Ziehl-Neelsen Acid-fast Staining Research Articles

Comparison of microscopic and xpert MTB diagnoses of presumptive mycobacteria tuberculosis infection: retrospective analysis of routine diagnosis at Cape Coast Teaching Hospital

IntroductionTuberculosis is a global health problem that causes 1. 4 million deaths every year. It has been estimated that sputum smear-negative diagnosis but culture-positive pulmonary TB diagnosis contribute to 12.6% of pulmonary TB transmission. TB diagnosis by smear microscopy smear has a minimum detection limit (LOD) of 5,000 to 10,000 bacilli per milliliter (CFU/ml) of sputum result in missed cases and false positives. However, GeneXpert technology, with a LOD of 131–250 CFU/ml in sputum samples and its implementation is believe to facilitate early detection TB and drug-resistant TB case. Since 2013, Ghana health Service (GHS) introduce GeneXpert MTB/RIF diagnostic in all regional hospitals in Ghana, however no assessment of performance between microscopy and GeneXpert TB diagnosis cross the health facilities has been reported. The study compared the results of routine diagnoses of TB by microscopy and Xpert MTB from 2016 to 2020 at the Cape Coast Teaching Hospital (CCTH).MethodsThe study compared routine microscopic and GeneXpert TB diagnosis results at the Cape Coast Teaching Hospital (CCTH) from 2016 to 2020 retrospectively. Briefly, sputum specimens were collected into 20 mL sterile screw-capped containers for each case of suspected TB infection and processed within 24 h. The samples were decontaminated using the NALC-NaOH method with the final NaOH concentration of 1%. The supernatants were discarded after the centrifuge and the remaining pellets dissolved in 1–1.5 ml of phosphate buffer saline (PBS) and used for diagnosis. A fixed smears were Ziehl-Neelsen acid-fast stain and observed under microscope and the remainings were used for GeneXpert MTB/RIF diagnosis. The data were analyze using GraphPad Prism.Results50.11% (48.48–51.38%) were females with an odd ratio (95% CI) of 1.004 (0.944–1.069) more likely to report to the TB clinic for suspected TB diagnosis. The smear-positive cases for the first sputum were 6.6% (5.98–7.25%), and the second sputum was 6.07% (5.45–6.73%). The Xpert MTB-RIF diagnosis detected 2.93% (10/341) (1.42–5.33%) in the first and 5.44% (16/294) (3.14–8.69%) in the second smear-negative TB samples. The prevalence of Xpert MTB-RIF across smear positive showed that males had 56.87% (178/313) and 56.15% (137/244) and females had 43.13% (135/313) and 43.85% (107/244) for the first and second sputum. Also, false negative smears were 0.18% (10/5607) for smear 1 and 0.31% (16/5126) for smear 2.ConclusionIn conclusion, the study highlights the higher sensitivity of the GeneXpert assay compared to traditional smear microscopy for detecting MTB. The GeneXpert assay identified 10 and 16 positive MTB from smear 1 and smear 2 samples which were microscopic negative.

First detection of bovine tuberculosis by Ziehl-Neelsen staining and polymerase chain reaction at dairy farms in the Lekok Sub-District, Pasuruan Regency, and Surabaya region, Indonesia.

Bovine tuberculosis (TB) is a zoonotic disease of great public health importance, particularly in Indonesia, where control measures are limited or are not implemented. This study aimed to detect the presence of Mycobacterium pathogens in milk samples from dairy cattle in Pasuruan regency and Surabaya City, East Java, using Ziehl-Neelsen acid-fast staining and polymerase chain reaction (PCR). Milk samples were aseptically collected from 50 cattle in the Lekok Subdistrict, Pasuruan Regency, and 44 from dairy farms in the Lakarsantri Subdistrict, Wonocolo Subdistrict, Mulyorejo Subdistrict, and Kenjeran Subdistrict, Surabaya, East Java. To detect Mycobacteria at the species level, each sample was assessed by Ziehl-Neelsen staining and PCR using the RD1 and RD4 genes. The results of PCR assay from 50 samples in Lekok Subdistrict, Pasuruan Regency showed that 30 samples (60%) were positive for Mycobacterium tuberculosis and two samples (4%) were positive for Mycobacterium bovis, although Ziehl-Neelsen staining did not show the presence of Mycobacterium spp. In the Surabaya region, 31 samples (70.45%) were positive for M. tuberculosis and three samples (6.8%) were positive for M. bovis. Six samples (13.63%) from all PCR-positive samples could be detected microscopically with Ziehl-Neelsen. The presence of bovine TB in this study supports the importance of using a molecular tool alongside routine surveillance for a better understanding of the epidemiology of bovine TB in East Java.

