Simultaneous detection of mastitis pathogens, Staphylococcus aureus, Streptococcus uberis, and Streptococcus agalactiae by multiplex real-time polymerase chain reaction

The objective of this study was to develop a multiplex real-time polymerase chain reaction (PCR) method for simultaneous detection of Staphylococcus aureus, Streptococcus agalactiae, and Streptococcus uberis directly from milk. A genetic marker specific for Staph. aureus was used for primers and dual-labeled probe design. The target for Strep. agalactiae primers and dual-labeled probe was selected from the cfb gene encoding the Christie-Atkins-Munch-Petersen factor. The plasminogen activator gene was the target for primers and dual-labeled probe design for Strep. uberis. Quarter milk samples (n = 192) were analyzed by the multiplex real-time PCR assay and conventional microbiological methods. An additional 57 quarter milk samples were analyzed in a separate real-time PCR assay for Strep. agalactiae only. Using an overnight enrichment step, the real-time PCR technique correctly identified 96.4% of all quarter milk samples; 91.7% of Staph. aureus, 98.2% of Strep. agalactiae, and 100% of Strep. uberis. Results of conventional microbiological methods were used to determine the sensitivity and specificity of the multiplex real-time PCR procedure. The sensitivity of the procedure to correctly identify Staph. aureus, Strep. agalactiae, and Strep. uberis directly from milk was 95.5%, and the specificity was 99.6%. Results of this study indicate that the multiplex real-time PCR procedure has the potential to be a valuable diagnostic technique for simultaneous identification of Staph. aureus, Strep. agalactiae, and Strep. uberis directly from quarter milk samples.     

Effect of housing system, slaughter weight and slaughter strategy on carcass and meat quality, sex organ development and androstenone and skatole levels in Duroc finished entire male pigs

This study aimed at evaluating the effect of housing system (HS), slaughter weight (SW) and strategy (SS) on carcass a nd meat quality, sexual organ development and boar taint in entire males. Twelve pens of 10 pigs were used (two trials). Half of male pens were allowed visual contact with females (MF) and half with males (MM). Half MM or MF were slaughtered at 105 or 130 kg in trial 1, or penwise or by split marketing in trial 2 at 120 kg. Housing system showed no significant effect on carcass or meat quality. MF presented significantly longer testicles and heavier bulbourethral glands compared to MM. The distribution of androstenone and skatole levels was affected by SW but not by HS or SS, samples with androstenone >1 μg/g of the different groups falling within the range of 16 to 22%. All correlations between androstenone and sex organs were significant. Housing system and slaughter strategy did not reduce the risk of boar tainted carcasses.     

Insight into the glycation of milk proteins: an ESI- and MALDI-MS perspective (review)

Protein-reducing sugar conjugates are formed by the naturally occurring Maillard reaction, otherwise known as glycation. The Maillard reaction products (MRP) formed can provide novel and/or improved functionality compared to the unmodified protein. Understanding the chemistry of the Maillard reaction, the physicochemical properties of its products, and, more importantly, the inter-relationships between these properties and the specific functionality of a given MRP will help to define the potential of MRP as food ingredients in their own right. Recently, electrospray ionization-mass spectrometry (ESI-MS) and matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS) have acquired a leading role in the structural characterization of proteins. The ability of these techniques to provide detail about the nature and extent of protein modifications at a molecular level as well as conformational information provides new insight into the glycation process. This article reviews the role that ESI- and MALDI-MS have played in advancing our understanding of the glycation of milk proteins.     

