Research Article

Growth Performance, Blood Profile, Organosomatic Indices and Histopathology of Clarias gariepinus Fed Amoxicillin as Dietary Additive  

Adewole AdeyemoMuniru
Department of Animal and Environmental Biology, AdekunleAjasin University, PMB 001, AkungbaAkoko, Ondo State, Nigeria
Author    Correspondence author
International Journal of Aquaculture, 2016, Vol. 6, No. 15   doi: 10.5376/ija.2016.06.0015
Received: 15 Aug., 2016    Accepted: 10 Nov., 2016    Published: 28 Dec., 2016
© 2016 BioPublisher Publishing Platform
This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:

Adewole A.M., 2016, Growth performance, blood profile, organosomatic indices and histopathology of Clarias gariepinus fed amoxicillin as dietary additive, International Journal of Aquaculture, 6(15): 1-13 (doi: 10.5376/ija.2016.06.0015)

Abstract

A 12 week feeding trial was conducted to investigate the effect of administration of different dosages of amoxicillin on biophysiological responses: nutritional, haematological, serum biochemical, organosomatic indices and histopathological study of juvenile Clarias gariepinus. One hundred and eighty juvenile C. gariepinus with the mean weight of 10.94 ± 0.01 g were stocked into the different treatments using a completely randomized design. The fishes were fed diets containing iso-nitrogenous crude protein (40%) at 5% body weight, twice a day. Histopathological results showed no visible lesions in some of the tissues of the fish fed with control and amoxicillin diets. However, there were marked pathological conditions that were manifested in some organs like the liver, brain, kidney and intestine of fish. The inclusion of amoxicillin at 0.5% in the diet of C. gariepinus is being advocated in sustainable catfish production in Nigeria.

Keywords
Amoxicillin; Biophysiological; Clarias gariepinus

1 Introduction

Antibiotics are extensively used in human and veterinary medicine with or without regulations mostly among small scale farmers, with a resultant emergence of multidrug resistance bacteria. The type and amount of antibiotics used in aquaculture depend on the farming practices, local and national regulations, as well as government enforcement policies. The restrictions on the use of antibiotics are different with some countries recently introducing stricter regulations in response to the global threat of antimicrobial resistance and consumer concerns about residues in food. The majority of aquaculture productions, however, take place in countries with “permissive regulations” and limited environmental monitoring. The overall use, therefore, varies widely between countries, ranging from 1 g/metric ton of production in Norway to 700 g/metric ton in Vietnam (Defoirdt et al., 2011).

 

Among the class of antimicrobial agents used for growth promotion in Nigeria. tetracyclines, penicillins, macrolides, lincomycin and fluoroquinones are the most commonly used antimicrobial agents in animals for treatment and prevention of diseases. Evaluating the quantity of antimicrobial agents and how they are used in animal production in Nigeria is an important issue to be considered, because studies have shown that animal drugs are grossly misused in Nigeria and are readily available without prescription (Aibinu et al., 2007; Olatoye and Basiru, 2013; Olufemi et al., 2015) for prevention of disease outbreak or to reduce mortality losses without engaging the services of veterinarians for disease diagnosis and treatment.

 

Aquatic animals are colonized by trillions of microorganisms that have a symbiotic relationship with their hosts and are distributed in gills, body surface and gastrointestinal tract and played an important role in nutritional, physiological and pathological events (Denev et al., 2009). Antibiotics are drugs of natural or synthetic origin that have the capacity to kill or inhibit growth of microorganisms (NAOH, 2001). Antibiotic growth promoters (AGPs) are administered at low doses in the feed and they act by specifically reducing the number of pathogenic bacteria in the gut (Dafwang et al., 1987). Supplementing animal feed with antimicrobial agents to enhance growth has been a common practice for more than 30 years and it is estimated to constitute more than half of the total anti- microbial drugs used worldwide Ereke et al. (2015).

