A.B.Z. Zuki

Effect of freeze-drying and gamma irradiation on biomechanical properties of bovine pericardium

Freeze-drying and gamma irradiation are the techniques widely use in tissue banking for preservation and sterilization of tissue grafts respectively. However, the effect of these techniques on biomechanical properties of bovine pericardium is poorly known. A total of 300 strips of bovine pericardium each measured 4 cm × 1 cm were used in this study to evaluate the effect of freeze-drying on biomechanical properties of fresh bovine pericardium and the effect of gamma irradiation on biomechanical properties of freeze-dried bovine pericardium. The strips were divided into three equal groups, which consist of 100 strips each group. The three groups were fresh bovine pericardium, freeze-dried bovine pericardium and irradiated freeze-dried bovine pericardium. The biomechanical properties of the pericardial strips were measured by a computer controlled instron tensiometer while the strips thickness was measured by Mitutoyo thickness gauge. The results of the study revealed that freeze-drying has no significant (p > 0.05) effect on the tensile strength, Young’s modulus (stiffness) and elongation rate of fresh bovine pericardium. Irradiation with 25 kGy gamma rays caused significant decreased in the tensile strength, Young’s modulus and elongation rate of the freeze-dried pericardium. However, gamma irradiation has no significant effect on the thickness of freeze-dried bovine pericardium, while freeze-drying caused significant decreased in the thickness of the fresh bovine pericardium. The outcome of this study demonstrated that freeze-drying has no significant effect on the biomechanical properties of fresh bovine pericardium, and gamma irradiation caused significant effect on the biomechanical properties of freeze-dried bovine pericardium.

Distribution of lectin-bindings in the testis of the lesser mouse deer, Tragulus javanicus.

The distribution of lectin bindings in the testis of the smallest ruminant, lesser mouse deer (Tragulus javanicus), was studied using 12 biotinylated lectins specific for d‐galactose (peanut agglutinin PNA, Ricinus communis agglutinin RCA I), N‐acetyl‐d‐galactosamine (Dolichos biflorus agglutinin DBA, Vicia villosa agglutinin VVA, Soybean agglutinin SBA), N‐acetyl‐d‐glucosamine and sialic acid (wheat germ agglutinin WGA, s‐WGA), d‐mannose and d‐glucose (Lens culinaris agglutinin LCA, Pisum sativum agglutinin PSA, Concanavalin A Con A), l‐fucose (Ulex europaeus agglutinin UEA I), and oligosaccharide (Phaseolus vulgaris agglutinin PHA‐E) sugar residues. In Golgi‐, cap‐, and acrosome‐phase spermatids, lectin‐bindings were found in the acrosome (PNA, RCA I, VVA, SBA, WGA and s‐WGA), and in the cytoplasm (PNA, RCA I, VVA, SBA, WGA, LCA, PSA, Con A and PHA‐E). s‐WGA binding was confined to the spermatid acrosome, but other lectins were also observed in spermatocytes. In spermatogonia, VVA, WGA, Con A, and PHA‐E bindings were observed. Sertoli cells were intensely stained with DBA and Con A, and weakly with PHA‐E. In interstitial Leydig cells, RCA I, DBA, VVA, Con A, PSA, LCA, WGA and PHA‐E were positive. UEA I was negative in all cell types including spermatogenic cells. Unusual distribution of lectin‐bindings noted in the testis of lesser mouse deer included the limited distribution of s‐WGA only in the spermatid acrosome, the distribution of DBA in Sertoli cells, Leydig cells and lamina propria, and the absence of UEA I in all type cells. The present results were discussed in comparison with those of other animals and their possible functional implications.

Immunohistochemical Study of Endocrine Cells in the Gastrointestinal Tract of the Barking Deer, Muntiacus muntjak

