This Technical Research Communication describes a new method by which thermally treated Coxiella in milk products may be grown in a liquid growth medium and quantitated using an MPN-PCR assay. Coxiella is generally not used in studies on thermal and non-thermal processing of milk due to the need for specialized and highly laborious techniques such as animal assays and tissue culture for determining viability. Recently, a liquid growth medium (ACCM-2) and modified atmosphere were used to grow Coxiella from pure cultures, infected mouse tissues, and clinical samples, however, the ability to grow Coxiella from a food such as milk has not been shown. The potential ability to enrich Coxiella directly from contaminated milk presents a new avenue for conducting pasteurization research in which the viability of heat-treated or injured cultures could be more easily determined through direct enrichment of Coxiella in ACCM-2. ACCM-2 medium allowed enrichment of Coxiella from bovine whole milk and cream, whole goat, and whole camel milks but not whole water buffalo milk. Enrichment was possible from whole bovine milk containing as few as 6 Coxiella ge/ml of milk. The applicability of this ACCM-2 enrichment method was shown when using an MPN-PCR assay to quantitate the number of viable Coxiella remaining in whole bovine milk after 64 °C thermal treatment for up to 10 min.

This research communication aimed to evaluate the level of immunoglobulin E from lactic acid bacteria (LAB) that are used in dairy industries. Previous studies have demonstrated that workers report symptoms of irritation and are frequently IgG-sensitised to LAB. Workers (n = 44) from a probiotic production unity and the control lab were seen by a medical practitioner and responded to an occupational questionnaire. Specific IgE by the DELFIA® technique against 6 strains of LAB were measured on 44 exposed workers and 31 controls sera. Levels of specific IgE were low and no difference was observed between the two groups. This lack of IgE response could be explained by a healthy worker effect, an efficient implementation of personal protective equipment or by an absence of allergic mechanisms to account for the self-reported irritative symptoms. Despite the high concentrations of LAB, preventive measures are effective enough to guarantee no allergic effect and to prevent other adverse health effects. The implementation of preventive measures to avoid or reduce exposure to dust of LAB, and more generally to milk powder, is recommended in all dairy industry.

This research paper addresses the hypothesis that RagD is a key signalling factor that regulates amino acid (AA) mediated-casein synthesis and cell proliferation in cow mammary epithelial cells (CMECs). The expression of RagD was analysed at different times during pregnancy and lactation in bovine mammary tissue from dairy cows. We showed that expression of RagD at lactation period was higher (P < 0·05) than that at pregnancy period. When CMECs were treated with methionine (Met) or lysine (Lys), expression of RagD, β-casein (CSN2), mTOR and p-mTOR, and cell proliferation were increased. Further, when CMECs were treated to overexpress RagD, expression of CSN2, mTOR and p-mTOR, and cell proliferation were up-regulated. Furthermore, the increase in expression of CSN2, mTOR and p-mTOR, and cell proliferation in response to Met or Lys supply was inhibited by inhibiting RagD, and those effects were reversed in the overexpression model. When CMECs were treated with RagD overexpression together with mTOR inhibition or conversely with RagD inhibition together with mTOR overexpression, results showed that the increase in expression of CSN2 and cell proliferation in response to RagD overexpression was prevented by inhibiting mTOR, and those effects were reversed by overexpressing mTOR. The interaction of RagD with subunit proteins of mTORC1 was analysed, and the result showed that RagD interacted with Raptor. CMECs were treated with Raptor inhibition, and the result showed that the increase in expression of mTOR and p-mTOR in response to RagD overexpression was inhibited by inhibiting Raptor.

In conclusion, our study showed that RagD is an important activation factor of mTORC1 in CMECs, activating AA-mediated casein synthesis and cell proliferation, potentially acting via Raptor.

