Mixed cultures of different EPS-producing Streptococcus thermophilus strains in combination with a Lactobacillus delbrueckii subsp. bulgaricus strain with negligible EPS-production were used for yoghurt production. The yoghurt texture was characterised with respect to sensory, rheological and microstructural properties and the EPS-concentrations were determined. The cultures resulted in yoghurts with highly different texture properties, and positive interactions between certain Streptococcus thermophilus strains were observed. The underlying properties of yoghurt texture are multidimensional, but a number of microstructural characteristics were apparent in the yoghurts with the highest mouth thickness, creaminess and viscosity. A strong protein network, not too dense and with medium size pores containing EPS, seems associated with these properties. The presence of capsular polysaccharides (CPS) also appeared to be beneficial as did a combination of EPS types, which were distributed differently in the protein network (in serum pores, respectively in association with protein). Obviously, a certain concentration of EPS must be present to provide for these effects on yoghurt texture, but other factors than concentration per se seem more important.

Postpartum production performance of dairy heifers may be enhanced by prepartum milking by alleviating the stress of the periparturient period. The objective of this study was to determine the impact of prepartum milking of dairy heifers on postpartum reproduction, udder health, milk production and other associated production characteristics. Pregnant heifers (Holstein, n=21; Jersey n=10) were assigned to either a prepartum milked (premilked, n=15) or control (n=16) group. Premilked heifers were milked twice daily starting 3 weeks prior to anticipated calving dates, and milk yield recorded at each milking. All heifers were evaluated on days 21, 14 and 7 before calving, and udder oedema scores and milk conductivity readings were recorded. Following calving, measurements were taken twice weekly to assess udder oedema, milk conductivity (indicative of udder infection), and reproductive health, which included palpation for uterine tone and uterine position, vaginal electrical impedance (VEI) and the quantification of cross-sectional area of the uterine horns (uterine difference) by transrectal ultrasonography. Uterine tone, uterine position, uterine difference, and VEI did not differ (P>0·10) with treatment. Overall, up to week six inclusive, postpartum premilked heifers had lower (P<0·01) udder oedema scores than control heifers and up to week five inclusive, had lower (P<0·01) milk conductivity readings (indicative of fewer incidences of udder infections) than control heifers. The premilked heifers of both breeds produced more milk (P<0·01) at calving and more milk overall from calving to day 60 postpartum than the control heifers. In summary, udder health and milk production were improved post calving in premilked heifers compared with controls. However, no overt differences in reproductive characteristics were observed between the premilked and control heifers.

The major whey proteins IgG, serum albumin and α-lactalbumin were purified from camel milk using gel permeation and ion-exchange chromatography. Specific antisera against each of them were raised and used to quantify their heat denaturation in early or mature milk over a range of 60–90 °C for 10–60 min using the single radial immunodiffusion technique. The heat denaturation midpoints for the mature milk heated 30 min were 67·2, 73·0 and 77·5 °C for IgG, albumin and α-lactalbumin respectively. The early milk was more sensitive to heat treatment with coagulation at low temperature and heat denaturation midpoints of 64·8, 71·6 and 72·6 °C respectively. This difference was related to the high IgG content of the early milk (12·6 mg/ml v. 0·5 mg/ml for the mature milk) and stresses the importance of monitoring the IgG level of milk to assess the absence of colostrum.

High somatic cell count (SCC) affects milk quality and cheesemaking, resulting in a reduction in cheese yield and quality. In dairy cows, quarter milk samples with >200000 cells/ml are considered to have subclinical mastitis, while there is much uncertainty on the corresponding levels of SCC in buffalo milk. In this study 30 lactating water buffaloes were selected and SCC, differential somatic cell counts and several whey components were tested in quarter milk samples to assess the relationship between inflammation markers and milk quality. Overall 236 quarter milk samples were considered. To evaluate the relationship between cellular markers (SCC, polymorphonuclear leucocytes, PMN, and N-Acetyl-β-glucosaminidase, NAGase) and other milk components, three classes were defined (low, medium and high). Analysis of milk yield showed a significant reduction in the high class of each of the three markers chosen. Overall, the highest class was characterized by significant changes in milk composition and a lower milk quality. The presence of an inflammatory status of the udder was frequent after the first trimester of lactation and in buffaloes with two or more parturitions. This study showed that significant changes in milk components can be observed when SCC are >400000 cells/ml, PMN are >50% and NAGase is >100 units. These thresholds could be suggested as levels to define udder health status in buffalo cows.

