Evaluation of Multi-Mixtures of Acids for the Sample Preparation of Organic Soil Amendments for Multi-Element Determination by ICP OES

A method for the simultaneous determination of aluminum (Al), arsenic (As), calcium (Ca), cadmium (Cd), chromium (Cr), copper (Cu), iron (Fe), magnesium (Mg), manganese (Mn), phosphorus (P), lead (Pb), sulfur (S), selenium (Se), vanadium (V), and zinc (Zn) in organic soil amendments using microwave-assisted acid digestion and inductively coupled plasma optical emission spectroscopy (ICP OES) is proposed. Concentrated or diluted acids mixtures (HNO₃, HF, HBF₄, and H₃BO₃) combined or not with H₂O₂ were systematically evaluated in order to achieve the best digestion procedure for masses of around 150 mg of samples. Principal component analysis (PCA) was applied in order to choose the best acid mixture for digestion (3 mL HNO₃ + 1 mL HBF₄ + 2 mL H₂O). The determined concentrations were in accordance with certified values of CRM 029 at the 95% confidence level, according to the Student-t test. This acid mixture was successfully applied for the digestion of four organic soil amendment samples (organic fertilizers, substrates, and soil conditioners) and element determination.     

Re-ensiling effects on inoculated sorghum: Intake,apparent digestibility,nitrogen balance,and animal feeding behaviour

This study aimed to determine whether using inoculants and re-ensiling in sorghum silages affect the intake and feeding behaviour, apparent digestibility, digestible energy, and nitrogen balance of sheep in maintenance. Half the sorghum was inoculated on the day of harvest, and the other half received the same inoculation volume but with water. Sorghum was ensiled in 100 experimental metal drum silos of 200 L covered with plastic: 50 silos with inoculants and 50 without them. After 56 days, 25 drums with inoculants and 25 without were exposed to air for 48 h. The other silos remained closed. The treatments were arranged in a 2 × 2 factorial scheme with inoculation and re-ensiling. The sheep received the silages after 211 days of ensiling and 155 days of re-ensiling to determine the intake, apparent digestibility, and feeding behaviour of animals. The experimental design was a simultaneous double 4 × 4 Latin square. The re-ensiling process increased silage pH, N-NH₃.TN⁻¹, butyric acid, and propionic acid. The inoculant increased propionic acid in ensiled and re-ensiled silages. Lactic acid concentration presented a statistical interaction with the inoculated and ensiled silage, 58% higher than the other treatment averages. The ADFap digestibility showed a complex interaction, in which the control ensiled and the inoculated and re-ensiled silages were about 35% lower than the inoculated ensiled silage. The Nbal:Nint ratio (g.g⁻¹) in the inoculated and re-ensiled silage had lower nitrogen retention than intake compared with the other treatments. These results indicate that exposing inoculated silages to air does not compromise their use in sheep feeding.     

The Marine-Derived Macrolactone Mandelalide A Is an Indirect Activator of AMPK

The mandelalides are complex macrolactone natural products with distinct macrocycle motifs and a bioactivity profile that is heavily influenced by compound glycosylation. Mandelalides A and B are direct inhibitors of mitochondrial ATP synthase (complex V) and therefore more toxic to mammalian cells with an oxidative metabolic phenotype. To provide further insight into the pharmacology of the mandelalides, we studied the AMP-activated protein kinase (AMPK) energy stress pathway and report that mandelalide A is an indirect activator of AMPK. Wild-type mouse embryonic fibroblasts (MEFs) and representative human non-small cell lung cancer (NSCLC) cells showed statistically significant increases in phospho-AMPK (Thr172) and phospho-ACC (Ser79) in response to mandelalide A. Mandelalide L, which also harbors an A-type macrocycle, induced similar increases in phospho-AMPK (Thr172) and phospho-ACC (Ser79) in U87-MG glioblastoma cells. In contrast, MEFs co-treated with an AMPK inhibitor (dorsomorphin), AMPKα-null MEFs, or NSCLC cells lacking liver kinase B1 (LKB1) lacked this activity. Mandelalide A was significantly more cytotoxic to AMPKα-null MEFs than wild-type cells, suggesting that AMPK activation serves as a protective response to mandelalide-induced depletion of cellular ATP. However, LKB1 status alone was not predictive of the antiproliferative effects of mandelalide A against NSCLC cells. When EGFR status was considered, erlotinib and mandelalide A showed strong cytotoxic synergy in combination against erlotinib-resistant 11-18 NSCLC cells but not against erlotinib-sensitive PC-9 cells. Finally, prolonged exposures rendered mandelalide A, a potent and efficacious cytotoxin, against a panel of human glioblastoma cell types regardless of the underlying metabolic phenotype of the cell. These results add biological relevance to the mandelalide series and provide the basis for their further pre-clinical evaluation as ATP synthase inhibitors and secondary activators of AMPK.     

