Ziehl-Neelsen Staining Method Research Articles

COMPARATIVE ANALYSIS OF TUBERCULOSIS BACTERIA IN SALIVA AND SPUTUM USING ZIEHL-NEELSEN, PAPANICOLAOU AND BLEACH MODIFIED PAPANICOLAOU INDUCED FLUORESCENT MICROSCOPY

Background: This research contrasted the identification of Mycobacterium tuberculosis in saliva and sputum samples using Ziehl-Neelsen, Papanicolaou, and Bleach Modified Papanicolaou fluorescent microscopy.
 Methodology: Sputum and saliva samples were collected in a clean sterile, leak-proof, wide-mouth containers early in the morning. Each saliva and sputum sample was further divided into 3 groups, Group A: sputum stained by Ziehl-Neelsen staining method, Group B: sputum smears stained by Papanicolaou staining method, Group C: sputum smears stained by bleach modified Papanicolaou staining method. saliva and sputum infected by Papanicolaou and observed under a Fluorescent Microscope for tubercle bacilli..
 Results: The results obtained from saliva stained smears showed 63.3% of cases positive in both BMP and Pap stained saliva samples and 46.6% of cases positive in ZN stained saliva samples. Further grading of the individual staining method was carried out. Scant bacilli; 20% in Pap and 10% in BMP, 1+; 30% in Pap and 33.3% in BMP, 2+;10% in Pap and 6% in BMP and 3+-3.3% in Pap and 13% in BMP. ZN stained saliva samples scant bacilli; 56.6%, 1+; 13%, 2+; 10% and 3+; nil. Sputum samples stained by BMP, Pap, and Zn showed 100% positive results (30/30). But the grading differed among individual staining techniques.
 Conclusion: In our study sputum samples showed 100% positivity in comparison to saliva samples with 63.33%. Though the results of the saliva sample were lower than that of the sputum samples, the BMP method used in our study is first of its kind to the best of our knowledge. It is a simpler, convenient, and economical method that can be used in routine diagnosis of TB.

Intestinal protozoan and helminthic infections among hemodialysis and cancer patients.

Intestinal parasitic infections (IPIs) can be a severe threat to immunocompromised patients. This is particularly true for those undergoing chemotherapy and hemodialysis. The present research is aimed at identifying intestinal parasites that might be present in immunocompromised patients. In this cross-sectional study 1040 stool samples were collected from March to September 2017. Six hundred and forty-one stool samples from immunocompromised patients (279 samples from hemodialysis patients and 362 samples from chemotherapy patients) and 399 samples from the control group were collected in Guilan province, Iran. The samples were tested by direct, formalin-ether methods for protozoa and ova of intestinal parasites and Ziehl-Neelsen staining methods for coccidian parasites such as Cryptosporidium species. The overall parasitic infection rate was highest (15%) in hemodialysis patients and 11.3% in chemotherapy patients, whereas the lowest rate was observed (7.3%) in the control group. The infectivity rates were statistically significant (P = 0.008) when compared with the control group. The parasites found were Blastocystis hominis (8.9% of the cases), Entamoeba coli (1.6%), Iodamoeba butschlii (0.8%), Endolimax nana (0.6%), Chilomastix mesnili (0.5%), Strongyloides stercoralis (0.5%), and Taenia species (0.15%), whereas Giardia lamblia was detected only in the control group. There was not a correlation between prevalence of parasites with age or education levels of the infected individuals. Results of the present study suggest that periodic stool examinations in special parasitological laboratories should be included as part of routine and general medical care.

