Technical note: Development and validation of a new method for the quantification of soluble and micellar calcium, magnesium, and potassium in milk

Milk mineral content is a key trait for its role in dairy processes such as cheese-making, its use as source of minerals for newborns, and for all traits involving salt-protein interactions. This study investigated a new method for measuring mineral partition between soluble and micellar fractions in bovine milk after rennet coagulation. A new whey dilution step was added to correct the quantification bias due to whey trapped in curd and excluded volume. Moreover, the proposed method allowed the quantification of the diffusible volume after milk coagulation. Milk mineral content and concentration in whey, and diluted whey were quantified by acid digestion and inductively coupled plasma optical emission spectrometry. The repeatability of the method for micellar Ca, Mg, and K was between 2.07 and 8.96%, whereas reproducibility ranged from 4.01 to 9.44%. Recovery of total milk minerals over 3 spiking levels ranged from 92 to 97%. The proposed method provided an accurate estimation of micellar and soluble minerals in milk, and curd diffusible volume

Genetic (co)variances between milk mineral concentration and chemical composition in lactating Holstein-Friesian dairy cows

Milk mineral concentration is important from both the perspective of processing milk into dairy products and its nutritive value for human consumption. Precise estimates of genetic parameters for milk mineral concentration are lacking because of the considerable resources required to collect vast phenotypes quantities. The milk concentration of calcium (Ca), potassium (K), magnesium (Mg), sodium (Na) and phosphorus (P) in the present study was quantified from mid-IR spectroscopy on 12 223 testday records from 1717 Holstein-Friesian cows. (Co)variance components were estimated using random regressions to model both the additive genetic and within-lactation permanent environmental variances of each trait. The coefficient of genetic variation averaged across days-in-milk (DIM) was 6.93%, 3.46%, 6.55%, 5.20% and 6.68% for Ca, K, Mg, Na and P concentration, respectively; heritability estimates varied across lactation from 0.31 ± 0.05 (5 DIM) to 0.67 ± 0.04 (181 DIM) for Ca, from 0.18 ± 0.03 (60 DIM) to 0.24 ± 0.05 (305 DIM) for K, from 0.08 ± 0.03 (15 DIM) to 0.37 ± 0.03 (223 DIM) for Mg, from 0.16 ± 0.03 (30 DIM) to 0.37 ± 0.04 (305 DIM) for Na and from 0.21 ± 0.04 (12 DIM) to 0.57 ± 0.04 (211 DIM) for P. Genetic correlations within the same trait across different DIM were almost unity between adjacent DIM but weakened as the time interval between pairwise compared DIM lengthened; genetic correlations were weaker than 0.80 only when comparing both peripheries of the lactation. The analysis of the geometry of the additive genetic covariance matrix revealed that almost 90% of the additive genetic variation was accounted by the intercept term of the covariance functions for each trait. Milk protein concentration and mineral concentration were, in general, positively genetically correlated with each other across DIM, whereas milk fat concentration was positively genetically correlated throughout the entire lactation with Ca, K and Mg; the genetic correlation with fat concentration changed from negative to positive with Na and P at 243 DIM and 50 DIM, respectively. Genetic correlations between somatic cell score and Na ranged from 0.38 ± 0.21 (5 DIM) to 0.79 ± 0.18 (305 DIM). Exploitable genetic variation existed for all milk minerals, although many national breeding objectives are probably contributing to an indirect positive response to selection in milk mineral concentration

Validation of a gold standard method for iodine quantification in raw and processed milk, and its variation in different dairy species

Adequate milk consumption significantly contributes to meeting the human iodine recommended daily intake, which ranges from 70 µg/d for infants to 200 µg/d for lactating women. The fulfilment of iodine recommended daily intake is fundamental to prevent serious clinical diseases such as cretinism in infants and goiter in adults. In the present study iodine content was measured in raw and processed commercial cow milk, as well as in raw buffalo, goat, sheep, and donkey milk. Iodine extraction was based on 0.6% (vol/vol) ammonia, whereas iodine detection and quantification were carried out through an inductively coupled plasma mass spectrometer analyzer. Among processed commercial cow milk, partially skimmed pasteurized milk had the greatest iodine content (359.42 µg/kg) and raw milk the lowest (166.92 µg/kg). With regard to the other dairy species, the greatest iodine content was found in raw goat milk (575.42 µg/kg), followed by raw buffalo (229.82 µg/kg), sheep (192.64 µg/kg), and donkey milk (7.06 µg/kg). Repeatability of milk iodine content, calculated as relative standard deviation of 5 measurements within a day or operator, ranged from 0.96 to 1.84% and 0.72 to 1.16%, respectively. The overall reproducibility of milk iodine content, calculated as relative standard deviation of 45 measurements across 3 d of analyses and 3 operators, was 4.01%. These results underline the precision of the proposed analytical method for the determination of iodine content in milk

