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.

Role of MyD88-Adaptor-Like (MAL) Gene Polymorphism rs8177374 and Cytokine (IFN-γ, TNF-α, IL-10, TGF-ß) Levels in Diverse Malaria Manifestations upon P. falciparum and P. vivax Infections.

This study investigated the role of genetic variant rs8177374 in MAL/TIRAP gene in mediating the cytokine levels of IFN-γ, TNF-α, IL-10, and TGF-ß in malaria patients due to Plasmodium falciparum or P. vivax infection. The study included human blood samples collected from patients with malaria (n = 228) and healthy controls (n = 226). P. falciparum and P. vivax groups were established based on the causative species of Plasmodium. Malaria samples were divided into mild and severe malaria groups based on the symptoms that appeared in the patients, according to the WHO criteria. In a previous study, we genotyped rs8177374 via allele specific PCR strategy. In this study, cytokine levels were estimated in the blood plasma of rs8177374 genotype samples via Sandwich Enzyme Linked Immunosorbent Assay kits. Increased IFN-γ and TNF-α levels in presence of CC genotype indicates the role of CC genotype in both severe and mild malaria groups. Enhanced IL-10 levels in the CT genotype and mild malaria groups suggest a role of CT genotype and IL-10 in the mild clinical outcomes of malaria. The rs8177374 polymorphism in MAL/TIRAP plays an important role in malaria pathogenesis.

Genetic diversity, cholesterol reduction, and presence of conserved bile salt hydrolase gene in probiotic strains from human milk.

Probiotics are known to possess strain- and species-specific functional properties, of which hypocholesteremia is of major interest. Bile salt hydrolase (BSH) activity is one of the key mechanisms involved in the hypocholesterolemic effect. The study was designed to genetically characterize probiotics obtained from human milk on the basis of simple sequence repeat (SSR), isolate potent hypocholesterolemic strains, and detect BSH activity, deconjugation of bile salts, and bsh polymorphism. This study, for the first time, linked genetic diversity with cholesterol reduction potential and proved the presence of conserved bsh of Levilactobacillus brevis in genetically diverse species. The strains displayed 2.78%-42.23% cholesterol reduction, which was not influenced by prebiotics. In this study, data obtained from SSR markers indicated 93.3% diversity, and based on cluster analysis, they were distributed into XI clades; out of five potent cholesterol-reducing strains, three belonged to clade I. The strains could deconjugate both sodium glycocholate and sodium taurocholate, but we preferred using sodium glycocholate. The variation in cholesterol reduction potential and BSH activity pointed toward the presence of more than one bsh in the strains. Weissella confusa MW051433 displayed highest cholesterol reduction (42.23%) and specific BSH activity (2.64 U ml -1). Search for other bsh and in vivo assessments of cholesterol reduction by W. confusa MW051433 have been proposed.

Lead biosorption efficiency of Levilactobacillus brevis MZ384011 and Levilactobacillus brevis MW362779: A response surface based approach.

Lead (Pb) is a substantial contaminant in the environment and a potent toxin for living organisms. Current study describes probiotic characteristics of Pb-biosorbing lactic acid bacteria (LAB), and response surface methodology (RSM) based optimization of physical conditions for maximum Pb biosorption. A total of 18 LAB, isolated from carnivore feces (n = 8) and human breast milk (n = 9), along with one reference strain Lactobacillus acidophilus ATCC4356 were included in the study. Pb biosorption was strain specific. Eight strains, demonstrating ≥ 70 % lead biosorption, were selected for further testing. The lactobacillus-Pb complex was found to be stable and strains had a negative surface charge. The strains displayed good probiotic properties with the survival rate of 71-90 % in simulated gastric environment, 36-69 % in intestinal condition (1.8 % bile salts) and 55-72 % hydrophobicity. On the basis of excellent probiotic ability, Levilactobacillus brevis MZ384011 and Levilactobacillus brevis MW362779 were selected for optimization of physical conditions of Pb biosorption through RSM. Maximum biosorption was observed at pH 6 in 60 min at a cell density of 1 g/L. L. brevis MZ384011 and L. brevis MW362779 are recommended for experimentation on Pb toxicity amelioration and safety evaluation in in-vivo setting.

Levilactobacillus brevis from carnivores can ameliorate hypercholesterolemia: In vitro and in vivo mechanistic evidence.

