Biological effects of a copper-based fungicide on the fruit fly, Drosophila melanogaster.

The increased consumption of pesticides can have a negative environmental impact by increasing the essential metals to toxic levels. Bordasul® is a commonly used fungicide in Brazil and it is composed of 20% Cu, 10% sulfur, and 3.0% calcium. The study of fungicides in vivo in non-target model organisms can predict their environmental impact more broadly. The Drosophila melanogaster is a unique model due to its ease of handling and maintenance. Here, the potential toxicity of Bordasul® was investigated by assessing the development, survival, and behavior of exposed flies. Exposure to Bordasul® impaired the development (p < 0.01) and caused a significant reduction in memory retention (p < 0.05) and locomotor ability (p < 0.001). Fungicides are needed to assure the world's food demand; however, Bordasul® was highly toxic to D. melanogaster. Therefore, Bordasul® may be potentially toxic to non-target invertebrates and new environmentally-safe biofertilizers have to be developed to preserve the biota.

Risk factors for oral mucositis in patients with solid tumors under treatment with cetuximab: a retrospective cross-sectional study.

BACKGROUND: This study retrospectively analyzed the risk factors for oral mucositis (OM) during cetuximab treatment. MATERIAL AND METHODS: We screened patients using cetuximab and retrospectively evaluated the presence of OM based on medical records. We collected information from 2 years of evaluations. Patient medical records were reviewed to obtain data on chemotherapy cycle and dose, sex, age, primary tumor, TNM stage, and head and neck radiotherapy (HNR) history. The X2 test and multinomial logistic regression were used for statistical analysis (SPSS 20.0, p < 0.05). RESULTS: Among 1831 patients, OM was showed in 750 in any grade (41%), during cetuximab treatment. Most patients were female (n=944, 51.6%), <70years-old (n=1149, 62.8%), had larynx cancer (n=789, 43.1%) in T4 (n=579, 47.7%), N0 (n=509, 52.6%) stages. Primary tumor surgery was performed in 1476 (80.6%) patients, radiotherapy in 606 (33.1%) patients and cetuximab protocols most used involved up to four cycles (n=1072, 58.5%) of <400mg (n=996, 54.4%) cetuximab doses. Female (OR [odds ratio] = 2.17, CI95% = 1.26-3.75), >70 years-old patients (OR = 16.02, CI95% = 11.99-21.41), with HHNR (OR = 1.84, 1.41-2.40), treated with >4 cycles (OR = 1.52, CI95% = 1.16-2.01) and high doses of cetuximab (OR = 3.80, CI95% = 2.52-5.71) are the greatest risk factors for OM. CONCLUSIONS: Since the clinical benefit of cetuximab in the treatment of older patients is limited and there is a high OM, especially in women with head and neck treated with radiotherapy, high doses and a high number of cetuximab cycles must be administered with caution.

Dysgeusia increases the risk for death and other side effects during antineoplastic systemic treatment for solid tumors: a cross-sectional study.

BACKGROUND: Chemotherapy (QT) is a systemic treatment using a combination of antineoplastic drugs, orally or intravenously, that inhibit tumor growth and fast-growing normal cells. Due to its nonspecificity, chemotherapy can cause a series of adverse effects, such as altered taste (dysgeusia), associated with malnutrition and, consequently, other adverse effects in the gastrointestinal tract and increased mortality risk. This study aimed to evaluate the influence of dysgeusia on the incidence of other adverse effects and overall survival during antineoplastic chemotherapy (chemotherapy). MATERIAL AND METHODS: An observational, retrospective, cross-sectional study was conducted using data from the Electronic Health Record system of the Cancer Institute of Ceará over two years. Before the CT session, the multi-professional team evaluated the patient for the presence and severity of adverse effects (AE), using scores from the CTCAE v5.0 scale. Dysgeusia scores were collected and associated with clinical pathological data, with other adverse effects (nausea, vomiting, diarrhea, oral mucositis, anorexia, constipation), and with overall survival. Chi-square and Mantel-Cox log-rank tests were used. RESULTS: Of 5744 patients evaluated, dysgeusia presented a frequency of 50.6%, being directly associated with female gender (p=0.001), overweight (p=0.022), high tumor stages (p=0.009), a combination of adjuvant and neoadjuvant (p=0.010) and four-year survival (p=0.030). Dysgeusia frequency was directly associated with diarrhea (p<0.001), anorexia (p<0.001), oral mucositis (p<0.001), nausea (p<0.001), constipation (p<0.001) and vomiting (p<0.001), and inversely associated with fatigue (p=0.035). CONCLUSIONS: Dysgeusia during CT increases the risk of other adverse effects and negatively impacts prognosis.

