Overproduction of the rhizobial adhesin RapA1 increases competitiveness for nodulation

Competition for nodulation is a complex problem where bacterial adhesins, which are required for root colonization, may play a role. However, the possible influence of adhesins on competitiveness was scarcely studied. In this work, the Rhizobium leguminosarum bv trifolii adhesion protein RapA1 was overproduced from a pHC60-derived plasmid and expressed in R200 strain. When an overproducing strain and a control-carrying empty vector were co-inoculated on clover plants, a positive effect of RapA1 on competition for nodule occupation was observed. Therefore, optimization of RapA1 expression may be considered for improvement of rhizobial competitiveness.     

Management of a caseous lymphadenitis outbreak in a new Iberian ibex (Capra pyrenaica) stock reservoir

Background

In 2010, an Iberian ibex (Capra pyrenaica hispanica) stock reservoir was established for conservation purposes in north-eastern Spain. Eighteen ibexes were captured in the wild and housed in a 17 hectare enclosure. Once in captivity, a caseous lymphadenitis (CLA) outbreak occurred and ibex handlings were carried out at six-month intervals between 2010 and 2013 to perform health examinations and sampling. Treatment with a bacterin-based autovaccine and penicillin G benzatine was added during the third and subsequent handlings, when infection by Corynebacterium pseudotuberculosis was confirmed. Changes in lesion score, serum anti-C. pseudotuberculosis antibodies and haematological parameters were analyzed to assess captivity effects, disease emergence and treatment efficacy. Serum acute phase proteins (APP) Haptoglobin (Hp), Amyloid A (SAA) and Acid Soluble Glycoprotein (ASG) concentrations were also determined to evaluate their usefulness as indicators of clinical status.Once in captivity, 12 out of 14 ibexes (85.7%) seroconverted, preceding the emergence of clinical signs; moreover, TP, WBC, eosinophil and platelet cell counts increased while monocyte and basophil cell counts decreased. After treatment, casualties and fistulas disappeared and both packed cell volume (PCV) and haemoglobin concentration significantly increased. Hp, SAA and ASG values were under the limit of detection or showed no significant differences.

Conclusions

A role for captivity in contagion rate is suggested by the increase in antibody levels against C. pseudotuberculosis and the emergence of clinical signs. Although boosted by captivity, this is the first report of an outbreak of caseous lymphadenitis displaying high morbidity and mortality in wild ungulates. Treatment consisting of both vaccination and antibiotic therapy seemed to prevent mortality and alleviate disease severity, but was not reflected in the humoural response. Haematology and APP were not useful indicators in our study, perhaps due to the sampling frequency. Presumably endemic and irrelevant in the wild, this common disease of domestic small ruminants is complicating conservation efforts for the Iberian ibex in north-eastern Spain.     

Vineyard yield estimation by combining remote sensing,computer vision and artificial neural network techniques

In viticulture, it is critical to predict productivity levels of the different vineyard zones to undertake appropriate cropping practices. To overcome this challenge, the final yield was predicted by combining vegetation indices (VIs) to sense the health status of the crop and by computer vision to obtain the vegetated fraction cover (F_c) as a measure of plant vigour. Multispectral imagery obtained from an unmanned aerial vehicle (UAV) is used to obtain VIs and F_c, which are used together with artificial neural networks (ANN) to model the relationship between VIs, F_c and yield. The proposed methodology was applied in a vineyard, where different irrigation and fertilisation doses were applied. The results showed that using computer vision techniques to differentiate between canopy and soil is necessary in precision viticulture to obtain accurate results. In addition, the combination of VIs (reflectance approach) and F_c (geometric approach) to predict vineyard yield results in higher accuracy (root mean square error (RMSE)?=?0.9 kg vine^?1 and relative error (RE)?=?21.8% for the image when close to harvest) compared to the simple use of VIs (RMSE?=?1.2 kg vine^?1 and RE?=?28.7%). The implementation of machine learning techniques resulted in much more accurate results than linear models (RMSE?=?0.5 kg vine^?1 and RE?=?12.1%). More precise yield predictions were obtained when images were taken close to the harvest date, although promising results were obtained at earlier stages. Given the perennial nature of grapevines and the multiple environmental and endogenous factors determining yield, seasonal calibration for yield prediction is required.

