ANALYSIS OF KIDNEY FILTRATION

Name : Pratiwi Kusuma Kurniawati

Student ID : B1B017007
Entourage : VI
Group :3
Assistant : Afif Ghalib Ammar Zain

PRACTICAL REPORT OF ANIMAL PHYSIOLOGY II

MINISTRY OF RESEARCH, TECHNOLOGY AND HIGHER EDUCATION

JENDERAL SOEDIRMAN UNIVERSITY
FACULTY OF BIOLOGY
PURWOKERTO
2019
 I. INTRODUCTION

A. Background

Excretion is the process of removing metabolic waste from the body which is
no longer needed by the body. Some toxic metabolic substances (poisons) for the body
include waste nitrogen. This nitrogen is produced when food is transformed into
energy. The nitrogen product is ammonia which is very toxic. Some animals convert
ammonia to urea or uric acid which is less toxic before being removed from the body
(Dahelmi, 1991). The predominant function of the kidney is excretion, which is
broadly defined as the removal of materials from the body. The broad category of
excretion can be subdivided into several more well-defined physiological processes
including nitrogenous waste elimination, ionic and osmotic regulation, acid–base
balance, and the control of extracellular fluid volume (Martinez, 2017).
The excretion system consists of kidneys, ureters, bladder and urethra by
producing urine which carries various metabolic waste products to be disposed of. The
kidneys also function in regulating the body's fluid and electrolyte balance and are the
disposal sites for rennin and erythropitin. Renin plays a role in regulating blood
pressure and erythropitin plays a role in stimulating the production of red blood cells.
Urine is also produced by the kidneys running through the ureter to the bladder through
the urethra (Juncquiera, 1997).
The kidneys are important organs that perform various functions to keep blood
clean and chemically balanced. The kidneys are composed of the skin of the kidneys
(cortex), kidney marrow (medulla), and kidney cavity (pelvis). The kidneys are shaped
like red bean seeds. It is about 10 cm long, weighs approximately 170 grams, and is
located in the abdominal cavity. The kidneys are 2 pieces and are purplish red. The left
kidney is higher than the right kidney. Nephrons are found in the skin of the kidneys
and function as a blood filter. The cortex contains approximately one million nephrons.
Each nephron is composed of a malphighi body and a long (tubular) winding channel.
The malpighi body is composed of the glomerulus and Bowman's capsule. Glomerulus
is a strand of capillary blood where blood is filtered. Glomerulus is surrounded by
capsules of Bowman (Poedjiadi, 2009). The nephron is the functional unit of kidney
and greatly varies in its structure amongst different vertebrates; also the formation of
nephrons shows a variable degree of differences among species. In birds, the kidney
has two kinds of nephrons; one is reptilian type and small sized, without loops of
Henle, and other is mammalian type large in size with long or intermediate length
loops. In contrast, the morphological process of podocytes development has been
reported in the classic kidney of human, horse, goat, monkey, rabbit, cat, dog, chicken,
medaka fish, rat and guinea pig indicating that, it would be a good model system for
studying renal regeneration (Maurya et al., 2018).
The kidneys play a role important in removing substances toxic or poison,
maintain fluid and other substances balance inside the body. The kidneys emit
metabolic waste products from urea, creatinine and ammonia (Aditya et al., 2018). The
kidneys serve essential functions, such as filtration and excretion of metabolic waste
products from the bloodstream, regulation of necessary electrolytes and stimulation of
red blood cell production. They also serve to regulate blood pressure by the use of a
renin-angiotensin-aldosterone system, controlling reabsorption of water, maintaining
the correct pH level as well as chemical balance and intravascular fluid status of the
body. The kidneys also reabsorb glucose and amino acids which may have involved in
regulation of hormonal functions via erythropoietin, calcitriol, and vitamin D
activation (Maurya et al., 2018).

B. Purpose

The objective of this laboratory activity is to analyze the compounds that can
pass through the filter as a illustration of mammalian kidney filtration function.
 II. MATERIAL AND METHODS

A. Material

The materials that used in this practice are biuret solutions, benedict solutions,
lugol solutions, 1% of protein solutions, 1% of amylum solutions, 1% of glucose
solutions, aquadest, and Sartorius filter paper.
The tools that used in this practice are reaction tube, erlenmeyer tube, syringe,
and funnel.

