Effect of Storage Temperature on African Catfish (Clarias Gariepinus Burchell 1822) Milt Quality

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1 International Journal of Innovative Studies in Aquati Biology and Fisheries Volume 4, Issue 1, 2018, PP 7-12 ISSN (Print) & ISSN (Online) DOI: Effet of Storage Temperature on Afrian Catfish (Clarias Gariepinus Burhell 1822) Milt Quality Idahor K. O. 1, Okunseor S. A. 2, Sokuni O. A. 3, Osaiyuwu O. H. 3, Osayande U. D. 3, Hamza J. 1, Isah N. 2 1 Department of Animal Siene, Nasarawa State University, Keffi, Shau-Lafia Campus, P.M.B. 135, Lafia, , Nigeria. 2 Department of Aquaulture and Fisheries Management, Nasarawa State University, Keffi, Shau -Lafia Campus, P.M.B. 135, Lafia, , Nigeria. 3 Animal Siene Department, Faulty of Agriulture and Forestry, University of Iadan, Iadan, Nigeria. *Corresponding Author: Idahor K. O, Department of Animal Siene, Nasarawa State University, Keffi, Shau-Lafia Campus, P.M.B. 135, Lafia, , Nigeria. Astrat: Afrian atfish (C. gariepinus) does not reprodue in aptive, hene the adoption of artifiial reeding strategy. This study was targeted at the possiility of storing milt at different temperatures, to ensure all-year-round reeding operation. To aomplish this aim, milt was harvested from a total of twelve Afrian atfish and stored at three different temperatures designated as treatments T1 (26.3 o C): Benh temperature (ontrol: without refrigeration), T2 (7.42 o C): Fridge temperature and T3 (1.3 o C): Freeze temperature. Results indiated that the room and testes temperature values were onstantly 36.4 o C and C, standard ody and total ody length values ranged from 42 to 43 m and 48 to 50 m respetively. While the papilla length value varied from 1.5 to 1.8 m, the fin length value was etween 8.5 and 13.3 m. The sperm ells onentration value was as high as x 10 9 aross the treatments and the alive sperm ells were up to 75 79% at 0 minute ut deteriorated to as low as 1.5 4% at 15 minutes. Though some alive and motile sperm ells were oserved at 1 hour, the perentage was as low as 0.25%, suggesting that atfish milt stored without ryopreservation might not e viale in artifiial reeding. Keywords: Afrian atfish, artifiial reeding, milt storage, refrigeration, 1. INTRODUCTION Afrian atfish is elieved to e native of Afria with a wide geographial distriution within the ontinent. It elongs to the family of Claridae onsisting of aout 14 genera, 5 sugenera and 32 speies inluding Clarias, Clarioides, Anguillo, Platyephaloides and Breviephaloides [1, 2, 3]. Clarias gariepinus is one of the most suitale speies for aquaulture in Afria attaining aout a kg ody weight at aout 6 months of age espeially under intensive management onditions. It was reported that C. gariepinus was hardy and adapted to diverse environments even with poor water quality [4]. It is highly herished y many Nigerians, hene it ommands very high ommerial value and it is the most popular ultured fish in the ountry though still at susistene level. Unfortunately, C. gariepinus does not reed in aptive hene, some artifiial reeding approahes [5, 6 and 7]. Meanwhile, C. gariepinus eggs, larvae, fry and fingerlings ould e olleted from natural environment for stoking ut may not e availale all-year-round [8, 9, 10, and 11]. Thus, it was suggested [12] that systemati pond propagation method would ensure fish prodution all-year-round. In this method, the female is injeted with an appropriate hormone to failitate ooyte development and egg maturation prior to spawning. Regrettaly, the male must e killed to otain the milt resulting in sarity of male rood stok. In some ases, the milt is harvested from many males depending on the ommerial status of the farm yet, either the harvested milt is not enough warranting killing of more males or is in exess of what is needed. This often led to olossal losses. At the moment, there is little or no information on alternative means of reduing this olossal wastage exept, the use of syringe attempted y [13], ryopreservation tehnique propounded y [14] and International Journal of Innovative Studies in Aquati Biology and Fisheries (IJISABF) Page 7

