ß-phenylethylamine as a novel nutrient treatment to reduce bacterial contamination due to Escherichia coli O157:H7 on beef meat

Bacterial infection by Escherichia coli O157:H7 through the consumption of beef meat or meat products is an ongoing problem, in part because bacteria develop resistances towards chemicals aimed at killing them. In an approach that uses bacterial nutrients to manipulate bacteria into behaviors or cellular phenotypes less harmful to humans, we screened a library of 95 carbon and 95 nitrogen sources for their effect on E. coli growth, cell division, and biofilm formation. In the initial screening experiment using the Phenotype MicroArray^TM technology from BioLog (Hayward, CA), we narrowed the 190 starting nutrients down to eight which were consecutively tested as supplements in liquid beef broth medium. Acetoacetic acid (AAA) and ß-phenylethylamine (PEA) performed best in this experiment. On beef meat pieces, PEA reduced the bacterial cell count by 90% after incubation of the PEA treated and E. coli contaminated meat pieces at 10 °C for one week.     

Evaluation of three compounds with insect juvenile hormone activity as grain protectants against insecticide-susceptible and resistant strains of Sitophilus species (Coleoptera: Curculionidae)

Three compunds with insect juvenile hormone activity have been tested as protectants for wheat grain against one insecticide-susceptible and two insecticide-resistant strains of Sitophilus granarius. Neither JH I nor methoprene gave complete control of S. granarius at 100 ppm whereas ethyl[2-(p-phenoxy)ethyl]carbamate was effective at only 5 ppm. There was no loss of biological activity of this last compound even after treated grain was stored at 25°C and 70% r.h. for 12 months, and it was also found to be highly active against susceptible strains of Sitophilus oryzae and Sitophilus zeamais. There was no evidence that two insecticide-resistant strains of S. granarius were cross-resistant to any of the compounds tested.     

Macromineral and trace element concentrations in milk from Finnish Ayrshire cows fed microalgae (Spirulina platensis) and rapeseed (Brassica napus)

Given the lack of research regarding the effect of microalgal supplementation in dairy cows on milk mineral concentrations, this study investigated the effect of feeding different protein supplements in dairy cow diets on milk, feces, and blood plasma mineral concentrations, associated milk and blood plasma transfer efficiencies, and apparent digestibility. Lactating Finnish Ayrshire cows (n = 8) were allocated at the start of the trial to 4 diets used in a replicated 4 × 4 Latin square design experiment: (1) control diet (CON), (2) a pelleted rapeseed supplement (RSS; 2,550 g/d), (3) a mixture of rapeseed and Spirulina platensis (RSAL; 1,280 g of RSS + 570 g of S. platensis per day), and (4) S. platensis (ALG; 1,130 g of S. platensis per day). In each of the 4 experimental periods, a 2-wk adaptation to the experimental diets was followed by a 7-d sampling and measurement period. Feed samples were composited per measurement period, milk, and feed samples (4 consecutive days; d 17–20), and blood plasma samples (d 21) were composited for each cow period (n = 32). Data were statistically analyzed using a linear mixed effects model with diet, period within square, square and their interaction as fixed factors, and cow within square as a random factor. Cows fed ALG were not significantly different in their milk or blood plasma mineral concentrations compared with CON, although feeding ALG increased fecal concentrations of macrominerals (Ca and Mg) and trace elements (Co, Cu, Fe, I, Mn, and Zn), and reduced their apparent digestibility, compared with CON. When compared with CON and ALG, milk from cows fed RSAL and RSS had lower milk I concentrations (?69.6 and ?102.7 μg/kg of milk, respectively), but total plasma I concentrations were not affected significantly. Feeding S. platensis to dairy cows did not affect mineral concentrations in cows' blood or milk, but care should be taken when rapeseed is fed to avoid reducing milk I concentrations which may in turn reduce consumers' I intake from milk and dairy products.     

