Inoculation Level Research Articles

Production of edible flowers: irrigation and biotechnology

Garden pansy is a versatile gardening plant – it produces beautiful colorful edible-flowers with high value in gourmet cuisine. The use of irrigation and biotechnology in garden pansy cultivation can provide gains in flower productivity and nutritional value. The goal of this study was to evaluate the growth and edible flower production in garden pansy plants, submitted to different levels of irrigation and mycorrhizal inoculation. The experiment was conducted in randomized blocks in the 2 x 5 factorial design, with the presence and absence of mycorrhizal inoculation in combination with 5 levels of irrigation with 6 replicates, in a greenhouse. There was no significant interaction between the factors mycorrhizal inoculation and irrigation levels by the F test. Under the tested conditions, the mycorrhizal inoculation was unable to provide significant changes in the growth, development and flowering of garden pansy plants. It was concluded that no symbiotic efficiency was pointed out between the mycorrhizal fungus used and garden pansy plants. The best growth and yield results for cultivating and producing edible flowers of garden pansy were obtained at the 100% replenishment level of water evaporation.

Efficacy of Antimicrobial Spray Treatments in Reducing Salmonella enterica Populations on Chilled Pork

Studies reporting on alternative antimicrobial interventions for pathogen control on chilled pork carcasses and cuts are limited. In this study, the antimicrobial effects of various spray treatments against Salmonella enterica inoculated on skin-on pork samples were evaluated. Chilled pork jowls were portioned (10 by 5 by 1 cm) and inoculated, on the skin side, with a mixture of six S. enterica serotype strains to target levels of 6 to 7 log CFU/cm2 (high inoculation level) or 3 to 4 log CFU/cm2 (low inoculation level). Samples were then left nontreated (control) or were treated (10 s) using a laboratory-scale spray cabinet with water, formic acid (1.5%), a proprietary blend of sulfuric acid and sodium sulfate (SSS, pH 1.2), peroxyacetic acid (PAA, 400 ppm), or PAA (400 ppm) that was pH-adjusted (acidified) with acetic acid (1.5%), formic acid (1.5%), or SSS (pH 1.2). Samples (n = 6) were analyzed for Salmonella populations after treatment application (0 h) and after 24 h of refrigerated (4°C) storage. Irrespective of inoculation level, all spray treatments effectively reduced (P < 0.05) Salmonella levels immediately following their application. Overall, pathogen reductions for the chemical treatments, compared to the respective high and low inoculation level nontreated controls, ranged from 1.2 to 1.9 log CFU/cm2 (high inoculation level) and 1.0 to 1.7 log CFU/cm2 (low inoculation level). Acidification of PAA with acetic acid, formic acid, or SSS did not (P ≥ 0.05) enhance the initial bactericidal effects of the nonacidified PAA treatment. Salmonella populations recovered from all treated samples following 24 h of storage were, in general, similar (P ≥ 0.05) or up to 0.6 log CFU/cm2 lower (P < 0.05) than those recovered from samples analyzed immediately after treatment application. The results of the study may be used by processing establishments to help identify effective decontamination interventions for reducing Salmonella contamination on pork.

Impact of inorganic iron and haem on the human gut microbiota; An in vitro batch-culture approach.

