Comparing the Effect of Methods of Rat Euthanasia on the Brain of Wistar Rats: Cervical Dislocation, Chloroform Inhalation, Diethyl Ether Inhalation and Formalin Inhalation

2020 ◽  
pp. 8-16
Author(s):  
U. S. Aguwa ◽  
C. E. Eze ◽  
B. N. Obinwa ◽  
S. N. Okeke ◽  
S. F. Onwuelingo ◽  
...  
Keyword(s):  
Diethyl Ether ◽  
Wistar Rats ◽  
Phosphate Buffer Solution ◽  
Control Group ◽  
Buffer Solution ◽  
Histological Studies ◽  
Male Wistar Rats ◽  
Antioxidant Parameters ◽  
Cervical Dislocation ◽  
The Brain

Contentions still exist as to the most humane method of sacrificing experimental animals. Beyond humaneness, there is also the issue of appropriateness of a method of sacrifice with regards to organ safety. This study compared four common methods of rodent sacrifice used in Nigeria and their effects on the brain. Twenty male Wistar rats weighing 150 to 180g were randomly separated into four groups of five rats each. After a two week period of acclimatization, the animals were sacrificed; Group A by Cervical dislocation (CD) which also served as control, Group B by chloroform inhalation (CI), Group C by diethyl ether inhalation (DEI) and group D by Formalin inhalation (FI).  The time taken for the animals to pass out was documented and the brains were harvested. Four of the brains from rats in each group were homogenized in phosphate buffer solution and centrifuged at 10000rpm. The supernatant were used for antioxidant studies. The remaining one brain from each group were fixed in 10% formal saline and after 48 hours, the cerebellum (CB) and hippocampus (Hp) were used for histological studies using the H & E method. Our results show although CD was the quickest, it gave the least quantity of blood. Meanwhile CI was the most humane, gave the most blood and saved time compared to DEI and FI. None of the methods of sacrifice showed adverse effect on the antioxidant parameters of the rats. However, histological studies showed that while CD and CI showed no adverse effects on the hippocampus, only CD was completely safe for the CB. The other methods showed various levels of cell death. It is therefore expedient to consider these factors in making the choice of an appropriate method of sacrifice and to use the most humane and safest method with reference to the organ studied.

scholarly journals Evaluating the Effect of Chloroform Inhalation as a Method of Euthanasia on the Cerebellum and Hippocampus of Adult Wistar Rats

2020 ◽  
pp. 14-25
Author(s):  
Ugochukwu Samuel Aguwa ◽  
Okeke Somadina Nnamdi ◽  
Ezejindu Darmian Nnabuihe ◽  
Eze Chinyere Elizabeth ◽  
Azurunwa Ogechi ◽  
...  
Keyword(s):  
Wistar Rats ◽  
Dose Dependence ◽  
Dose Dependent Fashion ◽  
Male Wistar Rats ◽  
Antioxidant Parameters ◽  
Group A ◽  
Cervical Dislocation ◽  
Group B ◽  
Dose Dependent ◽  
The Brain

Chloroform inhalation is a common method of rodent euthanasia in Nigeria for research purposes. This work is designed to evaluate the consequence of this method of sacrifice on the cerebellum and hippocampus of Wistar rats. Twenty male Wistar rats weighing between 160 and 180 were divided into 4 groups of 5 rats each. Group A served as control and was sacrificed using cervical dislocation (a widely acceptable non-inhalation method of sacrifice). Groups B, C and D were exposed to chloroform for 5 minutes once a day (group B), once a day for 2 days and once a day for 3 days. The brains were removed; four from each group was processed for antioxidant assay while one from each group was fixed in Bouin’s fluid for histological studies. Our results show that chloroform inhalation adversely affected the results of the antioxidant parameters studied in a dose-dependent fashion. That means that the adverse effect worsened as the number of days increased. This was also the case with the histology results as there was evidence of cell necrosis in the cerebellar and hippocampal tissues. This also showed dose dependence. We therefore conclude from our results that when studying the brain tissues or carrying out brain related researches, chloroform inhalation is not the method of choice for rat euthanasia.

