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Sample records for lawsonia intracellularis serpulina

  1. The seroprevalence of Lawsonia intracellularis in horses in The Netherlands.

    PubMed

    Kranenburg, L C; van Ree, H E M I; Calis, A N M; de Pater, M; Buter, G J; van Maanen, C; Sloet van Oldruitenborgh-Oosterbaarn, M M

    2011-04-01

    Equine proliferative enteropathy caused by Lawsonia intracellularis is an emerging disease of weanling foals and affects their growth and development. The prevalence of Lawsonia intracellularis in The Netherlands is not known. The aim of the study was to investigate the seroprevalence of Lawsonia intracellularis in horses in The Netherlands. Blood samples were taken from healthy foals before and after weaning and from healthy yearlings and mature horses on farms throughout The Netherlands. These samples were analysed for the presence of Lawsonia intracellularis-specific antibodies with a blocking ELISA. White blood cell count, packed cell volume, and total protein concentration were also measured in all foals. Information regarding housing, pasture access, and contact with pig manure on the premises was obtained for all animals. The prevalence of Lawsonia intracellularis antibodies in foals increased significantly from 15% before weaning to 23% after weaning (p = 0.019); it was 89% in yearlings and 99% in horses older than 2 years. There was no significant difference in seroprevalence between the pasture-kept and stable-confined adult horses (97% and 100%, respectively), and there was no significant influence of contact with pig manure. None of the sampled animals showed clinical disease. In conclusion, the results suggest that Lawsonia intracellularis is widespread in The Netherlands and that seropositivity is not necessarily associated with clinical problems. The high seroprevalence in adult horses suggests long-term persistence of antibodies against Lawsonia intracellularis or constant exposure to the bacterium.

  2. Lawsonia intracellularis and Porcine Circovirus type-2 infection in Estonia.

    PubMed

    Järveots, T; Saar, T; Põdersoo, D; Rüütel-Boudinot, S; Sütt, S; Tummeleht, L; Suuroja, T; Lindjärv, R

    2016-01-01

    The present study describes the reasons of post-weaning distress in Estonian pig herds. Here we examined the natural cases of Lawsonia intracellularis and porcine circovirus 2 (PCV2) infection and co-infections. The presence of L. intracellularis in swine herds were tested by PCR and by histopathological methods, whereas PCV2 was detected by real-time-PCR and immunohistochemical stainings. Seven of the 11 investigated herds with signs of post-weaning wasting were infected with L. intracellularis and all 11 herds with PCV2. From the analysed samples 22.2% were infected with L. intracellularis and 25% with PCV2. The results of microbiological studies suggested that the piglets suffered from enteritis and pneumonia. Escherichia coli and Pasteurella multocida often aggravated the process of illness. The frequency of L. intracellularis was high in pigs 7-12 weeks old (18.5-42.7%) and PCV2 infection was too high in pigs 7-12 weeks old (24.8-32.7%). E. coli was often a co-factor with L. intracellularis and PCV2. The primary reasons of post weaning wasting were PCV2 and E. coli, later aggravated by L. intracellularis and other pathogens. Our results indicated that different pathogens have an important role in developing post-weaning wasting. Proliferative intestinal inflammation caused by L. intracellularis is mainly characterised by its localization and morphological findings. The main gross lesions were the enlargement of mesenteric lymph nodes and thickening of the wall of ileum. In post-weaning multi-systemic wasting syndrome there are characteristic histological lesions in lymphoid tissues. They consist of a variable degree of lymphocyte depletion, together with histiocytic and/or multinucleate giant cell infiltration. This basic lymphoid lesions is observable in almost all tissues of a single severely affected animal, including lymph nodes, Peyer's patches and spleen. Sporadically, multifocal coagulative necrosis may be observed. PMID:27487502

  3. A Novel Lawsonia intracellularis Autotransporter Protein Is a Prominent Antigen▿

    PubMed Central

    Watson, Eleanor; Clark, Ewan M.; Alberdi, M. Pilar; Inglis, Neil F.; Porter, Megan; Imrie, Lisa; Mclean, Kevin; Manson, Erin; Lainson, Alex; Smith, David G. E.

    2011-01-01

    Investigation of antigenic determinants of the microaerophilic obligate intracellular bacterium Lawsonia intracellularis using a mass spectrometry approach identified a novel bacterial protein present in an extract of cell culture medium aspirated from heavily infected in vitro cell cultures. Western immunoblotting analysis of SDS-PAGE-resolved proteins using immune sera pooled from L. intracellularis-infected pigs revealed the presence of a strongly immunoreactive band of ∼72 kDa. Liquid chromatography-electrospray ionization-tandem mass spectrometry analysis of this component and database mining using a fully annotated L. intracellularis genome sequence and the comprehensive GenBank prokaryotic genomic database highlighted the presence of a protein that shares little sequence similarity with other prokaryotic proteins and appears to be highly species specific. Detailed bioinformatic analyses identified the protein as member of the autotransporter protein family of surface-exposed proteins, and the designation LatA (Lawsonia autotransporter protein A) is suggested. Recognition of recombinant LatA on Western blots by a panel of sera from infected and control pigs corresponded 100% with a commercial serodiagnostic that relies on in vitro culture of this fastidious organism. LatA therefore represents a potential candidate for the development of a rapid and species-specific serodiagnostic reagent. PMID:21697340

  4. Lawsonia intracellularis-associated ulcerative and necro-hemorrhagic enteritis in 5 weanling foals.

    PubMed

    Arroyo, Luis G; Ter Woort, Federica; Baird, John D; Tatiersky, Laetitia; Delay, Josepha; van Dreumel, Tony

    2013-09-01

    This report describes 5 cases of fatal Lawsonia intracellularis-associated ulcerative and necro-hemorrhagic enteritis in weanling Thoroughbred and Standardbred foals. The lesions are similar to those of the L. intracellularis-associated ulcerative and necro-hemorrhagic enteritis syndrome in pigs. Two foals had concurrent severe typhlo-colitis as a result of a large burden of encysted cyathostomes. The clinical, diagnostic, and therapeutic challenges, and the potential complications encountered during the management of such cases are discussed.

  5. Transmission of Lawsonia intracellularis to weanling foals using feces from experimentally infected rabbits.

    PubMed

    Pusterla, N; Sanchez-Migallon Guzman, D; Vannucci, F A; Mapes, S; White, A; DiFrancesco, M; Gebhart, C

    2013-02-01

    The aim of this study was to investigate whether feces from rabbits experimentally infected with Lawsonia intracellularis were infectious to foals. Two rabbits were infected with L. intracellularis, while two rabbits served as controls. Eight foals received daily feces from either the infected or the control rabbits. All rabbits and foals were monitored daily for clinical signs for the entire study period (21days for rabbits, 42days for foals). Feces and blood were collected for the PCR detection of L. intracellularis and serologic analysis, respectively. None of the infected rabbits or foals developed clinical signs compatible with proliferative enteropathy. All infected rabbits and foals shed L. intracellularis in their feces and all seroconverted. The results support the role of rabbits as asymptomatic amplifiers of L. intracellularis and their role as sources of infection for susceptible foals.

  6. Proteomic analysis of Lawsonia intracellularis reveals expression of outer membrane proteins during infection.

    PubMed

    Watson, Eleanor; Alberdi, M Pilar; Inglis, Neil F; Lainson, Alex; Porter, Megan E; Manson, Erin; Imrie, Lisa; Mclean, Kevin; Smith, David G E

    2014-12-01

    Lawsonia intracellularis is the aetiological agent of the commercially significant porcine disease, proliferative enteropathy. Current understanding of host-pathogen interaction is limited due to the fastidious microaerophilic obligate intracellular nature of the bacterium. In the present study, expression of bacterial proteins during infection was investigated using a mass spectrometry approach. LC-ESI-MS/MS analysis of two isolates of L. intracellularis from heavily-infected epithelial cell cultures and database mining using fully annotated L. intracellularis genome sequences identified 19 proteins. According to the Clusters of Orthologous Groups (COG) functional classification, proteins were identified with roles in cell metabolism, protein synthesis and oxidative stress protection; seven proteins with putative or unknown function were also identified. Detailed bioinformatic analyses of five uncharacterised proteins, which were expressed by both isolates, identified domains and motifs common to other outer membrane-associated proteins with important roles in pathogenesis including adherence and invasion. Analysis of recombinant proteins on Western blots using immune sera from L. intracellularis-infected pigs identified two proteins, LI0841 and LI0902 as antigenic. The detection of five outer membrane proteins expressed during infection, including two antigenic proteins, demonstrates the potential of this approach to interrogate L. intracellularis host-pathogen interactions and identify novel targets which may be exploited in disease control.

  7. In vitro antimicrobial activity against 10 North American and European Lawsonia intracellularis isolates.

    PubMed

    Wattanaphansak, Suphot; Singer, Randall S; Gebhart, Connie J

    2009-03-01

    The objective of this study was to determine the in vitro minimum inhibitory concentration (MIC) of antimicrobials against 10 isolates of Lawsonia intracellularis, the etiological agent of proliferative enteropathy (PE). Antimicrobials tested included carbadox, chlortetracycline, lincomycin, tiamulin, tylosin and valnemulin. The MIC of each antimicrobial against L. intracellularis was determined using a tissue culture system and was identified as the lowest concentration that inhibited 99% of L. intracellularis growth, as compared to the antimicrobial-free control. Each antimicrobial concentration was evaluated for both intracellular and extracellular activity against L. intracellularis, an obligately intracellular bacterium. When tested for intracellular activity, carbadox, tiamulin, and valnemulin were the most active antimicrobials with MICs of < or =0.5microg/ml. Tylosin (MICs ranging from 0.25 to 32microg/ml) and chlortetracycline (MICs ranging from 0.125 to 64microg/ml) showed intermediate activities and lincomycin (MICs ranging from 8 to >128mIcog/ml) showed the least activity. When tested for extracellular activity, valnemulin (MICs ranging from 0.125 to 4microg/ml) was the most active against most L. intracellularis isolates. Chlortetracycline (MICs ranging from 16 to 64microg/ml), tylosin (MICs ranging from 1 to >128microg/ml), and tiamulin (MICs ranging from 1 to 32microg/ml) showed intermediate activities. Lincomycin (MICs ranging from 32 to >128microg/ml) showed the least activity. Our in vitro results showed that each L. intracellularis isolate had a different antimicrobial sensitivity pattern and these data can be utilized as an in vitro guideline for the further antimicrobial evaluation of field L. intracellularis isolates. PMID:18823723

  8. Efficacy of gallium maltolate against Lawsonia intracellularis infection in a rabbit model.

    PubMed

    Sampieri, F; Allen, A L; Alcorn, J; Clark, C R; Vannucci, F A; Pusterla, N; Mapes, S M; Ball, K R; Dowling, P M; Thompson, J; Bernstein, L R; Gebhart, C J; Hamilton, D L

    2014-12-01

    Antimicrobial efficacy against Lawsonia intracellularis is difficult to evaluate in vitro, thus, the effects of gallium maltolate's (GaM) were investigated in a rabbit model for equine proliferative enteropathy (EPE). Juvenile (5-6-week-old) does were infected with 3.0 × 10(8) L. intracellularis/rabbit and allocated into three groups (n = 8). One week postinfection, one group was treated with GaM, 50 mg/kg; one, with doxycycline, 5 mg/kg; and one with a sham-treatment (control). Feces and blood were collected daily and weekly, respectively, to verify presence of L. intracellularis fecal shedding using qPCR, and seroconversion using immunoperoxidase monolayer assay. Rabbits were sacrificed after 1 week of treatment to collect intestinal tissues focusing on EPE-affected sections. Intestinal lesions were confirmed via immunohistochemistry. No difference was noted between treatments regarding EPE-lesions in jejunum (P = 0.51), ileum (P = 0.74), and cecum (P = 0.35), or in L. intracellularis fecal shedding (P = 0.64). GaM and doxycycline appear to have similar efficacy against EPE in infected rabbits.

  9. Porcine proliferative enteropathy in Estonian pig herds: histopathology and detection of Lawsonia intracellularis by PCR.

    PubMed

    Järveots, Tönu; Saar, Tiiu; Lepp, Elbi; Suuroja, Toivo; Lindjärv, Raivo; Nathues, Heiko; Sütt, Silva; Põdersoo, Diivi

    2011-01-01

    Lawsonia intracellularis is the causative agent of proliferative enteritis in pigs (PPE). This bacterium is difficult to culture from clinical samples and antemortem demonstration is therefore usually performed by PCR on faecal samples. The aim of this study was to elucidate the frequency of L. intracellularis infection in pig herds in Estonia using PCR, histopathological methods and electronmicroscopical studies. The frequency of demonstration of L. intracellularis was highest in 9-12 weeks old pigs (68.1%). It was more frequent in growing pigs with enteritis on small farms where the system of "all-in all-out" was not practiced and where standards of hygiene were poor. Gross and histopathological studies demonstrated that characteristic macroscopic changes associated with PPE were localised to the distal jejunum and ileum.Thickened longitudinal and circumferential folds occurred in the mucosa of the affected regions of the bowel. Samples from pigs aged 4 to 20 weeks exhibited the most intensive inflammatory changes. The distal part of the jejunum, ileum and the upper third of proximal colon and cecum wall were visibly thickened with reduced luminal diameter. Hyperplasia of lymphoid tissue and, in many cases, pseudomembranous or fibrinous inflammation was found. L. intracellularis was detected in 56 young pigs using histopathological methods. Additionally, in 8 of these pigs intracellular bacteria were demonstrated in ilial epithelial cells by transmission electronmicroscopical (TEM) investigation. On the basis of these TEM investigations it was concluded that L. intracellularis causes disturbances of normal growth, differentiation and apoptosis of the epithelial cells of ileum. PMID:21306056

  10. Further investigation of exposure to Lawsonia intracellularis in wild and feral animals captured on horse properties with equine proliferative enteropathy.

    PubMed

    Pusterla, Nicola; Mapes, Samantha; Gebhart, Connie

    2012-11-01

    This study investigated the exposure to Lawsonia intracellularis in wild birds, mice, rabbits, raccoons, coyotes and squirrels, and feral cats and pigs on 10 farms with confirmed equine proliferative enteropathy (EPE). Serum samples from all resident foals (417 samples) as well as fecal (461) and serum (106) samples from wild and feral animals were collected for serological and molecular detection of L. intracellularis following the diagnosis of EPE in index cases. A total of three cats from two farms, three mice from two farms and eight cottontail rabbits from one farm had evidence of prior exposure to L. intracellularis. These animals may be an indicator of environmental exposure or may be actively involved in the transmission of L. intracellularis to foals by acting as a potential reservoir/amplifier host.

  11. Cytologic diagnosis of Lawsonia intracellularis proliferative ileitis in a Japanese snow macaque (Macaca fuscata).

    PubMed

    Wamsley, Heather L; Wellehan, James F X; Harvey, John W; Embury, Jennifer E; Troutman, J Mitchell; Lafortune, Maud

    2005-01-01

    Diffuse ileal thickening and ileocecocolic lymphadenomegaly were observed during exploratory laparotomy in a 2-year-old male Japanese snow macaque (Macaca fuscata) that had flu-like signs and diarrhea. Cytologic examination of ileal biopsy imprints revealed many mature, mildly karyolytic neutrophils and fewer well-differentiated lymphocytes, eosinophils, macrophages, and plasma cells in a background containing amorphous, necrotic material. Tightly cohesive sheets of moderately pleomorphic epithelial cells also were seen. The cytologic diagnosis was chronic, active, mixed inflammation with atypical epithelial cells and necrosis. Histologically, the mucosal and crypt epithelium was moderately hyperplastic with a loss of goblet cells, increased mitoses, and frequent crypt abscesses. Within the lamina propria and extending into the submucosa was a marked neutrophilic infiltrate, with low numbers of lymphocytes, histiocytes, plasma cells, and eosinophils. The histologic diagnosis was chronic, diffuse, marked suppurative and lymphocytic ileitis. Warthin-Starry silver staining of the ileal biopsy and imprint specimens demonstrated numerous pleomorphic, curved bacilli consistent with Lawsonia intracellularis. Polymerase chain reaction (PCR) and immunohistochemistry confirmed the identity of the infectious agent. L intracellularis infection may be underdiagnosed because silver stain is required to visualize the organism with light microscopy and because the pathognomonic crypt hyperplasia may be complicated by secondary pathologic changes. Application of silver stain to cytologic specimens should be considered when distal intestinal lesions associated with hyperplastic epithelium, with or without inflammation, hemorrhage, or necrosis, are identified in animals with clinical signs of enteritis, especially in frequently affected species or in stressed or young animals.

  12. Species-specificity of equine and porcine Lawsonia intracellularis isolates in laboratory animals

    PubMed Central

    Sampieri, Francesca; Vannucci, Fabio A.; Allen, Andrew L.; Pusterla, Nicola; Antonopoulos, Aphroditi J.; Ball, Katherine R.; Thompson, Julie; Dowling, Patricia M.; Hamilton, Don L.; Gebhart, Connie J.

    2013-01-01

    Lawsonia intracellularis infection causes proliferative enteropathy (PE) in many mammalian species, with porcine and equine proliferative enteropathy (PPE and EPE) known worldwide. Hamsters are a well-published animal model for PPE infection studies in pigs. There is no laboratory animal model for EPE infection studies and it is not known whether there is species-specificity for equine or porcine isolates of L. intracellularis in animal models. The objective of this study was to determine whether it is possible to generate typical EPE lesions in hamsters after inoculation with an equine strain of L. intracellularis (EPE strain) and whether it is comparatively possible to generate PPE lesions in rabbits after inoculation with a porcine strain of L. intracellularis (PPE strain). In 2 separate trials, 4-week-old and 3-week-old weanling golden Syrian hamsters were challenged with EPE strains and compared to uninfected (both trials) and PPE-infected controls (Trial 2 only). Concurrently, 6 female New Zealand white juvenile rabbits were infected with PPE strain and observed concomitantly to 8 similar rabbits infected with EPE strain for a different experiment. Hamsters and rabbits were observed for 21 to 24 days post-infection (DPI), depending on the experiment. Neither infected species developed clinical signs. The presence of disease was assessed with diagnostic techniques classically used for pigs and horses: immune-peroxidase monolayer assay on sera; quantitative polymerase chain reaction (qPCR) detection of molecular DNA in feces; and hematoxylin and eosin (H&E) stain and immunohistochemistry (IHC) on intestinal tissues. Our results showed that EPE-challenged hamsters do not develop infection when compared with PPE controls (IHC, P = 0.009; qPCR, P = 0.0003). Conversely, PPE-challenged rabbits do not develop typical intestinal lesions in comparison to EPE-challenged rabbits, with serological response at 14 DPI being significantly lower (P = 0.0023). In conclusion

  13. Species-specificity of equine and porcine Lawsonia intracellularis isolates in laboratory animals.

    PubMed

    Sampieri, Francesca; Vannucci, Fabio A; Allen, Andrew L; Pusterla, Nicola; Antonopoulos, Aphroditi J; Ball, Katherine R; Thompson, Julie; Dowling, Patricia M; Hamilton, Don L; Gebhart, Connie J

    2013-10-01

    Lawsonia intracellularis infection causes proliferative enteropathy (PE) in many mammalian species, with porcine and equine proliferative enteropathy (PPE and EPE) known worldwide. Hamsters are a well-published animal model for PPE infection studies in pigs. There is no laboratory animal model for EPE infection studies and it is not known whether there is species-specificity for equine or porcine isolates of L. intracellularis in animal models. The objective of this study was to determine whether it is possible to generate typical EPE lesions in hamsters after inoculation with an equine strain of L. intracellularis (EPE strain) and whether it is comparatively possible to generate PPE lesions in rabbits after inoculation with a porcine strain of L. intracellularis (PPE strain). In 2 separate trials, 4-week-old and 3-week-old weanling golden Syrian hamsters were challenged with EPE strains and compared to uninfected (both trials) and PPE-infected controls (Trial 2 only). Concurrently, 6 female New Zealand white juvenile rabbits were infected with PPE strain and observed concomitantly to 8 similar rabbits infected with EPE strain for a different experiment. Hamsters and rabbits were observed for 21 to 24 days post-infection (DPI), depending on the experiment. Neither infected species developed clinical signs. The presence of disease was assessed with diagnostic techniques classically used for pigs and horses: immune-peroxidase monolayer assay on sera; quantitative polymerase chain reaction (qPCR) detection of molecular DNA in feces; and hematoxylin and eosin (H&E) stain and immunohistochemistry (IHC) on intestinal tissues. Our results showed that EPE-challenged hamsters do not develop infection when compared with PPE controls (IHC, P = 0.009; qPCR, P = 0.0003). Conversely, PPE-challenged rabbits do not develop typical intestinal lesions in comparison to EPE-challenged rabbits, with serological response at 14 DPI being significantly lower (P = 0.0023). In conclusion

  14. Changes in the Porcine Intestinal Microbiome in Response to Infection with Salmonella enterica and Lawsonia intracellularis.

    PubMed

    Borewicz, Klaudyna A; Kim, Hyeun Bum; Singer, Randall S; Gebhart, Connie J; Sreevatsan, Srinand; Johnson, Timothy; Isaacson, Richard E

    2015-01-01

    Salmonella enterica is a leading cause of food borne illness. Recent studies have shown that S. enterica is a pathogen capable of causing alterations to the composition of the intestinal microbiome. A recent prospective study of French pork production farms found a statistically significant association between Lawsonia intracellularis and carriage of S. enterica. In the current study the composition of the gut microbiome was determined in pigs challenged with S. enterica serovar Typhimurium and or L. intracellularis and compared to non-challenged control pigs. Principal coordinate analysis demonstrated that there was a disruption in the composition of the gut microbiome in the colon and cecum of pigs challenged with either pathogen. The compositions of the microbiomes of challenged pigs were similar to each other but differed from the non-challenged controls. There also were statistically significant increases in Anaerobacter, Barnesiella, Pediococcus, Sporacetigenium, Turicibacter, Catenibacterium, Prevotella, Pseudobutyrivibrio, and Xylanibacter in the challenged pigs. To determine if these changes were specific to experimentally challenged pigs, we determined the compositions of the fecal microbiomes of naturally infected pigs that were carriers of S. enterica. Pigs that were frequent shedders of S. enterica were shown to have similar fecal microbiomes compared to non-shedders or pigs that shed S. enterica infrequently. In a comparison of the differentially abundant bacteria in the naturally infected pigs compared to experimentally challenged pigs, 9 genera were differentially abundant and each exhibited the same increase or decrease in abundance between the two groups. Thus, there were similar changes in the GI microbiome associated with carriage of S. enterica regardless of whether the pigs were experimentally challenged with S. enterica or acquired it naturally.

  15. Changes in the Porcine Intestinal Microbiome in Response to Infection with Salmonella enterica and Lawsonia intracellularis

    PubMed Central

    Singer, Randall S.; Gebhart, Connie J.; Sreevatsan, Srinand; Johnson, Timothy; Isaacson, Richard E.

