Levilactobacillus brevis MZ384011 and Levilactobacillus brevis MW362779 can mitigate lead induced hepato-renal damage by regulating visceral dispersion and fecal excretion.

Heavy metal pollution is a global issue. Current study provides evidence on Pb toxicity ameliorative potential and safe nature of Levilactobacillus brevis MZ384011 (S1) and Levilactobacillus brevis MW362779 (S2), isolated from carnivore gut and human milk, respectively. In a 60-days experiment, the rats were distributed into six groups. G-I, G-V and G-VI were kept on normal diet, while GII-IV were fed on lead nitrate (500 mg/kg) supplemented food, throughout experiment. After confirmation of Pb toxicity in GII-IV at 15th day, S1 was orally administered to G-III and G-V while S2 was given to G-IV and G-VI at a dose of 1 × 109 CFU/animal/day. On day 60 of experiment, positive control (G-II) displayed significant reduction in body weight, total protein, albumin, globulin, mineral profile, erythrocyte count, hemoglobin, hematocrit and hematological indices and elevation in leukocyte count, alanine aminotransferase, aspartate aminotransferase, bilirubin, uric acid and creatinine along with alterations in hepato-renal architecture. With reference to G-II, the G-III and G-IV displayed significant improvement in all aforementioned parameters, 40-60% reduction in tissue Pb levels (blood, liver, kidney and adipose tissue) and elevation in fecal Pb contents (p = 0.000). The groups V and VI did not show any sign of toxicity. The findings confirm that strains are safe for biological application and can reverse Pb toxicity by facilitating fecal Pb excretion and reducing its systemic dispersal. To best of our information this is the first report on Pb toxicity ameliorative role of Levilactobacillus brevis from human milk, the safest source.

Biogenic Synthesis, Characterization and Antibacterial Properties of Silver Nanoparticles against Human Pathogens.

Biogenic synthesis of silver nanoparticles (AgNPs) is more eco-friendly and cost-effective approach as compared to the conventional chemical synthesis. Biologically synthesized AgNPs have been proved as therapeutically effective and valuable compounds. In this study, the four bacterial strains Escherichia coli (MT448673), Pseudomonas aeruginosa (MN900691), Bacillus subtilis (MN900684) and Bacillus licheniformis (MN900686) were used for the biogenic synthesis of AgNPs. Agar well diffusion assay revealed to determine the antibacterial activity of all biogenically synthesized AGNPs showed that P. aeruginosa AgNPs possessed significantly high (p < 0.05) antibacterial potential against all tested isolates. The one-way ANOVA test showed that that P. aeruginosa AgNPs showed significantly (p < 0.05) larger zones of inhibition (ZOI: 19 to 22 mm) compared to the positive control (rifampicin: 50 µg/mL) while no ZOI was observed against negative control (Dimethyl sulfoxide: DMSO). Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) concentration against four test strains also showed that among all biogenically synthesized NPs, P. aeruginosa AgNPs showed effective MIC (3.3-3.6 µg/mL) and MBC (4.3-4.6 µg/mL). Hence, P. aeruginosa AGNPs were characterized using visual UV vis-spectroscopy, X-ray diffractometer (XRD), fourier transform infrared (FTIR) and scanning electron microscopy (SEM). The formation of peak around 430 nm indicated the formation of AgNPs while the FTIR confirmed the involvement of biological molecules in the formation of nanoparticles (NPs). SEM revealed that the NPs were of approximately 40 nm. Overall, this study suggested that the biogenically synthesized nanoparticles could be utilized as effective antimicrobial agents for effective disease control.

Linking traditional anti-ulcer use of rhizomes of Bergenia ciliata (Haw.) to its anti-Helicobacter pylori constituents.