Pediatric postmortem CT angiography: validation of vascular access for PMCT angiography in stillbirths, babies and toddlers.

PURPOSE: The use of angiography in postmortem CT angiography (PMCTA) has several advantages. In adults, femoral vascular access is well established. Due to the small and specific anatomy in fetuses and infants, the technique has to be adapted, especially regarding the vascular access. The aim of this study was to evaluate vascular access for pediatric PMCTA (pedPMCTA). MATERIALS AND METHODS: Ten pedPMCTAs were performed in stillbirths, babies, and one toddler. A femoral approach by cannulation of the femoral artery and vein, an umbilical approach by cannulation of the umbilical vessels, and an intraosseous approach by an intraosseous needle were evaluated by handling and resulting imaging. RESULTS: The insertion of a cannula with a size of 18-20 G in the femoral vessels was possible in babies. An umbilical access with peripheral venous cannulas with a size of 14-20 G was feasible in stillbirths and newborns. An intraosseous access is advisable as equal alternative to umbilical and in cases where a femoral access is not possible. The most significant problem with the vascular access is the extravasation of contrast media, but this can be reduced significantly with practice. CONCLUSION: When performing pedPMCTA, an umbilical vascular access is recommended if an umbilical cord with open vessels is still present. Otherwise, a bone marrow access should be preferred in the presence of an arteriovenous shunt or if only the venous system needs to be shown. If that is not the case, the femoral access with the possibility to separate venous and arterial scan should be used.

The transCampus Metabolic Training Programme Explores the Link of SARS-CoV-2 Virus to Metabolic Disease.

Currently, we are experiencing a true pandemic of a communicable disease by the virus SARS-CoV-2 holding the whole world firmly in its grasp. Amazingly and unfortunately, this virus uses a metabolic and endocrine pathway via ACE2 to enter our cells causing damage and disease. Our international research training programme funded by the German Research Foundation has a clear mission to train the best students wherever they may come from to learn to tackle the enormous challenges of diabetes and its complications for our society. A modern training programme in diabetes and metabolism does not only involve a thorough understanding of classical physiology, biology and clinical diabetology but has to bring together an interdisciplinary team. With the arrival of the coronavirus pandemic, this prestigious and unique metabolic training programme is facing new challenges but also new opportunities. The consortium of the training programme has recognized early on the need for a guidance and for practical recommendations to cope with the COVID-19 pandemic for the community of patients with metabolic disease, obesity and diabetes. This involves the optimal management from surgical obesity programmes to medications and insulin replacement. We also established a global registry analyzing the dimension and role of metabolic disease including new onset diabetes potentially triggered by the virus. We have involved experts of infectious disease and virology to our faculty with this metabolic training programme to offer the full breadth and scope of expertise needed to meet these scientific challenges. We have all learned that this pandemic does not respect or heed any national borders and that we have to work together as a global community. We believe that this transCampus metabolic training programme provides a prime example how an international team of established experts in the field of metabolism can work together with students from all over the world to address a new pandemic.

Spatiotemporal trends of Chagas disease-related mortality in the Northeast of Brazil, 2007-2017.

Chagas disease (CD) is a neglected disease and endemic in Brazil. In the Brazilian Northeast Region, it affects millions of people. Therefore, it is necessary to identify the spatiotemporal trends of CD mortality in the Northeast of Brazil. This ecological study was designed, in which the unit of analysis was the municipality of the Brazilian northeast. The data source was the Information System of Mortality. It was calculated relative risk from socioeconomic characteristics. Mortality rates were smoothed by the Local Empirical Bayes method. Spatial dependency was analysed by the Global and Local Moran Index. Scan spatial statistics were also used. A total of 11 287 deaths by CD were notified in the study. An expressive parcel of this number was observed among 70-year-olds or more (n = 4381; 38.8%), no schooling (n = 4381; 38.8%), mixed-race (n = 4381; 62.3%), male (n = 6875; 60.9%). It was observed positive spatial autocorrelation, mostly in municipalities of the state of Bahia, Piauí (with high-high clusters), and Maranhão (with low-low clusters). The spatial scan statistics has presented a risk of mortality in 24 purely spatial clusters (P < 0.05). The study has identified the spatial pattern of CD mortality mostly in Bahia and Piauí, highlighting priority areas in planning and control strategies of the health services.