Although iron is an essential nutrient for humans, as well as for almost all other organisms, it is poorly absorbed (~15%) from the diet such that most passes through the upper gut into the large intestine. The colonic microbiota is thus exposed to, and potentially influenced by, such residual iron which could have an impact on human health. The aim of the research described here is to determine how the major forms of dietary iron (inorganic iron and haem) influence metabolic activity and composition of the human gut microbiota by utilizing an in vitro parallel, pH-controlled anaerobic batch culture approach. Controlled iron provision was enabled by the design of a 'modified' low-iron gut-model medium whereby background iron content was reduced from 28 to 5 μM. Thus, the impact of both low and high levels of inorganic and haem iron (18-180 μM and 7.7-77 μM, respectively) could be explored. Gut-microbiota composition was determined using next generation sequencing (NGS) based community profiling (16S rRNA gene sequencing) and flow-fluorescent in situ hybridization (FISH). Metabolic-end products (organic acids) were quantified using gas chromatography (GC) and iron incorporation was estimated by inductively coupled plasma optical emission spectroscopy (ICP-OES). Results showed that differences in iron regime induced significant changes in microbiota composition when low (0.1% w/v) fecal inoculation levels were employed. An increase in haem levels from 7.7 to 77 μM (standard levels employed in gut culture studies) resulted in reduced microbial diversity, a significant increase in Enterobacteriaceae and lower short chain fatty acid (SCFA) production. These effects were countered when 18 μM inorganic iron was also included into the growth medium. The results therefore suggest that high-dietary haem may have a detrimental effect on health since the resulting changes in microbiota composition and SCFA production are indicators of an unhealthy gut. The results also demonstrate that employing a low inoculum together with a low-iron gut-model medium facilitated in vitro investigation of the relationship between iron and the gut microbiota.

Influence of FeS2 and FeSi75 in Double-Chamber Sample Cup on Thermal Analysis Curve and Graphite Morphology of Molten Iron

In the production of vermicular graphite cast iron, the allowable range of residual magnesium content in molten iron after treatment is very narrow, amounting to only 0.008%. Therefore, thermal analysis technology was used to quickly evaluate the vermiculation and inoculation level of molten iron at the furnace itself, thus allowing the molten iron to be adjusted in time. The additives in the sample cups play a crucial role in obtaining cooling curves with remarkable characteristics. In this study, either FeS2 or FeSi75 additives were added to one chamber of a double-chamber sample cup made of resin sand, in which the cavities of the double chambers were spherical with diameters of 30 mm. The thermal analysis curves of molten iron in the double-chamber sample cup were acquired using a double channel temperature recorder, and the solidified spherical samples were analyzed quantitatively. The influence of FeS2 or FeSi75 additives on both the cooling curves of molten iron and the graphite morphology were investigated. The experiment’s results indicated that when 0.05% FeS2 is added to one chamber of the sample cup, the cooling curve changes to the solidification pattern of gray cast iron. The continuous increase in the FeS2 additive has little influence on the shape of cooling curves, and the graphite changes form from vermicular to flaked. When the amount of FeS2 is increased from 0.05% to 0.10%, the resulting graphite changes from D-type and E-type to A-type and B-type. When the amount of FeS2 reaches 0.20%, the morphology of graphite is short and thick. With the increase in the amount of FeSi75 additive, the amount of spherical graphite in the sample cup increases gradually, and the vermicularity decreases gradually from 89% to 46%. With the increase in FeSi75 additive from 0 to 0.45%, we observed that the average diameter of graphite decreases from 23 μm to 19 μm and then increases to 22 μm. The eutectic recalescence temperature shows a decreasing trend, and the cooling curve gradually changes from a hypoeutectic to a eutectic pattern. The addition of 0.05% FeS2 or 0.45% FeSi75 to one chamber is more appropriate for a double-chamber sample cup with two spherical cavities with diameters of 30 mm. This lays a foundation for the optimization of additives when using the double-chamber sample cup for thermal analysis of vermicular graphite cast iron.

