Halotolerant Citrobacter sp. remediates salinity stress and promotes the growth of Vigna radiata (L) by secreting extracellular polymeric substances (EPS) and biofilm formation: a novel active cell for microbial desalination cell (MDC) / Citrobacter sp. halotolerante. remedia el estrés salino y promueve el crecimiento de Vigna radiata (L) mediante la secreción de sustancias poliméricas extracelulares (EPS) y la formación de biopelículas: una nueva célula activa para la célula microbiana de desalinización (MDC)

To address soil salinization and its impact on crop production, microbial desalination cells (MDCs) offer a promising solution. These bioelectrochemical systems integrate desalination and wastewater treatment through microbial activity. A halotolerant beneficial bacterial strain called Citrobacter sp. strain KUT (CKUT) was isolated from India’s salt desert Run of Kutch, Gujrat, highlighting its potential application in combating soil salinization. CKUT exhibits high salt tolerance and has the ability to produce extracellular polymeric substances (EPS) at a concentration of 0.04 mg/ml. It forms biofilm that enable it to withstand up to 10% NaCl concentration. Additionally, CKUT shows promise in remediating salinity levels, reducing it from 4.5 to 2.7 gL−1. These characteristics are driven by biofilm formation and EPS production. In an experiment where V. radiata L. seedlings were inoculated with CKUT, the treated plants exhibited enhanced chlorophyll content, growth, and overall plant characteristics compared to seedlings treated with sodium chloride (NaCl). These improvements included increased shoot length (150 mm), root length (40 mm), and biomass. This indicates that CKUT treatment has the potential to enhance the suitability of V. radiata and other crops for cultivation in saline lands, effectively addressing the issue of soil salinization. Furthermore, integrating CKUT into microbial desalination cells (MDCs) offers an opportunity for freshwater production from seawater, contributing to sustainable agriculture by promoting improved crop growth and increased yield in areas prone to salinity.(AU)

Halotolerant Citrobacter sp. remediates salinity stress and promotes the growth of Vigna radiata (L) by secreting extracellular polymeric substances (EPS) and biofilm formation: a novel active cell for microbial desalination cell (MDC).

To address soil salinization and its impact on crop production, microbial desalination cells (MDCs) offer a promising solution. These bioelectrochemical systems integrate desalination and wastewater treatment through microbial activity. A halotolerant beneficial bacterial strain called Citrobacter sp. strain KUT (CKUT) was isolated from India's salt desert Run of Kutch, Gujrat, highlighting its potential application in combating soil salinization. CKUT exhibits high salt tolerance and has the ability to produce extracellular polymeric substances (EPS) at a concentration of 0.04 mg/ml. It forms biofilm that enable it to withstand up to 10% NaCl concentration. Additionally, CKUT shows promise in remediating salinity levels, reducing it from 4.5 to 2.7 gL-1. These characteristics are driven by biofilm formation and EPS production. In an experiment where V. radiata L. seedlings were inoculated with CKUT, the treated plants exhibited enhanced chlorophyll content, growth, and overall plant characteristics compared to seedlings treated with sodium chloride (NaCl). These improvements included increased shoot length (150 mm), root length (40 mm), and biomass. This indicates that CKUT treatment has the potential to enhance the suitability of V. radiata and other crops for cultivation in saline lands, effectively addressing the issue of soil salinization. Furthermore, integrating CKUT into microbial desalination cells (MDCs) offers an opportunity for freshwater production from seawater, contributing to sustainable agriculture by promoting improved crop growth and increased yield in areas prone to salinity. HIGHLIGHTS : • Soil salinization reduces crop yield, including Vigna radiata L. • Citrobacter sp. strain KUT (CKUT) is a halotolerant bacterium isolated from the salt desert Run of Kutch, Gujarat, which can tolerate high salt concentrations. • CKUT mitigates salinity by producing extracellular polymeric substances (EPS) and forming biofilms. • CKUT treatment demonstrated increased plant growth, biomass, and chlorophyll content under salinity stress, showcasing its potential in microbial desalination cell (MDC) for enhancing crop yield in salinized soils.

