Acid and alkali as aids in differentiation of pathogenic Nocardia and for their isolation from clinical specimens

The tolerance of Nocardia brasiliensis, Nocardia asteroides and Nocardia caviae to different concentrations of acids and alkalis was studied in this work. The purpose was to determine their use: (a) in the differentiation of species; (b) for decontamination of clinical materials during the isolation of pathogenic Nocardia. The results showed (1) that both 2 M NH₃ and 0.05 M H₂SO₄ solutions are useful for differentiating species. They can also be used as a complementary method for identification. (2) That both 1 M NH₃ and 0.05 M H₂SO₄ solutions were useful for decontamination of clinical materials during the isolation of pathogenic Nocardia.     

Natural occurrence of Nocardia in soil of Tucumán: Physiological characteristics

This is the first study initiated in Argentina to establish the presence of species of Nocardia from soil samples. These samples were gathered in different areas of Tucumán.Thirty three pathogenic strains of Nocardia were isolated by the paraffin bait method. Out of them, 28 were N. brasiliensis, 3 N. asteroides and 2 N. caviae. N. brasiliensis was widely distributed in the soil of the areas tested.It is proved that N. caviae, so rarely found in other regions of the world, occurs in Tucumán.A detailed study of the morphological and physiological characteristics for identification is discussed.     

Phospholipid and acid composition of Nocardia and nocardoid bacteria as criteria of classification

Phospholipid and acid composition of 5 strains of ‘true’ Nocardia and 4 strains of nocardoid bacteria have been studied. A great homogeneity was found in all the Nocardia species: phospholipids consist of cardiolipin, phosphatidyl ethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. Streptomyces (Nocardia) mediterranei did not contain phosphatidylinositol and Oerskovia (Nocardia) turbata had no phosphatidyl ethanolamine. The fatty acid composition of these phospholipids was determined and was found different in Nocardia and nocardoid species. Nocardia were rich in straight chain fatty acids and tuberculostearic acid while the phospholipids of nocardoid bacteria contained greater amounts of branched fatty acids. The fatty acids from acetone soluble lipids consisted of hydroxy and non-hydroxy compounds. Hydroxy acids were found in Nocardia which contained nocardic acids: high MW β-hydroxy α-branched acids and in S. mediterranei which contained β-hydroxy acids with 15–17 carbon atoms. Non-hydroxy acids were essentially palmitic and tuberculostearic acids in Nocardia species while S. mediterranei and O. turbata contained great amounts of iso acids from C₁₄ to C₁₇. Phospholipid and acid composition are discussed as criteria of taxonomic classification of Nocardia and related Actinomycetes.     

Use of paraffin bait technique in the isolation of Nocardia asteroides from sputum

Summary The use of paraffin bait technique in the isolation ofNocardia asteroides has been tested in 241 samples of sputa obtained from 235 cases of respiratory diseases.N. asteroides was recovered on 6 occasions from sputum of a patient using the paraffin bait technique. On the other hand cultures of sputa from the same patient on routine agar media such as Sabouraud's agar and Lowenstein Jensen medium yielded only one isolated of the pathogen.This work forms a part of the Ph. D. thesis of P.V.K. submitted to the University of Delhi.     

Identification of Nocardia and non-tuberculous Mycobacterium species by MALDI-TOF MS using the VITEK MS coupled to IVD and RUO databases

Identification of Nocardia and Mycobacterium species by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) is still a challenging task that requires both suitable protein extraction procedures and extensive databases. This study aimed to evaluate the VITEK MS Plus system coupled with updated RUO (v4.17) and IVD (v3.2) databases for the identification of Nocardia spp. and Mycobacterium spp. clinical isolates. Sample preparation was carried out using the VITEK MS Mycobacterium/Nocardia kit for protein extraction. From 90 Nocardia spp. isolates analysed, 86 (95.6%) were correctly identified at species or complex level using IVD and 78 (86.7%) using RUO. Only two strains were misidentified as other species pertaining to the same complex. Among the 106 non-tuberculous Mycobacterium clinical isolates tested from a liquid culture medium, VITEK MS identified correctly at species or complex level 96 (90.6%) isolates in the IVD mode and 89 (84.0%) isolates in the RUO mode. No misidentifications were detected. Although the IVD mode was unable to differentiate members of the M. fortuitum complex, the RUO mode correctly discriminated M. peregrinum and M. septicum. The robustness and accuracy showed by this system allow its implementation for routine identification of these microorganisms in clinical laboratories.     

