Characterization of "Pasteurella" piscicida isolated from white perch and cultivated yellowtail.

The morphological, physiological, and biochemical characteristics of twelve "Pasteurella" piscicida strains isolated from white perch and yellowtail are described and the present uncertain taxonomic status of the organisms is discussed. The organisms isolated were gram-negative rods showing bipolar staining and pleomorphism. No spores or flagella were observed. They were non motile and viscid colonies were formed. Growth was observed in a temperature range of 20 to 30 C and the salinity range of growth was between 0.5% and 4.0%. They were aerobic and facultative anaerobic. Oxidase and catalase were produced. Nitrates were not reduced to nitrites. Lysine and ornithine decarboxylases were not produced but arginine dihydrolase was produced. The egg yolk reaction was positive. Tween 80 and tributyrin were decomposed. Phosphatase was produced. Beta hemolysis was revealed on a medium containing defibrinated blood from chickens or carp but not from mammals. Methyl red and Voges-Proskauer tests were positive, and acetoin was produced from 2,3-butanediol. Glucose was fermented without gas production. Acid was produced from fructose, mannose, galactose, sucrose, maltose, dextrin and glycerol. These organisms differed from all other members of the genus Pasteurella with respect to nitrate reduction, arginine dihydrolase, Voges-Proskauer and methyl red tests. The formation of viscid colonies and inability to grow in a medium without sodium chloride or at 37 C were additional characteristics of these organisms.     

红拟石首鱼海豚链球菌分离、鉴定及致病性研究

2001年9月-12月,浙江舟山部分网箱养殖红拟石首鱼发生了陆续死鱼,发病鱼表现为眼球突出、浑浊,失去方向性,皮肤溃疡等症状。从病鱼的肾脏和肝脏中分离到菌株SO-2和SO-3,对两分离株进行了致病性试验,发现两者对红拟石首鱼、罗非鱼及小白鼠均有致病力,SO-2和SO-3对红拟石首鱼、罗非鱼及小白鼠的LD50分别为4.8×108CFU/尾和1.9×107CFU/尾2、.8×108CFU/尾和8.3×107CFU/尾及9.6×106CFU/只和4.2×106CFU/只,试验感染鱼出现眼球突出、浑浊及失去方向性等与自然发病相似症状,确定该两株菌为致病菌。两分离株为革兰氏染色阳性,呈链状球菌,β溶血,10℃生长,45℃不生长;接触酶阴性,水解七叶灵、精氨酸;VP试验、脲酶和马脲酸试验阴性,发酵葡萄糖、水杨苷、蔗糖和淀粉,不发醇阿拉伯糖、菊糖、乳糖、蜜二糖、棉子糖和山梨醇。分离株对氨卞青霉素、万古霉素和先锋Ⅴ等高度敏感,对庆大霉素、复方新诺明、林可霉素、氟哌酸等不敏感。应用16SrRNA基因进行的菌株PCR鉴定,确定该两株菌为海豚链球菌。       

A Halophilic Vibrio Isolated from a Case of Chronic Cholecystitis

We found a halophilic vibrio in B bile from a 75-year-old female patient with chronic cholecystitis, and examined its biochemical characteristics. The organisms are gram-negative short rods or comma shaped, with some ring forms. They have a single polar flagellum, but no capsule. The strains can grow in peptone water with 1.0 to 4.0% NaCl, but not with no NaCl or 6.0% NaCl. The characteristics of the organisms are positive dextrose fermentation, catalase, oxidase, and ornithine decarboxylase, and negative lysine decarboxylase, arginine dihydrolase, and absence of gas from glucose. They are sensitive to 2,4-diamine-6,7-diisopropyl pteridine (0/129). These characteristics indicate that the isolated strain should be a halophilic vibrio. However, no growth on Salmonella-Shigella (SS), SS with added sucrose and bromcresol purple (SS-SB), MacConkey's or thiosulfate citrate bile salts (TCBS) agar plates was demonstrated. Nitrate reduction, Simmons' citrate agar, indole, o-nitrophenol-β-d-galactopyranoside (ONPG), motility and esculin hydrolysis were positive. Urease, gelatinase, Voges-Proskauer, phenylalanine deaminase and malonate reactions were negative. Acid was produced from amygdalin, arabinose, cellobiose, fructose, galactose, glucose, lactose, maltose, mannitol, salicin, starch, sucrose, trehalose, and xylose, but not from adonitol, dulcitol, inositol, mannose, melibiose, raffinose, rhamnose and sorbitol. From these characteristics the isolate is considered to be not identical with V. parahaemolyticus, V. cholerae, V. vulnificus or other vibrios. It can be presumed that this isolate represents another species of halophilic vibrio.     

