Effects of Hormonal Supplementation on Nuclear Maturation and Cortical Granules Distribution of Canine Oocytes During Various Reproductive Stages

The aim of this study was to evaluate the effects of hCG, progesterone and oestradiol supplementation on nuclear and cytoplasmic maturation of canine oocytes cultured for 24, 48, 72 and 96 h. Oocytes obtained from 18 healthy bitches were divided into three groups according to their reproductive status (follicular, luteal and anoestrus stages) and cultured in TCM 199 + 25 UI/ml of hCG + 1 μg/ml of progesterone + 1 μg/ml of 17‐β oestradiol or without hormonal supplementation (control) for different periods. Then, they were stained with FITC‐LCA‐Hoescht for chromatin configuration and cortical granules distribution and evaluated under an epifluorescence microscope. Culture time and the influence of different stages of the oestrous cycle were also evaluated. The present study demonstrated that there was no significant difference among the reproductive stages. With regards to culture medium, only oocytes from the supplemented medium were able to complete meiosis; however, significant difference was only noticed in the percentage of MI stage oocytes (p < 0.05) in the follicular and luteal group at 72 h of culture. Most oocytes in germinal vesicle, germinal vesicle breakdown and metaphase I stage had cortical granules distributed throughout the cytoplasm (immature pattern), irrespective of the culture period (p < 0.05). Cortical granules distributed immediately beneath the plasma membrane (mature) was only observed in metaphase II stage oocytes, but not all of them presented matured cytoplasm. Our results reveal that cortical granules distribution in canine oocytes matured in vitro did not progressed in correspondence with nuclear stage changes and are in accordance with those from other species.     

Immunoglobulin M and secretory immunoglobulin A: presence of a common polypeptide chain different from light chains

Unique polypeptide chains have been isolated from S-sulfonated light-chain fractions of human serum immunoglobulin M and colostral immunoglobulin A. Their electrophoretic mobilities, molecular weights, peptide maps, amino acid compositions, and antigenic determinants are very similar or perhaps identical but differ from those of light chains and secretory piece.     

Dose titration of an injectable formulation of lufenuron in cats experimentally infested with fleas

OBJECTIVES: To identify the lowest single dose of lufenuron injected s.c. that results in a 90% disruption of the flea (Ctenocephalides felis) life cycle for 6 months in cats. ANIMALS: 40 domestic shorthair cats (20 males, 20 females) between 5 and 7 months old. PROCEDURE: Cats were randomly assigned to 1 of 5 eight-cat groups and experimentally infested with C. felis on days -8, -7, -6, and -4. On day 0, cats in the 4 treatment groups were treated with an injectable formulation of lufenuron at doses of 2.5, 5, 10, or 20 mg/kg of body weight, respectively. Control cats received the injectable formulation without lufenuron. Experimental infestations were repeated and flea eggs collected at various intervals for 196 days after treatment. Eggs were placed in media and incubated in an insectary for 28 days to determine effects of injectable lufenuron on egg and larval development. Number of adults that emerged from eggs were compared among groups. RESULTS: Lufenuron injected once at a dose of 10 or 20 mg/kg, but not at 2.5 or 5 mg/kg, resulted in a 90% decrease in number of adult fleas emerging from eggs for 196 days after treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that control of flea egg and larval development for at least 6 months can be achieved in cats with a single s.c. injection of lufenuron (10 mg/kg). The injectable formulation may provide veterinarians and cat owners an alternative to the tablet formulation of lufenuron.     