 

Antimicrobial drugs have the potential to positively affect growth of animals through better appetite, improved feed conversion, stimulation of immune system and increased vitality and regulation of the intestinal microflora. Furthermore, antimicrobial growth promoters (AGPs) in animal nutrition have been of beneficial effect in improving growth performance and prevention of diseases (Barton, 2000; Snel et al, 2002; Adewole, 2016). Recently, several antibiotics have been used indiscriminately due to accelerated growth of aquaculture in Nigeria, while several researches have been done to evaluate the appropriate inclusion levels of some common antibiotics used in fish production. However, in a separate study, Adewole (2016) evaluated the growth rate of catfish Clarias gariepinus to different inclusion levels of oxytetracycline, while Lawal et al. (2012) determined the inclusion levels of oxytetracycline and furasol as antibiotics, respectively. But, there is dearth of information on the inclusion levels of amoxicillin in the diet of Clarias gariepinus in Ibadan, Nigeria. This study, therefore, sought to evaluate the effect of different doses of amoxicillin on the growth performance, haematology, organosomatic indices and histopathology of Clarias gariepinus.

 

2 Materials and Methods

Study Area: The experiment was carried out in the Department of Aquaculture and Fisheries Management, University of Ibadan, Ibadan, Nigeria.

 

Formulation and preparation of experimental diets: The experimental diets were formulated using algebraic method according to Falayi (2003) as shown in Table 1. The diets were prepared with amoxicillin at the inclusion levels of (0.00%) AMX1 (control without antibiotic), (0.10%) AMX2 and (0.5%) AMX3 based on the reported dosages of antibiotics by Crevadi et al. (1987). After the formulation, the different ingredients were weighed using the electric sensitive weighing balance (OHAUS-LS 200 Model). The dough made after was pelleted, sun dried, packed in tight plastic bags, labeled accordingly and stored at room temperature in the laboratory until its ready for use as reported by Adewole (2016). Different batches of the feed were prepared every month throughout the duration of the experiment as recommended by (Sogbesan, 2007).

 

Table 1 Percentage composition of ingredients (g/100 g diets) in Amoxicillin meal diets for feeding trial

Note: AMX =Amoxicillin

 

Stocking of experimental fish and Experimental design: The healthy juvenile of Clarias gariepinus (n=20, mean weight 10.94 ± 0.01 g were randomly sorted, weighed and stocked into the experimental tanks in triplicates using a completely randomized design as reported earlier by Adewole (2016), after been acclimatized for 2 weeks and the fish were feed at 5% body weight twice daily at 8.00 and 17.00 hours for 84 days.

 

Sampling and Data collection: The length and weight of fish in each tank was measured to determine the different rearing indices such as: mean weight gain, specific growth rate, relative growth rate, nutrient utilization parameters, condition factor and survival rate by Ajayi et al. (2013) and Adewole, (2016).

 

 

Where:

W0 = initial mean weight

W1= final mean weight

 

 

Where:

Ln = Natural log

W0 = initial mean weight

W1 = final mean weight

 

 

 

Where:

W = final weight

L = Final standard length

 

 

Where:

NI = final number of fish at the end of experiment

N0 = initial number of fish at the beginning of experiment

 

 

 

Haematological and serum biochemical study

The blood collection was done before and after the feeding trials. Fish samples were selected from the various treatment tanks and blood was collected through the caudal vein using a hypodemic needle into 2 sets of sample bottles. One set contained ethylene diamine tetra acetic acid (EDTA) as anticoagulant, which was used for haematological analysis according to Ajayi et al., (2013), while the second set without EDTA was used for serum biochemical analysis as earlier described by Adewole and Gbore (2006).

 

Organosomatic and Histopathological study

Histopathological analysis was carried out following method of Roberts (1978): The following organs such as brain, liver, kidney, heart, and gill, intestine were dissected from the fish and placed in the sample bottles, 10% formalin was added to prevent tissue degeneration and labeled accordingly. Further analysis of the organs were carried out using Masson’s trichrome with Hematoxylin Eosin and Saffron (HES) stain on the organs to examine the general morphology. The tissues were examined in a Tecnal G2 Brotwin (FER) electron microscope using an accelerated voltage of 100kv. The magnification was taken at 400X as reported by Adewole (2016).

