The use of cells frozen at -196o for the bioassay of preservatives

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Polpa de jambolão (Eugenia jambolana Lam.) fresca e desidratada: características físico-químicas, bioativas e funcionais, efeitos biológicos em Caenorhabditis elegans e uso para produção de frozen yogurt caprino probiótico

This work evaluated the fresh, spray dried (with 10 % of Arabic Gum) and freeze dried jambolan pulp (Eugenia jambolana Lam.) in regard to physicochemical (pH, moisture, water activity, average particle diameter, solubility and color), bioactive [total phenolic content (TPC), monomeric anthocyanin, pronathocyanidin (PA), total elagic acid (TEA), myricetin and cyanidin] and in vitro functionality (antioxidant, antienzymatic and antimicrobial activities]. In addition, the in vivo functionality of jambolan pulp was investigated using the Caenorhabditis elegans model for insulin signaling, longevity and induced neurodegeneration (Alzheimer’s disease and Parkinson’s disease related symptoms). The dried jambolan pulp presented TPC retention (50% to 75%), PA (90% to 98%), TEA (31% to 83%), myricetin (40% to 84%), cyanidin (72% to 84%) and antioxidant activity (15%). The fresh jambolan pulp, the freeze dried pulp and the spray dried jambolan pulp presented high enzymatic inhibitory activity against pancreatic lipase (4,4 to 5,8 mg/mL), alpha-glycosidase (10,3 to 13,8 mg/mL) and alpha-amylase (8,9 to 11,2 mg/mL). They also were active inhibitors against the pathogen S. aureus. The dried jambolan experimental samples were able to increase the expression of several genes linked to the insulin signaling pathways (SIR-2.1, PPTR-1, DAF-16, SOD-3, e CTL) and increased the lifespan in C. elegans (18,07 % - 24,34 %), besides decreasing the amyloid AB1-42 aggregation induced paralysis and MPP+ (1-methyl-4-phenylpyridinium) induced neurodegeneration. Based on that, the jambolan pulp and the spray dried jambolan pulp were further selected for the production of caprine frozen yogurt with the addition of Bifidobacterium animalis subsp. lactis BI-07. The final product were evaluated in regard to their physicochemical (pH, acidity, total solids, protein, total reducing sugars, fat, ashes, overrun, melting test), bioactive (TPC and monomeric anthocyanin, antioxidant activity, probiotic viability and sensory analysis (sensory acceptance). The results showed that samples with probiotic had lowest pH and higher acidity, TPC, anthocyanin and antioxidant activity. It was also observed low overrun (14.2% to 22.6%). vi Samples with probiotic had lower flavor scores. Overall, this research presents the jambolan as a highly functional bioactive-rich fruit with the potential to modulate important biological pathways, extend lifespan and retard the development of neurodegenerative diseases. Jambolan is an underexploited exotic fruit with a high colorant potential and this thesis shows for the first time in the literature important technological, biological and scientific data about this fruit that could be used towards the development of health-oriented food products.

A metagenomic comparison of endemic viruses from broiler chickens with runting stunting syndrome and from normal birds

Runting-stunting syndrome (RSS) in broiler chickens is an enteric disease that causes significant economic losses to poultry producers worldwide due to elevated feed conversion ratios, decreased body weight during growth, and excessive culling. Of specific interest are the viral agents associated with RSS which have been difficult to fully characterise to date. Past research into the aetiology of RSS has implicated a wide variety of RNA and DNA viruses however, to date, no individual virus has been identified as the main agent of RSS and the current opinion is that it may be caused by a community of viruses, collectively known as the virome. This paper attempts to characterise the viral pathogens associated with 2 – 3 week old RSS-affected and unaffected broiler chickens using next-generation sequencing and comparative metagenomics. Analysis of the viromes identified a total of 20 DNA & RNA viral families, along with 2 unidentified categories, comprised of 31 distinct viral genera and 7 unclassified genera. The most abundant viral families identified in this study were the Astroviridae, Caliciviridae, Picornaviridae, Parvoviridae, Coronaviridae, Siphoviridae, and Myoviridae. This study has identified historically significant viruses associated with the disease such as chicken astrovirus, avian nephritis virus, chicken parvovirus, and chicken calicivirus along with relatively novel viruses such as chicken megrivirus and sicinivirus 1 and will help expand the knowledge related to enteric disease in broiler chickens, provide insights into the viral constituents of a healthy avian gut, and identify a variety of enteric viruses and viral communities appropriate for further study.

