Pathway-Based Approaches for Assessing Biological Hazards of Complex Mixtures of Contaminants: A Case Study in the Maumee River.

Assessment of ecological risks of chemicals in the field usually involves complex mixtures of known and unknown compounds. We describe the use of pathway-based chemical and biological approaches to assess the risk of chemical mixtures in the Maumee River (OH, USA), which receives a variety of agricultural and urban inputs. Fathead minnows (Pimephales promelas) were deployed in cages for 4 d at a gradient of sites along the river and adjoining tributaries in 2012 and during 2 periods (April and June) in 2016, in conjunction with an automated system to collect composite water samples. More than 100 industrial chemicals, pharmaceuticals, and pesticides were detected in water at some of the study sites, with the greatest number typically found near domestic wastewater treatment plants. In 2016, there was an increase in concentrations of several herbicides from April to June at upstream agricultural sites. A comparison of chemical concentrations in site water with single chemical data from vitro high-throughput screening (HTS) assays suggested the potential for perturbation of multiple biological pathways, including several associated with induction or inhibition of different cytochrome P450 (CYP) isozymes. This was consistent with direct effects of water extracts in an HTS assay and induction of hepatic CYPs in caged fish. Targeted in vitro assays and measurements in the caged fish suggested minimal effects on endocrine function (e.g., estrogenicity). A nontargeted mass spectroscopy-based analysis suggested that hepatic endogenous metabolite profiles in caged fish covaried strongly with the occurrence of pesticides and pesticide degradates. These studies demonstrate the application of an integrated suite of measurements to help understand the effects of complex chemical mixtures in the field. Environ Toxicol Chem 2021;40:1098-1122. © 2020 SETAC. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

The effect of ractopamine hydrochloride (Paylean(®)) on lean carcass yields and pork quality characteristics of heavy pigs fed normal and amino acid fortified diets.

Effects of ractopamine hydrochloride (RAC) on carcass parameters in heavy weight (133.24±8.07kg) finishing pigs (n=278) given amino acid fortified (AA) or 16% crude protein (CP) diets were evaluated. A total of seven experimental diets were formulated; RAC was added at 0, 5 and 20ppm to the 16% CP diets (CP0, CP5 and CP20, respectively) and at 0, 5, 10 and 20ppm to the AA fortified diets (AA0, AA5, AA10 and AA20, respectively). Carcass, tenderloin, and ham weights were heavier (P<0.05) for RAC AA diets vs. AA0. Loin weight was heavier (P<0.05) for AA20 vs. AA0 and CP20 vs. CP0. No differences (P>0.05) were observed for color or firmness scores. Carcass muscle score, ham weight and protein% were greater (P<0.05) for RAC diets. Moisture was greater (P<0.05) and fat was lower (P<0.05) for AA5 and AA20 vs. AA0 and CP5 and CP20 vs. CP0. Feeding RAC to late finishing swine increases carcass yields and protein% with lower fat% for pigs weighing up to 136kg.

Behavior of immunologically castrated barrows in comparison to gilts, physically castrated barrows, and intact male pigs.

The behavior of immunologically castrated barrows (IC; with Improvest; Zoetis, Parsippany, NJ) was compared with that of intact males (IM), physically castrated barrows (PC), and gilts (G). The study used 160 commercial crossbred pigs in a randomized complete block design (blocking factor was start date of study) and was performed over an 8-wk period from 16 wk of age (67.2 ± 2.50 kg BW) to a final BW of 126.5 ± 6.05 kg. The first Improvest dose was given to IC at the start of the study (d 0), and the second dose was given 28 d later. Pigs were housed in single-gender groups of 4 and had ad libitum access to feed and water. General, aggressive, and sexual behaviors were observed over a 12-h period from 0600 to 1800 h by the same trained investigators on d -1 (d prior to first dose), 13, 27 (day prior to second dose), 34, 41, and 55 of study (end of test). General behaviors (number of pigs lying, sitting, standing, at feeder, and at drinker) were recorded every 10 min in all pens (10 pens/gender), and aggressive (bites, head butts, and fights) and sexual (mounts) behaviors were recorded continuously over the 12-h period on a subsample of 4 pens/gender. There was no difference ( > 0.05) between the genders for lying, sitting, or drinker-related behavior. For the 4-wk period prior to second dose, a greater percentage of PC and G were at the feeder ( < 0.05) than IC or IM (8.0%, 7.4%, 10.2%, and 9.3% for IC, IM, PC, and G, respectively; SEM = 0.44). However, after the second dose, the percentage of pigs at the feeder was similar ( > 0.05) for IC, PC, and G and was greater ( < 0.05) for those genders than IM (10.1%, 7.1%, 10.0%, and 8.8%, respectively; SEM = 0.50). Prior to the second dose, the incidence of aggressive behaviors was generally greater for IC and IM compared with PC and G; however, after the second dose, the incidence of aggressive behaviors was similar for IC and PC and lower ( < 0.05) for those genders than for IM. The frequency of mounts for IC was greater ( < 0.05) than for PC before (25.5, 27.3, 2.5, and 1.5 total mounts/pen, respectively; SEM = 4.37) but not after ( > 0.05; 3.3, 33.3, 0.25, and 0.25 total mounts/pen, respectively; SEM = 1.15) the second dose. These results suggest that prior to the second Improvest dose, the behavior of IC was similar to that of IM and transitioned to become more similar to the behavior of PC after the second dose.