scholarly journals Impact of Vanadium Complexes Treatment on the Oxidative Stress Factors in Wistar Rats Plasma

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
R. Francik ◽  
M. Krośniak ◽  
M. Barlik ◽  
A. Kudła ◽  
R. Gryboś ◽  
...  
Keyword(s):  
Wistar Rats ◽  
Reduced Glutathione ◽  
The Body ◽  
Stress Factors ◽  
Control Group ◽  
Vanadium Complexes ◽  
Healthy Male ◽  
Reducing Ability ◽  
Male Wistar Rats ◽  
Antioxidant Parameters

The aim of this study was to investigate the clinical efficacy of vanadium complexes on triglycerides (TG), total cholesterol (Chol), uric acid (UA), urea (U), and antioxidant parameters: nonenzymatic (FRAP—ferric reducing ability of plasma, and reduced glutathione—GSH) and enzymatic (glutathione peroxidase—GPx, catalase—CAT, and GPx/CAT ratio) activity in the plasma of healthy male Wistar rats. Three vanadium complexes: [VO(bpy)2]SO4⋅2H2O, [VO(4,4′Me2bpy)2]SO4⋅2H2O, and Na[VO(O2)2(bpy)]⋅8H2O are administered by gavage during 5 weeks in two different diets such as control (C) and high fatty (F) diets. Changes of biochemical and antioxidants parameters are measured in plasma. All three vanadium complexes statistically decrease the body mass growth in comparison to the control and fatty diet. In plasma GSH was statistically increased in all vanadium complexes-treated rats from control and fatty group in comparison to only control group. Calculated GPX/CAT ratio was the highest in the control group in comparison to others.

Oophorectomy hinders antioxidant adaptation promoted by swimming in Wistar rats

2013 ◽  
Vol 38 (2) ◽  
pp. 148-153 ◽  
Author(s):  
Ulisvaldo Brunno de Oliveira Macedo ◽  
Rand Randall Martins ◽  
Francisco Paulo Freire Neto ◽  
Yonara Monique da Costa Oliveira ◽  
Aldo da Cunha Medeiros ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Glutathione Peroxidase ◽  
Wistar Rats ◽  
Antioxidant Defense ◽  
Hormone Replacement ◽  
Antioxidant Defense System ◽  
Control Group ◽  
Defense System ◽  
Antioxidant Parameters ◽  
The Brain

Oxidative stress is associated with postmenopause and is also responsible for various metabolic alterations. The redox imbalance observed during ovarian decline can be induced experimentally by bilateral ovariectomy in rats. In addition to hormone replacement, regular moderate physical exercise is indicated to prevent several common postmenopausal diseases. This study aimed to assess the effect of daily swimming on the antioxidant defense system of oophorectomized Wistar rats. Control and oophorectomized groups were submitted to 1 h of daily swimming for 90 days. Levels of lipid peroxidation and glutathione content and the activities of superoxide dismutase enzyme and glutathione peroxidase in erythrocytes, liver, and brain were assessed every 30 days. The control group exhibited lower lipoperoxidation that was associated with a significant increase in superoxide dismutase enzyme activity, glutathione peroxidase activity, and glutathione content in erythrocytes and liver; however, swimming did not cause changes in antioxidant parameters in the brain over time. The oophorectomized group showed no antioxidant adaptation to daily swimming and had greater oxidative damage in the liver and blood. Our results suggest that ovariectomy hinders antioxidant adaptation in Wistar rats submitted to daily swimming.

scholarly journals Effect of Artesunate Treatment on Some Brain Biomolecules and its Behavioral Implication

1970 ◽  
Vol 4 (2) ◽  
pp. 44-50
Author(s):  
MB Ekong ◽  
AO Igri ◽  
TB Ekanem
Keyword(s):  
Therapeutic Dose ◽  
Wistar Rats ◽  
Tap Water ◽  
Control Group ◽  
Anthropometric Parameters ◽  
Whole Brain ◽  
Long Duration ◽  
Male Wistar Rats ◽  
Group A ◽  
The Brain

