Filamentous fungi and media for cellulase production in solid state cultures

Cellulase production was evaluated in two reference strains (T. reesei Rut-C30 and T. reesei QM9414), two strains isolated from a sugarcane cultivation area (Trichoderma sp. IPT778 and T. harzianum rifai IPT821) and one strain isolated in a program for biodiversity preservation in São Paulo state (Myceliophthora thermophila M77). Solid state cultures were performed using sugarcane bagasse (C), wheat bran (W) and/or soybean bran (S). The highest FPA was 10.6 U/gdm for M77 in SC (10:90) at 80% moisture, which was 4.4 times higher than production in pure W. C was a strong inducer of cellulase production, given that the production level of 6.1 U/gdm in WC (40:60) was 2.5 times higher than in pure W for strain M77; T. reesei Rut-C30 did not respond as strongly with about 1.6-fold surplus production. S advantageously replaced W, as the surplus production on SC (20:80) was 2.3 times relative to WC (20:80) for M77.

Lactobacillus delbrueckii UFV-H2b20 induces type 1 cytokine production by mouse cells in vitro and in vivo

Lactobacillus delbrueckii UFV-H2b20 has been shown to increase clearance of bacteria injected into the blood of germ-free mice. Moreover, it induces the production of type 1 cytokines by human peripheral mononuclear cells. The objective of the present study was to investigate the production of inflammatory cytokines [interleukin-12 (IL-12 p40), tumor necrosis factor-α (TNF-α), and interferon-γ (IFN-γ)] triggered in vitro by live, heat-killed or lysozyme-treated L. delbrueckii UFV-H2b20 and in vivo by a live preparation. Germ-free, L. delbrueckii-monoassociated and lipopolysaccharide (LPS)-resistant C3H/HeJ mice were used as experimental models. UFV-H2b20 induced the production of IL-12 p40 and TNF-α by peritoneal cells and IFN-γ by spleen cells from germ-free or monoassociated Swiss/NIH mice and LPS-hyporesponsive mice (around 40 ng/mL for IL-12 p40, 200 pg/mL for TNF-α and 10 ng/mL for IFN-γ). Heat treatment of L. delbrueckii did not affect the production of these cytokines. Lysozyme treatment decreased IL-12 p40 production by peritoneal cells from C3H/HeJ mice, but did not affect TNF-α production by these cells or IFN-γ production by spleen cells from the same mouse strain. TNF-α production by peritoneal cells from Swiss/NIH L. delbrueckii-monoassociated mice was inhibited by lysozyme treatment. When testing IL-12 p40 and IFN-γ levels in sera from germ-free or monoassociated Swiss/NIH mice systemically challenged with Escherichia coli we observed that IL-12 p40 was produced at marginally higher levels by monoassociated mice than by germ-free mice (40 vs 60 ng/mL), but IFN-γ was produced earlier and at higher levels by monoassociated mice (monoassociated 4 and 14 ng/mL 4 and 8 h after infection, germfree 0 and 7.5 ng/mL at the same times). These results show that L. delbrueckii UFV-H2b20 stimulates the production of type 1 cytokines in vitro and in vivo, therefore suggesting...

Immunochemotherapy in American cutaneous leishmaniasis: immunological aspects before and after treatment

In this study, we evaluated the immune response of patients suffering from cutaneous leishmaniasis treated with two distinct protocols. One group was treated with conventional chemotherapy using pentavalent antimonium salts and the other with immunochemotherapy where a vaccine against cutaneous leishmaniasis was combined with the antimonium salt. Our results show that, although no differences were observed in the necessary time for complete healing of the lesions between the two treatments, peripheral blood mononuclear cells from patients treated by chemotherapy showed smaller lymphoproliferative responses at the end of the treatment than those from patients in the immunochemotherapy group. Furthermore, IFN-gamma production was also different between the two groups. While cells from patients in the chemotherapy group produced more IFN-gamma at the end of treatment, a significant decrease in this cytokine production was associated with healing in the immunochemotherapy group. In addition, IL-10 production was also less intense in this latter group. Finally, an increase in CD8+ -IFN-gamma producing cells was detected in the chemotherapy group. Together these results point to an alternative treatment protocol where healing can be induced with a decreased production of a potentially toxic cytokine

