Rapid Establishment of a Cold Chain Capacity of -60°C or Colder for the STRIVE Ebola Vaccine Trial During the Ebola Outbreak in Sierra Leone.

Clinical Trials Registration: ClinicalTrials.gov [NCT02378753] and Pan African Clinical Trials Registry [PACTR201502001037220].

The effect of extended refrigerated storage on the physicochemical, structural, and microbial quality of sous vide cooked biceps femoris treated with ginger powder (zingibain).

The aim of the study was to investigate quality and shelf life of beef meat cooked under sous vide conditions then extended refrigerated storage for 10 weeks. Biceps femoris (n = 6) from six to seven year old cows were treated with 2 g/L ginger powder (GP) containing zingibain or control (no injection) and were then cooked in sous vide conditions at 65 °C for 1 h or 8 h. Cooked samples were evaluated for physicochemical (pH, total water content, cooking loss, Warner-Bratzler shear force (WBSF), texture profile analysis (TPA), L*, a*, b* properties and thiobarbituric acid reactive substance (TBARS)), microstructure (scanning electron microscopy) and microbiological (Brochothrix thermospacta, Clostridium perfringens, Lactic acid bacteria, Listeria monocytogenes, Salmonella spp, and yeasts and moulds) quality after vacuum packing, cooking, then refrigerated storage at 4 °C for 0, 2, 4, 8 or 10 weeks. Physicochemical parameters were improved by GP treatment (P < 0.05) while there was no effect of storage time on WBSF, TPA or microstructure. The microbial quality of sous vide cooked meat in refrigerated storage appeared to be four weeks and oxidation shelf life of the cooked meat was found to be two weeks under refrigerated storage.

A microfluidic device and instrument prototypes for the detection of Escherichia coli in water samples using a phage-based bioluminescence assay.

Current quantification methods of Escherichia coli (E. coli) contamination in water samples involve long incubation, laboratory equipment and facilities, or complex processes that require specialized training for accurate operation and interpretation. To address these limitations, we have developed a microfluidic device and portable instrument prototypes capable of performing a rapid and highly sensitive bacteriophage-based assay to detect E. coli cells with detection limit comparable to traditional methods in a fraction of the time. The microfluidic device combines membrane filtration and selective enrichment using T7-NanoLuc-CBM, a genetically engineered bacteriophage, to identify 4.1 E. coli CFU in 100 mL of drinking water within 5.5 hours. The microfluidic device was designed and tested to process up to 100 mL of real-world drinking water samples with turbidities below 10 NTU. Prototypes of custom instrumentation, compatible with our valveless microfluidic device and capable of performing all of the assay's units of operation with minimal user intervention, demonstrated similar assay performance to that obtained on the benchtop assay. This research is the first step towards a faster, portable, and semi-automated, phage-based microfluidic platform for improved in-field water quality monitoring in low-resource settings.

Improving tenderness and quality of M. biceps femoris from older cows through concentrate feeding, zingibain protease and sous vide cooking.

The study investigated the effect of zingibain protease and sous vide cooking on tenderness and water-holding capacity of M. biceps femoris (BF) from 30 older Angus cows (6-7 years) fed concentrates for 0, 28, 42 or 56 days. BF were cooked for 1, 8, and 18 h at 65 °C and 75 °C, without any pre-treatment, after they have been injected with water only or after they have been injected with either 1 g/L or 2 g/L ginger powder solution (containing zingibain). Samples were tested for cooking loss, total water content, Warner-Bratzler shear force (WBSF), collagen content, and myofibrillar fragmentation index (MFI). Results revealed the significant interactions between concentrate feeding, ginger powder injection, cooking temperature, and time on quality traits. WBSF was reduced (P < 0.001) by increasing zingibain concentration at 65 °C but a gradual decrease was noted at 75 °C. Collagen solubility and MFI increased (P < 0.05) with increasing zingibain concentration. Injecting zingibain along with sous vide cooking demonstrated the weakening of myofibrillar and connective tissue proteins contributing to enhanced collagen solubility and tenderness in BF.

Effect of sous vide cooking and ageing on tenderness and water-holding capacity of low-value beef muscles from young and older animals.