This Research Communication addresses the hypothesis that fat, protein, lactose and total solids content can be predicted using daily milk yield (DMY), pH, electrical conductivity (MEC) and refractive index (RI) of milk as predictors. It also addresses the possibility of these measurements being used for on-farm benchmarking activities towards selecting the highest yielding animals and flocks regarding milk quality traits (MQT). A total of 308 purebred Frizarta ewes were used for the study. From each individual ewe, a composite milk sample was collected. pH, MEC and RI of milk were measured and the samples were assayed for fat, protein, lactose and total solids content, using an automatic infrared milk analyser. The predictive value of DMY, pH, MEC and RI of milk on its MQT was assessed using multiple linear regression analysis. Significant regression equations were produced for all of the studied traits. RI and MEC were significant and reliable predictors for all studied MQT, whereas DMY was a significant predictor for most MQT with the exception of protein content. pH was a marginally significant predictor for some of the MQTs at the initial development of the equations but proved unreliable after bootstraping. Using these equations a number of ewes varying from 75 (for fat) to 97 (for protein) out of the 100 highest MQT yielders were correctly predicted, whereas, none of the ewes out of the 100 lowest MQT yielders was mispredicted as a high yielder for protein-, lactose- and total solids- content. Three out of 100 lowest fat-yielders were mispredicted as high fat-yielders. Similar equations can be used for benchmarking activities towards selecting the highest protein-, fat-, lactose- and total solids- yielding animals and flocks in cases where laboratories for MQT analyses are not readily available or the cost of chemical analyses is high. The method can be regarded as a handy tool for the dairy industry to readily assess milk quality at the farm level.

Scientific studies demonstrate the importance of intestinal microbiota to human health and how probiotic microorganisms can positively affect health when administered regularly and in adequate amounts. Probiotic bacteria can be part of fermented products and their functional importance is associated mainly with their metabolism. They are thought to benefit individuals to maintain their health and also to strengthen resistance against various types of diseases. The acceptance of probiotic cultures and products by consumers increased when these bacteria were marketed as natural cultures that help in digestion and health. Considering this, the food industry has an increasing demand for new candidates as probiotic cultures, and the dairy industry has a particular interest for fermented milks and other dairy products, since these are the most common food vehicles for probiotic cultures. Therefore, the dairy industries are increasingly seeking to improve their products with these beneficial bacteria. However, the legal peculiarities and excess of control agencies in Brazil makes the registration of these products and the collection of data very complex. Prospective analysis suggests that probiotic foods have the potential to effect a considerable expansion of the dairy industry, allowing the dairy sector to grow and for these products to be increasingly sought by consumers globally. For this, not only actions on research and innovation are necessary, but also official clarifications on the claims for considerations of microbiological security and functionality of these products. This review aims to elucidate important probiotic research regarding the isolation and characterization of beneficial cultures in Brazil, and to demonstrate the relevance of the dairy chain as a potential source of novel cultures for the development of new probiotic products to expand the Brazilian dairy industry.

Near infrared spectroscopy (NIRS) has the potential to estimate contents of fatty acids (FA) in milk frequently at-farm or during daily milking routine. In this study, a total of 738 raw milk spectra collected from 33 Holstein cows over a period of 30 weeks were recorded. Reference data on FA composition in milk and in milk fat were analysed in laboratory. Calibration models were calculated for single FA and groups of FA in milk and in milk fat. Validation resulted in sufficient Ratio of Prediction to Deviation (RPD) values for some single FA and in higher RPD values for groups of FA when concentrations of FA in milk were predicted. Since the concentrations of most FA in milk are highly correlated with milk fat content, the prediction of FA contents in milk fat is more meaningful when independent predictions are intended. The accuracy of predicting single FA concentrations in milk fat is rather poor for most FA but still comparable to alternative analysing methods such as MIR analysis. The estimation of different groups of FA in milk fat resulted in an improved accuracy based on higher RPD values, which was sufficient to mirror the development in the different lactation phases. The course of cow individual long chain fatty acid (LCFA) concentration in the early lactation stage can be an indicator for body fat mobilisation. The accurate estimation of the extent and duration of body fat mobilisation in cow individuals was rather difficult with NIR predicted LCFA concentrations and would require a higher measuring frequency than applied in this study.

This Research Communication investigated the potential of mid-infrared spectroscopy to predict detailed mineral composition of bovine milk. A total of 153 bulk milk samples were analysed for contents of Ca, Cl, Cu, Fe, K, Mg, Na, P and Zn. Also, soluble and colloidal fractions of Ca, Mg and P were quantified. For each milk sample the mid-infrared spectrum was captured and stored. Prediction models were developed using partial least squares regression and the accuracy of prediction was evaluated using both cross- and external validation. The proportion of variance explained by the prediction models in cross-validation ranged from 34% (Na) to 77% (total P), and it ranged from 13% (soluble Mg) to 54% (Cl−) in external validation. The ratio of the standard deviation of each trait to the standard error of prediction in external validation, which is an indicator of the practical utility of the prediction model, was low and never greater than 2. Results from the current study supported the limited usefulness of mid-infrared spectroscopy to predict minerals present in low concentration in bulk milk. For major mineral components, results from the present research did not match previous findings demonstrating the need for further studies using larger reference datasets.

