Effect of Cefotaxime Administration on the kidney, liver and Lung of Swiss white Mice (mus musculus)

The present study was designed to monitor the impact a drug such as cefotaxime has on the histological architecture of the kidney, liver and Lung on Swiss white mice. The experiment was designed by using 30 Swiss white mice from both sexes. - Mice were classified randomly into three groups:


Introduction
CLAFORAN (cefotaxime) is a semi-synthetic antibiotic widely used for different types of inflammation.CLAFORAN solutions range from very pale yellow to light amber which depends on the concentration and the diluent used.The pH of Cefotaxime solutions ranges from 5.0-7.5 slightly acidic to neutral composition (1).Claforan is supplied as a dry powder in appearance .It is a broad-spectrum antibiotic with activity against numerous gram-positive and gram-negative bacteria in the body and comes as a solution for injection to be administered by a healthcare provider into the vein or muscle.Hoechst-Roussel was the Pharmaceuticals that developed this drug in the late 1980s, It is on the World Health Organization's List of Essential Medicines, the most important medications needed in a basic health system (2).A variety of infections are treated with Cefotaxime such as:  Lower respiratory tract infections  urinary tract infections  Gynecologic infections  Bacteremia/septicemia  Intra-abdominal infections  Bone and join infections  Central Nervous System Infections  Skin Infections (3)(1) Cefotaxime antibiotic is active against many Grampositive and Gram-negative bacteria.Cefotaxime typically is effective against these organisms (in addition to many other).(4)As many drugs common side effects or reactions experienced can be found such as:  Pain and inflammation at the site of injection (4.3%)  Rash, pruritus, or fever (2.4%)  Colitis, diarrhea, nausea, vomiting (1.4%)(5) Cefotaxime, such as other anti-infective drugs, may cause irritation to tissues, It could result in tissue damage and require surgery to treat its effect.Leukopenia, neutropenia, granulocytopenia and even other rare affects such as pancytopenia, or agranulocytosis may develop during treatment with injected Cefotaxime.(6) Physicians and veterinarian should be aware to the alterations caused by these drugs while treating infections such as changes in some biochemical parameters, to avoid incorrect diagnosis.This study in addition to other Studies mentioned in this research has shown that chronic administration of cefotaxime in high doses may cause severe damage to the kidney, Liver and Lung.

Aim
The aims of this study are: 1.To observe and show the histological changes on certain tissues and cells in specific organs such as the Kidney, Liver and Lung of mice treated with Cefotaxime.

Experimental Design
The mice were divided randomly into three groups, two groups were injected intramuscularly in the thigh with Cefotaxime powder dissolved in distilled water to be injected directly.Mice were injected daily for 6 days in a row at 4 pm.Group A mice were injected with 0.5 ml of distilled water in the same manner, while Group B were injected with 0.1 ml and Group C with 0.2 ml (7).

Histological technique
The mice were killed using chloroform in a closed glass box to insure no ventilation.Kidney, Liver and Lung were obtained after dissecting the animal under aseptic conditions.

Histological technique carried out according to Brancroft &Stevens (1987) method as following: 1. Fixation
Organs after removal were fixed in 10% neutral buffered formalin solution (10ml of 40% formaldehyde + 90 ml tap water) for 22 hours at room temperature.

Dehydration
Organs were removed from the formalin and washed under running tap water for about 30 min.Tthen passed through progressively graded concentrations of alcohol baths (50%-70%-80%-90 and 100%). 1 hour in both 80% and 90% whilst 1/2 hour in two changes of absolute alcohol.

Clearing
Two changes of Xylene for 30 min.each, to insure the removal of any traces of alcohol.This led to some degree of transparency.

Infiltration and Embedding
Tissues were passed a mixture of xylene and molten parrafin wax (58 o C) for 30min.Later stainless steel containers were used for creating labeled blocks.

Tissue Attachment
Mayer's glycerol-albumin was smeared over the slides this mixture was used as a preservative agent.

