Nadia Guzzo

Differential expression of living mammary epithelial cell subpopulations in milk during lactation in dairy cows

Epithelial cells are shed into milk during lactation, and although they generally reflect the cellular characteristics of terminally differentiated luminal cells, previously the detection of more primitive cells was described in human milk where a cell population of epithelial lineage was detected expressing markers typical of progenitor cells. In this investigation, we report the development of flow cytometry analysis to allow multiparametric assessment of mammary epithelial cells observed in milk. Cells collected from milk samples of 10 healthy dairy cows were directly analyzed for 6 different markers: CD45, CD49f, cytokeratin 14, cytokeratin 18, presence of nucleus, and cell viability. Milk samples were collected in 3 different periods of lactation: early lactation (EL=d 0-30), mid-lactation (ML=d 90-120), and late lactation (LL=210-250). Here we identify the differential expression of precursor or differentiated cell markers (or both) in mammary epithelial cells present in bovine milk. Myoepithelial cells, as indicated by cells staining positively for cytokeratin 14(+)/cytokeratin 18(-), were observed to increase from EL to LL with a high correlation with nuclear staining inferring potential proliferative activity. Furthermore, a significant increase in CD49f(+) and cytokeratin 14(+)/cytokeratin 18(+) positive cells was observed in LL. This assay is a sensitive approach for evaluating the variations in the frequency and features of living epithelial cells, whose reciprocal balance may be significant in understanding mammary gland cellular function throughout lactation. These observations suggest that mammary epithelial cell immunophenotypes could be investigated as biomarkers for mammary gland function in dairy cows.

Test day-milk yields variance component estimation using repeatability or random regression models in the Rendena breed

Abstract This study has aimed to compare Repeatability (RP-TDm) and Random-Regression Test Day models (RR-TDm) in genetic evaluations of milk (M), fat (F) and protein (P) yields in Rendena breed. Variance estimations for Milk (M), Fat (F) and Protein (P) were obtained on a sample of 43,842 TD belonging to 2,692 animals controlled over 15 years (1990-2005). RP-TDm estimates of h2 were of 0.21 for M and 0.17 for both F and P, whereas RR-TDM provided a trend of h2 ranging from 0.15-0.34 for M, 0.15-0.31 for F and 0.10-0.24 for P. Both RP-TDm and RR-TDm results agreed with literature, even though RR-TDm provided a pattern of h2 along the lactation different from other studies, with the lowest h2 at the beginning and at the end of lactation. PSB, MAD and -2Log L parameters revealed lower power of RP-TDm as compare with the RR-TDm.

In vivo performance of Italian Heavy Draft Horse weanlings fed two protein levels and slaughtered at two ages.

This study aimed at evaluating in vivo performance, growth parameters, intakes, dressing percentage, and blood parameters in Italian Heavy Draft Horse (IHDH) weanlings fed 2 CP levels up to the 2 typical ages of slaughter. Forty-one weanlings were grouped in 8 pens according to sex, age, and BW. After a transition period, animals were randomly assigned to 2 isoenergetic diets containing different CP levels: 10.6 and 11.2% CP in DM for low protein (LP) and 13.2 and 14.7% CP in DM for high protein (HP) diets in the first and second phase, respectively. About half of the animals (n = 22) were slaughtered when aged 13 mo (end of first phase); the remaining animals (n = 19) were slaughtered at 18 mo (end of second phase). Animals were weighed, measured for withers height, and scored in vivo for fleshiness and BCS at 3 wk intervals. Feed intake in each pen was measured weekly, and feed samples were collected every 2 mo. Blood samples from venous jugular were collected in both phases to analyze plasma protein, urea, glucose, bilirubin, hepatic enzymes, and mineral content. Growth parameters were estimated within phase by modeling BW as a function of age using fourth-degree Legendre polynomials. During the first phase, a different linear coefficient (P = 0.051) for the growth curve was observed between females fed a HP or a LP diet, while males showed differences only on quadratic and cubic Legendre coefficients. However, no significant differences were detected in ADG between the CP levels and sexes. In the second phase, Legendre coefficients were not different between treatments for the remaining weanlings, and once again no differences were found on ADG. The DM intake was influenced by diets in both periods, greater in the HP diet as compared with the LP diet (P < 0.001). No differences due to diet were observed for fleshiness or BCS scores at the end of each phase or in the dressing percentage at slaughter. As expected, plasma urea was greater (P < 0.001) in animals fed the HP diet but was within a normal range for healthy horses. In conclusion, a small dietary protein restriction (i.e., on average 3% of DM) did not change the in vivo performance of IHDH weanlings up to 13 or up to 18 mo of age.

