Genomic Description of Antibiotic Resistance in Escherichia coli and Enterococci Isolates from Healthy Lusitano Horses

Antibiotic resistance is a global problem, and it is known that commensal bacteria can act as reservoir of antibiotic resistance genes of clinical importance. The aim of the present study was to determine the antibiotic resistance phenotype and mechanisms implicated in resistance of Escherichia coli and Enterococcus spp. isolates collected from fecal samples of 90 Lusitano horses from Portugal. Sixteen of the 71 E. coli isolates (22.5%) recovered showed resistance to at least one of the antibiotics tested. The number of E. coli isolates resistant to streptomycin, tetracycline, chloramphenicol, ampicillin, trimethoprim-sulfamethoxazole, and gentamicin was 9, 7, 6, 3, 2, and 1, respectively. The bla_TEM-1 and bla_OXA-1 genes were detected in ampicillin-resistant isolates and the sul2 and dfrA1 genes in trimethoprim-sulfamethoxazole-resistant, while the aac(3)-I, floR and tet(A) were found in the gentamicin, chloramphenicol and tetracycline-resistant isolates, respectively. Twenty-two of the 71 (31%) recovered enterococci showed antibiotic resistance for at least one of the tested antibiotics, and resistant isolates were identified as Enterococcus faecium (n = 14), E. faecalis (n = 3), E. hirae (n = 2), and Enterococcus spp. (n = 3). The erm(B) and erm(C) genes were identified in erythromycin-resistant enterococci and the tet(M) and/or tet(L) genes in tetracycline-resistant isolates. The slight prevalence of antibiotic resistance among commensal bacteria of healthy Lusitano horses can improve the treatment of upcoming infections in these horses because these microorganisms can be considered as antimicrobial indicator bacteria.     

The Correlation between Age,Body Weight and Testicular Parameters in Murrah Buffalo Bulls Raised in Brazil

Buffalo are an economically important source for meat and milk production, especially in Brazil. However, important aspects of their biology remain unknown thus far. Herein, we describe the reproductive characteristics of male Murrah buffalo (Bubalus bubalis) raised under extensive management conditions by applying biometrics associated with testicular weight. We analyzed seven males, divided into two groups: G1, which consisted of four 18-month-old animals, and G2, which consisted of three 24-month-old animals. Testicular development occurs slowly in Murrah buffalo, suggesting a delay of sexual maturity. The biometric testicular parameters analyzed were scrotal circumference, testicular weight, testicular length, testicular width, testicular thickness and testicular circumference. Our data indicate strong correlations between SC, age and body weight, and additional significant relationships were identified between body weight, age and other testicular parameters. Thus, these parameters are suitable indicators when selecting bulls for breeding purposes.     

Chickens treated with a nitric oxide inhibitor became more resistant to Plasmodium gallinaceum infection due to reduced anemia,thrombocytopenia and inflammation

Malaria is a serious infectious disease caused by parasites of the Plasmodium genus that affect different vertebrate hosts. Severe malaria leads to host death and involves different pathophysiological phenomena such as anemia, thrombocytopenia and inflammation. Nitric oxide (NO) is an important effector molecule in this disease, but little is known about its role in avian malaria models. Plasmodium gallinaceum- infected chickens were treated with aminoguanidine (AG), an inhibitor of inducible nitric oxide synthase, to observe the role of NO in the pathogenesis of this avian model. AG increased the survival of chickens, but also induced higher parasitemia. Treated chickens demonstrated reduced anemia and thrombocytopenia. Moreover, erythrocytes at different stages of maturation, heterophils, monocytes and thrombocytes were infected by Plasmodium gallinaceum and animals presented a generalized leucopenia. Activated leukocytes and thrombocytes with elongated double nuclei were observed in chickens with higher parasitemia; however, eosinophils were not involved in the infection. AG reduced levels of hemozoin in the spleen and liver, indicating lower inflammation. Taken together, the results suggest that AG reduced anemia, thrombocytopenia and inflammation, explaining the greater survival rate of the treated chickens.     