Effectiveness of spinosad and spinetoram against three Sitophilus species: Influence of wheat endosperm vitreousness

The influence of wheat endosperm vitreousness on the effectiveness of spinosad and spinetoram against lab populations of Sitophilus granarius, Sitophilus oryzae (from Serbia) and Sitophilus zemais (from Serbia and Slovenia) were evaluated in a laboratory conditions. Characteristics of two test wheat varieties were determined, i.e. a variety with high (HVWG) and another with low (LVWG) endosperm vitreousness. Spinosad and spinetoram were applied at the doses of 0.5, 1 and 2 mg/kg (ppm) to both wheat varieties. Mortality was assessed after 7, 14 and 21 days, while impact on progeny production/reduction PR (%) was assessed after 8 weeks. All doses of spinosad applied to the HVWG variety and 1–2 mg applied to the LVWG variety achieved high mortality (93–100%) of S. granarius after 14 days, while high mortality of the other test species (97–100%) was achieved by 1–2 mg doses on the HVWG and 2 mg on LVWG varieties. After 14 days, all doses of spinetoram achieved high mortality (96–100%) of S. granarius on both wheat varieties, while high mortality of S. oryzae (97–100%) and both populations of S. zeamais (93–100%) was achieved using 1–2 mg doses on the HVWG and 2 mg dose on the LVWG variety. The highest S. granarius PR (>90%) was found in both wheat varieties treated with 2 mg/kg of spinosad or spinetoram, while the highest PR of S. zeamais was observed in HVWG treated with 2 mg/kg of spinosad and 1–2 mg/kg of spinetoram, and in LVWG treated with 2 mg/kg of spinetoram. The greatest PR (91–94%) of S. oryzae in both wheat varieties was observed only in grain treated with 2 mg/kg spinetoram. In general, endosperm vitreousness was found to influence the effectiveness of spinosyns to all Sitophilus species, especially their doses not causing high mortality.     

Bioconversion of 1-adamantanol to 1,3-adamantanediol using Streptomyces sp. SA8 oxidation system

To efficiently produce 1,3-adamantanediol (1,3-ad(OH)₂) from 1-adamantanol (1-adOH), our stocks of culture strains and soil microorganisms were surveyed for hydroxylation activity towards 1-adOH. Among them, the soil actinomycete SA8 showing the highest hydroxylation activity was identified as Streptomyces sp. based on 16S ribosomal DNA sequence analysis. The reaction products were purified by silica gel column chromatography, and from NMR and MS analyses, they were identified as 1,3-ad(OH)₂ and 1,4-ad(OH)₂. Streptomyces sp. SA8 produced 5.9 g l^? 1 1,3-ad(OH)₂from 6.2 g l^? 1 1-adOH in culture broth after 120 h at 25 °C. Using resting cells, 2.3 g l^? 1 1,3-ad(OH)₂ was produced after 96 h of incubation at a 69% conversion rate. In both cases, 1,4-ad(OH)₂ was formed as a byproduct at a rate of about 15%. Strain SA8 also hydroxylated 2-adamantanol and 2-methyl-2-adamantanol.     

Phytonutrient,Antioxidant and Mineral Composition of Some Wild Fruits in South West Nigeria

This study was carried out to determine the chemical composition, antinutrient and phytonutrient contents of some wild fruits, namely African star apple (Chrysophyllum albidum G. Don.), hog plum (Spondias mombin Linn.), bush mango (Irvingiagabonensis Baill) and monkey cola (Colamillenii K. Schum). Samples of the wild fruit commonly consumed were collected from some villages in Ido Local Government Area, Oyo State Nigeria. The wild fruits were analysed for phytonutrients, antioxidants and mineral composition. Ranges of total phenolics and total anthocyanin content of these wild fruits were 27.78 ± 6.01 in I. gabonensis, 57.42 ± 4.47 in S. mombin, 121.29 ± 4.97 in C. millenii and 398.23 ± 0.00 in C. albidum respectively. Significant differences (p < 0.05) were found in antinutrient, phytonutrient and mineral composition of C. albidum, I. gabonensis and C. millenii. The analysis of variance revealed that calcium and manganese contents of I. gabonensis were significantly (p < 0.05) higher than those of C. albidum and C. millenii. The antioxidants especially vitamin C content ranged from as low as 15.87 mg/100 g in C. albidum to as high as 204.86 mg/100 g in S. mombin. The values for total carotenoid ranged from 172.77 μg/100 g (in C. millenii) to 1380.17 μg/100 g (in C. albidum). The wild fruits are sources of phytonutrients, antioxidants such as vitamin C, total carotenoids and some minerals. Planting of the wild fruit trees or the incorporation in farming systems should thus be encouraged to increase production and availability to consumers and as industrial raw materials.     