A novel probiotic strain of Lactobacillus fermentum TIU19 isolated from Haria beer showing both in vitro antibacterial and antibiofilm properties upon two multi resistant uro-pathogen strains.

Probiotics with antimicrobial activity are gaining interest as a topic in the research field. Urinary tract infections (UTIs), acquired in the hospital or the community, are among the most prevalent infections. The emergence of multidrug resistance (MDR) uro-pathogens has made the current situation more critical in terms of global public health. To face this situation, in this study, Lactobacillus fermentum TIU19 (L. fermentum TIU19) was isolated and characterized as a new probiotic strain of the rice-based fermented beverage Haria. Subsequently, we also investigated its application as a biological agent that inhibits the growth of multidrug-resistant uro-pathogens, Escherichia coli, and Enterococcus faecalis. The results showed that, the isolated strain L. fermentum TIU19 was sensitive to all antibiotics tested except vancomycin and was devoid of virulence factors, such as haemolytic and gelatinase activities. Therefore, it may be considered safe for public health. It has many probiotic properties, such as survival in simulated gastrointestinal fluid, antioxidant activity, ß-galactosidase producing ability, high cell surface hydrophobicity, adhesion ability to epithelial cells, and strong biofilm producer. The growth inhibitory and antibiofilm activities were shown against two uro-pathogens. All these results suggest that L. fermentum TIU19 can be explored as a potential probiotic with antagonistic activity against MDR uro-pathogenic E. coli and E. faecalis.

Oxidative stress, DNA, and membranes targets as modes of antibacterial and antibiofilm activity of facile synthesized biocompatible keratin-copper nanoparticles against multidrug resistant uro-pathogens.

Escherichia coli and Enterococcus faecalis are two of the most prevalent uro-pathogens and are difficult to treat as they acquire multidrug-resistant traits. In this study, the main objective was to develop biocompatible copper nanoparticles using chicken feather keratin protein (CuNPs-K) and to investigate their impact on multidrug-resistant (MDR) uro-pathogens, E. coli and E. faecalis, under both single and mixed culture conditions. CuNPs-K were characterised by UV-Vis spectroscopy, dynamic light scattering, X-ray diffraction, Fourier transform infrared spectroscopy, and docking experiments. The MIC values of CuNPs-K against single and mixed planktonic cultures were 50 µg/ml and 75 µg/ml, respectively. CuNPs-K efficiently disrupted the biofilm of single and mixed uro-pathogen cultures by eliminating sessile cells. This biofilm disruption may be attributed to a decline in the production of extracellular polymeric substances in both single and mixed bacterial cultures treated with CuNPs-K. Moreover, selective antimicrobial activity was determined by selectivity assays using T24 cells. CuNPs-K targets both the bacterial membrane and DNA with elevated reactive oxygen species (ROS) as their bactericidal mode of action. This comprehensive antimicrobial activity of CuNPs-K was further confirmed in vivo by using the zebra fish model. In this study, CuNPs-K effectively reduced bacterial load with increased survivability of infected zebrafish. All these results suggest that CuNPs-K can be explored as an exceptional antibacterial agent against MDR uro-pathogenic E. coli and E. faecalis.

Genotoxic impact of emerging contaminant amoxicillin residue on zebra fish (Danio rerio) embryos.

The widest-spectrum, most-consumed ß-lactam antibiotic amoxicillin (AMX) is used to treat bovine mastitis that is caused primarily by many bacteria. Excessive use of antibiotics can lead to established residual contamination of the milk even after pasteurization. The amount of antibiotic residue above Maximum Residue Limit (MRL) has a negative impact on both public health and the environment. Therefore, the objective of this study is to determine the concentration of amoxicillin residue (AMXR) in raw and pasteurized milk samples of cow suffered from mastitis, by the standard methods of HPLC compared to pure AMX drug and effect of the said residue on the developmental toxicity and genotoxicity of zebra fish at 72 hpf and 48 hpf embryo, respectively. Results obtained by HPLC showed that AMXR exhibits 574.89 and 250.75 times higher concentration in the raw and pasteurized milk than MRL in compare to pure AMX drug. This current study showed that AMXR decreased the body length and yolk sac region, while the pure AMX drug-treated group showed increased height and length of the yolk sac and shorter body length relative to the other groups. The Comet Assay measured the DNA damage caused by AMXR. The group where AMXR were applied showed the highest percentage of tail DNA and tail moment relative to other groups. So, here AMXR is considered as the genotoxic contaminant that is emerging and affect on public health.

