The role and mechanisms of microbes in dichlorodiphenyltrichloroethane (DDT) and its residues bioremediation.

Environmental contamination with dichlorodiphenyltrichloroethane (DDT) has sever effects on the ecosystem worldwide. DDT is a recalcitrant synthetic chemical with high toxicity and lipophilicity. It is also bioaccumulated in the food chain and causes genotoxic, estrogenic, carcinogenic, and mutagenic effects on aquatic organisms and humans. Microbial remediation mechanism and its enzymes are very important for removing DDT from environment. DDT and its main residues dichlorodiphenyldichloroethylene (DDE) and dichlorodiphenyldichloroethane (DDD) can biodegrade slowly in soil and water. To enhance this process, a number of strategies are proposed, such as bio-attenuation, biostimulation, bioaugmentation and the manipulation of environmental conditions to enhance the activity of microbial enzymes. The addition of organic matter and flooding of the soil enhance DDT degradation. Microbial candidates for DDT remediation include micro-algae, fungi and bacteria. This review provide brief information and recommendation on microbial DDT remediation and its mechanisms.

Host Specificity Controlled by PWL1 and PWL2 Effector Genes in the Finger Millet Blast Pathogen Magnaporthe oryzae in Eastern Africa.

Magnaporthe oryzae, a devastating pathogen of finger millet (Eleusine coracana), secretes effector molecules during infection to manipulate host immunity. This study determined the presence of avirulence effector genes PWL1 and PWL2 in 221 Eleusine blast isolates from eastern Africa. Most Ethiopian isolates carried both PWL1 and PWL2. Kenyan and Ugandan isolates largely lacked both genes, and Tanzanian isolates carried either PWL1 or lacked both. The roles of PWL1 and PWL2 towards pathogenicity on alternative chloridoid hosts, including weeping lovegrass (Eragrostis curvula), were also investigated. PWL1 and PWL2 were cloned from Ethiopian isolate E22 and were transformed separately into Ugandan isolate U34, which lacked both genes. Resulting transformants harboring either gene gained varying degrees of avirulence on Eragrostis curvula but remained virulent on finger millet. Strains carrying one or both PWL1 and PWL2 infected the chloridoid species Sporobolus phyllotrichus and Eleusine tristachya, indicating the absence of cognate resistance (R) genes for PWL1 and PWL2 in these species. Other chloridoid grasses, however, were fully resistant, regardless of the presence of one or both PWL1 and PWL2, suggesting the presence of effective R genes against PWL and other effectors. Partial resistance in some Eragrostis curvula accessions to some blast isolates lacking PWL1 and PWL2 also indicated the presence of other interactions between fungal avirulence (AVR) genes and host resistance (R) genes. Related chloridoid species thus harbor resistance genes that could be useful to improve finger millet for blast resistance. Conversely, loss of AVR genes in the fungus could expand its host range, as demonstrated by the susceptibility of Eragrostis curvula to finger millet blast isolates that had lost PWL1 and PWL2. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Biological management of coffee wilt disease (Fusarium xylarioides) using antagonistic Trichoderma isolates.