2 Effet of Storage Temperature on Afrian Catfish (Clarias Gariepinus Burhell 1822) Milt Quality simulation of natural onditions with little or no onsiderale natural spawning tendeny [5, 6, 15, 16]. Sine all these proedures neessitated the use of suitale reagents and high tehnial-knowhow, it may not e pratiale at the farmstead level in Nigeria. Moreover, milt quality is measured with the poteny or aility to suessfully fuse with the stripped eggs. This may not e ahievale if the harvested milt is kept longer than neessary outside the fish. Essentially, spermatozoa fertilizing aility depends on omposition of seminal fluid, milt volume, spermatozoa density and motility [17, 18, 19]. Most ommerial hatheries where Afrian atfish seeds are propagated, inadequate quantity and quality of milt always result in reeding failure [20]. Consequently, this study was aimed at the possiility of storing C. gariepinus milt at different temperature for susequent utilization in artifiial insemination. This may possily ensure all-year-round reeding plan in fish as otainale in most livestok artifiial insemination programs. 2. MATERIALS AND METHODS 2.1. Experimental Design and Fish Welfare Twelve male C. gariepinus rood stok weighing etween and 0.66kg were ought from a sales outlet in Bukan Sidi, Lafia and were transported in jerry an ontaining lean water to Animal Siene Laoratory, Faulty of Agriulture, Nasarawa State University, Keffi, Shau-Lafia Campus. The fish were assigned to three treatments in a ompletely randomised design ased on live weight. The treatments were Benh temperature (T1) (ontrol: without refrigeration), Fridge temperature (T2) and Freeze temperature (T3) and eah treatment had four repliates. This experiment was arried out purely as an oservational work so, ethial permission was not onsidered neessary Data Colletion and Analysis Immediately on arrival, the twelve fish were transferred into an open owl ontaining lean water where eah of them was piked and weighed using tale sale (Five Goats ) to otain live weight. The standard ody length was taken from the mouth to the audal fin ase, while the total ody length was taken from the mouth to the end of the audal fin using measuring tape (Butterfly ). Papilla length, left and right fin length were measured using measuring tape (Butterfly ). Thereafter, all the fish were sarified y perussive stunning the head aove the eyes to impat on the rain with a lunt ojet until the fish was unonsious. The adomen was arefully ut open and thermometer was plaed on the testes diretly to otain testes temperature. Another thermometer was hung on the wall to otain room temperature and a separate one was used in monitoring the refrigerator temperature. The left and right testes were olleted arefully and the length was taken using measuring tape (Butterfly ). The testes were arefully inised using surgial lade and the milt was gently squeezed out into Eppendorf tue, to measure the volume and the milt ph was determined with a ph strip. The harvested milt from the four repliates in eah treatment was pooled, stirred gently with a spatula and stored on the Benh, in the Fridge and Freezer temperature respetively. The pooled milt was proessed for quality evaluation aording to the proedures of [21] and [22]. Aliquot of the proessed milt was taken with a miropipette and a drop was olleted with a mirosope slide, overed with a slip and viewed in a mirosope (Olympus Mirosope Tokyo, Japan) at 10x and 40x magnifiations, to determine alive and dead spermatozoa ells, ative and sluggish spermatozoa ells as well as spermatozoa ells onentration as reported [23]. The pooled milt was always returned to the appropriate position promptly and this proedure was repeated every 15 minutes until no more motile spermatozoon was oserved. Meanwhile, values of the milt volume and sperm ells onentration were determined only at 0 minute. All the sets of data olleted were sujeted to analysis of variane aording to statistial proedure of [24] and where appliale, the mean values were separated ased on Dunan Multiple Range Test of the same software pakage. 3. RESULTS Tale 1 presents the temperature and ody linear parameters of the experimental atfish. There were statistial differenes (P<5) in all the parameters measured exept in room and testes temperature values. The milt storage temperature value was oolest (1.3 0 C) in T 3 followed y T 2 ( C) and T 1 ( C) in that arrangement. The live weight value ranged from to 0.64kg, standard ody length International Journal of Innovative Studies in Aquati Biology and Fisheries (IJISABF) Page 8