Allelochemical catechol comprehensively impedes bacterial blight of rice caused by Xanthomonas oryzae pv. oryzae.

Xanthomonas oryzae pv. oryzae (Xoo) induces bacterial leaf blight (BLB), is known to be the most devastating disease of rice. The present investigation for the first time explains the antibacterial, anti-biofilm, and antivirulence potential of the simplest allelochemical catechol. Bacterial viability and growth are significantly reducing in catechol treatment. Further study also reveals that catechol also inhibits primary attachment and preformed biofilm of Xoo even at half MIC concentration. The half MIC concentration of catechol also induce a significant decrease in virulence factors like swimming, swarming, exopolysaccharide, and xanthomonadin production. Next, we investigate the possible antibacterial mode of action of catechol against Xoo. Results show that, the catechol caused oxidative stress and targets cell membrane for its antibacterial activity. Whereas, in silico study reveals that, catechol binds with the catalytic domain of XanA protein and this may be consider as a reason for antibiofilm activity of catechol. Moreover, in virulence assay on rice plants, we observe significant decrement in lesion length in catechol and Xoo co-treated rice leaves as compared with only Xoo treated leaves. All the results clearly show, allelochemical catechol to be a potential compound for the antibacterial, anti-biofilm, and antivirulence agent against Xoo and consequently mitigating the BLB disease advancement in rice.

Photodynamic antimicrobial chemotherapy (PACT) using riboflavin inhibits the mono and dual species biofilm produced by antibiotic resistant Staphylococcus aureus and Escherichia coli.

PURPOSE: Multispecies biofilms play a significant role in persistent infections. Furthermore, by interspecies transfer of antibiotic resistance genes, multispecies biofilms spread antibiotic resistance. The purpose of this study was to investigate the effect of Photodynamic Antimicrobial Chemotherapy (PACT) using riboflavin on mono and multi species biofilms. METHODS: For this we used two clinically relevant opportunistic pathogens species E. coli and S. aureus as mono-species and multispecies biofilms. We did broth dilution assay for antibacterial, crystal violet assay for biofilms and fluorometric study for reactive oxygen species (ROS) and extracellular polymeric substance (EPS) production by phenol-HCl method. RESULTS: Antibacterial study revealed that photo-illuminated riboflavin shows bactericidal effect against each bacteria and their mix culture. E. coli was found to be little more resistant than S. aureus. Crystal violet assay revealed photo-illuminated riboflavin shows anti-biofilms activity against both mono and mix species biofilms. But mix species biofilms were more resistant to PACT than mono species biofilms. Further study revealed this may be due to the interaction between different EPS production, hence in mix species biofilms EPS production is less affected after PACT than mono species biofilms. We found photo-illuminated riboflavin increased the intracellular ROS production. CONCLUSION: Photo-illuminated riboflavin shows bactericidal and anti-biofilms effect against each bacteria and their mix culture. Photo-illuminated increased intracellular ROS production, which may induce the oxidative stress and destroy the respiratory system of bacteria.

Sodium alginate-based edible coating containing nanoemulsion of Citrus sinensis essential oil eradicates planktonic and sessile cells of food-borne pathogens and increased quality attributes of tomatoes.

There is a growing interest from the worldwide scientific community in formulating edible- biodegradable coatings to replace non-biodegradable and expensive commercial wax-based coatings for preserving postharvest quality attributes of vegetables including tomatoes. Postharvest tomatoes are a suspected vehicle for both Salmonella and Listeria in food poisoning incidents. In this work, the effectiveness of edible nano-emulsion coatings containing sweet orange essential oil and sodium alginate were prepared and characterized, then evaluated antibacterial and antibiofilm activity against Salmonella and Listeria and simultaneously, examined its coating effect on various quality characteristics of tomatoes at 22 ± 2 °C over a 15 days storage period. DLS (Dynamic light scattering) study revealed stable nanoemulsion formulation with 43.23 nm particle size. The high whiteness index of nanoemulsion has a positive impact on product marketability and desirability. Antibacterial and antibiofilm studies revealed nanoemulsion effectively eradicate both sessile and planktonic forms of Salmonella and Listeria in both single and multi-species culture conditions. Tomatoes coated with edible coating significantly enhanced firmness up to 33%, decreased total mesophilic bacteria including Salmonella and Listeria, and reduced weight loss up to 3 fold lower than uncoated one. Sensory analysis revealed that the use of the edible coating increased the total acceptance scores of tomatoes.