This study investigated the effect of the age of the animal, sous vide cooking and ageing on tenderness and water-holding capacity of bovine biceps femoris (BF) and semitendinosus (ST). Samples of each muscle from young (<18 months) and older (30-42 months) animals, at 0 and 13 days ageing, were cooked at 55 °C, 65 °C, and 75 °C for 1 h, 8 h and 18 h and tested for Warner-Bratzler shear force (WBSF), cooking loss, total water content and collagen solubility. WBSF reduced with ageing (P < 0.05) and sous vide cooking (P < 0.001) in both muscles. Our results demonstrated that meat from older animals required a higher temperature (75 °C) and prolonged cooking (18 h) to achieve equal tenderness in both BF and ST relative to young animals. Cooking time, temperature and their interaction altered cooking loss (P < 0.001) for both muscles. The higher cooking temperature increased collagen solubility (P < 0.001) in both muscles and solubilisation of collagen may have contributed to improved tenderness of BF and ST in sous vide cooking.

Sensory and Physical Characteristics of M. biceps femoris from Older Cows Using Ginger Powder (Zingibain) and Sous Vide Cooking.

This study aimed to evaluate the sensory and physical characteristics of zingibain-injected meat combined with sous vide cooking. M. biceps femoris (BF; n = 12) acquired from 6-7 year old Angus cows were cooked using the sous vide method at 65 °C, for 8 h or 12 h, either with ginger powder (GP) injected in a 2 g/L solution in water (treatment) or un-injected (control). The sensory attributes included flavour, juiciness, tenderness, and physicochemical characteristics were Warner-Bratzler shear (WBSF), hardness, total water content (TWC), cooking loss (CL) and collagen content. A significant improvement in tenderness with injection treatment and cooking time was observed, as evaluated through trained sensory panellists, and reduced WBSF and hardness (p < 0.05 for all). The flavour of the meat was not affected by injection treatment or cooking time (p > 0.05), but juiciness and TWC were reduced with longer cooking times (p < 0.01 for both). Soluble collagen increased with injection treatment and cooking time (both p < 0.05). Moderate to high correlations were found between sensory and physical measurements for tenderness and juiciness. The longer cooking time (12 h) with GP injection treatment caused over tenderization of the meat. The soft texture associated with over-tenderization may be suitable for some specialised consumer markets, for instance, the elderly population with chewing difficulties. Improving the eating quality of low-quality meat from old animals through sous vide cooking and the use of ginger proteases may increase the acceptability of lower value beef, potentially enhancing the commercial value of carcasses typically produced in the beef industry.

Maternal supplementation of twin bearing ewes with calcium and magnesium alters immune status and weight gain of their lambs.

This study investigated the effect of supplementation of ewes with calcium (Ca) and magnesium (Mg) in late gestation and early lactation on the plasma mineral concentration, hormone profile and immunity responses of ewes and their lambs. Twin bearing ewes were allocated between four dietary treatment groups (n = 11 per treatment): control (0.33% DM Ca and 0.28% DM Mg); high Ca (0.72% DM Ca and 0.28% DM Mg); high Mg (0.33% DM Ca, 0.48% DM Mg); and high Ca+Mg (0.66% DM Ca and 0.47% DM Mg), offered as part of a whole ration. Ewes were fed the treatment supplement from around one month prior to lambing to one month after lactation. Blood and urine samples were collected at seven time-points: weekly starting at 5 weeks prior to lambing; within 12 h post lambing (+12 h); and then at fortnightly intervals at 2 week (+2 W) and at 4 week (+4 W) post lambing. Colostrum/milk samples from ewes and blood samples from lambs were collected at +12 h, +2 W and +4 W. Live weight of lambs were measured at +12 h, +2 W and +4 W. The plasma concentration of PTH, 1,25(OH)2D3 and 25(OH)D3 was lowest at +2 W (P ≤ 0.002). Ewes from the Ca+Mg group had the lowest mean concentration of 1,25(OH)2D3 than the other groups (P = 0.005). Magnesium supplementation improved the plasma Mg concentration over time in ewes (P <  0.001) and lambs from the control group had lower plasma Mg concentration compared to the treatment groups at +4 W (P = 0.001). Oxidative burst response in lambs supplemented with Ca tended to be greater at +4 W than the other groups at the same time point (P = 0.051) and Mg supplementation increased total antioxidant capacity (TAC) concentration in lambs (P = 0.040). The average daily weight gain of lambs was 204 g/lamb/d for the Ca group, 207 g/lamb/d for the Mg group, 245 g/lamb/d for the Ca+Mg group which were greater than the control group (148 g/lamb/d) (P < 0.001). Despite the normal concentration of Ca and Mg in the plasma, supplementation of ewes with Ca and Mg from one month prior to lambing to one month post lambing improved TAC concentration and weight gain in lambs.

