To pool or not to pool? Guidelines for pooling samples for use in surveillance testing of infectious diseases in aquatic animals

Samples from multiple animals may be pooled and tested to reduce costs of surveillance for infectious agents in aquatic animal populations. The primary advantage of pooling is increased population‐level coverage when prevalence is low (<10%) and the number of tests is fixed, because of increased likelihood of including target analyte from at least one infected animal in a tested pool. Important questions and a priori design considerations need to be addressed. Unfortunately, pooling recommendations in disease‐specific chapters of the 2018 OIE Aquatic Manual are incomplete and, except for amphibian chytrid fungus, are not supported by peer‐reviewed research. A systematic review identified only 12 peer‐reviewed aquatic diagnostic accuracy and surveillance studies using pooled samples. No clear patterns for pooling methods and characteristics were evident across reviewed studies, although most authors agreed there is a negative effect on detection. Therefore, our purpose was to review pooling procedures used in published aquatic infectious disease research, present evidence‐based guidelines, and provide simulated data examples for white spot syndrome virus in shrimp. A decision tree of pooling guidelines was developed for use by peer‐reviewed journals and research institutions for the design, statistical analysis and reporting of comparative accuracy studies of individual and pooled tests for surveillance purposes.     

Patterns of SSR variation in bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) seeds under ex situ genebank storage and accelerated ageing

Maintaining seed viability and germplasm integrity is a challenging task in conservation of plant genetic resources, as seeds under storage will lose viability and genetic changes will occur. Attempt was made to analyze the patterns of genetic changes in wheat germplasm under ex situ genebank storage and accelerated ageing treatments. A set of 16 naturally aged wheat accessions under ex situ genebank storage since 1994 were sampled. Four recently regenerated wheat accessions were selected, four random seed samples were chosen from each accession, and three of them were exposed to three different accelerated ageing treatments. These 32 seed samples in two germplasm sets displayed a range of germination rates from 4 to 98 %. Thirty-seven microsatellite markers representing 21 wheat chromosomes were applied to screen 12 seeds of each sample and 449 SSR alleles were scored. Large SSR variation was found in each germplasm set. There was 73.1 % of the total SSR variation present among the naturally aged samples and 78.2 % present among the accelerated ageing samples. Several analyses for genetic association consistently revealed no clear genetic separations among samples of high or low germination rates in both germplasm sets. Samples under different accelerated ageing treatments did not show much genetic differentiations from the original sample of each accession. Mantel tests revealed non-significant associations between SSR variability and sample germination rates for both germplasm sets. These findings are useful for understanding seed deterioration under different ageing conditions and suggest that genome-wide SSR variability may not provide sensitive markers for the monitoring of wheat seed viability.     

Effects of growing roots of Norway spruce (Picea abies [L.] Karst.) and European beech (Fagus sylvatica L.) on rhizosphere soil solution chemistry

The chemical conditions of the rhizosphere can be very different from that of bulk soil. Up to now, little attention has been given to the problem of spatial heterogeneity and temporal dynamics of rhizosphere soil solution and little is known about the influence of different tree species on rhizosphere chemistry. In the present study, we used micro suction cups to collect soil solution from the rhizosphere of Norway spruce (Picea abies [L.] Karst.) and European beech (Fagus sylvatica L.) seedlings in high spatial resolution and capillary electrophoresis for the determination of major cations and anions. The results indicate, that in a soil with a base saturation of about 20—25% and a pH of 6.5, growing roots of beech and spruce lower the concentrations of nutrient cations and nitrate in the rhizosphere soil solution and decrease significantly the pH. The H⁺ release leads to an enhanced mineral weathering as indicated by an increase of CEC and base saturation and to a mobilization of soluble Al, however, on a very low concentration level. In our experiment rhizosphere effects of spruce have been more pronounced than those of beech, indicating, that with respect to below ground activity young spruce trees have a better competitive power than beech.     

Single-step purification and evaluation of recombinant BP26 protein for serological diagnosis of Brucella ovis infection in rams

To investigate the value of the BP26 protein in the serological diagnosis of ovine brucellosis caused by Brucella ovis, recombinant BP26 protein was produced in Echerichia coli and purified for use in an indirect enzyme-linked immunosorbent assay (I-ELISA). The majority of the recombinant protein was recovered from the supernatant of sonicated recombinant E. coli cells in a soluble form. This facilitated the purification of the recombinant BP26 protein which was achieved by using ion-exchange chromatography. After one step of purification, the purity of the recombinant BP26 protein was analyzed by using SDS-PAGE, Coomassie blue staining, and Western blot with a monoclonal antibody (MAb) directed against the BP26 protein. The degree of purity appeared satisfactory so that it could be directly used in I-ELISA. Although the recombinant BP26-ELISA appeared less useful than I-ELISA using the B. ovis hot saline (HS) extract as antigen, the high number of sera from B. ovis infected rams found positive (90%) in the recombinant BP26-I-ELISA indicated that the BP26 protein may be an additional suitable antigen for serological diagnosis of B. ovis infection in rams.     

