Evaluation of genetic polymorphism among Lactobacillus rhamnosus non-starter Parmigiano Reggiano cheese strains

Parmigiano Reggiano (PR) is an Italian cooked, long-ripened cheese made with unheated cow's milk and natural whey starter. The microflora is involved in the manufacturing of this cheese, arising from the natural whey starter, the raw milk and the environment. Molecular studies have shown that mesophilic non-starter lactic acid bacteria (NSLAB) are the dominant microflora present during the ripening of PR. In this study, a characterisation of Lactobacillus rhamnosus isolated from a single PR manufacturing and ripening process is reported, using a combination of genotypic fingerprinting techniques (RAPD-PCR and REP-PCR). The intraspecies heterogeneity evidenced for 66 strains is correlated to their abilities to adapt to specific environmental and technological conditions. The detection of biotypes that correlate with specific moments in cheese ripening or differential development throughout this process suggests that these strains may have specific roles closely linked to their peculiar technological properties.     

Segmentation of Parmigiano Reggiano dairies according to cheese-making technology and relationships with the aspect of the cheese curd surface at the moment of its extraction from the cheese vat

Parmigiano Reggiano cheese dairies develop specific cheese-making strategies to adapt the variable characteristics of raw, not standardized milk to the final goal of obtaining cheese consistent with the standard. Analyzing 1,175 cheese-making reports from 30 out of 383 dairies associated with the Parmigiano Reggiano Consortium in 2010 and 2011, 4 groups of Parmigiano Reggiano dairies using specific cheese-making technologies were discriminated by means of multiple linear discriminant analysis. Cheese makers manage cheese-making practices to obtain curd with different roughness properties, classified according to jargon words such as “rigata” and “giusta” or synonyms, because they believe that the roughness of the cheese curd surface immediately after the extraction from the vat is associated with different whey-draining properties and to the final outcome of the cheese. The aspect of the surfaces of the curds produced by the 4 groups of dairies was different according to the technology applied by each group. Cutting of the coagulum when it is still soft for a longer time and faster cooking of the cheese curd grains were associated with a less rough appearance of the surface of the curd, whereas under the opposite conditions, cutting the coagulum when it is firm for a shorter time, led to a curd with a rougher surface. These findings partially support the traditional feeling of Parmigiano Reggiano cheese makers, who consider the curd surface aspect one of the main drivers for their technological choices; to date, however, no data are provided about correlation between the aspect of the curd and the quality of the ripened cheese. If a sufficiently strong correlation could be demonstrated by the future development of the research, the operational effectiveness of Parmigiano Reggiano dairies will be able to largely benefit from the availability of sound and early process markers.     

Pyroglutamic acid in cheese: presence, origin, and correlation with ripening time of Grana Padano cheese

Pyroglutamic acid is present in many cheese varieties and particularly in high amounts (0.5 g/100 g of cheese) in extensively ripened Italian cheeses (Grana Padano and Parmigiano Reggiano) that are produced with thermophilic lactic acid bacteria as starters. The mechanism of pyroglutamic acid formation in cheese seems to be mostly enzymatic, as demonstrated by the presence of only L-pyroglutamic acid enantiomer. Thermophilic lactobacilli are involved in pyroglutamic acid production, as suggested by the low pyroglutamic acid content found in Bagos, a ripened Italian mountain cheese produced without addition of starter. Because milk pasteurization did not influence the pyroglutamic acid content in the ripened Grana Padano cheese, the formation of pyroglutamic acid mainly depends on the whey starter microflora rather than that of raw milk. Pyroglutamic acid concentration is linearly correlated (R2 = 0.94) with the age of Grana Padano cheese.     

Impact of area and sire by herd interaction on heritability estimates for somatic cell count in Italian Holstein Friesian cows.

