Giemsa Research Articles

Bovine cerebral theileriosis: first molecular report in cross bred cattle calf in India.

Bovine tropical theileriosis caused by Theileria annulata, is a serious constraint to Indian dairy industry with more fatal infections in exotic cattle and substantial losses to cross-bred and indigenous zebu cattle. The present communication is to place on record the first report of molecular based confirmed case of cerebral theileriosis caused by T. annulata coupled with its morphological detection, clinical manifestations, haematological alterations and therapeutic management in a cross bred cattle calf from India. After preparation of peripheral thin blood smear from cross bred cattle calf at the site of collection and fixation with methanol, blood sample brought to Department of Veterinary Parasitology, College of Veterinary Science and A.H, Jabalpur and stained by standard protocol for Giemsa staining. Genomic DNA was isolated from the collected blood sample using QIAamp® DNA blood mini kit following the manufacturer's recommendations and PCR was performed. The cross bred cow calf revealed high rise in temperature (105.5°F), increased heart rate, labored breathing with seromucous nasal discharge, enlargement of prescapular lymph node and animal exhibited tonic clonic convulsions in response to any sudden noise. Giemsa stained thin blood smear revealed intraerythrocytic piroplasm and Koch'sblue bodies of T. annulata within the cytoplasm of lymphocytes. The species of Theileria was confirmed by molecular amplification of genomic DNA as T. annulata.

Morphological and Morphometric Analysis of Trypanosoma lewisi and Toxoplasma gondii in Malang City, Indonesia Rats

Rats are reported to be the intermediate hosts and reservoirs of several zoonotic protozoal diseases. Trypanosoma lewisi is commonly reported in rats’ blood and is considered non-pathogenic protozoa in humans. However, some countries documented several cases in humans with T. lewisi infection. Another zoonotic protozoon that develops in rats and can be transmissible to humans is Toxoplasma gondii. We intended to present the morphology and morphometry of T. lewisi and T.gondii in wild rats collected around Malang City to explore the potential risk of transmission nearby. The rats were collected using single live traps followed by identification, sexing, age approximation, and body morphometry. All specimens were euthanized according to the standard procedure followed by blood and peritoneal fluid collection. The fluid smear preparation and Giemsa staining were performed to detect the presence of T. lewisi and T. gondii. Morphologic and morphometric analyses were conducted using ImageJ software. Among the collected 50 collected rats, 23 were identified as Rattus norvegicus (46%), 22 as Rattus rattus (44 %), and 5 as Mus musculus (1%). In the case of protozoans infection, ten individuals were infected with T. Lewisi (20%) from the blood smear check, whereas peritoneal fluid smear examination revealed an infection of T. gondii in a specimen (2%). Results of the study proved trypanosomiasis and toxoplasmosis in wild rats in Malang City. Though the clinical significance to human and public health impact is questionable, further research and surveillance of rodent-borne parasitic pathogens will provide more information for pre-emptive action.

Differences in mouse strains determine the outcome of Der p 2 allergy induction protocols

In vivo animal models can provide worthy information on various aspects of asthma mechanism and pathogenesis. The genetic predisposition and phenotype of mice may affect the immune response itself. Here we compare the early immune response to Der p 2 or HDM allergen extract upon injection and inhalation in BALB/c and C57BL/6 mice. Female C57BL/6 and BALB/c mice were immunized with Der p 2 allergen subcutaneously followed by inhalation of Der p 2 or HDM extract. After challenge, the mice were euthanized; blood, bronchoalveolar lavage (BAL), spleens and lungs were collected. Cells from BAL were identified by May-Grünwald Giemsa staining and lung leukocyte populations were analyzed by flow cytometry. Serum antibody levels of Der p 2 specific IgE, IgG, IgG1 and IgG2a were assessed by ELISA, and cytokine secretion (IL-4, IFN-γ and IL-10) was evaluated upon stimulation with Der p 2 or HDM extract. The Th2 immune response was confirmed by elevated allergen-specific immunoglobulin E (IgE) and the allergic reaction was evidenced by infiltration of eosinophils and/or neutrophils into BAL. We found that BALB/c mice were inefficient in integrating local with systemic immune response, evidenced by almost no IgG or IgE production upon one subcutaneous injection and subsequent inhalation of Der p 2 allergen; also, the bronchoalveolar lavage infiltrate in these mice consisted of neutrophil infiltration, unlike C57BL/6 mice in which eosinophilic infiltrate predominated. The differences between BALB/c and C57BL/6 mice strains could be exploited for generating different types of responses to the Der p 2 allergen.

