Bone Marrow Aspirate Culture Research Articles

Invasive fungal infection by Cryptococcus neoformans var. grubii with bone marrow and meningeal involvement in a HIV-infected patient: a case report

BackgroundCryptococcosis is a common opportunistic infection in patients infected by Human Immunodeficiency Virus (HIV) and is the second leading cause of mortality in Acquired Immunodeficiency Syndrome (AIDS) patients worldwide. The most frequent presentation of cryptococcal infection is subacute meningitis, especially in patients with a CD4+ T Lymphocytes count below 100 cells/μL. However, in severely immunosuppressed individuals Cryptococcus neoformans can infect virtually any human organ, including the bone marrow, which is a rare presentation of cryptococcosis.Case presentationA 45-year-old HIV-infected male patient with a CD4+ T lymphocyte count of 26 cells/μL who presented to the emergency department with fever and pancytopenia. Throughout the diagnostic evaluation, the bone marrow aspirate culture yielded encapsulated yeasts in budding, identified as Cryptococcus sp. The bone marrow biopsy revealed a hypocellularity for age and absence of fibrosis. It was observed presence of loosely formed granuloma composed of multinucleated giant cells encompassing rounded yeast like organisms stained with mucicarmine, compatible with Cryptococcus sp. Then, the patient underwent a lumbar puncture to investigate meningitis, although he had no neurological symptoms and neurological examination was normal. The cerebrospinal fluid culture yielded Cryptococcus sp. The species and genotype identification step showed the infection was caused by Cryptococcus neoformans var. grubii (genotype VNI). The patient was initially treated with amphotericin B deoxycholate plus fluconazole for disseminated cryptococcosis, according to guideline recommendations. However, the patient developed acute kidney injury and the treatment was switched for fluconazole monotherapy. The symptoms disappeared completely with recovery of white blood cells and platelets counts. Cerebrospinal fluid cultures for fungi at one and two-weeks of treatment were negative.ConclusionsBone marrow infection caused by Cryptococcus neoformans is a rare presentation of cryptococcosis. The cryptococcal infection should be included for differential diagnosis in HIV-infected patients with fever and cytopenias, especially when CD4+ T lymphocytes count is below 100 cells/μL.

Cell-secreted extracellular matrix influences cellular composition sequestered from unprocessed bone marrow aspirate for osteogenic grafts.

Bone marrow aspirates provide a rich source of cells for use in tissue engineering of bone and other clinical indications. However, progenitor cells such as mesenchymal stem cells (MSCs) account for a small fraction of nucleated cells in bone marrow aspirate (BMA), requiring extensive culture expansion. Accessory cell populations such as endothelial or hematopoietic cells can potentiate the bone-forming potential of MSCs, and cell-secreted extracellular matrix (ECM) can increase cell seeding efficiency and osteogenic differentiation of heterogeneous cell populations. In this study, we hypothesized that cell-secreted ECM could be used to sequester MSCs and accessory cells from BMA for bone regeneration. To generate 3D implantable constructs, BMA was resuspended in media with or without type I collagen or ECM and injected into a perfusion bioreactor system. The addition of protein coatings increased cell seeding efficiency compared to uncoated scaffolds. Compared to fresh BMA, the culture of BMA on all scaffolds reduced the proportion of CD45+ myeloid cells and increased CD31+CD45- endothelial cells. Compared to uncoated scaffolds, we observed a 143- and 30-fold increase in MSCs when fresh BMA was cultured on ECM- or collagen-coated scaffolds, respectively. Upon subcutaneous implantation, ECM-coated scaffolds promoted cell survival and early vascularization. However, bone formation was comparable across all implant groups, suggesting additional osteogenic cues are necessary to increase the bone forming potential of fresh BMA. These results motivate further investigation into strategies which elicit more robust bone regeneration by tissue aspirates.

Bone marrow granuloma in typhoid Fever: a morphological approach and literature review.

Typhoid fever is one of the few bacterial infections in humans where bone marrow evaluation is routinely recommended. However, the morphological aspect of typhoid fever in bone marrow has been rarely described in the literature. We describe a 25-year-old male patient who presented with prolonged fever suspected to be of tubercular etiology. Bone marrow examination showed well-formed histiocytic and epithelioid granulomas and erythrophagocytosis; and the bone marrow aspirate culture grew Salmonella typhi A. In view of potential clinical implications, typhoid fever should be considered as a differential diagnosis to tuberculosis in the evaluation of prolonged fever; especially in high prevalent areas. We suggest that erythrophagocytosis may serve as a morphological marker in typhoid granulomas in the bone marrow; and bone marrow culture should be submitted in every suspected case for appropriate patient management.