A summary review of carcass cutability data comparing primal value of immunologically and physically castrated barrows.

The objectives were to 1) assess cutability, quality, and value of carcasses from immunologically castrated (IC) barrows compared with carcasses from physically castrated (PC) barrows and 2) evaluate the effect of hot carcass weight (HCW) on cutability and value of IC barrows summarizing U.S. data. Lean cutting yield (LCY) was defined as: LCY = [(whole ham + trimmed loin + Boston butt + picnic + spareribs)/chilled side wt] x 100. Carcass cutting yield (CCY) was determined using the following equation: CCY = [(lean cutting yield components + natural fall belly)/chilled side wt] x 100. To evaluate the effects of HCW of IC barrows on carcass cutting yields, IC barrows were grouped by HCW: light ( < 90.9 kg), average (90.9-97.7 kg), or heavy ( > 97.7 kg). Differences in the value of the carcass components for IC and PC barrow carcasses were calculated using a 5 yr average of meat prices from the USDA Agriculture Marketing Service and the carcass cutting yield estimates generated from this summary. Data were analyzed using the MIXED procedure of SAS with fixed effects of Improvest treatment or HCW group. Study was included as a random effect. This review allowed for a summarization of the treatment averages of 851 IC and PC barrow carcasses. Lean cutting yield of IC barrows was 1.41 units greater (P < 0.0001) than PC barrows (70.97 vs. 69.56%). Similarly, CCY of IC barrows was 1.29 units greater (P < 0.001) compared with PC barrows (87.27 vs. 85.98%). As HCW of IC barrows increased, both CCY and LCY declined (P < 0.01), with light IC barrow carcasses having a 1.43 unit advantage in CCY compared with heavy IC barrow carcasses (P < 0.01). Natural fall bellies of PC barrows comprised a greater (P < 0.05) percentage of side weight than those from IC barrows (15.81 vs. 15.50%). A reduction in belly primal value was confirmed by a 3.43 unit reduction in the commercial bacon slicing yields of IC barrows. However, belly yield and slicing yield differences were minimized when IC barrows were marketed at a heavier weight. Using carcass cutout estimates determined in the summary as the foundation for value calculations, lean cuts of IC barrow carcasses were worth $2.66 to $3.80 more than PC barrow carcasses. Therefore, after adjustment for the reduction in belly primal value, the primal value of an IC barrow carcass was $2.08 to $3.13 greater than a PC barrow carcass.

Ham and belly processing characteristics of immunological castrated barrows (Improvest) fed ractopamine hydrochloride (Paylean).

Effects of sex class (physically castrated, PC or immunologically castrated, IC) and diet (0 or 5mg/kg ractopamine hydrochloride, RAC) on characteristics of ham and bellies were determined from pigs slaughtered in three groups with similar ending live weights. One carcass per pen per marketing group (n=8) was selected to evaluate further processing characteristics. Data were analyzed as a 2×2 factorial design with a split plot in time and fixed effects of sex, diet, marketing group, and their interactions. IC fresh bellies were thinner (P<0.01) and softer (P<0.01) than PC bellies. IC hams and bellies were leaner (P<0.05) than those from PC pigs. RAC feeding did not affect (P>0.05) fresh ham or belly characteristics but decreased (P<0.01) fat in cured PC bellies. Marketing group affected (P<0.05) fresh quality, processing characteristics, and composition of hams and bellies. Immunological castration and RAC produced leaner finished products but did not alter processing yield of hams or bacon.

