EFFECT OF SOME ANTIBIOTICS ON REPRODUCTIVE PERFORMANCE, BLOOD CONSTITUENTS AND ORGANS-HISTOPATHOLOGY OF BAUSCAT BUCK RABBITS.

 

S. A. Gad Alla¹; Y.E. El-Bolkiny²; Merveet A. Mansor²  and Nubile  E.  El-Kasas¹

¹Animal Production Research Institute, Agricultural Research Center, Gaza, Egypt.

²Department of  Zoology, Facility of Science, Tanta University, Tanta, Egypt.

 

ABSTRACT

 Thirty six Basket buck rabbits, 6 months of age  with an average initial body weight, 2835 ± 44.58 g were randomly allotted into three experimental groups (N= 12) based on body weight. The 1^st group was used as control and injected with saline solution (1 ml of 0.09 % NaCL). The 2^nd and 3^rd groups were injected subcutaneous with 2.5 mg of each tetracycline and enrofloxacin per kg live body weight biweekly for 10 weeks, respectively. Semen samples were individually collected from the five bucks once weekly for a period of 10 weeks to evaluate semen quality. Blood samples from five Basket buck rabbits in each experimental group were collected biweekly throughout the experimental period, also blood parameters were studied. At the end of experiment, three bucks from each experimental groups were randomly chosen and slaughtered to study the histopathological examination. Specimens of liver, kidneys and testes were taken from each animal for the histopathological investigation.

   The present results showed that the live body weight and the relative weight of liver, testes and epididymis was not affected significantly by injection with tetracycline or enrofloxacine. However, the relative weight of kidney of bucks which treated with tetracycline and enrofloxacine increased significantly       (P < 0.05) as compared with the control group. The treatment of bucks with  tetracycline or enrofloxacine decreased insignificantly the total number of RBCs and WBCs as compared with the control group. The total number of platelets were decreased (P < 0.05) in rabbits treated with tetracycline and increased  (P< 0.05) in which treated with enrofloxacine. After treatment with tetracycline and enrofloxacine, the mean concentration of hemoglobin (Hb) and packed cell volume (PCV) were similarly with control.

  The total protein, globuline, bilirubin levels and createnin were decreased (P < 0.05) with rabbits treated with tetracycline or enrofloxacine compared to the control group. However, albumin, albumin / globuline (A/G) ratio and the plasma  urea –N concentrations were similar in both treated with tetracycline or enrofloxacine and control groups.

Injection of bucks with tetracycline induced (P < 0.05) reduction of plasma AST activity and increased (P < 0.05) of plasma ALT activity. However, injection of rabbits with enrofloxacine decreased (P < 0.05) of plasma AST and ALT activity as compared with the control group.The testesterone hormone  was increased significantly after injection rabbits with tetracycline or enrofloxacine groups as compared with the control group.

Gross and microscopic examination of kidney, liver and testicular of Bauscat buck rabbits showed that injection with tetracycline or enrofloxacin had no adverse effect on these organs, which appeared histopathologically similar to those of the control rabbits. All physical semen characteristics improved significantly (P<0.01) for treated rabbits with tetracycline or enrofloxacine compared to the control group. However, the ejaculate volume were not affected significantly  by treated with both drugs.

     Key words: Tetracycline, enrofloxacin, and physical semen traits, blood components, organs-histopathology.

 

INTRODUCTION

Rabbits can be driven to health problems  and mortality due to an imbalance  of the digestive flora, under the influence of environmental factors. In order to prevent this health problems several products are available on the market and some are employed more or less extensively in the practice.

Antibiotic is a group of chemical compounds produced biologically by certain microorganisms which posses bactericidal effects. Antibiotic are added to feed of several classes of livestock to prevent or control diseases, to stimulate growth and improve feed efficiency (Miles et al., 1984). Antibiotic have been used widely as growth promoters particularly of antimicrobial properties (El-Sherbiny et al., 1990; Dorgham et al., 1994 and Radwan et al., 1996). Among the wide range of antibiotics which has been tested tetracycline or enrofloxacine used on a preventive or on a curative basis.

