candidates for (MUAWC, 2008) is practically slow induced breeding. To carry out such and stressful, particularly when applied studies effectively, the need arises for to C. batrachus. This is likely due to the researchers to collect blood from the small size of caudal veins in the experimental animals, for further species. Furthermore, the other processing (Kori-Slakpere, 1985). conventional methods are very Collection of blood from cumbersome and less convenient to experimental fish is for a wide range of sample blood in the species (via non- scientific purposes including lethal means). This is particularly a hematology, clinical biochemistry challenge in experiments which require parameters, studies in bacteriology, continuous blood collection over a parasitology and investigations in varied time course and need the animal reproductive performance and health to be kept alive. (DFO, 2004). A number of methods are Iranian Journal of Fisheries Sciences 16(3) 2017 937 It is an ethical obligation for researchers of clove oil solutions diluted in 5 L of to ensure that laboratory animals in water), for 4-6 min. Before the blood their care during experimentation are collection procedure, we subjected the not subjected to unnecessary stress fish samples to morphometric (DFO, 2004). Consistent with this measurements, to determine their body therefore, the objective of the present weight, as well as total and standard exposition was to develop a one- lengths. Complete sedation was operator approach that is simple and confirmed when a fish no longer rapid for sampling blood from C. responded to external stimuli, yet slight batrachus, without the need to sacrifice gill movements were still observable. the fish. The advantages of the Blood collection was accomplished proposed protocol are also highlighted as follows; the completely sedated fish and discussed in detail. was removed with the aid of a net from the anesthetic solution and placed on a Materials and methods clean towel on its side. Using the towel, The specimens we used to develop this the fish was carefully lifted to a protocol were C. batrachus brood fish, comfortable handling position with one with an average body weight and total hand, in such a way that the fish was length of 186 g and 28 cm, respectively. upside down and the head away from, The live fish were caught from the wild but tail nearer to the individual carrying by fishermen in Negeri Sembilan, out the procedure. With the other hand, Malaysia where they were purchased a 23G needle was inserted into the fish and transported to University Putra at a point anterior to the anal fin, and Malaysia Aquaculture hatchery unit, about 2-5 mm behind the genital Puchong-Selangor, Malaysia. The fish papilla. The insertion of the needle into Downloaded from jifro.ir at 16:46 +0330 on Sunday March 4th 2018 samples were kept in fiber glass tanks the fish was at a perpendicular angle to (2 tones capacity) for about two the ventral surface (at approximately months, under observation during 90º), until some blood entered into the acclimation in our research facility. The syringe or when the needle made average water quality conditions in the contact with the vertebral column (hard tanks during this period were impenetrable surface). Once the needle maintained at 26.5ºC, 5.7 ppm and 7.6 touched the vertebral column, it was for temperature, dissolved oxygen and withdrawn slightly, approximately 1 pH, respectively, and were confirmed mm, so that the blood vessel overlying optimal to maintain sound health of the the vertebral column could then be species. sampled easily and rapidly. This was Using an appropriate net, we done by gently pulling on the plunger randomly removed individual fish from by maintaining consistent pressure until the holding tank and immersed it into the desired quantity of blood is drawn anesthetic solution (containing 2.5 mL into the syringe. The collected blood 938 Argungu et al., A simple and rapid method for blood collection from walking catfish, Clarias … sample was slowly but quickly and biochemical studies. All the fish in transferred into collection tubes this experimental trial (ranging in size (containing EDTA or heparin or treated between 75 g and 310 g) made 100% as may be desired per individual recovery and survived, thereafter to experimental set up). The sampled fish later be subjected to further was then transferred to a recovery tank, experimentation. containing clean, aerated water (also prepared before the commencement of Discussion the procedure), where it recovered fully According to MUAWC (2008), the and was swimming freely within 45-60 most popular technique for blood minutes. sampling from fish is the caudal vein The tube containing sampled blood method. Attempts were made by other was gently mixed with the anti- workers using this popular method, to coagulant (as the case may be) by sample blood from over ten individuals carefully turning it upside down to mix of C. batrachus in our laboratory, but the contents. The tubes were then kept were met with little success.
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