Characterization of Pseudomonas fuscovaginae and Differentiation from Other Fluorescent Pseudomonads Occurring on Rice in Burundi

In various pathogenicity, serological, physiological and biochemical tests, performed in Belgium and Japan, the Pseudomonas fuscovaginae strains associated above 1,350 m elevation in Burundi with sheath brown rot of rice, rusty seed and black, stripes on seedlings, were found to be similar to reference strains of this pathogen from Japan. The same bacteria was detected on rice seeds imported from Asia to Burundi. Beside the serological characteristics, P. fuscovaginae can be differentiated from other oxidase and arginine dihydrolase positive non-pathogenic fluorescent pseudomonads, also isolated from lesions on rice seedlings, by the simultaneous occurrence of no production of 2-ketogluconate, acid production from trehalose, but not from inositol. Occasionally, other symptoms inducing, oxidase positive, fluorescent pseudomonads, different from any described species, were isolated from rice seedlings and sheath rot in Burundi.     

Vibrio aestivus sp. nov. and Vibrio quintilis sp. nov., related to Marisflavi and Gazogenes clades,respectively

Two new Vibrio species, Vibrio aestivus and Vibrio quintilis, are described after a polyphasic characterization of strains M22^T, M61 and M62^T, isolated from seawater collected off a beach on the East coast of Spain (Valencia). All three strains are Gram negative, mesophilic, slightly halophilic, fermentative rods. V. aestivus (M22^T = CECT 7558^T = CAIM 1861^T = KCTC 23860^T and M61 = CECT 7559 = CAIM 1862 = KCTC 23861) is oxidase positive, reduces nitrates to nitrites, is negative for Voges Proskauer, arginine dihydrolase and indole and non hydrolytic on most substrates tested. The 16S rRNA gene sequences of M22^T and M61 are most similar to Vibrio marisflavi (97.1–97.2%) but phylogenetic analysis using NJ, MP and ML methods display Vibrio stylophorae (96.2% similarity) as sibling species. The three species form a deep clade in the genus Vibrio. Average Nucleotide Identity (ANI) values, determined as a measure of overall genomic resemblance, confirmed that strains M22^T and M61 are members of the same species, different to V. marisflavi CECT 7928^T.V. quintilis (M62^T = CECT 7734^T = CAIM 1863^T = KCTC 23833^T) is aerogenic, arginine dihydrolase and Voges Proskauer positive, oxidase negative and unable to reduce nitrate, traits shared by most species in the Gazogenes clade. It is unpigmented and does not grow on TCBS Agar. 16S rRNA gene similarities to its nearest species, Vibrio aerogenes and Vibrio mangrovi, are 97.6% and 96.0% respectively. Strain M62^T and V. aerogenes CECT 7868^T display ANI values well below the 95% boundary for genomic species.     

The widespread occurrence of rutin glycosidase in fluorescent phytopathogenic pseudomonads

Two hundred and twelve strains representing 11 species of fluorescent pseudomonads along with an additional 198 strains isolated from the leaf and root surfaces of plants were tested for rutin glycosidase, on a mineral base medium supplemented with rutin and glucose. AH of the arginine dihydrolase negative phytopathogens (182 strains) with the exception of 12 of 12 strains of Pseudomonas syringae pv. glycina and one of 28 strains of P. syringae pv. phaseoticola showed rutin glycosidase activity whereas few of the arginine dihydrolase positive strains (22 of 228) were positive.     