     

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.     

Atlantic Salmon (Salmo salar L.) as a Marine Functional Source of Gamma-Tocopherol

Gamma tocopherol (gT) exhibits beneficial cardiovascular effects partly due to its anti-inflammatory activity. Important sources of gT are vegetable oils. However, little is known to what extent gT can be transferred into marine animal species such as Atlantic salmon by feeding. Therefore, in this study we have investigated the transfer of dietary gT into salmon. To this end, fish were fed a diet supplemented with 170 ppm gT for 16 weeks whereby alpha tocopherol levels were adjusted to 190 ppm in this and the control diet. Feeding gT-rich diets resulted in a three-fold increase in gT concentrations in the liver and fillet compared to non-gT-supplemented controls. Tissue alpha tocopherol levels were not decreased indicating no antagonistic interaction between gamma- and alpha tocopherol in salmon. The concentration of total omega 3 fatty acids slightly increased in response to dietary gT. Furthermore, dietary gT significantly decreased malondialdehyde in the fillet, determined as a biomarker of lipid peroxidation. In the liver of gT fed salmon we observed an overall down-regulation of genes involved in lipid homeostasis. Additionally, gT improved the antioxidant capacity by up-regulating Gpx4a gene expression in the pyloric caeca. We suggest that Atlantic salmon may provide a marine functional source capable of enriching gT for human consumption.     

Serum concentrations of calcium, phosphorus, magnesium and calciotropic hormones in donkeys

OBJECTIVE: To provide reference values for serum biochemical variables that are used for evaluation of mineral metabolism in donkeys and compare values with those in horses. ANIMALS: 18 donkeys and 18 horses. PROCEDURES: Total calcium (tCa), total magnesium (tMg), and inorganic phosphorus (P) concentrations were measured in serum samples via spectrophotometry. Ionized calcium (iCa) and magnesium (iMg) concentrations were quantified with selective electrodes. By use of a micropartition system, tCa and tMg were fractionated to separate protein-bound (pCa, pMg) and ultrafiltrable fractions. Complexed calcium (cCa) and magnesium (cMg) concentrations were calculated by substracting ionized fractions from ultrafiltrable fractions. Parathyroid hormone (PTH) and calcitriol (CTR) concentrations were measured via radioimmunoassay. RESULTS: Serum tCa concentration in donkeys (3.37 +/- 0.21 mmol/L) was composed of pCa (1.59 +/- 0.21 mmol/L [47.0 +/- 4.2%]), iCa (1.69 +/- 0.04 mmol/L [50.4 +/- 3.0%]), and cCa (0.09 +/- 0.08 mmol/L [2.6 +/- 2.9%]). Serum tMg concentration (1.00 +/- 0.08 mmol/L) was fractioned in pMg (0.23 +/- 0.08 mmol/L [23.4 +/- 8.1%]), iMg (0.59 +/- 0.04 mmol/L [58.8 +/- 5.1%]), and cMg (0.18 +/- 0.08 mmol/L [17.8 +/- 7.2%]). Serum concentrations of P (1.14 +/- 0.30 mmol/L), PTH (20.4 +/- 21.2 pg/mL), and CTR (13.4 +/- 5.9 pg/mL) were determined. CONCLUSIONS AND CLINICAL RELEVANCE: Serum variables of mineral metabolism in donkeys were within reference ranges for horses. However, when compared with horses, donkeys had higher iCa, cMg, and CTR and lower pMg and PTH concentrations.     

Use of intraoperative impression smear cytology to guide successful treatment of a large renal cyst in a dog: a case report

A 13-yr-old Shih tzu was referred for surgical management of right-sided cranial abdominal mass, which corresponded to large, cavitated renal mass on ultrasonography, and was suspected to represent neoplasia. Intraoperative impression smear cytology (ISC) of the renal mass wall was consistent with benign renal cyst (RC), without evidence of neoplasia or infection. Deroofing and omentalisation were performed and histopathology was consistent with benign RC. Chronic kidney disease was diagnosed 4 mon postoperatively, however, the dog was asymptomatic, without cyst reoccurrence. Intraoperative ISC is an expedient and inexpensive diagnostic technique that can guide most appropriate treatment in dogs with large RCs.     