The effects of bovine growth hormone and thyroxine on growth rate and carcass measurements in lambs

The effects of bovine growth hormone (GH) and thyroxine (T4) on growth and carcass characteristics were assessed in Dorset ram lambs. Lambs in four groups (n = 10/group) were treated for 30 d as follows: controls, 3.33 mg (6 IU) GH/d (s.c.); 5-mg T4 implant (s.c.) on d 1 and a 10-mg T4 implant 21 d later; GH + T4. Blood samples were collected at 3-d intervals for analysis of GH, T4, triiodothyronine, somatomedin-C and testosterone concentrations. Six lambs/group were slaughtered for carcass measurements and composition. Daily GH injections increased (P less than .005) baseline plasma GH levels 10-fold, whereas plasma T4 concentrations were increased 10% (P less than .10) by the implants. Somatomedin-C increased with time in all groups, but the increments from d 0 to d 30 were higher (P less than .05) with GH treatment. Average daily gain (mean = 352 g/d), feed consumption and feed to gain ratio were not affected (P greater than .1) by GH or T4 treatment in ram lambs. Hot carcass weight and dressing percentage were increased (P less than .05) by T4. Growth hormone increased carcass protein content (P less than .005) and muscle weights while reducing carcass fat (P less than .05). Carcass composition was not altered by T4 alone, and the T4 x GH interaction was not significant; however, the combination of T4 and GH resulted in greater muscle and protein weight than did either hormone alone or no hormone administration. There were no differences in bone length or in the metacarpal growth plate width among groups. The beneficial effects of GH on carcass composition were not further enhanced by administration of thyroxine.     

Production of ovine chimeras by inner cell mass transplantation

Ovine chimeras were produced by micro-injection of isolated inner cell masses (ICM) into recipient blastocysts. Inner cell masses were isolated by immunosurgery. A total of 57 chimeric embryos was produced, 52 of which were transferred to recipient ewes. Thirty-seven live lambs were born, of which 15 were determined to be chimeric on the basis of blood type analysis. One lamb, although not a blood chimera, exhibited overt signs of chimerism. An additional six lambs were determined to have developed solely from the injected ICM. The rate of chimerism in live lambs was 43% (16/37) while the survival rate of injected ICM was 59% (22/37). The method presented allows the production of relatively large proportion of viable, chimeric embryos without the use of an intermediate recipient.     

Magnesium Oxide Induced Metabolic Alkalosis in Cattle

A study was designed to compare the metabolic alkalosis produced in cattle from the use of an antacid (magnesium oxide) and a saline cathartic (magnesium sulphate). Six, mature, normal cattle were treated orally with a magnesium oxide (MgO) product and one week later given a comparable cathartic dose of magnesium sulphate (MgSO₄).

The mean percent dry matter content of the cattle feces changed significantly (P<0.001) following administration of both MgO (15.6-8.1) and MgSO₄ (17.0-8.7) but there was no significant difference between treatments. The mean rumen pH values changed significantly (P<0.001) following administration of both MgO (7.-8.7) and MgSO₄ (7.3-8.3) but there was no significant difference between treatments. However, use of the MgO product caused a more severe (P<0.001) metabolic alkalosis as determined by base excess values. The base excess values remained elevated for 24 hours in the MgO treated group compared to only 12 hours after MgSO₄ administration. Following MgO administration, mean hydrogen ion concentration (pH), bicarbonate ion concentration ([HCO₃-]) and base excess were 7.44, 33.3 mmol/L and +8.0 respectively compared to 7.38, 27 mmol/L and +3.0 after MgSO₄.

Since the oral use of MgO in normal cattle causes a greater and more prolonged metabolic alkalosis compared to MgSO₄, MgO is contraindicated as a cathartic in normal cattle or in cattle with abomasal abnormalities characterized by pyloric obstruction and metabolic alkalosis.

     

Production of sheep-goat chimeras by inner cell mass transplantation

Embryos were surgically flushed from goats and sheep on d 6 and 7, respectively, following the first day of estrus (d 0). After enzymatic removal of the zonae pellucidae, inner cell masses were isolated from caprine blastocysts by immunosurgery. The intact inner cell masses were injected into ovine blastocysts with the aid of a micromanipulator. Twenty-two manipulated blastocysts were surgically transferred into 12 ovine recipients. Nine ewes gave birth to a total of 13 young (59% embryo survival). Ten were classified by serum electrophoretic assays or karyotypes as lambs, one as a kid, and two as interspecific chimeras.