Blood Film Examination Research Articles

An Overview Study of Malaria Infection in Almaza Military Fever Hospital; An Egyptian Pilot Study

Background and study aim: Malaria is a life-threatening disease caused by parasites that are transmitted to people through the bites of infected female Anopheles mosquitoes.The aim of this study is to study the clinical presentations and outcomes of malarial infected cases attending Almaza Military Fever Hospital in Cairo. Patients and Methods: Fifty patients with malarial infection were selected from those admitted to Almaza Military Fever Hospital. The following investigations were done for all cases; (CBC), liver and renal function tests, serological tests (rapid diagnostic test for serum malarial antigens & microscopic examination of peripheral blood film) and abdominal US. Results: The majority of cases (76%) was already diagnosed and was coming from Peace Keeping Mission Forces in Africa. Congo was the most malaria-infected place (36%), then Ivory Coast (26%). Most of cases (80%) had intermittent fever. Six patients (12%) were admitted at ICU. The thick film method was the most sensitive diagnostic test (98%). P. falciparum was the commonest species among cases (80%) then P. ovale (20%). The best response in studied cases was poly-therapy (84%) while mono-therapy was effective in only 5 patients (10%), (82%) of cases were cured, one patient died and one patient had a relapse while 2 patient (4%) had recurred. Conclusion: Thick film is the most sensitive and informative test among diagnostic test modalities. Combined therapy (polytherapy) is preferable than monotherapy.

Clinico-hematological analysis of pancytopenia in adults – A two year prospective study

Background : Pancytopenia is a common hematological finding resulting from various diseases which requires thorough clinical history, physical examination and blood investigations so as to evaluate the cause and plan the management of pancytopenic patients. The aim of this study is to determine the incidence, various causes, common clinical presentations of pancytopenia and to correlate hematological parameters with clinical findings in differentiating causes of pancytopenia. Methods : A two year prospective study which included patients of age 15 years and above having pancytopenia on blood film examination ( Hb < 10g/dl, TLC < 4000/cmm and platelets < 1lakh/ cmm). Total 226 cases of pancytopenia were evaluated clinically along with hematological parameters and bone marrow aspiration in Central Laboratory of Department of Pathology, Tertiary care hospital, Navi Mumbai. Results : The most common causes of pancytopenia were malaria (50%), megaloblastic anemia (18.6%) and dengue (18.1%) followed by hypersplenism (7.1%) and iron deficiency anemia. The other uncommon causes were septicemia (2.2%), AIDS (1.8%), tuberculosis (1.8%), aplastic anemia (1.3%), leptospirosis (0.9%), dimorphic anemia (0.9%), dyskeratosis congenita (0.4%) and myelodysplastic syndrome (0.4%). The age of the patients ranged from 15-85 years with male preponderance. The most common clinical features were weakness (80%), fever (72.1%), pallor (100%) and hepatomegaly (62.8%). Normocytic normochromic was the predominant blood picture. Conclusion : The present study concludes that detailed primary hematological investigations and other supportive biochemical investigations can be helpful to rule out or to diagnose causes of pancytopenia and invasive procedures like bone marrow aspiration or biopsy can be avoided in majority of cases. DOI:10.21276/APALM.1519

Diagnosis of Atrial Fibrillation in a Horse in Association with Microfilariasis

A 12-year-old crossbred mare was presented to the clinic with a history of frequent reduction in the working capacity. Clinical examination revealed congested mucus membranes, elevated respiratory rate and arrhythmia. Laboratory analysis revealed the anemia, lowered packed cell volume, lowered albumin, total protein, sodium and potassium levels. Electrocardiography confirmed the presence of the atrial fibrillation and wet blood film examination showed the presence of the motile microfilaria organisms. The horse was treated with oral administration of quinidine sulfate and other supportive therapy.