Effects of total replacement of corn silage with sorghum silage on milk yield, composition, and quality

BackgroundIn the last years, difficulties occurring in corn cultivation (i.e., groundwater shortages, mycotoxin contamination) have been forcing dairy farmers to consider alternative silages. Some experiments conducted on lactating cows have proven that the total replacement of corn silage with sorghum silage did not reduce milk yield. However, this kind of substitution involves supplementing sorghum-based diets with grains, to compensate for the lower starch content of sorghum silage compared to corn silage. Change of silage type and inclusion of starch sources in the diet would influence rumen fermentations, with possible effects on milk composition (i.e., fatty acid profile) and coagulation properties. A worsening of milk coagulation properties would have a negative economic impact in Italy, where most of the milk produced is processed into cheese.This study was designed to compare milk composition and quality, with emphasis on fatty acid profile and coagulation properties, in dairy cows fed two diets based on corn or sorghum silage.ResultsThe sorghum diet reduced milk yield (P = 0.043) but not 4% fat corrected milk (P = 0.85). Feeding sorghum silage did not influence milk contents of protein (P = 0.07) and lactose (P = 0.65), and increased fat content (P = 0.024). No differences emerged for milk concentrations of saturated (P = 0.61) and monounsaturated fatty acids (P = 0.50), whereas polyunsaturated fatty acids were lower (P < 0.001) for the sorghum diet. Concentrations of n-6 (P < 0.001) and n-3 fatty acids (P = 0.017) were lower in milk of cows fed the sorghum diet. Milk coagulation properties did not differ between the two diets, except the “a30” (the curd firmness, expressed in mm, 30 min after rennet addition), that was lower (P = 0.042) for the sorghum diet.ConclusionsFeeding a forage sorghum silage, properly supplemented with corn meal, as total replacement of corn silage maintained milk composition and did not influence negatively milk coagulation properties, which have a great economic relevance for the Italian dairy industry. Thus, silages obtained from forage sorghums could have a potential as substitute of corn silages in dairy cow diets.

Genetic correlations among milk yield, morphology, performance test traits and somatic cells in dual-purpose Rendena breed

Selection in native local breeds needs great carefulness due to the small population size and the risk of inbreeding. Furthermore, most breeds are dual-purpose, and milk and beef attitudes are antagonistic. For preservation purposes functional traits need to be considered. Focusing on the small local Rendena cattle, this study aimed to analyse the genetic correlations among milk, beef and udder health traits and the response to selection predicted under different scenarios. The study considered milk, fat and protein yields (MY), factor scores for udder volume (UV), conformation (UC) and muscularity obtained from type traits scored on primiparous cows, and performance test traits (PT) measured on young bulls at test station: average daily gain, in vivo SEUROP fleshiness, in vivo dressing percentage. Somatic cell score (SCS) was considered as a functional trait, with a possibility of restricting its genetic gain to zero. The study considered 281 497 MY test-day data collected on 16 974 cows, and data from linear type evaluation on 11 992 primiparous cows for factor scores. The PT data were recorded on 1428 young bulls, and SCS obtained from cell counts at milk recording. Bi-trait restricted maximum likelihood animal model analyses were performed to assess genetic parameters. Heritability varied from 0.157 (fat) to 0.442 (dressing percentage). Udder volume and MY resulted positively genetically correlated (average correlation 0.427), whereas the low-negative genetic correlation between MY and UC (-0.141) suggested a negative impact of milk gain on udder form. Beef traits of factor muscularity and PT showed medium-high favourable genetic correlations (from 0.357 to 0.984), excluding a null correlation between daily gain and muscularity. The genetic correlation MY v. muscularity was unfavourable (-0.328 on average), whereas null correlations were found in MY v. PT, apart from fat v. dressing percentage (-0.151). Somatic cell score showed low unfavourable correlations with protein (0.111) and UV (0.092), and favourable correlations with UC (-0.193). Response to selection in different scenarios indicated a good balanced gain for milk and beef when standardized economic weights of 0.66 and 0.34 are given to the two attitudes, and SCS genetic gain is restricted. Current genetic trends (MY and PT increasing, but muscularity lessening) reflect a stronger selection for milk, suggesting a slight progressive change towards a milk conformation. Aiming to preserve the dual-purpose characteristics of a breed, proper breeding policies taking into account the genetic relationships among traits and including functional traits should be applied in local dual-purpose populations.