A Survey of New South Wales Sheep Producer Practices and Perceptions on Lamb Mortality and Ewe Supplementation.

High lamb mortality rates reduce profitability and reduce the perceived animal welfare standards of the industry. This study aimed to understand producer knowledge of lamb mortality rates and causes of lamb mortality, and to investigate various practices and perceptions of producers that may contribute to lamb deaths. Postal and online surveys gathered data on Australian sheep producer's knowledge and practices around lambing and management practices. Based on results, approximately 50% of producers estimated less than 10% mortality of lambs between birth and marking, compared to published data estimates of 20-25% mortality. Pre-lambing vaccination of ewes was not undertaken by 10-20% of producers. Ninety-six percent of producers vaccinated lambs; however, 17% of Merino and 23% of crossbred lamb producers only gave a single vaccination instead of the recommended initial vaccine and booster. The lower estimated mortality impacts producer's perceived benefits of management strategies being undertaken. Research undertaken needs to be more effectively distributed to producers via extension services to ensure producers understand the causes of mortality. Important messages to convey to producers include the limited impact of predation in most cases and the total costs of lamb mortality on-farm.

A Robust Incubator to Improve Access to Microbiological Culture in Low Resource Environments.

To help address the limitations of operating conventional microbiological culture incubators in low resource environments, a new incubator design was developed and tested to meet the requirements of operation in laboratories without reliable power (power outages up to 12 contiguous hours) or climate control (ambient indoor temperatures from 5 °C to 45 °C). The device is designed to enable adherence to incubation temperatures recommended for growth detection, identification, and drug susceptibility testing (DST) of human pathogenic bacteria. During power outages, stable temperatures are maintained in the device's internal sample compartment by employing phase change material (PCM) as a bi-directional thermal battery to maintain incubation temperature. Five prototypes were tested in a laboratory setting using environmental test chambers and programmable power supplies, and three were field tested in the Lao PDR in situations of intended use. The prototypes successfully held their temperature to within ±1 °C in both laboratory environmental chamber testing as well as during the field test. The results indicate that the device will maintain stable culture temperatures across periods of intermittent power supply, while enabling normal workflow of this could greatly increase the availability of microbiological culture for diagnosis and antimicrobial resistance (AMR) monitoring.

Effects of chilled and frozen storage conditions on the lamb M. longissimus lumborum fatty acid and lipid oxidation parameters.

This study investigated the effects of chilled and frozen storage conditions on the fatty acid profile and lipid oxidation in lamb M. longissimus lumborum (LL). Muscle LL (n=360) were randomly selected at 24h post-mortem from a commercial Australian abattoir and maintained chilled for five periods (0, 2, 4, 6 and 8weeks) followed by frozen for six periods (0, 4, 8, 12, 24 and 52weeks) at -12°C or -18°C. Samples were analysed for fatty acid profile and lipid oxidative parameters (TBARS, oxidation-reduction potential and peroxidase activity). Health-claimable polyunsaturated fatty acids (EPA and DHA) were unaffected by neither storage period nor temperature. TBARS levels did not exceed rancidity thresholds recommended in the literature, with these and other lipid peroxidation parameters showing broad increases in concentration with more 'long-term' chilled and frozen storage periods. Combined, the results suggest nutritional and eating quality were maintained across the various chilled and frozen storage conditions applied in this study.

Effects of chilled-then-frozen storage (up to 52weeks) on an indicator of protein oxidation and indices of protein degradation in lamb M. longissimus lumborum.