Microfilaria in fine-needle aspiration cytology of renal mass – an unusual site: A rare case report

In tropical countries like India, filariasis is one among the major public health problems. Most commonly (90%) cases of lymphatic filariasis caused by wuchereria bancrofti, and rest (10%) cases by Brugia malayi. In India, filariasis is transmitted by the bite of culex or mansonia mosquito species. The diagnostic method of choice in filariasis is the detection of microfilaria in peripheral blood film examinations. Microfilaria was incidentally detected in fine-needle aspiration cytology (FNAC) smears from different tissues, organs, and serous cavities. However, the presence of a filarial worm in renal mass aspirate is unusual, and no literature has been published with best of our knowledge. We report a case in which ultrasound-guided FNAC renal mass revealed the presence of microfilaria in the background of renal cell carcinoma cells.

The identification of Theileria bicornis in captive rhinoceros in Australia.

Poaching of both black (Diceros bicornis) and white (Ceratotherium simum) rhinoceros in Africa has increased significantly in recent years. In an effort to ensure the survival of these critically endangered species, breeding programs were established in the 1990s in Australia, where a similar climate and habitat is available. In this study we examined blood samples from two C. simum, including a 16 yr old female (Aluka) who died in captivity, and a 17 yr old asymptomatic male (Umfana). Bloods from seven healthy D. bicornis housed at the zoo were also collected. All samples were tested for the presence of piroplasms via blood smear and PCR. A generic PCR for the 18S rRNA gene of the Piroplasmida revealed the presence of piroplasm infection in both dead and asymptomatic C. simum. Subsequent sequencing of these amplicons revealed the presence of Theileria bicornis. Blood smear indicated that this organism was present at low abundance in both affected and asymptomatic individuals and was not linked to the C. simum mortality. T. bicornis was also detected in the D. bicornis population (n = 7) housed at Taronga Western Plains Zoo using PCR and blood film examination; however only animals imported from Africa (n = 1) tested T. bicornis positive, while captive-born animals bred within Australia (n = 6) tested negative suggesting that transmission within the herd was unlikely. Phylogenetic analysis of the full length T. bicornis 18S rRNA genes classified this organism outside the clade of the transforming and non-transforming Theileria with a new haplotype, H4, identified from D. bicornis. This study revealed the presence of Theileria bicornis in Australian captive populations of both C. simum and D. bicornis and a new haplotype of the parasite was identified.

Bleeding is not the main clinical issue in many patients with inherited thrombocytopaenias.

Bleeding diathesis has been considered for a long time the main clinical issue impacting the lives of patients affected by inherited thrombocytopaenias. However, the number of known inherited thrombocytopaenias greatly increased in recent years, and careful evaluation of hundreds of patients affected by these 'new' disorders revealed that most of them are at risk of developing additional life-threatening disorders during childhood or adult life. These additional disorders are usually more serious and dangerous than low platelet count. For instance, it is known that mutations in RUNX1, ANKRD26 and ETV6 cause congenital thrombocytopaenia, but we now know that they also predispose to haematological malignancies. Similarly, MYH9 mutations result in congenital thrombocytopaenia and increase the risk of developing kidney failure, cataracts and hearing loss at a later stage, while MPL mutations cause a congenital thrombocytopaenia that almost always evolves into deadly bone marrow failure. Thus, identification of patients with these disorders is essential for evaluation of their prognosis, enabling effective genetic counselling, personalizing follow-up and giving appropriate treatments in case of development of additional diseases. Careful clinical evaluation and peripheral blood film examination are extremely useful tools in guiding the diagnostic process and identifying the candidate genes to be sequenced.

Hereditary Haemoglobin Disorders of Anaemic Patients Attending in a Rural Tertiary Level Hospital

