Hepatitis E virus in Cuba: A cross-sectional serological and virological study in pigs and people occupationally exposed to pigs

Surveillance of hepatitis E virus (HEV) in risk groups is an important strategy to monitor its circulation pattern and to timely detect changes thereof. The aims of this cross-sectional study were to estimate the prevalence of HEV infections in pigs and humans from different regions of the country, to identify risk factors for increasing anti-HEV IgG prevalence and to characterize HEV strains. The presence of anti-HEV antibodies was assessed by commercial ELISA in serum samples from the general population, farm and slaughterhouse employees, as well as pigs sampled in the three regions of Cuba from February to September 2016. Overall, individuals with occupational exposure to swine or swine products (70/248, 28.2%) were 4 times more likely to be seropositive compared to the general population (25/285, 8.7%; OR: 4.18; p < .001). Within the risk group, risk factors included age, number of years working in a professional activity with direct exposure to swine, geographic region and distance between residence and closest professional swine setting, while wearing gloves had a protective effect. Prevalence of total anti-HEV antibodies in swine was 88.2% (165/187) and HEV RNA was detected by real-time RT-PCR in 9.2% (16/173) swine stools. All HEV strains sequenced clustered within genotype 3. Some strains clearly belonged to subtype 3a, while another group of strains was related with subtypes 3b and 3 k but partial HEV sequences did not allow unequivocal subtype assignment. These findings suggest that the high HEV exposure in Cuban individuals with swine-related occupations could be due to enzootic HEV in certain regions, direct contact with infectious animals or their products as well as environmental contamination.     

Effect of the addition of sewage sludge as a fertilizer on a sandy vineyard soil

Purpose

The application of sludge from wastewater in agriculture has increased in recent years, and it is therefore important to assess the effect that such treatment has on both the soil and the plant. The aim of the study described here was to ascertain whether there is a variation in the properties of the soil and to determine if this addition has an impact on the plant.

Materials and methods

The area of investigation was close to the municipality of Villarrubia de los Ojos (Ciudad Real). In this work, six samples were taken from the surface horizon in the studied plot at a depth of 35 cm. A further three samples were taken: (i) a surface horizon of a soil close to the area under investigation but without treatment (control sample), (ii) a sample of sludge from the wastewater treatment plant and (iii) a sample of the mixture used by farmers as fertilizer. Laboratory tests were conducted in accordance with the SCS-USDA (1972) guidelines. Trace element samples were analysed by X-ray fluorescence spectrophotometry (Philips PW 2404).

Results and discussion

The parcel of land studied is dominated by a sandy texture (88.3 % sand), and a decrease in pH was observed in areas in which the mixture (manure + sludge) was added (pH?=?8.0) compared to areas in which fertilizer was not applied (pH?=?8.5). It was observed that the addition of the compound led to an increase in the electrical conductivity of the soil. The trace elements can be organized into two groups based on the results obtained in this study. One group contains the trace elements that were only present in the rows that were treated with the fertilizer. The other group of trace elements was mobilized throughout the whole plot.

Conclusions

The application of sewage sludge on agricultural soils can be very useful as an organic amendment because it produces an increase in soil organic matter. However, sewage sludge must be applied with caution due to the changes in soil chemical properties (for example, pH and E.C.). The use of this type of waste for prolonged periods of time can cause problems of contamination in the soil.

     

Spray Drying of Chitosan Acid Salts: Process Development,Scaling Up and Physicochemical Material Characterization

We investigated a spray drying process for preparing water-soluble salts of high molecular weight chitosan (CH) intended for pharmaceutical excipient applications. CH was derived from chitin of marine lobster origin (Panulirus argus). The effects of organic acid (acetic or lactic acid) and the ratio (difference) of inlet/outlet air temperature (140/90 °C or 160/100 °C) on spray drying were studied. The yield of spray-dried CH salt powders ranged from 50% to 99% in laboratory and industrial-scale processes. The spray-dried dry powder of CH salts consisted of spherical agglomerated particles with an average diameter of 36.2 ± 7.0 µm (CH acetate) and 108.6 ± 11.5 µm (CH lactate). After dispersing the spray-dried CH salt powder samples in purified water, the mean particle sizes obtained for the CH acetate salts were 31.4 nm (batch A001), 33.0 nm (A002) and 44.2 nm (A003), and for the CH lactate salts 100.8 nm (batch L001), 103.2 nm (L002) and 121.8 nm (L003). The optimum process conditions for spray drying were found: an inlet air temperature of 160 ± 5 °C, an outlet temperature of 100 ± 5 °C and an atomizer disk rotational speed of 18,200 min⁻¹. The X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC) results confirmed the amorphous state of the CH salts. The ¹H nuclear magnetic resonance (NMR) and Fourier transform infrared (FT-IR) spectra of CH acetate and lactate salts verified that the spray drying process does not affect the polymer backbone. In conclusion, both laboratory and industrial-scale spray drying methods for preparing water-soluble acid salts of CH are reproducible, and the physicochemical properties of the corresponding CH acid salts are uniform.     