Background: Artesunate (AS) is an artemisinin antimalarial drug used as a single drug or in combination with other antimalarials. Objective: This study was to find its effect on some brain biomolecules and behavioural activities in Wistar rats. Methods: Forty adult male Wistar rats weighing between 150- 180g were divided into four groups of A, B, C and D with 10 animals each. Group A served as the control that received tap water, while groups B, C and D served as the experimental groups that received 2.85mg/kg (therapeutic dose-TD) and 5.71mg/kg (high pharmacologic dose-HPD) of AS per day for 3 days, and 2.85mg/kg (long duration therapeutic dose -LDTD) of AS per day for six days respectively. Half of the dose was administered twelve hourly (twice a day), and twelve hours after the last treatments, behaviour test using the ‘open field maze' was carried out. Immediately after, the animals were sacrificed with chloroform anaesthesia and the whole brain removed and weighed. Whole brain homogenates were used to determine brain total protein (TP), triacylglycerol (TAG) and cholesterol (CH).Data were analyzed statistically by ANOVA and Tukey-Kramer Multiple Comparative Test as applicable. Results: There were no difference (p<0.05) between the experimental groups and the control group in the anthropometric parameters and behavioural activities. In the brain biomolecules concentration, TP was lower in concentration in the HPD group, TAG was lower in concentration in the LDTD group, while the HPD and LDTD groups had lower CH concentration compared to the control. In all the parameters studied no difference was found between the TD group and the control. Conclusion: AS at recommended dose may not affect some behaviour and brain biomolecule concentration, unlike when taken in excess of dose and or time. Even at these doses/time there may have been no behavioural manifestation. Key words: Artesunate, Brain, Anthropometry, Behavior, Biomolecules, Rats DOI: 10.3329/jbsp.v4i2.4171 J Bangladesh Soc Physiol. 2009 Dec;4(2): 44-50  

scholarly journals Monosodium Glutamate Maintains Antioxidant Balance in the Neuro-Retinal Axis of Male Wistar Rats

2021 ◽  
Author(s):  
Halima Nalugo ◽  
Herbert Izo Ninsiima ◽  
Keneth Iceland Kasozi ◽  
Ritah Nabirumbi ◽  
Lawrence Obado Osuwat ◽  
...  
Keyword(s):  
Catalase Activity ◽  
Wistar Rats ◽  
Structural Changes ◽  
Monosodium Glutamate ◽  
Food Additives ◽  
Control Group ◽  
Human Consumption ◽  
Group I ◽  
Male Wistar Rats ◽  
The Brain

Abstract Purpose Information on monosodium glutamate (MSG) safety in the eye-brain axis remains scarce. The objective of the study was to establish changes in calcium (Ca2+) concentrations, catalase activity, Malondialdehyde (MDA) levels and identify any major histological changes associated with parenteral and feed supplementation of MSG in male Wistar rats. This is important to guide policy on human consumption of food additives.Methods Thirty adult male Wistar rats were divided into 5 groups (n = 6) for a period of one month. Group I and IV were treated with MSG 6g/kg and 60 mg/kg subcutaneous, while groups II and III at 5% and 2% MSG feed supplement. Group V was the control group treated with normal saline (no MSG). The eye and brain were analyzed for Ca2+, catalase, MDA levels and histological analysis was done using Hematoxylin and Eosin staining.Results Calcium levels in the eye was reduced at high (6g/kg/day and 5% s.c) MSG treatments while in the brain these remained constant. Since Ca2+ plays a crucial role in catalase activity, levels of catalase were not affected in eye-brain axis. MDA levels were elevated at high MSG treatments and no structural changes in the retina of the eye and no damage were found in the brain.Conclusion MSG maintained catalase levels, Ca2+ and no structural changes were observed in the eye and brain.

scholarly journals The effect of Dichlorvous (Sniper) Inhalation on the Histology of the Lungs of Adult Male Wistar Rats

2021 ◽  
pp. 58-67
Author(s):  
A. E. Anyabolu ◽  
D. N. Ezejindu ◽  
B. N. Obinwa
Keyword(s):  
Wistar Rats ◽  
The Body ◽  
Control Group ◽  
Male Wistar Rats ◽  
Water Feed ◽  
Group A ◽  
Cervical Dislocation ◽  
Group B ◽  
Histological Architecture ◽  
Group D