The aim of the paper was to estimate variance components for somatic cell scores for Italian Holsteins using data from three different areas of the country. A total of 2,202,804 first-parity test-day records, collected from 1990 to 1997 in three areas of Italy (Mantova, Milano, and Parmigiano cheese area), were available for study. The areas differ in herd size, feeding systems and especially in milk use. A minimum standard of quality is also required by some specific methods of cheese production, as for example from the Parmigiano Reggiano cheese chain. These reasons, taken together, affect the attention given to the quality of milk production in herds, and, therefore, to the sanitation levels. A pedigree file was extracted from the national database of Holstein Friesian breed. For computational reasons, eight samples of the data were extracted per area. Variance components were estimated by sample using two different test-day repeatability models. The first model included fixed effects of herd-test date, days in milk (30-d intervals) and calving month, and random effects of permanent environment, additive genetic and residual error. Estimated heritabilities in the first model ranged from 0.06 to 0.09 and repeatabilities from 0.36 to 0.45. Only small differences were detected among areas. In the second model, a random sire x herd interaction effect was added. Including the sire x herd effect resulted in heritability estimates ranging between 0.05 and 0.08 and repeatabilities from 0.35 to 0.45. The analysis revealed that only a small fraction of the total variance (0.35 to 1.5%) could be explained by sire x herd interaction effect. Based on this research, it appears that parameter estimates for somatic cell count do not differ by region, and inclusion of a sire x herd interaction effect is unnecessary.     

Invited review: Microbial evolution in raw-milk,long-ripened cheeses produced using undefined natural whey starters

The robustness of the starter culture during cheese fermentation is enhanced by the presence of a rich consortium of microbes. Natural starters are consortia of microbes undoubtedly richer than selected starters. Among natural starters, natural whey starters (NWS) are the most common cultures currently used to produce different varieties of cheeses. Undefined NWS are typically used for Italian cooked, long-ripened, extra-hard, raw milk cheeses, such as Parmigiano Reggiano and Grana Padano. Together with raw milk microbiota, NWS are responsible for most cheese characteristics. The microbial ecology of these 2 cheese varieties is based on a complex interaction among starter lactic acid bacteria (SLAB) and nonstarter lactic acid bacteria (NSLAB), which are characterized by their different abilities to grow in a changing substrate. This review aims to summarize the latest findings on Parmigiano Reggiano and Grana Padano to better understand the dynamics of SLAB, which mainly arise from NWS, and NSLAB, which mainly arise from raw milk, and their possible role in determining the characteristics of these cheeses. The review is presented in 4 main sections. The first summarizes the main microbiological and chemical properties of the ripened cheese as determined by cheese-making process variables, as these variables may affect microbial growth. The second describes the microbiota of raw milk as affected by specific milk treatments, from milking to the filling of the cheese milk vat. The third describes the microbiota of NWS, and the fourth reviews the knowledge available on microbial dynamics from curd to ripened cheese. As the dynamics and functionality of complex undefined NWS is one of the most important areas of focus in current food microbiology research, this review may serve as a good starting point for implementing future studies on microbial diversity and functionality of undefined cheese starter cultures.     

Recovery and differentiation of long ripened cheese microflora through a new cheese-based cultural medium

A partial picture of the typical microflora of PDO Parmigiano Reggiano cheese was achieved by studying the cultivability of lactic acid bacteria associated with its manufacturing and ripening. A comprehensive sampling design allowed for the analysis of the cheese microflora during its production over 20 months of ripening. An innovative cheese agar medium (CAM) was prepared after testing 18 formulations all based on grated Parmigiano Reggiano ripened cheese. During cheese manufacturing and ripening, different samples were sampled and their microflora was recovered using CAM in comparison with other traditional media. Colonies which formed units from the different agar media tested were picked and isolated; the phylogenetic positions of 154 isolated strains were studied at level of species by 16S-rRNA gene sequencing. CAM seems to be able to recover the minority population coming from milk and whey starter, hardly estimable, during the first hours of production, on traditional media.     

Genome-wide association study for milk production and conformation traits in Canadian Alpine and Saanen dairy goats

Increasing the productivity of Canadian dairy goats is critical to the competitiveness of the sector; however, little is known about the underlying genetic architecture of economically important traits in these populations. Consequently, the objectives of this study were as follows: (1) to perform a single-step GWAS for milk production traits (milk, protein, and fat yields, and protein and fat percentages in first and later lactations) and conformation traits (body capacity, dairy character, feet and legs, fore udder, general appearance, rear udder, suspensory ligament, and teats) in the Canadian Alpine and Saanen breeds; and (2) to identify positional and functional candidate genes related to these traits. The data available for analysis included 305-d milk production records for 6,409 Alpine and 3,434 Saanen does in first lactation and 5,827 Alpine and 2,632 Saanen does in later lactations; as well as linear type conformation records for 5,158 Alpine and 2,342 Saanen does. Genotypes were available for 833 Alpine and 874 Saanen animals. Both single-breed and multiple-breed GWAS were performed using single-trait animal models. Positional and functional candidate genes were then identified in downstream analyses. The GWAS identified 189 unique SNP that were significant at the chromosomal level, corresponding to 271 unique positional candidate genes within 50 kb up- and downstream, across breeds and traits. This study provides evidence for the economic importance of several candidate genes (e.g., CSN1S1, CSN2, CSN1S2, CSN3, DGAT1, and ZNF16) in the Canadian Alpine and Saanen populations that have been previously reported in other dairy goat populations. Moreover, several novel positional and functional candidate genes (e.g., RPL8, DCK, and MOB1B) were also identified. Overall, the results of this study have provided greater insight into the genetic architecture of milk production and conformation traits in the Canadian Alpine and Saanen populations. Greater understanding of these traits will help to improve dairy goat breeding programs.     