A New Perspective on the Status of the Intestinal Parasitic Infections in the Rural Areas of Fars Province South of Iran

Parasitoses are among the most important problems of most countries especially developing countries. We aimed to detect the situation of intestinal parasitic infections in the Farashband district in Fars Province South of Iran and identify influential factors in the escalation of parasitic diseases and to reduce them. Overall, 1009 participants from the age of 6 months to 90 years were selected from 3 cities and 15 villages of Farashband district, Fars Province South of Iran from 2015 to 2016. Parasitological methods such as the direct assay method, formalin-ether concentration method, and zinc sulfate flotation were used for diagnosis of worm eggs, cysts, and protozoa trophozoite. Susceptible and protozoan positive samples were stained using the Trichrome staining method. The modified acid-fast staining procedure was conducted for diarrheal samples and the results were used for diagnosis of coccidia. Overall, 313 subjects were infected with at least one intestinal parasite (pathogenic and nonpathogenic). Helminthes infection and protozoan infection were observed in 9 (0.9%) and 304 (30.13%) participants, respectively. Fecal samples of 34 patients with diarrheal feces were used to prepare smears for further examinations using the Ziehl-Neelsen staining method. Examinations showed no infection with coccidia. Helminthes infection has decreased drastically but protozoan infection is still considered a health issue in this region. It is possible to reduce parasitic infections through proper measures such as increasing public awareness and education the public, especially children on health problems with education courses.

Comparison of LJ Medium and BACTEC MGIT 960 Culture System for the Diagnosis of Tuberculosis

Introduction: Sputum negative pulmonary Tuberculosis (TB) is a major public health problem. So, the emergence of new techniques for a more precise and rapid microbiological identification of Mycobacterium tuberculosis in clinical samples is of great importance to improve the management of TB. Aim: To determine and compare the sensitivity and turnaround time for Mycobacterium tuberculosis detection by the BACTEC Mycobacteria Growth Indicator Tube (MGIT) 960 system, Lowenstein Jensen (LJ) medium and Ziehl-Neelsen (ZN) staining. Materials and Methods: An Institution based, observational, cross-sectional study was conducted at Rajendra Institute of Medical Sciences (RIMS), Ranchi, Jharkhand, India, from July 2013-March 2016. Sputum, pericardial fluid, pleural fluid, peritoneal fluid, pus and endometrial tissue samples were collected from 80 patients of suspected TB cases. All were Acid-Fast stained by ZN staining method and cultured on solid culture LJ medium and on liquid medium (MGIT). Data was analysed using Statistical Package for the Social Sciences (SPSS) software, Version 20.0 (SPSS Inc, Chicago, IL, USA). Fisher’s-Exact test was used to show association of categorical variables. Non-parametric Mann-Whitney U test was used to show median difference of non-normally distributed continuous variables of two groups. Results: Out of the 80 samples, 41 cases were positive by either of the all methods. The positive specimen for ZN staining, LJ media and MGIT were 21, 29 and 41 cases respectively. The mean Time To Detection (TTD) was shorter for MGIT system than LJ media. Both LJ medium and MGIT 960 detected all cases of sputum smear positive cases and in addition significantly higher number than ZN stain in sputum smear negative cases. MGIT 960 detected significantly higher number of cases of sputum negative cases than LJ Media. The mean TTD was also significantly shorter in case of smear positive cases than the smear negative cases by both the solid and liquid culture mediums. Conclusion: The use of liquid media (MGIT) is more accurate and rapid method for the diagnosis of TB. The combination of more than one method is also highly recommended for rapid detection and early treatment of TB.

Parasitological detection and associated risk factors of Cryptosporidium in bovines of Karnataka state

The prevalence of Cryptosporidium in cattle and buffalo calves of less than one month age was studied in Karnataka state using modified Ziehl-Neelsen staining method. Associated risk factors viz., age, sex, breed, consistency of dung, rearing systems and season were evaluated for their influence on the prevalence of Cryptosporidium infection. An overall prevalence of 12.20% (40/328) of Cryptosporidium infection was observed with a prevalence rate of 12.84% in cattle calves and 10.91% in buffalo calves. Age wise, three week-old animals were infected more (16.95%) followed by the four week (13.27%), one week (6.90%) and two week (4.41%) old animals. Female animals (13.14%) showed more prevalence than male animals (11.11%). Among the breeds, crossbred animals (15.08%) were infected more compared to non-descript animals (7.75%). Higher prevalence of infection was noticed in diarrhoeic samples (16.41%) followed by formed (10.31%) and semisolid dung samples (7.77%). Rearing system was correlated for its effect on its prevalence rate, in which the free-range animals (12.61%) had a slightly higher prevalence than that of intensively reared animals (11.32%). Season-wise, the higher prevalence of infection was noted in monsoon (14.74%) followed by winter (11.48%), autumn (11.38%) and summer (10.20%).

