GC-MS metabolite profiling of <Emphasis Type="Italic">Phytophthora infestans</Emphasis> resistant to metalaxyl

Phytophthora infestans is the most important potato pathogen worldwide. Various alternatives have been used to control the pathogen, including continuous applications of phenylamide fungicides which has caused a rapid development of resistance in populations of P. infestans. Despite the importance of the disease, metabolite profiling of fungicide-resistant P. infestans has not been reported. In vitro resistance of Phytophthora infestans isolates to metalaxyl was characterized and metabolic changes in resistant isolates were evaluated at low (0.5 mg/L) and high (100 mg/L) concentrations of the fungicide. About 70% of the isolates tested showed resistance to metalaxyl and a total of 49 metabolites were differently expressed in resistant isolates growing in the presence of the fungicide. Principal components analysis revealed a distinct metabolite profile of resistant isolates exposed to both low and high levels of metalaxyl. The main metabolites responsible for the clustering in both fungicide concentrations included fatty acids such as hexadecanoic and octadecanoic acids, sugars such as glucose and fructose, aminoacids such as proline and valine, and organic acids such as butanedioic and propanoic acids. Potential resistance-related metabolic pathways are mostly involved in the regulation of the pathogen’s membrane fluidity and included the fatty acid biosynthesis as well as the glycerophospholipid metabolism pathways. This is the first metabolomic-based characterization of fungicide resistance in plant pathogens.     

Production of Bihon-type Noodles from Maize Starch Differing in Amylose Content

Maize starches extracted from selected maize cultivars with 0.2–60.8% amylose contents were used to produce bihon-type noodles. Starch dough using a pregelatinized starch binder was prepared and extruded through a laboratory-scale extruder simulating the traditional process of making bihon in the Philippines. The normal maize starches with amylose content of ≈28% were successfully used for bihon-type noodle production, but waxy maize starches with 0.2–3.8% amylose content and high-amylose maize starches with 40.0–60.8% amylose content failed to produce bihon-type noodles. Viscoamylograph profile parameters and swelling volume are significantly correlated to amylose content of maize starch samples evaluated. These physicochemical properties may be used to indicate that the starch samples at normal amylose levels may be used for bihon-type noodles. Starch noodles produced in the laboratory were not significantly different in terms of either cooking quality or textural properties from two commercially produced maize noodle samples, except for adhesiveness. The laboratory process and fabricated extruder can be used to produce bihon-type noodles.     

Phytochemische Untersuchungen an Pflanzen der kubanischen Flora

Zusammenfassung In Zusammenarbeit zwischen der Akademie der Wissenschaften Kubas und der Deutschen Akademie der Wissenschaften zu Berlin wurden 1165 Pflanzenarten aus 149 Familien der kubanischen Flora nach Standardverfahren orientierend auf Alkaloide und Saponine untersucht. Die Ergebnisse sind in Tabelle 2 aufgeführt.

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Phytochemical investigations in plants of the Cuban flora

Summary In cooperation of the Academy of Sciences of Cuba with the German Academy of Sciences at Berlin 1165 species from 149 plant families of the Cuban flora have been preliminarily investigated by standard methods with regard to the occurrence of alkaloids and saponins. The results are compiled in table 2.

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1165 149 . . . 2.

     

Pharmacokinetics of enrofloxacin HCl‐2H2O (ENRO‐C), PK/PD,and Monte Carlo modeling vs. Leptospira spp. in cows

