Effects of type of added salt and ionic strength on physicochemical and functional properties of casein isolates produced by electroacidification

A procedure developed for soybean protein precipitation which was based on electrodialysis was tested for the production of acid casein from reconstituted skim milk. In a previous paper, the performance of bipolar membrane electroacidification (BMEA) was evaluated under different conditions of ionic strength (micro(added) = 0, 0.25, 0.5, or 1.0 M) and added salt (CaCl(2), NaCl, or KCl) (1). The aim of this study, which is the complement of the work on evaluation of BMEA performance, was to evaluate the functionality of the protein isolates produced by BMEA and to compare the BMEA isolates to commercial isolates and an isolate produced by chemical acidification. It was not possible to show differences between the functional properties of isolates produced by BMEA, except at 1 M CaCl(2) micro(added), due to the variability of the isolates. However, the results showed that it is possible to obtain isolates similar to commercial isolates and that the addition of salt during the process does not induce variations in functional properties. From results on mineral concentrations, it appeared that the addition of monovalent cations did not influence the retention of monovalent or divalent cations in the BMEA isolates, while addition of divalent cations (CaCl(2)) influenced the retention of magnesium. According to previous results on evaluation of BMEA performances under different conditions of ionic strength and added salt, the difference observed for the BMEA isolate produced at 1.0 M CaCl(2) was confirmed.     

Tissue tropism of Helicobacter pullorum in specific pathogen-free chickens determined by culture and nucleic acid detection

Three-day-old specific pathogen-free chickens (n = 24) located in isolators were inoculated orally with Helicobacter pullorum. One group (n = 12) was infected with a H. pullorum field isolate from human origin, another one (n = 12) with the American Type Culture Collection H. pullorum reference isolate 51801 originating from chickens. Both isolates were positive for cytolethal distending toxin, investigated using a polymerase chain reaction (PCR). A third group (n = 4) was kept as a negative control. Starting on day 7 of life, birds from each group were euthanatized at different time points up to 35 days. Various organ samples were taken aseptically and processed by culture and a H. pullorum-specific PCR. In the group infected with the human isolate the nucleic acid of H. pullorum was detected in the caecal tonsils and caeca of 12 and 11 birds, respectively. Live bacteria were cultivated from the caecal tonsils and caeca of five birds 24 and 31 days postinfection. Live bacteria were also isolated from the heart of one bird, whereas PCR had to be used to detect the nucleic acid of H. pullorum in the gallbladder of four birds. No live bacteria were reisolated at any time from birds infected with the avian isolate, but bacterial nucleic acid was detected in the caeca of five birds and in the gallbladder of one. In both groups neither live H. pullorum nor its nucleic acid were detected in the liver, spleen, and duodenum. Compared to the avian H. pullorum isolate the human isolate proved to be more invasive. No obvious clinical symptoms or disease was seen in the chickens during the entire experiment. The reisolation of live bacteria at the end of the experiments indicates that H. pullorum could enter the food chain even after early infection in birds. Furthermore, PCR was demonstrated to be helpful in tracing these fastidious bacteria.     

Effect of storage time and temperature on the total protein concentration and electrophoretic fractions in equine serum

Serum protein electrophoresis (SPE) is a technique that could be considered one of the most useful diagnostic aids available to the clinician. The effect of storage time and temperature on the total proteins and electrophoretic fractions (albumin, α₁-, α₂-, β₁-, β₂-, and γ-globulins) was assessed in 24 healthy horses. All samples, collected by jugular vein puncture, were centrifuged and divided into 4 aliquots. The 1st aliquot was analyzed within 3 h from collection (time 0), the 2nd was refrigerated at +4°C for 24 h, the 3rd was refrigerated at +4°C for 48 h, and the last was frozen at −20°C for 48 h. One-way repeated-measures analysis of variance (ANOVA) showed a significant effect (P < 0.05) of the different storage conditions on the concentrations of all the parameters studied and significant variations in the percentages of albumin, α₁-globulins, α₂-globulins, and γ-globulins. Compared with time 0 the total protein concentration increased significantly after 48 h at −20°C, the albumin percentage decreased after 48 h at −20°C, the α₁-globulin percentage increased after 24 h at +4°C, the α₂-globulin percentage increased after 48 h at +4°C and at −20°C, and the γ-globulin percentage increased after 48 h at −20°C. The results should help veterinary practitioners handle and store equine serum samples appropriately. Further investigations at different storage times and temperatures could be useful.     