Tetracycline known as the broad spectrum antibiotics because have their affect on gram positive and negative bacteria, as well as,  a few non bacterial species. It had protective effects against chronic aflatoxicosis enhanced by cysteine and methionine in rabbits (Clark et al., 1982). Also, due to chelating properties of tetracycline it has  been used as in vivo indicator of new bone formation because it forms complexes with mineral at bone forming surfaces (Moalli et al., 1994).

Enrofloxacine is abroad spectrum from fluoroquinolone antibiotics. It is routinely used to treat bacterial infections (Verma et al., 1999 and Oluch et al.,2001). It has the highest efficacy against canine E coli isolates from dogs. Also, Lautzenhiser et al. (2001) studied the effect of enrofloxacin and ciprofloxacin against E coli and staphylococcal isolated from dogs and the in vivo transformation of enrofloxacin to ciprofloxacin may enhance the efficacy of enrofloxacin.

Therefore, the aimed of the present study to investigate the effect of injection  of  some antibiotics (tetracycline and enrofloxacin) subcutanously on reproductive performance, some blood constituents and organs- histobathology of Bauscat buck rabbits.

 

MATERIALS AND METHODS

 

The experimental and laboratory works of the present study were carried out at El-Gemeza Experimental Station, Animal Production, Research Institute,Ministry of Agriculture,Egypt,  at March, 2002 and extended  for 10 weeks.

The present study lasted 10 weeks in order to evaluate the effects of injection of some antibiotics (tetracycline and enrofloxacin) subcutanously on reproductive performance, some blood constituents and organs- histopathology of Basket male rabbits. For this purpose, thirty six Bauscat buck rabbits, 6 months of age with an average initial body weight, 2835 ± 44.58 g were randomly allotted into three experimental groups (N= 12) based on body weight. The 1^st group was used as control and injected with saline solution (1 ml of 0.09 % NaCL). The 2^nd and 3^rd groups were injected subcutanous with 2.5 mg of each tetracycline ( 1 ml of an injectable solution contained 2mg tetracycline that manfactured by Pfizer Co. Cairo, Egypt) and enrofloxacin (1 ml of an injectable solution contained 2mg enrofloxacin that manfactured by Alexandria Co. for Pharmaceuticals, Alexandria, Egypt.)  per kg live body weight biweekly for 10 weeks, respectively.

The rabbits were housed individually in wire cages (60 x 55 x 40 cm) provided with galvanized feeders and nipple drinkers. Fresh drinking water were avaliable all time. The animals were kept under the same environmental and managerial conditions. The basal ration was formulated in one of feed mills to meet the nutrient requirements of rabbits according to NRC (1977). The ration was offered to rabbits ad libitum. The ingreadients and chemical composition of the pelleted ration are shown in Table (1).

Five Bauscat buck rabbits were randomly chosen from each experimental group. Semen samples were individually collected from the bucks once weekly for a period of 10 weeks by using an artificial vagina. Semen quality of bucks was determined including ejaculate volume (ml), wave motion (scores), sperm motility (%), sperm concentration (x 10⁶/ml), dead and abnormal spermatozoa (%) as described by El-Gaafary (1987) and El-Kelawy (1993).

Blood samples from five bucks in each experimental group were collected biweekly  throughout the experimental period. At each collection 5 ml of blood were drained and divided into two tubes. The 1^st tube was without heparine and the 2^nd sample was placed into heprinized tube.

Blood samples were without heparin were immediately centrifugation at 3000  r.p.m.  for 15 minutes  and  stored  frozen  (-20⁰C)  in plastic vials until  performance of  the  biochemical tests. Levels  of   serum total protein and  albumen  was determined  according to Henery (1964) using commercial kits obtained from Diamond Diagnostics. The globulin values were obtained by substracting the values of albumin from the total protein and the values of A/G ratio was also calculated. Urea-N, creatinine and total bilirubin concentration was determined by using commercial Kits of Diamond Diagnostics according to the method of Henery (1974). The activity of aspartate aminotransferase (AST) and alanine  aminotrnsferase (ALT)  was assayed  according to the method described by Reitman and Frankel (1957). The levels of serum blood testesterone was also determined by radioimmiunoassay technique, according to the method  described by Jaffe and Behrman (1974) and Wilson et al. (1992) using coated kits purchased from Diagnostic Products Corporation Kits, Los Angles, U.S.A.

