This research communication was designed to evaluate the effects of different levels of diet restriction on the composition and ethanol stability (MES) of raw bovine milk. This research was carried out using three electronic databases: Scopus, Pubmed and Web of Science. The main inclusion criteria were: (i) original research, (ii) use of alcohol (ethanol) test as a method to assess milk stability, (iii) measure different levels of feed restriction and (iv) allow access to the raw data of articles. Of the nine publications that addressed the subject filtered by the systematic review, seven fitted the selection criteria and were selected to perform the meta-analysis. Feed restriction (reduction of 20, 30, 40 and 50% of the dietary dry matter offered) decreased (P < 0.01) milk yield (−18%), ethanol stability (−5%), acidity (−4%), protein (−3%) and lactose (−2%) concentrations, but did not affect the values of pH, density, fat and total solids concentrations, nor somatic cell count. The correlation between milk yield and MES was low but positive and numerically higher in the control group compared with the restriction group. The milk of cows fed the control diet presented greater ethanol stability (76.5%) compared with milk of cows fed the restrictive diet (72.8%). This decrease by up to 4 percentage units due to restriction levels ranging from 20 to 50% of diet intake may cause limitations in milk processing at the dairy industry, increasing milk rejection.

The objectives of this research were (1) to study different factors affecting milk total bacterial count (TBC) and (2) to estimate the economic value associated with TBC in Holstein dairy herds in Iran. The relationships between bulk tank TBC and farm management and economic factors were examined on 56 randomly selected intensive dairy farms. Herd management factors associated with bulk tank TBC were determined using mixed linear models. The median bulk tank TBC for the sample herds was 299 (range 81–1185) × 103 cfu/ml. The average economic premium opportunity from bulk tank TBC was US$ 1.32 per ton of milk ranging from US$ 0.02 per ton of milk for herds applying wet tissue procedures as teat cleaning material and washing the water troughs three times per day to US$ 5.20 per ton of milk for herds with dirty barns. Results showed that the following management factors were associated with low TBC and high economic value: frequency of cleaning water troughs, teat cleaning material, the frequency of milk delivery to the processor, bedding material, herd size, education level of workers, udder washing material, material of milking parlor wall, frequency of disinfection of the calving area, presence of veterinarian, water quality control, having a hospital pen and barn hygiene. In conclusion, our findings highlight the need to pay more attention to farm management issues, particularly farm hygiene practices to reduce milk TBC and so reduce the economic burden of TBC in dairy herds in Iran.

We evaluated the possibility of increasing the storage temperature of raw milk for Provolone Valpadana cheesemaking, to identify the most suitable conditions of time and temperature for a pre-maturation process. We used Principal Component Analysis (PCA) to analyze the overall effects of different storage conditions on chemical, nutritional and technological characteristics of the raw milk. Four different thermal storage cycles, two at fixed temperature/time (6 and 12°C for 60 h) and two with two-phase thermal cycle (10 and 12°C for 15 h, followed by refrigeration at 4°C for 45 h) were studied. Although a moderate heterogeneity among raw milks from the 11 producers of Provolone Valpadana cheese was observed, PCA revealed the critical aspects of the extreme storage conditions (60 h of refrigeration). Some samples resulted in anomalous behaviors, probably related to unexpected fermentation phenomena occurring with increasing storage temperature. The acidification and the increase in the contents of lactic acid, soluble calcium, and degree of retinol isomerization observed in the anomalous samples can compromise the technological functionality of milk. Conversely, the storage with a two-phase thermal cycle did not lead to variations in any measured characteristic, suggesting that mild refrigeration conditions (10 or 12°C for 15 h followed by 4°C for 45 h) could be a good compromise in favoring milk pre-maturation without altering its quality characteristics.