Carcass and meat quality characteristics and fatty acid composition of tissues from Pietrain-crossed barrows and gilts fed an elevated monounsaturated fat diet

Fifty-one (Landrace∗Large White)∗Pietrain barrows and gilts were used to compare the effect of a diet rich in oleic acid (HO) by feeding a by-product of the olive industry (Greedy-Grass OLIVA®: 1.4% growing, 3.8% finishing) or a grain and soy diet (CONTROL) on carcass characteristics, meat quality and fatty acid profile of intramuscular and subcutaneous fat. Gilts had leaner (P < 0.05) carcasses with lower fat percentage in major primal cuts, and less (P < 0.05) saturated fat compared with barrows with no interaction (P > 0.05) between dietary treatment and gender. Source of dietary fat had no effect (P > 0.05) on primal cut yields, composition of major primal cuts, or carcass and meat quality characteristics. Intramuscular fat from HO fed pigs had higher (P < 0.05) percentage of saturated (SFA) and monounsaturated (MUFA) fatty acids, and lower (P < 0.05) polyunsaturated fatty acids (PUFA) and n-6:n-3 ratio compared with CONTROL animals (37.4% vs. 36.8%, 44.7% vs. 40.1%, 17.9% vs. 23.2%, and 18.9 vs. 21.8 ratio, respectively). Subcutaneous fat from pigs fed HO had greater (P < 0.05) MUFA percentage, lower (P < 0.05) SFA and PUFA percentage, and lower (P < 0.05) n-6:n-3 ratio than pigs fed CONTROL diet (51.4% vs. 48.0%, 30.5% vs. 32.9%, 18.1% vs. 20.1%, and 9.83 vs. 11.3 ratio, respectively). Intramuscular fat had higher proportion of SFA and lower of MUFA showing a higher degree of tissue saturation compared with subcutaneous fat. Feeding Greedy-Grass increased MUFA and decreased PUFA proportions in fat depots reducing the risk of production of carcasses that are soft and oily which result in lower technological and processing quality.     

Characterization of the lactic acid bacteria in ewe's milk and cheese from northwest Argentina

Indigenous lactic acid bacteria in ewe's milk and artisanal cheese were studied in four samples of fresh raw milk and four 1-month-old cheeses from the provinces of northwest Argentina. Mean growth counts on M17, MRS, and MSE agar media did not show significant differences (P < 0.05) in raw milk and cheeses. Isolates of lactic acid bacteria from milk were identified as Enterococcus (48%), lactococci (14%), leuconostocs (8%), and lactobacilli (30%). All lactococci were identified as Lactococcus lactis (subsp. lactis and subsp. cremoris). Lactobacilli were identified as Lactobacillus plantarum (92%) and Lactobacillus acidophilus (8%). Enterococci (59%) and lactobacilli (41%) were isolated from cheeses. L. plantarum (93%), L. acidophilus (5%), and Lactobacillus casei (2%) were most frequently isolated. L. lactis subsp. lactis biovar diacetylactis strains were considered as fast acid producers. L. lactis subsp. cremoris strains were slow acid producers. L. plantarum and L. casei strains identified from the cheeses showed slow acid production. The majority of the lactobacilli and Lactococcus lactis strains utilized citrate and produced diacetyl and acetoin in milk. Enzyme activities (API-ZYM tests) of lactococci were low, but activities of L. plantarum strains were considerably higher. The predominance of L. plantarum in artisanal cheese is probably important in the ripening of these cheeses due to their physiological and biochemical characteristics.     

Evaluation of two fluorogenic assays for identification of Streptococcus species isolated from bovine mammary glands

Liquid and agar assays that utilized 4-methylumbelliferyl-conjugated beta-D-glucuronide, beta-D-galactoside, or N-acetyl-beta-D-glucosaminide, and agglutination by Dolichos biflorus lectin were evaluated for identification of Streptococcus species isolated from bovine mammary glands. A greater number of Streptococcus uberis isolates were negative for N-acetyl-beta-D-glucosaminidase by the liquid assay compared with the agar assay. Enzyme profiles for Streptococcus dysgalactiae were similar by both assays. Streptococcus dysgalactiae was the only species that agglutinated when mixed with lectin from D. biflorus. Most Streptococcus agalactiae isolates were positive for beta-D-glucuronidase and beta-D-galactosidase by both assays. Two Streptococcus equinus strains had negative enzyme profiles by the liquid assay; however, both strains had enzyme profiles consistent for S. equinus by the agar method. Incorporation of 4-methylumbelliferyl-conjugated substrates into trypticase soy agar did not appear to alter agar characteristics and eliminated aliquoting substrates and inoculating tubes. More than one enzyme profile was produced per Streptococcus species or serogroup by both methods. However, some profiles were similar between species, which hindered accurate identification of Streptococcus species.     

