Nutrient deficiency in the production of artemisinin, dihydroartemisinic acid, and artemisinic acid in Artemisia annua L

Artemisia annua became a valuable agricultural crop after the World Health Organization recommended artemisinin as a component of ACT (artemisinin-combination based therapies) for malaria in 2001. A cloned, greenhouse-grown, A. annua (Artemis) subjected to an acidic soil and macronutrient deficit was evaluated for artemisinin production. Lack of lime (L) and macronutrients (N, P, and K) reduced leaf biomass accumulation. When L was provided (pH 5.1), the highest average leaf biomass was achieved with the "complete" (+N, +P, +K, and +L) treatment (70.3 g/plant), and the least biomass was achieved with the untreated (-N, -P, -K, and -L) treatment (6.18 g/plant). The nutrient least required for biomass accumulation per plant (g) was K (49.0 g), followed by P (36.5 g) and N (14.3 g). The artemisinin concentration (g/100 g) was significantly higher (75.5%) in -K plants when compared to plants under the complete treatment (1.62 vs 0.93%). Although the artemisinin total yield (g/plant) was 21% higher in -K plants, it was not significantly different from plants under the complete treatment (0.80 vs 0.66 g/plant). There were no marked treatment effects for concentration (g/100 g) or yield (g/plant) of both dihydroartemisinic acid and artemisinic acid, although higher levels were achieved in plants under the complete or -K treatments. There was a positive and significant correlation between artemisinin and both artemisinic acid and dihydroartemisin acid, in g/100 g and g/plant. This is the first report where potassium deficiency significantly increases the concentration (g/100 g) of artemisinin. Thus, under a mild potassium deficiency, A. annua farmers could achieve similar gains in artemisinin/ha, while saving on potassium fertilization, increasing the profitability of artemisinin production.     

Structural ripening-related changes of the arabinan-rich pectic polysaccharides from olive pulp cell walls

In this study, the structural features and ripening-related changes that occur in the arabinan-rich pectic polysaccharides highly enmeshed in the cellulosic matrix of the olive pulp fruit were evaluated. These pectic polysaccharides, obtained from two consecutive harvests at green, cherry, and black ripening stages, account for 11-19% of the total pectic polysaccharides found in the olive pulp cell walls and were previously shown to occur as calcium chelating dimers. On the basis of the 13C NMR, (1H, 13C) gHSQC, 2D COSYPR, and (1H,13C) gHMBC carbon and proton resonances of the variously linked arabinosyl residues, we propose a tentative structure. This structure is particularly characterized by T-beta-Araf (1-->5)-linked to (1-->3,5)-Araf residues and by the occurrence of branched and linear blocks in the arabinan backbone. Methylation analysis showed that these pectic polysaccharides of black olives have more arabinan side chains, which were shorter (less (1-->5)-Araf), highly branched (more (1-->3,5)-Araf), and with shorter side chains (fewer (1-->3)-Araf) than those of green and cherry olives. Quantitative 13C NMR data indicated that these modifications involved the disappearance of the characteristic terminally linked beta-Araf residue of the arabinans. This odd feature can be used as a diagnostic tool in the evaluation of the stage of ripening of this fruit, as well as a marker for the presence of olive pulp in matrices containing pectic polysaccharides samples.     

Inhibition of trypsin by condensed tannins and wine

Phenolic compounds are abundant vegetable secondary metabolites in the human diet. The ability of procyanidin oligomers and wine polyphenols to inhibit trypsin activity was studied using a versatile and reliable in vitro method. The hydrolysis of the chromogenic substrate N-benzoyl-d,l-arginine-p-nitroanilide (BApNA) by trypsin was followed by spectrophotometry in the presence and absence of condensed tannins and wine. A clear relationship between the degree of polymerization of procyanidins and enzymatic inhibition was observed. Trypsin activity inhibition was also detected in several types of wine. In general, the inhibition increased with the concentration of phenolic compounds in wines. These results may be relevant when considering these compounds as antinutritional factors, thereby contributing to a reduced absorption of nutrients.     

