Serological and epidemiological investigation of Mycobacterium avium subspecies paratuberculosis in bovines in Pakistan.

Objective: To investigate the prevalence of paratuberculosis in cattle and buffaloes at twelve public dairy farms in Punjab, Pakistan. Methods: A total of 2181 more than two-year-old animals (1242 cattle and 939 buffaloes) were tested by avian tuberculin, i.e., killed purified protein derivative (PPD) of Mycobacterium avium paratuberculosis and indirect ELISA. Blood and fecal samples were collected from tuberculin positive animals. These samples were further processed by indirect ELISA. The data were analyzed using frequency analysis and logistic analysis procedures. Results: The prevalence of paratuberculosis at public dairy farms was 3.8%, as determined by tuberculin + ELISA test. It varied from 0.71-13.5% with a 100% herd prevalence. Multivariate logistic regression analysis revealed that species, milk production, total animals, total small ruminants, and total buffaloes were significantly associated with the occurrence of paratuberculosis. Odd ratio analysis revealed that with a one-kilogram increase in body weight, there will be a 0.006% increase in disease occurrence. With the increase in one animal in small ruminants and buffaloes, there will be 0.008% and 0.42% greater chances of developing paratuberculosis, respectively. Bivariate logistic regression analysis of cattle and buffaloes revealed that farm number, age, and total number of cattle were significantly associated with the occurrence of paratuberculosis. A one-month increase in lactation length increases the chance of tuberculosis by 0.004%; similarly, a one-liter increase in milk production increases the chance of disease by 10%. With each additional buffalo in the herd, there will be a 0.007% greater chance for the occurrence of paratuberculosis. Conclusion: This study concluded that tuberculin testing can be used in conjunction with ELISA to screen animals for paratuberculosis in countries with scarce resources, such as Pakistan. The efficacy of disease diagnosis can be improved by combining multiple tests.

In vitro silicon supplementation enhanced acclimatisation and growth of sugarcane (Saccharum officinarum) via improved antioxidant and nutrient acquisition patterns in saline soil.

Salinity affects crop growth by modulating cellular ionic concentrations and generation of reactive oxygen species. Application of silicon (Si) has proved beneficial in ameliorating salinity-triggered plant growth and yield retardations. Leaf roll explants of three sugarcane (Saccharum officinarum ) genotypes (HSF-240, CPF-246, CPF-250) were cultured in Murashige and Skoog (MS) medium supplemented with K2 SiO3 . In vitro regenerated plantlets were acclimatised and grown in natural saline soil. In absence of Si, cv. CPF-246 exhibited better salt tolerance as indicted by maximum chlorophyll a and chlorophyll b contents, rate of photosynthesis and root K+ uptake along with less cellular hydrogen peroxide content. Silicon restricted root Na+ uptake but assisted in K+ , Ca2+ , Mg2+ and Fe2+ accretion in roots and their translocation towards shoots. Cv. HSF-240 and cv. CPF-250 exhibited more increase in photosynthetic pigment content, stomatal conductance and photosynthetic rate after addition of 25 or 50mgL-1 Si than control group. Optimum phenolic content and antioxidant enzyme activity along with decreased lipid peroxidation and hydrogen peroxide content were recorded in all three sugarcane genotypes raised in presence of 25 or 50mgL-1 Si. These findings signify Si supplementation (50mgL-1 ) in tissue culture medium and plant adaptation in saline soil. Further in vitro studies involving Si-mediated gene expression modulations in sugarcane protoplasts shall assist in deciphering cross-talk between Si uptake and cellular responses. The application of Si can further be tested for other plant species to devise strategies for improved crop growth and utilisation of saline areas for crop cultivation.

Molecular cloning, biological description, and functional analysis of Ajfos transcription factor in pathogen-induced Apostichopus japonicus.