The pharmacokinetics, PK/PD ratios, and Monte Carlo modeling of enrofloxacin HCl‐2H₂O (Enro‐C) and its reference preparation (Enro‐R) were determined in cows. Fifty‐four Jersey cows were randomly assigned to six groups receiving a single IM dose of 10, 15, or 20 mg/kg of Enro‐C (Enro‐C₁₀, Enro‐C₁₅, Enro‐C₂₀) or Enro‐R. Serial serum samples were collected and enrofloxacin concentrations quantified. A composite set of minimum inhibitory concentrations (MIC) of Leptospira spp. was utilized to calculate PK/PD ratios: maximum serum concentration/MIC (C_max/MIC₉₀) and area under the serum vs. time concentration of enrofloxacin/MIC (AUC_0‐24/MIC₉₀). Monte Carlo simulations targeted C_max/MIC = 10 and AUC_0‐24/MIC = 125. Mean C_max obtained were 6.17 and 2.46 μg/ml; 8.75 and 3.54 μg/ml; and 13.89 and 4.25 μg/ml, respectively for Enro‐C and Enro‐R. C_max/MIC₉₀ ratios were 6.17 and 2.46, 8.75 and 3.54, and 13.89 and 4.25 for Enro‐C and Enro‐R, respectively. Monte Carlo simulations based on C_max/MIC₉₀ = 10 indicate that only Enro‐C₁₅ and Enro‐C₂₀ may be useful to treat leptospirosis in cows, predicting a success rate ≥95% when MIC₅₀ = 0.5 μg/ml, and ≥80% when MIC₉₀ = 1.0 μg/ml. Although Enro‐C₁₅ and Enro‐C₂₀ may be useful to treat leptospirosis in cattle, clinical trials are necessary to confirm this proposal.     

Effect of prepubertal nutrition on cellular apoptosis and proliferation in at term placenta of Anglo‐Nubian goats

The aim of this study was to evaluate the effect of feed restriction with or without monensin supplementation, followed by a re‐feeding period on cellular apoptosis and proliferation in at term placenta of Anglo‐Nubian goats. To evaluate the induction of apoptosis through the intrinsic pathway, proteins Bax and Bcl‐2 were determinated. The apoptosis was related with the cell proliferation indices through Ki67 determination. The treatments were applied for 250 days and were (a) ad libitum feeding (control; n = 5); (b) restricted feeding at 70% of control (restricted; n = 7); and (c) restricted with monensin supplementation (monensin; n = 7). After treatments, all the animals were fed to support their requirements. After parturition, 27 placentas were gathered. The placental cellular structure was studied by high‐resolution light microscopy and transmission electron microscopy; the cellular proliferation was determined by Ki67 index, and Bax and Bcl‐2 proteins were localized by immunohistochemical analysis. Differences in cell proliferation through the Ki67 index were found in monensin group placentas. Monensin supplementation stimulated the placental cell proliferation reversing the effect of feed restriction during the peripuberal period. A significant increase of Bcl‐2 in placentas of restricted group was found, and it would provide a protective effect on the placental structure. A lack of the Bcl‐2 protective effect was observed in control and monensin group placentas, probably meaning that the observed apoptosis would be induced through the intrinsic signalling pathway. A balance between apoptosis and cell proliferation is necessary to maintain tissue homoeostasis during caprine placental development.     

Significance of a common extended mucosal interface observed in transverse small intestine sonograms

An echogenic stripe can be seen through the mucosal layer on either side of the ultrasonographic image of a bowel loop in cross-section. The mucosa is a continuous layer surrounding the intestinal lumen; thus it should appear as a continuous hypoechoic band with no bright stripe in it. The purpose of this study was to investigate the origin of this stripe and to determine whether it is an artifact. Intestinal cross-sectional images were obtained using different transducer types and frequencies. In vivo and in vitro, different angles of insonification were used and, in vitro, bowel loops were also fluid filled during an ultrasonographic examination, to assess the potential influence of intestinal shape and position on this stripe. Some of these loops were evaluated histologically to determine if the echogenic stripe had a histologic basis. The echogenic stripe was present only when the loop was flattened. It remained on each side of the maximal cross-sectional width and disappeared when this width was parallel to the ultrasound beam, or when the bowel loop had a completely round shape and was dilated. There was no influence of transducer type, shape, or frequency on the appearance of the line. Histologically, uneven and larger distance between the mucosal villi could be seen on either side of the bowel loop corresponding to the location of the echogenic stripe seen on the ultrasonographic images. In conclusion, the occasional echogenic stripe represents an interface within the mucosa due to altered position of villi on either side of the maximal intestinal cross-sectional width in collapsed bowel segment.     