Background: Regularly patients attending in the hospital with severe illness for medical treatment. A large number of patients came with features of anaemia. Many of them show hereditary haemoglobin disorders.Objective: This study was done to find out the pattern of hereditary haemoglobin disorders of anaemic patients attending in the hospital for medical treatment in a tertiary level rural hospital.Method: This descriptive type of retrospective study was performed with 151 cases of anaemic patients whose whole blood show abnormality in haemolobin on capillary haemoglobin electrophoresis. Haemoglobin as well as haematocrit and red blood cells indices were performed by using fully automated haematology lab automation (Sysmex XN-1000). Then haemoglobin electrophoresis was performed to see haemoglobin disorder by using Capillary-2 Haemoglobin Electrophoresis, Sebia, France.Results: All of the 151 cases of hereditary haemoglobin disorders were categorized into four groups. Of those Haemoglobin-E ß thalassaemia was 64.90%, ß thalassaemia minor was 17.22%, Haemoglobin-E haemoglobinopathies was 11.92% and ß thalassaemia major was 5.96% of cases. Among 151 patients; 106 (70.20%) patients were severely anaemic, 30 (19.87%) patients were moderately anaemic and 15 (9.93%) patients were mildly anaemic. Among 98 patients of haemoglobin-E ß thalassaemia, 87 (88.76%) patients were severely anaemic.Conclusion: All anaemic patients especially who suffers from anaemia for a long period of time should be checked for haemoglobin disorders by routine haematological investigation and peripheral blood film examination. The patients with persistent anaemia and whose blood shows hypochromic microcytic anaemia should be advised for haemoglobin electrophoresis.KYAMC Journal Vol. 6, No.-1, Jul 2015, Page 545-549

Update on Malaria Diagnostics and Test Utilization.

Malaria is a potentially life-threatening disease requiring rapid diagnosis and treatment. Although microscopic examination of thick and thin blood films remains the gold standard for laboratory diagnosis, rapid antigen tests and nucleic acid amplification methods may also play a useful role in detection of acute infection. This review discusses the advantages and disadvantages of the commonly used diagnostic methods and provides important practice points for optimal malaria test utilization.

Autosomal recessive inherited bleeding disorders in Pakistan: a cross-sectional study from selected regions

BackgroundAutosomal recessive bleeding disorders (ARBDs) include deficiencies of clotting factors I, II, V, VII, X, XI, XIII, vitamin K dependent clotting factors, combined factor V & VIII, Von Willebrand Disease (vWD) type 3, Glanzmann’s thrombasthenia (GT) and Bernard–Soulier syndrome. Patients with primary bleeding disorders from all the major provincial capitals of Pakistan were screened for ARBDs. Prothrombin (PT), activated partial thromboplastin time (APTT), bleeding time (BT) and fibrinogen levels were measured. Cases with isolated prolonged APTT were tested for factors VIII and IX using factor assays This was followed by FXI:C level assessment in cases with normal FVIII and FIX levels. vWD was screened in patients with low FVIII levels. Factors II, V and X were tested in patients with simultaneous prolongation of PT and APTT. Peripheral blood film examination and platelet aggregation studies were performed to assess platelet disorders. Urea clot solubility testing was done to detect Factor XIII levels where platelet function tests were normal. Descriptive analysis was done using SPSS version 16.ResultsOf the 429 suspected bleeding disorder patients, 148 (35%) were diagnosed with hemophilia A and 211 (49.1%) patients had ARBDs. 70 patients (16.3%) remained undiagnosed. Out of 211 patients with ARBD; 95 (33.8%) had vWD type 3. Fibrinogen deficiency was found in 34 patients (12%), GT in 27 (9.6%), factor XIII deficiency in 13 (4.6%), factor VII deficiency in 12 (4.3%), factor V deficiency in 9 (3.2%). Eight patients (2.8%) had vitamin K-dependent clotting factor deficiency, Bernard–Soulier syndrome was diagnosed in seven patients (2.5%), factor X deficiency in 2 (0.7%), factor II deficiency in 2 (0.7%), factor XI deficiency and combined factor V and VIII deficiency in 1 (0.4%) patient each.ConclusionvWD type 3 was the most common ARBD found in our sample of patients in Pakistan, followed by fibrinogen deficiency and GT in respective order.

Oral route of transmission: Trypanosoma evansi in a mice model experiment.