This study investigated the protein oxidation properties of lamb following chilled-then-frozen storage. Experimental (n=360) M. longissimus lumborum (LL) were randomly sampled from the boning room of a commercial Australian abattoir, at 24h post mortem, and assigned to five chilled storage periods (0, 2, 4, 6 and 8weeks) and six subsequent frozen storage periods (0, 4, 8, 12, 24 and 52weeks). Upon completion of each storage treatment combination, corresponding LL were sub-sectioned and analysed for carbonyl content, protein solubility, nitrate/nitrite content, particle size analysis and estimated myoglobin fractions. The association between these protein measures and shear force was also explored. During chilled storage, particle size and sarcoplasmic protein solubility decreased which indicated protein degradation, while frozen storage only affected myoglobin oxidation. Tenderness was best explained by decreased particle size, decreased deoxymyoglobin and increased oxymyoglobin. No carbonyl effects were observed. It can be concluded that, according to these analyses, that in chilled-then-frozen lamb carbonyl formation was negligible.

Effects of chilled-then-frozen storage (up to 52weeks) on lamb M. longissimus lumborum quality and safety parameters.

This study evaluated the effect of chilled followed by frozen storage on lamb quality and safety parameters. Experimental (n=360) M. longissimus lumborum (LL) were randomly sampled from the boning room of a commercial Australian abattoir, at 24 h post-mortem, and assigned to five chilled storage periods (0, 2, 4, 6 and 8 weeks) and six subsequent frozen storage periods (0, 4, 8, 12, 24 and 52 weeks). Upon completion of each storage treatment combination, corresponding LL were sub-sectioned and analysed for colour stability (0, 1, 2 and 3 days), shear force, fluid losses (purge, thaw and cooking losses), intramuscular fat content, sarcomere length, water activity and microbial load (lactic acid bacteria, Enterobacteriaceae sp., Brochothrix thermosphacta, Clostridium perfringens and Escherichia coli). LL stored chilled for 2-4 weeks prior to freezing presented superior results for shear force, display colour and low levels of spoilage microbes, correlating with good eating quality and safety following more than one year of frozen storage.

Long-term red meat preservation using chilled and frozen storage combinations: A review.

This paper reviews current literature relating to the effects of chilled and frozen storage on the quality characteristics of red meat (lamb and beef). These characteristics include tenderness (shear force), juiciness (fluid losses), flavour (lipid oxidation), colour and microbial loading. Research suggests that chilled storage of red meat can improve certain properties, such as shear force and species-specific flavour, to threshold levels before the effects on flavour and colour become deleterious, and key microbial species proliferate to spoilage levels. For frozen red meat, the negative effects upon quality traits are prevented when stored for shorter durations, although chilled storage conditions prior to freezing and retail display post-thawing can both positively and negatively affect these traits. Here, we review the effects of different chilled, frozen and combined chilled and frozen storage practices (particularly the chilled-then-frozen combination) on meat quality and spoilage traits, in order to contribute to superior management of these traits during product distribution.

The effect of forage-types on the fatty acid profile, lipid and protein oxidation, and retail colour stability of muscles from White Dorper lambs.

The aim of this study was to evaluate the effect of different forage-types on lamb meat quality parameters. White Dorper lambs that had grazed five forage-types, were slaughtered commercially. At 24h post-mortem, the m. longissimus lumborum (LL) was removed from one side, sliced into three equal sub-samples, vacuum packaged and assigned to ageing periods (5, 12 or 40days); the other side of LL was aged for 5days. The m. adductor femoris was used for fatty acid analysis. Lambs fed chicory+arrowleaf clover had the highest concentration of health claimable omega-3 fatty acids and the lowest omega-6:omega-3 fatty acid ratio. Forage-types with higher vitamin E content showed lower lipid oxidation levels independent of ageing period. Forage-type and ageing period did not influence the redness, yellowness, chroma or reflectance ratio (630nm÷580nm) of displayed meat. Chicory+arrowleaf clover gave the best results to improve the fatty acid content of lamb meat.

The effect of forage type on lamb carcass traits, meat quality and sensory traits.

The aim of this study was to evaluate the effect of different forage-types on lamb carcass, meat quality and sensory attributes. Sixty-two, White Dorper lambs finished on bladder clover, brassica, chicory+arrowleaf clover, lucerne+phalaris or lucerne, were slaughtered at a commercial abattoir. At 24h post-mortem, the m. longissimus thoracis et lumborum (LL) was removed from the left side and sliced into three equal sub-samples, vacuum packaged and randomly assigned to ageing periods (5, 12 or 40days) and the right side was aged for 5days. The m. semimembranosus and m. adductor femoris were removed and, the former was then aged for 40days. Lambs fed chicory+arrowleaf clover or lucerne had a higher dressing percentage and fat depth. Bladder clover gave the highest level of glycogen in the LL. No sensory or other meat quality trait differences were found between the treatments. In general, no treatments showed any unfavourable effect on the traits examined.