    2015-01-01

    Salmonella enterica is a leading cause of food borne illness. Recent studies have shown that S. enterica is a pathogen capable of causing alterations to the composition of the intestinal microbiome. A recent prospective study of French pork production farms found a statistically significant association between Lawsonia intracellularis and carriage of S. enterica. In the current study the composition of the gut microbiome was determined in pigs challenged with S. enterica serovar Typhimurium and or L. intracellularis and compared to non-challenged control pigs. Principal coordinate analysis demonstrated that there was a disruption in the composition of the gut microbiome in the colon and cecum of pigs challenged with either pathogen. The compositions of the microbiomes of challenged pigs were similar to each other but differed from the non-challenged controls. There also were statistically significant increases in Anaerobacter, Barnesiella, Pediococcus, Sporacetigenium, Turicibacter, Catenibacterium, Prevotella, Pseudobutyrivibrio, and Xylanibacter in the challenged pigs. To determine if these changes were specific to experimentally challenged pigs, we determined the compositions of the fecal microbiomes of naturally infected pigs that were carriers of S. enterica. Pigs that were frequent shedders of S. enterica were shown to have similar fecal microbiomes compared to non-shedders or pigs that shed S. enterica infrequently. In a comparison of the differentially abundant bacteria in the naturally infected pigs compared to experimentally challenged pigs, 9 genera were differentially abundant and each exhibited the same increase or decrease in abundance between the two groups. Thus, there were similar changes in the GI microbiome associated with carriage of S. enterica regardless of whether the pigs were experimentally challenged with S. enterica or acquired it naturally. PMID:26461107

  16. Lawsonia intracellularis infection of intestinal crypt cells is associated with specific depletion of secreted MUC2 in goblet cells

    PubMed Central

    Bengtsson, Rebecca J.; MacIntyre, Neil; Guthrie, Jack; Wilson, Alison D.; Finlayson, Heather; Matika, Oswald; Pong-Wong, Ricardo; Smith, Sionagh H.; Archibald, Alan L.; Ait-Ali, Tahar

    2015-01-01

    The expression patterns of secreted (MUC2 and MUC5AC) and membrane-tethered (MUC1, MUC4, MUC12 and MUC13) mucins were monitored in healthy pigs and pigs challenged orally with Lawsonia intracellularis. These results showed that the regulation of mucin gene expression is distinctive along the GI tract of the healthy pig, and may reflect an association between the function of the mucin subtypes and different physiological demands at various sites. We identified a specific depletion of secreted MUC2 from goblet cells in infected pigs that correlated with the increased level of intracellular bacteria in crypt cells. We concluded that L. intracellularis may influence MUC2 production, thereby altering the mucus barrier and enabling cellular invasion. PMID:26377360

  17. Comparative genome sequencing identifies a prophage-associated genomic island linked to host adaptation of Lawsonia intracellularis infections

    PubMed Central

    2013-01-01

    Lawsonia intracellularis is an obligate intracellular bacterium and the causative agent of proliferative enteropathy (PE). The disease is endemic in pigs, emerging in horses and has also been reported in a variety of other animal species, including nonhuman primates. Comparing the whole genome sequences of a homologous porcine L. intracellularis isolate cultivated for 10 and 60 passages in vitro, we identified a 18-kb prophage-associated genomic island in the passage 10 (pathogenic variant) that was lost in the passage 60 (non-pathogenic variant). This chromosomal island comprises 15 genes downstream from the prophage DLP12 integrase gene. The prevalence of this genetic element was evaluated in 12 other L. intracellularis isolates and in 53 infected animals and was found to be conserved in all porcine isolates cultivated for up to 20 passages and was lost in isolates cultivated for more than 40 passages. Furthermore, the prophage region was also present in 26 fecal samples derived from pigs clinically affected with both acute and chronic forms of the disease. Nevertheless, equine L. intracellularis isolates evaluated did not harbor this genomic island regardless of the passage in vitro. Additionally, fecal samples from 21 clinically affected horses and four wild rabbits trapped in horse farms experiencing PE outbreaks did not show this prophage-associated island. Although the presence of this prophage-associated island was not essential for a virulent L. intracellularis phenotype, this genetic element was porcine isolate-specific and potentially contributed to the ecological specialization of this organism for the swine host. PMID:23826661

  18. Laser microdissection coupled with RNA-seq analysis of porcine enterocytes infected with an obligate intracellular pathogen (Lawsonia intracellularis)

    PubMed Central

    2013-01-01

    Background Lawsonia intracellularis is an obligate intracellular bacterium and the etiologic agent of proliferative enteropathy. The disease is endemic in pigs, emerging in horses and has been described in various other species including nonhuman primates. Cell proliferation is associated with bacterial replication in enterocyte cytoplasm, but the molecular basis of the host-pathogen interaction is unknown. We used laser capture microdissection coupled with RNA-seq technology to characterize the transcriptional responses of infected enterocytes and the host-pathogen interaction. Results Proliferative enterocytes was associated with activation of transcription, protein biosynthesis and genes acting on the G1 phase of the host cell cycle (Rho family). The lack of differentiation in infected enterocytes was demonstrated by the repression of membrane transporters related to nutrient acquisition. The activation of the copper uptake transporter by infected enterocytes was associated with high expression of the Zn/Cu superoxide dismutase by L. intracellularis. This suggests that the intracellular bacteria incorporate intracytoplasmic copper and express a sophisticated mechanism to cope with oxidative stress. Conclusions The feasibility of coupling microdissection and RNA-seq was demonstrated by characterizing the host-bacterial interactions from a specific cell type in a heterogeneous tissue. High expression of L. intracellularis genes encoding hypothetical proteins and activation of host Rho genes infers the role of unrecognized bacterial cyclomodulins in the pathogenesis of proliferative enteropathy. PMID:23800029

  19. First molecular identification of Entamoeba polecki in a piglet in Japan and implications for aggravation of ileitis by coinfection with Lawsonia intracellularis.

    PubMed

    Matsubayashi, Makoto; Kanamori, Kenta; Sadahiro, Masayuki; Tokoro, Masaharu; Abe, Niichiro; Haritani, Makoto; Shibahara, Tomoyuki

    2015-08-01

    Parasitic Entamoeba spp. are found in many vertebrate species including humans, as well as many livestock including pigs. In pigs, three Entamoeba spp., E. suis, and E. polecki and E. histolytica as zoonotic species, have been identified, but their pathogenicity has not been fully characterized. Here, we report the bacteriological, virological, and histopathological examination of three piglets with chronic diarrhea. Two animals appeared to be additionally infected with Lawsonia intracellularis, which caused a characteristic proliferative ileitis. In the piglet infected with Entamoeba spp., the trophozoites (approximately 10-15 μm with one nucleus in their cytoplasm) invaded into the lamina propria and the disease was worsened by the formation of ulcers and pseudomembranes. Genetic analysis identified the parasite as E. polecki (99.5% identity). Although E. polecki in humans or animals might be less pathogenic in the case of a single infection, coinfections with other pathogens including L. intracellularis may increase the severity of the disease.

  20. The efficacy of oxytetracycline treatment at batch, pen and individual level on Lawsonia intracellularis infection in nursery pigs in a randomised clinical trial.

    PubMed

    Larsen, Inge; Nielsen, Søren Saxmose; Olsen, John Elmerdahl; Nielsen, Jens Peter

    2016-02-01

    Antimicrobial consumption in animal husbandry is of great scientific and political concern due to the risk of selection of resistant bacteria. Whilst a reduction in the use of antimicrobials is therefore preferable, the efficacy of treatment must be maintained in order to ensure animal welfare and profitability of pig production. The objective of this study was to evaluate the efficacy of three treatment strategies under field conditions against Lawsonia intracellularis (LI)-related diarrhoea. A randomised clinical trial was carried out in four Danish pig herds, including a total of 520 pigs from 36 nursery batches. A high prevalence of LI was demonstrated in all herds prior to the initiation of the study. Treatment efficacy was assessed by faecal shedding of LI, the occurrence of diarrhoea and average daily weight gain (ADG) after treatment. All strategies were implemented at batch level at presence of LI-related diarrhoea and included daily treatment with 10mg oxytetracycline (OTC) per kilogram of bodyweight for 5 days, though the OTC was administered differently: either by oral treatment of all pigs in a batch, by oral treatment of pigs in diarrhoeic pens only, or by intramuscular treatment of individual diarrhoeic pigs only. The treatment strategies were randomly allocated to batches and were initiated at the presence of diarrhoea. From the included batches, 100% of the trial pigs were medicated in the batch treatment strategy, 87% in the pen treatment strategy and 55% in the individual treatment strategy. All strategies reduced the occurrence of diarrhoea and faecal shedding of LI after treatment. However, batch treatment was found to be most efficient in reducing both high-level LI shedding and diarrhoea when compared to the treatment of diarrhoeic pens or individual diarrhoeic pigs. There was no significant difference identified in ADG between the treatment strategies. In conclusion, batch treatment of all pigs in a section resulted in the highest efficacy

  1. Restriction fragment length polymorphism of the periplasmic flagellar flaA1 gene of Serpulina species.

    PubMed Central

    Fisher, L N; Mathiesen, M R; Duhamel, G E

    1997-01-01

    Forty-one reference and field isolates of intestinal spirochetes representing Serpulina hyodysenteriae, Serpulina innocens, Serpulina pilosicoli, Brachyspira aalborgi, and nonclassified weakly beta-hemolytic intestinal spirochetes were compared by restriction fragment length polymorphism (RFLP) of the periplasmic flagellar (PF) flaA1 gene. Six genetically distinct groups (I through VI), each with a unique RFLP fingerprint pattern, were identified by Southern blotting analysis of EcoRV chromosomal DNA digests with a PCR-amplified digoxigenin-labeled 1-kb fragment of the S. hyodysenteriae isolate B78 PF flaA1 gene. The RFLP fingerprint patterns corresponded to known DNA homology differences between Serpulina species and to provisionally designated species described previously by using phenotypic and genotypic classification schemes. RFLP fingerprinting of the PF flaA1 gene provides a relatively simple genotypic method for identification of intestinal spirochetes without the use of radioisotopes. PMID:9384289

  2. Pharmacokinetics of gallium maltolate in Lawsonia intracellularis-infected and uninfected rabbits.

    PubMed

    Sampieri, F; Alcorn, J; Allen, A L; Clark, C R; Vannucci, F A; Pusterla, N; Mapes, S; Ball, K R; Dowling, P M; Thompson, J; Bernstein, L R; Gebhart, C J; Hamilton, D L

    2014-10-01

    Oral gallium maltolate (GaM) pharmacokinetics (PK) and intestinal tissue (IT) concentrations of elemental gallium ([Ga]) and iron ([Fe]) were investigated in a rabbit model of equine proliferative enteropathy (EPE). New Zealand white does (uninfected controls and EPE-infected, n = 6/group) were given a single oral GaM dose (50 mg/kg). Serial blood samples were collected from 0 to 216 h post-treatment (PT) and IT samples after euthanasia. Serology, qPCR, and immunohistochemistry confirmed, or excluded, EPE. Blood and IT [Ga] and [Fe] were determined using inductively coupled plasma-mass spectrometry. PK parameters were estimated through noncompartmental approaches. For all statistical comparisons on [Ga] and [Fe] α = 5%. The Ga log-linear terminal phase rate constant was lower in EPE rabbits vs. uninfected controls [0.0116 ± 0.004 (SD) vs. 0.0171 ± 0.0028 per hour; P = 0.03]; but half-life (59.4 ± 24.0 vs. 39.4 ± 10.8 h; P = 0.12); Cmax (0.50 ± 0.21 vs. 0.59 ± 0.42 μg/mL; P = 0.45); tmax (1.75 ± 0.41 vs. 0.9 ± 0.37 h; P = 0.20); and oral clearance (6.743 ± 1.887 vs. 7.208 ± 2.565 L/h; P = 0.74) were not. IT's [Ga] and [Fe] were higher (P < 0.0001) in controls. In conclusion, although infection reduces IT [Ga] and [Fe], a 48 h GaM dosing interval is appropriate for multidose studies in EPE rabbits.

  3. Identification and characterization of genus-specific epitopes of Serpulina species using monoclonal antibodies.

    PubMed

    Achacha, M; Mittal, K R

    1996-01-01

    Four murine monoclonal antibodies (mAbs) designated as C9E8, A10, G12, and G8 which recognized both Serpulina hyodysenteriae and S. innocens were produced and characterized. The mAbs reacted with whole cell antigens in ELISA, indirect immunofluorescence and immunoblot assays. The mAbs did not show any cross reactivity in rapid dot ELISA or immunoblot assay with Leptospira icterohemorrhagiae, Campylobacter jejuni and Escherichia coli. Treatment of whole cell suspension with proteinase K and sodium periodate indicated that the reacting epitopes of the mAbs were protein in nature. The genus-specific antigens were identified as heat-stable proteins with molecular weight in the range of 26 to 45 kDa. Immunofluorescence and immunogold labelling studies showed that the antibody-binding epitopes were exposed on the outer-surface of the spirochaetal cell wall. The mAbs inhibited growth of reference strains of both S. hyodysenteriae and S. innocens in vitro but failed to cause agglutination. The detection of spirochaetal forms directly in fecal smears or paraffin-embeded tissue sections from experimentally infected pigs indicated that such mAbs were potentially useful for the diagnosis of swine spirochaetosis. This is the first report of mAbs identifying and characterizing common antigens of porcine Serpulina.

  4. Production and characterization of monoclonal antibodies against Serpulina hyodysenteriae and S. innocens and their use in serotyping.

    PubMed Central

    Achacha, M; Mittal, K R

    1995-01-01

    Murine monoclonal antibodies (MAbs) directed against serotypes 1, 2, 8, and 9 of Serpulina hyodysenteriae and strain B256 of Serpulina innocens were produced and characterized. A serological classification of 96 field strains of S. hyodysenteriae and 28 field strains of S. innocens isolated from pigs showing clinical signs of swine dysentery was performed by rapid dot enzyme-linked immunosorbent assay (ELISA) with the MAbs. The results indicated that the majority of the field strains of S. hyodysenteriae (69%) belonged to serotypes 8, 1, and 9, whereas only 31% of the S. innocens strains were recognized by MAb 9H7, indicating the presence of antigenic heterogeneity among S. innocens isolates. Rapid dot ELISA with type-specific MAbs was found to be specific, sensitive, and easy to perform and thus to be suitable for routine serotyping of S. hyodysenteriae and S. innocens isolates. This is the first report of MAbs being used for serotyping clinical isolates of S. hyodysenteriae and S. innocens. PMID:7494066

  5. Pigs experimentally infected with Serpulina hyodysenteriae can be protected from developing swine dysentery by feeding them a highly digestible diet.

    PubMed Central

    Siba, P. M.; Pethick, D. W.; Hampson, D. J.

    1996-01-01

    Weaner pigs (n = 72) were fed 1 of 4 diets. These were based on either cooked rice and animal protein, cooked rice and lupin, wheat and lupin, or wheat and animal protein. Twenty-six of the pigs were slaughtered after 1 month. Those fed the highly digestible cooked rice and animal protein diet had drier colonic contents and faeces, lighter large intestines, and the contents of their large intestines had increased pH values and decreased total VFA concentrations. The other 46 were orally challenged with broth cultures of Serpulina hyodysenteriae, and were monitored for faecal excretion of the spirochaetes, and for the development of swine dysentery (SD). None of 18 pigs fed the cooked rice and animal protein diet developed colonic changes or disease, whereas most pigs on the other diets developed mucohaemorrhagic colitis and dysentery. The reduced fermentation that occurred in the large intestines of pigs fed cooked rice and animal protein was associated with a subsequent failure of colonization by S. hyodysenteriae, and resultant protection against SD. PMID:8620913

  6. Enzyme-linked immunosorbent assay for measuring ileal symbiont intracellularis-specific immunoglobulin G response in sera of pigs.

    PubMed Central

    Holyoake, P K; Cutler, R S; Caple, I W; Monckton, R P

    1994-01-01

    Proliferative enteritis (PE) is a common intestinal disease on pig farms. The disease is caused by ileal symbiont (IS) intracellularis (Campylobacter-like organisms) bacteria. An enzyme-linked immunosorbent assay (ELISA) was developed to measure IS intracellularis-specific immunoglobulin G (IgG) response in the sera of pigs. The antigen used in the ELISA was filtered, percoll gradient-purified IS intracellularis extracted from the intestines of pigs affected with proliferative hemorrhagic enteropathy. The antibody responses of pigs challenged with intestinal homogenates from pigs affected with proliferative hemorrhagic enteropathy containing IS intracellularis or percoll-gradient purified IS intracellularis were low and variable. The low IgG titers measured in challenged pigs support previous findings that IgG plays a minor role in the immune response of pigs to IS intracellularis. On a farm in which infection was endemic, pigs seroconverted at between 7 and 24 weeks of age. High IgG titers, indicative of maternally acquired antibody, were present in 3-week-old pigs. The IgG titers in piglets were lowest at 6 weeks of age, which approximates the age of onset of clinical disease. These results suggest that IgG plays a role in determining the susceptibilities of pigs to natural infection. Measurements of seroconversion by the ELISA might aid in epidemiological investigations of PE in naturally infected herds. However, the variable antibody responses in experimentally challenged pigs would seem to limit its usefulness as an antemortem diagnostic test for PE. PMID:7989553

  7. Comparison between the effect of Lawsonia inermis and flubendazole on Strongyloides species using scanning electron microscopy.

    PubMed

    Ismail, Khadiga Ahmed; Ibrahim, Ayman Nabil; Ahmed, Mona Abdel-Fattah; Hetta, Mona Hafez

    2016-06-01

    Strongyloides species is a helminth of worldwide distribution primarily in tropical and subtropical regions. It is the only soil-transmitted helminth with the ability for autoinfection so; it may lead to severe systemic manifestations especially in immunosuppressed patients. Chemotherapy is currently considered the best therapeutic option for strongyloidiasis but some drugs are expensive and others have side effects as nausea, diarrhea and headache. Strongyloides larva is resistant to most chemical agents so, search for plant extracts may provide other effective but less expensive treatment. Lawsonia inermis Linn, popularly known as Henna, has been proven to have antihelminthic, antibacterial and antifungal properties. The current study was carried out to evaluate the efficacy of Lawsonia inermis on Strongyloides spp. In vitro using scanning electron microscopy. Fifty Strongyloides species. larvae and free living females were incubated with different concentrations of Lawsonia (1, 10, 100 mg/ml), for different incubation periods (24, 48, 72 and 96 h) in comparison to the same concentrations of flubendazole at the same different time points. The results showed that Lawsonia inermis in a concentration of 10 mg/ml incubated with Strongyloides spp. female for 24 h affected the parasite cuticular surface in the form of transverse and longitudinal fissures and transverse depression in comparison to no cuticular change with flubendazole (100 mg/ml). This suggests that Lawsonia inermis may be a promising phytotherapeutic agent for strongyloidiasis. PMID:27413314

  8. Molluscicidal activity of Lawsonia inermis and its binary and tertiary combinations with other plant derived molluscicides.

    PubMed

    Singh, A; Singh, D K

    2001-03-01

    Molluscicidal activity of leaf, bark and seed of Lawsonia inermis against Lymnaea acuminata and Indoplanorbis exustus was studied. Highest toxicity was observed in the seed of Lawsonia inermis. Toxicity of binary (1:1) and tertiary (1:1:1) combinations of the essential oil of cedar (Cedrus deodara Roxh) and neem (Azadirachta indica A. Juss), powder from bulb of garlic (Allium sativum Linn), and oleoresin extracted from rhizome of ginger (Zingiber officinale Rosc) with Lawsonia inermis and Embelia ribes fruit powder were studied against L. acuminata and I. exustus. L. inermis seed powder in combination with Cedrus deodara oil and Azadirachta indica oil was more toxic than their individual components and other combinations. PMID:11495286

  9. Phytochemical, toxicological and antimicrobial evaluation of lawsonia inermis extracts against clinical isolates of pathogenic bacteria

    PubMed Central

    2013-01-01

    Background The emerging resistance of pathogen against the currently available antimicrobial agents demands the search of new antimicrobial agents. The use of medicinal plants as natural substitute is the paramount area of research to overwhelm the drug resistance of infectious agents. Scientists have not made enough effort on the evaluation of safety of medicinal plant yet. Methods In the present study antimicrobial activity of Lawsonia inermis is investigated against clinical isolates of seven bacteria including four Gram negative (Escherichia coli, Salmonella typhi, Klebsiella spp., Shigella sonnei) and three Gram positive (Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis) using disc diffusion method. Four types of Lawsonia inermis extracts were prepared using methanol, chloroform, acetone and water as extraction solvents, while DMSO (Dimethyl sulfoxide) and water as dissolution solvents. The rate and extent of bacterial killing was estimated by time-kill kinetic assay at 1× MIC of each bacterial isolate. The overall safety of Lawsonia inermis extracts was assessed in mice. Results Lawsonia inermis displayed noteworthy antimicrobial activity against both gram positive and gram negative bacterial strains used in the study. The minimum value of MIC for different bacterial strains ranged from 2.31 mg/ml to 9.27 mg/ml. At 1x MIC of each bacterial isolate, 3log10 decrease in CFU was recorded after 6 hours of drug exposure and no growth was observed in almost all tested bacteria after 24 hours of exposure. No sign of toxidrome were observed during in vivo toxicity evaluation in mice at 300 mg/kg concentration. Conclusion In conclusion, the present study provides the scientific rational for medicinal use of Lawsonia inermis. The use of Lawsonia inermis extracts is of great significance as substitute antimicrobial agent in therapeutics. PMID:24289297

  10. Micropropagation of commercially cultivated Henna (Lawsonia inermis) using nodal explants.

    PubMed

    Ram, Kheta; Shekhawat, N S

    2011-07-01

    Lawsonia inermis Linn. (Mehandi) is cultivated as cash crop in India particularly in Sojat area of Pali district, Rajasthan. Present investigation describes an efficient regeneration system for elite genotype of L. inermis using nodal segments. Optimum response in terms of percent cultures responding, days to bud break and average shoot length was observed on MS medium supplemented with 6-benzylaminopurine (BA; 2.0 mg l(-1)). Shoot multiplication was influenced by plant growth regulators, repeated transfer of explants and addition of ammonium sulphate. Maximum shoots were regenerated on MS medium supplemented with BA (0.25 mg l(-1)), kinetin (Kn; 0.25 mg l(-1)), indole-3-acetic acid (IAA; 0.1 mg l(-1)) and ammonium sulphate (150 mg l(-1)). To reduce resources, time and labours costs, we have also attempted ex vitro rooting of shoots. About 95 % shoots were rooted ex vitro on soilrite after treatment with indole-3-butyric acid (IBA; 300 mg l(-1)) and 2-naphthoxy acetic acid (NOA; 100 mg l(-1)) and establishment in soil successfully.

  11. Antibacterial activity of Phyllantus emblica, Coriandrum sativum, Culinaris medic, Lawsonia alba and Cucumis sativus.

    PubMed

    Khan, Dawood Ali; Hassan, Fouzia; Ullah, Hanif; Karim, Sabiha; Baseer, Abdul; Abid, Mobasher Ali; Ubaidi, Muhammad; Khan, Shujaat Ali; Murtaza, Ghulam

    2013-01-01

    Present study deals with the demonstration of the antibacterial activity of very common medicinal plants of Pakistani origin i.e., Phyllantus emblica, Coriandrum sativum, Culinaris medic, Lawsonia alba and Cucumis sativus. The extracts were prepared in crude form by the use of hydro-alcoholic solution and were screened for antibacterial activity against various bacterial species by disk diffusion method. Assay was performed using clinical isolates of B. cereus, S. aureus, P. aeruginosa and E. coli. Crude extract of Phyllantus emblica fruit exhibited strong activity against standard cultures of all studied bacteria. Lawsonia alba showed good activity against standard cultures of all the used microorganisms. Coriandrum sativum was effective only against Bacillus cereus, while Cucumis sativus and Culinaris medic showed poor activity against Pseudomonas aeruginosa only. Hence, Phyllantus emblica exhibited strong antibacterial activity against a wide range of bacteria it means that Phyllantus emblica extract contains some compounds which have broad spectrum of bactericidal activity.