An apparent connection between histidine, recombination, and repair in Neurospora.

Two mutants of Neurospora crassa, uvs-3 and mei-3, share four properties--UV sensitivity, inhibition by histidine, meiotic blockage when homozygous, and increased duplication instability (due to mitotic crossing over, to deletions or to both). The present paper shows that a third nonallelic mutant, uvs-6, exhibits the same four properties.--Also, the instability of duplications in the absence of any UV-sensitive mutant is increased by the presence of histidine in the growth medium.

Superoxide dismutase (sod-1) null mutants of Neurospora crassa: oxidative stress sensitivity, spontaneous mutation rate and response to mutagens.

Enzymatic superoxide-dismutase activity is believed to be important in defense against the toxic effects of superoxide. Although superoxide dismutases are among the best studied proteins, numerous questions remain concerning the specific biological roles of the various superoxide-dismutase types. In part, this is because the proposed damaging effects of superoxide are manifold, ranging from inactivation of certain metabolic enzymes to DNA damage. Studies with superoxide-deficient mutants have proven valuable, but surprisingly few such studies have been reported. We have constructed and characterized Neurospora crassa mutants that are null for sod-1, the gene that encodes copper-zinc superoxide dismutase. Mutant strains are sensitive to paraquat and elevated oxygen concentrations, and they exhibit an increased spontaneous mutation rate. They appear to have near wild-type sensitive to near- and far-UV, heat shock and gamma-irradiation. Unlike the equivalent Saccharomyces cerevisiae mutant and the sodA sodB double mutant of Escherichia coli, they do not exhibit aerobic auxotrophy. These results are discussed in the context of an attempt to identify consensus phenotypes among superoxide dismutase-deficient mutants. N. crassa sod-1 null mutant strains were also employed in genetic and subcellular fractionation studies. Results support the hypothesis that a single gene (sod-1), located between Fsr-12 and leu-3 on linkage group I, is responsible for most or all CuZn superoxide dismutase activity in this organism.

Color stability and sensory characteristics of fresh and enhanced pork loins from immunologically castrated barrows.

Our objective was to evaluate color changes during storage of fresh pork chops and enhanced and nonenhanced loin quality from anti-gonadotropin-releasing factor (Improvest) immunologically castrated (IC) barrows. In study 1, treatments evaluated were IC barrows, IC barrows fed ractopamine (IC+RAC), physically castrated (PC) barrows, intact males (IM), and gilts. Fresh loins were evaluated for sensory characteristics and instrumental tenderness including both Warner-Bratzler Shear force (WBSF) and star probe force. In study 2, treatments evaluated were IC barrows fed 0.55% and 0.65% standard ileal digestible (SID) lysine and PC barrows fed 0.55% SID lysine. Loin chops were displayed for 7 d, and color was evaluated. The remaining loin was halved, and one half was enhanced with a 3.5% salt and phosphate solution. Enhanced loins were evaluated for sensory characteristics and instrumental tenderness. In both studies, pen served as the experimental unit for all traits measured. Data from individual animals were averaged by pen and analyzed, per study, as a 1-way ANOVA using the MIXED procedure of SAS. In study 1, there were no differences ( 0.05) between treatments for juiciness, tenderness, chewiness, or off-flavor. Intact males had the most intense ( 0.05) boar aroma. Gilts had the most intense ( < 0.05) pork flavor, whereas IM had the least intense pork flavor; all other treatments were intermediate. Tenderness (WBSF) was similar between treatments at 1 d of aging; however, at 7, 14, and 21 d of aging, loins from IC barrows were more tender ( 0.05) than those from gilts, IM, and IC+RAC but were similar ( 0.05) to those from PC. In study 2, discoloration of fresh loin chops during storage was similar ( 0.42) between PC and IC barrows. Chops from enhanced loins were more tender and juicy but had more off-flavor than nonenhanced loins ( 0.01). Star probe and WBSF were also reduced ( 0.01) in enhanced loins compared with nonenhanced loins. Sensory characteristics and tenderness were not different between treatments of IC and PC barrows ( > 0.05), and there were no interactions of enhancement with castration treatments. These data suggest that immunological castration does not negatively impact the color stability, sensory characteristics, or tenderness of enhanced or nonenhanced pork loins.