Rapid Production Biofloc by Inoculating Chlorella pyrenoidosa in a Separate Way

Microalgae play an important role in the formation of biofloc. To demonstrate the feasibility of Chlorella pyrenoidosa in biofloc formation, an experiment was performed with a simple random design consisting of five inoculation levels (in triplicate) of C. pyrenoidosa (0, 1 × 108, 1 × 109, 5 × 109, and 1 × 1010 cells·L−1) in the biofloc system. All treatments kept a C:N ratio of approximately 15:1. This study observed the effects of different initial concentrations of C. pyrenoidosa on biofloc formation, water quality and bacterial community in biofloc systems. The results indicated that C. pyrenoidosa had the ability to enhance biofloc development, especially when the C. pyrenoidosa initial concentration reached 5~10 × 109 cells·L−1. Too high or too low a concentration of C. pyrenoidosa will adversely affect the formation of biofloc. The effect of C. pyrenoidosa addition on water quality (TAN, NO2−-N, and NO3−-N) was not significant in the final stage. The inoculation of C. pyrenoidosa decreased the species richness and diversity of the bacterial community but increased the domination of Proteobacteria and Bacteroidota in the biofloc system, especially the order of Rhizobiales. The addition of C. pyrenoidosa could maintain water quality by increasing the proportion of several denitrifying bacteria, including Flavobacterium, Chryseobacterium, Pseudomonas, Brevundimonas, Xanthobacter, etc. These above dominant denitrifying bacteria in the biofloc system could play a major role in reducing the concentration of NO2−-N and NO3−-N. So, we recommended the reasonable concentration is 5~10 × 109 cells·L−1 if C. pyrenoidosa is used to rapidly produce biofloc.

Development and Verification of a Poultry Management Tool to Quantify Salmonella from Live to Final Product Utilizing RT-PCR.

The United States Department of Agriculture Food Safety and Inspection Service (USDA FSIS) does not maintain a zero-tolerance policy for Salmonella in poultry and poultry products, despite being a known food safety hazard throughout the poultry industry. In 2016, USDA FSIS established performance standards for a 52-week moving window with the maximum acceptable percent positive for comminuted turkey (325 g sample) at 13.5% (7 of 52 samples). Based upon FSIS verification sampling results from one 52-week moving window, the Salmonella prevalence for each poultry establishment in category 1 (below limit), 2 (meeting limit), or 3 (exceeding limit) are published for public viewing. Moreover, many poultry producers continue to have post-intervention samples test positive. Therefore, the use of quantification would be more valuable to determine the efficacy of process control interventions, corrective actions, and final product Log CFU/g of Salmonella to make rapid, within shift, food safety decisions. Therefore, the objectives of these studies are to develop, verify, and validate a rapid and reliable quantification method utilizing RT-PCR to enumerate Salmonella in the poultry industry from flock to final product and to utilize the method in an application study. BAX® System SalQuant® is an application of the BAX® System Real-Time PCR Assay for Salmonella to enumerate low levels of Salmonella with shortened enrichment times. Curve development encompassed inoculating poultry matrix samples at four levels with an ATCC strain of Salmonella, with three biological replicates per inoculation level, and five technical replicates being run on the BAX® System for various timepoints, gathering the data, and creating a linear-fit equation. A linear-fit equation was provided for each timepoint. The ideal timepoint, based on the statistical parameters surrounding the equation (R2 &gt; 0.80, Log RMSE &lt; 0.60, and enumerable range 0.00 to 4.00 Log CFU/mL (g)) that most accurately estimate Salmonella compared to most probable number (MPN), was chosen to be utilized for further studies.

Visual detection of Cronobacter sakazakii on a microfluidic chip fabricated by a 3D molding method.

Cronobacter sakazakii (C. sakazakii) is a pathogenic bacterium associated with life-threatening neonatal infections that have been linked to contaminated powdered infant formula (PIF). Most C. sakazakii testing is still limited in microbiology laboratories due to the need for sophisticated equipment and professional technicians. Microfluidic chips combined with isothermal amplification analysis are shown to be one of the most attractive microbiological on-site detection platforms. In this study, PDMS microfluidic chips were fabricated by a simple 3D molding method and sealed with "PDMS glue". The chip consisted of an inlet, a microchannel, six reaction wells, and six vent holes. And based on the 16S rRNA and ITS genes of C. sakazakii, we have successfully proposed a multiplex competitive annealing mediated isothermal amplification (mCAMP) assay on the microfluidic chip for the visual detection of C. sakazakii in PIF samples. The primers were fixed in the reaction wells of the chip before detection, which can be preserved for 60 days at 4 °C. The results showed that the established mCAMP assay had high specificity, and the limit of detection was 2.2 × 103 CFU g-1. With enrichment culture, even if the initial inoculation level is 1 CFU g-1, the mCAMP assay can still detect the presence of C. sakazakii in spiked PIF samples. The test results are visible to the naked eye, which is suitable for rapid analysis in resource-limited settings.