Twelve Swiss albino mice of either sex and equal body weight were randomly divided in 2 groups (I and II), consisting of 9 and 3 mice respectively and were used to conduct the study. A dose of 2.5×104 number of Trypanosoma evansi was instantly fed to each mouse of group I. Each mouse of group II was inoculated intraperitoneally with same dose of parasites through infected mice blood and kept separate. The tail blood of each mouse was examined daily up to 30days post infection by examination of wet blood film and Giemsa-stained blood smears for presence of any trypanosomes. Out of 9 mice of group I those were infected orally, 3 (33.33%) mice became positive for presence of T. evansi both by examination of wet blood film and Giemsa-stained blood smears after 4, 6 and 7days post infection. After 2days post infection all intraperitoneally infected mice were found positive for T. evansi. Thus incubation period in orally infected mice was longer than the intraperitoneally infected mice. All the positive mice of both the groups died with high parasitaemia after 3-4days of first appearance of parasitaemia. From the present study, it can be concluded that besides mechanical or parenteral means of transmission, T. evansi could also be transmitted through oral route. Thus zoo carnivores might be infected with T. evansi and develop disease by eating infected blood or flesh of the infected animals, as a prey and predator relationship.

Prevalence of cryptosporidiasis and isosporiasis among HIV-positive patients attending some hospitals in Bauchi Metropolis

This study assess the effectiveness of insecticide-treated nets (ITNs) in reducing the prevalence of malaria among households following the Roll Back Malaria campaign on malaria prevention awareness and free distribution of ITNs in Ezza North L G A, Ebonyi State. Blood film examination was conducted on 715 persons and structured questionnaire administered to 304 household members. Data were analyzed using statistical package (SPSS) and presented as tables and significance of associations in parasitaemia prevalence. Results revealed that malaria parasite were observed in 175 (24.5%) out of 715 individuals examined. The highest prevalence of 8.3% was recorded among individuals aged 5-10 years. Females were more infected (12.9%) than their male counterpart (11.6%) with p=0.251. On ownership of ITNs, it was observed that 196 (64.5%) of the 304 households owned at least one ITNs and 152 (77.6%) households used their ITNs properly. Households who do not use bed net, stands 6.120 chance of been infected (ODD Ratio calculation). Malaria parasitaemia reported in this study is still high and sleeping under ITNs consistently has been found to significantly reduce the prevalence of malaria among the study-subjects. It is therefore recommended that ITNs campaign be intensified for increase in ITN ownership and usage among all individuals in the community.Keywords: ITN ownership; malaria prevalence; Ezza North; Ebonyi State.

Use of insecticide treated net (ITNS) among households and its impact on malaria prevalence in the communities in Ezza North Local Government Area of Ebonyi State, Nigeria

This study assess the effectiveness of insecticide-treated nets (ITNs) in reducing the prevalence of malaria among households following the Roll Back Malaria campaign on malaria prevention awareness and free distribution of ITNs in Ezza North L G A, Ebonyi State. Blood film examination was conducted on 715 persons and structured questionnaire administered to 304 household members. Data were analyzed using statistical package (SPSS) and presented as tables and significance of associations in parasitaemia prevalence. Results revealed that malaria parasite were observed in 175 (24.5%) out of 715 individuals examined. The highest prevalence of 8.3% was recorded among individuals aged 5-10 years. Females were more infected (12.9%) than their male counterpart (11.6%) with p=0.251. On ownership of ITNs, it was observed that 196 (64.5%) of the 304 households owned at least one ITNs and 152 (77.6%) households used their ITNs properly. Households who do not use bed net, stands 6.120 chance of been infected (ODD Ratio calculation). Malaria parasitaemia reported in this study is still high and sleeping under ITNs consistently has been found to significantly reduce the prevalence of malaria among the study-subjects. It is therefore recommended that ITNs campaign be intensified for increase in ITN ownership and usage among all individuals in the community.Keywords: ITN ownership; malaria prevalence; Ezza North; Ebonyi State.

Molecular Detection of Novel Genetic Variants Associated to Anaplasma ovis among Dromedary Camels in Iran.

