Inoculated Research Articles

Integrating multiregulatory analysis reveals the negative regulatory function of miR482a in the response of poplar to canker pathogen infection.

Canker disease caused by the bacterium Lonsdalea populi is one of the most destructive diseases affecting poplar stems. However, the detailed stress response mechanisms of poplar have not been widely characterized. To explore the diverse regulatory RNA landscape and the function of key regulators in poplar subjected to L. populi stress, we integrated time-course experiment with mock-inoculation (CK) and inoculation (IN) with L. populi at the first, third, and sixth day (IN1, IN3, IN6) on Populus × euramericana cv. '74/76' (107), small RNA-seq, whole transcriptome-wide analysis, degradome analysis and transgenic experiments. A total of 98 differentially expressed (DE) miRNA, 17 974 DEmRNA, and 807 DElncRNA were identified in poplar infected by L. populi, presenting dynamic changes over the infection course. Regulatory networks among RNAs were further constructed. Notably, a network centered on ptc-miR482a in CK-vs-IN3 contained most DEGs. We show that miR482a and miR1448 are located in one transcript as a polycistron. Overexpression of pre-miR482a-miR1448 (OX482-1448) and pre-miR482a (OX482) increased poplar susceptibility to canker pathogen with reduced accumulation of reactive oxygen species, while the suppression of miR482a (STTM482) conferred poplar disease resistance. PHA7 was validated as the target of miR482a with degradome sequencing and tobacco transient co-transformation, its expression being downregulated in OX482-1448 and OX482 lines. Additionally, a series of phasiRNAs were triggered by miR482a targeting PHA7, forming regulatory cascades with more RLP, NBS-LRR, and PK genes, further verifying the defense function of miR482a. These findings provide insights for understanding the roles of ncRNAs and regulatory networks involved in poplar immunity.

PSX-27 Growth performance and immune response dynamics in weaned lambs with subclinical Haemonchus contortus parasite infection

Abstract Parasite infections, particularly from Haemonchus contortus, hinder sheep production in Canada by reducing weight gain and immunity. Current diagnosis and treatment methods rely on observing symptoms and the blind administration of anthelmintic drugs, resulting in escalating costs and the development of drug-resistant parasites. This study aimed to identify the exact time-point when subclinical H. contortus parasite infection affects growth performance and triggers an immune response. A total of 60 Rideau Arcott ewe lambs (~7-8 mo old), confirmed to be parasite-free, were randomly assigned in a 2 x 2 factorial design assessing inoculation treatment (TRT): oral inoculation (IN) with a solution containing 5,000 L3 strain of H. contortus or sham inoculation (CT) using water solution as control; and drench (DRENCH) administration: oral drench (D), 28-days post-inoculation, with a combination of 0.08% ivermectin solution (200 mcg per kg of BW; Durvet®) and 10% oral suspension of fenbendazole (2.5 mL per 50 kg of BW; Safe-guard®), or water solution (non-drenched, ND), resulting in four treatment groups: IN-D, IN-ND, CT-D, and CT-ND (n=15/treatment). Lambs were equally assigned to four feedlot pens (two replicates/treatment) for 57 days (d) with ad libitum access to a grain-based total mixed ration and water. Animals were individually weighed (BW) and blood sampled on days -1 and 0 (baseline), as well as on days 7, 21, 28, 35, 42, 56, and 57 post-inoculations (DAY); average daily gain (ADG) was calculated for the entire experimental period. White blood cells count (WBC) was used as an indicator of infection and red blood cell count (RBC) as indicator of anemia. Data was analyzed using a mixed-effects model including TRT, DRENCH, DAY (and their interactions) as fixed effects. Overall, CT lambs were heavier (TRT effect; P < 0.05) and had a tendency of greater (TRT effect; P = 0.08) ADG than IN lambs. The IN lambs had a tendency of greater (TRT*DAY; P = 0.07) WBC on d 7 than CT lambs. Additionally, IN-ND had lower (TRT*DRENCH; P < 0.05) WBC than IN-D lambs (1.9 and 2.0 ± 0.042 x 109/L, respectively). A significant (TRT*DRENCH*DAY; P < 0.01) interaction was found on RBC. The CT-D had lower (P < 0.05) RBC on d 7 and greater (P < 0.05) on d 28, 35, 42, and 57 than IN-ND as well as on d 28 than IN-D. Furthermore, CT-ND had greater (P < 0.05) RBC on d 21, 28 and 35 than IN-D and from d 21 to 57 than IN-ND. The IN-D had greater (P < 0.01) RBC from d 35 to 57 than IN-ND. These findings suggest compromised growth in both drenched and non-drenched infected groups. Infection slightly peaked in the infected lambs on d 7 post-inoculation, but drench administration was effective at mitigating overall infection by day 21 and the anemia by day 35.

