An Experimental Infection Model in Sheep and Goats to Evaluate Salmonella Colonization in Deep Tissue Lymph Nodes and after Carcass Vascular Rinsing with Bacteriophages in Goats.

An animal infection model was evaluated on sheep and goats to confirm which species infected with Salmonella enterica serovar Enteritidis C StR (SE13) would provide a consistent and high frequency of Salmonella colonization in lymph nodes (LNs) without causing undue animal morbidity. Sheep and goats (n = 5) were intradermally inoculated with Salmonella, postincubated for 7 days, and euthanized. Superficial cervical, medial iliac, subiliac, mammary, and popliteal LNs were excised from each carcass. Goat LNs had approximately 53% greater Salmonella level compared to sheep. Also, Salmonella was inconsistently recovered from the sheep LNs. Thus, goats were selected to determine the ability of carcass vascular rinsing (with and without bacteriophages) to reduce Salmonella in infected LNs. Goats with similar characteristics were grouped together before being randomly assigned to 3 postharvest treatments; control (CN, not vascularly rinsed; n = 10), vascularly rinsed with a standard Rinse & Chill® solution (RC; 98.5% water and a blend of saccharides and phosphates; n = 10), or vascularly rinsed with a standard Rinse & Chill® solution plus the addition of bacteriophages (BP; n = 10). Rinse & Chill® system was able to successfully deliver a mean 7.0 log PFU/g to the S. Enteritidis-infected LNs (mean 3.5 log CFU/g). However, neither Rinse & Chill® without bacteriophages nor with bacteriophages caused Salmonella reduction (P > 0.05) compared to the nonrinsed goat carcasses.

Use of a Calving Blind That Imitates a Natural Environment.

Many indoor-housed cows isolate at calving when given the opportunity, and calving behaviors vary by blind and pen design. The objectives of this study were to determine if cows preferred calving in a visibly separated (blind) or an open area of a group maternity pen, and if there was a preference for the degree of seclusion provided by the blind (50% vs. 100% coverage). Two calving blinds were provided in a group calving pen, and the amount of visibility through the blinds was created using firehoses secured from the top of a metal frame that lined the entire front of the blind (100%) or with every other hose rolled up (50%). Holstein cows and heifers (n = 79) were enrolled into a dynamic group calving pen 21 ± 3 d before calving. Calving location, the difference in blind use prior to calving compared to a baseline period, and social behaviors were recorded using video observation. There was no difference in the number of cows that calved in or outside of a blind (28 vs. 37 calvings, respectively). Cows were more likely to calve in a blind during the day than at night and as the number of cows in the pen increased. For cows who calved in a blind, there was no preference for calving in the 50% or 100% blind (10 vs. 18, respectively). Providing a varied environment for intensively managed cattle can improve their welfare by allowing cows the opportunity to perform natural behaviors and choice over their environment.

Quantification of cooling effects on basic tissue measurements and exposed cross-sectional brain area of cadaver heads from Holstein cows > 30 mo of age.

Penetrating captive bolt (PCB) is the primary method of preslaughter stunning for cattle and is also used for on-farm euthanasia. The objective of this study was to quantify the impact of cooling on the soft tissue thickness, cranial thickness, total tissue thickness, and cross-sectional brain area of cadaver heads collected from mature (> 30 mo of age) dairy cows following the application of a PCB stun in a frontal placement. Hide-on cadaver heads were obtained from culled dairy cows (N = 37) stunned in a frontal location using a handheld PCB device (Jarvis Model PAS-Type C 0.25R Caliber Captive Bolt, Long Bolt) at a commercial slaughter establishment. Following transport to the University of Wisconsin-River Falls, heads were split at midline along the bolt path by a bandsaw and then underwent FRESH, CHILL24, CHILL48, and CHILL72 refrigeration treatments. The FRESH treatment involved images collected immediately after splitting each head, the CHILL24 treatment involved images collected after 24 h of refrigeration, the CHIL48 treatment involved images collected after 48 h of refrigeration, and the CHILL72 treatment involved images collected after 72 h of refrigeration. Measurements of soft tissue thickness, cranial thickness, total tissue thickness, and cross-sectional brain area were recorded for each refrigeration treatment. Soft tissue thickness did not differ caudal to (P = 0.3751) or rostral to (P = 0.2555) the bolt path. Cranial thickness did not differ caudal to (P = 0.9281) or rostral to (P = 0.9051) the bolt path. Total tissue thickness did not differ caudal to (P = 0.9225; FRESH: 24.77 mm, CHILL24: 23.93 mm, CHILL48: 24.27 mm, CHILL72: 42.30, SE: 0.86) or rostral to (P = 0.8931; FRESH: 24.09 mm, CHILL24: 23.99, CHILL48: 24.26, CHILL72: 24.43 mm, SE: 0.79 mm) the bolt path. Cross-sectional brain area was not different (P = 0.0971) between refrigeration treatments (FRESH: 9,829.65 ±â€…163.87 mm2, CHILL24: 10,012.00 ±â€…163.87 mm2, CHILL48: 9,672.43 ±â€…163.87 mm2, CHILL72: 10,235.00 ±â€…166.34 mm2). This study demonstrated that FRESH tissue parameters can be determined from cattle cadaver heads refrigerated for 24, 48, or 72 h.