At the end of experiment, three bucks from each experimental groups were randomly chosen and slaughtered to study the histopathological examination. Specimens of liver, kidneys and testes were taken from each animal for the histopathological investigation. The specimens were fixed in a 10 % formalin saline, embedded in paraffin wax and sectioned at 5 microns thickness sections which stained with hematoxylin and eosin (Carleton, 1967).

Data of the experiment were statistically analyzed according to Snedcor and Chochran (1982), using SPSS (1998). The differences between means were tested using Duncan^,s New  Multiple  Rang Test  (Duncan,1955).   

 

Tble  (1): Ingreadients and chemical composition of the pelleted rations.

* Each kg of  vitamin and mineral provides: 4.000.000 Vit A, 5000.000 IU Vit D,16.7g Vit E, 0.67g Vit K, 0.67g Vit B₁, 2.0 g Vit B₂, 0.67g B₆, 0.004g Vit B₁₂, 16.7g Niacin, 6.67 Pantothinic acid, 0.07g Biotine, 1.67 g Folic acid, 400 g Choline chloride, 23.3g Zn, 10g Mn, 25 g Fe, 1.67g Cu, 0.25g I, 0.033g Se and 1.33 g Mg.

RESULTS AND DISCUSSION

Effect of tetracycline and enrofloxacine on:

Body weight and relaive weight of organs:

            Data in Table 2 showed that the live body weight was not affected significantly (P< 0.05) by injection with tetracycline or enrofloxacine. Moreover, the relative weight of liver, testes and epididymis showed insignificant differences between treated groups with tetracycline or enrofloxacine and untreated group (control). However, the relative weight of kidney of Bauscat buck rabbits which treated with tetracycline and enrofloxacine increased significantly (P < 0.05) as compared with the corresponding value of the untreated group.Similar results were reported by Abdel-Lateef et al. (1998) and El-Banna and Ibrahim (1999) on body weight and Izat et al. (1989) on the relative weight of investigated organs (Liver, kidney and testes).

 

 

 

 

 

 

Table 2. Effect of tertracycline and enrofloxacin  on body weight and relative weight of some organs (X±SE)  of Bauscat buck  rabbits      

Means in the same row having different letters are significantly differ, (P<0.05).

* = P< 0.05,            N.S= Not significant.

 

 

Effect of tetracycline and enrofloxacine on the blood count:

The results in Table 3 showed that the treatment of Bauscat buck rabbits with 2.5 mg tetracycline or enrofloxacine decreased insignificantly  the total number of RBCs as compared with the control group. Also, WBCs was not affected significantly by the treatment with both drugs. On the other hand, the total number of platelets were decreased significantly (P < 0.05) in Bauscat buck rabbits treated with 2.5 mg tetracycline and increased (P< 0.05) in which treated with 2.5 mg enrofloxacine. Similar results were obtained by Flescher et al. (2000) found that chickens fed diet   supplemented with enrofloxacin had no changes in RBCs and WBCs. However, El-Banna and Ibrahim (1999) showed that growing Bauscat rabbits fed on diet supplemented with similar antibtics increased significantly RBCs count. However, Dniyog and Bhowmik (2003) found that broiler chickens fed with ciprofloxacin decreased significantly RBCs count at 7 days post treatment, but it had no significant effect on WBCs count indicating anaemia, which possibly resulted from either its toxic depression on bone narrow or its suppressive effect on hemamopoietic tissue.

Table 3. Effect of tertracycline and enrofloxacin  on blood count and omponents (X±SE)of male Bauscat  rabbits.

Means in the same row having different letters are significantly differ, (P<0.05).

* = P< 0.05,  N.S= Not significant.