Trypanosoma brucei rhodesiense Inhibitor of Cysteine Peptidase (ICP) Is Required for Virulence in Mice and to Attenuate the Inflammatory Response.

The protozoan Trypanosoma brucei rhodesiense causes Human African Trypanosomiasis, also known as sleeping sickness, and penetrates the central nervous system, leading to meningoencephalitis. The Cathepsin L-like cysteine peptidase of T. b. rhodesiense has been implicated in parasite penetration of the blood-brain barrier and its activity is modulated by the chagasin-family endogenous inhibitor of cysteine peptidases (ICP). To investigate the role of ICP in T. b. rhodesiense bloodstream form, ICP-null (Δicp) mutants were generated, and lines re-expressing ICP (Δicp:ICP). Lysates of Δicp displayed increased E-64-sensitive cysteine peptidase activity and the mutant parasites traversed human brain microvascular endothelial cell (HBMEC) monolayers in vitro more efficiently. Δicp induced E-selectin in HBMECs, leading to the adherence of higher numbers of human neutrophils. In C57BL/6 mice, no Δicp parasites could be detected in the blood after 6 days, while mice infected with wild-type (WT) or Δicp:ICP displayed high parasitemia, peaking at day 12. In mice infected with Δicp, there was increased recruitment of monocytes to the site of inoculation and higher levels of IFN-γ in the spleen. At day 14, mice infected with Δicp exhibited higher preservation of the CD4+, CD8+, and CD19+ populations in the spleen, accompanied by sustained high IFN-γ, while NK1.1+ populations receded nearly to the levels of uninfected controls. We propose that ICP helps to downregulate inflammatory responses that contribute to the control of infection.

Effect of Different Levels of Arbuscular Mycorrhiza and Phosphorus on Productivity and Profitability of Barley (Hordeum vulgare L.)

A field experiment was carried out at Soil Conservation and Water Management Farm of Chandra Shekhar Azad University of Agriculture and Technology, Kanpur during rabi season, in 2019-20 to study the effect of different levels of Arbuscular Mycorrhiza (AM) and phosphorus on yield attributes, yield and economics of barley (Hordeum vulgare L.). The experiment was laid out in Factorial Randomized Block Design (FRBD) with 3 replications. The treatments were consisted of 4 levels of AM fungi inoculation i.e., control, AM @ 8 kg ha-1, AM @ 12 kg ha-1 and AM @ 16 kg ha-1 along with 4 phosphorus fertility level i.e., control, phosphorus @ 20 kg ha-1, phosphorus @ 30 kg ha-1 and phosphorus @ 40 kg ha-1 were tested in the experiment. The results indicated that the AM fungi inoculation level @ 16 kg ha-1 showed the highest yield attributes, yield and economics viz. number of productive tillers (9.03 m-2), number of grain ear-1 (9.16), test weight (45.36g), grain yield (28.13 q ha-1), gross return (INR 45774.20 ha-1), net return (INR 18143.04 ha-1 ) and B:C ratio (1.66) followed by the AM fungi inoculation level @ 12 kg ha-1 while it was observed minimum under control. However, the phosphorus fertility level @ 40 kg ha-1 gave better yield attributes, yield and economics viz., number of productive tillers (9.16 m-1), number of grain ear-1 (59.36), test weight (45.09g), grain yield (28.79), gross return (INR 47377.60 ha-1), net return (INR 18229.50 ha-1) and B:C ratio (1.63) followed by the phosphorus fertility level @ 30 kg ha-1 while observed minimum under control. On the basis of observed results, farmers are advised to raise barley with AM fungi inoculation level @ 16 kg ha-1 along with application of phosphorus @ 40 kg ha-1 for best growth, yield, economics and soil health.