Tenderization and fragmentation of myofibrillar proteins in bovine longissimus dorsi muscle using proteolytic extract from Sarcodon aspratus

The objective of this study was to examine the Sarcodon aspratus extract including protease how to affect tenderness of the bovine longissimus dorsi muscle. In addition, we investigated myofibrillar protein fragmentation, particularly in myosin, and its influence on meat tenderness. Beef loin chunks were marinated with 0.5 g/100 g, 1 g/100 g, and 2 g/100 g powdered S. aspratus extract, 0.2 g/100 mL papain, and distilled water (control), respectively. Although tenderness of meat is increased by adding S. aspratus extract, differences in meat quality traits, such as muscle pH and meat color, were small and not considered to have practical importance between the control and enzyme-treated samples. Furthermore, the S. aspratus extract influenced the myofibril fragmentation index (MFI) as well as protein solubility. The changes in MFI and protein solubility were due to the myofibrillar protein degradation. Through Western blotting, we found that the S. aspratus extract, as well as papain, caused fragmentation of the myosin heavy chain, but the mushroom extract induced more fragmentations of myofibrillar proteins, and caused more tender meat.     

Efficacy of filter cake and Triplex powders from Ethiopia applied to wheat against Sitophilus zeamais and Sitophilus oryzae

The efficacy of filter cake and Triplex powders applied to wheat was evaluated in the laboratory against the maize weevil, Sitophilus zeamais Motschulsky and rice weevil, Sitophilus oryzae (Linnaeus)—two most common insect pests associated with stored grain in Ethiopia. Efficacy of these powders was determined by exposing 20 adults of each species to 100 g of wheat treated with 0, 100, 500, 700 and 1000 mg/kg of filter cake and Triplex. Adult mortality was determined 7 and 14 d after exposure. In addition, adult progeny production, percentage of insect damaged kernels, and percentage of grain weight loss at each species-powder-concentration-time combinations were determined after 42 d. The 7 and 14 d mortality was 100% for adults of both species exposed to 1000 mg/kg of filter cake; only the 14 d mortality of Sitophilus species was 100% for adults exposed to 700 mg/kg. Mortality of S. oryzae adults was 100% when exposed for 14 d to1000 mg/kg of Triplex. Mortality of S. zeamais never reached 100% in any Triplex treatments. Adult progeny production of S. zeamais was completely suppressed at filter cake concentrations of 700 and 1000 mg/kg, whereas 1000 mg/kg was necessary for complete suppression of S. oryzae adult progeny production. Complete suppression of adult progeny production was not observed in any Triplex treatments. Complete reduction in percentage of insect damaged kernels and percentage of grain weight loss were obtained when S. zeamais and S. oryzae adults were exposed to 1000 mg/kg of filter cake; similar reductions with S. zeamais occurred only at 1000 mg/kg of Triplex. In the case of S. oryzae, complete reduction of insect damaged kernels and grain weight loss were not achieved at any concentration of Triplex. These powders can be used as alternatives to chemical insecticides for management of Sitophilus species.     

Rapid and reliable identification of Staphylococcus aureus harbouring the enterotoxin gene cluster (egc) and quantitative detection in raw milk by real time PCR