Evaluation of whole-mount in situ hybridization as a tool for pathway-based toxicological research with early-life stage fathead minnows.

Early-life stage fish can be more sensitive to toxicants than adults, so delineating mechanisms of perturbation of biological pathways by chemicals during this life stage is crucial. Whole-mount in situ hybridization (WISH) paired with quantitative real-time polymerase chain reaction (QPCR) assays can enhance pathway-based analyses through determination of specific tissues where changes in gene expression are occurring. While WISH has frequently been used in zebrafish (Danio rerio), this technology has not previously been applied to fathead minnows (Pimephales promelas), another well-established small fish model species. The objective of the present study was to adapt WISH to fathead minnow embryos and larvae, and use the approach to evaluate the effects of estrone, an environmentally-relevant estrogen receptor (ER) agonist. Embryos were exposed via the water to 0, 18 or 1800 ng estrone/L (0, 0.067 and 6.7nM) for 3 or 6 days in a solvent-free, flow-through test system. Relative transcript abundance of three estrogen-responsive genes, estrogen receptor-α (esr1), cytochrome P450-aromatase B (cyp19b), and vitellogenin (vtg) was examined in pooled whole embryos using QPCR, and the spatial distribution of up-regulated gene transcripts was examined in individual fish using WISH. After 3 days of exposure to 1800 ng estrone/L, esr1 and cyp19b were significantly up-regulated, while vtg mRNA expression was not affected. After 6 days of exposure to 1800 ng estrone/L, transcripts for all three genes were significantly up-regulated. Corresponding WISH assays revealed spatial distribution of esr1 and vtg in the liver region, an observation consistent with activation of the hepatic ER. This study clearly demonstrates the potential utility of WISH, in conjunction with QPCR, to examine the mechanistic basis of the effects of toxicants on early-life stage fathead minnows.

The effects of ionizing radiation on DNA synthesis in eukaryotic cells.

PURPOSE: To review observations of the effects of ionizing radiation on DNA synthesis in eukaryotes. CONTENT: Available information broadly falls into two categories: descriptions of the phenomenon, including dose response data and analysis; and, more recently, investigations utilizing genetic approaches. The down-regulation of DNA replication in the presence of radiation-induced DNA damage appears to be an active cellular response, termed the S-phase damage-sensing (SDS) checkpoint control (Larner et al. 1997). Observations on a variety of eukaryotes, including man, suggest that the regulatory controls involved are highly conserved and may additionally function in G1 and G2 checkpoint controls. Budding yeast, fission yeast and human homologues are identified. CONCLUSIONS: The SDS checkpoint control appears to be comprised of a complex of checkpoint proteins that respond to the stalled replication complex. The replication complex is thought to signal down-regulation of the mitotic kinase, ensuring that the cell does not enter mitosis while S phase is delayed. Concomitantly, the checkpoint complex is believed to transmit a signal via two key checkpoint proteins (Rad3 and Cds1 in the fission yeast), in order to arrest further DNA synthesis initiation.

Damage-resistant DNA synthesis in eukaryotes.

The molecular basis of sensitivity of ionizing radiation and other damaging agents is not clearly defined in eukaryotes. While a large number of mutants have been described only a few have been demonstrated to have a defect in the repair of damage to DNA. An interesting characteristic of a sub-group of these mutants, in different species extending throughout the phylogenetic scale, is the presence of damage-resistant DNA synthesis. This phenomenon is observed in cells from individuals with the genetic disorder ataxia telangiectasia, in HeLa cells treated with fluorodeoxyuridine prior to UV irradiation, in mutants of the fungus Neurospora crassa, the slime mould Dictyostelium discoideum, the fruit fly Drosophila melanogaster and possibly in the "wasted" mouse mutant. In the case of ataxia telangiectasia sensitivity is only observed to ionizing radiation or radiomimetic chemicals whereas sensitivity to a wider spectrum of mutagens is reported for the lower eukaryotic mutants. In all cases a reduced inhibition of DNA synthesis is obtained after exposure to an agent to which the cell type is hypersensitive. It is unclear how damage-resistant DNA synthesis contributes to increased sensitivity in these cells, but is unlikely to be the major mechanism predisposing to radiation-induced cell death. The description of a derivative of an ataxia telangiectasia cell line with normal sensitivity to radiation but still maintaining resistant DNA synthesis partially uncouples radioresistant DNA synthesis and radiosensitivity. This paper is designed to review the phenomenon of damage-resistant DNA synthesis in a number of mutants.

