2008 - Volume 35 - Issue 1

5-8
Technical Report: Mouse Fetal Blood Collection. Taking the Best out of the Old Needle-Syringe Method
by Maria de Lurdes Pinto, Paula Rodrigues, Ana Cláudia Coelho, Luís Antunes, Carlos Gonçalves & Vasco Bairos

Collection of mice blood is not a common practice due to technical difficulties and the small volumes collected. This constrains many researchers that often choose indirect types of sampling to follow fetal and gestational development, diagnosis and therapy. This study is part of wider studies, in which the above technique was optimized in order to obtain larger volumes. Timed-pregnant ICR (CD-1®) mice were used. After anesthesia fetuses were rapidly delivered by hysterectomy at gestational days 15, 16, 17 and 18. Large vessels in the fetuses thorax were cut, and blood was collected using several different techniques: a) capillary tubes, b) Microvette® CB 300 (Sarstedt), c) micropipetes, d) different gauge needles and syringe, and e) a hand-modified 20G needle applied to a 1ml/100 IU syringe. The volumes obtained with the modified 20G needle at 15, 16, 17 and 18 gestational days ranged respectively from: 30 to 45, 50 to 70, 80 to 100, and 120 to 140 μl. The optimization of this technique allows the measurement of biochemical parameters during fetal development and may help to reduce the number of animals used for similar procedures.

13-24
Helicobacter ganmani Infection Associated with a Spontaneous Outbreak of Inflammatory Bowel-like Disease in an IL-10-Deficient Mouse Colony
by Ingrid Nilsson, Erik Sturegård, Björn Barup, Roger Willén, Waleed Abu Al-Soud, Anna Hultberg, Lennart Hammarström, Hans-Olof Nilsson & Torkel Wadström

Background: A breeding colony of IL-10 deficient B6.129P2-Il10 tm1Cgn/J mice, kept under conventional conditions, developed an inflammatory bowel-like disease (IBD) with rectal prolapse and blood tinged diarrhoea. No clinical signs of disease were observed at the time of arrival to our animal house. These animals were originally planned to serve as a negative control group in an experimental infection study with Helicobacter species to investigate colonization of the murine gut. Results: A spiral-shaped, Gram-negative bacterium was isolated from the breeding mice colony. In a first group of six animals, tissue specimens from the liver, small and large intestines, faeces and blood, were analysed by culture, PCR-denaturing gradient gel electrophoresis (PCR-DGGE), species-specific PCR assays and DNA-sequencing, histology and serology. Helicobacter ganmani, but no other Helicobacter species, was isolated from the liver, small bowel, caecum, colon and faeces. We found inflammation in caeca, colon and livers, most pronounced in the caecal areas of culture positive mice with a severe typhlitis with cystic dilatation of glandular structures and irregular crypt architecture. Some animals showed a pronounced colitis with mucosal and sub-mucosal inflammatory infiltrates. Other animals displayed large lymphoid infiltrates in the livers and hepatitis. Tissue samples and sera from 18 additional animals from the same breeding colony were analysed by the same methods, except for culture. H. ganmani was identified by PCR in most tissue samples of the 18 additional animals as well. Sero-conversion to H. ganmani correlated well with histopathological changes. Conclusions: Our findings emphasize the importance of using Helicobacter-free animals to develop murine models of chronic hepatitis and colitis.

29-34
Antidiabetic and Hypolipidemic Effects of Aqueous Methanolic Extract of Acacia Nilotica Pods in Alloxan-Induced Diabetic Rabbits
by Maqsood Ahmad, Fatima Zaman, Tanveer Sharif & Muhammad Zabta Ch.