Activator protein-1 subfamily member c-Fos wields significant influence over cellular activities, such as regulation of cell growth and division, cell death, and immune responses under various extracellular situations. In this study, the full-length c-Fos of sea cucumber, Apostichopus japonicus (Ajfos) was successfully cloned and analyzed. The anticipated 306 amino acid sequences of Ajfos displayed a basic-leucine zipper (bZIP) domain, similar to invertebrate counterparts. In addition, the qPCR results suggested Ajfos expressed in all tissues, with the highest level in coelomocytes from polian vesicle (vesicle lumen cells), followed by coelomocytes from coelom (coelomocytes). Moreover, the expression levels of Ajfos in the coelomocytes and vesicle lumen cells of sea cucumber showed significant changes after the Vibrio splendidus challenge, especially reaching a peak at 6 h. Compared with the silencing negative control RNA interference (siNC) group, silencing Ajfos (siAjfos) in vivo decreased the downstream proliferation-related gene expression of vesicle lumen cells after infection with V. splendidus while no significant influence was observed on coelomocytes. Furthermore, the proliferation proportion of vesicle lumen cells in the siAjfos group was significantly reduced under pathogen stimulation conditions. Finally, based on the fluctuation trend of total coelomocyte density (TCD) from coelom and polian vesicle previously discovered, it is evident that Ajfos played a critical role in facilitating the swift proliferation of vesicle lumen cells in response to V. splendidus stimulation. Altogether, this research provided an initial reference of the function of Ajfos in echinoderms, unveiling its participation in host coelomocyte proliferation of sea cucumbers during bacterial challenges.

A Participatory Approach in Assessing the Knowledge, Attitude, and Practices (KAP) of Stakeholders and Livestock Owners about Ticks and Tick-Borne Diseases from Sindh, Pakistan.

Ticks and tick-borne diseases (TTBDs) are responsible for significant losses in terms of treatment costs, decreased productivity (such as milk and meat), reduced reproductive ability, and financial crisis to livestock owners. In Pakistan, it is crucial to periodically assess the risk of TTBDs and ecological factors, potential causes of acaricidal resistance in tick fauna, and the intensive increase in the spread of TTBDs. Participatory epidemiological approaches are key to assessing the livestock owners' and stakeholders' knowledge, attitude, and practices (KAP) about TTBDs. The current study determined the KAP about ticks and tick-borne diseases of respondents from Sindh, Pakistan. A total of 240 respondents were interviewed from different ecological zones: among them, 42.5% (n = 102) of the respondents practiced the manual removal of ticks from animals, while acaricide usage was indicated by 137 respondents (57.0%) as occurring sometimes, 50 (20.8%) monthly, 41 (17.0%) fortnightly, and 12 (5%) weekly, during the peak infestation season. Ticks were 2.6 times [OR = 2.5 (95% Cl = 1.47-4.06)] and viruses were 1.89 times [OR = 188 (95% Cl = 1.09-2.9)] more likely to cause the development of disease in animals than any other pathogen. Despite the appropriate usage of acaricides, the knowledge of participants was inadequate. The findings of this study emphasize the need to take into account identified knowledge gaps and to take the initiative in carrying out appropriate education activities and extension programs to enhance the adoption of effective tick prevention and control strategies.

Transcriptome analysis reveals key transcription factors and pathways of polian vesicle associated with cell proliferation in Vibrio splendidus-challenged Apostichopus japonicus.

Polian vesicle is thought to produce coelomocytes and contribute to the sea cucumber's immune system. Our previous work has indicated that polian vesicle was responsible for cell proliferation at 72 h post pathogenic challenge. However, the transcription factors related to the activation of effector factors and the molecular process behind this remained unknown. In this study, to reveal the early functions of polian vesicle in response to the microbe, a comparative transcriptome sequencing of polian vesicle in V. splendidus-challenged Apostichopus japonicus, including normal group (PV 0 h), pathogen challenging for 6 h (PV 6 h) and 12 h (PV 12 h) was performed. Compared PV 0 h to PV 6 h, PV 0 h to PV 12 h, and PV 6 h to PV 12 h, we found 69, 211, and 175 differentially expressed genes (DEGs), respectively. KEGG enrichment analysis revealed the DEGs, including several transcription factors such as fos, FOS-FOX, ATF2, egr1, KLF2, and Notch3 between PV 6 h and PV 12 h were consistently enriched in MAPK, Apelin and Notch3 signaling pathways related to cell proliferation compared with that in PV 0 h. Important DEGs involved in cell growth were chosen, and their expression patterns were almost the same as the transcriptome profile analysis by qPCR. Protein interaction network analysis indicated that two DEGs of fos and egr1 were probably significant as key candidate genes controlling cell proliferation and differentiation in polian vesicle after pathogenic infection in A. japonicus. Overall, our analysis demonstrates that polian vesicles may play an essential role in regulating proliferation via transcription factors-mediated signaling pathway in A. japonicus and provide new insights into hematopoietic modulation of polian vesicles in response to pathogen infection.