 

Statistical analysis:

The data generated were analyzed statistically using Analysis of Variance (ANOVA) at (α 0.05) and descriptive statistics following Statistical Analysis System software (SAS, 2008) model. The significant treatment means were compared using Duncan’s multiple range test (DMRT) of the same software.

 

3 Results

The fish fed the experimental diets did utilize the feeds at the varying levels, thus leading to the significant variations in some of the growth parameters recorded: Total final weight (TFW) attained ranged from 601.57 ± 21.23 g to 1 026.87 ± 57.19 g. The highest value was from fish fed AMX2 diet and the lowest value was from the fish fed the control diet. The fish fed the control diet was significantly lower (p < 0.05) in total final weight attained compared to other diets. There was no significant difference in the TFW recorded between the fish fed AMX2 and AMX3 diet (Table 2).

 

Table 2 Growth performance, nutrient utilization and survival rate of Clarias gariepinus fed Amoxicillin meal diets (AMX’s)

Note: Values with different superscripts in each row are significantly different (P > 0.05)

 

Mean final weight (MFW), total weight gain (TWG), mean weight gain (MWG), specific growth rate (SGR), relative growth rate (RGR) all ranged from (37.60 ± 1.33 g to 59.88 ± 6.22 g, 382.90 ± 20.80 g to 808.20 ± 57.62 g, 23.93 ± 1.30 to 56.39 ± 1.71 g, 0.52 ± 0.02 to 0.80 ± 0.03% / day). The highest MFW and TWG values from the fish fed AMX3 and AMX2 diets respectively were significantly different (p < 0.05) from the fish fed AMX1 diet, while the lowest value was from the fish fed (AMX1) control diet (Table 2). The highest MWG value of 47.02 ± 6.20 g was from fish fed AMX3 diet, the lowest value of 23.93 ± 1.30 g was from fish fed the control diet. The fish fed the control diet had significantly lower (p < 0.05) MWG compared to other diets. There was no significant difference in the MWG of fish fed AMX2 and AMX3 diets. Highest SGR value of 0.80% / day was from fish fed AMX2 diet and the lowest value of 0.52 ± 0.02% / day was from the fish fed control diet. The fish fed the control diet had significantly lower (p < 0.05) SGR compared to others. There was no significant difference (p < 0.05) in the SGR between the fish fed AMX2 and AMX3 diets (Table 2). The RGR of the fish fed the experimental diets followed the same trends as SGR.

 

The final condition factor (K2) had the highest value of 0.48 ± 0.03 from fish fed AMX3 diet and lowest value of 0.39 ± 0.09 from fish fed AMX2 diet. There was no significant difference in the K2 of all the fish fed the experimental diets (Table 2). The percentage survival of the fish fed the tested diets significantly (P < 0.05) ranged from 80.00 ± 0.00 to 90.00 ± 0.00%. The highest value was from the fish fed AMX2 diet and lowest value of 80.00 ± 0.00% was from fish fed the control diet.

 

The total feed intake (TFI) ranged from 1701.18 ± 27.69 – 2362.95 ± 64.19 g. The TFI was significantly different among the treated diets, with the highest value of 2362.19g from the fish fed AMX3 and the least 1701.18 ± 27.69 g from the fish fed AMX1 (Control). The mean feed intake (MFI), total protein intake (TPI) and mean protein intake (MPI) also varied significantly within the treatment and followed the same trend as the TPI in the experiment. The feed conversion ratio (FCR) followed the opposite trend of the TFI, MFI and MPI. The highest FCR was from the fish fed the control, while the lowest was from fish fed the AMX2 diets. The protein efficiency ratio (PER) was significantly different among the treatments. The highest value of 0.87 ± 0.00 was from the fish fed AMX2 and the least value of 0.54 ± 0.01 was from the fish fed AMX1 diet (Table 2).