Coffee wilt disease (CWD) is a serious threat to the food security of small-scale farmers in Ethiopia, causing significant reductions in coffee yield. Currently, there are no effective control measures available against the causative agent of CWD, Fusarium xylarioides. The main objective of this study was therefore to develop, formulate, and evaluate a range of biofungicides against F. xylarioides, derived from Trichoderma species and tested under in vitro, greenhouse, and field conditions. In total, 175 Trichoderma isolates were screened as microbial biocontrol agents against F. xylarioides. The efficacy of two biofungicide formulations, wettable powder and water dispensable granules, were tested on the susceptible Geisha coffee variety in three different agro-ecological zones in southwestern Ethiopia over three years. The greenhouse experiments were set up using a complete block design, while in the field a randomized complete block design was used, with twice yearly applications of biofungicide. The test pathogen spore suspension was applied to the coffee seedlings by soil drenching, and the subsequent incidence and severity of CWD evaluated annually. The mycelial growth inhibition profiles of the Trichoderma isolates against F. xylarioides ranged from 44.5% to 84.8%. In vitro experiments revealed that T. asperelloides AU71, T. asperellum AU131 and T. longibrachiatum AU158 reduced the mycelial growth of F. xylarioides by over 80%. The greenhouse study indicated that wettable powder (WP) of T. asperellum AU131 had the highest biocontrol efficacy (84.3%), followed by T. longibrachiatum AU158 (77.9%) and T. asperelloides AU71 (71.2%); they also had a significant positive impact on plant growth. The pathogen-treated control plants had a disease severity index of 100% across all the field experiments, and of 76.7% in the greenhouse experiments. In comparison to untreated controls, the annual and cumulative disease incidence over the three years of the study period varied from 46.2 to 90%, 51.6 to 84.5%, and 58.2 to 91%, at the Teppi, Gera and Jimma field experimental locations. Overall, the greenhouse and field experiments and in vitro assays support the biocontrol potential of Trichoderma isolates, and T. asperellum AU131 and T. longibrachiatum AU158 in particular are recommended for the management of CWD under field conditions.

Isolation and Determination of Antibacterial Sensitivity Characteristics of Staphylococcus aureus from Lactating Cows in West Shewa Zone, Ethiopia.

Staphylococcus (S.) aureus is one of the etiologies of bovine mastitis, hindering milk production and productivity in dairy farms. This study was aimed at assessing the distribution of bovine mastitis and the isolation rate of S. aureus in milked cows of West Shewa Zone. The clinical mastitis was diagnosed by physical methods including observation and palpation, whereas the subclinical mastitis was tested by the California mastitis test (CMT). All of the cows tested for mastitis were aseptically sampled (teat-milk) for bacteriology. The bacterium was primarily identified based on colony characterization, catalase, coagulase tests, and Gram stain reaction. Finally, MALDI-TOF Biotyper confirmed the species. The antibacterial sensitivity characteristics of the isolates to different antibacterial drugs were tested by the disk diffusion method. The drugs were selected based on the frequent usage in veterinary medicine in the study area. By using particular primers, the presence of the resistance (mecA and blaZ), and thermonuclease (nuc) genes were determined by polymerase chain reaction (PCR). The data were analyzed by R statistical software. The associations between the dependent variables (prevalence of mastitis and S. aureus) and the explanatory variables were analysed by chi-square (χ 2) and logistic regression tests at a 95% confidence interval (CI). Accordingly, 258 lactating cows were examined, of which 97 (37.6%) were mastitis positive. Of these mastitis positive cows, 59 (60.8%) were subclinical and 38 (39.2%) were clinical. Among the 258 milk samples, 43 (16.7%) were positive for S. aureus. According to the results of the current investigation, subclinical mastitis was significantly more prevalent than clinical mastitis (p < 0.05). The disease was found varied with the lactation stage of the animal, milking with washed hand, udder washing before milking, and tick infestation of the teat. In comparison to animals from farms with lower number of lactating cows, the prevalence of the bacteria was significantly higher in animals managed in farms with large (OR = 12.58, 95% CI = 2.33-68.54, and p < 0.05) and medium (OR = 12.58, 95% CI = 2.33-68.54, and p < 0.05) population of lactating cows per herd. The isoation rate of the bacterium was also found significantly higher in tick-infested cows (OR = 27.69, 95% CI = 9.71-93.01, and p < 0.05) than tick free cows. The antibiogram tests revealed that the isolates resisted penicillin G and tetracycline group drugs (oxytetracycline and tetracycline). Moreover, the nuc gene was confirmed to be present in all of the examined isolates. However, they were not found harboring blaZ and mecA genes. We concluded that S. aureus is sustaining as a main causative agent of bovine mastitis, and they were resistant to the frequently used antibiotics in public and veterinary medicines in the study areas. Therefore, research-based interventions need to be taken in action to combat the pathogen.

Evaluation of finger millet (Eleusine coracana (L.) Gaertn.) in multi-environment trials using enhanced statistical models.