Relationship of tissue dimensions and three captive bolt application sites on cadaver heads from mature swine (Sus scrofa domesticus) <200 kg body weight.

Penetrating captive bolt (PCB) is a common method of euthanasia for swine but has not been evaluated for mature swine < 200 kg body weight (BW). The objectives were to determine tissue depth, brain contact plane, and visible brain tissue damage (brain damage[BD]) for the common FRONTAL (F) and alternative TEMPORAL (T) and BEHIND EAR (BE) placements for PCB use on sows and boars weighing < 200 kg. Cadaver heads were obtained from 30 sows and 30 boars (estimated BW, mean ±â€…SD; sows: 165.8 ±â€…22.4 kg; boars: 173.6 ±â€…21.4 kg) from a slaughter establishment after electrical stunning and exsanguination. Heads were cooled at 2 to 4 °C for approximately 64 h. A Jarvis PAS-Type P 0.25R PCB with a Long Stunning Rod Nosepiece Assembly and a 3.5 GR power load was used for all PCB applications at the following placements: F-3.5 cm superior to the optic orbits at midline, T-at the depression posterior to the lateral canthus of the eye within the plane between the lateral canthus and the base of the ear, or BE-directly caudal to the pinna of the ear on the same plane as the eyes and targeting the middle of the opposite eye. For sows, the bolt path was in the brain for 10/10 (100.0%, 95% CI: 69.2% to 100.0%) F, T, and BE heads. In heads that could reliably be assessed for BD, BD was detected in 10/10 (100.0%, 95% CI: 69.2% to 100.0%) F heads, 9/9 (100.0%, 95% CI: 66.4% to 100.0%) T heads, and 0/10 (0.0%, 95% CI: 0.0% to 30.1%) BE heads. For boars, the bolt path was in the plane of the brain for 8/9 (88.9%, 95% CI: 51.8% to 99.7%) F heads, 9/10 (90.0%, 95% CI: 55.5% to 99.7%) T heads, and 11/11 (100.0%, 95% CI: 71.5% to 100.0%) BE heads. In heads that could reliably be assessed for BD, BD was detected in 8/9 (88.9%, 95% CI: 51.7% to 99.7%) F heads, 7/10 (70.0%, 95% CI: 34.8% to 93.3%) T heads, and 4/11 (36.4%, 95% CI: 10.9% to 69.2%) BE heads. Tissue depth was reported as mean ±â€…SE followed by 95% one-sided upper reference limit (URL). For sows, total tissue thickness differed (P < 0.05) between placements (F: 49.41 ±â€…2.74 mm, URL: 70.0 mm; T: 62.83 ±â€…1.83 mm, URL: 76.6 mm; BE: 84.63 ±â€…3.67 mm; URL: 112.3 mm). Total tissue thickness differed (P < 0.05) between placements for boars (F: 54.73 ±â€…3.23 mm, URL: 77.6 mm; T: 70.72 ±â€…3.60 mm, URL: 96.3 mm; BE: 92.81 ±â€…5.50 mm; URL: 135.3 mm). For swine between 120 and 200 kg BW, the F placement may have the greatest likelihood for successful euthanasia due to the least total tissue thickness and may present less risk for failure than the T and BE placements.