Rapid and Visual Determination of Cronobacter sakazakii in Powdered Infant Formula Using Competitive Annealing Mediated Isothermal Amplification (CAMP)

Cronobacter sakazakii (C. sakazakii) is a foodborne pathogen that causes serious infections, particularly in infants. Neonatal infections with this pathogen have an 80% mortality rate. One mechanism that C. sakazakii infects infants is through contaminated powdered infant formula (PIF). Here, based on the 16S rRNA gene, competitive annealing-mediated isothermal amplification (CAMP) is reported for the determination of C. sakazakii to reduce the risk of infection. The established CAMP method was able to accurately identify all target bacterial strains and did not yield false positive results with non-target bacterial strains. Furthermore, accelerating primers were designed and included in the CAMP reaction to enhance the amplification efficiency. The sensitivity of the assay for C. sakazakii in spiked PIF was 1.9 × 103 CFU/g. The CAMP assay with enrichment accurately detected the presence of C. sakazakii in PIF even at an initial inoculation level of 1 CFU/g. Additionally, to make the procedure simpler, hydroxynaphthol blue was employed as a visual indicator. For positive results, the reaction solution turns blue instead of violet, while the negative samples remain violet, which is suitable for rapid analysis in resource-limited settings.

Sugarcane streak mosaic virus (ScSMV) Resistance Evaluation of Sugarcane Varieties

Sugarcane streak mosaic virus (ScSMV) is the most important viral disease of sugarcane in Indonesia with distribution in almost all commercial sugarcane plantations. The disease causes significant yield losses of both cane tonnage and sugar yield. The use of resistant varieties is the best approach for controlling viral diseases. This study aims to investigate resistance response of several introduced varieties against ScSMV in a glasshouse condition and the impact of the viral infection on chlorophyll and proline content in sugarcane leaves. Sugarcane plants were inoculated using ScSMV inoculum one month after planting using an abrasive pad rubbing method. Disease incidence and severity was observed at week 4-12 after inoculation and variety resistance levels were classified based on disease incidence. Confirmation of the virus was done by RT-PCR. Spectrophotometer was used to measure chlorophyll content at dual wavelengths of 645 and 663 nm, and proline content at wavelengths of 520 nm. The results showed that most of the tested varieties were susceptible to ScSMV. There are six highly resistant varieties, namely SRA 1, SRA 2, N 10-4, N 10-7, N10-9, and N 10-13, but these varieties still require to be tested on a field scale. ScSMV infection generally decrease chlorophyll and proline content. However, the physiological effect of ScSMV infection on chlorophyll and proline content needs further investigation.

Intercropping and Rhizobium Inoculation Affected Microclimate and Performance of Common Bean (Phaseolus vulgaris L.) Varieties.

A field experiment was carried out at Hawassa, during the 2020 cropping season with the objective to evaluate the impact of maize-common bean intercropping and Rhizobium inoculation on microclimate, growth, and yield of common bean varieties. Treatments consisting of two common bean varieties, two levels of inoculation and three spatial arrangements of common bean with another sole maize were laid out in a factorial arrangement in a randomized complete block design (RCBD) with three replications. The results revealed that the main effect of spatial arrangements highly significantly (P < 0.001) affected soil and leaf temperature. Soil moisture content was improved under intercropped plots compared with sole cropping. The intensity of light and qualities, such as red, far-red, and photosynthetically active radiations (μmol m-2 s-1) and ultraviolet rays (UV)-A, UV-B (W m-2), were reduced under intercropping as compared to the sole. Interaction effects of variety, spatial arrangements, and inoculation significantly (P < 0.01) affected plant height and leaf area index. Inoculated sole Nassir outperformed for plant height and leaf area index. Inoculated sole Hawassa Dume variety performed best for nodule number plant-1, nodule dry weight plant-1, pods number plant-1, 100 seed weight, grain yield, and above-ground biomass yield. The highest grain yield (2.8 t ha-1) was recorded from inoculated sole Hawassa Dume. However, considering the equivalent ratio (LER), intercropping with one maize row to two haricot bean rows spatial arrangements was productive by 62% more than sole cropping (total land equivalent ratio of 1.62%).