The welfare effects of increased milk production associated with the use of recombinant bovine somatotropin (rBST) on dairy operations in the USA were examined for 1996. Results that derived from three different estimates of the milk-production response to rBST were evaluated and compared. One estimate, derived from a survey of dairy producers in Connecticut, led to economic-impact estimates that were not statistically significant. A second, derived from a national survey that concentrated on the health and management of dairy cattle, led to estimates that were unbelievably high. A third, derived from a national survey that concentrated on the economics of dairy producers, provided the most reasonable estimates of economic impacts. Results of economic analysis, using the latter results, indicated that if rBST had not caused milk production to increase, then the market price of milk would have been 2·2±1·5 cents/kg higher, and the total value of the milk produced would have risen from $23·0±0·6 billion to $24·1±1·0 billion. A welfare analysis demonstrated that the increased milk production (and the reduced market price) associated with the use of rBST in the USA caused the economic surplus of consumers to rise by $1·5±1·0 billion, while the economic surplus of dairy producers fell by $1·1±$0·8 billion. Increased milk production associated with rBST yielded a total gain to the US economy of $440±280 million. An analysis of annual percent changes in the number of dairy cows per operation, milk production per cow, total milk production, total number of dairy cows, and total number of dairy operations in the USA suggested that the dairy industry's long-term economic growth path was stable from 1989–2001 inclusive, and did not receive a shock resulting from the introduction of rBST.

The objective was to compare the efficacy of two experimental Staphylococcus aureus mastitis bacterins and a currently marketed five-isolate-based Staph. aureus bacterin (Lysigin™, Boehringer Ingelheim Vetmedica, Inc.) with unvaccinated controls. Forty-seven Holstein-Friesian heifers were randomly assigned to one of four groups such that Group 1 (n=11) received a three-isolate experimental bacterin, Group 2 (n=11) received a five-isolate experimental bacterin, Group 3 (n=14) received Lysigin, and Group 4 (n=11) served as unvaccinated controls. Vaccinations were administered twice 28 d apart in late gestation. All groups were challenged with a heterologous strain of Staph. aureus (ATCC 29740) on days 6, 7, and 8 of lactation. Mastitis score, somatic cell count (SCC), milk culture yield, and total daily milk yield data were collected before and after challenge. All 47 cattle developed a Staph. aureus IMI post-challenge with three animals in Group 1 and one animal in Group 3 clearing their Staph. aureus IMI by the end of the study. However, there was no evidence of a difference between vaccinates and control with regard to Staph. aureus clearance rates post-challenge (P[ges ]0·214). Cattle vaccinated with Lysigin had a lower mean duration of clinical mastitis and lower total mastitis score post-challenge than controls (P=0·045 and P=0·046, respectively). Overall, there was no evidence that any of the vaccinated groups had a lower mean SCC than control (P[ges ]0·148) for the tested study days. Likewise there was no evidence that vaccinates had greater milk yield than controls post-challenge (P=0·617). Hence, there was no evidence that the vaccines reliably prevented Staph. aureus IMI, but Lysigin showed benefit in reducing the clinical severity and duration of clinical disease post-challenge. Neither of the experimental bacterins appeared to perform better than Lysigin.

