Presence of pathogen DNA in milk harvested from quarters is associated to changes in cows' milk yield and composition.

BACKGROUND: Intramammary infection is the result of invasion and multiplication of microorganisms in the mammary gland and commonly leads to mastitis in dairy animals. Although much has been done to improve cows' udder health, mastitis remains a significant and costly health issue for dairy farmers, especially if subclinical. In this study, quarter milk samples from clinically healthy cows were harvested to detect pathogens via quantitative PCR (qPCR) and evaluate changes in individual milk traits according to the number of quarters infected and the type of microorganism(s). A commercial qPCR kit was used for detection of Mycoplasma bovis, Mycoplasma spp., Staphylococcus aureus, coagulase-negative staphylococci (CNS), Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Prototheca spp., Escherichia coli, Klebsiella spp., Enterococcus spp. and Lactococcus lactis ssp. lactis. Quarter and pooled milk information of 383 Holstein, 132 Simmental, 129 Rendena, and 112 Jersey cows in 9 Italian single-breed herds was available. RESULTS: Among the cows with pathogen(s) present in at least 1 quarter, CNS was the most commonly detected DNA, followed by Streptococcus uberis, Mycoplasma bovis, and Streptococcus agalactiae. Cows negative to qPCR were 206 and had the lowest milk somatic cell count. Viceversa, cows with DNA isolated in ≥ 3 quarters were those with the highest somatic cell count. Moreover, when major pathogens were isolated in ≥ 3 quarters, milk had the lowest casein index and lactose content. In animals with pathogen(s) DNA isolated, the extent with whom milk yield and major solids were impaired did not significantly differ between major and minor pathogens. CONCLUSIONS: The effect of the number of affected quarters on the pool milk quality traits was investigated in clinically healthy cows using a commercial kit. Results remark the important negative effect of subclinical udder inflammations on milk yield and quality, but more efforts should be made to investigate the presence of untargeted microorganisms, as they may be potentially dangerous for cows. For a smarter use of antimicrobials, analysis of milk via qPCR is advisable - especially in cows at dry off - to identify quarters at high risk of inflammation and thus apply a targeted/tailored treatment.

Pathogen Detection via Quantitative PCR in Milk of Healthy Cows Collected Using Different Sampling Protocols.

In this study we evaluated the prevalence of pathogens detected via quantitative PCR (qPCR) in milk from apparently healthy cows to identify the most common etiological agents present in Italian dairy farms. Milk samples were collected using a sterile protocol at quarter-level (3239 samples, 822 cows) and a conventional protocol at udder level as composite milk from the functional quarters of each cow (5464 samples, 5464 cows). The qPCR commercial kit detected Mycoplasma bovis, Mycoplasma spp., Staphylococcus aureus, coagulase-negative staphylococci (CNS), Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Prototheca spp., Escherichia coli, Klebsiella spp., Enterococcus spp. and Lactococcus lactis ssp. lactis as well as DNA from the penicillin resistance ß-lactamase gene from staphylococci. The prevalence of specific DNA was calculated based on its presence or absence in the samples, factoring in both the sampling protocols and herds. Regardless of the sampling protocol used, the most frequently detected pathogens were CNS (26.6% in sterile and 13.9% in conventional protocol) and Streptococcus uberis (9.6% and 16.5%, respectively). These results underscore the necessity for pathogen-specific interventions at the farm level to enhance the udder health of dairy cows via management recommendations.

ß-Casein A1 and A2: Effects of polymorphism on the cheese-making process.

Of late, "A2 milk" has gained prominence in the dairy sector due to its potential implications in human health. Consequently, the frequency of A2 homozygous animals has considerably increased in many countries. To elucidate the potential implications that beta casein (ß-CN) A1 and A2 may have on cheese-making traits, it is fundamental to investigate the relationships between the genetic polymorphisms and cheese-making traits at the dairy plant level. Thus, the aim of the present study was to evaluate the relevance of the ß-CN A1/A2 polymorphism on detailed protein profile and cheese-making process in bulk milk. Based on the ß-CN genotype of individual cows, 5 milk pools diverging for presence of the 2 ß-CN variants were obtained: (1) 100% A1; (2) 75% A1 and 25% A2; (3) 50% A1 and 50% A2; (4) 25% A1 and 75% A2; and (5) 100% A2. For each cheese-making day (n = 6), 25 L of milk (divided into 5 pools, 5 L each) were processed, for a total of 30 cheese-making processes. Cheese yield, curd nutrient recovery, whey composition, and cheese composition were assessed. For every cheese-making process, detailed milk protein fractions were determined through reversed-phase HPLC. Data were analyzed by fitting a mixed model, which included the fixed effects of the 5 different pools, the protein and fat content as a covariate, and the random effect of the cheese-making sessions. Results showed that the percentage of κ-CN significantly decreased up to 2% when the proportion of ß-CN A2 in the pool was ≥25%. An increase in the relative content of ß-CN A2 (≥50% of total milk processed) was also associated with a significantly lower cheese yield both 1 and 48 h after cheese production, whereas no effects were observed after 7 d of ripening. Concordantly, recovery of nutrients reflected a more efficient process when the inclusion of ß-CN A2 was ≤75%. Finally, no differences in the final cheese composition obtained by the different ß-CN pools were observed.

An observation of prescription behaviors and adherence to guidelines in patients with COPD: real world data from October 2012 to September 2014.