Effect of Chemical and Microbial Additives on Fermentation Profile, Chemical Composition, and Microbial Populations of Whole-Plant Soybean Silage

This study evaluated the effects of two chemical additives or a microbial inoculant on chemical composition and DM losses in whole-plant soybean silage. One-hundred and twenty mini-silos were used in a completely randomized design experiment with the following treatments: water without chloride (control, CON); a microbial inoculant (INO); a chemical additive containing 35–45% formic acid (FA type); and another chemical additive containing 50–60% propionic acid (PA type). Data were analyzed using mixed models of SAS, and treatment differences were evaluated by the following orthogonal contrasts: C1 = CON vs. additives (INO + FA type + PA type); C2 = INO vs. chemical additives (FA type + PA type); and C3 = PA type vs. FA type. Silage pH and ammonia nitrogen concentration were decreased, and concentrations of lactic acid and acetic acid were increased with additives. Counts of lactic acid bacteria were higher in silages with INO than with chemical additives. DM recovery increased with FA type and PA type. Additives increased DM and CP concentrations. Silage A-fraction proportion was greater with additives. Additives, particularly FA type and PA type, improved chemical composition and fermentative profile and reduced undigestible proportions of protein in whole-plant soybean silage. Chemical additives were more effective in reducing silage DM losses than INO.

Effects of straw-returning combined with application of microbial inoculants on soil aggregates and related nutrients

We analyzed the particle size distribution of soil aggregates in 0-20 and 20-40 cm soil layers of rice-wheat rotation field based on a field plot test with two treatments, conventional straw returning (CK) and straw returning with the addition of straw decomposition promoting microbial inoculants (IT). We evaluated the water stability indices of soil aggregates (the number of soil water stable large aggregates R0.25, the average weight diameter MWD, and the geometric average diameter GMD), and measured the contents of soil organic carbon (SOC), total nitrogen (TN), and total phosphorus (TP) in the soil aggregates of ＜0.053, 0.053-0.25, 0.25-1, ＞1 mm. The results showed that: 1) The number of aggregates ＜0.053, 0.053-0.25, ＞0.25 mm in the 0-20 and 20-40 cm soil layers under IT decreased by 10.0% and 6.8%, increased by 3.0% and 5.7%, and 17.9% and 26.1% compared with CK, respectively. IT effectively increased R0.25, MWD, and GMD by 26.4%, 20.0%, 18.2% and 18.2%, 10.5%, 10.0% in 0-20 and 20-40 cm soil, respectively. 2) Compared to CK, the TP content of 0.25-1 mm aggregates in 0-20 and 20-40 cm soil under IT was significantly increased by 40.3% and 37.5%, respectively, without difference in TN and SOC contents. There was no significant difference in nutrient contents of the other aggregates between the treatments. The contents of SOC and TN in large aggregates (>0.25 mm) were higher than those in silty aggregates (<0.053 mm). Compared to CK, the cumulative contribution rates of SOC, TN and TP of <0.053 mm aggregates under IT were decreased in two soil layers. There was no significant difference in the nutrient cumulative contribution rates of 0.053-0.25 mm aggregates between treatments. The cumulative contribution rates of SOC, TN, and TP of large aggregates (>0.25 mm) under IT were 32.1%, 19.6%, 52.8% and 22.8%, 11.8%, 42.9% higher than those under CK in 0-20 and 20-40 cm soils, respectively. 3) The number of <0.053 mm aggregates was significantly negatively correlated with SOC and TP contents, while that of 0.053-0.25 mm aggregates was negatively correlated with nutrient content. The number of large aggregates (>0.25 mm) were significantly positively correlated with SOC, TN, and TP contents. In conclusion, straw returning with microbial-inoculant addition could promote the formation of soil macroaggregates (>0.25 mm), and improve the water stability of soil aggregates, increasing nutrient contents in soil macroaggregates, with the nutrients transferring from silty aggregates to macroaggregates.