IL-17A regulates Eimeria tenella schizont maturation and migration in avian coccidiosis

Although IL17A is associated with the immunological control of various infectious diseases, its role in host response to Eimeria infections is not well understood. In an effort to better dissect the role of IL17A in host-pathogen interactions in avian coccidiosis, a neutralizing antibody (Ab) to chicken IL17A was used to counteract IL17A bioactivity in vivo. Chickens infected with Eimeria tenella and treated intravenously with IL17A Ab, exhibited reduced intracellular schizont and merozoite development, diminished lesion score, compared with untreated controls. Immunohistological evaluation of cecal lesions in the parasitized tissues indicated reduced migration and maturation of second-generation schizonts and reduced lesions in lamina propria and submucosa. In contrast, untreated and infected chickens had epithelial cells harboring second-generation schizonts, which extend into the submucosa through muscularis mucosa disruptions, maturing into second generation merozoites. Furthermore, IL17A Ab treatment was associated with increased parameters of Th1 immunity (IL2- and IFNγ- producing cells), reduced levels of reactive oxygen species (ROS), and diminished levels of serum matrix metalloproteinase-9 (MMP-9). Finally, schizonts from untreated and infected chickens expressed S100, Wiskott-Aldrich syndrome protein family member 3 (WASF3), and heat shock protein-70 (HSP70) proteins as merozoites matured, whereas the expression of these proteins was absent in IL17A Ab-treated chickens. These results provide the first evidence that the administration of an IL17A neutralizing Ab to E. tenella-infected chickens inhibits the migration of parasitized epithelial cells, markedly reduces the production of ROS and MMP-9, and decreases cecal lesions, suggesting that IL17A might be a potential therapeutic target for coccidiosis control.     

Variations in mercury and other trace elements contents in soil and in vine leaves from the Almadén Hg-mining district

Purpose

Vines (Vitis vinifera, L.) are a very important agricultural resource for Spain in general and for the Castilla-La Mancha region in particular, providing important productions of wines. Grapes and raisins are used for direct consumption too. In this work, we study analytical constraints regarding metallic trace elements uptake, focusing on Hg, from vines growing in the Almadén mercury mining district, the world's largest producer of this element, inactive nowadays.

Materials and methods

The study started with the analysis of these metals in soils and sets of vines leaves from seven sites located at different distances from the Almadén Hg mine. The samples of soils were dried at ambient temperature for 1 week. They were then sifted (<2 mm) and were stored for subsequent analysis. The leaves were dried and the leaf blade and petiole were separated. About 2 g of each sample were hand milled and analyzed using the same fluorescence spectrometer. Total mercury in soils and vine leaves were determined using a Lumex RA-915+ device, an atomic absorption spectrometer with a pyrolysis unit (RP-91c).

Results and discussion

Results show significant correlations between soil and leaves contents for total and organic mercury (R?=?0.934 and 0.984, respectively). Hg contents range in soil from 2,376 to 0.04 mg/kg in non-polluted places. For the organic fraction, the range varies between 197.49 and 3.15 mg/kg. Total Hg contents measured in leaves were from 5.14 mg/kg (close to dump zone of the mine) to 0.03 mg/kg in the proximity of Carrión de Calatrava, located some 100 km away from Almadén. Mercury reaches maximum in the proximity of known sources of the element: the mining and/or metallurgical areas of Almadén and Almadenejos.

Conclusions

Soils from the study area contain normal contents in trace metals, and these are conditioned by the local geology or urban locations of the area. Trace metals contents in leaves do not show a relationship with soil contents, possibly due to the low bioavailability of these elements in the soils investigated. Mercury is, as expected for this area, an exception to this trend, with very high concentrations that reach maximum values in the proximity of the known sources of the element: the proximity of mining and/or metallurgical areas in Almadén and Almadenejos.     