Histological stains and their application in teaching and research

This review aimed to highlight the historical perspectives focused on histological stains and their application in teaching and research. While early histologists used indigenous substances like alizarin, carmine, and saffron to stain tissues, modern histologists utilize techniques that study cellular organelles and substances within tissues. Despite the advent of new staining techniques, the modification of old ones, and decrease in the use of natural dyes in the 20th century, some of the stains used in the past are still very much in use. These techniques are used in teaching and research for the demonstration and display of the different substances within tissues. These include the Giemsa stain for parasites and fungi, the Trichrome stain for collagen, oil red O stain for lipids, periodic acid Schiff for glycogen, Crystal violet for Nissl granules, Golgi stain for neuronal cells, as well as routine hematoxylin, and eosin (H&E) for a general histology. Histological stains can be used to teach histology, cellular pathology, microbiology, and parasitology. They can also be used to study the rate of progression of disease or the healing process of diseased conditions.

Prevalence Of Malaria And Intestinal Parasitic Co-Infection Among Diabetic Patients In Calabar.

Background: Malaria and intestinal parasitosis is a public health problem among diabetic patients, therefore, this work evaluates the prevalence of intestinal parasitic infection and malaria co-infections in diabetics.
 Materials and Methods
 Capillary blood and fecal samples were collected from 190 diabetic patients at the outpatient clinic of University of Calabar Teaching Hospital and Navy Reference Hospital Calabar and another batch of capillary blood and fecal samples from 110 non-diabetic subjects. The stool samples were examined macroscopically and microscopically by direct smear and formol- ether concentration method and also stained by modified Ziehl- Neelsen acid fast stain. The thick and thin blood film were stained with 10% Giemsa stain and viewed microscopically.
 Result: Amongst the test subjects, malaria parasites had a prevalence rate of 30 (15.8%), intestinal parasites had a prevalence rate of 48 (25.3%), and prevalence of co-infection with malaria parasites and intestinal parasites were 8 (4.2%), the difference was statistically significant (p= 0.036). Amongst the non-diabetic subjects, prevalence of malaria parasites was 12 (10.9%), intestinal parasites had a prevalent rate of 12 (10.9%) and a co-infection prevalence rate of 12 (3.6%). Amongst the diabetic patients, subjects aged 21-30 years had the highest infection rate of 14 (70.0%) for malaria parasites while 31-40 years had the highest infection of 8 (57.1%) for intestinal parasites the difference was statistically significant p = 0.0001. Amongst the non-diabetic subjects, age group 31-40 had the highest prevalence rate for malaria parasites 4(28.6%) and intestinal parasites 6(42.9%), while age group 31-40 and 41-50 had the highest prevalent rates of 2(14.3%) for co-infection. Amongst the diabetic patients, male subjects had a higher prevalence rates for malaria parasites, intestinal parasites and co-infection of 20(33.3%), 26(43.3%) and 6(10.0%) respectively which was statistically significant (p= 0.001). For the non-diabetic subjects, females had a higher prevalence rate for malaria infection 8(13.3%), while males had a higher prevalence rate for intestinal parasites and co-infection of 8(16.0%) and 4(8.0%) respectively, the difference was not statistically significant (p=0.250). Hookworm was the most observed parasite, 18(37.5%), Ascaris lumbricoides, 14 (29.2%), Cryptosporidium parvum, 8 (16.7%), Cyclospora cayetanensis, 4 (8.3%), Trichuris trichiura, 2 (4.2%) and Microsporidia, 2 (4.2%).
 Conclusion: Based on findings in this study, this work has shown a prevalence of intestinal parasites (25.3%), and prevalence of malaria infection (15.8%) in diabetic subjects; and this study also illustrate the prevalence of malaria and intestinal parasitic co- infection of 4.2%, and the need to put in place strategies for the control of the parasite among this group of patients.

Malaria and anemia in children under 7 years of age in the western region of Côte d’Ivoire

