Effects of three dehorning techniques on behavior and wound healing in feedlot cattle.

Crossbred horned steers and heifers (n = 40; BW = 311.8 ± 4.7 kg) were used to determine the effect of dehorning methods on pain, cattle behavior, and wound healing. Cattle were blocked by weight and randomly assigned to 1 of 4 treatments: 1) control (CON), 2) banded using high tension elastic rubber (BAND), 3) mechanically removed (MECH), or 4) tipped (TIP). Vocalization and behavior were recorded during the dehorning process. Wound healing scores, attitude, gait and posture, appetite, and lying were recorded daily. Vocalization scores were highest for MECH cattle and BAND cattle vocalized more than TIP and CON (P < 0.05). Attitude (P = 0.06), gait and posture (P = 0.06), and lying scores (P < 0.05) were higher for BAND cattle in the days following procedures compared to MECH, TIP, and CON cattle. Cattle in the BAND treatment tended (P < 0.13) to have higher appetite scores than the other methods. Wound healing scores (horn bud and bleeding) were higher for BAND cattle than MECH, TIP, and CON cattle (P < 0.05). These data indicate that MECH is a painful procedure for cattle at the time of the procedure. Banding to remove horns from cattle is not recommended based on the data and observations from this study.

The heritability of the expression of two stress-regulated gene fragments in pigs.

Pigs reared in commercial production units sometimes encounter stressors that significantly decrease growth performance. It is hypothesized that response to stress challenges could potentially be used as selection criteria. This study aimed to investigate, in a commercial setting, the heritability of two target genes previously shown to be induced in response to stress, and related to growth performance, in an experimental situation. Blood samples (n = 2,392) were collected from three separate breeding lines of pedigreed and performance-tested boars between 24 to 25 wk of age. The expression levels of a novel fragment, '29a,' and the calcitonin receptor gene (CTR) were quantified using quantitative real-time PCR (qRT-PCR) on a subset (n = 709) of the blood samples. Gene expression levels were corrected for the efficiency of PCR reactions and also computed directly from threshold cycle (Ct) values. Resulting data showed a skewed nonnormal distribution of expression levels for the target genes relative to the endogenous control, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and were highly variable. Analyses were subsequently performed using untransformed and log-transformed data, with outliers identified and deleted in edited data sets. Regardless of the transformation or editing procedures for outliers applied, there was negligible genetic variation for the expression of target genes relative to GAPDH. In contrast, repeatabilities of replicate samples were generally high (between 0.54 and 0.67). Absolute expression levels for GAPDH and 29a were lowly heritable (h2 of about 0.04), although estimates did not exceed their SE. Subsetting the data according to whether the target gene had a higher or lower level of expression than GAPDH was then performed using the relevant Ct values. In the subset where the target gene was more highly expressed than GAPDH, a moderate estimate of heritability (0.18 +/- 0.10) for the log-transformed absolute expression level of 29a was obtained, whereas the estimate for its expression relative to GAPDH was lower (0.09 +/- 0.07). Estimates of heritability did not increase in the subset of low expression data. The limitations of using gene expression measures as potential selection criteria in commercial situations are discussed.

Effects of grouping unfamiliar cohorts, high ambient temperature and stocking density on live performance of growing pigs.

Ninety-six crossbred intact male pigs (34.5 +/- 3.5 kg BW) were allocated by weight and vocalization score to a 2 x 2 x 2 dynamic experimental design including two stocking densities (1 or 2 m(2)/pig), two temperatures (22 degrees C and 30 degrees C), and two short groupings of unfamiliar cohorts (six pigs as one pig per group, and six pigs per group). The study was conducted over 8 wk, and live weight gain (WTG) and feed intake (FI; as-fed basis) were measured weekly. During the first week, pigs were housed in individual pens from four independent rooms. To group pigs, pen partitions were removed. Pigs were grouped in Rooms 2 and 3 from wk 2 to 4, and in Rooms 1 and 4 during wk 7. Temperature was increased from 22 degrees C to 30 degrees C in Rooms 1 and 2 during wk 4 and 7. Pen partitions were replaced in Rooms 2 and 3 at the end of wk 4 and in Rooms 1 and 4 at the end of wk 7 to return pigs to their individual pens. Grouping pigs decreased FI during wk 3 (15.08 +/- 0.43 vs. 14.03 +/- 0.41 kg P < 0.10), and during wk 7 (17.42 +/- 0.46 vs. 14.24 +/- 0.41 kg; P < 0.01). In addition, grouping had a negative effect (P < 0.001) on WTG at wk 3 (7.38 +/- 0.28 vs. 5.71 +/- 0.28 kg) and at wk 7 (6.70 +/- 0.26 vs. 2.99 +/- 0.26 kg). For grouped pigs, raising the temperature decreased (P < 0.01) WTG (7.49 +/- 0.29 vs. 6.41 +/- 0.29 kg during wk 4, and 3.37 +/- 0.38 vs. 2.62 +/- 0.38 kg during wk 7). Mean FI was decreased (P < 0.01) with the 30 degrees C treatment during wk 7 only (15.49 +/- 0.33 kg at 22 degrees C compared with 12.99 +/- 0.33 kg at 30 degrees C). Compensatory feed intake was evident after the treatments had ceased at wk 6, whereby previously heat-treated grouped pigs had a higher FI (17.97 +/- 0.45 kg) than the animals individually housed at 22 degrees C (12.99 +/- 0.33 kg). Stocking density effects were noted after the grouping and high temperature treatments had ceased. For instance, during wk 5, low-density-housed pigs grew faster (P < 0.001) than their high-density counterparts (9.04 +/- 0.38 vs. 7.49 +/- 0.29 kg). In conclusion, under the conditions of this study, the grouping of unfamiliar cohorts and high ambient temperature treatments had a detrimental effect on pig performance, and these effects were reversible.