The Investigation of Mycobacterium paratuberculosis by Serological and Cultural Methods In Raw Milks Retailed in Kayseri

Bu çalışmada Kayseri ilinde satışa
 sunulan sokak sütlerinde Mycobacterium
 avium subsp. paratuberculosis
 (MAP)'in varlığı konvansiyonel ve serolojik yöntemlerle araştırılması amaçlandı.
 Araştırmada Mayıs 2010 – Ekim 2011 yılları arasında Kayseri’nin ilçelerinden
 farklı satıcılardan 200 çiğ inek süt örneği toplandı. Her bir süt numunesi
 direkt mikroskopi, kültür ve serolojik yöntem olmak üzere üç aşamada
 değerlendirdi. Direkt mikroskopide süt örneklerinden hazırlanan preparatlar
 Ziehl-Neelsen boyama metodu ile boyandı. Ayrıca besiyerlerinden izole edilen
 şüpheli kolonilerin boyamasında da aynı yöntem kullanıldı. Etken izolasyonu
 amacıyla süt numunelerinin bakteriyolojik ekimleri gliserinli ve gliserinsiz
 Löwenstein-Jensen ve Watson-Reid agara yapıldı. İdentifikasyon için katalaz,
 niasin, nitrat redüksiyon, pirazinamidaz ve üreaz testleri kullanıldı. Sütteki
 antikorların araştırılmasında Mycobacterium
 paratuberculosis Enzyme-Linked ImmunoSorbent Assay (ELISA) kitinden
 yararlanıldı. İncelenen örneklerde üç yönteminin kullanılması sonucu sadece 1
 numunede (% 0.5) MAP tespit edildi. Sonuç olarak, Kayseri ilinde satışa sunulan
 sokak sütlerinde paratüberkülozun yüksek oranda bulunmadığı belirlendi. Ancak
 bir örnekte etkenin tespiti infeksiyonun önemi göz önünde bulundurulduğunda
 halk sağlığı açısından önemli risk oluşturabileceği kanaatini ortaya koymuştur.

Parasitological Prevalence of Cryptosporidium infection in calves, chickens and human in Behera province

Cryptosporidium is a cosmopolitan intestinal protozoan parasite that belongs to the phylum Apicomplexa and infects a broad range of vertebrate hosts including livestock, human and birds. This current study was aimed to investigate the prevalence of Cryptosporidium spp.infection in young calves, human and chickens, in five districts covering five different geographical locations of the Behera province. In doing this, a total number of 310 faecal samples from calves (n= 148), human being (n=112) and chicken (n=50) were collected, stained using the modified Ziehl-Neelsen staining method, and were examined microscopically for Cryptosporidium spp. oocysts. Correlations between prevalence rates and related parameters such as species, age, gender and faecal consistencies were also analyzed. Results indicated that 43.2%, 16.1% and 6%, of calves, human and chicken, were found infected with Cryptosporidium spp., respectively. Statistically significant variable rates of infection were also detected across six surveyed districts of the Behera governorate, with the notable absence of infection from chicken in Abo Hummus and Rahmaneyah districts. Statistically significant variable rates of infection were also characterized in calves when species, age, sex, and faecal consistency were analyzed; buffalo >4 weeks calves, male and calves with formed faeces have higher infection rates than their counterparts of cattle Cryptosporidium spp. in the livestock and chicken living in close proximity to human population necessitates better surveillance and control measures to protect vulnerable animal and human populations.

