AN EVALUATION OF THE OUT COME OF BULL CASTRATION BY INTRA-TESTICULAR INJECTİON OF ETHANOL AND CALCIUM CHLORIDE - PDF

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AN EVALUATION OF THE OUT COME OF BULL CASTRATION BY INTRA-TESTICULAR INJECTİON OF ETHANOL AND CALCIUM CHLORIDE Canpolat İ. * Bulut S.* Günay C. * Eröksüz H.** Gür S.*** * Department of Surgery, Veterinary

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AN EVALUATION OF THE OUT COME OF BULL CASTRATION BY INTRA-TESTICULAR INJECTİON OF ETHANOL AND CALCIUM CHLORIDE Canpolat İ. * Bulut S.* Günay C. * Eröksüz H.** Gür S.*** * Department of Surgery, Veterinary Faculty, Firat University, Elazig- Turkey ** Department of Pathology, Veterinary Faculty, Firat University, Elazig- Turkey *** Department of Reproduction and artificial insemination, Veterinary Faculty, Firat University, Elazig- Turkey SUMMARY This study evaluated intra-testicular injection of ethanol and calcium chloride for chemical-castration in 12 mix bred young bulls ( kg). Ten ml absolute ethanol and ten ml calcium chloride at 30% concentration per testis were injected intratesticularly and the testicles were removed with the open surgical technique about 60 days for histopathologic evaluation. Testicular swelling was evident in both group bulls following injection and reached peak within 48 hours. While testicular volume decreased significantly (P 0.05) in ethanol group after 3 weeks, no significant change occurred in calcium chloride group. The testicles underwent atrophy at the 60 th day in ethanol group with no marked alteration in calcium chloride group. Though only 3 bulls were sterile, other bulls maintained androgenesis. It was concluded that intra-testicular injections of ethanol and calcium chloride administration may not be accepted as a suitable alternative to the open surgical technique for castration in bulls. INTRODUCTION Chemosterilization is tried in males monkeys, hamsters, rabbits, rats and dogs by intratesticular injection of some agents such as ferric chloride (Kar et al.,1965), danazol (Dixit et al., 1975), BCG (Das et al.,1982), zinc tannate (Fahim et al.,1982), glycerol (Weinbauer et al., 1985, Immegart 2000), glucose, NaCl (Heath et al., 1987, Russell 1987 et all.) DBCP (Shemi et al.,1988), lactic acid (Fordyce et al.,1989), zinc arginine (Fahim et al.,1993), sodium fluoride (Sprando et al.,1996), formaline (Bakır et al.,2002) and calcium chloride (Samanta 1998, Jana et al. 2002), potassium permanganate+glacial acetic (Giri et all.,2002). In male ruminants intratesticularly lactic acid (Hill et all. 1985) tannic acid and zinc sulphate (Feher et all. 1985) alpha-hydroxypropionic acid (Cohen et all. 1995), Formalin (Ijaz et all. 2000) Castrate-Quin 14 (Soerensen et all. 2001) have been tried, but because of many complications following the use of these chemicals, an effective chemosterilizing agent has yet to be established. The purpose of the study was to determine the efficacy of intratesticular injection of ethanol and calcium chloride on chemosterilization outcome of the bulls. MATERIALS AND METHODS In this study, 12 mixbreed young bulls ( kg) were used. They were divided randomly and equally into two groups called EG (ethanol group) and CCG (calcium chloride group). Ten ml absolute ethanol and ten ml calcium chloride at 30% concentration per testis were injected intra-testicularly. Trans-scrotal testicular ultrasonographv was performed with Mhz linear probe (Pie medical-scanvet 200) prior to the intratesticular injection and once a day for week following intratesticular injection and then one week intervals up to the end of the study. During this examination, the ultrasonographic appearance of a testis was evaluated and its height (h), width (W) and length (l) was measured. The volume of the testis was estimated using Volume= (π/6) X (w X h X l) equation. Sement examination and serum testosterone concentrations was performed prior to and 15, 30 and 60 days after intratesticular drug injection. The testicles were then removed the open surgical technique about 60 days after drug injection for histopathologic evaluation. Statistical analyses was performed with student t-test. RESULT Testicular swelling was evident in both group bulls following injection and reached peak within 48 hours. While testicular volume decreased significantly (P 0.05) in EG after 3 weeks, no significant change occurred in CCG (P 0.05). The testicles underwent