The effect of intramammary inoculation of Lactobacillus perolens CRL 1724 on bovine udders at drying off was evaluated through histological examination of the canal and cistern tissues. The persistence of the strain in the udder 7 d post inoculation was also determined. Lb. perolens CRL 1724 was recovered from all mammary quarters and no clinical signs or teat damage were observed after inoculation of 106 cfu/ml. The udders showed a normal structural aspect and there were no modifications of the milk appearance. Lb. perolens CRL 1724 cells were evidenced on the surface of the epithelial cells of the cistern without causing any morphological modifications or cell alterations. Lb. perolens CRL 1724 produces a mild inflammatory reaction, characterized by recruitment of neutrophils to the epithelial zone and a slight hyperaemia into blood vessels. This preliminary study provides important information for further studies directed towards the inclusion of Lb. perolens CRL 1724 in the design of probiotic products for preventing bovine mastitis in non-lactating dairy cows.

The effects of a bovine whey peptide product enriched in proline (wPRP) on the solubility of milk proteins were tested under ambient conditions or following heat treatment at 75 and 100 °C, for 1 and 15 min, followed by post-incubation storage at either ambient temperature or 4 °C for up to 7 d. wPRP promoted solubilisation of milk proteins in a concentration-dependent manner without heat treatment and also after heat treatment at 75 and 100 °C, and the effect was enhanced after storage under either ambient or refrigerated storage conditions. Interactions of wPRP and milk proteins were monitored by particle size analysis and tryptic digestion and specifically linked with solubilisation of αS1 casein (αS1-Cn), which supported observed changes in milk protein solubility. The results suggested that wPRP preferably prevented or reversed physical versus covalent protein aggregation, with the relaxation of hydrophobic interactions at 4 °C providing an additive effect. This application of wPRP represents a novel approach to stabilisation of dairy proteins following thermal processing with industrial usefulness yet to be explored.

The purpose of this work was to analyse the influence of rennet from different Cynara cardunculus plants, selected for its clotting and proteolytic activity on caseins, on the characteristics of manufactured ‘Torta del Casar’ cheeses. After classifying the cardoon according to proteolytic activity into five groups of greater or lesser activity, 16 batches of cheeses were made with rennet derived from different wild cardoon plants. We observed a major development of the proteolysis during ripening leading to the generation of non-protein nitrogen compounds. Especially noteworthy was the relationship of amino acid nitrogen (AN) generation with rennet clotting activity after 24 h of maceration, and the fact that the production of biogenic amines was not related to the proteolytic activity of the rennet. The activities of the rennet observed ‘in vitro’ were also developed ‘in vivo’ in the cheeses, with the different rennets used affecting the final sensory characteristics of cheeses. The rennet with high clotting activity after 24 h of maceration was positively correlated with the creaminess, viscosity, and acceptability of the cheese. However, the high proteolytic activity rennet negatively influenced the acidity, bitterness, and creaminess parameters. Therefore the most appropriate cardoons for making this cheese are those with higher clotting activities and moderate proteolytic activities especially on β-casein. The use of controlled and characterised cardoons in the manufacturing process of Torta del Casar is fundamental to obtaining the homogeneous product demanded by the Torta del Casar Registry of the Protected Designation of Origin.

In this work, the genetic variation of milk FA was investigated in three different bovine breeds, the Jersey, the Piedmontese and the Valdostana, and at different lactation stages. All animals were genotyped for 21 Single Nucleotide Polymorphisms located within nine candidate genes involved in lipid synthesis: diacylglycerol acyltransferase 1 and 2 (DGAT1, 2); stearoyl-CoA desaturase (SCD); growth hormone receptor (GHR); fatty acid synthase (FASN); acyl-CoA dehydrogenase (ACAD); fatty acid binding protein (FABP4); lipoprotein lipase (LPL); and leptin gene (LEP). The highest milk-fat Jersey breed also showed the highest content of saturated FA. Throughout lactation, the breeds showed a similar variation in the FA, with a decrease in the short-chain, this was accompanied by a general increase in the long chain FA at the end of lactation. The increase in long chain saturated FA was particularly evident in the case of the Jersey. The effect of SCD gene on the C14 desaturation index was confirmed; the DGAT1 gene was polymorphic only in the Jersey breed, but its effect was confirmed only on milk fat content; three further potential candidate genes were identified: first, the FABP4 gene, which was found to influence medium and long chain FA in all the breeds, but not the desaturation indices; second, the FASN gene, which was found to influence the amount of PUFA in the Piedmontese and the Valdostana, and third, the LPL gene, which was found to affect fat content in the Piedmontese.

