Storage length, storage temperature, and lean formulation influence the shelf-life and stability of traditionally packaged ground beef.

The effect of storage length and temperature on the shelf life of three ground beef formulations (lean:fat: 73:27, 81:19 and 91:9) was investigated. Coarsely ground beef was stored at -1.7 or 2.3°C for up to 28d. Traditional overwrap packages were produced every 7d prior to retail display for 24h. Lipid oxidation (TBARS), subjective color, instrumental color, and aerobic bacteria were evaluated after 0 and 24h of display. Formulation influenced initial L* and subjective color values (P<0.05). Storage temperature did not affect initial color, but product stored at 2.3°C was more discolored after 24h (P<0.05). Aerobic bacteria increased as storage d and temperature increased (P<0.05). Initial TBARS increased through d 21, but were lower after 28d. Overall, initial characteristics depended on formulation; however, ground beef shelf-life and stability were largely influenced by storage length and storage temperature.

Effect of castration and dehorning singularly or combined on the behavior and physiology of Holstein calves.

The objectives of this study were to determine i) the effect of castration, dehorning, or both on the physiology and behavior of 3-mo-old Holstein calves, and ii) the effectiveness of pain relief to alleviate the pain caused by castration and/or dehorning. Holstein calves (n = 80) were assigned randomly to 1 of 8 treatments (10 calves/treatment): i) control handling (SHAM); ii) surgical castration (CAS); iii) dehorning (DH); iv) surgical castration and dehorning (CD); v) control handling plus pain relief (ANA); vi) surgical castration plus pain relief (CAS+A); vii) dehorning plus pain relief (DH+A); or viii) surgical castration and dehorning plus pain relief (CD+A). Pain relief consisted of administering local anesthetic and a nonsteroidal anti-inflammatory drug (NSAID) immediately before castration, dehorning, or both. Sequential blood samples were collected to measure leukocyte counts and cortisol concentrations. Behavior was recorded using 5-min scan samples during the first 3 h after application of the treatments. Calves were weighed before and 24 h after treatment application. Calves dehorned spent more time head shaking (p < 0.001) and ear flicking (p < 0.05), and CD calves spent more time ear flicking (p < 0.05) and foot stamping (p < 0.01) than SHAM handled calves. Calves castrated, dehorned, or both spent less (p < 0.01) time eating compared with sham handled calves. Giving calves pain relief before castration and/or dehorning increased (p < 0.05) the time spent eating compared with CAS, DH, and CD calves. At 6 h posttreatment, neutrophil to lymphocyte ratio was greater (p < 0.01) in castrated and/or dehorned calves compared with SHAM-handled calves. Castration and/or dehorning also increased (p < 0.05) cortisol concentrations for at least 4 h after these procedures were performed; however, administering pain relief before castration and/or dehorning markedly reduced (p < 0.05) this response. Behavioral and physiological changes caused by castration, dehorning, or both are indicative of calves experiencing pain for at least 4 h after application of these procedures, and these responses were additive when performed together. Therefore, providing calves with pain relief, in the form of local anesthetic and an NSAID, can markedly reduce both the behavioral and physiological response to these procedures.

Administration of anesthetic and analgesic prevent the suppression of many leukocyte responses following surgical castration and physical dehorning.

The objectives of the current research were to determine the physiological effects and responses of many leukocytes following surgical castration and/(or) physical dehorning and the influence of anesthetics and analgesics in 3-month-old calves. Eighty 3-month-old Holstein bull calves were completely randomized to treatments in a 2 × 2 × 2 factorial arrangement with castration, dehorning, and anesthetic/analgesic as the main effects. Peripheral blood samples were collected just before (0) and 0.5, 1.5, 2.5, 4, 6, 24, and 72 h after the respective procedure(s) and analyzed for total leukocyte and differential counts, as well as plasma cortisol and haptoglobin concentrations. Blood from the 0, 0.5 and 24h collections were analyzed for many ex vivo leukocyte responses. Data were analyzed using a repeated measures analysis of variance with the fixed effects of treatment, time, and the interaction of treatment × time. Pre-planned contrasts were performed to determine the effect of (1) management procedure (castration and/(or) dehorning), (2) anesthetic/analgesic, and (3) were the management procedures additive. There were treatment × time interactions (P<0.05) on plasma cortisol and haptoglobin concentrations as well as for total leukocyte and neutrophil concentrations in blood. Castration and dehorning increased cortisol concentrations and the effect of the procedures was additive (P<0.02). Dehorning alone elicited a greater (P<0.05) cortisol response than castration alone. In contrast, the leukocytosis and neutrophilia was greater (P<0.01) among castrated calves. In addition, haptoglobin concentrations at 24h after castration were elevated (P<0.01) in calves that were castrated. Both castration and dehorning suppressed (P=0.04) many leukocyte responses including the secretion of tumor necrosis factor-α when whole blood cultures were stimulated with lipopolysaccharide, surface expression of L-selectin on peripheral blood neutrophils, and the oxidative burst intensity of peripheral blood neutrophils when co-cultured with an Escherichia coli. The effects of castration and dehorning on blood leukocyte counts or any of the leukocyte responses were not additive (P>0.23). Castration and dehorning effects of plasma haptoglobin concentrations tended (P=0.10) to be additive at 72 h after the procedure(s). Prior administration of local anesthetic and a systemic analgesic attenuated (P<0.001) the cortisol response and prevented (P=0.03) the observed leukocytosis, neutrophilia, and leukocyte suppression. These data suggest that calves should be castrated and dehorned on the same day rather than spreading them out across two days and calves should be administered pain relief prior to performing either procedure.