A rapid method for differentiation of dairy lactic acid bacteria by enzyme systems

Summary A rapid and simple technique utilizing the APIZYM enzymatic patterns complemented with arginine dihydrolase and citratase was developed for species differentiation of 40 lactic acid bacteria relevant to the dairy industry.Streptococcus species in general produced no -galactosidase, except forStreptococcus thermophilus. Lactobacillus species showed strong aminopeptidases and galactosidases but contained no arginine dihydrolase and citratase. Among the group N-streptococci,Streptococcus diacetylactis produced citratase, whereasStreptococcus cremoris differed by the production of butyrate esterase.Streptococcus faecalis was readily distinguishable fromStreptococcus lactis by butyrate esterase activity that was the basis of the differential agar developed. Heterofermentative lactobacilli differed from homofermentative lactobacilli in possessing arginine dihydrolase and citratase but by not producing leucine-aminopeptidase.     

Erwinia and yellow-pigmentedEnterobacter isolates from human sources

Sixty-five strains of gram-negative, yellow-pigmented bacilli, including four chromogenicEnterobacter strains and 55 anaerogenic and six aerogenicErwinia strains, were isolated from human sources. The genusErwinia contained two groups; an anaerogenic group which produced aggregates of bacteria (symplasmata) in the syneresis water of slant cultures and biconvex bodies in colonies on agar medium, and an aerogenic group which lacked these characteristics.Erwinia was differentiated fromEnterobacter, since the latter possessed dihydrolase and decarboxylase activity and demonstrated resistance to cephalothin.     

Subcellular Localization of the Enzymes of the Arginine Dihydrolase Pathway in Trichomonas vaginalis and Tritrichomonas foetus

The enzymes of the arginine dihydrolase pathway were demonstrated in Tritrichomonas foetus and their subcellular localization determined for both T. foetus and Trichomonas vaginalis. Ornithine carbamyltransferase (anabolic and catabolic activities), ornithine decarboxylase and carbamate kinase activity were localized predominately (56–80%) in the non sedimentable fraction of both species. A large proportion (35–40%) of the arginine deiminase was, however, recovered in the large granular fraction, and this distribution was unchanged by increasing the ionic strength of the buffer. Upon density gradient centrifugation the particles containing arginine deiminase activity had an isopycnic density of 1.09 g/ml in percoll, and separated from hydrogenosomes (1.18 g/ml) and lysosomes (1.12 g/ml). Arginine deiminase was also the only enzyme of the dihydrolase pathway which demonstrated latency upon treatment of the 1.09 g/ml fraction with non-ionic detergents. The results demonstrate the presence of the arginine dihydrolase pathway in T. foetus and indicate that at least a portion of the arginine deiminase in trichomonads is membrane associated.     

Synergistic degradation of benzylpenicillin by aPseudomonas strain and aFusarium strain

APseudomonas sp. and aFusarium sp. were isolated from pasture soil. The organisms grew syntrophically on a mineral medium with benzylpenicillin as carbon and nitrogen source. During synergistic degradation of the antibiotic, benzylpenicilloic and benzylpenilloic acid were intermediates. Activities of both organisms were necessary for further decomposition, during which the phenylacetate side chain was degraded. 6-Aminopenicillanic acid did not occur. During syntrophic growth, a red pigment appeared in fungal hyphae growing through bacterial colonies.     

Arginine metabolism inLactobacillus leichmannii

Lactobacillus leichmannii ATCC 4797 metabolizes arginine via the arginine dihydrolase pathway producing ornithine, ammonia, carbon dioxide, and ATP. The specific activities of arginine deiminase and ornithine transcarbamylase were two-or threefold lower (stationary growth phase) in galactose-grown cells. The addition of arginine increased the specific activities of these two enzymes with all growth sugars. When glucose was virtually exhausted from the medium, maximum activities of both enzymes were achieved. The formation of two first enzymes of the arginine dihydrolase pathway inL. leichmannii ATCC 4797 seems to be under the control of two processes: induction by arginine and repression of the induced synthesis by glucose.Dedicated to Dr. Luis F. Leloir on the occasion of his 80th birthday, 6 September 1986.     