A new mutagen-sensitive mutant in Neurospora, mus-16.

A new gene, mus-16, is determined by the nitrogen mustard-sensitive Neurospora mutant of Baker, Parish and Curtis (1984) which is defective in the removal of DNA-DNA and DNA-protein crosslinks. This gene is on the left arm of linkage group V between caf-1 and lys-1. The mus-16(JMB) mutant is sensitive to the alkylating agents methyl methanesulfonate (MMS) [dose reduction factor (drf) 8-10 X], N-methyl-N'-nitro-N-nitrosoguanidine (drf 5-6 X), the amino acid histidine and the drug hydroxyurea. It is not sensitive to ultraviolet-light, gamma-irradiation, or mitomycin C (MMC). It shows normal spontaneous mutation rates but increased induction of mutation by MMS. Homozygous crosses are barren, showing no signs of sporulation. Mitotic spontaneous chromosome instability is increased. The mus-16 mutation is similar to several non-excision repair-defective mutants in Neurospora. Some of these may be defective in repair of alkylation damage. The MMC data supports earlier data that in fungi MMC is incapable of forming DNA-DNA crosslinks.

Gamma-ray-sensitive mutants of Neurospora crassa with characteristics analogous to ataxia telangiectasia cell lines.

Well characterized gamma-ray sensitive mutants of the fungus Neurospora crassa have been screened for characteristics analogous to those of cell lines derived from humans with the genetic disease, ataxia telangiectasia (AT). Two Neurospora mutants, uvs-6 and mus-9, show the AT cell line characteristics of gamma-ray and bleomycin sensitivity, and little or no repression of DNA synthesis following treatment with these agents. Normal human or Neurospora cells show an extensive biphasic DNA synthesis repression (to 50% of control) and when DNA synthesis is analyzed by alkaline sucrose gradient centrifugation, repression of DNA synthesis by low doses of gamma-radiation occurs primarily in low molecular weight (MW) DNA pieces in both organisms. In AT cells and the uvs-6 mutant, no repression in synthesis of low or higher MW DNA is seen at low doses, while the mus-9 mutant shows little repression of high MW DNA, but an intermediate level of low MW DNA synthesis. Both mutants have been shown previously to have an increased level of spontaneous chromosome instability as do AT lines. The uvs-6 and mus-9 mutations are known to be due to two different genes in two different epistatic groups. These results demonstrate that AT-like cellular characteristics can arise from defects in at least two and probably any of several genes, and that lower eukaryotes such as Neurospora can provide an inexpensive and useful model for AT while avoiding the problems inherent in using transformed cell lines.

Deoxyribonucleoside triphosphate pools in Neurospora crassa: effects of histidine and hydroxyurea.

An effective HPLC method for detecting deoxyribonucleoside triphosphates in hyphae from the fungus Neurospora crassa has been developed. In rapidly growing cells the nucleotide levels vary from 11.8 pmoles/micrograms DNA for dGTP to 24.2 pmoles/micrograms DNA for dTTP. These levels fall by approximately one half in stationary-phase cultures but the ratio of each pool to dGTP remains the same. The dNTP pools in conidia are at least 5-fold lower than in rapidly growing cells. The pool sizes are the same in static and shaking cultures. When the ribonucleotide reductase inhibitor, hydroxyurea (30 mM), is added to rapidly growing cultures, DNA synthesis is stopped and the dGTP pool is reduced by 39%, while the size of the other pools remains the same. In the presence of 11 mM histidine, DNA synthesis is also stopped and the size of the dGTP pool reduced by 46% while the deoxypyrimidine pools are somewhat increased. This suggests that the toxicity of excess histidine in Neurospora may be due to its ability to interact with the ribonucleotide reductase, inactivating the enzyme. Histidine may react with the free radical at the active site, as does hydroxyurea.

Growth, carcass composition and plasma melatonin in postpubertal beef heifers fed melatonin.

