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

Author(s):  
Ugochukwu Samuel Aguwa ◽  
Okeke Somadina Nnamdi ◽  
Ezejindu Darmian Nnabuihe ◽  
Eze Chinyere Elizabeth ◽  
Azurunwa Ogechi ◽  
...  

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.

Author(s):  
A. J. Ajibade ◽  
A. E. Okeleye ◽  
I. A. Ogunmola

Cobalt induces hypoxia in the brain which leads to oxygen deprivation resulting in cognitive disturbance and decreased motor control. This study evaluated the effect of Moringa oleifera extract on the cobalt chloride-induced cerebral cortex of adult male wistar rats.  40 male wistar rats weighing (90 ± 120g) were used for the study and they were divided into 5 groups with each group containing 8 rats. Group A served as control which received distilled water, Group B was treated orally with Cobalt chloride at dose 45 mg/kg, Group C received cobalt chloride 45 mg/kg + low dose of Moringa oleifera extract 250 mg/kg for 52 days, Group D treated with cobalt chloride 45 mg/kg + high dose of Moringa oleifera extract 500 mg/kg and Group E treated with 500 mg/kg Moringa oleifera extract only and rats were sacrificed on the 53rd day by cervical dislocation. The brain of each rat was removed and weighed before half was fixed in formol calcium for histological analysis and the second half was used for oxidative stress parameters. The mean body weight of the wistar rats in group C and E increased significantly (P <0.05) while it decreased significantly (P <0.05) in group D. The biochemical analysis shows a significant increase (P<0.05) in the level of MDA in group B and a significant decrease (P<0.05) in group E. additionally, NO level shows a significant increase (P<0.05) in group B compared with control. SDH activity decreased significantly in group C, D, and E. Microscopic examination of the cerebral cortex in group B, C and D showed degenerative changes compared with normal histological features in A and E. The study concluded that cobalt chloride induced cerebral cortical damage while administration of Moringa oleifera extract attenuated the toxic effect of cobalt chloride in wistar rats.


Author(s):  
A. E. Anyabolu ◽  
D. N. Ezejindu ◽  
B. N. Obinwa

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.


Author(s):  
U. S. Aguwa ◽  
C. E. Eze ◽  
B. N. Obinwa ◽  
S. N. Okeke ◽  
S. F. Onwuelingo ◽  
...  

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.


2008 ◽  
Vol 23 (2) ◽  
pp. 179-183 ◽  
Author(s):  
Robson Azevedo Dutra ◽  
Wemberton Martins Araújo ◽  
José Ivan de Andrade

PURPOSE: To assess the effects of Roux-en-Y jejunal limb length on gastric emptying and enterogastric reflux. METHODS: Seventy male Wistar rats were submitted to antrectomy with Roux-en-Y reconstruction and then were divided into two groups of 35 animals. Group A, short limb (7.5 cm) and Group B, standard limb (15 cm). Group A and B were subdivided into five subgroups each in order to study enterogastric reflux at 30 and 60 minutes and to evaluate gastric emptying at 5, 10 and 15 minutes. In order to measure gastric emptying and enterogastric reflux, radiotracers 99m Tc-Phytate and 99m Tc-DISIDA were respectively used. RESULTS: For gastric emptying, the radiotracer concentration was lower in Group A than in Group B after five minutes. The enterogastric reflux was present, but there were no significant differences between enterogastric reflux indexes concerning both A and B Groups. CONCLUSION: A standard Roux limb, besides being unable to protect the stomach from the enterogastric reflux, may become a functional barrier for gastric emptying.