  12. The Abortificient Effects of Hydroalcoholic Extract of Lawsonia Inermis on BALB/c Mice

    PubMed Central

    Esteki, Ramin; Miraj, Sepideh

    2016-01-01

    Introduction According to the traditional beliefs of certain cultures, Lawsonia inermis has been reported to cause the abortion or termination of an undesirable pregnancy. The present study was undertaken with the goal of studying the effect of Lawsonia inermis extract on abortion in pregnant BALB/c mice in 2013 in Shahrekord, Iran. Methods This research study used an experimental methodology and was conducted in 2013 in Shahrekord, Iran. Forty female BALB/c mice (30–40 gm, 8–12 weeks old) were randomly assigned to 4 groups. One male mouse was included for each two female mice (1:2) and they were maintained in a protective cage habitat. Pregnancy of the mice was confirmed by means of a vaginal smear. The doses of 1 mg/kg and 10 mg/kg of the hydroalcoholic extract of Lawsonia inermis were injected intraperitoneally into pregnant mice beginning on the first day and continuing through the seventh day of pregnancy. The control group did not receive any treatment, but was left completely unadministered. On the eighteenth day of pregnancy, the uterine tubes of mice were removed. The subsequent embryonic absorption is considered to be an abortion. The data were analyzed using SPSS software version 22 using Fisher’s exact test and the Kruskal-Wallis H tests. Results Abortions were observed more often in the experimental groups (p< 0.01). The mean of the serum estrogen level was significantly higher in the case control groups (p< 0.01) and the mean of progesterone level was also significantly lower in the experimental groups (p< 0.01). Conclusion The use of Lawsonia inermis during pregnancy may cause abortion and therefore it should be considered as contraindication or use with caution. PMID:27504174

  13. Antileishmanial activity of some plants growing in Algeria: Juglans regia, Lawsonia inermis and Salvia officinalis.

    PubMed

    Serakta, M; Djerrou, Z; Mansour-Djaalab, H; Kahlouche-Riachi, F; Hamimed, S; Trifa, W; Belkhiri, A; Edikra, N; Hamdi Pacha, Y

    2013-01-01

    The current study was undertaken to evaluate in vitro the antileishmanial activity of three plants growing wild in Algeria : Juglans regia, Lawsonia inermis and Salvia officinalis. The hydroalcoholic extracts of these plants were tested on the growth of the promastigotes of Leishmania major. The plant extract effects were compared with three controls : CRL1 composed of 1 ml RPMI inoculated with 10(6) of promastigotes, CRL2 composed of 1 ml RPMI inoculated with 10(6) of promastigotes and 100 µl of hydroalcoholic solvent, CRL3 composed of 1 ml RPMI inoculated with 10(6) of promastigotes and 100 µl of Glucantim as a reference drug in the management of leishmaniasis. The results showed that both J. regia and L. inermis extracts reduced the promastigotes number significantly (P<0.01). however, S. officinalis showed a total inhibition of the Leishmania major growth.

  14. Antioxidant Activity and Teratogenicity Evaluation of Lawsonia Inermis in BALB/c Mice

    PubMed Central

    Jafarzadeh, Lobat; Seifi, Neda; Shahinfard, Najmeh; Sedighi, Mehrnoosh; Kheiri, Soleiman; Shirzad, Hedayatollah

    2015-01-01

    Background and Aim Lawsonia inermis is a medicinal plant with abortive properties. There has been no scientific study to evaluate the teratogenicity of this plant. This study was performed to determine the antioxidant activity and the possible side effect of L. inermis hydroalcoholic extract on development of congenital abnormalities in BALB/c mice. Materials and Methods In this experimental study, 120 female mature BALB/c mice were assigned to four groups and after mating and confirming the vaginal plug, the animals in the first group (G1) were kept with no intervention, and the second (G2), third (G3) and fourth (G4) groups were intraperitoneally (ip) injected with respectively saline (0.3 ml), and 10 and 100 mg/kg of L. inermis extract (for 7 days). On the 19th day, caesarean section was performed on the mice and embryos were examined for abnormalities. Their height and weight were measured. Data were analysed by ANOVA and post-hoc least significant difference tests. Results There were significant differences between G3 and G4, and G1 (p<0.001); no significant difference was seen between G3 and G4. At 100 mg/kg dose of L. inermis, the parietal bones were absent in 90% of embryos and more extra ribs were observed in both G3 and G4 (p = 0.01). Conclusion L. inermis may have teratogenicity and should be used cautiously during pregnancy. PMID:26155492

  15. Natural dye extract of lawsonia inermis seed as photo sensitizer for titanium dioxide based dye sensitized solar cells

    NASA Astrophysics Data System (ADS)

    Ananth, S.; Vivek, P.; Arumanayagam, T.; Murugakoothan, P.

    2014-07-01

    Natural dye extract of lawsonia inermis seed were used as photo sensitizer to fabricate titanium dioxide nanoparticles based dye sensitized solar cells. Pure titanium dioxide (TiO2) nanoparticles in anatase phase were synthesized by sol-gel technique and pre dye treated TiO2 nanoparticles were synthesized using modified sol-gel technique by mixing lawsone pigment rich natural dye during the synthesis itself. This pre dye treatment with natural dye has yielded colored TiO2 nanoparticles with uniform adsorption of natural dye, reduced agglomeration, less dye aggregation and improved morphology. The pure and pre dye treated TiO2 nanoparticles were subjected to structural, optical, spectral and morphological studies. Dye sensitized solar cells (DSSC) fabricated using the pre dye treated and pure TiO2 nanoparticles sensitized by natural dye extract of lawsonia inermis seed showed a promising solar light to electron conversion efficiency of 1.47% and 1% respectively. The pre dye treated TiO2 based DSSC showed an improved efficiency of 47% when compared to that of conventional DSSC.

  16. Natural dye extract of lawsonia inermis seed as photo sensitizer for titanium dioxide based dye sensitized solar cells.

    PubMed

    Ananth, S; Vivek, P; Arumanayagam, T; Murugakoothan, P

    2014-07-15

    Natural dye extract of lawsonia inermis seed were used as photo sensitizer to fabricate titanium dioxide nanoparticles based dye sensitized solar cells. Pure titanium dioxide (TiO2) nanoparticles in anatase phase were synthesized by sol-gel technique and pre dye treated TiO2 nanoparticles were synthesized using modified sol-gel technique by mixing lawsone pigment rich natural dye during the synthesis itself. This pre dye treatment with natural dye has yielded colored TiO2 nanoparticles with uniform adsorption of natural dye, reduced agglomeration, less dye aggregation and improved morphology. The pure and pre dye treated TiO2 nanoparticles were subjected to structural, optical, spectral and morphological studies. Dye sensitized solar cells (DSSC) fabricated using the pre dye treated and pure TiO2 nanoparticles sensitized by natural dye extract of lawsonia inermis seed showed a promising solar light to electron conversion efficiency of 1.47% and 1% respectively. The pre dye treated TiO2 based DSSC showed an improved efficiency of 47% when compared to that of conventional DSSC.

  17. Inhibitors of melanogenesis in B16 melanoma 4A5 cells from flower buds of Lawsonia inermis (Henna).

    PubMed

    Nakashima, Souichi; Oda, Yoshimi; Nakamura, Seikou; Liu, Jiang; Onishi, Koko; Kawabata, Miki; Miki, Hisako; Himuro, Yugo; Yoshikawa, Masayuki; Matsuda, Hisashi

    2015-07-01

    The methanolic extract of Lawsonia inermis L. (henna) showed significant inhibitory activity toward melanogenesis in B16 melanoma 4A5 cells. Among the constituents isolated from the methanolic extract, luteolin, quercetin, and (±)-eriodictyol showed stronger inhibitory activity than the reference compound, arbutin. Several structure-activity relationships of the flavonoids were suggested, and OGlc

  18. [Hemolytic anemia after voluntary ingestion of henna (Lawsonia inermis) decoction by a young girl with G6PD deficiency].

    PubMed

    Perinet, I; Lioson, E; Tichadou, L; Glaizal, M; de Haro, L

    2011-06-01

    Henna (Lawsonia inermis) is a shrub bearing leaves that are crushed and used for cosmetic purposes in Asia and Africa. In several countries, henna decoction is ingested as a traditional drug to induce abortion. One component of Henna, known as Lawsone, can induce hemolysis in G6PD-deficient patients after cutaneous exposure or ingestion. The purpose of this report is to describe a case of severe hemolytic anemia after voluntary ingestion of Henna decoction to induce abortion. This complication led to diagnosis of partial moderate G6PD-deficiency in the 17-year-old patient living in Mayotte in the Indian Ocean. This report emphasizes the life-threatening hazards associated with some plant extracts used as traditional medicines. PMID:21870562

  19. 'Candidatus Adiutrix intracellularis', an endosymbiont of termite gut flagellates, is the first representative of a deep-branching clade of Deltaproteobacteria and a putative homoacetogen.

    PubMed

    Ikeda-Ohtsubo, Wakako; Strassert, Jürgen F H; Köhler, Tim; Mikaelyan, Aram; Gregor, Ivan; McHardy, Alice C; Tringe, Susannah Green; Hugenholtz, Phil; Radek, Renate; Brune, Andreas

    2016-09-01

    Termite gut flagellates are typically colonized by specific bacterial symbionts. Here we describe the phylogeny, ultrastructure and subcellular location of 'Candidatus Adiutrix intracellularis', an intracellular symbiont of Trichonympha collaris in the termite Zootermopsis nevadensis. It represents a novel, deep-branching clade of uncultured Deltaproteobacteria widely distributed in intestinal tracts of termites and cockroaches. Fluorescence in situ hybridization and transmission electron microscopy localized the endosymbiont near hydrogenosomes in the posterior part and near the ectosymbiont 'Candidatus Desulfovibrio trichonymphae' in the anterior part of the host cell. The draft genome of 'Ca. Adiutrix intracellularis' obtained from a metagenomic library revealed the presence of a complete gene set encoding the Wood-Ljungdahl pathway, including two homologs of fdhF encoding hydrogenase-linked formate dehydrogenases (FDHH ) and all other components of the recently described hydrogen-dependent carbon dioxide reductase (HDCR) complex, which substantiates previous claims that the symbiont is capable of reductive acetogenesis from CO2 and H2 . The close phylogenetic relationship between the HDCR components and their homologs in homoacetogenic Firmicutes and Spirochaetes suggests that the deltaproteobacterium acquired the capacity for homoacetogenesis via lateral gene transfer. The presence of genes for nitrogen fixation and the biosynthesis of amino acids and cofactors indicate the nutritional nature of the symbiosis.

  20. 'Candidatus Adiutrix intracellularis', an endosymbiont of termite gut flagellates, is the first representative of a deep-branching clade of Deltaproteobacteria and a putative homoacetogen.

    PubMed

    Ikeda-Ohtsubo, Wakako; Strassert, Jürgen F H; Köhler, Tim; Mikaelyan, Aram; Gregor, Ivan; McHardy, Alice C; Tringe, Susannah Green; Hugenholtz, Phil; Radek, Renate; Brune, Andreas

    2016-09-01

    Termite gut flagellates are typically colonized by specific bacterial symbionts. Here we describe the phylogeny, ultrastructure and subcellular location of 'Candidatus Adiutrix intracellularis', an intracellular symbiont of Trichonympha collaris in the termite Zootermopsis nevadensis. It represents a novel, deep-branching clade of uncultured Deltaproteobacteria widely distributed in intestinal tracts of termites and cockroaches. Fluorescence in situ hybridization and transmission electron microscopy localized the endosymbiont near hydrogenosomes in the posterior part and near the ectosymbiont 'Candidatus Desulfovibrio trichonymphae' in the anterior part of the host cell. The draft genome of 'Ca. Adiutrix intracellularis' obtained from a metagenomic library revealed the presence of a complete gene set encoding the Wood-Ljungdahl pathway, including two homologs of fdhF encoding hydrogenase-linked formate dehydrogenases (FDHH ) and all other components of the recently described hydrogen-dependent carbon dioxide reductase (HDCR) complex, which substantiates previous claims that the symbiont is capable of reductive acetogenesis from CO2 and H2 . The close phylogenetic relationship between the HDCR components and their homologs in homoacetogenic Firmicutes and Spirochaetes suggests that the deltaproteobacterium acquired the capacity for homoacetogenesis via lateral gene transfer. The presence of genes for nitrogen fixation and the biosynthesis of amino acids and cofactors indicate the nutritional nature of the symbiosis. PMID:26914459

  1. Identification of the gene encoding BmpB, a 30 kDa outer envelope lipoprotein of Brachyspira (Serpulina) hyodysenteriae, and immunogenicity of recombinant BmpB in mice and pigs.

    PubMed

    Lee, B J; La, T; Mikosza, A S; Hampson, D J

    2000-10-01

    A gene encoding a 30kDa outer envelope protein of the intestinal spirochaete Brachyspira (Serpulina) hyodysenteriae, was cloned and expressed in Escherichia coli strain XLOLR. Five phagemids containing DNA inserts encoding the protein were established and one clone (pSHA) was sequenced. An 816bp hypothetical open reading frame (ORF) was identified, with a potential ribosome binding site (AGGAG), and putative -10 (TATAAT) and -35 (TTGAAA) promoter regions upstream from the ATG start of the ORF. A 12bp inverted repeat sequence, possibly serving as a transcription terminator, was identified downstream from the TAA stop codon. Analysis of the amino acid sequence identified a 19 residue hydrophobic signal peptide, incorporating a potential signal peptidase cleavage site and membrane lipoprotein lipid attachment site. Further analysis of the amino acid usage of this lipoprotein, designated BmpB, showed its possible outer membrane localisation. Comparison of the gene encoding the lipoprotein, bmpB, with GenBank nucleotide sequences showed that it has homology with the gene (plp3) encoding Plp3, an outer membrane lipoprotein of Pasteurella haemolytica (54% identity in 735bp). Comparison of the deduced amino acid sequence with the SWISS-PROT amino acid database revealed greatest homology with the outer membrane lipoproteins (Plp1, 2, 3) of P. haemolytica (34% identity in 242 aa, 37% identity in 250 aa, and 39% identity in 272 aa, respectively), and lipoproteins (rcsF and lipoprotein-28) of E. coli (40% identity in 267 aa and 36% identity in 263 aa, respectively). Three of the recombinant E. coli clones (pSHA, pSHD, and pSHE) were formalinised and used to immunise mice. A bacterin preparation of one recombinant E. coli clone (pSHA) was used to immunise pigs. Sera from these mice and pigs recognised the 30kDa lipoprotein in outer membrane preparations of B. hyodysenteriae, indicating the immunogenicity of recombinant BmpB. Sera from pigs naturally infected with B

  2. Effects of Lawsonia inermis L. (Henna) leaves’ methanolic extract on carbon tetrachloride-induced hepatotoxicity in rats

    PubMed Central

    Mohamed, Musab Awad; Eldin, Imad Mohamed Taj; Mohammed, Abd-Elwahab Hassan; Hassan, Hozeifa Mohamed

    2016-01-01

    Background: Natural products with therapeutic properties such as plants, minerals, and animal products, for many years, were the main sources of drugs for the treatment of numerous diseases; hence selection of Lawsonia inermis L. (Henna) to study its hepatoprotective activity was considered. Objectives: This was an attempt to evaluate the hepatoprotective effect of L. inermis leaves’ methanolic extract on carbon tetrachloride (CCl4)-induced hepatotoxicity in rats. Materials and Methods: The L. inermis leaves’ methanolic extract, which obtained by maceration, was orally administered in doses of 100 mg/kg and 200 mg/kg to the tested animals to assess its effects on serum levels of hepatotoxicity parameters, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), bilirubin, and total proteins along with histopathological liver sections examination, while silymarin (25 mg/kg), a potent hepatoprotective drug, was used as standard control. Results: The two doses of the plant extract showed dose-dependent hepatoprotective effect, as evident by the significant reduction (P < 0.05) in serum levels of AST, ALT, ALP, and bilirubin along with the improvement in histopathological liver sections compared to CCl4-only treated animals. Conclusion: As experimentally evident, it could be concluded that this plant material could provide a hepatoprotective effect that could be attributed to its antioxidant properties. PMID:27069719

  3. Safety and efficacy of hydroalcoholic extract from Lawsonia inermis leaves on lipid profile in alloxan-induced diabetic rats

    PubMed Central

    Singh, Surender; Verma, Nishikant; Karwasra, Ritu; Kalra, Prerna; Kumar, Rohit; Gupta, Yogendra Kumar

    2015-01-01

    Introduction: Dyslipidemia is one of the most common risk factor for cardiac-related disorders in diabetes mellitus. Diabetic dyslipidemia is characterized by hypertriglyceridemia, low high density lipoprotein and elevated low density lipoprotein concentration. Aim: To explore the effect of Lawsonia inermis hydroalcoholic extract (LIHE) for diabetic dyslipidemic activity along with its safety profile. Materials and Methods: LIHE administered at doses of 100, 200 and 400 mg/kg in rats after induction of hyperglycemia by alloxan. Insulin (1 IU/kg), glibenclamide (2.5 mg/kg), and metformin (100 mg/kg) were used as positive control and 1% gum acacia as normal control. Statistical analysis was performed using one-way analysis of variance, followed by Dunnett's t-test. Results: The percentage reduction in blood glucose level of LIHE at dose of 400 mg/kg was 39.08% on day 21 when compared to baseline (day 0), which is comparable to glibenclamide (44.77%) and metformin (46.30%). Decrease in blood glucose level exhibited significant improvement in lipid profile, plasma albumin, total plasma protein and serum creatinine. Conclusion: Results of this study demonstrated that LIHE significantly improved lipid and lipoprotein pattern observed in diabetic rats and this could be due to improvement in insulin secretion or action, thus has potential to be used in treatment of diabetes mellitus associated dyslipidemia. PMID:26730149

  4. Lousicidal activity of synthesized silver nanoparticles using Lawsonia inermis leaf aqueous extract against Pediculus humanus capitis and Bovicola ovis.

    PubMed

    Marimuthu, Sampath; Rahuman, Abdul Abdul; Santhoshkumar, Thirunavukkarasu; Jayaseelan, Chidambaram; Kirthi, Arivarasan Vishnu; Bagavan, Asokan; Kamaraj, Chinnaperumal; Elango, Gandhi; Zahir, Abdul Abduz; Rajakumar, Govindasamy; Velayutham, Kanayairam

    2012-11-01

    In the present work, we describe inexpensive, nontoxic, unreported and simple procedure for synthesis of silver nanoparticles (Ag NPs) using leaf aqueous extract of Lawsonia inermis as eco-friendly reducing and capping agent. The aim of the present study was to assess the lousicidal activity of synthesized Ag NPs against human head louse, Pediculus humanus capitis De Geer (Phthiraptera: Pediculidae), and sheep body louse, Bovicola ovis Schrank (Phthiraptera: Trichodectidae). Direct contact method was conducted to determine the potential of pediculocidal activity and impregnated method was used with slight modifications to improve practicality and efficiency of tested materials of synthesized Ag NPs against B. ovis. The synthesized Ag NPs characterized with the UV showing peak at 426 nm. X-ray diffraction (XRD) spectra clearly shows that the diffraction peaks in the pattern indexed as the silver with lattice constants. XRD analysis showed intense peaks at 2θ values of 38.34°, 44.59°, 65.04°, and 77.77° corresponding to (111), (200), (220), and (311) Bragg's reflection based on the fcc structure of Ag NPs. Fourier transform infrared spectroscopy (FTIR) spectra of Ag NPs exhibited prominent peaks at 3,422.13, 2,924.12, 2,851.76, 1,631.41, 1,381.60, 1,087.11, and 789.55 cm(-1). Scanning electron microscopy (SEM) micrograph showed mean size of 59.52 nm and aggregates of spherical shape Ag NPs. Energy dispersive X-ray spectroscopy (EDX) showed the complete chemical composition of the synthesized Ag NPs. In pediculocidal activity, the results showed that the optimal times for measuring percent mortality effects of synthesized Ag NPs were 26, 61, 84, and 100 at 5, 10, 15, and 20 min, respectively. The average percent mortality for synthesized Ag NPs was 33, 84, 91, and 100 at 10, 15, 20, and 35 min, respectively against B. ovis. The maximum activity was observed in the aqueous leaf extract of L. inermis, 1 mM AgNO(3) solution, and synthesized Ag NPs against P. humanus

  5. Antiproliferative and Apoptosis Inducing Effects of Non-Polar Fractions from Lawsonia inermis L. in Cervical (HeLa) Cancer Cells.

    PubMed

    Kumar, Manish; Kaur, Paramjeet; Kumar, Subodh; Kaur, Satwinderjeet

    2015-04-01

    Two non-polar fractions viz. hexane (Hex-LI) and chloroform fraction (CHCl3-LI) of Lawsonia inermis were studied for their antiproliferative potential in various cancer cell lines viz. HeLa, MCF-7, A549 and C6 glioma cells. Both the fractions showed more than 60 % of growth inhibition in all the tested cell lines at highest tested concentration. In clonogenic assay, different concentrations of Hex-LI and CHCl3-LI decreased the number and size of colonies as compared to control in HeLa cells. The apoptotic effects as nuclear condensation, fragmentation were visualized with Hoechst-33342 staining of HeLa cells using confocal microscope. Both fractions induced apoptotic cell death in human cervical carcinoma (HeLa) cells as evident from flow cytometric analysis carried out using Annexin V-FITC and propidium iodide dyes. CHCl3-LI treated cells significantly induced apoptosis (25.43 %) in comparison to control. Results from Neutral Comet assay demonstrated that both fractions induced double stranded breaks (DSB's) in HeLa cells. Our data indicated that Hex-LI and CHCl3-LI treated cells showed significant increase of 32.2 and 18.56 % reactive oxygen species (ROS) levels in DCFH-DA assay respectively. Further, experimental studies to decipher exact pathway via which these fractions induce cell death are in progress.

  6. Modulatory effect of henna leaf (Lawsonia inermis) on drug metabolising phase I and phase II enzymes, antioxidant enzymes, lipid peroxidation and chemically induced skin and forestomach papillomagenesis in mice.