Two experiments were conducted to determine if feeding melatonin alters plasma concentrations of melatonin, growth and carcass composition of postpubertal beef heifers exposed to 16 h light (L):8 h dark (D). In Exp. 1, 16 heifers were blocked by initial body weight (318 +/- 5.6 kg). Four heifers were killed before starting the melatonin treatment to obtain initial carcass composition. Six heifers received vehicle (95% ethanol) and six were fed melatonin (4 mg/100 kg body weight) daily for 58 d at 1330 to coincide with the middle of the 16-h light period. On d 59 heifers were slaughtered. Melatonin feeding increased the percentage of fat in rib (P less than .05) and longissimus muscle (LD; P less than .10) and carcass fat accretion 28% (P less than .09) but reduced the percentage of protein 8% in rib (P less than .05) and carcass protein accretion 30% (P less than .09). Other measures in the carcass and body weight gain were not affected (P greater than .10) by feeding melatonin. Plasma concentrations of melatonin increased (P less than .01) from 10 to 140 pg/ml within 30 min of feeding melatonin. In Exp. 2, 24 heifers were blocked by initial body weight (348 +/- 13.7 kg). Eight heifers were killed initially, eight received vehicle and eight were fed melatonin for 63 d as described in Exp. 1. Melatonin did not influence (P greater than .10) body weight gain or any measure in the carcass; however, these heifers were fatter (40.1%) than those in Exp. 1 (30.9%) at the beginning of the experiment.(ABSTRACT TRUNCATED AT 250 WORDS)

Measurement of protein turnover in skeletal muscle strips.

Isolated porcine and bovine muscle strips were incubated in Krebs Ringer bicarbonate buffer to determine in vitro protein synthesis (PS) and protein degradation (PD) rates to validate the in vitro system for use with livestock species. The addition of 5X plasma concentrations of amino acids to the medium stimulated PS 30%. Addition of 3.5 mM leucine to a leucine-deficient buffer supplemented with amino acids decreased PD 37% and stimulated PS 24%. The addition of .1 U/ml insulin reduced PD 28% and increased PS 30%. Protein degradation was elevated in longitudinally split rat soleus and extensor digitorum longus muscles compared to their contralateral intact muscles. Muscle strips must be removed within 15 min of exsanguination because PD rates become greatly elevated thereafter. ATP concentrations declined during incubation, but the addition of ATP or creatine had no effect on either PD or PS. Neither PD nor PS was affected by the addition of transferrin, fetuin, ascorbate, dexamethasone or indomethacin to the incubation medium. However, muscle strips were sensitive to the addition of triiodothyronine (T3), PD was increased up to 75% as T3 concentration was increased, and PS rates doubled compared to controls. Serum from mature barrows or gilts had no effect on protein turnover, but the addition of 10% and 15% serum from boars increased both PD and PS. With fasted pigs a continual decline in PS occurred over 5 d, whereas PD was elevated at 3 d and then declined to rates comparable to the fed state after 5 d. These data suggest that the in vitro system has application for assessing relative changes that occur in vivo following nutritional, physiological and endocrinological manipulation.

Effects of immunological castration (Improvest) on further processed belly characteristics and commercial bacon slicing yields of finishing pigs.

Objectives were to compare fresh belly characteristics, further processed belly characteristics, and commercial bacon slicing yields of immunologically castrated (IC) barrows, IC barrows fed ractopamine hydrochloride (IC+RAC), physically castrated (PC) barrows, intact males, and gilts. One hundred eighty-eight bellies from pigs housed in single sex pens (n = 48) slaughtered at 130 kg ending live weight were evaluated for flop distance, length, width, thickness, and fatty acid composition. Bellies were injected, thermally processed, and sliced according to standard protocols at a USDA federally inspected facility. Complete slices were sorted by trained plant personnel. Then, sliced bellies were individually packaged to maintain anatomical orientation. The effects of treatments were analyzed as a generalized linear mixed model with pen of pigs serving as the experimental unit for all comparisons. Belly thickness was not different (P ≥ 0.11) in bellies from IC barrows (3.74 cm) compared with bellies from IC+RAC (3.60 cm), PC barrows (3.94 cm), or gilts (3.64 cm); however, bellies were 0.42 cm thicker (P < 0.01) in IC barrows compared with intact males. Iodine value was not different (P ≥ 0.06) in bellies from IC barrows (68.26) compared with bellies from IC+RAC (69.78), PC barrows (67.55), or gilts (69.45); however, iodine value was 2.32 units less (P < 0.01) in IC barrows compared with intact males. Green weight was not different (P ≥ 0.09) in bellies from IC barrows (4.88 kg) compared with bellies from IC+RAC (5.11 kg), PC barrows (5.10 kg), or gilts (4.79 kg); however, green weight was 0.56 kg greater (P < 0.0001) in IC barrows compared with intact males. Pump uptake and cooked yield was not different (P ≥ 0.15) among treatment groups. Slicing yield (as a percentage of green weight) was decreased (P ≤ 0.01) in bellies from IC barrows (93.61%) compared with bellies from PC barrows (98.42%) or gilts (98.19%); however, slicing yield was not different (P ≥ 0.11) among bellies from IC barrows, IC+RAC, or intact males. Overall, fresh belly characteristics of IC barrows were analogous to PC barrows and gilts, but commercial slicing yields of bacon manufactured from IC barrows were less than both PC barrows and gilts.