The organophosphate, dichlorvous (Otapia-pia) formulated in varying concentrations as insecticides is utilized by several individuals in most remote places of Nigeria due to its affordable value and accessibility. However, this present study is conducted to investigate the adverse effect of the exposure of this substance on the respiratory system (lungs) of male albino wistar rats. Twenty (20) albino wistar rats comprising of all males weighing between 150-230g were divided into four groups (A-D) of five animals each. Group A received only water, feed, and served as the control. Group B was exposed to dichlorvous inhalation 3hrs daily for a period of two weeks; Group C was exposed to dichlorvous inhalation 6hrs daily for a period of two weeks; while Group D was exposed to dichlorvous inhalation 10hrs daily for a period of two weeks. Twenty four hours after the last exposure, the animals were sacrificed by cervical dislocation and dissected. The lungs were weighed and fixed in 10% formal saline for histological studies. The body weight of the experimental groups decreased insignificantly when compared with the control group. The lungs weight increased significantly when compared to the control groups. Histological observation revealed a moderate to severe effects on the lungs with severe consolidated inflammatory exudates (discharge of fluids from pores), a moderate intra-alveoli hemorrhage, and a severe dilation of alveolar septa which is evident as emphysematous changes. This result revealed that the exposure to dichlorvous causes distortion of the normal histological architecture of the lungs. Thus, showing that dichlorvous inhalation influences the normal physiologic mechanisms of the lungs.

Chronic immobilization stress induces anxiety-related behaviors and affects brain essential minerals in male rats

2020 ◽  
pp. 1-8
Author(s):  
Zafer Sahin ◽  
Alpaslan Ozkurkculer ◽  
Omer Faruk Kalkan ◽  
Ahmet Ozkaya ◽  
Aynur Koc ◽  
...  
Keyword(s):  
Immobilization Stress ◽  
Central Area ◽  
Essential Elements ◽  
Male Rats ◽  
Control Group ◽  
Male Wistar Rats ◽  
Swimming Test ◽  
The Brain ◽  
Center Area ◽  
Chronic Immobilization Stress

Abstract. Alterations of essential elements in the brain are associated with the pathophysiology of many neuropsychiatric disorders. It is known that chronic/overwhelming stress may cause some anxiety and/or depression. We aimed to investigate the effects of two different chronic immobilization stress protocols on anxiety-related behaviors and brain minerals. Adult male Wistar rats were divided into 3 groups as follows ( n = 10/group): control, immobilization stress-1 (45 minutes daily for 7-day) and immobilization stress-2 (45 minutes twice a day for 7-day). Stress-related behaviors were evaluated by open field test and forced swimming test. In the immobilization stress-1 and immobilization stress-2 groups, percentage of time spent in the central area (6.38 ± 0.41% and 6.28 ± 1.03% respectively, p < 0.05) and rearing frequency (2.75 ± 0.41 and 3.85 ± 0.46, p < 0.01 and p < 0.05, respectively) were lower, latency to center area (49.11 ± 5.87 s and 44.92 ± 8.04 s, p < 0.01 and p < 0.01, respectively), were higher than the control group (8.65 ± 0.49%, 5.37 ± 0.44 and 15.3 ± 3.32 s, respectively). In the immobilization stress-1 group, zinc (12.65 ± 0.1 ppm, p < 0.001), magnesium (170.4 ± 1.7 ppm, p < 0.005) and phosphate (2.76 ± 0.1 ppm, p < 0.05) levels were lower than the control group (13.87 ± 0.16 ppm, 179.31 ± 1.87 ppm and 3.11 ± 0.06 ppm, respectively). In the immobilization stress-2 group, magnesium (171.56 ± 1.87 ppm, p < 0.05), phosphate (2.44 ± 0.07 ppm, p < 0.001) levels were lower, and manganese (373.68 ± 5.76 ppb, p < 0.001) and copper (2.79 ± 0.15 ppm, p < 0.05) levels were higher than the control group (179.31 ± 1.87 ppm, 3.11 ± 0.06 ppm, 327.25 ± 8.35 ppb and 2.45 ± 0.05 ppm, respectively). Our results indicated that 7-day chronic immobilization stress increased anxiety-related behaviors in both stress groups. Zinc, magnesium, phosphate, copper and manganese levels were affected in the brain.