BackgroundAnemia is a major public health problem, affecting nearly one-quarter of the world’s population. It is defined as a reduction in the hemoglobin level in the peripheral blood to below the normal threshold set for a particular population. Very often in the subtropics, helminths or malaria co-infect an individual, causing morbidities that vary by age and region. This study aims to characterize the type of anemia observed in children under 7 years of age infected with malaria in the western region of Côte d’Ivoire, to recommend a better strategy of care.MethodsThe study was carried out from March 2020 to May 2021 in 22 villages in Man, Tonkpi Region, with a cohort of 451 children, both male and female, aged from 3 months to 6 years. The children provided venous blood samples for the diagnosis and characterization of anemia (full blood count), and Giemsa staining (GS) (thick and thin smears) and rapid diagnostic tests (RDTs) were used for the diagnosis of malaria. Risk factors and morbidity profiles were assessed using a questionnaire. Logistic regressions models were employed to identify independent risk factors and morbidity patterns associated with Plasmodium falciparum mono-infection and co-infections.ResultsOf the 451 children who completed the study, 221 (49.0%) were female and 230 (51.0%) were male. The prevalence of anemia was 55.0%, distributed as 30.7% mild, 66.1% moderate, and 3.2% severe anemia. The characterization of anemia revealed that hypochromic microcytic anemia (HMA) was the predominant type, being found in 195 (78.63%) children. It was followed by normochromic microcytic anemia (29 children, 11.69%), normochromic normocytic anemia (14 children, 5.65%), and, finally, hypochromic normocytic anemia (10 children, 4.03%). The prevalence of malaria was 66.7% and 78.3% based on GS and RDTs, respectively. The closed association between malaria (Plasmodium) and anemia led to P. falciparum alone causing 56.7% of mild, 51.3% of moderate, and 37.5% of severe anemia in children.ConclusionMalaria infection was highly prevalent among children aged ≤ 7 years in both sex and in different age groups, although the number of Plasmodium parasites present during infections was greatest in younger children. Similarly, the prevalence of anemia was high, with moderate anemia and HMA being more prevalent in children ≤ 7 years of age in the western region of Côte d’Ivoire.

The Effects of Saposhnikovia divaricata Aqueous Extracts on the Inflammation and Intestinal Microflora in Allergic Rhinitis Mice.

Background Allergic rhinitis (AR) is a type I allergic disease induced by IgE. Traditional Chinese medicine Saposhnikovia divaricata (Turcz.) Schischk (SD) has anti-inflammatory and antiallergic effects. Materials and Methods AR model mice were constructed with ovalbumin (OVA) sensitization to observe the improving effect of SD treatment on AR by counting the number of sneezing and rubbing the nose, hematoxylin-eosin, periodic acid-Schiff, and toluidine blue stains. In addition, the allergy and inflammatory cytokines levels and inflammatory cells were observed by ELISA and Wright's-Giemsa stain. The protein levels of the TLR4/TRAF6/NF-κB and IL-6/ROR-γt/STAT3 pathways were measured by immunohistochemistry, quantitative real-time PCR, and western blot. The intestinal microflora abundance in mice was observed by 16S rDNA high-throughput sequencing. Results SD treatment inhibited the sneezing and rubbing times of the nose, decreased the degree of a dense arrangement of cells and mucosal swelling and the number of goblet and mast cells of nasal lavage fluid, reduced the levels of IgE, histamine, Leukotriene B4, IL-4, IL-5, TNF-α, IL-6, and IL-17, the eosinophils, neutrophils, and lymphocytes number, the LR4, TRAF6, IL-6, ROR-γt, and STAT3 mRNA levels, respectively, while, it increased the IL-2, IL-10, IFN-γ, and TGF-β1 proteins. SD treatment inhibited the NF-κB, p-STAT3, TLR4, TRAF6, and p-IκBα/IκBα proteins. Besides, the effects of OVA and SD treatments were significantly correlated with the abundance of intestinal microflora. The abundances of Cytophagales, Burkholderia, Alteromonadales, Lactococcus, and Clostridiaceae were changed in SD treatment on AR mice. Conclusions This study provides a possibility that the improvement effect of SD treatment on allergies and inflammation in AR mice may be related to the TLR4/TRAF6/NF-κB and IL-6/ROR-γt/STAT3 pathways and intestinal microflora modulation.

Insulin-like growth factor 1 serum levels in different stages of gastric cancer and their association with Helicobacter pylori status

High serum insulin-like growth factor 1 (IGF-1) and positive Helicobacter pylori (H. pylori) may increase the risk of gastric cancer (GC). We aimed to investigate IGF-1 serum levels in different stages of GC patients and their association with H. pylori status. A total of 90 participants, including 60 GC patients and 30 noncancerous (NC) individuals, were included in the present study. IGF-1 serum levels and candidate proteins were assessed using enzyme-linked immunosorbent and immunohistochemistry techniques. Likewise, Giemsa staining was applied to detect H. pylori infection. The candidate genes' expression, including IGF-1R, PI3KCA, AKT1, mTOR1, KRAS, BRAF, and ERK1, was also evaluated by a real-time PCR assay. The results of advanced GC stages indicated a significantly high IHC score for IGF-1R and phosphorylated AKT, mTOR, and ERK proteins compared to the early stages. Moreover, IGF-1 serum levels and the expression of candidate genes were considerably increased in the advanced GC patients compared to the early stages and the positive H. pylori status compared to the negative H. pylori status (P < 0.05). As a result, high IGF-1 serum levels and positive H. pylori status may be correlated with gastric tumor progression, and the inhibition of IGF-1 and the eradication of H. pylori infection might be new therapeutic targets in GC patients.