The physiological and behavioral response of pigs castrated with and without anesthesia or analgesia.

Surgical castration is a common management practice performed on male pigs to prevent the occurrence of boar taint. Surgical castration is known to cause physiological and behavioral changes in pigs indicative of pain-induced distress; however, it is commonly performed without pain relief. The objective of this study was to evaluate the effectiveness of carbon dioxide gas (CO(2)) anesthesia and a non-steroidal anti-inflammatory drug (NSAID) to alleviate the pain caused by castration. At 3 d of age, male pigs were either control handled (CON), castrated without pain relief (CAS), given an NSAID and then immediately castrated (CAS+NSAID), anesthetized with CO(2) and then castrated (CAS+CO2), or anesthetized with CO(2) and given an NSAID at the time of castration (CAS+BOTH). Blood samples were collected before castration, and at 30, 60, 120, and 180 min, 24 h, and 3 d after castration or handling for analysis of cortisol, C-Reactive protein (CRP), and substance-P (SP) concentrations. This study was then repeated using the same treatment groups, and the behavioral response to castration and handling were measured using a 1-min scan sampling procedure. The percentage of stress vocalizations was recorded during the administration of all treatments. Anesthesia and analgesia did not effectively reduce (P > 0.05) the cortisol response to surgical castration. Overall, CRP concentrations were greater (P < 0.05) in CAS+CO2 pigs as compared with CON pigs. Sixty minutes after castration or handling, SP concentrations were greater (P < 0.08) in pigs given CO(2) anesthesia (CO2, CAS+CO2, and CAS+BOTH) than CON, CAS, and CAS+NSAID pigs. Pigs castrated without pain relief spent more (P < 0.001) time lying without contact than all other treatments during the first 30 min after castration, but thereafter CAS+CO2 pigs spent more (P < 0.001) time lying without contact than other treatments. During the first 30 min after the treatments were applied, CAS+CO2 pigs spent more (P < 0.01) time displaying pain-like behaviors than CON, CAS, CAS+NSAID, and CAS+BOTH pigs. The percentage of stress vocalizations was greater (P < 0.05) in CAS and CAS+NSAID pigs than all other treatments. Neither CO(2) anesthesia nor a NSAID, given separately or combined, markedly reduced the pain-induced distress caused by castration in pigs. More research is needed to evaluate practical methods of on-farm pain relief for pigs.

Physiology and behavior of pigs before and after castration: effects of two topical anesthetics.

Surgical castration of male piglets is a common management practice conducted on commercial swine farms to prevent the occurrence of boar taint and aggressive behavior. However, the procedure of surgical castration causes acute pain-induced distress, which is an animal welfare concern. The objective of this study was to evaluate the use of two topical anesthetics to alleviate the pain caused by castration in piglets as measured by physiological and behavior indices of stress. At 3 days of age, 40 weight-matched piglets were allocated to one of four treatment groups. Treatments included: (i) sham castration (CON), (ii) surgical castration (CAS), (iii) castration and short-acting local anesthetic applied topically to the castration wound (SHORT) and (iv) castration and long-acting local anesthetic applied topically to the castration wound (LONG). Blood samples were collected from piglets before and 30, 60, 120 and 180 min after castration to measure leukocyte and differential counts and cortisol concentrations. The above experiment was repeated without blood collection and behavior was recorded for 30 min before and 180 min after castration or handling. Stress vocalizations were recorded during castration and handling. All piglets were weighed before and 24 h after castration and wound healing was recorded daily for the first 14 days after castration. Leukocyte counts and differentials did not differ (P > 0.05) among any of the treatments. Cortisol concentrations were elevated (P < 0.06) in CAS, SHORT and LONG piglets compared with controls 30 and 60 min after castration. The percentage of stress vocalizations was greater (P < 0.05) among castrated piglets compared with CON piglets, regardless of anesthetic treatment. Piglets that were castrated and not given a topical anesthetic spent more time (P < 0.05) lying without contact compared with piglets castrated and given a topical anesthetic, regardless of the topical anesthetic administered. Body weight change did not differ (P > 0.05) among treatments 24 h after castration or control handling and wound healing scores were greater (P < 0.05) in SHORT compared with CAS and LONG piglets 9 to 14 days after castration. In this study, the use of a short- or long-acting topical anesthetic was not effective in reducing the pain-induced distress caused by castration in piglets. Further research is needed to evaluate alternative practical methods to reduce the pain caused by the on-farm castration of piglets.