The β-casein (CSN2) A2 allelic variant alters milk protein profile and slightly worsens coagulation properties in Holstein cows

Milk proteins genetic variants have long attracted interest as they are associated with important issues relating to milk composition and technological properties. An important debate has recently opened at an international level on the role of β-casein (β-CN) A1 and A2 polymorphisms, toward human health. For this reason, a lot of efforts has been put into the promotion of A2 milk by companies producing and selling A1-free milk, leading the farmers and breeders to switch toward A2 milk production without paying attention on the potential effect of the processability of milk into cheese. The aim of the present work was to evaluate the effects of β-CN, specifically the A1 and A2 allelic variants, on the detailed milk protein profile and cheese-making traits in individual milk samples of 1,133 Holstein Friesian cows. The protein fractions were measured with reversed-phase (RP)-HPLC (expressed in g/L and % N), and the cheese-making traits, namely milk coagulation properties, cheese yield, and curd nutrient recoveries assessed at the individual level, with a nano-scale cheese-making procedure. The β-CN (CSN2), κ-CN (CSN3), and β-lactoglobulin (LGB) genetic variants were first identified through RP-HPLC and then confirmed through genotyping. Estimates of the effects of protein genotypes were obtained using a mixed inheritance model that considered, besides the standard nuisance variables (i.e., days in milk, parity, and herd-date), the milk protein genes located on chromosome 6 (CSN2, CSN3) and on chromosome 11 (LGB), and the polygenic background of the animals. Milk protein genes (CSN2, CSN3, and LGB) explained an important part of the additive genetic variance in the traits evaluated. The β-CN A1A1 was associated with a significantly lower production of whey proteins, particularly of β-lactoglobulin (?8.2 and ?6.8% for g/L and % N, respectively) and α-lactalbumin (?4.7 and ?4.4% for g/L and % N, respectively), and a higher production of β-CN (6.8 and 6.1% for g/L and % N, respectively) with respect to the A2A2 genotype. Regarding milk cheese-making ability, the A2A2 genotype showed the worst performance compared with the other genotypes, particularly with respect to the BA1, with a higher rennet coagulation time (7.1 and 28.6% compared with A1A1 and BA1, respectively) and a lower curd firmness at 30 min. Changes in milk protein composition through an increase in the frequency of the A2 allele in the production process could lead to a worsening of the coagulation and curd firming traits.     

Headspace analysis of Italian and New Zealand parmesan cheeses

New Zealand is a leader in the global dairy industry. Milk powder is the principal export product, but there is also a prominent cheese manufacturing industry, catering more for the domestic market. The Selected Ion Flow Tube-Mass Spectrometric (SIFT-MS) technique was used to compare 4 New Zealand cheeses marketed as "parmesan" with 4 Italian Parmigiano Reggiano and Grana Padano cheeses. The cheese headspace was analyzed in real time without any sample preconcentration. Total of 38 volatile compounds in the cheese headspace were monitored with headspace concentrations varying between single digit parts per billion (ppb) to tens of parts per million (ppm). When the results were subjected to multivariate statistical analysis, a clear discrimination was found between the New Zealand "parmesan" and Italian cheeses based solely on the measured concentrations of these volatile compounds. If the volatile compounds used in the analyses were restricted to known odor-active compounds in Parmigiano Reggiano cheese, the ability to discriminate between the cheeses was maintained. The analyses also showed that it was possible to clearly differentiate between the different processing plants in individual countries. Important discriminatory volatiles in the samples tested were butanoic acid and phenylacetaldehyde for discriminating between Italian cheeses and ethyl butyrate, acetaldehyde and methylbutanals between New Zealand cheeses. We conclude that the New Zealand "parmesans" do not provide a good representation of the aroma of Italian "parmesans." PRACTICAL APPLICATION: SIFT-MS has been shown to clearly differentiate both country of origin and the manufacturer of "parmesan" cheeses made in Italy and New Zealand based on differences in volatile organic compounds. Thus this method will have benefit for use in the quality control of "parmesan" and other cheese varieties.     

