Invasive ability of Escherichia coli O18 isolated from swine neonatal diarrhea

Neonatal diarrhea occurred at two swine breeding farms in Hokkaido. Ten piglets aged 2 to 4 days were examined. Grossly, significant changes were confined to the small intestine. The mucous membrane was muddy and thickened. The intraluminal contents from the jejunum to the colon were liquid and yellow. In the small intestine, numerous Gram-negative bacilli preferentially adhered to the apex of villi. The mucosa was erosive with villous atrophy. There were bacilli also in the lamina propria and in the cytoplasm of degenerated enterocytes. Nonhemolytic Escherichia coli strains, belonging to serogroup E. coli O18 and possessing K88 fimbriae, were isolated from the small intestine. They could not be classified into any of the diarrheagenic E. coli groups because of the absence of genes of LT, STh, STp, VT1, VT2, eae, invE, and ipaH. After inoculation of the isolates on HEp-2 cells, some bacilli were engulfed by cytoplasmic projections resembling membrane ruffles and subsequently were localized in cytoplasmic vacuoles or free in the cytoplasm. These findings support the view that the present E. coli O18 is a new invasive strain enteropathogenic to piglets.     

Ulcerative keratitis in thoroughbred racehorses in Japan from 1997 to 2008

Objective To describe the number of cases, etiologies, healing times, and results of microbial culture and susceptibility testing of corneal ulcers in thoroughbred racehorses in Japan. Procedure Retrospective study of the medical records of racehorses belonging to the Japan Racing Association (JRA) from 1997 to 2008. Results A total of 2846 horses suffered ulcerative keratitis. These horses accounted for 90.5% of all the corneal problems and 54.9% of the entire number of horses with ocular diseases. Running in races was the cause in 64.3% of the cases. The mean healing time of the ulcers developed in races was 5.5 ± 9.6 days, which was shorter than that of the ulcers unrelated to racing (8.6 ± 11.7 days). In horses presented for examination by the next day after the race, healing was even more rapid (4.1 ± 7.5 days) than in horses presented later (12.4 ± 14.7 days). Microbial culture was performed on 74 samples of 58 horses. Forty‐four bacterial and four fungal organisms were isolated from 35 samples of 29 horses. Pseudomonas aeruginosa (10) and Streptococcus equi subspecies zooepidemicus (5) were the bacteria most frequently isolated. There was no tendency of increasing antibiotic resistance of these organisms. Conclusions Ulcerative keratitis is the most frequent corneal and ocular disease of the racehorse in Japan. Careful observation of the eyes after racing is necessary as early diagnosis and treatment of corneal ulcers speeds healing. Many antibiotic have efficacy against the bacterial isolates, however, potent antibiotics should be reserved for the most severe corneal ulcers.     

Dynamics of C natural abundance in wood decomposing fungi and their ecophysiological implications

Factors that affect the δ¹³C values of fungi need to be analyzed for the progress of isotope-based studies of food-chain or organic matter dynamics in soils. To analyze the factors that control δ¹³C values of the fungal body, basidiomycete and ascomycete species were grown on a beechwood substrate (six species) and in glucose medium (nine species), and the δ¹³C value of produced fungal body was compared to that of the carbon source. The ¹³C enrichment (Δδ¹³C) in the fungal aggregates compared to the decomposed wood varied from 1.2 to 6.3‰ among six species. In the glucose substrate experiment, the degree of ¹³C enrichment in the hyphal mat was relatively small and varied from −0.1 to 2.8‰ among nine basidiomycetes species depending on their growth stage. Calculated δ¹³C values of the respired CO₂ were lower than those of the hyphal mat, organic metabolites and the glucose used. The degree of ¹³C enrichment was affected by fungal species, substrate and growth stage. Fungal internal metabolic processes are the plausible mechanism for the observed isotopic discrimination between fungal bodies and substrates. Especially, dark fixation of ambient CO₂ and kinetic isotope fractionation during assimilation and dissimilation reactions could well explain Δδ¹³C dynamics in our experiments. Through the analysis of field Δδ¹³C, we could know undisturbed fungal status about starvation, aeration and type of decomposition.     

Reversible conformational change in beta-lactoglobulin A modified with N-ethylmaleimide and resistance to molecular aggregation on heating

beta-Lactoglobulin A (beta LG A) modified with N-ethylmaleimide (NEM-beta LG A) was purified by ion exchange chromatography, and modification of beta LG A by NEM was confirmed by time of flight mass spectrometry and 5,5'-dithiobis(2-nitrobenzoic acid) methods. The fluorescent spectrum of NEM-beta LG A was slightly different from that of native beta LG A. NEM-beta LG A gave no polymerization after heating at 80 degrees C and pH 7.5, as shown by polyacrylamide gel electrophoresis. Conformational change of NEM-beta LG A was observed at 80 degrees C by ultraviolet differential spectra, whereas after cooling it recovered to its original state as before heating, indicating apparent reversible thermal denaturation. Native beta LG A is resistant to pepsin hydrolysis, whereas heated beta LG A was easily hydrolyzed by pepsin. NEM-beta LG A before heating was also resistant to pepsin hydrolysis, and after heating NEM-beta LG A was still resistant to pepsin hydrolysis. These results indicate that NEM-beta LG A maintained a conformation similar to its native form even after heating. Addition of 0.2 M NaCl to the beta LG A heated under salt-free condition induced polymerization of heated beta LG A molecules, but not that of heated NEM-beta LG A. This seemed to indicate that the formation of inter- or intramolecular disulfide linkage made the heat-induced conformational change of beta LG A irreversible.     