Euthanasia is an important procedure to protect animal welfare and cannot be avoided on swine farms. A common method of euthanasia for swine is penetrating captive bolt (PCB), which involves passing a metal bolt through the skull into the brain. This process should result in an immediate loss of consciousness. The use of PCB has been evaluated for market hogs and heavier sows and boars, but not for sows and boars weighing < 200 kg. As pigs mature, the cranial thickness at the common frontal PCB placement increases due to the expansion of sinus cavities. This makes PCB euthanasia more challenging for sows and boars. As a result, alternative PCB application sites, including behind the ear and at a temporal location, have been proposed. This study evaluated frontal, temporal, and behind-ear placements for PCB application to cadaver heads from sows and boars weighing < 200 kg. The frontal placement appeared to be more reliable than the temporal or behind ear placements due to the least tissue for the bolt to travel through to reach the brain, the greatest potential target area, and prevalent brain damage.

Evaluation of tissue depth, captive bolt penetration force and energy, and potential for bolt-thalamus contact in cadaver heads from physically castrated market barrows and immunocastrated boars.

The main objective of this study was to describe tissue thicknesses of cadaver heads from physically castrated market barrows (PC MARKET BARROWS) and immunocastrated boars (IC BOARS) at the frontal penetrating captive bolt (PCB) placement. Other objectives were to describe differences in bolt force and energy requirements to penetrate and describe potential for bolt-thalamus contact. Forty-four heads were obtained from PC MARKET BARROWS (n = 22) and IC BOARS (n = 22) of similar age and size that were rendered insensible with CO2. Mean HCW was 117.32 ±â€…3.52 kg. Snout to poll distance (cm) and maximum deflection distance (cm) were collected in duplicate. Heads were split at midline with a bandsaw and soft tissue and cranial thicknesses were measured with a digital caliper. Images of each cut surface were collected to evaluate the potential for thalamic damage. Tissue samples were retained from each half of each head and a universal tester was used to determine maximum force and energy of bolt penetration. There was no evidence to support a significant difference (P > 0.05) in tissue thicknesses between PC MARKET BARROWS and IC BOARS. Maximum deflection distance (maximum distance from a straight edge that was placed from the tip of the snout to the poll of the head) was not different (P = 0.10) between PC MARKET BARROWS (3.31 ±â€…0.10 cm) and IC BOARS (3.08 ±â€…0.10 cm). There was no evidence to support a difference (P = 0.77) in maximum force between PC MARKET BARROWS (7130.32 ±â€…483.23 N) and IC BOARS (6974.60 ±â€…463.70 N). There was also no evidence to support a difference (P = 0.62) in maximum energy between PC MARKET BARROWS (33.37 ±â€…2.77 J) and IC BOARS (32.04 ±â€…2.50 J). For PC MARKET BARROWS, there was a difference (P = 0.05) between the number of heads where the thalamus was located within the theoretical plane of bolt travel for market placement (21/21) versus mature placement (16/21). For IC BOARS, the number of heads where the thalamus was located within the plane of theoretical bolt path was not different between the two PCB placements (19/21 each). Overall, the data suggest that tissue profiles of PC MARKET BARROWS and IC BOARS do not differ at the frontal PCB placement site and the mechanical tools that are effective for PC MARKET BARROWS should also be effective for IC BOARS.

Broken tails in Holstein dairy cattle: A cross-sectional study.

Dairy cows are regularly handled when moved to the milking parlor and during other routine procedures. Low-stress handling methods are important in avoiding negative welfare states for dairy cattle. Tail twisting is used by some handlers to prompt cattle movement. However, when used inappropriately with excessive force, tail twisting can lead to a broken tail. The aim of this cross-sectional study was to determine cow-level factors that may be associated with the prevalence of broken tails in dairy cattle. A subset of 229 Holstein dairy cows (68 primiparous and 161 multiparous) at a single dairy were assessed for broken tails from the larger herd (N = 1,356). Tails were visually assessed for the presence of fractures by a single trained observer. A tail was classified as unfractured if it laid straight when at rest and as fractured if there were deviations in the tail when at rest. Poisson regression models were used to identify associations between cow-level characteristics and broken tails and compute adjusted prevalence ratios (PR). The prevalence of broken tails was 45.8% (105/229) at the time of assessment. Multiparous cows had a greater prevalence of broken tails than primiparous cows [PR = 1.70; 95% confidence interval (CI): 1.11-2.59]. The prevalence of broken tails was also greater for cows treated for mastitis ≥2 times than cows treated once for mastitis (PR = 1.84; 95% CI: 1.08-3.13) and cows never treated for mastitis (PR = 1.36; 95% CI: 1.02-1.82). Results from this study indicated that the longer a cow was present on the farm and the more times she was treated for mastitis, the more likely she was to experience a broken tail. These findings suggest that the relationship between dairy cow handling, health, and welfare is a multifactorial issue.