 

After treatment with tetracycline and enrofloxacine, the mean concentration of hemoglobin (Hb) and packed cell volume (PCV) were similarly with untreated group (control). Flescher et al. (2000) reported that chickens fed a diet supplemented with enrofloxacin had a significant (P < 0.01) increase in HB, but no significant effect on PCV. On the other hand, Dniyog and Bhowmik (2003) found that broiler chickens that fed with ciprofloxacin led to a significant decrease in Hb and PCV at 7 and 14 days post treatment. However, El-Banna and Ibrahim (1999) showed that the feeding growing rabbits with such antibiotics increased significantly the Hb and PCV values.

 

Serum biological synthetic activity:

The present results in Table 3 showed that the total protein, globuline and bilirubin levels were decreased significantly (P < 0.05) with Bauscat rabbits treated with tetracycline or enrofloxacine compared to the corresponding values of the control group. However, albumin and albumin/globuline (A/G) ratio were not affected significantly by treatment with both drugs. El-Tantawy et al. (2001) found that total protein, albumin and globulin levels were increased significantly in Bauscat rabbits  supplemented with flavomycine than the corresponding values of the untreated group (control). El-Yamany et al. (1999) found that oxytetracycline led no effect on total protein, albumin, globulin and A/G ratio of one-day old ducks. Oppositly, Dniyog and Bhowmik (2003) found that plasma total protein and A/G ratio were significantly decreased in chickens treated with ciprofloxacin.

The present results showed that either tetracycline or enrofloxacine tend to decrease the proteins fractions and it was possible because of its decreased synthesis by degenerated and/or necrotic liver resulting in low protein level in blood. It is possible because of decreased in glucose level by gluconeogenesis and this leads to loss of plasma proteins. On the other hand, both drugs increased the plasma level of the protein fractions and this result could be interpreted on this basis of little hepatocyte degeneration, which led to leakage of protein into circulation.

 

Kidney functions:

The present data showed that the plasma urea –N concentrations were similar in both traated bucks with tetracycline or enrofloxacine and untreated group (control). However, creatinin concentrations decreased significantly (P < 0.05) of treated bucks with both drugs as compared with the corresponding values of the control group (Table 3). Similar results were reported by El-Koly and El-Hady (1996) reported that treatment with flavomycine had no effect on plasma urea-N  and createnine concentrations of buffaloes and Friesian calves. However, El-Amary et al. (2001) showed that the effect of avoparcin increased the plasma urea-N concentration after feeding of sheep with 0.75 mg/day. Also, El-Banna and Ibrahim (1999) found that urea-N and createnine levels of growing Bauscat rabbits supplemented with verginiamycine were decreased significantly.  

Gross and microscopic examination of kidney of bucks showed that  injection with tetracycline or enrofloxacin had no adverse effect on the kidney, which appered histopathologically similar to those of the control rabbits.Tissue examination with a light microscopy revealed that there was no any change in the kidney section of rabbits treated with either tetracycline or enrofloxacin (Figure 1). The microscopic examination of kidney of male Bauscat rabbits reflected a normal appearance without pathological observations. The renal corpuscle consists of a well developed glomerulus and a prominent Bowman^,s capsule. Also, the renal tubules appeared with normal epithelia in both the cortical and outer meullar regions, except some lymphocitic infiltration and mild tubular degeneration.

 

Liver functions:

Injection of Basket buck rabbits with tetracycline  induced a significantly        (P<0.05) reduction of plasma AST activity and increased significantly (P < 0.05) of plasma ALT activity. However, injection of bucks with enrofloxacine decreased significantly  (P < 0.05)  of  plasma   AST   and   ALT   activity   as  compared  with  the

 

 

A

 

 

 

 

 

 

 

 

 

B

 

 

 

 

 

 

 

 

 

 

C

 

 

 

 

 

 

 

 

Figure 1. Cross section in kideny of bucks injected with tetracycline (A) and enrofloxacine (B) as compared with untreated  (C).

 

 

 

 

 

corresponding values of the control group (Table 3). Dniyog and Bhowmik (2003) showed that ciprofloxacin had  corresponding values of the control group (Table 3). Dniyog and Bhowmik (2003) showed that ciprofloxacin had significant increase of surum AST and ALT for broiler chickens. El-Tantawy et al. (2001) found that serum AST and ALT of bucks were not affected significantly by traetment with flavomycine.