Astaxanthin profiles and corresponding colour properties in Australian farmed black tiger prawn (Penaeus monodon) during frozen storage

The colour of commercial cooked black tiger prawns (Penaeus monodon) is a key quality requirement to ensure product is not rejected in wholesale markets. The colour, due to the carotenoid astaxanthin, can be impacted by frozen storage, but changes in colour or astaxanthin profile, during frozen storage, have not been studied in detail. Subsequently in this study, the aims were to define the astaxanthin (as cis, trans, mono-ester and di-ester forms) content, together with the colour properties, in both pleopods (legs) and abdominal segments. Changes in astaxanthin content and colour properties were further determined during frozen storage (−20°C). Total astaxanthin content was seen to decrease in all samples over time, with the rate of degradation being significantly greater (P < 0.05) in pleopods than abdomen. In both pleopods and abdomen, rate of degradation of esterified forms was significantly greater (P < 0.05) than non-esterified forms. Hue angle (increase), a* value (decrease) and L value (increase) were all seen to significantly change (P < 0.05) during storage, with changes being more prevalent in the pleopods. The pleopods are the key indicator of astaxanthin and colour loss in cooked black tiger prawns and preservation strategies are required to preserve astaxanthin and colour during frozen storage.

Factors affecting fertility in frozen-thawed boar sperm

Frozen-thawed boar sperm holds the potential to have an impact on the future of the swine industry. Utilization of this technology could improve a swine producer’s ability to access top-tier genetics from around the world, to improve efficiency, profitability, and the quality of product to meet consumer demands. Effective application of frozen-thawed sperm can help reduce the potential risk associated with devastating economic loss due to the spread of disease. Frozen storage of boar sperm also provides a safeguard in the event of disease outbreaks, as genetic material from paternal lines can be preserved and banked for repopulation purposes. Historically these benefits have been masked by reduction in fertility measures such as litter size. The reduced fertility results from the damage sustained by the sperm cell during cryopreservation. However, increased understanding of this damage has lead to improved cryopreservation methods, ultimately increasing post-thaw viability and fertility. Enhancements in breeding technology have also resulted in a better understanding of the AI methods required to achieve acceptable farrowing rates and litter size. Fertility following AI with frozen-thawed sperm is approaching that of liquid stored sperm, and producers may soon reap the benefits of this technology. This thesis will outline the current swine industry, opportunities for utilizing frozen-thawed sperm, the main components of sperm, why they are susceptible to damage, and current freezing and breeding practices. Objective 1 was to develop a cryopreservation protocol for our lab that resulted in consistent post-thaw motility ( ≥ 40%) that would eventually be used by Illinois boar studs for domestic and international sale of frozen sperm. Evaluation with both manual microscopy and CASA methods were conducted to verify quality. A preliminary breeding trial was then conducted to test the fertility of sperm frozen with this method. There were 41 ejaculates from 23 boars used for freezing. Sperm were frozen at 1.4x109 sperm/mL, averaging 55.61.1% (meanSE) motility, following thaw. The samples assessed were not different (P>0.05) in motility when compared with manual or CASA systems, and results were most reliable at a 1:40 sperm dilution. In the preliminary breeding trial, gilts (n=14) were inseminated with either a single (n=10) or double (n=4) AI using 1, 2, or 4x109 motile, frozen-thawed sperm. Overall, the resulting pregnancy rates averaged 71.4% and numbers of normal fetuses per litter averaged 15.51.3 per litter. A feasibility study for freezing cost per ejaculate was estimated at $275/ejaculate or $11/dose of frozen-thawed semen at standard doses of 5x109 total frozen-thawed sperm. This cost estimate did not include genetic value, fixed equipment costs, depreciation, or variable lab space fees. Objective 2 focused on the proper methods for breeding with frozen-thawed boar sperm to achieve fertility. Our hypothesis was that increased numbers of inseminations and increased numbers of motile frozen-thawed sperm would improve pregnancy rate and litter size. Results showed acceptable fertility at high sperm numbers, but also the optimal method for insemination with the lowest dose tested. Gilts (n=111) responded to synchronization methods and were bred with 1, 2, or 4x109 motile frozen-thawed sperm from six boars using a single AI at 32 h, or a double AI, with the first AI at 24 and 32 h following estrus. Ultrasound was conducted at 12 h intervals to estimate the time of ovulation. On day 32 of gestation, overall pregnancy rate (73%) and number of normal fetuses per litter (10.80.5) across all treatments did not differ, and were not affected by number of motile sperm, or the interaction of number of motile sperm and number of inseminations. However, the number of inseminations tended to affect (P=0.14) the number of normal fetuses. Litter size increased with a double AI compared to single AI. Multiple inseminations helped to allow insemination to occur close to ovulation in response to variation in the time of ovulation. Both pregnancy rate and number of normal fetuses were greater when the time of the AI at 32 h occurred closer to the estimated time of ovulation (P<0.05). In addition, other factors such as presence of an abnormal ovary at day 30 decreased (P<0.001) pregnancy rate, while boar affected number of normal fetuses (P<0.01). Analysis of our data using a fertility index revealed doses of 2x109 motile sperm with multiple AI can achieve acceptable fertility with use of less sperm, when compared to AI using 4x109 motile sperm. The methods described here will investigate the potential for improved fertility when using frozen-thawed sperm, while accounting for variation in time of ovulation.