The rhizomes of Bergenia ciliata (B. ciliata, Family: Saxifragaceae) are widely used for treating gastric ulcers in folk medicine in Asia. It was hypothesized that anti-ulcer activity of B. ciliata is due to its anti-Helicobacter pylori (H. pylori) activity. The anti-H. pylori activity was investigated on six clinical bacterial isolates using agar well-diffusion and broth micro-dilution methods. The anti-H. pylori activity of amoxicillin (standard) was the highest (Zone of inhibition; ZI = 25 mm, minimum inhibitory concentration; MIC=0.125 µg/µL) whereas among all the extracts of the rhizomes, methanol extract showed the highest activity (ZI = 16 mm, MIC = 12.50 µg/µL). Bioassay guided isolation of methanol extract using chromatographic and crystallization techniques isolated bergenin (ZI = 21mm, MIC = 0.391µg/µL) as constituent responsible for anti-H. pylori activity. The present study describes for the first time anti-H. pylori activity and possible mechanism of anti-ulcer properties of rhizomes of B. ciliata.

Antilisterial efficacy of Lactobacillus brevis MF179529 from cow: an in vivo evidence.

BACKGROUND: Listeria monocytogenes is an opportunistic foodborne pathogen that causes human Listeriosis and high mortality particularly in immunocompromised individuals. Pregnant women are more prone to L. monocytogenes infection resulting in abortions. In the present study, antilisterial activity of Lactobacillus brevis (LB) MF179529, a probiotic bacterial strain, was investigated in a murine model. METHODS: Initially a pilot study was conducted to determine the dose of L. monocytogenes required to cause symptomatic listeriosis. In the main trial, mice were divided into 4 groups. Group I was kept as negative control, group II was exposed to L. monocytogenes and maintained as positive control. Group III was fed with L. brevis only, while group IV received L. brevis for 3 days prior to L. monocytogenes infection. A volume of 200 µl of L. monocytogenes ATCC 19115 and L. brevis MF179529 bacterial suspension corresponding to cell density of 109CFU/ml were given to respective groups by intragastric route. Progress of infection was monitored for 7 days including general health scoring, listeria dispersion in organs, bacterial load in intestine and blood biochemistry were recorded on 3rd, 5th and 7th days post infection (dpi). RESULTS: Clinical listeriosis was induced by 109CFU/ml of L. monocytogenes ATCC 19115 in mice. Animals of group IV displayed minor signs of infection. L. brevis supplementation resulted in significant reduction in dispersion and propagation of L. monocytogenes in liver, spleen and intestine. L. brevis MF179529 consumption led to a significant elevation of number of lactic acid bacteria and reduction of total plate count, anaerobic count and coliform population in intestine. Moreover, total leukocyte and neutrophil counts of treated animals were similar to the negative control while positive control group displayed higher number. Safety evaluation of L. brevis was performed by monitoring general health, hematological and serological parameters of L. brevis fed and negative control group (group III and I). No significant difference in feed intake, body temperature, body weight and blood picture could be detected in L. brevis supplemented and control groups. CONCLUSION: Our results indicate ameliorative role of L. brevis in L. monocytogenes infection and suggest that L. brevis could be used for prophylactic measure.

Toxicological Evaluation of Essential Oils from Some Plants of Rutaceae Family.

Essential oils are produced as secondary metabolites by aromatic plants, predominantly belonging to families Apiaceae, Lamiaceae, Myrtaceae, and Rutaceae. The family Rutaceae has great economic importance for its numerous edible fruits and essential oils. In the present study, essential oils of seven plants of family Rutaceae, Aegle marmelos, Murraya koenigii, Citrus reticulata Blanco, Zanthoxylum armatum, Skimmia laureola, Murraya paniculata, and Boenninghausenia albiflora, were used for their toxicological assessment. Seven groups of selected essential oils-treated Wistar rats were established against control group (n = 5) that received water for 14 days; animals were offered feed and water ad libitum and treated with essential oils at 400 mg/kg body weight. Hematological studies revealed significant elevation in TEC in animals treated with essential oils of M. koenigii, S. laureola, and B. albiflora, while an elevation in PCV and depletion in MCV were observed in animals treated with M. paniculata and B. albiflora, respectively. Serological investigations demonstrated significant depletion in triglycerides and elevation in blood sodium level in animals treated with essential oils of A. marmelos and C. reticulata Blanco. Boenninghausenia albiflora affected many markers including RBC, MCV, triglycerides, HDL, LDL, urea, and sodium. In conclusion, all oils except B. albiflora can be considered safe for internal use.