Author(s):  
Carlos Edmundo Rodrigues FONTES ◽  
Ana Paula de ABREU ◽  
Aretuza Zaupa GASPARIM

ABSTRACT Background: Researches on Chagas disease still use several animals and rats, due to size and susceptibility were preferred by many authors. Aim: To develop an experimental model of megacolon in rats inoculated with the strain Y of Trypanosoma cruzi. Methods: Thirty male Wistar rats were distributed in three groups inoculated with different inoculants: Group A: 600000, Group B: 1000000 and Group C: 1500000 blood trypomastigotes of T. cruzi. Animals were sedated intramuscularly at zero inoculation time (T0) and 60 days after inoculation (T60), to perform the barium enema in order to evaluate the dilatation of the different segments of colon in a comparative study of the measurements obtained, using a digital caliper. Evidence of infection was performed by blood smear collected from the animal’s tail 18 days after inoculation with observation of blood forms. Results: Comparing the intestinal diameter of the inoculated animals with 60,0000 trypomastigotes in the T0 of infection with T60 days after the inoculation, significant dilatation was observed between the proximal, medial and distal segments (p<0.01), indicating the establishment of the megacolon model. In addition, comparing intestinal diameter between the different segments, with in the T0 of infection and the T60 after inoculation, significant alterations were observed (p<0.05). Conclusion: The proposed model was possible for in vivo studies of alterations due to infection by T. cruzi and functional alterations of the colon. In addition, the changes manifested in the colon are not directly proportional to the size of the inoculum, but to the time of infection that the animals were submitted, since the animals inoculated with 60,0000 blood forms were the ones which presented the most significant alterations.


2014 ◽  
Vol 31 (01) ◽  
pp. 028-032
Author(s):  
A. Okunlola ◽  
C. Okunlola ◽  
C. Okani ◽  
O. Adewole ◽  
D. Ofusori ◽  
...  

Abstract Introduction: Arteether™, a derivative of artemisinin, is among the recent drugs that have given renewed hope for combating malarial menace. The present study investigated the effects of arteether™ on the histology of the retina and cerebellum of Wistar rats. Materials and Methods: Twenty adult albino Wistar rats weighing 150-200 g, were randomly divided into four groups (A, B, C and D) of five animals each and used for this study. Group A rats were given intramuscular (i.m.) arteether™ (3 mg/kg b.w.) daily for 3 days. Group B rats were given i.m. arteether™ (6 mg/kg b.w.) daily for 3 days. Group C rats were also given i. m. of arteether™ (3 mg/kg b. w.) daily for 3 days, and the same dose was repeated at two-weekly intervals for 4 further weeks; while Group D rats which received normal saline (0.9 % w/v, 3 ml/kg b.w.), served as controls. At the end of the experiment, the rats were sacrificed by cervical dislocation. The retina and cerebellum were excised and processed routinely for histopathology changes, using haematoxylin and eosin stain (H & E), as well as Nissl stain. Results: Results obtained showed normal cellular components of the retina and cerebellum in all groups, and no cyto-pathological changes were observed. Conclusion: Thus, this study showed that under light microscopic examination, therapeutic doses of arteether™ caused no significant cyto-pathologic changes in the retina and cerebellum of Wistar rats.


2018 ◽  
Vol 5 (11) ◽  
pp. 725-737
Author(s):  
Sunday A. Adelakun ◽  
Olusegun D. Omotoso ◽  
Grace T. Akingbade

Globally mercury (Hg) has been reported as one of heavy metal of known toxicity, noted for inducing public health disasters. Present study examines the therapeutics potentials of Ocimum basilicum on mercury chloride (HgCl2) induced hepatotoxicity in Wistar rats. Thirty adult Wistar rats randomly divided into six groups (A-F) of five rats each. Group A served as control was given 2 mL/day of distilled water, Group B, C, D, E and F received 500 mg/kg body weight (bwt) of O. basilicum extract, 20 mg/kg/bwt of HgCl2, 40 mg/kg bwt of HgCl2, 20 mg/kg bwt of HgCl2 and 500 mg/kg bwt O. basilicum leave extract, 40 mg/kg bwt and 500 mg/kg bwt O. basilicum administered daily by gastric gavage, for 21 consecutive days. The gross anatomical parameters of the liver and liver histology were assessed. Liver oxidative stress was evaluated by liver superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH) and malondialdehyde (MDA) assays. The activities of the biomarker enzymes of the liver (alanine transaminase, aspartate transaminase and alkaline phosphatase were assayed). Histological profiles of the liver revealed derangement of the liver cytoarchitecture following consumption of mercury chloride and a marked improvement was observed after O. basilicum administration. Similarly, O. basilicum improved the reduction of antioxidant parameters (SOD, CAT, GPx and GSH) and the increased MDA caused by mercury chloride consumption. O. basilicum thus proffer protection against free radical mediated oxidative stress in mercury chloride-induced hepatotoxicity in rats.