Short communication: Confirmation of candidate causative variants on milk composition and cheesemaking properties in Montbéliarde cows

In a previous study, we identified candidate causative variants located in 24 functional candidate genes for milk protein and fatty acid composition in Montbéliarde, Normande, and Holstein cows. We designed these variants on the custom part of the EuroG10K BeadChip (Illumina Inc., San Diego, CA), which is routinely used for genomic selection analyses in French dairy cattle. To validate the effects of these candidate variants on milk composition and to estimate their effects on cheesemaking properties, a genome-wide association study was performed on milk protein, fatty acid and mineral composition, as well as on 9 cheesemaking traits (3 laboratory cheese yields, 5 coagulation traits, and milk pH). All the traits were predicted from midinfrared spectra in the Montbéliarde cow population of the Franche-Comté region. A total of 194 candidate variants located in 24 genes and 17 genomic regions were imputed on 19,862 cows with phenotypes and genotyped with either the BovineSNP50 (Illumina Inc.) or the EuroG10K BeadChip. We then tested the effect of each SNP in a mixed linear model including random polygenic effects estimated with a genomic relationship matrix. We confirm here the effects of candidate causative variants located in 17 functional candidate genes on both cheesemaking properties and milk composition traits. In each candidate gene, we identified the most plausible causative variant: 4 are missense in the ALPL, SLC26A4, CSN3, and SCD genes, 7 are located in 5′UTR (AGPAT6), 3′ untranslated region (GPT), or upstream (CSN1S1, CSN1S2, PAEP, DGAT1, and PICALM) regions, and 6 are located in introns of the SLC37A1, MGST1, CSN2, BRI3BP, FASN, and ANKH genes.     

Genome-wide association study for lactation persistency,female fertility,longevity, and lifetime profit index traits in Holstein dairy cattle

Female fertility in Holstein cattle can decline when intense genetic selection is placed on milk production. One approach to improving fertility is to identify the genomic regions and variants affecting fertility traits and then incorporate this knowledge into selection decisions. The objectives of this study were to identify or refine the positions of the genomic regions associated with lactation persistency, female fertility traits (age at first service, cow first service to conception, heifer and cow nonreturn rates), longevity traits (herd life, indirect herd life, and direct herd life), and lifetime profit index in the North American Holstein dairy cattle population. A genome-wide association study was performed for each trait, using a single SNP (single nucleotide polymorphism) regression mixed linear model and imputed high-density panel (777k) genotypes. No associations were identified for fertility traits. Several peak regions were detected for lifetime profit index, lactation persistency, and longevity. The results overlap with previous findings and identify some novel regions for lactation persistency. Previously proposed causative and candidate genes supported by this work include DGAT1, GRINA, and CPSF1, whereas new candidate genes are SLC2A4RG and THRB. Thus, the chromosomal regions identified in this study not only confirm several previous findings but also highlight new regions that may contribute to genetic variation in lactation persistency and longevity-associated traits in dairy cattle.     

Short communication: Association analysis of diacylglycerol acyltransferase (DGAT1) mutation on chromosome 14 for milk yield and composition traits,somatic cell score,and coagulation properties in Holstein bulls

The aim of the present study was to determine the allele frequencies of the diacylglycerol acyltransferase (DGAT1) K232A mutation in Italian Holstein bulls and to estimate the effect of the mutation on milk yield, composition, somatic cell score, and coagulation traits (rennet coagulation time and curd firmness). For this purpose, 349 Italian Holstein bulls were genotyped for the DGAT1 mutation on chromosome 14. Association analysis was performed by regressing the number of copies for the K allele on the deregressed estimated breeding value of the individual. Breeding values were calculated using field data routinely collected in Northeast Italy. The frequencies of the AA, KA, and KK genotypes were 59.6, 32.1, and 8.3%, respectively, and the minor allele frequency (K variant) was 24.7%. The K allele was significantly associated with greater fat yield and fat, protein, and casein percentages and with reduced protein:fat ratio. The association between the DGAT1 mutation and somatic cell score was not significant, whereas a favorable association between presence of the K allele and milk coagulation properties was found. Results from the present study confirmed the effect of the diallelic DGAT1 polymorphism K232A on milk production traits and, for the first time, provided evidence that this mutation also affects milk coagulation properties in the Italian Holstein breed.     

