The effect of ovine peripheral blood mononuclear cells on Haemonchus contortus larval morbidity in vitro.

The objective of this study was to determine the effects of peripheral blood mononuclear cells (PBMC), derived from parasite-resistant St. Croix (STC) hair sheep and parasite-susceptible Suffolk (SUF) sheep, on Haemonchus contortus L3 stage larval death in vitro, with or without autologous serum. Larval morbidity was quantified by measuring larval ATP concentration following incubation with PBMC. Larvae exposed to either STC- or SUF-derived PBMC had lower ATP than live larvae (0.12 µmol/L ATP and 0.16 µmol/L ATP vs 0.27 µmol/L ATP, respectively) (P<.001) and greater ATP of dead larvae (0.03 µmol/L ATP) (P<.001). Breed differences were observed with addition of autologous serum. Larvae exposed to SUF-derived PBMC with autologous serum were not significantly different from live larval ATP. STC-derived serum did not significantly reduce larval ATP compared to PBMC alone (0.11 µmol/L ATP), but was significantly reduced compared to live larvae (0.22 µmol/L ATP) and SUF-derived PBMC with autologous serum (0.23 µmol/L ATP) (P<.001). These data indicate that a cellular response alone is capable of significantly reducing larval ATP in a breed-independent manner. However, addition of serum to SUF-PBMC failed to reduce larval ATP, indicating breed-dependent humoral response to H. contortus.

Serum-mediated Haemonchus contortus larval aggregation differs by larval stage and is enhanced by complement.

The objectives of this study were to measure Haemonchus contortus larval aggregation by complement/antibody complexes, determine effect of breed resistance and infection status and determine the effect of larval maturation on larval aggregation in vitro. Larval binding assays were performed on H. contortus L3, exsheathed L3 and L4 incubated with serum from either parasite naïve or H. contortus primed St. Croix (resistant) and Suffolk (susceptible) lambs. No differences in L3 aggregation were observed between serum from either breed or infection status. Exsheathed L3 (60%) and L4 (42%) aggregation by primed Suffolk serum was significantly reduced compared with L3 (80%, P<.001). Removal of either complement or antibody effectively eliminated L3 aggregation (P<.001). Combination of antibody-depleted and complement-inactivated serum restored L3 aggregation to levels consistent with unprocessed serum, supporting a role for antibody and complement in aggregation (P<.001). Use of fluorescence-labelled anti-sheep IgG antibody allowed documentation of IgG bound to serum complexes within L3 masses and was present only in larvae incubated with normal serum, and complement- and antibody-depleted serum combination. These data indicate that complement/antibody complexes inhibit larval motility through larval aggregation which may be critical in early larval clearance of H. contortus.

Effect of early grain feeding of beef steers on postabsorptive metabolism.

The objective of this study was to determine the effect of early weaning followed by a period of high-grain feeding on plasma acetate kinetics and signaling protein phosphorylation in LM tissue of growing steers. We hypothesized that early grain feeding would result in altered cell signaling and acetate use to support observed improvements in carcass gain and marbling. Fall-born Angus × Simmental steers were weaned at 106 ± 4 d of age (early weaned [EW]; n = 6) and fed a high-grain diet for 148 d or remained with their dams (normal weaned [NW]; n = 6) on pasture until weaning at 251 ± 5 d of age. Both treatments were subsequently combined and grazed on mixed summer pasture to 394 ± 5 d of age followed by a feedlot ration until harvest at 513 ± 5 d of age. Longissimus muscle tissue biopsies were collected at 253 ± 5 and 394 ± 5 d of age and at harvest. Total and phosphorylated forms of 5' adenosine monophosphate-activated protein kinase (AMPK) and downstream proteins of the mammalian target of rapamycin signaling pathway were determined by western blotting. Eight steers were used to assess acetate clearance at different age points via a bolus infusion of acetate (4 mmol/kg of BW). Early weaned steers had greater (P < 0.05) ADG than NW steers during the early grain feeding period. Phosphorylated to total ratios of ribosomal protein S6 (rpS6) and ribosomal protein S6 kinase 1 (S6K1) were significantly different during the early grain feeding period. Phosphorylated to total ratios of S6K1, rpS6, acetyl-CoA carboxylase, and 4E binding protein 1 and the absolute amount of phosphorylated AMPK were correlated with ADG, explaining 46% of the variance. Acetate clearance rates were less (P < 0.05) and synthesis rates were greater (P = 0.06) in EW steers during early grain feeding. Acetate synthesis rates were also greater (P < 0.05) in NW steers at harvest, suggesting a permanent shift in the gut microflora or gut function in response to the treatment. Neither treatment nor acetate infusion significantly affected plasma glucose or insulin concentrations. Plasma ß-hydroxybutyric acid concentrations increased with acetate infusion (P < 0.05). Based on these results, altered cell signaling during the early grain feeding period likely mediated increased protein deposition, leading to increased carcass weights, but observed changes in acetate appearance and clearance rates do not appear to explain the observed differences in intramuscular fat deposition during the terminal feeding period.