A TaqMan and a SYBR Green real time PCR (rt-PCR) were developed for the reliable identification and quantitative detection of Staphylococcus (S.) aureus strains harbouring the enterotoxin gene cluster (egc) regardless of its variants. Both approaches revealed 100% specificity against a panel of 70 reference strains, including 29 clinical and foodborne S. aureus strains harbouring all the egc variants to date known, 4 egc⁻S. aureus strains and 37 strains of phylogenetically closely and distantly related species. Standard curves made by 10 fold dilutions of either genomic DNA or cells from an egc⁺S. aureus log-phase broth culture showed a good linearity of response (R² ≥ 0.993) for six orders of magnitude, with about 100% relative accuracy and a low inter-assay variability (CV ≤ 3.02). The overall limit of quantification (LOQ) for both rt-PCR assays (about 100% PCR efficiency; running time 30 min) was 10 cfu or 10 genome equivalents per reaction mixture although 1 cfu or 1 genome equivalent was detected with a 33.33% probability. These performances were confirmed in raw milk artificially contaminated with log-phase broth cultures of either a single egc⁺S. aureus strain or a mixture of S. aureus strains harbouring all the egc variants to date known. Similar results were also obtained with a raw milk based standard curve of the S. aureus egc⁺ mixture in the presence of 10⁶ cfu/mL of egc⁻S. aureus strains harbouring some of the commonest enterotoxin genes associated to the staphylococcal food poisoning. Nonetheless, the TaqMan based approach resulted in a lower sensitivity (LOQ = 100 cfu equivalents per reaction mixture) than the SYBR Green based assay (LOQ = 10 cfu equivalents per reaction mixture). When applied to real milk samples, both PCR assays provided a good response with 100% diagnostic specificity and 96-107% relative accuracy, as compared to conventional culture-based PCR approaches. Due to the high specificity, the wide dynamic range of detection and the high sensitivity demonstrated even in a complex and potentially highly contaminated raw milk matrix, the SYBR Green rt-PCR assay is a useful diagnostic tool for quick, high throughput and reliable routine screening of egc⁺S. aureus isolates. Moreover, the SYBR Green based quantitative detection of these pathogens in raw milk could remarkably contribute to clarify their actual role in staphylococcal food poisoning and other clinical syndromes associated with the consumption of milk and milk-based products.     

Investigation of the pathophysiology of bacterial mastitis using precision-cut bovine udder slices

Mastitis in cattle is a major health problem as well as incurring high costs for the dairy industry. To assess the suitability of precision-cut bovine udder slices (PCBUS) for bovine mastitis studies, we infected PCBUS with 2 different Staphylococcus aureus strains. Accordingly, we investigated both the tissue response to infection based on immune mediators at the mRNA and protein levels and the invasion of bacteria within the tissue. The studied proteins represent immune mediators of early inflammation [IL-1β, tumor necrosis factor-α (TNF-α), prostaglandin E₂ (PGE₂)] and showed a time-dependent increase in concentration. Infection of PCBUS with S. aureus resulted in increased expression of proinflammatory cytokines and chemokines such as TNF-α, C-C motif chemokine ligand 20 (CCL20), IL-1β, IL-6, and IL-10, but not C-X-C motif chemokine ligand 8 (CXCL8), lingual antimicrobial peptide (LAP), or S100 calcium binding protein A9 (S100A9) at the mRNA level. To compare the data acquired with this model, we carried out investigations on primary bovine mammary epithelial cells. Our results showed that the immune responses of both models—PCBUS and primary bovine mammary epithelial cells—were similar. In addition, investigations using PCBUS enabled us to demonstrate adherence of bacteria in the physiological cell network. These findings support the use of PCBUS in studies designed to further understand the complex pathophysiological processes of infection and inflammation in bovine mastitis and to investigate alternative therapies for mastitis.     

The spatial expression pattern of antimicrobial peptides across the healthy bovine udder

Antimicrobial peptides are key molecules in local host defense. With the aim to better understand the possible involvement of these peptides in the prevention of bovine mastitis, we determined, for the first time to our knowledge, the spatial pattern of constitutive expression of 5 bovine β-defensins and bovine psoriasin (S100A7) across 5 localizations of the bovine mammary gland applying a quantitative real-time PCR approach. We observed 3 different expression patterns in the healthy udder: 1) constitutive expression of the lingual and tracheal antimicrobial peptides (LAP and TAP), as well that of bovine neutrophil β-defensins 4 and 10 (BNBD4 and BNBD10), is essentially restricted to the mammary lymph node; 2) bovine β-defensin 1 (DEFB1) is mainly expressed in the cisternal epithelium and the Rosette of Fürstenberg; 3) strong constitutive mRNA expression of the calcium-binding protein S100A7, which is also known as psoriasin and which has been reported to be highly active against Escherichia coli, was detected in the streak canal. These results indicate a crucial role of S100A7 in the early-stage prevention of coliform mastitis, and the analyzed β-defensins might be regarded as inducible weapons against already invading pathogens.     