Digestates from the co-digestion of cattle rumen and manure improve the methane potential of maize silage

The utilization of lignocellulosic biomasses for biogas production can slow down the biogas generation process. This study aimed to develop an effective inoculant composed of cattle manure rumen for the anaerobic digestion of maize silage. Four different proportions of cattle manure and rumen were tested as inoculants (IN) in this biomass. Biogas characterization, methane production efficiency, and the composition of the microbial community (both in the inoculants and reactors) were evaluated. The addition of IN1 (100 % cattle rumen) was unable to degrade the silage into methane (0.0 L kgVS−1). The final volume of biogas and methane produced by silage inoculated with IN3 (169.6 L kgVS−1) and IN4 (150.8 L kgVS−1) was higher. Analysis of the Archaeal communities in the silage and IN treatments revealed that the number of Amplicon Sequence Variants (ASVs) associated with Bathyarchaeia, Methanosarcinaceae, and Methanomassiliicoccaceae increased with an increase in the content of cattle manure in the inoculants. The findings support the promising potential of biogas and methane production from maize silage.

Effect of lignosulfonates on the dry matter loss, nutritional value, and microbial counts of high moisture alfalfa silage

In this experiment, we evaluated the effects of sodium lignosulfonate (NaL) and magnesium lignosulfonate (MgL) applied independently at 0, 5, 10, and 15 (g/kg, fresh weight basis) and an inoculant (INO) on high-moisture (DM: 219 g/kg, fresh weight basis) alfalfa (Medicago sativa L.) silage nutrient preservation. The INO consisted of Pediococcus pentosaceus and Lactobacillus plantarum applied at a rate of 4.95 and 4.00 log cfu/g (fresh weight basis), respectively. Data were analyzed as a randomized complete block design (RCBD; 5 blocks) and linear and quadratic polynomial contrasts were used to determine dose rate effects for NaL and MgL and orthogonal contrasts for INO effects. At d 0, increasing NaL dose from 0 to 15 g/kg increased linearly DM (219–227 g/kg) and decreased linearly mold counts (4.48–3.62 log cfu/g; fresh weight basis). No treatment effects were observed on lactic acid bacteria counts (7.04 log cfu/g; fresh weight basis), water soluble carbohydrates (61.5), and aNDF (432 g/kg of DM). After 229 d of ensiling, both MgL and INO increased DM loss (x¯= 13.7 and 13.7 vs 11.3% of DM) due to a lower production of lactic acid (x¯= 75.5 and 78.3 vs 92.3 g/kg of DM, respectively) which resulted in a higher pH relative to untreated silage (x¯= 4.41 and 4.46 vs 4.33; respectively). Increasing doses from 0 (6.42) to 15 g/kg for both NaL (6.90) and MgL (7.06 log cfu/g; fresh weight basis) resulted in linear and quadratic increases of lactic acid bacteria counts, respectively. These counts were also higher in INO treated silages (6.98 log cfu/g; fresh weight basis), relative to untreated silage. Furthermore, both NaL and MgL decreased in vitro ruminal total volatile fatty acid concentration (x = 87.0 and 93.1 vs 97.1 mM; respectively), compared with the untreated silage. The greater acidification in untreated silage prevented additives tested from reducing the generation of NH3-N (x¯= 110 g/kg of N) during ensiling. Overall, none of the additives tested improved the preservation of high-moisture alfalfa silage nutrients beyond what was observed in untreated.

Snaplage with microbial inoculant or organic acids has altered fermentative losses, microorganism counts, starch content and improves feed intake, digestibility and modulates ruminal fermentation in lambs

ABSTRACT Two experiments were performed to evaluate snaplage ensiling additives and determine the effects of feeding snaplage to lambs. In Exp. 1, grains, ears and husk from maize harvested at the R4 phenological stage were ensiled (65.3% DM content) in 30 min-silos in a completely randomised design with the following treatments: control (CON); microbial inoculant (INO), (Lactobacillus plantarum and Propionibacterium acidipropionici) at 4 g/ton of ensiled material; and organic acids (OA), a blend of organic acids at 2 g/kg of ensiled material. In Exp. 2, nine lambs were used in a 3 × 3 Latin square design. Lambs were randomly assigned to the following diets: control (CON); rehydrated maize silage (RHC), diet containing 380 g/kg RHC; or snaplage (SNAP), diet containing 770 g/kg of SNAP. Snaplage with OA exhibited greater DM recovery, lactic acid bacteria count and DM content than other treatments. Snaplage with INO had the lowest counts of aerobic bacteria and mould. Lambs-fed SNAP had greater nutrient intake and digestibility, ruminal short-chain fatty acid concentrations, and N retention compared to other groups. Incorporating OA to snaplage decreased DM losses and improved microbiological quality and chemical composition. Feeding snaplage to lambs may improve DM intake and apparent digestibility.