3 Effet of Storage Temperature on Afrian Catfish (Clarias Gariepinus Burhell 1822) Milt Quality (42.4 to 43.1 m), total ody length (48.2 to 50.2 m) and the papilla length varied from 1.5m in T 2 to 1.8m in T 1. The left fin length was as short as 8.5 m in T 2 ompared to 13.3 m reorded in T 3. Whereas, the right fin length was not signifiantly different (P>5) in oth T 1 and T 3 ut the mean values were statistially different (P<5) from 13.3 m reorded in T 2. While the left testis length mean value ranged from 4.69 m in T 1 to 5.5 m in T 3, the right testis was as long as 6.1 m in T 2. Tale1. Temperature parameters and ody linear measurements of the experimental atfish Treatments (STD) Parameters T1 T2 T3 Room Temperature ( 0 C) Testis temperature ( 0 C) Stored milt temperature ( 0 C) 26.3 a Live weight (kg) a 1 08 Standard ody length (m) Total ody length (m) a Papilla length (m) a Left fin length (m) a a Right fin length (m) a Left testis length (m) a 5.58 a Right testis length (m) a 5.68 a,, : Means with different letters on the same row are signifiantly different at P<5; STD: Plus or minus standard deviation; T1: Benh temperature (ontrol: without refrigeration); T2: Fridge temperature; T3: Freeze temperature. The effet of different storage time interval on Afrian atfish milt quality is shown in Tale 2. There were signifiant differenes (P<5) in all the parameters monitored aross the treatments exept at 0 minute, where the alive sperm ells, dead sperm ells, ative motile sperm ells and sluggish motile sperm ells values did not differ statistially (P>5). The milt volume mean value was highest ( ml) in T 2 followed y T 1 (0.5 ml) and T 3 where 0.4 ml was reorded. It was oserved that milt ph mean values whih ranged from 6.85 to 8.9, were not persistently different (P>5) aross the treatments in T 2 and T 3, ut oth values differed signifiantly (P<5) from that of T3. At 0 minute, the values of alive sperm ells, dead sperm ells, ative motile sperm ells and sluggish motile sperm ells varied etween 75 and 79%, 21 and 25%, 72 and 76% as well as 24 and 28% in that order. Whereas, at 15 minutes, the values deteriorated to etween 1.5 and 4% alive sperm ells, 96 and 98.5% dead sperm ells, 7.25 and 17.5% ative motile sperm ells and sluggish sperm ells varied from 82.5 to 92.8%. At 30 minutes and eyond, asolutely % dead sperm ells were reorded exept at 1 hour, where 0.25% of the Freeze milt (T 3 ) was alive and out of this value, only 24% was atively motile while 76% was sluggish. The sperm ells onentration value whih varied from x 10 9 aross the treatments, was onsistently superior in T 2 followed y T 1 and T 3 respetively. Tale2. Effet of different storage time interval on Afrian atfish milt quality Para mete rs ᶲMV (ml) Milt ph ASC DSC Treatments (STD) hour 1:15 hour 1 minutes 45 minutes 30 minutes 15 minute 0 T1 T2 T3 T1 T2 T3 T1 T2 T3 T1 T2 T3 T1 T2 T3 T1 T2 T a a a a a a a a a a a a International Journal of Innovative Studies in Aquati Biology and Fisheries (IJISABF) Page 9

4 Effet of Storage Temperature on Afrian Catfish (Clarias Gariepinus Burhell 1822) Milt Quality a a AMS C SMS C ᶲSCC (x10 9 ) ᶲMilt volume and sperm ells onentration were determined only at 0 minute; Mean values omparison was per time treatment only and not aross treatments; a,, : Means with different letters on the same row at the same time per treatment are signifiantly different at P<5; STD: Plus or minus Standard deviation; T 1 (26.3 o C): Benh temperature (ontrol: without refrigeration); T 2 (7.42 o C): Fridge temperature; T 3 (1.3 o C): Freeze temperature; MV: Milt volume; ASC: Alive sperm ells; DSC: Dead sperm ells; AMSC: Ative motile sperm ells; SMSC: Sluggish motile sperm ells; SCC: Sperm ells onentration. 4. DISCUSSION The results indiated that the milt storage temperatures varied onsideraly and the values were within 1 to 7 0 C desried as preservation and storage temperature [25]. It was shown that the ody linear parameters were similar to those reported in Afrian mudfish aught in a dam [26]. However, the live weight value was lower than kg reported [27] in aptive Afrian mudfish at 8 months old yet, the standard ody and total ody length values were similar. This oservation proaly showed that at maturity, the ody linear values may not inrease irrespetive of the final ody weight attained, suggesting that the experimental fish were perhaps sexually matured. The high sperm ells onentration