Dairy goat farming is an important sector of the agricultural industry in Greece, with an annual total milk production exceeding 450 000 l and accounting for over 25% of all goat milk produced in the European Union; this milk is used mainly for cheese production. Despite the importance of goat milk for the agricultural sector in Greece, no systematic countrywide investigations in the bulk-tank milk of goats in Greece have been reported. Objectives were to investigate somatic cell counts (SCC) and total bacterial counts (TBC) in raw bulk-tank milk of goat herds in Greece, study factors influencing SCC and TBC therein and evaluate their possible associations with milk content. Throughout Greece, 119 dairy goat herds were visited for milk sampling for somatic cell counting, microbiological examination and composition measurement. Geometric mean SCC and TBC were 0.838 × 106 cells ml−1 and 581 × 103 cfu ml−1, respectively. Multivariable analyses revealed annual frequency of check-ups of milking system and total milk quantity per goat (among 53 variables) to be significant for increased SCC; no factor emerged (among 58 variables) to be significant for increased TBC. Negative correlation of SCC with total protein was found; mean total protein content in the bulk-tank milk in herds with SCC >0.75 × 106 cells ml−1 was 5.1% lower and in herds with SCC >1.5 × 106 cells ml−1, it was 7.8% lower.

In an age of flexible conditions about mandatory milk pasteurisation, this opinion-based research reflection supports the view that the knowledge and the awareness of milk-borne infections are key requirements to decrease the risks associated with raw milk. Providing an analysis of the current potential risks related to consumption of raw milk and raw milk products, we discuss the main reasons to continue to be vigilant about milk-borne pathogens and the current scenario in relation to the formal and clandestine sale of raw milk. Finally, we select some highly effective strategies to reduce the risks associated with raw milk in food services. Regardless of whether a country regulation allows or prohibits the trade of raw milk and its products, this is not the time to be negligent.

This research communication relates to the hypothesis that the consumption of raw or unprocessed cow's milk contributes to lowered prevalence of allergies. Thermal pasteurization of bovine milk can result in denaturation of minor whey proteins and loss of their bioactivity. Denaturation of bovine serum albumin (BSA), immunoglobulin G (IgG) and lactoferrin (LF) in skim milk was studied under different temperature (72, 75 or 78°C) and time (0–300 s) combinations. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) results revealed that denaturation of all 3 proteins occurred at 72°C and progressed with increase in temperature and holding time. About 59% of LF and 12% of IgG denatured under high-temperature short-time (72°C/ 15 s) pasteurization, while BSA was least impacted. To assess modulation of milk immunogenicity, secretion of selected T helper (Th)-type cytokines by human peripheral blood mononuclear cells (PBMCs) was studied in vitro in response to different concentrations of BSA (0.4–1.0 mg/ml) and IgG (0.8–1.6 mg/ml) in unheated skim milk. Addition of IgG at 1.6 mg/ml induced a prominent Th1-skewed cytokine profile that may not trigger a Th2-skewed allergic reaction. BSA did not appear to modulate milk immunogenicity to any significant extent.

Porungo is a traditional pasta filata cheese produced using raw milk throughout the southwest region of the state of São Paulo, Brazil. The objectives of this Research Communication were to evaluate the quality of raw milk used to make Porungo cheese, to characterize its production process, and to determine its chemical composition. The results showed that the raw milk met both Brazilian and international quality requirements. Chemically, Porungo can be classified as a medium to full fat semi-hard fresh cheese. Our study has allowed the first standards and regulations for Porungo to be established in Brazil. By virtue of this, the local producers are able to formalize their activity while consumers can have access to a safe and certified product.

In December 2016, Public Health England investigated an outbreak of campylobacteriosis in North West England, with 69 cases in total. Epidemiological, microbiological and environmental investigations associated the illness with the consumption of unpasteurised cows' milk from Farm X, where milk was predominantly sold from a vending machine. Campylobacter was detected in milk samples which, when sequenced, were identical in sequence type as pathogens isolated from cases (Clonal Complex ST-403, Sequence Type 7432). The farm was served with a Hygiene Emergency Prohibition Order to prevent further cases. To our knowledge, this is the first outbreak of campylobacter associated with unpasteurised milk in England since 1996. Our findings highlighted several important lessons, including that the current testing regime in England for unpasteurised milk is not fit for purpose and that the required warning label should include additional wording, underscoring the risk to vulnerable groups. There has been a substantial increase in both the volume of unpasteurised milk consumed in England and the use of vending machines to sell unpasteurised milk over the last 10 years, making unpasteurised milk more readily accessible to a wider population. The evidence generated from outbreaks like this is therefore critical and should be used to influence policy development.