Antibacterial, anti-biofilm activity and mechanism of action of pancreatin doped zinc oxide nanoparticles against methicillin resistant Staphylococcus aureus.

Staphylococcus aureus are known to cause diseases from normal skin wound to life intimidating infections. Among the drug resistant strain, management of methicillin resistant Staphylococcus aureus (MRSA) is very difficult by using conventional antibiotic treatment. Both Zinc oxide nanoparticles (ZnONPs) and pancreatin (PK) are known to have antibacterial activity. Our main objective is to dope PK on ZnONPs to reduced zinc-oxide toxicity but increased anti-bacterial and anti-biofilms activity. In present study, we showed that, functions of zinc oxide nanoparticles with pancreatin enzyme (ZnONPs-PK) have anti-bacterial, anti-biofilms, anti-motility and anti-virulence properties against MRSA. Moreover, ZnONPs-PK were more potent to eradicate MRSA than only ZnONPs and PK. Application of the produced nano-composites as treatment on infected swine dermis predominantly reflects the potential treatment property of it. The vancomycin sensitivity of MRSA was significantly increased on application with ZnONPs-PK. Further study revealed cell membrane was the target of the ZnONPs-PK and that leads to oxidative damage of the cells. The produced nanoparticles were found completely non-toxic to human's keratinocytes and lung epithelial cell lines at its bactericidal concentration. Overall, this study emphasizes the potential mechanisms underlying the selective bactericidal properties of ZnONPs-PK against MRSA. This novel nanoparticle strategy may provide the ideal solution for comprehensive management of MRSA and its associated diseases with minimising the use of antibiotics.

Water as carrier of information of heat shock and drug effect between two groups of Adhatoda vasica plants / Água como veículo de informação sobre choque térmico e efeito medicamentoso em dois grupos de plantas Adhatoda vasica / Agua como vehículo de información sobre shock térmico y efecto medicamentoso en dos grupos de plantas Adhatoda vasica

Adhatoda vasica Nees plants were grown in 50 earthen pots, which were divided into 5 batches A, B, C, D, and E. Of these A, B and C, D were arranged into two separate parallel pairs. One leaf of each plant of an adjacent pair was immersed in sterile tap water in a beaker. Adjacent beakers in each pair A B or C D were connected by polythene tubes containing wet cotton threads. One leaf of each plant of A was given heat shock by immersing a leaf in hot water for 5 min. One leaf of each plant of C was treated with Cantharis vesicatoria 200c. Batch E served as the unstressed and untreated control. One hour after heat shock or drug treatment all the leaves were harvested and their proteins were extracted by chilled protein extraction buffer. Proteins were separated by Fast Protein Liquid Chromatography (FPLC). Protein profiles of A, B and C, D showed marked similarity with respect to expression and repression of some proteins. It is concluded that the effect of heat shock and drug treatment is transmitted through water in the capillaries of cotton threads connecting the pairs of plants. It is assumed that heat shock or drug treatment altered locally the water structure in the leaves which was propagated through global network of water structure over the protein network in the whole plants, and from there to the interfacial water in the beakers and cotton threads. A homeopathic potency is thought to be specifically structured water which influences the water structure in the treated organism(AU)