atrophy at the 60 th day in EG but marked alteration was not observed in CCG (Figure 1). In 2 CCG cases, testicular swelling was associated with orchitis and scrotal sloughing. At necropsy, their testicular tissues were found to had necrotized and abcess formation (Fig-2). These problems were not recognized in any EG cases. Ultrasonographic examination following drug injection by revealed that the testicular tissue of both group animals had a diffuse echotexture and increased echogenite (Fig-3). The necrotized regions were differentiated ultrasonographically from the normal testicular tissue with a hypoechogenic area (Fig-4) In this study, mean baseline value of serum testosterone concentration was determined as 13.2 ± 2.6 pg/ml in EG and 13.0 ± 2.4 CCG which reduced significantly P 0.05 in EG and remained almost the same in CCG (P 0.05). Intratesticular injection of etanol produced a more gradual fall: from baseline 13.2 ± 2.6 pg/ml to 5.7 ± 3.9, 3.5 ± 0.7 and 2.6 ± 0.7 pg/ml 15, 30 and 60 days, respectively. On the other hand calcium chloride remained almost the same : 12.9± 1.8, 13.0 ± 2.2 and 12.9± 2.4 pg/ml 15, 30 and 60 days, respectively. In the sement examination only 3 case had inactive sperm in EG, the remaining case had active sperm.. In the microscobic examination of EG, severe diffuse tubular necrosis along with varying degree of inflammatory response was the main finding observed (Fig-5). Inflamatory cells were consisted of mainly mononuclear cells. Intertubular edema, fibrosis, hemorrhage were also detected. Some of the necrotic cells showed desquamation, or even calcification. Intertubuler vessels were severely congested. Although similar lesions were noted in CCG bulls, the severity and distribution of the lesions were not so pronounced as in EG group. DISCUSSION Chemical, surgical, and mechanical castration, were tried in bulls for chemical castration in male ruminants intratesticular lactic acid (Hill et all. 1985) tannic acid and zinc sulphate (Feher et all. 1985) alpha-hydroxypropionic acid (Cohen et all. 1995), Formalin (Ijaz et all. 2000) Castrate-Quin 14 (Soerensen et all. 2001) have been used. A few studies on the clinical use of absolute ethanol have been reported. These studies involved in the treatment of simple renal cysts, tumors and renal angioinfarction (Ellman et al., 1981 Livraghi et al., 1986) and benign prostate hyperplasie (Zvara et al., (1999). The actual physiologic effect of this treatment has previously been speculated. Necrosis and infarction leading to fibrosis, shrinkage, or sloughing of tissue are presumed to be its mechanism of action (Zvara et al.,1999). In this study; severe diffuse tubular necrosis along with varying degree of inflammatory response was the main finding observed (Fig 5). For chemical sterilization ethanol and formaldehyde mixture is also used (Plant et all. 1979, Gardner 1980). Gardner (1980) used inratesticullarly a solution consisting of 3.6% formaldehyde in 90% ethanol for sterilization purpose in 10 bulls where 3 showed active sperms after 85 days. In the present study, absolute ethanole was used in 6 bulls while 3 became sterilized others maintained androgenesis despite marked testicular atrophy. Optimum effective doses of calcium chloride in rat were considered to be between mg, however, the most desired result obtained with 20 mg (Jana et al.,2002). These authors and Samanta (1998) found marked necrosis in the seminiferous tubules during the use of this agent. In a study; the effect of calcium chloride on serum testosterone levels following intratesticular administration was investigated (Mitra and Samanta 2001) and found a marked reduction in serum testosterone level in bulls. The result of the present study, however, appear to disagree with the former one. Because, the alteration in serum testosterone level in CCG was minimum and histopathological disorders were not serious. Thus, this agent has in fact no effect on the chemosterilisation of the bulls. It was concluded that intra-testicular injections of ethanol and calcium chloride administration may not be accepted as a suitable alternative to the open surgical technique for castration of bulls. Fig 1: Left CCG and right testicles underwent atrophy at the 60 th day in EG. Fig 2: The testicular tissues had necrotized and abcess formation in CCG. Fig 3: The all animals of both groups to present diffuse echotexture and increased echogenite after intratesticular injections. in EG. Fig 4: The testicular tissue with hypoechogenic appearance in the necrotic area (HEX20) Fig 5: Diffus tubular necrosis and post necrotic mononuclear cell infiltration. REFERENCES Bakır B,Gülyüz F,Karaca F,Yüksel H,Şahin A,Uslu BA: Chemical castration in dogs. YYU J Healty Sciences. 