Fungal Elements in Sputum Cytology Among Active and Post_Treated Pulmonary Tuberculosis Patients in Al Managil Teaching Hospital, Gezira State, Sudan (2020)

Sputum cytology is still one of the most effective and producible sample for diagnosis and evaluation of lung diseases and disorders. For this, it can used for evaluation of apportunistic fungal pathogens on pulmonary tuberculosis patients, which is of leading cause of death worldwide. On this cross sectional laboratory base study, which used to evaluate the sputum cytology for presence of opportunistic fungal elements. A total 110 early morning expectorate sample collected from the period 1/3 to 30/6/2020. 69/110 (63%) from patients come for first diagnosis to the center of T.B and HIV. Al Managil teaching hospital, and 41/110 (37%) follow-up starting from second month until six month post- treated follow up. From each sample two slides was prepared. One of them fixed immediately before air-drying in 95% ethanol and later stain by PAS technique, the other fixed after air-drying on absolute methanol and stain by Giemsa stain. Regardless to other method of fungal identification such as serology or mycological culture, only depends on microscopic identification. The study found that 95/110 (86%) was negative for fungal elements, and 15(14%) was positive, 9(8%) positive in diagnosis group and 6(5%) in follow-up group. Moreover the most common infectious agents was Candida species 9/110 (8%), 6/110 (5%) present as yeast and 3/110 (2%) as Pseudohyhae. Followed by Aspergillus species 5/110 (4%) then actinomyces species 1/110(.9%). The incidence of infection is higher in rural (82%) males (55%) farmers (36%). In the majority of the pateints with negative results for fungi MDR-TB not detected 87(79%), followed by positive for fungi and also MDR-TB not detected 15 (13.6%), then (negative and positive) for fungi and MDR-TB with high rate with the same percentage 3(2.8%), and finely very low MDR-TB 1(.9%), medium MDR-TB 1(.9%) without any detection of fungal elements. The study recommended including fungal testing and antifungal drugs on the pulmonary TB treatment plan as possible causes of complications.

An In Vitro Study on Sonodynamic Therapy of Leishmania tropica Using Curcumin

Leishmaniasis is an infectious disease that is transmitted by Phlebotomus, 400 thousand new cases appearing every year, and approximately 350 million people are at risk, and accepted by the World Health Organization as one of the six important tropical diseases. Cutaneous leishmaniasis is a disease that occurs on exposed areas of the body and is characterized by long-term non-healing skin lesions. Although the treatment methods applied today vary according to the clinical picture of the patient, the immune system of the person and the causative agent Leishmania species, there is still no standard treatment scheme that has few side effects and can be used in the treatment of leishmaniasis. Therefore, alternative treatment methods with less side effects are being tried. Sonodynamic therapy (SDT) has also emerged as an active antimicrobial research area in recent years. SDT, a new modality for antibacterial therapy, aims to increase antibacterial effects with the simultaneous combination of low-intensity ultrasound and sonosensitizer. There is no information in the literature about the effect of SDT on parasites. In this study, it was aimed to demontrate the anti-leishmanial effect and possible mechanisms of curcumin mediated SDT on L.tropica promastigotes in vitro. Parasites were incubated with 0.25, 1.0, 4.0 and 15.6 micromolar (μM) of curcumin for one hour and subjected to 1 MHz frequency, 50% duty cycle and 3 W/cm2 intensity ultrasound irradiation. XTT assay was used to evaluate the viability of the cells and morphological changes were analyzed by Giemsa staining. Flow cytometry was used to quantify the fluorescence emitted by intracellular reactive oxygen species (ROS) signal, JC-1, cell cycle, Annexin V/PI staining reagents. With the combination of curcumin (15.6 μM) and ultrasound (3 W/cm2 intensity, seven minutes), L.tropica promastigote viability was found to be significantly decreased compared to the control group. Giemsa staining results showed that 15.6 μM curcumin mediated SDT induced several morphological alterations in L.tropica promastigotes typical for apoptosis. Late apoptosis was observed in 15.6 μM curcumin combined SDT treated parasites according to Annexin/PI staining. Besides, curcumin mediated SDT caused mitochondrial membrane potential (∆ᴪm) loss. Cell cycle analysis data indicated that curcumin based SDT caused an subG1 arrest in the cell cycle of L.tropica promastigotes. The generation of intracellular ROS detected by flow cytometry was increased in L.tropica promastigotes treated with curcumin mediated SDT. This study provided new data elucidating the molecular mechanism underlying the anti-leishmanial effect of curcumin mediated SDT. Curcumin mediated SDT has the potential to inactivate L.tropica promastigotes. However, further testing with amastigote or animal models is needed.