Genetic correlations between type and test-day milk yield in small dual-purpose cattle populations: The Aosta Red Pied breed as a case study

This study aimed at estimating the relationships between linear type traits and milk production in the dual-purpose Aosta Red Pied (ARP) cattle breed, by expressing type traits as factor scores with the same biological meaning of the individual traits. Factor analysis was applied to individual type traits for muscularity and udder of 32,275 first-parity ARP cows, obtaining 3 factor scores for individual muscularity (F1), udder side (F2), and udder conformation (F3). Data from 169,008 test-day records of milk, fat, and protein yield (kg), belonging to the first 3 lactations of 16,605 cows, were also analyzed. After obtaining genetic parameters for both morphological factors and milk production traits through a series of AIREML single-trait models, bivariate analyses were performed on a data set accounting for 201,283 records of 35,530 cows, to assess the phenotypic and genetic correlations among all factor scores and milk yield traits. The heritability estimates obtained proved to be moderate for both groups of traits, ranging from 0.132 (fat) to 0.314 (F1). Muscularity factor showed moderate and negative genetic correlations (ra) with udder size (-0.376) and udder conformation (0.214) factors. A low and negative ra was found between udder factors. Strong and positive ra were found among all the 3 milk production traits and F 0010 (ra≥0.597). Negative ra with milk traits were obtained for both F 0005 and F3, ranging from -0.417 to -0.221. Phenotypic correlations were lower than the genetic ones, and sometimes close to zero. The antagonism between milk production and meat attitude traits suggests that great attention should be paid in assigning proper weight to the traits, comprising functional traits such as udder conformation, included in selection indices for the dual-purpose breed. The ra obtained for factor scores are consistent with previous estimates for the corresponding individual type traits, and this confirms the possible use of factor analysis to improve type traits relevant to beef attitude.

Factor analysis for genetic evaluation of linear type traits in dual-purpose autochthonous breeds.

Factor analysis was applied to individual type traits (TT) scored in primiparous cows belonging to two dual purpose Italian breeds, Rendena (REN; 20 individual type traits evaluated on 11 399 first parity cows), and Aosta Red Pied (ARP; 22 individual type traits evaluated on 36 168 primiparous cows). Six common latent factors (F1 to F6; eigenvalues ⩾1) which explained 63% (REN) and 58% (ARP) of the total variance were obtained. F1 included TT mainly related to muscularity, and F2 to body size. The F3 and F4 accounted for udder size and conformation, respectively. F5 included rear legs and feet. Biological significance for F6 was not readily obtained. Moderate to low heritability were estimated through REML single-trait analysis from factor scores (from 0.22 to 0.52 in REN, and from 0.08 to 0.37 in ARP). The greatest heritability values were estimated for body size and muscularity (0.52 and 0.37 for body size; and 0.40 and 0.32 for muscularity in REN and ARP, respectively). As expected, rank correlations, obtained considering estimated breeding values derived from best linear unbiased prediction analysis on the individual TT and factor score, showed similar coefficients to those observed in the factor analysis following loading of TT within each latent factor. These results suggest the possibility to implement the factor analysis in the morphological evaluation, simplifying the information given by the type traits into new variables useful for the genetic improvement of dual purpose cattle.