    PubMed

    Dasgupta, Trisha; Rao, A R; Yadava, P K

    2003-03-01

    Henna leaf (Lawsonia inermis), commonly known as Mehndi is cultivated throughout India and is a very popular natural dye to color hand and hair. It is an integral part of indigenous culture, and is also known for its medicinal value. The effect of 200 and 400 mg/kg body weight of 80% ethanolic extract of the fresh leaves of Lawsonia inermis were examined on drug metabolizing phase-I and phase-II enzymes, antioxidant enzymes, glutathione content, lactate dehydrogenase and lipid peroxidation in the liver of 7 weeks old Swiss albino mice. Also anticarcinogenic potential of Henna leaf extract was studied adopting the protocol of benzo(a)pyrene induced forestomach and 7,12 dimethylbenz(a)anthracene (DMBA)-initiated and croton oil-promoted skin papillomagenesis. Our primary findings reveal the 'duel-acting' nature of henna leaf as deduced from its potential to induce only the phase-II enzyme activity, associated mainly with carcinogen detoxification in liver of mice and inhibit the phase I enzyme activities. The hepatic glutathione S-transferase and DT-diaphorase specific activities were elevated above basal (p < 0.005) level by Lawsonia inermis extract treatment. With reference to antioxidant enzymes the investigated doses were effective in increasing the hepatic glutathione reductase (GR), superoxide dismutase (SOD) and catalase activities significantly (from p < 0.05 to p < 0.005) at both the dose levels. Reduced glutathione (GSH) measured as non-protein sulphydryl was found to be significantly elevated in liver (p < 0.005) and in all the extrahepatic organs studied (from p < 0.05 to p < 0.005). Among the extrahepatic organs examined (forestomach, kidney and lung) glutathione S-transferase and DT-diaphorase level were increased in a dose independent manner (from p < 0.05 to p < 0.005). Chemopreventive response was measured by the average number of papillomas per mouse (tumor burden) as well as percentage of tumor bearing animals and tumor multiplicity. There was a

  7. Examining the anti-candidal activity of 10 selected Indian herbs and investigating the effect of Lawsonia inermis extract on germ tube formation, protease, phospholipase, and aspartate dehydrogenase enzyme activity in Candida albicans

    PubMed Central

    Ravichandran, Sripathy; Muthuraman, Sundararaman

    2016-01-01

    Objective: The objective of the study is to identify potential anti-candidal agents from natural resources and elucidate the effect of Lawsonia inermis extract on major virulent factors of Candida albicans. Materials and Methods: Plants, the most abundant and readily available resource of diverse bioactives, were chosen for the anti-candidal screening study. Ten different plants that were proven to have antimicrobial activity but not explored much for anti-candidal activity were chosen for this study. Ethyl acetate extract of these plant leaves were tested for the anti-candidal activity. Extracts with good anti-candidal activity were further screened for its effect in C. albicans germ tube formation and enzyme (protease, phospholipase, and aspartate dehydrogenase) activity. Results: Among 10 plants screened, L. inermis extract showed complete inhibition of C. albicans. On further evaluation, this extract completely inhibited C. albicans germ tube formation in serum until the end of incubation period (3 h). This extract also exhibited dose-dependent inhibitory activity against two major virulent enzymes of C. albicans, proteases (27–33%) and phospholipases (44.5%). In addition to it, this extract completely inhibited both the isoforms of constitutive candidal enzyme aspartate dehydrogenase, thereby affecting amino acid biosynthesis. Conclusion: Thus, this study confirms the anti-candidal potential of L. inermis and hence can be considered further for development of anti-candidal drug. PMID:26997722

  8. Seroprevalence of porcine proliferative enteropathy among wild boars in the Republic of Korea

    PubMed Central

    2014-01-01

    Background The importance of the wild boar as a reservoir of Lawsonia intracellularis was assessed by investigating the seroprevalence of this pathogen among wild boars in the Republic of Korea. The extent of exposure to L. intracellularis among wild boars (Sus scrofa coreanus) was monitored by a country-wide serological survey using an immunoperoxidase monolayer assay. Results In this study, antibodies to L. intracellularis were observed in 165 of 716 clinically healthy wild boars tested. The overall apparent prevalence calculated directly from the sample and the true prevalence calculated based on the accuracy of the test method were 23.0% (95% confidence interval: 20.0-26.3%) and 25.6% (95% confidence interval: 23.9-27.2%), respectively. Serologically positive animals were found in all the tested provinces. Conclusions Our results confirm that L. intracellularis is present in the wild boar population worldwide, even in Far East Asia. Despite the high seroprevalence shown in wild boars, further studies are warranted to evaluate their potential as a reservoir species because seroprevalence does not prove ongoing infection nor shedding of the bacteria in amounts sufficient to infect other animals. It should also be determined whether the wild boar, like the domestic pig, is a natural host of L. intracellularis. PMID:24393381

  9. The rabbit as an infection model for equine proliferative enteropathy

    PubMed Central

    Sampieri, Francesca; Allen, Andrew L.; Pusterla, Nicola; Vannucci, Fabio A.; Antonopoulos, Aphroditi J.; Ball, Katherine R.; Thompson, Julie; Dowling, Patricia M.; Hamilton, Don L.; Gebhart, Connie J.

    2013-01-01

    The objective of this study was to demonstrate the susceptibility of rabbits to Lawsonia intracellularis obtained from a case of clinical equine proliferative enteropathy (EPE). This is a preliminary step toward developing a rabbit infection model for studying pathogenesis and therapy of EPE in horses. Nine does were equally assigned to 3 groups. Animals in 2 groups (Group 1 and Group 2) were orally inoculated with different doses of cell-cultured L. intracellularis. Controls (Group 3) were sham-inoculated. Feces and blood were collected before the rabbits were infected and at 7, 14, and 21 days post-infection (DPI). Serum immunoglobulin G (IgG) titers were measured using an immunoperoxidase monolayer assay (IPMA) and fecal samples were analyzed with quantitative polymerase chain reaction (qPCR). A doe from each group was euthanized at 7, 14, and 21 DPI for collection and evaluation of intestinal samples. Tissues were stained by routine hematoxylin and eosin (H&E) method and immunohistochemistry (IHC) with L. intracellularis-specific mouse monoclonal antibody. At 14 DPI, serologic responses were detected in both infected groups, which maintained high titers through to 21 DPI. Lawsonia intracellularis DNA was detected in the feces of Group 2 on 7 DPI and in both infected groups on 14 DPI. Gross lesions were apparent in Group 1 and Group 2 on 14 DPI. Immunohistochemistry confirmed L. intracellularis antigen within cells of rabbits in Group 1 and Group 2 on 7, 14, and 21 DPI. No lesions, serologic response, shedding, or IHC labeling were found in Group 3 rabbits. This study describes an EPE rabbit model that simulates natural infection, as typical lesions, immune response, and fecal shedding were present. PMID:24082402

  10. The rabbit as an infection model for equine proliferative enteropathy.

    PubMed

    Sampieri, Francesca; Allen, Andrew L; Pusterla, Nicola; Vannucci, Fabio A; Antonopoulos, Aphroditi J; Ball, Katherine R; Thompson, Julie; Dowling, Patricia M; Hamilton, Don L; Gebhart, Connie J

    2013-04-01

    The objective of this study was to demonstrate the susceptibility of rabbits to Lawsonia intracellularis obtained from a case of clinical equine proliferative enteropathy (EPE). This is a preliminary step toward developing a rabbit infection model for studying pathogenesis and therapy of EPE in horses. Nine does were equally assigned to 3 groups. Animals in 2 groups (Group 1 and Group 2) were orally inoculated with different doses of cell-cultured L. intracellularis. Controls (Group 3) were sham-inoculated. Feces and blood were collected before the rabbits were infected and at 7, 14, and 21 days post-infection (DPI). Serum immunoglobulin G (IgG) titers were measured using an immunoperoxidase monolayer assay (IPMA) and fecal samples were analyzed with quantitative polymerase chain reaction (qPCR). A doe from each group was euthanized at 7, 14, and 21 DPI for collection and evaluation of intestinal samples. Tissues were stained by routine hematoxylin and eosin (H&E) method and immunohistochemistry (IHC) with L. intracellularis-specific mouse monoclonal antibody. At 14 DPI, serologic responses were detected in both infected groups, which maintained high titers through to 21 DPI. Lawsonia intracellularis DNA was detected in the feces of Group 2 on 7 DPI and in both infected groups on 14 DPI. Gross lesions were apparent in Group 1 and Group 2 on 14 DPI. Immunohistochemistry confirmed L. intracellularis antigen within cells of rabbits in Group 1 and Group 2 on 7, 14, and 21 DPI. No lesions, serologic response, shedding, or IHC labeling were found in Group 3 rabbits. This study describes an EPE rabbit model that simulates natural infection, as typical lesions, immune response, and fecal shedding were present.

  11. Emended descriptions of indole negative and indole positive isolates of Brachyspira (Serpulina) hyodysenteriae.

    PubMed

    Fellström, C; Karlsson, M; Pettersson, B; Zimmerman, U; Gunnarsson, A; Aspan, A

    1999-12-01

    Two type/reference strains of Brachyspira (B.) hyodysenteriae, 14 Belgian and German indole negative, and 14 Belgian, German and Swedish indole positive field isolates of strongly beta-haemolytic intestinal spirochaetes were compared by pulsed-field gel electrophoresis (PFGE) patterns, biochemical reaction patterns, 16S rDNA sequences and MIC determinations of six antibacterial substances. Three tests for indole production, including a spot indole test, were compared with congruent results. All field isolates were classified as B. hyodysenteriae due to a high genetic and phenotypic similarity with the type strains. The Belgian and German indole negative isolates had identical and unique PFGE patterns for the tested restriction enzymes MluI and SalI, as well as identical 16S rDNA sequences, and they could not be differentiated by any of the methods used. Seven unique PFGE patterns were achieved from the 14 indole positive field isolates. The patterns were identical and unique for epidemiologically related isolates. Type/reference strains and isolates without known relation to other tested isolates showed unique banding patterns. The MICs of tylosin, tiamulin, erythromycin, clindamycin, carbadox and virginiamycin were determined in broth for all isolates. In contrast to Belgian and German isolates, the majority of the Swedish field isolates were susceptible to tylosin, erythromycin and clindamycin. Probable pathways of infection for some of the Swedish isolates were determined. The PFGE patterns of epidemic clones of B. hyodysenteriae remained stable for a period of up to 8 years. In vivo development of resistance to macrolide and lincosamide antibiotics due to use of tylosin was clearly indicated for two epidemic clones. PMID:10596806

  12. Prevalence of swine viral and bacterial pathogens in rodents and stray cats captured around pig farms in Korea.

    PubMed

    Truong, Quang Lam; Seo, Tae Won; Yoon, Byung-Il; Kim, Hyeon-Cheol; Han, Jeong Hee; Hahn, Tae-Wook

    2013-12-30

    In 2008, 102 rodents and 24 stray cats from the areas around 9 pig farms in northeast South Korea were used to determine the prevalence of the following selected swine pathogens: ten viral pathogens [porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), rotavirus, classical swine fever virus (CSFV), porcine circovirus type 2 (PCV2), encephalomyocarditis virus (EMCV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine parvovirus (PPV), pseudorabies virus (PRV) and Japanese encephalitis virus (JEV)] and four bacterial pathogens (Brucella, Leptospira, Salmonella and Lawsonia intracellularis). In total, 1,260 tissue samples from 102 rodents and 24 stray cats were examined by specific PCR and RT-PCR assays, including tissue samples of the brain, tonsils, lungs, heart, liver, kidneys, spleen, small intestine, large intestine and mesenteric lymph nodes. The percentages of PCR-positive rodents for the porcine pathogens were as follows: 63.7% for Leptospira, 39.2% for Brucella, 6.8% for Salmonella, 15.7% for L. intracellularis, 14.7% for PCV2 and 3.9% for EMCV. The percentages of PCR-positive stray cats for the swine pathogens were as follows: 62.5% for Leptospira, 25% for Brucella, 12.5% for Salmonella, 12.5% for L. intracellularis and 4.2% for PEDV. These results may be helpful for developing control measures to prevent the spread of infectious diseases of pigs.

  13. Elevated fat skatole levels in immunocastrated, surgically castrated and entire male pigs with acute dysentery.

    PubMed

    Skrlep, Martin; Batorek, Nina; Bonneau, Michel; Fazarinc, Gregor; Segula, Blaž; Candek-Potokar, Marjeta

    2012-12-01

    Boar taint is due to androstenone and skatole (3-methyl-indole) accumulation in fat tissues. During a study to investigate the effect of immunocastration on fattening pigs, an outbreak of acute dysentery occurred caused by Lawsonia intracellularis and Brachyspira hyodysenteriae and resulted in cachexia and high mortality. Low androstenone levels in the immunocastrates (0.25 ± 0.04 μg/g liquid fat) suggested that the immunocastration had been effective, but unusually high skatole concentrations in fat tissues were found not only in entire males, but also in surgical castrates and immunocastrates (0.22 ± 0.15, 0.14 ± 0.08 and 0.18 ± 0.14 μg/g liquid fat, respectively). The findings suggest that boar taint can arise in cases of intestinal infections, even in castrated pigs.

  14. Detection of enteric pathogens in Turkey flocks affected with severe enteritis, in Brazil.

    PubMed

    Moura-Alvarez, Joelma; Nuñez, Luis F N; Astolfi-Ferreira, Claudete S; Knöbl, Terezinha; Chacón, Jorge L; Moreno, Andrea M; Jones, Richard C; Ferreira, Antonio J Piantino

    2014-08-01

    Twenty-two flocks of turkeys affected by enteric problems, with ages between 10 and 104 days and located in the Southern region of Brazil, were surveyed for turkey by PCR for turkey astrovirus type 2 (TAstV-2), turkey coronavirus (TCoV), hemorrhagic enteritis virus (HEV), rotavirus, reovirus, Salmonella spp., and Lawsonia intracellularis (Li) infections. Eleven profiles of pathogen combination were observed. The most frequently encountered pathogen combinations were TCoV-Li, followed by TCoV-TAstV-2-Li, TCoV-TastV-2. Only TCoV was detected as the sole pathogen in three flocks. Eight and 19 flocks of the 22 were positive for TAstV-2 and TCoV, respectively. Six were positive for Salmonella spp. and L. intracellularis was detected in 12 turkey flocks. Reovirus and HEV were not detected in this survey. These results throw new light on the multiple etiology of enteritis in turkeys. The implications of these findings and their correlation with the clinical signs are comprehensively discussed, illustrating the complexity of the enteric diseases.

  15. Detection of enteric pathogens in Turkey flocks affected with severe enteritis, in Brazil.

    PubMed

    Moura-Alvarez, Joelma; Nuñez, Luis F N; Astolfi-Ferreira, Claudete S; Knöbl, Terezinha; Chacón, Jorge L; Moreno, Andrea M; Jones, Richard C; Ferreira, Antonio J Piantino

    2014-08-01

    Twenty-two flocks of turkeys affected by enteric problems, with ages between 10 and 104 days and located in the Southern region of Brazil, were surveyed for turkey by PCR for turkey astrovirus type 2 (TAstV-2), turkey coronavirus (TCoV), hemorrhagic enteritis virus (HEV), rotavirus, reovirus, Salmonella spp., and Lawsonia intracellularis (Li) infections. Eleven profiles of pathogen combination were observed. The most frequently encountered pathogen combinations were TCoV-Li, followed by TCoV-TAstV-2-Li, TCoV-TastV-2. Only TCoV was detected as the sole pathogen in three flocks. Eight and 19 flocks of the 22 were positive for TAstV-2 and TCoV, respectively. Six were positive for Salmonella spp. and L. intracellularis was detected in 12 turkey flocks. Reovirus and HEV were not detected in this survey. These results throw new light on the multiple etiology of enteritis in turkeys. The implications of these findings and their correlation with the clinical signs are comprehensively discussed, illustrating the complexity of the enteric diseases. PMID:24817479

  16. Evaluating the risk of pathogen transmission from wild animals to domestic pigs in Australia.

    PubMed

    Pearson, Hayley E; Toribio, Jenny-Ann L M L; Lapidge, Steven J; Hernández-Jover, Marta

    2016-01-01

    Wild animals contribute to endemic infection in livestock as well as the introduction, reintroduction and maintenance of pathogens. The source of introduction of endemic diseases to a piggery is often unknown and the extent of wildlife contribution to such local spread is largely unexplored. The aim of the current study was to quantitatively assess the probability of domestic pigs being exposed to different pathogens from wild animals commonly found around commercial piggeries in Australia. Specifically, this study aims to quantify the probability of exposure to the pathogens Escherichia coli, Salmonella spp. and Campylobacter spp. from European starlings (Sturnus vulgarus); Brachyspira hyodysenteriae, Lawsonia intracellularis and Salmonella spp. from rats (Rattus rattus and Rattus norvegicus); and Mycoplasma hyopneumoniae, Leptospira spp., Brucella suis and L. intracellularis from feral pigs (Sus scrofa). Exposure assessments, using scenario trees and Monte Carlo stochastic simulation modelling, were conducted to identify potential pathways of introduction and calculate the probabilities of these pathways occurring. Input parameters were estimated from a national postal survey of commercial pork producers and from disease detection studies conducted for European starlings, rats and feral pigs in close proximity to commercial piggeries in Australia. Based on the results of the exposure assessments, rats presented the highest probability of exposure of pathogens to domestic pigs at any point in time, and L. intracellularis (median 0.13, 5% and 95%, 0.05-0.23) and B. hyodysenteriae (median 0.10, 0.05-0.19) were the most likely pathogens to be transmitted. Regarding European starlings, the median probability of exposure of domestic pigs to pathogenic E. coli at any point in time was estimated to be 0.03 (0.02-0.04). The highest probability of domestic pig exposure to feral pig pathogens at any point in time was found to be for M. hyopneumoniae (median 0.013, 0

  17. Evaluating the risk of pathogen transmission from wild animals to domestic pigs in Australia.

    PubMed

    Pearson, Hayley E; Toribio, Jenny-Ann L M L; Lapidge, Steven J; Hernández-Jover, Marta

    2016-01-01

    Wild animals contribute to endemic infection in livestock as well as the introduction, reintroduction and maintenance of pathogens. The source of introduction of endemic diseases to a piggery is often unknown and the extent of wildlife contribution to such local spread is largely unexplored. The aim of the current study was to quantitatively assess the probability of domestic pigs being exposed to different pathogens from wild animals commonly found around commercial piggeries in Australia. Specifically, this study aims to quantify the probability of exposure to the pathogens Escherichia coli, Salmonella spp. and Campylobacter spp. from European starlings (Sturnus vulgarus); Brachyspira hyodysenteriae, Lawsonia intracellularis and Salmonella spp. from rats (Rattus rattus and Rattus norvegicus); and Mycoplasma hyopneumoniae, Leptospira spp., Brucella suis and L. intracellularis from feral pigs (Sus scrofa). Exposure assessments, using scenario trees and Monte Carlo stochastic simulation modelling, were conducted to identify potential pathways of introduction and calculate the probabilities of these pathways occurring. Input parameters were estimated from a national postal survey of commercial pork producers and from disease detection studies conducted for European starlings, rats and feral pigs in close proximity to commercial piggeries in Australia. Based on the results of the exposure assessments, rats presented the highest probability of exposure of pathogens to domestic pigs at any point in time, and L. intracellularis (median 0.13, 5% and 95%, 0.05-0.23) and B. hyodysenteriae (median 0.10, 0.05-0.19) were the most likely pathogens to be transmitted. Regarding European starlings, the median probability of exposure of domestic pigs to pathogenic E. coli at any point in time was estimated to be 0.03 (0.02-0.04). The highest probability of domestic pig exposure to feral pig pathogens at any point in time was found to be for M. hyopneumoniae (median 0.013, 0

  18. Prevalence of Campylobacter spp. relative to other enteric pathogens in grow-finish pigs with diarrhea.

    PubMed

    Burrough, Eric; Terhorst, Samantha; Sahin, Orhan; Zhang, Qijing

    2013-08-01

    Salmonella spp., Lawsonia intracellularis, and Brachyspira spp. are pathogens commonly associated with diarrhea in growing and finishing pigs. Brachyspira spp. infection has recently reemerged as a significant concern due to an increase in the incidence of swine dysentery; however, the mechanisms underlying this increase in dysentery remain largely unknown. Pigs are also well-recognized as potential carriers of Campylobacter spp., particularly Campylobacter coli, yet enteric disease in swine associated with infection by these bacteria is considered uncommon and diagnosis has historically been based upon exclusion of other causes. Accordingly, Campylobacter culture is often excluded in routine diagnostic testing of cases of porcine enterocolitis and the incidence of infection is therefore largely unknown. In this study, feces from 155 cases of clinical diarrhea in grow-finish pigs submitted to the Iowa State University Veterinary Diagnostic Laboratory were cultured for Campylobacter spp. in addition to other testing as indicated for routine diagnostic investigation. Campylobacter culture was positive from 82.6% (128/155) of samples with C. coli accounting for 75% of isolates and Campylobacter jejuni for the remaining 25%. In 14.8% (23/155) of cases a Campylobacter spp. was the sole infectious agent detected; however, there was no association with any particular Campylobacter spp. Interestingly, for those cases with a laboratory diagnosis of Brachyspira-associated disease, 100% (15/15) were also culture positive for Campylobacter spp. suggesting a possible interrelationship between these bacteria in the pig gut. No association was noted between Campylobacter culture results and infection with either Salmonella spp. or L. intracellularis. PMID:23792232

  19. Dyeing behaviour of gamma irradiated cotton fabric using Lawson dye extracted from henna leaves (Lawsonia inermis)

    NASA Astrophysics Data System (ADS)

    Rehman, Fazal-ur; Adeel, Shahid; Qaiser, Summia; Ahmad Bhatti, Ijaz; Shahid, Muhammad; Zuber, Mohammad

    2012-11-01

    Dyeing behavior of gamma irradiated cotton fabric using Lawson dye extracted from henna leaves has been investigated. Cotton and dye powder are irradiated to different absorbed doses of 2, 4, 6, 8 and 10 kGy using Cs-137 gamma irradiator. The dyeing parameters such as dyeing time, electrolyte (salt) concentration and mordant concentrations using copper and iron as mordants are optimized. Dyeing is performed using un-irradiated and irradiated cotton with dye solutions and their color strength values are evaluated in CIE Lab system using Spectraflash -SF650. Methods suggested by International Standard Organization (ISO) have been employed to investigate the colourfastness properties such as colourfastness to light, washing and rubbing of irradiated dyed fabric. It is found that gamma ray treatment of cotton dyed with extracts of henna leaves has significantly improved the color strength as well as enhanced the rating of fastness properties.

  20. Comparative Tissue Stainability of Lawsonia inermis (Henna) and Eosin as Counterstains to Hematoxylin in Brain Tissues.

    PubMed

    Alawa, Judith N; Gideon, Gbenga O; Adetiba, Bamidele; Alawa, Clement B

    2015-04-01

    We hyposthesized that henna staining could provide an alternative to eosin when used as a counterstain to hematoxylin for understanding basic neurohistological principles. Therefore, this study was aimed at investigating the suitability of henna as counterstain to hematoxylin for the demonstration of the layer stratification and cellular distribution in the brain tissue. Henna stained nervous tissue by reacting with the basic elements in proteins via its amino groups. It stained the neuropil and connective tissue membranes brown and effectively outlined the perikarya of neurons with no visible nuclei demonstrating that it is an acidic dye. Henna as a counterstain to hematoxylin demonstrated reliability as a new neurohistological stain. It facilitated identification of cortical layer stratification and cellular distribution in brain tissue sections from Wistar rats. This was comparable to standard hematoxylin and eosin staining as morphological and morphometrical analyses of stained cells did not show significant differences in size or number. This study presents a method for staining with henna and demonstrates that although henna and eosin belong to different dye groups (anthraquinone and xanthenes, respectively) based on their chromophores, they share similar staining techniques and thus could be used interchangeably in neurohistology.