8(1-2)6-9,2002 Cohen-RDH; King-BD; Thomas-LR; Janzen-ED Efficacy and stress of chemical versus surgical castration of cattle. Canadian-Journal-of-Animal-Science Das RP, Mustafa AS, Talwar GP Atrophy of seminiferous tubules of mouse testes after intratesticular injection of BCG and their regeneration.arch Androl 1982 Nov;9(3): Dixit VP, Lohiya NK, Arya M, Agrawal M.Chemical sterilization of male dogs after a single intra-testicular injection of Danazol .Folia Biol (Krakow). 1975;23(3):305-10 Ellman B.A., Parkhill B.J., Curry T.S. et al., (1981) Ablation of renal tumors with absolute ethanol: a new technique. Radiology 141, 619 Fahim MS, Fahim Z, Harman JM.Chemical sterilization in the male part I: rats. Arch Androl 1982 Nov;9(3):261-5 Fahim MS, Wang M, Sutcu MF, Fahim Z, Youngquist RS.Sterilization of dogs with intra-epididymal injection of zinc arginine. Contraception 1993 Jan;47(1): Feher-T; Bodrogi-L; Makray-S Dynamics of serum testosterone levels in bulls: daily, seasonally and after chemical or surgical castration. Magyar-Allatorvosok-Lapja. 1985, 40: 1, Fordyce G, Hodge PB, Beaman NJ, Laing AR, Campero C, Shepherd RK.An evaluation of calf castration by intra-testicular injection of a lactic acid solution. Aust Vet J 1989 Sep;66(9):272-6 Gardner IA, Sclerosing sterilisation in bulls. New-South-Wales-Veterinary-Proceedings. 1980, 16: Giri-SC; Yadav-BPS; Panda-SK Chemical castration in pigs. Indian-Journal-of-Animal- Sciences. 2002, 72: 6, Heath E, Arowolo R.The early histopathologic effects of intratesticular injection with hyperosmolar glycerol, glucose or NaCl solutions. Andrologia 1987 Nov- Dec;19(6): Hill-GM; Neville-WE Jr.; Richardson-KL; Utley-PR; Stewart-RL; Castration method and progesterone-estradiol implant effects on growth rate of suckling calves. Journal-of- Dairy-Science. 1985, 68: 11, Immegart HM, Threlfall WR.Evaluation of intratesticular injection of glycerol for nonsurgical sterilization of dogs. Am J Vet Res 2000 May;61(5):544-9 Jana K, Samanta PK, Ghosh D.Vet Res Commun 2002 Dec;26(8):651-73Dosedependent response to an intratesticular injection of calcium chloride for induction of chemosterilization in adult albino rats. Kar AB, Kamboj VP and Goswami A (1965): J Rep and Fert 9,115-7 Livraghi, T, Festi, D. Monti F. US-guided percutaneous alcohol injection of small hepatic and abdominal tumors. Radiology 161 (1986), pp Ijaz-A; Abalkhail-AA; Khamas-WAH Effect of intra testicular injection of formalin on seminiferous tubules in Awassi lambs. Pakistan-Veterinary-Journal. 2000, 20: 3, Mitra-B; Samanta-PK Changes in testosterone level in scrub bulls by intratesticular administration of calcium chloride. Indian-Journal-of-Veterinary-Medicine. 2001, 21: 2, 98 Plant JW, Seaman JT, Jakovljevic D.Non-surgical sterilisation of rams using a sclerosing agent. Aust Vet J 1979 Jun; 55 (6) : Russell LD, Saxena NK, Weber JE.Intratesticular injection as a method to assess the potential toxicity of various agents and to study mechanisms of normal spermatogenesis. Samanta PK Chemosterilization of stray dogs. Indian-Journal-of-Animal-Health. 1998, 37: 1, 61-62; 4 Shemi D, Marx Z, Kaplanski J, Potashnik G, Sod-Moriah UA.Testicular damage development in rats injected with dibromochloropropane (DBCP). Andrologia 1988 Jul- Aug;20(4):331-7 Soerensen-B; Soutello-RVG-de; Gomes-CR-de-A; Garcia-R; Marini-A; Fonzar-JF; Paes-F; Rasteiro-VS; Souza-Filho-WF-de; Tozi-CR; Gasparelli-Junior-AG; Correa-BG; Moitinho-MP; de-a-gomes-cr; de-soutello-rvg; de-souza-filho-wfchemical, surgical and mechanical castration of bovine. Study comparative. Ciencias-Agrarias-eda-Saude. 2001, 1: 2, 7-10; 6 Sprando RL, Black TN, Ames MJ, Rorie JI, Collins TF.Effect of intratesticular injection of sodium fluoride on spermatogenesis..food Chem Toxicol 1996 Apr;34(4): Weinbauer GF, Galhotra MM, Nieschlag E.Focal testicular destruction following intratesticular injection of glycerol in rats. Int J Androl 1985 Oct;8(5): Zvara, P., Karpman, E., Stoppacher, R. et al: Ablation of canine prostate using transurethral intraprostatic absolute ethanol injection. Urology, 54: 411,1999
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