Listeria monocytogenes and Salmonella spp. in raw milk produced in Brazil: occurrence and interference of indigenous microbiota in their isolation and development

This study aimed to verify the occurrence of Listeria monocytogenes and Salmonella spp. in raw milk produced in Brazil. On account of the poor microbiological quality of this product, possible interference from the indigenous microbiota in these pathogens was also evaluated. Two-hundred and ten raw milk samples were collected in four important milk-producing areas in Brazil, tested for L. monocytogenes and Salmonella spp. presence, and for enumeration of indicator microorganisms: mesophilic aerobes, total coliforms and Escherichia coli. The interference of the indigenous microbiota in the isolation procedures was also tested, as well the frequency of naturally occurring raw milk strains with antagonistic activity against both pathogens. The pathogens were not isolated in any raw milk sample, but poor microbiological quality was confirmed by the high levels of indicator microorganisms. When present at high levels, the indigenous microbiota generated an evident interference in the methodologies of L. monocytogenes and Salmonella spp. isolation, mainly when the pathogens appeared at low levels. Three-hundred and sixty raw milk strains were tested for antagonistic activity against both pathogens, and 91 (25.3%) showed inhibitory activity against L. monocytogenes and 33 (9.2%) against Salmonella spp. The majority of the antagonistic strains were identified as Lactic Acid Bacteria species, mainly Lactococcus lactis subsp. lactis and Enterococcus faecium, known by antimicrobial substance production.     

Comparative IgG recognition of tick extracts by sera of experimentally infested bovines

Enzyme-linked immunosorbent assay (ELISA) and Western blot were used to investigate the pattern of antibody responses of six bovines infested twelve times with Rhipicephalus (Boophilus) microplus (Canestrini, 1887) (Acari: Ixodidae) (six heavy infestations followed by six light infestations) against salivary gland, gut and larvae extracts. During heavy infestations, bovine IgG levels were shown to be higher, and a decrease in the number and weight of ticks that completed the parasitic cycle was observed. The pattern changed starting from the seventh infestation, showing a decrease in IgG levels. An initial increase followed by a significant decrease in the proportion of ticks that completed the parasitic cycle was also observed from the seventh infestation. The number of molecules recognized by Western blot was higher from sera collected following heavy infestations than after light infestations, although a great variation in the profiles detected could be seen when the bovines were compared. These results indicate that IgG responses to different tick antigens may not be generally associated with bovine resistance, and that infestation levels modulate the magnitude of humoral responses and possibly the immune mechanisms in the natural acquisition of tick resistance.     

Miniature Ponies: Similarities and Differences from Larger Breeds in Follicles and Hormones during the Estrous Cycle

Recent direct and indirect comparisons between Miniature ponies and larger breeds during the estrous cycle are reviewed. There were many similarities but profound differences in the Miniature ponies, including (1) greatly reduced follicle population within each of several follicle diameter classifications, (2) smaller number of growing follicles 10 mm or larger per ovulatory wave (1.5 ± 0.3 vs. 5.8 ± 0.9 in Miniature ponies vs. horses), (3) lower incidence of detectable diameter deviation, (4) ovulatory luteinizing hormone (LH) surge that involved three distinct phases, and (5) LH increase and decrease during the early luteal phase that followed the ovulatory surge. Diameter of the preovulatory follicle was smaller in Miniature ponies than in horses, but the difference was slight (38 vs. 44 mm; 14%) when compared with an extreme difference in median body weight (123 vs. 465 kg; 74%). The great reduction in follicle population with only a slight reduction in diameter of the preovulatory follicle in Miniature ponies compared with larger breeds is an important consideration during both experimental and clinical examinations.     

Nitrogen‐fertilizer forms affect the nitrogen‐use efficiency in fertigated citrus groves

The fertigated area of the Brazilian citrus industry has grown rapidly during recent years, and an efficient management of nitrogen (N) application at these sites is required for sustainable citrus production. Therefore, a field trial with Valencia orange trees [Citrus sinensis (L.) Osbeck] on Swingle citrumelo rootstock (Citrus paradise Macfad. x Poncirus trifoliata L. Raf.) was conducted for 8 years to evaluate the effects of N rates (80, 160, 240 and 320 kg ha^–1 y^–1) applied by fertigation, either as ammonium nitrate (AN) or calcium nitrate (CN), on soil solution dynamics, fruit yield, nutritional status, and N‐use efficiency (NUE) of trees. The maximum fruit yield was reached with 240 kg N ha^–1 for AN, whereas a linear response and greater fruit yield was observed for N supplied as CN. The NUE was reduced for both N forms with increasing N rates. However, the NUE for CN was 14 to 38% greater than the NUE for AN. The lower fruit yield and NUE for AN compared to CN‐treated trees was associated with the increased acidification of the soil solution with increased AN rates (pH ≤ 4.0). This limited nitrification resulted in a high ammonium (NH$ _4^+ $ ) concentration in the soil solution and a reduction in the net absorption of cations by the trees, particularly calcium (Ca). Due to the improved ion balance as well as the higher pH of the soil solution (pH ≥ 6.3) and diminished NH$ _4^+ $ availability, gains in both fruit yield and NUE in fertigated citrus groves in tropical soils can be obtained with the use of CN as a source of N.