Physicochemical and structural characterization of Lima Bean (Phaseolus lunatus) globulins

A globulin fraction from Phaseolus lunatus have been isolated and characterized. The fractionation scheme used allowed to obtain a Globulin partially purificated. Globulin purification by ultracentrifugation allowed to isolate three peaks with different sedimentation coefficient, while gel filtration chromatography yielded two species, I and II, which were later identified as 11S and 7S globulins, respectively. Species II, 7S globulin from P. lunatus, seems to have a molecular mass of 72 kDa and to be constituted by polypeptides of 34-36, 25-27 and 16-18 kDa without intermolecular disulfide bonds. These polypeptides would present certain heterogeneity, as suggested by the results of isoelectric focusing and electrophoresis analyses. This protein exhibited some particular features, including a lower molecular mass and higher thermal stability than typical vicilins. Species I, with a molecular mass of 336 kDa, would be constituted by subunits of 53-55 and 40-41 kDa formed by smaller polypeptides (around 30 and 20 kDa) linked by disulfide bonds. These features, together with the existence of basic and acid polypeptides in the molecule, would confirm that species I is a globulin of the legumins, 11S globulins or α-conglutin family.     

杏鲍菇分离蛋白和清蛋白的理化性质及功能分析

以杏鲍菇为原料,分别利用碱溶酸沉法提取杏鲍菇分离蛋白（Pleurotus eryngii protein isolate,PEPI）、Osborne法分离主要蛋白组分,并研究其理化性质和功能分析。结果表明,杏鲍菇的蛋白质量分数为17.57%（以干质量计）,以杏鲍菇清蛋白（Pleurotus eryngii albumin,PEA）为主,占总分离蛋白组分的81.12%。PEPI和PEA中均含18?种氨基酸,且必需氨基酸含量分别占总氨基酸含量的40.80%和40.51%。与PEPI相比,PEA的表面疏水性（265.25）显著高于PEPI（164.27）（P＜0.05）,而总巯基、二硫键含量较低,分别为61.53?μmol/g和10.39?μmol/g;热变性温度（100.98?℃）低于PEPI（108.27?℃）,且PEA持水性（1.64?mL/g）、持油性（5.59?mL/g）显著低于PEPI（3.58、8.36?mL/g）（P＜0.05）。PEPI和PEA的溶解性、起泡性、泡沫稳定性、乳化性及乳化稳定性随pH值的变化趋势均相似,在等电点时均为最低。傅里叶变换红外光谱显示PEPI和PEA的二级结构主要是β-折叠和β-转角,扫描电镜观察PEPI呈蜂巢结构。相比PEA,PEPI具有更好的理化性质和功能特性。     

Phenotypic and genotypic aspects of Lactobacillus sanfranciscensis strains isolated from sourdoughs in Italy

Physiological properties and genetic characteristics of 44 strains of Lactobacillus sanfranciscensis, isolated in Italy from sourdough samples for salted and sweet baked products, were investigated. Data were compared with those obtained for reference strains belonging to the same and to related species. Phenotypic analysis showed the presence of two main groups, the first including strains able to ferment glucose and maltose, the second including strains fermenting only maltose. Other few isolates exhibited individual different fermentation patterns. Genetic diversity was evaluated by using Amplified Ribosomal 16S rDNA Restriction Analysis and Amplified 16S-23S rDNA Spacer Region Analysis. Electrophoretical patterns of fragments generated by Hinf I indicated a polymorphism within 16S rDNA gene that allowed the subdivision ofLb. sanfranciscensis strains in two groups. The reliability to use Dde I and Hinf I 16S rDNA restriction analysis for differentiation of Lb. sanfranciscensis strains from the other phylogenetically related Lactic Acid Bacteria was demonstrated. The amplification of the intergenic spacer region produced a unique electrophoretical profile of three bands (approximately 300, 390 and 580 bp) for all Lb. sanfranciscensis strains; this analysis proved to be helpful for the discrimination of sourdough lactobacilli at species level, but was unable to distinguish among the isolates belonging to the same species. Results of grouping, obtained by genetic analyses, did not agree with those attained with phenotypic analysis, and no relation with the geographic area of isolation or type of product was found.     

