Quantification of transmission of foot-and-mouth disease virus caused by an environment contaminated with secretions and excretions from infected calves

Foot-and-mouth disease virus (FMDV) infected animals can contaminate the environment with their secretions and excretions. To quantify the contribution of a contaminated environment to the transmission of FMDV, this study used calves that were not vaccinated and calves that were vaccinated 1 week prior to inoculation with the virus in direct and indirect contact experiments. In direct contact experiments, contact calves were exposed to inoculated calves in the same room. In indirect contact experiments, contact calves were housed in rooms that previously had held inoculated calves for three days (either from 0 to 3 or from 3 to 6 days post inoculation). Secretions and excretions from all calves were tested for the presence of FMDV by virus isolation; the results were used to quantify FMDV transmission. This was done using a generalized linear model based on a 2 route (2R, i.e. direct contact and environment) SIR model that included information on FMDV survival in the environment. The study shows that roughly 44% of transmission occurs via the environment, as indicated by the reproduction ratio R^02Renvironment that equalled 2.0, whereas the sum of R^02Rcontact and R^02Renvironment equalled 4.6. Because vaccination 1 week prior to inoculation of the calves conferred protective immunity against FMDV infection, no transmission rate parameters could be estimated from the experiments with vaccinated calves. We conclude that a contaminated environment contributes considerably to the transmission of FMDV therefore that hygiene measures can play a crucial role in FMD control.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-015-0156-5) contains supplementary material, which is available to authorized users.     

Immunization with Eimeria ninakohlyakimovae-live attenuated oocysts protect goat kids from clinical coccidiosis

Caprine coccidiosis, affecting mainly young goat kids around the weaning period, is worldwide the most important disease in the goat industry. Control of caprine coccidiosis is increasingly hampered by resistances developed against coccidiostatic drugs leading to an enhanced need for anticoccidial vaccines. In the current study we conducted an oral immunization trial with live attenuated sporulated Eimeria ninakohlyakimovae oocysts. Sporulated E. ninakohlyakimovae oocysts were attenuated by X-irradiation technique. The experimental design included a total of 18 goat kids divided into the following groups: (i) animals immunized with attenuated E. ninakohlyakimovae oocysts at 5 weeks of age and challenged 3 weeks later with non-irradiated homologous oocysts (group 1); (ii) animals infected with non-attenuated E. ninakohlyakimovae oocysts at 5 weeks of age and challenged 3 weeks later with non-attenuated homologous oocysts (group 2); (iii) animals primary-infected with untreated E. ninakohlyakimovae oocysts at 8 weeks of age (control of the challenge infection, group 3); (iv) non-infected control animals (group 4). Goat kids immunized with live attenuated E. ninakohlyakimovae oocysts (group 1) excreted significantly less oocysts in the faeces (95.3% reduction) than kids infected with non-attenuated ones (group 2). Furthermore, immunization with live but attenuated oocysts resulted in ameliorated clinical coccidiosis compared to goat kids infected with untreated oocysts (group 2) and resulted in equally reduced signs of coccidiosis after challenge infection compared to acquired immunity driven by non-attenuated oocysts. Overall, the present study demonstrates for the first time that live attenuated E. ninakohlyakimovae oocysts orally administered showed almost no pathogenicity but enough immunogenicity in terms of immunoprotection. Importantly, vaccinated animals still shed low amounts of oocysts, guaranteeing environmental contamination and consecutive booster infections to sustain ongoing immunity.     

