This thesis investigated the effect of cadmium sulfate exposure and treatment with the mushroom Pleurotus florida on albino rats. Rats were divided into groups that received various doses of cadmium and mushroom extract. Organs and blood were analyzed after 4, 8, 12, and 16 days. Histological analysis found damage to heart, liver, and kidneys in cadmium-exposed rats, which was reduced by mushroom treatment. Behavioral changes and clinical signs of toxicity were also observed with cadmium exposure. The mushroom extract showed protective effects on the organs. In conclusion, Pleurotus florida has potential for reducing cadmium-induced organ damage.

1. TITLE OF THESIS:
Effect of Mushroom (Pleurotus florida) on the Haematology and histopathology of Albino rat
(Rattus norvegicus) exposed to cadmium sulphate
Abstract
The abatement properties of mushroom on cadmium-induced damage in albino rat
organs was investigated. Male and female Wistar albino rats were divided into
control, and treatment groups. The study was conducted for thirty seven (37) days.
After acclimatization for three weeks. The rats were randomly divided into 7
groups of four rats each (2 males and 2 female). The animals were treated with
cadmium and mushroom for an experimental duration of sixteen days. Group 1 to
3 were fed orally with aid of gavage with 26mg/kg of the sclerotia extract daily and
2.1g/30ml, 1.05g/30ml and 0.525g/30ml body weight respectively then group 4 to
6 were fed 2.1g/30ml, 1.05g/30ml, 0.525g/30ml cadmium sulfate and grower mash
feed respectively without sclerotia while 7 group was feed with grower mash and
portable drinking water only, this serve as the control. A rat from each group (same
sex) was sacrifice every fourth day and organ such as heart, kidney, liver were
harvested, weighed using AccuLab® USA, model v-303 electronic analytical
weighting balance. Haematological samples were also collected immediately from
abdominal aorta and from the heart using a 5ml disposable sterile syringe (each rat

2. to each syringe). Organs (heart, liver, kidney) were weighed and observed and
tissue section taken for histopathology. All sample were analyzed at the University
of Benin Teaching Hospital (UBTH) laboratory department. Aspartate
aminotransferase (AST) activities were monitored and histological examination
was carried out. Results indicate that cadmium-induced rats had significantly
increased relative weight of liver, heart and kidney when compared to controls.
The mean highest value of ±0.920 and lowest mean value of ±0.823 was recorded
for heart in group 4 and group 3 respectively. For kidney weight, highest mean
value of ±1.313 and lowest mean value of ± 0.790 was recorded for group 5 and
group 1 respectively. While liver recorded a highest mean value of ± 2.590 and
mean lowest value of ± 1.993 in group 1 and group 2 respectively. The weight
obtained from the different organs (liver, kidney, heart) of each treatment groups
was subjected to one way analysis of variance and the following significant
difference was observed; group 1, 3, 4, 6 shows very highly significant difference
while group 2 and 5 shows highly significant difference. In cadmium-induced rats,
serum AST levels shows no significant difference compared with the control and
the abatement. Furthermore, histological alteration in liver, kidney and heart where
observed in different treatment groups (male and female) and this alterations where
moderately reverted with the pleurotus florida as abatement. Haematological
results in this study shows variations in different treatment groups and seems to
bear no direct relationship with the treatment. There was behavioral and physical
changes which ranged from swollen stomach, dimmed eyes, rusty fur,

3. hyperactivity, less response to stimuli, loss of appetite and aggressiveness. This
was not observed in the control and declined progressively in group 1, 2, 3 after the
first 4 days of administration of pleurotus florida extract. In conclusion, the extract
indicates hepatoprotective properties that eliminate the deleterious effects of toxic
metabolites of cadmium as shown in reversion of patchy vascular intimal erosion
of the heart, reversion of mild vascular hypertrophy of the liver and patchy cloudy
swelling (tubular necrosis) to fairly normal tubules of the kidney in both males and
female counterparts.

4. Aims and Objectives
The aims of this study was to evaluate the effect of different doses of cadmium
sulfate on the heart, kidney, liver, and some hematological parameters of wista
albino rats and the level of abatement of different concentration of mush room
(Pleurotus florida) to this effect. In line with this aim, this study was design to
investigate the following objectives which include the following:
1. The behavioral and physical changes in albino rats exposed to cadmium
2. The structural change in the histology of the heart, kidney, liver
3. Hematological changes in the treated groups
4. To determine the level of serum aspartate amino transferase
5. To ascertain the ameliorating properties of mushroom (Pleurotus florida)

5. METHODOLOGY
3.2.1 Study Location
The study was carried out at the animal unit of the Animal and Environmental
Biology Department, faculty of life science, university of Benin.
3.2.2 Experimental Animal
A total of twenty eight albino rats (Rattus norvegicus) of about (6 – 7 week old)
with average body weight of 236g (14 males and 14 female) obtain from the
animal house faculty of pharmacy, university of Benin, Nigeria and acclimatize for
two weeks on poultry feed pellet (grower mash feed) produce by Grand cereals
limited with portable drinking water given ad libitum at room temperature within a
twelve hours light and dark cycle before the commencement of the study.
3.2.3 Sample
Cadmium sulfate (CdSO4) with LD50 of 280mg/kg body weight was used for the
study. The metal concentration of the stock solution of 7%, 3.5% and 1.75% used
in the study was prepare by dissolving 2.1g, 1.05g and 0.525g of the cadmium in
30ml distilled water, equivalent to one fourth, one eight and one sixteenth of the
oral LD50.