B. Methods

The method used in this practical activity are :

1. 1 mL of solution (protein, glucose, starch and aquadest) is added to the 4 reaction
tubes that were prepared.
2. Each tube is labeled according to the contents of the test solution.
3. 1 mL Biuret solution is added to the tube containing a protein solution and
aquadest.
4. 1 mL of Benedict's solution is added to the test tube containing glucose. The test
tube is placed into boiling water (100oC) for 5 minutes then shaken, observed
and the alteration is written.
5. One drop of lugol solution is added to the test tube containing amylum, observed
and alteration is written.
6. 2 mL test solutions (protein, glucose, amylum, and aquadest) is filtered with filter
paper into 4 reaction tube that were prepared.
7. Steps 2-5 is repeated. The alteration is observed and written.
8. The result is put on the table.
 III. RESULT AND DISCUSSION

A. Result

Table 3.1. Filtration Test Data Using Filter Paper

The intensity of the color The intensity of color
Solution
(before filtration) (after filtration)
Protein +++ ++
Glucose ++ +++
Amylum +++ -
Aquadest + +++

Details:
- = No changing
+ = Low color changing
++ = Medium color changing
+++ = Strong color changing

Figure 3.1. Amylum filtration Figure 3.2. Protein Filtration

Test Test
B. Discussion

Based on the practical activity that have done with glucose solution, starch,
protein, and aquadest, the results were obtained in glucose solution (control) before
filtration that dropped by benedict’s solution is give interpretation that is medium color
changing (++), while after filtration (treated) and dropped by benedict’s solution the
intensity of the color changing was strong (+++). In protein solution (control) before
filtration and dropped by biuret is give interpretation that is strong color changing
(+++) while after filtration (given treatment) and dropped by biuret give interpretation
with medium color changing (++). In amylum or starch solution before filtration
(control) that dropped by KI solution is give interpretation that is strong color changing
(+++), while after filtration and dropped by KI solution the intensity of the color does
not change (-). In aquadest before filtration (control) shows low color changing (+)
while after filtration (treatment) the intensity of the color becomes strong (+++). Some
color intensity before filtration (control) is stronger and weaker than after filtration or
after treatment, because after filtration many substances are filtered, so that the
solution concentration will decrease and will affect the color intensity.
According to Sherwood (2001), states that small materials that can be dissolved
in plasma, such as glucose, amino acids, sodium, potassium, chloride, bicarbonate,
other salts and urea pass through the filter and become part of the sediment. Glucose,
amino acids, and starch are included in substances that will be filtrated and will cause
differences in color intensity, except for protein results that are different from those of
Despopoulus (1998), which states that compounds or large molecules, such as proteins
cannot be filtered by kidney. The color intensity for glucose before and after filtration
remains the same, there is no change in color. The intensity is strong (brick red). This
shows that the glucose solution can pass through the kidney filter. The results are in
accordance with the statement of Guyton (1996), which states that generally molecules
with a 4 nm or more raidus cannot be filtered, otherwise a 2 nm or less molecule will
be filtered indefinitely, small materials that can be dissolved in plasma, such as
glucose, acid amino, sodium, potassium, chloride, bicarbonate, other salts, and urea
will pass through the filter and become part of the sediment.
The mammalian kidney that is meta-nephros, comprises complex epithelial
tubules, nephrons that filter the blood to remove waste and reabsorb water and
nutrients to maintain homeostasis. The human kidney contains up to 2 million
nephrons per organ (Takasato & Little, 2015). The paired kidneys are the central
organs of homeostasis for our body systems. Around 180 L of blood is filtered per day
by the kidneys, accounting for around 20% of cardiac output. Filtering removes
metabolic waste products, and kidney action adjusts water, salt, and pH to maintain
the homeostatic balance of tissue fluids. The kidneys also regulate blood pressure
through the renin–angiotensin–aldosterone system, erythrocyte production through
production of erythropoietin, and circulating calcium and phosphate levels, in part
through the activation of vitamin D (Hoenig & Zeidel, 2014). The kidney is a vital
organ of the mammalian body and becomes a foremost subject of medical research
because many renal diseases in humans are incurable when the kidney is severely
damaged (Maurya et al., 2018).
Urine formation occurs through following three processes there are filtration,
reabsorption, and secretion. During filtration, blood enters the afferent arteriole and
flows into the glomerulus where filterable blood components, such as water and
nitrogenous waste, moves towards the inside of the glomerulus, and non-filterable
components, such as cells and serum albumins, exit via the efferent arteriole. These
filterable components accumulate in the glomerulus to form the glomerular filtrate. On
average, about 20% of the total blood pumped by the heart per minute enters into the
kidneys to undergo filtration. The remaining 80% of the blood flows throughout the
body to facilitate tissue perfusion and proper exchange of gas. During reabsorption,
molecules and ions present in the blood will be reabsorbed into the circulatory system.
The fluid passes through the components of the nephron (i.e. proximal/distal
convoluted tubules, the loop of Henle and collecting duct) as water and ions are
removed due to the change in fluid osmolarity (ion concentration). In the collecting
tube, secretion will occur before the fluid leaves the ureter in the form of urine. During
secretion, some substances such as hydrogen ions, creatinine, and drugs will be
removed from the blood through the peritubular capillary network into the collecting
duct. The end product of all these processes is urine, which is mostly a collection of
substances that have not been reabsorbed during glomerular filtration or tubular
reabsorption process (Maurya et al., 2018).
The content of compounds in urine can be tested using various reagents, one
of which is Biuret as a test material for protein in urine, this is based on reagent Biuret
solution is a blue solution which will be changed to appear violet when exposed contact
with proteins, or other substances that have peptide bonds. In tests carried out the
reagents did not actually contain Biuret, they were so named because both the Biuret
solution and the protein had the same response to the filtration testing process carried
out (Davey, 2005). The principle of Benedict's work on glucose in urine will reduce