Genome based analysis of type-I polyketide synthase and nonribosomal peptide synthetase gene clusters in seven strains of five representative Nocardia species

Background

Actinobacteria of the genus Nocardia usually live in soil or water and play saprophytic roles, but they also opportunistically infect the respiratory system, skin, and other organs of humans and animals. Primarily because of the clinical importance of the strains, some Nocardia genomes have been sequenced, and genome sequences have accumulated. Genome sizes of Nocardia strains are similar to those of Streptomyces strains, the producers of most antibiotics. In the present work, we compared secondary metabolite biosynthesis gene clusters of type-I polyketide synthase (PKS-I) and nonribosomal peptide synthetase (NRPS) among genomes of representative Nocardia species/strains based on domain organization and amino acid sequence homology.

Results

Draft genome sequences of Nocardia asteroides NBRC 15531^T, Nocardia otitidiscaviarum IFM 11049, Nocardia brasiliensis NBRC 14402^T, and N. brasiliensis IFM 10847 were read and compared with published complete genome sequences of Nocardia farcinica IFM 10152, Nocardia cyriacigeorgica GUH-2, and N. brasiliensis HUJEG-1. Genome sizes are as follows: N. farcinica, 6.0 Mb; N. cyriacigeorgica, 6.2 Mb; N. asteroides, 7.0 Mb; N. otitidiscaviarum, 7.8 Mb; and N. brasiliensis, 8.9 - 9.4 Mb. Predicted numbers of PKS-I, NRPS, and PKS-I/NRPS hybrid clusters ranged between 4–11, 7–13, and 1–6, respectively, depending on strains, and tended to increase with increasing genome size. Domain and module structures of representative or unique clusters are discussed in the text.

Conclusion

We conclude the following: 1) genomes of Nocardia strains carry as many PKS-I and NRPS gene clusters as those of Streptomyces strains, 2) the number of PKS-I and NRPS gene clusters in Nocardia strains varies substantially depending on species, and N. brasiliensis strains carry the largest numbers of clusters among the species studied, 3) the seven Nocardia strains studied in the present work have seven common PKS-I and/or NRPS clusters, some of whose products are yet to be studied, and 4) different N. brasiliensis strains have some different gene clusters of PKS-I/NRPS, although the rest of the clusters are common within the N. brasiliensis strains. Genome sequencing suggested that Nocardia strains are highly promising resources in the search of novel secondary metabolites.

Electronic supplementary material

The online version of this article (doi: 10.1186/1471-2164-15-323) contains supplementary material, which is available to authorized users.     

Physiological and pathogenic characteristics of Nocardia brasiliensis isolated from human mycetomas

A previous analysis of the physiological properties of Nocardia brasiliensis strains isolated from soil of Tucumán proves that non-pathogenic strains have a different behaviour pattern from the pathogenic strains.In the present paper, 16 Nocardia brasiliensis strains isolated from human mycetomas were studied in the same way. The object is to determine if any of the Nocardia brasiliensis present in soil can produce human mycetomas.The macro and micromorphological, biochemical (17 tests), physiological (4 tests) and pathological characteristics were determined for each of the strains. Experimental pathogenicity was determined using albino Swiss mice by inoculation into the footpads.The strains of Nocardia brasiliensis that cause human mycetomas have the same physiological pattern and experimental pathogenicity as the virulent strains present in soil.     

Studio di una nuova specie di Streptomyces: Streptomyces nobilis sp. nov. e esame di ceppi affini appartenenti ai generi streptomyces,Streptoverticillium e Nocardia

Riassunto Alcuni ceppi di attinomiceti provenienti da diverso isolamento sono stati esaminati in comparazione con altri riferiti a specie o generi noti, onde procedere alla loro identificazione. I criteri diagnostici sono gli stessi di cui a precedenti nostre pubblicazione.I ceppi in studio sono riferibili a tre generi come da seguito: GenereStreptomyces. Sono riferiti aStreptomyces i ceppi 631 e 725, isolati da suolo, e sono descritti come specie nuovaStreptomyces nobilis sp. nov. Il ceppo 631 costituisce l'olotipo e il ceppo 725 una variante naturale.GenereStreptoverticillium. Viene riferito il ceppo in studio 174. Si è rinunziato ad una identificazione specifica limitando si all'inserimento nella serie Rubrireticuli della quale viene aggiornata la chiave delle specie.GenereNocardia. I ceppi 608 e 669 sono riferiti aNocardia asteroides e il ceppo 535 aProactinomyces (=Nocardia)ruber.