Microbial diversity of the raw milk for the production of Fior di Latte di Agerola and its changes during cheesemaking were studied. Viable counts showed that at the end of curd ripening, loads of lactic acid bacteria, both mesophilic and thermophilic rods and cocci, higher than those commonly evidenced in similar cheeses produced by using natural or commercial starters, were detected. Identification of 272 isolates, supported by molecular diagnostic aids, evidenced representative cultures of a high number of bacterial taxa of interest as participating in the process, although most of the isolates belonged to Lactococcus lactis and Lactobacillus helveticus species. RAPD-PCR and REA-PFGE biotyping were performed for the isolates of the above species and it was shown that most of the strains isolated from the raw milk occurred during the whole cheesemaking process, and an active role of these strains in the fermentation was supposed. The results offer further proof of the importance of the raw milk as source of technologically interesting strains of lactic acid bacteria capable of driving the fermentation of traditional cheeses.

The five exons and the 5′ and 3′-untranslated regions (5′-UTR and 3′-UTR) of the oGH gene were screened for mutations using PCR-single strand conformation polymorphism (PCR-SSCP) procedures in 523 Serra da Estrela ewes and were found to be highly polymorphic. The region extending across and between the GH2-N and GH2-Z copies was sequenced allowing the design of primers for the specific PCR amplification of each copy. These were cloned and sequenced in 20 animals representative of all SSCP patterns. The corresponding genotypes were established for each copy following nucleotide sequencing of SSCP alleles. Twenty-four polymorphic sites were found at the GH2-N (or GH1) and fourteen at the GH2-Z copies. Eight amino acid substitutions were predicted at the GH2-N and six at the GH2-Z copies. Milk yield adjusted to 150 lactation days was analysed for the genotype of each oGH gene copy taken separately or together (associated genotypes) by restricted maximum likelihood (REML) through a univariate best linear unbiased prediction (BLUP) animal model with repeated measures. Significant associations between genotypes and milk yield were observed. Within GH2-N genotypes there was a milk yield differential of 21·4±0·2 l/150 d between the most (N7) and the least (N5) productive ones. Within GH2-Z genotypes there was a differential of 21·6±0·2 l/150 d between the most (Z8) and the least (Z1) productive ones. The effect of associated GH2-N and GH2-Z genotypes revealed a differential of 39·6±0·3 l/150 d between the most (N1+Z7) and the least (N3+Z2) productive associated genotypes. The results show that GH2-N and GH2-Z genotypes significantly affect milk yield in Serra da Estrela ewes. Moreover, the apparent joint effect of GH2-N and GH2-Z genotype could improve milk yield in 25% as compared with the mean milk production of the analysed population.

A sensitive PCR method amplifying an internal fragment of the major tail protein gene was developed to detect Lactobacillus delbrueckii subsp. lactis lytic bacteriophages in undefined, thermophilic whey starters used in Italy for production of Grana and Provolone cheeses. PCR was applied to several lytic Lb. delbrueckii subsp. lactis bacteriophages, which were highly diverse according to restriction analysis and phage host range. PCR detected the presence of phages in two out of 11 cultures, when applied to whey starters for Grana Padano cheese sampled from different cheese plants. The presence of actively growing phages in infected cultures was confirmed by traditional test. The PCR method proved to be useful to screen for the presence of Lb. delbrueckii subsp. lactis phages in thermophilic whey starters.

Maintaining dry matter intake (DMI) during the last 2 weeks of gestation is difficult, and a failure to do so is reported to have negative implications on postcalving DMI (Grummer, 1995) and subsequent milk production (Bertics et al. 1992). DMI at the end of gestation, but excluding the last 1–2 d before parturition, can decrease by up to 30% (Lodge et al. 1975; Bertics et al. 1992; Dewhurst et al. 2000). The extent of the decline varies and may be influenced by a number of factors, such as level of feeding precalving (Lodge et al. 1975; Murphy, 1999; Dewhurst et al. 2000) or amount of non-structural carbohydrates in the precalving diet (Coppock et al. 1972; Forbes, 1995). Most studies were undertaken with cows receiving total mixed ration (TMR) or conserved forage and concentrates. However, this may have been one of the reasons for the witnessed depression. Increasing the proportion of rapidly fermentable carbohydrates or alternatively decreasing the fibre concentration in the precalving diet appears to exacerbate this decline (Coppock et al. 1972; Minor et al. 1998). Research suggests that ruminants may actively select against concentrates as plasma oestrogen concentrations increase, such as happens in the weeks preceding parturition (Forbes, 1995). When hay was offered ad libitum, increasing concentrations of oestrogen (up to 400 μg infused intravenously) had no effect. However, when a complete pelleted diet was offered, a 90-μg infusion caused a lower feed intake, followed by a significant increase after treatment was stopped. The lack of depression in precalving DMI reported by Coppock et al. (1972) and Vazquez-Anon et al. (1994) when forage was the principal component of the diet, supports this.