Cryptosporidiosis in HIV-seronegative patients: A case series

Enteric infections by are a frequent cause of diarrhea among immunocompromised and HIV infected patients but there are limited reports of their occurrence in immunocompetent patients. Cryptosporidiosis is a faeco-orally transmitted protozoan infection and is one of the leading causes of chronic parasitic diarrhea. Here we report a case series of cryptosporidiosis from three different patients where two patients are immunocompetent and one is a post renal transplant case. All the patients came with watery diarrhea for which stool samples were collected and examined by modified Ziehl Neelsen acid fast staining (Kinyoun’s method). On microscopic examination of smear, multiple, pink, round to oval acid fast oocysts of 4-6µm diameter were seen against a blue background morphologically resembling . Early diagnosis and prompt initiation of antiparasitic drugs aids in timely treatment and prevention of disseminated cryptosporidiosis.

Molecular evaluation of Cryptosporidium spp. among breeding calves of Lorestan province Western Iran.

Cryptosporidium spp. are opportunistic intestinal protozoans with global distribution and are of great importance as zoonotic protozoans are common to humans and domestic animals, including cattle and calves. Identification and detection of parasite species using precise methods including molecular methods can be an effective step in treating and controlling parasites. This study aimed to investigate the prevalence of Cryptosporidium among breeding calves of Khorramabad city, Lorestan province, Western Iran, using PCR. The faecal samples were taken from 181 healthy and diarrhoeal calves and after the Ziehl Neelsen Acid-fast staining and microscopic evaluation, the genomic DNA was extracted for molecular evaluations. To detect Cryptosporidium species, specific primers targeting the SAM-1 gene of Cryptosporidium and a commercial master mix were used for PCR. Out of 181 faecal samples of breeding calves in Khorramabad city, 9 samples (5%) were positive for Cryptosporidium spp. using the PCR method. Statistical analysis of the data showed that there was no significant statistical relationship between Cryptosporidium infection of the calves and variables of age, breed, type of water consumption, clinical signs of diarrhoea, and sampling location, while parasite infection had a significant relationship with calf gender so that all Cryptosporidium positive samples were from male calves (p ≤ 0.05). Considering the presence of Cryptosporidium infection, the region's traditional grazing system, and the close relationship between livestock and humans, there is a possibility of human infection in the region. So preventive measures such as periodic animal testing with sensitive and accurate diagnostic techniques including PCR, pharmacological treatment of livestock, water hygiene and the use of industrial grazing instead of traditional grazing to improve the hygiene of food consumed by livestock are recommended.

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.

Detection and pathological role of intestinal protozoa in children.

Intestinal parasites are considered a growing public health problem, being protozoa the main cause of intestinal disease. The objective of our study is to compare the detection of intestinal protozoa by microscopy versus real-time PCR, as well as to determine the most prevalent protozoa in our environment in the paediatric population. An observational longitudinal study was carried out, both by microscopy and real time-PCR in stool samples from children (0- 15years) received from April 2019 to March 2021.Children were classified in two groups according if they had or not had clinical parasitosis. Microscopic examination was performed in all samples using the Ritchie concentration technique with the commercial Mini PARASEP system (Movaco-Grifols®). The presence of Cryptosporidium sp. was evaluated with the modified Ziehl-Neelsen acid-fast stain. The real-time PCR was performed to all samples using the Allplex ™ gastrointestinal parasite panel 4 (Seegene®). During the study period, 500 samples were received, being positive 31 (6.2%) by microscopy and 256 (51.2 %) by PCR. By microscopy, Blastocystis hominis was the most frequently observed (4.8%), followed by Giardia lamblia (1.6%), Dientamoeba fragilis (0.2%) and Cryptosporidium species (0.2%). Regarding the identification by PCR, D. fragilis (35.2%) was mainly identified, followed by B. hominis (28.1%), G. lamblia (7%) and Cryptosporidium sp. (0.8%) without finding clear differences in aetiology according to age. In the case of B. hominis and D. fragilis, there were not differences in the detection of these protozoa between the control group and children with clinical parasitosis (p=0.11). Real-time PCR increases the detection of intestinal protozoa, being underdiagnosed by microscopy, especially D. fragilis, in which PCR is considered the most appropriate method for its detection.