Considerable inhibitory antibacterial actions were exerted by the soybean 11S subunit comparable with nisin on the proliferation of total viable count, Pseudomonas count and Enterobacteriaceae count in bovine milk stored at 4 or 25 °C for 30 d and 48 h, while 7S and lysozyme were much less effective. The maximum magnitudes of bacterial reduction by 11S and nisin were in the range 2–4 log CFU/ml. The proliferation of 3 pathogenic bacteria (Listeria monocytogenes, Salmonella Enteritidis and Escherichia coli O157:H7) artificially inoculated into raw milk stored at 4 or 25 °C were particularly and significantly (P<0·05) reduced by 11S subunit and nisin (0·5% w/v), but only slightly by 7S and moderately by lysozyme. Lactose consumption, acidity development and casein degradation during storage of bovine raw milk were attenuated during storage at 4 or 25 °C and sensorial traits were better maintained by supplementation with 11S (0·5% w/v). 11S subunit may be used a safely food preservative, if permitted.

The objective of this study was to examine the quarter health status of quarter-individually and conventionally milked cows. The MultiLactor®, a quarter-individual milking system (MULTI), has single guided tubes which provide milking on the quarter level with a low system vacuum level (37 kPa), sequential pulsation and periodic air inlet. The conventional milking system (CON) was equipped with a milking cluster where the system vacuum level was adjusted to 40 kPa. A total of 84 German Holstein cows, randomly divided into two groups, were included in the study. Over a period of 32 trial weeks, quarter foremilk samples were taken every week to determine somatic cell count (SCC). Bacteriological examinations and udder palpation were conducted at three different times. During the trial period, median SCC of quarter foremilk samples in both groups did not exceed the threshold value of 100 000 cells/ml. The results of the F test showed that the milking system (P = 0·0587) and days in milk (DIM) (P = 0·8066) had no significant effects on the quarter health status. On the other hand, lactation (P = 0·0396), quarter health status in the previous week (P < 0·0001) and trial week (P = 0·0061) affected quarter health status significantly. The estimated probabilities of the occurrence of a suspicious quarter (SCC > 100 000 cells/ml) were 19·97% (CON) and 31·72% (MULTI). However, the test of differences in the Least Square Means (LSM) showed no significant differences (P = 0·0585) between CON and MULTI. The estimated probability of quarters becoming suspicious during the first lactation was 12·51% for both groups. With an increasing number of lactation, the probability of a quarter becoming suspicious clearly increased (2nd lactation: 32·73% and 3rd lactation: 36·19%). The results also showed that the percentage of quarters with bacteriological findings revealed a stronger increase over time for MULTI than for CON.