Early metabolic imprinting events increase marbling scores in fed cattle.

Early weaning of calves to a high concentrate diet results in greater fat deposition and suggests early postnatal metabolic imprinting events may be exploited as a management tool to improve cattle value. Our objective was to implement a short, high energy dietary intervention before a typical grazing period to manipulate intramuscular fat deposition in finishing cattle. Fall-born, Angus-sired steer calves (n = 24) were stratified by sire and randomly assigned to normal weaned (NW) or metabolic-imprinted (MIP) treatments. At 105 ± 6d (135kg), MIP calves were transitioned to a diet containing 20% CP and 1.26 Mcal/kg NEg. Metabolic-imprinted calves were fed ad libitum as a group. Normal weaned calves remained on their dam until 253 ± 6 d of age. At this time, treatment groups were combined and grazed for 156 d on a mixed summer pasture. Following the grazing phase, steers were adapted to a corn silage-based feedlot diet and performance was monitored on 28-d intervals. Calves were staged for harvest based on backfat endpoint (target 1.0 to 1.2 cm). Metabolic-imprinted calves were heavier (P < 0.05) than NW calves (341 vs. 265 ± 4.2 kg) at normal weaning age. During the grazing phase, NW steers gained more weight than (P < 0.05) MIP steers (0.69 vs. 0.35 ± 0.03 kg/d). Feedlot performance and USDA yield grade were similar (P > 0.20) between treatments. However, MIP steers produced heavier (P < 0.05) carcasses (564 vs. 524 ± 5.6 kg) with higher (P < 0.001) marbling scores (645 vs. 517 ± 23). Therefore, calves consuming a high concentrate diet for 148 d after early weaning produced higher quality carcasses. This suggests early weaning and feeding a high concentrate before grazing is a viable strategy to increase marbling deposition compared with a traditional production system.

Validation of live animal ultrasonic measurements of body composition in market lambs.

Market lambs from the state fair of Virginia (n = 172) were ultrasonically evaluated by 4 scan technicians and 3 image interpreters to determine accuracy of ultrasonic estimates of loin muscle area (ULMA), backfat thickness (UBF), and body wall thickness (UBW). Lambs were initially scanned at the preferred magnification setting of each technician; 2 chose 1.5x and 2 chose 2.0x. Lambs were then scanned a second time for ULMA and UBF with machine magnification settings changed from 1.5 to 2.0x, or vice versa, midway through the second scan. Lambs were then slaughtered, and analogous measurements [carcass loin muscle area, carcass backfat thickness, and carcass body wall thickness (CBW)] were recorded on chilled carcasses. Pooled, residual correlation coefficients within technicians and interpreters between ultrasonic measurements from the first scan and carcass measurements were 0.66 for loin muscle area, 0.78 for backfat thickness, and 0.73 for body wall thickness, but were reduced to 0.43, 0.69, and 0.50, respectively, by inclusion of linear effects of carcass weight in the model. Mean bias for technicians and interpreters ranged from -1.30 to -2.66 cm(2) for loin muscle area, -0.12 to -0.17 cm for backfat thickness, and 0.14 to -0.03 cm for body wall thickness; prediction errors ranged from 1.86 to 2.22 cm(2), 0.12 to 0.14 cm, and 0.35 to 0.38 cm, respectively. Pooled correlations between repeated measures were 0.67 for ULMA, 0.79 for UBF, and 0.68 for UBW at the same magnification and 0.73 for ULMA and 0.76 for UBF across different magnification settings. Mean differences between repeated measures were more variable among technicians and interpreters than statistics comparing ultrasound to carcass measures. Standard errors of repeatability ranged from 1.61 to 2.45 cm(2) for ULMA, 0.07 to 0.11 cm for UBF, and 0.36 to 0.42 cm for UBW. The effect of changing magnification setting on technician and interpreter repeatability was small for UBF and ULMA. The accuracy of prediction of CBW from UBW was similar to that achieved for backfat thickness; further assessment of the value of ultrasonic measurements of body wall thickness in lambs is warranted. These results indicate that ultrasound scanning can reliably predict carcass loin muscle area and backfat thickness in live lambs and, accordingly, has value in selection programs to improve composition. Development of certification standards for US lamb ultrasound technicians based on results of this study and others is proposed.