            Gross and microscopic examination of liver of bucks showed that  injection with tetracycline or enrofloxacin had no adverse effect on the liver, which appered histopathologically similar to those of the control rabbits. Microscopic examination revealed that the liver contains normal hepatic lobules with normal hepatic cell atands enclosing a central vein and normal appearance (Figure 2). Moreover, hepatocytes appeared with more intense esinophilic cytoplasm and plastic regenerative in some slide and mild degeneration. These findings were in agreement with the results of the blood serum analysis (Table 3), which revealed normal liver functions of rabbits injected with tetracycline or enrofloxacin compared to the control.

 

Serum teasterone levels:

The present data showed that testesterone hormone  was increased significantly after injection with tetracycline or enrofloxacine in bucks as compared with the control group (Table 3). The lack effect of ciprofloxacin on circulating testesterone concentration may indicate that ciprofloxacin is a relatively selective inhibitor of mixed-function oxidase activity, affecting the activities of enzymes involved in the metabolism of substrates such as theophylline and antipyrine, but not the activities of those involved in testesterone synthesis. Ciprofloxacin may inhibit both the synthesis and degeneration of testesterone, resulting in little or no effect of circulating testesterone concentration. An analysis of testesterone precursor and metabolite concentration profiles would determine where this is a viable explantation. It  also possible that poor penteration of ciprofloxacin into the testes, the primary site of testesterone production, accounts for lack of effect on testesterone concentration.

 

Testicular histpathology:

Gross and microscopic examination of testicular of bucks showed that  injection with 2.5 mg tetracycline or enrofloxacin had no adverse effect on the testes, which appered histopathologically similar to those of the control rabbits. Microscopic examination revealed that there were no changes in the testis section of rabbit treated with either tetracycline or enrofloxacin (Figure 3). The normal testis which contains the wall of  seminiferous tubules in well developed and consistis of different layers of normal spermatocytes at different stages of development. The nuclei of sertoil cell often appear mid-way between the basement membranes of the seminiferous tubules and their lumena. The interstitial cells are appeared normal and seem to be surrounded by testicular lymph.

 

 

 

 

A

 

 

 

 

 

 

 

B

 

 

 

 

 

 

 

C

 

 

 

 

 

 

Figure 2. Cross section in liver of bucks injected with tetracycline (A) and enrofloxacine (B) as compared with untreated  (C).

 

 

 

 

A

 

 

 

 

 

 

B

 

 

 

 

 

 

 

C

 

 

 

 

 

 

 

Figure 3. Cross section in testes of bucks injected with tetracycline (A) and enrofloxacine (B) as compared with untreated  (C).

Physical semen characteristics:

Adverse effect of antibiotic agents on spermatogenesis or sperm functions have been demonestrated in animals and man (Dukes and Elis, 1982 ). In the present study all physical semen characteristics were improved significantly (P < 0.01) for treated bucks with tetracycline or enrofloxacine compared to the control group (Table 4). However, the ejaculate volume were not affected significantly by treated with both drugs. The obtained resulted cleared that treatment with either tetracycline or enrofloxacine tend to enhance the the sperme motility, and total sperm output. On the contarary, these drugs significantly decreased the percentage of dead and abnormal spermatozoa. In similar  study, Abbitt et al. (1984), Barth and Wood (1998) and Verma et al. (1999) cocluded that the ejaculate volume of semen did not change in beef bulls injected with either tetracycline or enrofloxacine.Abdel-Malak et al. (1999) found that oxytetracycline injection to Barki rams had no any significant changes in the percentage of live spermatozoa, which proved that oxytetracycline in therapeutic dose did not evoke any toxic effect on ram spermatozoa (Fattouh et al., 1991).

 

Table 4. Effect of tertracycline and enrofloxacin  on physical semen characteristics (X±SE)  of Basket buck  rabbits      

Means in the same row having different letters are significantly differ, (P<0.05).

