Effect of processing method (natural, washed, honey, fermentation, maceration) on the availability of heavy metals in specialty coffee.

The aim of this study was to determine the effect of various methods of processing, such as natural, washed, honey, anaerobic fermentation, and carbonic maceration, on the contents of heavy metals in green and roasted specialty coffees from various countries of origin (Ethiopia, Kenya, Rwanda, Burundi, Guatemala, Nicaragua, and Peru). The heavy metals aluminium (Al), nickel (Ni), chromium (Cr), cadmium (Cd), copper (Cu), and lead (Pb) were identified by a multi-element technique using inductively coupled plasma mass spectrometry. Mercury (Hg) content was determined by atomic absorption spectrometry. The processing method affected the contents of Hg, Al, Ni, Cr, Cd, and Pb in the green and roasted coffees (p < 0.001). Hg content varied in the green coffees processed by fermentation methods vs natural or washed methods (i.e. Rwandan and Guatemalan coffees). Cd content was highest in Guatemalan green coffee processed using carbonic maceration (0.062 mg/kg). Pb content differed between the Ethiopian and Rwandan roasted coffees, with the highest content in the washed method (0.252 mg/kg). The correlations between the contents of Cu and Al, Ni and Cr, and Pb and Cr were significant for both the roasted and green beans. In conclusion, the method of processing can affect the contents of heavy metals in green and roasted specialty coffees. Monitoring heavy metals when processing coffee with new methods, even though further processing such as roasting can substantially reduce their content in some cases, is therefore important.

Chili pepper extracts, capsaicin, and dihydrocapsaicin as potential anticancer agents targeting topoisomerases.

DNA topoisomerases regulate conformational changes in DNA topology during normal cell growth, such as replication, transcription, recombination, and repair, and may be targeted for anticancer drugs. A DNA topology assay was used to investigate DNA-damaging/protective activities of extracts from Habanero Red (HR), Habanero Maya Red (HMR), Trinidad Moruga Scorpion (TMS), Jalapeno (J), Serrano pepper (SP), Habanero Red Savina (HRS), Bhut Jolokia (BJ), and Jamaica Rosso (JR) peppers, demonstrating their inhibitory effect on the relaxation of pBR by Topo I. DNA topoisomerase II (Topo II) is proven therapeutic target of anticancer drugs. Complete inhibition of Topo II was observed for samples TMS, HR, and HMR. Extracts J and SP had the lowest capsaicin and dihydrocapsaicin content compared to other peppers. HR, HMR, TMS, J, S, HRS, BJ, JR extracts showed the anticancer effect, examined by MTS and xCell assay on the in vitro culture of human colon carcinoma cell line HCT116.

Development and evaluation of a fish feed mixture containing the probiotic Lactiplantibacillus plantarum prepared using an innovative pellet coating method.

Introduction: Due to the intensification of fish farming and the associated spread of antimicrobial resistance among animals and humans, it is necessary to discover new alternatives in the therapy and prophylaxis of diseases. Probiotics appear to be promising candidates because of their ability to stimulate immune responses and suppress the growth of pathogens. Methods: The aim of this study was to prepare fish feed mixtures with various compositions and, based on their physical characteristics (sphericity, flow rate, density, hardness, friability, and loss on drying), choose the most suitable one for coating with the selected probiotic strain Lactobacillus plantarum R2 Biocenol™ CCM 8674 (new nom. Lactiplantibacillus plantarum). The probiotic strain was examined through sequence analysis for the presence of plantaricin- related genes. An invented coating technology based on a dry coating with colloidal silica followed by starch hydrogel containing L. plantarum was applied to pellets and tested for the viability of probiotics during an 11-month period at different temperatures (4°C and 22°C). The release kinetics of probiotics in artificial gastric juice and in water (pH = 2 and pH = 7) were also determined. Chemical and nutritional analyses were conducted for comparison of the quality of the control and coated pellets. Results and discussion: The results showed a gradual and sufficient release of probiotics for a 24-hour period, from 104 CFU at 10 mi up to 106 at the end of measurement in both environments. The number of living probiotic bacteria was stable during the whole storage period at 4°C (108), and no significant decrease in living probiotic bacteria was observed. Sanger sequencing revealed the presence of plantaricin A and plantaricin EF. Chemical analysis revealed an increase in multiple nutrients compared to the uncoated cores. These findings disclose that the invented coating method with a selected probiotic strain improved nutrient composition and did not worsen any of the physical characteristics of pellets. Applied probiotics are also gradually released into the environment and have a high survival rate when stored at 4°C for a long period of time. The outputs of this study confirm the potential of prepared and tested probiotic fish mixtures for future use in in vivo experiments and in fish farms for the prevention of infectious diseases.