Milk fat globule membranes (MFGM) were isolated from the milk of mid-lactation Holstein cows. The purified MFGM were fractionated using 1-dimensional SDS gels. Tryptic peptides from gel slices were further fractionated on a micro-capillary high performance liquid chromatograph connected to a nanospray-tandem mass spectrometer. Analysis of the data resulted in 120 proteins being identified by two or more unique peptide sequences. Of these 120 proteins, 71% are membrane associated proteins with the remainder being cytoplasmic or secreted proteins. Only 15 of the proteins identified in the cow MFGM were the same as proteins identified in previous mouse or human MFGM proteomic studies. Thus, the bulk of the proteins identified are new for bovine MFGM proteomics. The proteins identified were associated with membrane/protein trafficking (23%), cell signalling (23%), unknown functions (21%), fat transport/metabolism (11%), transport (9%), protein synthesis/folding (7%), immune proteins (4%) and milk proteins (2%). The proteins associated with cell signalling or membrane/protein trafficking may provide insights into MFGM secretion mechanisms. The finding of CD14, toll like receptor (TLR2), and TLR4 on MFGM suggests a direct role for the mammary gland in detecting an infection.

Seventeen heterofermentative lactobacilli isolated from kefir grains were characterized by molecular methods. Bacterial isolates were identified by amplification of 16S rRNA gene and analysis by Amplified Ribosomal DNA Restriction Analysis (ARDRA), using the restriction enzymes Hae III, Dde I, and Hinf I. ARDRA analysis of lactobacilli isolates showed, for each enzyme used, a same banding pattern between the heterofermentative lactobacilli and the reference strains Lactobacillus kefir JCM 5818 and Lb. kefir ATCC 8007. Other reference lactobacilli and one homofermentative isolate showed differences in at least one of these patterns. The 16S-23S rRNA spacer region was also used to discriminate the bacterial isolates at the species level. The data obtained from the analysis of spacer region confirmed that sequencing of this genome region is a good tool for a reliable identification of members of Lb. kefir species. Genotyping of isolates was performed by Random Amplified Polymorphic DNA (RAPD-PCR) analysis using M13, Coc, ERIC-2 and 1254 primers. Patterns obtained allowed the differentiation of isolates in three groups. The three clusters showed by RAPD-PCR analysis could be correlated with at least three different strains of Lb. kefir species in the group of heterofermentative lactobacilli isolates obtained from Argentinian kefir grains.

Composite milk somatic cell counts (CMSCC) from four separate datasets, containing 3338, 350, 1408 and 1404 herds, were used. All herds were enrolled in the Norwegian Dairy Herd Recording System (NDHRS). The aim was to investigate associations between CMSCC at calving or prior to drying-off and future CMSCC in the remaining or subsequent lactation. CMSCC was determined using Fossomatic 5000® cell counters (Foss Electric, Hillerød, Denmark) according to IDF recommendations (International Dairy Federation, 1984) and a natural logarithmic transformation of the CMSCC data (lnCMSCC) was performed. Results obtained were arranged according to parity and lactation stage and regression models and general linear mixed models were applied, the latter model to account for clustering between herds. The best associations between CMSCC at calving or prior to drying-off and future CMSCC in the remaining or subsequent lactation were found by using at least two CMSCC test days after calving or prior to drying-off. The geometric mean of the second and third or the first three CMSCC test days explained 50% of the variation in future CMSCC in first parity cows. This information was accessible at 151 days in milk (DIM) in bimonthly tested herds, and at 87 DIM for monthly tested herds. There was not a large difference using two or three single consecutive weighted CMSCC test days compared with the geometric mean of two or three CMSCC test days. Our findings indicate the need of using at least two CMSCC test days and, if only one CMSCC test day is used, it should be obtained after 14 d post-calving or preferably after 30 DIM.