A mixed-model approach for the analysis of cDNA microarray gene expression data from extreme-performing pigs after infection with Actinobacillus pleuropneumoniae.

We proposed a novel statistical approach for the analysis of cDNA experiments based on mixed-model methodology combined with mixtures of distributions. Our objective was to detect genes that may be involved in conferring heritable differences in susceptibility to common infections in intensive pig production. We employed a microarray expression profiling strategy and a mixed-model approach to the analysis of the expression data. A cDNA microarray of pig with 6,420 probes from immune tissues and cells was used to compare gene expression in peripheral blood leukocytes of two pigs showing extreme performance in their response to infection with Actinobacillus pleuropneumoniae. Principal components analyses were used to identify the two most extreme-performing pigs after infection (i.e., pigs whose measured responses to infection fell at the extremes). Blood samples and expression profiles from 0 to 24 h after infection were compared using a bivariate, mixed-model approach, in which the effect gene x immunological status interaction was treated as a random effect. Bayesian model-based clustering via mixtures of normal distributions of the resulting BLUP of the random interaction was approached and resulted in a list of 307 differentially expressed genes, of which 179 were down-regulated in the susceptible pig. The majority of the differentially expressed genes were derived from a cDNA library of leukocytes of A. pleuropneumoniae-challenged pigs that were subtracted against leukocytes before the challenge. These results provide evidence that the proposed statistical approach was useful in enhancing the knowledge of the mechanisms involved in the genetics of the immune response.

Antisense oligonucleotide blockade of connexin expression during embryonic bone formation: evidence of functional compensation within a multigene family.

Prior studies in our laboratory demonstrated the presence of gap junction proteins (connexins) throughout intramembranous bone formation [Minkoff et al. (1994) Anat Embryol 190:231-241]. In addition, two members of the connexin family of gap junction proteins, connexin 43 (Cx43; Gj alpha 1) and connexin 45 (Cx45; Gj alpha 6), were found by Civitelli et al. [1993; J Clin Invest 91:1888-1896] to be associated, specifically, with osteogenesis. Recently, however, a null mutation in the gene encoding Gj alpha 1 in mice has been produced by Reaume et al. [1995; Science 267:1831-1834]. Gj alpha 1 null homozygotes survived to term but died at birth of heart abnormalities. Examination of the null homozygous embryos, surprisingly, did not reveal overt histological or anatomical abnormalities in any organ system other than the heart. In view of this, the present investigation was initiated in order to evaluate bone formation under conditions in which the expression of Gj alpha 1 and Gj alpha 6, the connexins specifically associated with osteogenesis, had been perturbed, individually as well as in combination. An in vitro system employing organ cultures of dissociated embryonic chick mandibular mesenchyme was employed. Mesenchyme was cultured in the presence and absence of sense and antisense oligodeoxynucleotides (ODN), ranging in length from 15 to 24 mer and containing sequences that included the initiation codon of Gj alpha 1 and of Gj alpha 6. In cultures of mesenchyme, grown for 6 to 13 days in the presence of the combined antisense ODNs to Gj alpha 1 and Gj alpha 6, bone formation was markedly reduced or absent. By contrast, in cultures grown in medium containing the combination of corresponding sense ODNs to both Gj alpha 1 and Gj alpha 6, bone formation was evident. In addition, when cultures were grown in the presence of antisense or sense ODNs to either Gj alpha 1 or Gj alpha 6, individually, bone formation was seen. Immunohistochemical analysis of connexin expression revealed intense immunoreactive signal to Gj alpha 1 and Gj alpha 6 in bone of the control explants, in which no ODNs were present; in those cultures in which either Gj alpha 1 and Gj alpha 6 antisense ODNs were present, however, the expression of the respective connexin protein was either significantly reduced or absent. Further, in those explants in which Gj alpha 1 expression was blocked, immunoreactive signal to Gj alpha 6 appeared to have been amplified in regions of developing bone. These results suggest that, in avian osteogenic tissue, when Gj alpha 1 protein expression has been impeded another related connexin protein (Gj alpha 6) may subserve the functions of the missing connexin. The findings of this study, therefore, support the hypothesis that, within the connexin gene family, functional compensation can occur.

