Evaluation of Brix Refractometry to Estimate Immunoglobulin G Content in Buffalo Colostrum and Neonatal Calf Serum.

Brix refractometry has been widely demonstrated to be a useful tool for monitoring colostrum management program and passive immunity transfer (PIT) in Bovines, but its suitability has never been verified in Buffalo. Therefore, the objective of this study was to evaluate the utility of a simple and rapid tool such as a digital Brix refractometer to estimate colostrum quality and for evaluating the success of passive transfer of immunoglobulin G (IgG) in Buffalo calves. The optimal cut points levels for Brix Refractometry for distinguishing good- and poor-quality colostrum and for assessing the adequacy of passive immunity transfer in calves were determined. For this aim, 26 first-milking maternal colostrum (MC) were collected from first-calf heifers. Blood samples were obtained from their calves at birth (T0) and 72 hours after (T3). Colostrum and Serum IgG content were determined by indirect enzyme-linked immunosorbent assay (ELISA), whereas total protein (TP, g/dL) and percentage Brix (%Brix) by means of a digital Brix refractometer. The mean colostrum IgG was 64.9 ± 29.3 mg/mL. The mean serum %Brix at T3 was 9.6 ± 0.9 %. The mean serum IgG content at T3 was 11.1 ± 2.0 mg/mL. Pearson's correlation coefficient (rp) was determined between Brix and ELISA measurements: colostrum %Brix showed a significant correlation with serum %Brix (rp = 0.82, p < 0.001); serum %Brix was highly correlated with serum TP (STP, g/dL) (rp = 0.98, p < 0.001) and serum IgG (mg/mL) (rp = 0.85, p < 0.001). A cut point of 18% Brix to estimate samples of MC ≥ 50 mg/mL from first-calf heifers was more appropriate for the buffalo. A cut point of 8.4% Brix resulted in the greatest percentage of calf serum samples being correctly classified. Based on our findings, a digital Brix refractometer could be a useful tool to monitor colostrum quality and to estimate PIT in Buffalo calves.

Oxidative Status of Marchigiana Beef Enriched in n-3 Fatty Acids and Vitamin E, Treated With a Blend of Oregano and Rosemary Essential Oils.

The aim of this study was to evaluate the effects of basal dietary supplementation with linseed and linseed plus vitamin E in Marchigiana young bulls on the instrumental color, oxidative stability, and fatty acid composition of beef steaks with and without treatment with a blend of essential oils (oregano and rosemary) after storage times of 0, 3, 6, and 9 days. The fatty acid composition was evaluated in meat after 0 and 9 days of storage. No variation in the fatty acid composition of each type of meat was observed after storage. The use of dietary linseed and vitamin E, compared with linseed alone, significantly improved the oxidative stability, lightness (L*), and redness (a*) of the meat. A higher degree of oxidation was observed in meat samples from animals in the group fed linseed (L) followed by the control group (C) and the group fed linseed and vitamin E (LE). Moreover, the essential oils treatment (O) exerts a significant effect on beef oxidative stability of the group LE more than groups C and L during storage. The fatty acid composition of the meat was also affected by the diet: levels of healthful fatty acids (PUFA, n-3 PUFA, and CLA) were higher in meat from the groups that received linseed than the control group. Our results suggest that dietary vitamin E and treatment with essential oils are effective approaches to preserving the stability of beef cattle receiving a diet enriched in unsaturated fatty acids for up to 9 days of storage.

Effects of three different diets on the fatty acid profile and sensory properties of fresh Pecorino cheese "Primo Sale".