Association between Piscine Mycobacteriosis and Morgellons Disease: A Review of Literature

Piscine mycobacteriosis is a fatal fish illness that affects a variety of species globally. It affects over 200 species of freshwater and marine fish. Several species of Mycobacterium are responsible among them Mycobacterium marinum is the comment. It also affects humans when exposed to contaminated water. In fishes, the symptoms include eroded fins, body surface coated with heavy mucus, changing pigmentation, swelling of abdomen, ulcerative dermal necrosis, and scale loss. In humans, the infection is classified into three clinical groups. Type I is a self-limiting, verruca lesion. Type II is single or numerous subcutaneous granulomas in the presence or absence of ulceration. Type III is deep infections of the tenosynovium, bones, bursa, or joints, resulting in tenosynovitis, osteomyelitis, and septic arthritis. The diagnosis is made by Ziehl-Neelsen acid-fast staining, culture, biochemical reaction, and PCR being the most reliable approach. Piscine mycobacteriosis is treated by antibiotics and vaccination has been considered for its long-term prevention in order to reduce morbidity and mortality. Morgellons disease (MD) is a filamentous dermopathy in which lesions with strange filamentous inclusions appear out of nowhere. Furthermore, formication may accompany dermopathy. The identification of Borrelia spirochetes directly in Morgellons disease patient specimen is constant and repeatable when sensitive and precise detection techniques are utilized. It has been diagnosed by microscopy, histology and molecular diagnostic techniques which are highly sensitive and specific. Morgellons disease is still a myth therefore its treatment is evolving, up to date it has been treated symptomatically.

Alterations in host biomarkers in Cryptosporidium infected goats

Abstract Cryptosporidiosis is the second major cause of diarrheal illness in humans and animals. Despite being a major cause of digestive anomalies in goats of Khyber Pakhtunkhwa (Pakistan), there is a serious dearth of data regarding the infection induced alterations in physiological biomarkers. The present study was conducted to record changes in vital body parameters caused by the cryptosporidiosis. A total of 384 fecal and blood samples were randomly collected from adult goats showing clinical signs of the disease. Fecal samples were examined using the modified Ziehl Neelsen acid fast staining, and were confirmed for the infection using two-step nested PCR assay. A total of 66 animals were included in the diseased group, while 57 were included in healthy animals group. Blood samples were processed for measurement of hematological and biochemical parameters using automatic analyzer. Diseased goats showed significant increase in lymphocytes, eosinophils and packed cell volume (P

The diagnostic yield of closed needle pleural biopsy in exudative pleural effusion: a retrospective 10-year study.