The research reported here seeks to evaluate the allergenicity and antigenicity of different mammalian and plant-based milks/milk substitutes in healthy subjects. We used ELISA to measure IgE and IgG antibodies against cow, goat, sheep, camel, human milks, and soy, almond, and coconut plant-based milk substitutes, as well as IgA antibodies against all these apart from human milk, in 500 individuals in order to find the percentage of antibody elevation. IgG and IgE positivity showed that human milk was the least antigenic and allergenic, followed by camel milk. Cow's milk showed the highest percentage of elevation or reactivity. Among plant-based milk substitutes, the almond-based substitute was the most allergenic with the highest IgE reactivity, while the coconut milk substitute was lowest. For IgG and IgA immuno-reactivity, soy was first, with coconut again the lowest. We found IgE and IgG immune reactivity against coconut, almond and soymilks in some individuals who were non-reactive to mammalian milk, therefore, we should not assume that consumption of these milks is automatically without risk of allergenic response. We selected 24 samples out of the original 500 for the measurement of IgE antibodies against five different types of cow's milk, from non-organic to organic, A1 and A2. Statistical variance analysis detected no significant difference in IgE, IgG and IgA immune reactivities of the five different cow milks. Our results showed that if an individual is immuno-reactive to cow's milk, organic or not, the probability of reacting to goat and sheep milk is very high. Overall, the results presented here showed that for individuals allergic to cow's milk, the least allergenic alternatives in descending order are human, camel, sheep, and goat milks. Before choosing an alternative for cow's milk, one must go through accurate and quantitative blood testing for determination of IgE, IgG and IgA antibodies against different mammalian and plant-based milks/milk substitutes.

The aim of the studies reported in the Research Communication was to develop a rapid spectroscopic technique as an alternative method for the classification and discrimination of milk sources by Fourier transform infrared spectroscopy (FTIR). Cow, sheep and water buffalo milk samples were collected from various local milk producers in Istanbul, Turkey. In addition, various brands of packaged milk were purchased locally. Spectrums were obtained according to milk species origin and binary mixtures prepared in increments of 10% (10, 20, 30, 40, 50, 60, 70, 80 and 90%) for each sample analysed in FTIR spectroscopy. A successful milk species (cow, sheep, and water buffalo) discrimination and classification were achieved utilising Hierarchical cluster and principle component analyses (PCA) on the basis of Euclidean distance and Ward's algorithm. Amide-I (1700–1600/cm) and Amide-II (1565–1520/cm) spectral bands were used in the chemometric method. The results of the study indicated that adulteration of milk samples can be quantitatively detected by the FTIR technique in a short time with high accuracy. In conclusion, this method could be used as a new alternative technique for routine analysis in authenticity control of milk species origin.

This Review describes the objectives and methodology of the DairyWater project as it aims to aid the Irish dairy processing industry in achieving sustainability as it expands. With the abolition of European milk quotas in March 2015, the Republic of Ireland saw a surge in milk production. The DairyWater project was established in anticipation of this expansion of the Irish dairy sector in order to develop innovative solutions for the efficient management of water consumption, wastewater treatment and the resulting energy use within the country's dairy processing industry. Therefore, the project can be divided into three main thematic areas: dairy wastewater treatment technologies and microbial analysis, water re-use and rainwater harvesting and environmental assessment. In order to ensure the project remains as relevant as possible to the industry, a project advisory board containing key industry stakeholders has been established. To date, a number of large scale studies, using data obtained directly from the Irish dairy industry, have been performed. Additionally, pilot-scale wastewater treatment (intermittently aerated sequencing batch reactor) and tertiary treatment (flow-through pulsed ultraviolet system) technologies have been demonstrated within the project. Further details on selected aspects of the project are discussed in greater detail in the subsequent cluster of research communications.