Evaluation of enzymatically modified potato starches in diets for weanling pigs.

We conducted three growth trials to evaluate replacing carbohydrate sources with enzymatically modified potato starches in diets for weanling pigs. In Exp. 1, 180 pigs (initially 5.3 kg and 21+/-2 d of age) were used to compare the effects of corn (36.5%), edible-grade oat flour (36.5%), two enzymatically modified potato starches (12%), and added lactose (12%) on pig performance. Potato Starch 1 had a dextrose equivalent (DE) of 6 and Potato Starch 2 had a DE of 20; both were spray-dried maltodextrans. Pigs that were fed Potato Starch 2 had greater (P<.05) ADG and ADFI than pigs fed diets that contained corn or oat flour from d 0 to 14 after weaning, and pigs fed either Potato Starch 1 or added lactose had intermediate ADG and ADFI. However, for the overall trial (d 0 to 35), no differences (P>.10) in growth performance were observed. In Exp. 2, 198 pigs (initially 4.3 kg and 19+/-2 d of age) were used to determine whether modified Potato Starch 2 could replace a portion of the corn or lactose in the diet. The control diet contained 10% dried whey, and additional treatments were formulated by adding 7 or 14% modified Potato Starch 2 or lactose in place of corn. A positive control diet was formulated containing 29% dried whey. From d 0 to 14 after weaning, increasing dietary lactose improved (linear, P<.04) ADG and ADFI. Increasing the potato starch had no effect on ADG but increased ADFI (linear, P<.02). In Exp. 3, 180 pigs (initially 3.9 kg and 14+/-3 d of age) were used to evaluate Potato Starch 2 or 3 (DE = 30, a spray-dried glucose syrup) as replacements for either corn or lactose in the diet. Pigs were fed a control diet containing 15% dried whey and 12% added lactose. Twelve percent modified Potato Starch 2 or 3 replaced either corn or lactose in the diet on a wt/wt basis. From d 0 to 14 and d 0 to 21, pigs fed either modified potato starch substituted for corn had greater (P<.07) ADG than those fed the control diet. Pigs fed diets with either modified starch substituted for lactose had similar ADG as those fed the control diet. These results suggest that when substituted for corn, Potato Starch 2 can improve growth performance of weanling pigs.

Evidence of multiple mechanisms of avermectin resistance in haemonchus contortus--comparison of selection protocols.

Three isolates of Haemonchus contortus selected for avermectin resistance in sheep were compared in three in vitro pharmacological tests previously shown to discriminate between field isolates of H. contortus resistant and susceptible to the avermectins. Two isolates, F7-A and IVC, were selected for avermectin resistance in the laboratory from a reference susceptible isolate using suboptimal doses of ivermectin (LD95) for 7 and 16 generations, respectively. In these isolates avermectin resistance was not associated with a decreased sensitivity to avermectin inhibition of larval development or L3 motility but was associated with an increased sensitivity to paraherquamide. The third isolate, Warren, was derived from an overwhelmingly avermectin-susceptible, mixed species field isolate in a single generation by propagating the small number of survivors of a 0.2 mg/kg ivermectin treatment (i.e. 10 x LD95). This isolate, like previously characterised avermectin-resistant H. contortus isolates derived from the field in South Africa and Australia, showed a markedly reduced sensitivity to avermectin inhibition of larval development and L3 motility, as well as an increased sensitivity to paraherquamide. These results suggest that avermectin resistance can manifest itself in different ways and that the two selection protocols used to generate the F7-A, IVC and Warren isolates have resulted in the selection of different resistance phenotypes.

Evaluation of potato proteins on the growth performance of early-weaned pigs.