Health risk assessment and bioaccumulation of potentially toxic metals from water, soil, and forages near coal mines of district Chakwal, Punjab, Pakistan.

Water, forages, and soil contamination with potentially toxic metals (PTMs) through anthropogenic activities has become a significant environmental concern. It is crucial to find out the level of PTMs in water, soil, and forages near industrial areas. The PTMs enter the body of living organisms through these sources and have become a potential risk for humans and animals. Therefore, the present study aims at the health risk assessment of PTMs and their accumulation in soil, water, and forages of three tehsils (Kallar Kahar, Choa Saidan Shah, and Chakwal) in district Chakwal. Samples of wastewater, soil, and forages were collected from various sites of district Chakwal. PTMs detected in the present study were cadmium (Cd), chromium (Cr), lead (Pb), zinc (Zn), cobalt (Co), copper (Cu), and nickel (Ni), and their levels were measured through atomic absorption spectrophotometer (AAs GF95 graphite furnace auto sampler). Pollution load index (PLI), bio concentration factor (BCF), soil enrichment factors (EF), daily intake value (DIM), and health risk index (HRI) in sheep, cow, and buffalo were also analyzed. The results revealed that the mean concentration (mg/L) of Cd (0.72-0.91 mg/L), Cr (1.84-2.23 mg/L), Pb (0.95-3.22 mg/L), Co (0.74-2.93 mg/L), Cu (0.84-1.96 mg/L), and Ni (1.39-4.39 mg/L) in wastewater samples was higher than permissible limits set by WHO, NEQS, WWF, USEPA, and Pakistan in all three tehsils of district Chakwal. Similarly, in soil samples, concentrations of Cd (1.21-1.95 mg/kg), Cr (38.1-56.4 mg/kg), and Ni (28.3-55.9 mg/kg) were higher than their respective threshold values. The mean concentration of PTMs in forage samples (Parthenium hysterophorus, Mentha spicata, Justicia adhatoda, Calotropis procera, Xanthium strumarium, Amaranthaceae sp.) showed that maximum values of Cd (5.35-7.55 mg/kg), Cr (5.47-7.51 mg/kg), Pb (30-36 mg/kg), and Ni (12.6-57.5 mg/kg) were beyond their safe limit set for forages. PLI, BCF, and EF were > 1.0 for almost all the PTMs. The DIM and HRI for sheep were less than < 1.0 but for cows and buffalo were > 1.0. The current study showed that soil, water, and forages near coal mines area are contaminated with PTMs which enter the food chain and pose significant harm to humans and animals. In order to prevent their dangerous concentration in the food chain, regular assessment of PTMs present in soil, forages, irrigating water, and food is recommended.

Biodegradation of monocrotophos by Brucella intermedia Msd2 isolated from cotton plant.

Widespread and inadequate use of Monocrotophos has led to several environmental issues. Biodegradation is an ecofriendly method used for detoxification of toxic monocrotophos. In the present study, Msd2 bacterial strain was isolated from the cotton plant growing in contaminated sites of Sahiwal, Pakistan. Msd2 is capable of utilizing the monocrotophos (MCP) organophosphate pesticide as its sole carbon source for growth. Msd2 was identified as Brucella intermedia on the basis of morphology, biochemical characterization and 16S rRNA sequencing. B. intermedia showed tolerance of MCP up to 100 ppm. The presence of opd candidate gene for pesticide degradation, gives credence to B. intermedia as an effective bacterium to degrade MCP. Screening of the B. intermedia strain Msd2 for plant growth promoting activities revealed its ability to produce ammonia, exopolysaccharides, catalase, amylase and ACC-deaminase, and phosphorus, zinc and potassium solubilization. The optimization of the growth parameters (temperatures, shaking rpm, and pH level) of the MCP-degrading isolate was carried out in minimal salt broth supplemented with MCP. The optimal pH, temperature, and rpm for Msd2 growth were observed as pH 6, 35 °C, and 120 rpm, respectively. Based on optimization results, batch degradation experiment was performed. Biodegradation of MCP by B. intermedia was monitored using HPLC and recorded 78% degradation of MCP at 100 ppm concentration within 7 days of incubation. Degradation of MCP by Msd2 followed the first order reaction kinetics. Plant growth promoting and multi-stress tolerance ability of Msd2 was confirmed by molecular analysis. It is concluded that Brucella intermedia strain Msd2 could be beneficial as potential biological agent for an effective bioremediation for polluted environments.