We examined the latent structure of 26 cheese related phenotypes in dairy cattle. Traits related to milk yield and quality (8 traits), milk protein fractions (8 traits), coagulation and curd firmness indicators (CF, 5 traits) and cheese-making phenotypes (cheese yields (%CY) and nutrient recoveries in the curd (REC), 5 traits) were analysed through multivariate factor analysis (MFA) using a varimax rotation. All phenotypes were measured in 1264 Brown Swiss cows. Ten mutual orthogonal, latent variables (factors; Fs) were obtained explaining 74% of the original variability. These Fs captured basic concepts of the cheese-making process. More precisely, the first 4 Fs, sorted by variance explained, were able to capture the underlying structure of the CY percentage (F1: %CY), the CF process with time (F2: CFt), the milk and solids yield (F3: Yield) and the presence of nitrogen (N) in the cheese (F4: Cheese N). Moreover, 4 Fs (F5: as1-β-CN, F7: κ-β-CN, F8: as2-CN and F9: as1-CN-Ph) were related to the basic milk caseins and 1 factor was associated with the α-LA whey protein (F10: α-LA). A factor describing udder health status (F6: Udder health), mainly loaded on lactose, other nitrogen compounds in the milk and SCS, was also obtained. Further, we inferred the effects of some potential sources of variation (e.g. stage of lactation and parity) including feeding and management systems. Stage of lactation had a significant effect for 7 of the 10 Fs, followed by parity of the cow (3 Fs), dairy system and feeding (3 Fs). Our work demonstrates the usefulness of MFA in reducing a large number of variables to a few latent factors with biological meaning and representing groups of traits that describe a complex process like cheese-making. Such an approach would be a valuable tool for studying the influence of different production environments and individual animal factors on protein composition and cheese-making related traits.

The hypothesis of this research paper was that the physicochemical conditions in a low-pH, high-moisture white brined cheese such as Feta would make para-κ-casein vulnerable to residual chymosin activity during ripening and storage. It was important to address this hypothesis, since cheese para-κ-casein could theoretically be used for the assessment of the origin of cheese milk by means of various analytical methods. Feta cheese was manufactured from sheep milk and from four different mixtures of sheep and goat milk in triplicate. The para-κ-casein of Feta samples taken during 120 d of ripening and storage was estimated by means of cation-exchange HPLC and proteolysis was determined in terms of free amino groups. Despite their similarity, sheep and goat para-κ-casein were efficiently separated and the changes of their chromatographic areas indicated that hydrolysis took place during the first stage of ripening. In accordance to the evolution of free amino groups, para-κ-caseins remained stable thereafter. The hydrolysis pattern was not affected by the composition of the cheese milk mixture and after 120 d at least two thirds of the initial quantity remained intact. Considering the efficient separation of sheep and goat para-κ-caseins and their stability during Feta storage, the same method was used for the evaluation of the percentage of goat milk in the cheese milk. The actual and the estimated percentage of goat milk within the range 0–40 were strongly correlated (R = 0·997, n = 60) and the standard error of estimation was 0·914.

Cronobacter sakazakii is a life-threatening foodborne pathogen found in powdered infant formula and dairy products. Kefir is a dairy probiotic product and its antimicrobial activity against C. sakazakii was reported in our previous study. To identify key microorganisms that mediate growth suppression, we tested the antimicrobial activity of culture supernatants derived from lactic acid bacteria found in kefir. Lactobacillus kefiri DH5, L. kefiranofaciens DH101, and Bifidobacterium longum 720 (a commercial probiotic strain that served as a positive control) all significantly inhibited the growth of C. sakazakii ATCC 29544, delaying the initiation of exponential growth from 3 to 9 h in the nutrient broth. Among them, L. kefiri DH5 exerted the strongest antimicrobial effects against C. sakazakii, showing bactericidal effect at the addition of 300 µl of supernatant in 1 ml of nutrient broth. Interestingly, the supernatant of L. kefiri DH5 has higher pH and lower titrable acidity than that of L. kefiranofaciens DH101, suggesting metabolites produced by heterofermentation of L. kefiri acted more effectively to antagonise the growth of C. sakazakii. In addition, the supernatant of L. kefiri DH5 induced the leakage of cytoplasmic materials including nucleic acid and proteins, suggesting L. kefiri DH5 disrupted the cellular membrane integrity of C. sakazakii. Considering that pH neutralisation reduced the L. kefiri-dependent growth suppression, it is inferred that this activity is mainly due to organic acids produced during the fermentation process.

The studies described in the Research Communication aimed to describe the feasibility of using coal fly ash to synthesize nano-zeolite, and the ammonium and phosphate adsorption efficiencies of the nanomaterial in dairy wastewater. Chemical treatment of coal fly ash was conducted and changes observed. Samples treated with NaOH had an increased cation exchange capacity and P sorption index compared to the initial fly ash, due to particle modification from smooth surface to plate- and rod-shape crystals, referred to as nano-zeolite. Batch experiments were conducted by mixing coal fly ash and nano-zeolite with synthesized wastewater to study the effect of sorption time, pH values and dosage of nano-zeolite on ammonium and phosphate removal efficiency. The adsorption process reached equilibrium in a very short time (less than 60 min), which suggests a potential for fast immobilization of pollutants. The concentration of ammonium decreased from 118 to 35 mg/l (71% removal) while the concentration of phosphate decreased from 52 to 45 mg/l. The removal efficiency of ammonium was 36·6, 51·8 and 70·9% at pH 3, 7 and 10, respectively whilst that of phosphate increased dramatically with decreased slurry pH (92·1, 47·3 and 12·3% at pH 3, 7 and 10, respectively). Nano-zeolite could be a potential absorbent for fast immobilization of ammonium but not phosphate. Surface modification of nano-zeolite could be introduced in order to enhance the pollutants removal efficiency.

