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.     

Leptin in horses: tissue localization and relationship between peripheral concentrations of leptin and body condition

Obesity has been a major concern in the horse industry for many years, and the recent discovery of leptin and leptin receptors in numerous nonequine species has provided a basis for new approaches to study this problem in equine. The objectives were to: 1) clone a partial sequence ofthe equine leptin and leptin receptor genes so as to enable the design of primers for RT-PCR determination of leptin and leptin receptor gene presence and distribution in tissues, 2) develop a radioimmunoassay to quantify peripheral concentrations of leptin in equine, 3) determine if peripheral concentrations of leptin correlate with body condition scores in equine, and 4) determine if changing body condition scores would influence peripheral concentrations of leptin in equine. In Experiment 1, equine leptin (GenBank accession number AF179275) and the long-form of the equine leptin receptor (GenBank accession number AF139663) genes were partially sequenced. Equine leptin receptor mRNA was detected in liver, lung, testis, ovary, choroid plexus, hypothalamus, and subcutaneous adipose tissues using RT-PCR. In Experiment 2, 71 horses were categorized by gender, age, and body condition score and blood samples were collected. Sera were assayed for leptin using a heterologous leptin radioimmunoassay developed for equine sera. Serum concentrations of leptin increased in horses with body condition score (1 = thin to 9 = fat; r = 0.64; P = 0.0001). Furthermore, serum concentrations of leptin were greater in geldings and stallions than in mares (P = 0.0002), and tended to increase with age of the animal (P = 0.08). In Experiment 3, blood samples, body weights, and body condition scores were collected every 14 d from 18 pony mares assigned to gain or lose weight over a 14-wk interval based on initial body condition score. Although statistical changes (P = 0.001) in body condition scores were achieved, congruent statistical changes in peripheral concentrations of leptin were not observed, likely due to the small range of change that occurred. Nonetheless, serum concentrations of leptin tended to be greater in fat-restricted mares than in thin-supplemented mares (P = 0.09). We conclude that leptin and leptin receptors are present in equine tissues and that peripheral concentrations of leptin reflect a significant influence of fat mass in equine.     

Application of broken-line analysis to assess floor space requirements of nursery and grower-finisher pigs expressed on an allometric basis

Few issues in swine production are as complex as floor space allowances. One method for pork producers to calculate floor space allowance (A) is to convert BW into a 2-dimensional concept yielding an expression of A = k * BW(0.667). Data on ADG, ADFI, and G:F were obtained from published peer-reviewed studies. Five data sets were created: A = grower-finisher pigs, fully slatted floors, and consistent group size; B = grower-finisher pigs and fully slatted floors (group size did not need to be consistent); C = grower-finisher pigs, partially slatted floors, and consistent group size; D = grower-finisher pigs, partially slatted floors (group size did not need to be consistent); and E = nursery pigs, fully slatted or woven wire floors (group size did not need to be consistent). Each data set was analyzed using a broken-line analysis and a linear regression. For the broken-line analyses, the critical k value, below which a decrease in ADG occurred, varied from 0.0317 to 0.0348. In all cases the effect of space allowance on ADG was significant (P < 0.05). Using the linear analyses based on data with k values of < 0.030, the critical k values for the 4 grower-finisher data sets did not differ from those obtained using the broken-line analysis (0.0358 vs. 0.0336, respectively; P > 0.10); however, none of the linear regressions explained a significant proportion of the variation in ADG. The slopes for the nonplateau portion of the broken-line analyses based on percent values varied among data sets. For every 0.001 decrease in k (approximately 3% of the critical k value), ADG decreased by 0.56 to 1.41%, with an average value of 0.98% for the 5%-based analyses. The use of an allometric approach to express space allowance and broken-line analysis to establish space requirements seem to be useful tools for pig production. The critical k value at which crowding becomes detrimental to the growth of the pig is similar in full- and partial-slat systems and in nursery and grower-finisher stages. The critical point for crowding determined in these analyses approximated current recommendations to ensure the welfare of pigs.     