Fresh Frozen Plasma in Hereditary Angioedema Crisis: To Give or Not To Give?

Objectives: Fresh frozen plasma (FFP) has been used in angioedema crises, however there is a risk of aggravating the symptoms as well as transmitting infections. In this report, the authors emphasize the dangers of this therapy. Materials and methods: A 25-year-old woman with hereditary angioedema (HAE) was treated with FFP after which her symptoms escalated. Results: Administration of purified C1-inhibitor (C1-INH) resulted in relief of her symptoms. Conclusions: FFP is to be avoided in a HAE crisis. Newer therapies for angioedema are preferred.

Effect of grape seed extract on oxidative, color and sensory stability of pre-cooked, frozen, re-heated beef sausage model system

The processing of meats at the factory level can trigger the onset of lipid oxidation, which can lead to meat quality deterioration. Warmed over flavor is an off-flavor, which is associated with oxidative deterioration in meat. To avoid or delay the auto-oxidation process in meat products, synthetic and natural antioxidants have been successfully used. Grape (Vitis Vinifera) is of special interest due to its high content of phenolic compounds. Grape seed extract sold commercially as a dietary supplement, has the potential to reduce lipid oxidation and WOF in cooked ground beef when added at 1%. The objective of study 1 was to compare the antioxidant activity of natural antioxidants including grape seed extract and some herbs belonging to the Lamiaciae family: rosemary (Rosmarinus Officinalis), sage (Salvia Officinalis) and oregano (Origanum Vulgare) with commercial synthetic antioxidants like BHT, BHA, propyl gallate and ascorbic acid using the ORAC assay. All sample solutions were prepared to contain 1.8 gm sample/10 ml solvent. The highest antioxidant activity was observed for the grape seed extract sample (359.75 µM TE), while the lowest was observed for BHA, propyl gallate and rosemary also showed higher antioxidant potential with ORAC values above 300 μmol TE/g. ORAC values obtained for ascorbic acid and Sage were between 250-300μ mol TE/g while lowest values were obtained for Butylated Hydroxytoluene (28.50 µM TE). Based on the high ORAC values obtained for grape seed extract, we can conclude that byproducts of the wine/grape industry have antioxidant potential comparable to or better than those present in synthetic counterparts. The objective of study 2 was to compare three levels of grape seed extract (GSE) to commonly used antioxidants in a pre-cooked, frozen, stored beef and pork sausage model system. Antioxidants added for comparison with control included grape seed extract (100, 300, 500 ppm), ascorbic acid (AA, 100 ppm of fat) and propyl gallate (PG, 100 ppm of fat). Product was formed into rolls, frozen, sliced into patties, cooked on a flat griddle to 70C, overwrapped in PVC, and then frozen at –18C for 4 months. GSE- and PG-containing samples retained their fresh cooked beef odor and flavor longer (p<0.05) than controls during storage. Rancid odor and flavor scores of GSE-containing samples were lower (p<0.05) than those of controls after 4 months of storage. The L* value of all samples increased (p<0.05) during storage. Thiobarbituric acid reactive substances (TBARS) of the control and AA-containing samples increased (p<0.05); those of GSE-containing samples did not change significantly (p>0.05) over the storage period.