Longitudinal changes in ultrasonic measurements of body composition during growth in Suffolk ram lambs and evaluation of alternative adjustment strategies for ultrasonic scan data.

Four equations were used to compare alternative procedures to adjust ultrasonic estimates (y) of backfat thickness (BF) and LM area (LMA) for BW using data from a series of 7 scans on 24 Suffolk ram lambs born in 2007. Equations were linear, linear + quadratic, allometric (y = alphaBW(beta)), and allometric + BW (ABW; y = alphaBW(beta)e(gammaW)). Goodness of fit was very similar between equations over the range of the data. Resulting adjustment equations were tested using 3 serial scans on winter-born Suffolk (n = 150), Hampshire (n = 36), and Dorset (n = 43) rams and 52 fall-born Dorset rams tested at the Virginia Ram Test in 1999 through 2002. Partial correlations (accounting for the effect of year) between predicted and actual measures ranged from 0.78 to 0.87 for BF and 0.66 to 0.93 for LMA in winter-born rams and from 0.70 to 0.71 for BF and 0.72 to 0.78 for LMA in fall-born rams. No significant differences in predictive ability existed between equations for BF or LMA (P > 0.05), and there was no indication that the allometric equation was a better predictor than linear within the range of the data. Adjustment equations were also tested using serial scan data from 37 Suffolk ewe lambs born in the same contemporary group as the rams used to derive the prediction equations but fed for a substantially slower rate of BW gain. Correlations between predicted and actual values of BF and LMA indicated lambs were too young and small at the first scan (77 d, 32.4 kg) to reliably predict carcass measures at typical slaughter weights. For prediction using data from the 2 subsequent scans, at mean ages >96 d and mean BW >39 kg, correlations between predicted and actual values were 0.72 to 0.74 for BF and 0.54 to 0.76 for LMA. Little difference existed between equations for predicting BF. For LMA, the ABW form was a weaker predictor than the others, and the linear equation was slightly superior to allometric. Therefore, it appears the linear and allometric forms are both suitable for use in central ram test and performance-tested farm flocks.

Growth and carcass characteristics of lambs sired by Dorper and Dorset rams.

Growth and carcass merit of Dorset-(DO) and Dorper-sired (DP) lambs were compared over 3 yr in matings with 50% Dorset, 25% Rambouillet, 25% Finnsheep ewes. The DP were slightly lighter (P = 0.09) at birth than the DO lambs. In the first year of the study, DP lambs produced by AI using imported South African sires were heavier than DO lambs when weaned at 60 d of age (21.7 vs. 19.5 kg; P = 0.05). In yr 2 and 3, however, offspring of natural-service Dorper sires produced in the U.S. did not differ in weaning weight from DO lambs (16.9 vs. 17.8 kg; P = 0.02 for breed x year interaction). Lamb survival was also affected by breed x year interaction (P = 0.04). In 2000 and 2001, with 12 to 16% triplet or larger litters, mortality was higher for DP lambs (14.9 vs. 7.7%; P = 0.12). However, in 2002, with approximately 33% triplet or larger litters and with higher mortality levels in all birth types, DP lambs had fewer death losses than did DO lambs (23.2 vs. 36.1%; P = 0.11). No differences between DO and DP lambs were observed in postweaning gain during summer grazing or in drylot in autumn. At chilled carcass weights of approximately 25 kg, DP lambs were somewhat fatter than DO lambs, with greater body wall thickness (P < 0.01; 22 vs. 19 mm) and slightly greater backfat thickness (P = 0.15; 6.4 vs. 5.5 mm) and yield grades (P = 0.15; 2.9 vs. 2.6). The DP lambs also had more desirable leg scores (P = 0.01; 11.6 vs. 10.9) and slightly larger LM area (P = 0.13; 14.1 vs. 13.5 mm2) than did DO lambs, confirming acceptable muscling and conformation in carcasses from Dorper-sired lambs. However, differences were not observed in the percentage of carcass weight in the leg or loin, or in the lean:bone ratio in the dissected leg. Ultrasonic measurements of backfat thickness and LM area taken in live lambs before slaughter were positively associated with direct measures on chilled carcasses with correlations of 0.77 for backfat thickness and 0.51 for LM area.