Increased expression of in situ IL-31RA and circulating CXCL8 and CCL2 in pemphigus herpetiformis suggests participation of the IL-31 family in the pathogenesis of the disease.

BACKGROUND: Pemphigus herpetiformis (PH) is a rare clinical subtype of pemphigus with the presence of urticarial plaques, severe pruritus, rare acantholysis and eosinophilic spongiosis. OBJECTIVES: The aim of this study was to investigate the influence of IL-31 and pro-inflammatory cytokines/chemokines in the pathogenesis of PH. METHODS: Twenty-five patients with PH and three groups: pemphigus foliaceus (PF = 14), pemphigus vulgaris (PV = 15) and healthy controls (HC = 20) were selected for this study. The groups were analysed by immunohistochemistry utilizing IL-31, IL-31RA, IL-4, IL-17 and TNF-α antibodies. Serum levels of IL-4, IL-13, TNF, CXCL8, CCL5 and CCL2 were evaluated by cytometric bead array. RESULTS: Analysis of IL-31 family of PH patients revealed the following findings: (i) Enhanced in situ expression of IL-31 in PH samples, compared to PF and to PV (epidermis); (ii) Cutaneous IL-31RA expression in PH samples was higher than in PF, PV and HC groups (epidermis and dermis); (iii) PF patients that evolved to PH showed significant increased IL-31RA epidermal expression during the PH phase. Profile of pro-inflammatory cytokines (IL-4, IL-17 and TNF-α) in PH patients' skin exhibited: (i) Enhanced IL-4 expression, when compared to patients with PF (epidermis and dermis) and with PV (epidermis); (ii) Augmented IL-17 expression than PF and PV patients (epidermis); (iii) Augmented expression of TNF-α when compared to PF at the epidermal level. Evaluation of circulating cytokines and chemokines showed higher levels of CXCL8 and CCL2 in PH sera compared to HC group. CONCLUSIONS: IL-31 and IL-31RA, cytokines related to pruritus, and pro-inflammatory chemokines (CXCL8 and CCL2) seem to exert a role in the pathogenesis of PH. These findings support future studies to clarify the role of IL-31 pathway as a potential therapeutic target for patients with PH.

Geochemical fractionation of hazardous elements in fresh and drilled weathered South African coal fly ashes.

The chemical reactions of dry-disposed ash dump, ingressed oxygen, carbon dioxide, and infiltrating rainwater affect mineralogical transformation, redistribution, and migration of chemical species. Composite samples of weathered coal fly ash taken at various depths and fresh coal fly ash were examined using organic petrographic, X-ray diffraction, X-ray fluorescence techniques, and successive extraction procedures. Results obtained show relative enrichment of glass, Al-Fe-oxides, calcite, and tridymite in the weathered CFA, but the fresh CFA is enriched in mullite, inertinite, maghemite, and ettringite. The enrichment of the weathered CFA in amorphous glass suggests higher reactivity when compared to fresh CFA. The evident depletion of soluble oxides in the weathered CFA is attributed to flushing of the soluble salts by percolating rainwater. Comparative enrichment of examined elements in water-soluble, exchangeable, reducible, and residual fractions of the weathered CFA is partly due to the slow release of adsorbed chemical species from the alumina-silicate matrix and diffusion from the deeper sections of the particles of coal fly ash. Sodium and potassium show enrichment in the oxidisable fraction of fresh CFA. The estimated mobility factor indicates mobility for Ca, Mg, Na, Se, Mo, and Sb and K, Sr, V, Cu, Cr, Se, and B in fresh and weathered CFAs, respectively.

Determination of macromineral requirements for preweaned dairy calves in tropical conditions.