Histopathological changes of brucellosis in experimentally infected guinea pig

Background: Brucellosis, a chronic infectious and zoonotic disease, is endemic in many countries of the world including Bangladesh, that affects humans and animals, leading to significant impact on public health and animal industry. There are several reports on seroprevalence, risk factors, molecular, epidemiological and review of brucellosis in human and animals but least of histopathological reports in Bangladesh. As laboratory animal species guinea pigs are the most susceptible to Brucella infection in comparison to mice, monkeys, rats and sheep. Hence, this study was undertaken to determine the histopathology of brucellosis in guinea pig. Methods: After collection of aborted bovine foetal membranes from the Central Cattle Breeding and Dairy Farm (CCBDF), Savar, screened with modified Ziehl-Neelsen staining method and inoculum (10 CFU/2ml) prepared from positive samples were inoculated into each of ten guinea pig. The guinea pigs were sacrificed after showing positive results from the serum in Rose Bengal test and from blood in rapid kit test 3 weeks post inoculation. Then the gross and histopathological lesions were observed in the liver, spleen, heart, lungs, and kidneys. Results: Liver showed congestion, haemorrhage and fatty changes and granuloma formation with infiltration of macrophages. However, only caseous necrosis of splenic follicle was observed in spleen. The major findings in lungs of guinea pig were granuloma, haemorrhage and necrosis of lung parenchyma and mononuclear cellular infiltration. The heart revealed haemorrhagic endocarditis and monocytes infiltration. A variable degree of multifocal accumulation of mononuclear cells infiltration and congestion were found in kidneys. Conclusions: It is concluded that the histopathological changes caused by Brucella spp. are similar to those observed in humans with brucellosis. The animal models, particularly the guinea pig, can be used to study the detail pathogenesis of this organism.

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.

PO 8414 EVALUATION OF MYCOBACTERIUM TUBERCULOSIS COMPLEX (MTBC) CULTURE METHODS IN MYCOBACTERIUM AFRICANUM-ENDEMIC REGION OF WEST AFRICA

BackgroundWith the endemic Mycobacterium africanum (Maf), West African laboratories use glycerol and pyruvate in separate LJ cultures (LJG and LJP) for isolation of MTBC. The aim of this work is to evaluate if combining both glycerol and pyruvate in a single LJ medium (LJGP) will lead to comparable growth characteristics and time to detection in comparison to LJG, LJP and MGIT 960.MethodTotal of 118 smear-positive sputum samples were processed using 4% NaOH-NALC decontamination method. The decontaminated samples were inoculated parallel on LJG, LGP, MGIT 960 and LJGP. Positive cultures were confirmed using Ziehl-Neelsen staining method. MTBC identification was done using the Capilia TBNeo kit and spoligotyping used for speciation.ResultsThe recovery rate for LJG, LJP, LJPG and MGIT was found to be 73.7% (87/118), 82.2% (96/118), 83.9% (99/118) and 93.2% (110/118) respectively. No significant agreement was observed between the LJPG and MGIT 960 with Kappa values of −0.105 (p-value=0.199). However, there was significant agreement between LJGP and LJG and LJP with Kappa value of 0736 (p-value<0.001) and 0.756 (p-value<0.001), respectively. There were 70 Euro-American, 34 Maf, 9 East-Asian, 2’Indo-Oceanic, 2 East-African-Indian and 1 M. Bovis. LJGP have better Maf recovery rate, 85.3% (29/34) in comparison to MGIT 960, 79.4% (27/34), LJP, 76.5% (26/34) and LJG, 61.8% (21/34). Seven of the 8 MGIT negatives that were LJPG positive were M. africanum and 1 M. bovis.ConclusionLJPG has a better detection rate and time to positivity compared to LJG and LJP and was shown to have a better Maf recovery than other LJ methods and MGIT 960. It is evident that LJGP is a promising culture tool for Maf-endemic West African countries that will not only increase MTBC recovery rate in combination with MGIT, but also leads to better detection of Maf.

Pulmonary Tuberculosis and Associated Factors Among Diabetic Patients Attending Hawassa Adare Hospital, Southern Ethiopia.

