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Leishman (a) and Giemsa (b) stained peripheral blood thick smear showing ring stagePlasmodiumfalciparum. Parasites are easier identified in the Giemsa-stained slides.
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Microscopy of peripheral blood thin and thick films remains the reference for malaria diagnosis. Although Giemsa staining is most commonly used, the Leishman staining method provides better visualization of the nuclear chromatin pattern of cells. It is less well known whether accuracy of parasitaemia assessment is equally accurate with the latter m...
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... total of 276 pa- tients showed other abnormalities (Table 1). According to two independent slide readers, subjective comparison of the methods revealed that parasite identification was easier in thick smear preparation stained according to Giemsa (Figure 1). However, in the thin smears, slides stained according to Leishman had a better colour con- trast between the parasite cytoplasm and the red cell cytoplasm, in particular in case of mature trophozoite stage parasites (Figure 2). ...
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A flow cytometric (FACS) detection method for Plasmodium falciparum cultures (P. falciparum) was developed using SYBR Green I and CD235A and compared against the Giemsa stained microscopic examination. The cultured P. falciparum were spiked into red blood cells (RBCs) to yield parasitemia, ranging from 0.01% to 22.0%. FACS analysis demonstrated a c...
Citations
... Giemsa stains cytoplasm nicely, but the cell nucleus and cytoplasmic granules are stained lightly. 9 The LG cocktail, which combines these two stains, has the benefits of both the individual stains namely good nuclear morphology, fine nuclear and cytoplasmic contrast, cytoplasmic granule staining, and good metachromatic colour to the background material. 10 For diagnosis of malignancy, one of the most important criteria is its nuclear features. ...
FNAC is an outpatient procedure and has gained importance in mass screening programs. Staining procedure like MGG and Pap is time-consuming and needs trained persons. So there is a need for a staining procedure that is quick and easy to do. In this regard, LG cocktail has been suggested in many kinds of literature. LG cocktail is a combination of both Leishman and Giemsa stains, which has the advantage of both stains.
This is a prospective study at department of pathology, VSSIMSAR Burla. A total of 153 cases were studied from the cytology section. The slides were simultaneously stained with MGG and LG cocktail. The slides were viewed and scored independently by different pathologists. Quality Index (QI) was calculated by dividing this score by the maximum score possible.
Quality index of LG cocktail stain is 0.77 compared to MGG Quality Index 0.61. LG cocktail is better than MGG Stain overall staining quality, cytoplasmic and nuclear staining, and staining of background material.
LG cocktail has better QI than MGG stain. Thus the use of LG cocktail in cytological staining may increase the overall efficacy by saving time and decreasing the manpower need. These two advantages can help us in mass screening programs and its use in the crowded out-patient department.
... Malarial parasites are known to specifically invade red blood cells, multiplying within them in order to evade the immune response. The haematological profile of most infected patients is altered and typically includes severe anaemia, coagulation disturbances, leukocyte numerical or functional changes and spleen involvement to varying degrees [14], depending upon factors such as malarial endemicity and immunity, background haemoglobinopathy, demographic factors, nutritional status, etc [15,16]. Abnormalities reported are found to be most pronounced in P. falciparum infection, probably as a result of the higher levels of parasitemia found in these patients [17]. ...