Agriculture and animal husbandry are important contributors to global emissions of greenhouse (GHG) and acidifying gases. Moreover, they contribute to water pollution and to consumption of non-renewable natural resources such as land and energy. The Life Cycle Assessment (LCA) methodology allows evaluation of the environmental impact of a process from the production of inputs to the final product and to assess simultaneously several environmental impact categories among which GHG emissions, acidification, eutrophication, land use and energy use. The main purpose of this study was to evaluate, using the LCA methodology, the environmental impact of milk production in a sample of 41 intensive Italian dairy farms and to identify, among different farming strategies, those associated with the best environmental performances. The functional unit was 1 kg Fat and Protein Corrected Milk (FPCM). Farms showed characteristics of high production intensity: FPCM, expressed as tonnes per hectare, was 30·8±15·1. Total GHG emission per kg FPCM at farm gate was 1·30±0·19 kg CO2 eq. The main contributors to climate change potential were emissions from barns and manure storage (50·1%) and emissions for production and transportation of purchased feeds (21·2%). Average emission of gases causing acidification to produce 1 kg FPCM was 19·7±3·6 g of SO2 eq. Eutrophication potential was 9·01±1·78 ${\rm PO}_{\rm 4}^{{\rm 3} -} {\rm eq}.$ per kg FPCM on average. Farms from this study needed on average 5·97±1·32 MJ per kg FPCM from non-renewable energy sources. Energy consumption was mainly due to off-farm activities (58%) associated with purchased factors. Land use was 1·51±0·25 m2 per kg FPCM. The farming strategy based on high conversion efficiency at animal level was identified as the most effective to mitigate the environmental impact per kg milk at farm gate, especially in terms of GHG production and non-renewable energy use per kg FPCM.

This study investigated factors affecting milk production and lactation curves from complete lactations of Lacaune dairy sheep. Animals were part of a single flock under intensive management and were milked twice daily starting at lambing. The results of the analyses of 7788 complete lactations showed an average total milk yield of 434±183 l from lactations 234±63 d long, with an average lambing interval of 302±65 d. A Pollott additive mathematical model was used to estimate complete lactation curves. Clustering analysis identified four lactation types among Lacaune dairy sheep differing mainly in productivity i.e. milk yield per lactation (MY) and length of lactation (DIM). The so-called SL type involved short, less productive lactations (n=2137; 27·4%; MY=222±75·5 l and DIM=182±52·9 d). The SN type involved short lactations of normal productivity (n=2039; 26·2%; MY=396±73·7 l and DIM=205±33·1 d). The LP type involved long and productive lactations (n=2169; 27·9%; MY=487±70·5 l and DIM=265±40·7 d), while the LVP type included long and extremely productive lactations (n=1443; 18·5%; MY=694±114·0 l and DIM=295±54·7 d). Sheep showing the best lactation curves were usually younger than other sheep, and they had higher yield during the previous lactation, a shorter previous dry period (55±50·4 for LP and 61±55·0 d for LVP types) and longer lambing intervals. In addition, they tended to be born in September and to lamb in March, October and December. Sheep were remarkably stable in their lactation curve behaviour: the curve type observed for the first lactation was highly likely to persist in subsequent lactations (P<0·0001). These results suggest that farmers can use the shape of the first lactation curve to guide their selection of ewes for breeding and retention on the farm, thereby improving flock productivity.

High fat dairy products, such as butter and margarine can be contaminated during the milk production process with a residue called Trichloromethane (TCM), which results from the use of chlorine based detergent solutions. Although, TCM concentrations in Irish products are not at levels that are a public health issue, such contamination can cause marketing difficulties in countries to which Irish products are being exported. In an attempt to reduce such milk residues, a template procedure was developed, tried and tested on 43 farms (from 3 processing companies). This involved identifying farms with high TCM milk, applying corrective action in the form of advice and recommendations to reduce TCM and re-measuring milks from these farms. Trichloromethane in milk was measured by head-space gas chromatography with electron capture detector. The TCM reduction strategy proved successful in significantly reducing the levels in milk in the farms tested, e.g. TCM was reduced from 0·006 to the target of 0·002 mg/kg (P < 0·05). The strategy was then applied to farms who supplied milk to six Irish dairy processors with the objective of reducing TCM in those milks to a level of ⩽0·002 mg/kg. Initially, milk tankers containing milks from approximately 10–15 individual farms were sampled and analysed and tankers with high TCM (>0·002 mg/kg) identified. Individual herd milks contributing to these tankers were subsequently sampled and analysed and farms supplying high TCM identified. Guidance and advice was provided to the high TCM milk suppliers and levels of TCM of these milk supplies were monitored subsequently. A significant reduction (minimum P < 0·05) in milk TCM was observed in 5 of the 6 dairy processor milks, while a numerical reduction in TCM was observed in the remaining processor milk.