木聚糖酶固态罐发酵及尾气在线测定

建立了可对 4个各自独立的 2L玻璃发酵罐同时通气培养和尾气检测的固态发酵系统(SSF) ,以 1 0 0g麸皮和 1 0 0g木屑及 1 5 0mLMandels营养盐溶液为培养基进行木霉T6培养 ,在实验条件范围内找出了最佳产木聚糖酶的环境条件 ,即温度为 3 0℃ ,pH6.0 ,接种量 2 5mL/罐 ( 2× 1 0 8～ 4×1 0 8个孢子 /mL) ,通过对木聚糖酶活力和尾气中CO2 变化的连续测定显示 ,菌体CO2 的释放与木聚糖酶合成具有一定的相关性 ,木聚糖酶的合成与尾气中CO2 含量之间存在着滞后关系 ,尾气CO2浓度在发酵 2 6h达到最高值 ,木聚糖酶活力在 3 9h才达峰值。通过对SSF尾气中CO2 的在线测定 ,能够初步了解SSF中菌丝体生长和产物合成之间的动力学关系     

Characterisation of young bulls of the Bruna dels Pirineus cattle breed (selected from old Brown Swiss) in relation to carcass, meat quality and biochemical traits

Seventy-four young bulls of the Bruna dels Pirineus beef cattle breed were reared in a typical production system and slaughtered at an average age of 381 days and live weight of 541 kg. The animals were evaluated for productive traits, carcass quality, meat quality, eating quality and biochemical characteristics of m. longissimus thoracis (LT). Biochemical measurements included intramuscular fat (IMF) and collagen proportion, haem pigment concentration, lactate dehydrogenase and isocitrate dehydrogenase activities and type I (MHC I) fibre percentage determined by ELISA. Bruna dels Pirineus bulls achieved high growth rates during the fattening period (1.63 kg/day) and showed good carcass quality, with a high dressing-out proportion (607 g/kg, hot carcass), a good conformation score (U; EUROP) and a moderate fatness score (3;1-5). Carcass composition was estimated from the sixth rib joint dissection (682 g/kg lean proportion, 127 g/kg total dissectable fat and 163 g/kg bone). IMF (24.3 g/kg) and MHC I (27.9%) showed high variabilities (CV>30%). Sensory analysis of LT included beef and livery odour and flavour intensity, and overall tenderness and juiciness assessment of loin samples (14-day ageing). Beef odour and flavour were slightly positively correlated with IMF and carcass fatness score (P<0.05). Fatness, MHC I, insoluble collagen and cooking losses tended to affect the livery flavour intensity positively. This variable was significantly higher in meat from bulls of lower carcass quality (i.e. lower conformation score, lower lean proportion; P<0.05) and higher type I fibre percentage. Loin overall tenderness and juiciness were not affected by the biochemical traits studied, however, they were negatively affected by cooking loss (P<0.05).     

A new method to determine the ratio of electricity production from fossil and biogenic sources in waste-to-Energy plants

New directives of the European Union require operators of waste-to-energy (WTE) plants to report the amount of electricity that is produced from renewable sources in the waste feed. Until now, the standard method to determine the portion of renewable electricity is sorting the wastes into defined fractions of fossil organic and biogenic waste components and determining the lower heating value of these fractions. Out of it the amount of electricity production from renewables is calculated. This practice is labor and cost intensive. Therefore, it is usually carried out once a year which provides only a snapshot analysis of limited significance. This paper proposes a method to calculate the portion of electricity produced from renewable materials in waste continuously by solving a set of equations. All data required are either available from literature or from operating data routinely measured in WTE plants. The advantages are statistically derived uncertainty of the result, temporal resolution of the result down to daily mean values, low implementation efforts, and virtually no operational costs. An example of the implementation of the method to a 60 000 tons per year WTE plant is given.