EcPV2 DNA in equine genital squamous cell carcinomas and normal genital mucosa

Squamous cell carcinoma (SCC) represents the most common genital malignant tumor in horses. Similar to humans, papillomaviruses (PVs) have been proposed as etiological agents and recently Equine papillomavirus type 2 (EcPV2) has been identified in a subset of genital SCCs. The goals of this study were (1) to determine the prevalence of EcPV2 DNA in tissue samples from equine genital SCCs, penile intraepithelial neoplasia (PIN) and penile papillomas, using EcPV2-specific PCR, (2) to examine the prevalence of latent EcPV2 infection in healthy genital mucosa and (3) to determine genetic variability within EcPV2 and to disentangle phylogenetic relationships of EcPV2 among PVs. EcPV2 DNA was detected in all but one penile SCC (15/16), in all PIN lesions (8/8) and penile papillomas (4/4). Additionally, EcPV2 DNA was demonstrated in one of two metastasized lymph nodes, one contact metastasis in the mouth, two vaginal and one anal lesion. In healthy horses, EcPV2 DNA was detected in 10% (4/39) of penile swabs but in none of vulvovaginal swabs (0/20). This study confirms the presence of EcPV2 DNA in equine genital SCCs and shows its involvement in anal lesions, a lymph node and contact metastases. Latent EcPV2 presence was also shown in normal male genital mucosa. We found that different EcPV2 variants cocirculate among horses and that EcPV2 is related to the Delta+Zeta PVs and is only a very distant relative of high-risk human PVs causing genital cancer. Thus, similar viral tropism and similar malignant outcome of the infection do not imply close evolutionary relationship.     

Evaluating potassium phosphonate injections for the control of <Emphasis Type="Italic">Quercus ilex</Emphasis> decline in SW Spain: implications of low soil contamination by <Emphasis Type="Italic">Phytophthora cinnamomi</Emphasis> and low soil water content on the effectiveness of treatments

The Iberian forests are suffering severe disease and mortality as a result of decline, with Quercus ilex the major species at risk. Trunk injections with potassium phosphonate, which have been used successfully to control Phytophthora cinnamomi, were tested against decline. In an area in which P. cinnamomi was isolated, Q. ilex trees showing different degrees of decline were trunk-injected. Soil properties, and measurements of soil water content (θ) and depth to soil water table were assessed at three sites with markedly different decline incidences. Over the 5 years following the initiation of the experiment, mean symptoms among spring-treated trees and autumn-treated trees, or among trees injected twice a year (spring and autumn), once a year, and non-injected, were not significantly different. No effects of the treatments on shoot growth and acorn production were observed. However, θ values under trees which recovered from decline were higher than θ values under trees which did not recover from decline. At the site with the highest incidence of decline and tree mortality, P. cinnamomi was rarely isolated, and the presence of gravel, soil infiltration capacities and water table depth values were significantly higher than at the other sites, water stress being more likely to contribute to decline than P. cinnamomi. In areas in which θ is low, the distribution of phosphonate on the tree would be limited. Since the thresholds for phytotoxicity of potassium phosphonate in Q. ilex trees at the site studied would be higher than the amounts used, rates of the chemical slightly less than those that cause phytotoxicity should be tested.     

Pacific approaches to fundraising in the digital age: COVID-19, resilience and community relational economic practices

The aim of this paper is to discuss how community relational economic practices in virtual spaces are effective in building resilience because they are borne of and sustained by familiar traditional Fijian values of collective work and social interdependence. The researchers adopted a pandemic-induced methodology, conducting online-based talanoa (fluid conversations between two or more people) with a number of people leading, or involved in, these initiatives. We also engaged with online community groups behind a number of initiatives. Examples are provided of online crowdfunding, livestreaming of concerts to solicit donations, and bartering facilitated by social media sites. To conclude, we stress the enduring nature of communal bonds and traditional systems which Pacific people readily adapt and translate into different forums and forms in the face of challenges such as the restrictions and financial hardships caused by COVID-19. The findings highlight that solesolevaki – a tradition of working together for a common cause – can also occur in the digital era: this demonstrates the deep connection of Fijian peoples and their sense of obligation to one another and to their culture, regardless of where they are in the world.     

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.     

Role of lysine epsilon-amino groups of beta-lactoglobulin on its activating effect of Kluyveromyces lactis beta-galactosidase

Native beta-lactoglobulin binds and increases the activity of Kluyveromyces lactis beta-galactosidase. Construction of a three-dimensional (3D) model of beta-lactoglobulin showed that lysine residues 15, 47, 69, and 138 are the most exposed ones, thus the ones more likely to interact with beta-galactosidase. Molecular docking estimated the interaction energies of amino acid residues with either lactose or succinic anhydride, showing that Lys(138) is the most likely to react with both. Affinity chromatography demonstrated that succinylated beta-lactoglobulin diminished its ability to bind to the enzyme. Furthermore, when activity was measured in the presence of succinylated beta-lactoglobulin, its activating effect was lost. Since succinylation specifically blocks Lys epsilon-amino groups, their loss very likely causes the disappearance of the activating effect. Results show that the activating effect of beta-lactoglobulin on beta-galactosidase activity is due to the interaction between both proteins and that this interaction is very likely to occur through the Lys epsilon-amino groups of beta-lactoglobulin.     