Antilisterial Effect of Rosa damascena and Nymphaea alba in Mus musculus.

The present study was proposed to investigate the toxicological and prophylactic potential of ethanolic extracts of Rosa damascena and Nymphaea alba and their mixture in albino mice. For toxicity study, three different doses of plant extracts were orally administrated to three groups of mice for 14 successive days. Blood biochemistry and histological examinations of liver and kidney revealed that these extracts had no harmful effects up to 1000 mg/kg. To determine the prophylactic effects of Rosa damascena, Nymphaea alba, and their mixture, an infection model of Listeria monocytogenes was established in a pilot study. Establishment of infection was confirmed by changes in haematological parameters and reisolation of Listeria monocytogenes from different tissues. Results showed that these extracts alone or in combination could restrict the growth of Listeria monocytogenes in different organs. Neutrophils were high in positive control group but remained in normal range in all treated groups. Listeria monocytogenes was recovered in low numbers from animals treated with extract of single plant but was negligible in group treated with mixture of extract of plants. Platelets count was increased in treated groups as compared to control. Results confirmed that these extracts are potent source of antimicrobial compounds and that they have synergistic effect in combined form.

In vivo screening and evaluation of four herbs against MRSA infections.

BACKGROUND: Recently, we reported high in vitro antibacterial efficacy of Althaea officinalis, Ziziphus jujuba, Cordia latifolia and Thymus vulgaris out of a total 21 plants against wide range of bacteria including MRSA. This study was therefore, designed to confirm efficacy of these four herbs against MRSA in an animal model. METHODS: A pilot study was conducted to establish the dose of S. aureus (KY698020) required to induce clinical infection. Afterword, in main trial, efficacy of aforementioned plant extracts on the course of sore throat was checked by evaluating general health, gross lesion score, bacterial load and hematology in mice. RESULTS: Pilot study revealed that 40 µl dose of 107 CFU/ml could induce infection which persist upto 08 days post infection. Mice treated with T. vulgaris and Z. jujuba showed reduction in gross lesion score of both heart and lungs. Treatment with only some plants could significantly decrease bacterial load of throat (T. vulgaris) heart, blood and joint (C. latifolia, and T. vulagris). Hematological indicators confirmed in vivo control of MRSA infection in all treatment groups except A. officinalis. CONCLUSION: This is first report confirming in vivo anti-MRSA potential of C. latifolia and T. vulgaris and highlight the need to explore bioactive constituents of these plants. Moreover, previously reported in vitro antibacterial efficiency of A. officinalis could not be validated in current study.

Phytochemical, Antimicrobial, and Toxicological Evaluation of Traditional Herbs Used to Treat Sore Throat.

The in vitro antibacterial activities of 29 traditional medicinal plants used in respiratory ailments were assessed on multidrug resistant Gram-positive and Gram-negative bacteria isolated from the sore throat patients and two reference strains. The methanolic, n-hexane, and aqueous extracts were screened by the agar well diffusion assay. Bioactive fractions of effective extracts were identified on TLC coupled with bioautography, while their toxicity was determined using haemolytic assay against human erythrocytes. Qualitative and quantitative phytochemical analysis of effective extracts was also performed. Methanolic extract of 18 plants showed antimicrobial activity against test strains. Adhatoda vasica (ZI = 17-21 mm, MIC: 7.12-62.5 µg/mL), Althaea officinalis (ZI = 16-20 mm, MIC: 15.62-31.25 µg/mL), Cordia latifolia (ZI = 16-20 mm, MIC: 12.62-62.5 µg/mL), Origanum vulgare (ZI = 20-22 mm, MIC: 3-15.62 µg/mL), Thymus vulgaris (ZI = 21-25 mm, MIC: 7.81-31.25 µg/mL), and Ziziphus jujuba (ZI = 14-20 mm, MIC: 7.81-31.25 µg/mL) showed significant antibacterial activity. Alkaloid fractions of Adhatoda vasica, Cordia latifolia, and Origanum vulgare and flavonoid fraction of the Althaea officinalis, Origanum vulgare, Thymus Vulgaris, and Ziziphus jujuba exhibited antimicrobial activity. Effective plant extracts show 0.93-0.7% erythrocyte haemolysis. The results obtained from this study provide a scientific rationale for the traditional use of these herbs and laid the basis for future studies to explore novel antimicrobial compounds.