Elucidating molecular characterization of chlorpyrifos and profenofos degrading distinct bacterial strains for enhancing seed germination potential of Gossypium arboreum L.

Chlorpyrifos (CP) and profenofos (PF) are organophosphate pesticides (OPs) widely used in agriculture and are noxious to both fauna and flora. The presented work was designed to attenuate the toxicity of both pesticides in the growth parameters of a cotton crop by applying plant growth-promoting rhizobacteria (PGPR), namely Pseudomonas aeruginosa PM36 and Bacillus sp. PM37. The multifarious biological activities of both strains include plant growth-promoting traits, including phosphate solubilization; indole-3-acetic acid (IAA), siderophore, and HCN production; nitrogen fixation; and enzymatic activity such as cellulase, protease, amylase, and catalase. Furthermore, the molecular profiling of multi-stress-responsive genes, including acdS, ituC, czcD, nifH, and sfp, also confirmed the plant growth regulation and abiotic stress tolerance potential of PM36 and PM37. Both strains (PM36 and PM37) revealed 92% and 89% of CP degradation at 50 ppm and 87% and 81% at 150 ppm within 7 days. Simultaneously 94% and 98% PF degradation was observed at 50 ppm and 90% and 92% at 150 ppm within 7 days at 35 °C and pH 7. Biodegradation was analyzed using HPLC and FTIR. The strains exhibited first-order reaction kinetics, indicating their reliance on CP and PF as energy and carbon sources. The presence of opd, mpd, and opdA genes in both strains also supported the CP and PF degradation potential of both strains. Inoculation of strains under normal and OP stress conditions resulted in a significant increase in seed germination, plant biomass, and chlorophyll contents of the cotton seedling. Our findings indicate that the strains PM36 and PM37 have abilities as biodegraders and plant growth promoters, with potential applications in crop sciences and bioremediation studies. These strains could serve as an environmentally friendly, sustainable, and socially acceptable solution to manage OP-contaminated sites.

Microwave-assisted sustainable exploration of cocklebur leaves (Xanthium strumarium L.) as a novel source of distinct yellow natural colorant for dyeing cotton fabric.

During current times, the use of bio-colorants attained public acceptance as a sustainable alternative to synthetic ones which in turn reduced the environmental contamination. The present study focused on the green, safe, and clean technology for the resurgence of natural colorant from cocklebur (Xanthium strumarium L.) leaves and their application to cotton fabric. Natural colorants were extracted by employing an eco-friendly microwave-assisted extraction process using an aqueous and alkaline medium. Dyeing of cotton fabric was carried out using irradiated and unirradiated cotton fabric with irradiated and unirradiated natural dyes of cocklebur leaves. The results of extraction experiments revealed that 4 min microwave-assisted alkaline extract exhibited significantly outstanding color strength onto microwave-treated cotton fabric compared to aqueous one. Further to investigate the optimum dyeing conditions for cotton fabric, various dyeing variables such as dyeing time, dyeing temperature, dye concentration, and exhausting agent were monitored and found a superior result using a dye concentration of 45 ml, for dyeing cotton fabric at 75 °C for 50 min in the presence of 4 g/100 ml of table salt. For improvement in color strength and color fastness properties, the effects of various bio-mordants, such as eucalyptus bark, acacia bark, turmeric rhizome, and onion shells, and chemical mordants (aluminum and copper) on dyed cotton fabric were also evaluated. It was also observed that cotton fabric dyed with alkaline extract of cocklebur leaves using bio-mordants as pre-mordants (4% acacia, 4% eucalyptus, 2% onion) and post-mordants (3% onion, 3% eucalyptus, 4% acacia) exhibited the highest color strength and various hues with acceptable colorfastness properties against light, washing, and rubbing in comparison to chemical mordants. The ISO standard for fastness also revealed that bio-mordanting has enhanced the rating from good to excellent in comparison to chemical mordants. The results provide ample scope for the extraction of yellow natural dye from the cocklebur leaves for eco-friendly coloration of fabrics using bio-mordants.

Acinetobacter schindleri SR-5-1 decipher morpho-physio-biochemical and nutritional improvements to Pisum sativum L. and Linum usitatissimum L. maintained under wastewater/cadmium stress.