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Summary Strains of actinomycetes, isolated from different sources, have been examined comparatively together with cultures of known species and genera, in order to proceed to their identification. The diagnostic criteria followed are those described in previous papers.Our isolates have been referred to the following genera:GenusStreptomyces: to this genus belong our strains 631 and 725, isolated from soil and described as new species,Streptomyces nobilis sp. nov. Strain 631 is considered as the holotype and strain 725 as a natural variant.GenusStreptoverticillium: strain 174 belongs to this genus. No specific identification has been given, but the strain has been enclosed in the Rubrireticuli Series. A new key of the Series is provided.GenusNocardia: strains 608 and 669 are referred toNocardia asteroides, strain 535 toProactinomyces (=Nocardia)ruber.

     

Nocardia rhamnosiphila sp. nov., isolated from soil

In this study two actinomycete strains were isolated in Cape Town (South Africa), one from a compost heap (strain 202GMO^T) and the other from within the fynbos-rich area surrounded by the horseracing track at Kenilworth Racecourse (strain C2). Based on 16S rRNA gene sequence BLAST analysis, the strains were identified as members of the genus Nocardia. Phylogenetic analysis showed that the strains clustered together and are most closely related to Nocardia flavorosea NRRL B-16176^T, Nocardia testacea JCM 12235^T, Nocardia sienata IFM 10088^T and Nocardia carnea DSM 43397^T. This association was also supported by gyrB based phylogenetic analysis. The results of DNA–DNA hybridization and physiological tests allowed genotypic and phenotypic differentiation of both strains 202GMO^T and C2 from related species. However, their high DNA relatedness showed that they belong to the same species. Strain 202GMO^T was selected as the type strain to represent this novel species, for which the name Nocardia rhamnosiphila is proposed (=DSM 45147^T = NRRL B-24637^T).     

Nocardia bhagyanesis sp. nov., a novel actinomycete isolated from the rhizosphere of Callistemon citrinus (Curtis), India

A novel actinomycete strain, designated VRC07^T, was isolated from a Callistemon citrinus rhizosphere sample collected from Hyderabad, India. Its taxonomic status was determined by using polyphasic approach. It is a Gram-positive, aerobic, non-motile, weakly acid-fast strain. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain VRC07^T is a member of the genus Nocardia. The highest levels of 16S rRNA gene sequence similarity was found between the strains Nocardia niwae W9241^T (99.6 %), Nocardia amikacinitolerans W9988^T (99.3 %) and Nocardia arthritidis IFM 10035^T (98.9 %); similarity to other type strains of the genus Nocardia was below 98.7 %. The organism had chemical and morphological features consistent with its classification in the genus Nocardia such as meso-diaminopimelic acid as the diagnostic diamino acid in the cell wall peptidoglycan. Arabinose and galactose as the diagnostic sugars. Diagnostic polar lipids were phosphatidylinositol, diphosphatidylglycerol, and phosphatidylglycerol. The predominant menaquinone was MK-8(H_{4, ω-cycl}). The major fatty acids were C_16:0, C_18:0, C_{18:1 w9c}, C_{18:0 10-methyl TBSA} and sum in feature 3 (16:1 w7c/16:1 w6c). The G+C content of the genomic DNA was 68.5 mol%. The DNA–DNA relatedness data, together with phenotypic differences clearly distinguished the isolate from its closest relatives. On the basis of these phenotypic and genotypic data, the isolate represents a novel species, for which the name Nocardia bhagyanesis sp. nov., is proposed. The type strain is VRC07^T (=KCTC 29209^T = MTCC 11725^T = ATCC BAA-2548).     