Molecular detection of sugarcane striate virus and sugarcane white streak virus and their prevalence in the Miami World Collection of sugarcane and related grasses

Viruses in the genus Mastrevirus (family Geminiviridae), including those infecting sugarcane, have natural geographical ranges almost exclusively restricted to Africa and the Indian Ocean islands off the African coast. Only sugarcane white streak virus (SWSV) in Barbados and sugarcane striate virus (SStrV) in Florida and Guadeloupe are known to infect a few sugarcane varieties in the Western Hemisphere. In this study, PCR assays were developed to detect these two viruses in sugarcane. Five hundred and seventy-one DNA samples from Saccharum species and interspecific hybrids from the Miami World Collection of sugarcane and related grasses were tested for the presence of SStrV and SWSV by PCR. No variety was found infected by SWSV but SStrV was detected in 19 varieties. PCR data were confirmed by sequencing amplified fragments (248 bp). These fragments shared 93%–100% nucleotide identity with SStrV sequences from the GenBank database. SStrV isolates were distributed in six phylogenetic groups, including the four strains of the virus. Most varieties infected by SStrV originated from Asia, thus confirming a previous hypothesis stating that this virus originated from this continent. Absence of SStrV in commercial sugarcane in Florida also suggested that this virus has not been spread in this location, while infected plants have been present for several decades.     

Contingency nature of Helicobacter bizzozeronii oxygen-insensitive NAD(P)H-nitroreductase (HBZC1_00960) and its role in metronidazole resistance

Genomic analysis of a metronidazole resistant H. bizzozeronii strain revealed a frame length extension of the oxygen-insensitive NAD(P)H-nitroreductase HBZC1_00960 (RdxA), associated with the disruption of the C-terminal cysteine-containing conserved region (IACLXALGK). This was the result of the extension (from C₈ to C₉) of a simple sequence cytosine repeat (SSCR) located in the 3’ of the gene. A 3'' SSCR is also present in the rdxA homolog of H. heilmannii sensu stricto, but not in H. pylori. We showed that in the majority of in vitro spontaneous H. bizzozeronii metronidazole resistant mutants, the extension of the 3′ SSCR of rdxA was the only mutation observed. In addition, we observed that H. bizzozeronii ΔrdxA mutant strain showed the same MIC value of metronidazole observed in the spontaneous mutants. These data indicate that loss of function mutations in rdxA and in particular the disruption of the conserved region IACLXALGK is associated with reduced susceptibility to metronidazole in H. bizzozeronii. Slipped-strand mispairing of the SSCR located in the 3′ of the H. bizzozeronii rdxA appears to be the main mechanism. We also observed that H. bizzozeronii acquires resistance to metronidazole at high mutation rate, and that serial passages in vitro without selection induced an increased level of susceptibility. In conclusion, contrary to what was previously described in H. pylori, the H. bizzozeronii rdxA appears to be a contingency gene which undergoes phase variation. The contingency nature of rdxA should be carefully considered when metronidazole is used in the treatment of H. heilmannii-associated gastritis.     