Helix straminea Briganti, 1825 in Italy (Gastropoda: Pulmonata): taxonomic history, morphology, biology, distribution and phylogeny

The land snail taxon Helix straminea Briganti, 1825 has been reintroduced as a valid species in 2014. We provide here a comprehensive account of its taxonomy, distribution, anatomy, phylogeny and karyology in Italy. An overview of the historical views on the validity of the species is presented and faunistic data are reviewed and implemented with new records from Campania and Basilicata. A lectotype is fixed for H. straminea from the syntypes stored in the Muséum d’Histoire Naturelle of Genève, as well as for three other taxa (Helix straminiformis Bourguignat, 1876, Helix yleobia Bourguignat, 1883 and Helix straminea ssp. elongata Bourguignat, 1860). Genital system, radula and karyotype are described for the first time. Molecular analysis of two mitochondrial genes combining GenBank data and the new sequences presented in this paper showed no differentiation between the northern and southern Italian populations. The conservation status of the species and its possible threats are discussed

Invited review: Iodine level in dairy products—A feed-to-fork overview

The theme of iodine in the dairy sector is of particular interest due to the involvement and the interconnection of several stakeholders along the dairy food chain. Iodine plays a fundamental role in animal nutrition and physiology, and in cattle it is an essential micronutrient during lactation and for fetal development and the calf's growth. Its correct use in food supplementation is crucial to guarantee the animal's recommended daily requirement to avoid excess intake and long-term toxicity. Milk iodine is fundamental for public health, being one of the major sources of iodine in Mediterranean and Western diets. Public authorities and the scientific community have made great efforts to address how and to what extent different drivers may affect milk iodine concentration. The scientific literature concurs that the amount of iodine administered through animal feed and mineral supplements is the most important factor affecting its concentration in milk of most common dairy species. Additionally, farming practices related to milking (e.g., use of iodized teat sanitizers), herd management (e.g., pasture vs. confinement), and other environmental factors (e.g., seasonality) have been identified as sources of variation of milk iodine concentration. Overall, the aim of this review is to provide a multilevel overview on the mechanisms that contribute to the iodine concentration of milk and dairy products

Helix straminea Briganti, 1825 in Italy (Gastropoda: Pulmonata): taxonomic history, morphology, biology, distribution and phylogeny

The land snail taxon Helix straminea Briganti, 1825 has been reintroduced as a valid species in 2014. We provide here a comprehensive account of its taxonomy, distribution, anatomy, phylogeny and karyology in Italy. An overview of the historical views on the validity of the species is presented and faunistic data are reviewed and implemented with new records from Campania and Basilicata. A lectotype is fixed for H. straminea from the syntypes stored in the Muséum d'Histoire Naturelle of Genève, as well as for three other taxa (Helix straminiformis Bourguignat, 1876, Helix yleobia Bourguignat, 1883 and Helix straminea ssp. elongata Bourguignat, 1860). Genital system, radula and karyotype are described for the first time. Molecular analysis of two mitochondrial genes combining GenBank data and the new sequences presented in this paper showed no differentiation between the northern and southern Italian populations. The conservation status of the species and its possible threats are discussed