Performance of hair sheep composite breeds: resistance of lambs to Haemonchus contortus.

This study was designed to evaluate breed differences in resistance to Haemonchus contortus in lambs. A total of 181 ewe lambs representing crossbred Dorsets (DO) and Dorpers (DP; out of 1/2-Dorset, 1/4-Rambouillet, 1/4-Finnsheep ewes) and straight-bred Katahdins (KT) were evaluated over 3 yr. An additional 144 DO, DP, KT, and Barbados Blackbelly x St. Croix (HH) wethers were evaluated over 2 yr. Lambs were weaned at 60 to 90 d of age. After deworming at about 4 mo of age, ewe lambs received approximately 10,000 infective larvae and were evaluated for parasite resistance in drylot, whereas wethers were evaluated on pasture under conditions of natural infection. Each sex was analyzed separately. Egg counts per gram of feces (FEC), log-transformed FEC (LFEC), packed cell volumes (%), and body weights (kg) measured at 3, 4, 5, and 6 wk after deworming and reinfection were analyzed. Breed influenced all traits (P < 0.05) except BW in ewe lambs and PCV in wethers. Year and week influenced (P < 0.05) all traits. At most times, DP had the highest FEC, DO had the lowest PCV, and KT and HH had lower FEC and higher PCV than either DO or DP. Clearly, Dorper sheep were not more resistant to parasites than DO, but they were able to cope with infection better by maintaining similar or higher (P < 0.05) PCV and similar BW. Katahdin and HH were more resistant, with lower FEC (P < 0.05) than DO or DP. Breed differences were more apparent when infection levels were higher; DO and DP were less affected when infection levels were low and when animals were on a better plane of nutrition. Caribbean hair sheep originated in hot, humid regions of West Africa. They and the derivative Katahdin breed seem relatively resistant to parasitism. The Dorper, in contrast, was developed in more arid regions and exhibited little parasite resistance.

Prediction of retail product weight and percentage using ultrasound and carcass measurements in beef cattle.

Data from 534 steers representing six sire breed groups were used to develop live animal ultrasound prediction equations for weight and percentage of retail product. Steers were ultrasonically measured for 12th-rib fat thickness (UFAT), rump fat thickness (URPFAT), longissimus muscle area (ULMA), and body wall thickness (UBDWALL) within 5 d before slaughter. Carcass measurements included in USDA yield grade (YG) and quality grade calculations were obtained. Carcasses were fabricated into boneless, totally trimmed retail products. Regression equations to predict weight and percentage of retail product were developed using either live animal or carcass traits as independent variables. Most of the variation in weight of retail product was accounted for by live weight (FWT) and carcass weight with R2 values of 0.66 and 0.69, respectively. Fat measurements accounted for the largest portion of the variation in percentage of retail product when used as single predictors (R2 = 0.54, 0.44, 0.23, and 0.54 for UFAT, URPFAT, UBDWALL, and carcass fat, respectively). Final models (P < 0.10) using live animal variables included FWT, UFAT, ULMA, and URPFAT for retail product weight (R2 = 0.84) and UFAT, URPFAT, ULMA, UBDWALL, and FWT for retail product percentage (R2 = 0.61). Comparatively, equations using YG variables resulted in R2 values of 0.86 and 0.65 for weight and percentage of retail product, respectively. Results indicate that live animal equations using ultrasound measurements are similar in accuracy to carcass measurements for predicting beef carcass composition, and alternative ultrasound measurements of rump fat and body wall thickness enhance the predictive capability of live animal-based equations for retail yield.

Accuracy of predicting weight and percentage of beef carcass retail product using ultrasound and live animal measures.

Five hundred thirty-four steers were evaluated over a 2-yr period to develop and validate prediction equations for estimating carcass composition from live animal ultrasound measurements and to compare these equations with those developed from carcass measurements. Within 5 d before slaughter, steers were ultrasonically measured for 12th-rib fat thickness (UFAT), longissimus area (ULMA), rump fat thickness (URPFAT), and body wall thickness (UBDWALL). Carcasses were fabricated to determine weight (KGRPRD) and percentage (PRPRD) of boneless, totally trimmed retail product. Data from steers born in Year 1 (n = 282) were used to develop prediction equations using stepwise regression. Final models using live animal variables included live weight (FWT), UFAT, ULMA, and URPFAT for KGRPRD (R2 = 0.83) and UFAT, URPFAT, ULMA, FWT, and UBDWALL for PRPRD (R2 = 0.67). Equations developed from USDA yield grade variables resulted in R2 values of 0.87 and 0.68 for KGRPRD and PRPRD, respectively. When these equations were applied to steers born in Year 2 (n = 252), correlations between values predicted from live animal models and actual carcass values were 0.92 for KGRPRD, and ranged from 0.73 to 0.76 for PRPRD. Similar correlations were found for equations developed from carcass measures (r = 0.94 for KGRPRD and 0.81 for PRPRD). Both live animal and carcass equations overestimated (P < 0.01) actual KGRPRD and PRPRD. Regression of actual values on predicted values revealed a similar fit for equations developed from live animal and carcass measures. Results indicate that composition prediction equations developed from live animal and ultrasound measurements can be useful to estimate carcass composition.