BackgroundPleural effusion is a common presentation in clinical practice. About 40% of exudative pleural effusion is unable to be diagnosed through thoracentesis, and closed pleural biopsy (CPB) is needed. This study was designed to investigate the diagnostic yield of CPB in exudative pleural effusion.MethodsThis was a retrospective 10-year study of patients with unexplained exudative pleural effusion who underwent CPB in two centers. Malignant pleural effusion (MPE) was diagnosed when there was histopathological evidence of pleural tissue, pulmonary tissue, or pleural fluid. Tuberculous pleural effusion (TPE) was confirmed when granuloma or coagulative necrosis was observed in pleural tissue, Ziehl-Neelsen acid-fast staining was positive, or adenosine deaminase (ADA) in pleural effusion was higher than 35IU with clinical symptoms of TB orγ-interferon increased with symptoms of TB.ResultsA total of 644 patients were enrolled, of which 479 were specifically diagnosed (217 patients with TPE and 262 patients with MPE). The sensitivity of CPB in the diagnosis of MPE was 51.5%. Among the pathological types of MPE, lung adenocarcinoma accounted for 77.9% (204/262) of cases, making up the largest proportion. The sensitivity of CPB for diagnosing TPE was 68.7%.ConclusionsCPB has a relatively high sensitivity in the diagnosis of exudative pleural fluid, especially in relation to tuberculous lesions. CPB could provide an alternative technique in clinical practice, especially for basic hospital units without thoracoscopy.

Epidemiological Study on Bovine Tuberculosis in Boran Breeds Cattle at Yabello District, Southern, Ethiopia

A cross-sectional study was conducted at Yabello district, Southern Ethiopia from December 2009 to April 2010 with the aim of an epidemiological survey of Bovine tuberculosis through different diagnostic approach Comparative Intradermal Tuberculin test (CIDT), Questionnaire survey, Abattoir inspection, Bacteriological, and Histopathological examination on animals presenting in the study area. A retrospective study of human TB cases recorded at Yabello Hospital over a six-year (2004-2009) period was also analyzed to determine the significance of the extrapulmonary tuberculosis (EPTB) form of human TB in the district. During the study period, 790 heads of local Boran breed cattle were included among which 540 heads of cattle were subjected for the CIDT test, while the remaining 250 heads were subjected only for abattoir inspection. The prevalence of gross tuberculous lesion positive samples was 16.4%. These samples were again collected for Ziehl Neelsen acid-fast stain and histopathological examination. The result of Ziehl Neelsen acid-fast stain has indicated that, 19.5% positive and 80.5% negative. There was a statistically significant association between the occurrence of the lesions and body condition score (BCS) (χ²=10.903, P=0.006). On the other hand, the result of the CIDT test has revealed 1.3% positive, 7.2% suspected, and 91.5% negative. All selected traits (sex, Age. BCS, and origin of the animal) measured with the PPD test did not show a statistically significant difference among the categories (p>0.05). Besides, a questionnaire survey conducted on 23 households of animal owners indicated that 95.7% had the habit of raw milk consumption. A retrospective study of human TB cases recorded for six years revealed that there was a 13.76% proportion of EPTB forms. The prevalence of BTB in the study area is slightly high when compared with the previous study. However, it needs further investigation and surveillance to explore the control and prevention methods of BTB and its zoonotic importance which requires the awareness of the population on the management of the disease and associated risk factors.

STRUCTURAL ASSESSMENT OF VIOLATIONS OF THE ADAPTIVE MECHANISMS OF ADRENAL CALVESIN THE ACUTE COURSE OF CRYPTOSPORIDIOSIS

The study was the first to assess the adaptation mechanisms disorders of the adrenals of calves in acute cryptosporidiosis. The cryptosporidiosis in patients with symptoms of diarrhea in the farms of the Republic of Tatarstan was diagnosed through formalin-ether sedimentation and following staining of infectious agents by Ziehl-Neelsen acid fast staining method. For specific identification of cryptosporidium in the feces of calves, an enzyme immunoassay kit, “H&R Crypto Rapid Test”, was used to detect cryptosporidium in feces. The agents of cryptosporidiosis of the genus Cryptosporidium were determined in pathological material taken from sick and dead at the age of 5 to 10 days calves. Histological examination of the organs and tissues of calves with the cryptosporidiosis were carried out using generally accepted method of staining of histological sections with hematoxylin and eosin. Structural studies of adrenal cortex and medulla of calves with the cryptosporidiosis revealed significant changes in synthesized hormonal products profiles. The characteristics of intense glucocorticoid biosynthesis were found in the adrenal cortex at the height of disease, as shown by a sharp expansion in zona fasciculate of the cortex, numerous small and large vacuoles in the cytoplasm of spongiocytes and enhanced profiles of capillary net. Intense excretion of glucocorticoids in hemocirculation maximally activated the catabolic processes in the organisms of sick calves. Small number of adrenocorticocytes, the predominance of small hyperchromic nuclei in them and the absence of characteristics of mitotic activity were not edinzona glomerulosa of the cortex that corresponded to the morphological equivalent of retardation of mineralcorticodes biosynthesis. There had been a dramatic increase in cells with characteristics of apotosis among adrenocorticocytes in all zones of the cortex, including the small in width reticular zone. The small catecholamine-synthesizing cells in the medullary area were represented predominantly by small with sharply clarified cytoplasm and small rounded adrenocytes nuclei and by significantly smaller amount of larger noradrenocytes. The found pathological changes in the cortex and medulla of the adrenals characterized the increase of catabolic processes, vascular, metabolic disorders in organisms of calves with the cryptosporidiosis. In view of disorders of adaptation mechanisms and limited plastic resources, a fast death was observed in newborn calves.