  1. The European ban on growth-promoting antibiotics and emerging consequences for human and animal health.

    PubMed

    Casewell, Mark; Friis, Christian; Marco, Enric; McMullin, Paul; Phillips, Ian

    2003-08-01

    Following the ban of all food animal growth-promoting antibiotics by Sweden in 1986, the European Union banned avoparcin in 1997 and bacitracin, spiramycin, tylosin and virginiamycin in 1999. Three years later, the only attributable effect in humans has been a diminution in acquired resistance in enterococci isolated from human faecal carriers. There has been an increase in human infection from vancomycin-resistant enterococci in Europe, probably related to the increased in usage of vancomycin for the treatment of methicillin-resistant staphylococci. The ban of growth promoters has, however, revealed that these agents had important prophylactic activity and their withdrawal is now associated with a deterioration in animal health, including increased diarrhoea, weight loss and mortality due to Escherichia coli and Lawsonia intracellularis in early post-weaning pigs, and clostridial necrotic enteritis in broilers. A directly attributable effect of these infections is the increase in usage of therapeutic antibiotics in food animals, including that of tetracycline, aminoglycosides, trimethoprim/sulphonamide, macrolides and lincosamides, all of which are of direct importance in human medicine. The theoretical and political benefit of the widespread ban of growth promoters needs to be more carefully weighed against the increasingly apparent adverse consequences. PMID:12837737

  2. Distribution and chemical coding of intramural neurons in the porcine ileum during proliferative enteropathy.

    PubMed

    Pidsudko, Z; Kaleczyc, J; Wasowicz, K; Sienkiewicz, W; Majewski, M; Zajac, W; Lakomy, M

    2008-01-01

    Enteric neurons are highly adaptive in their response to various pathological processes including inflammation, so the aim of this study was to describe the chemical coding of neurons in the ileal intramural ganglia in porcine proliferative enteropathy (PPE). Accordingly, juvenile Large White Polish pigs with clinically diagnosed Lawsonia intracellularis infection (PPE; n=3) and a group of uninfected controls (C; n=3) were studied. Ileal tissue from each animal was processed for dual-labelling immunofluorescence using antiserum specific for protein gene product 9.5 (PGP 9.5) in combination with antiserum to one of: vasoactive intestinal polypeptide (VIP), substance P (SP), calcitonin gene-related peptide (CGRP), somatostatin (SOM), neuropeptide Y (NPY) or galanin (GAL). In infected pigs, enteric neurons were found in ganglia located within three intramural plexuses: inner submucosal (ISP), outer submucosal (OSP) and myenteric (MP). Immunofluorescence labelling revealed increases in the number of neurons containing GAL, SOM, VIP and CGRP in pigs with PPE. Neuropeptides may therefore have an important role in the function of porcine enteric local nerve circuits under pathological conditions, when the nervous system is stressed, challenged or afflicted by disease such as PPE. However, further studies are required to determine the exact physiological relevance of the observed adaptive changes. PMID:18061202

  3. Disease risks associated with free-ranging wild boar in Saskatchewan.

    PubMed

    McGregor, Glenna F; Gottschalk, Marcelo; Godson, Dale L; Wilkins, Wendy; Bollinger, Trent K

    2015-08-01

    This study investigated the disease status of Saskatchewan's feral wild boar population. Whole carcasses, tissue samples, and/or serum from 81 hunter-killed boars from Saskatchewan were submitted to the Canadian Wildlife Health Cooperative (CWHC) between 2009 and 2014. Serological tests were negative for PRRS, H1N1, and H3N2 swine influenza, PCV-2, and TGE/PRCV in 22/22 boars and for Toxoplasma gondii and Mycoplasma hyopneumoniae in 20/20 boars. Of 20 boars whose sera were tested 20 were positive for Actinobacillus pleuropneumoniae, with 7 positive for, among other strains, serotype 14; 16 were positive for Lawsonia intracellularis, 1 was positive and 6 were suspicious for Salmonella spp. Polymerase chain reaction tests were negative for PRRS and PCV2 in 58/58 boars and positive for Torque teno virus in 1/8 boars. Digestion assays were negative for Trichinella spp. in 22/22 boars. The high seroprevalence of A. pleuropneumoniae serotype 14 is noteworthy as this serotype has not been previously reported in North America.

  4. 21 CFR 558.600 - Tiamulin.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... Sponsor (i) (ii) 35 1. For control of swine dysentery associated with Brachyspira (formerly Serpulina or... history of swine dysentery but where signs of disease have not yet occurred or following approved... chlortetracycline, and control of swine dysentery associated with Brachyspira (formerly Serpulina or...

  5. 21 CFR 558.600 - Tiamulin.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... Sponsor (i) (ii) 35 1. For control of swine dysentery associated with Brachyspira (formerly Serpulina or... history of swine dysentery but where signs of disease have not yet occurred or following approved... chlortetracycline, and control of swine dysentery associated with Brachyspira (formerly Serpulina or...

  6. Lead exposure from the use of Lawsonia inermis (henna) in temporary paint-on-tattooing and hair dying.

    PubMed

    Jallad, Karim N; Espada-Jallad, Cyntia

    2008-07-01

    This study reports the evaluation of a number of spectroscopic techniques used in identifying and quantifying the presence of lead in twelve commercial and traditional henna samples. The lead levels found in henna were low with concentrations ranging from 2.29 ppm to 65.98 ppm. Henna is used as a traditional cosmetic and remedy in the Middle East, Far East, and North Africa. The very low concentrations of lead measured in these henna samples were reassuring; however, the cumulative effects of prolonged lead exposure may be of concern. Thus, the use of henna especially among children may constitute a public health risk. PMID:18407319

  7. 21 CFR 558.600 - Tiamulin.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... dysentery associated with Brachyspira (formerly Serpulina or Treponema) hyodysenteriae susceptible to tiamulin Feed continuously as sole ration on premises with a history of swine dysentery but where signs of... Pasteurella multocida susceptible to chlortetracycline, and control of swine dysentery associated...

  8. 21 CFR 558.600 - Tiamulin.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... dysentery associated with Brachyspira (formerly Serpulina or Treponema) hyodysenteriae susceptible to tiamulin Feed continuously as sole ration on premises with a history of swine dysentery but where signs of... Pasteurella multocida susceptible to chlortetracycline, and control of swine dysentery associated...

  9. Intrinsic water use efficiency controls the adaptation to high salinity in a semi-arid adapted plant, henna (Lawsonia inermis L.).

    PubMed

    Fernández-García, Nieves; Olmos, Enrique; Bardisi, Enas; García-De la Garma, Jesús; López-Berenguer, Carmen; Rubio-Asensio, José Salvador

    2014-03-01

    Adaptation to salinity of a semi-arid inhabitant plant, henna, is studied. The salt tolerance mechanisms are evaluated in the belief that gas exchange (water vapor and CO2) should play a key role on its adaptation to salt stress because of the strong evaporation conditions and soil water deficit in its natural area of distribution. We grow henna plants hydroponically under controlled climate conditions and expose them to control (0mM NaCl), and two levels of salinity; medium (75mM NaCl) and high (150mM NaCl). Relative growth rate (RGR), biomass production, whole plant and leaf structure and ultrastructure adaptation, gas exchange, chlorophyll fluorescence, nutrients location in leaf tissue and its balance in the plant are studied. RGR and total biomass decreased as NaCl concentration increased in the nutrient solution. At 75mM NaCl root biomass was not affected by salinity and RGR reached similar values to control plants at the end of the experiment. At this salinity level henna plant responded to salinity decreasing shoot to root ratio, increasing leaf specific mass (LSM) and intrinsic water use efficiency (iWUE), and accumulating high concentrations of Na(+) and Cl(-) in leaves and root. At 150mM NaCl growth was severely reduced but plants reached the reproductive phase. At this salinity level, no further decrease in shoot to root ratio or increase in LSM was observed, but plants increased iWUE, maintaining water status and leaf and root Na(+) and Cl(-) concentrations were lower than expected. Moreover, plants at 150mM NaCl reallocated carbon to the root at the expense of the shoot. The effective PSII quantum yield [Y(II)] and the quantum yield of non-regulated energy dissipation [Y(NO)] were recovered over time of exposure to salinity. Overall, iWUE seems to be determinant in the adaptation of henna plant to high salinity level, when morphological adaptation fails. PMID:24484959

  10. Effect of feeding sodium butyrate in the late finishing period on Salmonella carriage, seroprevalence, and growth of finishing pigs.

    PubMed

    Walia, Kavita; Argüello, Hector; Lynch, Helen; Leonard, Finola C; Grant, Jim; Yearsley, Dermot; Kelly, Sinead; Duffy, Geraldine; Gardiner, Gillian E; Lawlor, Peadar G

    2016-09-01

    Pork is an important source of human salmonellosis and low-cost on-farm control measures may provide a useful element in reducing the prevalence of this pathogen in food. This study investigated the effectiveness of dietary supplementation with sodium butyrate administered to finisher pigs for ∼4-weeks prior to slaughter to control Salmonella shedding on highly contaminated farms. Two trials (A and B) were conducted on two commercial pig farms, which had a history of high Salmonella seroprevalence. In both trials, pens (14 pens of 12 pigs/pen in Trial A and 12 pens of 12-17 pigs/pen in Trial B) were randomly assigned to a control (finisher feed without additive) or a treatment group (the same feed with 3kg sodium butyrate/t) for 24-28days, depending on the trial. Faeces were collected from each pig on days 0, 12 and 24/28, and blood, caecal digesta and ileocaecal/mesenteric lymph nodes were collected from the slaughterhouse. Pigs were weighed at the start and end of the trials, feed intake was recorded, and carcass quality parameters were recorded at slaughter. In Trial A, Salmonella shedding was reduced in the treatment compared to the control group at the end of the trial (30% versus 57% probability of detecting Salmonella in faeces, respectively; p<0.001). This reflected the serology results, with detection of a lower seroprevalence in the treatment compared to the control group using the 20% optical density cut-off (69.5% versus 89%; p=0.001). However, no effect on faecal shedding or seroprevalance was observed in Trial B, which may be explained by the detection of a concomitant infection with Lawsonia intracellularis. No significant differences in Salmonella recovery rates were observed in the caecal digesta or lymph nodes in either trial. Furthermore, feed intake, weight gain, and feed conversion efficiency (FCE) did not differ between groups (p>0.05) in either trial. Numerical improvements in weight gain and FCE were found with sodium butyrate treatment

  11. Effect of feeding sodium butyrate in the late finishing period on Salmonella carriage, seroprevalence, and growth of finishing pigs.

    PubMed

    Walia, Kavita; Argüello, Hector; Lynch, Helen; Leonard, Finola C; Grant, Jim; Yearsley, Dermot; Kelly, Sinead; Duffy, Geraldine; Gardiner, Gillian E; Lawlor, Peadar G

    2016-09-01

    Pork is an important source of human salmonellosis and low-cost on-farm control measures may provide a useful element in reducing the prevalence of this pathogen in food. This study investigated the effectiveness of dietary supplementation with sodium butyrate administered to finisher pigs for ∼4-weeks prior to slaughter to control Salmonella shedding on highly contaminated farms. Two trials (A and B) were conducted on two commercial pig farms, which had a history of high Salmonella seroprevalence. In both trials, pens (14 pens of 12 pigs/pen in Trial A and 12 pens of 12-17 pigs/pen in Trial B) were randomly assigned to a control (finisher feed without additive) or a treatment group (the same feed with 3kg sodium butyrate/t) for 24-28days, depending on the trial. Faeces were collected from each pig on days 0, 12 and 24/28, and blood, caecal digesta and ileocaecal/mesenteric lymph nodes were collected from the slaughterhouse. Pigs were weighed at the start and end of the trials, feed intake was recorded, and carcass quality parameters were recorded at slaughter. In Trial A, Salmonella shedding was reduced in the treatment compared to the control group at the end of the trial (30% versus 57% probability of detecting Salmonella in faeces, respectively; p<0.001). This reflected the serology results, with detection of a lower seroprevalence in the treatment compared to the control group using the 20% optical density cut-off (69.5% versus 89%; p=0.001). However, no effect on faecal shedding or seroprevalance was observed in Trial B, which may be explained by the detection of a concomitant infection with Lawsonia intracellularis. No significant differences in Salmonella recovery rates were observed in the caecal digesta or lymph nodes in either trial. Furthermore, feed intake, weight gain, and feed conversion efficiency (FCE) did not differ between groups (p>0.05) in either trial. Numerical improvements in weight gain and FCE were found with sodium butyrate treatment

  12. Corrigendum to "Natural dye extract of lawsonia inermis seed as photo sensitizer for titanium dioxide based dye sensitized solar cells" [Spectrochim. Acta A Mol. Biomol. Spectrosc. 128 (2014) 420-426

    NASA Astrophysics Data System (ADS)

    Ananth, S.; Vivek, P.; Arumanayagam, T.; Murugakoothan, P.

    2015-05-01

    The authors regret to inform that the following text in page No. 425, "The conventionally prepared TiO2 based DSSC exhibits the observed values of Isc = 1.6 mA cm-2, Voc = 0.48 V and the calculated value of FF is 64.32%. Similarly, for pre dye treated TiO2 based DSSC, the observed values of Isc = 2.99 mA cm-2, Voc = 0.50 V and the calculated value of FF is 66.9%" should be read as, "The conventionally prepared TiO2 based DSSC exhibits the observed values of Isc = 2.99 mA cm-2, Voc = 0.50 V and the calculated value of FF is 66.9%. Similarly, for pre dye treated TiO2 based DSSC, the observed values of Isc = 4.2 mA cm-2, Voc = 0.56 V and the calculated value of FF is 62.5%". Though the text is typed wrongly, the I-V graph, the discussions and our conclusions are as same as that of our published article.

  13. PHYSICO-CHEMICAL PROFILE OF SOME COLOURING PLANTS USED IN HOMOEOPATHY

    PubMed Central

    Subramaniam, P.; Sunilkumar

    2001-01-01

    The objective of this paper deal with the physico chemical aspects of certain colouring plants namely. Bixa orellana Linn. (Leaves) and Lawsonia inermis Linn (Leaves). The determined data under the physico chemical, chromatographic and spectrophotometric studies can be taken as a pharmacopoeial standards. PMID:22557040

  14. Physico-chemical profile of some colouring plants used in homoeopathy.

    PubMed

    Subramaniam, P; Sunilkumar

    2001-10-01

    The objective of this paper deal with the physico chemical aspects of certain colouring plants namely. Bixa orellana Linn. (Leaves) and Lawsonia inermis Linn (Leaves). The determined data under the physico chemical, chromatographic and spectrophotometric studies can be taken as a pharmacopoeial standards. PMID:22557040

  15. Complete genome sequence of Brachyspira murdochii type strain (56-150T)

    SciTech Connect

    Pati, Amrita; Sikorski, Johannes; Gronow, Sabine; Munk, Christine; Lapidus, Alla L.; Copeland, A; Glavina Del Rio, Tijana; Nolan, Matt; Lucas, Susan; Chen, Feng; Tice, Hope; Cheng, Jan-Fang; Han, Cliff; Detter, J. Chris; Bruce, David; Tapia, Roxanne; Goodwin, Lynne A.; Pitluck, Sam; Liolios, Konstantinos; Ivanova, N; Mavromatis, K; Mikhailova, Natalia; Chen, Amy; Palaniappan, Krishna; Land, Miriam L; Hauser, Loren John; Chang, Yun-Juan; Jeffries, Cynthia; Spring, Stefan; Rohde, Manfred; Goker, Markus; Bristow, James; Eisen, Jonathan; Markowitz, Victor; Hugenholtz, Philip; Klenk, Hans-Peter; Kyrpides, Nikos C

    2010-01-01

    Brachyspira murdochii Stanton et al. 1992 is a non-pathogenic but host-associated spirochete of the family Brachyspiraceae. Initially isolated from the intestinal content of a healthy swine, the group B spirochaetes were first described under the basonym Serpulina murdochii. Members of the family Brachyspiraceae are of great phylogenetic interest because of the extremely isolated location of this family within the phylum Spirochaetes . Here we describe the features of this organism, together with the complete genome sequence and annotation. This is the first completed genome sequence of a type strain of a member of the family Brachyspiraceaeand only the second genome sequence from a member of the genus Brachyspira. The 3,241,804 bp long genome with its 2,893 protein-coding and 40 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project.

  16. Cloning and DNA Sequence Analysis of an Immunogenic Glucose-Galactose MglB Lipoprotein Homologue from Brachyspira pilosicoli, the Agent of Colonic Spirochetosis†

    PubMed Central

    Zhang, P.; Cheng, X.; Duhamel, G. E.

    2000-01-01

    Colonic spirochetosis (CS) is a newly emerging infectious disease of humans and animals caused by the pathogenic spirochete Brachyspira (formerly Serpulina) pilosicoli. The purpose of this study was to characterize an antigen that was recognized by antibodies present in sera of challenge-exposed pigs. The gene encoding the antigen was identified by screening a plasmid library of human B. pilosicoli strain SP16 (ATCC 49776) genomic DNA with hyperimmune and convalescent swine sera. The predicted amino acid sequence encoded by the cloned B. pilosicoli gene had a high degree of similarity and identity to glucose-galactose MglB lipoprotein. Located 106 bp downstream of the putative mglB gene was a 3′-truncated open reading frame with 73.8% similarity and 66.3% identity to mglA of Escherichia coli, suggesting a gene arrangement within an operon which is similar to those of other bacteria. A single copy of the gene was present in B. pilosicoli, and homologous sequences were widely conserved among porcine intestinal spirochetes Serpulina intermedia, Brachyspira innocens, Brachyspira murdochii, and the avian Brachyspira alvinipulli, but not in porcine Brachyspira hyodysenteriae, human Brachyspira aalborgi, and porcine Treponema succinifaciens. The deduced molecular weight of the mature MglB lipoprotein was consistent with expression by the cloned gene of a polypeptide with an apparent molecular weight of 36,000, as determined by Western blot analysis and [3H]palmitate labeling. Because mucin is the principal constituent of the colonic mucus gel and consists of glycoproteins that can serve as the substrate for growth and chemotaxis of B. pilosicoli in vitro, a role for MglB in mucosal localization of the spirochete appears consistent with the pathogenesis of CS. However, the presence of homologous sequences in closely related but nonpathogenic commensal spirochetes suggests that other virulence determinants may be required for pathogenesis. PMID:10899855

  17. Review on medicinal uses, pharmacological, phytochemistry and immunomodulatory activity of plants.

    PubMed

    Akram, M; Hamid, A; Khalil, A; Ghaffar, A; Tayyaba, N; Saeed, A; Ali, M; Naveed, A

    2014-01-01

    Since ancient times, plants have been an exemplary source of medicine. Researchers have discovered some important compounds from plants. The present work constitutes a review of the medicinal plants whose immunomodulant activity has been proven. We performed PUBMED, EMBASE, Google scholar searches for research papers of medicinal plants having immunomodulant activity. Medicinal plants used by traditional physicians or reported as having immunomodulant activity include Acacia concocinna, Camellia sinensis, Lawsonia inermis Linn, Piper longum Linn, Gelidium amansii, Petroselinum crispum, Plantago major and Allium sativum. Immunomodulant activities of some of these medicinal plants have been investigated. The medicinal plants documented have immunomodulant activity and should be further investigated via clinical trial. PMID:25280022

  18. Intra-amoeba multiplication induces chemotaxis and biofilm colonization and formation for Legionella.

    PubMed

    Bigot, Renaud; Bertaux, Joanne; Frere, Jacques; Berjeaud, Jean-Marc

    2013-01-01

    Legionella pneumophila, a facultative intracellular bacterium, is the causative agent of legionellosis. In the environment this pathogenic bacterium colonizes the biofilms as well as amoebae, which provide a rich environment for the replication of Legionella. When seeded on pre-formed biofilms, L. pneumophila was able to establish and survive and was only found at the surface of the biofilms. Different phenotypes were observed when the L. pneumophila, used to implement pre-formed biofilms or to form mono-species biofilms, were cultivated in a laboratory culture broth or had grown intracellulary within the amoeba. Indeed, the bacteria, which developed within the amoeba, formed clusters when deposited on a solid surface. Moreover, our results demonstrate that multiplication inside the amoeba increased the capacity of L. pneumophila to produce polysaccharides and therefore enhanced its capacity to establish biofilms. Finally, it was shown that the clusters formed by L. pneumophila were probably related to the secretion of a chemotaxis molecular agent.

  19. The morphology of the Legionnaires' disease organism.

    PubMed Central

    Katz, S. M.

    1978-01-01

    The Pennsylvania Department of Health, Bureau of Laboratories, has isolated the Legionnaires disease organism from two patients with Legionnaires' disease proved by serologic techniques. We have studied the morphology which the isolate assumes in yolk sac tissue and on bacteriologic media. The organism was Giménez-positive and gram-variable. Using an indirect immunofluorescent procedure, it was shown to react with convalescent serum samples taken from patients who had Legionnaires' disease. The organism multiplies by binary fission extracellularly and intracellulary; is both coccoid and bacillary in form; and contains characteristic cytoplasm, nucleoids, a cytoplasmic membrane, and a small cell wall of variable size. It may produce spores of unusual appearance. Intracellular replication characteristically occurs within vacuoles. The Legionnaires' disease organism conforms to the morphologic criteria for a prokaryocyte. Images Figure 10 Figure 6 Figure 2 Figure 11 Figure 3 Figure 7 Figure 8 Figure 4 Figure 9 Figure 1 Figure 5 PMID:629329

  20. Intra-Amoeba Multiplication Induces Chemotaxis and Biofilm Colonization and Formation for Legionella

    PubMed Central

    Bigot, Renaud; Bertaux, Joanne; Frere, Jacques; Berjeaud, Jean-Marc

    2013-01-01

    Legionella pneumophila, a facultative intracellular bacterium, is the causative agent of legionellosis. In the environment this pathogenic bacterium colonizes the biofilms as well as amoebae, which provide a rich environment for the replication of Legionella. When seeded on pre-formed biofilms, L. pneumophila was able to establish and survive and was only found at the surface of the biofilms. Different phenotypes were observed when the L. pneumophila, used to implement pre-formed biofilms or to form mono-species biofilms, were cultivated in a laboratory culture broth or had grown intracellulary within the amoeba. Indeed, the bacteria, which developed within the amoeba, formed clusters when deposited on a solid surface. Moreover, our results demonstrate that multiplication inside the amoeba increased the capacity of L. pneumophila to produce polysaccharides and therefore enhanced its capacity to establish biofilms. Finally, it was shown that the clusters formed by L. pneumophila were probably related to the secretion of a chemotaxis molecular agent. PMID:24205008

  1. The morphology of the Legionnaires' disease organism.

    PubMed

    Katz, S M

    1978-03-01

    The Pennsylvania Department of Health, Bureau of Laboratories, has isolated the Legionnaires disease organism from two patients with Legionnaires' disease proved by serologic techniques. We have studied the morphology which the isolate assumes in yolk sac tissue and on bacteriologic media. The organism was Giménez-positive and gram-variable. Using an indirect immunofluorescent procedure, it was shown to react with convalescent serum samples taken from patients who had Legionnaires' disease. The organism multiplies by binary fission extracellularly and intracellulary; is both coccoid and bacillary in form; and contains characteristic cytoplasm, nucleoids, a cytoplasmic membrane, and a small cell wall of variable size. It may produce spores of unusual appearance. Intracellular replication characteristically occurs within vacuoles. The Legionnaires' disease organism conforms to the morphologic criteria for a prokaryocyte. PMID:629329

  2. Acute renal failure and intravascular hemolysis following henna ingestion.

    PubMed

    Qurashi, Hala E A; Qumqumji, Abbas A A; Zacharia, Yasir

    2013-05-01

    The powder of henna plant (Lawsonia inermis Linn.) is extensively used as a decorative skin paint for nail coloring and as a hair dye. Most reports of henna toxicity have been attributed to adding a synthetic dye para-phenylenediamine (PPD). PPD is marketed as black henna added to natural henna to accentuate the dark color and shorten the application time. PPD toxicity is well known and extensively reported in medical literature. We report a case of a young Saudi male who presented with characteristic features of acute renal failure and intravascular hemolysis following ingestion of henna mixture. Management of PPD poisoning is only supportive and helpful only if instituted early. Diagnosis requires a high degree of clinical suspicion, as the clinical features are quite distinctive. PMID:23640630

  3. Anna Pavala Sindhooram--an antiatherosclerotic Indian drug.

    PubMed

    Shanmugasundaram, K R; Seethapathy, P G; Shanmugasundaram, E R

    1983-05-01

    The procedure for the preparation of Anna Pavala Sindhooram (APS), a drug based on the concepts of Indian medicine for the prevention and reversal of the atherosclerotic disease process is described in detail. The uniformity in the composition of samples of the drug obtained in 10 batches, prepared over a period of 5 years is evident from the chemical analysis of the mineral constituents. The possibility of iodine, copper, iron, calcium and magnesium present in the Anna Pavala Sindhooram, acting metabolically to reduce hypercholesterolemia is discussed. The ingredients used are green vitriol (Annabedi or ferrous sulphate), coral reef (Corallium rubrum or Pavalam), leaves of Acalypha indica (Kuppaimeni), Lippia nodiflora (Poduthalai), Vinca rosea (Nityakalyani), Lawsonia alba (maruthondri) and Cynodon dactylon (Arugampul) and the flowers of Hibiscus rosasinensis (Chemparathampoo) and the ripe fruits of Phyllanthus emblica (nellikkai). Sindhooram is the chief therapeutic form of herbo-mineral preparations used in the Sidha system of Indian medicine. PMID:6876850

  4. Occurrence of pathogens in wild rodents caught on Swedish pig and chicken farms.

    PubMed

    Backhans, A; Jacobson, M; Hansson, I; Lebbad, M; Lambertz, S Thisted; Gammelgård, E; Saager, M; Akande, O; Fellström, C

    2013-09-01

    A total of 207 wild rodents were caught on nine pig farms, five chicken farms and five non-farm locations in Sweden and surveyed for a selection of bacteria, parasites and viruses. Lawsonia intracellularia and pathogenic Yersinia enterocolitica were only detected in rodents on pig farms (9% and 8% prevalence, respectively) which indicate that these agents are more likely to be transmitted to rodents from pigs or the environment on infected farms. Brachyspira hyodysenteriae (1%), Brachyspira intermedia (2%), Campylobacter jejuni (4%), Campylobacter upsaliensis (2%), leptospires (7%) and encephalomyocarditis virus (9%) were also detected from rodents not in contact with farm animals. Giardia and Cryptosporidium spp. were common, although no zoonotic types were verified, and Salmonella enterica was isolated from 1/11 mice on one farm but not detected by PCR from any of the rodents. Trichinella spp. and Toxoplasma gondii were not detected. PMID:23174339

  5. Effect of certain bioactive plant extracts on clinical isolates of beta-lactamase producing methicillin resistant Staphylococcus aureus.