AIMS: The aim was to explore the probiotic and hypocholesterolaemic potential of two Levilactobacillus brevis strains of carnivore origin along with selected underlying mechanisms. METHODS AND RESULTS: Levilactobacillus brevis MT950194 and L. brevis MW365351 were analysed in vitro for oro-gastro-intestinal stress tolerance, cholesterol reduction, cholesterol adsorption (through scanning electron microscopy) and bile salt hydrolase (BSH) activity. Strains could survive (>80%) in oro-gastro-intestinal conditions and reduce high amount of cholesterol (35% and 54%) from media containing bile salts (0.3%) as compared with Lactobacillus acidophilus ATCC 4356 and presented the least pathogenicity towards mammalian cells. Exopolysaccharide production, cell surface cholesterol adherence and BSH activity were witnessed as possible cholesterol-lowering mechanisms. In in vivo experiment, the treatments of hypercholesterolaemic rats with L. brevis MT950194, L. brevis MW365351 and their mixture led to significant (p < 0.05) reduction in serum and hepatic cholesterol, low-density lipids, cholesterol ratio, liver steatosis and size of adipocytes. It further ameliorated diet-induced changes in hepatic enzymes. CONCLUSIONS: Levilactobacillus brevis MT950194 and L. brevis MW365351 from carnivores have probiotic pharmacological potential and can reduce serum cholesterol through surface adherence and BSH production. SIGNIFICANCE AND IMPACT OF THE STUDY: These strains may be utilized in treating hypercholesterolaemia and production of low-fat functional foods.

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.

Evaluation of inhibitory and probiotic properties of lactic acid bacteria isolated from vaginal microflora.

Lactic acid bacteria (LABs) are known to secrete species-specific secondary metabolites that could be utilized as novel therapeutics against multi-drug resistant pathogens. This study aimed to investigate the antagonistic and probiotic properties of LABs isolated from the vaginal ecosystem of healthy women and to assess the stability of their antagonistic metabolites. Among 43 strains isolated from healthy women, eight LAB strains exhibited detectable BLISs (bacteriocin-like substances) producing ability against E. faecalis (JH-86), S. aureus (JH-68), Streptococcus sp. (JH-80), and E. coli (JH-101), with zone of inhibition (ZI) ranging from 9.00 to 20.33 mm and minimum inhibitory concentrations (MICs) from 62.5 to 500 µL/mL, respectively. The partially purified compounds extracted from cell free supernatant (CFS) displayed an increase in antagonistic activity based on ZI, 9.67-30.17 mm and MICs, 3.91-15.63 mg/mL, respectively. In a time-kill study, both crude and partially purified compounds of Limosilactobacillus reuteri (MT180537), Pediococcus pentosaceus (MT176555), Limosilactobacillus pontis (MW362838), and Levilactobacillus brevis (MW362790) exhibited significant bactericidal action against E. faecalis (MW051601), the most frequent etiological agent of aerobic vaginitis (AV). The active secondary metabolites from L. reuteri (MT180537), P. pentosaceus (MT176555), and L. pontis (MW362838) were protein in nature and remained stable under different physicochemical conditions. Regarding probiotic properties, the strains presented probiotic characteristics, i.e., good acid, bile salt tolerance, aggregation properties, and biofilm formation. The strains were susceptible to most of the commonly used antibiotics and had no hemolytic activity. In conclusion, antagonistic compounds or BLIS produced by L. reuteri (MT180537) could be investigated further for preparation of ointments to treat AV.

In vivo prophylactic efficacy of Lactobacillus reuteri MT180537 against aerobic vaginitis.

Aerobic vaginitis is a recently described vaginal infection that is treated with antibiotics, which cause undesirable effects leading to disturbance in normal vaginal flora and antibiotic resistance among pathogens. Probiotics may be considered as a natural alternative therapy. We investigated antagonistic and immunomodulatory potential of intravaginally administered probiotic Lactobacillus reuteri-MT180537 against vaginal colonization by Enterococcus faecalis-MW051601 in mice. In vitro antimicrobial potential of lactic acid bacteria was determined against major pathogens of aerobic vaginitis. Moreover, in vivo prophylactic efficacy of L. reuteri-MT180537 against E. faecalis-MW051601 induced AV, in ß-estradiol immunosuppressed mice was determined for the first time. Lactic acid bacteria displayed antibacterial activity against pathogens with zone of inhibition (11.33-20.00 mm) and co-aggregation (40-67%). Animals receiving L. reuteri-MT180537 followed by E. faecalis-MW051601 challenge exhibited significant reduction in clinical index, vaginal bacterial load, and histopathological changes in vaginal tissues compared to animals receiving E. faecalis-MW051601 only. L. reuteri-MT180537 upregulated expression of anti-inflammatory (Foxp3, IFN-γ) cytokines and resulted in controlling E. faecalis-MW051601 induced over expression of pro-inflammatory (IL-6, IL-1ß) cytokines. Altogether, L. reuteri-MT180537 displayed antagonistic properties in vitro and prevented aerobic vaginitis by inhibiting the growth of E. faecalis-MW051601 and regulating expression of pro-inflammatory and anti-inflammatory cytokines in mice.