kuprisulfate (in benedict) to kuprosulfate which is seen by the color change from
Benedict's solution. When the benedict reagent is mixed and heated with glucose,
where glucose has electrons to give, copper (one of the ingredients in the benedict
reagent) will receive the electron and undergo reduction so that color changes occur.
So, if urine contains glucose, a color change reaction will occur as described above.
However, if there is no glucose, then the reaction will not occur and the color of
benedict will not change (Carlton & Mc Gavin, 1995).
The function of the biuret reagent is to form a complex so that what is contained
can be identified. This biuret reaction is specific, meaning that only compounds
containing pepetida bonds will react with biuret reagents. Benedict is a reagent to test
the content of foods containing glucose. Just like testing using biuret, the food
ingredients tested must be in the form of a solution, then added benedict reagents
(usually half of the amount of solution). After that it is heated for a few minutes. Food
containing glucose, there will be green to brick red deposits, green if the glucose
content is small and brick red if there is a lot of glucose content. In addition to using
benedict, glucose testing can also use the FehlingA + FehlingB reagent. Lugol reagents
indicate that a substrate contains starch and will produce a blue black color (Machin,
2012).
In the process of filtration until urinary excretion in the kidneys can be
influenced by factors that can affect the amount or state of urine, namely, the amount
of water taken, the state of the nervous system, ADH hormone, the amount of salt that
must be removed from the blood so that the pressure becomes osmotic, in diabetics
Glucose mellitus followed by increased urine volume. ADH functions as a hormone
that facilitates the absorption of water from the distal tubule to collecting ducts. These
ADH hormones influence each other with the concentration of water in the body. If
the water concentration decreases, ADH will flow with the blood which results in
increased vascular permeability and absorbed water. Less urine is formed. Conversely
if the concentration of high water in the blood, ADH secretion will decrease and cause
absorption of water in the distal vessels decreases. Urine becomes more numerous and
aqueous. This urine formation process while balancing fluid levels in the body that are
not needed. If the consumption of water in large quantities, then the urine produced
will also be a lot and the voiding process becomes more frequent. Increased
concentration of water in the blood reduces colloid pressure and pressure during
fitration. This resulted in reduced absorbed water and increased urine production. The
amount of water in the blood, makes the colloid pressure smaller so that the absorption
process does not run optimally and the water loss is immediately carried out. Lack of
insulin levels in the body will increase glucose levels for example in patients with
diabetes mellitus. This high sugar level will interfere with the process of reabsorption
in the distal tubule. High glucose makes blood flow or blood viscosity more thick so
it is more difficult to absorb. This can also trigger damage to the kidneys if it occurs
continuously. Kidney burden gets heavier with more viscous blood viscosity
(Thenawijaya & Maria, 1995).
Kidney disease is also defined as a disorder that affects the kidneys which
results from various factors, such as infections, tumors, congenital abnormalities,
metabolic or degenerative diseases, and others. Chronic kidney failure is a progressive
disorder of kidney function and irreversibIe. When the kidney is unable to carry out
its function it will cause kidney failure. This process ultimately results in decreased
urine production and kidney failure, with buildup of waste products in the blood and
body tissues. One common reason for kidney failure in the United States is diabetes.
Sometimes chronic kidney disease is accompanied by high blood pressure, which not
only can be caused by kidney damage but also further accelerates kidney injury and is
a major reason for the negative effects of chronic kidney disease on other organs,
including increased risk of heart disease and stroke, collection of excess body fluids,
anemia, weakening of bones, and impairment of the way the body eliminates
medications (Razmaria, 2016).
Diabetes is the main cause of kidney disease. Around 1 of 4 adults with
diabetes have kidney disease. Kidney damage due to chronic diabetes mellitus is often
found. Kidney hypertrophy due to increased work must be done by the kidneys of
chronic diabetics to reabsorb glucose. High blood glucose levels make the kidneys
filter too much blood. High blood glucose can damage blood vessels in kidneys. When
blood vessels are damaged, they don't work properly. Many people with diabetes also
experience high blood pressure, which can damage the kidneys. Diabetes mellitus
(DM) or abbreviated as Diabetes is a health disorder in the form of a collection of
symptoms caused by an increase in blood sugar (glucose) levels due to lack or insulin
resistance. DM is a disease where a person secretes a large amount of urine that feels
sweet. There are at least three forms of diabetes mellitus, namely type I, type II, and
gestational diabetes (Aditya et al., 2018). Albuminuria is a disease that is shown by
the presence of albumin and other proteins in the urine so the kidneys cannot carry out
the screening process, especially protein screening. Because protein (albumin) is not
filtered, the protein can come out with urine. Albuminuria is the cause because of
damage to the filtration device. Increased levels of lactate can also be found in patients
with Chronic Kidney Disease (CKD) and one way to overcome this is with HD
(Hemodyalysis) (Shima et al., 2011).
Kidney stones or nephrolithiasis can form due to the deposition of calcium
salts in the kidney cavity, renal tract or bladder. Kidney stones are crystals that are
insoluble and contain calcium oxalate, uric acid, and calcium phosphate crystals. The
reason is because you consume too much mineral salt and consume too little water.
These kidney stones can further cause hydronephrosis. Hydronephrosis is the
enlargement of one kidney because urine cannot flow out. This is due to narrowing of
the flow of the kidneys or blocked by kidney stones. Glucosuria is a disease
characterized by glucose in the urine. The disease is often also called diabetes or
diabetes (diabetes mellitus). Glucose levels in the blood increase due to a lack of
insulin. Nephrons are not able to reabsorb excess glucose, so excess glucose is
removed with urine. Nephritis is damage to the glomerular part of the kidneys due to
allergic germ poisons. Nephritis is usually caused by the presence of Streptococcus
bacteria. Hematuria is a disease characterized by the presence of red blood cells in the
urine. The disease is caused by inflammation of the urinary organs or due to irritation
due to friction in kidney stones. (Wilson, 1979).
 IV. CONCLUSION