    PubMed

    Aqil, Farrukh; Khan, M Sajjad A; Owais, Mohd; Ahmad, Iqbal

    2005-01-01

    Ethanolic extracts and some fractions from 10 Indian medicinal plants, known for antibacterial activity, were investigated for their ability to inhibit clinical isolates of beta-lactamase producing methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive S. aureus (MSSA). Synergistic interaction of plant extracts with certain antibiotics was also evaluated. The MRSA test strains were found to be multi-drug resistant and also exhibited high level of resistance to common beta-lactam antibiotics. These strains produced beta-lactamases, which hydrolyze one or other beta-lactam antibiotics, tested. The extract of the plants from Camellia sinensis (leaves), Delonix regia (flowers), Holarrhena antidysenterica (bark), Lawsonia inermis (leaves), Punica granatum (rind), Terminalia chebula (fruits) and Terminalia belerica (fruits) showed a broad-spectrum of antibacterial activity with an inhibition zone size of 11 mm to 27 mm, against all the test bacteria. The extracts from the leaves of Ocimum sanctum showed better activity against the three MRSA strains. On the other hand, extracts from Allium sativum (bulb) and Citrus sinensis (rind) exhibited little or no activity, against MRSA strains. The antibacterial potency of crude extracts was determined in terms of minimum inhibitory concentration (MIC) by the tube dilution method. MIC values, of the plant extracts, ranged from 1.3 to 8.2 mg/ml, against the test bacteria. Further, the extracts from Punica granatum and Delonix regia were fractionated in benzene, acetone and methanol. Antibacterial activity was observed in acetone as well as in the methanol fractions. In vitro synergistic interaction of crude extracts from Camellia sinensis, Lawsonia inermis, Punica granatum, Terminalia chebula and Terminalia belerica was detected with tetracycline. Moreover, the extract from Camellia sinensis also showed synergism with ampicillin.TLC of the above extracts revealed the presence of major phytocompounds, like

  6. Relationship between Legionella pneumophila and Acanthamoeba polyphaga: Physiological status and susceptibility to chemical inactivation

    SciTech Connect

    Barker, J.; Farrell, I. ); Brown, M.R.W.; Collier, P.J.; Gilbert, P. )

    1992-08-01

    Survival studies were conducted on Legionella pneumophila cells that had been grown intracellulary in Acanthamoeba polyphaga and then exposed to polyhexamethylene biguanide (PHMB), benzisothiazolone (BIT), and 5-chloro-N-methylisothiazolone (CMIT). Susceptibilities were also determined for L. pneumophila grown under iron-sufficient and iron-depleted conditions. BIT was relatively ineffective against cells to PHMB and CMIT. The activities of all three biocides were greatly reduced against L. pneumophila grown in amoebae. PHMB (1 [times] MIC) gave 99.99% reductions in viability for cultures grown in broth within 6 h and no detectable survivors at 24 h but only 90 and 99.9% killing at 6 h and 24 h, respectively, for cells grown in amoebae. The antimicrobial properties of the three biocides against A. polyphaga were also determined. The majority of amoebae recovered from BIT treatment, but few, if any, survived CMIT treatment or exposure of PHMB. This study not only shows the profound effect that intra-amoebal growth has on the physiological status and antimicrobial susceptibility of L. pneumophila but also reveals PHMB to be a potential biocide for effective water treatment. In this respect, PHMB has significant activity, below its recommended use concentrations, against both the host amoeba and L. pneumophila.

  7. Ultrastructural demonstration of spirochetal antigens in synovial fluid and synovial membrane in chronic Lyme disease: possible factors contributing to persistence of organisms.

    PubMed

    Nanagara, R; Duray, P H; Schumacher, H R

    1996-10-01

    To perform the first systematic electronmicroscopic (EM) and immunoelectron microscopy (IEM) study of the pathological changes and the evidence of spirochete presence in synovial membranes and synovial fluid (SF) cells of patients with chronic Lyme arthritis. EM examination was performed on four synovial membrane and eight SF cell samples from eight patients with chronic Lyme disease. Spirochetal antigens in the samples were sought by IEM using monoclonal antibody to Borrelia burgdorferi outer surface protein A (OspA) as the immunoprobe. Prominent ultrastructural findings were surface fibrin-like material, thickened synovial lining cell layer and signs of vascular injury. Borrelia-like structures were identified in all four synovial membranes and in two of eight SF cell samples. The presence of spirochetal antigens was confirmed by IEM in all four samples studied (one synovial membrane and three SF cell samples). OspA labelling was in perivascular areas, deep synovial stroma among collagen bundles, and in vacuoles of fibroblasts in synovial membranes; and in cytophagosomes of mononuclear cells in SF cell samples. Electron microscopy adds further evidence for persistence of spirochetal antigens in the joint in chronic Lyme disease. Locations of spirochetes or spirochetal antigens both intracellulary and extracellulary in deep synovial connective tissue as reported here suggest sites at which spirochaetes may elude host immune response and antibiotic treatment.

  8. Endosymbiotic heterocystous cyanobacteria synthesize different heterocyst glycolipids than free-living heterocystous cyanobacteria.

    PubMed

    Schouten, Stefan; Villareal, Tracy A; Hopmans, Ellen C; Mets, Anchelique; Swanson, Kathleen M; Sinninghe Damsté, Jaap S

    2013-01-01

    The heterocysts of limnetic nitrogen-fixing filamentous cyanobacteria contain unique glycolipids in their cell wall that create the distinctive gas impermeability of the heterocyst cell wall as well as serve as biomarker lipids for these microbes. It has been assumed that marine free-living and endosymbiotic cyanobacteria synthesize the same glycolipids although they have not been investigated in any detail. Here we report the glycolipid composition of several marine free-living heterocystous cyanobacteria as well as the heterocystous endosymbiont Richelia intracellularis found in the biogeochemically important diatoms Hemiaulus hauckii and Hemiaulus membranaceus. In the marine cyanobacteria Nostoc muscorum and Calothrix sp., we detected the same glycolipids as found in freshwater representatives of these genera. However, we did not detect these glycolipids in the Hemiaulus-Richelia association. Instead, we identified glycolipids which comprised a C₅ sugar, ribose, rather than the C₆ sugars normally encountered in glycolipids of free-living cyanobacteria. In addition, the glycolipids had slightly longer chain lengths (C₃₀ and C₃₂ versus C₂₆ and C₂₈) in the aglycone moiety. The different glycolipid composition of the marine endosymbotic heterocystous cyanobacteria compared to their free-living counterparts may be an adaptation to the high intracellular O₂ concentrations within their host. These glycolipids may provide unique tracers for the presence of these microbes in marine environments and permit exploration of the evolutionary origins of these symbioses.

  9. Actinide Biocolloid Formation in Brine by Halophilic Bacteria

    SciTech Connect

    Gillow, J.B.; Francis, A.J.; Dodge, C.J.; Harris, R.; Beveridge, T.J.; Brady, P.V.; Papenguth, H.W.

    1999-07-28

    We examined the ability of a halophilic bacterium (WFP 1A) isolated from the Waste Isolation Pilot Plant (WIPP) site to accumulate uranium in order to determine the potential for biocolloid facilitated actinide transport. The bacterial cell Surface functional groups involved in the complexation of the actinide were determined by titration. Uranium, added as uranyl nitrate, was removed from solution at pH 5 by cells but at pH 7 and 9 very little uranium was removed due to its limited volubility. Although present as soluble species, uranyl citrate at pH 5, 7, and 9, and uranyl carbonate at pH 9 were not removed by the bacterium because they were not bioavailable due to their neutral or negative charge. Addition of uranyl EDTA to brine at pH 5, 7, and 9 resulted in the immediate precipitation of U. Transmission electron microscopy (TEM) and energy dispersive X-ray spectroscopy (EDS) analysis revealed that uranium was not only associated with the cell surface but also accumulated intracellulary as uranium-enriched granules. Extended X-ray absorption fine structure (EXAFS) analysis, of the bacterial cells indicated the bulk sample contained more than one uranium phase. Nevertheless these results show the potential for the formation of actinide bearing bacterial biocolloids that are strictly regulated by the speciation and bioavailability of the actinide.

  10. Novel Lipid Biomarkers for Past Oceanic N2 Fixation

    NASA Astrophysics Data System (ADS)

    Bale, N. J.; Hopmans, E. C.; Villareal, T. A.; Zell, C. I.; Sinninghe Damsté, , J.; Schouten, S.

    2014-12-01

    Nitrogen-fixing cyanobacteria play important roles in the biogeochemical cycles of aquatic systems. Both heterocystous and non-heterocystous N2-fixing cyanobacteria are symbiotic with marine diatoms and thrive in low nutrient environments. These associations are significant exporters of carbon to the deep-sea, but suitable tracers for reconstructing their importance in past environments are lacking. We recently analyzed the heterocyst glycolipids (HGs) of the heterocystous Richelia intracellularis symbiont of the marine diatoms Hemiaulus hauckii and H. membranaceus and found unique C5 glycolipids with C30-32 carbon chains, a structure different from the C6 glycolipids detected in freshwater heterocystous cyanobacteria. We developed a high performance liquid chromatography/ multiple reaction monitoring mass spectrometry (HPLC/MS) method specific for trace analysis of long chain C5 HGs and applied it to suspended particulate matter (SPM) and surface sediment from the Amazon plume, a region known to harbor marine diatoms carrying heterocystous cyanobacteria as endosymbionts. C5 HGs were detected in both SPM and sediments demonstrating their biomarker potential. They were not detected in SPM or sediment from freshwater settings in the region. Rather, limnetic SPM and sediments contained C6 HGs which are established biomarkers for free-living heterocystous cyanobacteria. Glycolipids have been found preserved in sediments of up to 49 Ma old. Our development of the C5 biomarkers has the potential to improve our knowledge of the contribution of symbiotic cyanobacteria to the paleo-N-cycle.

  11. Effect of entF deletion on iron acquisition and erythritol metabolism by Brucella abortus 2308.

    PubMed

    Jain, Neeta; Rodriguez, Araceli C; Kimsawatde, Gade; Seleem, Mohamed N; Boyle, Stephen M; Sriranganathan, Nammalwar

    2011-03-01

    Brucella abortus has been shown to produce two siderophores: 2,3-dihydroxybenzoic acid (2,3-DHBA) and brucebactin. Previous studies on Brucella have shown that 2,3-DHBA is associated with erythritol utilization and virulence in pregnant ruminants. The biosynthetic pathway and role of brucebactin are not known and the only gene shown to be involved so far is entF. Using cre-lox methodology, an entF mutant was created in wild-type B. abortus 2308. Compared with the wild-type strain, the ΔentF strain showed significant growth inhibition in iron minimal media that became exacerbated in the presence of an iron chelator. For the first time, we have demonstrated the death of the ΔentF strain under iron-limiting conditions in the presence of erythritol. Addition of FeCl(3) restored the growth of the ΔentF strain, suggesting a significant role in iron acquisition. Further, complementation of the ΔentF strain using a plasmid containing an entF gene suggested the absence of any polar effects. In contrast, there was no significant difference in survival and growth between the ΔentF and wild-type strains grown in the murine macrophage cell line J774A.1, suggesting that an alternate iron acquisition pathway is present in Brucella when grown intracellulary.

  12. [Prion neuroinfections].

    PubMed

    Tichý, J

    2004-01-01

    Prion neuroinfections represent an emergent problem namely after the epidemic brake out of the bovine spongiform encephalopathies, which culminated in 1993. Isolated cases can still emerge. The next case was the discovery of a new variant of Creutzfeldt-Jakob disease in the comparatively young persons with proved alimentary infection route--about 140 cases has been described. Physiological prions are glycoproteins, affixed to the rafts of cell membranes (namely in neurons) by glycosylphosphatidylinositol anchor. They act as signalling molecules and participate in the cooper metabolism. Despite the intensive study in many prestigious laboratories, the way how the pathogenic isoform of prions, given by the prevalence of beta-conformations in the polypeptide chain develops has not been proved yet. Pathogenic isoforms are formed intracellulary after the precedent endocytosis. Aggregations of abnormal prion molecules build up prion amyloid deposits in the form of fibrils or plaques. The loss of neurons brings about the neurological symptoms, which are not directly related to the amounts of amyloid deposits. Characteristic findings of spongiosis and the immunohistochemical evidence of protease-resistant prions belong to the only reliable tests. Reliable intravital test in the blood or urine is not yet available. All forms of human and animal transmissible spongiform encephalopathies develop only in genetically sensitive individuals. A brief overview of clinical forms and some biochemical and genetic aspects of the disease are given.

  13. Diversity and distribution of diazotrophic communities in the South China Sea deep basin with mesoscale cyclonic eddy perturbations.

    PubMed

    Zhang, Yao; Zhao, Zihao; Sun, Jun; Jiao, Nianzhi

    2011-12-01

    The South China Sea (SCS) is an oligotrophic subtropical marginal ocean with a deep basin and a permanently stratified central gyre. Upwelling and nitrogen fixation provide new nitrogen for primary production in the SCS. This study was aimed at an investigation of phylogenetic diversity and quantification of the diazotroph community in the SCS deep basin, which is characterized by frequent mesoscale eddies. The diazotroph community had a relatively low diversity but a distinct spatial heterogeneity of diversity in the SCS deep basin. The potential for nitrogen fixation consistently occurred during cyclonic eddies, although upwelling of nutrient-replete deep water might have alleviated nitrogen limitation in the SCS. However, diazotrophic proteobacteria were dominant, but neither Trichodesmium nor heterocystous cyanobacterial diatom symbionts. Quantitative PCR analysis using probe-primer sets developed in this study revealed that the nif H gene of the two dominant alpha- and gammaproteobacterial groups was at the highest abundance (up to 10(4) to 10(5)  copies L(-1) ). Trichodesmium thiebautii was detected with an average density of 10(2)  trichomes L(-1) in the euphotic waters, while Richelia intracellularis was observed sporadically under the microscope. The unicellular cyanobacterial groups A and B were not detected in our libraries. Our results suggested that diazotrophic proteobacteria were significant components potentially contributing to nitrogen fixation in this oligotrophic marginal ocean ecosystem.

  14. Assessment of antioxidant capacity and cytotoxicity of selected Malaysian plants.

    PubMed

    Ling, Lai Teng; Radhakrishnan, Ammu Kutty; Subramaniam, Thavamanithevi; Cheng, Hwee Ming; Palanisamy, Uma D

    2010-04-01

    Thirteen Malaysian plants; Artocarpus champeden, Azadirachta indica, Fragaria x ananassa, Garcinia mangostana, Lawsonia inermis, Mangifera indica, Nephelium lappaceum, Nephelium mutobile, Peltophorum pterocarpum, Psidium guajava and Syzygium aqueum, selected for their use in traditional medicine, were subjected to a variety of assays. Antioxidant capability, total phenolic content, elemental composition, as well as it cytotoxity to several cell lines of the aqueous and ethanolic extracts from different parts of these selected Malaysian plants were determined. In general, the ethanolic extracts were better free radical scavengers than the aqueous extracts and some of the tested extracts were even more potent than a commercial grape seed preparation. Similar results were seen in the lipid peroxidation inhibition studies. Our findings also showed a strong correlation of antioxidant activity with the total phenolic content. These extracts when tested for its heavy metals content, were found to be below permissible value for nutraceutical application. In addition, most of the extracts were found not cytotoxic to 3T3 and 4T1 cells at concentrations as high as 100 microg/mL. We conclude that although traditionally these plants are used in the aqueous form, its commercial preparation could be achieved using ethanol since a high total phenolic content and antioxidant activity is associated with this method of preparation.

  15. Gut Health of Pigs: Challenge Models and Response Criteria with a Critical Analysis of the Effectiveness of Selected Feed Additives — A Review

    PubMed Central

    Adewole, D. I.; Kim, I. H.; Nyachoti, C. M.

    2016-01-01

    The gut is the largest organ that helps with the immune function. Gut health, especially in young pigs has a significant benefit to health and performance. In an attempt to maintain and enhance intestinal health in pigs and improve productivity in the absence of in-feed antibiotics, researchers have evaluated a wide range of feed additives. Some of these additives such as zinc oxide, copper sulphate, egg yolk antibodies, mannan-oligosaccharides and spray dried porcine plasma and their effectiveness are discussed in this review. One approach to evaluate the effectiveness of these additives in vivo is to use an appropriate disease challenge model. Over the years, researchers have used a number of challenge models which include the use of specific strains of enterotoxigenic Escherichia coli, bacteria lipopolysaccharide challenge, oral challenge with Salmonella enteric serotype Typhimurium, sanitation challenge, and Lawsonia intercellularis challenge. These challenge models together with the criteria used to evaluate the responses of the animals to them are also discussed in this review. PMID:26954144

  16. Vibriocidal activity of certain medicinal plants used in Indian folklore medicine by tribals of Mahakoshal region of central India

    PubMed Central

    Sharma, Anjana; Patel, Virendra Kumar; Chaturvedi, Animesh Navin

    2009-01-01

    Objectives: Screening of the medicinal plants and determination of minimum inhibitory concentration (MIC) against Vibrio cholerae and Vibrio parahaemolyticus. Materials and Methods: A simple in vitro screening assay was employed for the standard strain of Vibrio cholerae, 12 isolates of Vibrio cholerae non-O1, and Vibrio parahaemolyticus. Aqueous and organic solvent extracts of different parts of the plants were investigated by using the disk diffusion method. Extracts from 16 medicinal plants were selected on account of the reported traditional uses for the treatment of cholera and gastrointestinal diseases, and they were assayed for vibriocidal activities. Results: The different extracts differed significantly in their vibriocidal properties with respect to different solvents. The MIC values of the plant extracts against test bacteria were found to be in the range of 2.5-20 mg/ml. Conclusions: The results indicated that Lawsonia inermis, Saraca indica, Syzygium cumini, Terminalia belerica, Allium sativum, and Datura stramonium served as broad-spectrum vibriocidal agents. PMID:20442821

  17. The effect of methanolic extract of Tamarindus indica Linn. on the growth of clinical isolates of Burkholderia pseudomallei.

    PubMed

    Muthu, Shankar Esaki; Nandakumar, Subhadra; Rao, Usha Anand

    2005-12-01

    Burkholderia pseudomallei (Pseudomonas pseudomallei) causes melioidosis, a life-threatening infection common among paddy cultivators in Southeast Asian countries. No plant materials have been investigated for its activity against B. pseudomallei. Therefore, a preliminary study was carried out using disc diffusion and minimum inhibitory concentration (MIC) methods to evaluate the anti-B. pseudomallei activity of five Indian medicinal plants documented to have been used for several ailments in the ancient Indian scriptures. The leaf extracts of Tamarindus indica, Lawsonia inermis, and Hibiscus rosa-sinensis, the rhizome extracts of Curcuma longa and the seeds of Vigna radiata were prepared using methanol as solvent. The disc diffusion and MIC methods were used to assess the anti-B. pseudomallei activity of the plants tested. Only methanol leaf extracts of Tamarindus indica exhibited anti-B. pseudomallei activity starting from disc concentrations of 150 mug by the disc diffusion method. The other plants failed to show any zone of inhibition. MIC assay revealed that the MIC and minimum bactericidal concentration (MBC) for B. pseudomallei were 125 mug/ml. Our preliminary finding showed that methanolic extracts of Tamarindus indica has anti-B. pseudomallei inhibitory potentials under in vitro conditions. Extensive animal studies may be required before investigating the role of Tamarindus indica for treating melioidosis. PMID:16518004

  18. The effect of methanolic extract of Tamarindus indica Linn. on the growth of clinical isolates of Burkholderia pseudomallei.

    PubMed

    Muthu, Shankar Esaki; Nandakumar, Subhadra; Rao, Usha Anand

    2005-12-01

    Burkholderia pseudomallei (Pseudomonas pseudomallei) causes melioidosis, a life-threatening infection common among paddy cultivators in Southeast Asian countries. No plant materials have been investigated for its activity against B. pseudomallei. Therefore, a preliminary study was carried out using disc diffusion and minimum inhibitory concentration (MIC) methods to evaluate the anti-B. pseudomallei activity of five Indian medicinal plants documented to have been used for several ailments in the ancient Indian scriptures. The leaf extracts of Tamarindus indica, Lawsonia inermis, and Hibiscus rosa-sinensis, the rhizome extracts of Curcuma longa and the seeds of Vigna radiata were prepared using methanol as solvent. The disc diffusion and MIC methods were used to assess the anti-B. pseudomallei activity of the plants tested. Only methanol leaf extracts of Tamarindus indica exhibited anti-B. pseudomallei activity starting from disc concentrations of 150 mug by the disc diffusion method. The other plants failed to show any zone of inhibition. MIC assay revealed that the MIC and minimum bactericidal concentration (MBC) for B. pseudomallei were 125 mug/ml. Our preliminary finding showed that methanolic extracts of Tamarindus indica has anti-B. pseudomallei inhibitory potentials under in vitro conditions. Extensive animal studies may be required before investigating the role of Tamarindus indica for treating melioidosis.

  19. Retroviral reverse transcriptase inhibitory activity in Thai herbs and spices: screening with Moloney murine leukemia viral enzyme.