Cryptosporidium oocyst shedding in buffalo calves in Haryana: A case study

Bovine cryptosporidiosisis primarily associated with neonatal diarrhoea with higher morbidity than mortality in young calves till they attain immunological maturity. The present investigation relates to a report on the shedding of Cryptosporidium oocyst in two buffalo calves of buffalo farm, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar from 1st days up to 3 month of age at 15 days interval using simple conventional microscopy. By using formol-ether concentration technique followed by modified Ziehl-Neelsen (ZN) acid fast staining, Cryptosporidium oocysts were concentrated and identified. The Cryptosporidium oocysts appeared as reddish pink coloured bodies against a bluish/greenish coloured background at oil immersion using ZN staining kit. The maximum oocyst shedding was observed (2.3 oocyst / field) during 16 to 30 days of age. The oocyst shedding gradually decreased with increase in age afterwards clearly indicating the disease of young buffalo calves.

Cryptosporidiosis in a fire skink (Lepidothyris fernandi) and molecular identification of infecting species

Cryptosporidiosis is an infectious protozoan disease that affects a wide range of animals including reptiles. This is the first report of cryptosporidiosis in a fire skink (Lepidothyris fernandi), an insectivorous reptile commonly found in tropical West Africa. Faecal sample was collected from a fire skink at necropsy for the detection of parasites by faecal sedimentation method, Ziehl-Neelsen (ZN) acid-fast staining, Nested Polymerase Chain Reaction (PCR) and Nucleotide sequencing. Sections of the intestines were also processed for histopathology. Light microscopy revealed the presence of Ophidascarids sp. eggs and Cryptosporidium oocysts. Amplification of the 18S rRNA gene and nucleotide sequencing confirmed Cryptosporidium varanii as the infecting species. Histopathology revealed cellular infiltration and disruption of the epithelial cells along the brush border characteristic of intestinal inflammation.

Cryptosporidium oocyst shedding in buffalo calves in Haryana: A case study

Bovine cryptosporidiosisis primarily associated with neonatal diarrhoea with higher morbidity than mortality in young calves till they attain immunological maturity. The present investigation relates to a report on the shedding of Cryptosporidium oocyst in two buffalo calves of buffalo farm, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar from 1st days up to 3 month of age at 15 days interval using simple conventional microscopy. By using formol-ether concentration technique followed by modified Ziehl-Neelsen (ZN) acid fast staining, Cryptosporidium oocysts were concentrated and identified. The Cryptosporidium oocysts appeared as reddish pink coloured bodies against a bluish/greenish coloured background at oil immersion using ZN staining kit. The maximum oocyst shedding was observed (2.3 oocyst / field) during 16 to 30 days of age. The oocyst shedding gradually decreased with increase in age afterwards clearly indicating the disease of young buffalo calves.