Genome-wide association study of milk fatty acid composition in Italian Simmental and Italian Holstein cows using single nucleotide polymorphism arrays

Bovine milk is important for human nutrition, but its fat content is often criticized as a risk factor in cardiovascular disease. Selective breeding programs could be used to alter the fatty acid (FA) composition of bovine milk to improve the healthiness of dairy products for human consumption. Here, we performed a genome-wide association study (GWAS) on bovine milk to identify genomic regions or specific genes associated with FA profile and to investigate genetic differences between the Italian Simmental (IS) and Italian Holstein (IH) breeds. To achieve this, we first characterized milk samples from 416 IS cows and 436 IH cows for their fat profile by gas chromatography. Subjects were genotyped with single nucleotide polymorphism array and a single-marker regression model for GWAS was performed. Our findings confirm previously reported quantitative trait loci strongly associated with bovine milk fat composition. More specifically, our GWAS results revealed significant signals on chromosomes Bos taurus autosome 19 and 26 for milk FA. Further analysis using a gene-centric approach and pathway meta-analysis identified not only some well-known genes underlying quantitative trait loci for milk FA components, such as FASN, SCD, and DGAT1, but also other significant candidate genes, including some with functional roles in pathways related to “Lipid metabolism.” Highlighted genes related to FA profile include ECI2, PCYT2, DCXR, G6PC3, PYCR1, and ALG12 in IS, and CYP17A1, ACO2, PI4K2A, GOT1, GPT, NT5C2, PDE6G, POLR3H, and COX15 in IH. Overall, the breed-specific association outcomes reflect differences in the genetic backgrounds of the IS and IH breeds and their selective breeding histories.     

Antibacterial activities of peptides from the water-soluble extracts of Italian cheese varieties

Water-soluble extracts of 9 Italian cheese varieties that differed mainly for type of cheese milk, starter, technology, and time of ripening were fractionated by reversed-phase fast protein liquid chromatography, and the antimicrobial activity of each fraction was first assayed toward Lactobacillus sakei A15 by well-diffusion assay. Active fractions were further analyzed by HPLC coupled to electrospray ionization-ion trap mass spectrometry, and peptide sequences were identified by comparison with a proteomic database. Parmigiano Reggiano, Fossa, and Gorgonzola water-soluble extracts did not show antibacterial peptides. Fractions of Pecorino Romano, Canestrato Pugliese, Crescenza, and Caprino del Piemonte contained a mixture of peptides with a high degree of homology. Pasta filata cheeses (Caciocavallo and Mozzarella) also had antibacterial peptides. Peptides showed high levels of homology with N-terminal, C-terminal, or whole fragments of well known antimicrobial or multifunctional peptides reported in the literature: α_S1-casokinin (e.g., sheep α_S1-casein (CN) f22-30 of Pecorino Romano and cow α_S1-CN f24-33 of Canestrato Pugliese); isracidin (e.g., sheep α_S1-CN f10-21 of Pecorino Romano); kappacin and casoplatelin (e.g., cow κ-CN f106-115 of Canestrato Pugliese and Crescenza); and β-casomorphin-11 (e.g., goat β-CN f60-68 of Caprino del Piemonte). As shown by the broth microdilution technique, most of the water-soluble fractions had a large spectrum of inhibition (minimal inhibitory concentration of 20 to 200 μg/mL) toward gram-positive and gram-negative bacterial species, including potentially pathogenic bacteria of clinical interest. Cheeses manufactured from different types of cheese milk (cow, sheep, and goat) have the potential to generate similar peptides with antimicrobial activity.     