Phenotypic Characterization and Isolation of Myeloid-Derived Suppressor Cells.

Myeloid-derived suppressor cells (MDSCs) are heterogenous populations of immature myeloid cells that can be divided into two main subpopulations, polymorphonuclear (PMN) MDSCs and monocytic (M) MDSCs. These cells accumulate during chronic inflammation and induce immunosuppression evident in an array of pathologies such as cancer, inflammatory bowel disease, and infectious and autoimmune diseases. Herein, we describe methods to isolate and characterize MDSCs from various murine tissue, as well as to phenotype blood-derived MDSCs from patients. The protocols describe methods for isolation of total MDSCs and their subpopulations, for characterization, and for evaluation of their distribution within tissue, as well as for assessing their maturation stage by flow cytometry, immunofluorescence analyses, and Giemsa staining. © 2022 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Single-cell suspension generation from different tissue Alternate Protocol 1: Single-cell suspension generation from subcutaneous melanoma tumors Basic Protocol 2: Characterization of MDSC phenotype Basic Protocol 3: Cell separation using magnetic beads: Separating pan-MDSCs or PMN-MDSC and M-MDSC subpopulations Alternate Protocol 2: Staining and preparing MDSCs for sorting Support Protocol: PMN-MDSC and M-MDSC gating strategy in mouse Basic Protocol 4: Immunofluorescence analysis of MDSCs Basic Protocol 5: Handling human blood samples and characterizing human MDSCs Alternate Protocol 3: Flow cytometry staining of thawed human whole blood samples.

S2759 Case of Concomitant Cryptosporidiosis and Ulcerative Colitis

Introduction: Cryptosporidiosis is a parasitic infection of gastrointestinal epithelia that presents primarily as acute watery diarrhea. In patients with inflammatory bowel disease (IBD), cryptosporidiosis can be mistaken for a flare. We present a case of an active duty service member with positive Cryptosporidium PCR on workup of what appeared to be an ulcerative colitis (UC) flare. Case Description/Methods: A forty-three year-old male presented after two weeks of acutely worsening diarrhea, rectal pressure, blood with wiping, and bloating. Testing for Cryptosporidium was negative by routine histology and Giemsa and GMS stains of duodenal biopsies but was positive on a stool PCR panel. Both calprotectin and CRP were elevated. Colonoscopy showed inflammation to 30cm with friability, spontaneous hemorrhage, and ulceration. He was started on nitazoxanide for Cryptosporidium and oral mesalamine and a prednisone taper for UC flare. He improved symptomatically, with less frequent cramping and bowel movements and more formed stools. Discussion: This case resembles documented cases of cryptosporidiosis in IBD patients but is unique in that most other cases were in Crohn’s patients 2-5,7-9. Our patient’s improvement on the three medications makes it difficult to definitively attribute the patient’s symptoms to either cryptosporidiosis or UC flare. Combining nitazoxanide with immunosuppression has not worsened his disease severity, which is also reflected in literature 9. It is possible, however, that nitazoxanide alone would have resolved his symptoms and obviated steroid use 3. The decision whether to test for Cryptosporidium in suspected UC flare should be based on pre-test probability and type of testing available. As seen in our case, less sensitive tests like Giemsa and GMS staining may result in false negatives. Stool PCR is the gold standard for detecting Cryptosporidum. PCR does not depend on fecal oocyst shedding, unlike modified Ziehl-Neelsen acid fast stain, direct immunofluorescence antibody testing, and stool microscopy 1,6. The weakness of stool PCR is that it also detects nonviable Cryptosporidium, and a positive test does not necessarily confirm an infectious cause of diarrhea. Lessons from this case include the importance of correct diagnosis to guide treatment, consideration of Cryptosporidium testing in cases of suspected UC flare, and the range of sensitivities among Cryptosporidium tests.

The Isolation and Preparation of Samwinol from Dracocephalum heterophyllum and Prevention on Aβ25-35-Induced Neuroinflammation in PC-12 Cells.

Dracocephalum heterophyllum (D. heterophyllum) is a traditional Chinese Tibetan medicine that has been used for the treatment of lymphitis, hepatitis, and bronchitis. However, only a few selected chemical components are currently obtained from D. heterophyllum, which limits its further pharmacological applications. In this study, we have obtained samwinol from D. heterophyllum by medium- and high-pressure liquid chromatography separation for the first time. Thereafter, we investigated the protective actions of samwinol against amyloid beta protein fragment 25–35 (Aβ25–35) induced neurotoxicity in cultured rat pheochromocytoma PC-12 cells and explored its underlying mechanisms of action. The results indicated that samwinol could increase cell viability and inhibit the production of reactive oxygen species (ROS) and mitochondria-derived ROS, as assessed by MTT assay, Giemsa staining, and flow cytometry assay. Through Western blot analysis, it was found that samwinol substantially inhibited the phosphorylation of ERK(1/2) and promoted the expression of HO-1 and Nrf2. The data obtained from molecular docking were also consistent with the above conclusions. All of these results showed that samwinol from D. heterophyllum can display significant anti-neuroinflammatory and antioxidant activities in vitro, which are associated with the suppression of ERK/AKT phosphorylation and the activation of the Nrf2/HO-1 signaling pathway. In the future, additional in-depth mechanism studies will be carried out to provide more evidence for the potential of samwinol in the treatment of Alzheimer’s disease.