Influence of dietary protein content on Trypanosoma cruzi infection in germfree and conventional mice

Camundongos isentos de germes (GF) e convencionais (CV) foram alimentados com racoes contendo 4,4, 13,2 ou 26,4 por cento de proteina nao ganharam peso durante quatro semanas, enquanto a racao deficiente em proteina nao afetou o crescimento dos camundongos GF. Apos quatro semanas nessas racoes, os camundongos foram inoculados com 5x10 elevado ao cubo tripomastigotos de Trypanosoma cruzi. A deficiencia em proteina afetou menos os camundongos GF do que os CV, segundo os seguintes parametros: ganho em peso, hemoglobina, niveis em proteina e albumina no plasma e conteudos em agua e proteina na carcassa. A infeccao com T. cruzi produziu um decrescimo significante nos niveis em hemoglobina, na contagem de celulas vermelhas sanguineas e nos conteudos em agua e proteina na carcaca. Este decrescimo foi mais acentuado nos camundongos GF...

Effect of an essential fatty acid deficient diet on experimental infection with Trypanosoma cruzi in germefree and conventional mice

1) To investigate the possible role of essential fatty acid deficiency in host cell/parasite interaction, weanling germefree (GF) and conventional (CV) CFW mice maintained on an essential fatty acid-deficient (-) or a control (+) diet for 110 days were infected with Trypanosoma cruzi. 2) Blood parasitemia indicated that the disease was milder in the animals maintained on the essential fatty acid-deficient diet than in the GF and CV controls (maximum parasitemia: GF+33,300,GF-26,200,CV+17,100 and CV-6,400 trypomastigotes?ml blood). 3) Survival 30 days a after infection was 12% for GF+,28% for GF-,37% for CV+ and 65% for CV- mice. 4) Linoleic and arachidonic acid levels were significantly lower in animals kept on the essential fatty acid-deficient diet (GF-:28.0 ñ 9.3, 23.4 ñ 8.6; CV-:37.6 ñ 5.8, 19.9 ñ 3.6) than in controls (GF+: 164 ñ 48.8, 162.6 ñ 45.8; CV+: 147.1 ñ 26.5, 107.5 ñ 23.6) confirming the deficiency. Before the infection, succinic dehydrogenase levels were higher in liver of all CV mice *4.52 ug phosphate/mg fresh tissue) than in GF mice (0.84 ug phosphate/mg fresh tissue) whereas the opposite was true for 5'-nucleotidase levels in brain and liver, respectively (GF:2.84 and 3.18 ug phosphate/mg fresh tissue: CV:1.25 and 1.54 ug phosphate?mg fresh tissue). The disease was milder in deficient than in control animals in both the GF and CV groups on the basis of parasitemia and survival, indicating that fatty acid-deficient mice are partially protected against Chagas'disease. The mechanism underlying this phenomen requires further investigation

Immune responses of germfree mice to experimental infection with trypanosoma cruzi

1. The immune responses to Trypanosoma cruzi infection of germfree mice were compared to the reponses of infected conventional mice. Two groups (40 animals in each group) of 2-month old female CFW germfree and vonventional mice were used. The IgM and IgG which bound to the surface of T. cruzi epimastigotes determined by ELISA were significantly lower in germfree than in conventional mice (1/3 and 1/5 for IgM and IgG, respectively). 2. After infection there was a three-fold increase in the circulating levels of these immunoglobulins in germfree but not in conventional mice. twenty-one days after T. cruzi inoculation, both IgG and IgM levels were similar in germfree and conventional animals. 3. Footpad swelling after T. cruzi-antigen inoculation was initially four-fold more intense in germfree than in conventional mice. 4. These results suggest that the reduced humoral immune response of germfree mice during ythe initiation of experimental Chagas' disease may be responsible for the more severe parasitism when compared to conventional mice