Counts and cards – a novel way to detect typhoid infections in the ED

Objective The diagnosis of typhoid fever is made by growth of the causative microorganism from culture of bone marrow aspirate or blood which is time dependent. In a busy ED like ours, typhoid fever is a frequent presentation. There is need for a rapid and reliable test made available at the bed side, to detect typhoid infections. The sensitivity and specificity of Enterocheck WB, a card test that detects IgM antibodies to salmonella typhi and low eosinophil counts were compared individually and together, with the gold standard of blood culture using BacT/Alert.

Identification of mesenchymal stem cells and osteogenic factors in bone marrow aspirate and peripheral blood for spinal fusion by flow cytometry and proteomic analysis.

BackgroundAn in vivo animal study and a prospective clinical study have indicated that bone marrow aspirate (BMA) augments spinal arthrodesis. However, there is no quantified data to explain why fusion rate can be augmented by BMA in lumbar posterolateral fusion.MethodsTo analyze the proportion of mesenchymal stem cells (MSCs) and osteogenic factors in human BMA and peripheral blood (PB) of the same patient. Autologous BMA and PB from the patients were analyzed by flow cytometry (FACS) using cell markers for MSCs. The osteogenic potential of MSCs was determined by alkaline phosphatase (ALP) activity and calcium level quantification. Proteomics were used for the qualitative and quantitative mapping of the whole proteome from BMA and PB plasma. The mass-to-charge ratio was calculated by time-of-flight mass spectrometry (TOF-MS). The overexpression of protein was confirmed using Western blot analysis.ResultsThe proportion of MSCs (CD34−/CD29+/CD105+) was higher in the BMA than that in the PB. Colony-forming cell (CFC) assays suggested that fewer colonies were formed in PB cultures than in BMA culture. There was no significant difference in the osteogenic potential of the MSCs between the PB and BMA. Proteomic mass spectrometry assays suggested that the levels of catalase (osteoclast inhibitor) and glutathione peroxidase 3 (osteogenic biomarker) were higher in the BMA than those in the PB, and this was confirmed by Western blot analysis.ConclusionsThe proportions of MSCs and osteogenic factors were higher in the BMA than in the PB. This may explain why fusion rate can be augmented by BMA in lumbar posterolateral fusion.

Molecular diagnosis of canine visceral leishmaniasis: A comparative study of three methods using skin and spleen from dogs with natural Leishmania infantum infection

Polymerase chain reaction (PCR) and its variations represent highly sensitive and specific methods for Leishmania DNA detection and subsequent canine visceral leishmaniasis (CVL) diagnosis. The aim of this work was to compare three different molecular diagnosis techniques (conventional PCR [cPCR], seminested PCR [snPCR], and quantitative PCR [qPCR]) in samples of skin and spleen from 60 seropositive dogs by immunofluorescence antibody test and enzyme-linked immunosorbent assay. Parasitological analysis was conducted by culture of bone marrow aspirate and optical microscopic assessment of ear skin and spleen samples stained with Giemsa, the standard tests for CVL diagnosis. The primers L150/L152 and LINR4/LIN17/LIN19 were used to amplify the conserved region of the Leishmania kDNA minicircle in the cPCR, and snPCR and qPCR were performed using the DNA polymerase gene (DNA pol α) primers from Leishmania infantum. The parasitological analysis revealed parasites in 61.7% of the samples. Sensitivities were 89.2%, 86.5%, and 97.3% in the skin and 81.1%, 94.6%, and 100.0% in spleen samples used for cPCR, snPCR, and qPCR, respectively. We demonstrated that the qPCR method was the best technique to detect L. infantum in both skin and spleen samples. However, we recommend the use of skin due to the high sensitivity and sampling being less invasive.

Parasitic Infections in Solid Organ Transplant Recipients

Parasitic Infections in Solid Organ Transplant Recipients

Clinical and laboratory features of disseminated histoplasmosis in HIV patients from Brazil