    PubMed

    Suthienkul, O; Miyazaki, O; Chulasiri, M; Kositanont, U; Oishi, K

    1993-12-01

    Fifty-seven Thai herbs and spices were examined for their retroviral reverse transcriptase inhibitory activity. All herbs and spices were extracted with hot-water and methanol. Reverse transcriptase inhibitory activity of the extracts was determined by using Moloney Murine Leukemia Virus reverse transcriptase (M-MuLV-RT) reacted with 3H-dTTP and radioactivity measured with a scintillation counter. Eighty-one per cent (46/57) of hot-water extracts and 54% (31/57) of methanol extracts showed inhibitory activities. At a concentration of 125 micrograms/ml, 13% (6/46) of hot-water extracts, namely Eugenia caryophyllus Bullock et Harrison, Phyllanthus urinaria Linn., Terminalia belerica Roxb., Nelumbo nucifera Gaertn., Psidium guajava Linn. and Lawsonia inermis Linn., had a relative inhibitory ratio (IR) over 50%. They showed ratios of 100%, 91%, 75%, 74%, 61% and 60%, respectively. For methanol extracts, only 10% (3/31) had IR values over 50%. They were T. belerica, E. caryophyllus and N. nucifera which exhibited IR values of 83%, 54% and 54%, respectively. PMID:7524165

  20. Synthesis, spectral characterization, molecular structure and pharmacological studies of N'-(1, 4-naphtho-quinone-2yl) isonicotinohyWdrazide

    NASA Astrophysics Data System (ADS)

    Kavitha Rani, P. R.; Fernandez, Annette; George, Annie; Remadevi, V. K.; Sudarsanakumar, M. R.; Laila, Shiny P.; Arif, Muhammed

    2015-01-01

    A simple and efficient procedure was employed for the synthesis of N'-(1,4-naphtho-quinone-2-yl) isonicotinohydrazide (NIH) by the reaction of 2-hydroxy-1,4-naphthaquinone (lawsone) and isonicotinoyl hydrazine in methanol using ultrasonic irradiation. Lawsone is the principal dye, isolated from the leaves of henna (Lawsonia inermis). Structural modification was done on the molecule aiming to get a more active derivative. The structure of the parent compound and the derivative was characterized by elemental analyses, infrared, electronic, 1H, 13C NMR and GC-MS spectra. The fluorescence spectral investigation of the compound was studied in DMSO and ethanol. Single crystal X-ray diffraction studies reveal that NIH crystallizes in monoclinic space group. The DNA cleavage study was monitored by gel electrophoresis method. The synthesized compound was found to have significant antioxidant activity against DPPH radical (IC50 = 58 μM). The in vitro cytotoxic studies of the derivative against two human cancer cell lines MCF-7 (human breast cancer) and HCT-15 (human colon carcinoma cells) using MTT assay revealed that the compound exhibited higher cytotoxic activity with a lower IC50 value indicating its efficiency in killing the cancer cells even at low concentrations. These results suggest that the structural modifications performed on lawsone could be considered a good strategy to obtain a more active drug.

  1. Determination of heavy metal contents by atomic absorption spectroscopy (AAS) in some medicinal plants from Pakistani and Malaysian origin.

    PubMed

    Akram, Sobia; Najam, Rahila; Rizwani, Ghazala H; Abbas, Syed Atif

    2015-09-01

    This study depicts a profile of existence of heavy metals (Cu, Ni, Zn, Cd, Hg, Mn, Fe, Na, Ca, and Mg) in some important herbal plants like (H. Integrifolia, D. regia, R. communis, C. equisetifolia, N. oleander, T. populnea, M. elengi, H. schizopetalus, P. pterocarpum) from Pakistan and an antidiabetic Malaysian herbal drug product containing (Punica granatum L. (Mast) Hook, Momordica charantia L., Tamarindus indica L., Lawsonia inermis L.) using atomic absorption spectrophotometer. Heavy metals in these herbal plants and Malaysian product were in the range of 0.02-0.10 ppm of Cu, 0.00-0.02 ppm of Ni, 0.02-0.29 ppm of Zn, 0.00-0.04 ppm of Cd, 0.00-1.33 ppm of Hg, 0.00-0.54 ppm of Mn, 0.22-3.16 ppm of Fe, 0.00-9.17 ppm of Na, 3.27-15.63 ppm of Ca and 1.85-2.03 ppm of Mg. All the metals under study were within the prescribed limits except mercury. Out of 10 medicinal plants/product under study 07 were beyond the limit of mercury permissible limits. Purpose of this study is to determine heavy metals contents in selected herbal plants and Malaysian product, also to highlight the health concerns related to the presence of toxic levels of heavy metals.

  2. Mitochondrial malate dehydrogenase and malic enzyme of a filarial worm Setaria digitata: some properties and effects of drugs and herbal extracts.

    PubMed

    Banu, M J; Nellaiappan, K; Dhandayuthapani, S

    1992-06-01

    Mitochondrial malate dehydrogenase (mMDH) and malic enzyme (mME) of a filarial worm Setaria digitata were studied. mMDH exhibited the highest activities in the oxidation and reduction reactions at pH 9.5 and pH 6.2, respectively, while mME did so in the malate decarboxylation reaction at pH 6.8. mME showed no detectable activity on the pyruvate carboxylation direction. The Km values for malate (1.7 mM) and oxaloacetate (0.17 mM) and the ratio of Vmax oxidation: Vmax reduction (2.73) tend to favor the oxaloacetate reduction by mMDH. mME showed a relatively high Km value of 8.3 mM, for malate decarboxylation. A drug, diethylcarbamazine citrate (DEC-C), did not change appreciably the activity of either mMDH or mME, while filarin (a drug of herbal origin) effectively inhibited mMDH. The leaf extracts of Ocimum sanctum, Lawsonia inermis and Calotropis gigantea and leaf and flower extracts of Azadirachta indica were, however, found to inhibit both mMDH and mME.

  3. Hepatoprotective Potential of Some Local Medicinal Plants against 2-Acetylaminoflourene-Induced Damage in Rat

    PubMed Central

    Adetutu, Adewale; Olorunnisola, Olubukola S.

    2013-01-01

    The in vivo micronucleus assay was used to examine the anticlastogenic effects of crude extracts of Bridelia ferruginea, Vernonia amygdalina, Tridax procumbens, Ocimum gratissimum, and Lawsonia inermis in Wistar albino rats. Extracts of doses of 100 mg/kg body weight were given to rats in five groups for seven consecutive days followed by a single dose of 2-AAF (0.5 mmol/kg body weight). The rats were sacrificed after 24 hours and their bone marrow smears were prepared on glass slides stained with Giemsa. The micronucleated polychromatic erythrocyte cells (mPCEs) were thereafter recorded. The hepatoprotective effects of the plant extracts against 2-AAF-induced liver toxicity in rats were evaluated by monitoring the levels of alkaline phosphatase (ALP), gamma glutamyl transferase (GGT), and histopathological analysis. The results of the 2-AAF-induced liver toxicity experiments showed that rats treated with the plant extracts (100 mg/kg) showed a significant decrease in mPCEs as compared with the positive control. The rats treated with the plant extracts did not show any significant change in the concentration of ALP and GGT in comparison with the negative control group whereas the 2-AAF group showed a significant increase (P < 0.05) in these parameters. Some of the leaf extracts also showed protective effects against histopathological alterations. This study suggests that the leaf extracts have hepatoprotective potential, thereby justifying their ethnopharmacological uses. PMID:24163694

  4. A preliminary pilot survey on head lice, pediculosis in Sharkia Governorate and treatment of lice with natural plant extracts.

    PubMed

    El-Basheir, Zeinab M; Fouad, Mahmoud A H

    2002-12-01

    Twelve different representative areas in Sharkia Governorate were surveyed for head lice, Pediculus humanus capitis. The pre-valence was investigated among 120 houses containing 2,448 individual, with different age, sex and socioeconomic status. Examination was done by naked eye aided with hand-lens. A total of 137 individuals were infested. Infestation rates were higher in the rural areas with low socioeconomic levels, concrete houses with over-crowded family members. Children had significantly higher infestation rates than adults. Males had lower infestation rates than females. However, the hair length and permanent hair washing were the factors accounted for both age and sex difference in prevalence of pediculosis. Head lice infestations were found all over the year, but increased in summer and spring. One hundred infested patients (90 females and 10 males) with different aged and hair length were treated with tour mixed cream from plants Lawsonia alba L. (Henna). Trigonella faemum-gracanum (Fenugreek), Hibiscus cannabinus (Hibiscus) and Artemisia cina (Wormseed). The head lice completely disappeared within a week among those patients treated by henna mixed with aqueous extract of sheah (100%) or mixed with helba (75%) or with karkada (50%).

  5. Determination of the Moisture-Sorption Isotherms and Isosteric Heat of Henna Leaves

    NASA Astrophysics Data System (ADS)

    Bennaceur, S.; Draoui, B.; Touati, B.; Benseddik, A.; Saad, A.; Bennamoun, L.

    2015-01-01

    Equilibrium moisture desorption and adsorption isotherms of Lawsonia inermis L. (commonly known as henna) leaves at temperatures of 30, 40 and 50°C with a water activity ranging from 0.057 to 0.898 were obtained by the gravimetric-static method. It was established that when the temperature of these leaves increases, their moisture content increases too with a hysteresis effect. The experimental data on the sorption of the indicated leaves were compared with the corresponding calculation data obtained with the use of the GAB, modified BET, Henderson-Thompson, modified Halsey, modified Oswin, and Peleg models. Evaluation of these models on the basis of statistical processing of the data obtained with them, including the calculus of the standard error and the correlation coefficient, has shown that the GAB and Peleg models represent sorption curves more adequately. The net isosteric heats of desorption and adsorption of henna leaves were determined by the sorption isotherms constructed using the Clausius-Clapeyron equation. An expression for predicting these thermodynamic properties of plants is proposed.

  6. Side-effects of henna and semi-permanent 'black henna' tattoos: a full review.

    PubMed

    de Groot, Anton C

    2013-07-01

    Henna, the dried and powdered leaf of Lawsonia inermis, is widely used as a dye for the skin, hair, and nails, and as an expression of body art, especially in Islamic and Hindu cultures. As it stains the skin reddish-brown, it is also called red henna. Black henna is the combination of red henna with p-phenylenediamine (PPD), and is used for temporary 'black henna tattoos'. This article provides a full review of the side-effects of topical application of red and black henna, both cutaneous (allergic and non-allergic) and systemic. Red henna appears to be generally safe, with rare instances of contact allergy and type I hypersensitivity reactions. In children with glucose-6-phosphate dehydrogenase deficiency, topical application of henna may cause life-threatening haemolysis. Black henna tattoos will induce contact allergy to its ingredient PPD at an estimated frequency of 2.5%. Once sensitized, the patients may experience allergic contact dermatitis from the use of hair dyes containing PPD. There are often cross-reactions to other hair dyes, dyes used in textiles, local anaesthetics, and rubber chemicals. The sensitization of children to PPD may have important consequences for health and later career prospects. Systemic toxicity of black henna has been reported in certain African countries.

  7. Management of corm-rot disease of Gladiolus by plant extracts.

    PubMed

    Riaz, Tariq; Nawaz Khan, Salik; Javaid, Arshad

    2010-07-01

    A pot experiment was conducted to evaluate the efficacy of aqueous extracts of six plant species, namely Azadirachta indica A. Juss. (neem), Alstonia scholaris (L.) R. Br., Lawsonia alba Lam., Allium cepa L., A. sativum L. and Zingiber officinale Roscoe, and a systemic fungicide carbendazim 50% (w/w) WP, to manage the corm-rot disease of Gladiolus (Gladiolus grandiflorus L.) caused by a fungal pathogen Fusarium oxysporum f.sp. gladioli (Massey) Snyd. & Hans. Fusarium inoculation showed 80% disease incidence with 54 disease lesions per corm. Recommended dose of the chemical fungicide carbendazim significantly reduced the disease incidence to 13% and number of lesions to six per corm. Plant extract treatments exhibited variable effects on the incidence and severity of the disease. In general, all the test plant extracts managed the corm-rot disease to some extent. Aqueous bulb extracts of A. sativum and A. cepa and the rhizome extract of Z. officinale showed better disease management potential than that of the recommended dose of carbendazim. Fusarium inoculation significantly declined shoot growth. In general, carbendazim, as well as aqueous extracts, enhanced shoot growth to variable extents as compared to the Fusarium control. PMID:19557652

  8. Nitrogen modulates phytoplankton growth in spring in the South China Sea

    NASA Astrophysics Data System (ADS)

    Chen, Yuh-ling Lee; Chen, Houng-Yung; Karl, David M.; Takahashi, Masayuki

    2004-03-01

    The present study investigated the roles of nitrogen (N) in modulating phytoplankton growth in the South China Sea (SCS), an oligotrophic marginal sea in the western Pacific Ocean, by examining (1) the in situ nitrate and nitrite concentrations [NO 3+NO 2]; (2) the [ NO3+ NO2]/[ soluble reactive phosphorus ( SRP)] in the euphotic layer; (3) phytoplankton growth in response to nutrient enrichments; and (4) the spatial distribution of nitrate-based new production and primary production and the factors controlling their dynamics. N 2-fixation rates of the planktonic cyanobacteria, Trichodesmium sp. and the diatom-symbiont Richelia intracellularis, were estimated based on the abundance of the N 2-fixers and were compared to the nitrogen demands estimated based on the nitrate uptake rate (nitrate-based new production). The comparison evaluates the relative importance of two sources of new nitrogen, the atmosphere (N 2) vs. the deep waters (NO 3-). The study was conducted on cruises in March 2000 and March 2001 when the northeastern monsoon was prevailing in the SCS. Both N and phosphorus (P) were scarce in the euphotic zone; surface [NO 3+NO 2] was in the range of 9- 309 nM and [SRP] ranged from 6 to 88 nM. Surface [NO 3+NO 2] was negatively ( p<0.01) correlated with stratification index of water column. The ratios of [NO 3+NO 2]/[SRP] were much smaller than the Redfield N/P Ratio of 16:1, suggesting a state of N-limitation. The water of the shallow shelf-slope was less stratified. Its surface nitrate concentration, chlorophyll a concentration, nitrate-based new production and total primary production were higher than those of the deep basin water. Simulated production experiments on the shelf and the basin revealed that nitrate-based new production ranged from 0.06 to 0.36 gC m -2 d-1 whereas primary production ranged from 0.12 to 0.63 gC m -2 d-1. Both productions are positively ( p<0.01) correlated with the surface nitrate concentration. The results of the enrichment

  9. Nitrogen fixation in an anticyclonic eddy in the oligotrophic North Pacific Ocean.

    PubMed

    Fong, Allison A; Karl, David M; Lukas, Roger; Letelier, Ricardo M; Zehr, Jonathan P; Church, Matthew J

    2008-06-01

    Mesoscale physical processes (for example eddies, frontal meanders and planetary waves) can play important roles in controlling ocean biogeochemistry. We examined spatial variations in upper ocean (0-100 m) nutrient inventories, N(2) fixing microorganism diversity and abundance, and rates of N(2) fixation in an anticyclonic eddy near Station ALOHA (22 degrees 45' N, 158 degrees 00' W) in the North Pacific Subtropical Gyre (NPSG). In July 2005, satellite-based sea surface altimetry and ocean color observation revealed an anticyclonic eddy with enhanced chlorophyll in the upper ocean in the vicinity of Station ALOHA. Within the eddy, near-surface ocean chlorophyll concentrations were approximately 5-fold greater than in the surrounding waters. Inventories of nitrate and phosphate in the eddy were similar to the concentrations historically observed at Station ALOHA, while silicic acid inventories were significantly depleted (one-way analysis of variance, P<0.01). Quantitative PCR determinations of nifH gene copies revealed relatively high abundances of several N(2) fixing cyanobacteria, including Trichodesmium spp., Crocosphaera watsonii and Richelia intracellularis. Reverse transcriptase PCR (RT-PCR) amplified nitrogenase (nifH) gene transcripts were cloned and sequenced to examine the diversity of active N(2) fixing microorganisms; these clone libraries were dominated by sequence-types 97%-99% identical to the filamentous cyanobacteria Trichodesmium spp. Near-surface ocean rates of N(2) fixation were 2-18 times greater (averaging 8.6+/-5.6 nmol N per l per day) than previously reported measurements at Station ALOHA. These results suggest that mesoscale physical variability can play an important role in modifying the abundances of N(2) fixing microorganisms and associated rates of N(2) fixation in open ocean ecosystems.

  10. Occurrence of Diatom-Cyanobacterial symbiotic relationship in the coastal surface waters of Northern Red Sea

    NASA Astrophysics Data System (ADS)

    khomayis, hisham; Sommer, Ulrich; Al-Aidaroos, Ali; Kürten, Benjamin; Devassy, Reny

    2013-04-01

    The phenomenon of Diatom-Cyanobacterial symbiotic association is widely noticed from different marine ecosystems around the world especially in the tropical and subtropical regions. The fixation of atmospheric nitrogen in the oligotrophic ocean is an important source of new nitrogen to surface waters, stimulating phytoplankton productivity and fueling the biological pump. The most common cyanobacterium that carry out the so called nitrogen fixation in marine system is Trichodesmium. The other major representative of this diazotrophic group is the heterocystous endosymbiotic cyanobacterium known as Richelia intracellularis which always seen in symbiotic relationship with other phytoplankton and is well-known for its capability to fix the atmospheric nitrogen in oligotrophic waters. Planktonic diatom- cyanobaterial symbioses play a significant role in the ecology and biogeochemistry of the surface oceans. Studies suggest that these particular diazotroph communities contribute annually 4.79 × 1012 gm of nitrogen to the water which comes almost one quarter of the total nitrogen input to the sea. The endosymbiont Richelia normally occupies the periplasmic space between the plasmalemma and the cell wall of the host and it provides a source of nutrient availability to the host where it is associated. They are expected to release the fixed nitrogen as either ammonium or dissolved organic nitrogen, which is then available to the particular host which lives in nutrient-starved condition. It is mainly known to be associated with the centric diatoms like Rhizosolenia, Hemialus, and Guinardia. The present observation of the Rhizosolenia-Richelia and Hemialus-Richelia symbiotic relation from the coastal waters of northern red sea implies much attention to the ecology of the particular area. While considering the oligotrophic nature of the region, the particular association helps the phytoplankton to avoid the limited growth due to the scarcity of nitrogen to a great extent. The

  11. Phytoplankton community structure in the VAHINE mesocosm experiment

    NASA Astrophysics Data System (ADS)

    Leblanc, Karine; Cornet, Véronique; Caffin, Mathieu; Rodier, Martine; Desnues, Anne; Berthelot, Hugo; Turk-Kubo, Kendra; Heliou, Jules

    2016-09-01

    The VAHINE mesocosm experiment was designed to trigger a diazotroph bloom and to follow the subsequent transfer of diazotroph-derived nitrogen (DDN) in the rest of the food web. Three mesocosms (50 m3) located inside the Nouméa lagoon (New Caledonia, southwestern Pacific) were enriched with dissolved inorganic phosphorus (DIP) in order to promote N2 fixation in these low-nutrient, low-chlorophyll (LNLC) waters. Initially, the diazotrophic community was dominated by diatom diazotroph associations (DDAs), mainly by Rhizosolenia/Richelia intracellularis, and by Trichodesmium, which fueled enough DDN to sustain the growth of other diverse diatom species and Synechococcus populations that were well adapted to limiting DIP levels. After DIP fertilization (1 µM) on day 4, an initial lag time of 10 days was necessary for the mesocosm ecosystems to start building up biomass. However, changes in community structure were already observed during this first period, with a significant drop of both Synechococcus and diatom populations, while Prochlorococcus benefited from DIP addition. At the end of this first period, corresponding to when most added DIP was consumed, the diazotroph community changed drastically and became dominated by Cyanothece-like (UCYN-C) populations, which were accompanied by a monospecific bloom of the diatom Cylindrotheca closterium. During the second period, biomass increased sharply together with primary production and N2-fixation fluxes near tripled. Diatom populations, as well as Synechococcus and nanophytoeukaryotes, showed a re-increase towards the end of the experiment, showing efficient transfer of DDN to non-diazotrophic phytoplankton.

  12. Entrance and survival of Brucella pinnipedialis hooded seal strain in human macrophages and epithelial cells.

    PubMed

    Larsen, Anett K; Nymo, Ingebjørg H; Briquemont, Benjamin; Sørensen, Karen K; Godfroid, Jacques

    2013-01-01

    Marine mammal Brucella spp. have been isolated from pinnipeds (B. pinnipedialis) and cetaceans (B. ceti) from around the world. Although the zoonotic potential of marine mammal brucellae is largely unknown, reports of human disease exist. There are few studies of the mechanisms of bacterial intracellular invasion and multiplication involving the marine mammal Brucella spp. We examined the infective capacity of two genetically different B. pinnipedialis strains (reference strain; NTCT 12890 and a hooded seal isolate; B17) by measuring the ability of the bacteria to enter and replicate in cultured phagocytes and epithelial cells. Human macrophage-like cells (THP-1), two murine macrophage cell lines (RAW264.7 and J774A.1), and a human malignant epithelial cell line (HeLa S3) were challenged with bacteria in a gentamicin protection assay. Our results show that B. pinnipedialis is internalized, but is then gradually eliminated during the next 72-96 hours. Confocal microscopy revealed that intracellular B. pinnipedialis hooded seal strain colocalized with lysosomal compartments at 1.5 and 24 hours after infection. Intracellular presence of B. pinnipedialis hooded seal strain was verified by transmission electron microscopy. By using a cholesterol-scavenging lipid inhibitor, entrance of B. pinnipedialis hooded seal strain in human macrophages was significantly reduced by 65.8 % (± 17.3), suggesting involvement of lipid-rafts in intracellular entry. Murine macrophages invaded by B. pinnipedialis do not release nitric oxide (NO) and intracellular bacterial presence does not induce cell death. In summary, B. pinnipedialis hooded seal strain can enter human and murine macrophages, as well as human epithelial cells. Intracellular entry of B. pinnipedialis hooded seal strain involves, but seems not to be limited to, lipid-rafts in human macrophages. Brucella pinnipedialis does not multiply or survive for prolonged periods intracellulary. PMID:24376851

  13. Copper-free azide-alkyne cycloaddition of targeting peptides to porous silicon nanoparticles for intracellular drug uptake.

    PubMed

    Wang, Chang-Fang; Mäkilä, Ermei M; Kaasalainen, Martti H; Liu, Dongfei; Sarparanta, Mirkka P; Airaksinen, Anu J; Salonen, Jarno J; Hirvonen, Jouni T; Santos, Hélder A

    2014-01-01

    Porous silicon (PSi) has been demonstrated as a promising drug delivery vector for poorly water-soluble drugs. Here, a simple and efficient method based on copper-free click chemistry was used to introduce targeting moieties to PSi nanoparticles in order to enhance the intracellular uptake and tumor specific targeting hydrophobic drug delivery. Two RGD derivatives (RGDS and iRGD) with azide-terminated groups were conjugated to bicyclononyne-functionalized PSi nanoparticles via copper-free azide-alkyne cycloaddition. The surface functionalization was performed in aqueous solution at 37 °C for 30 min resulting in conjugation efficiencies of 15.2 and 3.4% (molar ratios) and the nanoparticle size increased from 165.6 nm to 179.6 and 188.8 nm for RGDS and iRGD, respectively. The peptides modification enhanced the cell uptake efficiency of PSi nanoparticles in EA.hy926 cells. PSi-RGDS and PSi-iRGD nanoparticles loaded with sorafenib showed a similar trend for the in vitro antiproliferation activity compared to sorafenib dissolved in dimethyl sulfoxide. Furthermore, sorafenib-loaded PSi-RGDS deliver the drug intracellulary efficiently due to the higher surface conjugation ratio, resulting in enhanced in vitro antiproliferation effect. Our results highlight the surface functionalization methodology for PSi nanoparticles applied here as a universal method to introduce functional moieties onto the surface of PSi nanoparticles and demonstrate their potential active targeting properties for anticancer drug delivery.

  14. Free heme and sickle hemoglobin polymerization

    NASA Astrophysics Data System (ADS)

    Uzunova, Veselina V.

