Properties of an alkali-thermo stable xylanase from Geobacillus thermodenitrificans A333 and applicability in xylooligosaccharides generation

An extracellular thermo-alkali-stable and cellulase-free xylanase from Geobacillus thermodenitrificans A333 was purified to homogeneity by ion exchange and size exclusion chromatography. Its molecular mass was 44 kDa as estimated in native and denaturing conditions by gel filtration and SDS-PAGE analysis, respectively. The xylanase (GtXyn) exhibited maximum activity at 70 °C and pH 7.5. It was stable over broad ranges of temperature and pH retaining 88 % of activity at 60 °C and up to 97 % in the pH range 7.5–10.0 after 24 h. Moreover, the enzyme was active up to 3.0 M sodium chloride concentration, exhibiting at that value 70 % residual activity after 1 h. The presence of other metal ions did not affect the activity with the sole exceptions of K+ that showed a stimulating effect, and Fe2+, Co2+ and Hg2+, which inhibited the enzyme. The xylanase was activated by non-ionic surfactants and was stable in organic solvents remaining fully active over 24 h of incubation in 40 % ethanol at 25 °C. Furthermore, the enzyme was resistant to most of the neutral and alkaline proteases tested. The enzyme was active only on xylan, showing no marked preference towards xylans from different origins. The hydrolysis of beechwood xylan and agriculture-based biomass materials yielded xylooligosaccharides with a polymerization degree ranging from 2 to 6 units and xylobiose and xylotriose as main products. These properties indicate G. thermodenitrificans A333 xylanase as a promising candidate for several biotechnological applications, such as xylooligosaccharides preparation

A comparison of polarized and non-polarized human endometrial monolayer culture systems on murine embryo development

BACKGROUND: Co-culture of embryos with various somatic cells has been suggested as a promising approach to improve embryo development. Despite numerous reports regarding the beneficial effects of epithelial cells from the female genital tract on embryo development in a co-culture system, little is known about the effect of these cells when being cultured under a polarized condition on embryo growth. Our study evaluated the effects of in vitro polarized cells on pre-embryo development. METHODS: Human endometrial tissue was obtained from uterine specimens excised at total hysterectomy performed for benign indications. Epithelial cells were promptly isolated and cultured either on extra-cellular matrix gel (ECM-Gel) coated millipore filter inserts (polarized) or plastic surfaces (non-polarized). The epithelial nature of the cells cultured on plastic was confirmed through immunohistochemistry, and polarization of cells cultured on ECM-Gel was evaluated by transmission electron microscopy (TEM). One or two-cell stage embryos of a superovulated NMRI mouse were then flushed and placed in culture with either polarized or non-polarized cells and medium alone. Development rates were determined for all embryos daily and statistically compared. At the end of the cultivation period, trophectoderm (TE) and inner cell mass (ICM) of expanded blastocysts from each group were examined microscopically. RESULTS: Endometrial epithelial cells cultured on ECM-Gel had a highly polarized columnar shape as opposed to the flattened shape of the cells cultured on a plastic surface. The two-cell embryos cultured on a polarized monolayer had a higher developmental rate than those from the non-polarized cells. There was no statistically significant difference; still, the blastocysts from the polarized monolayer, in comparison with the non-polarized group, had a significantly higher mean cell number. The development of one-cell embryos in the polarized and non-polarized groups showed no statistically significant difference. CONCLUSION: Polarized cells could improve in vitro embryo development from the two-cell stage more in terms of quality (increasing blastocyst cellularity) than in terms of developmental rate

Routine health management information system data in Ethiopia: consistency, trends, and challenges.

Background: Ethiopia is investing in the routine Health Management Information System. Improved routine data are needed for decision-making in the health sector. Objective: To analyse the quality of the routine Health Management Information System data and triangulate with other sources, such as the Demographic and Health Surveys. Methods: We analysed national Health Management Information System data on 19 indicators of maternal health, neonatal survival, immunization, child nutrition, malaria, and tuberculosis over the 2012-2018 time period. The analyses were conducted by 38 analysts from the Ministry of Health, Ethiopia, and two government agencies who participated in the Operational Research and Coaching for Analysts (ORCA) project between June 2018 and June 2020. Using a World Health Organization Data Quality Review toolkit, we assessed indicator definitions, completeness, internal consistency over time and between related indicators, and external consistency compared with other data sources. Results: Several services reported coverage of above 100%. For many indicators, denominators were based on poor-quality population data estimates. Data on individual vaccinations had relatively good internal consistency. In contrast, there was low external consistency for data on fully vaccinated children, with the routine Health Management Information System showing 89% coverage but the Demographic and Health Survey estimate at 39%. Maternal health indicators displayed increasing coverage over time. Indicators on child nutrition, malaria, and tuberculosis were less consistent. Data on neonatal mortality were incomplete and operationalised as mortality on day 0-6. Our comparisons with survey and population projections indicated that one in eight early neonatal deaths were reported in the routine Health Management Information System. Data quality varied between regions. Conclusions: The quality of routine data gathered in the health system needs further attention. We suggest regular triangulation with data from other sources. We recommend addressing the denominator issues, reducing the complexity of indicators, and aligning indicators to international definitions

Traditional enset (Ensete ventricosum) fermentation in the Gamo highlands of Ethiopia: physiochemical and microbial dynamics

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The use of Saccharomyces cerevisiae to suppress the effects of aflatoxicosis in broiler chicks.