reorded in this study, ompared favouraly well with 5.16 x10 8 reported in C. gariepinus [28] thus further strengthen the assertion that the experimental fish were perhaps sexually matured. It was oserved that at 0 minute, the milt quality evaluation was est and at 1 hour however, some alive and motile sperm ells were oserved ut the motility level was so low that it may proaly have little or no fertilizing aility if used in spawning aording to several reports [18, 19, and 29]. This possily suggested that refrigeration storage of milt without ryopreservation materials may not e suitale in artifiial insemination in aptive atfish prodution. This oservation apparently validated several reports, on the need for ryopreservation in artifiial reeding operation in Afrian atfish [14, 20, 28, and 30]. 5. CONCLUSION The ody linear measurements indiated that the experimental fish were sexually matured with very high sperm ells onentration, indiative of high potential fertilizing aility if used in spawning immediately or if properly preserved and stored. It was however revealed that Fridge and Freeze temperatures ould not prevent the degenerative iohemial proesses in atfish milt not extended with ryopreservation materials. This was what proaly resulted in the poor milt quality reorded at 15 minutes of storage and the deteriorated motility oserved after 15 minutes of storage even with refrigeration. As a result, Fridge and Freeze temperatures may not e feasile in preserving or storing unproessed atfish milt. REFERENCES [1] Tengels G. G. A., systemati revision of the Afrian speies of the genus Clarias (Pises: Clariidae). Annals Musee. Revue. Afrian. Contriutions, 247, 199 (1986). Availale at (last aessed 26 August, 2012). [2] Sydenham D. H. J., New speies of Clarias from West Afria. Rev. Zool. Afr., Vol. XCIV, Fas., 31, (1980). [3] William R., Fish and Fisheries of Northern Nigeria. Ministry of Agriulture, Northern Nigeria. Gaskiya Corporation, Zaria. Pp. 83 (1967). [4] Heht T., Oellermann L. and Verheust L., Perspetives and Clariid atfish ulture in Afria. In: Legendre, M. & Proteau, and J-P. (Eds.). The Biology and Culture of Catfishes. Aquati Living Resoures. 9, (1996). Paris, Hors Series. International Journal of Innovative Studies in Aquati Biology and Fisheries (IJISABF) Page 10

5 Effet of Storage Temperature on Afrian Catfish (Clarias Gariepinus Burhell 1822) Milt Quality [5] De Graaf G. J., Galemoni F. and Banzoussi, B., The artifiial reprodution and fingerling prodution of the Afrian atfish Clarias gariepinus (Burhell 1822) in proteted and unproteted ponds. Aquaulture Researh. 26, (1995). [6] FAO. Handook on the artifiial reprodution and pond rearing of the Afrian atfish Clarias gariepinus in su-saharan Afria. Food and Agriultural Organization, Fisheries Tehnial Rome, Paper 362 (1996). [7] Idahor K. O., Yakuu A. Umar A. F. Yahaya F. and Musa. L. A., Morphometri traits, spermatozoa mirosopy and stripped eggs fertilization with milt olleted from live Afrian Mudfish (Clarias gariepinus). Livestok Researh for Rural Development. 26, (8) (2014). idah26150.htm. [8] Amupitan P. S., Ilori O. I., Aladele S. E., Odofin W. T. and Omitogun, O. G., Saling up the viaility of royo reserved giant atfish semen in liquid nitrogen for ommerial appliation. Proeedings of the 34th Annual Conferene of the Geneti Soiety of Nigeria. NIHORT, Iadan. Pp (2010). ISSN [9] Riley K. L. P., Refrigerated storage and ryopreservation of sperm for the prodution of Red snapper and Snapper hyrids. Master s Thesis, Agriulture and Mehanial College, Louisiana University. USA (2002). [10] Bruton M. N., Alternative life-history strategies of atfishes. In: Legendre, M. and J. P. Proteau (Eds.).The Biology and ulture of Catfishes. Aquati Living Resoures. Paris, Hors Series. 9, (1996). [11] Legendre M., Seasonal hanges in sexual maturity and feundity, and HCG-indued reeding of the atfish Heteroranhus longifilis Val. (Clariidae) reared in Erien lagoon (Ivory Coast). Aquaulture. 55, (1986). [12] Hogendoorn H., Controlled propagation of the Afrian atfish, Clarias lazera (C&V). I. Reprodutive iology and field experiments. Aquaulture. 17, (1979). [13] Idahor K. O. Mirosopi oservation of spermatozoa in milt olleted with syringe without sarifiing the male Afrian Catfish (Clarias anguillaris B. 1911). International Journal of Fisheries and Aquati Studies. 