 

The pack cell volume (PCV) values ranged from 22.33 ± 0.88 to 27.67 ± 1.45%. The highest value was from the fish fed AMX2 diet, followed closely by 26.67 ± 1.89% from AMX3 diet and the lowest value was from the control diet. All the final PCV values were higher than the initial (Table 3). The fish fed AMX2, had significantly (P < 0.05) higher PCV than AMX1 (control) diet but differed insignificantly. (P > 0.05) from fish fed AMX3 diet (Table 3). The red blood cell hemoglobin (Hb) from the fish fed all experimental diets followed the same pattern as PCV. The red blood cell (RBC) counts ranged from 11.64 ± 1.49 to 17.47 ± 1. 57 x 106/dl. The highest RBC counts value was from the fish fed AMX3 diet, followed closely by 14.57 ± 0.58 from AMX2 diet, while the lowest value was from the fish fed control diet (AMX1). All the final RBC counts were higher than the initial, the fish fed AMX3 diets had significantly (P < 0.05) higher RBC counts than the control (AMX1), but differed insignificantly (p > 0.05) from the fish fed AMX2 diets (Table 3). The mean cell volume (MCH) parameter of the red blood cell ranged from 63.00 ± 3.00 to 65.67 ± 2.85 x104/l. The highest value was from the fish fed AMX1 diet, followed closely by 65.33 ± 1.45 x104/l from the fish fed AMX2 diet, while the lowest value was from the fish fed AMX3 diet. All the final MCH were higher than the initial value of 55.00 ± 7.00 x104/l. There was no significant difference (P > 0.05) in the MCV between the fish fed the control diet and other diets (Table 3). All the final MCH and MCHC values were higher than the initial values except the fish fed AMX3 diet. There were no significant difference (P > 0.05) in the MCH and MCHC values between the fish fed the control diet and fish fed the other diets (Table 3).

 

Table 3 Haematological parameters of Clarias gariepinus fed amoxicillin meal diets

Note: Values with different superscripts in each row are significantly different (P > 0.05)

 

The white blood cell (WBC) counts ranged from 8.87 ± 1.45 to 11.82 ± 0.70 x103/dl. The highest value was from the fish fed control diet, followed closely by 9.20 ± 0.35 x103/dl from fish fed AMX2 diet, while the lowest was from the fish fed AMX3 diet. All the final WBC counts were higher (P < 0.05) than the initial. The fish fed the control diet, had significantly higher (P < 0.05) WBC counts than others (Table 3).The erythrocyte sedimentation rate (ESR) varied from 2.00 ± 1.00 to 2.67 ± 0.67 mm/hr. The fish fed AMX3 had the highest value for ESR, while the lowest value was from the fish fed AMX2 diet. The final ESR was higher than the initial and there was not significantly difference (P > 0.05) among the ESR value of the fish fed the experimental diets (Table 3).

 

The mean final total protein for the fish fed the experimental diets ranged from 6.30 ± 0.15 g/100 ml to 7.27 ± 0.09 g/100 ml. All the final total proteins levels were higher than the initial value of 5.83 ± 0.007 g/100 ml.The highest total serum proteins was from the fish fed AMX3 diet and the lowest value was from the fish fed AMX1 diet. The fish fed the tested diets had significantly (p<0.05). (p<0.05) higher mean final total protein than the fish fed the control diet (Table 4).The mean final albumin and globulin followed the same trend as the total protein in the fish fed the experimental diets (Table 4).

 

Table 4 Serum biochemical indices of Clarias gariepinus fed amoxicillin meal diets

Note: Values with different superscripts in each row are significantly different (P > 0.05)

 

The mean final serum blood glucose values ranged from 45.67 ± 1.45 g/100 ml to 51.33 ± 1.20 g/100 ml. All the final mean blood glucose values were higher than the initial value (35.67 ± 0.88 g/100 ml). The highest value was from the fish fed AMX2 diet and the lowest value was from fish fed AMX3 diet. The fish fed the AMX2, AMX1 had significantly higher (p < 0.05) mean blood glucose than AMX3 diets (Table 4).