Effects of growing conditions on purple corncob (Zea mays L.) anthocyanins

Purple corn ( Zea mays L.) has been used for centuries as a natural food colorant in South America and, more recently, in Asia and Europe. However, limited information is available on the factors affecting their anthocyanin concentration and profiles. In this study, 18 purple corn samples grown under different conditions in Peru were evaluated for quantitative and qualitative anthocyanin composition as well as total phenolics. High variability was observed on monomeric anthocyanin and phenolic contents with yields ranging from 290 to 1333 mg/100 g dry weight (DW) and from 950 to 3516 mg/100 g DW, respectively, while 30.5-47.1% of the total phenolics were anthocyanins. The major anthocyanins present were cyanidin-3-glucoside, pelargonidin-3-glucoside, peonidin-3-glucoside, cyanidin-3-maloylglucoside, pelargonidin-3-maloylglucoside, and peonidin-3-maloylglucoside, and 35.6-54.0% of the anthocyanins were acylated. Potassium sources/concentrations on the soil and seedling density did not significantly affect anthocyanin composition. The growing location affected anthocyanin levels and the percentage of anthocyanins to total phenolics ( p < 0.01) and should be taken into account when choosing a material for color production.     

Less is more: Little seed processing required for direct seeding in seasonal tropics

Seed processing and dormancy break treatments are recommended for rendering seeds restoration-ready. Conversely, fruit structures and seed coats may protect seeds from environmental harm in the field. We evaluated the effects of seed processing (by either keeping fruit structures or extracting seeds) and/or scarification (of physically dormant seeds) on the seedling emergence and establishment of 10 legume tree species from tropical forests and savannas of Central Brazil. We sowed seeds in a greenhouse for reference and in a field experiment conducted in tilled ready-to-seed sites. We monitored seedling emergence and survival for a year. We calculated the costs of harvesting, processing, and pretreating seeds, and considered the final cost of a 1-year-old seedling. Seed extraction resulted in lower emergence for most species in the greenhouse and in the field. It also accelerated emergence of three and four species in the greenhouse and the field, respectively. Scarification resulted in lower seedling emergence in the field for half of the species, while it increased emergence of three species in the greenhouse. Most species presented accelerated emergence both in the greenhouse and the field. The seedling cost was 1.6 to 74.6 times higher when seeds were processed, and 1.3 to 6.0 times when seeds were scarified, except for one species. Keeping fruit structures and seed coats reduced the costs of seeds and increased the success of direct seeding.

     

Porcine circovirus type 2 decreases the infection and replication of attenuated classical swine fever virus in porcine alveolar macrophages

Recently, it has been noted that porcine circovirus type 2 (PCV2) infection adversely affects the protective efficacy of Lapinized Philippines Coronel (LPC) vaccine, an attenuated strain of classical swine fever virus (CSFV), in pigs. In order to investigate the possible mechanisms of the PCV2-derived interference, an in vitro model was established to study the interaction of LPC virus (LPCV) and PCV2 in porcine alveolar macrophages (AMs). The results showed that PCV2 reduced the LPCV infection in AMs and the levels of PCV2-derived interference were dose-dependent. The PCV2-derived interference also reduced the replication level of LPCV in AMs. The full-length PCV2 DNA and its fragment DNA C9 CpG-ODN were involved in the reduction of LPCV infection in AMs, whereas UV-inactivated PCV2 was not. In addition, a moderate negative correlation between the LPCV antigen-containing rate and IFN-γ production was observed, and had a dose-dependent trend with the level of PCV2-inoculation. The results of the present study may partially explain how PCV2 infection interferes with the efficacy of LPC vaccine.     

Detection of retinoid receptors in non-neoplastic canine lymph nodes and in lymphoma

This study evaluated the difference in retinoid receptor expression between non-neoplastic lymph nodes and nodal lymphoma in dogs. Retinoid receptor expression was evaluated by immunohistochemistry in 32 canine lymph nodes. The lymph nodes had been previously diagnosed as non-neoplastic (6 normal and 7 hyperplastic lymph nodes) and B- and T-cell lymphoma (19 cases). Immunohistochemistry for retinoic acid receptors and retinoid-X receptors (and their subtypes α, β, and γ) was performed in all cases. In addition, immunohistochemistry for CD3 and CD79a was performed in all lymphoma cases. Non-neoplastic lymphocytes were negative for all retinoid receptors. Retinoic acid receptor-γ was detected in 100% of B-cell lymphoma and 78% of T-cell lymphoma, while retinoid X receptor-γ was positive in 78% of T-cell lymphoma cases. When normal lymph node architecture was still present, a contrast between retinoid-negative benign cells and retinoid-positive malignant cells was clear. Retinoid receptors were expressed in neoplastic, but not in benign lymphocytes, suggesting their value for both diagnosis and treatment of canine lymphoma.     