Prevalence and Risk Factors of Malaria among Pregnant Women receiving Antenatal Care in a Health Facility in Delta State, Southern Nigeria

Background: Malaria is one of the major causes of morbidity and mortality in developing countries including Nigeria. Malaria in pregnancy is estimated to cause about 15% of maternal deaths globally, accounting for over 10,000 maternal and 200,000 neonatal deaths annually. This study determined the prevalence and risk factors of malaria infection among pregnant women receiving antenatal care in a health facility in Delta State. Methods: This was a facility-based cross-sectional study carried out among 418 pregnant women who were selected by systematic sampling technique. Clinical malaria was confirmed using microscopy method (Giemsa staining technique) while a pre-tested, structured, interviewer administered questionnaire was used to collect information on soscio-demographic characteristics and obstetrics history of the respondents. Data were analyzed using Statistical Package for the Social Sciences (SPSS) version 20.0 and the level of statistical significance was set at p&lt; 0.05. Results: The mean age of the participants was 29.9 ± 5.7 years. Two hundred and sixty three (62.9%) pregnant women tested positive for malaria. Respondents who were pregnant for the second time (AOR = 0.521, 95% CI: 0.28 – 0.99, p = 0.045) and those from a family size of 1-6 (AOR = 2.123, 95% CI: 1.12 – 4.04, p = 0.022) were more likely to test positive to malaria parasite. Conclusion: In this study, the prevalence of malaria in pregnant women was high and the significant predictors were; family size of 1–6, skill level 1, first trimester and multigravidae. The Delta State Ministry of Health should ensure increasing access to IPT, ITNs/LLIN and health education on malaria in all hospitals rendering ANC services.

Immunomodulatory Potency Etlingera rubroloba A.D. Poulsen Fruit Ethanol extract against Macrophage Phagocytic Activity and CD4 Levels in Wistar Male Rats

Etlingera rubroloba A.D. Poulsen is an endemic plant in Southeast Sulawesi and taxonomic affinity with Etlingera elatior (Jack) R. M. Smith. The fruit of E. elatior has been reported to provide immunomodulatory activity by increasing the phagocytic activity of macrophage cells. This study aimed to determine the immunomodulatory potency of the E. rubroloba fruit ethanol extract with parameters of phagocytic activity and CD4 levels. This study used 24 male Wistar rats divided into 6 treatment groups, namely normal, negative (Na-CMC 0.5%), positive (Phylantii extract 0.135 mg/kgbw), extract dose I (200), dose II (300), and dose III (400) mg/kgBW. The treatment was given 3 mL each orally for seven days. All mice (except the normal group) were infected with 0.5 mL of Staphylococcus aureus intraperitoneally on the eighth day. The data were statistically analyzed by measuring macrophage phagocytic activity microscopically using Giemsa staining and CD4 levels using the sandwich ELISA method. The results showed that dose 300 mg/KgBW demonstrated the highest average percentage of macrophage cell phagocytic activity and CD4 level, which were 80.5% and 227.49 ng/mL, respectively. They were higher than the positive control treatment. Based on the results of Tukey's post hoc statistical test, all treatment groups had significant differences from the positive control group in both parameters (P&lt;0.05). This study concludes that the ethanolic extract of the fruit of E. rubroloba dose of 300 mg/kgbw has potential as an immunomodulator where increased phagocytic activity is associated with increased CD4 levels.