Spatial variation and genotype by environment (GxE) interaction are common in varietal selection field trials and pose a significant challenge for plant breeders when comparing the genetic potential of different varieties. Efficient statistical methods must be employed for the evaluation of finger millet breeding trials to accurately select superior varieties that contribute to agricultural productivity. The objective of this study was to improve selection strategies in finger millet breeding in Ethiopia through modeling of spatial field trends and the GxE interaction. A dataset of seven multi-environment trials (MET) conducted in randomized complete block design (RCBD) with two replications laid out in rectangle (row x column) arrays of plots was used in this study. The results revealed that, under the linear mixed model, the spatial and factor analytic (FA) models were efficient methods of data analysis for this study, and this was demonstrated with evidence of heritability measure. We found two clusters of correlated environments that helped to select superior and stable varieties through ranking average Best Linear Unbiased Predictors (BLUPs) within clusters. The first cluster was chosen because it contained a greater number of environments with high heritability. Based on this cluster, Bako-09, 203439, 203325, and 203347 were the top four varieties with relatively high yield performance and stability across correlated environments. Hence, scaling up the use of this efficient analysis method will improve the selection of superior finger millet varieties.

Isolation of Hexavalent chromium tolerant fungal species from urban vegetable farm soil and effluent waste in Addis Ababa& Rift valley, Ethiopia.

Hexavalent chromium is resistant to degradation and harmful toxic substance to environment and community health. Physicochemical treatment methods are demanding high cost, used large quantities of chemicals & energy, release large amount of secondary toxic degradants. Mycoremediation is an eco-friendly alternative treatment method. The main objective of this research is to isolate and characterize chrome (VI) tolerant fungi from farm soil & industry effluent for mycoremedation purpose. The screening and isolation of yeast was carried out on potato dextrose agar media. PDA and broth assay test for fungi tolerance to hexavalent chromium at different concentration, temperature and pH was evaluated. Fungi species was identified biochemically using Biolog Microstation depending on carbon utilization and chemical sensitivity test. The result revealed that 10 yeast species was identified with full ID from effluent waste and farm soil based on their probability ≥  75% and similarity index ≥  0.5 as well as their Cr (VI) tolerance ability up to 2500 ppm. These are Yarrowia lipolytica (100%, 0.7), Cryptococcus luteolus(100%, 0.64), Rhodotorula aurantiaca A(100%, 0.62), Ustilago maydis(100%, 0.58) Trichosporon beigelii B (100%, 0.51), Cryptococcus terreus A (100%, 0.62), Zygosaccharomyces bailii (98%, 0.65), Nadsoniafulvenscens (90%, 0.62), Schizoblastosporonstarkeyihenricii (89%, 0.56), Endomycopsis vivi (84%, 0.62), Rhodotorula pustula (Sim, 0.59). Two yeast species Yarrowia lipolytica and Nadsoniafulvenscens show the highest growth mean Optical density (OD) measure 0.74 ± 0.2 & 0.60 ± 0.2 respectively at pH 7 & 25 °C. The highest tolerance index (mm) was recorded by Schizoblastosporon starkey henricii 0.3067 ± 0.152. Cr (VI)-tolerance ability of these yeast strains used in the development of chromium-bioremediation technologies provide an alternative option for chromium sequestration after HPLC analysis& molecular characterization.

Plant growth-promoting rhizobacterial inoculation to improve growth, yield, and grain nutrient uptake of teff varieties.