Effects of Artemisia annua and Foeniculum vulgare on chickens highly infected with Eimeria tenella (Phylum Apicomplexa)

Background

Intensive poultry production systems depend on chemoprophylaxis with anticoccidial drugs to combat infection. A floor-pen study was conducted to evaluate the anticoccidial effect of Artemisia annua and Foeniculum vulgare on Eimeria tenella infection. Five experimental groups were established: negative control (untreated, unchallenged); positive control (untreated, challenged); a group medicated with 125 ppm lasalocid and challenged; a group medicated with A. annua leaf powder at 1.5% in feed and challenged; and a group treated with the mixed oils of A. annua and Foeniculum vulgare in equal parts, 7.5% in water and challenged. The effects of A. annua and oil extract of A. annua + F. vulgare on E. tenella infection were assessed by clinical signs, mortality, fecal oocyst output, faeces, lesion score, weight gain, and feed conversion.

Results

Clinical signs were noticed only in three chickens from the lasalocid group, six from the A. annua group, and nine from the A. annua + F. vulgare group, but were present in 19 infected chickens from the positive control group. Bloody diarrhea was registered in only two chickens from A. annua group, but in 17 chickens from the positive control group. Mortality also occurred in the positive control group (7/20). Chickens treated with A. annua had a significant reduction in faecal oocysts (95.6%; P = 0.027) and in lesion score (56.3%; P = 0.005) when compared to the positive control. At the end of experiment, chickens treated with A. annua leaf powder had the highest body weight gain (68.2 g/day), after the negative control group, and the best feed conversion (1.85) among all experimental groups.

Conclusions

Our results suggest that A. annua leaf powder (Aa-p), at 1.5% of the daily diet post-infection, can be a valuable alternative for synthetic coccidiostats, such as lasalocid.     

Effects of road transportation or droving on the weight and metabolism of young bulls

This study evaluated the effects of the mode of transportation on weight and metabolism of young bulls, which are generally transported by two systems: droving and truck. This experiment consisted of two phases: transportation and refeeding and involved transportation by droving and truck. Fifty young Nellore bulls were separated in equal numbers in both groups, droving group and truck group. In the transportation phase, animals were moved, in a 640-km journey. The refeeding phase consisted of 84 days and commenced after the animals arrived at the fattening farm. The effect of the transportation system on animals’ weight, metabolic hormones, and serum biochemistry was assessed after 28, 56, and 84 days. During the transportation phase, animals in the droving group lost 10.11 kg and animals in the truck group gained 13.1 kg. The truck group showed 20.5 kg of weight gain at the end of the refeeding phase. The highest triiodothyronine (T₃) and thyroxine (T₄) concentrations were observed on days 0 and 60 in the transportation phase in both droving and truck groups. The serum cortisol concentration was higher in the droving group than in the truck group on day 56 of the refeeding phase. The IGF-I concentrations were higher during the refeeding phase in the droving group than in the truck group. Our results showed that transportation by droving leads to increased weight loss and reduced further weight gain compared to transportation by truck and that serum concentrations of T₄, cortisol, and insulin-like growth factor (IGF-I) hormones are altered by the transportation system.     

Evaluation of hepatitis E virus infection between different production systems of pigs in Brazil

The objectives of this study were to (1) investigate the occurrence of hepatitis E virus (HEV) in pigs from large-scale and family-scale farms, (2) genetically characterize the strains isolated, and (3) study the pathogenesis of swine HEV infection via immunohistochemistry. A total of 50 pigs from 10 farms in Mato Grosso State, Brazil were divided according to type of production system into either large-scale farms (n?=?5) or family-scale farms (n?=?5). Samples of liver, gallbladder, small and large intestines, bile, and feces from the pigs were analyzed by nested PCR with primers targeting the ORF2 region of HEV and by immunohistochemistry. Of the eight HEV-positive samples from pigs of family-scale farms, phylogenetic analysis revealed that seven of the swine HEV isolates clustered with subtype 3b of genotype 3 and one isolate was categorized with subtype 3 f. The HEV antigen was detected mainly in the small intestine samples from family-scale farms, suggesting an early stage HEV infection. HEV was not detected in the samples of pigs from large-scale farms, reinforcing the need for additional studies to evaluate the risk of transmission of HEV to humans from pigs from family-scale farms in Mato Grosso State.