Plantas Adhatoda vasica Nees foram cultivadas em 50 vasos de argila e divididos em 5 lotes A, B, C, D e E. Os lotes A, B e C, D foram arranjados em dois pares paralelos separados. Uma folha de cada planta dos pares adjacentes foi submergida num béquer com água corrente estéril. Os béqueres de cada par, A, B e C, D foram interconectados através de tubos de polietileno contendo fio de algodão umedecido. Uma folha de cada planta no lote A recebeu choque térmico através de imersão em água quente por 5 minutos. Uma folha de cada planta no lote C foi tratada com Cantharis vesicatoria 200c. O lote E foi utilizado como controle não submetido nem a estresse nem a medicação. Uma hora depois da aplicação de choque térmico ou tratamento medicamentoso, todas as folhas foram coletadas e suas proteínas foram extraídas através de tampão de extração de proteínas frio. As proteínas foram separadas através de cromatografia líquida rápida de proteínas (CLRP). Os perfis proteicos dos lotes A, B, C e D mostraram similaridade considerável quanto à expressão e repressão de algumas proteínas. Conclui-se que o efeito do choque térmico e do tratamento medicamentoso foi transmitido através da água nos capilares de fio de algodão conectando pares de plantas. Assume-se que o choque térmico e o tratamento medicamentoso alteraram localmente a estrutura da água nas folhas, que se propagou através da rede global da estrutura da água até a rede proteica nas plantas inteiras e dali para a água interfacial nos béqueres e fios de algodão. Considera-se que uma potência homeopática é água especificamente estruturada que influencia a estrutura da água no organismo tratado.(AU)

Water as carrier of information of heat shock and drug effect between two groups of Adhatoda vasica plants / Água como veículo de informação sobre choque térmico e efeito medicamentoso em dois grupos de plantas Adhatoda vasica / Agua como vehículo de información sobre shock térmico y efecto medicamentoso en dos grupos de plantas Adhatoda vasica

Adhatoda vasica Nees plants were grown in 50 earthen pots, which were divided into 5 batches A, B, C, D, and E. Of these A, B and C, D were arranged into two separate parallel pairs. One leaf of each plant of an adjacent pair was immersed in sterile tap water in a beaker. Adjacent beakers in each pair A B or C D were connected by polythene tubes containing wet cotton threads. One leaf of each plant of A was given heat shock by immersing a leaf in hot water for 5 min. One leaf of each plant of C was treated with Cantharis vesicatoria 200c. Batch E served as the unstressed and untreated control. One hour after heat shock or drug treatment all the leaves were harvested and their proteins were extracted by chilled protein extraction buffer. Proteins were separated by Fast Protein Liquid Chromatography (FPLC). Protein profiles of A, B and C, D showed marked similarity with respect to expression and repression of some proteins. It is concluded that the effect of heat shock and drug treatment is transmitted through water in the capillaries of cotton threads connecting the pairs of plants. It is assumed that heat shock or drug treatment altered locally the water structure in the leaves which was propagated through global network of water structure over the protein network in the whole plants, and from there to the interfacial water in the beakers and cotton threads. A homeopathic potency is thought to be specifically structured water which influences the water structure in the treated organism

Plantas Adhatoda vasica Nees foram cultivadas em 50 vasos de argila e divididos em 5 lotes A, B, C, D e E. Os lotes A, B e C, D foram arranjados em dois pares paralelos separados. Uma folha de cada planta dos pares adjacentes foi submergida num béquer com água corrente estéril. Os béqueres de cada par, A, B e C, D foram interconectados através de tubos de polietileno contendo fio de algodão umedecido. Uma folha de cada planta no lote A recebeu choque térmico através de imersão em água quente por 5 minutos. Uma folha de cada planta no lote C foi tratada com Cantharis vesicatoria 200c. O lote E foi utilizado como controle não submetido nem a estresse nem a medicação. Uma hora depois da aplicação de choque térmico ou tratamento medicamentoso, todas as folhas foram coletadas e suas proteínas foram extraídas através de tampão de extração de proteínas frio. As proteínas foram separadas através de cromatografia líquida rápida de proteínas (CLRP). Os perfis proteicos dos lotes A, B, C e D mostraram similaridade considerável quanto à expressão e repressão de algumas proteínas. Conclui-se que o efeito do choque térmico e do tratamento medicamentoso foi transmitido através da água nos capilares de fio de algodão conectando pares de plantas. Assume-se que o choque térmico e o tratamento medicamentoso alteraram localmente a estrutura da água nas folhas, que se propagou através da rede global da estrutura da água até a rede proteica nas plantas inteiras e dali para a água interfacial nos béqueres e fios de algodão. Considera-se que uma potência homeopática é água especificamente estruturada que influencia a estrutura da água no organismo tratado.