Refrigerated storage of raw milk is a prerequisite in dairy industry. However, temperature abused conditions in the farming and processing environments can significantly affect the microbiological quality of raw milk. Thus, the present study investigated the effect of different refrigeration conditions such as 2, 4, 6, 8, 10 and 12 °C on microbiological quality of raw milk from three different dairy farms with significantly different initial microbial counts. The bacterial counts (BC), protease activity (PA), proteolysis (PL) and microbial diversity in raw milk were determined during storage. The effect of combined heating (75 ± 0·5 °C for 15 s) and refrigeration on controlling those contaminating microorganisms was also investigated. Results of the present study indicated that all of the samples showed increasing BC, PA and PL as a function of temperature, time and initial BC with a significant increase in those criteria ≥6 °C. Similar trends in BC, PA and PL were observed during the extended storage of raw milk at 4 °C. Both PA and PL showed strong correlation with the psychrotrophic proteolytic count (PPrBC: at ≥4 °C) and thermoduric psychrotrophic count (TDPC: at ≥8 °C) compared to total plate count (TPC) and psychrotrophic bacterial count (PBC), that are often used as the industry standard. Significant increases in PA and PL were observed when PPrBC and TDPC reached 5 × 104 cfu/ml and 1 × 104 cfu/ml, and were defined as storage life for quality (SLQ), and storage life for safety (SLS) aspects, respectively. The storage conditions also significantly affected the microbial diversity, where Pseudomonas fluorescens and Bacillus cereus were found to be the most predominant isolates. However, deep cooling (2 °C) and combination of heating and refrigeration (≤4 °C) significantly extended the SLQ and SLs of raw milk.

Children may be at higher risk for carriage of antimicrobial-resistant bacteria because of higher usage of antimicrobials. They also have higher rates of Shiga toxin-producing Escherichia coli (STEC) infections than other population groups. Some infections, particularly in children, are asymptomatic, but still lead to the excretion of large numbers of bacteria and viruses that may cause clinical disease in other individuals. That is one reason why, in Lower Saxony as in other German federal states – asymptomatic carriers of STEC are excluded from nurseries and schools until three consecutive stool samples test negative in order to prevent secondary cases. The prevalence of children who are asymptomatic STEC carriers is unknown. But if it is high, this measure would have substantial socioeconomic effects on families. Infections with extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBL-E) are an increasing problem for public health, especially for hospitals. However, there are no reliable estimates of the prevalence of asymptomatic ESBL-E carriers in Lower Saxony, as there is no mandatory requirement to report these carriers. In order to discuss the exclusion policies for children attending nurseries and ascertain a baseline of ESBL-E carriers, we conducted a cross-sectional study. The aim was to determine the prevalence of ESBL-E and STEC and identify risk factors for carriage in nursery children without diarrhoea (asymptomatic) aged 0–6 years in four selected districts in Northern Germany. During April–September 2014, we collected stool specimens with the support of voluntarily participating nurseries. We tested for STEC by PCR and for ESBL-E on chromogenic agar. Questionnaires answered by parents contained data on eating and drinking habits, outdoor activities, prior antibiotic treatment and animal contact for each participating child. We compared the epidemiological characteristics of ESBL-E carriers vs. non-carriers by using univariable analysis (P value, odds ratio and 95% confidence interval). We could not perform a statistical analysis for STEC carriers due to the low numbers of positive STEC specimens. Of 224 asymptomatic nursery children, we found a prevalence of 2·3% for ESBL-E carriage and 0·5% for STEC carriage. Asymptomatic ESBL-E carriers were more likely to have consumed raw milk, have had contact with pet rodents, or to have taken antibiotics during the preceding 6 months. We also found a high proportion of raw milk consumption (11%). We suggest that the low STEC prevalence in asymptomatic children supports the current practice of excluding STEC carriers from nurseries. The association between ESBL-E carriage and raw milk consumption and contact with pet rodents needs further investigation.