Inoculation of plant growth-promoting rhizobacteria (PGPR) improves the growth, yield, and plant nutrient uptake, as well as rhizosphere fertility, without harming the environment and human health. This study aimed to examine the effect of either individual or consortium of PGP bacterial inoculation on the growth, yield, and grain nutrient uptake of teff varieties. Three potential PGPR strains (i.e., Pseudomonas fluorescens biotype G, Enterobacter cloacae ss disolvens, and Serratia marcescens ss marcescens) were used for this study. Field evaluation was carried out in RCBD with 5 treatments. Highly significant (P < 0.001) differences were observed among treatments for plant height (PH), panicle length (PL), number of the total spike (NTS), shoot dry weight (SDW), grain yield (GY), and straw yield (SY). There was also teff variety that significantly (P < 0.01) affects PL, SDW, and SY. However, the interaction effect of the two factors (treatment*variety) did not significantly influence teff agronomic traits and grain nutrient uptake. The highest PH (133.5 cm), PL (53.2), NTS (30.9), SDW (18.1 t/ha), SY (10.7 t/ha), and GY (2.7 t/ha) were observed on Dukem variety (Dz-01-974) inoculated with PGPR consortium. Wherein 2.2 fold increase was observed in grain yield per hectare over the control. Inoculation of PGPR consortium showed better performance in promoting plant growth, yield, and grain nutrient uptake of teff varieties compared with the individual PGP bacterial application, and PGPR consortium could be used as inoculants to enhance teff production and productivity.

In Vitro Evaluation of Extracellular Enzyme Activity and Its Biocontrol Efficacy of Bacterial Isolates from Pepper Plants for the Management of Phytophthora capsici.

Phytophthora capsici is one of the most devastating fungal pathogens, causing severe diseases that lead to economic loss in the pepper industry. As a result of the infections, the chemical approach is becoming more popular. Biological control, on the other hand, is better suited to controlling fungal pathogens. The biological control approach significantly reduces the problems associated with chemical applications while restoring natural environmental balance. As a result, the overall findings indicate that certain bacterial isolates play a beneficial role in lytic enzyme production and biocontrol activities against P. capsici. Bacterial isolates obtained from the pepper plants were screened for lytic enzyme and anti-oomycete activity against Phytophthora capsici in Ethiopia. Sixty bacterial isolates were isolated and tested against Phytophthora capsici. From these bacterial isolates, different inhibition zones and hydrolytic enzyme production were detected. Biochemical tests using an automated machine (MALDI-TOF, VITEK 2 compact and 16S rRNA) revealed that three of them, AAUSR23, AAULE41, and AAULE51, showed a high inhibition zone and high production of hydrolytic enzymes and were identified as Enterobacter cloacae (AAUSR23), Pseudomonas fluorescens (AAULE41), and undetermined (AAULE51). The effects of diffusable metabolite isolate AAULE51 has a 66.7% inhibition zone against Phytophthora capsici, followed by AAULE41 and AAUSR23, which have 59.7% and 14.1% inhibition zones, respectively. These bacterial isolates showed high production of hydrolytic enzymes like protease, cellulase, chitinase, and lipase (5-34 diameter of inhibition zone). As a result, the overall findings show that selected bacterial isolates play a beneficial role in lytic enzyme production and for their biocontrol activities against P. capsici.

Antifungal compounds, GC-MS analysis and toxicity assessment of methanolic extracts of Trichoderma species in an animal model.

Fungi of the genus Trichoderma have been marketed for the management of diseases of crops. However, some Trichoderma species may produce toxic secondary metabolites and it should receive due attention to ensure human safety. In this study, we investigated the in vitro antagonistic potential of T. asperellum AU131 and T. longibrachiatum AU158 as microbial biocontrol agents (MBCAs) against Fusarium xylarioides and the associated antagonistic mechanism with bioactive substances. Swiss albino mice were used to evaluate the in vivo toxicity and pathogenicity of T. asperellum AU131 and T. longibrachiatum AU158 methanolic extracts and spore suspensions, respectively, in a preliminary safety assessment for use as biofungicides. Gas Chromatography-Mass Spectrometry (GC-MS) was used to profile volatile organic metabolites (VOCs) present in the methanolic extracts. The agar diffusion assay of the methanolic extracts from both T. asperellum AU131 and T. longibrachiatum AU158 were effective at a concentration of 200 µg/mL (1×107 spores/mL), causing 62.5%, and 74.3% inhibition, respectively. A GC-MS analysis of methanolic extracts from both bioagents identified 23 VOCs which classified as alcohols, acids, sesquiterpenes, ketones and aromatic compounds. The oral administration of methanolic extracts and spore suspensions of each Trichoderma species to female Swiss albino mice over 14 days did not show any significant signs of toxicity, mortality or changes to body weight. It can be concluded that the tested spore suspensions and methanolic extracts were not pathogenic or toxic, respectively, when administered to Swiss albino mice at various doses.