Evolution of increased competitiveness in cows trades off with reduced milk yield, fertility and more masculine morphology

In some species females compete for food, foraging territories, mating, and nesting sites. Competing females can exhibit morphological, physiological, and behavioral adaptations typical of males, which are commonly considered as secondary sexual traits. Competition and the development of traits increasing competitiveness require much energy and may exert adverse effects on fecundity and survival. From an evolutionary perspective, positive selection for increased competitiveness would then result in evolution of reduced values for traits related to fitness such as fecundity and survival. There is recent evidence for such evolutionary trade‐offs involving male competition, but no study has considered competing females so far. Using data from competitions for dominance in cows (Bos taurus), we found negative genetic correlations between traits providing success in competition, that is, fighting ability and fitness traits related to milk production and with fertility (the inverse of parity‐conception interval). Fighting ability also showed low but positive genetic correlations with “masculine” morphological traits, and negative correlations with “feminine” traits. A genetic change in traits over time has occurred due to selection on competitiveness, corresponding to an evolutionary process of “masculinization” counteracting the official selection for milk yield. Similar evolutionary trade‐off between success in competition and fitness components may be present in various species experiencing female competition.

Evaluation of behaviour in stabled draught horse foals fed diets with two protein levels

The present work is aimed at evaluating the behaviour of Italian Heavy Draught Horse (IHDH) foals reared in semi-covered stables and fed two isoenergetic total mixed rations with different dietary protein levels (13.2% and 10.6% of CP on dry matter). The study was prompted by the restrictions for nitrate emissions in farms of the European Nitrate Directive. One suggested solution is to reduce dietary protein while maintaining normal performance and welfare, but there is a lack of literature in studies of horses. The behaviours of 20 foals of 437±60 kg of BW, aged 379±37 days and stabled in four pens by sex (S) and diet (D) were video recorded and analysed to build a suitable ethogram including 18 behaviours in six categories: ingestion, resting, maintenance, movement, social activities, other. The percentage of the daily time spent in each behavioural category and single behaviours was analysed via a single traits GLM including S, D and their interaction. Daily activity was consistent with existing literature: foals spent about 33% of the day in ingestion activities and 41% in resting, whereas social interactions constituted 8% of the time and individual maintenance <2%. Concerning diet, foals fed high protein spent more time in movement (19.62±0.73% of day v. 10.45±0.73% in low-protein (LP) foals; P⩽0.001), whereas the LP group increased resting (43.42±1.12% v. 38.02±1.12%; P⩽0.001). No stereotypies were found, and daily activity followed the typical values for draught breeds for foals in both dietary groups, a result that suggests the maintenance of well-being after dietary protein reduction. This result, together with the findings of a companion study showing no changes in growth performances of foals, showed that a reduction of CP in foal diet is reconcilable with the maintenance of performance and welfare.

Genetic parameters of different measures of somatic cell counts in the Rendena breed

The objective of this study was to consider different and alternative methods of using somatic cell count (SCC) data recorded according to the Italian official milk recording system, estimating its genetic parameters and the correlations with the yield traits (milk, fat, and protein) in the Rendena breed. The SCC traits defined for genetic evaluation were somatic cell score, log of the total daily SCC (LTSCC, i.e., SCC multiplied by daily milk yield) individually recorded in a day of official control, and 3 different thresholds (≥80,000, ≥150,000, and ≥400,000 cells/mL) for somatic cells. A total of 187,052 test-day monthly records of milk, fat, and protein yields and SCC belonging to 11,718 cows were used to estimate heritability and genetic correlations between SCC and yield traits via a bi-trait repeatability test-day model using a Bayesian approach. The heritability values estimated for the threshold traits ranged from 0.036 to 0.065, less than those observed for monthly somatic cell score and LTSCC traits that were equivalent to 0.088 and 0.103, respectively. Higher genetic correlations were estimated between LTSCC trait and all productive traits (0.379 for milk, 0.240 for fat, and 0.370 for protein). The other SCC traits considered have shown low or almost null genetic correlations with the productive traits (from 0.008 between fat yield and SCC ≥150,000 cells/mL to 0.234 between protein yield and SCC ≥400,000 cells/mL) and almost all estimates included zero in the 95% highest posterior density region interval. These results indicated that genetic selection for milk, fat, and protein production negatively affects the LTSCC content and SCC ≥400,000 cells/mL but does not negatively influence the other somatic cell and threshold SCC traits in the Rendena breed. However, the complete framework of genetic relationships of SCC with all traits under selection should be considered when deciding on the possible inclusion of SCC in the breeding program of this small cattle population.