Oxytocin (OT) release and lactation performance in primiparous Syrian Shami cows were evaluated in response to two different machine milking regimes. Six cows were milked in the presence of the calves (PC) and subsequently suckled, whereas six cows were exclusively machine milked without the presence of their calves (WC) until day 91 post partum. Milk yield and milk constituents were determined weekly. The degree of udder evacuation was determined by the succeeding removal of residual milk. PC released OT during the milking process, whereas in WC no OT release was detected throughout the milking process. Consequently, the residual milk fraction was much lower in PC than in WC (11% v. 58%, P<0·05) and daily milk yield until day 91 post partum was higher in PC than in WC (12·6±0·3 v. 7·1±0·4 kg, P<0·05). In conclusion, Syrian Shami cattle are not suitable to be exclusively machine milked without the presence of their calves.

The objective was to utilize data from modern US dairy cattle to determine the effect of days dry on fat and protein yield, fat and protein percentages, days open, and somatic cell score in the subsequent lactation. Field data collected through the dairy herd improvement association from January 1997 to December 2003 and extracted from the Animal Improvement Programs Laboratory national database were used for analysis. Actual lactation records calculated from test-day yields using the test-interval method were used in this study. The model for analyses included herd-year of calving, year-state-month of calving, previous lactation record, age at calving, and days dry as a categorical variable. Fat and protein yield was maximized in the subsequent lactation with a 60-d dry period. Dry periods of 20 d or less resulted in substantial losses in fat and protein yield in the subsequent lactation. In contrast to yields, a short dry period was beneficial for fat and protein percentages. Short dry periods also resulted in fewer days open in the subsequent lactation; however, this was entirely due to the lower milk yield associated with shortened dry period. When adjusted for milk yield, short dry periods actually resulted in poorer fertility in the subsequent lactation. Long days dry improved somatic cell score in the subsequent lactation. Herds with mastitis problems should be cautious in shortening days dry because short dry periods led to higher cell scores in the subsequent lactation compared with 60-d dry.

In in vivo study on 24 weaned piglets (8 per group), the survival rates of human isolates Lactobacillus gasseri K7 and LF221 were quantified by selective enumeration on MRS agar with rifampicin, and the presence of both strains in intestinal mucosa was examined. Faeces from individual animals were analysed for the number (cfu/g) of coliforms, lactobacilli, clostridia and both of the two probiotic strains during 2-weeks probiotic application period (5×1010 cfu of individual strain/day) and 1 week after the probiotic treatment was ceased. Samples of duodenum, jejunum and ileum of sacrificed animals (5th or 20th day) were also examined microbiologically. A great variability in the microflora of faeces and mucosa was observed even between equally treated animals. The survival of both Lb. gasseri strains was established by their detection in the faeces (2·5×105 to 3·3×105 cfu of K7 strain/g faeces; 4·5×105 to 5×105 cfu of LF221 strain/g). In two animals, the LF221 or K7 viable cells were found in the faeces 6 d after ceasing probiotic application. In both animals from the group fed with Lb. gasseri K7 that were sacrificed 5 d after weaning, the presence of K7 strain was found either in the mucosa of duodenum (140 cfu/10 cm2) and jejunum (170 cfu/10 cm2) or in the ileum (1600 cfu/10 cm2). LF221 cells were detected in the ileal mucosa of one piglet (820 cfu/10 cm2). The results demonstrated the capability of both tested strains of in vivo adhesion to intestinal mucosa and of temporary colonisation of the piglets' intestine.

