Effect of Chemical or Electrical Activation of Bovine Oocytes on Blastocyst Development and Quality

Activation of in vitro‐matured (IVM) oocytes is essential for successful embryo production following nuclear transfer (NT) or intracytoplasmic sperm injection (ICSI). This study was designed to compare the rates of blastocyst production and embryo quality (as measured by numbers of viable cells) following parthenogenetic activation with electrical pulse or the use of two different calcium ionophores, A23187 (CA) or ionomycin (IO), with or without the addition of bovine serum albumin (BSA). IVM oocytes with a first polar body were randomly allocated to five treatment groups: CA (5 μm CA, 5 min; n = 88), CA + BSA (5 μm CA, 5 min; BSA, 5 min; n = 90), IO (5 μm IO, 5 min; n = 91), IO + BSA (5 μm IO, 5 min; BSA, 5 min; n = 86) and EL (two pulses of 1.5 kV/cm, 20 μs; n = 120). Blastocyst rates were higher (p < 0.05) for CA (54.4%), IO (51.4%) and EL (54.5%) than for IO + BSA (18.3%). Treatment CA + BSA (39.8%) did not differ from the others. There was no difference (p > 0.05) among treatments in total number of cells. However, the percentage of viable cells was reduced in CA (49.9%), CA + BSA (45.8%), IO (64.9%), IO + BSA (50.9%) compared with EL (82.7%). In summary, the addition of BSA to the IO treatment had an adverse effect on blastocyst production rates. Although there was no difference between electrical stimulation and chemical activation on blastocyst production rates, electrical activation resulted in blastocysts with a higher percentage of viable cells.     

The detection of CD2+, CD4+, CD8+, and WC1+ T lymphocytes, B cells and macrophages in fixed and paraffin embedded bovine tissue using a range of antigen recovery and signal amplification techniques

In order to develop procedures to label the main bovine leucocyte populations in paraffin embedded sections, the immunoreactivity of 25 monoclonal antibodies (mAbs) to different leucocyte antigens was assessed with formal dichromate (FD5) and 10% formalin fixation, a battery of antigen retrieval (AR) methods, and the biotin-tyramide amplification system. All the leucocyte populations investigated (CD2+, CD4+, CD8+, WC1+ T lymphocytes, B cells and macrophages) were strongly and specifically detectable under an appropriate combination of mAb, AR method and signal amplification system. CD4 and CD8 required the most stringent conditions and could only be demonstrated in FD5 fixed sections. For detection of CD2, WC1+ T lymphocytes, B cells and macrophages, all the mAbs produced immunoreactivity in FD5 or formalin fixed tissues. The need to check a range of different AR methods is stressed, as the method of choice varied for each individual mAb. The incorporation of the signal amplification system was necessary to observe a strong signal and the complete distribution of CD4, CD8 and B cells. Fixation by FD5 proved to be better than formalin for the preservation of surface antigens but it was inferior for the detection of markers which were found to show cytoplasmic immunoreactivity, such as the macrophage marker MAC387 or the B cell markers BAQ155 or IL-A59.     

Genetic and phenotypic aspects of early reproductive performance in Raeini Cashmere goats

This study used pedigree information and data collected from 1979 to 2012 at the Raeini Cashmere goat breeding station, located in Baft City in Kerman Province in southeastern Iran. Genetic and phenotypic parameters for early reproductive traits of breeding does, including total numbers of kids born at first kidding (LSB1), total numbers of kids weaned at first kidding (LSW1), total birth weight of all kids born at first kidding (LWB1), total weaning weight of all kids weaned at first kidding (LWW1), and age at first kidding (AFK), were estimated using a Bayesian approach via Gibbs sampling. Posterior means for heritability estimates of LSB1, LSW1, LWB1, LWW1, and AFK were statistically significant, with values of 0.12, 0.23, 0.17, 0.15, and 0.46, respectively. Low-to-moderate additive genetic variation was present for the studied reproductive traits. Estimated genetic correlations among LSB1, LSW1, LWB1, and LWW1 were statistically significant and ranged from 0.12 between LWB1 and LWW1 to 0.72 between LSB1 and LSW1. Corresponding phenotypic correlation estimates were also statistically significant and ranged from 0.04 between LWB1 and LWW1 to 0.55 between LSB1 and LSW1. Posterior means of genetic and phenotypic correlations between AFK and other studied traits were statistically significant only for LSB1 and LWB1. For LSB1, LSW1, LWB1, and LWW1, we conclude that genetic and phenotypic improvement in any of these traits in Raeini Cashmere does would favorably influence all of the other traits. However, does that first kidded at younger ages have smaller litters at birth and lower litter birth weights at their first parity.