Sepia 200cH at 1: 1000 dilution ameliorates salt stress in cowpea seedlings but its medium 90% ethanol proves ineffective at the same dilution

Soil salinity severely affects crop yield all over the world. In a recent study we observed that Natrum mur 200cH, a homeopathic remedy, improved growth in germinating cowpea seeds. In the present study we have tested another remedy Sepia, which is complementary to Natrum mur, on cowpea seedlings under salt stress. Cowpea seedlings grown over moist filter paper in petridishes were divided into 4 groups: (1) control in sterile water, (2) in 50mM NaCl solution, (3) seeds pretreated with 90% ethanol diluted with water 1:100 and then transferred to 50mM NaCl solution, (4) seeds pretreated with Sepia 200cH diluted with water 1:100 and transferred to 50mM NaCl solution. In another experiment the groups were same, but the dilution of 90% ethanol and Sepia 200cH was 1:1000 instead of 1:100. The purpose was to further reduce the ethanol content in both the drug and its vehicle 90% ethanol, so that the alcohol effect is minimized or abolished. The data were analysed by ANOVA followed by t-test. Sepia 200cH at both 1:100 and 1:1000 dilutions significantly increased growth, sugar, chlorophyll, protein and water content in seedlings as compared to the untreated salt-stressed group. The effect with the1000th dilution of Sepia 200cH was more pronounced than with its 100th dilution. The vehicle 90% ethanol at 1:100 dilution produced some positive effect on the seedlings, but the 1000th dilution of the vehicle produced no such effect. It is, therefore, concluded that Sepia 200cH could ameliorate salt stress in cowpea seedlings and that the 1000th dilution is more effective than its 100th dilution. The alcohol effect is totally eliminated with the 1000th dilution of 90% ethanol. Thus the 1000th dilution could retain the drug effect and eliminate the vehicle effect. (AU)

Introdução: A salinidade do solo afeta seriamente a produtividade das culturas no mundo todo. Num estudo recente, observamos que o medicamento homeopático Natrum muriaticum 200 Ch melhorou o crescimento de sementes de feijão-fradinho Vigna unguiculata (L.) Walp] em germinação. Objetivo: Nesse trabalho, testamos o medicamento homeopático Sepia succus, complementar de Nat-m, em plântulas de feijão-fradinho submetidas a estresse salino. Métodos: Plântulas de feijão-fradinho desenvolvidas sobre papel de filtro umedecido em placas de Petri foram divididas em 4 grupos: (1) controle em água estéril, (2) em solução de NaCl 50 mM, (3) sementes pré-tratadas com etanol a 90% diluído 1:100 em água e após transferido a solução de NaCl 50mM, e (4) sementes pré-tratadas com Sep 200 cH diluídas 1:100 em água e após transferidas a solução de NaCl 50 mM. Num segundo experimento, os grupos foram os mesmos, mas a diluição do etanol a 90% e de Sep 200 cH foi 1:1.000 ao invés de 1:00 para reduzir ainda mais o teor de etanol em ambos, medicamento e veículo a fim de minimizar ou abolir o efeito do álcool. Os dados foram analisados através de ANOVA seguida de teste t de Student. Resultados: As duas diluições de Sep 200 cH, 1:100 e 1:1.000, aumentaram significativamente o crescimento, teor de açúcar, clorofila, proteína e água das plântulas por comparação ao grupo submetido a estresse salino não tratado. O efeito da 1.000ª diluição de Sep 200 cH foi mais pronunciado que o da 100ª diluição. O veículo etanol a 90% em diluição 1:100 induziu alguns efeitos positivos nas plântulas, mas a diluição 1:1.000 não apresentou esses efeitos. Conclusões: Sep 200 cH reverteu o efeito de estresse salino nas plântulas de feijão-fradinho, sendo que a 1.000ª diluição foi mais efetiva que a 100ª. O efeito do álcool foi totalmente eliminado com a 1.000ª diluição do etanol a 90%. Portanto, a 1.000ª conservou o efeito do medicamento e eliminou o efeito do veículo. (AU)

