Supplementation with humic substances affects the innate immunity in layer hens in posfasting phase / El suplemento de sustancias húmicas afecta la inmunidad innata en gallinas ponedoras en fase posmuda

Objective. Asses the effect of supplementation with Humic substances (HS) over some innate immunity parameters (serum bactericidal activity, phagocytosis, bacterial agglutination, respiratory burst and lisozyme activity) in phase after fasting of layer hens. Materials and methods. 120 posfasting phase Hy Line Brown layer hens were taken which were distributed into four groups: The first and the second were supplemented with 0.1 and 0.2% of HS, respectively. The third group was supplemented with 0.25 mg/kg on levamisole hydrochloride and fourth group have no supplementation; during sixty days period. Blood samples were collected on 8th, 30th and 60th of experiment day. Results. The phagocytic index and respiratory burst increased significantly at day 30th in HS supplemented groups. Alike, serum bactericidal activity and lisozyme activity improved on 8 th day, nevertheless, changes were no evident latter. The bacterial agglutination was high in supplemented groups evaluated at everyone times. Conclusions. Results showed that HS behave as immunostimulant in the early phase after fasting layer hens.

Objetivos. Evaluar el efecto de las sustancias húmicas (SH) sobre algunos parámetros de la inmunidad innata (actividad bactericida del suero, fagocitosis, aglutinación bacteriana, explosión respiratoria y actividad de la lisozima) en la fase posmuda de gallinas ponedoras. Materiales y métodos. Se utilizaron 120 gallinas ponedoras Hy Line Brown en la fase de posmuda, las cuales fueron divididas en cuatro grupos: Los dos primeros fueron suplementados con 0.1 y 0.2% de SH respectivamente, el tercer grupo fue suplementado con 0.25 mg/kg de Clorhidrato de levamisol y el cuarto grupo control sin suplemento; durante un período de 60 días. Las muestras sanguíneas se tomaron los días 8, 30 y 60 del experimento. Resultados. El Índice fagocítico y la explosión respiratoria se incrementaron significativamente a partir del día 30 de suplementación con SH. De la misma manera, la actividad bactericida del suero y la actividad de la lisozima aumentaron al día 8; no obstante no se evidenciaron cambios posteriores. La aglutinación bacteriana fue significativamente mayor en los grupos suplementados en todos los tiempos evaluados. Conclusiones. Los resultados demuestran que las SH se comportan como agentes inmunoestimulantes en la fase temprana de la posmuda en gallinas ponedoras Hy Line Brown.

Probiotic features of two oral Lactobacillus isolates

In this study, we checked lactobacilli strains of human origin for their potential as probiotic. Samples were collected from oral mucosa of 16 healthy individuals, out of which twenty isolates were obtained and two of them were selected and identified as Lactobacillus plantarum (G1) and L. casei (G3). Both isolates exhibited antagonistic action towards pathogenic microorganisms such as Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Salmonella abony, and Clostridium sporogenes, but not on the growth of Candida albicans. The bacteriocin activity against Staphylococcus aureus ATCC 6358-P was shown only by L. plantarum G1. Moreover, the isolates G1 and G3 showed good viability in the acid gastric environment and in the gut environment containing bovine bile salts. The viability of G1 and G3 isolates in the gastrointestinal tract, and the adhesion to the intestinal mucosa were also confirmed in vivo. The biochemical tests of blood samples revealed lower levels of serum triglycerides and cholesterol, as well as reduced activity of alkaline phosphatase in all lactobacilli-treated Wistar rats, compared to control ones. No toxicity for NMRI Ham mice was observed. According to our experimental results, these findings imply that L. plantarum G1 and L. casei G3 could be characterized as potential probiotics.

Difusão de microrganismos, inibição do crescimento e atividade biocida do cianoacrilato super bonder / Diffusion, growth Inhibition and biocide activity of the super bonder cyanoacrylate

Os objetivos do presente trabalho foram avaliar se o Super Bonder" apresenta algum risco de veiculação de microorganismos em humanos e se possui alguma atividade bactericida ou bacteriostática para cocos gram-positivo. Foi testada a esterilidade de dez tubas do adesivo nos meios de cultivo BHI, agar sangue, agar Sabouraud com cloranfenicol e Mycosel. A provável ação bactericida ou bacteriostática do adesivo Super Bonder"', in vitro, foi verificada utilizando cepas de Staphylococcus aureus e staphylococcus epidermidir, semeadas em meio de cultura agar sangue e agar Mueller-Hinton. Nao foi observado crescimento de microrganismos nos meios de cultura contendo somente gotas de Super Bonder"'. No agar sangue, observou-se ausência de beta-hemólise e de crescimento bacteriano nas áreas de sobreposição do adesivo. Em agarMueller-Hinton, houve rarefação de colônias ao redor das gotas do cianoacrilato.Os dados deste trabalho mostram que a cola de cianoacrilato Super Bonder"' parece ser livre de contaminação por microrganismos e apresenta possfvel atividade bacteriostatica in vitro.