Effect of Rifampicin and Uracil Depletion on Bacteriophage ϕX174 DNA Synthesis in an E. coli dnaHts Mutant

The fluorescent antibody technique was used to trace an inoculated Nocardia erythropolis strain which was capable of rapidly degrading phthalate esters in soil column and activated sludge systems. The reaction of antibody to Nocardia erythropolis S-1 was highly strain specific, i. e., only one of twelve other strain of N. erythropolis was stained with this fluorescent antibody. All other species of Nocardia and other genera of bacteria and a strain of Candida were not stained. Using this technique it was demonstrated that N. erythropolis S-1 inoculated into activated sludge and soil column systems was successfully distinguished from many other microorganisms in mixed culture systems, and the distribution of this strain was appreciated.     

Observations on paraffin baiting as a laboratory diagnostic procedure in nocardiosis

The efficacy of paraffin bait technique in the isolation ofNocardia asteroides from clinical specimens has been investigated. In a comparative study 1091 clinical specimens, mostly sputa and bronchial aspirates collected from 639 patients of bronchopulmonary diseases and 11 of meningitis, were examined by paraffin baiting and the conventional technique. Thirty-six clinical specimens originating from 12 of the patients yieldedN. asteroides by the paraffin bait technique but only 4 by the conventional technique. Approximately 95 % of 125 sputum samples inoculated withN. asteroides yielded the pathogen by paraffin baiting as against 49 % by the conventional technique. Paraffin baiting was more productive than the conventional technique in the isolation ofN. asteroides from mixed suspensions with a number of fungi and bacteria. It is concluded that paraffin baiting can be profitably adopted as a suitable technique for the isolation ofN. asteroides from clinical specimens, such as, sputum, gastric lavage, etc., which are often contaminated. The technique has no particular advantage with non-contaminated specimens.This work forms a part of the Ph. D. thesis of S.K.M. submitted in 1971 to the University of Delhi, and was presented at the Vth meeting of ISHAM held in Paris from 5th–10th July, 1971.     

Photochromogenesis

The effect of light on the pigmentation of various strains of Nocardia, Corynebacterium, Arthrobacter, Brevibacterium, and Flavobacterium was investigated. It was revealed that thirty out of fifty-seven strains of Nocardia, two out of fifteen strains of Corynebacterium, two strains of Arthrobacter, six out of thirteen strains of Brevibacterium and two out of fourteen strains of Flavobacterium were photochromogenic; i. e., these strains produced yellow or orange pigments when grown under illumination but entirely unpigmented in total darkness. From these results, it may be concluded that photochromogenicity is not a particular phenomenon limited to specific species, but a common, widely distributed phenomenon in nonphotosynthetic bacteria.     

Nocardia gamkensis sp. nov.

A novel actinomycete, strain CZH20^T, was isolated from a soil sample taken from the banks of the Gamka River in the Swartberg Nature Reserve, Western Cape Province, South Africa. Strain CZH20^T was identified as a member of the genus Nocardia by a polyphasic approach. Strain CZH20^T could be differentiated from other members of the genus Nocardia on the basis of physiology and 16S-rRNA gene sequence analysis. It exhibited weak antibiosis against Mycobacterium aurum A+. Organic solvent extracts of the culture filtrate and mycelial mass of CZH20^T exhibited moderate antibiotic activity against Mycobacterium smegmatis LR222 and Mycobacterium tuberculosis H37Rv. The name Nocardia gamkensis is proposed, with the type strain CZH20^T (=DSM 44956^T =NRRL B-24450^T).The GenBank accession number for the 16S-rRNA-gene sequence of Nocardia gamkensis CZH20^T is DQ235272.     

Nocardia hydrocarbonoxydans n. spec., ein oligocarbophiler Actinomycet

Zusammenfassung Eine neue Actinomycetenart (Stamm 70), aufgetreten als Luftverunreinigung einer Kieselgelplatte, wird beschrieben.Auf Grund der Fähigkeit, mit aliphatischen Kohlenwasserstoffen aus der Gasphase zu leben, soll die neue Art Nocardia hydrocarbonoxydans genannt werden. Sie ist von anderen, bereits bekannten oligocarbophilen Actinomycetenarten durch ihre Kolonieform, durch die Fähigkeit Hexan, Heptan und Octan zu oxydieren, und durch ihr Unvermögen, auf Glycerin-Kartoffelkeilen, Kartoffelkeilen oder Blut-Agar zu wachsen, wohl unterschieden. Ihre Stellung im System und die Verwandtschaft mit anderen Arten der Gattung Nocardia wird diskutiert.