Comparing local and commercial breeds on functional traits and profitability: the case of Reggiana dairy cattle

The objective of this study was to compare fertility, longevity, milkability, and profitability of cows from the Reggiana and Holstein breeds in northern Italy. Profitability was gauged for each breed, with consideration of economic incentive programs and alternative milk pricing scenarios. Calving to first service interval, days open, and calving interval were significantly shorter in Reggiana than in Holstein cows. Reggiana cows conceived approximately one estrus cycle before Holstein and had a calving interval 33 d shorter. Holstein cows released a significantly higher quantity of milk per unit of time (1.81 vs. 1.28 kg/min). Reggiana cows had longer expected total and productive lives than Holstein cows, by 5.8 and 10.0 mo, respectively. Replacement rate was 26% higher in the Holstein. Standard 305-d milk production was 5,360 and 7,870 kg in Reggiana and Holstein, respectively. Comparing breeds on annual milk and meat production, instead of standard 305-d milk yield, changed marginally the difference in annual profitability between the Reggiana and Holstein, from −€696 to −€679 per cow per year. Including feeding, milking, replacement, and insemination costs reduced the gap between breeds by 32%, from −€679, measured on annual milk and meat production, to −€460. These differences in profitability assumed a pricing scenario referring to milk sold to the dairy industry where protein and fat contents are valued but not the breed origin of milk. Incentive payments to farmers of endangered cattle compensated partially (22%) the lower income from Reggiana cows. When Reggiana milk production was sold as branded Parmigiano Reggiano cheese, Reggiana cows were more profitable than Holstein cows by €1,953 per cow per year.     

Quantification of aroma compounds in Parmigiano Reggiano cheese by a dynamic headspace gas chromatography-mass spectrometry technique and calculation of odor activity value

Potentially important aroma compounds in Parmigiano Reggiano cheese were quantified. Free fatty acids were isolated with ion-exchange chromatography and quantified by gas chromatography. Neutral aroma compounds were quantified with a purge-trap/gas chromatography-mass spectrometry with selective mass ion technique. Odor activity values were calculated based on sensory thresholds reported in literature. The calculated odor activity values suggest that 3-methylbutanal, 2-methylpropanal, 2-methylbutanal, dimethyl trisulfide, diacetyl, methional, phenylacetaldehyde, ethyl butanoate, ethyl hexanoate, ethyl octanoate, acetic, butanoic, hexanoic, and octanoic acids are the most important aroma contributors to Parmigiano Reggiano cheese.     

The occurrence of noncoagulating milk and the association of bovine milk coagulation properties with genetic variants of the caseins in 3 Scandinavian dairy breeds

Substantial variation in milk coagulation properties has been observed among dairy cows. Consequently, raw milk from individual cows and breeds exhibits distinct coagulation capacities that potentially affect the technological properties and milk processing into cheese. This variation is largely influenced by protein composition, which is in turn affected by underlying genetic polymorphisms in the major milk proteins. In this study, we conducted a large screening on 3 major Scandinavian breeds to resolve the variation in milk coagulation traits and the frequency of milk with impaired coagulation properties (noncoagulation). In total, individual coagulation properties were measured on morning milk collected from 1,299 Danish Holstein (DH), Danish Jersey (DJ), and Swedish Red (SR) cows. The 3 breeds demonstrated notable interbreed differences in coagulation properties, with DJ cows exhibiting superior coagulation compared with the other 2 breeds. In addition, milk samples from 2% of DH and 16% of SR cows were classified as noncoagulating. Furthermore, the cows were genotyped for major genetic variants in the α_S1- (CSN1S1), β- (CSN2), and κ-casein (CSN3) genes, revealing distinct differences in variant frequencies among breeds. Allele I of CSN2, which had not formerly been screened in such a high number of cows in these Scandinavian breeds, showed a frequency around 7% in DH and DJ, but was not detected in SR. Genetic polymorphisms were significantly associated with curd firming rate and rennet coagulation time. Thus, CSN1S1 C, CSN2 B, and CSN3 B positively affected milk coagulation, whereas CSN2 A², in particular, had a negative effect. In addition to the influence of individual casein genes, the effects of CSN1S1-CSN2-CSN3 composite genotypes were also examined, and revealed strong associations in all breeds, which more or less reflected the single gene results. Overall, milk coagulation is under the influence of additive genetic variation. Optimal milk for future cheese production can be ensured by monitoring the frequency of unfavorable variants and thus preventing an increase in the number of cows producing milk with impaired coagulation. Selective breeding for variants associated with superior milk coagulation can potentially increase raw milk quality and cheese yield in all 3 Scandinavian breeds.     