Assessment of United States Department of Agriculture Food Safety Inspection Service Humane Handling Enforcement Actions: 2018-2020.

Federally inspected slaughter establishments in the United States must adhere to the Humane Methods of Slaughter Act and regulations that enforce it. Failure to comply with this law results in a Humane Handling Enforcement Action (HHEA) issued by the United States Department of Agriculture Food Safety Inspection Service (USDA FSIS). The objective of this study was to systematically analyze and describe HHEAs issued between 2018 and 2020. Enforcement action notification letters were accessed from the USDA FSIS website and date, location, regulatory action, reason for noncompliance, species, and follow up action for each HHEA was recorded. Summary statistics (proportions and percentages) were calculated for the entire population dataset. Between 2018 and 2020, FSIS issued 293 HHEAs; 109 in 2018, 85 in 2019, and 99 in 2020. The majority of HHEAs (64.16%; 188 of 293) were related to the mechanical stunning of bovine (39.93%; 117 of 293) and porcine (24.23%; 71 of 293) species. The majority (50.23%; 107 of 213) of causative reasons for mechanical stun failure across all species were not clearly described; however, of those that were, most (39.12%; 68 of 213) were related to the placement of mechanical stuns. Addressing these issues through improved training and research would help to reduce the total number of HHEAs. Additional detail in reporting the events that result in HHEAs from USDA FSIS would aid in guiding corrective actions on an industry-wide scale.

History and best practices of captive bolt euthanasia for swine.

The definition of animal welfare includes how an animal dies. As such, euthanasia is intrinsically linked to animal welfare, and ensuring a good death through effective, safe, and validated practices is a critical piece of promoting positive animal welfare. The objective of this review is to provide a better understanding of the literature on the euthanasia of swine via penetrating captive bolt (PCB) and nonpenetrating captive bolt (NPCB), as well as a history of captive bolt use, and indicators of sensibility and insensibility. To do this, we performed a systematic review that included 30 peer-reviewed articles and 17 other publications. NPCB devices have been validated as an effective single-step euthanasia method for neonatal and preweaning swine, as well as a two-step euthanasia method for nursery swine. PCB devices have been validated as an effective euthanasia method for nursery and market swine up to 120 kg, but further investigation is required for the use of captive bolt devices on mature breeding sows and boars.

Relationship of tissue dimensions and three captive bolt placements on cadaver heads from mature swine (Sus scrofa domesticus) > 200 kg body weight.

Three penetrating captive bolt (PCB) placements were tested on cadaver heads from swine with estimated body weight (BW) >200 kg (sows = 232.9 ± 4.1 kg; boars = 229.3 ± 2.6 kg). The objectives were to determine tissue depth, cross-sectional brain area, visible brain damage (BD), regions of BD, and bolt-brain contact; and determine relationships between external head dimensions and tissue depth at each placement. A Jarvis PAS-Type P 0.25R PCB with a Long Stunning Rod Nosepiece Assembly and 3.5 g power loads was used at the following placements on heads from 111 sows and 46 boars after storage at 2 to 4 °C for ~62 h before treatment: FRONTAL (F)-3.5 cm superior to the optic orbits at midline, TEMPORAL (T)-at the depression posterior to the lateral canthus of the eye within the plane between the lateral canthus and the base of the ear, or BEHIND EAR (BE)-directly caudal to the pinna of the ear on the same plane as the eyes and targeting the middle of the opposite eye. For sows, the bolt path was in the plane of the brain for 42/42 (100%, 95% confidence interval [CI]: 91.6% to 100.0%) F heads, 39/40 (97.5%, 95% CI: 86.8% to 99.9%) T heads, and 34/39 (87.5%, 95% CI: 72.6% to 95.7%) BE heads; for the heads that could reliably be assessed for BD damage was detected in 25/26 (96.2%, 95% CI: 80.4% to 99.9%) F heads, 24/35 (68.6%, 95% CI: 50.7% to 83.2%) T heads, and 5/40 (12.5%, 95% CI: 4.2% to 26.8%) BE heads. For boars, the bolt path was in the plane of the brain for 17/17 (100.0%, 95% CI: 80.5% to 100.0%) F heads, 18/18 (100.0%, 95% CI: 81.5% to 100.0%) T heads, and 14/14 (100.0%, 95% CI: 76.8% to 100.0%) BE heads; damage was detected in 11/12 (91.7%, 95% CI: 61.5% to 99.8%) F heads, 2/15 (13.3%, 95% CI: 1.7% to 40.5%) T heads, and 7/14 (50.0%, 95% CI: 23.0% to 77.0%) BE heads. Tissue depth was reported as mean ± standard error followed by 95% one-sided upper reference limit (URL). For sows, total tissue thickness was different (P < 0.05) between placements (F: 52.7 ± 1.0 mm, URL: 64.1 mm; T: 69.8 ± 1.4 mm, URL: 83.9 mm; BE: 89.3 ± 1.5 mm, URL: 103.4 mm). In boars, total tissue thickness was different (P < 0.05) between placements (F: 41.2 ± 2.1 mm, URL: 56.3 mm; T: 73.2 ± 1.5 mm, URL: 83.4 mm; BE: 90.9 ± 3.5 mm, URL: 113.5 mm). For swine > 200 kg BW, F placement may be more effective than T or BE due to less soft tissue thickness, which may reduce concussive force. The brain was within the plane of bolt travel for 100% of F heads with BD for 96.2% and 91.7% of F sow and boar heads, respectively.