Low molecular weight compounds responsible for savory taste of Indonesian soy sauce

Indonesian soy sauce is made using only soybeans as the nitrogenous source. Moromi obtained from fermentation of yellow soybeans using Aspergillus sojae as the starter was investigated. The fraction with molecular weights of less than 500 Da obtained by stepwise ultrafiltration was then fractionated by several chromatographic procedures, including gel filtration chromatography and RP-HPLC. Several chemical analyses, CE profiles, and taste profiles were performed to obtain the most intense umami fraction. The main components eliciting or enhancing the umami taste present in the fraction were purified and identified by protein sequencing, ESI-MS, and (1)H NMR at 400 MHz. Besides free l-glutamic acid and aspartic acid, free aromatic amino acids such as l-phenylalanine and l-tyrosine may also play an important role in impressing savory or umami taste of Indonesian soy sauce at their subthreshold concentrations and in the presence of salt and free acidic amino acids. This is reported as a new phenomenon of the so-called bitter amino acids.     

Synergetic effects of cell immobilization in polyurethane foam and use of thermotolerant strain on degradation of mixed hydrocarbon substrate by Prototheca zopfii

ABSTRACT:   The synergetic effects of an immobilized cell system in an oleophilic polyurethane foam (PUF) and use of a thermotolerant strain on degradation of hydrocarbons by colorless green microalga Prototheca zopfii are reported. Two strains of P. zopfii , i.e. thermotolerant RND16 and non-thermotolerant ATCC30253, were immobilized in PUF to compare their abilities to biodegrade a mixed hydrocarbon substrate (MHS) containing aliphatic and polycyclic aromatic hydrocarbons (PAHs), as a function of temperature. The thermotolerant strain RND16 degraded MHS at 35°C, while ATCC30253 did not degrade hydrocarbons at temperatures higher than 30°C. Immobilization of P. zopfii in PUF resulted in shortened lag for growth-associated biodegradation of n -alkanes in MHS, the effect of which was most significant in cultures of RND16 at 25°C. Nevertheless, the decrease in the amount of degraded PAHs was caused by PUF immobilization and the level of this decrease was marked in the cultures of RND16, in which rate and extent of n -alkane degradation were higher than for ATCC30253.     

Acute myeloid leukemia with mastocytic and megakaryocytic differentiation in a calf

A diagnosis of acute myeloid leukemia was made in a 10-month-old Holstein female calf. The leukemia was macroscopically characterized by great enlargement of the spleen and moderate enlargement of some lymph nodes. Histochemical and immunohistochemical examination disclosed the presence of neoplastic cells either containing metachromatic and tryptase-positive granules or expressing factor VIII-related antigen. The granules, which were positive for naphthol AS-D chloroacetate esterase and did not have particulate contents, were distinct from those of basophilic leukemia cells. This leukemia was thought to be derived from a common myeloid progenitor capable of giving rise to megakaryocyte-erythrocyte progenitors and granulocyte-monocyte progenitors with the ability to differentiate into mast cells.     

Seed-coat anatomy and proanthocyanidins contribute to the dormancy of Rubus seed

Rubus seed has a deep double dormancy that restricts germination due to seed coat structure and chemical composition. Improved germination of diverse Rubus species required for breeding improved blackberry and raspberry cultivars is partly dependent on the seed coat structure. This study evaluated the seed coat structure of three species with thin (R. hoffmeisterianus Kunth & C. D. Bouché), medium (R. occidentalis L.) and thick (R. caesius L.) seed coats. The three species exhibited distinctive seed-coat cell composition. The very thin testa (0.086 mm) of R. hoffmeisterianus had little exotesta (surface) reticulation; with the meso- and endotesta composed of sclereids of homogenous shape and size. R. occidentalis had a thick testa (0.175 mm) and a highly reticulate exotesta; the meso- and endotesta were composed of several diverse types of sclereids. R. caesius had the thickest seed coat (0.185 mm) but only moderate exotesta reticulation; the meso- and endotesta were composed of large, irregular, loosely arranged sclereids. R. occidentalis, a medium size seed, was the most heavily lignified with seed-coat thickness similar to R. caesius, the largest seed. Proanthocyanidins (PAs) from dry seed of six Rubus species were extracted and quantified by high performance liquid chromatography. R. hoffmeisterianus, a thin only slightly hard seed, had half the PA (0.45 μg/seed) of R. occidentalis with a thick, extremely-hard seed coat and diverse sclereids (1.07 μg/seed). PA content and sclereid composition both appear contribute to seed coat hardness and resulting seed dormancy. The effectiveness of sulfuric acid for Rubus seed scarification is likely due to degradation of PAs in the testa.