Plant growth-promoting bacteria isolated from earthworms enhance spinach growth and its phytoremediation potential in metal-contaminated soils / Las bacterias promotoras del crecimiento vegetal aisladas de lombrices de tierra mejoran el crecimiento de las espinacas y su potencial de fitorremediación en suelos contaminados con metales

The aim of this study was to evaluate the impact of metal-tolerant plant growth-promoting bacteria (PGPB) isolated from the chloragogenous tissue of Aporrectodea molleri, which represents a unique habitat. Our objectives were to investigate their effects on the growth of Spinacia oleracea under heavy metal stress and assess their potential for enhancing phytoremediation capabilities. The experiment was conducted in an alkaline soil contaminated with 7 mg kg-1 of cadmium, 100 mg kg-1 of nickel, 150 mg kg-1 of copper, 300 mg kg-1 of Zinc, and mg kg-1 of 600 Manganese. The results showed that heavy metal stress considerably diminished root (42.8%) and shoot length (60.1%), biomass (80%), chlorophyll content (41%), soil alkaline (45%), and acid (51%) phosphatases (42%) and urease (42%). However, soil inoculation with bacterial isolates remarkably improved plant growth. Soil bioaugmentation increased spinach growth (up to 74.5% for root length, up to 106.3% for shoot length, and up to 5.5 folds for fresh biomass) while significantly increasing soil enzyme activity and NPK content. Multivariate data analysis indicated that soil inoculation with Bacillus circulans TC7 promoted plant growth while limiting metal bioaccumulation, whereas Pseudomonas sp. TC33 and Bacillus subtilis TC34 increased metal bioaccumulation in spinach tissues while minimizing their toxicity. Our study confirms that earthworms are a reservoir of multi-beneficial bacteria that can effectively improve phytoremediation efficiency and mitigate the toxic effects of heavy metals on plant growth. Further studies are needed to investigate the long-term effects and feasibility of using these isolates as a consortium in field applications.(AU)

Plant growth-promoting bacteria isolated from earthworms enhance spinach growth and its phytoremediation potential in metal-contaminated soils.

The aim of this study was to evaluate the impact of metal-tolerant plant growth-promoting bacteria (PGPB) isolated from the chloragogenous tissue of Aporrectodea molleri, which represents a unique habitat. Our objectives were to investigate their effects on the growth of Spinacia oleracea under heavy metal stress and assess their potential for enhancing phytoremediation capabilities. The experiment was conducted in an alkaline soil contaminated with 7 mg kg-1 of cadmium, 100 mg kg-1 of nickel, 150 mg kg-1 of copper, 300 mg kg-1 of Zinc, and mg kg-1 of 600 Manganese. The results showed that heavy metal stress considerably diminished root (42.8%) and shoot length (60.1%), biomass (80%), chlorophyll content (41%), soil alkaline (45%), and acid (51%) phosphatases (42%) and urease (42%). However, soil inoculation with bacterial isolates remarkably improved plant growth. Soil bioaugmentation increased spinach growth (up to 74.5% for root length, up to 106.3% for shoot length, and up to 5.5 folds for fresh biomass) while significantly increasing soil enzyme activity and NPK content. Multivariate data analysis indicated that soil inoculation with Bacillus circulans TC7 promoted plant growth while limiting metal bioaccumulation, whereas Pseudomonas sp. TC33 and Bacillus subtilis TC34 increased metal bioaccumulation in spinach tissues while minimizing their toxicity. Our study confirms that earthworms are a reservoir of multi-beneficial bacteria that can effectively improve phytoremediation efficiency and mitigate the toxic effects of heavy metals on plant growth. Further studies are needed to investigate the long-term effects and feasibility of using these isolates as a consortium in field applications.

Earthworm Aporrectodea molleri (oligochaeta)'s coelomic fluid-associated bacteria modify soil biochemical properties and improve maize (Zea mays L.) plant growth under abiotic stress conditions.

This study evaluated the impact of Aporrectodea molleri's coelomic fluid-associated bacteria (CFB) on Zea mays L. growth and soil biochemical characteristics under abiotic stress conditions, including alkaline soil (pH = 8) and nitrogen (N), phosphate (P), and potassium (K) deficit. Compared to maize cultivated in uninoculated soil, the effect of CFB on boosting plant growth under abiotic stress was notably exceptional. Different CFB treatments increased significantly root and shoot length by 50% and 21%, respectively. Furthermore, the presence of isolates in soil resulted in a significant increase in plant fresh and dry weights (of up to 113% and 91% for roots, and up to 173% and 44% for shoots), leaf surface (78%), and steam diameter (107%). Overall, soil inoculation with CFB significantly (P < 0.05) enhanced chlorophyll and water content in the plant compared to the untreated soil. Despite the soil's alkaline condition, CFB drastically boosted soil quality by increasing nutrient availability (up to 30 ppm for N, 2 ppm for P, and 60 ppm for K) and enzyme activity (up to 1.14 µg p-NP h-1 g-1 for acide phosphatase, 9 µg p-NP h-1 g-1 for alkaline phosphatase and 40 µg NH4-N 2 h-1 g-1 for urease), throughout the early stages of the growth period. Interestingly, alkaline phosphatase concentrations were substantially greater in treatments with different isolates than acid phosphatase. Furthermore, the principal component analysis showed that the inoculation with bacteria strains CFB1 Buttiauxella gaviniae and CFB3 Aeromonas hydrophila had a significantly better stimulatory stimulatory and direct influence on maize growth than the other isolates had a substantial effect on soil's biochemical features. Thus, we assumed that the beneficial contribution of earthworms in the rhizosphere might be attributed in large part to associated microorganisms.