Effect of dry matter content on the microbial community and on the effectiveness of a microbial inoculant to improve the aerobic stability of corn silage

Silage inoculants are commonly used as a tool to improve the fermentation and aerobic stability of corn silage fed to dairy cows. However, their effectiveness can be inconsistent. Our objective was to determine the effect of the dry matter (DM) content of freshly chopped whole-plant corn on its microbial community as affected by an inoculant containing Lentilactobacillus hilgardii, Lentilactobacillus buchneri, and Pediococcus pentosaceus on improving the aerobic stability of silage. Whole-plant corn was harvested at low (31.80%, LDM), medium (33.32%, MDM), or high (39.44%, HDM) DM content and treated with nothing (CTR) or an inoculant (INO) containing L. hilgardii CNCM I-4785 at 150,000 cfu/g fresh forage, L. buchneri NCIMB 40788 at 150,000 cfu/g fresh forage, P. pentosaceus NCIMB12455 at 100,000 cfu/g of fresh forage, β-glucanase (5,750 IU/g), and xylanase (30,000 IU/g) and ensiled for 20 and 60 d. Data were analyzed as a completely randomized design in a 3-by-2 factorial arrangement of treatments. Fresh LDM forage had a higher concentration of reducing sugars, a less rich, diverse, and even bacterial community, and greater relative abundance of Saccharomycetales than MDM and HDM forages. Silages at 20 and 60 d, inoculated LDM had a more modest proliferation of culturable lactic acid bacteria than inoculated MDM. At 20 d, regardless of treatment, LDM had greater concentrations of lactic and acetic acids. Also at 20 d, LDM had lower numbers of culturable yeasts but greater relative abundance of Enterobacteriaceae than MDM and HDM. For silage at 20 d, HDM silage was more aerobically stable than LDM and MDM and inoculation improved aerobic stability 1.8-fold compared with CTR. For silage at 60 d, there was an interaction between DM content and inoculation. The improvements in stability by inoculation, compared with CTR, were greater in MDM (261 vs. 41 h) and HDM (320 vs. 66 h) silages than in LDM (85 vs. 46 h). The lower DM content and possible slower pH decline in LDM might have facilitated the development of undesirable bacteria and coupled with its greater concentration of reducing sugars and lactic and acetic acids, which are substrates for aerobic microorganisms, might explain the more modest improvements in aerobic stability from inoculation in LDM compared with MDM and HDM. Our findings suggest that the DM content of whole-plant corn affected its epiphytic microbial community and the effectiveness of the inoculant, which improved aerobic stability at all DM but to a greater extent in HDM and MDM than in LDM, especially after 60 d of ensiling.

Olive Mill Waste-Based Anaerobic Digestion as a Source of Local Renewable Energy and Nutrients

This study focused on what combination of anaerobic digestion (AD) temperature (ambient, mesophilic, and thermophilic) and olive mill waste (OMW) to dairy manure (DM) ratio mixture delivers the desired renewable energy and digestate qualities when using AD as olive mill waste treatment. OMW is widespread in the local environment in the North Sinai region, Egypt, which causes many environmental hazards if left without proper treatment. Three different mixtures consisting of OMW, dairy manure (DM), and inoculum (IN) were incubated under ambient, mesophilic, and thermophilic conditions for 45 days. The results showed that mixture B (2:1:2, OMW:DM:IN) at 55 °C produced more methane than at 35 °C and ambient temperature by 40% and 252%, respectively. Another aim of this study was to investigate the effects of the different concentrations of the digestate taken from each mixture on faba bean growth. The results showed that the maximum fresh weight values of the shoot system were observed at 10% and 15% for mixture B at ambient temperature. The best concentration value for the highest root elongation rate is a 5% addition of digestate mixture A at 55 °C, compared with other treatments.

In vitro Inoculation of Fresh or Frozen Rumen Fluid Distinguishes Contrasting Microbial Communities and Fermentation Induced by Increasing Forage to Concentrate Ratio.