** = P <0.01, * = P< 0.05,  N.S= Not significant.

 

In conclusion, the present results showed that injection of either of tetracycline or enrofloxacin with dose of 2.5 mg dose not change physiological and semen quality as growth promoters. Therefore, it can  be recommended that administration in low dose but prexautions should be taken to avoid over use, under environmental Egyptian conditions..

 

 

 

 

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تأثیر بعض التتراسیکلین و الانروفلوکساسین على معدل الأداء التناسلی ومکونات الدم والتغیرات الهستولوجیة فی ذکور الأرانب البوسکات

 

صلاح عبد الحکیم جاد الله¹ , یسرى السید البلقینی², میرفت أنور منصور² , نبیلة السید القصاص¹

1-  معهد بحوث الإنتاج الحیوانی – مرکز البحوث الزراعیة – وزارة الزراعة- ج.م.ع.

قسم علم الحیوان – کلیة العلوم – جامعة طنطا- ج.م.ع.

 

 

استخدم فی هذه الدراسة 36 أرنب ذکر بوسکات عمر 6 شهور متوسط الوزن 28350±44,58 جرام وزعت عشوائی فی ثلاث مجموعات تجریبی (12 أرنب/ مجموعة). المجموعة الأولى استخدمت کمجموعة مقارنة (کنترول) حققت بمعدل 1 مل من محلول ملحی فسیولوجى 0.09 %  کلورید صودیوم. المجموعة الثانیة و الثالثة حقنت تحت الجلد بمعدل 2.5 ملیجرام تحت الجلد / کجم  وزن حی من عقاری التتراسیکلین والانروفلوکساسین. جمعت عینات السائل المنوی أسبوعیا خلال فترة التجربة لدراسة جودة السائل المنوی . وأخذت عینات الدم من خمسة ذکور من کل مجموعة تجریبیة کل أسبوعین خلال فترة الدراسة وتم قیاس صفات الدم المختلفة. فی نهایة التجربة کما تم ذبح ثلاث ذکور عشوائیا من کل مجموعة تجریبیة لدراسة التغیرات الهستولوجیة للکبد والکلى والخصیتین.

أوضحت النتائج المتحصل علیها أن:

لم یتأثر وزن الجسم والوزن النسبی لکل من الکبد والخصیتین والبر بخ معنویا بالحقن التتراسیکلین والانروفلوکساسین بینما زاد الوزن النسبی للکلی معنویا فی الأرانب المعاملة بالتتراسیکلین والانروفلوکساسین مقارنة بمجموعة الکنترول. أدى الحقن بکل من التتراسیکلین والانروفلوکساسین إلی تقصى غیر معنوی لکل من العدد الکلى لکرات الدم الحمراء والبیضاء ، فی حین  تقصت معنویا الصفائح الدمویة بالمعاملة التتراسیکلین والأنروفلوکساسین. بینما لم یتأثر کل من الهیموجلوبین والـPCV  بالمعاملة التتراسیکلین والأنروفلوکساسین. کما أدى استخدام التتراسیکلین والانروفلوکساسین إلى تقص معنوی لکل من البروتینیات الکلیة والبیلیروبین والکریاتین مقارنة بمجموعة الکنترول بینما لم یتأثر کل من الألبومین والجلوبیولین ونسبة الألبومین /الجلوبیولین.

 أدت المعاملة بالتتراسیکلین غلی حدوث نقص معنوی علی مستوی 5% فی مستوى نشاط إنزیم الکبد AST وزیادة معنویة فی مستوى نشاط أنزیم الکبد ALT  بینما الحقن بالانروفلوکساسین أدى إلى تقص معنوی فی مستوى نشاط أنزیمات الکبد ALT & AST  مقارنة بمجموعة الکنترول.

أوضح الفحص المیکروسکوب أن نسیج کل من الکبد والکلى والخصیتین محتفظ بطبیعة دون أی إصابات مرضیة ولا یوجد أی  تأثیرات غیر مرغوب فیها للمعامله التتراسیکلین والانروفلوکساسین.  تحسنت بصورة معنویة کل الصفات الطبیعیة للسائل المنوی على مستوی احتمال 1% فی الأرانب المعاملة التتراسیکلین والانروفلوکساسین مقارنة بمجموعة الکنترول.

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