The purposes of the present study were to verify the human's risk of the Super Bondet microorganisms' diffusion and the probably biocide or bacteriostatic adhesive's activity for gram-positive cocci. The sterility of ten tubes of the adhesive was tested in BHI culture tube and blood-agar, agar-Sabouraud with cloranfenicol and Mycosel culture plates. The biocide or bacteriostatic activity of the Super Bondet adhesive, in vitro, was verified using cultures of Staphylococcus aureus and staphylococcus epidermidis,seeded on blood-agar and agar-Mueller-Hinton culture plates. No microorganisms' growth was observed in the culture plates containing only drops of the Super Bondet. In agar-Mueller-Hinton, zones with reduced number of colonies were noticed around the cyanoacrylate's drops. In blood-agar, absence of haemolysis-beta and bacterial growth were observed in the covered areas of the adhesive. According to the results of this investigation, the Super Bondet cyanoacrylate is free of microorganisms' diffusion and probably inhibits in vitro the bacterial growth.

[Ex-vivo assessment of meningococcal resistance to natural immunity: serum bactericidal assays versus whole- blood assay]

Literature review indicates that antibiotic resistance of bacteria may be associated with the resistance to host natural immunity. In the previous study we found that: all meningococcal isolates of meningitis cases are less sensitive to typical anti-meningococcal antibiotics. In present study, the association of the above characters was analyzed in an ex-vivo "case-control" study. A randomized controlled trial compared effect two dietary treatments [low glycemic index and low fat] in 46 adults, ages 18-55 years old, BMI >27,who proceed to motahary clinic in shiraz, Iran. Body weight, BMI, WHR [waist/hip ratio], fast and post-prandial Plasma lipid profile [Triacylglycerol, total Cholesterol, LDL-C, HDL-C concentration] level of obese women measured at the beginning and at the end 6 weeks. The resistance of both groups against selected sera were similar in the ratio 1:2- by SIC and SBC methods. The growth of "test" and "control" groups in the pooled sera were stopped at the ratios 1:512 and 1:1024 respectively. In SB A the population of "test" and "control" groups decreased in parallel to each other. However, the invasive [test] meningococcal strains established slower reduction in WBA. Studies on the correlation between antibiotic resistance of bacteria and the resistance to the natural immunity have ended with different results. According to our experience, it seems that for ex- vivo investigation of natural immunity against meningococci the WBA is more reliable

Bactericidal activity of GLL-37, a novel derivative of the human antimicrobial peptide LL-37 / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To develop and investigate GLL-37, a substitution analogue of the human antimicrobial peptide LL-37 with anti-enzymatic degradation activity and improved efficacy.</p><p><b>METHODS</b>The bactericidal activities of LL-37 and newly developed GLL-37 against 6 Gram-negative and -positive bacteria were determined by Broth microdilution assays. The minimum inhibitory concentrations of LL-37 and GLL-37 against E.coli ATCC 25922 in different NaCl concentration medium were also detected. Both peptides were co-incubated with elastase, and then analyzed by PAGE electrophoresis and bactericidal activity determination.</p><p><b>RESULT</b>GLL-37 showed a stronger elastase resistance ability than LL-37, and was significantly more effective than LL-37 under high-salt condition.</p><p><b>CONCLUSION</b>The antimicrobial peptide GLL-37 derived form LL-37 has the potential as a new therapeutic agent for bacterial infections.</p>