INTRODUCTION: GOLD guideline recommendations are currently the "gold standard" for the treatment of COPD patients. OBJECTIVES: The objective of this analysis was to evaluate compliance with GOLD guidelines in managing COPD patients' treatment by general practitioners (GPs) and pulmonologists. Since inhaled corticosteroid (ICS) use is defined as inappropriate in mild and moderate COPD patients, special attention was paid to ICS therapy use in these classes. METHODS: The study was based on the Italian GP database IMS Health Longitudinal Patient Database (IMS Health LPD) and on the Patient Analyzer specialist IMS Health database. The observed cohort included all patients with a diagnosis of COPD, aged 40 years or more, with at least one ATC R03 class prescription, visited by GPs and pulmonologists during four timeframes: October 2012 - March 2013 (cohort 1), April 2013 - September 2013 (cohort 2), October 2013 - March 2014 (cohort 3); April 2014 - September 2014 (cohort 4). Patients were classified into disease severity groups following 2008 GOLD guidelines, based on FEV1 value. RESULTS: Cohorts were quite similar in size (about two thousand patients per cohort). Pulmonologists visited more severe patients than GPs. About 50% of GPs' mild and moderate patients received treatments containing inhaled corticosteroids. Pulmonologists were more adherent to guidelines, with smaller percentages of mild patients treated with therapies containing ICS (ranging from 19.0% to 30.1%). An improvement in adherence was observed during the four time periods, with a decrease in the use of therapies containing ICS in mild and moderate patients. In absolute terms, it emerged that GPs more often prescribe ICS improperly to patients in the mild and moderate severity classes than pulmonologists. CONCLUSION: Real world data indicate that adherence to GOLD guidelines is only partially met by GPs in their general practice and shows higher prescription appropriateness by pulmonologists.

Survey on appropriateness of use of nimesulide in nine European countries.

BACKGROUND: Appropriateness of use is a key factor in safeguarding patient's health as well as a product's therapeutic properties. This paper presents the results of a survey conducted in nine European countries to verify the appropriateness of use of nimesulide in patients with inflammatory pain. METHODS: Computer-aided telephone interviews were administered to 1,277 nimesulide-prescribing general practitioners in Bulgaria, Czech Republic, Greece, Hungary, Italy, Poland, Portugal, Romania, and Slovakia, covering an estimated 31,719 patients. The interview questionnaire collected information on nimesulide prescriptions with respect to daily dose, treatment duration, and indication. RESULTS: In the majority of cases, prescriptions of nimesulide did not exceed the recommended daily dose of 200 mg (given as 100 mg twice a day), with a range from 161 mg (Greece) to 190 mg (Slovakia). An adherence to the 15-day treatment limit was observed in over 90% of cases. The average number of treatment days was always less than 15, with a range from 5.4 (Italy) to 13.6 (Czech Republic). Nimesulide was primarily used for the treatment of acute pain and short-term painful episodes in chronic conditions. The presence of gastrointestinal diseases/ulcers was the most frequent reason for not prescribing nimesulide. CONCLUSION: The results of this survey demonstrate that nimesulide is generally prescribed in compliance with the information reported in the summary of product characteristics (SmPC) with regard to daily dose and treatment duration, and suggest that it is mainly used for the management of episodes of acute pain in patients with a chronic disorder. These findings indicate the appropriateness of use of nimesulide in the European countries considered in this survey.

How do disease perception, treatment features, and dermatologist-patient relationship impact on patients assuming topical treatment? An Italian survey.

BACKGROUND: Psoriasis largely affects daily activities and social interactions and has a strong impact on patients' quality of life. Psoriatic patients have different attitudes toward their condition. Topical medications are essential for the treatment of psoriasis, but the majority of patients do not adhere to these therapies. OBJECTIVE: The history of treatment success or failure seems to influence patient attitude toward topical therapy. Therefore, it is important to understand the psychological, experiential, and motivational aspects that could be critical for treatment adherence, and to describe the different attitudes toward topical treatment. Furthermore, the physician-patient relationship and the willingness to trust the dermatologist may have a substantial role in encouraging or discouraging patients' attitudes toward topical therapy. METHODS: A survey was designed to collect aspects that could be relevant to understanding different patient attitudes toward psoriasis and its treatments. A total of 495 self-administered questionnaires compiled by psoriatic patients were analyzed from 20 Italian specialized hospital centers in order to provide a nationwide picture. RESULTS: Psoriatic patients have different perceptions and experiences in relation to their condition: half of them consider psoriasis as a disease, while the other half consider psoriasis as a disorder or a nuisance. Topical therapy is the most widely used treatment, even though it is not considered the most effective one and often perceived to be cosmetic. The main findings are: 1) inadequate patient education about this disease, 2) lack of information about topical treatment, and 3) lack of results within the expected time frame. Furthermore, physicians need to build a good relationship with psoriatic patients in order to motivate them, to trust in their care, and to adhere to treatment. CONCLUSION: This survey adds new and important details about daily life and well-being and the needs of psoriatic patients, providing suggestions for dermatologists to improve patients management.

Synthesis of indoles by intermolecular cyclization of unfunctionalized nitroarenes and alkynes, catalyzed by palladium-phenanthroline complexes.

Palladium-phenanthroline complexes efficiently catalyze the reaction of nitroarenes with arylalkynes and CO to give 3-arylindoles by an ortho-C-H functionalization of the nitroarene ring. Both electron-withdrawing and electron-donating substituents are tolerated on the nitroarene, except for bromide and activated chloride. Nitroarenes bearing electron-withdrawing substituents react faster, but the selectivity of the reaction depends on both polar and radical stabilization effects. Among those tested, only arylalkynes afforded indoles under the investigated conditions. The reaction mechanism was partly investigated. The kinetics is first order in nitroarene concentration and the rate-determining step of the cycle is the initial nitroarene reduction. No primary isotope effect is observed on either rate or selectivity, implying that the cyclization step is fast.