Multienzyme and Antibacterial Potential of Bacteria Isolated from gut of Asian Honey Bee (Apis cerana Indica), Lahore Using Culture Dependent Method

Abstract The bacteria residing in the gut of honey bees (HB) has demonstrated a significant role in protecting bees against various pathogens, production of honey and wax. However, no information exists about the antibacterial potential of bacterial isolates from gut of Asian HB, Apis cerana Indica F. (Hymenoptera: Apidae), against human pathogens. This study aims to investigate the antibacterial and multienzyme potential of aerobic bacteria from A. cerana gut using culture dependent approach. A total of 12 HB gut bacteria were characterized morphologically and biochemically. These strains were further screened for their antimicrobial activity against pathogenic human microorganisms Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Bacillus licheniformis and Bacillus subtilis using cross streak (primary screening) and agar well diffusion methods (secondary screening). Preliminary characterization of cell-free supernatant (CFS) of two promising isolates was performed by measuring lactic acid concentrations, enzymatic digestion of antimicrobial compounds, stability over a range of temperature, pH and amplification of spaS (subtilin) and spoA (subtilosin) genes. In primary screening, among 12 HB isolates, eight strains showed statistically significant highest zones of inhibition (p≤0.05) against E. coli, K. pneumoniae and P. aeruginosa. 16S rRNA sequencing revealed that these isolates belong to Bacillus genus, identified as B. tequilensis, B. pumilus, B. xiamenensis, B. subtilis, B. amyloliquefaciens, B. safensis, B. licheniformis, B. altitudinis (Accession numbers: MT186230-MT186237). Secondary screening revealed that among eight isolates, B. subtilis and B. amyloliquefaciens showed statistically significantly strong inhibition (p≤0.05) against all tested pathogens. Antibiotic susceptibility testing revealed that both isolates were resistant to antibiotics and possesses proteolytic, lipolytic and cellulolytic activities. The nature of the compound causing inhibitory activity was found to be proteinaceous and showed stability over a wide range of temperature as well as pH. PCR study confirmed the presence of bacteriocins by successful amplification of important antimicrobial peptide biosynthesis genes spaS and spoA. These results suggest that the HB gut is a home to bacteria that possess antimicrobial activity and important enzymes with antimicrobial potential. To our knowledge, this is the first report demonstrating the antimicrobial potential of bacteria isolated from gut of HB (A. cerana) against human pathogens.

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.

Motility effects biofilm formation in Pseudomonas aeruginosa and Enterobacter cloacae.

Chronic infections caused by gram negative bacteria are the mains reasons to have morbidity and death in patients, despite using high doses of antibiotics applied to cure diseases producing by them. This study was designed to identify the role of flagella in biofilm formation Ten pure strains were collected from our lab. Morphological variation and motility assays led us to study two strains in detail. They were characterized biochemically, physiologically and genetically. Biofilm formation analysis was performed using test tube assay, congo red assay and liquid-interface coverslip assay. In order to disrupt flagella of studied strains, blending was induced for 5, 10 and 15 minutes followed by centrifugation and observing motility using motility test. Biofilm quantification of wild type (parental) and blended strains was done using test tube and liquid interface coverslip assays. 16S rRNA sequencing identified strains as Pseudomonas aeroginosa and Enterobacter cloacae. Significant biofilm formation (p>0.05) by was observed after 72 and 18 hours using test tube and liquid-interface coverslip assays respectively. Flagellar disruption showed that 15 minutes blending caused significant reduction in both strains, hence demonstrated that flagellar mediated motility could be a potent strategy to stabilize aggregate and invest resources for biofilm formation in P. aeruginosa and E. cloacae.

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.

Identification of virulence factors in contact lens associated bacteria: A physiological approach.