Feeding-Regime-Dependent Intestinal Response of Rainbow Trout after Administration of a Novel Probiotic Feed.

Intensive fish farming is associated with a high level of stress, causing immunosuppression. Immunomodulators of natural origin, such as probiotics or phytoadditives, represent a promising alternative for increasing the immune function of fish. In this study, we tested the autochthonous trout probiotic strain L. plantarum R2 in a newly developed, low-cost application form ensuring the rapid revitalization of bacteria. We tested continuous and cyclic feeding regimes with regard to their effect on the intestinal immune response and microbiota of rainbow trout. We found that during the continuous application of probiotic feed, the immune system adapts to the immunomodulator and there is no substantial stimulation of the intestinal immune response. During the cyclic treatment, after a 3-week break in probiotic feeding and the reintroduction of probiotics, there was a significant stimulation of the gene expression of molecules associated with both cellular and humoral immunity (CD8, TGF-ß, IL8, TLR9), without affecting the gene expression for IL1 and TNF-α. We can conclude that, in aquaculture, this probiotic feed can be used with a continuous application, which does not cause excessive immunostimulation, or with a cyclic application, which provides the opportunity to stimulate the immunity of trout, for example, in periods of stress.

Effect of Concentration of Flaxseed (Linum usitatissimum) and Duration of Administration on Fatty Acid Profile, and Oxidative Stability of Pork Meat.

Flaxseed is a common ingredient used for livestock feed. The aim of this work was to study the effect of a diet supplemented with flaxseed at 5% and 10% concentrations in the intervals of 3 and 6 weeks prior slaughter on fatty acid profile and oxidative stability of pork meat. Meat samples were collected after slaughter from each animal (five groups, n = 6). Samples of the musculus longissimus dorsi (MLD) and the musculus gluteobiceps (MGB) were selected. Chemical composition, fatty acid profile and oxidative stability during the storage of meat under chilling conditions (4 °C, 7 days) was analyzed. The addition of flaxseed significantly affected the composition of fatty acid profile and the shelf life of the produced meat. The fat content was changed in the experimental groups with 10% flaxseed supplementation (10.84% in MGB and 9.56% MLD) versus the control group. Despite the different concentrations of flaxseed, the best EPA/AA ratio was observed in the experimental groups fed with flaxseed supplementation for 3 weeks. The worst oxidative stability of meat samples (p < 0.05) was recorded in the experimental groups with the addition of flaxseed for 6 weeks, which was related to higher PUFA content in samples of the experimental groups and higher susceptibility of PUFAs to lipid oxidation. The oxidative stability of meat in the experimental group fed 5% flaxseed supplementation for 3 weeks was not affected.

Surviving of production probiotic strains in a selected application form.

Research in probiotics for aquaculture is at an early stage of development and much work is still needed. Lactiplantibacilli belong to the microorganisms most frequently used to prepare the probiotics. The available information is inconclusive, since few experiments with sufficiently robust design have been conducted to permit critical evaluation. The development of probiotics applicable to commercial use in aquaculture is a multistep and multidisciplinary process requiring both empirical and fundamental research, full-scale trials, and an economic assessment of its use. The aim of the study was to prepare a probiotic aquafeed via excipients and subsequently to observe the survival of probiotic bacterial cells in the feed during the nine months storage period at a refrigerator (4 °C) or room temperature (22 °C). The strain Lactobacillus plantarum R2 Biocenol (CCM 8674) (according to the new taxonomy Lactiplantibacillus plantarum), potentially usable as a probiotic in aquaculture, was administered to prepare the aquafeed. Better survival of probiotic bacterial cells was recorded in a samples of pellets A (Aquatex 41 HMD) compared to the samples of probiotic pellets B (Inicio 918-2). Since oxidation of fatty acids in feed affects the nutritional quality of individual feed components, we assume that higher amounts of oil in feed B negatively affected the survival of probiotic bacterial cells. The highest numbers of viable probiotic bacteria cells were recorded at 4 °C storage of probiotic feed samples. The number of lactiplantibacilli dropped from 7.30 log10CFU . g-1 to 5.57 log10CFU . g-1 after the nine months storage period of feed samples A at 4 °C. Temperature is considered as a critical factor influencing probiotic viability and survival during storage period.