Based on the result can be concluded that the compounds filtered in the filtration
process are protein, glucose and starch. Aquadest cannot be filtered and remain past
the kidneys.
 REFERENCES

Aditya, A., Udiyono, A., Saraswati, L. D. & Setyawan, H., 2018. Screening Fungsi
Ginjal sebagai Perbaikan Outcome Pengobatan pada Penderita Diabetes
Mellitus Tipe II (Studi di Wilayah Kerja Puskesmas Ngesrep). Jurnal
Kesehatan Masyarakat, 6(3), pp. 191-199.

Carlton, W.W. & Mc Gavin, M. D., 1995. Special Veterinary Pathology. United State
of America: Mosby.
Dahelmi., 1991. Fisiologi Hewan. Padang: UNAD press.
Davey, P., 2005. At A Glance Medicine. Jakarta: Erlangga.
Despopoulus., 1998. Biokimia. Jakarta: Erlangga.
Guyton, M., 1996. Buku Ajar Ilmu Penyakit Dalam. Jakarta: Interna Publishing.

Hoenig, M. P. & Zeidel, M. L., 2014. Homeostasis, the Milieu Inte´rieur, and the
Wisdom of the Nephron. Clinical Journal of the American Society of
Nephrology, 9, pp.1-10.

Juncquiera, L. C., 1997. Histologi Dasar. Jakarta: Penerbit Buku Kedokteran.

Machin, A., 2012. Potensi Hidrolisat Tempe Sebagai Penyedap Rasa Melalui
Pemanfaatan Ekstrak Buah Nanas. Jurnal Biosaintifika, 4(2), Pp: 75-91.

Maurya, H., Kumar, T. & Kumar, S., 2018. Anatomical and Physiological Similarities
of Kidney in Different Experimental Animals Used for Basic Studies. Journal
of Clinical & Experimental Nephrology, 3(2), pp. 1-6.

Martinez, C. B., 2017. The Kidney. Brazil: State University of Londrina.

Poedjiadi, A., 2009. Dasar-Dasar Biokimia. Jakarta: Universitas Indonesia.

Razmaria, A. A., 2016. Chronic Kidney Disease. JAMA journal, 315(20), pp. 1-3.

Shima T. S., Khatun, A., Yeasmin F., Ferdousi, S., Kirtania, K. & Sultana N. 2011.
Cystatin C: A Better Predictor of Kidney Function in Diabetic Patients.
Bangladesh. J Med Biochem, 4(1), pp. 16-20.

Sherwood, L., 2001. Fisiologi Manusia daru Sel ke Sistem. Jakarta: Penerbit Buku
Kedokteran EGC.

Takasato, M. & Little, M. H., 2015. The Origin of the Mammalian Kidney:
Implications for Recreating the Kidney in Vitro. Development Journal, 142,
pp. 1937-1947.

Thenawijaya & Maria C., 1995. Biokimia Nutrisi dan Metabolisme. Jakarta : UI Press.
Wilson, J. A., 1979. Prinsiple of Animal Physiology. London: Collier Mc Millan
Publisher.