Efficacy of oral meloxicam as primary pain mitigation following caustic paste disbudding of three day old Holstein calves1.

The purpose of this study was to investigate the effects of perioperative administration of oral meloxicam prior to and following the application of caustic paste to disbud neonatal dairy calves. Sixty-one 3-4-d-old Holstein heifer calves were randomly assigned to one of four treatment groups of 15-16 calves. The treatment groups were: 1) M1, caustic paste disbudding and oral meloxicam (45 mg) with a placebo 24 h later; 2) M2, treatment M1 followed by a second 45-mg dose of meloxicam 24 h later instead of placebo; 3) CONTROL, treatment M1 with placebo in place of meloxicam; and 4) SHAM, sham disbudding with placebo in place of meloxicam. Infrared thermography was used to quantify eye and horn bud temperatures. Pressure algometry was used to measure Mechanical nociceptive threshold (MNT) surrounding the horn bud. Average daily gain and body weight (BW) were obtained by weighing each animal throughout the study and calculating the changes over time. Plasma was collected and analyzed for cortisol and substance P concentrations. Substance P and cortisol decreased in all animals over time, regardless of treatment. Mean plasma substance P concentration across all time points was greater (P < 0.05) in the SHAM group than M1 or M2 but not different (P > 0.05) than the CONTROL group. The MNT and ocular temperatures decreased over time across all treatments (P < 0.05). Mean BW increased over time across all treatments (P < 0.05). A significant interaction (P < 0.05) between treatment and sampling time was observed at 12 h following treatment application for both mean horn bud temperature and the ratio between horn bud and ocular temperature. Overall, the results of this study suggest that meloxicam administration at a dose of 45 mg per animal may have limited influence as the primary modulator of pain and inflammatory response in calves that have been disbudded with caustic paste at 3 d of age.

Quantification of cooling effects on basic tissue measurements and exposed cross-sectional brain area of cadaver heads from market pigs.