Field Test of Digital Photography Biomass Estimation Technique in Tallgrass Prairie

Fuel loading information is important for prescribed fire planning, evaluating wildfire risk, and understanding fire effects in grassland. Yet fuel loads in grasslands often go unmeasured because of the time required to clip plots and process samples, as well as limited access or proximity to a drying oven. We tested the digital photography biomass estimation technique for measuring fuel load in grasslands in two national parks in the eastern Great Plains. The method consists of using percentage image obstruction, as determined by digital photography, to estimate vegetation biomass based on a linear transformation (i.e., regressing dry clipped weights against percent digital obstruction). We used the technique with some modification and measured digital obstruction at two sites at Wilson’s Creek National Battlefield, Missouri (WICR), and three sites at Tallgrass Prairie National Preserve, Kansas (TAPR). The method did not result in strong correlations at either of the two sites at WICR (Site 1: r²=0.02; Site 2: r²=0.32), but performed relatively well at TAPR (Site 1 [<1 yr since burn]: r²=0.82; Site 2 [2 yr since burn]: r²=0.57; Site 3 [1 yr since burn]: r²=0.88). Linear regressions for the three sites at TAPR did not differ in slope (P>0.05). In general, the denser the vegetation, the weaker the relationship between the vegetation biomass of clip plots and the percentage image obstruction of digital images. The digital photography technique may not be useful for estimating fuel loads in grasslands with relatively high biomass (>80 g · 0.1 m^?2) or digital image obstruction >50%. Large amounts of litter may also potentially reduce the accuracy of the technique.     

RESPITE--a computer-aided guide to the prevalence of pneumonia in pig herds

An interactive computer-based guide (termed RESPITE) was developed to demonstrate to pig producers the expected level of pneumonia in pig herds which follow specified management practices known to influence the severity of pneumonia problems. A program was used to evaluate the combined effect of known risk factors present in the particular herd on the prevalence of lung lesions in slaughter pigs. The expected prevalence was estimated using formulae that were derived from observations and data in the published literature. Eleven risk factors considered are the number of pigs in the same room, all-in/all-out versus continuous flow of pigs, closed versus open herd, temperature fluctuation ratio, width of the buildings, space allotted per pig, the type of partitions separating adjacent pens, the presence or absence of diarrhoea as a clinical problem, liquid versus solid manure disposal, ascarid control efficiency, and the presence or absence of active Aujeszky's disease in the herd. Each of the factors can be assessed in the farrowing, nursery and grow-finish stages of production and the appropriate observations are entered. The computer uses the data to calculate the expected prevalence of pneumonia in animals leaving that stage of production. Of the pneumonia developing in the farrowing, nursery and grow-finish stages, 90 per cent, 75 per cent and 25 per cent respectively is considered to resolve before slaughter inspection at 100 kg. Adjusting for this degree of resolution, the program then estimates the expected final prevalence of pneumonic lesions in slaughter pigs. Comparison of predictions with slaughterhouse data from four herds indicated that the guide was realistic in its estimates.     

Surgical Removal of an Intratracheal Nodule of Ectopic Bone and Cartilage

This report describes the occurrence and subsequent removal of a nodule of bone and cartilage from the trachea of a Malamute pup. A simplified technique for resection and anastomosis of the trachea was employed. Two months postoperatively, the trachea was radiographically normal and the patient had normal exercise tolerance.     

Identification of RSVP14 and RSVP20 components by two-dimensional electrophoresis and Western-blotting.

We have already shown that RSVP14 and RSVP20, two ram seminal plasma (SP) proteins postulated to be involved in sperm capacitation and gamete interaction can protect spermatozoa against cold-shock. In this study, we use two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) for the analysis of SP proteins of Rasa Aragonesa rams, using enhanced protein solubilization in the presence of tributyl phosphine (TBP) and a polyacrylamide linear gradient gel with a narrow pH range (4-7). The image analysis of the 2D map detected 195 protein spots, with isoelectric points (pIs) ranging from 4.5 to 6.6, and molecular weight (M(r)) from 11.7 to 90.4. Staining of 2D gels with Pro-Q Emerald 300 Glycoprotein Stain revealed that most significant proteins in ram SP are glycosylated. The removing of protein N-linked oligosaccharides improved the gel resolution. 2D-PAGE analysis of the whole fraction 6 (F6) separated from ram SP by exclusion chromatography showed six main protein spots, four (a, b, c, d) in the 14 kDa and two (e, f) in the 20 kDa region. Western-blot analyses indicated that the anti-P14 antibody recognized four spots on the SP map, 4, 5, 6 and 7, that matched with spots a, b, c, d of F6 map. The anti-P20 antibody recognized spots 13 and 14 of SP map that corresponded to spots e, f of F6 map. The deduced sequences by de novo sequencing evidenced that protein spots 7 and 13 have significant similarities to BSP family, while protein spots 4 and 14 did not appear to be homologous with any reported protein in the current mammalian Proteinbank databases.