Determination of the secondary structure of Kluyveromyces lactis beta-galactosidase by circular dichroism and its structure-activity relationship as a function of the pH

The secondary structure of Kluyveromyces lactis beta-galactosidase was determined by circular dichroism. It is mainly a beta-type protein, having 22% beta-turns, 14% parallel beta-sheet, 25% antiparallel beta-sheet, 34% unordered structure, and only 5% alpha-helix. The structure-activity relationship as a function of the pH was also studied. The pH conditions leading to the highest secondary structure content (100% ellipticity) of the enzyme was found at pH 7.0; at pH 6.5-7.0, the percent ellipticity decreased slightly, suggesting little structural change, but the activity decreased significantly, probably because of variations in critical residues. On the other hand, at pH's above 7.0, a more noticeable change in ellipticity was observed due to structural changes; the CD analysis showed a small increase in the helical content toward higher pH, whereas the maximum activity was found at pH 7.5, meaning that the changes produced in the secondary structure at this pH favored the interaction between the enzyme and the substrate.     

Changes in water retention properties due to the application of sugar foam in red soils

This work describes the influence of the application of sugar foam (an organic residue from sugar beet industry) on water retention properties, over an extended period of time (>25 years), in two red soils in the La Mancha region of Spain. The properties of gravimetric moisture at field capacity, gravimetric moisture in the permanent wilting point and available water retention capacity both in the original soil - without the addition of sugar foam - and in the soil affected by the addition of sugar foam are compared. For this purpose, the profiles are characterised macro morphologically. Chemical, physical-chemical and mineralogical parameters are determined, in addition to determining the water retention curves of each soil with tensiometers and Richards pressure plates.The sugar foam applied to the soil altered its physical and chemical properties. An increase in contents of organic matter (3.5% versus 1.4%), calcium carbonate (40.8% versus 0%) and pH (8.2 versus 6.3) are observed in the superficial horizon of the studied soils, although there is no such significant increase in electrical conductivity (0.33 dS/m versus 0.25 dS/m). With regard to the physical properties, the depth of horizon A increases (32 cm versus 12 cm), the stoniness reduces (5% versus 25%), the structure is well developed, due to the texture becomes finer (silty versus sandy clay loam), and, finally, the bulk density reduces (0.79 g cm⁻³ versus 1.19 g cm⁻³).The gravimetric moisture at field capacity was 49% in the soil altered with sugar foam, versus 12% in the non-altered soil; the gravimetric moisture in the permanent wilting point was 14.5% versus 8% and the available water retention capacity was 34.5% and 4%, respectively. It can be concluded that the increase of calcium carbonate, organic matter and the reduction of bulk density are the most influential factors in this process. The scientific novelty of this work is that the hydro-behaviour of the soil due to the addition of sugar foam improves the characteristic values of moisture and, therefore, the agronomical qualities of the soil.     

Comparison of two analytical methods for the evaluation of the complexed metal in fertilizers and the complexing capacity of complexing agents

The aim of this research is to develop an analytical methodology for the determination of complexed element in fertilizers and, then, to obtain an adequate criteria for the inclusion of these products in European Regulations on Fertilizers. This paper compares the CEN method EN 13366:2001, based on the retention of the cations into a sulfonated resin, and an AOAC modified method, based on the precipitation of the inorganic forms at pH 9. A limited interlaboratory trial was carried out to demonstrate the applicability of the AOAC modified method and to study the effect of the removal of organic compounds and the addition of a matrix modifier solution before the element quantification. Then, a global interlaboratory trial was developed to evaluate the validation and quality parameters of the method. As a second objective, the AOAC modified method was applied to the determination of the complexing capacity of complexing agents based on lignosulfonates and amino acids. The AOAC modified method was the choice methodology because it is adequate for the determination of complexing capacity of micronutrients in fertilizer.