International committees that have published nutrient requirements for dairy cattle have used data from mineral studies conducted in the 1920s to 1970s, and no study has reported data from animals less than 100 kg; therefore, there is a need to update mineral requirements for preweaned dairy calves. Thus, a meta-analysis was performed to estimate the mineral requirements of Ca, P, K, Mg, and Na for Holstein and Holstein × Gyr crossbred preweaned dairy calves using data from 5 studies developed at the Universidade Federal de Viçosa (Viçosa, MG, Brazil). A total of 210 calves were separated into 2 breeds: purebred Holstein calves (animals with a Holstein pedigree higher than 87.5%) and Holstein × Gyr crossbred calves (animals with a Holstein pedigree lower than 87.5%). The comparative slaughter technique was used to estimate animal body composition and empty body weight (EBW). Mineral requirements for maintenance were estimated by the regression between retained mineral and mineral intake, whereas mineral requirements for gain were obtained from the first derivative of the mineral content in the animal's body. In addition, breed effect was tested on the intercept and slope of the models. The effect of breed was not observed for all analyzed variables. Thus, net requirements for maintenance were 12.73, 11.81, 20.28, 3.50, and 6.37 mg/kg of EBW per day for Ca, P, K, Mg, and Na, respectively. Retention coefficients were 73.18, 65.20, 13.16, 29.55, and 24.28% for Ca, P, K, Mg, and Na, respectively. The following equations were determined to estimate net requirements for gain (NRG, g/d): NRG for Ca = 14.402 × EBW-0.139 × empty body gain (EBG); NRG for P = 5.849 × EBW-0.027 × EBG; NRG for K = 1.140 × EBW-0.048 × EBG; NRG for Mg = 0.603 × EBW-0.036 × EBG; and NRG for Na = 1.508 × EBW-0.045 × EBG. Due to the high variation between the data found in this study and in the available literature, we suggest that further studies should be conducted to evaluate the estimates of this study.

Energy and protein requirements for growth of Holstein × Gyr heifers.

There is little information regarding the nutritional requirements for dairy heifers, leading the majority of nutrient requirement systems to consider dairy heifers to be similar to beef heifers. Therefore, we evaluated the muscle protein metabolism and physical and chemical body composition of growing Holstein × Gyr heifers and estimated the energy and protein requirements. We performed a comparative slaughter experiment with 20 Holstein × Gyr heifers at an initial body weight of 218 ± 36.5 kg and an average age of 12 ± 1.0 months. Four heifers were designated as the reference group, and the 16 remaining heifers were fed ad libitum. The 16 heifers were distributed using a completely randomized design in a 2 × 2 factorial arrangement with two roughages (corn silage or sugarcane) and two concentrate levels (30 or 50%) for 112 days. Greater (p < 0.05) values for fractional rates of muscle protein synthesis, degradation and accretion were observed for heifers that were fed 50% concentrate. The following equations were obtained to estimate the net energy for gain (NEg ) and net protein for gain (NPg ): NEg (Mcal/day) = 0.0685 × EBW0.75  × EBWG1.095 and NPg (g/day) = 203.8 × EBWG - 14.80 × RE, respectively, in which EBW is the empty body weight, EBWG is the empty body weight gain and RE is the retained energy. We concluded that increased rates of protein turnover are achieved when a greater quality diet is provided. In the future, these results can be used to calculate the nutritional requirements for growth of Holstein × Gyr heifers after equation validation rather than using the recommendations provided by other systems, which use values developed from beef heifers, to determine the nutritional requirements of dairy cattle.

Antifungal activity of tyrosol and farnesol used in combination against Candida species in the planktonic state or forming biofilms.

AIM: This study examined the antifungal activity of the combination of tyrosol and farnesol against Candida albicans and Candida glabrata in the planktonic state or forming biofilms. METHODS AND RESULTS: The effect of drug association against Candida planktonic cells was assessed by the fractional inhibitory concentration index. Mono- and dual-species biofilms were developed during 24 h and then treated with the compounds for 3 days, with two daily treatments of 1 min each. After, the total biomass, metabolic activity and the number of cultivable cells were quantified. Planktonic cells of the two species showed a similar susceptibility to the drug combination, however, a synergistic effect was only verified for C. glabrata. Regarding biofilm susceptibility, significant reductions in C. glabrata biomass, metabolism of C. albicans and mixed biofilms, and cultivable cells of single biofilms were verified for the drug combination, indicating an additive effect. For all other experiments, the effects were classified as indifferent. CONCLUSION: The combined use of tyrosol and farnesol was advantageous for some of the analysed parameters against Candida species. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings may contribute to the development of oral care products containing tyrosol and farnesol to combat oral infections caused by Candida species.