Effects of composite beta- and kappa-casein genotypes on milk coagulation, quality, and yield traits in Italian Holstein cows

The aim of the study was to estimate the effect of the composite CSN2 and CSN3 genotypes on milk coagulation, quality, and yield traits in Italian Holstein cows. A total of 1,042 multiparous Holstein cows reared on 34 commercial dairy herds were sampled once, concurrently with monthly herd milk recording. The data included the following traits: milk coagulation time; curd firmness; pH and titratable acidity; fat, protein, and casein contents; somatic cell score; and daily milk, fat, and protein yields. A single-trait animal model was assumed with fixed effects of herd, days in milk, parity, composite casein genotype of CSN2 and CSN3 (CSN2-CSN3), and random additive genetic effect of an animal. The composite genotype of CSN2-CSN3 showed a strong effect on both milk coagulation traits and milk and protein yields, but not on fat and protein contents and other milk quality traits. For coagulation time, the best CSN2-CSN3 genotypes were those with at least one B allele in both the CSN2 and CSN3 loci. The CSN3 locus was associated more strongly with milk coagulation traits, whereas the CSN2 locus was associated more with milk and protein yields. However, because of the tight linkage between the 2 loci, the composite genotypes, or haplotypes, are more appropriate than the single-locus genotypes if they were considered for use in selection.     

Short communication: Replication of genome-wide association studies for milk production traits in Chinese Holstein by an efficient rotated linear mixed model

Milk is regarded as an important nutrient for humans, and Chinese Holstein cows provide high-quality milk for billions of Chinese people. Therefore, detecting quantitative trait nucleotides (QTN) or candidate genes for milk production traits in Chinese Holstein is important. In this study, we performed genome-wide association studies (GWAS) in a Chinese Holstein population of 6,675 cows and 71,633 SNP using deregressed proofs (DRP) as phenotypes to replicate our previous study in a population of 1,815 cows and 39,163 SNP using estimated breeding values (EBV) as phenotypes. The associations between 3 milk production traits—milk yield (MY), fat percentage (FP), and protein percentage (PP)—and the SNP were determined by using an efficient rotated linear mixed model, which benefits from linear transformations of genomic estimated values and Eigen decomposition of the genomic relationship matrix algorithm. In total, we detected 94 SNP that were significantly associated with one or more milk production traits, including 7 SNP for MY, 76 for FP, and 36 for PP; 87% of these SNP were distributed across Bos taurus autosomes 14 and 20. In total, 83 SNP were found to be located within the reported quantitative trait loci (QTL) regions, and one novel segment (between 1.41 and 1.49 Mb) on chromosome 14 was significantly associated with FP, which could be an important candidate QTL region. In addition, the detected intervals were narrowed down from the reported regions harboring causal variants. The top significant SNP for the 3 traits was ARS-BFGL-NGS-4939, which is located within the DGAT1 gene. Five detected genes (CYHR1, FOXH1, OPLAH, PLEC, VPS28) have effects on all 3 traits. Our study provides a suite of QTN, candidate genes, and a novel QTL associated with milk production traits, and thus forms a solid basis for genomic selection and molecular breeding for milk production traits in Chinese Holstein.     

Effects of genetic type,stage of lactation,and ripening time on Caciocavallo cheese proteolysis

The objective of this experiment was to evaluate the effects of genetic type, stage of lactation, and ripening time on proteolysis in Caciocavallo cheese. One hundred twenty Caciocavallo cheeses made from the milk of 2 breeds, Italian Brown and Italian Holstein and characterized by different stages of lactation were obtained and ripened for 1, 30, 60, 90, and 150 d. Cheese proteolysis was investigated by ripening index (ratio of water-soluble N at pH 4.6 to total protein, %) and by the study of degradation of the protein fractions (α_S1-, β-, and para-κ-casein), which was determined by densitometric analysis of isoelectric focusing results. The statistical analysis showed a significant effect of the studied factors. Ripening index was higher in Italian Brown Caciocavallo cheese and in cheeses made with early lactation milk, whereas casein solubilization was greater in the first 2 mo of ripening. Isoelectric focusing analysis of cheese samples during ripening showed extensive hydrolysis of caseins. In particular, the protein fraction that underwent major degradation by proteolytic enzymes was α_S1-casein, followed by β-casein, whereas para-κ-casein was less degraded. Italian Brown cheese showed a lower residual quantity of β- and para-κ-casein, whereas Italian Holstein cheese showed a lower residual quantity of α_S1-casein. In addition, significant interactions of both first and second order were found on both ripening index and degradation of protein fractions. This study demonstrated that the analyzed factors influenced proteolysis of Caciocavallo cheese, which forms the basis of new knowledge that could lead to the production of a pasta filata cheese with specific characteristics.