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Summary A new actinomycete species, and aerial contamination on a silica gel plate, is described. Because of its ability to utilize gaseous aliphatic hydrocarbons (C₆–C₁₄) for growth, Nocardia hydrocarbonoxydans is proposed as the name of the new species. It differs from other oligocarbophilic actinomycetes in its colony form, its ability to oxidize hexane, heptane and octane, and its inability of growing on glycerol potatoe slants, potatoe slants or blood agar. Taxonomical relationships to species of the genus Nocardia are discussed.

     

An ultrastructural comparison of the cell envelopes of selected strains of Nocardia asteroides and Nocardia brasiliensis

Growth curves were determined for three strains each ofNocardia asteroides andNocardia brasiliensis. Two strains ofN. brasiliensis and one strain ofN. asteroides had longer lag periods of growth than the remaining three strains. All strains had generation times of approximately 5.5 hours.The ultrastructure of the cell envelope of eachNocardia strain in early stationary phase growth was also examined. All the strains had typical trilaminar cell walls and cell membranes. The thickness of the cell wall layers, especially the inner peptidoglycan layer, varied from strain to strain. The inner layer of two strains ofN. brasiliensis and one strain ofN. asteroides was 12 nm or more in thickness, while that of the remaining three strains was 7 nm thick. These observed differences in growth patterns and/or thickness of the cell wall layers could be correlated to the varying degress of virulence as well as the divergent pathologies exhibited by these organisms.     

诺卡氏菌烈性噬菌体vB_Ncarnea_KYD1的分离纯化与基因组分析

【背景】诺卡氏菌是一种广泛分布的好氧放线菌,可在人体内引起局部或播散性感染,尤其是在免疫功能低下的个体中。诺卡氏菌感染在临床上较难鉴定,而且不断有新型诺卡氏菌种被发现。不同类型、不同地域的诺卡氏菌具有流行差异和抗生素敏感性差异,阻碍了适当治疗方式的选择。利用病灶处的宿主菌分离得到噬菌体来控制诺卡氏菌感染的这种方法在近年来受到了各界的关注。【目的】尝试从环境中分离出能够用于临床治疗的针对诺卡氏菌的烈性噬菌体,并研究其基因组学特征。【方法】利用双层平板法分离得到目标噬菌体,观察其噬菌斑形态,并对噬菌体进行分离纯化,在透射电镜下鉴定其特征。提取噬菌体DNA进行全基因组测序与注释,并与数据库内已知噬菌体基因组进行比较,同时构建系统进化树以进行遗传进化分析。【结果】本文以肉色诺卡氏菌为宿主,从环境样本中分离出一株烈性噬菌体vB_Ncarnea_KYD1,在双层平板上可形成直径<2 mm的透亮均匀的噬菌斑。基因组分析表明,vB_Ncarnea_KYD1DNA为环状,大小为66 621 bp,共发现102个蛋白质编码区(coding sequence,CDS)及一个tRNA-Ser编码序列。透射电镜观察与系统进化树综合分析可以确定,vB_Ncarnea_KYD1为长尾噬菌体科的一个新属。其在进化过程中经历了复杂的基因重组过程。暂未发现毒力因子相关基因与抗性基因,具备实用价值。【结论】从环境水体中分离出一株烈性肉色诺卡氏菌噬菌体vB_Ncarnea_KYD1,通过电镜观察与基因组分析可知,此株噬菌体为长尾噬菌体,基因组中暂未发现不利于临床应用的相关基因,是一株相对安全的烈性诺卡氏菌噬菌体。研究结果丰富了国内噬菌体资源库,并为后续诺卡氏菌感染疾病的治疗提供支持。     

Molecular Identification of Nocardia Isolates from Clinical Samples and an Overview of Human Nocardiosis in Brazil

Background

Nocardia sp. causes a variety of clinical presentations. The incidence of nocardiosis varies geographically according to several factors, such as the prevalence of HIV infections, transplants, neoplastic and rheumatic diseases, as well as climate, socio-economic conditions and laboratory procedures for Nocardia detection and identification. In Brazil the paucity of clinical reports of Nocardia infections suggests that this genus may be underestimated as a cause of human diseases and/or either neglected or misidentified in laboratory specimens. Accurate identification of Nocardia species has become increasingly important for clinical and epidemiological investigations. In this study, seven clinical Nocardia isolates were identified by multilocus sequence analysis (MLSA) and their antimicrobial susceptibility was also determined. Most Nocardia isolates were associated to pulmonary disease.

