A combined immunomagnetic separation and lateral flow method for a sensitive on-site detection of Bacillus anthracis spores – assessment in water and dairy products

Aim:  Combination of immunomagnetic separation (IMS) and lateral flow device (LFD) assays for the development of a sensitive, rapid, on-site methodology that enables concentration and detection of Bacillus anthracis spores in complex samples.
Methods and Results:  The data presents the development of an optimized, 30 min, IMS assay, with about 95% capture of B. anthracis spores from different dairy products ( n  = 38). No cross reactivity was detected with typical milk flora and some closely related Bacilli. To enable direct application of the IMS captured spores on the LFD, spores were eluted from the bead–spore complex utilizing 95% (v/v) formamide-10 mmol l⁻¹ EDTA for 30 s in a microwave oven. Detached spores were analysed on LFD enabling detection within 10 min. The combined IMS–LFD methodology (40 min) demonstrates a 60-fold improvement in sensitivity, relative to samples that were applied directly on the LFD without the IMS concentrating step.
Conclusions:  The IMS–LFD method is a powerful platform, combining rapidity, specificity and efficiency for concentrating and detecting B. anthracis from water and milk contaminated samples.
Significant and Impact of the Study:  The combination of IMS and LFD enhances the sensitivity and flexibility of B. anthracis spore detection from complex samples. This method can potentially be extended to other toxins and micro-organisms in a variety of matrices.     

Presenilin-1 Dependent Neurogenesis Regulates Hippocampal Learning and Memory

Presenilin-1 (PS1), the catalytic core of the aspartyl protease γ-secretase, regulates adult neurogenesis. However, it is not clear whether the role of neurogenesis in hippocampal learning and memory is PS1-dependent, or whether PS1 loss of function in adult hippocampal neurogenesis can cause learning and memory deficits. Here we show that downregulation of PS1 in hippocampal neural progenitor cells causes progressive deficits in pattern separation and novelty exploration. New granule neurons expressing reduced PS1 levels exhibit decreased dendritic branching and dendritic spines. Further, they exhibit reduced survival. Lastly, we show that PS1 effect on neurogenesis is mediated via β-catenin phosphorylation and notch signaling. Together, these observations suggest that impairments in adult neurogenesis induce learning and memory deficits and may play a role in the cognitive deficits observed in Alzheimer’s disease.     

Long-term automatic trap data reveal factors affecting diurnal flight patterns of the Mediterranean Fruit fly

Mediterranean Fruit Fly (medfly), Ceratitis capitata (Wiedemann), is considered to be one of the most destructive fruit insect pests throughout the world. In recent years, the temporal dynamics of the medfly have been extensively studied by season, and, to a lesser extent, on a daily basis. We exploited, for the first time, a sequential automatic medfly male trapping system, in combination with climate data, to characterize diel flight patterns of male medflies trapped. The process was carried out in four commercial citrus fields over three growing seasons. Results showed that throughout the year, medfly flight activity began 4–6 hr after sunrise and peaked up to 2 to 3 hr before sunset. Generalized additive models (GAM) of the 934 days sampled revealed that in the fall, the diurnal flight activity was unimodal, that is, it began increasing at noon, peaking in the afternoon. By contrast, in the spring and summer, the diurnal flight activity exhibited a bimodal pattern, decreasing at midday. GAMs also revealed that hour after sunrise and temperature influenced medfly captures, with the former the more dominant factor. Thus, photophase significantly impacted the medfly's diurnal flight activity in each season. This study demonstrated that automatic traps that timestamp each capture are a primary research tool in insect flight activity studies that contributes to understanding diurnal insect activity within the growing season.     

Epidemiological,clinical and laboratory features of acute Q fever in a cohort of hospitalized patients in a regional hospital,Israel, 2012-2018