Background:Developing countries have a high burden of Tuberculosis (TB); although it is considered as a disease of the past in most developed countries. The end TB strategy was predicted to stabilize or drop the incidence of TB. However, the rising of the prevalence of immune-related diseases like Diabetes Mellitus (DM) are challenging the TB control strategy in high TB burden region. The objective of this study was to determine the prevalence and associated factors of pulmonary tuberculosis in DM patients attending Adare Hospital, south Ethiopia.Methods:A cross-sectional study was carried out on 207 randomly selected diabetic patients at the Adare hospital. A structured pre-tested questionnaire was used during the data collection from participants. Sputum concentration technique followed by Ziehl-Neelsen staining method was used to examine pulmonary tuberculosis. Logistic regression analysis was used to assess the association between various variables and pulmonary tuberculosis. Odds ratios and 95% CI were computed to determine the strength and presence of the association.Results:The prevalence of pulmonary tuberculosis among diabetics was 5.3% [95% CI: (2.2, 8.4)]. Diabetic patients who were underweight [AOR = 9.94, 95% CI: (1.51-80.89)], had more than 10 years duration with DM [AOR = 7.03 95% CI: (1.357, 73.6)], Alcohol drinking habit [AOR = 12.49, 95% CI: (3.28, 77.94)], and history of contact with TB [AOR = 5.35, 95% CI: (1.1-39.12)] were the factors positively associated with pulmonary TB infection while being HIV seronegative had a negative association with pulmonary TB infection [AOR =0.074, 95% CI: (0.001-0.29)].Conclusion:High proportion of pulmonary TB was observed in diabetic patients as compared to the national estimated prevalence of TB in the total population. Duration of patient with DM, being underweight, alcohol consumption habit, and contact history with TB were positively associated with pulmonary TB infection while being HIV seronegative had a negative association with the infection in diabetic patients.

Morphological and molecular identification of Cyclospora species in sheep and goat at Tamil Nadu, India.

A total of 281 fresh fecal samples (65 sheeps and 216 goats) were examined for the presence of Cyclospora species in Tamil Nadu from October 2017 to April 2018. All the faecal samples were examined by direct smear method, saturated sucrose floatation technique and modified Ziehl-Neelsen staining method and it was confirmed-at genus level with PCR technique. Overall prevalence of Cyclospora infection in-small ruminant was 2.14%. The prevalence of Cyclospora species in sheep and goat was 3.08 and 1.85% respectively. Cyclospora oocyst was observed mostly during winter season (Dec-Feb) with 7.14% followed by summer (March and April) and Northeast mansoon (October and November) with 1.67 and 1.66% respectively. The oocyst detected from faecal sample through conventional faecal examination was confirmed by PCR.

Socio-Behavioral Risk Factors Associated with Cryptosporidiosis in HIV/AIDS Patients Visiting the HIV Referral Clinic at Cape Coast Teaching Hospital, Ghana.

Objective:To identify the socio-behavioral risk factors associated with cryptosporidiosis among HIV/AIDS patients with chronic diarrhea symptoms visiting the HIV referral clinic at Cape Coast Teaching Hospital, Ghana.Methods:A cross-sectional study was conducted among 50 HIV/AIDS patients with recurrent diarrhea. Questionnaires were administered to collect social and behavioral risk factors associated with Cryptosporidium and other opportunistic protozoan parasitic infections in HIV patients. Stool samples were collected for the diagnosis of enteric protozoan pathogens using modified Ziehl-Neelsen and acid-fast staining methods. CD4+ cells counts of study subjects were obtained from patients clinical records. The data obtained were analyzed using Pearson chi-square and multivariate-adjusted statistics tool on SPSS 16 for Windows.Results:Twenty-seven (54%) of the subjects were infected with enteric protozoan pathogens. The prevalences of Cryptosporidium, Cyclospora and Microsporidium infections were 46%, 32% and 16%, respectively. Cryptosporidium infection was significantly associated with drinking water (×2=13.528, p<0.001), Cyclospora was associated with the type of drinking water (×2=14.931, p<0.001) and toilet facilities used by the study subjects (×2=12.463, p<0.01), whiles Microsporidium infection was associated with hand washing behavior (×2=12.463, p<0.01). Enteric protozoans were frequently encountered among subjects with CD4+ T-cell count <200 cells/mm3. However, coinfection of Cyclospora spp & Cryptosporidium spp was not observed in CD4+ cell count <200 and >500 cells/mm3. Multivariate analysis showed that the risk factor for Cryptosporidium infection among HIV/AIDS patients was the source of drinking water (pipe borne water 76.2% prevalence: sachet water 25%; OR=0.10, 95%CI: 0.03-0.39, p<0.001).Conclusion:We report the risk factor for exposure of Cryptosporidium infection among HIV/AIDS patients for the first time in Ghana. The contamination of drinking water by protozoan parasites should be a public health concern. These results provide the stepping block to understand the transmission dynamics of Cryptosporidium and other opportunistic pathogens in HIV/AIDS infected patients in Ghana.