Polarized Raman and Infrared Spectroscopy and ab Initio Calculation of Palmitic and Stearic Acids in the Bm and C Forms.

A complete experimental study on the vibrational properties of palmitic and stearic acids crystallized in the Bm and C forms, both belonging to the monoclinic system with the P21/a (C2h5) space group, through polarized Raman and infrared spectroscopy, is reported in this paper. Density functional theory calculations were also performed to assign the normal modes and to help in the interpretation of the experimental data. The different polarizations were compared and their influence on the spectral profiles, in both the lattice and the internal mode regions, was discussed. In general, the Raman and infrared spectra exhibit accentuated differences among the polymorphic forms, which are associated with the different molecular modifications, defined as gauche and all-trans conformations. Insights about interaction among different groups are also furnished.

Changes over time in the prevalence of asthma, rhinitis and atopic eczema in adolescents from Taubaté, São Paulo, Brazil (2005-2012): Relationship with living near a heavily travelled highway.

BACKGROUND: The prevalence of allergic diseases is increasing. We evaluated temporal trends in the prevalence of asthma, rhinitis and eczema in adolescents (13-14 years) living in Taubaté, SP, Brazil (2005-2012) and assessed the relationship between these prevalences and the residential proximity to Presidente Dutra Highway (PDH, a heavily travelled highway). METHODS: This cross-sectional study of adolescents (N=1039) from public and private schools was evaluated using the standard questionnaire of the International Study of Asthma and Allergies in Childhood (ISAAC) plus a question about their place of residence in relation to PDH. The data obtained were compared to the 2005 data using a chi-square test or Fisher's exact test. An analysis by groups consisting of two phases (two-step cluster) was used to evaluate the effect of living near PDH. RESULTS: There was a lifetime increase in the prevalence of active asthma (15.3% vs. 20.4%, p=0.005) and physician-diagnosed asthma (6.8% vs. 9.2%, p=0.06) and a decrease in the symptoms of active rhinitis (36.6% vs. 18.5%) between 2005 and 2012. A high frequency of asthma and rhinitis (18.1% vs. 23.2%, respectively) was observed among adolescents living close or very close to PDH; furthermore, 85.6% of the adolescents without symptoms of asthma or rhinitis lived far from PDH. CONCLUSION: An increase in the prevalence of asthma and a decrease in the prevalence of rhinitis were observed during the studied period. Living near PDH was associated with higher rates of asthma, rhinitis, and eczema.

Effects of day of gestation and feeding regimen in Holstein × Gyr cows: II. Maternal and fetal visceral organ mass.

This study investigated the influence of day of gestation (DG) and feeding regimens (FR) on maternal and fetal visceral organ mass in Holstein × Gyr cows. Forty-four pregnant multiparous Holstein × Gyr cows with an average initial body weight of 480±10.1 kg and an average initial age of 5±0.5 yr were allocated to 1 of 2 FR: ad libitum (AL; n=20) or maintenance level (ML; n=24). Maintenance level was considered to be 1.15% of body weight (dry matter basis) and met 100% of the energy requirements; AL provided 190% of the total net energy requirements. Cows were individually fed a corn silage and concentrate-based diet composed of 93% roughage and 7% concentrate (dry matter basis) as a total mixed ration twice daily. Pregnant cows were slaughtered at 4 DG: 139 (n=11), 199 (n=11), 241 (n=11), and 268 (n=11) d, which was followed by necropsy. Mass of heart, liver, and gastrointestinal tract was heavier in AL- than in ML-fed cows. Mammary gland mass was heavier in AL- than in ML-fed cows, and the heaviest mass was observed at 268 d of gestation. Feeding regimen did not influence fetal body weight in this study. The majority of the visceral organ masses were similar in fetuses from cows fed AL or ML. These data indicate that maternal feed restriction does not affect the development of most fetal organs or fetal development; however, some maternal organs are affected by the FR provided. Moreover, the negative effect on mammary gland mass caused by ML feeding will probably not affect the subsequent lactation because the crude protein concentration in the mammary gland increased with ML feeding. However, we suggest that the AL diet in pregnant dry cows should be provided with caution because the amount of fat in the mammary gland increased at 268 d of gestation.

Effects of day of gestation and feeding regimen in Holstein × Gyr cows: I. Apparent total-tract digestibility, nitrogen balance, and fat deposition.