Comparison of Different Glycerol and Egg Yolk Concentrations Added to Tris‐based Extender for the Collared Peccaries (Tayassu tajacu) Semen Freezing

This study aimed to evaluate various concentrations of egg yolk (5, 10, or 20%) in combination with different concentrations of glycerol (3% or 6%) added to a Tris‐based extender on the post‐thaw characteristics of sperm obtained from Tayassu tajacu. For this purpose, semen from 10 sexually male mature collared peccaries was collected by electroejaculation and evaluated for sperm motility, vigour, viability, morphology and functional membrane integrity. The ejaculates were initially extended in Tris‐fructose plus egg yolk (5%, 10% or 20%). After cooling, the semen was added to Tris‐egg yolk plus glycerol (6% or 12%), resulting in a final concentration of 3% or 6% glycerol of the extender. Straws were frozen using liquid nitrogen and thawed in a water bath at 37°C for 30 s. The frozen–thawed semen was evaluated as reported for fresh semen. After thawing, a significant decrease was verified for sperm motility and vigour, for all the samples in comparison with fresh semen. However, no differences were evidenced among treatments for any sperm characteristics evaluated (p > 0.05), except for the combination between 10% egg yolk and 6% glycerol, which provided the worst preservation of functional membrane integrity (p < 0.05). The interactions between higher concentrations of egg yolk (20%) and glycerol (6%) and also between lower concentrations of the same substances (5% egg yolk and 3% glycerol) added to the Tris‐based extender negatively affected the preservation of the normal sperm morphology after thawing (p < 0.05). In conclusion, the use of Tris‐based extender added to 10% or 20% egg yolk plus 3% glycerol is recommended for effective sperm cryopreservation in collared peccaries.     

Apparent electrical conductivity in dry versus wet soil conditions in a shallow soil

The general objective of this study was to evaluate the stability of patterns of apparent soil electrical conductivity (EC_a) in dry versus wet soil conditions in a shallow soil typically used for pastures in Mediterranean conditions of the southern region of Portugal. A 6 ha experimental field of permanent bio-diverse pasture was divided into 76 squares of 28 × 28 m. The soil electrical conductivity was measured using a Dualem 1S sensor under dry conditions (June 2007) and under wet conditions during the rainy season (March 2010). Soil samples, geo-referenced with GPS, were collected in a depth range of 0–0.30 m. The soil was characterized in terms of bedrock depth, moisture content, texture, pH, organic matter content, and macronutrients (nitrogen, phosphorus, and potassium). Pasture samples, also geo-referenced with GPS, were collected to measure the pasture dry matter yield. The statistical analysis of apparent electrical conductivity between dry and wet soil conditions resulted in a linear significant correlation coefficient (R = 0.88). The results also showed a significant correlation between apparent electrical conductivity and the relative field elevation (R = ?0.64 and R = ?0.66), the pasture dry matter yield (R = 0.42 and R = 0.48), the bedrock depth (R = 0.40 and R = 0.27), the pH (R = 0.50 and R = 0.49), the silt (R = 0.27 and R = 0.38) and soil moisture content (R = 0.48 and R = 0.45), in dry and wet conditions, respectively. A multi-variate regression was carried out using the following soil parameters that showed significant correlation with EC_a and that did not present multi-collinearity: pH, bedrock depth, silt and moisture content. The results showed, in dry and wet conditions, that the analysis was significant (R = 0.75 and R = 0.84, respectively). Overall, these results indicate the temporal stability of EC_a patterns under different soil moisture contents, which is relevant with respect to the time when a field should be surveyed and is important for using the electrical conductivity sensor, as a decision support tool for management zones in precision agriculture.     

Performance of ‘Coscia’ pear (Pyrus communis) on seven rootstocks in a warm climate

Summary

The vegetative and reproductive performance of ‘Coscia’ pear (Pyrus communis L.) growing on seven rootstocks [OHF 69, OHF 97, OHF 513 and BP 1 (P. communis), clonal seedling (Davis AxB) of P. betulifolia, and quince BA 29 and EMA (Cydonia oblonga)] were compared over an 8-year period. The trial was conducted at the Experimental Orchard Farm Station in northern Israel, on a well-drained soil with pH 7.5. Trees were planted in December 1998 at a distance of 4.0 m 2.0 m, and trained with a central axis. The most vigorous trees were on P. betulifolia seedling, followed by BP 1 and the three OHF rootstocks (69, 97, 513). All the above rootstocks demonstrated greater vigour than quince BA 29 or EMA. The reason for this effect, at least in part, appeared to be the excellent water status (high midday stem water potential values) of trees on P. betulifolia in comparison with the other rootstocks. The highest cumulative yields per tree were harvested from trees on P. betulifolia and BP 1, followed by the three OHF rootstocks (69, 97, 513). However, the highest cumulative yield of large fruit (> 60 mm) was obtained from trees on P. betulifolia, followed by the OHF series and BP 1. The two quince rootstocks had the lowest cumulative yield, and the lowest yield of large fruit. A positive correlation was found between the vigour of the tree, as affected by the rootstock, and both total yield and fruit size. We conclude that, in a warm climate, yield efficiency is not the only parameter that should be taken into account, and building a strong tree for the weak scion cultivar is the first requirement for establishing an orchard. Fruit quality at harvest and during cold storage were examined for fruit from three rootstocks only. The highest soluble solids content values at harvest were obtained in fruit grown on quince EMA, compared to values for BP 1 and P. betulifolia fruit.     