A predator unmasked: life cycle of Bdellovibrio bacteriovorus from a genomic perspective

Predatory bacteria remain molecularly enigmatic, despite their presence in many microbial communities. Here we report the complete genome of Bdellovibrio bacteriovorus HD100, a predatory Gram-negative bacterium that invades and consumes other Gram-negative bacteria. Its surprisingly large genome shows no evidence of recent gene transfer from its prey. A plethora of paralogous gene families coding for enzymes, such as hydrolases and transporters, are used throughout the life cycle of B. bacteriovorus for prey entry, prey killing, and the uptake of complex molecules.     

Bioindication and modelling of atmospheric deposition in forests enable exposure and effect monitoring at high spatial density across scales

Key message

Moss surveys provide spatially dense data on environmental concentrations of heavy metals and nitrogen which, together with other biomonitoring and modelling data, can be used for indicating deposition to terrestrial ecosystems and related effects across time and areas of different spatial extension.

Context

For enhancing the spatial resolution of measuring and mapping atmospheric deposition by technical devices and by modelling, moss is used complementarily as bio-monitor.

Aims

This paper investigated whether nitrogen and heavy metal concentrations derived by biomonitoring of atmospheric deposition are statistically meaningful in terms of compliance with minimum sample size across several spatial levels (objective 1), whether this is also true in terms of geostatistical criteria such as spatial auto-correlation and, by this, estimated values for unsampled locations (objective 2) and whether moss indicates atmospheric deposition in a similar way as modelled deposition, tree foliage and natural surface soil at the European and country level, and whether they indicate site-specific variance due to canopy drip (objective 3).

Methods

Data from modelling and biomonitoring atmospheric deposition were statistically analysed by means of minimum sample size calculation, by geostatistics as well as by bivariate correlation analyses and by multivariate correlation analyses using the Classification and Regression Tree approach and the Random Forests method.

Results

It was found that the compliance of measurements with the minimum sample size varies by spatial scale and element measured. For unsampled locations, estimation could be derived. Statistically significant correlations between concentrations of heavy metals and nitrogen in moss and modelled atmospheric deposition, and concentrations in leaves, needles and soil were found. Significant influence of canopy drip on nitrogen concentration in moss was proven.

Conclusion

Moss surveys should complement modelled atmospheric deposition data as well as other biomonitoring approaches and offer a great potential for various terrestrial monitoring programmes dealing with exposure and effects.

     

Short communication: detection and molecular characterization of hepatitis E virus in domestic animals of São Tomé and Príncipe

Tropical Animal Health and Production - As in most of the African continent, the status of hepatitis E virus (HEV) infection in domestic animals in São Tomé and Príncipe, an...     

EFFECTS OF FOLIAR AND SOIL CALCIUM APPLICATION ON YIELD AND QUALITY OF TABLE GRAPE CV. ‘THOMPSON SEEDLESS’

Table grape quality and firmness are key parameters for Chile's export, given distances from the main markets. As a consequence, calcium (Ca) fertilization has become a standard practice in the country, although its effects have not been fully evaluated. The objectives in this study were to investigate the effects of calcium fertilization on table grape nutrition and quality. A two-year field trial was conducted in central Chile, using grapevines (Vitis vinifera) cv. ‘Thompson seedless’, grown on an overhead trellis system. Calcium (CaCl₂) application treatments included foliar, soil, foliar plus soil, and a control without Ca fertilization. Calcium and other nutrients concentration were determined afterwards in soil, leaves, and berries. Also, yield and some fruit quality parameters were evaluated. Calcium application method did not have an effect on soil, leaf or fruit Ca content, neither on yield and berry sugar concentration, with levels being similar to the control treatment. Additionally, in treatments where CaCl₂ was applied to soil, chlorides concentration in soil, leaves, and berries were higher. Also, berries were larger, more turgent, with lower dry matter and larger cells. In the second season, in these same treatments, berry firmness at harvest was higher, but this effect disappeared after the storage simulation period. These results question the validity that Ca applications might improve table grape postharvest quality for growing conditions similar to the ones in this experiment. The temporal changes in berry quality parameters were attributed to higher Cl soil content and the associated plant osmoregulatory mechanisms.