The objective was to identify predisposing factors for increased risk of involuntary culling in adult Holstein–Friesian dairy cows. Data were sourced from Scotland's Rural College (SRUC) Dairy Research Centre. Between September 2003 and August 2010 175 cows were culled, a herd culling rate of 33·7%. The major reasons for involuntary culling were fertility (27·4%) and udder problems (26·9%). In the analysis, the culled cows were matched with their cohorts that survived to a later lactation. To identify predisposing factors, a binary logistic model was applied. Cows with higher than average body condition score (BCS) at last service were five times [Odds Ratio (OR) = 4·8] more likely to be culled due to infertility. Cows with low protein yield on day 60 ± 5 in lactation were ten times less likely (OR = 0·1) to be culled. In first lactation heifers, only BCS at last service increased the risk of involuntary culling due to infertility (OR = 13·0). A high milk yield acceleration was a significant (P = 0·04) factor in increasing the risk, five times (OR = 5·2) more, of culling cows due to udder problems. In conclusion, a high BCS at last service, high milk protein yield at around day 60 in lactation and acceleration of milk yield after calving exposed cows to a risk of being culled involuntarily. In practice, monitoring of traits that indicate metabolic imbalance could assist identifying cows at high risk of being culled and contribute to reducing the associated risk through a more effective timely decision.

A randomised clinical trial was conducted to explore the effect of orally supplemented medium-chain fatty acids (MCFA) to heifers and cows starting 6–8 weeks prior to expected calving date on blood and milk polymorphonuclear neutrophilic leucocyte (PMNL) apoptosis between 1 and 3 d in milk (DIM). The effects of MCFA-supplementation on the likelihood of intramammary infections (IMI) in early lactation, and test-day somatic cell count (SCC) and average daily milk yield (MY) during the first 4 months of lactation were evaluated as well. Twenty-two animals were included of which half were orally supplemented with MCFA starting 6–8 weeks prior to calving and half served as non-supplemented controls. The PMNL viability in both blood and milk was quantified using dual-colour flow cytometry with fluorescein-labelled annexin and propidium iodide. In non-supplemented animals, % blood PMNL apoptosis significantly increased between start of supplementation and early lactation, reflecting a potential reduction in innate immune capacity, whereas this was not true in the MCFA-supplemented animals. Similar results were seen in milk PMNL apoptosis. Overall, the % apoptotic milk PMNL between 1 and 3 DIM was significantly lower in the MCFA-supplemented group compared with the non-supplemented group. There was no substantial effect of oral MCFA-supplementation on the likelihood of quarter IMI nor on the composite test-day milk SCC or average daily MY. In conclusion, oral MCFA-supplementation starting 6–8 weeks before expected calving date supported the blood and milk neutrophil viability in early lactating dairy cows. Still, this was not reflected in an improvement of udder health nor MY in early and later lactation. The results should trigger research to further unravel the mechanisms behind the observed immunomodulating effect, and the potential relevance for the cows' performances throughout lactation.

The aim of this study was to evaluate the effect of feeding goats with distilled and non-distilled thyme leaves (Thymus zygis subsp. gracilis) on the physicochemical composition and technological properties of pasteurised goat milk, and on the physicochemical composition, phenolic content, oxidative stability, microbiology, sensory and texture profile of Murcia al Vino goat cheese. One group of goats was fed the basal diet (control), the second and third groups were fed with different levels of distilled (10 and 20%) or non-distilled (3·75 and 7·5%) thyme leaves. Goat milk physicochemical composition was significantly affected by the substitution of 7·5% of basal goat diet with non-distilled thyme leaves (increase in fat, protein, dry matter and PUFA content), while goat milk clotting time was increased significantly by the introduction of 20% distilled thyme leaves, which reduces its technological suitability. Microbiology, sensory and texture profiles were not affected by the introduction of distilled thyme leaves. The introduction of distilled and non-distilled thyme leaves as an alternative feed to diet can lead to an inhibition of lipids oxidation. The introduction of distilled and non-distilled thyme leaves into goat's diet can be successfully adopted as a strategy to reduce feeding costs and to take advantage of the waste from the production of essential oils, minimising waste removing costs and the environmental impact.