Introduction: In Mumbai, an overpopulated metropolis with tropical conditions, malaria poses a persistent challenge, particularly in damp slum areas. Despite reported declines in death and incidence rates, the city faces a significant burden. Our project focuses on an unusual aspect: a heightened incidence among males, especially during sporadic monsoon outbreaks and in areas undergoing extensive redevelopment. Through an epidemiological survey of infected populations in municipal clinics, supported by microscopic, hematological, and microbiological evidence, we aim to provide insights into this atypical pattern. Materials and Methods: A cross-sectional, partly retrospective study was designed with participants being sourced from regional slums and chawls, during the immediately preceding three-year period of project completion. Patients attending OPDs were randomly selected from amongst those with febrile symptoms related to malarial disease and tested. Symptomatic individuals were tested for malaria microscopically using peripheral blood smear (PBS) and confirmed by employing the malaria antigen test (RMAT). Blood urea by enzymatic method and serum creatinine by Jaffes method. Results: Out of 1651 participants, 8.18% were malaria-positive, predominantly males (87.41%). P. vivax was the main parasite (87.41%), followed by P. falciparum (6.67%) and mixed infections (5.93%). ANOVA revealed significant RBC count differences (F = 6.32, p = 0.003). Tukey's HSD test showed P. vivax patients had higher RBC counts than mixed infection (p = 0.002), while P. falciparum counts were similar to P. vivax (p = 0.089) but higher than mixed infection (p = 0.014). Predominance of P. vivax emphasizes its impact on RBC counts and diagnostic complexities. The p values indicate that for all parameters, except serum creatinine in males, there is a statistically significant difference in the mean values between the malaria positive and negative groups Conclusion: This study highlights high malaria prevalence in urban India, with males more susceptible, possibly due to increased mosquito exposure. P. vivax is predominant, aligning with national data, emphasizing the need for tailored control measures. Surprisingly, P. vivax is associated with higher RBC counts than P. falciparum or mixed infection, suggesting differences in erythrocytic cycles. These findings have critical implications for regional malaria management, prompting further research into underlying mechanisms.
... A blood specimen collected from the patient was spread as a blood smear, stained with Giemsa stain, and examined with a 100X oil immersion objective. Classic ring-shaped/headphone-shaped trophozoites were observed in cases of Plasmodium Falciparum Malaria infection [21]. ...
Plasmodium falciparum is the parasite responsible for most malarial infections in Nigeria. Malaria is the most widespread parasitic infectious disease in the world. This study aimed to determine the effect of Plasmodium falciparum malaria infection on the amplitude of accommodation (AA) in young adults. The study was a prospective laboratory and clinical study carried out at the Madonna University Teaching Hospital Elele, Rivers State, Nigeria. Young adults between the ages of 18 and 35 years were included in this study. Informed consent was obtained from the subjects to participate in the study. To confirm the presence of Plasmodium falciparum malaria infection, preliminary tests including case history, external and internal eye examination, retinoscopy, and subjective refraction were performed to determine subjects who satisfied the inclusion and exclusion criteria. The amplitude of accommodation using the push-up method was measured just before treatment and two weeks after recovery from malaria infection. Results show that the mean AA during malaria infection was 7.25±1.73D and 9.03±1.72D after recovery; showing a mean difference of-1.78D (24.55%). This reduction in AA during malaria infection was found to be statistically significant (t(119) =-33.3, p(0.00)˂0.05) when tested using the paired sample t-test at a significance level of 0.05.
... The slides with blood smears were air-dried, and the dye was added dropwise onto them, allowing it to stain for 10 min. The stained slides were washed with tap water and air-dried again before microscopy examination [56]. ...
Background:
Advancements in nanoscience have led to a profound paradigm shift in the therapeutic applications of medicinally important natural drugs. The goal of this research is to develop a nano-natural product for efficient cancer treatment.
Methods and results:
For this purpose, mesoporous silica nanoparticles (MSNPs) were formulated, characterized, and loaded with caffeine to develop a targeted drug delivery system, i.e., caffeine-coated nanoparticles (CcNPs). In silico docking studies were conducted to examine the binding efficiency of the CcNPs with different apoptotic targets followed by in vitro and in vivo bioassays in respective animal models. Caffeine, administered both as a free drug and in nanomedicine form, along with doxorubicin, was delivered intravenously to a benzene-induced AML model. The anti-leukemic potential was assessed through hematological profiling, enzymatic biomarker analysis, and RT-PCR examination of genetic alterations in leukemia markers. Docking studies show strong inter-molecular interactions between CcNPs and apoptotic markers. In vitro analysis exhibits statistically significant antioxidant activity, whereas in vivo analysis exhibits normalization of the genetic expression of leukemia biomarkers STMN1 and S1009A, accompanied by the restoration of the hematological and morphological traits of leukemic blood cells in nanomedicine-treated rats. Likewise, a substantial improvement in hepatic and renal biomarkers is also observed. In addition to these findings, the nanomedicine successfully normalizes the elevated expression of GAPDH and mTOR induced by exposure to benzene. Further, the nanomedicine downregulates pro-survival components of the NF-kappa B pathway and upregulated P53 expression. Additionally, in the TRAIL pathway, it enhances the expression of pro-apoptotic players TRAIL and DR5 and downregulates the anti-apoptotic protein cFLIP.