OBJECTIVE: This study aimed to evaluate and compare the effects of three different diets on the fatty acids (FA) profile and sensory properties of a characteristic Italian fresh cheese: Pecorino "Primo Sale" (PS). METHODS: Fifty-four sheep were divided into three feeding groups: total mixed ration (TMR) enriched with extruded linseed (TL), control diet with TMR without any integration (TC), and pasture (P). During cheese production, six cheeses per experimental group were produced each week, stored for 10 days at 4°C, and then analyzed for chemical composition, FA profile, and sensory properties. RESULTS: Saturated fatty acids (SFA) were significantly higher in PS from group TC (82.11%) than in cheese from other two groups (P 75.48% and TL 66.83%). TL and P groups presented higher values of polyunsaturated fatty acids, 4.35 and 3.65%, respectively, than that of TC group (2.31%). The lowest SFA/unsaturated fatty acid ratio was found in TL and P groups, while the highest was found in the TC group. Vaccenic acid and conjugated linoleic acid (CLA) were higher in group P (p<0.05) than in groups L and TC. Sensory properties of cheese from group P received the highest scores for odor intensity and friability, while control group had a greater chewing consistency. Overall, all cheeses received good scores for acceptability. CONCLUSION: In conclusion, this study showed how the integration of extruded linseed improved the FA profile of fresh pecorino cheese PS preserving its sensory properties. Levels of CLA in the PS group achieved using this approach was not higher than that in a grazing diet. Cheeses from groups P and L contained a higher value of FA, with nutritional implications for humans, when compared with un-supplemented diet. Linseed may be a good feeding strategy when pasture is not available.

Influence of Iodine Feeding on Microbiological and Physico-Chemical Characteristics and Biogenic Amines Content in a Raw Ewes' Milk Cheese.

Iodine is an essential trace element involved in the regulation of thyroid metabolism and antioxidant status in humans and animals. The aim of this study was to evaluate the effect of ewes’ dietary iodine supplementation on biogenic amines content as well as microbiological and physico-chemical characteristics in a raw milk cheese at different ripening times (milk, curd, and 2, 7, 15, 30, 60, and 90 days). Two cheese-making trials were carried out using milk from ewes fed with unifeed (Cheese A) or with the same concentrate enriched with iodine (Cheese B). The results indicated that the counts of principal microbial groups and physico-chemical characteristics were quite similar in both cheeses at day 90. Cheese B was characterized by a higher content of biogenic amines and propionic acid. Propionic bacteria were found in both cheeses mainly in Trial B in agreement with the higher content of propionic acid detected.

Influence of pig rennet on proteolysis, organic acids content and microbiota of Pecorino di Farindola, a traditional Italian ewe's raw milk cheese.

The use of pig rennet is very ancient and in Italy is only applied in the manufacture of Pecorino di Farindola cheese. In order to evaluate the key role of this rennet in the establishment of peculiar features of Pecorino di Farindola, cheeses made from raw ewes' milk using calf (A) and kid (B) rennets were compared to those produced with pig (C) rennet. The use of pig rennet for Pecorino di Farindola cheese making confers physico-chemical and proteolytic characteristics that differentiate it from cheeses produced with other coagulants. However, no microbiological differences were observed. Chesses made with pig and kid rennets were characterised by higher proteolysis after 7 days of ripening. The content of isovaleric and propionic acids at the end of ripening was correlated with the presence of propionibacteria.

Phenolic compounds in grains, sprouts and wheatgrass of hulled and non-hulled wheat species.

BACKGROUND: The use of sprouts and young plantlets in human nutrition is increasing because they often contain phytochemicals and other high value nutrients. This is also the case for wheat, although there is no literature for hulled wheat species. Thus we determined total polyphenols, phenolic acids (PAs), fibre and minerals in grains, 5-day-old sprouts and 12-day-old wheatgrass of einkorn (cv. Monlis), emmer (cvs Augeo, Rosso Rubino, Zefiro), spelt (cvs Pietro, Giuseppe), durum wheat (cv. Creso) and soft wheat (cv. Orso). RESULTS: Grains of einkorn and emmer contained twice bound PAs as compared to soft and durum wheat and spelt, with p-coumaric acid accounting for about 50% of total bound PAs. In wheatgrass, differences between species for bound PAs decreased due to a decrease in einkorn and emmer and an increase in soft and durum wheat. In all species, total phenols and free PAs increased passing from grains to sprouts and wheatgrass. Neutral and acid detergent fibre content increased with sprouting only in einkorn and emmer. CONCLUSION: Our evidence suggests that the grains of einkorn and emmer and the sprouts and wheatgrass of all Triticum species might potentially be valuable for the development of functional foods.