西伯利亚鲟(Acipenser baerii)致病性维氏气单胞菌的分离鉴定

摘要：【目的】本研究旨在寻找引起养殖西伯利亚鲟鱼(Acipenser baerii)病害的致病因子。【方法】从北京地区自然患病的西伯利亚鲟鱼体内分离到致病菌株X-1-06909,采用生理生化鉴定结合16S rRNA基因序列的系统发育学分析确定该菌株的系统发育地位。同时采用琼脂扩散法对抗菌类药物的敏感性进行测定。【结果】菌株X-1-06909与Aeromonas veronii ATCC 35624T的16S rRNA基因序列相似性达99.6％;结合形态特征与生理生化测定结果,革兰氏阴性杆菌,具极生单鞭毛     

Modification of sorbitol MacConkey medium containing cefixime and tellurite for isolation of Escherichia coli O157:H7 from radish sprouts

A modified version of sorbitol MacConkey medium containing cefixime and tellurite (CT-SMAC medium) was produced by adding salicin and 4-methylumbelliferyl-beta-D-galactopyranoside to CT-SMAC medium; this medium was designated CT-SSMAC medium and was used to isolate Escherichia coli O157:H7 from radish sprouts. Of 101 non-E. coli bacteria isolated from radish sprouts that produced colorless colonies similar to colonies of E. coli O157:H7 grown on CT-SMAC medium, 92 (91%) formed colonies that were red to pink or were beta-galactosidase negative and colorless on CT-SSMAC medium. On the other hand, colonies of E. coli O157:H7 strains were colorless and beta-galactosidase positive on CT-SSMAC medium. Our results suggest that CT-SSMAC medium is more selective than CT-SMAC medium for isolating E. coli O157:H7.     

Activities of Arginine Dihydrolase and Phosphatase in Streptococcus faecalis and Streptococcus faecium

Strains of Streptococcus faecalis and S. faecium are known to produce ammonia from arginine, but only S. faecalis couples the adenosine triphosphate (ATP) produced through the arginine dihydrolase pathway to growth processes. The specific activities of the arginine dihydrolase enzymes were found to be much lower in S. faecium (0.01 to 0.10) than in S. faecalis (0.24 to 1.60). Phosphatase activities in both strains were similar (up to 0.11), but equaled or exceeded the activities of the arginine dihydrolase enzymes in S. faecium. The failure of S. faecium to show increased growth in arginine media is explained on the basis of low activities of the arginine dihydrolase enzymes coupled with sufficient phosphatase activity to negate any benefit from ATP formed.     

Improved 18-Hour Methyl Red Test

Standard methods for the methyl red (MR) test are not practical for routine use in clinical laboratories because of the necessarily prolonged incubation period. When read after overnight incubation, the usual MR test is often equivocal or falsely positive. The present study demonstrates the importance of standardizing the total volume of broth, the size of the vessel in which the cultures are incubated, and the density of the inoculum. In very small volumes of broth, cultures are better exposed to atmospheric oxygen, and thus MR-negative organisms tend to revert the initial acidic pH much more quickly than in deeper, large-volume broth cultures. In the proposed technique, a single colony was inoculated into a 0.5-ml amount of MR-Voges Proskauer (VP) broth (13- by 100-mm tube), and, after 18 to 24 hr at 37 C, one drop of MR was added. With this technique, the broth cultures produced either a definite red (positive) or yellow (negative) color, whereas various shades of orange were frequently observed when larger volumes of broth were tested after only 1 to 2 days of incubation. With 6.0-ml broth cultures, 18-hr MR tests were totally unreliable, but 18-hr tests in 0.5 ml of broth were comparable to the standard MR test performed after 5 days of incubation and superior to those performed after 48 hr in 6.0-ml broth cultures. With the proposed technique, the MR test can be incorporated readily into the routine scheme for identification of Enterobacteriaceae.     

Occurrence of <Emphasis Type="Italic">Xanthomonas axonopodis</Emphasis> pv. <Emphasis Type="Italic">phaseoli</Emphasis>, the causal agent of common bacterial blight disease,on seeds of common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.) in upper Egypt

Common bean seed lots collected from different seed dealers and Malawii agriculture station were screened for the presence of Xanthomonas axonopodis pv. phaseoli. In the laboratory the pathogen was isolated following the routine laboratory assay method, i.e. direct plating method using yeast extract-dextrose-calcium carbonate agar medium (YDC). Yellow, convex, mucoid colonies of Xanthomonas were consistently isolated on YDC from seed samples. The presumptive pathogen was confirmed by isolation on semiselective medium, such as mTBM and MD5A. Further, the pathogen was confirmed by biochemical, physiological and, finally, the pathogenicity tests. Five samples out of seven were positive for Xanthomonas. The isolates were found to cause common blight of 3-week-old common bean plants by 7 d after inoculation. Bacteria with the same characteristics as those inoculated were re-isolated from the infected plants.     