Assessment of cold tolerance at early developmental stages and allelic variation at candidate genes in South American rice germplasm

Rice is susceptible to cold during early developmental stages. Most tolerant cultivars have been developed for other conditions than those occurring in South America’s rice growing areas, or their grain type is not suitable for the local markets. If locally adapted varieties were available, growers could anticipate sowing date, making flowering time coincide with the moment of maximum solar radiation and increasing yields. In this work, 116 rice inbred lines and varieties of diverse origin within the South American gene pool were tested for seedling survival and germination percentage under low temperature in controlled conditions. As a result, lines used as controls responded as expected, whereas lines with similar behavior to controls, intermediate between susceptible and tolerant controls and more extreme than controls were detected at both seedling and germination stages. Allelic variation at candidate genes OsGSTZ1, OsGSTZ2 and OsCDPK13 was analyzed in a subset of ten contrasting lines. Ten out of thirty-four polymorphisms detected in all three genes were associated with cold tolerance in these lines. A functional marker, designed from an aminoacid-changing SNP found in gene OsGSTZ2, was tested in the 116 lines. As a result, a tight association was observed between cold tolerance and marker score. In conclusion, wide variability for cold tolerance at early developmental stages has been detected in rice advanced breeding materials that are adapted to local growing conditions. Also, a functional marker tightly associated with the trait is available for performing marker assisted selection.     

Genetic Structure of Field Populations of Begomoviruses and of Their Vector Bemisia tabaci in Pakistan

ABSTRACT The genetic structure of field populations of begomoviruses and their whitefly vector Bemisia tabaci in Pakistan was analyzed. Begomoviruses and B. tabaci populations were sampled from different crops and weeds in different locations in Punjab and Sindh provinces, in areas where cotton leaf curl disease (CLCuD) occurs or does not occur. Phylogenetic analysis based on nucleotide sequences of the intergenic region in the viral DNA-A provided evidence of two clusters of isolates: viruses isolated from species in the family Malvaceae, and viruses isolated from other dicotyledon families. Analysis of the capsid protein (CP) open reading frame grouped isolates into three geographical clusters, corresponding to isolates collected in Punjab, Sindh, or both provinces. Random amplified polymorphic DNA analyses of the B. tabaci population showed that intrapopulation diversity was high at both the local and regional scales. Sequence analysis of the mitocondrial cytochrome oxydase I (mt COI) gene showed that the B. tabaci population was structured into at least three genetic lineages corresponding to the previously described Indian, Southeast Asian, and Mediterranean-African clades. The Indian clade was present only in Punjab, the Mediterranean-African only in Sindh, and the Southeast Asian in both provinces. B. tabaci haplotypes of the Indian clade were found only in the Punjab, where CLCuD occurs. Hence, the geographical distribution of virus and vector genotypes may be correlated, because similar phylogenetic relationships were detected for the viral CP and the vector mt COI genes.     

Differential Growth Response and Minituber Production of Three Potato Cultivars Under Aeroponics and Greenhouse Bed Culture

We have evaluated and compared different methods and cultivars for the production of quality prebasic seed in potato. Two cultivation systems, aeroponics and greenhouse beds with a peat moss substrate, and three potato cultivars with different vegetative cycle, Agria, Monalisa and Zorba, were assayed. Plants in the aeroponic system showed increased growth and their vegetative cycle extended between 12 and 36 % compared to the plants cultivated in greenhouse beds. Flowering and tuberization dates, Absolute Growth Rates (AGR) during the period of 60 days after planting (DAP) and height presented a wide variation between cultivars. Zorba showed earlier flowering and tuberization, lower AGR and reached a minor height at 60 DAP. Instead the late season cultivar Agria showed later flowering and tuberization, presented higher AGR and reached an increased height at 60 DAP. The total tuber yield per plant was between 34 and 87 % higher in the aeroponic system, with a marked difference for the earlier cultivars Zorba and Monalisa. Tuber numbers increased between 60 and 80 %. Minituber production in aeroponics showed a better size distribution, with a reduction in the percentage of tubers smaller than 12 mm of between 33 and 86 %. In this soil-less culture system average tuber weight increased in Zorba and Monalisa over 60 % but was lower for Agria. Further studies are needed to optimize aeroponics system, which can be considered a high yield potato multiplication system, particularly for early or mid season potato cultivars that may produce best quality minitubers.