Survival Behavior of Escherichia coli O157:H7 during over 60-Day Aging of Camembert Cheeses Manufactured from Unpasteurized Raw Milk

Although many developed countries (USA, Canada, and several EU countries) allow raw milk cheese to be aged more than 60 days, these countries have strict standards for the aging conditions, such as temperature, of raw milk cheese. Spiking experiments were conducted with Camembert cheese made from raw milk, to assess the microbiological safety of raw milk cheese aged for more than 60 days. We spiked Escherichia coli O157:H7 into raw milk with different inoculation levels (high, medium, and low). Camembert cheese was prepared from the inoculated raw milk, then aged in an incubator for up to 9 weeks (63 days). There were no significant differences in pH and water activity (aW) between uninoculated cheese and cheese samples inoculated with E. coli O157:H7 (p&lt;0.05). The pH and aW of the Camembert cheese decreased throughout the storage period. In conclusion, E. coli O157:H7 did not affect the pH and aW of the cheese samples. Cell counts were conducted every week using the agar-plating method. Inoculated cells were completely eliminated, especially in Camembert cheese, after 60 days, and the reduction rate of cells was much faster in Camembert cheese.

Growth Behavior of Listeria monocytogenes during over 60-Day Ripening of Camembert Cheeses Produced by Unpasteurized Raw Milk

Although the USA, Canada, and several EU countries allow raw milk cheese to be aged more than 60 days, these countries have strict standards for its aging conditions, such as temperature. Many developed countries have employed standards of identity which effectively prevent the manufacture and sale, of cheese made from unpasteurized milk (i.e., raw milk) in interstate commerce, unless such cheese has been aged for a minimum of 60 days. The microbiological safety of raw milk Camembert cheese, aged for more than 60 days, was evaluated using spiking experiments. We spiked Listeria monocytogenes into raw milk with different inoculation levels (high, medium and low). Camembert cheese was prepared from the inoculated raw milk, then aged in an incubator for up to 9 weeks (63 days). The number of cells was determined every week using the agar-plating method. Inoculated cells were completely eliminated, especially in Camembert cheese, after 60 days, and the reduction rate of cells was much faster in Camembert cheese. There were no significant differences in pH and water activity (aW) between the uninoculated cheese and the cheese samples in which Listeria monocytogenes was inoculated (p&lt;0.05). The pH and aW of the Camembert cheese decreased throughout the storage period. In conclusion, the pathogenic bacteria used in this study did not affect the pH and aW of the Camembert cheese samples.

Use of X-ray irradiation for inactivation of Aspergillus in cannabis flower.

California cannabis regulations require testing for four pathogenic species of Aspergillus-A. niger, A. flavus, A. fumigatus and A. terreus in cannabis flower and cannabis inhalable products. These four pathogenic species of Aspergillus are important human pathogens and their presence in cannabis flower and cannabis products may pose a threat to human health. In this study, we examined the potential of X-ray irradiation for inactivation of cannabis flower contaminated with any of the four pathogenic species of Aspergillus. We determined that X-ray irradiation at a dose of 2.5 kGy is capable of rendering Aspergillus cells non-viable at low (102 spores/g dried flower), medium (103 spores/g dried flower) and high (104 spores/g dried flower) levels of inoculation. We also showed that X-ray treatment of cannabis flower did not significantly alter the cannabinoid or the terpene profiles of the flower samples. Therefore, X-ray irradiation may be a feasible method for Aspergillus decontamination of cannabis flower. More work is required to determine the consumer safety of irradiated cannabis flower and cannabis products.