PURPOSE: Wearing contact lens requires awareness about possible contaminants, the causative agents of multiple complications. The present study focused on identification of potential pathogens and presence of virulence associated markers in contact lens associated bacteria. METHODS: Bacterial contaminants were isolated from contact lenses or cleaning solutions collected from University students. Isolates were identified using conventional methods followed by 16S rRNA gene sequencing and screened for the presence of virulence factors which included capsular presence, adhesion, serum resistance, iron chelation, haemagglutination and hemolysis. Moreover, antibiotic resistance profile was also monitored. RESULTS: Contamination was observed in 79% (45 of 57) of lenses. Based on 16S rRNA sequencing Bacillus sp. was found to be most abundant (26%). The presence of at least three pathogenic characteristics was recorded in 75.8% isolates. Among the pathogenic characteristics, capsule presence was found to be the most prevalent character (73%) followed by hemolysin production (65%), serum resistance (61%), haemagglutination (56%), iron chelation (50%) and polystyrene adherence (42%). Multiple antibiotic resistance was recorded in 66.13% isolates. Cluster analysis on the basis of virulence markers separated all isolates in two groups. Potential pathogens and non-pathogens were found to be equally frequent among contaminants of contact lens cases. CONCLUSION: The present work provides evidence that pathogenic bacteria can adhere and survive in contact lens or lens solution. It highlights the need for the development of new methods to protect contact lenses and lens care accessories. Drugs targeting capsule formation may offer a good option for treatment or use in cleaning solution.

Assessment of the risk for human health of Enterovirus and Hepatitis A virus in clinical and water sources from three metropolitan cities of Pakistan.

INTRODUCTION: Molecular studies have confirmed the silent circulation of enterovirus (EntV) and hepatitis A virus in the environment, even in the absence of clinical manifestation. Viral pathogens are among the major causes of disease outbreaks, particularly in the bigger cities and both in the developed and underdeveloped nations. MATERIAL AND METHODS: Between June 2016 - June 2017, 97 samples of drinking water, river water polluted with sewage and blood were selected and obtained from high risk communities in Pakistan. Negatively charged membrane filters were used to concentrate the virus, followed by the use of specific PCR primers set for quick identification of the waterborne viruses. RESULTS: Enteroviruses were recovered from 40%, 28.57% and 33.33% of river water polluted with sewage samples in Lahore, Islamabad and Rawalpindi, respectively, while the presence of 13.13% and 11.76% of viral load was also confirmed in the drinking water of Lahore and Rawalpindi, respectively. A high prevalence of HAV (12.5% and 21.05%) was also verified in the clinical samples. Phylogenetic analysis indicated close resemblance of HAV isolates with the Indian strains. This study is the first ever comparative analysis of the EntV and HAV isolated from environmental samples and clinical specimen on a molecular level. CONCLUSIONS: The parallel surveillance of EntV and HAV in the river water polluted with sewage, and clinical samples is quite helpful for controlling and reducing the disease burden of the waterborne illnesses.

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.

Association between Genetic Polymorphism and Risk of von Willebrand Disease in Pakistan.

von Willebrand disease (VWD) is an inherited, genetically and clinically heterogeneous hemorrhagic disorder. The most common cause of this disease is mutation in the gene that encodes protein von Willebrand factor (VWF) which is responsible for blood clotting. The current study was designed to investigate the role of genetic polymorphisms with the onset of VWD in population of Pakistan. Three exonic variants (c.3445T>C; c.4975C>T; c.7603C>T) from VWF gene were used for the genotyping purpose. The current study employed a case-control association design involving 43 VWD patients and 100 healthy controls from Pakistani population. The genetic reason of VWD was investigated using the allele specific PCR. The significant (P < 0.05) allelic association was found between all three exonic variants and VWD. The CT genotype of these variants was noticed to be associated with significantly higher risk of VWD [odds ratio (95% CI): 14.7 (4.546-47.98), 26.71 (7.281-97.98), and 21.5 (5.806-80.01) for c.3445T>C, c.4975C>T, and c.7603C>T, resp.] while genotypes CC (c.4975C>T) and TT (c.3445T>C and c.7603C>T) were having protective effect against the disease. However, replicated studies are needed for elaborating the role of these SNPs.

Prevalence of rotavirus, adenovirus, hepatitis A virus and enterovirus in water samples collected from different region of Peshawar, Pakistan.

Viral gastroenteritis and other water-borne diseases are the most neglected areas of research in Pakistan. To determine the quality of water, 4 enteric viruses were studied from different localities of Peshawar, Pakistan. The study validates the viral detection method for Rotavirus (RV), Human adenovirus (HAdV), Enterovirus (EV) and Hepatitis A virus (HAV), directly from water sources of rural areas of Peshawar, KPK, Pakistan. Overall, 95 five water samples were tested; among them, 9.47% were positive for RV, 38.94% for HAdV, 48.42% for EV and 12.63% for HAV. The presence of these viruses in water was directly correlated with meteorological data. High prevalence of EV and HAdV was detected frequently in the wet season from May - September, which can be the potential cause of spreading of gastroenteritis in the population. Environmental surveillance is an additional tool to evaluate the epidemiology of enteric viruses circulating in a given community.

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.