Conclusions:
Our data suggest that MSNPs loaded with caffeine, i.e., CcNP/nanomedicine, can potentially inhibit transformed cell proliferation and induce pro-apoptotic TRAIL machinery to counter benzene-induced leukemia. These results render our nanomedicine as a potentially excellent therapeutic agent against AML.
... The original Romanowsky method was modified by William Boog Leishman a British pathologist and was called as Leishman's stain. 1 The advantage of Leishman stain over Giemsa staining (another Romanowsky stain used in hematology) is that it can be prepared easily, low cost and staining can be done within 10-15 minutes. 2,3,4 Few modifications in preparation of Leishman stain has been attempted that can aid in reducing the duration of staining of peripheral blood smears. One such attempt is addition of phenol, an accentuating agent that helps to reduce the duration of staining when added to Leishman stain. ...
Context: Leishman stain, one of the Romanowsky stain, is used as the stain of choice for routine staining of peripheral blood films since many years in most of the hematology laboratories worldwide. But it has a disadvantage of consuming around 15-20 minutes of time for staining alone, which can lead to increase in the turnaround time for reporting peripheral blood smears. In this study Phenol, an accentuating agent is used as the vehicle of innovation for preparing modified Leishman stain that can aid to reduce the staining time without altering the quality of smears. Aim: To know the optimal phenol: Leishman powder ratio that is appropriate for preparing modified Leishman stain and to evaluate the staining efficacy of this modified stain over the conventional Leishman by a scoring system. Settings and Design: It is a cross sectional study done after taking the ethical clearance from Institutional Ethical Committee. Methods and Material: The study was conducted in the department of clinical Hematology Laboratory of a tertiary health care centre in South India from July 2019 to August 2019, on a total of 600 smears prepared from EDTA anticoagulant blood samples that were stained by both conventional as well Modified Leishman stain. These smears were then given a score based on five parameters and quality index was calculated. Statistical Analysis: The results obtained were entered into a Microsoft excel sheet. The minimum and maximum scores for a single case ranged from 0-10. Quality index of these stains was obtained by dividing the actual score by the maximum score possible. No statistical instruments were used as the study was based on interpretation of staining characteristics. Results: Modified Leishman stain at phenol: Leishman powder ratio of 1:5 on day 10 gave best staining quality when compared to other days at various concentrations with a quality index of 0.86 and was as good as conventional Leishman stain Conclusions: Modified Leishman Stain prepared by adding phenol to conventional Leishman stain can give better staining results in 4 minutes unlike the conventional Leishman stain which takes 15-20 minutes of time for staining peripheral blood smears.
... The dried thick blood smears were prepared in accordance with a method described by Ref. [31]. The "plus subsystem" was used to quantify the malaria parasites in accordance with centre for disease control criteria [32]. ...
Background: Paediatric febrile illnesses pose diagnostic challenges in low-income countries. Western Uganda is endemic for both malaria and typhoid but the true prevalence of each individual disease, their co-infections and associated factors are poorly quantified.
Objective: To determine the prevalence of malaria, typhoid, their co-infection, and associated factors amongst febrile children attending the paediatrics and child health department of Kampala International University Teaching Hospital (KIU-TH) in Western Uganda.