Cytodiagnosis of tuberculosis using modified bleach method on lymph node aspirates.

Background:The conventional Ziehl–Neelsen (ZN) staining method for demonstration of acid-fast bacilli (AFB) by direct microscopy plays a key role in the diagnosis of tuberculosis. Its major disadvantage is low sensitivity.Aims:The present study was undertaken to emphasize the role of bleach concentration method over conventional ZN smear for detection of AFB in fine-needle aspiration (FNA) material of clinically suspected tuberculous lymphadenitis.Settings and Design:Tertiary care teaching hospital, cross-sectional design.Materials and Methods:FNAs were done on 150 patients with clinical suspicion of tuberculosis presenting with lymphadenopathy. Smears from the aspirates were processed for routine cytology and the conventional ZN method. The remaining material in the needle hub and/or the syringe was used for the bleach method. The significance of the bleach method over the conventional ZN method and cytology was analyzed.Results:Among the 150 aspirates, 57% (85/150) were indicative of tuberculosis on cytology, 37% (55/150) were positive for AFB on conventional ZN method, and the smear positivity for AFB increased to 41% (62/150) on bleach method.Conclusion:The bleach method is simple, safe and inexpensive. It improves the microscopic detection of AFB and can be a useful contribution to routine cytology.

Epidemiological Profile of Intestinal Parasitic Infection among Preschool and School Children Living in a Rural Community in Senegal: A Cross Sectional Survey

Introduction: Intestinal parasitic infection (IPI) remains major public health problem in worldwide, being most prevalent in developing countries. Mass drug administration with Mebendazole is an effective strategy for the control of these diseases in pre-school and schoolchildren. The aim of this study was to assess the prevalence of intestinal parasitic infections and established associated risk factors among pre-school and schoolchildren living in rural area in Senegal. Materials and method: A cross sectional survey was carried out between Novembers to December 2014. Children less than 10 years were randomly selected using a systematic random sampling method. Stool samples were examined by light microscopy for the detection of parasite using direct examination, Modified Ritchie Method and the modified Ziehl Neelsen staining method. Results: Among 392 children selected in the study, 137 were found with at least one intestinal parasite representing an overall prevalence of 34.95% [CI 95% (29.3–41.3)]. Protozoan infection was more frequent than helminthic infection (93.4% versus 2.2% respectively). Association between helminths and protozoa was 4.4%. Intestinal parasites identified were Giardia intestinales (72.48%), Entamoeba coli (13.76%), Blastocystis hominis (3.67%) and Hymenolepis nana (1.83%). Prevalence was higher among children aged over 5 years old (48.4%) (aOR= 5.39; 95% IC (2.06–13.9); p=0.001). Children living in family with more than 10 individuals and those drinking unsafe water were found most infected respectively 37.5%) (aOR= 1.24; 95% CI (0.65–2.38); p=0.51) and (42.3%) (aOR=1.45; 95% CI (0.83–2.52); p=0.18). Prevalence was higher in children with underweight 36.3% [aOR= 1.53; 95% IC (0.88–2.67); p=0.13). No correlation was found between sex and intestinal parasitic infection. Conclusion: This study showed that intestinal parasites remain prevalent in pre-school and schoolchildren with a changing of the epidemiological profile. Protozoan infection was more frequent than helminthic infection. Preventive measures including mass drug administration with Albendazole and water supply could reduce the prevalence of these diseases.