Histamine content and histamine-forming bacteria in dried milkfish (Chanos chanos) products

Thirty-two dried milkfish products sold in five retail markets in southern Taiwan were purchased and tested to determine the occurrence of histamine and histamine-forming bacteria. Except for histamine and cadaverine, the average content of various biogenic amines in tested samples was less than 8.5 mg/100 g. Most of the tested dried milkfish products (78.1%) had histamine levels greater than the FDA guideline of 5 mg/100 g for scombroid fish and/or product, while fourteen of them (43.7%) contained > 50 mg/100 g hazard action level. Thirty histamine-producing bacterial strains, capable of producing 5.4 ppm to 562 ppm of histamine in trypticase soy broth supplemented with 1.0% L-histidine (TSBH), were identified as Enterobacter aerogenes (seven strains), Citrobacter sp. (one strain), Staphylococcusxylosus (ten strains), S. sciuri (one strain), Bacillus thuringiensis (two strains), Citrobacter freundii (five strains), Klebsiella pneumoniae (one strain) and E. cloacae (three strains), by 16S rDNA sequencing with PCR amplification.     

High prevalence of Salmonella enterica subsp. diarizonae in tonsils of sheep at slaughter

The presence of Salmonella was studied in tonsils and feces of sheep and goats at slaughter using PCR and culturing. The isolates were further characterized using PFGE to get more information about the genetic diversity of Salmonella strains circulating among sheep and goats. Antimicrobial resistance was studied because resistance to multiple antimicrobial agents among Salmonella is increasing. The prevalence of Salmonella was 43% and 2% in the tonsils of sheep and goat, respectively. Salmonella was not detected in the feces of adult animals and only sporadically in the feces of juveniles (2%). S. enterica subsp. diarizonae 61:k:1,5,(7) was isolated from 20% of the sheep tonsils and 1% of the goat tonsils. In total, 9 genotypes were obtained with PFGE using SpeI, XbaI, NotI and XhoI restriction enzymes; however, one genotype was predominant. All strains were sensitive to most (13/16) of the antimicrobials. Resistance to sulfamethoxazole was high (95%). Three (15%) strains, which were isolated from lambs, were also resistant to colistin. No correlation between the antimicrobial resistance pattern and the genotype was noticed. These results demonstrate that slaughtered sheep are an important reservoir for S. enterica subsp. diarizonae 61:k:1,5,(7) carrying this pathogen frequently in the tonsils. Future studies are needed to elucidate the significance of the tonsils in the contamination of sheep carcasses and meat with Salmonella.     

Gone with the weed: Population growth of Sitophilus oryzae and Rhyzopertha dominica in wheat and barley containing seeds of Silybum marianum

In the present study we examined the effect of seeds of the weed Silybum marianum (L.) Gaertn. (Asterales: Asteraceae) in different combinations with wheat or barley in the population growth of two major stored product insects, Rhyzopertha dominica (F.) (Coleoptera: Bostryhidae) and Sitophilus oryzae (L.) (Coleoptera: Curculionidae). The evaluation took place on 7 different quantitative combinations of the weed, i.e. 0, 1, 5, 10, 20, 50 and 100% of the total amount of the grain mass. All combinations were kept under constant conditions of 25 ^οC and 65% relative humidity (r.h.) for 65 days. After this interval, adult progeny production was counted, and classified as dead or alive. In general, progeny production was higher on wheat than on barley for R. dominica, but the reverse was recorded for S. oryzae. No progeny production was recorded for either species when S. marianum containment was 100%. Moreover, for both species, the decrease of the percentage of S. marianum caused an increase in progeny production, but this decrease was not linear. The study concludes that R. dominica and S. oryzae cannot develop on S. marianum seeds, but they can develop in mixtures of these seeds with grains.     

Potential of three enhanced diatomaceous earths against Sitophilus zeamais Motschulsky and Prostephanus truncantus (Horn) on stored maize grain