Characterization of faba bean (Vicia faba L.) rhizosphere associating rhizobacteria against Botrytis fabae AAUBF-12 and their plant growth-promoting properties.

The rhizobacteria are known to protect plants from different pathogens acting as biocontrol agents and promote growth of plants. This study was conducted to isolate, screen and identify faba bean associating rhizobacteria for their antagonistic properties against Botrytis fabae AAUBF-12 and plant growth-promoting properties under in vitro conditions. In the dual culture assay, the isolates inhibited the mycelia growth of B. fabae AAUBF-12 (6-40 %) upon 3 days of incubation, and the inhibition increased to 9-43 %, 16-50 %, and 24-68 % after five, seven and 9 days of incubation, respectively. The inhibitory activity increased from 6 to 82 % using the culture filtrates of the isolates. Isolate AAUB95 displayed the highest mycelial inhibition (27 %) at 5 % concentration of culture filtrate, followed by AAUB146b that exhibited 21 % inhibition at the same concentration. AAUB146b and AAUB100 effectively inhibited B. fabae AAUBF-12 by 79 % and 80 % at 20 % concentrations of the culture filtrate. The qualitative study demonstrated 75 % of the isolates positive for protease and 60 % for lipase synthesis. Furthermore, the isolates that showed antagonistic activity against B. fabae AAUBF-12, produced IAA and ammonia with 65 % and 60 %, respectively. Moreover, 310-760 µg mL-1 and 200-620 µg mL-1 of tricalcium phosphate (TCP) was released on the 3rd and 6th days of incubation, respectively, due to rhizobacterial solubilization. Nevertheless, the Pearson's correlation analysis between pH and TCP solubilization revealed an inverse relationship (r = -.422∗∗). Based on 16S rRNA sequences analysis, isolate AAUB95, AAUB146b, AAUB100 and AAUB92 were identified as B. subtilis AAUB95, S. nematodiphila AAUB146b, B. tequilensis AAUB100 and B. subtilis AAUB92, respectively. Of the isolates, B. subtilis AAUB95 showed best antagonism of B. fabae AAUBF-12 with multiple plant growth-promoting properties.

Optimization of Culture Conditions and Production of Bio-Fungicides from Trichoderma Species under Solid-State Fermentation Using Mathematical Modeling.

Agro-industrial wastes suitable for economical and high mass production of novel Trichoderma species under solid-state fermentation were identified by optimizing the culture conditions using a mathematical model and evaluating the viability of the formulated bio-product. Fourteen inexpensive, locally available, organic substrates and cereals were examined using a one-factor-at-a-time experiment. The fungus colonized nearly all substrates after 21 days of incubation, although the degree of colonization and conidiation varied among the substrates. A mixture of wheat bran and white rice (2:1 w/w) was found to support maximum growth of T. asperellum AU131 (3.2 × 107 spores/g dry substrate) and T. longibrachiatum AU158 (3.5 × 107 spores/g dry substrate). Using a fractional factorial design, the most significant growth factors influencing biomass production were found to be temperature, moisture content, inoculum concentration, and incubation period (p ≤ 0.05). Analysis of variance of a Box-Behnken design showed that the regression model was highly significant (p ≤ 0.05) with F-values of 10.38 (P = 0.0027, T. asperellum AU131) and 12.01 (p < 0.0017, T. longibrachiatum AU158). Under optimal conditions, maximum conidia yield of log10 (8.6) (T. asperellum AU131) and log10(9.18) (T. longibrachiatum) were obtained. For wettable powder Trichoderma species formulations, it was possible to maintain conidial viability at room temperature (25 °C) for eight months at concentrations above 106 CFU/g.

In-vitro compatibility assay of indigenous Trichoderma and Pseudomonas species and their antagonistic activities against black root rot disease (Fusarium solani) of faba bean (Vicia faba L.).