The objective of this project was to determine the impact of cooling on the soft tissue thickness, cranial thickness, and cross-sectional brain area of cadaver heads from market pigs. Documenting the effect of cooling on tissue dimensions of swine heads is valuable and important for future investigations of physical stunning and euthanasia methods that use cadaver heads. Scalded and dehaired cadaver heads with intact jowls were sourced from market pigs stunned with CO2 gas. After transport to the data collection location, a penetrating captive bolt (PCB) shot (Jarvis Model PAS-Type P 0.25R Caliber Captive Bolt Pistol with Medium Rod Assembly and Blue Powder Cartridges) was applied in the frontal position. Following PCB application, each head (n = 36) underwent an UNCHILLED treatment followed by CHILLED treatment. The UNCHILLED treatment involved images collected immediately after splitting each head along the bolt path, and the CHILLED treatment involved images of the same heads after storage in a walk-in cooler for 24 h at 2 to 4°C. All measurements for each treatment were collected from images of the heads on the plane of the bolt path immediately prior to and immediately after the refrigeration treatment. Measurements were performed by two observers. Across all measurements, mean interobserver coefficient of variation was 11.3 ± 0.6%. The soft tissue caudal to the bolt path was different (P = 0.0120) between treatments (CHILLED: 6.4 ± 0.2 mm; UNCHILLED: 7.2 ± 0.2 mm). The soft tissue thickness rostral to the bolt path was different (P = 0.0378) between treatments (CHILLED: 5.5 ± 0.2 mm; UNCHILLED: 6.1 ± 0.2 mm). Cranial thickness caudal to the bolt path was not different (P = 0.8659; CHILLED: 18.1 ± 0.6 mm; UNCHILLED: 18.3 ± 0.6 mm), nor was there a significant difference (P = 0.2593) in cranial thickness rostral to the bolt path between treatments (CHILLED: 16.2 ± 0.6 mm; UNCHILLED: 15.2 ± 0.6 mm). Cross-sectional brain area did not differ (P = 0.0737; CHILLED: 3633.4 ± 44.1 mm; UNCHILLED: 3519.9 ± 44.1 mm). A correction factor of 1.12 was determined from this study for cases where estimation of UNCHILLED soft tissue thickness from CHILLED soft tissue thickness is necessary.

Relationship of captive bolt stunning location with basic tissue measurements and exposed cross-sectional brain area in cadaver heads from market pigs.

The objective of this study was to contrast the soft tissue thickness, cranial thickness, total tissue thickness, cross-sectional brain area, and bolt-brain contact from the common frontal application of captive bolt euthanasia with the alternative location behind the ear in cadaver swine heads. Twenty-three cadaver heads from pigs that were approximately 136 kg and 6 mo of age were collected from a regional slaughter establishment following CO2 stunning and assigned to either the FRONTAL (n = 11) or the CAUDAL TO PINNA (n = 12) application of the captive bolt. The soft tissue thickness was different (P < 0.0001) between the 2 applications (FRONTAL: 8.3 ± 3.4 mm; CAUDAL TO PINNA: 56.5 ± 3.4 mm). The cranial thickness was different (P < 0.0001) between the applications (FRONTAL: 23.4 ± 2.9 mm; CAUDAL TO PINNA: 26.5 ± 2.9 mm). There was also a difference (P < 0.0001) in the total tissue thickness between the 2 applications (FRONTAL: 31.7 ± 3.8 mm; CAUDAL TO PINNA: 73.4 ± 3.8 mm). Cross-sectional area was calculated from images collected immediately after the heads were cut along the plane of bolt travel by bandsaw and was different (P = 0.0028) between the 2 applications (FRONTAL: 25.2 ± 1.3 cm2; CAUDAL TO PINNA: 18.9 ± 1.3 cm2). Bolt-brain contact was also assessed from the images, and a difference (P = 0.0360) between the 2 applications (FRONTAL: 100 ± 10.5%; CAUDAL TO PINNA: 66.7 ± 10.5%) was identified. The results of this study suggest that the FRONTAL application may provide a bolt path with less tissue to travel through when compared with the CAUDAL TO PINNA application for pigs of the approximate age and weight of those in this study. Ultimately, the FRONTAL location may present less risk for the captive bolt euthanasia of swine at market weight at this time. Additional refinement of the CAUDAL TO PINNA procedure and modification to the captive bolt device to penetrate to a suitable depth to ensure brain damage is recommended.

Injection order effects on efficacy of calcium chloride and sodium tripolyphosphate in controlling the pink color defect in uncured, intact turkey breast.

An experiment was conducted to test sequential injection of sodium tripolyphosphate (STP; 0.5% meat weight basis, mwb) followed by injection with or without addition of calcium chloride (CaCl(2), 500 ppm mwb), and to test the effect of post-injection delay prior to cooking. A second experiment evaluated the impact of injection order and delay time between independent addition of CaCl(2) (500 ppm mwb) and STP (0.5% mwb). Turkey was formulated without an added pink generating ligand (NONE), with nicotinamide (NIC; 0.1% mwb), or with sodium nitrite (NIT; 10 ppm mwb). A white colloid was observed in the extracellular space of treatments containing both STP and CaCl(2.) Addition of CaCl(2) decreased nitrosylhemochrome but did not reduce levels of nicotinamide hemochrome or CIE a(*) values. Injection order or delay between injections did not contribute to controlling the pink defect in cooked, intact turkey breast.