Global health and environmental concerns against synthetic pesticides in stored-grain coupled with high cost, unavailability and inaccessibility of modern non-chemical grain treatment technologies in Africa are driving the need for low-cost and low-dose combination of readily available alternatives. Laboratory studies were conducted to determine the efficacy of non-registered low-dose food grade diatomaceous earths enhanced with natural grain protectants: IBS at 50, 75 and 100 ppm; IPE1 at 200, 300 and 400 ppm; IPE2 at 100, 150 and 200 ppm against Prostephanus truncatus and Sitophilus zeamais adult insects on maize. Adult insect mortality was assessed at 7, 14 and 21 days after treatment (DAT) while progeny emergence was assessed at 49 and 70 DAT for S. zeamais and P. truncatus, respectively. IBS caused significant (p < 0.05) P. truncatus and S. zeamais adult mortality at 14 and 21 DAT, respectively. Except for IPE1 at 400ppm, lower doses of IPE1 and IPE2 showed significantly low adult mortality (p < 0.05) (54-74%) on Sitophilus zeamais compared to IBS and the positive control. All rates of IPE1 and IPE2 were also not effective on P. truncatus adults, with significantly low mortality (p < 0.05) (18–38% compared to positive control). IBS at 75 and 100 ppm were effective on progeny suppression on both S. zeamais and P. truncatus but IPE1 at 400 ppm was effective on S. zeamais only. These results suggest the potential of IBS and IPE1 (at 400 ppm) as effective sustainable alternatives to synthetic grain protectants for small-scale farmers in sub-Saharan Africa who may not afford the modern non-chemical grain protection technologies. Field efficacy of these materials under typical farmer conditions is required.     

Effects of variousMicrococcus strains on the ripening and organoleptic characteristics of Arzúa cow's-milk cheese

Ten batches of Arzúa, a soft cow's-milk cheese produced in northwest Spain, were prepared from pasteurized milk. Two (control) batches (CB) were made with a commercial starter containingLactococcus lactis subspecieslactis andcremoris. Another eight batches (MB) were made with the commercial starter plus one of eightMicrococcus spp. strains previously isolated from raw-milk Arzúa cheeses. In all MB, β-casein degradation over the 30-day ripening period was more pronounced (mean 31%) than in the CB (mean 12%). α_S1-Casein degradation was highly variable in the MB, though mean degradation over the ripening period (75%) was similar to that observed in the CB (73%). Similarly, volatile fatty acid (VFA) content was highly variable in the MB, with the mean content at 30 days (3.8 mEq per 100 g) being higher than in the CB (1.6 mEq per 100 g). Rheological characterization of the cheeses (by uniaxial compression) revealed significant differences between batches, with some samples fracturing under the compression pressure applied and others not. Sensory evaluation also revealed significant differences. “Non-milk” aromas were more frequently detected in batches made with lipolytic micrococcal strains. Betweenbatch differences in tastes and texture were also detected. Multiple correlation analysis of the data obtained at day 15 of ripening revealed statistically significant positive correlations (r>0.70) between α_S1-casein content and dry matter content, between α_S1-casein content and sensorially evaluated firmness, and between VFA content and sensorially evaluated rancidity. Statistically significant negative correlations between sensorially evaluated firmness and the ratio of α_S1-I content to α_S1-casein content were detected. The results of this study suggest a need for further studies aimed at selecting those strains which could be most suitable for the production of Arzüa cheeses; due to their effects on texture, α_S1-caseinolytic strains seem to be more appropriate than β-caseinolytic ones.     

Comparative chemical composition,free radical-scavenging and cytotoxic properties of essential oils of six Stachys species from different regions of the Mediterranean Area

The chemical composition of essential oils of six Stachys species, S. cretica L. ssp. vacillans Rech. fil., S. germanica L., S. hydrophila Boiss., S. nivea Labill., S. palustris L. and S. spinosa L., obtained by hydrodistillation, was studied by GC and GC–MS. All the oils have in common a great percentage of fatty acids and esters (24.2–58.5%) and a high amount of sesquiterpenes (16–35.9%), with the exception of the oil from S. palustris, which consisted mainly of carbonylic compounds (25.4%). The antioxidant activity by DPPH test and the antiproliferative activity on a series of human cancer cell lines (C32, amelanotic melanoma and ACHN, renal cell adenocarcinoma) were investigated for all the oils. S. palustris,S. cretica and S. hydrophila showed the highest antiradical effect, with IC₅₀ values of 0.482, 0.652 and 0.664 mg/ml, respectively. The most antiproliferative essential oil against C32 cell line was the oil of S. germanica with a 77% of inhibition at a concentration of 100 μg/ml. S. germanica, S. palustris and S. spinosa showed the most antiproliferative activity on ACHN cell line, at a concentration of 100 μg/ml,with 81%, 77% and 73% inhibition, respectively.