Studies indicate a positive association between dietary salt intake and some diseases, which has promoted the tendency to reduce the sodium in foods. The objective of this study was to determine the equivalent amount of different sodium chloride replacements required to promote the same degree of ideal saltiness in butter and to study the sensory profile of sodium chloride and the substitutes using the analysis of Temporal Dominance of Sensations (TDS). Using the magnitude estimation method, it was determined that the potencies of potassium chloride, monosodium glutamate and potassium phosphate relative to the 1% sodium chloride in butter are 83·33, 31·59 and 33·32, respectively. Regarding the sensory profile of the tested salt substitutes, a bitter taste was perceived in the butter with potassium chloride, a sour taste was perceived in the butter with potassium phosphate and sweet and umami tastes were dominant in the butter with monosodium glutamate. Of all the salt substitutes tested calcium lactate, potassium lactate, calcium chloride and magnesium chloride were impractical to use in butter.

Genotypic and technological properties, antibiotic susceptibility and antimicrobial activity of 35 Leuconostoc strains, isolated from different Italian raw milk cheeses, were investigated. RAPD-PCR was used to study genetic variability and to distinguish closely related strains. The results showed a high degree of heterogeneity among isolates. All the strains had weak acidifying activity and showed low proteolytic and lipolytic activities. Reduction activity, was generally low. All the Leuconostoc were susceptible to ampicillin, mupirocin, erythromycin, quinupristin/dalfopristin and tetracycline. Many strains were classified as resistant to oxacillin, ciprofloxacin and nitrofurantonin, while all isolates were found resistant to vancomycin. PCR-based detection did not identify any of the common genetic determinants for vancomycin (vanA, vanB, vanC1, vanC2, vanC3, vanD, vanE, vanG) or erythromycin (ermB and ermC). Tetracycline resistance genes were detected in 25 tetracycline susceptible strains, the most frequent one being tetM. One strain, belonging to Ln. pseudomesenteroides species, was positive for the presence of the int gene of the Tn916/Tn1545 trasposon family. This is the first time the conjugative transposon Tn916 has been detected inside the Leuconostoc species. All strains showed antimicrobial activity against Enterococcus faecalis and Ent. faecium. The presence of genes encoding amino-acid decarboxylases (hdc and tdc) was not detected. Some strains are interesting in view of their use in cheese production as starter and non starter cultures.

As grain prices rise, the search for alternative glycogenic precursors in animal feed becomes increasingly important, and this study was conducted to determine if the replacement of starch with glycerol, as an alternative glycogenic precursor, affects the milk metabolic profile and milk coagulation ability, and therefore the quality of the milk. Eight primiparous mid-lactation Holstein cows were fed during a replicated 4 × 4 Latin square trial with four different isoenergetic rations: (1) control (T0) fed a total mixed ration (TMR) with barley meal; (2) group T1, decreased barley content, replaced isoenergetically with 1 kg crude glycerol; (3) group T2, the barley meal was replaced with 2 kg of crude glycerol; and (4) group T3 the barley meal was replaced with 3 kg of crude glycerol. Rumen, blood and milk samples were collected at the end of every 21-d treatment period. Rumen samples were analysed for proportion of total volatile fatty acid (VFA), blood samples for insulin and glucose, and milk for metabolites (e.g. citric-acid cycle compounds). The change in glycogenic precursors had a positive effect on rumen VFA proportions; the proportion of propionic acid increased (P < 0·001). Milk protein (P < 0·001) and curd firmness (P < 0·001) both increased. The increase in milk protein concentration may have been due to an increase in microbial protein. Regarding the milk metabolic profiles, different signals were positively associated with coagulation ability and change in the diet. Based on this study, changing the glycogenic precursor in animal diet in this way is possible, and may have no immediate deleterious consequences on milk quality or cow health. Indeed, there is evidence for benefits from this substitution.