Probiotic characteristics and aflatoxin B1 binding ability of Debaryomyces hansenii and Kazaschtania exigua from rainbow trout environment

The aim of this study was to evaluate probiotic properties and the aflatoxin B₁ adsorption ability of yeasts isolated from rainbow trout intestine and fish feed to assess their use in the formulation of feed additives. Growth at pH 2, bacterial pathogens inhibition, bacterial pathogens co‐aggregation, autoaggregation, homologous and heterologous inhibition against lactic acid bacteria were evaluated. Moreover, aflatoxin B₁ adsorption was tested. All strains were able to maintain viable (10⁷ cells/ml) at pH 2. All strains isolated from intestine were identified as Kazaschtania exigua, while strains isolated from feed were all identified as Debaryomyces hansenii. Kazaschtania exigua RC035 and RC037 showed the strongest antimicrobial activity while K. exigua RC037 and RC038 were the most efficient co‐aggregating bacterial pathogens. All strains exhibited strong autoaggregation. None of the tested yeast strains showed homologous inhibition towards other yeasts and heterologous inhibition towards lactic acid bacteria strains. Debaryomyces hansenii RC031 demonstrated aflatoxin B₁ adsorption capacity (21%). The results of the present study indicate that select strains of Kazaschtania exigua and D. hansenii showed potential to improve the health of rainbow trout by inhibiting pathogens and binding AFB₁ and their use as probiotics may improve the production of rainbow trout in aquaculture systems.     

Pro-oxidant and antioxidant responses in the liver and kidney of wild Goodea gracilis and their relation with halomethanes bioactivation

In mammals, it has been shown that halomethanes (HM) are bioactivated by enzymes such as CYP 2E1 and the theta isoform of GST to produce reactive metabolites. However, in fish, little information is available, although HM can form autochthonously in aquatic environments. This study assessed the effect of HM in dusky splitfin (Goodea gracilis) from three lakes of the Valley of Mexico by analysing specific HM biomarkers as well as a broad range of biomarkers. The concentration of HM was a function of its half-life (higher in deep waters), while its precursors and solar radiation are secondary factors that determine its concentration. The kidney showed higher basal metabolism than the liver, probably because of its function as a haematopoietic and filtration organ. Using integrated biological response version 2 (IBRv2), it was found that the hepatic and renal O₂· content is a pro-oxidant force capable of inducing oxidative stress (ROOH, TBARS and RC=O). Early damage was found to be dependent on low concentrations of HM in Major Lake, whereas late damage was observed in fish exposed to higher concentrations of HM in Zumpango Lake and Ancient Lake. The activities of enzymes involved in antioxidant defence seemed to be inefficient. The quantitative assessment of biomarkers (ANOVA) and the estimate of parameter A obtained from IBRv2 provided different information. However, the data support the greater predictive power of IBRv2, but it requires a series of interrelated biomarkers to infer these possibilities. G. gracilis presents marked patterns of adaptation, which are dependant on the HM concentrations in environmental mixtures, although the response is complex and many toxicants could induce similar responses.     

Conserving seeds of useful wild plants in Mexico: main issues and recommendations

The efficient storage and germination of seeds underpin the effective use of plants for livelihoods and sustainable development. A total of 204 wild species useful for local communities of the Tehuacán–Cuicatlán Valley were collected and stored in seed banks in country for long term conservation, and 66 % (i.e., 134) duplicated in the U.K., as an effective means of ex situ conservation. Of the 204 species, 147 (122 of which also duplicated in the U.K.) were previously listed as useful plants in the ethnofloristic inventory of the Valley. Based on literature surveys, we found that one of the major impediments to the use of stored seeds of wild species is the lack of knowledge of how to germinate the seed. In detailed studies, we found that seeds of 18 useful plant species from 10 different families germinated readily and could be propagated. In contrast, four species (Actinocheita filicina, Bursera submoniliformis, Karwinskia mollis and Lippia graveolens) produced dormant seeds and therefore further studies are needed before their use can be maximised in large scale propagation programmes in support of conservation and livelihoods. Overall, this large-scale study on useful wild plant species in Mexico confirms that conventional seed banking can effectively support sustainable development and livelihood programmes.