With 6 figures and 2 tables

Morphological study of pancreatic duct in red jungle fowl

Although the delayed-type hypersensitivity (DTH) skin reaction to tuberculin is used worldwide for tuberculosis (TB) detection, it has poor diagnostic specificity due to the presence of common antigens in tuberculin shared by many mycobacterial species. The problem is noticed, especially in countries where the Bacillus Calmette-Gue´rin (BCG) vaccination is widely practiced. Thus, a new skin test antigen specific for the diagnosis of Mycobacterium tuberculosis (MTB) infection is urgently needed. CFP-10, a mycobacterial secretary protein that is absent in Mycobacterium bovis BCG and most other mycobacterial species including Mycobacterium avium, Mycobacterium intracellulare, has been shown to elicit cellular immune responses in MTB infected individuals and can be a good candidate for MTB specific diagnosis. We prepared recombinant MTB CFP-10, rCFP-10, and its utility as specific antigen for TB diagnosis was evaluated by skin testing in guinea pigs sensitized with M . tuberculosis, M. bovis, and M. bovis BCG. Our results show that the purified MTB rCFP-10 antigen elicits a positive skin response only in the guinea pigs sensitized with M. tuberculosis and M. bovis , and not in the animals sensitized with M. bovis BCG vaccine. The data presented in this study supports further testing of the use rCFP-10 as the specific antigen in the skin test for the diagnosis of MTB infection in humans. Key words : Recombinant CFP-10 protein, skin test, delayed-type hypersensitivity, tuberculosis infection, Mycobacterium tuberculosis, Mycobacterium bovis, Bacillus Calmette-Gue´rin.

Effect of preservation methods on the performance of bovine pericardium graft in a rat model.

This study investigates the effect of preservation methods on the performance of bovine parietal pericardium grafts in a rat model. Mid‐ventral full thickness abdominal wall defects of 3 × 2.5 cm in size were created in 90 male Sprague–Dawley rats (300–400 g), which were divided into three groups of 30 rats each. The abdominal defects of group one and two were repaired with lyophilized and glycerolized bovine pericardium grafts, while the defects of group three were repaired with expanded polytetrafluoroethylene (ePTFE) Mycro Mesh as a positive control. Another group of 30 rats underwent sham operation and was used for comparison as negative control. Each group of rats (n = 30) was divided into five subgroups (n = 6) and killed at 1, 3, 6, 9 and 18 weeks post‐surgery for gross and morphological evaluations. The rats tolerated the surgical procedure well with a total mortality of 0.05%. No serious post‐operative clinical complications or signs of rejection were encountered. Adhesions between the grafts and the underlying visceral organs observed in the study were mostly results of post‐surgical complications. Glycerol preservation delayed degradation and replacement of the grafts, whereas lyophilization caused early resorption and replacement of the grafts. The glycerolized grafts were replaced with thick dense fibrous tissue, and the lyophilized grafts were replaced with thin loose fibrous tissue. The healing characteristic of the bovine pericardium grafts was similar to those of the sham‐operated group, and quite different from those of the ePTFE Mycro Mesh. The outcome of the present study confirmed the superiority of glycerolized bovine pericardium grafts over its lyophilized counter part.

Effects of feeding whole linseed on ruminal fatty acid composition and microbial population in goats

The objective of the present study was to evaluate the effect of feeding different levels of whole linseed, as a source of n-3 polyunsaturated fatty acids (PUFA), on ruminal fatty acid composition and microbial population in the goat. Twenty-four crossbred Boer goats were assigned to 3 dietary treatments: L0 (control), L10 and L20 containing 0, 10%, or 20% whole linseed, respectively. The ruminal pH and concentration of total volatile fatty acids (VFA) were not affected by dietary treatments. The feeding of L10 and L20 diets produced higher (P < 0.05) molar proportions of acetate and lower (P < 0.05) molar proportions of butyrate and valerate than the L0 diet. Molar proportions of myristic acid (C14:0) and palmitic acid (C16:0) were lower (P < 0.05) in the rumen of goats offered L10 and L20 diets than the control diet. However, stearic acid (C18:0), vaccenic acid (C18:1 trans-11), conjugated linoleic acid (CLA, C18:2 trans-10, cis-12) and α-lenolenic acid (C18:3 n-3) were higher (P < 0.05) in the rumen of goats fed L10 and L20 than L0. Both inclusion levels of linseed in the diet (L10 and L20) reduced the ruminal total bacteria, methanogens, and protozoa compared with L0 (P < 0.05). The effect of the dietary treatments on cellulolytic bacteria, varied between the individual species. Both inclusion levels of linseed resulted in a significant decrease (P < 0.05) in the population of Fibrobacter succinogenes, and Rumunococus flavefaciens compared with L0, with no significant difference between the groups fed linseed diets. The population of Rumunococus albus was not affected by the different dietary treatments. It was concluded that inclusion of whole linseed in the diet of goats could increase the concentration of PUFA in the rumen, and decrease the population of F. succinogenes, R. flavefaciens, methanogens and protozoa in rumen liquid of goats.