This study investigated how feeding regimen (FR) alters apparent total-tract digestibility, performance, N balance, excretion of purine derivatives, and fat deposition in Holstein × Gyr cows at different days of gestation (DG). Forty-four pregnant multiparous Holstein × Gyr cows with an average initial body weight of 480±10.1 kg and an initial age of 5±0.5 yr old were allocated to 1 of 2 FR: ad libitum (AL; n=20) and maintenance level (ML; n=24). Maintenance level was considered to be 1.15% of body weight on a dry matter (DM) basis and met 100% of the energy requirements, whereas AL provided 190% of total net energy requirements. Data for hot and cold carcass dressing, fat deposition, average daily gain, empty body gain, and average daily gain without the gravid uterus were analyzed as a 4×2 factorial design. Intake, apparent total-tract digestibility, N balance, urinary concentration of urea, and purine derivatives data were analyzed as repeated measurements taken over the 28-d period (122, 150, 178, 206, 234, and 262 d of gestation). Cows were individually fed a corn silage-concentrate based diet composed of 93% roughage and 7% concentrate (DM basis) as a total mixed ration. Pregnant cows were slaughtered on 4 different DG: 139 (n=11), 199 (n=11), 241 (n=11), and 268 d (n=11). Overall, DM intake decreased as DG increased. This decrease observed in DM intake may be associated with the reduction in ruminal volume caused by the rapid increase in fetal size during late gestation. We observed an interaction for DM and organic matter apparent total-tract digestibility between FR and DG; at 150, 178, and 206 d of gestation, ML-fed cows had greater DM and organic matter apparent total-tract digestibility values than AL-fed cows. Rib fat thickness, mesentery, and kidney, pelvic, and heart fat were greater in AL-fed than in ML-fed cows at all DG, with the exception of rib fat thickness on d 139. Ad libitum-fed cows excreted more N in their feces and urine compared with ML-fed cows. Pregnant cows that were fed at maintenance had greater digestibility during some DG, excreted less N in feces and less N and urea in urine, and deposited less fat in the body. We therefore recommend ML (1.15% of body weight with 93% of roughage) as a FR for pregnant dry cows; however, during the last month of gestation, AL seems to be the most appropriate FR to avoid loss of body weight.

Effects of day of gestation and feeding regimen in Holstein × Gyr cows: III. Placental adaptations and placentome gene expression.

This study investigated the influence of day of gestation (DG) and feeding regimens (FR) on the expression of genes responsible for placenta development, nutrient transfer, and angiogenic factors in Holstein × Gyr cows. Forty pregnant multiparous Holstein × Gyr cows with an average initial body weight of 482±10.8kg and an initial age of 5±0.8 yr were allocated to 1 of 2 FR: ad libitum (AL; n=20) or maintenance level (ML; n=20). Maintenance level was considered to be 1.15% of body weight (dry matter basis) and met 100% of the net energy requirements and AL provided 190% of the total net energy requirements. Cows were slaughtered at 4 DG: 139, 199, 241, and 268d. After the cows were slaughtered, the placenta and uterus were separated and weighed. Caruncles and cotyledons were individually separated, counted, and weighed. Placenta expressed as kilograms and grams per kilogram of empty body weight (EBW) was heavier in ML- than in AL-fed cows at 268d of gestation. Placenta expressed as kilograms and grams per kilogram of EBW was the lightest at 139d of gestation, and the greatest mass was observed at 268d in ML-fed cows. However, in AL-fed cows, the heaviest placenta expressed as grams per kilogram of EBW was observed from 199d of gestation. Placentomes expressed as grams per kilogram of EBW were heavier in ML-fed cows during gestation, and the number of placentomes was greater in ML-fed cows at 268d of gestation. We observed that IGFR1 and IGFR2 were involved in placenta adaptations when ML was provided, as their expression in placentome cells was greater in ML-fed cows at 268d of gestation. The genes responsible for angiogenesis were also greater in ML-fed cows: VEGFA, GUCY1B3, HIFA, FGF2, and NOS3 were altered by FR and DG interaction and they were greater in ML-fed cows at 268d of gestation. In addition, VEGFB and ANGPT2 did not show interactions between FR and DG, but they were greater in ML-fed cows. Thus, we suggest that the placenta from an ML-fed cow develops adaptations to the reduced nutrient supply by altering its structure and gene expression, thereby developing mechanisms for potential increased nutrient transfer efficiency to the fetus.

