Brain abscess due to Nocardia

Cerebral Nocardiosis is a rare disease, usually occurring in immunocompromised hosts. We report here two cases of brain abscess due to Nocardia species-one due to usual N. asteroides and other by uncommon N. caviae. N. asteroides affected the brain in a post renal transplant patient, whereas N. caviae caused infection of brain in an apparently healthy individual. To the best of our knowledge, all the previous cases of brain abscess due to Nocardia caviae have been reported in compromised hosts. Agar dilution antimicrobial testing showed relatively higher resistant pattern in N. asteroides. In spite of antimicrobial therapy, both the patients succumbed, one within 4 days and other after an initial improvement for four weeks due to drainage of abscess.The technical assistance of Mr. R.K. Sapra and Mr. Pawan Kumar is greatfully acknowledged.     

Nocardiosis in a tertiary care hospital in North India and review of patients reported from India

Nocardiosis, an uncommon infection of the past, is being increasingly reported in recent years with rise of immunosuppressed patients. In India, very few centers have reported this disease. The present report describes twelve consecutive cases of nocardiosis reported over a period of 26 months (January 2004 to March 2006) from a tertiary care center in north India. The patients were predominantly males (75%) with age range of 8–65 years and mean age of 38.4 years. Eleven patients had known underlying illness including renal transplantation, human immunodeficiency virus infection, and long-term steroid therapy. One patient with cerebral nocardiosis had no detectable predisposing factor. Infection involved central nervous system (3 cases), lungs (5 cases), subcutaneous tissue (1 case), and anterior mediastinum (1 case). Disseminated infection was documented in two patients. Nocardia asteroides complex was incriminated in six patients, N. brasiliensis in five and N. otitidiscaviarum in one patient. All the isolates were sensitive to co-trimoxazole, amikacin, cefotaxime and imipenem. Good therapeutic response was observed in 10 (90.9%) of 11 patients treated with antibiotics including co-trimaxazole, cephalosporins, amikacin, and imipenem alone or in combination. The series of nocardiosis reported from India has also been reviewed.     

First reportedNocardia otitidiscaviarum infection in an AIDS patient in Italy

Nocardiosis is a well-described infection in immunocompromised patients, and has been rarely documented in patients with AIDS.Nocardia asteroides is the most frequently isolated etiologic agent. Rare cases are due toN. brasiliensis andN. otitidiscaviarum. This work describes the first case of nocardiosis in Italy caused byN. otitidiscaviarum in an AIDS patient. A 31 year-old intravenous drug abuser with a diagnosis of full-blown AIDS, presented with high fever and lymphadenitis with a fistula draining copious purulent discharge. Broad-spectrum antibiotic therapy was initiated, but the patient did not show any improvement. Direct examination of the pus revealed numerous gram positive rods. When culturedN. otitidiscavarium was isolated and identified by morphological and biochemical tests.     

Subcutaneous Pseudallescheriasis in a renal transplant recipient

This paper reports a case of a single subcutaneous nodule caused byPseudallescheria boydii in a renal transplant recipient, possibly of nontraumatic origin. The patient was treated surgically and with itraconazole.     

Application of the API YeastIdent system in determining the enzymatic profiles ofNocardia asteroides isolates

Glucose represses production of ammonium in many clinical isolates ofNocardia asteroides growing on bromcresolpurple casein glucose agar. Strains exhibiting this property are designated as group A, while group B represents isolates showing a high degree of proteolytic activity and a resulting rapid increase in pH. Twenty isolates ofN. asteroides were characterized as group A or B. Enzymatic profiles obtained using the API YeastIdent system showed significant enzymatic variation between 12 group B and 8 group A isolates. Proteolytic enzymes which most varied in activity between groups were glycine aminopeptidase, histidine aminopeptidase and leucyl glycine aminopeptidase. As some of theN. asteroides isolates were isolated from asymptomatic patients, it is of interest to consider the possibility of one group being of low virulence while the other is more strongly associated with infection.     

Effect of the Live and Heat-Killed Cells of Staphylococcus aureus on the Resistance of Mice to the Infection with Nocardia asteroides

When living cells of Staphylococcus aureus were introduced into mice by various routes within a few hours before or after infection with Nocardia asteroides, a marked increase in mortality and enhanced severity of lesions were observed. This effect was presumably caused by a heat-labile substance(s) liberated from the cells of S. aureus. No increase in resistance occurred when S. aureus was given 2 to 7 days before the nocardial infection, contrary to the effect of E. coli as reported previously. An increased resistance was noted when heat-killed cells of E. coli were administered two to four days before concurrent infection with N. asteroides and S. aureus.     

Antibodies to phospholipids in experimental nocardiosis

Live Nocardia asteroides injected intraperitoneally into guinea pigs produced antibodies to phospholipids of Nocardia whereas antibodies were not detected in animals injected with heat-killed N. asteroides. The antibody titer was found to be related with the degree of infection; a significant decrease was noted after sulfadiazine treatment suggesting that antibodies were produced in response to an increasing bacterial load that occurred as the infection progressed.This work was partly financed by a grant from the Indian Council of Medical Research.     

