Journal: MicrobiologyOpen

Article Title: Recombinant Limosilactobacillus ( Lactobacillus ) delivering nanobodies against Clostridium perfringens NetB and alpha toxin confers potential protection from necrotic enteritis

doi: 10.1002/mbo3.1270

Figure Lengend Snippet: Inhibitory activity of L. reuteri 3632 (a) and L. reuteri 3630 (b) against C. perfringens in an agar overlay assay. The data are representative of three independent experiments

Article Snippet: Fresh C. perfringens (strain CL‐15, Type A, α and β2 toxins) challenge inoculum was prepared every day from a stock culture in fluid thioglycollate broth overnight at 35°C under anaerobic conditions by Microbial Research Inc. C. perfringens was administered by oral gavage at a target dose of approximately 1 × 10 8 CFUs/ml (1 ml/chick) on 17, and 18 days of age.

Techniques: Activity Assay, Overlay Assay

Journal: MicrobiologyOpen

Article Title: Recombinant Limosilactobacillus ( Lactobacillus ) delivering nanobodies against Clostridium perfringens NetB and alpha toxin confers potential protection from necrotic enteritis

doi: 10.1002/mbo3.1270

Figure Lengend Snippet: Neutralization of NetB activity by L. reuteri secreted Nbs. (a) A twofold dilution series of precipitated VHH antibodies (5 µM) was preincubated with recombinant NetB, after which 1% chicken erythrocytes was added. The optical density of 100% hemolysis was obtained by diluting the chicken erythrocytes in distilled water. As a control, chicken erythrocytes were incubated with NetB, but without Nbs was used. This resulted in 100% hemolysis (OD 570 = 0.54). A NetB positive control (NetB in PBS) resulted in a mean OD 570 of 0.52 and a PBS negative control (PBS with no NetB and Nbs) yielded a mean OD 570 of 0.05. As the initial amounts of L. reuteri and E. coli purified Nbs used for the assay were different, normalized OD 570 values are shown. (b) Western blot analysis binding of anti‐NetB Nb to NetB in the culture supernatant from different C. perfringens clinical isolates. 1, Ladder; 2, NetB positive control (5 µg); 3, C. perfringens JP1011 overnight culture supernatant (10 µl); 4, C. perfringens JP1011 overnight culture supernatant, 10× concentrated (10 µl); 5, C. perfringens JP1011 mid‐log culture supernatant (10 µl); 6, C. perfringens JP1011 midleg culture supernatant, 10× concentrated (10 µl); 7, C. perfringens CP1‐1 overnight culture supernatant (10 µl). The data represent the mean ± SD of the results of three independent experiments. Nbs, nanobodies; PBS, phosphate‐buffered saline; VHH, Variable domain of the Heavy chain of Heavy chain

Article Snippet: Fresh C. perfringens (strain CL‐15, Type A, α and β2 toxins) challenge inoculum was prepared every day from a stock culture in fluid thioglycollate broth overnight at 35°C under anaerobic conditions by Microbial Research Inc. C. perfringens was administered by oral gavage at a target dose of approximately 1 × 10 8 CFUs/ml (1 ml/chick) on 17, and 18 days of age.

Techniques: Neutralization, Activity Assay, Recombinant, Control, Incubation, Positive Control, Negative Control, Purification, Western Blot, Binding Assay, Saline

Journal: MicrobiologyOpen

Article Title: Recombinant Limosilactobacillus ( Lactobacillus ) delivering nanobodies against Clostridium perfringens NetB and alpha toxin confers potential protection from necrotic enteritis

doi: 10.1002/mbo3.1270

Figure Lengend Snippet: Efficacy of NE01 and NE06 on reduction of NE‐associated mortality. Efficacy of NE01 and NE06 was evaluated using a dual challenge model using Eimeria maxima and C. perfringens challenge as described in Section . Chickens that died postchallenge phase between 18 (after challenge with C. perfringens ) and 28 days of age were necropsied, cause of death was listed as NE‐related or non‐NE‐related mortality, and % NE mortality was calculated. a p < 0.05; b p < 0.05; c p = 0.15. CFU, colony‐forming unit; NE, necrotic enteritis

Article Snippet: Fresh C. perfringens (strain CL‐15, Type A, α and β2 toxins) challenge inoculum was prepared every day from a stock culture in fluid thioglycollate broth overnight at 35°C under anaerobic conditions by Microbial Research Inc. C. perfringens was administered by oral gavage at a target dose of approximately 1 × 10 8 CFUs/ml (1 ml/chick) on 17, and 18 days of age.

Techniques:

Journal: Microorganisms

Article Title: Preservation, Characterization and Exploitation of Microbial Biodiversity: The Perspective of the Italian Network of Culture Collections

doi: 10.3390/microorganisms7120685

Figure Lengend Snippet: Partners and associated member of the joint research unit MIRRI-IT.

Article Snippet: Consiglio Nazionale delle Ricerche (CNR) 1 , Agro-Food Microbial Culture Collection (ITEM) , Bacteria, Filamentous fungi, Yeasts , Air, Insects, Food, Plants, Soil.

Techniques: Isolation, Bacteria, Virus