Epidemiological studies have shown an association between the consumption of raw farm milk and reduced incidence of allergy. In the present study, we fed untreated raw milk, gamma-sterilised milk, heat-treated milk or water to mice and compared their responses to allergen exposure and challenge treatment in a mouse model of gastrointestinal allergy. From weaning (3 weeks old), groups of BALB/c female mice (n 8) received raw milk, gamma-sterilised milk, heated milk or water via drink bottles, with the control group receiving water. All mice were fed a standard (dairy protein-free) rodent diet. At 6 and 8 weeks, groups were given intra-peritoneal injections with ovalbumin (OVA)/alum to sensitise them to the antigen. Controls were sham immunised. At week 10, mice were fasted and challenged four times on alternate days by intra-gastric administration with 50 mg OVA or saline. Levels of bacteria and milk proteins were assessed in milk samples. Mouse serum levels of specific IgE, IgG1 and IgG2a antibodies and mouse mast cell protease-1 (MMCP-1) were determined. Cytokine responses to 48 h activation with OVA were measured in cultured splenocytes from mice. Sterilised and heated milks contained no viable bacteria and reduced detectable levels of many milk proteins, in contrast to raw milk. Mice drinking raw milk had highest serum MMCP-1 and specific-OVA IgE responses. Cultured splenocytes from OVA-primed mice produced similar levels of IL-4 in response to the antigen; however, IL-10 levels were highest from mice drinking raw milk. Overall, the present study adds to the evidence that consuming different types of milk can affect allergic responses to a non-related dietary antigen.

Late blowing, caused by the outgrowth of clostridial spores present in raw milk and originating from silage, can create considerable product loss, especially in the production of hard and semi-hard cheeses. The conventional method for the isolation of Clostridium spp. from cheeses with late-blowing symptoms is very complicated and the identification of isolates is problematic. The aim of this work was the development of a multiplex PCR method for the detection of the main dairy-related clostridia such as: Cl. beijerinckii, Cl. butyricum, Cl. sporogenes, Cl. tyrobutyricum. Samples derived from silage, raw milk and hard cheese were analysed by the most probable number (MPN) enumeration. Forty-four bacterial strains isolated from gas positive tubes were used to check the reliability of the multiplex PCR assay. The specificity of the primers was tested by individually analysing each primer pair and the primer pair combined in the multiplex PCR. It was interesting to note that the samples not identified by the multiplex PCR assay were amplified by V2–V3 16S rRNA primer pair and the sequencing revealed the aligned 16S rRNA sequences to be Paenibacillus and Bacillus spp. This new molecular assay provides a simple promising alternative to traditional microbiological methods for a rapid, sensitive detection of clostridia in dairy products.

Mastitis is an inflammation of the mammary glands and in most cases it is caused by the presence of microorganisms. High mastitis rates in dairy cattle herds can cause an increase in total microorganism counts of bulk tank milk. The present paper was aimed at verifying whether the occurrence of mastitis in dairy cattle herds is reflected in raw-milk indicators of hygienic-sanitary quality. To observe the correlation among the analysed variables, we performed a logarithmical transformation (log10) of different indicator counts of raw milk and compared them with the occurrence of mastitis in dairy cattle herds. Few correlations were observed among mastitis cases in dairy cattle herds and the raw-milk indicators of hygienic-sanitary quality. We observed a negative correlation between the log10 of mesophilic aerobic plate counts and psychotropic aerobic plate counts when compared with the occurrence of no bacterial growth. The log10 of thermophilic aerobic plate counts and yeasts and mould aerobic plate counts presented a positive correlation with the cases of infectious mastitis and mastitis caused by Staphylococcus spp.