Evaluating ecological‐niche factor analysis as a modelling tool for environmental weed management in island systems

Management actions are essential for mitigating the potentially harmful changes in biodiversity, ecosystem function and crop/forest productivity caused by invasive species. Species distribution models, if reliable, could be used to design effective management strategies. Although several modelling methods well suited for studying invasive species have been developed for presence‐only data, often the size of available data sets for modelling is small and results are not validated with test samples. Moreover, the impact of such methods in practical applications has been overlooked. Here, we evaluated the reliability of the modelling approach based on ecological‐niche factor analysis (ENFA) implemented in Biomapper software when applied to environmental weed data in the Azores. Presence‐only data sets of two top invasive woody species (Pittosporum undulatum and Acacia melanoxylon) were used. The continuous Boyce curve was used for validation, calculated either in Biomapper (cross‐validation) or based on test samples. The species' most habitable areas that should be regarded as management targets were thus estimated from modelling and validation. By imposing size restrictions on the presence‐only data sets used in modelling and validation, other habitable areas were defined and compared. The ENFA proved to be a suitable method for modelling environmental weed distributions, regardless of the presence‐only dataset size. Moreover, the cross‐validation of Biomapper was reliable, although its results should be interpreted with caution as they could potentially lead to statistically different management target areas.     

Development and validation of a set of standard area diagrams to estimate severity of potato early blight

Standard area diagrams (SADs) to assess the severity of potato early blight (Alternaria grandis) on leaves of potato (Solanum tuberosum L.) were developed and validated. The proposed SADs include images of leaves with 10 distinct disease severities (0.1, 1, 3, 5, 10, 20, 40, 60, 80 and 100 %). The SADs were validated by 12 raters who had no previous experience in evaluating plant disease. Lin’s concordance correlation analysis of estimated vs. actual disease severity (based on image analysis) showed that precision and accuracy improved for most raters using the SADs, compared to assessments made without the SADs. The SADs improved accuracy (coefficient of bias, C _b ?=?0.97 and 0.99, without and with SADs, respectively) and agreement (Lin’s concordance correlation coefficient, ρ _c ?=?0.91 and 0.98 without and with SADs, respectively) of the estimates of severity. Severity estimates were more reliable when using SADs (coefficient of determination, R ² ?=?0.80 unaided and R ² ?=?0.95 with SADs, and the intra-class correlation ρ?=?0.86 without SADs and ρ?=?0.97 using the SADs). The SADs improved raters’ ability to accurately, precisely and reliably estimate potato early blight severity, and as such can be used to assess severity for several purposes, including breeding for resistance, fungicide screening, and pathotype characterization.     

Brassica oleracea L. Var. costata DC and Pieris brassicae L. aqueous extracts reduce methyl methanesulfonate-induced DNA damage in V79 hamster lung fibroblasts

Brassica oleracea L. var. costata DC leaves and Pieris brassicae L. larvae aqueous extracts were assayed for their potential to prevent/induce DNA damage. None of them was mutagenic at the tested concentrations in the Ames test reversion assay using Salmonella His(+) TA98 strains, with and without metabolic activation. In the hypoxanthine-guanine phosphoribosyltransferase mutation assay using mammalian V79 fibroblast cell line, extracts at 500 μg/mL neither induced mutations nor protected against the mutagenicity caused by methyl methanesulfonate (MMS). In the comet assay, none of the extracts revealed to be genotoxic by itself, and both afforded protection, more pronounced for larvae extracts, against MMS-induced genotoxicity. As genotoxic/antigenotoxic effects of Brassica vegetables are commonly attributed to isothiocyanates, the extracts were screened for these compounds by headspace-solid-phase microextraction/gas chromatography-mass spectrometry. No sulfur compound was detected. These findings demonstrate that both extracts could be useful against damage caused by genotoxic compounds, the larvae extract being the most promising.