In vitro rumen batch culture is a technology to simulate rumen fermentation by inoculating microorganisms from rumen fluids. Although inocula (INO) are commonly derived from fresh rumen fluids, frozen rumen fluids are also employed for the advantages of storing, transporting, and preserving rumen microorganisms. The effects of frozen INO on microbial fermentation and community may be interfered with by substrate type, which has not been reported. This study was designed to test whether rumen fluid treatments (i.e., fresh and frozen) could interact with incubated substrates. A complete block design with fractional arrangement treatment was used to investigate the effects of INO (fresh or frozen rumen fluids) and concentrate-to-forage ratios (C/F, 1:4 or 1:1) on rumen fermentation and microbial community. The effects of increasing C/F were typical, including increased dry matter (DM) degradation and total volatile fatty acids (VFA) concentration (P < 0.001), and decreased acetate to propionate ratio (P = 0.01) and bacterial diversity of richness and evenness (P ≤ 0.005) with especially higher fermentative bacteria such as genus Rikenellaceae_RC, F082, Prevotella, Bacteroidales_BS11, Muribaculaceaege, and Christensenellaceae_R-7 (P ≤ 0.04). Although frozen INO decreased (P < 0.001) DM degradation and altered rumen fermentation with lower (P ≤ 0.01) acetate to propionate ratio and molar proportion of butyrate than fresh INO, typical effects of C/F were independent of INO, as indicated by insignificant INO × C/F interaction on substrate degradation, VFA profile and bacterial community (P ≥ 0.20). In summary, the effect of C/F on fermentation and bacterial diversity is not interfered with by INO type, and frozen INO can be used to distinguish the effect of starch content.

Innovations in market crop waste compost production: Use of black soldier fly larvae and biochar

Purpose Compost production technology and use are known among Ghanaians farmers. However, the long composting period averaging three months has had adverse effect on adoption. The black soldier fly (Hermetia illucens) larvae (BSFL) feed voraciously and breaks down organic wastes fast. Addition of rice husk biochar (RHB) neutralises acidity and aerates the compost. Combined application of BSFL and RHB to feedstocks could therefore reduce composting period and improve compost quality. Method Two studies were conducted with market crop waste (MCW) as feedstock. Firstly, feedstocks at two particle sizes (x 10 mm), with and without BSFL were evaluated to determine degradability, chemical content of the degraded residue and to select appropriate feedstock size ideal for composting. Secondly, the selected feedstocks size of between 5 mm and 10 mm were inoculated with or without BSFL and RHB of 0%, 5% 10% and 15% added. Composting trials were conducted in barrels inclined at 30o to facilitate drainage. Physicochemical and biological parameters of feedstock were monitored until maturity. Results Degradability of MCW by BSFL was feedstock specific rather than feedstock size. Inoculation of BSFL and biochar addition reduced composting period from 76 to 45 days. Biochar addition at 15% increased P availability to 1882 mg kg-1 but reduced total N to 10.5 g kg-1. E coli levels decreased in the BSFL-biochar composts to acceptable limits. Conclusion Composting MCW with BSFL and biochar reduced composting period and improved compost quality.

Microbiological integration for qualitative improvement of vermicompost

Purpose This study was carried out to assess the effect of integrating microbiological fortification with vermicomposting process on biofertilizing quality of vermicompost as well as improvement of the availability of nitrogen and phosphorus in the product. Method A mixture of cow dung and vegetable market waste (1:1) was used for vermicomposting with Eisenia foetida as the decomposer earthworm @ 10 no.of worm kg-1 substrate. Nitrogen fixing and phosphate solubilizing bacteria (NFB and PSB)Azotobacter chroococcum and Pseudomonas fluorescens were integrated with this composting process at varying doses viz. 0,5 and 10 g kg-1 substrate under different combinations. Changes in the population of these two bio-fertilizing microorganisms and the availability of relevant nutrients in the substrates were monitored periodically to assess the behaviors of these microorganisms and their effects on the produced vermicompost. Results All the inoculations resulted in substantial increments in population of both NFB and PSB over the control. However, the increments were more prominent for NFB than the PSB. Significant increments in the amount of mineralized nitrogen and solubilized phosphate over the control were observed in almost all the treatment combinations. Conclusion Integration of nitrogen fixing and phosphate solubilizing bacteria with vermicomposting process resulted in substantial enrichment of the product. This benefit was observed not only in terms of increased availability of the two major plant nutrients of concern viz. N and P, but also in significant improvement in the population of the inoculated microorganisms turning the product into a potential source of bio-fertilizers.