Distribution of secretory inhibitor of platelet microbicidal protein among urethral isolates with its correlation with prostatitis / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To report the detection in vitro of secretory inhibitor of platelet microbicidal protein (SIPMP) phenotypes of urethral isolates along with a comparison with isolates from patients with or without chronic bacterial prostatitis (CBP).</p><p><b>METHODS</b>Urethral isolates of Staphylococcus spp. (n=4), diphtheroids (n=28), micrococci (n=15), streptococci (n=21), Enterobacteriaceae (n=9) and Enterococcus faecalis (n=19) from patients with or without CBP were tested. SIPMP production was tested by inhibition of platelet microbicidal protein (PMP) bioactivity against Bacillus subtilis and was expressed as percentage of inhibition of PMP bactericidal activity.</p><p><b>RESULTS</b>A significantly higher proportion of CBP-strains (57.78% vs. 16.67%) reduced PMP-induced killing of Bacillus subtilis than non-CBP strains did (P<0.01). SIPMP levels of staphylococci and Enterococcus faecalis from the CBP group were significantly higher than those of the control group.</p><p><b>CONCLUSION</b>These results suggest that SIPMP production is associated with the CBP source. Data from the present study might have significant implications for the understanding of the pathogenesis of CBP.</p>

Virulence factors and drug resistance in Escherichia coli isolated from extraintestinal infections.

PURPOSE: To determine the virulence factors produced by Escherichia coli isolated from extraintestinal infections, to study the drug resistance pattern in E. coli with special reference to extended spectrum beta -lactamase (ESBL) and to evaluate screening methods for ESBL. METHODS: A total of 152 isolates of E. coli from various extraintestinal infections were screened for virulence factors such as haemolysin, surface hydrophobicity, serum resistance and protease. All the isolates were also studied for antibiotic susceptibility pattern using modified Kirby Bauer disk diffusion method. ESBL production was screened by standard disk diffusion method and confirmed using phenotypic confirmatory method. RESULTS: Among 152 isolates, 36 (23.7%) were haemolytic, 42 (27.6%) were hydrophobic, 132 (86.8%) were serum resistant and only four were positive for protease. Multiple virulence factor were observed in 67 (44%) of isolates. Seventy-nine (51.4%) isolates produced ESBL. ESBL producing isolates showed multidrug resistance. There was a significant association ( P E. coli . CONCLUSIONS: The present study shows the expression of virulence factors and multidrug resistance in E. coli isolated from various extraintestinal infections. The study also shows that appropriate methods of detecting drug resistance and ESBL production are required for the judicious use of antibiotics in managing these infections.

In vitro resistance to human platelet microbicidal protein among urethral staphylococcal and enterococcal isolates with its correlation with prostatitis.

The study was carried out to test the in vitro activity of human platelet microbicidal protein (hPMP) on most commonly isolated urethral pathogens and compare the same with clinical isolates from cases of chronic prostatitis (CP). Urethral isolates of Staphylococcus aureus (n=19), coagulase negative staphylococci (n=40) and Enterococcus faecalis (n=16) from patients with or without CP were tested. The hPMP susceptibility of bacterial strains was determined by exposing bacterial cells to serial dilutions of hPMP. A significantly higher proportion of CP-strains of coagulase negative staphylococci (91.3% vs 5.88%) was resistant to hPMP than was that of non-CP strains (P S.aureus studied, 77.8% were considered resistant to the bactericidal action of hPMP. All nine CP-strains of E.faecalis were highly resistant to hPMP. Most non-CP urethral isolates of S.aureus, coagulase negative staphylococci and E.faecalis were susceptible to the bactericidal action of hPMP, while CP isolates of all species were significantly more resistant to hPMP. Data from the present study may have significant implications in understanding the pathogenesis of CP.

Effect of temperature on phagocytic activity of neutrophils.

The effect of temperature on phagocytic activity of neutrophils has been evaluated. Freshly collected heparinised blood samples from young healthy volunteers were incubated with heat killed Staphylococcus aureus at 37 degrees C, 38 degrees C, 39 degrees C and 40 degrees C for 20 minutes. Some of the neutrophils engulfed some heat killed bacteria. Then the blood smears were prepared and stained by Leishman's stain. Engulfed bacteria within the neutrophils stained intensely were observed and counted to find out the phagocytic index and avidity index of the neutrophils. It was found that phagocytic index increased significantly at 38 degrees C and 39 degrees C in comparison to that of at 37 degrees C but did not show significant increase when incubated at 40 degrees C. It seems that contractile elements responsible for the movement of the neutrophils through the formation of pseudopod is more activated at higher temperature (38 degrees C and 39 degrees C) in comparison to that of at normal body temperature (37 degrees C). Temperature higher than 39 degrees C may cause impairment in enzyme function responsible for assembly and disassembly of actin and myosin filaments in the cell causing decreased movement and decreased rate of formation of psudopod resulting in decreased phagocytic activity.