Effects of immunocastration and time after second Improvest dose on adipose tissue fatty acid profile of finishing barrows.

The effects of immunocastration and time after second Improvest dose on proximate composition and fatty acid profile of adipose tissue from jowl and belly of finishing barrows were determined. Physically castrated (PC) and immunologically castrated (IC) barrows were assigned to treatments at birth. Within 5 d of age, PC barrows were physically castrated and IC barrows were administered Improvest (gonadotropin-releasing factor analog; Zoetis, Kalamazoo, MI) at 16 and 20 wk of age. Diets were formulated with corn and soybean meal and did not contain ethanol coproducts. Subsequently, PC (n = 23) and IC (n = 24) barrows were slaughtered biweekly from 22 to 28 wk of age, 2 to 8 wk following second Improvest dose. Adipose tissue samples were collected from the jowl and belly. Main effects of castration method and time after second Improvest dose and their interaction were analyzed with the MIXED procedure of SAS. Overall, IC barrows were heavier at slaughter (136.1 vs. 100.6 kg; P < 0.01), had similar HCW (P = 0.13), and were leaner (50.9 vs. 48.9%; P < 0.01) than PC barrows. In jowl adipose tissue, IC barrows had less lipid content (64.7 vs. 80.2%; P < 0.01) than PC barrows at 2 wk. However, there were no differences (P ≥ 0.75) in lipid content between IC and PC barrows from 4 to 8 wk. Total MUFA content was reduced (42.5 vs. 44.8%; P = 0.04) and PUFA content was greater (16.9 vs. 14.8%; P < 0.01) in IC compared to PC barrows. Increasing time after second dose reduced (P = 0.04) PUFA content from 17.5% at 2 wk to 15.4% at 8 wk. In IC barrows, iodine value (IV) was increased by 6.9 and 3.3 g/100 g compared with PC barrows at 2 and 4 wk, but IV was not different between castration methods (P ≥ 0.95) at both 6 and 8 wk. Meanwhile, in belly adipose tissue, IC barrows had less lipid content than PC barrows (75.3 vs. 82.7%; P < 0.01). Lipid content was unchanged over time (P = 0.34) in either PC or IC barrows. Total SFA and MUFA content of the belly were similar for both IC and PC barrows. Belly adipose tissue from IC barrows had greater concentrations of PUFA than PC barrows (16.2 vs. 14.7%; P < 0.01). In IC barrows, IV was increased by 5.3 g/100 g compared with PC barrows at 2 wk, but IV was not different between castration methods (P ≥ 0.91) from 4 to 8 wk. Results indicate that as time after second Improvest dose progressed, IV of jowl and belly was reduced. This was accompanied by an increase in lipid deposition in jowl, while lipid content remained constant in belly.

Effects of ractopamine on growth performance and carcass characteristics of immunologically and physically castrated barrows and gilts.