 

The mean final aspartate aminotransferase (AST) values ranged from 18.33 ± 3.38 to 29.67 ± 3.18 µ/l. All the mean final blood AST values were higher compared to the initial value 20.00 ± 2.31 µ/l except AMX1 diet. There were no significant differences between the highest and lowest final blood AST values of all the fish fed AMX’s diets (Table 4). The mean final alanine aminotransferase (ALT) values ranged from 61.00 ± 2.08 to 87.33 ± 1.76µ/l. All the fish fed AMX’s experimental diets were higher in ALT compared to the initial value of 72.00 ± 6.43 µ/l except AMX1. The highest value was from fish fed AMX3 diet while the lowest was from AMX1 diet. The fish fed the AMX3 and AMX2 had significantly higher (p < 0.05) mean ALT than AMX1 diet. The blood final mean ALP ranged from 35.33 ± 0.67 to 42.67 ± 1.17 µ/l. The highest value was from AMX3 diet, while the lowest was from the fish fed AMX1 diet. Fish fed AMX3 diet had significantly higher (p < 0.05) ALP than the fish fed AMX2 and AMX1 diets (Table 4).

 

Cardiosomatic indices for the fish fed the experimental diets ranged from 0.10 ± 0.01% to 0.21 ± 0.02%. The fish fed AMX1 (control diet) had the highest and lowest values were from the fish fed AMX2 diet. The fish fed the control diet had significantly higher (p < 0.05) cardiosomatic index compared to others (Table 5) Hepatosomatic indices for the fish fed the experimental diets ranged from 0.87 ± 0.19% to 1.81 ± 0.39%. The highest hepatosomatic index value was from AMX2 diet and the lowest value was from the fish fed control diet. The fish fed the control diet had significantly lower (p<0.05) hepatosomatic index compared to others, while no significant difference (p<0.05) existed between fish fed AMX2 and AMX3 diets (Table 5). Renosomatic indices for the fish fed the experimental diets ranged from 0.21 ± 0.02% to 0.49 ± 0.01%. The highest value was from fish fed AMX2 diet, while the lowest value was from the fish fed the control diet. The fish fed the control diet has significantly lower (p<0.05) hepatosomatic index than AMX2, but not significantly different from AMX3 diet (Table 5).

 

Table 5 Organosomatic indices of Clarias gariepinus fed Amoxicillin meal (AMX’S) diets

Note: Values with different superscripts in each row are significantly different (P < 0.05)

 

The histopathological examinations varied with degree of lesions from mild to severe in the various organs of the fish fed the tested diets as shown in Table 6.

 

Table 6 Summary of histopathology of the tissues of Clarias gariepinus fed Amoxicillin meal (AMX’s) diets and Control diet

 

4 Discussions

The feeding trials revealed that C. gariepinus responded to all the diets, irrespective of amoxicillin concentrations. Amoxicillin promoted the growth of C. gariepinus. C. gariepinus was able to effectively utilize the amoxicillin for growth. The highest growth performance was observed in the fish fed AMX2 diet, although this was not significantly different from the fish fed AMX3 diet. This better growth performance in fish fed the antibiotic may be attributed to the levels of antibiotic in the diet which might have destroyed pathogenic organisms in the alimentary canal, thus reducing morbidity and mortality due to reduced sub - clinical diseases by Rosen (1995). Furthermore, the use of antibiotics have been reported to enhance digestibility by preventing the irritation of the intestinal lining and effective utilization of the nutrients from the intestine by thinning of the mucosal layer, which led to improve animal performance and increased economic output (Dafwang et al., 1987; Ewing and Cole, 1994; FAO, 2005). The results from the present study agree with Shalaby et al. (2006) that there were no significant differences in the final weight gains among the fish fed chloramphenicol at 15, 30 and 45 mg/kg diets. Similar results have been obtained by Lawal et al. (2012) and Adewole (2016) that reported similar higher growth rate than the control in C. gariepinus fed varying levels of oxytetracycline and furasol as dietary additives.