To the best of our knowledge, little information is available regarding the presence of Anaplasma species in camels in Iran. This study sought to investigate the presence of Anaplasma species by microscopy and polymerase chain reaction (PCR) assays in 100 healthy dromedaries (Camelus dromedarius) arriving for slaughter. The microscopic examination of Giemsa-stained blood films revealed that Anaplasma like structures could be identified in the erythrocytes of two blood smears. To confirm the presence of and to identify the species of Anaplasma spp., a PCR technique was performed using primers amplifying a 750 bp fragment of the 16S rRNA gene of Anaplasma and the PCR products were analyzed by sequencing. The nucleotide sequence was compared to the sequences available in GenBank using the Basic Local Alignment Search Tool (BLAST). According to the results, the sequences of two 16S rRNA PCR products clearly fit within the Anaplasma genus in the family Anaplas mataceae. In this study, phylogenetic analysis using the 16S rRNA gene sequences revealed that two sequences obtained from monophyletic clusters included Anaplasma ovis (A. ovis). The obtained sequences had 99.6-100% similarity with previously published 16S rRNA gene sequences. This study aimed to evaluate the presence of novel genetic variants associated to A. ovis in dromedaries in the world. Further studies are recommended to establish the vector(s), as well as the veterinary and medical significance of these apparently novel variants in Iran.To the best of our knowledge, little information is available regarding the presence of Anaplasma species in camels in Iran. This study sought to investigate the presence of Anaplasma species by microscopy and polymerase chain reaction (PCR) assays in 100 healthy dromedaries (Camelus dromedarius) arriving for slaughter. The microscopic examination of Giemsa-stained blood films revealed that Anaplasma like structures could be identified in the erythrocytes of two blood smears. To confirm the presence of and to identify the species of Anaplasma spp., a PCR technique was performed using primers amplifying a 750 bp fragment of the 16S rRNA gene of Anaplasma and the PCR products were analyzed by sequencing. The nucleotide sequence was compared to the sequences available in GenBank using the Basic Local Alignment Search Tool (BLAST). According to the results, the sequences of two 16S rRNA PCR products clearly fit within the Anaplasma genus in the family Anaplas mataceae. In this study, phylogenetic analysis using the 16S rRNA gene sequences revealed that two sequences obtained from monophyletic clusters included Anaplasma ovis (A. ovis). The obtained sequences had 99.6-100% similarity with previously published 16S rRNA gene sequences. This study aimed to evaluate the presence of novel genetic variants associated to A. ovis in dromedaries in the world. Further studies are recommended to establish the vector(s), as well as the veterinary and medical significance of these apparently novel variants in Iran.

Comparative Use of RDT and Thick Film Microscopy in the Diagnosis of Malaria in Sub-urban Settlements in Makurdi, Nigeria

In as much as accurate diagnosis seems to be the only way of effecting rational therapy, it has been the most neglected area of malaria research. Thick blood film microscopy and Rapid diagnostic test (RDT) was comparatively used to study malaria prevalence in Makurdi, Nigeria. A total of 328 blood samples were collected from consented respondents and analyzed using Blood film examination with Field’s stains A and B staining techniques and Care Start TM rapid diagnostic test (RDT) manufactured by Access Bio Inc, USA to detect the presence of malaria parasites in blood. Questionnaires were used to get demographics of the respondents. Of the 328 participants examined, 164(50.0%) were positive for malaria parasites by light microscopy and 32(9.8%) were positive for malaria by RDT Care Start TM HRP2. The sensitivity and specificity of RDT was found to be 16.5% and 97.0% respectively while the Positive Predictive Value (PPV) and Negative Predictive Value (NPV) was found to be 84.4% and 53.7% respectively. The females 92(51.7%) and 20(11.2%) were slightly more infected than the males 72(48.0%) and 12(8.0%) using both methods, but result was not statistically significant (P>0.05). Malaria prevalence was higher among those with no formal education and least among those with tertiary education (P>0.05). The prevalence of malaria in respect to location was significantly higher in New GRA by RDT and Chile by light microscopy and least in High level and Agwan Jukum in order of RDT and microscopy (P=0.000). This study revealed that malaria can affect all sexes irrespective of their educational cadre and location. The Care Start TM RDT showed very poor sensitivity in contrast to light microscopy. In as much as light microscopy in poor set ups cannot be used routinely, the RDT has not proven to be a good replacement.