Multi-Level Characterization of Microbial Consortia Involved in the Biodeterioration of Wooden and Stone Romanian Heritage Churches

The 17th to 19th century wooden and stone churches are an iconic symbol of Romanian national heritage. The present study investigates by qualitative and quantitative methods the microbial communities from the biodeteriorated surfaces of wooden and stone monument churches included in the cultural heritage list of local or national importance. From a total of twelve monuments, samples were taken with cotton sterile swabs, inoculated on specific culture and identified by classical, automated and molecular methods. A total of 133 strains belonging to Ascomycota phylum were identified and confirmed at species level from the wooden churches, amongst which, Penicillium spp. strains (mostly P. corylophylum, P. chrysogenum) were the most frequent, followed by Alternaria alternata and species of Trichoderma, Aspergillus, Rhizopus, Mucor, and Fusarium genera. From the stone churches a total of 100 strains belonging to Aspergillus, Alternaria, Mucor, Penicillium, Aspergillus, Trichoderma, Fusarium and Rhizopus genera were isolated. A total of 55 bacterial strains were isolated and identified as Bacillus, Artrobacter and Pseudomonas species. The microbial load of the samples ranged between 2.18x107 and 3x105 CFU/mL. A very small number of fungal strains (6/77) isolated from wooden churches (mostly A. alternata, followed by P. corylophilum and one Cladosporium spp. strain) and from stone churches (5/55) (mostly A. alternata, followed by A. versicolor, A. nidulans strain) were involved in biofilm formation. The results of this study can help to improve understanding of the microbial deterioration of Romanian heritage churches and allow more reliable decontamination, conservation and preservation tools to be defined.

Diversity and distribution of arbuscular mycorrhizal fungi in maize (Zea mays) cropping fields in South Kivu, Democratic Republic of Congo

Arbuscular Mycorhizal Fungi (AMF) occur naturally in agroecosystems and interact symbiotically with crops, facilitating nutrition. This study aimed at assessing the occurrence, abundance and diversity of AMF communities in the maize cropping system and their relation to soils properties in two agroecological zones in South Kivu, eastern DR Congo. Soil samples were collected from eight sites, with 4 sites in highland at 1400 to 2000 m altitude above sea level (in Katana, Kavumu, Mulamba and Mugogo) and four in the lowland at <1000 m (in Luberizi, Bwegera, Luvungi and Kamaniola). Spores were extracted from the field soils, morphologically identified and counted. AMF spores occurrence, abundances, species richness, and diversity were determined. A total of 38 AMF morphotypes distributed in 11 genera were obtained with the majority being from Gigasporaceae, Acaulosporaceae, and Glomeraceae families. This is the first report on the occurrence of these species in the eastern of DR Congo. Acaulospopra excavata, Acaulospopra bireticulata, Densitscutata erythropa, Funneliformis mosseae and Scutellospora pellucida were ubiquitous in all the agroecologies. Spores densities were higher in the highland with the highest recorded in Mulamba. Soil pH and phosphorus content influenced AMF distribution. The many different ubiquitous species indicate adaptation to a wide range of physicochemical environments and could reduce the cost of AMF inoculants production for the region. Maize agroecosystems are rich in AMF diversity and selection of appropriate fungal species from the Gigasporaceae, Acaulosporaceae and Glomeraceae as biofertilizer could contribute in improving crops production. Key words: Arbuscular mycorrhizal fungi, diversity, occurence, agroecological zones, maize, South Kivu.

Production and Comparative Physicochemical Analysis of Vinegar from Locally Grown Fruits in Nigeria and Industrial Produced Vinegar