 

The feed intake increased with increasing levels of AMX’s diets, but it was not significantly different among the treated groups but significantly differed from the control. This indicates that the C. gariepinus was able to accept and adapt to the diets/drugs even at higher level of inclusions and stimulate growth during the course of the trial. This observation was similar to the reported increase / higher feed intake and enhanced digestibility of the dietary components by the fish fed both tetracycline and furasol as dietary additives by Lawal et al. (2012).

 

Feed efficiency ratio (FER) and Protein efficiency ratio (PER) are used as quality indicators for fish diet and amino acid balance (Shalaby et al., 2006). The PER values from this study was similar to reported increased PER of O. niloticus fed varying levels of chloramphenicol as observed by Shalaby et al., 2006. Therefore the C. gariepinus was able to utilize efficiently the protein in the diet adequately or the diet provided the needed quality and quantity of protein for growth and health of the fish. The good overall growth performance of C. gariepinus fed AMX’s diets confirms the suitability of the chosen nutritional compositions for the fish. The feed conversion (FCR) in the present study was higher than values (0.73 ± 0.07 – 1.35 ± 0.10) obtained by Lawal et al., (2012), but similar to values reported by Adewole (2016). Furthermore, these values were better than those reported by El- Block (1975) that the FCR for Clarias ranged from 3.2 - 6.7, based on the quality of the feed. Furthermore, the better FCR recorded by the fish fed amoxicillin compared to the control group is in agreement with Anderson et al. (1999) who observed that the increase in nutrient uptake in broiler chickens consequently increased the growth of the animal.

 

Condition is a term widely used to refer to the overall physiological status of health of an individual (Cone, 1989; Stevenson and Woods, 2006) and it’s of central importance to many aspects of life history, including metabolism (Oskarsson, 2008), reproductive potential (Oftedal, 2000) and mortality (Dutil and Lambert, 2000). The condition factor in all the treatments reflected less than 1. Edward et al. (2010) expressed that a condition factor above 1 indicates better utilization of feeds by fish for growth and development. The result obtained in this study of less than 1 may be attributed to the prolonged culture period (84 days), which may have been accompanied with production stress.

 

Haematological profiles usually give vital information on the response of the body to injury, deprivation. Such an evaluation is indispensably important in arriving at a diagnosis, making prognosis and also in the assessment of efficacy of therapy and toxicity of drugs and chemical substances (Ihedioha et al., 2004). Therefore, the heamatological parameters of C. gariepinus fed to different AMX’s diet revealed that the PCV, RBC and ESR increased significantly within the tested diet, when compared to the control. The results obtained here were similar to the reported increase in blood parameters of O. niloticus fed graded levels of chloramphenicol (Shalaby et al., 2006) and Faisal (2003) reported the significant increase values of erythrocyte, haemoglobin and haematocrit of the same catfish C. gariepinus at 1st and 3rd day after administration of ciprofloxacin, amoxicillin and ampicillin.

 

The red blood cell indices (MCV, MCH and MCHC) are particularly important for the diagnosis of anaemia in most animals. This study showed a significant decrease of MCH, MCV in fish fed the highest level of Amoxicillin. This is similar to the reported decreased in MCH and MCHC of O. niloticus fed the highest level of chloramphenicol by Shalaby et al. (2006). These authors, therefore, assumed that decrease or increase of blood indices may be attributed to a defense reaction against drugs and diets, which occurs by stimulation of erythropoiesis.