PARASITOLOGICAL AND MOLECULAR DETECTION OF THEILERIA EQUI IN DONKEYS IN ASSIUT GOVERNORATE

theileria equi is a tick-borne hemoprotozoan parasite and one of the causative agents of equine piroplasmosis. Data on T. equi infection in donkeys are scarce in Assiut governorate, Egypt. This study was conducted on the prevalence of T. equi infection in donkeys by using micropscopic examination and polymerase chain reaction (PCR) assay. Out of 50 donkeys, blood samples examined for T. equi, 14% (7/50) were positive by microscopic examination and 38% (19/50) by PCR. Prevalence of T. equi had high significant differences between microscopic examination and PCR assay. Prevalence of T. equi in male donkeys was 13.6% (3/22) by microscopic examination and 36.4% (8/22) by PCR. In female donkeys it was 14.3% (4/28) by microscopic examination and 39.3% (11/28) by PCR. No significant difference between the prevalence rate of infection in males and females were recorded. Microscopic examination of donkeys’ blood smears stained with Giemsa stain revealed forms of theilerial schizogony in lymphocytes (Kochs blue bodies) and intra-erythrocytic relatively small, less than 2 μm long pear-shaped and ring shaped merozoites of T. equi. The B1 gene specified for T. equi was detected by PCR in 19 blood samples. All positive samples with microscopic examination were also positive with PCR. The sensitivity, specificity and accuracy of PCR with respect to blood film examination were 61.3%, 100% and 82%, respectively. So, PCR was found to be more sensitive, specific and accurate than blood film examination.The present results suggest that T. equi parasite is widely spread in Assiut governorate due to high exposure to ticks.

Effect of malaria infection on hematological profiles of people living with human immunodeficiency virus in Gambella, southwest Ethiopia

BackgroundMalaria and human immunodeficiency virus are the two most devastating global health problems causing more than two million deaths each year. Hematological abnormalities such as anemia, thrombocytopenia and leucopenia are the common complications in malaria and HIV co-infected individuals. The aim of this study was to determine the effect of malaria infection on hematological profiles of people living with HIV attending Gambella Hospital ART clinic, Southwestern Ethiopia.ObjectiveTo determine the effect of malaria infection on hematological profiles of people living with HIV attending Gambella Hospital ART clinic, Southwestern Ethiopia.MethodsA facility based comparative cross-sectional study was conducted from May 25 to November 11, 2014 in Gambella Hospital. A total of 172 adult people living with HIV (86 malaria infected and 86 malaria non-infected) participants were included in the study. Demographic, anthropometric and clinical data were collected. Venous blood samples and stool specimen were collected for laboratory analysis. Microscopic examination of peripheral blood films was done for detection of malaria parasites. Descriptive statistics, student T- test, bivariable and multivariable analyses were performed using SPSS V-20. Statistical significance was set at p < 0.05.ResultsA total of 172 adult people living with HIV were included in the study. The prevalence of anemia, thrombocytopenia and leucopenia in malaria and HIV co-infected participants were 60.5%, 59.3%, and 43.0%, respectively. Resident (AOR: 4.67; 95% CI: 1.44, 15.14), malaria infection (AOR: 2.42; 95% CI: 1.16, 5.04) and CD4 + count were predictors for anemia. A predictor for thrombocytopenia was malaria infection (AOR: 9.79; 95% CI: 4.33, 22.17). Malaria parasitic density (AOR: 0.13; 95% CI: 0.03, 0.57) and CD4 + count (AOR: 4.77; 95% CI: 1.23, 18.45) were predictors of leucopenia.ConclusionsFindings suggest that the prevalence of anemia and thrombocytopenia were significantly higher in the malaria and HIV coinfected participants than the HIV mono-infected participants. Mean values of hematological profiles were significantly different in the two groups. Future prospective studies with larger sample size from other settings are needed to substantiate the findings.