Three hundred and sixty day-old commercial broiler chicks were assigned to a 3 x 2 factorial arrangement of treatments to examine the effects of three levels of Saccharomyces cerevisiae 0, .05, and .1% of feed, and two levels of aflatoxin, 0 and 5 ppm, singly and in combination on aflatoxicosis. Each treatment group was replicated three times. The S. cerevisiae and aflatoxin were incorporated in a standard commercial broiler ration and fed to chicks for 4 wk. Data were collected weekly on body weights, and at the end of the experiment on the relative weights of the liver, proventriculus, pancreas, and heart. Serum concentrations of the albumin, total protein, cholesterol, uric acid, triglycerides, and enzyme activities of alanine transaminase, aspartate aminotransferase, lactase dehydrogenase, and creatine phosphokinase were also evaluated. Results showed that chicks receiving aflatoxin-contaminated feed had suppressed body weight (456 g), which significantly improved (516 g) with the inclusion of .1% S. cerevisiae. The relative weights of liver (3.58%), heart (.916%), and proventriculus (.770%), which increased significantly with the addition of 5 ppm of aflatoxin, were restored to 3.00, .783, and .680%, respectively, with the dietary inclusion of .1% S. cerevisiae. The serum concentrations of albumin and total protein (.66 and 1.62 g/100 mL), which were significantly decreased by aflatoxin, were elevated to .88 and 2.24 g/100 mL, respectively, with the inclusion of .1% S. cerevisiae.(ABSTRACT TRUNCATED AT 250 WORDS

Biochemical and immunological properties of related small ruminant trophoblast interferons

The objectives of this study were 1) to evaluate the biochemical and immunological properties of caprine interferon tau (cIFNγ), 2) to determine if intrauterine injection of recombinant ovine interferon tau (roIFNγ) extends CL life span in goats, and 3) to evaluate potential side effects of intramuscular (im) administration roIFNγ. Caprine IFNγ was purified, and its effects on lymphocyte proliferation were evaluated. Incorporation of 3H- thymidine into newly synthesized DNA was suppressed (P\u3c0.05) by clFNγ. Spanish goats were also fitted with bilateral uterine catheters at Day 7 or 8 postestrus. The goats received twice-daily intrauterine injections of 100 μg roIFNγ (n = 4) or caprine serum proteins (n = 4) from Days 14 to 18 postestrus. Intrauterine injection of roIFNγ extended CL life span compared with that of control goats (26.4 ± 1.7 vs 17.8 ± 1.9 d, respectively; P\u3c0.01). Potential side effects of intramuscular injections of roIFNγ were also evaluated. Goats received 0, 1, 2 or 4 mg of roIFNγ on Days 10, 13, 16 or 19 of the estrous cycle. Treatment of goats with roIFN resulted in hyperthermia (P\u3c0.01), with rectal temperatures of 40.5δ C recorded after 4 h and returning to normal (38.5δ C) after 24 h. Concomitant with the increase in rectal temperatures was a decrease (P\u3c0.01) in plasma progesterone concentrations. Therefore, the tau interferons of goats and sheep have similar biological properties and roIFNγ has side effects associated with other classes of interferons

Fermentation of enset (Ensete ventricosum) in the Gamo highlands of Ethiopia: Physicochemical and microbial community dynamics

Enset (Ensete ventricosum) provides staple food for 15 million people in Ethiopia after fermentation into kocho. The fermentation process has hardly been investigated and is prone to optimization. The aim of this study was to investigate the physicochemical and microbial dynamics of fermentation practices in the Gamo highlands. These practices show local variation, but two steps were omnipresent: scraping of the pseudostem and fermenting it in a pit or a bamboo basket. Enset plants were fragmented and fermented for two months in order to investigate the physicochemical (temperature, moisture content, pH and titratable acidity) and microbial dynamics (total viable aerobic counts, counts of Enterobacteriaceae, lactic acid bacteria, yeasts and moulds and Clostridium spores counts, and Illumina Miseq sequencing). Samples were taken on days 1, 7, 15, 17, 31 and 60. The pH decreased, whereas the titratable acidity increased during fermentation. Of all counts those of lactic acid bacteria and Clostridium spores increased during fermentation. Leuconostoc mesenteroides initiated the fermentation. Later on, Prevotella paludivivens, Lactobacillus sp. and Bifidobacterium minimum dominated. These three species are potential candidates for the development of a starter culture.status: publishe