Metal retention in wastewater fertigated crops poses a potential hazard to food chain. Current work demonstrates the bioremediation and growth-promoting potential of Acinetobacter schindleri SR-5-1 by using nitrogen-fixing (pea) and non-nitrogen fixing (linseed) plants under cadmium (Cd) and wastewater irrigation regimes. Both plants were grown at 250 or 500 CdCl2 and 75 or 100% wastewater, each separately with and without A. schindleri SR-5-1 inoculation. The results revealed that Cd and wastewater significantly decreased growth, biomass, antioxidants, and nutrient acquisition through increased malondialdehyde, H2O2, and Cd accumulation. However, application of A. schindleri SR-5-1 significantly promoted morpho-physio-biochemical attributes while diminishing MDA and H2O2 under applied Cd and wastewater stress levels in both pea and linseed. Further, PGPR inoculation positively influenced pea and linseed seedlings through a substantial decline in Cd accumulation in roots/shoots and retained the optimal level of essential nutrients. It was inferred that both pea and linseed, with A. schindleri SR-5-1 application, exhibited higher growth and metabolism under Cd and wastewater stress but substantial tolerance was acquired under wastewater stress. Studied plants exhibited tolerance in order of 75% WW ≥ 250 µM Cd ≥ 100%WW ≥ 500 µM Cd treatment under A. schindleri inoculation. Current findings revealed the potential of A. schindleri to be exploited both for bioremediation and bio-fertilization under Cd, and wastewater-polluted regimes to reduce metal contamination of edible plants. It was suggested that with inoculation of A. schindleri SR-5-1, 75% WW dilution can be applied for irrigation of both nitrogen-fixing and non-nitrogen-fixing crops.

Application of zinc oxide nanoparticles immobilizes the chromium uptake in rice plants by regulating the physiological, biochemical and cellular attributes.

Zinc oxide nano particles (ZnO NPs) have been employed as a novel strategy to regulate plant tolerance and alleviate heavy metal stress, but our scanty knowledge regarding the systematic role of ZnO NPs to ameliorate chromium (Cr) stress especially in rice necessitates an in-depth investigation. An experiment was performed to evaluate the effect of different concentrations of ZnO NPs (e.g., 0, 25, 50, 100 mg/L) in ameliorating the Cr toxicity and accumulation in rice seedlings in hydroponic system. Our results demonstrated that Cr (100 µM) severely inhibited the rice seedling growth, whereas exogenous treatment of ZnO NPs significantly alleviated Cr toxicity stress and promoted the plant growth. Moreover, application of ZnO NPs significantly augmented the germination energy, germination percentage, germination index, and vigor index. In addition, biomass accumulation, antioxidants (SOD, CAT, POD), nutrient acquisition (Zn, Fe) was also improved in ZnO NPs-treated plants, while the lipid peroxidation (MDA, H2O2), electrolyte leakage as well as Cr uptake and in-planta accumulation was significantly decreased. The burgeoning effects were more apparent at ZnO NPs (100 mg/L) suggesting the optimum treatment to ameliorate Cr induced oxidative stress in rice plants. Furthermore, the treatment of ZnO NPs (100 mg/L) reduced the level of endogenous abscisic acid (ABA) and stimulated the growth regulator hormones such as brassinosteroids (BRs) possibly linked with enhanced phytochelatins (PCs) levels. The ultrastructure analysis at cellular level of rice revealed that the application of 100 mg/L ZnO NPs protected the chloroplast integrity and other cell organells via improvement in plant ionomics, antioxidant activities and down regulating Cr induced oxidative stress in rice plants. Conclusively, observations of the current study will be helpful in developing stratigies to decrease Cr contamination in food chain by employing ZnO NPs and to mitigate the drastic effects of Cr in plants for the sustainable crop growth.

Microbial and enzymatic battle with food contaminant zearalenone (ZEN).