Cloning vector system forNocardia spp.

An electro-transformation system has been developed forNocardia asteroides andNocardia corallina by using aNocardia-Escherichia coli shuttle vector. The shuttle vector, named pCY104, was constructed by joining a 2.5-kb crypticN. asteroides plasmid pCY101 with theE. coli plasmid pIJ4625. The resistance genes for kanamycin, chloramphenicol, and thiostrepton on plasmid pCY104 were expressed inN. asteroides andN. corallina. The transformation method was optimized forN. asteroides, and transformation efficiency of 8×10⁴ transformants per g plasmid DNA was achieved routinely.     

Subcutaneous Mycotic Cyst Caused by <Emphasis Type="Italic">Roussoella percutanea</Emphasis> in a UK Renal Transplant Patient

Fungi from more than 100 genera have been implicated in subcutaneous fungal infections, usually following traumatic inoculation of the etiologic agent. With the advent of molecular approaches to fungal identification and taxonomy, novel agents of subcutaneous mycoses are increasingly reported. In this manner, Roussoella percutanea, a novel species in Pleosporales, was described in 2014 from a subcutaneous mass in an immunocompetent male adult. Two additional cases involving renal transplant recipients were recently reported from patients resident in France and Germany, with several further cases discovered after analyses of historical culture collection isolates. Here, we describe a new case of subcutaneous R. percutanea infection, causing a mycotic cyst in a renal transplant patient resident in the UK. Although fungal infection was confirmed histologically, viable fungal isolates could not be recovered in culture from biopsy material and identification of the causative agent relied upon PCR amplification and sequencing of fungal rDNA genes. This is the fourth well-documented case of infection with R. percutanea in renal transplant patients, and the first reported from a patient resident in the UK. The current case illustrates the importance of molecular approaches for the identification of emerging fungal pathogens in culture-negative subcutaneous fungal infections.     

Numerical Analysis of the Taxonomy of Nocardiae and Rhodococci

The genera Nocardia and Rhodococcus were clearly differentiated in the present study. Eleven characteristics were shown to be useful for differentiation between these two genera. Nocardia asteroides sunsu stricto previously defined by Tsukamura was divided into two taxa. One contained the type strain and was considered to retain the name Nocardia asteroides in a new sense. Another was named in the present study as Nocardia nova sp. nov. Tsukamura. The type strain of this species is ATCC 33726. The following seven characters were useful for differentiating N. nova from newly defined N. asteroides: 1) arylsulfatase activity after 14 days; 2) catalase activity (semiquantitative); 3) β-esterase activity; 4) pyrazinamidase activity; 5) utilization of citrate as a sole source of carbon; 6) utilization of 2,3-butylene glycol as a sole carbon source; and 7) resistance to 5-fluorouracil (20 μg/ml). The name Nocardia farcinica for Tsukamura's Kyoto-I group should be rejected. This taxon has been named Nocardia paratuberculosis sp. nov. Tsukamura. The type strain is ATCC 23826. Three new species of the genus Rhodococcus were proposed: Rhodococcus aichiensis sp. nov. Tsukamura (type strain, ATCC 33611); Rhodococcus chubuensis sp. nov. Tsukamura (type strain, ATCC 33609); Rhodococcus obuensis sp. nov. Tsukamura (type strain, ATCC 33610).     

Effect of presensitization of Nocardia asteroides with specific antibody on the viability of the organism

Nocardia asteroides from various growth phases was treated in vitro with normal rabbit sera, immune rabbit sera containing nocardial polyclonal antibodies and a monoclonal antibody. At intervals, samples were grown in broth or on blood agar plates to determine their viability. Log and stationary phase cells were injected intra-peritoneally into female BALB/c mice and their survival rates in the liver and spleen were determined. Presensitization with antibodies reduced the viability of the log phase cells by 48% and that of the late stationary phase by 4%. The antibody-treated log phase organisms were less viable on the blood agar medium and in the spleen and liver than the control organisms. This indicates that pretreatment with antibody has a lethal effect on N. asteroides and affects its survival in vivo.     

Nocardia asteroides culture filtrates cause dopamine depletion and cytotoxicity in PC12 cells

Experimental infection of BALB/c mice with the gram-positive bacterium Nocardia asteroides produces marked loss of nigrostriatal dopamine neurons, resulting in striatal dopamine depletion. To investigate the mechanism(s) responsible for this neuronal loss, we evaluated the influence of N. asteroides cell-free culture filtrates on rat pheochromocytoma PC12 cells, an in vitro model for dopamine neurons. Changes in cell viability and cell numbers were minimal after 24 h, but increased with longer incubation. In contrast, dopamine depletion occurred after 30 min incubation, and was greater with GUH-2 filtrate than with filtrate from the less virulent strain 10905. Incubation with the culture filtrate decreased viability in neuroblastoma and glioma cell lines, indicating that cytotoxic effects were not limited to dopaminergic cells. These findings suggest that the loss of nigrostriatal dopamine neurons and concomitant striatal dopamine depletion in Nocardia-infected mice may be due, at least in part, to the neurotoxicity of nocardial secretory products.     