Methods: Cross-sectional study used a survey questionnaire covering demographics, clinical and behavioural variables. We obtained blood for peripheral films for malaria and cultures for typhoid respectively; from 108 consecutively consented participants. Ethical approval was obtained from KIU-TH research and ethics committee (No. UG-REC-023/201,834). Multivariate regression analysis was performed using Stata 14.0 (StataCorp. 2015) at 95% confidence interval, regarding p < 0.05 as statistically significant.
Results: Majority of participants were males 62% (n = 67), cared for by their mothers 86.1% (n =93). The prevalence of malaria was 25% (n = 27). The prevalence of typhoid was 3.7% (n = 4), whereas the prevalence of malaria-typhoid co-infection was 2.8% (n = 3). Using treated water from protected public taps was associated with low malaria-typhoid co-infection [p = 0.04; aOR = 0.05, 95%CI [0.003–0.87], whereas drinking unboiled water from open wells increased the risk for the co-infection [p = 0.037, cOR = 17, 95%CI (1.19–243.25)].
Conclusions: The prevalence of blood culture confirmed malaria-typhoid co-infection in children was lower than previously reported in serological studies. These findings emphasize the need to use gold standard diagnostic investigations in epidemiological studies. Educational campaigns should focus on the use of safe water, hygienic hand washing, and proper waste disposal; and should target mothers who mainly take care of these children.
... Although the Leishman staining method is less widely used than the Giemsa staining method, it offers better visualizations and sensitivity while taking less time than the Giemsa staining method to detect malaria parasites [26][27][28]. In addition, studies like [29][30][31] have used Leishman staining to identify malaria parasites in blood smears. ...
Malaria belongs to the class of the deadliest infectious diseases in the world. The generally available tools to diagnose this disease, the microscopy and rapid diagnostic test (RDT), have many limitations. Alternative diagnostic techniques with superior results are inaccessible to developing countries with more prevalent cases. Early detection of the infection is critical. Computer-assisted methods are needed. This study surveys the performance of the computer-assisted techniques used in malaria diagnosis and the preprocessing techniques to render the data usable. The survey illustrates, compares and discusses computer-assisted methods results, considering different performance metrics. It highlights how artificial intelligence can strengthen the fight against disease.KeywordsMalariaDiagnosisMachine learningComputer-assisted techniquesPerformance metricsDeep learning
... Staining of smears was carried out by the method of rapid staining with Leukodif-200 using the V-Chromer ® III device (West Medica, Austria, 2019). Light microscopy was performed on a Kern Obe digital microscope (Kern, Germany, 2021) (Satpathy et al., 2014). The intensity of parasitaemia was calculated by the percentage of affected erythrocytes in 10 fields of view. ...
With babesiosis in dogs, the capillary network is especially strongly affected due to the stimulation of the hemostasis system and the development of thrombus formation in the microvasculature. Acute renal failure and even death of animals are caused by shock phenomena and the syndrome of disseminated intravascular coagulation. Clinical, laboratory (hematological, biochemical, coagulometric, hemodynamic, enzyme immunoassay) methods of research were conducted. Dogs with babesiosis have fever, anemia, and hemoglobinuria, hematologically established anemia, systemic inflammation, thrombotic condition. Hemodynamic parameters determine the state of shock, which in dynamics becomes decompensated and irreversible. The appearance of poikilocytes in the blood of sick dogs indicates significant metabolic disorders in their body. The coagulogram reveals the syndrome of disseminated intravascular coagulation. Syndrome markers significantly changed with a trend to increase during nephropathy. In addition, uremia, hyperuricemia, hyperkalemia, hyponatremia, hyperphosphatemia, creatininemia were found in the sick dogs, which are reliable indicators and determine the development of an acute form of glomerulonephritis and progressive acute renal failure. According to the study of the general biochemical status in animals, hepatopathy in the form of acute hepatitis and pancreatitis, and general intoxication syndrome were established. In our findings: during acute spontaneous babesiosis, dogs developed nephropathy, which has two clinical stages: hemoglobinuria and oliguria/anuria. The pathogenetic basis for the development of glomerulonephritis and acute renal failure is the syndrome of disseminated intravascular coagulation in response to the influence of a pathogen, which causes shock, forming a vicious circle of shock. Under such conditions, renal failure increases and eventually leads to the death of the animal. The studied data will contribute to the expansion of the conceptual understanding of the pathogenesis of canine babesiosis and determine the leading role of complications in the development of kidney failure.