Hispánico cheese, manufactured from a mixture of cow and ewe milk, is representative of cheese varieties made using milk from more than one animal species in Mediterranean countries. The shortage of ewe milk production in autumn hinders the uniformity of Hispánico cheese composition throughout the year. To surmount this inconvenience of ewe milk seasonality, curds made in spring from raw and pasteurized ewe milk were stored frozen and used four months later for the manufacture of Hispánico cheese. Experimental cheeses were made by mixing fresh curd from pasteurized cow milk with thawed curd from raw or pasteurized ewe milk, and control cheese from a mixture of pasteurized cow and ewe milk in the same proportion. Characteristics of experimental and control cheeses throughout a 60-d ripening period were investigated. On the one hand, the experimental cheese containing frozen curd from raw ewe milk showed the highest counts of staphylococci, Gram-negative bacteria and coliforms, the highest levels of aminopeptidase and esterase activity, and the highest concentrations of free amino acids, free fatty acids, alcohols and esters. On the other, the experimental cheese containing frozen curd from pasteurized ewe milk had concentrations of free amino acids, free fatty acids and volatile compounds similar to those of control cheese, with the only exception being a higher level of ketones. Flavour intensity reached the highest scores in the experimental cheese containing frozen curd from raw ewe milk, followed by the experimental cheese containing frozen curd from pasteurized ewe milk. Flavour quality scores of both experimental cheeses were similar, and lower than those of control cheese.

The objective of this study was to determine the effect of cow genetic strain on fatty acid (FA) profiles in adipose tissue and milk. Adipose samples from two subcutaneous (shoulder and tail-head) and three visceral (kidney channel, mesenteric and omental) depots were obtained post mortem from New Zealand (NZ; n = 8) and North American (NA; n = 8) Holstein–Friesian cows. At the time of slaughter cows were in similar body condition (NZ: 4·0 ± 0·03, NA: 4·0 ± 0·02; ±sd) and stage of lactation (NZ: 90 ± 11·2 d; NA: 83 ± 4·3 d; ±sd). Milk was collected during the a.m. milking prior to slaughter and milk fat was extracted. Adipose and milk fat FA were quantified using gas chromatography. NZ cows had a lower proportion of saturated FA in shoulder, tail-head and omental adipose tissue and a greater proportion of mono-unsaturated FA and an elevated Δ9-desaturase index in shoulder and tail-head adipose tissue. The proportions of individual FA differed between adipose depots, with proportions of de-novo FA greater in subcutaneous compared with visceral adipose depots. Milk from NZ cows contained greater concentrations of short chain FA (C8 : 0–12 : 0) and CLA, and less cis-9 18 : 1 than milk from NA cows. Regression analysis identified moderate associations between milk FA and shoulder adipose tissue FA for 18 : 2 (R2 = 0·24), 18 : 3 n − 3 (R2 = 0·39), and polyunsaturated fatty acids (R2 = 0·38). Results from this study support the hypothesis that genetic strain dictates FA profiles in adipose tissue and milk and may alter the metabolic status of the various adipose depots differently. The data further support the premise that genetic strain affects the metabolic status of the various adipose depots differently. Elucidating the mechanisms that regulate the different adipose depots in the NZ and NA strains will increase our understanding of tissue mobilization and replenishment.