Staining
Slides were stained with Heamatoxylin for about 10 min.then washed with running tap water then transferred into Eosin Stain for 1 min.Dehydration in ascending concentrations of 5min.alcohol baths (50%-70%-80%-90%-100%-100%).Slides were finally cleared with xylene.9. Mounting DPX was used as a mounting agent, and then covered with cover slips.

1) Kidney
-Control Kidney The glomerulus because small in size and the space between the glomerulus and the capsule was very wide, this indication referred to atrophy of glomerulus as in Fig ( 3), in some regions the glomerulus are segmented as in Fig ( 3).Many alterations happened to urinary tubules in both proximal and distal tubules including unclear cells arrangement that line the tubules which usually are cuboidal in shape, but here the cells appeared with necrotic cytoplasm and degenerated nucleus (pyknosis, karyorrhexis and karylysis) in additional to broken of urinary tubules and desquamation of epithelial from the basement membrane and pushed towards the lumen of the tubules with formation of cast in some urinary tubule lumen most epithelial cells become short columnar or columnar causing reducing the lumen of the urinary tubules.
-Extra Therapeutical Dose for Kidney The presence of hyperplasia can be recognized in this dose, which caused thickening of the alveolar wall that composed simple squamous epithelia at normal conditions.This thickening caused a reduction of the alveoli lumen and caused also accumulation of mucous material on the surface of the cells of the alveoli.Sometimes hemorrhage can be seen in certain places in the lung but other parts showed normal cell shape.
-Extra Therapeutical Dose for Lung The increase and thickening of the wall was noticed and additional cells (epithelial cells), hyperplasia and infiltration of lymphocytes that formed a thick wall in the stroma on the lung.

Discussion
Claforans antibacterial action results from inhibition of cell wall synthesis.It is stable against the action of most β-lactamases.Due to its attack mechanism on bacterial cell wall synthesis, β-lactams are considered to be bactericidal.(1)(4) Cefotaxime could be intramuscular injected (IM) or intravenous infused (IV).Doses should be adjusted according to the patient's renal or hepatic impairment due to cefotaxime metabolization to active and inactive metabolites by the liver and excreted in the urine in very high levels.(8) This is an indication to the high impact this drug has on both kidney and liver in the body and an explanation for its huge effect on their cells.Cefotaximes histopathological effects on the kidney, Lung and liver were shown in the Results.The liver and kidney showed vacuolar degeneration of the hepatocytes with the appearance of infiltration of the lymphocytes and necrosis of the hepatocytes.The kidney showed inter-tubular hemorrhage with necrosis of some renal tubules, this was in correlation to a study (11) showing similar results found in both the kidney and liver of adult rats treated with Cefotaxime over 7 days.Other studies (12)(13) also found that ceftriaxone therapy associated with elevated liver enzymes in rats after IM injections and obtained biochemical changes attributed to damage to hepatocytes and renal tubule cells, these biochemical changes and results obtained was in agreement with this study.The histopathological findings in kidneys of cefotaxime treated mice obtained in this study were consistent with those obtained in a study ( 14) that observed cephaloridine administration to rats caused degeneration and necrosis of the renal proximal tubular epithelia.Finally another study (15) observed the drugs alteration on the renal proximal tubular causing changes such as necrosis, hyaline cast and calcification, suggesting renal disorders, this was in correlation with the present study as well and shows the great impact of Cefotaxime on a cellular level or organization.

Conclusion
In conclusion, drug utilization such as cefotaxime has a great impact on the cells of liver, kidney and Lung causing cells and tissue structure to be altered severely.
Cetotaxime showed its effect on many body organs and tissue so it should be taken in consideration that such drug should be taken according to age and type of inflammation in the body to protect them from its significant impact.Cefotaxime had caused hepatic and renal histopathological cellular alterations; hence Cefotaxime should not be taken by choice whether used on animals or patients suffering from hepatic and renal disorders.