Taxonomic and functional characteristics of aerobic bacteria isolated from the chloragogenous tissue of the earthworm Aporrectodea molleri.

Earthworms are considered as a rich microhabitat for the growth and proliferation of diverse soil microorganisms. Hence, earthworms' associated bacteria attracted interest due to their high metabolic profiles and benefits to soil fertility and plant growth. In this study, we aimed to isolate for the first-time aerobic bacteria present in the chloragogenous tissue of the earthworm Apporectodea molleri and test their Plant Growth-Promoting abilities and their resistance to heavy metals (Mn, Zn, Cu, Cd, and Ni). The 16S rRNA gene sequencing revealed the affiliation of the fifteen isolates to six main bacterial genera: Enterobacter, Citrobacter, Aeromonas, Pseudomonas, Bacillus, Terribacillus. These strains displayed different plant growth promoting traits (e.g., indole-3-acetic acid IAA, siderophores, nitrogen fixation, phosphate, and potassium solubilization), in addition, they were able to resist differently to heavy metals. Bacillus strains were most effective as three strains, namely B. subtilis strain TC34; B. circulans strain TC7 and Bacillus sp. strain TC10, were positive to all PGP traits and resisted to all heavy metals. This study illustrates the potential of bacteria from the chloragogenous tissue to exhibit multiple properties, which can be related to the functional feature of this tissue to stock metabolites and neutralize toxic elements.

Earthworms and their cutaneous excreta can modify the virulence and reproductive capability of entomopathogenic nematodes and fungi.

Earthworms are ecological engineers that can contribute to the displacement of biological control agents such as the entomopathogenic nematodes (EPNs) and fungi (EPF). However, a previous study showed that the presence of cutaneous excreta (CEx) and feeding behavior of the earthworm species Eisenia fetida (Haplotaxida: Lumbricidae) compromise the biocontrol efficacy of certain EPN species by reducing, for example, their reproductive capability. Whether this phenomenon is a general pattern for the interaction of earthworms-entomopathogens is still unknown. We hypothesized that diverse earthworm species might differentially affect EPN and EPF infectivity and reproductive capability. Here we investigated the interaction of different earthworm species (Eisenia fetida, Lumbricus terrestris, and Perionyx excavatus) (Haplotaxida) and EPN species (Steinernema feltiae, S. riojaense, and Heterorhabditis bacteriophora) (Rhabditida) or EPF species (Beauveria bassiana and Metarhizium anisopliae) (Hypocreales), in two independent experiments. First, we evaluated the application of each entomopathogen combined with earthworms or their CEx in autoclaved soil. Hereafter, we studied the impact of the earthworms' CEx on entomopathogens applied at two different concentrations in autoclaved sand. Overall, we found that the effect of earthworms on entomopathogens was species-specific. For example, E. fetida reduced the virulence of S. feltiae, resulted in neutral effects for S. riojaense, and increased H. bacteriophora virulence. However, the earthworm P. excavates increased the virulence of S. feltiae, reduced the activity of H. bacteriophora, at least at specific timings, while S. riojaense remained unaffected. Finally, none of the EPN species were affected by the presence of L. terrestris. Also, the exposure to earthworm CEx resulted in a positive, negative or neutral effect on the virulence and reproduction capability depending on the earthworm-EPN species interaction. Concerning EPF, the impact of earthworms was also differential among species. Thus, E. fetida was detrimental to M. anisopliae and B. bassiana after eight days post-exposure, whereas Lumbricus terrestris resulted only detrimental to B. bassiana. In addition, most of the CEx treatments of both earthworm species decreased B. bassiana virulence and growth. However, the EPF M. anisopliae was unaffected when exposed to L. terrestris CEx, while the exposure to E. fetida CEx produced contrasting results. We conclude that earthworms and their CEx can have positive, deleterious, or neutral impacts on entomopathogens that often coinhabit soils, and that we must consider the species specificity of these interactions for mutual uses in biological control programs. Additional studies are needed to verify these interactions under natural conditions.

Assessment and comparison of the pathogenicity of Sheeppox Virus strains isolated in Morocco.

BACKGROUND AND OBJECTIVES: Sheeppox virus causes systemic disease in sheep that is often associated with high morbidity and mortality. Protection against sheep pox is mainly based on medical prophylaxis, vaccination being the only way. In Morocco, and up to now, there is no available information about local challenge strain to use for controlling the efficiency of vaccines produced against sheep pox. Hence, the objective of the present study was to evaluate and compare the pathogenicity of seven Sheeppox virus (SPVs) isolates from 1993-1995 in Morocco. MATERIALS AND METHODS: These seven SPV isolates have undergone various tests to evaluate their pathogenicity: Passages and titration on cell culture, Experimental inoculation on sheep, Virus-neutralization, In vivo titration and viral re-isolation by real-time PCR assay. RESULTS: All infected lambs showed severe clinical signs, while most of them have been reproduced on 5 dpi and persisted until 21 dpi. The lambs infected by Oj1P4, Oj2P4 and BerP5 appeared lethargic, reluctant to move compared to those infected by other isolates. The results also revealed that all isolates were able to induce serological response. Virus isolation from infected organs and blood and amplification of the viral DNA by real-time PCR proved the presence of the virus in tissues and blood of infected lambs. These Moroccan SPVs demonstrated that the three isolates Oj1P4, Oj2P4 and BerP5 have a high pathogenicity; especially the BerP5 isolate which has an important infectious titer. CONCLUSION: These results demonstrate that the Berkane isolate is the most pathogenic of the tested isolates and it can be an excellent challenge strain for the control of the efficiency of vaccines against sheep pox produced in Morocco.