Chronic inflammatory pain and the neurovascular unit: a central role for glia in maintaining BBB integrity?

Pain is a complex phenomenon involving both a peripheral innate immune response and a CNS response as well as activation of the hypothalamic-pituitary-adrenal axis. The peripheral innate immune response to injury involves the rapid production and local release of proinflammatory cytokines such as tumor necrosis factor-alpha (TNF-/alpha), interleukin-1 (IL-1) and IL-6. Recent studies into the CNS response to peripheral chronic inflammatory pain strongly implicates a role for glia, and local synthesis of proinflammatory cytokines and growth factors. A characteristic feature of CNS inflammation is gliosis, in which inflammatory mediators activate glial cells (e.g. astrocytes and microglia, macrophages and leukocytes) which have been shown to induce and maintain hyperalgesia. In addition, inflammatory pain induces changes in blood-brain barrier (BBB) permeability and alters transport of clinically relevant drugs used to treat pain into the brain. Despite the increasing body of evidence for the involvement of glia in chronic pain and the role of glia in maintaining the BBB, few studies have addressed glial/endothelial interactions and the mechanisms by which glia may regulate the BBB during inflammatory pain. Further studies into the cellular mechanisms of glial/endothelial interactions may identify novel therapeutic targets for reversing chronic inflammatory induced BBB dysfunction and innovate therapies for modulating the severity of chronic inflammatory pain.

Construction and characterization of an effector strain of Streptococcus mutans for replacement therapy of dental caries.

An effector strain has been constructed for use in the replacement therapy of dental caries. Recombinant DNA methods were used to make the Streptococcus mutans supercolonizing strain, JH1140, lactate dehydrogenase deficient by deleting virtually all of the ldh open reading frame (ORF). To compensate for the resulting metabolic imbalance, a supplemental alcohol dehydrogenase activity was introduced by substituting the adhB ORF from Zymomonas mobilis in place of the deleted ldh ORF. The resulting clone, BCS3-L1, was found to produce no detectable lactic acid during growth on a variety of carbon sources, and it produced significantly less total acid due to its increased production of ethanol and acetoin. BCS3-L1 was significantly less cariogenic than JH1140 in both gnotobiotic- and conventional-rodent models. It colonized the teeth of conventional rats as well as JH1140 in both aggressive-displacement and preemptive-colonization models. No gross or microscopic abnormalities of major organs were associated with oral colonization of rats with BCS3-L1 for 6 months. Acid-producing revertants of BCS3-L1 were not observed in samples taken from infected animals (reversion frequency, <10(-3)) or by screening cultures grown in vitro, where no revertants were observed among 10(5) colonies examined on pH indicator medium. The reduced pathogenic potential of BCS3-L1, its strong colonization potential, and its genetic stability suggest that this strain is well suited to serve as an effector strain in the replacement therapy of dental caries in humans.

Genetic and biochemical analysis of mutacin 1140, a lantibiotic from Streptococcus mutans.

Streptococcus mutans JH1000 and its derivatives were previously shown (J. D. Hillman, K. P. Johnson, and B. I. Yaphe, Infect. Immun. 44:141-144, 1984) to produce a low-molecular-weight, broad-spectrum bacteriocin-like inhibitory substance (BLIS). The thermosensitive vector pTV1-OK harboring Tn917 was used to isolate a BLIS-deficient mutant, DM25, and the mutated gene was recovered by shotgun cloning in Escherichia coli. Sequence analysis of insert DNA adjacent to Tn917 led to the identification of four open reading frames including two (lanA and lanB) which have substantial homology to the Staphylococcus epidermidis structural gene (epiA) and a modifying enzyme gene (epiB) for biosynthesis of the lantibiotic epidermin, respectively. Although the BLIS activity could not be recovered from broth cultures, high yields were obtained from a solid medium consisting of Todd-Hewitt broth containing 0.5% agarose that was stab inoculated with JH1140 (a spontaneous mutant of JH1000 that produces threefold-elevated amounts of activity). Agar could not substitute for agarose. Chloroform extraction of the spent medium produced a fraction which yielded two major bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The faster-migrating band was absent in chloroform extracts of the mutant, DM25. The amino acid sequence of this band was determined by Edman sequencing and mass spectroscopy. The results showed that it is a lantibiotic, which we have named mutacin 1140, and that the sequence corresponded to that deduced from the lanA sequence. We observed a number of similarities of mutacin 1140 to epidermin and an S. mutans lantibiotic, B-Ny266, but it appears to have significant differences in the positions of its thioether bridges. It also has other unique features with regard to its leader sequence and posttranslational modification. A proposed structure for mutacin 1140 is presented.

Legal and regulatory issues in hospice care.

The philosophy of hospice care is to improve the quality of the patient's life by alleviating pain; controlling symptoms; and caring for the emotional, physical, and social needs of the dying patient. This concept presents new medicolegal problems with regard to regulation and reimbursement.