 

The WBC counts in C. gariepinus fed different graded levels of amoxicillin diets decreased significantly when compared to the control. The increase in WBC (leucopoena/lencopomia) observed in the fish fed the control diet may be attributed to increase in production of leucocytes in the haemtopoietic tissues, such as kidney and spleen. Lymphocytes are the most numerous cells comprising the leucocytes, which function in the production of antibodies and chemical substances serving as defense against infection. The primary consequence of observed changes in leucocyte counts in stressed fish is suppression of the immune system and increased susceptibility to disease Wedemeyer and Wood (1974). Ahamefule et al. (2008) reported that decrease in number of WBC counts below the normal range is an indication of allergic conditions, anaphylactic shock and certain parasitism or presence of foreign body in circulating system. The values of WBC counts in this study were similar to (3 800 -7.800) and (3.41-10.89 GL-1) reported for the same fish species fed diet containing 0.1% of yeast, red Acalypha and tetracycline as additives (Osineye et al., 2015). Several factors have been reported to affect the haematological responses in fish, these include sex, age, size, environmental and physiological conditions (Sowumi, 2003; Adewole and Gbore, 2006).

 

Blood biochemistry can also be used as health indicator in fish and is important in assessment of the quality and suitability of feed ingredients in animals Satheeshkumar et al. (2011). Furthermore, Animashahun et al. (2006) affirmed that the comparison of blood chemical profiles with nutrient intake might indicate the need for adjustment of certain nutrients upward or down ward for different population groups. Iyayi and Tewe (1998) observed that serum urea, total protein and creatinine contents depend on both the quality and quantity of protein supplied in the diet.

 

Therefore, the effect of AMX’s diets on serum biochemistry of C. gariepinus presented a significant increase in both protein and albumin, while the globulin increments were not significant both within the tested diets and when compared with the control. The results obtained here were similar to Faisal (2003) that found serum total protein increased significantly in the C. gariepinus after the administration of both ciprofloxacin and amoxicillin antibiotics. On the other hand Shalaby et al. (2006) reported significant hyperproteinemia in all the fish groups administered with garlic, except in those that received (40 g garlic/kg) diet and the two highest levels of chloramphenicol (30 and 45 mg/kg diet).

 

Blood serum protein is a fairly labile biochemical system, precisely reflecting the condition of the organism and the changes to it under influence of internal and external factors. Booke (1964) showed that sex, spawning, food, osmotic pressure, temperature, light, age, hibernation, hormones, oxygen depletion, and season were factors that demand total serum protein compliment in fish. High serum protein levels have been reported due to improve liver and other organs functions which synthesized plasma protein (Metwally, 2009) while contrarily, high serum protein levels have been reported to be indicative of osmoregulatory dysfunction, haemodilution, or tissue damage surrounding blood vessels (Hille, 1982). Furthermore, the concentration of total protein in blood is used as a basic index for health, nutritional status and also reflection of altered metabolism in fish (Martinez, 1976; Gbore et al., 2016).

 

The blood glucose of C. gariepinus fed AMX’s diets revealed a significant reduction at the highest level of supplementation, while there was marginal non-significant increase in the plasma glucose concentration from the fish fed AMX2 diet when compared to control. The reduction in glucose concentration observed in this study agreed with Shalaby et al. (2006), who reported glucose reduction in fish fed diet containing the highest levels of Allium sativum (30 and 40 g/kg diet). These results were also similar to (Kumar and Reddy, 1999; Thomson and Ali, 2003) who found that feeding mice with (40 mg garlic/kg) body weight for 28 days induced significant decrease in the serum glucose levels. Contrarily, there were significantly elevated (p < 0.001) plasma glucose level in the O. niloticus administered with all doses of chloramphenicol, compared with those fed the control diet by Shalaby et al. (2006). Furthermore this result is in partial disagreement with those of Tarter (1986) and Faisal (2003), who verified that plasma glucose concentration in the same fish species increased significantly after oral administration of ciprofloxacin, amoxicillin and ampicillin. The difference with their positions may be due to the dosage and duration / length of the drugs usage, since longer duration and higher dosage in this study significantly reduced the plasma glucose concentrations. Increase blood glucose levels might have been due to glucose shift from tissues to blood or to an impairment of glucose mobilization. While, glucose concentration reduction in blood may be attributed to improving the antioxidant system in cell of pancreas to produce insulin (Metwally, 2009).