2017 ◽  
Vol 2017 ◽  
pp. 1-7 ◽  
Author(s):  
Ibrahim Semuyaba ◽  
Abass Alao Safiriyu ◽  
Emmanuel Ayikobua Tiyo ◽  
Remón Figueredo Niurka

Studies suggest that garlic (A. sativum) improves memory dependent on the hippocampus. However, the effect of ethanol garlic extract on hippocampus Na+/K+ATPase, Ca2+ATPase, and glutamine synthetase (GS) activities as possible mechanisms in memory improvement in diabetic Wistar rats has not been reported. Twenty-four male Wistar rats weighing 200–250 g were divided into three groups with 8 rats each. Group (A), normal control rats, and Group (B), diabetic rats, received 1 ml of normal saline; diabetic rats in Group (C) received 1000 mg/kg of garlic extract orally for 21 days. Hyperglycemia was induced by a single intraperitoneal injection of streptozotocin 60 mg/kg followed by 120 mg/kg nicotinamide while extraction of garlic was done by cold maceration method. Memory was tested in all groups. After that, the rats were sacrificed, the brain was removed, and the hippocampi were carefully excised and then homogenized. Activities of Na+/K+ATPase, calcium ATPase, and GS were analyzed from the homogenate. Results showed improvement in memory and a significant increase(P<0.05)in hippocampus Na+/K+ATPase, Ca2+ATPase, and GS activities in diabetic rats treated with garlic extract. In conclusion, the increased activity of hippocampus Na+/K+ATPase, calcium ATPase, and glutamine synthetase may account for the memory improvement.


1970 ◽  
Vol 4 (2) ◽  
pp. 44-50
Author(s):  
MB Ekong ◽  
AO Igri ◽  
TB Ekanem

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  


2000 ◽  
Vol 34 (3) ◽  
pp. 252-259 ◽  
Author(s):  
A. B. Criado ◽  
I. A. Gómez de Segura ◽  
F. J. Tendillo ◽  
F. Marsico

Preoperative analgesics are being increasingly used to provide analgesia in the intraoperative and postoperative period. Opioids reduce anaesthetic requirements, although the effect varies with the different drug and species. The aim of this work was to determine whether buprenorphine reduces the minimum alveolar concentration (MAC) of isoflurane in a dose-related fashion, and whether this effect is similar to morphine when clinical doses of both drugs are used in the rat. Thirty-six male Wistar rats were anaesthetized with isoflurane, and MAC was determined before and after the administration of either buprenorphine or morphine. MAC of isoflurane was determined from alveolar gas samples when a standard noxious stimulus, in the form of a tail clamp, was applied. The duration and degree of reduction of the MAC of isoflurane were recorded. Basic cardiovascular and respiratory measurements were also recorded. Buprenorphine (10, 30 and 100 μg/kg) and morphine (1, 3 and 10 mg/kg) reduced in a dose-dependent fashion the MAC of isoflurane by 15%, 30% and 50%, respectively. Buprenorphine resulted in less cardiovascular and respiratory depression and had a longer-lasting action than morphine. In conclusion, buprenorphine has a dose-related isoflurane sparing effect in the rat similar to that caused by morphine at clinical doses of both drugs.


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