The relationship between ultrasound measurements and carcass fat thickness and longissimus muscle area in beef cattle.

Five hundred thirty-four steers were evaluated over a 2-yr period to determine the accuracy of ultrasonic estimates of carcass 12th-rib fat thickness (CFAT) and longissimus muscle area (CLMA). Within 5 d before slaughter, steers were ultrasonically measured for 12th-rib fat thickness (UFAT) and longissimus muscle area (ULMA) using an Aloka 500V real-time ultrasound machine equipped with a 17.2-cm, 3.5-MHz linear transducer. Overall, correlation coefficients between ultrasound and carcass fat and longissimus muscle area were 0.89 and 0.86, respectively. Correlations for UFAT with CFAT were similar between years (0.86 and 0.90), whereas the relationship between ULMA and CLMA was stronger in yr 1 (r = 0.91; n = 282) than in yr 2 (r = 0.79; n = 252). Differences between ultrasonic and carcass measurements were expressed on both an actual (FDIFF and RDIFF) and absolute (FDEV and RDEV) basis. Mean FDIFF and RDIFF indicated that ultrasound underestimated CFAT by 0.06 cm and overestimated CLMA by 0.71 cm2 across both years. Overall mean FDEV and RDEV, which are indications of the average error rate, were 0.16 cm and 3.39 cm2, respectively. Analysis of year effects revealed that FDIFF, FDEV, and RDEV were greater (P < 0.01) in magnitude in yr 1. Further analysis of FDEV indicated that leaner (CFAT < 0.51 cm) cattle were overestimated and that fatter (CFAT > 1.02 cm) cattle were underestimated with ultrasound. Similarly, steers with small CLMA (< 71.0 cm2) were overestimated, and steers with large CLMA (> 90.3 cm2) were underestimated. The thickness of CFAT had an effect (P < 0.05) on the error of UFAT and ULMA measurements, with leaner animals being more accurately evaluated for both traits. Standard errors of prediction (SEP) adjusted for bias of ultrasound measurements were 0.20 cm and 4.49 cm2 for UFAT and ULMA, respectively. Differences in SEP were observed for ULMA, but not UFAT, by year. These results indicate that ultrasound can be an accurate estimator of carcass traits in live cattle when measurements are taken by an experienced, well-trained technician, with only small differences in accuracy between years.

Model for the porphyrin-DNA binding site: ENDOR investigations of Cu-porphyrins binding to DNA.

Proton ENDOR has been observed from frozen solutions (ca. 38K degrees) of copper meso-(4-N-tetra-methylpyridyl)porphyrin (CuTMpyP(4)) complexed with Salmon sperm DNA in water and D2O. Lines from exchangeable protons of the DNA bases have been observed in these ENDOR spectra. Analyses of these ENDOR data show that the separations of these DNA protons from the copper atom are between 3.76 and 3.84 A with angles of 19.5 to 22.5 degrees between the Cu-H vectors and the gz axis. A distant ENDOR response has also been observed from phosphorous nuclei in the DNA backbone. We estimate that the phosphorous atoms producing this ENDOR signal are 7.5-10 A from the copper center of the porphyrin. These ENDOR data combined with results from an earlier NMR investigation have been used to construct a computer simulated model of the binding site in which the porphyrin is partially intercalated and extends into the major groove of DNA. The two GC base pairs at this site are slightly inequivalent. For each, the G imino proton and one of the C amino protons are at appropriate positions to account for the ENDOR signals arising from exchangeable protons. It is unlikely that this inequivalence would persist at room temperature where dynamic processes would give an apparently symmetric interaction. Although the model accounts for all reported experimental data involving tetracationic porphyrin species which have been suggested to be intercalators, it is not a unique solution.