The incorporation of caseins and whey proteins into acid gels produced from unheated and heat treated skimmed milk was studied by confocal scanning laser microscopy (CSLM) using fluorescent labelled proteins. Bovine casein micelles were labelled using Alexa Fluor 594, while whey proteins were labelled using Alexa Fluor 488. Samples of the labelled protein solutions were introduced into aliquots of pasteurised skim milk, and skim milk heated to 90 °C for 2 min and 95 °C for 8 min. The milk was acidified at 40 °C to a final pH of 4·4 using 20 g glucono-delta-lactone/l (GDL). The formation of gels was observed with CSLM at two wavelengths (488 nm and 594 nm), and also by visual and rheological methods. In the control milk, as pH decreased distinct casein aggregates appeared, and as further pH reduction occurred, the whey proteins could be seen to coat the casein aggregates. With the heated milks, the gel structure was formed of continuous strands consisting of both casein and whey protein. The formation of the gel network was correlated with an increase in the elastic modulus for all three treatments, in relation to the severity of heat treatment. This model system allows the separate observation of the caseins and whey proteins, and the study of the interactions between the two protein fractions during the formation of the acid gel structure, on a real-time basis. The system could therefore be a valuable tool in the study of structure formation in yoghurt and other dairy protein systems.

Colostrum and milk samples from twelve Tunisian camels were analysed for concentration of immunoglobulin G (IgG), α-lactalbumin (α-la), serum albumin (CSA) and lactoferrin throughout the first 14 milkings post partum (7 days of lactation) using single radial immunodiffusion assay. Concentrations (mg/ml, means±SD) at first milking were IgG, 100·7±60·4; α-la, 2·2±0·7; CSA, 8·5±3·6 and lactoferrin, 1·2±0·3. Large variations were recorded for IgG and CSA concentrations (11·8–211·1 mg/ml and 2·9–13·8 mg/ml respectively) Concentrations of IgG and CSA dropped abruptly in the subsequent milkings while α-la concentration increased until milking 5 and then decreased slowly. Lactoferrin dropped only from milking 7. Mean IgG concentrations were 3·6 and 2·5 mg/ml at milking 9 and 13 respectively. However, IgG concentration did not differ significantly, at the 1% level, from milkings 11 to 14. The contribution of CSA to the increase in whey proteins in early milks was greater than that described in the bovine and caprine species.

Seven lactobacilli and a variety of microflora extracted from twenty five commercial cheeses were grown on unsupplemented acid goat whey and screened for their capacity to hydrolyse whey proteins [α-lactalbumin (α-la) and β-lactoglobulin (β-lg)] and to generate peptides. Fermentations were performed aerobically or anaerobically at 37 °C using crude or pre-heated whey (10 min at 65, 75 or 85 °C). Under aerobic conditions, growth of lactobacilli was poor and protein hydrolysis did not occur. Anaerobic conditions slightly increased lactobacilli growth but neither β-lg hydrolysis nor peptide generation were observed. More than 50% of α-la was digested into a truncated form of α-la (±12 kDa) in crude whey and whey pre-heated at 65 °C. Twenty-five microflora extracted from raw milk cheeses were screened for their proteolytic activities on acid goat whey under the conditions previously described. Eight of them were able to hydrolyse up to 50% of α-la mainly during aerobic growth on crude or pre-heated whey. The corresponding hydrolysates were enriched in peptides. The hydrolysate involving microflora extracted from Comté cheese after or at 18 months ripening was the only one to exhibit hydrolysis of both α-la and β-lg. Microbiological analysis showed that microorganisms originating from Comté cheese and capable of growth on unsupplemented whey consisted of Candida parapsilosis and Lactobacillus paracasei. Fermentation kinetic profiles suggested that peptides were released from α-la hydrolysis. The co-culture of both microorganisms was required for α-la hydrolysis that occurred concomitantly with the pH decrease. During whey fermentation, Cand. parapsilosis excrete at least one protease responsible for α-la hydrolysis, and Lb. paracasei is responsible for medium acidification that is required for protease activation.

An examination of the economic effects of epidemiologic risk factors for Johne's disease identified regional and herd size differences as having the greatest impact. Having dairy cows that were not born on the operation was the most important factor over which individual producers had the most immediate control. Economic consequences associated with using multiple-cow-maternity housing and multiple-preweaned-calf housing were not statistically significant. Economic welfare analysis was applied, and the GUM Workbench was used to analyse uncertainties in the estimates of the economic impacts.