Pathological investigations on an outbreak of avian Tuberculosis (AvTB) in a Turkey flock in Dhaka

A flock of four-week old turkey chicks showed clinical signssuch as fatigue, in-appetence, gasping and eventual death after having a very brief illnesses. Postmortem examinations showed non-mineralized elevated nodules in the lung, costal membranes and inner surface of the abdominal cavity. Histopathologic findings were typical of avian tuberculosis, with necrotic debris in the lung parenchyma. The lesions were found to be surrounded by epithelioid macrophages, lymphocytes, and multinucleated giant cells. Laboratory confirmation was made based on lesions and using Ziehl-Neelsen(ZN) acid-fast stain. The presence of granulomas in the lungs of the infected chicks may suggest that the exposure to the infective agent was via the respiratory route rather than the alimentary route. The case was diagnosed as tuberculus peri-bronchial pneumonia.

First molecular investigation of Cryptosporidium spp. in young calves in Algeria.

To date, no information is available on the prevalence and genetic identity of Cryptosporidium spp. in cattle in Algeria. In this study, 17 dairy farms in the province of Batna, located in the northeast of the country, were visited to collect 132 fecal samples from young calves (< 8 weeks old). Samples were examined microscopically using the modified Ziehl-Neelsen acid-fast staining method, and at least one sample per farm was submitted for molecular analysis. Amplification of a fragment of the small subunit ribosomal RNA gene was positive for 24 of the 61 samples (40%), and sequence analysis identified three species, namely Cryptosporidium bovis (n = 14), C. ryanae (n = 6), and C. parvum (n = 4). The C. parvum IIaA13G2R1 subtype, an uncommon zoonotic subtype, was identified in two isolates from a single farm by sequencing a fragment of the GP60 gene. This is the first report about genotyping and subtyping of Cryptosporidium in calves in Algeria.

Genetic Characterization of Cryptosporidium spp. among patients with gastrointestinal complaints referred to clinical laboratories of Tehran, Iran

Background : Cryptosporidium, an intracellular protozean parasite, is among the major causative agents of gastroenteritis disorders in humans and also causing water-borne and food-borne outbreaks of diarrheal diseases. This study investigated subtypes of Cryptosporidium in patients with gastrointestinal complaints in Tehran, Iran. Methods: A total of 1685 fecal samples were collected from patients with gastrointestinal complaints who had been referred to clinical laboratories Tehran, Iran. The primary diagnosis was established by detection of oocysts using the modified Ziehl-Neelsen acid-fast staining method and following that, the positive microscopically samples were selected for sequence analysis of partial 60 kDa glycoprotein (gp60) gene. Results: Out of 1685 collected samples, 7 (0.4 %) were positive for Cryptosporidium oocysts. Sequences analysis of gp60 gene in seven Cryptosporidium isolates revealed that, two subtype families were identified, IIa and IId. Five (of 7) isolates belonged to the subtype family IIa and remaining 2 isolates belonged to IId. Three sub-types were recognized within the subtype family IIa including IIaA16G2R1 (3/5), IIaA17G1R1 (2/5), while IIdA17G1d was the only subtype within IId subtype family. Conclusion: The predominance of zoonotic subtype families of C. parvum species (IIa, IId) in this study highlights the importance of zoonotic transmission of cryptosporidiosis in the country.

Introducing Miltefosine as an Anti-cryptosporidial Agent in Immunocompromised Mice

Cryptosporidiosis is a clinically significant opportunistic infection causing diarrhea especially among immunocompromised patients includes those with immunodeficiency syndrome and certain immunosuppressed patients undergoing chemotherapy, organ transplant recipients treated with drugs that suppress the immune system, and patients with autoimmune disorders. The aim of this study was to evaluate the potential antiparasitic effect of miltefosine, (a phospholipid drug used for the treatment of visceral and cutaneous leishmaniasis) in immunocompromised (Dexamethasone treated) mice infected with Cryptosporidium. Parasitological examination of feces for oocysts count was performed ten and twenty days after start of treatment. Histopathological examination of intestinal, liver and spleen sections was held. Results revealed no significant reduction in mean Cryptosporidium oocyst number detected in immunocompromised infected group ten days post treatment (1.44%). Twenty days post treatment showed statistical significant reduction (p<0.001) reaching (38.63 %) when compared to the mean oocysts counts of infected untreated immunosuppressed group of mice. Histopathological sections of small intestine, liver and spleen showed several degrees of inflammatory changes before treatment. In treated group with miltefosine no improvements of small intestinal photomicrographs were seen; in contrast, significant improvement was observed in liver and spleen histopathological sections. Ziehl–Neelsen acid-fast stain was originally undertaken for the detection of Cryptosporidium oocysts within intestinal mice tissue. In conclusion, oral administration of miltefosine in vivo showed moderate efficiency against cryptosporidiosis in immunocompromised infected mice.