... This was in line with research by Rinny et al., (2018) that Giemsa is very good at identifying various granulocyte cells and other blood cells, producing a clear core picture, differentiating the basophilic or eosinophilic components of lymphoid and myeloid cells [12]. The main advantage of Giemsa is more durable in tropical climates and excellent to research blood parasites [13]. ...
Giemsa stain is one of the synthetic stains consisting of eosin and blue methylene that are useful for staining cells. However, the use of synthetic stain turns out to have several drawbacks, some of which are not eco-friendly, quite expensive and harmful to health. Therefore, there was a need of alternative coloring that is natural, more eco-friendly, inexpensive, and especially not harmful to health. This study used alternative dye, jamblang ethanol extract (Syzigium cumini), containing anthocyanins. Anthocyanins are a class of organic chemical compounds that could dissolve in polar solvents, and were responsible for giving orange, red, purple, blue, and black colors in higher plants such as flowers, fruits, seeds, vegetables and tubers. The results of this study indicated that Giemsa stain contained eosin and blue methylene so that while in use, it could produce blood cells such as erythrocytes, leukocytes and platelets whose appearance was clear under a microscope. Meanwhile, jamblang ethanol extract stain contains acidic anthocyanins so that this staining can only stain blood cells in the form of erythrocytes, if the leukocytes and platelets could not be stained properly due to leukocytes and platelets having acidic cell nuclei. Therefore, it can be concluded that the staning of Giemsa has a better quality than that of jamblang ethanol extract, but jamblang extract can be used as an alternative stain from Giemsa to indicate erythrocyte cells.
... However, considering the diagnosis of erythrocytes in a blood smear, the giemsa stain proved to be much superior to safranin. In a comparative study of Leishman and giemsa staining, Sathpathi et al. (2014) suggested the superiority of giemsa staining for a thick blood smear. In the present study, the giemsa staining produced distinct and evident erythrocytic morphology with subsequent alterations and abnormalities. ...
Tilapia is one of the most consumed farmed fish, which requires the use of antibiotics in certain phases of its production. This study assessed the safety of 30 days of oral florfenicol (FFC) dosing at 0–10 times the therapeutic dose (1 × : 10 mg/kg biomass/day) in Oreochromis niloticus juveniles. Behavioural changes, feed consumption, mortality and biomass were evaluated. Besides, the levels of serum glucose, calcium, chloride, creatinine, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and blood cell morphology were determined at scheduled intervals. The 30 days of oral FFC dosing caused 3.33% (1 ×) to 18.33% (10 ×) mortalities, reduced feed intake and biomass in a dose-dependent manner. The fish fed the therapeutic dose recorded 1.25-fold increase in biomass, while the control group recorded 1.45-fold increase in 30 days. No significant erythrocyte morphological alterations were observed in the 1 × group compared to the control. However, marked morphological alterations like tear-shaped, spindle-shaped and degenerative erythrocytes in higher dosing groups indicated FFC cytotoxicity. All the serum biomarkers of O. niloticus increased significantly on day 10 and day 30 FFC dosing in a dose-dependent manner, except for calcium and chloride, which reduced significantly during the dosing period. Within 2 weeks of suspension of FFC dosing, the serum biomarker levels became normal except for alkaline phosphatase and creatinine. The recovery of biomass, feed intake, serum biomarker levels and erythrocyte morphological changes suggested that the FFC-induced changes are reversible. This study has, thus, proclaimed the safety of FFC at the therapeutic dose in O. niloticus.