Zearalenone (ZEN) contamination of various foods and feeds is an important global problem. In some animals and humans, ZEN causes significant health issues in addition to massive economic losses, annually. Therefore, removal or degradation of the ZEN in foods and feeds is required to be done. The conventional physical and chemical methods have some serious issues including poor efficiency, decrease in nutritional value, palatability of feed, and use of costly equipment. Research examined microbes from diverse media for their ability to degrade zearalenone and other toxins, and the findings of several investigations revealed that enzymes produced from microbes play a significant role in the degradation of mycotoxins. In established bacterial hosts, genetically engineered technique was used to enhance heterologously produced degrading enzymes. Then, the bio-degradation of ZEN by the use of micro-organisms or their enzymes is much more advantageous and is close to nature and ecofriendly. Furthermore, an effort is made to put forward the work done by different scientists on the biodegradation of ZEN by the use of fungi, yeast, bacteria, and/or their enzymes to degrade the ZEN to non-toxic products. KEY POINTS: •Evolved microbial strains degraded ZEA more quickly •Different degrading properties were studied.

Ameliorative Effects of Anti-Clostridial Egg Yolk Antibodies (IgYs) in Experimentally-Induced Avian Necrotic Enteritis.

The present study was planned to evaluate the ameliorative effects of egg yolk antibodies (EYAs) in broiler chicken. For this purpose, 80-day-old broiler chickens were divided into four groups (A-D), where group A was kept as negative control. Experimental infection with C. perfringens (1 × 108 cfu/mL) was induced via oral route on days 17, 18 and 19 of the experiment in groups B, C and D. Groups C and D were passively immunized by anti-clostridial IgYs @ 1 mL per bird via oral and oral and intramuscular (I/M) routes respectively, on days 21 to 24, and on days 22 and 24 of the experiment, respectively. Two necropsies were performed (the first on day 26th and the second on day 35th). Birds in group B showed behavioral signs e.g., laziness, depression and diarrhea, gross post-mortem lesions e.g., increase in the relative weights (RW), due to acute swelling and congestion of liver and kidneys and ballooning and hemorrhages of jejunum and microscopic lesions e.g., congestion and necrosis in liver and kidneys' parenchyma and disrupted epithelium with fewer goblet cells in jejunum, compared to the group A. Birds in groups C and D, showed significant improvements in clinical and behavioral signs, RW of liver, kidneys and jejunum, swelling, congestion and mononuclear cells' infiltration in liver and kidneys and damages in the jejunal-wall, compared to group B. The most significant changes were found in birds of group C. Our study revealed ameliorative effects of EYAs on certain biological parameters however, further studies would be needed to justify a safer production and a reliable application of EYAs in NE outbreaks.

Silicate Inhibits the Cytosolic Influx of Chloride in Protoplasts of Wheat and Affects the Chloride Transporters, TaCLC1 and TaNPF2.4/2.5.

Chloride is an essential nutrient for plants, but high concentrations can be harmful. Silicon ameliorates both abiotic and biotic stresses in plants, but it is unknown if it can prevent cellular increase of chloride. Therefore, we investigated the influx of Cl- ions in two wheat cultivars different in salt sensitivity, by epifluorescence microscopy and a highly Cl--sensitive dye, MQAE, N-[ethoxycarbonylmethyl]-6-methoxy-quinolinium bromide, in absence and presence of potassium silicate, K2SiO3. The Cl--influx was higher in the salt-sensitive cv. Vinjett, than in the salt-tolerant cv. S-24, and silicate pre-treatment of protoplasts inhibited the Cl--influx in both cultivars, but more in the sensitive cv. Vinjett. To investigate if the Cl--transporters TaCLC1 and TaNPF2.4/2.5 are affected by silicate, expression analyses by RT-qPCR were undertaken of TaCLC1 and TaNPF 2.4/2.5 transcripts in the absence and presence of 100 mM NaCl, with and without the presence of K2SiO3. The results show that both transporter genes were expressed in roots and shoots of wheat seedlings, but their expressions were differently affected by silicate. The TaNPF2.4/2.5 expression in leaves was markedly depressed by silicate. These findings demonstrate that less chloride accumulates in the cytosol of leaf mesophyll by Si treatment and increases salt tolerance.

Deciphering distinct root exudation, ionomics, and physio-biochemical attributes of Serratia marcescens CP-13 inoculated differentially Cd tolerant Zea mays cultivars.