Effect of Peganum harmala or its beta-carboline alkaloids on certain antibiotic resistant strains of bacteria and protozoa from poultry.

In the present study the antimicrobial potential of various extracts from 12 medicinal plants has been investigated in vitro on multiple antibiotic resistant pathogens and some selected protozoa isolated from poultry. The initial examination was performed on E. coli (n = 10) using disc and agar well diffusion assays. Only two plants, Peganum harmala (seeds) and Eucalyptus globulus (leaves) showed positive responses. The active extracts were also investigated against an additional 19 bacteria and the clonal cultures of three protozoa (Histomonas meleagridis, Tetratrichomonas gallinarum and Blastocystis sp.). Only Peganum harmala was found to inhibit the growth of all bacteria and protozoa at 0.38-1.55 mg/mL and 0.63-1.65 mg/mL, respectively. To investigate the potential role of alkaloids in crude extracts of Peganum harmala, four known beta-carbolin alkaloids were quantified and their antimicrobial activity was compared using a microdilution assay. Harmaline was found to be in the highest concentration followed by harmine and harmalol, whereas harmane could not be detected. The activity of the pure alkaloids was in the order harmane > harmaline > harmalol > or = harmine for all bacteria, while for protozoa, it was different depending on the microorganism. It is concluded that Peganum harmala or its alkaloids could probably be used for the control of antibiotic resistant isolates of bacteria as well as protozoa.

Antiprotozoal activities determined in vitro and in vivo of certain plant extracts against Histomonas meleagridis, Tetratrichomonas gallinarum and Blastocystis sp.

A total of 43 plant substances provided as raw material and different kinds of extracts (aqueous, ethanol, and heptane) from 18 different organic wastes obtained from the food/feed industry were investigated for their in vitro activities against clonal cultures of Histomonas meleagridis, Tetratrichomonas gallinarum, and Blastocystis sp. Ethanolic extracts of thyme, saw palmetto, grape seed, and pumpkin fruit proved to be most efficacious. Thus, these extracts were further tested in vivo in turkeys experimentally infected with H. meleagridis by administrating the substances to the birds through the drinking water. Even though a delayed mortality was noticed in some birds medicated with the extracts of thyme, grape seed, and pumpkin fruit, all birds died or had to be euthanized the latest within 5 weeks post infection--with the exception of one bird which was probably never infected with histomonads--due to a severe typhlohepatitis indicative for histomonosis. In addition, none of the substances were able to prevent the spreading of H. meleagridis from infected to in-contact birds. Thus, these studies clearly demonstrate that in vitro studies are of limited value to assess the efficacy of plant substances against histomonosis.

Differences in the in vitro susceptibility of mono-eukaryotic cultures of Histomonas meleagridis, Tetratrichomonas gallinarum and Blastocystis sp. to natural organic compounds.

Currently, all pharmaceuticals for the treatment or prophylaxis of blackhead disease (histomonosis) caused by the flagellate Histomonas meleagridis are banned from the market. Consequently, great interest exists on the finding of alternative drugs for the abatement of histomonosis. In this study, carvacrol, Cassia oil, an essential oil (EO) mixture containing thyme and rosemary EO and a Quillaja saponaria saponin were examined using in vitro assays for antiprotozoal and antibacterial activity testing established against cloned xenic cultures of different isolates of Histomonas meleagridis, Tetratrichomonas gallinarum and Blastocystis sp. Whereas similar minimal lethal concentrations (MLCs) of five Histomonas isolates were obtained for both carvacrol and the EO mixture as well as for the saponin, significantly different MLCs were observed for them with Cassia oil, ranging from 0.25 up to 0.50 microl/ml. Testing the Blastocystis isolates, different MLCs were obtained for all substances, whereas the Tetratrichomonas gallinarum isolates showed identical susceptibilities. The effects are independent of the bacteria, underlining the need of well-defined protozoan cultures for these investigations.