     

The histology of dermal glands of mating Breviceps with comments on their possible functional value in microhylids (Amphibia: Anura)

During mating male and female Breviceps become ‘glued’ together. The distribution of multicellular dermal glands varies between the sexes. The mate has a large number of holocrine glands on the ventrum, while the female has similar glands on the dorsum of the back. II thus appears that both sexes contribute to the ‘glueing’ mechanism. New hypotheses to explain the adhesion between the sexes are proposed, viz., that the shift to terrestrial habils and subterranean laying in Breviceps have secondarily resulted in male si2e diminution. In other microhylids which adhere during aquatic oviposition, yet demonstrate a terrestrial morphology and habits, bouyancy is suggested to be of primary functional value.     

Cross-protective immunity and the serological classification system for Bacteroides nodosus

The relationship between the serological classification system for serogroup B and for serogroup H of Bacteroides nodosus and cross-protection between subgroups within these serogroups was examined. Protection against ovine footrot following vaccination was achieved against other subgroup strains provided sufficient cross-reactive antibody was induced by shared pilus antigens. Within serogroup B, better cross-protection against one subgroup was obtained with a pili vaccine than a whole cell vaccine which correlated with higher pilus antibody titres induced by the former. For serogroup H, a lack of cross-protection and serological reactivity between subgroups was demonstrated, which indicates that the prototype strain of subgroup H2 should be designated a new serogroup.     

Relationship of blood corticosterone, immunoglobulin and haematological values in young crocodiles (Crocodylus porosus) to water temperature, clutch of origin and body weight

Objective To examine whether sub-optimal temperature induced stress and immunosuppression in farmed saltwater crocodile (Crocodylus porosus) hatchlings. Design A clinico-pathological study. Animals A total of 140 hatchlings were used. Procedure Body weight and length, plasma corticosterone and immunoglobulin concentrations and total and differential white blood cell counts were measured in 140 hatchlings from five clutches divided between five water temperature treatment groups. Initially all groups were housed at 32°C for 10 weeks, then two groups (L, LC) were changed to low temperature (28°C) and two groups (H, HC) to high temperature (36°C), while one group (C) remained at 32°C. The LC and HC groups were maintained at these temperatures for 10 days, after which the water temperature of both groups was returned to 32°C. Blood samples were collected twice (at 6 and 9 weeks of age) before the initial temperature change, and at 10 days and 4 weeks after the initial temperature change (at 11.5 and 14 weeks of age). Results Except for an increase in plasma corticosterone in the HC group and a decrease in the L group when the temperature change was first introduced, changes in plasma corticosterone were not significant. There were no significant changes in immunoglobulin concentrations. There were, however, significant decreases in the total white cell and lymphocyte counts in the LC group after the temperature was decreased to 28°C, and an increase in these counts after water temperature was returned to 32°C. Clutch of origin had significant effects on body weight and length gains, and there were negative relationships between body weight and corticosterone concentrations and between body weight and immunoglobulin concentrations. Conclusions As haematological changes indicative of stress were not associated with significant changes in serum corticosterone, immunosuppression in young crocodiles may be independent of the hypothalamic-pituitary-adrenal cortical axis.