Mycobacterium tuberculosis Complex Mycobacteria as Amoeba-Resistant Organisms

International audienceBackground: Most environmental non-tuberculous mycobacteria have been demonstrated to invade amoebal trophozoites and cysts, but such relationships are largely unknown for members of the Mycobacterium tuberculosis complex. An environmental source has been proposed for the animal Mycobacterium bovis and the human Mycobacterium canettii.Methodology/Principal Findings: Using optic and electron microscopy and co-culture methods, we observed that 89±0.6% of M. canettii, 12.4±0.3% of M. tuberculosis, 11.7±2% of M. bovis and 11.2±0.5% of Mycobacterium avium control organisms were phagocytized by Acanthamoeba polyphaga, a ratio significantly higher for M. canettii (P = 0.03), correlating with the significantly larger size of M. canetti organisms (P = 0.035). The percentage of intraamoebal mycobacteria surviving into cytoplasmic vacuoles was 32±2% for M. canettii, 26±1% for M. tuberculosis, 28±2% for M. bovis and 36±2% for M. avium (P = 0.57). M. tuberculosis, M. bovis and M. avium mycobacteria were further entrapped within the double wall of <1% amoebal cysts, but no M. canettii organisms were observed in amoebal cysts. The number of intracystic mycobacteria was significantly (P = 10−6) higher for M. avium than for the M. tuberculosis complex, and sub-culturing intracystic mycobacteria yielded significantly more (P = 0.02) M. avium organisms (34×104 CFU/mL) than M. tuberculosis (42×101 CFU/mL) and M. bovis (35×101 CFU/mL) in the presence of a washing fluid free of mycobacteria. Mycobacteria survived in the cysts for up to 18 days and cysts protected M. tuberculosis organisms against mycobactericidal 5 mg/mL streptomycin and 2.5% glutaraldehyde.Conclusions/Significance: These data indicate that M. tuberculosis complex organisms are amoeba-resistant organisms, as previously demonstrated for non-tuberculous, environmental mycobacteria. Intercystic survival of tuberculous mycobacteria, except for M. canettii, protect them against biocides and could play a role in their life cycle

Uncommon Trimethoxylated Flavonol Obtained from Rubus rosaefolius

This study shows the evaluation the antiproliferative effect of the extract, fractions, and uncommon compounds isolated from R. rosaefolius leaves. The compounds were identified by conventional spectroscopic methods such as NMR-H1 and C13 and identified as 5,7-dihydroxy-6,8,4′-trimethoxyflavonol (1), 5-hydroxy-3,6,7,8,4′-pentamethoxyflavone (2), and tormentic acid (3). Both hexane and dichloromethane fractions showed selectivity for multidrug-resistant ovary cancer cell line (NCI-ADR/RES) with total growth inhibition values of 11.1 and 12.6 μg/ml, respectively. Compound 1 also showed selective activity against the same cell line (18.8 μg/ml); however, it was especially effective against glioma cells (2.8 μg/ml), suggesting that this compound may be involved with the in vitro antiproliferative action

Molecular Longitudinal Tracking of Mycobacterium abscessus spp. during Chronic Infection of the Human Lung

<div><p>The <i>Mycobacterium abscessus</i> complex is an emerging cause of chronic pulmonary infection in patients with underlying lung disease. The <i>M. abscessus</i> complex is regarded as an environmental pathogen but its molecular adaptation to the human lung during long-term infection is poorly understood. Here we carried out a longitudinal molecular epidemiological analysis of 178 <i>M. abscessus</i> spp. isolates obtained from 10 cystic fibrosis (CF) and 2 non CF patients over a 13 year period. Multi-locus sequence and molecular typing analysis revealed that 11 of 12 patients were persistently colonized with the same genotype during the course of the infection while replacement of a <i>M. abscessus sensu stricto</i> strain with a <i>Mycobacterium massiliense</i> strain was observed for a single patient. Of note, several patients including a pair of siblings were colonized with closely-related strains consistent with intra-familial transmission or a common infection reservoir. In general, a switch from smooth to rough colony morphology was observed during the course of long-term infection, which in some cases correlated with an increasing severity of clinical symptoms. To examine evolution during long-term infection of the CF lung we compared the genome sequences of 6 sequential isolates of <i>Mycobacterium bolletii</i> obtained from a single patient over an 11 year period, revealing a heterogeneous clonal infecting population with mutations in regulators controlling the expression of virulence factors and complex lipids. Taken together, these data provide new insights into the epidemiology of <i>M. abscessus</i> spp. during long-term infection of the CF lung, and the molecular transition from saprophytic organism to human pathogen.</p></div