Future Benefits of Microorganism on Leather Defects in The Industrial Production of Protease

Article, pickle, and wet blue leather defects used for this study were fromthe Balai Besar Karet college, Kulit dan Plastik, D.I Yogyakarta (BBKKP YK), Indonesia. Meanwhile, the microorganisms in leather defects were grown in vitro at A-minimal mineral (MM) and B-lowest (1/200 v/v) nutrient media. A nitrogen source of 2% Sigma-Aldrich bovine gelatine was added to each medium. Furthermore, 1cm2 of each leather defect was sliced and immersed into the in vitro media for 7 days in an open-air rotary incubation with ambient temperature at 28° C to 30° C. The first or conventional method was the rubbing of ose cotton into the solid media, while the second isolation method was the centrifugationof liquid growth medium at 15.1G for 20 minutes. Moreover, the four microbial isolates were fromglossy yellow colonies A and B as wel as white colonies. These colonies were incubated at 38° C and the four microbes produce proteases after growing for at least 7days in liquid media and 24 hours less in solid media. The protease test produced gases on the pickle leather defects using a test tubeglass of 0.8cmdiameter and 15cmlong. Therefore, the chemical tanning process on leather defects creates a unique ecosystem of microorganisms as collagen proteins change and become thekeyto their growth.

Quantitation of viableCoxiella burnetiiin milk products using a liquid medium-based MPN-PCR assay

This Technical Research Communication describes a new method by which thermally treatedCoxiellain milk products may be grown in a liquid growth medium and quantitated using an MPN-PCR assay.Coxiellais generally not used in studies on thermal and non-thermal processing of milk due to the need for specialized and highly laborious techniques such as animal assays and tissue culture for determining viability. Recently, a liquid growth medium (ACCM-2) and modified atmosphere were used to growCoxiellafrom pure cultures, infected mouse tissues, and clinical samples, however, the ability to growCoxiellafrom a food such as milk has not been shown. The potential ability to enrichCoxielladirectly from contaminated milk presents a new avenue for conducting pasteurization research in which the viability of heat-treated or injured cultures could be more easily determined through direct enrichment ofCoxiellain ACCM-2. ACCM-2 medium allowed enrichment ofCoxiellafrom bovine whole milk and cream, whole goat, and whole camel milks but not whole water buffalo milk. Enrichment was possible from whole bovine milk containing as few as 6Coxiellage/ml of milk. The applicability of this ACCM-2 enrichment method was shown when using an MPN-PCR assay to quantitate the number of viableCoxiellaremaining in whole bovine milk after 64 °C thermal treatment for up to 10 min.

Experimental micropedology—A technique for investigating soil carbonate biogenesis along a desert-grassland-forest transect, New Mexico, USA .

Manipulative experiments—characterized by the comparing treatments to controls—are widespread in scientific investigations. This study uses experimental micropedology to investigate whether soil microbes precipitate carbonate if a liquid growth-medium is applied to soil in situ. This was undertaken using apparatuses designed to (1) obtain micromorphological images of biogenic carbonate on microscope slides, (2) to quantify carbonate formation in fiberglass cloths, and (3) to measure associated carbon-isotope fractionations. The apparatuses were buried and harvested at monthly intervals from December 2010 to June 2011. The study was conducted along an ecological transect in New Mexico, USA, at three sites: a low-elevation desert (C3 shrubs), an intermediate-elevation steppe (C4 grasses), and a high-elevation forest (C3 conifers). In addition to comparing bioclimatic zones, the effect of parent material was also tested using paired limestone and igneous soils at each site. Microscope slides were analyzed with binocular, petrographic, and scanning electron microscopy equipped with an x-ray microanalyser (EDS), and the fiberglass traps were analyzed with x-ray diffraction and a mass spectrometer for carbon concentrations and isotope ratios. Naturally occurring calcified microbes were found at each site in the form of calcified hyphae, needle fiber, and calcified root hairs, with the exception of the forest site on igneous parent material. Liquid growth medium induced microbial calcification regardless of whether the vegetation was desert shrubs, grassland, or forest, and regardless of whether the parent material was igneous or limestone. Thus, the ability of soil microorganisms to biomineralize carbonate when supplied with liquid growth medium in situ is a phenomenon that crosses biomes and is not limited to microbes endemic to either limestone or igneous parent material.

Growth Characteristics of Bartonella henselae in a Novel Liquid Medium: Primary Isolation, Growth-Phase-Dependent Phage Induction, and Metabolic Studies

ABSTRACT Bartonella henselae is a zoonotic pathogen that usually causes a self-limiting infection in immunocompetent individuals but often causes potentially life-threatening infections, such as bacillary angiomatosis, in immunocompromised patients. Both diagnosis of infection and research into the molecular mechanisms of pathogenesis have been hindered by the absence of a suitable liquid growth medium. It has been difficult to isolate B. henselae directly from the blood of infected humans or animals or to grow the bacteria in liquid culture media under laboratory conditions. Therefore, we have developed a liquid growth medium that supports reproducible in vitro growth (3-h doubling time and a growth yield of approximately 5 × 108 CFU/ml) and permits the isolation of B. henselae from the blood of infected cats. During the development of this medium, we observed that B. henselae did not derive carbon and energy from the catabolism of glucose, which is consistent with genome nucleotide sequence data suggesting an incomplete glycolytic pathway. Of interest, B. henselae depleted amino acids from the culture medium and accumulated ammonia in the medium, an indicator of amino acid catabolism. Analysis of the culture medium throughout the growth cycle revealed that oxygen was consumed and carbon dioxide was generated, suggesting that amino acids were catabolized in a tricarboxylic acid (TCA) cycle-dependent mechanism. Additionally, phage particles were detected in the culture supernatants of stationary-phase B. henselae, but not in mid-logarithmic-phase culture supernatants. Enzymatic assays of whole-cell lysates revealed that B. henselae has a complete TCA cycle. Taken together, these data suggest B. henselae may catabolize amino acids but not glucose to derive carbon and energy from its host. Furthermore, the newly developed culture medium should improve isolation of B. henselae and basic research into the pathogenesis of the bacterium.

Complement fixation antibody test for human nocardiosis

Complement-fixing antibody was detected in 13 of 16 patients with histological and/or culture evidence of infection with Nocardia species. The antigen used was a filtrate of soluble substances secreted into liquid growth medium by cultures of N. asteroides. Apparent false positives reactions were found in three of three patients with leprosy and two of five patients with tuberculosis--results similar to some previously reported methods. N. asteroides and mycobacteria share antigens. Only the false positives with tuberculosis are considered a diagnostic problem. No reactions were obtained with sera from 26 patients with other infections and 41 unifected individuals. Whereas previous nocardia serodiagnostic methods have a sensitivity of approximately 50%, our overall sensitivity (81%) compares favorably and included 9 of 11 positive tests in immunocompromised nocardiosis patients (a source of false negative reactions with previous methods).