Pengaruh Profilaksis Ekstrak Ubi Jalar Ungu (Ipomoea batatas L.) terhadap Kadar Malondialdehida Hepar Tikus Putih (Rattus norvegicus) Jantan Galur Wistar yang Diinduksi Karagenan

2018 ◽  
Vol 15 (2) ◽  
pp. 146
Author(s):  
BRILIAN DINANTI ◽  
FITRI HANDAJANI
Keyword(s):  
Sweet Potato ◽  
Ipomoea Batatas ◽  
Wistar Rats ◽  
Group Method ◽  
Control Group ◽  
Intraplantar Injection ◽  
Significant Difference ◽  
Male Wistar Rats ◽  
Potato Extract ◽  
Purple Sweet Potato

<p>Liver is an organ with complex metabolism. When the liver is inflamed, cellular immunity will defend against inflammatory agents by stimulating immune cells to produce reactive oxygen species (ROS). Excessive ROS accumulation cause oxydative stress with increased  liver malondialdehyde (MDA) level. Some researches showed that purple sweet potato contain flavonoids (anthocyanins) that functioned as antioxydants. This study aimed to show the prophylactic effect of purple sweet potato extract to the liver MDA level of male Wistar rats induced by carrageenan.</p><p>This study used post-only control group method using 18 male Wistar rats divided into 3 groups: group of rats without treatment, group of rats induced by 0,1 ml of 1% carrageenan by intraplantar injection on day-8, and group of rats given with 872 mg/kgBW of purple sweet potato extract for 7 days and induced by 0,1 ml of 1% carrageenan. In the end of the study, the liver MDA levels were measured by Thio-Barbituric Acid method on each groups.</p><p>The results of One-Way ANOVA test showed there was no significant difference (p = 0,290) between group of rats without treatment (<em>x̅</em>= 207,50) and group of rats induced by carrageenan (<em>x̅</em>=233,17). Then, there is no significant difference (p = 0.978) between group of rats induced by carrageenan and group of rats given with prophylactic purple sweet potato extract and induced by carrageenan (<em>x̅</em>= 232,50).</p><p>The conclusion of this study is giving intraplantar injection of carrageenan can increase liver MDA level insignificantly and giving prophylactic purple sweet potato extract has an effect to decrease the liver MDA level of rats induced by carragenan insignificantly because it contains anthocyanins as antioxidants.</p><p> </p><strong>Keywords: </strong>Liver, <em>Ipomoea batatas</em> L., Malondialdehyde, Anthocyanins

scholarly journals Anticarcinogenic effect of probiotic fermented milk and chlorophyllin on aflatoxin-B1-induced liver carcinogenesis in rats

2011 ◽  
Vol 107 (7) ◽  
pp. 1006-1016 ◽  
Author(s):  
M. Kumar ◽  
V. Verma ◽  
R. Nagpal ◽  
A. Kumar ◽  
P. V. Behare ◽  
...  
Keyword(s):  
Wistar Rats ◽  
Lactobacillus Rhamnosus ◽  
Thiobarbituric Acid ◽  
Fermented Milk ◽  
Hepatoprotective Effect ◽  
Control Group ◽  
Time Interval ◽  
Group I ◽  
Male Wistar Rats ◽  
Aflatoxin B