Connectivity of sediment transport in a semiarid environment: a synthesis for the Upper Jaguaribe Basin,Brazil

Purpose

Hydrosedimentological studies conducted in the semiarid Upper Jaguaribe Basin, Brazil, enabled the identification of the key processes controlling sediment connectivity at different spatial scales (10⁰–10⁴ km²).

Materials and methods

Water and sediment fluxes were assessed from discharge, sediment concentrations and reservoir siltation measurements. Additionally, mathematical modelling (WASA-SED model) was used to quantify water and sediment transfer within the watershed.

Results and discussion

Rainfall erosivity in the study area was moderate (4600 MJ mm ha^?1 h^?1 year^?1), whereas runoff depths (16–60 mm year^?1), and therefore the sediment transport capacity, were low. Consequently, ～60 % of the eroded sediment was deposited along the landscape, regardless of the spatial scale. The existing high-density reservoir network (contributing area of 6 km² per reservoir) also limits sediment propagation, retaining up to 47 % of the sediment at the large basin scale. The sediment delivery ratio (SDR) decreased with the spatial scale; on average, 41 % of the eroded sediment was yielded from the hillslopes, while for the whole 24,600-km² basin, the SDR was reduced to 1 % downstream of a large reservoir (1940-hm³ capacity).

Conclusions

Hydrological behaviour in the Upper Jaguaribe Basin represents a constraint on sediment propagation; low runoff depth is the main feature breaking sediment connectivity, which limits sediment transference from the hillslopes to the drainage system. Surface reservoirs are also important barriers, but their relative importance to sediment retention increases with scale, since larger contributing areas are more suitable for the construction of dams due to higher hydrological potential.     

Concentrations of minerals in the canine prostatic fluid

Current knowledge about the composition of the prostate fluid in healthy male dogs is limited and restricted to small case numbers. Furthermore, published data often vary significantly regarding sample processing and analytical methods. Therefore, we aimed to provide data on the composition of electrolytes and minerals in the canine prostatic fluid in a larger population (n = 30 dogs/samples) and to compare these results with the existing literature. Concentrations of sodium, potassium and copper analysed in our population were most consistent with those in the literature. Different to this, concentrations of total calcium, magnesium, zinc and inorganic phosphate varied. Whereas magnesium, zinc and inorganic phosphate seemed to depend on the analysis method, total calcium concentrations differed if centrifugation was performed or not. Our results clearly indicate a need for standardization of methods for analysis of seminal plasma components.     

Kennzeichnung von eigenschaften der folgeböden nordostdeutscher niedermoore

Aufgrund intensiver landwirtschaftlicher Nutzung unterliegen Niedermoore gravierenden Struktur‐ und Funktionsveränderungen. Mineralisierung führt zur Abnahme des Gehaltes an organischer Bodensubstanz bis zum vollständigen Verzehr des Torfkörpers und damit zur Entstehung von Folgeböden. Dabei erlangen die unterschiedlichen Untergrundsubstrate, Mudden und mineralische Substrate unterschiedlicher Korngrößenzusammensetzung, immer größeren Einfluß auf die Ausprägung der neu entstehenden Bodentypen. Aus Mudden hervorgehende organische Folgeböden sind teilweise als vollkommen neue Bodensubstrate anzusehen, deren Eigenschaften wenig erforscht sind. Für die aus mineralisch unterlagerten Mooren entstehenden Folgeböden ist die Entwicklung zu Anmooren und Humussanden bekannt, jedoch noch nicht näher untersucht. Es werden erste Ergebnisse von Untersuchungen zu Bodenkennwerten dieser Böden vorgestellt. Mit welcher Entstehung von Bodentypen zu rechnen ist und welche Faktoren in diesem Prozeß von Bedeutung sind, wird abgeleitet. Mögliche Nutzungsperspektiven, die sich in Nutzungs‐ und Entwicklungskonzepte für Niedermoore eingliedern, werden aufgezeigt.     

Weighing in on a method to discriminate maize haploid from hybrid seed

The doubled haploid breeding method can produce maize inbred lines faster than traditional methods, but there are challenges associated with it. Sorting haploid from hybrid seed based on visual colour markers is time consuming and can be difficult due to colour inhibitors that obscure pigmentation needed to distinguish between haploid, hybrid and outcrossed seed. In this study, weight was evaluated as a method to sort haploid from hybrid seed. A first experiment utilized two families for analysis in a preliminary study. Eleven haploid and hybrid kernels from both families were weighed for a total of 44 experimental units. A second experiment was carried out using six families, using the same format as the previous, for 132 experimental units. Hybrid seed weighed significantly more than haploid seed in both experiments. However, the interaction between line and kernel type was significant in the second experiment. In conclusion, efficacy of sorting haploid from hybrid kernels based on weight depends on the genotypes involved.