Cadmium (Cd) being a non-essential, mobile, and toxic heavy metal, negatively affects the plant growth and physiology. Current work investigated the impact of Serratia marcescens CP-13 inoculation on root organic acids and nutrient exudates of two maize cultivars varying in Cd tolerance under induced Cd toxicity. Seedlings of Cd-sensitive (Sahiwal-2002) and Cd-tolerant (MMRI-Yellow) cultivars were grown either inoculated or non-inoculated with CP-13 in Petri plates having various Cd stress levels (0, 6, 12, 18, 24, 30 µM). Seedlings were transferred to rhizoboxes for the collection of root exudates and analysis of physio-biochemical traits. Both maize cultivars exuded higher organic acids and nutrient exudates under non-inoculated conditions as compared to inoculated ones. Non-inoculated tolerant cultivar exhibited higher nutrient accumulation, biomass, antioxidants, total chlorophyll, Cd release meanwhile reduced Cd uptake, lipid peroxidation, exudation of organic acids, and nutrients than the sensitive one. However, under CP-13 inoculation, Cd sensitive cultivar exhibited less exudation of organic acids (citric acid, acetic acid, malic acid, glutamic acid, formic acid, succinic acid, and oxalic acid), nutrients mobilization (K, Na, Zn, Ca, and Mg), total chlorophyll, antioxidants (APX, SOD, POD), total soluble sugar, diminished MDA, and Cd uptake. The significant reduction in release of root exudates by both cultivars was likely due to the plant growth promoting traits of CP-13 which confer Cd tolerance. The maximum release of rhizospheric root exudates were documented at 30 µM applied Cd stress. Therefore, the Serratia sp. CP-13 was proposed as a potential inoculant for bioremediation of Cd together with maize cultivars.

Deciphering distinct biological control and growth promoting potential of multi-stress tolerant Bacillus subtilis PM32 for potato stem canker.

Plant growth-promoting rhizobacteria (PGPR) represent a set of microorganisms that play significant role in improving plant growth and controlling the phytopathogens. Unpredictable performance after the application of PGPR has been observed when these were shifted from in-vitro to in-vivo conditions due to the prevalence of various abiotic stress conditions. During growing period, the potato crop is subjected to a combination of biotic and abiotic stresses. Rhizoctonia solani, a soil-borne plant pathogen, causes reduced vigor and yield of potato crop worldwide. In the current study, multi-stress-tolerant rhizobacterial strain, Bacillus subtilis PM32, was isolated from field-grown potato with various plant growth promoting (PGP) traits including zinc and potassium solubilization, biological nitrogen fixation, ammonia and siderophore, as well as extracellular enzyme productions (cellulase, catalase, amylase, protease, pectinase, and chitinase). The strain PM32 exhibited a distinct potential to support plant growth by demonstrating production of indole-3-acetic acid (102.6 µM/mL), ACC-deaminase activity (1.63 µM of α-ketobutyrate/h/mg protein), and exopolysaccharides (2.27 mg/mL). By retarding mycelial growth of R. solani the strain PM32 drastically reduced pathogenicity of R. solani. The strain PM32 also suppressed the pathogenic activity significantly by impeding mycelial expansion of R. solani with inhibition co-efficient of 49.87. The B. subtilis PM32 also depicted significant tolerance towards salt, heavy metal (Pb), heat and drought stress. PCR based amplification of ituC and acds genes coding for iturin and ACC-deaminase activity respectively indicated potential of strain PM32 for lipopeptides production and ACC deaminase enzyme activity. Results of both in-vitro and pot experiments under greenhouse conditions depicted the efficiency of B. subtilis PM32 as a promising bio-control agent for R. solani infection together with enhanced growth of potato plants as deciphered from biomass accumulation, chlorophyll a, b, and carotenoid contents. Therefore, it was envisioned that application of indigenous multi-stress tolerant PGPR may serve to induce biotic and abiotic stress tolerance in crops/plants for pathogen control and sustainable global food supply. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01067-2.

Long-term copper exposure promotes apoptosis and autophagy by inducing oxidative stress in pig testis.