The present investigation was carried out to evaluate the hepatoprotective effect of probiotic fermented milk (FM) containing Lactobacillus rhamnosus GG and Lactobacillus casei strain Shirota, alone as well as in combination with chlorophyllin (CHL) as an antioxidant agent in male Wistar rats administered aflatoxin-B1 (AFB1). AFB1 was injected intraperitoneally at the rate of 450 μg/kg body weight per animal twice a week for 6 weeks, maintaining an equal time interval between the two consecutive AFB1 administrations. A total of 125 male Wistar rats were randomly allocated to five groups, each group having twenty-five animals. Group I was offered FM containing L. rhamnosus GG and L. casei strain Shirota. Group II was administered AFB1 and served as the control group; group III was administered FM-AFB1, in which besides administering AFB1, FM was also offered. Group IV was offered CHL and AFB1, and group V was offered both FM and CHL along with AFB1. The rats were euthanised at the 15th and 25th week of the experiment and examined for the biochemical and hepatopathological profile. A significant reduction in thiobarbituric acid-reactive substances (TBARS) was observed in the FM–CHL–AFB1 group compared with the AFB1 control group. FM alone or in combination with CHL was found to show a significant (P < 0·05) hepatoprotective effect by lowering the levels of TBARS and by enhancing the activities of antioxidant enzymes such as glutathione peroxidase, superoxide dismutase, catalase and glutathione-S-transferase, indicating that probiotic FM alone or in combination with CHL possesses a potent protective effect against AFB1-induced hepatic damage.

IMMUNOHISTOCHEMICAL STUDIES OF BLOOD BRAIN BARRIER AFTER ADMINISTRATION OF ABHRAK BHASM IN WISTAR RATS

2021 ◽  
pp. 13-19
Author(s):  
Amita Singh ◽  
Raj Kumar ◽  
S. K. Kannaujia ◽  
Manikrishna Manikrishna ◽  
N. P. Singh
Keyword(s):  
Blood Brain Barrier ◽  
Wistar Rats ◽  
Brain Parenchyma ◽  
Brain Barrier ◽  
Major Constituent ◽  
Control Group ◽  
Blood Brain ◽  
Biological Studies ◽  
Immunohistochemical Studies ◽  
The Brain

Abhrak bhasma (AB) is a type of bhasma prepared from repeated incineration of mineral mica with decoctions of about 72 herbs. The particle size of Abhrak bhasm has been shown to be in the range of 29-88 nanometers and Fe, Ca, Si, Mg and K are found to be as major constituent. Many drugs developed to treat Central Nervous System (CNS) disorders are unable to reach the brain parenchyma in therapeutically relevant concentrations. The blood brain barrier protects brain parenchyma from the uctuation of plasma composition, from pathogenic agents and maintains homeostasis of the brain parenchyma by restricting non-specic ux of ions, peptides, proteins and even cells into and out the brain. Immunohistochemistry is being widely employed as a tool for biological studies. This study is conducted to examine the change in the continuity of Blood brain barrier by using immunohistochemistry, once Abhrak bhasm drug is given in experimental animal and also to examine the histology of organs. In this study a total of 30 adult albino Wistar rats of approximately 4 months age (approx. 150-200 gms) of either sex selected randomly to see the effect of Abhrak bhasm, an ayurvedic drug on Wistar rats. The rats were weighed, marked and divided into 5 groups each consisting of six animals. In normal control group (Group E), no drug was administered and in rest of the four treated groups (Group-A,B,C,D), Abhrak bhasm @ 36 mg/kg B.wt. was administered orally once in each rat. Brain, liver, kidneys,spleen and blood samples were collected in 10% formalin solution after euthanizing the rats at 0.5,2,6 & 12 hours of Abhrak bhasma drug intervention. The alterations in any of the biochemical parameters are within the tolerable limits of liver and kidney since the dose of abhrak bhasm did not affect liver and kidneys. In the present study, the increase in ALP level may be the result of alterations in metabolisms that occurred without any signicant alteration in histology of liver. After applying the immunohistochemistry with the research markers GFAP, CD 34, S 100, GLUT-1 and RECA-1 on the rats in groups A,B,C and D, there was no change in the intensity of immunohistochemistry, with respect to control. While on applying the Occludin, the intensity of immunohistochemistry was reduced in all the treatment groups as compared to the control group. On the basis of ndings of present study it can be concluded that the therapeutic dose of Abhrak bhasma causes changes at the level of tight junctions present in blood brain barrier in rats which is shown by immunohistochemistry with occludin research marker. There is no toxic effect of drug on different organs of rats as no signicant changes in histology of organs are seen. More studies need to be done to check the permeability of blood brain barrier for Abhrak bhasma drug, like calculating its concentration in brain tissues and other vital organs of rat.

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