Bioaccumulation of mineral elements in grapevine varieties cultivated in “La Mancha”

The capacity of a plant to take up chemical elements is measured as the ratio of the element concentration in the plant and its concentration in the soil. This ratio is called the bioaccumulation index (BAC). The choice of rootstock and variety can differentially affect nutrient uptake of grapevine (Vitis vinifera L.). Generally, only slight differences can be observed in the accumulation of N, P, and Zn. In contrast, wide variations are found in the absorption of K, Ca, Mg, and Cl. While studies on plant nutrients and major elements have been carried out in vineyards, very few references concern the trace‐element concentrations in leaves. In the study described here, five varieties (Airén, Cencibel, Garnacha, Cabernet Sauvignon, and Chardonnay) grown on a Calcisol Petric or Typic Calcixerept (typical of La Mancha) were compared in terms of leaf trace‐element concentrations. A total of three samples (20 healthy and completely developed leaves from the middle part of the shoot) were collected from three different plants of the same variety. Leaves were dried, milled, and x‐ray fluorescence analysis was performed. It was found that the variety of grapevine influences the uptake of four of the major elements: Al (low in Garnacha, [0.4 ± 0.2] g kg^–1), Ca (high in Cencibel, [32.6 ± 8.1] g kg^–1), P (low in Airén, [0.9 ± 0.4] g kg^–1) and K, which showed significant differences in almost all of the varieties (from [4.7 ± 1.8] g kg^–1 in Cencibel to [8.0 ± 10.1] g kg^–1 in Chardonnay). No statistically significant varietal differences were found for trace elements. The only exception was La, which was present at higher levels in the variety Cencibel ([7.5 ± 0.3] mg kg^–1). This finding allows to assess the geochemical fingerprinting of calcareous soils regardless of the grapevine variety studied.     

MDN-0170, a New Napyradiomycin from Streptomyces sp. Strain CA-271078

A new napyradiomycin, MDN-0170 (1), was isolated from the culture broth of the marine-derived actinomycete strain CA-271078, together with three known related compounds identified as 4-dehydro-4a-dechloronapyradiomycin A1 (2), napyradiomycin A1 (3) and 3-chloro-6,8-dihydroxy-8-α-lapachone (4). The structure of the new compound was determined using a combination of spectroscopic techniques, including 1D and 2D NMR and electrospray-time of flight mass spectrometry (ESI-TOF MS). The relative configuration of compound 1, which contains two independent stereoclusters, has been established by molecular modelling in combination with nOe and coupling constant analyses. Biosynthetic arguments also allowed us to propose its absolute stereochemistry. The antimicrobial properties of the compounds isolated were evaluated against methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli, Aspergillus fumigatus, and Candida albicans. The potent bioactivity previously reported for compounds 2 and 3 against methicillin-sensitive S. aureus has been extended to methicillin-resistant strains in this report.     

Feeding of tropical trees and shrub foliages as a strategy to reduce ruminal methanogenesis: studies conducted in Cuba

The aim of this paper was to present the main results obtained in Cuba on the effects of feeding tropical trees and shrubs on rumen methanogenesis in animals fed with low quality fibrous diets. More than 20 tree and shrub foliages were screened for phytochemicals and analyzed for chemical constituents. From these samples, seven promising plants (Samanea saman, Albizia lebbeck, Tithonia diversifolia, Leucaena leucocephala, Trichantera gigantea, Sapindus saponaria, and Morus alba) were evaluated for methane reduction using an in vitro rumen fermentation system. Results indicated that the inclusion levels of 25% of Sapindo, Morus, or Trichantera foliages in the foliages/grass mixtures (grass being Pennisetum purpureum) reduced (P < 0.01) methane production in vitro when compared to Pennisetum alone (17.0, 19.1, and 18.0 versus 26.2 mL CH₄/g fermented dry matter, respectively). It was demonstrated that S. saman, A. lebbeck, or T. diversifolia accession 23 foliages when mixed at the rate of 30% in Cynodon nlemfuensis grass produced lower methane compared to the grass alone. Inclusion levels of 15% and 25% of a ruminal activator supplement containing 29% of L. leucocehala foliage meal reduced methane by 37% and 42% when compared to the treatment without supplementation. In vivo experiment with sheep showed that inclusion of 27% of L. leucocephala in the diet increased the DM intake but did not show significant difference in methane production compared to control diet without this foliage. The results of these experiments suggest that the feeding of tropical tree and shrub foliages could be an attractive strategy for reduction of ruminal methanogenesis from animals fed with low-quality forage diets and for improving their productivity.