This Research Communication describes an investigation into the viability of an Intermittently Aerated Sequencing Batch Reactor (IASBR) for the treatment of dairy processing wastewater at laboratory-scale. A number of operational parameters have been varied and the effect has been monitored in order to determine optimal conditions for maximising removal efficiencies. These operational parameters include Hydraulic Retention Time (HRT), Solids Retention Time (SRT), aeration rate and cycle length. Real dairy processing wastewater and synthetic wastewater have been treated using three laboratory-scale IASBR units in a temperature controlled room. When the operational conditions were established, the units were seeded using sludge from a municipal wastewater treatment plant for the first experiment, and sludge from a dairy processing factory for the second and third experiment. In experiment three, the reactors were fed on real wastewater from the wastewater treatment plant at this dairy processing factory. These laboratory-scale systems will be used to demonstrate over time that the IASBR system is a consistent, viable option for treatment of dairy processing wastewater in this sector. In this study, the capacity of a biological system to remove both nitrogen and phosphorus within one reactor will be demonstrated. The initial operational parameters for a pilot-scale IASBR system will be derived from the results of the study.

The present study focused on the utilisation of High Intensity Light Pulses (HILP) treatment to preserve mozzarella cheese. First, the susceptibility of Pseudomonas fluorescens and Enterobacteriaceae to HILP (fluences from 0·39 to 28·0 J/cm2) in a transparent liquid was evaluated (in-vitro tests). Afterwards, the effects on inoculated mozzarella cheese were also assessed. Then untreated (Control) and HILP treated samples were packaged and stored at 10 °C for 2 weeks. Enterobacteriaceae, Pseudomonas spp. and pH were monitored during storage. In a transparent liquid (in-vitro tests) there was a significant microbial inactivation just with 2 s of treatment. On the inoculated cheese a relevant microbial reduction of about 1 log cycle was observed, according to the exposure to the treatments. For Pseudomonas spp. in particular, in the treated samples, the microbiological acceptability limit (106 cfu/g) was never reached after 2 weeks of refrigerated storage. To sum up, the efficacy of this treatment is very interesting because a microbial reduction was observed in treated samples. HILP treatment is able to control the microbial growth and may be considered a promising way to decontaminate the surface of mozzarella cheese.

In this study, the relationship between the dissociation of κ-casein from casein micelles due to heat-induced denaturation and the strength of acid milk gel was investigated. The κ-casein-dissociated micelles were fractionated by gel filtration chromatography and two-dimensional polyacrylamide gel electrophoresis, and their zeta potential and surface hydrophobicity were measured. The negative charge of the κ-casein-dissociated micelles was lower than that of native micelles, and micellar surface hydrophobicity was higher. For confirmation, the isoelectric point of the casein micelles was measured. The κ-casein-dissociated micelles were found to cohere at an earlier stage of acidification than the native micelles. These results demonstrated that the heat-induced increase in the strength of acid milk gel was partly due to the decrease in micellar surface charge and partly to the increase in surface hydrophobicity caused by the dissociation of κ-casein.

This Research Communication describes the initial operation of a pilot-scale intermittently aerated sequencing batch reactor system, which is located at an Irish dairy processing factory. Laboratory-scale research has facilitated the design specifications and operational parameters necessary for the construction and running of a pilot-scale. Laboratory scale research was necessary prior to the pilot scale system to ensure high quality treatment and nutrient removal efficiencies. The pilot system operates with a hydraulic retention time of 4 d, a solids retention time of 16 d and a cycle length of 12 hours. There are 4 non-aeration and aeration phases within the system's react phase. This system has a 3000 l working volume, treating 375 l of wastewater per cycle, 750 l daily. The system was seeded from an aeration tank at the dairy processing factory where the unit is located. The system is operating with the goal to remove both nitrogen and phosphorus from the wastewater biologically, reducing the need for chemical treatment. Currently, the system is performing with high efficiency, treating the wastewater to an acceptable level according to the Irish Environmental Protection Agency for discharge into surrounding water bodies. Therefore, the initial removal results demonstrate this technology's suitability for the treatment of high strength dairy wastewaters.