Point prevalence mapping of malaria infection in Rivers State, Nigeria

Background: Variations in the risk of malaria across locations exist but are poorly understood though identifying hotspots of malaria transmission will create opportunities for targeted interventions. Point prevalence of malaria in Rivers State was studied using Primary Healthcare Centres (PHCs) as survey points.&#x0D; Methods: The PHCs in Rivers State were geo-referenced using a handheld Global Positioning System (GPS) and 74 were selected across 21 local government areas using systematic grid point sampling. Blood samples were obtained from 2340 persons who consented and questionnaires were administered to obtain their demographic data. Malaria parasites in blood films were detected using the Giemsa staining technique. Data generated were analysed using SPSS 22.0 and presented using descriptive statistics. The level of relationship amongst the parameters was obtained using Chi-square. Co-ordinates of PHCs sampled and their prevalence data for malaria were entered into Microsoft Excel 2007 spreadsheet and transmitted to ArcGIS 10.8. This platform was then used to produce point prevalence infection maps of the State using geographic information systems (GIS). Survey points with malaria point prevalence values of 75% and above and cumulative prevalence of 1.97% and above were categorised as malaria transmission hot spots in the various LGAs.&#x0D; Results: The study recorded an overall prevalence of 56.3%, with P.falciparum as the only identified malaria parasite. Data revealed that Oyoro Model Primary Health Centre (MPHC), Arukwo Primary Health Centre, Ele Health Post (HP) and Emago HP recorded very high prevalence of 96.7%, 96%, 95.2% and 94.4% respectively, whereas MPHC Iriebe had the least prevalence. Twelve hotspots with point prevalence above 75% were identified and eight hotspots likewise with cumulative prevalence above 1.97%.&#x0D; Conclusion: Malaria infection remains endemic in Rivers State. This study provides malaria point prevalence maps of Rivers State which will serve as a reference to policymakers for strategic interventions in the State

First Report on Classical and Molecular Cytogenetics of Doi Inthanon Bent-toed Gecko, Cyrtodactylus inthanon Kunya et al., 2015 (Squamata: Gekkonidae) in Thailand

This study analyzed the karyotype of Cyrtodactylus inthanon Kunya et al., 2015 from Doi Inthanon, Chiang Mai Province, northern Thailand. The metaphase and meiotic chromosome preparations were obtained by squash technique from bone marrow and testes, respectively. The chromosomes were stained by Giemsa staining, Ag-NOR banding and molecular cytogenetics with ?uorescence in situ hybridization (FISH) using microsatellites d(CA)15, d(GC)15, d(CAG)10 and d(GAA)10 as probes. The results showed the diploid chromosome number (2n) of 40. The chromosome types of metacentric, submetacentric, acrocentric and telocentric chromosomes were 12-4-2-22, respectively. The Ag-NORs banding technique provides the pair of nucleolar organizer regions (NORs) on the telomeric region of the long arm of acrocentric pair 12. There are no sex differences in karyotypes between males and females. We also found that during metaphase I on meiosis of C. inthanon, the homologous chromosomes appeared synapsis of 20 bivalents. The microsatellite d(CA)15 signals were located on the sub-centromeric region of the metacentric pair 10, whereas the d(GC)15, d(CAG)10 and d(GAA)10 repeats are highly accumulated throughout almost all entire chromosomes. The karyotype formula is as follows: C. inthanon (2n = 40), Lm2 + Lsm4 + Lt6 + Mm2 + Mt8 + Sm8 + Sa2 + St8.

P186 Disseminated Histoplasmosis in an immunocompetent patient in a tertiary care center in North India

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM ObjectivesHistoplasmosis is a geographically restricted dimorphic fungi that causes disseminated infection in immune-competent as well as immunocompromised patients. Adrenal involvement is seen in disseminated disease but sometimes it may be the only site where then infection can be demonstrated. Early diagnosis and treatment are needed to save the patient from fatal adrenal insufficiency. We present a case of bilateral adrenal histoplasmosis in immunocompetent patient.MethodsA 63-year-old male presented to our hospital with a history of insidious onset of decreased appetite and unintentional weight loss for the last 6 months associated with generalized weakness. Patient had a history of mild to moderate intensity epigastric pain and discomfort which was intermittent in nature. Patient originally belongs to Azamgarh, Uttar Pradesh, but he was residing in Kolkata for the last 8 months.Patient had no history of fever, cough, hemoptysis, jaundice, chronic diarrhea, and steatorrhea. No history of orthostatic hypotension, salt craving, hyperpigmentation, headache, visual field disturbances, polyuria, behavioral changes, episodic headache, palpitation, diaphoresis, systemic hypertension with episodic all four-limb weakness. There was no history of abdominal striae, easy bruisability, difficulty in standing from squatting position. There was no history of tuberculosis among family members. On detailed history, it was revealed that he fed pigeons every day in the slum house where he lived in Kolkata.On the CECT abdomen it was found that there is an ill-defined hypodense enhancing lesion (72 × 52 × 77 mm) in right suprarenal region and bulky, necrosed 25 × 26 × 19 mm lesion in left suprarenal gland associated with multiple nonnecrotic paraaortic and aortic caval lymphadenopathy. He also received empirical anti-tubercular therapy for 15 days in the form of ethambutol and levofloxacin.ResultsIn all, 10% KOH wet mount of crushed smear of adrenal biopsy samples showed tissue debris and small narrow neck budding yeasts. Giemsa stain shows few small budding yeasts. Culture was put in SDA at 25°C and 37°C and incubated. On day 12, growth of colony in 25°C appears as white cottony growth with yellowish white reverse. On day 24, colony appears as buff brown with yellowish brown reverse. LPCB was done from the colony showing presence of characteristic tuberculate macroconidia (8–14) μm in diameter formed on short, hyaline, undifferentiated conidiophores and production of plenty round to pyriform microconidia (2–4 μm) in diameter, occurring on short branches and directly on the sides of the hyphae. Based on the direct microscopy and culture characteristics a diagnosis of Histoplasma capsulatum was given.ConclusionSystemic histoplasmosis is typically acquired through inhalation of microconidia or small hyphal elements in soil contaminated with bird and bat droppings leading to primary infection. This patient only manifested bilateral adrenal involvement with nonspecific symptoms.