BACKGROUND: Faba bean (Vicia faba L.) cultivation is highly challenged by faba bean black root rot disease (Fusarium solani) in high lands of Ethiopia. To ensure sustainable production of faba beans, searching for eco-friendly disease management options is necessary to curb the progress of the disease timely. The indigenous biocontrol agents that suit local environments may effectively strive with in-situ microorganisms and suppress local pathogen strains. This study aimed to screen antagonistic indigenous compatible Trichoderma and Pseudomonas strains against Fusarium solani. In the pathogenicity test, soil-filled pots were arranged in complete random block design and sown with health faba bean seeds. The effect of some fungicides was evaluated against Fusarium by food poisoning methods to compare with the biocontrol agents. The antagonistic efficacy of biocontrol agents and their compatibility was investigated on Potato dextrose agar medium. RESULTS: Fusarium solani AAUF51 strain caused an intense root rotting in faba bean plant. The effect of Mancozeb 80% WP at 300 ppm was comparable with Trichoderma and Pseudomonas strains against Fusarium. The mycelial growth of test the pathogen was significantly (P ≤ 0.05) reduced to 86.67 and 85.19% by Trichoderma harzianum AAUW1 and Trichoderma viridae AAUC22 strains in dual culture, respectively. The volatile metabolites of Pseudomonas aeruginosa AAUS31 (77.78%) found the most efficient in reducing mycelial growth of Fusarium followed by Pseudomonas fluorescens AAUPF62 (71.11%) strains. The cell-free culture filtrates of Pseudomonas fluorescens AAUPF62 and Pseudomonas aeruginosa AAUS31 were more efficient than the Trichoderma strain in reducing the growth of Fusarium isolates. There was no zone of inhibition recorded between Trichoderma harzianum AAUW1, Trichoderma viridae AAUC22, Pseudomonas aeruginosa AAUS31, and Pseudomonas fluorescens AAUPF62 strains, hence they were mutually compatible. CONCLUSIONS: The compatible Trichoderma and Pseudomonas strains showed antagonistic potentiality that could be explored for faba bean protection against black root rot disease and might have a future dual application as biocontrol agents.

Genetic and phenotypic parameter estimates for reproduction traits in indigenous Arsi-Bale goats.

The study was conducted to evaluate reproductive performances and estimate genetic parameters for reproduction traits in Arsi-Bale goats. A total of 792 kidding records collected from 2001 to 2007 were used. Parity of dam, year, season and type of kidding were investigated as fixed effects by PROC GLM of SAS. Derivative-Free Restricted Maximum Likelihood (DFREML) method was used to estimate genetic parameters by fitting four animal models. Parity of dam and year of kidding influenced (P < 0.05) all the traits. The overall means for age at first kidding (AFK), kidding interval (KI), litter size at birth (LSB), litter size at weaning (LSW), litter weight at birth (LWB), litter weight at weaning (LWW), abortion and dystocia were 574.9 ± 8.3 days, 280.0 ± 13.7 days, 1.6 ± 0.03, 1.37 ± 0.03, 3.7 ± 0.08 kg, 9.11 ± 0.38 kg, 3.8% and 0.13%, respectively. The estimates of direct additive heritability for the traits, except for abortion and dystocia, under the best model (direct animal for AFK and repeatability model for other traits) were 0.245 ± 0.19, 0.060 ± 0.08, 0.074 ± 0.05, 0.006 ± 0.05, 0.125 ± 0.05, 0.053 ± 0.07, respectively, while the corresponding permanent environmental effects were 0.00 ± 0.00, 0.07 ± 0.07, 0.08 ± 0.05, 0.172 ± 0.06, 0.03 ± 0.04 and 0.07 ± 0.05, respectively. Repeatability estimates for KI, LSB, LSW, LWB and LWW were 0.13, 0.15, 0.18, 0.16 and 0.12, respectively. Genetic correlations between reproductive traits vary from medium to high. Arsi-Bale goats have good reproductive performance with low incidence of reproductive disorder. Except for AFK, other traits have low estimates of heritabilities with high genetic correlation among the traits. Repeated measures of the traits are needed before deciding to keep or cull the animal.