INTESTINAL PARASITES IN DIABETIC PATIENTS IN SOHAG UNIVERSITY HOSPITALS, EGYPT.

Intestinal parasites usually create benign diseases, though they may induce complications with high morbidity and mortality to the immunocompromised, including diabetic patients. The study detected the prevalence of intestinal parasitic infections in diabetic patients, comparing to non-diabetic controls and other parameters. A total of 100 fecal samples were collected from diabetic patients at the outpatient clinic of Sohag University Hospitals and another 100 from cross matched controls. The samples were examined macroscopically and microscopically by direct smear and different concentration methods then stained by Modified Ziehl-Neelsen Acid fast stain. Glycated hemoglobin (Hb Alc) was measured to detect DM controlled patients. The data were organized, tabulated, and statistically analyzed. Intestinal parasites were found in 25 (25%) cases out of 100 patients in diabetic group and 7(7%) cases out of 100 controls with high significance (P<0.001)). In the diabetic group, Giardia lamblia was detected in 22 cases (22%) and 5 (5%) among controls, Entamoeba histolytica in 7 cases (7%) and 3 (3%) among controls, Hymenolypis nana in 5 cases (5%) and 3 (3%) among controls, Entamoeba coli in 8 patients (8%), Entamoeba hartmanni in 3 cases (3%), Dientamoeba fragilis in a case (1%), Cryptosporidium parvum in 5 cases (5%) and microsporidia in 3 cases (3%). But, E. coli, E. hartmanni, D. fragilis and C. parvum nor microsporidia were detected in controls. The rate of G. lamblia in DM patients compared to controls was high significant (P<0.001). Hymenolepis nana was 5% (5 cases) in diabetic patients compared to 3% (3 cases) in controls. Residence and sex differences were not significant, while age, >10 years showed the highest prevalence (P< 0.003), type I infection rate was significantly higher than type II (P<0.001). DM control was also significantly affected the infection rates (P<0.007 in type I and P< 0.01 in type II).

Comparison of chest x-ray findings of smear positive and smear negative patients with pulmonary tuberculosis.

Background:Tuberculosis is a chronic pulmonary infectious disease that has affected one-third of the people in the world. It causes nine million new cases and two million deaths per year. Chest radiography associated with Ziehl-Neelsen acid-fast staining procedure significantly helps the diagnosis of pulmonary tuberculosis (PTB). Chest radiography can help the diagnosis of tuberculosis in patients with a negative smear sample result that is mainly diagnosed with delay.Objectives:In this study, chest X-ray findings of PTB were compared in two groups of smear positive and smear negative patients.Patients and Methods:In this retrospective descriptive-analytical study, 376 patients who had been confirmed with PTB were referred to Birjand Health Care Center from 2001 to 2006. Out of the 376 patients, 100 patients with a positive smear based on WHO criteria were selected. In addition, among negative smear patients, 100 were selected in whom similar demographic characteristics with positive smear patients were seen. All of them had undergone chest radiographies that were then interpreted by two expert radiologists independently. Moreover, all patients’ sputa were examined by an expert laboratory technician at the reference laboratory of the health center. The obtained data were analyzed by means of frequency distribution table and descriptive statistics using SPSS (version 15) and Chi-square statistical test.Results:Except reticulo-nodular infiltration, the relative frequency of other radiographic findings in positive smear patients were more than negative smear patients; and only differences in calcification variables, mediastinal widening, patchy infiltration and hilar adenopathy were statistically significant (P < 0.05).Conclusions:Based on the results of this study, although radiographic findings are not diagnostic in PTB, they are helpful if the assessment associates with the view of clinical manifestations and sputum smears.