Growth and nutrient uptake of haloxylon and atriplex as affected by mycorrhizal symbiosis under combined drought and salinity stresses

This study was conducted to determine the effect of arbuscular mycorrhizal fungi (AMF) symbiosis on plant growth and nutrient uptake under combined drought and salinity stresses. A pot experiment was carried out with a factorial arrangement of treatments in a completely randomized design with three replications. Experimental treatments included two plant types ( Atriplex canescens and Haloxylon ammodendron) with three levels of inoculation of fungal species ( Funneliformis geosporus, Funneliformis mosseae, and control), two levels of soil salinity stress (7 and 14 dS m–1), and two levels of drought stress (50% and 80% of management allowable depletion). Vegetative parameters, as well as root N, P, and K concentrations and uptakes, mycorrhizal growth response, mycorrhizal nitrogen response, mycorrhizal phosphorus response, mycorrhizal potassium response, and root colonization were measured. The results showed that the application of AMF increased the plant growth variables such as stem diameter, root length, shoot dry weights, and shoot to root ratio as well as nitrogen and phosphorus uptakes. The application of both AMF types was effective as compared to the control. However, F. mosseae indicated better performance especially, in terms of the effect on plant growth variables. Also, F. mosseae was more effective to relieve stress (i.e., salinity and drought) than F. geosporus. There was a significant difference between plant types and H. ammodendron had better efficiency than A. canescens under drought and salinity stresses. Based on the results, planting of H. ammodendron inoculated with F. mosseae might be recommended for soil conservation in the arid environments.

Efffects of Rhizobium Inoculation, Sulphur and Zinc Nutrients on Yield and Yield Componets of Chickpea (Cicer aritienum L.) at Gewane, Northeastern Ethiopia

&#x0D; The goal of this study was to see how a combination of Rhizobium inoculant, S, and Zn nutrients affected chickpea nodulation and yield (Cicer arientinum L.) at Gewane, northeastern Ethiopia. The experiment included 12 treatments arranged in factorial combination of levels of inoculation (Rhizobium inoculated, un-inoculated), three rates of S (0, 15, 30 kg Sulphur ha-1) and rates of Zn (0, 1.5 kg Zinc ha-1). The treatment becomes laid out in randomized complete block design with 3 replications. Analysis of variance showed that yield and yield associated parameters (main branches, number of pod, quantity of seed, hundred seed weight, grain and straw yield), crop phenology (days to 50% flowering and days to physiological maturity), nodulation scores (nodule amount, nodule volume, nodule dry weight, effective nodule and nodulation score), N and P uptake at both locations had been significantly affected by the treatments. The best (1775.5 kg ha-1) suggest rate of seed yield over locations changed into acquired from blended costs of rhizobium and 30 kg S ha-1 which led to 28 % (389 kg ha-1) growth over the control. Consequently, rhizobium inoculation with rate of 30 kg S ha-1is recommended for chickpea production on the experimental site.&#x0D;

Impact of Moisture Conservation Practices, Seed Inoculation and Zinc Level on Growth and Yield of Chickpea (Cicer arietinum L.)

A field experiment was conducted during the Rabi season of 2020-21 and 2021-22. The experiment was laid out in split-split plot design with 27 treatment combination comprising of three moisture conservation practices viz. flat bed with 2.5 t/ha crop residue, narrow bed and furrow with 2.5 t ha-1 crop residue and broad bed and furrow with 2.5 t ha-1 crop residue in main plot and three seed inoculation viz. control, rhizobium and PSB in sub plot with three zinc level viz. control, 2.5 kg Zn ha-1 and 5.0 kg Zn ha-1 in sub-sub plot. Result shows that the broad bed and furrow with 2.5 t ha-1 crop residue produced significantly all growth parameters and yields attributes as compare to flat bed with 2.5 t ha-1 crop residue and narrow bed and furrow with 2.5 t ha-1 crop residue, respectively. The all growth parameters and yields attributes were significantly recorded higher under seed inoculation with rhizobium over PSB and control. The application of different dose of zinc produced marked significant variation in growth parameter and yield attributes when it increased up to 5.0 kg Zn ha-1. The highest yields were significantly receive in broad bed and furrow with 2.5 t ha-1 crop residue over narrow bed and furrow with 2.5 t ha-1 crop residue and flat bed with 2.5 t ha-1 crop residue, respectively with percent increment 16.33, 24.65, 11.47 and 7.16 in respect of biological yield, grain yield, stover yield and harvest index, respectively over control in pooled data. Seed inoculation with rhizobium produced significantly higher yields parameter over control and PSB with percent increment 5.37, 7.72, 3.84 and 2.34 in biological yield, grain yield, stover yield and harvest index, respectively over control. Application of 5.0 kg Zn ha-1 were produced significantly higher yields with percent increment 6.24, 7.25, 5.56 and 1.01 over control in respect of biological yield, grain yield, stover yield and harvest index, respectively. Therefore, broad bed and furrow with 2.5 t ha-1 crop residue and rhizobium inoculation with dose of 5.0 kg Zn ha-1 were significantly superior in respect to growth and yield attributes and yields of chickpea in present investigation.