High content of biogenic amines in Pecorino cheeses.

Pecorino refers to Italian cheeses made exclusively from raw or pasteurized ewes' milk, characterized by a high content of fat matter and it is mainly produced in the Middle and South of Italy by traditional procedures. The autochthonous microbiota plays an important role in the organoleptic traits of Pecorino cheese and it can influence biogenic amines (BA) content. The aim of this study was to characterize from microbiological and chemical point of view 12 randomly purchased commercial cheeses produced in Abruzzo region. Moreover, the BA content and the bacteria showing a decarboxylating activity were detected. For this purpose, a real-time quantitative PCR (qPCR) was applied to evaluate histamine and tyramine-producers. The samples were well differentiated for microbial groups composition, such as aerobic mesophilic bacteria, Enterobacteriaceae, coagulase-negative staphylococci, yeasts, enterococci, mesophilic and thermophilic lactobacilli. Pathogens such as Salmonella spp., Listeria monocytogenes and Escherichia coli O157:H7 were absent in all samples. In most samples the content of BA resulted to be high, with prevalence of histamine and tyramine. In particular, total BA content reached 5861 mg/kg in Pecorino di Fossa cheese. The qPCR method resulted to be very useful to understand the role of autochthonous Pecorino cheese microbiota on BA accumulation in many different products. In fact, since the ability of microorganisms to decarboxylate aminoacids is highly variable being in most cases strain-specific, the detection of bacteria possessing this activity is important to estimate the risk of BA cheese content.

High-performance liquid chromatography with post-column derivatisation and fluorescence detection for sensitive determination of aflatoxin M1 in milk and cheese.

A new HPLC method with fluorescence detection using pyridinium hydrobromide perbromide as a post-column derivatising agent has been developed to determine aflatoxin M1 in milk and cheese. The detection limits were 1 ng/kg for milk and 5 ng/kg for cheese. The calibration curve was linear from 0.001 to 0.1 ng injected. The method includes a preliminary C18-SPE clean-up and the average recoveries of Aflatoxin M1 from milk and cheese, spiked at levels of 25-75 ng/kg and 100-300 ng/kg, respectively, were 90 and 76%; the precision (RSDr) ranged from 1.7 to 2.6% for milk and from 3.5 to 6.5% for cheese. The method is rapid, easily automatable and therefore useful for accurate and precise screening of aflatoxin M1 in milk and cheese.

A rapid and sensitive 384-well microtitre format chemiluminescent enzyme immunoassay for 19-nortestosterone.

We developed a competitive chemiluminescent (CL) enzyme immunoassay for rapid, sensitive analysis of 19-nortestosterone (19-NT) in bovine urine. Anti-19-NT polyclonal antibodies were raised in rabbits using a 19-NT-hemisuccinate derivative conjugated with ovalbumin; the derivative was also conjugated with horseradish peroxidase (HRP) as a label. Antibodies were immobilized on 384-well black polystyrene microtitre plates and HRP-labelled 19-NT activity was measured using an efficient chemiluminescent substrate (SuperSignal ELISA Femto) after 3 min incubation. Emitted light was recorded using a conventional, photomultiplier-tube-based microtitre plate reader or a sensitive back-illuminated, cooled CCD camera. The developed method fulfils all the requirements of precision (intra- and inter-assay CV < 10%) and accuracy (mean recovery 94-112%), with a detection limit of 0.03 ppb (1.1 x 10(-9) mol/L) in a urine matrix. Chemiluminescence enhances detectability of the HRP-labelled tracer (thus lowering the limit of detection with respect to colorimetry) and reduces analysis time. The 384-well microtitre plate cuts the sample/reagent volume (20 microL), a five-fold reduction with respect to the conventional 96-well microtitre plate. The developed method is suitable for high-throughput screening of 19-NT in urine samples, with reduced costs as compared with conventional colorimetric enzyme immunoassays.