Activities of amylase, trypsin and chymotrypsin of pancreas and small intestinal contents in the red jungle fowl and broiler breed

In order to reduce the cost and time of in vitro raised plants of Trichosanthes dioica Roxb., a minimal medium has been formulated by substituting costly growth regulators from the medium with a cost effective constituent, the coconut milk. A semisolid Murashige and Skoogs’s medium supplemented exclusively with 15% coconut milk showed the highest percentage of plantlet regeneration (99%) in the explants. When nodal, shoot-tip and immature leaf explants were cultured on this medium, rhizogenesis was observed in about 5 to 6 days of inoculation, followed by shoot formation in about 8 to 10 days. The fully developed plantlets, 10 to 12 cm in length with professed roots were obtained in about 20 days of inoculation in a single step without adding/changing growth regulators. After transplantation in the potted soil, these plantlets showed similar growth patterns as compared to the plants obtained from a conventional three-four step method of tissue culture experiments. Keywords: Coconut milk, tissue culture, Trichosanthes dioica Roxb

Histomorphometric evaluation of small intestinal mucosa of red jungle fowl and commercial broiler from one day to four months of age

Histomorphometry of the small intestinal mucosa of the red jungle fowl (RJF) and commercial broiler breed (CBC) from day one to four months post-hatch were investigated. For the sake of comparison between these two breeds, the following parameters were included: the number of villi, villus surface area and the intestinal surface area for each small intestinal segment. New procedure for enumerating the intestinal villi was performed by using the standard paraffin sections, whereby the villi were counted through their cross sections using an image analyzer. The numbers of villi were significantly higher in the RJF than the CBC for all the intestinal segments from the day one to four months posthatch. The villi numbers showed a decreased with age. The villus surface area was significantly higher in the CBC than the RJF for all intestinal segments for all the ages. For the duodenal surface area, the RJF showed a higher value than the CBC at one day old chick, but data was reversed on day 20 posthatch; the data in the remaining days showed no significant differences. While for the jejunal and ileal surface areas, the CBC exhibited significantly higher values except day 10 where both breeds showed no difference for the jejunum and day 120 for the ileum, the RJF showed higher value.The selection for high body weight decreased the villi number of the small intestine, but in the same time increased the villus surface area for all the intestinal segments which reflects the increase in mucosal surface area particularly the jejunum and ileum. Key words : Intestinal villi, villi number, surface area, red jungle fowl.

Secretary IgA concentrations and plasma cell count changes associated with the estrous cycle in ewes

Problems statement: The level of uterine Secretory-IgA (S-IgA) and num bers of plasma cells was measured to observe the differences betwe en two stages of estrous cycle (follicular and luteal phase) in the healthy cycling non pregnant e wes. Approach: Twelve ewes were used in this study and they were divided into two groups of 6 an imals each according to the stages of estrous cycle. All ewes were subjected to estrous synchroni zation and allowed to undergo one natural estrous cycle after the removal of the sponge. All animals were then slaughtered at the end of the experiment. The uterine mucus was collected by flushing with a mixture of protease inhibitor cocktail in distilled water. For both stages, the level of uterine S-IgA was quantified by using ELISA and Methyl Green Pyronine staining was used to observe the plasma ce ll in the tissues of the uterine horn and oviduct o f ewes genital tract. Results: The results were analyzed by independent sample t-t est and presented as mean ±SEM. This study showed the relationship of the estr ous cycle stages to uterine S-IgA concentration (µg mL -1 ) and populations of plasma cell in the healthy non -pregnant cycling ewes. The concentration (µg mL -1 ) of S-IgA (0.20±0.01) in the follicular phase was highly significant (p<0.01) as compared with the luteal phase (0.17±0.002). In add ition, the populations of the plasma cells were significantly higher (p<0.01) in the uterine horn ( 4.97 ±0.32) and oviduct (3.82 ±0.33) during follicular phase compared to the luteal phase (3.87 ±0.30) and (1.90 ±0.21), respectively. Conclusion: The main reason for the immunosuppression during the luteal phase did not fully justified, especially with the presence of potential acquired infection during coi tus in the follicular phase and at the same time immune system should decrease accordingly to prevent newly attached fetus rejection by the mother immune system.