Methodology/Principal Findings

The majority of Brazilian human isolates in cases reported in literature were identified as Nocardia sp. Molecular characterization was used for species identification of Nocardia nova, Nocardia cyriacigeorgica, Nocardia asiatica and Nocardia exalbida/gamkensis. Data indicated that molecular analysis provided a different Nocardia speciation than the initial biochemical identification for most Brazilian isolates. All Nocardia isolates showed susceptibility to trimethoprim-sulfamethoxazole, the antimicrobial of choice in the treatment nocardiosis. N. nova isolated from different clinical specimens from one patient showed identical antimicrobial susceptibility patterns and two distinct clones.

Conclusions/Significance

Although Brazil is the world''s fifth-largest country in terms of land mass and population, pulmonary, extrapulmonary and systemic forms of nocardiosis were reported in only 6 of the 26 Brazilian states from 1970 to 2013. A least 33.8% of these 46 cases of nocardiosis proved fatal. Interestingly, coinfection by two clones may occur in patients presenting nocardiosis. Nocardia infection may be more common throughout the Brazilian territory and in other developing tropical countries than is currently recognized and MLSA should be used more extensively as an effective method for Nocardia identification.     

不同分离方法对子实体形成和粘细菌分离的影响

【目的】基于模拟原位环境策略、可培养粘细菌的营养策略及细菌互作网络,改良分离培养基,以提高分离粘细菌的多样性。【方法】通过添加土壤浸提液、使用不同种类的诱导菌和改变诱导菌的接种方式设置分离方法,同时以传统的分离方法作对照。【结果】改良的分离方法比对照组诱导出了更多粘细菌子实体种类,采自4个地区的9份样品共分离纯化出40株粘细菌,按形态学和分子生物学,将其归类于原囊菌属(Archangium)、珊瑚菌属(Corallococcus)、软骨霉状菌属(Chondromyces)、粘球菌属(Myxococcus)、侏囊菌属(Nannocystis)、多囊菌属(Polyangium)、匣状球菌属(Pyxidicoccus)。【结论】与传统分离方法相比,添加土壤浸提液,诱导菌点接法能大大提高诱导出的粘细菌子实体种类的数目,革兰氏阳性菌和革兰氏阴性菌作为诱导菌对子实体种类影响较小,但是也发现革兰氏阳性菌特异性诱导出的子实体。虽然本研究通过对分离培养基的改良大大增加了子实体种类,但是纯化出的粘细菌种类远少于观察到的子实体种类,说明除改良分离方法外,还需进一步研究粘细菌的纯化方法,提高分离所得粘细菌的多...     

Study on the role of Nocardia farcinica in enhancing the flow rate of crude oil

Microbial degradation of paraffin wax is an efficient method of removing wax deposition from pipelines and enhancing the flow rate of crude oil. The present study was carried out to isolate a potential paraffin-degrading organism from oil wells of Gujarat. Screening for bacteria-utilizing paraffin wax as the sole source of carbon was carried out using 2,6-dichlorophenol indophenol (DCPIP) dye as redox indicator. The selected organism was identified as Nocardia farcinica by 16S rRNA sequencing. Nocardia farcinica showed 100% degradation of heneicosane, 65.99% degradation of docosane, and 50.59% degradation of tricosane, the major components of paraffin wax, in 8 days, which was observed by gas chromatography. Eicosane (86%) and heneicosane (80%) were utilized more by the selected organism compared with octacosane (61%) and triacontane (58%) (DCPIP dye method). Gas chromatographic analysis revealed that the selected organism degraded 50% of paraffin crude oil in 10 days. To determine the ability of the selected organism to enhance flow rate, parameters such as viscosity (cps), surface tension (d/cm²), pour point (°C), and flow rate (min/2 ml) were determined, and the result showed significant reduction in all the parameters after the addition of Nocardia farcinica. The viscosity and surface tension of crude oil were reduced by 22 and 6.30 points, respectively, after the addition of Nocardia farcinica. Pour point and flow rate were reduced by 2 and 11 points, respectively, when compared with control. The above findings indicate that Nocardia farcinica isolated from crude oil plays a major role in enhancing the flow rate of crude oil.