The Growth Medium Affects the Viability of IPEC-J2 Animal Cell Line in the Presence of Probiotic Bacteria

Background: The IPEC-J2 cell line is frequently used as an in vitro model to study the bioactivity of live probiotics. However, lactic acid bacteria (LB) acidify the medium, and the impact of pH and lactic acid accumulation on cell viability seem to be underestimated. Methods: IPEC-J2 viability was assessed by neutral red and flow cytometry in the presence of eight probiotics at concentrations between 106 and 109 bacteria/mL in maintenance and buffered media. Results: It was shown that a high inoculation level led to higher cytotoxic effects on IPEC-J2 cells after 22 h of incubation and that viability losses were more related to a combination of low pH and lactic acid than to the probiotics themselves. Furthermore, with LB at 106 and 107 bacteria/mL, the addition of phosphates to the media significantly reduced the drop in the pH and preserved the IPEC-J2 viability between 100% and 69%, compared to a highly variable viability between 100% and 17.5% in the unbuffered media. Conclusions: Under certain in vitro conditions, probiotics can lead to the deterioration of animal cells, and pH neutralization is an essential parameter in the cell–probiotic system in order to preserve cell viability and to better evaluate the bioactive properties of live probiotics.

Effect of arbuscular mycorrhizal fungi on nutrients and heavy metals uptake by Pennisetum purpureum cv Mott in phytoremediation of gold mine tailings

Mercury composite and cyanidation are gold mining methods that are frequently used. The mercury composite method produces tailings containing heavy metals that can harm living organisms. Utilisation of tailings for the development of forage may be enhanced through arbuscular mycorrhizal fungi (AMF) inoculation to increase plant resistance, absorption of macro and micronutrients, and reduce levels of metal contaminants in the tailings. This study aimed to investigate the effect of arbuscular mycorrhizal fungi on nutrients and heavy metals uptake by &lt;em&gt;Pennisetum purpureum&lt;/em&gt; cv Mott in phytoremediation of gold mine tailings. Treatments consisted of four levels of AMF inoculation (0, 5, 10 and 15 g pot&lt;sup&gt;-1&lt;/sup&gt;) were arranged in a completely randomised design with five replications. Each pot contained 3 kg of tailings. The results showed that the best crude protein, crude fiber, crude fat, Ca, and P contents in the plant shoots was obtained by providing AMF inoculation of 15 g pot&lt;sup&gt;-1&lt;/sup&gt;. However, the treatment had no significant effect on dry weight, dry matter, and nitrogen-free extract. AMF significantly increased the uptake of heavy metals by the plant roots. The treatments did not significantly affect Pb uptake by plant roots and shoots and Hg uptake by plant shoots. AMF treatments significantly reduced the translocation factor (TF) value for Hg, bioconcentration factor (BCF) values for Cd and Pb, and removal efficiency (RE) values for Cd and Pb. AMF could effectively increase nutrient absorption in the plant shoots, reduce Cd, Hg, and Pb translocation in plant shoots, and reduce Cd, Hg, and Pb in the tailings.