The study was performed to evaluate the effect of feeding ractopamine (RAC) to physically castrated barrows (PC), immunologically castrated barrows (IC), and gilts (gilts) using a randomized complete block design with a 3 × 3 factorial arrangement of treatments: 1) sex (PC, IC, and gilts) and 2) RAC inclusion level (0, 5, and 7.5 mg/kg). The IC received 2 doses of gonadotropin releasing factor analog-diphtheria toxoid conjugate (Improvest; Zoetis, Kalamazoo, MI) at the start of the study (wk 16 of age; 69.6 ± 2.96 kg BW) and 4 wk later. The study used 180 pigs housed in groups of 4 (5 groups/sex × RAC subclass) and was performed over a fixed time of 61 d with RAC being fed for the final 26 d of study. Diets were formulated to meet requirements of intact males for the first 35 d and of intact males fed 7.5 mg/kg RAC for the remainder of the study. Pigs had ad libitum access to feed and water throughout the study period. At the end of the study, pigs were harvested at a commercial facility and HCW and last rib backfat thickness were measured. There were no treatment interactions (P > 0.05) for any variables. For the overall study period, IC had greater (P ≤ 0.05) overall ADG compared to PC, which grew faster (P ≤ 0.05) than gilts (1,246, 1,083, and 1,025 g/d for IC, PC, and gilts, respectively; SEM = 20.3); ADFI was lower (P ≤ 0.05) for gilts than IC and PC, which had similar ADFI (3.36, 3.37, and 2.87 kg/d, respectively; SEM = 0.051); and G:F was greater (P ≤ 0.05) for IC than gilts and greater for gilts than PC (0.371, 0.322, and 0.358 kg/kg, respectively; SEM = 0.0039). For the period from the second dose to the end of study, IC had greater (P ≤ 0.05) ADG (28.6%), ADFI (12.3%), and G:F (14.3%) than PC. Carcass yield was lower (P ≤ 0.05) for IC compared to PC and gilts (72.8, 75.0, and 74.6%, respectively; SEM = 0.25). Feeding RAC increased (P ≤ 0.05) ADG (15.7 and 14.5% for 5 and 7.5 mg/kg, respectively), G:F (17.1 and 16.4%, respectively), carcass weight (3.7 and 3.2 kg, respectively), and carcass yield (1.0 and 1.0 percentage unit, respectively) compared to the control. These results highlight sex differences in and effects of RAC on growth and carcass characteristics and suggest that the relative response to RAC is similar in IC and PC.

Growth performance of immunologically castrated (with Improvest) barrows (with or without ractopamine) compared to gilt, physically castrated barrow, and intact male pigs.

The study used a randomized complete block design (blocking factor was date of start on test) with 5 treatments: 1) physically castrated barrows (PC), 2) intact males (IM), 3) gilts (G), 4) immunologically castrated barrows (IC), and 5) immunologically castrated barrows fed ractopamine at 5 mg/kg (IC+RAC). The study used 192 pigs and was performed from the 16 wk of age (67.2 ± 2.52 kg BW) to a pen mean BW of 132.5 ± 3.60 kg. For IC+RAC, ractopamine was fed for the final 23 d of the study. Pigs were housed in groups of 4 (10 groups for PC, IM, G, and IC and 8 groups for IC+RAC) in a finishing building at a floor space of 1.18 m(2)/pig. Diets were formulated to meet requirements of IM except that the diet for the IC+RAC fed during the ractopamine feeding period was formulated to meet requirements of pigs on that treatment. Pigs had ad libitum access to feed and water throughout the study period and were individually weighed at the start, wk 2 and 4, and subsequently every week until the end of study. For the overall study period, IC had greater (P ≤ 0.05) ADG than the other genders (1,150, 1,024, 1,064, and 954 g/d for IC, PC, IM, and G, respectively; SEM = 25.8) and required fewer days to reach slaughter weight than the other genders (58.1, 61.6, 61.6, and 66.5 d for IC, PC, IM, and G, respectively; SEM = 1.26). Overall ADFI was less (P ≤ 0.05) for IM and G than IC and PC, which were similar (P > 0.05) in this respect (3.11, 3.06, 2.68, and 2.75 kg/d for IC, PC, IM, and G, respectively; SEM = 0.061). Overall G:F was greater (P ≤ 0.05) for IM than the other genders; IC had greater overall G:F than PC and G, which were similar in this respect (0.371, 0.335, 0.397, and 0.347 kg/kg for IC, PC, IM, and G, respectively; SEM = 0.0068). Immunologically castrated barrows had greater (P ≤ 0.05) ADG (30.7%) and ADFI (22.5%) than PC from the second week following the second Improvest dose to the end of the study. During the ractopamine feeding period, IC+RAC had greater (P ≤ 0.05) ADG (11.6%) and G:F (17.3%) than IC. The results of this study confirmed previously observed gender differences and effects of ractopamine on growth performance and that IC grew faster and had greater feed efficiency than PC during the study period.