We conducted five experiments to evaluate conventional and low-glycoalkaloid potato protein (CPP and LGPP, respectively) in diets for early-weaned pigs. In Exp. 1, 150 weanling pigs (initially 4.4 +/- .9 kg and 15.5 +/- 2 d of age) were fed either a control diet containing 3% spray-dried animal plasma (SDAP) or diets with additional SDAP (2.5 or 5% added; 5.5 or 8% total) or CPP (2.6% or 5.1%) substituted on a total lysine basis. From d 0 to 14 after weaning, increasing SDAP increased (linear, P < .05) ADG and ADFI, whereas increasing CCP had no effect on growth performance. In Exp. 2, 180 weanling pigs (initially 5.9 +/- 1.2 kg and 20 +/- 2 d of age) were fed diets containing a LGPP replacing 25, 50, 75, or 100% of the 7% dietary SDAP on a digestible lysine basis. From d 0 to 7 after weaning, increasing LGPP increased and then returned to control levels ADG and ADFI (quadratic, P < .01) and gain:feed ratio (quadratic, P < .05). In Exp. 3, 175 weanling pigs (initially 5.5 +/- 1.1 kg and 20 +/- 3 d of age) were fed either a control diet containing 20% dried whey, 17.5% dried skim milk, and 4% select menhaden fish meal (SMFM) or diets consisting of lactose and either 3.5 and 7.0% SDAP or 4.0 and 8.0% LGPP added at the expense of dried skim milk on a digestible lysine basis. From d 0 to 7 after weaning, ADG and ADFI increased (linear, P < .05) with increasing SDAP. With increasing LGPP, ADG and ADFI increased and then decreased (quadratic, P < .10 and P < .05, respectively). Gain:feed ratio (G/F) was not affected by SDAP and was improved (linear, P < .05) for pigs fed increasing LGPP. In Exp. 4, 270 weanling pigs (initially 6.2 +/- 1.6 kg and 20 +/- 3 d of age) were used to compare three diets that contained either 2.5% spray-dried blood meal (SDBM), 4.8% SMFM, or 3.92% CPP; test feedstuffs were substituted on a total lysine basis and diets were fed from d 7 to 28 after weaning. Pigs fed CPP had decreased (P < .05) ADG and G/F compared with those fed the other protein sources. In Exp. 5, 255 weanling pigs (initially 5.3 +/- 1.2 kg and 17 +/- 2 d of age), were used to compare five diets that contained either 2.5% SDBM, 5.51% SMFM, 4.17% CPP, 4.17% LGPP or 8.34% LGPP; feedstuffs were substituted on a digestible lysine basis and diets were fed from d 7 to 28 after weaning. No differences (P > .10) were observed in growth performance among pigs fed any of the protein sources within the experiment. However, pigs fed the LGPP had numerically greater ADG and better G/F than those fed CPP. In conclusion, these results suggest that LGPP can be an effective replacement for a portion of the SDAP in diets for weanling pigs.

Optimization of targeted RNA recombination and mapping of a novel nucleocapsid gene mutation in the coronavirus mouse hepatitis virus.

We have recently described a method of introducing site-specific mutations into the genome of the coronavirus mouse hepatitis virus (MHV) by RNA recombination between cotransfected genomic RNA and a synthetic subgenomic mRNA (C. A. Koetzner, M. M. Parker, C. S. Ricard, L. S. Sturman, and P. S. Masters, J. Virol. 66:1841-1848, 1992). By using a thermolabile N protein mutant of MHV (Alb4) as the recipient virus and synthetic RNA7 (the mRNA for the nucleocapsid protein N) as the donor, we selected engineered recombinant viruses as heat-stable progeny resulting from cotransfection. We have now been able to greatly increase the efficiency of targeted recombination in this process by using a synthetic defective interfering (DI) RNA in place of RNA7. The frequency of recombination is sufficiently high that, with Alb4 as the recipient, recombinants can be directly identified without using thermal selection. The synthetic DI RNA has been used to demonstrate that the lesion in another temperature-sensitive and thermolabile MHV mutant, Alb1, maps to the N gene. Sequencing of the Alb1 N gene revealed two closely linked point mutations that fall in a region of the N molecule previously noted as being the most highly conserved region among all of the coronavirus N proteins. Analysis of revertants of the Alb1 mutant revealed that one of the two mutations is critical for the temperature-sensitive phenotype; the second mutation is phenotypically silent.

Populations, practices, and problems in forensic psychiatric facilities.

This is a study of the public facilities to which mentally disordered offenders are committed or transferred so that they may be securely confined while simultaneously participating in programs designed for treatment of their mental disorders. The study focuses principally on the nature and characteristics of these facilities: their patient populations, staff, security conditions, treatment programs, and operational programs. We identified and surveyed 231 facilities. The information from the survey, legal research, and site visits to 11 programs has been integrated and used to address four major issues: the types of facilities mentally disordered offenders are institutionalized in for treatment of their mental disorders; the legal, diagnostic, and demographic characteristics of the residents of these facilities; the forms of treatment and levels of staffing available in these facilities; and the common problems faced by facility administrators with respect to facility management, treatment, and release decisions.