a-collection-of-research-articles-on-the-medical-potential-of-cow-urine

790 E.E. Jarald et al.
Downloaded by [University <strong>of</strong> Gothenburg] at 21:45 17 September 2015
Materials and Methods
Procurement <strong>of</strong> materials
The urine <strong>of</strong> a 2-year-old virgin Gujarati Indian cow known
as “Geer cow” was used in the study. The study was performed
after getting a certificate from the veterinary doctor
stating that the cow was free from diseases. Fresh cow urine
was collected daily and used after filtration. The plant drugs
were collected from the Gayathri Parivar (local traditional
healers) in order to minimize the variation in the claimed
therapeutic effect. The collected plant materials were positively
identified by Dr. H.S. Chatree, Botanist, Govt. Arts
and Science College, Mandsaur, and the voucher specimens
were retained in our department for future reference.
Preparation <strong>of</strong> extracts
The herbal preparations using cow urine and distilled water
were made using the above-mentioned different plant
species. Equal quantities <strong>of</strong> air-dried samples <strong>of</strong> each plant
species were ground and mixed with 10-times the equivalent
volume <strong>of</strong> cow urine and water separately and boiled
for 4 h. The extracts were filtered and evaporated in a distillation
assembly to get the residue. The percentage yield <strong>of</strong>
extracts prepared using cow urine and distilled water was
12.5% and 11.0%, w/w, respectively. Preliminary chemical
investigation was carried out in the extracts to identify
the nature <strong>of</strong> constituents present in the extracts (Brain &
Turner, 1975; Khandelwal, 2005).
Animals and treatment
After getting approval from the institutional animal ethical
committee (reg. no. – 918/ac/05/CPCSEA), male Wistar
strain rats (weighing between 150 and 200 g) procured
from the animal house <strong>of</strong> B. R. Nahata College <strong>of</strong> Pharmacy,
Mandsaur, were used for the investigation. The animals
were housed in standard environmental conditions <strong>of</strong>
temperature (21 ± 2 ◦ C), humidity (55 ± 10%), and a 12-h
light-dark cycle. Rats were supplied with standard pellet
diet and water ad libitum.
Acute toxicity studies
The acute toxicity test <strong>of</strong> the preparations and cow urine was
determined according to the OECD guidelines (No. 420,
Organization for Economic Cooperation and Development).
Female albino mice (20–25 g) were used for this
study. Dosing amounts for sample in liquid form were calculated
with the help <strong>of</strong> density or specific gravity. After
the sighting study, a starting dose <strong>of</strong> 2000 mg/kg (p.o.) <strong>of</strong>
the test samples was given to various groups <strong>of</strong> five animals
each. The treated animals were monitored for 14 days for
mortality and general behavior. No deaths were observed
through the end <strong>of</strong> the study. The test samples were found
to be safe up to the dose <strong>of</strong> 2000 mg/kg, and doses <strong>of</strong> 200
and 400 mg/kg were chosen for further experimentation.
Antihyperglycemic activity
Diabetes was induced in rats by injecting 150 mg/kg
<strong>of</strong> alloxan monohydrate intraperitoneally in 0.9% w/v
NaCl (Ainapure et al., 1985; Porchezian et al., 2000).
Seventy-two hours after injection, blood glucose level was
measured, and the diabetic rats were divided into eight
groups <strong>of</strong> six animals each. Insulin [1 unit/kg (i.p.)] was
used as standard drug (Mukherjee, 2002). The first group
was kept as vehicle control, the second was treated with
insulin, and the third to eighth groups were treated with
herbal preparations prepared using cow urine, distilled
water, and pure cow urine at two dose levels, 200 and 400
mg/kg (p.o), respectively. One more group was included
in the study to determine the effects <strong>of</strong> fresh cow urine in
the blood glucose level <strong>of</strong> normal rats. Fresh cow urine at
a dose <strong>of</strong> 400 mg/kg was given to the rats in this group for
21 days. The treatment was given once daily for 21 days.
Blood samples were collected at regular intervals after
fasting overnight, before treatment, from rat-tail vein under
mild anesthesia and monitored. The blood sugar level was
monitored using Accu-chek Active Test strips in Accu-chek
Active Test meter (Roche Diagnostics, Germany).
Statistical analysis
Data were expressed as mean ± SEM, and the obtained data
were subjected to one-way ANOVA followed by Dunnet’s
test. The p values less than 0.05 were considered as significant.
Results
The phytochemical investigations performed in the extracts
revealed the presence <strong>of</strong> alkaloids, tannins, flavonoids, carbohydrates,
and saponins in both the extracts. The results
<strong>of</strong> antidiabetic activity <strong>of</strong> cow urine and herbal preparations
prepared using cow urine and water are presented in Table 1.
The basal blood glucose levels <strong>of</strong> all the groups were
statistically not different from each other. Three days after
alloxan administration, blood glucose values were 5-fold
higher in all the groups and were not statistically different
from each other. After 21 days, values <strong>of</strong> blood glucose were
decreased in all the treatment groups (P < 0.05, P < 0.01).
The value in diabetic control group remained stable. The
preparations exhibited activity in a dose-dependent manner.
The activities <strong>of</strong> the preparations were found significant
from the 7th day onwards, whereas the activity <strong>of</strong> cow urine
was found significant only after 21 days <strong>of</strong> treatment. Normal
rats treated with cow urine for 21 days did not show any
elevation in their blood glucose levels. Comparatively, the
preparations containing cow urine were found to be better
than the herbal preparation prepared using distilled water.