2, (2014). [14] Omitogun O. G., Oyeleye O. O., Betiku C. O. Ojiokpota C., Aladele S. E. and Sarumi M. B., Potentials of short-term ryopreserved sperm of the giant Afrian atfish, Clarias gariepinus (Burhell, 1822) for aquaulture in Nigeria. Pro. 31st Ann. Conf. of Geneti Soiety of Nigeria, Moor Plantation, Iadan. Pp (2006). [15] Viveen W. J. A. R, Rihter C. J. J, van Oordt, P. G. W. J, Jansen J. A. L. and Huisman, E. A., Pratial Manual for the Culture of the Afrian Catfish (C. gariepinus) 2500 EB, The Hague, The Netherland. Pp. 94 (1996). [16] Tonguthai K, China S, Limsuwan C, Somsiri T, Chanrathakool P, Kanhana khan et al. Handook of hyrid atfish: Husandry and health. Aquati Animal Health Researh Institute, Department of Fisheries, Kasetsart University Campus, Jatujak, Bangkok 10900, Thailand. Pp. 37 (1993). [17] Hogendoorn H., Controlled propagation of the Afrian atfish, Clarias lazera (C&V). III. Feeding and growth of fry, Aquaulture, 21, (1980). [18] Boe J. and Lae C., Egg and Sperm Quantity in Fish. General and Comparative Endorinology. 165, (2010). [19] Suquet M., Dreanno C., Fauvel C., Cosson J. and Billard R., Cryopreservation of sperm in marine fish. Aquaulture Researh, 31, (2000). [20] Uranyi B., Horvath A., Varga Z. and Horvath L., Effet of extenders on sperm ryopreservation of Afrian atfish, Clarias gariepinus (Burhell). Aquaulture Researh. 30, (1999). [21] Egunike G. N., A toast for spermatozoa: One half the story of mammalian life. An Inaugural Leture Series, University of Iadan, Iadan Nigeria. Pp. 58 (1995). [22] SIGMA, Cell ounting and ell viaility. Saint Louis, MO Pp (1994). USA. [23] Prathalingam N. S. Holt W. W., Revell S. G., Jones S. and Watson P. E., The preision and auray of six different methods to determine sperm onentration. J. Andrology. 27, (2006). DOI: /jandrol [24] SAS, Statistial Analysis System, User s Guide. Statistial. Version 9.1 e4d. SAS. Inst. In. Cary. N.C. USA (2012). [25] FAO, Preservation tehniques: Fish utilization and trade. Fisheries and Aquaulture Department, Rome. Food and Agriulture Organization of the United Nations for a world without hunger. (2017). Availale at (last aessed 21 Novemer 2017). [26] Okunseor S. A and Idahor K. O., Use of morphologial and haematologial indies of Afrian Mudfish (Clarias gariepinus) to assess Doma Dam water quality. Proeedings of the 14th Annual Conferene of Animal Siene Assoiation of Nigeria, LAUTECH, Ogomoso. Pp (2009). International Journal of Innovative Studies in Aquati Biology and Fisheries (IJISABF) Page 11

6 Effet of Storage Temperature on Afrian Catfish (Clarias Gariepinus Burhell 1822) Milt Quality [27] Idahor K. O., Anatomial study of testes position in Afrian Mudfish (Clarias gariepinus): guide to milt harvesting without sarifiing the fish. Journal of Animal Prodution Advanes. 3, (2013). DOI: /japa [28] Ayoola S. O., Relationships of hemial omposition, quantity of milt to fertility and hathaility of Clarias gariepinus (Burhell, 1822). Afrian Journal of Food, Agriulture, Nutrition and Development. 9, (2009). [29] Bart A. N. and Dunham R. A. Effets of sperm onentration and egg numer on fertilization effiieny with hannel atfish (Italurus puntatus) eggs and lue atfish (I. furatus) spermatozoa. Theriogenology. 45, (1996). [30] Muhlisin Z. A., Hashim R. and Chong, A. S. C. Preliminary study on the ryopreservation of tropial agrid atfish (Mystus nemurus) spermatozoa; the effet of extender and ryoprotetant on the motility after short-term storage. Theriogenology. 62: (2004). Citation: K. O. Idahor, et al., " Effet of Storage Temperature on Afrian Catfish (Clarias Gariepinus Burhell 1822) Milt Quality", International Journal of Innovative Studies in Aquati Biology an d Fisheries, vol. 4, no. 1, p. 7-12, Copyright: 2018 Authors. This is an open-aess artile distriuted under the terms of the Creative Commons Attriution Liense, whih permits unrestrited use, distriution, and reprodution in any medium, provided the original author and soure are redited. International Journal of Innovative Studies in Aquati Biology and Fisheries (IJISABF) Page 12

Spermatozoa Revival with Addition of Diluted Dextrose Saline in African Catfish (Clarias gariepinus Burchell 1822)

International Journal of Research Studies in Zoology Volume 4, Issue 2, 2018, PP 7-11 ISSN No. 2454-941X DOI: http://dx.doi.org/10.20431/2454-941x.0402002 www.arcjournals.org Spermatozoa Revival with Addition