To identify the main clinical and laboratory features of disseminated histoplasmosis (DH) in human immunodeficiency virus (HIV) patients and compare them with those of HIV patients with other opportunistic diseases. Retrospective study of HIV patients comparing the clinical and laboratory data of patients with and without DH. Univariate and multivariate analyses were performed to verify the risk factors related to DH. In total, 378 HIV patients were included, 164 with DH and 214 with other opportunistic diseases. Acute renal failure, respiratory insufficiency and septic shock were more frequent in DH patients, who also had a higher mortality (32%vs. 14%, P < 0.001). Independent risk factors for DH were: acute renal failure [odds ratio (OR) 5.2; 95% confidence interval (CI) 3.2-8.5; P < 0.001], splenomegaly (OR 3.4; 95% CI 1.19-9.9; P < 0.001), respiratory insufficiency (OR 2.7 95% CI 1.5-5.0; P < 0.001), proteinuria (OR 2.7; 95% CI 1.3-5.2; P = 0.03), hypotension (OR 2.5; 95% CI 1.2-5.0; P = 0.008), hepatomegaly (OR 2.4; 95% CI 1.2-4.8; P = 0.01), cutaneous lesions (OR, 1.9; 95% CI 1.0-3.3; P = 0.02) and weight loss (OR 1.8; 95% CI 1.0-3.1; P = 0.03). Our results suggest that DH is a severe opportunistic disease with high mortality rate, which should be promptly recognized in order to provide early specific treatment.

Cord Blood-Derived Plasma Factor (CBPF) Potentiates the Low Cytokinetic and Immunokinetic Profile of Bone Marrow Cells in Pesticide Victims Suffering from Acquired Aplastic Anaemia (AAA): An in vitro Correlate

Cases of Acquired Aplastic Anaemia (AAA) in patients with a long history of pesticide exposure from agricultural fields have been investigated in our laboratory using an immunological approach. These patients showed moderate to severe degrees of bone marrow aplasia as a result of 9–12 years protracted exposure to pesticides which were mainly comprised of organophosphorous and organochloride compounds. The bone marrow aspirate culture was found to be severely deficient both in terms of differentiation and proliferation, and cell mediated immune function (CMI). We attempted ex vivo manipulation of the bone marrow population of patients in two different protocols: in one, stem cell factor (SCF), interleukin-3 (IL-3), and granulocyte-colony stimulating factor (G-CSF) were administered and, in the second set, cord blood-derived plasma factors (CBPF) were supplemented to evaluate the effects, if any. Simultaneously, two control groups including one for healthy normal control (N) and the second, for non-pesticide induced aplastic anaemia group of patients (NPAA) was also investigated for all the above parameters. Active colony formation and improved cellular immune activity (CMI) was observed more frequently in the CBPF treated group rather than that in the cytokine treated group. Surprisingly, administration of cytokines in the first set and CBPF in the second set triggered CD34 + cell generation as revealed through flow cytometric analysis (FACS). The effect was more pronounced in the second set. Investigations carried out with NPAA showed relatively insignificant effects with both cytokine and CBPF set up. The investigations indicated that AAA as induced by pesticides could be therapeutically manipulated by exogenous cytokines and growth factors and, more efficiently, by CBPF by way of immunopotentiation through microenvironmental supplementation.

Two cases of salmonella splenic abscess

Splenic abscess is a rare clinical entity with an incidence of 0.2 to 0.7 percent in autopsy based studies [1]. Less than 500 cases have been reported in world literature. Clinical presentation is non-specific and it is a potentially serious surgical problem associated with high mortality. The treatment has been a combination of total splenectomy or percutaneous drainage and appropriate antibiotic therapy. We report 2 cases who developed multiple splenic abscesses and were successfully treated. Case 1 A 21 year old man presented with high-grade intermittent fever with chills and rigors and productive cough of 10 days duration. On examination he had fever, left sided pleural rub and a palpable liver, 2cms below the right costal margin. Investigation showed a Hb of 13gm/dl; TLC 6900/cmm; DLC N48 L48 E03M02; blood sugar 71 mg/dl; bilirubin 0.9 mg/dl, SGOT 76 U/L, SGPT 56 U/L; blood urea 30 mg/dl and serum creatinine 1.3 mg/dl. Peripheral blood smear revealed leucocytosis with shift to left and toxic granules. Chest radiograph showed elevated left hemi-diaphragm. Lung parenchyma and costophrenic angles were normal. Abdominal ultrasonography (USG) showed splenomegaly with multiple hypoechoic areas measuring few mm to 6 cms in diameter. The Widal test was negative. Blood culture and bone marrow aspiration culture were sterile. Contrast enhanced computerized tomography of abdomen confirmed multiple hypodense lesions suggestive of splenic abscess. Injection ceftriaxone and amikacin were instituted. A USG guided percutaneous aspiration was done and 400ml of haemorrhagic pus was drained. Ziehl Neelsen and gram's stain were negative for bacteria. Pus culture grew Salmonella typhi. The fever persisted inspite of antibiotics and patient developed hypotension. A surgical consultation was sought and he underwent emergent splenectomy. The fever subsided from 3rd postoperative day and the general condition improved. Antibiotics were continued for 4 weeks. Sickling and HIV test were negative.