Vinegar is the product made from the conversion of ethyl alcohol to acetic acid by a genus of bacteria, Acetobacter. The major aim of the work is to produce a vinegar with locally made fruits and compare it with the industrial produced vinegar to ascertain if it can be used to substitute the industrial produced vinegar and to also determine if there are presence of acetic acid bacteria present in the vinegar which can be used as inoculants for future fermentation the sensory attributes were also analysed to determine the consumer acceptability of the vinegar produced. This work involves production of vinegar from locally produced fruits in Nigeria by homemade scale process and then analyzing the vinegar to determine the physicochemical components of the vinegar as well as identifying the acetic acid bacteria present in the vinegar and its sensory attributes. The vinegar produced was analysed for its pH, relative density and titrable acidity and from the result, there was no significant difference in the pH and relative density of the vinegars. The titrable acidity of the industrial produced vinegar is higher than the locally produced vinegars analysed. The microbial isolation of the vinegar for safety assessment gave no growths in the media used for the analysis which means that there are no pathogenic organisms like mould or coliform which can cause diseases to the consumers. The major organism isolated was the Acetobacter spp. although the culture independent methods should be carried out to determine other non culturable organisms present in the vinegar. The sensory evaluation shows that the locally produced was well accepted from the scores on the result presented. Generally, the result of this work shows that indeed vinegar of good quality can be produced locally using locally grown fruits but optimization of the process should be carried out to improve yield.

Inoculating against ignorance.

Inoculating against ignorance.

Degradation of lignocellulosic content of rice straw using aerobic cellulolytic bacteria isolated from forest soil of Bangladesh

The present research work was conducted to enumerate and isolate aerobic cellulolytic bacteria from forest soil using carboxymethyl cellulose (CMC) as substrate. The identified isolate was further tested as potent candidate to improve the nutritional value of rice straw. The forest soil was loaded with substantial amount (2.45×107) of cellulolytic bacteria. A total of 10 cellulose degrading bacteria (CDB) were isolated, identified and monitored for their cellulolytic activity. The isolate having the highest cellulolytic index (2.5) was identified as Bacillus subtilis targeting the 16S rRNA gene which was labeled as B. subtilis strain CDB7 and investigated for nutritional improvement of rice straw. Solid state fermentation of each group was carried out at 37°C for a period of 0, 3 and 6 days in 0 (no bacterial inoculum), 1, 5 and 10% inoculum group. There were a tendency of reduction in Acid Detergent Fiber (ADF), Neutral Detergent Fiber (NDF) and lignin at concentrations of 1, 5 and 10% inoculums at every interval of fermentation. Highest reduction of ADF (4.8%), NDF (10.78%) and lignin (37.6%) were observed after 6 days of fermentation at 10, 10 and 5% inoculum group, respectively. The crude protein content was increased (5.3 ± 0.4 to 6.4± 0.3) in 10% inoculum group. Taken together, the identified isolate could be a potent candidate to degrade lignocellulosic content through breaking of lignin-cellulose bondage and to improve the nutritional value of rice straw. Key words: Rice straw, Bacillus subtilis strain CDB7, solid state fermentation, acid detergent fiber (ADF), neutral detergent fiber (NDF), lignin.

Adana ve Mersin nar bahçelerinde derim öncesi meyve çürüklük patojenleri ve farklı zamanlardaki inokulasyonların hastalık oluşumuna etkisi

Derim öncesi meyve çürüklüğü nar bahçelerinde önemli bir problemdir ve ciddi ürün kayıplarına neden olmaktadır. Bu çalışmanın amacı Adana ve Mersin illerindeki nar bahçelerinde derim öncesi meyve çürüklüğünün yaygınlığını saptamak, bununla ilişkili fungal patojenleri belirlemek ve farklı zamanlarda yapılan inokulasyonların hastalık oluşumuna etkisini ortaya koymaktır. 2018 yılında toplam 39 bahçede sürvey yapılmış ve simptomatik meyve örnekleri alınmıştır. Fungal patojenler standart mikolojik prosedürlere göre izole edilmiş ve klasik ve moleküler tekniklerle tanılanmıştır. Fungal patojenler bahçe koşullarında 2 farklı zamanda (çiçeklenme-meyve tutumu ve meyve büyümesi dönemleri) inokule edilmiş ve sırasıyla meyve dökümü ile kaliks çürüklüğü görülen meyve oranları kaydedilmiştir. Sonuçlara göre, Adana ve Mersin’deki derim öncesi meyve çürüklüğünün yaygınlık oranlarının sırasıyla %70.6 ve %22.7 olduğu bulunmuştur. Bu illerdeki ortalama hastalık çıkışı yine aynı sırayla %5.0 ve %1.1 olarak hesaplanmıştır. Simptomatik meyvelerden Alternaria spp., Aspergillus niger, Aureobasidium pullulans, Botrytis cinerea, Colletotrichum spp., Diaporthe ambigua, Nigrospora oryzae, Penicillium spp. ve Talaromyces spp. türlerine ait izolatlar elde edilmiştir. Bu türlerin bazıları (Alternaria alternata, Aspergillus niger, Botrytis cinerea, Colletotrichum gloeosporioides, Coniella granati, Diaporthe ambigua ve Penicillium mallochii), çiçekli meyveciklere inokule edildiklerinde, %87.5-100 oranında dökülmeye neden olmuş, ancak saf su püskürtülen kontrolde dökülme oranları %60-70 arasında gerçekleşmiştir. Buna karşın meyveler yarı büyüklüğe ulaştığında sadece Coniella granati ve Diaporthe ambigua inokulasyonları kaliks çürüklüğüne neden olmuş, diğer türler meyveleri çürütememiştir. Bu sonuçlar, inokule edilen türlerin çiçek ve meyvecikler üzerinde patojenik olduğunu ve bu kısımların dökülmesine yol açtığını göstermiştir. Diaporthe ambigua’nın şimdiye kadar nar meyve çürüklüğüyle ilişkili olduğuna dair bir kayda rastlanmazken bu çalışma ile meyvelerde saldırgan bir tür olduğu ortaya konmuştur.