Copper (Cu) is a heavy metal which is being used widely in the industry and agriculture. However, the overuse of Cu makes it a common environmental pollutant. In order to investigate the testicular toxicity of Cu, the pigs were divided into three groups and were given Cu at 10 (control), 125, and 250 mg/kg body weight, respectively. The feeding period was 80 days. Serum hormone results showed that Cu exposure decreased the concentrations of follicular stimulating hormone (FSH) and luteinizing hormone (LH) and increased the concentration of thyroxine (T4). Meanwhile, Cu exposure upregulated the expression of Cu transporter mRNA (Slc31a1, ATP7A, and ATP7B) in the testis, leading to increase in testicular Cu and led to spermatogenesis disorder. The Cu exposure led to an increased expression of antioxidant-related mRNA (Gpx4, TRX, HO-1, SOD1, SOD2, SOD3, CAT), along with increase in the MDA concentration in the testis. In LG group, the ROS in the testis was significantly increased. Furthermore, the apoptotic-related mRNA (Caspase3, Caspase8, Caspase9, Bax, Cytc, Bak1, APAF1, p53) and protein (Active Caspase3) and the autophagy-related mRNA (Beclin1, ATG5, LC3, and LC3B) expression increased after Cu exposure. The mitochondrial membrane potential in the testicular tissue decreased, while the number of apoptotic cells increased, as a result of oxidative stress. Overall, our study indicated that the Cu exposure promotes testicular apoptosis and autophagy by mediating oxidative stress, which is considered as the key mechanism causing testicular degeneration as well as dysfunction.

Influence of foliar glutathione and putrescine on metabolism and mineral status of genetically diverse rapeseed cultivars under hexavalent chromium stress.

We studied the physio-biochemical involvement of exogenous signaling compounds, glutathione and putrescine (alone and in combination), on three contrasting genotypes (cvs. Shiralee, Rainbow, and Dunkled) of canola (Brassica napus L.) of plants exposed to chromium stress. Seeds were germinated in Cr-contaminated soil (0 and 50 µg/g Cr6+), and both signaling compounds were applied as a foliar spray to 20-day-old plants. Changes in root, stem, and leaf nitro-oxidative metabolism, endogenous GSH level, secondary metabolites, and mineral nutrients were investigated from 60-day-old plants. Exposure to Cr6+ increased stem GSH and NO concentrations in all cultivars. Maximum root Cr6+ bioaccumulation was recorded in cv. Rainbow and the least in cv. Shiralee. Also, Cr6+ stress decreased number and weight of seeds and pod length. Disturbances in root and shoot mineral profile were evident; however, its magnitude varied in all cultivars. The exogenous GSH improved root and shoot P, Fe, S, and Zn concentrations; however, the effect was cultivar specific. Leaf endogenous GSH was increased by exogenous GSH while NO levels remained unaffected. The GSH application also promoted shoot Cr6+ bioaccumulation while PUT application caused a recovery in seed number and seed weight. Both PUT and GSH differentially affected tissue-specific secondary metabolite profile. Overall, the exogenous GSH was much more effective in alleviating the Cr+6 toxicity in canola.

Elucidating Cd-mediated distinct rhizospheric and in planta ionomic and physio-biochemical responses of two contrasting Zea mays L. cultivars.

Cadmium (Cd) in soil-plant system can abridge plant growth by initiating alterations in root zones. Hydroponics and rhizoboxes are useful techniques to monitor plant responses against various natural and/or induced metal stresses. However, soil based studies are considered more appropriate in order to devise efficient food safety and remediation strategies. The present research evaluated the Cd-mediated variations in elemental dynamics of rhizospheric soil together with in planta ionomics and morpho-physio-biochemical traits of two differentially Cd responsive maize cultivars. Cd-sensitive (31P41) and Cd-tolerant (3062) cultivars were grown in pots filled with 0, 20, 40, 60 and 80 µg/kg CdCl2 supplemented soil. The results depicted that the maize cultivars significantly influenced the elemental dynamics of rhizosphere as well as in planta mineral accumulation under applied Cd stress. The uptake and translocation of N, P, K, Ca, Mg, Zn and Fe from rhizosphere and root cell sap was significantly higher in Cd stressed cv. 3062 as compared to cv. 31P41. In sensitive cultivar (31P41), Cd toxicity resulted in significantly prominent reduction of biomass, leaf area, chlorophyll, carotenoids, protein contents as well as catalase activity in comparison to tolerant one (3062). Analysis of tolerance indexes (TIs) validated that cv. 3062 exhibited advantageous growth and efficient Cd tolerance due to elevated proline, phenolics and activity of antioxidative machinery as compared to cv. 31P41. The cv. 3062 exhibited 54% and 37% less Cd bio-concentration (BCF) and translocation factors (TF), respectively in comparison to cv. 31P41 under highest Cd stress regime. Lower BCF and TF designated a higher Cd stabilization by tolerant cultivar (3062) in rhizospheric zone and its potential use in future remediation plans.