Odour flux from litter of broiler chickens fed diets differing in protein levels and additives

The effect of dietary crude protein (CP) and additives on odour flux from broiler litter was investigated using 180 day-old Ross 308 male chicks randomly allocated to five dietary treatments with three replications of 12 birds each. A 5×3 factorial arrangement of treatments was employed. Factors were: diet (low CP, high CP, high CP+antibiotic, high CP+probiotic, high CP+saponin) and age (15, 29, 35 days). Low CP (LCP) and high CP (HCP) diets differed in CP levels by 4.5-5%. The low CP diets were supplemented with L-valine, L-isoleucine, L-arginine, L-lysine, D,L-methionine and L-threonine. The antibiotic used was Zn Bacitracin, the probiotic was a blend of three Bacillus subtilis strains and the saponin came from a blend of Yucca and Quillaja. Odorants were measured from litter headspace using a flux hood and selective ion flow tube mass spectrometry (SIFT-MS). Results were log tranformed and analysed by two-way ANOVA with repeated measures using JMP statistical software v.8, and means were separated by Tukey's HSD test at P<0.05.The results showed that LCP group produced lower flux of dimethyl amine, trimethyl amine, H2S, NH3 and phenol in litter compared to HCP group (P<0.05). Similarly, HCP+probiotic group produced lower flux of H2S (P<0.05) and HCP+saponin group produced lower flux of trimethylamine and phenol in litter compared to HCP group (P<0.05). The dietary treatments tended (P=0.065) to have higher flux of methanethiol in HCP group compared to others. There was a diet x age interaction for litter flux of diacetyl, acetoin, 3-methyl-1-butanol, 3-methylbutanal, ethanethiol, propionic acid and hexane (P<0.05). Concentrations of diacetyl, acetoin, propionic acid and hexane in litter were higher from LCP group compared to all other treatments on d 35 (P<0.05) but not on days 15 and 29. Thus, the low CP diet, Bacillus subtilis based probiotic and Yucca/Quillaja based saponin were effective in reducing the emissions of some key odorants from broiler litter.

Effect of poultry viscera meal inclusion in broiler diets in different rearing periods on performance, carcass and parts yields

The objective of the present study was to evaluate the effect of the dietary inclusion of poultry viscera meal (VM) on broiler performance and carcass, parts, and abdominal fat yields in broilers by replacing a diet containing VM with a strictly vegetable diet and vice-versa. A number of 720 one-day-old broiler chicks were randomly distributed in 6 groups: G1-basal diet (BD) - corn and soybean based meal, with no VM from 1 to 42 days of age, G2- 8% VM diet from 1 to 42 days, G3- BD from 1 to 21 and 8% VM diet from 22 to 42 days, G4- BD from 1 to 35 and 8% VM diet from 36 to 42 days, G5- 8% VM diet from 1 to 21 days and BD from 22 to 42 days, G6- 8% VM diet from 1 to 35 and BD from 36 to 42 days. Average body weight, weight gain, feed intake, feed conversion ratio (FCR), production efficiency index, and mortality were determined from 1 to 42 days. There was no effect of treatments on performance or mortality, except for FCR, which was significantly better in the group fed VM from 1 to 35 days and withdrawn at the end of rearing (36-42 days). Also, there were no differences in carcass, parts, and abdominal fat yields, showing that VM in broiler diets does not influence yield parameters.