Raw whole milk of high microbial quality ([les ]4×104 cfu/ml) was processed using a ~15 l/h homogeniser with a high pressure (HP) valve immediately followed by cooling heat exchangers. The effects of homogenisation between 100 and 300 MPa (HP valve) with an initial milk temperature Tin=4 °C or 24 °C was investigated on the inactivation of: (i) endogenous alkaline phosphatase (ALP); (ii) endogenous milk flora and (iii) two Gram positive (Listeria innocua and Micrococcus luteus) and one Gram negative (Pseudomonas fluorescens) strains inoculated into milk. Temperatures T1 and T2 measured before and immediately after the HP valve, and fat globule size distributions were also determined. ALP activity slightly decreased after homogenisation above 250 MPa when Tin=4 °C (corresponding T2>58 °C), but markedly decreased above 200 MPa when Tin=24 °C (T2>60 °C). In contrast to inactivation induced by continuous short-time thermal treatments, ALP inactivation induced by HP homogenisation was clearly due to mechanical forces (shear, cavitation and/or impact) in the HP valve and not to the short ([Lt ]1 s) residence time at temperature T2 in the same valve. Inactivation of the three exogenous microorganisms led to similar conclusions. Homogenisation at 250 MPa or 300 MPa (Tin=24 °C) induced a 2–3 log cycle reduction of the total endogenous milk flora and a 1·5–1·8 log cycle reduction of inoculated List. innocua. Higher reduction ratios (2–4 log cycles) were obtained for the two other microorganisms. The highest levels of ALP inactivation corresponded to the highest extents of microbial reduction. Running the milk twice or three times through the homogeniser (recycling), keeping temperature T1≈29 °C and pressure=200 MPa, increased homogenisation efficiency.

Difficulties with the analysis of opaque milk and dairy products can be resolved by rendering the sample transparent. This allows direct photometric measurements of the reaction mixtures, and all sample pre-treatment steps become unnecessary. For this, several reagents have been reported in literature. Among them, the ‘Clarifying Reagent’ is mainly described here: its composition is given because it is now released from patenting restrictions. Its main property is a low absorbance between 340 and 800 nm. So it can be used in many chemical and enzymatic colorimetric measurements for estimating the quality of milk and dairy products.

Psychrotrophs, particularly strains of Pseudomonas fluorescens, dominate the microflora of refrigerated raw milk and secrete heat-stable extracellular enzymes (proteases and lipases) which survive pasteurisation and even UHT heat treatments and degrade the casein and fat components of raw milk causing a reduction in cheese yield, gelation of UHT milk and off flavours in many dairy products. These enzymes are usually produced in the late log/early stationary growth phases when the cell density is high. This fact indicated that induction of these enzymes may be a candidate for quorum sensing, a phenomenon by which bacteria can sense and respond to cell population size by means of chemical signals based on the homoserine lactone molecule. It is assumed that the production of these enzymes contributes to the selective advantage of psychrotrophs and hence have a significant effect on their growth kinetics. In the work reported here nine proprietary homoserine lactone compounds were screened, using water as a control, for their effect on the lag phase duration (LPD) and exponential growth rate (EGR) of three strains of Pseudomonas fluorescens isolated from refrigerated raw milk after 1, 3 and 5 days storage. Two compounds viz. N-benzoyloxycarbonyl-L-homoserine lactone and N-3-oxyhexanoyl-DL-homoserine lactone were found to significantly (P<0·001) reduce the LPD and increase the EGR of the three strains. Further work with these compounds is warranted, monitoring growth in parallel with enzyme production, to determine the extent to which extracellular enzyme production is mediated by the quorum sensing phenomenon in this species.

The effect of the hygienic quality of milk on changes in microbial counts and biogenic amine content was evaluated during ripening of goat cheeses manufactured from pasteurized and raw milks at 1, 14, 30, 60 and 90 d. The original milk, rennet, curd and whey were also included in the study. The pH, salt content and extent of proteolysis in the cheese were also evaluated. Spermidine and spermine were the main amines in raw milk, while they were minor amines in cheeses. Other amines increased markedly during ripening, tyramine being the main amine in cheese made from raw milk and cadaverine and putrescine in those produced from pasteurized milk. Enterobacteriaceae counts decreased during ripening whereas those of lactic acid bacteria increased, especially lactobacilli and enterococci. Cheese made from raw milk showed higher microbial counts during ripening than those made from pasteurized milk, especially for Enterobacteriaceae and enterococci, counts being 2 or 3 log units higher. Raw milk cheese showed remarkably higher biogenic amines compared with pasteurized milk cheeses. Therefore, pasteurization of milk causes a decrease in final biogenic amine content of cheese as a result of the reduction of its microbial counts.