Sepia 200cH at 1: 1000 dilution ameliorates salt stress in cowpea seedlings but its medium 90 por cento ethanol proves ineffective at the same dilution

Soil salinity severely affects crop yield all over the world. In a recent study we observed that Natrum mur 200cH, a homeopathic remedy, improved growth in germinating cowpea seeds. In the present study we have tested another remedy Sepia, which is complementary to Natrum mur, on cowpea seedlings under salt stress. Cowpea seedlings grown over moist filter paper in petridishes were divided into 4 groups: (1) control in sterile water, (2) in 50mM NaCl solution, (3) seeds pretreated with 90% ethanol diluted with water 1:100 and then transferred to 50mM NaCl solution, (4) seeds pretreated with Sepia 200cH diluted with water 1:100 and transferred to 50mM NaCl solution. In another experiment the groups were same, but the dilution of 90% ethanol and Sepia 200cH was 1:1000 instead of 1:100. The purpose was to further reduce the ethanol content in both the drug and its vehicle 90% ethanol, so that the alcohol effect is minimized or abolished. The data were analysed by ANOVA followed by t-test. Sepia 200cH at both 1:100 and 1:1000 dilutions significantly increased growth, sugar, chlorophyll, protein and water content in seedlings as compared to the untreated salt-stressed group. The effect with the1000th dilution of Sepia 200cH was more pronounced than with its 100th dilution. The vehicle 90% ethanol at 1:100 dilution produced some positive effect on the seedlings, but the 1000th dilution of the vehicle produced no such effect. It is, therefore, concluded that Sepia 200cH could ameliorate salt stress in cowpea seedlings and that the 1000th dilution is more effective than its 100th dilution. The alcohol effect is totally eliminated with the 1000th dilution of 90% ethanol. Thus the 1000th dilution could retain the drug effect and eliminate the vehicle effect.

Introdução: A salinidade do solo afeta seriamente a produtividade das culturas no mundo todo. Num estudo recente, observamos que o medicamento homeopático Natrum muriaticum 200 Ch melhorou o crescimento de sementes de feijão-fradinho Vigna unguiculata (L.) Walp] em germinação. Objetivo: Nesse trabalho, testamos o medicamento homeopático Sepia succus, complementar de Nat-m, em plântulas de feijão-fradinho submetidas a estresse salino. Métodos: Plântulas de feijão-fradinho desenvolvidas sobre papel de filtro umedecido em placas de Petri foram divididas em 4 grupos: (1) controle em água estéril, (2) em solução de NaCl 50 mM, (3) sementes pré-tratadas com etanol a 90% diluído 1:100 em água e após transferido a solução de NaCl 50mM, e (4) sementes pré-tratadas com Sep 200 cH diluídas 1:100 em água e após transferidas a solução de NaCl 50 mM. Num segundo experimento, os grupos foram os mesmos, mas a diluição do etanol a 90% e de Sep 200 cH foi 1:1.000 ao invés de 1:00 para reduzir ainda mais o teor de etanol em ambos, medicamento e veículo a fim de minimizar ou abolir o efeito do álcool. Os dados foram analisados através de ANOVA seguida de teste t de Student. Resultados: As duas diluições de Sep 200 cH, 1:100 e 1:1.000, aumentaram significativamente o crescimento, teor de açúcar, clorofila, proteína e água das plântulas por comparação ao grupo submetido a estresse salino não tratado. O efeito da 1.000ª diluição de Sep 200 cH foi mais pronunciado que o da 100ª diluição. O veículo etanol a 90% em diluição 1:100 induziu alguns efeitos positivos nas plântulas, mas a diluição 1:1.000 não apresentou esses efeitos. Conclusões: Sep 200 cH reverteu o efeito de estresse salino nas plântulas de feijão-fradinho, sendo que a 1.000ª diluição foi mais efetiva que a 100ª. O efeito do álcool foi totalmente eliminado com a 1.000ª diluição do etanol a 90%. Portanto, a 1.000ª conservou o efeito do medicamento e eliminou o efeito do veículo.