Comparison of Diagnostic Methods for Detection of Trichomonas vaginalis in Prediagnosed Vaginitis Cases and Its Association with Various Pathogens

Parasitological diagnostic methods such as direct microscopy, staining examination and culturemethodsare frequently used in the diagnosis ofTrichomonasvaginalis(T. vaginalis).Though, nowadays, new diagnostic methods, especially DNA-based methods, are developing, enabling the simultaneous recognition of different pathogens.In our study,weevaluatedwhetherthe choice of multiplex polymerase chain reaction (PCR), in whichT. vaginalisand different pathogens can be detected,is be an alternativeto classical methods and to evaluate the possible coexistence of pathogens. In our study, swab samples taken during routine examination of 100 female patients who presented to Manisa Celal Bayar University and Manisa City Hospital Outpatient Clinics Obstetrics and Gynecology were evaluated.The presence ofT. vaginaliswas investigated in these samples by direct microscopy, Giemsa stain and culture.BesidesT. vaginalis, other possible agents were also investigated by real-time multiplex PCR method. At least one agent was detected in 85 (85%) of the 100 patient samples included in our study.T. vaginalis positivity was detected in 6 (6%)of thesamples by parasitological diagnosis methods and in 10 (10%)of thesamples by multiplex PCR.Additionally,with real-time multiplex PCR,Chlamydia trachomatisin 4 (4%),Neisseria gonorrhoeaein 3 (3%),Ureaplasma urealyticum/parvumin 68 (68%),Gardnerella vaginalisin 68 (68%) andHerpes simplex virus1/2 in 1 (1%) of thesamplepositivity was found.Mycoplasma genitalium, another agent examinedbymultiplex PCR, was not found positive inany sample.The Kappa value of theculture thatis a parasitological test and multiplex PCR forT. vaginalisshowed moderate agreement with 59.5%. It has been concluded that usingreal-time multiplex PCR method, which hashigh specificity and sensitivity, in addition to microscopy and culture methods in the diagnosis ofT. vaginalis, could contribute to the correct and effective treatment by detecting multiple infections.

Urine cytological study in patients with clinicopathologically confirmed neuronal intranuclear inclusion disease.

ObjectiveThe diagnosis of neuronal intranuclear inclusion disease (NIID) is currently based on CGG repeat expansion in the 5′UTR of the NOTCH2NLC gene, or p62-positive intranuclear inclusions in skin biopsy. The purpose of this study is to explore the value of non-invasive pathological findings in urine sediment cells from NIID patients.Materials and methodsTen patients with clinically suspected NIID were enrolled for skin biopsy and gene screening. Morning urine (500 ml) was collected from each patient, and cell sediment was obtained by centrifugation. Urine cytology, including Giemsa staining, p62 immunostaining, and electron microscopic examination, were conducted on cell sediment.ResultsThe main clinical symptoms of 10 patients included episodic disturbance of consciousness, cognitive impairment, tremor, limb weakness, and so on. Cerebral MRI showed that 9 patients had linear DWI high signal in the corticomedullary junction. Genetic testing found that the number of CGG repeat ranged from 96 to 158 in the NOTCH2NLC gene. Skin biopsy revealed that all patients showed p62-positive intranuclear inclusions in 18.5 ± 6.3% of the duct epithelial cells of sweat gland. In contrast, urine sediment smears revealed that only 3 patients had p62 positive intranuclear inclusions in 3.5 ± 1.2% of the sedimentary cells. Ultrastructural examinations showed that intranuclear inclusions were also identified in the cell sediment of the 3 patients.ConclusionUrine cytology may be a new and non-invasive pathological diagnosis technique for some NIID patients, although the positive rate is not as high as that of skin biopsy, which is a sensitive and reliable pathological method for NIID.