IntroductionAcute Q fever is endemic in Israel, yet the clinical and laboratory picture is poorly defined.MethodsA retrospective study reviewing the medical records of acute Q fever patients, conducted in a single hospital in the Sharon district, Israel. Serum samples from suspected cases were preliminary tested by a qualitative enzyme immunoassay (EIA). Confirmatory testing at the reference laboratory used an indirect immunofluorescence assay (IFA). Positive cases were defined as fever with at least one other symptom and accepted laboratory criteria such as a single-phase II immunoglobulin G (IgG) antibody titer ≥1:200. Cases not fulfilling these criteria and in which acute Q fever was excluded, served as a control group.ResultsBetween January 2012 and May 2018, 484 patients tested positive. After confirmatory testing, 65 (13.4%) were positive for acute Q fever (with requisite clinical picture), 171 (35.3%) were definitely not infected, the remaining 248 were excluded because of past/chronic/undetermined infection. The average age was 58 years and 66% were males. Most resided in urban areas with rare animal exposure. Pneumonia was seen in 57% of cases and a combination with headache/hepatitis was highly suggestive of acute Q fever diagnosis. Syncope/presyncope, fall and arthritis were more common in acute Q fever cases. Laboratory indexes were similar to the control group, except for erythrocyte sedimentation rate (ESR) which was more common and higher in the study group.ConclusionAcute Q fever in the Sharon district could be better diagnosed by using a syndromic approach in combination with improved rapid diagnostic testing.     

Renin-aldosterone response, urinary Na/K ratio and growth in pseudohypoaldosteronism patients with mutations in epithelial sodium channel (ENaC) subunit genes

Multi-system pseudohypoaldosteronism (PHA) is a rare syndrome of aldosterone unresponsiveness characterized by symptoms of severe salt-losing caused by mutations in one of the genes that encode alpha, beta or gamma subunit of epithelial sodium channels (ENaC). We examined long-term changes in the renin-aldosterone response in patients with different mutations. Four PHA patients were followed-up for 7-22 years. Patient A with a heterozygous Gly327Cys missense mutation in alphaENaC is a mild case and patients B, C and D are severe cases. Two additional patients with renal PHA served as controls. In patient A, serum aldosterone and plasma renin activity (PRA) decreased with age, PRA reaching near normal values at age 11. In contrast, patients B-D showed a positive correlation between age and aldosterone (r>0.86 for all). In patient B with Arg508 stop mutation, aldosterone reached 166nmol/L at age 19 (>300-fold higher than normal). Urinary Na/K ratios normalized gradually with age in all patients. Growth curves of the patients were reflective of the severity of PHA and compliance with salt therapy. Functional expression studies in oocytes showed that ENaC with alphaGly327Cys mutation, as observed in patient A, showed nearly 40% activity of the wild type ENaC. In contrast, stop mutation as in patient B reduces ENaC activity to less than 5% of the normal. Our results demonstrate distinct genotype-phenotype relationships in multi-system PHA patients. The degree of ENaC function impairment affects differently the renin-aldosterone system and urinary Na/K ratios. The differences observed are age-dependent and PHA form specific.     

Empirical evidence of the mediterranean fruit fly movement between orchard types

Understanding species movement in the agroecological system is an important theme in ecology. A mark–release–capture experiment was conducted to study the dispersion behaviour of the Mediterranean fruit fly, Ceratitis capitata (Wied.; Diptera: Tephritidae; Medfly) in northern Israel. Four pairs of pear and citrus orchards were selected for the field experiments. Sterile flies dyed with different colours were released in three seasons during 2015 and 2016, based on the phenological stages of the hosts. The total number of captured marked sterile flies per trap (FT) was approx 300 in both April and August. In November, FT values decreased by about 35% to approximately 200. The wild Medfly that were also captured showed an opposite trend, from an FT of 1.8 and 6.4 in April and August, respectively, to an FT of about 330 in November. Marked sterile flies were captured in both the release and the neighbouring orchard sites. We found that the Medfly migrates from pear to citrus orchards during spring and from citrus to pear during summer, when there are no fruit-bearing trees in the orchards. However, during November, when the wild Medfly population prevails in the area, a clear pattern of migration is hard to identify, perhaps because of a possible interaction with the wild fly's population.     

Inconsistent effects of local and landscape factors on two key pests in Israeli vineyards

An ecoinformatics approach was used to test how two noxious species in grapevines with varying host preferences and movement characteristics: the European grapevine moth (Lobesia botrana), and a parasitic fungus, the grape powdery mildew (Uncinula necator), are affected by local and landscape variables. Data were collected from 202 vineyards during four seasons: 2013, 2014, 2016 and 2017 and analysed using generalized mixed models. We hypothesized that the European grapevine moth would be affected most by the landscape variables, while the grape powdery mildew would be affected most by local ones. We found that the number of sprayings during the season was an important variable explaining variation in infestation levels between vineyards for both species. At the landscape scale, we found larger variations in the relationship between the studied variables and the two pests, but both were also affected by the surrounding land use including areas of deciduous orchards. Understanding the factors that affect the occurrence of various pests in the same environment might improve farmers' decision-making.