Detecting disseminated Mycobacterium avium complex infections in HIV-positive patients. The usefulness of bone marrow trephine biopsy specimens, aspirate cultures, and blood cultures.

Disseminated Mycobacterium avium complex (MAC) infections are common in patients with acquired immunodeficiency syndrome (AIDS). These patients frequently seek care with fever accompanied by generalized systemic symptoms and undergo bone marrow biopsy. It is our practice to stain all bone marrow trephine biopsy specimens from patients infected with HIV for acid-fast bacilli (AFB). We evaluated this practice by comparing the sensitivity and turnaround time for detection of MAC by biopsy specimen staining, bone marrow aspirate culture, and blood culture. Bone marrow trephine biopsy specimens with corresponding bone marrow aspirate and blood cultures from 86 HIV-positive patients were reviewed. Of the 86 patients, 30 had positive results for disseminated MAC infection, and all 30 of those patients had positive blood cultures. Bone marrow aspirate cultures identified 17 MAC-positive cases, and AFB staining of the biopsy specimen identified 9. The mean times to detection of MAC positivity were 1.1 days for AFB staining of the biopsy specimen, 19 days for bone marrow aspirate culture, and 16 days for blood culture. While AFB staining of biopsy specimens was the least sensitive of the detection methods, it was useful for the rapid diagnosis of disseminated MAC infection, allowing for prompt initiation of antimycobacterial therapy in one third of patients.

Diffuse cutaneous leishmaniasis with visceral dissemination in an AIDS patient in Guadeloupe, West Indies

Barton, S. E.; Davies, S.; Schroeder, K.; Arthur, C.; Cazzard, B. C. Author Information

Diagnostic value of a single, pre-treatment Widal test in suspected enteric fever cases in the Philippines

101 patients with a clinical suspicion of typhoid or paratyphoid (enteric) fever admitted to San Lazaro Hospital, Manila, Philippines, were studied by bacteriological culture of blood, rectal swab, urine and duodenal string capsule; 35 also had bacteriological culture of bone marrow aspirate. 44 of the patients were culture-confirmed as having enteric fever; the remainder were classified as non-enteric fever cases. Analysis of the pretreatment Widal agglutination titres of all patients revealed that using as a diagnostic criterion an antibody titre of ⩾ 1:80 to the O antigen of Salmonella typhi yielded a test specificity of 100%, although the corresponding sensitivity was only 64%. The sensitivity of the test could be increased to 80% by using different cut-off values for titres to flagellar antigens, but this concomitantly decreased the test specificity from 100 to 82%. The data indicate that a single pretreatment Widal test in suspected enteric fever cases is of definite diagnostic value, but that the results must be interpreted with caution and foreknowledge of the test's shortcomings and limitations.

Duodenal string-capsule culture compared with bone-marrow, blood, and rectal-swab cultures for diagnosing typhoid and paratyphoid fever.

The sensitivity of duodenal string-capsule culture (DSCC) was compared to that of bone-marrow-aspirate culture (BMAC), single 3-ml blood culture (BC), and rectal-swab culture (RSC) for isolating Salmonella typhi and Salmonella paratyphi type A from patients with typhoid and paratyphoid fever. In 36 of 154 patients DSCC could not be used, usually because the patient was too ill to swallow the capsule. In the remaining 118 patients DSCC was positive in 57.6%, RSC in 35.6%, BC in 54.2%, and BMAC in 85.6%. The sensitivity of DSCC was improved by an additional 4.7% if subcultured daily for seven days. The DSCC has no advantage over the combination of RSC and BC and is inferior in sensitivity to the BMAC. However, when a BMAC cannot be obtained, the addition of the DSCC to BC and RSC can be expected to improve the isolation rate by greater than 17%, to at least 85%.

Bone marrow cultures in patients infected with ‘anonymous’ mycobacteria

Summary Cultures for acid-fast bacteria were made from biopsies of the bone marrow n of 57 patients in whom such organisms had been grown from sputum. In four the results were inconclusive. Of 18 patients in whom the final diagnosis was lung infection with anonymous mycobacteria, eight gave positive cultures. Of 23 in whom definite lesions of the lungs produced by other causes were present, but where it was thought that anonymous mycobacteria might have been present as a complication, four gave positive cultures. Of 12 patients with pathological changes of the lung in whom anonymous mycobacterial infection was considered unlikely, one gave a positive culture. In 22 control patients thought at the time of examination to be suffering from haematological disorders, one man of 75 who was dying from cancer of the prostate and urinary infection gave a positive culture, possibly Myco. smegmatis. Culture of bone-marrow aspirate may at times be helpful in the diagnosis of infection with anonymous mycobacteria.