Serum resistance of Escherichia coli strains causing urinary tract infection and diarrhoea in relation to alpha haemolysin production and O type.

A total of 46 alpha-hemolytic and 40 non-hemolytic clinical isolates of Escherichia coli were collected from pediatric patients with urinary tract infection and diarrhoea. Of 39 (84.7%) alpha-hemolytic strains and 27 (67.5%) non-hemolytic strains were resistant to 10% serum and there was no significant difference between urinary and stool isolates. On the contrary when 100% serum was used, 22 (47.8%) of the alpha-hemolytic and 7 (17.5%) of the non-hemolytic strains were resistant (p<0.01). and significantly greater resistance was found in urinary tract infection than from the stool samples (47% versus 24%, p<0.01). Serum resistance was higher in serogroups O6, O18 and O75. Production of alpha-hemolysin was more frequent in serogrops O2, O6, O8, O18 and O75. Thus, the resistance to human serum can determine clinical significance of Escherichia coli from different sources and alpha-hemolysin contributes to the virulence of Escherichia coli in initiation and perpetuation of clinical infection.

Uropathogenic Escherichia coli--a preliminary study.

Urinary isolates of Escherichia coli were studied for presence of haemolysin, adhesins, serum resistance and O serotype prevalence. Of the 144 isolates studied, 72 exhibited hemolysin, 7 were resistant to bactericidal effect of serum and 50 strains showed Mannose resistant Haemagglutination (MRHA). O101,O68,O04 and O25 were the commonest serotypes in this study.

Killing of Escherichia coli strains from clinical specimens in human serum and polymorphonuclear leucocytes.

Hundred Escherichia coli strains were collected from extra-intestinal and intestinal disease for the present study. Of the strains isolated 49 (49%) were serum sensitive and 47 (47%) serum resistant. The remaining 4 (4%) strains showed intermediate sensitivity to the pooled normal human serum (PNHS). Strains isolated from faeces were significantly more sensitive than strains of extra-intestinal origin (P<0.01). Response of Escherichia coli strains to killing by polymorphonuclear leucocytes was seen in 50 isolates (50%). Faecal strains showed significantly more intracellular killing as compared to extra-intestinal strains (P<0.01). Thus, clinical significance of Escherichia coli strains from different sources can be determined by the resistance to the bactericidal effect of human serum and killing in polymorphonuclear leucocytes.

Construction, expression and biological assessment of BPI23-Fcgamma1 recombinant protein prokaryotic expression vector / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To construct pBV-BPI600-Fcgamma1(700) recombinant expression vector, to transform it into Escherichia coli DH5alpha, and to induce the expression of BPI23-Fcgamma1 anti-bacterial recombinant protein.</p><p><b>METHODS</b>Genes coding for BPI23 and Fcgamma1 were amplified by RT-PCR from mRNA extracted from HL-60 cell and normal human leukocytes; recombinant cloning vector and recombinant expression vector were then constructed. pBV-BPI600-Fcgamma1(700) recombinant expression vector was transformed into the competent Escherichia coli DH5alpha and BPI23-Fcgamma1 recombinant protein was expressed by a temperature-induced method.</p><p><b>RESULTS</b>(1) Expected amplified products BPI600hp and Fcgamma1(700bp) were obtained by RT-PCR method. (2) pUC18-BPI180, pUC18-BPI420 and pUC18-Fcgamma1(700) recombinant cloning vector were successfully constructed, and sequences were identical with the reported ones. 3) pBV-BPI600-Fcgamma1(700) recombinant expression vector was successfully constructed, and the enzyme digestion analysis showed an expected result. (4) The expression level of BPI23-Fcgamma1 recombinant protein accounted for 20% of total bacterial proteins. (5) The renatured BPI23-Fcgamma1 recombinant protein showed bacteriocidal activity and biological function of complement fixation, and opsonization.</p><p><b>CONCLUSION</b>pBV-BPI600-Fcgamma1(700) recombinant expression vector was successfully constructed, and BPI23-Fcgamma1 recombinant protein with double biological activity of BPI and IgGFc was expresed in Escherichia coli.</p>

Estandarización y evaluación del ensayo vibriocida modificado / Standardization and evaluation of the modified vibriocidal assay

The vibriocidal antibody test is a reliable and well-documented method to determine bacterial antibodies to Vibrio cholerae 01 antigens. It consists of mixing serum dilutions and a steady quantity of bacteria and supplement to cause cell lysis. Titer is determined by visual observation. In this paper, we implemented a change in the presented method where a pH and glucose indicator was added to the culture medium used to stop the reaction, which allowed a quicker reading by any person who are not very familiar with the carrying out of this test since the colour change in the plaque is quite evident.