 

Plasma enzyme activities in fishes are considered to be a significant factor to assess the state of liver and some other organs in fish (Verma and Delela, 1981). Therefore, attention has been focused on the changes in AST, ALT and alkaline phosphatase (ALP) activities, which promote gluconeogenesis from amino acids, as well as on the changes in aminotransferase activities on the liver (Hilmy et al., 1981). Results of this study showed that serum enzymes ALT and ALP increased significantly in the fish groups fed amoxicillin. The increase in AST was however, insignificant. But insignificantly increased in AST activity was also observed. These results are well corroborated by significant increase in serum AST and ALT activities as reported earlier by the work of Saba et al., (2000), who administered chloramphenicol in rats, Bhat et al. (1998) on minocydine and Aubrecht et al. (1997) on hygromycin B. On the other hand AST and ALT activities decreased significantly in O. niloticus fed on all levels of Allium sativum and chloramphenicol (Shalaby et al., 2006). Celik (2004) observed that ALT and AST values can change due to the fish species. While Verma et al. (1994) and Asztalos et al. (1990) submitted that heavy metal compounds, pesticides, sublethal concentration of phenol and organic pollutants may induce changes in blood AST and ALT activities in fish.

 

The results of the organosomatic indices showed significant increase in the liver, kidneys, and insignificant increase in visceral somatic indices when compared to control. The enlargement of these organs might have resulted from deposition of some chemicals, which the fish was unable to metabolize (Sogbesan, 2007). However, the hypertrophy and hypotrophy of these organs has been associated with presence of toxin (Ewuola et al., 2003; Adewole and Awosusi, 2015). The hepatosomatic index values obtained in this study were higher than the values (1.64 ± 0.09%) obtained for the same fish species fed graded tetracycline meal diet by Adewole (2016). But these values were however similar to 1.54 – 1.70% recorded for O. niloticus fed different dosages of chloramphenicol as observed by Shalaby et al., 2006. Furthermore, the values reported in this study were lower to 6.11 – 7.30% reported by Haruna (2001) for C. gariepinus fed processed agricultural by products. However, the values obtained in the present study for the cardiosomatic and brainsomatic indices were lower than values (0.25 ± 0.01 and 0.87 ± 0.01%) reported for C. gariepinus fed graded tetracycline meal diets (Adewole, 2016).

 

There is a little or no information available about toxicity and pathological effects of the administration of amoxicillin in Clarias gariepinus. The result of the effect of feeding Clarias gariepinus with amoxicillin meal diets on the different organs of fish such as the kidney, heart, liver and intestine showed shrunken of glomerular tuft, thinning of myocardium, intracellular fat deposition and villi atrophy. The results of the toxicity of amoxicillin in this study were similar to the reported pathological effects of antibiotics in animal models such as acute tubular necrosis induced by gentamicin toxicity in rats by Hottendorf and Williams (1986); dog (Graucer et al., 1994); cat (Mealy and Boothe, 1994) and poultry (Khan et al., 2008). Furthermore, Acute Interstitial Nephritis (AIN) has been reported in a number of human patients who developed drug-related AIN while receiving intravenous amoxicillin therapy (Appel et al., 1981). Also tetracycline and furasol toxicities in C. gariepinus (Lawal et al., 2012; Adewole, 2016) resulting in conditions such as diffused vacuolation, with portal and central venous congestion, along prominent melanomacrophage - centres and vacuolation of glands cells / partly damaged glands with necrosis were reported.

 

5 Conclusions

It is interesting to note that better growth performance, nutrient utilization and survival rate were obtained with the fish treated with amoxicillin meal diets compared to control which was significant, but the observed hematological, serum biochemical and histopathological changes may indicate that the potentiality of amoxicillin to be toxic on prolonged administration from the evaluated dosages. Therefore, dose - response effect of amoxicillin on the organs and other physiological parameters of the fish at minimal concentration reflected improved growth with mild effect. Therefore, the inclusion level of amoxicillin at 0.1% of the feed may be well tolerated in the diet of C. gariepinus. Also the withdrawal period or plasma half-life for the drug should be evaluated to curb the prevalence of antibiotic residue in cultured fish in Nigeria.

 

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