Improvement of the Culture Conditions for In Vitro Production of Cattle Embryos in a Portable CO2 Incubator

The effects of different concentrations of growth hormone (GH) on in vitro maturation (IVM), fertilization (IVF) and culture (IVC) of bovine oocyte/embryos in CR1aa or CR2aa media using a simple CO₂ incubator were investigated. The IVM/IVF/IVC of oocytes were carried out in the presence of 0, 50, 100 and 200 ng/ml GH in the medium. The proportion of metaphase II oocytes was significantly higher (p < 0.05) in 200 ng/ml compared with 0 ng/ml GH in CR1aa medium (59 versus 85%, respectively), but this effect was not observed under CR2aa. Higher concentrations of GH yielded lower rates of unfertilized ova and thus superior cleavage rates (36.5 ± 0.2 and 63.5 ± 2.0% versus 17.5 ± 0.2 and 82.5 ± 1.5% or 40.4 ± 0.6 and 59.6 ± 1.4% versus 16.6 ± 1.2 and 83.4 ± 6.2% for 0 and 200 ng/ml GH in portable or ordinary incubator, respectively) in CR1aa. This dose‐dependent effect was also observed in the percentages of transferable embryos, although not statistically different (17.2 ± 1.7 versus 27.3 ± 1.8% and 16.6 ± 3.1 versus 26.0 ± 1.4%, for 0 versus 200 ng/ml GH in portable and ordinary incubator, respectively). In contrast to the CR1aa, different concentrations of GH in CR2aa medium did not increase either fertilization or cleavage rates. In fact, higher concentrations of GH in this medium negatively affected the rate of transferable embryos. Hence, percentages of transferable embryos obtained in the portable incubator under 0 or 50 ng/ml GH were higher (p < 0.05) compared with those obtained in 100 or 200 ng/ml GH (35.4 ± 5.7 or 40.5 ± 5.4% versus 22.4 ± 2.4 or 15.5 ± 2.1%, respectively). There was however, no significant difference in the rate of transferable embryos in an ordinary incubator employing CR2aa medium, but the trend was more or less similar to that observed in the portable incubator. Despite the fact that relatively fewer oocytes were employed for the culture in the ordinary incubator, overall results observed employing the simple portable CO₂ incubator were within the range of those obtained in an ordinary incubator; implying that the simple portable incubator can effectively be employed for the in vitro production of bovine embryos under field conditions.     

An appropriate ingestion volume of oral sulfa drug suspension in pigs

The influence of ingested volume of a sulfa drug suspension, sodium sulfamonomethoxine (SMMNa), on the oral pharmacokinetics was studied in pigs, with regard to bioavailability and gastric emptying. Eighteen pigs, weighing 30-70 kg, were used. Phenol red solution was used for the evaluation of gastric emptying study. SMMNa suspension was used for pharmacokinetic study. Both of these fluids were administered by natural swallowing. Three experimental groups were constructed: G-I; 5 ml/kg of the test fluids to starved animals, G-II; 5 ml/kg of the test fluids to fed animals and G-III; 20 ml/kg of the fluids to fed animals. The glucose glycine electrolyte solution (GGES) was used as the vehicle for both the compounds. Six pigs, having duodenal cannula, were used for the study of gastric emptying. The gastric emptying rate was rapid in G-I, relatively rapid in G-III, and slow and variable in G-II. In agreement with the result of gastric emptying study, the values of Cmax and tmax were high and rapid in G-I, relatively high and rapid in G-III, and low and slow in G-II. Accordingly, the voluminous ingestion of drug suspension can facilitate the gastric emptying, in turn may make the oral absorption of the drug rapid-and-uniform. The 20 ml/kg volume of sulfa drug suspension may practically be recommended for the oral administration in pigs.     

Identification of tropomyosin as a major allergen in the octopus Octopus vulgaris and elucidation of its IgE-binding epitopes

ABSTRACT: The major allergen (named Oct v 1) in the muscle of the octopus Octopus vulgaris was purified by gel filtration on Sephacryl S-300, anion-exchange fast protein liquid chromatography on Mono Q and reverse-phase high-pressure liquid chromatography on TSKgel Octadecyl-4PW. In addition to the molecular mass, amino acid composition and cross-reactivity with Tur c 1 (turban shell Turbo cornutus allergen), the determined partial amino acid sequence clearly demonstrated that Oct v 1 is tropomyosin, similar to the known molluscan and crustacean allergens. Using peptide fragments isolated from the lysylendopeptidase digest of Oct v 1, competitive enzyme-linked immunosorbant assay inhibition experiments showed that IgE-binding epitopes of Oct v 1 are contained in two peptides (77–112 and 148–160) in the central region and one peptide (269–281) in the C-terminal region. In the peptide 77–112, the same sequence as the IgE-binding epitope proposed for Cra g 1 (oyster Crassostrea gigas allergen) is recognized at 92-105. Moreover, the peptide 148-160 partly overlaps with the IgE-binding epitopes suggested for Pen i 1 (shrimp Penaeus indicus allergen) and Pen a 1 (shrimp Penaeus aztecus allergen), and the peptide 269–281 with those for Tur c 1 and Pen a 1.     

Impairment of macrophage function in Mongolian gerbils.

Chemiluminescence studies on superoxide generation by phagosomes using opsonized zymosan showed the highest fluorescence in murine splenic macrophages among four different kinds of splenic or peritoneal macrophages from mice or gerbils. Murine splenic macrophages phagocytized two to three times more latex particles than gerbil splenic macrophages, but peritoneal macrophages did not show a significant difference in phagocytic activity between mice and gerbils. Phagocytosis by macrophages was determined by a technique based on measurement of the release of hydrogen peroxide and myeloperoxidase from phagosomes using microspheres conjugated with 3-(p-hydroxyphenyl) propionic acid (HPPA-MS). HPPA is a substrate of lysosomal myeloperoxidase. The fluorescence of HPPA-HPPA-MS produced by phagocytized HPPA-MS was measured with an immunoreaction analysis system (IMRAS), and the enzyme activities of the four different kinds of peritoneal or splenic macrophages from mice and gerbils were compared. All four kinds of macrophages produced HPPA-HPPA-MS in their phagosomes during phagocytosis and murine splenic macrophages showed the highest level of enzyme activity.     

Protection against chicken leucocytozoonosis provided by immunization with spleen homogenate infected with Leucocytozoon caulleryi

Protection against chicken leucocytozoonosis was assessed in chickens immunized with spleen homogenates from chickens that had received sporozoites of Leucocytozoon caulleryi 7 or 13 days previously. Chickens immunized with the homogenate were challenged with sporozoites of L. caulleryi and observed for changes in clinical signs, parasitemia, serum-soluble antigen, and antibody responses. In chickens immunized with either the 7-day or 13-day homogenate, clinical signs and parasitemia were moderate, mild or absent. This was the case both after immunization with the homogenate and after sporozoite challenge. Immunization with spleen homogenates demonstrated protection against chicken leucocytozoonosis.