The objective of this study was to collect and analyse milking data from a sample of commercial farms with swingover herringbone parlours to evaluate milking efficiency over a range of parlour sizes (12–32 milking units). Data were collected from 19 farms around the Republic of Ireland equipped with electronic milk metres and herd management software that recorded data at individual milking sessions. The herd management software on each farm was programmed to record similar data for each milking plant type. Variables recorded included cow identification, milking date, identification time, cluster-attachment time, cluster/unit number, milk yield, milking duration, and average milk flow rate. Calculations were performed to identify efficiency benchmarks such as cow throughput (cows milked per h), milk harvesting efficiency (kg of milk harvested per h) and operator efficiency (cows milked per operator per h). Additionally, the work routine was investigated and used to explain differences in the benchmark values. Data were analysed using a linear mixed model that included the fixed effects of season-session (e.g. spring-AM), parlour size and their interaction, and the random effect of farm. Additionally, a mathematical model was developed to illustrate the potential efficiency gains that could be achieved by implementing a maximum milking time (i.e. removing the clusters at a pre-set time regardless of whether the cow had finished milking or not). Cow throughput and milk harvesting efficiency increased with increasing parlour size (12 to 32 units), with throughput ranging from 42 to 129 cows/h and milk harvesting efficiency from 497 to 1430 kg/h (1–2 operators). Greater throughput in larger parlours was associated with a decrease in operator idle time. Operator efficiency was variable across farms and probably dependent on milking routines in use. Both of these require consideration when sizing parlours so high levels of operator efficiency as well as cow throughput can be achieved simultaneously. The mathematical model indicated that application of a maximum milking time within the milking process could improve cow throughput (66% increase in an 18-unit parlour when truncating the milking time of 20% of cows). This could allow current herd milking durations to be maintained as herd size increases.

A specific and simple competitive enzyme-linked immunosorbent assay (ELISA) was developed to determine bovine β-casein phosphopeptides (β-CPP) in casein phosphopeptides (CPP) or CPP complexes such as casein phosphopeptide amorphous calcium phosphate complexes added into dairy products. The method combines sample pretreatment designed for CPP enrichment and anti-β-CPP(f(1–25)) monoclonal antibody 1A5 (mAb 1A5). The mAb 1A5 bound specifically to the tryptic phosphopeptides from β-casein but not from αs1- or αs2-casein. Reactivity was also influenced by the extent of the phosphorylated form of serine residues. Based on the sequence-specific recognition and contribution of phosphorylated serine residues, the epitope of mAb 1A5 was found to reside within the cluster motif Ser(P)-Ser(P)-Ser(P)-Glu-Glu and the surrounding residues in β-CPP. The competitive ELISA developed here can be used as an alternative to specialised and expensive techniques such as mass spectrometry. In particular, it is suitable for the measurement of CPP or CPP complexes in dairy products, which contain closely related endogenous molecular species.

Caseinomacropeptide (CMP), a 7-kDa phosphoglycopolypeptide fragment released from κ-casein during milk renneting, is heterogeneous with respect to post-translational glycosylation. Several studies have reported that CMP has growth-promoting activity on lactic acid bacteria belonging to the genera Bifidobacterium. The aim of this study was to evaluate the effect of glycosylation and sequence variations between bovine and caprine CMP on the growth of two probiotics: Lactobacillus rhamnosus RW-9595-M and Bifidobacterium thermophilum RBL67. The growth-promoting activities of CMP (mixture of glycosylated (gCMP) and non-glycosylated (aCMP) fractions), aCMP and gCMP were measured in a basal minimal culture medium using turbidimetric microplate assay at 37 °C. Supplementation of the culture media at 2 mg/ml significantly improved maximum growth by 1·5 to 1·8 times depending on the strain, the additive (CMP, aCMP, gCMP), and the bovine or caprine origin (P < 0·05). CMP preparations also decreased the time needed to reach the inflexion point of the growth curve and increase the cell density at that time (P < 0·05). The effects of CMP preparations were dose dependent and significantly superior to the effect of bovine β-lactoglobulin added to the culture media. As gCMP and aCMP were as efficient as bovine and caprine CMP (P > 0·1), it was concluded that the presence of oligosaccharides linked to CMP was not essential for growth-promoting activity of CMP.