Effect of autochthonous lactobacilli on immunologically important molecules of rainbow trout after bacterial infection studied on intestinal primoculture.

Nowadays, the aquaculture industry is one of the fastest growing industries. Intensive aquaculture has a negative impact on fish health. Probiotic bacteria are often used due to beneficial effect to health of host, e.i. decrease of diseases outbreaks, immunomodulatory effect or better utilization of feed. The aim of this work was to study the influence of probiotic bacteria on the immune response of trout intestinal cells in primoculture infected with pathogenic bacteria. In the experiment, we tested the effect of pre-treatment of intestinal cells with an autochthonous strain of Lactobacillus plantarum R2 Biocenol™ (CCM 8674) following infection with the most serious salmonid pathogens - Aeromonas salmonicida subsp. salmonicida (CCM 1307) and Yersinia ruckeri (CCM 6093). Tested probiotic strain reduced inflammation after A. salmonicida infection through decreased expression of pro-inflammatory cytokines and increased expression of anti-inflammatory cytokines. In contrast, after infection with Y. ruckeri, which causes immunosuppression, the probiotic strain stimulated immunity by up-regulation of expression of proinflammatory cytokines and suppressed the expression of anti-inflammatory cytokines. These results are a prerequisite for the immunomodulatory potential of the strain, but its action must be confirmed in subsequent in vivo experiments.

Heavy-Metal Contents and the Impact of Roasting on Polyphenols, Caffeine, and Acrylamide in Specialty Coffee Beans.

The aim of this study was to determine the effect of roasting on the contents of polyphenols (PPH), acrylamide (AA), and caffeine (CAF) and to analyze heavy metals in specialty coffee beans from Colombia (COL) and Nicaragua (NIC). Samples of NIC were naturally processed and COL was fermented anaerobically. Green beans from COL (COL-GR) and NIC (NIC-GR) were roasted at two levels, light roasting (COL-LIGHT and NIC-LIGHT) and darker roasting (COL-DARK and NIC-DARK), at final temperatures of 210 °C (10 min) and 215 °C (12 min), respectively. Quantitative analyses of PPH identified caffeoylquinic acids (CQA), feruloylquinic acids, and dicaffeoylquinic acids. Isomer 5-CQA was present at the highest levels and reached 60.8 and 57.7% in COL-GR and NIC-GR, 23.4 and 29.3% in COL-LIGHT and NIC-LIGHT, and 18 and 24.2% in COL-DARK and NIC-DARK, respectively, of the total PPH. The total PPH contents were highest in COL-GR (59.76 mg/g dry matter, DM). Roasting affected the contents of PPH, CAF, and AA (p < 0.001, p < 0.011 and p < 0.001, respectively). Nickel and cadmium contents were significantly higher in the COL-GR than in the NIC-GR beans. Darker roasting decreased AA content, but light roasting maintained similar amounts of CAF and total PPH.

Effect of Fungal Solid-State Fermented Product in Broiler Chicken Nutrition on Quality and Safety of Produced Breast Meat.

The aim of this work was to analyse the effect of addition of 10% (w/w) fermented bioproduct into commercial broiler feed on fatty acid profile, lipid oxidative stability, and sensory properties of chicken breast meat. The fermented bioproduct was prepared by fermentation of cornmeal by filamentous fungi Umbelopsis isabellina CCF 2412 in solid-state fermentation (SSF) process and the final bioproduct was enriched with gamma-linolenic acid and beta-carotene. In the experiment, 80 pieces of 1-day-old chickens COBB 500 were used. Half of them (control group) were fed only with commercial feed. Chickens of the experimental group were fed with commercial feed, and, from the 11th day of age until the time of slaughter (39th day), 10% of commercial feed was replaced with fermented bioproduct. Application of fermented bioproduct into commercial feed mixture positively influenced profile of fatty acids in breast meat. The amount of gamma-linolenic, alpha-linolenic, and oleic acids in fat of breast muscles was increased and n-6/n-3 ratio was significantly decreased. Profile and content of PUFAs did not change after thermal treatment of meat. Oxidative stability of fat and sensory properties of the meat during the storage (4°C, 7 days) of meat were not affected by fermented bioproduct.