6. 3.2.3.1 Abatement
The abatement use in this study was fresh sclerotia of mushroom (Pleurotus
florida) viability was tested via tissue culture in order to ensure that fresh sclerotia
(resting stage of some fungi) had not deteriorated in quality.
3.2.5 Experimental Design
The study was conducted for thirty seven (37) days. After acclimatization for three
weeks. The rats were randomly divided into 7groups of four rats each (2 males and
2 female). The animals were treated with cadmium and daily with mushroom for
an experimental duration of sixteen days. Group 1 to 3 were fed orally with aid of
gavage with 26mg/kg of the sclerotia extract daily and 2.1g/30ml, 1.05g/30ml and
0.525g/30ml body weight respectively then group 4 to 6 were fed 2.1g/30ml,
1.05g/30ml, 0.525g/30ml cadmium sulfate and grower mash feed respectively
without sclerotia while 7 group was feed with grower mash and portable drinking
water only, this serve as the control. A rat from each group (same sex) was
sacrifice every fourth day and organ such as heart, kidney, liver were harvested,
weighed using AccuLab® USA, model v-303 electronic analytical weighting
balance. Hematological samples were also collected immediately from abdominal
aorta and from the heart using a 5ml disposable sterile syringe (each rat to each
syringe). All sample were taken to University of Benin Teaching Hospital (UBTH)
laboratory department for analysis. The organs (heart, kidney, liver) were weigh
and observe and tissue section taken for histopathology.

7. 3.10 Clinical Observation
Each rats in each of the treated group was observed daily (evening after exposure)
for signs of clinical toxicity in the appearance of the fur, emaciation, decrease in
weight, swollen stomach, hyperactivity, nervous disorder, less response to stimuli,
loss of appetite.
3.2.6 Collection of Sample
3.3.0 Haematological Analysis
At the fourth day after exposure, the rats were placed in a chloroform chamber
(cotton wool soaked with chloroform) this made the rats inactive and then
sacrificed by cervical dissection. Blood was collected from the abdominal aorta
and from the heart using a 5ml disposable syringe and transfer into 2 samples
bottles, the first contain EDTA for complete blood count (CBC) analysis and the
second contain fluoride oxalate. Blood sample where allowed to clot, centrifuged
at 300rpm for 20 minutes to obtain a clear serum and then stored at -700
C prior to
biochemical analysis within 48hours.
3.3.1 Full Blood Count Analysis
Primary blood indices (Red blood cell count, white blood cell count, lymphocyte
count, haemoglobin value, mean corpuscular volume) were obtained for complete
blood count analysis by Sysmex (R) XT – series automated haematology analyzer.

8. 3.2.6 Histopathology
Organs such as liver, kidney and heart from exposed and control animals were
fixed in 10% neutral buffered formalin. After 48hours of fixation, organs were
dehydrate by passing through ascending order were dehydrate by passing through
ascending order of ethyl alcohol water concentrations, cleared in xylene and
sequentially embedded in paraffin wax blocks using rotary microtome. Tissue
sections of 3 - 5µm thick were cut, stained with haematoxylin - eosin (H & E) then
mounted in neutral DPX medium for morphological evaluation before microscopic
examination at X 400.
3.2.7 Statistical Analysis
Statistical analysis was done using general descriptive statistics and one way
analysis of variance at the P < 0.05 significant level, significant difference were
defined at P < 0.05. Computer software was used for statistical analysis ( SPSS).

9. 4.0 Findings
1. Clinical toxicity signs observed in treated rats include emaciation, swollen
stomach, dimmed eyes with deep pink/red coloration.
2. Histopathological damage in the cadmium exposed rats include vascular
hypertrophy, patchy intimal erosion and focal vascular intimal erosion,
atrophied follicle, cloudy and focal tubular swellings. This was not observed
in the control group.
3. Haematological results in this study shows variations in different treatment
groups and seems to bear no direct relationship with the treatment.
4. Behavioral changes observed in treatment rats during the course of the study
include hyperactivity, less response to stimuli, loss of appetite, fierceness
and aggressiveness especially in group 4, 5 and 6. Restiveness was also
observed in treated groups.
5. Mushroom (Pleurotus florida) shows abatements properties for the different
organs (heart, liver, kidney) as observed in the different group of both male
and female rats treated with the extract.
6. There was no significant difference obtained in the serum arspartate level of
the different treatment groups

10. Contribution to knowledge
• Data on histological parameters of rats expose to cadmium have been
obtained
• Bioinformatics on the effects of cadmium on organs of rats have been
provided
• Information on biochemical response of rats tissues to cadmium have
been provided
• This study is relevant in environmental waste management and in
regulations of chemicals
• This findings will be sent to local and international journal for
publication and more study encouraged to ascertain the impact of
pleurutus florida on organ damage

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