IL-2 and IFN-gama, but not IL-4 secretion by peripheral blood mononuclear cells (PBMC) are related to CD4+ T cells and clinical status in brazilian HIV-1-infected subjects

E relatado que a producao de IL-2 e IFN-gama, conhecidas como citoquinas T-auxiliador tipo 1, pelas celulas mononucleares do sangue periferico fica deprimida no decorrer da infeccao HIV. Por outro lado, a producao de IL-4 e IL-10, chamada padrao T-auxiliador tipo 2, aumenta com o avanco da doenca (AIDS). Nesse estudo, foram avaliados 55 individuos infectados pelo HIV-1 em acompanhamento no Ambulatorio da Imunodeficiencias Secundarias do Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo. Celulas mononucleares foram estimulados "in vitro" com fitohemaglutina (PHA) por 24 horas e o sobrenadante foi utilizado para a dosagem de citoquinas atraves de kits de ELISA disponiveis comercialmente. Foi observado que a producao de IFN-y pelos individuos assintomaticos HIV+ esta aumentada quando comparada aos controles nao infectados pelo HIV, enquanto os pacientes com AIDS tiveram producao similar aos controles...

Efecto del levamisol en la actividad microbicida y quimiotaxis en células polimorfonucleares / The effect of levamisol on the functions of chemotaxis and bacterial activity

Antecedentes: la septicemia neonatal puede tener una mortalidad del 20 por ciento en recién nacidos pequeños para su edad gestacional. Esta predisposición a las infecciones se ha explicado por alteraciones inmunológicas. El propósito de este trabajo es conocer el efecto del levamisol en las funciones de quimiotaxis y actividad bactericida de las células polimorfonucleares en recién nacidos pequeños para su edad gestacional. Material y método: se estudiaron 20 recién nacidos de término. Diez eran pequeños para su edad gestacional y los otros 10 tenían peso adecuado. Se midió la actividad microbicida y la quimiotaxis; se hicieron comparaciones entre los grupos con pruebas estadísticas no paraméticas. Resultados: en el grupo de neonatos pequeños para su edad gestacional la actividad microbicida fue semejante a la del grupo testigo, pero la actividad quimiotáctica estuvo disminuida (p < 0.05)

Study on bactericidal effects of ultraviolet rays in vitro

Having studied in vitro the effect of UVR on the two species of microorganisms which are most common as agents of surgical infection, we realized the closer the distance from the irradiation lamp and the longer the irradiation time, the stronger the bactericidal effect of UVR. When UVR is applied for air disinfection within the distance of more than 2 meters from the sunlamp irradiation, it can also reduce significantly the number of microorganisms in the air. It is due to the fact that there is a continuous air exchange between the upper and the lower part of the room. Previous studies and the experiments on petri dishes show that the use of UVR for cleaning infected wounds and preventing infection is an effective, simple and safe measure

Inhibition of phagocytosis of E. coli by B. fragilis group--an in vitro study.

Bacteriodes fragilis group can inhibit the phagocytosis and killing of co-existing aerobes. Forty six strains of B.fragilis group are tested for their ability to resist phagocytosis and inhibit the killing of indicator organism, E. coli by denoting the viable count. Among the species of B. fragilis group, the inhibitory index of the phagocytic system was the highest with B. fragilis followed by B. thetaiotamicron. None of the strains of B. fragilis group were phagocytosed or killed in this system. This property of resisting the phagocytosis and killing of E. coli by B. fragilis group might be the contributing factor towards their prevalence in mixed infections.

Sensitivity to the bactericidal effect of human serum of Proteus strains from clinical specimens.

A total of 257 Proteus strains isolated from urinary tract infection, blood, wound and faeces were studied. Of the strains tested 31 (12 percent) were serum sensitive, 182 (71 percent) were serum resistant and the remaining 44 (17 percent) showed intermediate sensitivity to the pooled normal human serum (PNHS). Strains isolated from adult urines and blood cultures were significantly more sensitive than strains of faecal origin (p < 0.01). No significant difference was seen between strains from faeces and wounds.