A MALARIA VACCINE SITE CHARACTERISATION: PREVALENCE AND SPECIES DISTRIBUTION OF PLASMODIUM MALARIA IN A MALARIA ENDEMIC SETTING OF BURKINA FASO (WEST AFRICA)

BackgroundAny development of a vaccine strategy or its implementation is based on a good knowledge of the biology of the pathogen and its temporal and spatial distribution. In the case...

Seasonal fluctuation of trypanosomiasis in camels in North-West Egypt and effect of age, sex, location, health status and vector abundance on the prevalence

Follow-up surveys of the blood parasites of the livestock and their possible vectors are recommended to assess their distribution and infection rates in different areas in Egypt and to plan control measures against their vectors. By this study, the authors have completed a comprehensive survey of the extent of Trypanosoma evansi infection in North-West Egypt from November 2014 to October 2015, using blood film examination and RoTat1.2 PCR-based assay. The present study was carried out in the frame of PROCAMED project, supported by the European Union (ENPI-Joint operational Programme of the Mediterranean Basin-IEVP-CT). Historical data of our previous and the present survey have shown that trypanosomiasis in this particular area is constant and not subject to change over time. In addition, the present study has clarified that T. evansi is endemic in this area due to the interaction of a wide range of factors and features of the host which determine patterns of trypanosomiasis, and its distribution coincides with the distribution of vectors.

Utility of screening golf ball inclusions in hemoglobin H disease and its clinico-hematological profile

BACKGROUND: In India, hemoglobinopathy screening programs focus on reducing the burden of beta-thalassemia by screening the antenatal females and young adolescents. Although alpha (α)-thalassemia is common in the Mediterranean countries, alpha (α)-thalassemia screening and diagnosis are performed in a few centers in India. HbH disease is an uncommon clinical phenotype of α-thalassemia with variable clinical presentation. The hallmark feature of HbH is the presence of golf ball inclusions in the red cells demonstrated with supravital staining. PATIENTS AND METHODS: Here, we discuss an algorithmic approach to screen hemograms for thalassemic indices, followed by evaluation of reticulocyte preparation after 40 minutes (supravital stain preparation for golf ball inclusions). If golf ball inclusions were positive, Hb analysis on Hb-capillary zone electrophoresis (Hb-CZE)was done. RESULTS: We received 28,000 samples for complete hemogram and blood film examination, over a period of 8 months. Reticulocyte count was required in 11,600 samples out of which 13 cases were positive for golf ball inclusions. Of these, nine were identified as HbH disease on Hb-CZE. These patients presented with pallor and generalized weakness. On examination, jaundice (3/9 patients) and hepatosplenomegaly (4/9 patients) were relatively uncommon. The majority of them (6/9) required infrequent blood transfusions. All patients had microcytic hypochromic red cell indices with median HbA of 89% (range: 68.1%–96.9%), HbA2 of 1.4% (range: 0.8%–1.7%), and fast-moving band in zone 15 (HbH) of 5.6% (range: 1.4%–30.3%) on Hb-CZE. CONCLUSION: We emphasize a systematic screening of hemograms and peripheral smears along with a simple and cost-effective screening test for golf ball inclusions to diagnose HbH disease that often poses a clinical dilemma due to its variable clinical presentation.

Evaluation of Molecular Technique and Microscopical Examination for Diagnosis of Blood Parasites In Equine and Camels

This study was designed to evaluate PCR technique as a diagnostic tool for blood parasites infection in equine and camels in relation to conventional microscopic examination of stained blood smear. 150 equine stained blood smears were examined microscopically, 15 of them were suspected to be infested with equine piroplasmosis. 10 samples (including one positive by blood film) were examined by conventional PCR technique, 7 samples were positive. 135 camels stained blood smears were examined microscopically, 2 of them were infested with trypanosoma, and 10, 5 and 15 samples were suspected to be infested with theileria spp., babesia spp. and anaplasma spp., respectively. 10 samples (including 2 positive for trypanosoma), 10 negative for theileria, 10 negative for babesia and 10 negative for anaplasma by blood film were examined separately by conventional PCR technique, 9, 1, 1 and 5 samples were positive, respectively. The results concluded that PCR assays are more sensitive than blood film examination.