Presence of the arginine dihydrolase pathway in Mycoplasma

Barile, Michael F. (Division of Biologics Standards, National Institutes of Health, Bethesda, Md.), Robert T. Schimke, and Donald B. Riggs. Presence of the arginine dihydrolase pathway in Mycoplasma. J. Bacteriol. 91:189-192. 1966.-The presence of the arginine dihydrolase pathway was examined in 61 Mycoplasma strains representing at least 18 Mycoplasma species isolated from nine different sources: human, bovine, avian, murine, swine, goat, canine, sewage, and tissue cell culture origin. Some species were represented by only one or two strains. Different strains of the same species gave the same results. Ten species (56%) were positive. Many nonpathogenic Mycoplasma species (M. hominis, type 1 and 2, M. fermentans, M. salivarium, and M. gallinarum) were positive, whereas most pathogenic species (M. pneumoniae, M. gallisepticum, M. neurolyticum, and M. hyorhinis) were negative. The presence of arginine dihydrolase activity among Mycoplasma species may prove to be useful for purposes of identification and classification.     

<Emphasis Type="Italic">Pseudomonas extremaustralis</Emphasis> sp. nov., a Poly(3-hydroxybutyrate) Producer Isolated from an Antarctic Environment

A Gram-negative, mobile, rod-shaped, non-spore-forming bacterium (strain 14-3^T) was isolated from a temporary pond in Antarctica. On the basis of 16S rRNA gene sequence similarity, strain 14-3^T was shown to belong to the genus Pseudomonas sensu stricto. Physiological and biochemical tests supported the phylogenetic affiliation. Strain 14-3^T is closely related to Pseudomonas veronii DSM 11331^T, sharing 99.7% sequence similarity. DNA–DNA hybridization experiments between the two strains showed only moderate reassociation similarity (35.1%). Tests for arginine dihydrolase and nitrate reduction were positive, while those for denitrification, indol production, glucose acidification, urease, ß-galactosidase, esculin, caseine and gelatin hydrolysis were negative. Growth of this bacterium occurred in a range from 4 to 37°C but not at 42°C. It accumulated poly(3-hydroxybutyrate) when grown on sodium octanoate medium. Strain 14-3^T therefore represents the type strain of a new species, for which the name Pseudomonas extremaustralis sp. nov. is proposed. The type strain 14-3^T has been deposited as DSM 17835^T and as CIP 109839^T.     

Cholesterol-reducing bacterium from human feces.

An anaerobic, gram-positive diplobacillus that reduces cholesterol to coprostanol was isolated from human feces and rat cecal contents. The isolates closely resemble a cholesterol-reducing organism isolated by Eyssen et al. (H. Eyssen et al., Eur. J. Biochem. 36:412-421, 1973) from a rat's cecum. These organisms would not form colonies and were isolated and cultivated in an anaerobic medium containing homogenized pork brains (naturally high in cholesterol). These organisms require free or esterified cholesterol for growth. They were isolated by serially diluting feces or cecal contents and inoculating brain medium. Colony-forming organisms, which did not reduce cholesterol, were eliminated by addition of inhibitory agents to the brain medium cultures. This serial dilution procedure was performed until a pure culture of a cholesterol-reducing organism was obtained.     

Cholesterol-reducing bacterium from human feces.

An anaerobic, gram-positive diplobacillus that reduces cholesterol to coprostanol was isolated from human feces and rat cecal contents. The isolates closely resemble a cholesterol-reducing organism isolated by Eyssen et al. (H. Eyssen et al., Eur. J. Biochem. 36:412-421, 1973) from a rat's cecum. These organisms would not form colonies and were isolated and cultivated in an anaerobic medium containing homogenized pork brains (naturally high in cholesterol). These organisms require free or esterified cholesterol for growth. They were isolated by serially diluting feces or cecal contents and inoculating brain medium. Colony-forming organisms, which did not reduce cholesterol, were eliminated by addition of inhibitory agents to the brain medium cultures. This serial dilution procedure was performed until a pure culture of a cholesterol-reducing organism was obtained.