Differential response to an algae supplement high in DHA mediated by maternal periconceptional diet: intergenerational effects of n-6 fatty acids.

Algae high in docosahexaenoic acid (DHA) may provide a source of long-chain omega-3 polyunsaturated fatty acids (LCn-3PUFA) for inclusion in the diet of lambs to improve the LCn-3PUFA status of meat. The effect of background LCn-3PUFA status on the metabolism of high DHA algae is, however, unknown. The aim of the current study was to determine whether the response to a high in DHA algae supplement fed to lambs for six weeks prior to slaughter was mediated by a maternal periconceptional diet. Forty Poll Dorset × Border Leicester × Merino weaner lambs were allocated to receive either a ration based on oat grain, lupin grain, and chopped lucerne (control) or the control ration with DHA-Gold™ algae included at 1.92 % DM (Algae) based on whether the dams of lambs had previously been fed a diet high in n-3 or n-6 around conception. LCn-3PUFA concentration was determined in plasma and red blood cells (RBC) prior to and following feeding. The concentrations of EPA and DHA in the plasma and RBC of lambs receiving the control ration were significantly (p < 0.001) lower when lambs received the ration for 14 days compared with pre-feeding concentrations. The concentrations of EPA and DHA were also significantly (p < 0.001) higher when lambs consumed the Algae ration compared with the control ration for 42 days. The increase in EPA and DHA was, however, significantly (p < 0.05) lower if lamb dams had previously been fed a diet high in n-6 at conception. Assessing the previous nutrition and n-3 status of lambs may allow producers to more accurately predict the likely response to supplements high in LCn-3PUFA, particularly, DHA.

Increased prostaglandin response to oxytocin in ewes fed a diet high in omega-6 polyunsaturated fatty acids.

Diets high in omega-6 polyunsaturated fatty acids (n-6) are associated with increased prostaglandin F(2α) (PGF(2α)) synthesis in cattle, however, the specific effects on the potential prostaglandin response to an oxytocin challenge in sheep have not been reported. The aim of the current study was to determine whether oxytocin-stimulated PGF(2α) was significantly increased when ewes were fed a diet high in n-6 compared with a control diet low in n-6. Merino x Border Leicester ewes (n = 30) received one of two dietary treatments, either high in n-6 (70 % oat grain) or low in n-6 (control diet, 100 % cereal/legume silage). Ewes consumed the diets for 44 days prior to two consecutive oxytocin challenges. Plasma n-6 and PGF(2α) metabolite (PGFM) concentrations following oxytocin challenge were greater (P < 0.05) when ewes were fed a diet high in n-6 compared with the control diet. A higher availability of n-6 may have lead to an increased in vivo synthesis of PGF(2α), however, further research is required to determine the exact mechanisms involved.

Pressure ulcer prevalence and risk factors during prolonged surgical procedures.

Pressure ulcer formation related to positioning while in the OR increases the length of hospital stay and hospital costs, but there is little evidence documenting how positioning devices used in the OR influence pressure ulcer development when examined with traditional risk factors. The aim of this prospective cohort study was to identify the prevalence of and risk factors associated with pressure ulcer development among patients undergoing surgical procedures lasting longer than three hours. Participants included all adult same-day admission patients scheduled for a three-hour surgical procedure during an eight-month period (N = 258). Data were gathered preoperatively, intraoperatively, and postoperatively on pressure ulcer risk factors. Bivariate analyses indicated that the type of positioning (ie, heels elevated) (χ(2) = 7.897, P = .048), OR bed surface (ie, foam table pad) (χ(2) = 15.848, P = .000), skin assessment in the postanesthesia care unit (χ(2) = 41.652, P = .000), and male gender (χ(2) = 6.984, P = .030) were associated with pressure ulcer development. Logistic regression analyses indicated that the use of a foam pad (ß = 2.691, P = .024) and a lower day-one Braden score (ß = .244, P = .003) were predictive of pressure ulcers.