    This work investigates further the mechanism of one of the most interesting of the protein self-assembly systems---the polymerization of sickle hemoglobin and the role of free heme in it. Polymerization of sickle hemoglobin is the primary event in the pathology of a chronic hemolytic condition called sickle cell anemia with complex pathogenesis, unexplained variability and symptomatic treatment. Auto-oxidation develops in hemoglobin solutions exposed to room temperature and causes release of ferriheme. The composition of such solutions is investigated by mass spectrometry. Heme dimers whose amount corresponds to the initial amounts of heme released from the protein are followed. Differences in the dimer peak height are established for hemoglobin variants A, S and C and depending on the exposure duration. The effects of free heme on polymerization kinetics are studied. Growth rates and two characteristic parameters of nucleation are measured for stored Hb S. After dialysis of polymerizing solutions, no spherulites are detected at moderately high supersaturation and prolonged exposure times. The addition of 0.16-0.26 mM amounts of heme to dialyzed solutions leads to restoration of polymerization. The measured kinetic parameters have higher values compared to the ones before dialysis. The amount of heme in non-dialyzed aged solution is characterized using spectrophotometry. Three methods are used: difference in absorbance of dialyzed and non-dialyzed solutions, characteristic absorbance of heme-albumin complex and absorbance of non-dialyzed solutions with added potassium cyanide. The various approaches suggest the presence of 0.12 to 0.18 mM of free ferriheme in such solutions. Open questions are whether the same amounts of free heme are present in vivo and whether the same mechanism operates intracellulary. If the answer to those questions is positive, then removal of free heme from erythrocytes can influence their readiness to sickle.

  15. Entrance and Survival of Brucella pinnipedialis Hooded Seal Strain in Human Macrophages and Epithelial Cells

    PubMed Central

    Briquemont, Benjamin; Sørensen, Karen K.; Godfroid, Jacques

    2013-01-01

    Marine mammal Brucella spp. have been isolated from pinnipeds (B. pinnipedialis) and cetaceans (B. ceti) from around the world. Although the zoonotic potential of marine mammal brucellae is largely unknown, reports of human disease exist. There are few studies of the mechanisms of bacterial intracellular invasion and multiplication involving the marine mammal Brucella spp. We examined the infective capacity of two genetically different B. pinnipedialis strains (reference strain; NTCT 12890 and a hooded seal isolate; B17) by measuring the ability of the bacteria to enter and replicate in cultured phagocytes and epithelial cells. Human macrophage-like cells (THP-1), two murine macrophage cell lines (RAW264.7 and J774A.1), and a human malignant epithelial cell line (HeLa S3) were challenged with bacteria in a gentamicin protection assay. Our results show that B. pinnipedialis is internalized, but is then gradually eliminated during the next 72 – 96 hours. Confocal microscopy revealed that intracellular B. pinnipedialis hooded seal strain colocalized with lysosomal compartments at 1.5 and 24 hours after infection. Intracellular presence of B. pinnipedialis hooded seal strain was verified by transmission electron microscopy. By using a cholesterol-scavenging lipid inhibitor, entrance of B. pinnipedialis hooded seal strain in human macrophages was significantly reduced by 65.8 % (± 17.3), suggesting involvement of lipid-rafts in intracellular entry. Murine macrophages invaded by B. pinnipedialis do not release nitric oxide (NO) and intracellular bacterial presence does not induce cell death. In summary, B. pinnipedialis hooded seal strain can enter human and murine macrophages, as well as human epithelial cells. Intracellular entry of B. pinnipedialis hooded seal strain involves, but seems not to be limited to, lipid-rafts in human macrophages. Brucella pinnipedialis does not multiply or survive for prolonged periods intracellulary. PMID:24376851

  16. Preliminary evaluation of hepatoprotective potential of the polyherbal formulation

    PubMed Central

    Arka, Ghosh; Anindita, Kundu; Ankit, Seth; Kumar, Singh Anil; Kumar, Maurya Santosh

    2015-01-01

    Aim: The aim of this study was to investigate the antioxidant and hepatoprotective effects of the polyherbal formulation (PHF)containing Cajanus cajan (L.)Millsp., Lawsonia inermis L. Linn, Mimosa pudica L., Uraria picta (Jacq.)DC. and Operculina turpethum (L.)Silva Manso on carbon tetrachloride (CCl4)induced acute liver damage in albino rats. Materials and Methods: The groups of animals were administered with PHF at the doses 100, 200 and 400 mg/kg b.w. (per oral [p.o.])once in a day for 7 days and at day 6th and 7th the animals were administrated with Carbon tetrachloride (1.0 mL/kg b.w. 50% v/v with olive oil,; p.o.). The effect of PHF on serum glutamine pyruvate transaminase (SGPT), serum glutamine oxaloacetate transaminase, alkaline phosphatase (ALP)and total bilirubin were determined in CCl4 - induced hepatotoxicity in rats. Further, the effects of PHF on glutathione (GSH), superoxide dismutase (SOD)level and lipid peroxidation (LPO)activity were also investigated. Results: The results demonstrated that PHF (400 mg/kg b.w.)significantly reduces the CCl4 induced increase in level of serum SGPT, serum ALP and total bilirubin. PHF (400 mg/kg b.w.)prevents the depletion level of GSH and decrease in the activity of SOD in CCl4 -induced liver injury in rats. In addition, PHF also showed a significant decrease in the LPO levels signifying the potent antioxidant activity. Conclusion: All our findings suggest that PHF could protect the liver cells from CCl4 - induced liver damages and the mechanism may be through the anti-oxidative effect of PHF. PMID:26401397

  17. Systematic analysis of in vitro photo-cytotoxic activity in extracts from terrestrial plants in Peninsula Malaysia for photodynamic therapy.

    PubMed

    Ong, Cheng Yi; Ling, Sui Kiong; Ali, Rasadah Mat; Chee, Chin Fei; Samah, Zainon Abu; Ho, Anthony Siong Hock; Teo, Soo Hwang; Lee, Hong Boon

    2009-09-01

    One hundred and fifty-five extracts from 93 terrestrial species of plants in Peninsula Malaysia were screened for in vitro photo-cytotoxic activity by means of a cell viability test using a human leukaemia cell-line HL60. These plants which can be classified into 43 plant families are diverse in their type of vegetation and their natural habitat in the wild, and may therefore harbour equally diverse metabolites with potential pharmaceutical properties. Of these, 29 plants, namely three from each of the Clusiaceae, Leguminosae, Rutaceae and Verbenaceae families, two from the Piperaceae family and the remaining 15 are from Acanthaceae, Apocynaceae, Bignoniaceae, Celastraceae, Chrysobalanaceae, Irvingiaceae, Lauraceae, Lythraceae, Malvaceae, Meliaceae, Moraceae, Myristicaceae, Myrsinaceae, Olacaceae and Sapindaceae. Hibiscus cannabinus (Malvaceae), Ficus deltoidea (Moraceae), Maranthes corymbosa (Chrysobalanaceae), Micromelum sp., Micromelum minutum and Citrus hystrix (Rutaceae), Cryptocarya griffithiana (Lauraceae), Litchi chinensis (Sapindaceae), Scorodocarpus bornensis (Olacaceae), Kokoona reflexa (Celastraceae), Irvingia malayana (Irvingiaceae), Knema curtisii (Myristicaceae), Dysoxylum sericeum (Meliaceae), Garcinia atroviridis, Garcinia mangostana and Calophyllum inophyllum (Clusiaceae), Ervatamia hirta (Apocynaceae), Cassia alata, Entada phaseoloides and Leucaena leucocephala (Leguminosae), Oroxylum indicum (Bignoniaceae), Peronema canescens,Vitex pubescens and Premna odorata (Verbenaceae), Piper mucronatum and Piper sp. (Piperaceae), Ardisia crenata (Myrsinaceae), Lawsonia inermis (Lythraceae), Strobilanthes sp. (Acanthaceae) were able to reduce the in vitro cell viability by more than 50% when exposed to 9.6J/cm(2) of a broad spectrum light when tested at a concentration of 20 microg/mL. Six of these active extracts were further fractionated and bio-assayed to yield four photosensitisers, all of which are based on the pheophorbide-a and -b core structures

  18. Cytotoxicity of lawsone and cytoprotective activity of antioxidants in catalase mutant Escherichia coli.

    PubMed

    Sauriasari, Rani; Wang, Da-Hong; Takemura, Yoko; Tsutsui, Ken; Masuoka, Noriyoshi; Sano, Kuniaki; Horita, Masako; Wang, Bing-Ling; Ogino, Keiki

    2007-06-01

    Lawsone is an active naphthoquinone derivative isolated from henna (Lawsonia inermis L.), a widely used hair dye. Previous study on the toxicity of lawsone remains unclear since the involvement of oxidative stress and the kind of ROS (reactive oxygen species) involved have not been fully resolved yet. This present study reports the cytotoxic effects of lawsone and henna. We carried out CAT assay (a zone of inhibition test of bacterial growth and colony-forming efficiency test of transformant Escherichia coli strains that express mammalian catalase gene derived from normal catalase mice (Cs(a)) and catalase-deficient mutant mice (Cs(b))), Ames mutagenicity assay and H(2)O(2) generation assay. Lawsone generated H(2)O(2) slightly in phosphate buffer system and was not mutagenic in Ames assay using TA 98, TA 100 and TA 102, both in the absence and presence of metabolic activation. Lawsone exposure inhibited the growth of both Cs(a) and Cs(b) strains in a dose-dependent manner. Mean zone diameter for Cs(a) was 9.75+/-0.96 mm and 12.75+/-1.5 mm for Cs(b). Natural henna leaves did not show toxic effects, whereas two out of four samples of marketed henna products were shown toxicity effects. Catalase abolished zone of inhibition (ZOI) of marketed henna products, eliminated ZOI of lawsone in a dose-dependent manner and low concentration of exogenous MnSOD and Cu/ZnSOD eliminated the toxicity. Histidine and DTPA, the metal chelator; BHA and low concentration of capsaicin, the inducer of NADH-quinone reductase, effectively protected Cs(a) and Cs(b) against lawsone in this study. We suggest that lawsone cytotoxicity is probably mediated, at least in part, by the release of O(2)(-), H(2)O(2) and OH(-). PMID:17442476

  19. Rapid analysis of the skin irritant p-phenylenediamine (PPD) in henna products using atmospheric solids analysis probe mass spectrometry.

    PubMed

    Chen, Weiyang; Nkosi, Thobile A N; Combrinck, Sandra; Viljoen, Alvaro M; Cartwright-Jones, Catherine

    2016-09-01

    Henna (Lawsonia inermis) is applied to stain keratin, present in hair, skin and fingernails, a red-orange or rust colour. Producers of temporary tattoos mix the aromatic amine compound, para-phenylenediamine (PPD) into natural henna to create 'black henna' that rapidly stains the skin black. However, PPD may cause severe delayed hypersensitivity reactions following skin contact. This study proposes a rapid direct-analysis method to detect and identify PPD using an atmospheric solids analysis probe (ASAP) coupled to a Q-ToF mass spectrometer (MS). Since laborious, multistep methods of analysis to determine PPD are undesirable, due to the instability of the compound in solution, a screening method involving no sample preparation steps was developed. Experiments were carried out to optimise the corona current, sample cone voltage, source temperature, and desolvation gas temperature to determine ideal ASAP-Q-ToF-MS analysing conditions. Eleven of the 109 henna samples, originating from various countries, tested positive for PPD when henna products were screened using ASAP-MS, without any form of sample preparation other than grinding. Ultra-performance liquid chromatography electrospray ionisation-mass spectrometry (UPLC-Q-ToF-MS) was subsequently used to confirm the results from ASAP and to determine the concentrations of PPD in henna products. The allergen was detected in the same eleven samples, with concentrations ranging from 0.05-4.21% (w/w). It can be concluded that the sensitivity of the ASAP-MS technique is sufficient (limit of detection=0.025% w/w) to allow screening of henna samples for the presence of PPD. This relatively new technique can be applied to commercial products without extraction, sample treatment or chromatographic separation. PMID:27243826

  20. Evaluation of Antifungal Efficacy of Ethanolic Crude Lawsone and Listerine Mouthwash in Uncontrolled Diabetics and Denture Wearers - A Randomized Clinical Trial

    PubMed Central

    Chintamaneni, Rajalakshmi; Chennupati, Anuradha; Nahar, Prashant; Chaluvadi, Rattaiah Setty; Vemugunta, Ramakrishna; Prabhat, Meka Venkata Poorna

    2016-01-01

    Introduction Lawsonia Inermis (LI) is a shrub cultivated throughout India. Many in vitro studies have been done on antifungal activity of LI, although none of the studies have been conducted invivo. Aim To evaluate the antifungal efficacy of ethanolic extract of crude lawsone in comparison with listerine mouth wash in known diabetics and wearing dentures. Materials and Methods A total of 60 subjects were taken and randomly divided into two groups of 30 each. Group 1 received crude lawsone mouthwash and Group 2 received listerine mouth wash. Oral rinse technique was performed. Each subject was given distilled water at baseline and Colony Forming Units (CFU) of candidal species was determined. Post therapeutic samples were then collected 1hr and 1week following drug usage and they were further advised to use given mouth washes twice daily with volume of 5ml/rinse for 30 seconds and CFU was evaluated. Results Crude lawsone mouthwash showed superior antifungal activity when compared to listerine mouthwash. On individual comparison of both mouth washes at baseline, 1hr and 1week highly significant results were obtained using inferential statistics. The inter group comparison was done using independent t-test where lawsone was considered to be more effective in reducing CFU, at 1hr and 1week of using the mouth wash (p<0.01). Subjective symptoms like taste and smell were determined by chi square test where good taste was felt for lawsone and olfactory satisfaction was good with listerine (p<0.01). Burning sensation was found to be more with listerine mouth wash. Conclusion The present study revealed superior antifungal activity with ethanolic extract of crude lawsone mouth wash compared with listerine mouthwash. PMID:27504419

  1. Systematic analysis of in vitro photo-cytotoxic activity in extracts from terrestrial plants in Peninsula Malaysia for photodynamic therapy.

    PubMed

    Ong, Cheng Yi; Ling, Sui Kiong; Ali, Rasadah Mat; Chee, Chin Fei; Samah, Zainon Abu; Ho, Anthony Siong Hock; Teo, Soo Hwang; Lee, Hong Boon

    2009-09-01

    One hundred and fifty-five extracts from 93 terrestrial species of plants in Peninsula Malaysia were screened for in vitro photo-cytotoxic activity by means of a cell viability test using a human leukaemia cell-line HL60. These plants which can be classified into 43 plant families are diverse in their type of vegetation and their natural habitat in the wild, and may therefore harbour equally diverse metabolites with potential pharmaceutical properties. Of these, 29 plants, namely three from each of the Clusiaceae, Leguminosae, Rutaceae and Verbenaceae families, two from the Piperaceae family and the remaining 15 are from Acanthaceae, Apocynaceae, Bignoniaceae, Celastraceae, Chrysobalanaceae, Irvingiaceae, Lauraceae, Lythraceae, Malvaceae, Meliaceae, Moraceae, Myristicaceae, Myrsinaceae, Olacaceae and Sapindaceae. Hibiscus cannabinus (Malvaceae), Ficus deltoidea (Moraceae), Maranthes corymbosa (Chrysobalanaceae), Micromelum sp., Micromelum minutum and Citrus hystrix (Rutaceae), Cryptocarya griffithiana (Lauraceae), Litchi chinensis (Sapindaceae), Scorodocarpus bornensis (Olacaceae), Kokoona reflexa (Celastraceae), Irvingia malayana (Irvingiaceae), Knema curtisii (Myristicaceae), Dysoxylum sericeum (Meliaceae), Garcinia atroviridis, Garcinia mangostana and Calophyllum inophyllum (Clusiaceae), Ervatamia hirta (Apocynaceae), Cassia alata, Entada phaseoloides and Leucaena leucocephala (Leguminosae), Oroxylum indicum (Bignoniaceae), Peronema canescens,Vitex pubescens and Premna odorata (Verbenaceae), Piper mucronatum and Piper sp. (Piperaceae), Ardisia crenata (Myrsinaceae), Lawsonia inermis (Lythraceae), Strobilanthes sp. (Acanthaceae) were able to reduce the in vitro cell viability by more than 50% when exposed to 9.6J/cm(2) of a broad spectrum light when tested at a concentration of 20 microg/mL. Six of these active extracts were further fractionated and bio-assayed to yield four photosensitisers, all of which are based on the pheophorbide-a and -b core structures

  2. An assessment of the genotoxicity of 2-hydroxy-1,4-naphthoquinone, the natural dye ingredient of Henna.

    PubMed

    Kirkland, David; Marzin, Daniel

    2003-06-01

    2-Hydroxy-1,4-naphthoquinone (HNQ; Lawsone; CAS 83-72-7) is the principal natural dye ingredient contained in the leaves of Henna (Lawsonia inermis). Published genotoxicity studies on HNQ suggested it was a weak bacterial mutagen for Salmonella typhimurium strain TA98 or was more clearly mutagenic for strain TA 2637, both in the presence of metabolic activation. HNQ was unable to induce sex-linked recessive lethal mutations in Drosophila melanogaster. However, a small increase in micronucleus frequency was reported in the bone marrow of mice at a single mid-range dose level, 24h after intraperitoneal injection. In view of the wide use of Henna hair dyes it was deemed necessary to conduct a thorough investigation, under Good Laboratory Practice conditions, of the genotoxicity of HNQ. HNQ was non-mutagenic in bacterial (Ames test) or mammalian (V79 hprt) assays. It was borderline positive in a mouse lymphoma tk mutation assay and a chromosome aberration test (CHO cells), results that may reflect a similar clastogenic mechanism. Negative in vivo genotoxicity results were noted in the rat hepatocyte in vivo/in vitro UDS test, in peripheral lymphocytes (chromosome aberrations) of rats receiving repeated oral doses of HNQ at the MTD for 28 days, and in mouse and hamster bone marrow chromosome aberration tests. However small, but statistically significant increases in the incidence of bone marrow micronuclei were observed in two out of five tests at 72 h after dosing, but not at 24 or 48 h. There was evidence of haematotoxicity at 72 h, which may have been enhanced by the vehicle (DMSO) used in the positive tests. As erythropoiesis and administration of haematotoxic agents are known to induce small increases in the frequency of bone marrow micronuclei, typically at delayed sampling times, the data suggest that the positive 72 h response produced by HNQ is consistent with stimulation of haematopoiesis subsequent to haematological toxicity of HNQ, and not due to a DNA

  3. An assessment of the genotoxicity of 2-hydroxy-1,4-naphthoquinone, the natural dye ingredient of Henna.

    PubMed

    Kirkland, David; Marzin, Daniel

    2003-06-01

    2-Hydroxy-1,4-naphthoquinone (HNQ; Lawsone; CAS 83-72-7) is the principal natural dye ingredient contained in the leaves of Henna (Lawsonia inermis). Published genotoxicity studies on HNQ suggested it was a weak bacterial mutagen for Salmonella typhimurium strain TA98 or was more clearly mutagenic for strain TA 2637, both in the presence of metabolic activation. HNQ was unable to induce sex-linked recessive lethal mutations in Drosophila melanogaster. However, a small increase in micronucleus frequency was reported in the bone marrow of mice at a single mid-range dose level, 24h after intraperitoneal injection. In view of the wide use of Henna hair dyes it was deemed necessary to conduct a thorough investigation, under Good Laboratory Practice conditions, of the genotoxicity of HNQ. HNQ was non-mutagenic in bacterial (Ames test) or mammalian (V79 hprt) assays. It was borderline positive in a mouse lymphoma tk mutation assay and a chromosome aberration test (CHO cells), results that may reflect a similar clastogenic mechanism. Negative in vivo genotoxicity results were noted in the rat hepatocyte in vivo/in vitro UDS test, in peripheral lymphocytes (chromosome aberrations) of rats receiving repeated oral doses of HNQ at the MTD for 28 days, and in mouse and hamster bone marrow chromosome aberration tests. However small, but statistically significant increases in the incidence of bone marrow micronuclei were observed in two out of five tests at 72 h after dosing, but not at 24 or 48 h. There was evidence of haematotoxicity at 72 h, which may have been enhanced by the vehicle (DMSO) used in the positive tests. As erythropoiesis and administration of haematotoxic agents are known to induce small increases in the frequency of bone marrow micronuclei, typically at delayed sampling times, the data suggest that the positive 72 h response produced by HNQ is consistent with stimulation of haematopoiesis subsequent to haematological toxicity of HNQ, and not due to a DNA

  4. Global analysis of gene expression dynamics within the marine microbial community during the VAHINE mesocosm experiment in the southwest Pacific

    NASA Astrophysics Data System (ADS)

    Pfreundt, Ulrike; Spungin, Dina; Bonnet, Sophie; Berman-Frank, Ilana; Hess, Wolfgang R.

    2016-07-01

    phosphatase was expressed prominently only between days 12 and 23 in different organisms, suggesting that the microbial community was not limited by phosphate, even before the fertilization on day 4, whereas the post-fertilization community was. We observed high expression of the Synechococcus sqdB gene, only transiently lowered following phosphate fertilization. SqdB encodes UDP-sulfoquinovose synthase, possibly enabling marine picocyanobacteria to minimize their phosphorus requirements by substitution of phospholipids with sulphur-containing glycerolipids. This result suggests a link between sqdB expression and phosphate availability in situ. Gene expression of diazotrophic cyanobacteria was mainly attributed to Trichodesmium and Richelia intracellularis (diatom-diazotroph association) in the Nouméa lagoon and initially in M1. UCYN-A (Candidatus Atelocyanobacterium) transcripts were the third most abundant and declined both inside and outside after day 4, consistent with 16S- and nifH-based analyses. Transcripts related to the Epithemia turgida endosymbiont and Cyanothece ATCC 51142 increased during the second half of the experiment.

  5. Switching between hydrophobic and wettable conditions in soil: experiments to assess the influence of cracks, roots and subsurface drainage impedance

    NASA Astrophysics Data System (ADS)

    Urbanek, E.; Walsh, R. P. D.; Shakesby, R. A.

    2012-04-01

    Although much is known about soil hydrophobicity, assessments of the overall hydrological and erosional significance of the soil property in areas affected by it are greatly hampered by a lack of knowledge on switching between hydrophobic and hydrophilic states. This arises mainly because of (1) the destructive nature of methods of assessing hydrophobicity, (2) its often high local spatial variability and (3) difficulties of relating hydrophobicity results to meaningful soil moisture values. Also, very little is known about the influence which cracks and holes through hydrophobic soil and the presence or absence of subsurface impeding layers have on the 3D pattern and speed of hydrophobicity change during wetting and drying cycles. These issues form the focus of the present paper, which was carried out as part of the EU DESIRE Project. A laboratory experimental approach was adopted. Three different soils of equal initial hydrophibicity class when dry (18 % MED), but of contrasting texture and total carbon content, were investigated: (1) from the scrub-covered (dominated by Erica umbellata, Calluna vulgaris and Pterospartum tridentatum) Vale Torto catchment in Gois municipality, central Portugal (an area where the impacts of prescribed fire were being assessed); (2) soil around a Chamaecyparis lawsonia tree in South Wales; and (3) a vegetated coastal sand-dune location at Nicholaston, Gower Peninsula, South Wales. For the experiments, 106 samples of sieved (< 2 mm), dried soil were placed to a depth of 10 cm in standardized transparent pots (16.5 cm high, top diameter 16 cm, basal diameter 11 cm). Equal numbers of samples were prepared with either (i) five simulated holes, (ii) two simulated linear cracks (in both cases extending downwards to the sample base) and (iii) control soil samples without cracks or holes). Samples were also either (i) sealed at the base to create subsurface impeded drainage or (ii) provided with unimpeded basal drainage by insertion of