Isavuconazole and Liposomal Amphotericin B as Successful Combination Therapy of Refractory Invasive Candidiasis in a Liver Transplant Recipient: A Case Report and Literature Review

Invasive fungal infections in liver transplant recipients are associated with elevated morbidity and mortality and pose a challenge to the treating physicians. Despite of lacking clinical data, the use of antifungal combination therapy is often considered to improve response rates in an immunocompromised patient population. We herein report a case of refractory invasive candidiasis in a liver transplant recipient treated successfully with a combination of isavuconazole und high-dose liposomal amphotericin B. The antimycotic combination treatment was able to clear a bloodstream infection with C. glabrata and led to regression of bilomas among tolerable side effects. The use of the above-mentioned antifungal combination therapy in a liver transplant recipient has not been reported previously. This case highlights the efficacy and safety of antifungal combination therapy in immunocompromised patients with refractory invasive candidiasis.

     

Infecc Ón Humana PorNocardia Asteroides,En México

Summary Although the existence ofNocardia asteroides as a human pathogen in Mexico has been known for several years, up to now there are only few reports in our country concerning the isolation of this microorganism both as a saprophyte as well as human or animal pathogen. This fact is more interesting when compared with the high incidence ofN. brasiliensis, causal agent of actinomycotic mycetoma, the most frequently observed deep mycosis in Mexico.In this paper, the isolation of eigth strains ofN. asteroides from clinical specimens is reported. Five strains were isolated from sputa, one from a fistulous lesion of the scrotum, one from a brain abscess and one more from a foot mycetoma.One of the strains isolated from sputum was in a case of pulmonary nocardiasis; another of these strains was isolated from a patient with coexistent pulmonary tuberculosis and nocardiasis. With the last three strains isolated from sputa, it was not possible to establish any correlation between the isolations and the clinical and radiological findings of the patients; two of the patients had pulmonary tuberculosis and the isolation ofN. asteroides was achieved only in one occasion in each case, with scant growth, and observed in routine cultures made for tubercle bacilli. The last strain was isolated from the sputum of a patient with chronic bronchitis. Some comments are made to the possibility that these three last isolations merely represent a saprophytic phase of the fungus.Some clinical and other features are discussed concerning the other three cases of scrotal fistula, brain abscess and mycetoma produced byN. asteroides.     

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.     

Age dependent alterations in phospholipid composition of a saprophytic and a pathogenic strain of Nocardia

Nocardia polychromogenes (saprophytic) and Nocardia asteroides (pathogenic) showed characteristic patterns in changes of cellular lipids during growth. Total lipids and total phospholipids decreased with the age of the culture in the saprophytic strain, whereas in the pathogenic strain total lipids increased throughout the culture period and the total phospholipids decreased in the late stationary phase. The decrease in total phospholipids in saprophytic strain was reflected in the individual phosphatides. In the pathogenic strain, the phosphatidylinositomannoside content doubled in early stationary phase. Differences were observed in fatty acid composition of phosphatides at various stages of growth, but the ratio of saturated to unsaturated fatty acids remained unaltered.     

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.     

Global Burden of Human Mycetoma: A Systematic Review and Meta-analysis

Mycetoma is a chronic infectious disease of the subcutaneous tissue with a high morbidity. This disease has been reported from countries between 30°N and 15°S since 1840 but the exact burden of disease is not known. It is currently unknown what the incidence, prevalence and the number of reported cases per year per country is. In order to estimate what the global burden of mycetoma is, a meta-analysis was performed. In total 50 studies were included, which resulted in a total of 8763 mycetoma cases. Most cases were found in men between 11 and 40 years of age. The foot was most commonly affected. Most cases were reported from Mexico, Sudan and India. Madurella mycetomatis was the most prevalent causative agent world-wide, followed by Actinomadura madurae, Streptomyces somaliensis, Actinomadura pelletieri, Nocardia brasiliensis and Nocardia asteroides. Although this study represents a first indication of the global burden on mycetoma, the actual burden is probably much higher. In this study only cases reported to literature could be used and most of these cases were found by searching archives from a single hospital in a single city of that country. By erecting (inter)national surveillance programs a more accurate estimation of the global burden on mycetoma can be obtained.     

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.     

The efficiency of 1M NH4Cl and 2 M NaCl for the isolation of pathogenic Nocardia from soil

The present paper reports an improvement to the classical method of the paraffin bait, by the usage of 1 M NH₄Cl or 2 M NaCl to eliminate contaminant microflora of soil.The purpose is to introduce a change in the paraffin bait method in order to reduce time required to isolate pathogenic strains of Nocardia from their natural sources.For this study three main criteria were used: a) Determination of the inhibitory effect of different concentrations of salts on soil microflora; b) The tolerance of Nocardia brasiliensis, Nocardia asteroides and Nocardia caviae (Nocardia otitidis caviarum) strains to these chemical inhibitors; c) Determination of the efficiency of salts in the isolation of Nocardia from soil when strains are grown on paraffin baits.