Species identification and antimicrobial resistance of coagulase-negative staphylococci isolated from the meat of sea fish.

UNLABELLED: Seventy-eight isolates of staphylococci obtained from the meat of Theragra chalcogramma, Scomber scombrus, and Clupea harengus were identified and tested in this study. 16S rDNA sequence specific for the genus Staphylococcus was detected in all isolates with the help of PCR method. All of 78 isolates were coagulase-negative, and DNAse activity was only confirmed in 4 of them. The following species of staphylococci were identified using MALDI-TOF mass spectrometry: S. warneri (52%), S. epidermidis (33%), S. haemolyticus (6.4%), S. pasteuri (3.8%), S. sciuri (1.2%), S. capitis (1.2%), and S. hominis (1.2%). Antimicrobial resistance to 7 antibiotics was determined in each isolate with the help of agar dilution method. In general, resistance against ampicillin was observed in majority of isolates (87%). On the contrary, the best sensitivity of CoNS was determined to gentamicin (96%). Only 1 S. warneri strain showed resistance to cefoxitin. Furthermore, 83% of staphylococcal isolates were simultaneously resistant to 2 or more antibiotics. PRACTICAL APPLICATION: This study confirmed the need of monitoring antimicrobial resistance in coagulase-negative staphylococci not only in the meat of slaughter animals but also in retail marine fish. The results showed that MALDI-TOF mass spectrometry is useful, accurate, and rapid method for species identification of food pathogens including Staphylococcus spp.

Effect of quantum therapy on pork quality.

In this study the impact of quantum therapy on meat quality of slaughtered pigs was investigated. For this purpose the pigs were treated with different doses of magnet-infrared-laser (MIL) radiation. Animals were divided into four groups according to radiation doses (4096, 512, and 64 Hz, and control without application), which were applied in the lumbar area of musculus longissimus dorsi (loin) at various time intervals prior to the slaughter (14 d, 24 h, and 1 h). Animals were slaughtered and the meat quality was evaluated by determining of pH value (1, 3, and 24 h post slaughter), drip loss, colour, and lactic acid and phosphoric acid amounts. MIL therapy can be used in various fields of veterinary medicine as are surgery and orthopaedics, internal medicine, dentistry, pulmonology, gastroenterology, gynaecology, urology, nephrology, and dermatology. The results achieved showed that MIL radiation used in a short period before slaughter (1 h) can cause a change in the meat quality, as reflected by the non-standard development of pH values, increases in drip loss, and changes of meat colour.

Post-mortem changes in the concentration of lactic acid, phosphates and pH in the muscles of wild rabbits (Oryctolagus cuniculus) according to the perimortal situation.

In this study changes in the concentrations of lactate, phosphates, and pH values of water extracts of muscles of transported and hunted rabbits during ripening were determined. Concentrations of lactate were higher in the muscles of hunted rabbits. The highest differences were obtained 24h after kill/hunt. Concentrations of lactate in the muscles of hunted rabbits were decreasing, while in the muscles of transported rabbits we observed it to increase in the 7th day and then decrease in the 14th day. Higher concentrations of phosphates were found in the muscles of transported wild rabbits. During the ripening process concentrations of phosphates were decreasing in muscles of both groups. Muscles of hunted rabbits had lower pH values during the whole ripening process. Our research showed that concentrations of lactate, phosphates and pH value post-mortem depended on the perimortal situations.

Determination of lipid oxidation level in broiler meat by liquid chromatography.

An assay was conducted for the determination of malondialdehyde (MDA) levels in broiler meat. The method involves extraction of tissues with trichloroacetic acid (TCA) and reaction of the TCA extract with 2,4-dinitrophenylhydrazine (DNPH). After separation of the MDA-DNPH complex using a solid-phase extraction C18 column, samples were eluted with 1 mL acetonitrile. Aliquots of 20 microL acetonitrile were analyzed by liquid chromatography on reversed-phase C18 column (3 microm) with UV detection. The products were eluted isocratically with the mobile phase containing acetonitrile-water-acetic acid (39 + 61 + 0.2, v/v/v). The retention time for MDA-DNPH was 6.5 min, and the detection limit was 3.5 microg/kg. Two extraction methods (cold and hot) were also used in the study. The results showed that hot extraction increased results about 55.8% and recovery from samples spiked with 116.6 microg/kg was lower (74.6%) in comparison with cold extraction (94.7%).