Manchego cheese can be manufactured from raw or pasteurized ewes' milk. An automatic purge and trap apparatus, coupled to a GC-MS was used to isolate, identify and compare the relative amounts of the volatile components of raw and pasteurized Manchego cheese during ripening. The majority of volatile compounds were more abundant in raw milk (RM) cheeses than in pasteurized milk (PM) cheeses. Alcohols and esters predominated in the profile of RM Manchego cheese, while methyl-ketones and 2,3-butanedione were quantitatively important in PM cheeses. Branched chain alcohols were much more abundant in RM cheeses. The discriminant analysis separated 100% samples into RM or PM cheeses by using only 16 volatile compounds. Aroma intensity was correlated with esters, branched chain aldehydes and branched chain alcohols in RM cheeses, and with esters, branched chain aldehydes, 2-methyl ketones and 2-alkanols in PM cheeses. Diacetyl was positively correlated with the aroma attribute ‘toasted’ and negatively correlated with aroma quality in PM cheeses.

The ability of Listeria monocytogenes to grow at low temperatures and increase to hazardous levels during storage of dairy products is of great concern to the dairy industry. List. monocytogenes was detected in 3·6% of 774 cows' milk samples from 114 farms in Central Spain (Gaya et al. 1998). Ewes' and goats' milks produced in Spain had contamination levels of 2·2 and 2·6% respectively (Rodríguez et al. 1994; Gaya et al. 1996). List. monocytogenes may survive during the manufacture and ripening of most cheese varieties, with higher risk in soft surface-ripened cheeses owing to the rise in pH during ripening (Maisnier-Patin et al. 1992; Giraffa & Carminati, 1997).

The efficacy with which actively growing bacteriocin-producing cultures inhibit List. monocytogenes in some cheese varieties has been demonstrated. Nisin-producing starters inhibited List. monocytogenes V7 in Camembert cheese (Maisnier-Patin et al. 1992) and List. innocua BL86/26 in raw ewes' milk Manchego cheese (Rodríguez et al. 1998). A lacticin 3147-producing starter culture (McAuliffe et al. 1999) reduced List. monocytogenes Scott A levels in cottage cheese. Inoculation of milk with enterocin AS-48-producing Enterococcus faecalis INIA 4 inhibited List. monocytogenes strains Ohio and Scott A during the manufacture and ripening of Manchego cheese (Nuñez et al. 1997).

Semi-hard cheese varieties from raw or pasteurized cows' milk mixed with different proportions of ewes' and/or goats' milk represent ∼ 50% of Spanish cheese production. The objective of the present work was to evaluate the antilisterial activity of three bacteriocin-producing lactic acid bacteria strains isolated from milk (Rodríguez et al. 2000) during the manufacture and ripening of a semi-hard cheese manufactured from raw milk.

Many psychrotrophic bacteria contaminating raw milk produce phospholipase that withstands pasteurization and UHT treatments. This enzyme acts on the milk fat globule membrane and exposes triacylglycerides to the action of lipase. Phospholipase production by various isolates of Pseudomonas was investigated. The isolates were cultured aerobically at 8 °C in nutrient broth, McKellar's minimal salts medium, Chrisope's medium, and skim milk. Each strain produced phospholipase during the 50 h incubation. Enzyme production varied significantly (P < 0·001) with strain and growth medium. Strains varied significantly (P < 0·001) in their enzyme production in each medium and during the incubation time as well. Strain, incubation time, and the growth medium significantly influenced (P < 0·001) heat stability of the enzyme activity. Pasteurization reduced the activity, but did not eliminate it in skim milk.