Potensi Fungisida Perlakuan Benih terhadap Perenosclerospora sp. penyebab Penyakit Bulai Jagung

Jamur patogen Perenosclerospora sp. merupakan penyakit penting pada tanaman jagung. Dampak yang ditimbulkan dapat menimbulkan kerugian hingga 100%. Penelitian ini bertujuan untuk mengetahui pengaruh bahan aktif fungisida sebagai perlakuan benih dalam mengendalikan penyakit bulai (Perenosclerospora sp.) pada tanaman jagung. Penelitian dilaksanakan dengan diawali menanam tanaman jagung yang diinokulasi bulai sebagai tanaman spreader (sumber inokulum alami) dan pembatas antar perlakuan. Selanjutnya saat tanaman border sudah terinfeksi bulai baru ditanam benih perlakuan yang sudah dicampur dengan bahan aktif fungisida. Penelitian disusun dalam rancangan acak lengkap dengan delapan perlakuan dengan tiga ulangan. Perlakuan terdiri atas kontrol tanpa fungisida (P0), dimetomorf (P1), dimetomorf + piraklostrobin (P2), fenamidon (P3), fenamidon dan propamokarb hodroklorida (P4), metalaksil (P5), mefenoksam (P6), dan fluopikolid+propineb (P7). Hasil penelitian menunjukkan perlakuan kontrol lebih bagus dalam meningkatkan daya kecambah dibandingkan dengan menggunakan fungisida. Benih jagung yang diberikan perlakukan bahan aktif dimetomorf + piraklostrobin memiliki potensi yang paling tinggi dalam menekan keterjadian penyakit bulai pada jagung manis dan mampu meningkatkan pertumbuhan tinggi tanaman jagung yang dibandingkan perlakuan lainnya.

Physiological changes of Larix principis-rupprechtii seedlings inoculated with Trichoderma spp. under drought stress.

Tree regeneration is significantly affected by water deficiency, which could be alleviated by the inoculation of Trichoderma spp. In this study, the effects of rhizosphere inoculation with Trichoderma spp. on the antioxidant system and osmotic substances in the seedlings of Larix principis-rupprechtii was investigated under experimental drought stresses, with the relative water content 50%-60%, 35%-50%, and 20%-35% as the light, moderate and severe drought stress, respectively. The results showed that, with increasing drought stress, the activities of protective enzymes, SOD and POD in the needles of seedlings were significantly promoted, the contents of MDA and proline were remarkably increased, whereas the contents of soluble protein and starch showed a decreasing trend. Inoculation with Trichoderma spp. promoted much stronger increase in the activities of SOD and POD in the needles, yet the MDA content showed a lower level of increase. Under moderate drought, the activity of POD in the seedling inoculated with Trichoderma spp. was 1.8 folds of that in the control, while the content of MDA was 62.9% of the control. The contents of proline, soluble protein, soluble sugar, and starch in the inoculated seedlings were higher than that in the uninoculated seedlings. Drought-resistance of L. principis-rupprechtii seedlings inoculated by Trichoderma spp. was significantly enhanced due to the augmenting regulation of antioxidant system and osmotic adjustment substances. Therefore, under the trend of warming and drying in North China, the technique of rhizosphere inoculation of Trichoderma spp. could be popularized for forest plantation or understory tending.