Evaluation of methods of DNA extraction from Staphylococcus aureus in milk for use in real-time PCR.

The aim of this study was to evaluate the repeatability and performance of 4 methods of extracting DNA from Staphylococcus aureus (SAU) and the gene encoding bovine mitochondrial cytochrome B (BMCB) in milk samples from cows with subclinical mastitis for use in amplification by real-time polymerase chain reaction. Two milk samples were obtained from cows naturally infected with S. aureus and subjected to the following extraction methods: Qiagen DNA extraction kit; Axyprep DNA extraction kit; in silica column boil and in silica column method. After extraction in duplicate, eluates were subjected to purification and precipitation to determine purity (A260/A280 ratio) and concentration (µg/µL) by spectrophotometry and amplification by real-time polymerase chain reaction of target genes (SAU and BMCB). There was no effect of the DNA extraction method on DNA concentration and threshold cycle for BMCB and SAU. The purity ratio (A260/A280 ) was higher when using Qiagen DNA extraction (1.76 ± 0.136) compared to the other methods tested. Our results indicate that the DNA extraction kit from Qiagen produces samples of the highest purity ratio compared to other methods.

Exploring the potential of Burkholderia sacchari to produce polyhydroxyalkanoates.

AIM: Evaluation of the capability of Burkholderia sacchari to incorporate different monomers into polyhydroxyalkanoates (PHA). METHODS AND RESULTS: Thirty different carbon sources were evaluated as cosubstrates for B. sacchari growing on glucose with the intention to promote the incorporation of different monomers into the PHA produced by this species. With odd-numbered fatty acids, incorporation of the 3HV monomer was achieved, up to 65 mol% in the case of valerate. With 4-hydroxybutyrate, incorporation of 4HB was obtained, representing 9·1 mol%. With hexanoic acid, the production of P3HB-co-3HHx was achieved, containing up to 1·6 mol% of 3HHx. The molar fraction of 3HHx was found to be dependent on the ratio of glucose to hexanoic acid supplied. Metabolic flux analysis revealed a high efficiency of B. sacchari in converting carbon sources into P3HB-co-3HHx. Nevertheless, hexanoic acid was only poorly converted to 3HHx. CONCLUSIONS: Burkholderia sacchari is able to incorporate 3HV, 4HB and 3HHx in PHA containing mainly 3HB. The 3HHx content of P3HB-co-3HHx can be controlled by varying the glucose to hexanoic acid ratio. Burkholderia sacchari is highly efficient in converting carbon sources into PHA; however, only 2% of the hexanoic acid supplied could be converted to 3HHx. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report describing an approach to modulate the composition of P3HB-co-3HHx produced by bacteria using mixtures of carbohydrate and hexanoic acid as carbon source.

Detection and enumeration of Staphylococcus aureus from bovine milk samples by real-time polymerase chain reaction.

The aim of this study was to develop and evaluate a real-time quantitative PCR (qPCR)-based method to detect and quantify Staphylococcus aureus in bronopol-preserved milk samples from subclinical intramammary infections (IMI). Serial dilutions of milk artificially inoculated with Staph. aureus ATCC 29213 were used to establish a standard curve (cfu/mL) of the qPCR assay targeting the Staph. aureus thermonuclease-encoding gene nuc according to the strain plate count. The analytical sensitivity, specificity, and repeatability of the qPCR assay were determined. A total of 60 milk samples, collected from mammary quarters without abnormal appearance and with positive isolation of Staph. aureus, were submitted to both the qPCR protocol and Staph. aureus plate counting and results from both methods were compared. Staphylococcus aureus from bronopol-preserved, subclinical IMI milk samples were not accurately enumerated by qPCR compared with plate counting of the nonpreserved, raw milk sample. The detection limit of the qPCR protocol of inoculated Staph. aureus ATCC 29213 in bronopol-preserved milk samples was 1.04 × 10(1) cfu/mL. The qPCR protocol can be a high-throughput and rapid diagnostic assay to accurately detect Staph. aureus IMI from bronopol-preserved milk samples compared with a traditional culturing method. However, the proposed qPCR protocol is not accurate for counting of Staph. aureus in bronopol-preserved milk samples from naturally infected mammary glands.