This study was designed to investigate the effect of an aqueous methanol extract of Acacia nilotica pods (Anp) on various biochemical parameters, namely blood glucose levels, total cholesterol, High density lipids (HDLs), triglycerides, Serum Glutamate Oxaloacetate and Pyruvate Transaminase (SGOT, SGPT) and serum creatinine clearance in alloxan-induced diabetic rabbits. Rabbits were divided into three experimental groups: control, diabetic and Anp treated. The Anp treated group was further subdivided into three different groups based on the dose administered. This showed that a dose of 400 mg/kg body weight maximally reduced the blood glucose levels as compared to the diabetic group (p<0.001). This dose also significantly (p<0.05) lowered the plasma total cholesterol, triglyceride and Low-density lipids (LDLs) in treated rabbits as compared to diabetic rabbits. Furthermore, the same dose also significantly increased the plasma HDL levels of the treated group when compared with the diabetic group. Whereas the activity of SGOT and SGPT were decreased significantly (p<0.001). Anp extract in treated diabetic rabbits. Anp treatment showed no significant effect on creatinine clearance. For interest a paper with similar aims, but using water extract of Nigella stiva L. appeared in this journal in 2004, (Merel et al, 31 (1), 49-53).

39-43
Corneal Thickness in Pigs Measured by Ultrasound Pachymetry In Vivo
by Carsten Faber, Erik Scherfig, Jan Ulrik Prause & Knud Erik Sørensen

The objective was to evaluate the normal thickness and diameter of the pig cornea using, under general anesthesia, 13 female, young, domestic Danish Landrace pigs. The thickness was measured, for the first time in vivo, and were: 666 μm centrally, 657 μm nasally, 713 μm inferiorly, 669 μm temporally, and 714μm superiorly (mean values). The cornea diameters were also measured, using calipers, and were: 14.9 mm horizontally and 12.4 mm vertically (mean values). Conclusion: The pig cornea is only slightly thicker than the human cornea. However, unlike the human cornea, no significant difference in thickness exists between the central and the tempero-nasal peripheral locations of the pig cornea. The pig could prove useful as source for corneas in human xenotransplantation.

45-49
Electrocardiographic Changes Induced by Ivermectin in Guinea Pigs
by M. Sireli, V. Sagmanligil, N Çetin & B. Emre

Guinea pigs were used to observe the possible side effects of ivermectin (500 μg/kg b.w.) on the heart by recording their electrocardiograms (ECG) at 24, 48 and 72 hours after its injection (experimental groups). Significant effects of ivermectin on the electrocardiographic parameters belong to limb lead II were compared to those of the control group. These were: the increases in amplitudes of P, S (only at 24 hours) and T waves, and also the lengthening (or “prolongation”) in the durations of P and T waves, QRS complex (only at 24 and 48 hours) and PR and QTc intervals at 24, 48 and 72 hours. As a consequence of the prolonged durations mentioned above, statistically significant decreases of heart beats were found on the electrocardiograms recorded at 24 and 48 hours after ivermectin injection (p<0.05). In conclusion, the weak conductance in the heart of guinea pigs as an indicator of the prolongation in durations and intervals (negatively dromotropic) induced by ivermectin can be interpreted as its inhibitory effect on the sarcoplasmic reticulum Ca2+ -ATPase pump causing a late Ca2+ accumulation after systole due to the reduced amount of Ca2+.

53-56
Correlation Between The Erythrocyte Sedimentation Rate and Blood Nitric Oxide Levels in Rabbits?
by Metehan Uzun, Sinan Saral, Onur Atakisi, Kursad Yapar, Erdogan Uzlu, Mehmet Citil,
Didem Tastekin & Hidayet Metin Erdogan

This study was designed to determine whether elevated or decreased blood nitric oxide (NOx) values following injections of L-arginine and N-nitroarginine methylester (L-NAME) are positively correlated with the erythrocyte sedimentation rate (ESR) in healthy rabbits. The study involved 21 New Zealand rabbits of both sexes, aged between 10 and 15 months old. They were divided into 3 equal groups. The control group received 1 ml isotonic saline per rabbit, and Groups I and II received 100 mg/kg L-NAME and 250 mg/kg L-arginine intraperitoneally, respectively. After two hours blood samples were obtained and ESR (30, 60, and 120 minutes) and NOx were estimated. There were no significant differences between the ESR values at all times; however significant correlation between the NOx and ESR existed in Group II at 2 hours after injection. In conclusion, there was a positive correlation between high blood NOx and ESR values, but detailed studies are needed to disclose this correlation.