Determination of Cryptosporidium spp. in Van cats by nested PCR

The present study was conducted to investigate the presence of Cryptosporidium spp. agents in cats from the Turkish Van Cat Shelter at YüzüncüYıl University by a modified Ziehl-Neelsen staining method and nested PCR. Individual stool samples were obtained from 30 adult females, 30 adult males and 40 kittens – a total of 100 Van cats were analyzed in the study. A simplified formol-ether concentration method was applied to all samples. The samples were then examined microscopically by a modified Ziehl-Neelsen staining method. As a result of the staining, Cryptosporidium spp. oocysts were identified in stool samples of 3 kittens in the microscopic examination. After that, PCR and nested-PCR were conducted with suitable primers. Nested PCR identified 5 kittens (5%) as positive. As a result, it was concluded that nested PCR was a superior diagnostic method for Cryptosporidium diagnosis compared with staining methods and that infected cats could be a health hazard for other cats and individuals, since Cryptosporidium spp. agents infect via the faecal-oral route. Therefore, we believe it is necessary to raise the awareness of people in contact with cats..

Molecular Characterization of Cryptosporidium Species Isolated from Cattle in Southwest of Iran

Background: Cryptosporidium spp. is an ubiquitous intracellular protozoan parasite that affect a wide range of vertebrates like humans and livestock. Objectives: The current survey was aimed to determine the prevalence and molecular characterization of Cryptosporidium spp. in cattle of Ahvaz city, southwest of Iran. Methods: In total, 240 cattle fecal specimens were collected from 5 geographical regions of Ahvaz city and microscopically tested for the presence of Cryptosporidium spp. oocysts. Then, all positive samples were examined by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) to detect and genotyping of Cryptosporidium spp., respectively. Finally, the obtained results were sequenced. Results: The overall prevalence of Cryptosporidium infection by modified Ziehl-Neelsen staining and nested-PCR methods were 2.1% (5/240) in cattle. Digestion of secondary PCR products using the RFLP method and employing SspI and VspI enzymes, revealed C. parvum pattern in all positive cases and also confirmed by sequencing. Conclusions: Current finding suggests that C. parvum is the main species in cattle of Ahvaz city and could be considered as an important reservoir for zoonotic infection.

COMPARATIVE ANALYSIS OF M. TUBERCULOSIS DETECTION BY DIFFERENT HISTOLOGICAL METHODS IN LUNG TISSUE WITH PULMONARY TUBERCULOMA WITH DIFFERENT ACTIVITY OF SPECIFIC INFLAMMATION

Relevance. Modern histological diagnostics of tuberculosis with using of histobacterioscopy is based on the application of Ziehl-Neelsen method, fluorescent and immunohistochemical methods. And it is mainly used for extrapulmonary forms of tuberculosis. From a scientific and practical standpoint, it is advisable to use these methods in a combined.&#x0D; Objective. To conduct a comparative analysis of the detection of M. tuberculosis and their antigens by various histological methods in pulmonary tissue with tuberculoma at different activities of a specific inflammatory process.&#x0D; Materials and methods. The material for the study was 34 cases of resected parts of the lungs with tuberculoma, two groups were formed: the main group – 19 cases with a high degree of activity of specific inflammation; the comparison group – 15 cases with moderately or low activity. Ziehl-Neelsen staining, fluorescent and immunohistochemical methods were applied. The number of infected cells and extracellular bacilli was determined according to the working gradation: rarely (1-5), moderate (6-15), many (more than 16) bacteria in the field of view of the microscope. At the immunohistochemical examination it was determined the type and relative number of cells with clear positive results.&#x0D; Results. All methods of histological diagnosis revealed a free located MBT in the necrotic nucleus and capsule of tuberculoma, and in alveolar spaces at a distance. The greatest number of free located mycobacteria was detected with using fluorescent method and immunohistochemical examination. The Ziehl-Neelsen showed a significant predominance of observations of a moderate amount of infected alveolar macrophages in the main group and a small number of infected macrophages in a group with a moderately or low activity of the inflammatory process (p&lt;0,01). At the progression of tuberculosis, a large number of macrophages with a positive staining to MBT antigens (p&lt;0,001) was more often detected. Highest indicators of diagnostic standards are established for fluorescent and immunohistochemical methods.&#x0D; Conclusion. A combined histological study provided the most complete data on the presence, localization and distribution in the affected tissue of both whole structures MBTs and their antigens at the different activity of specific inflammation. Immunohistochemical study showed that the accumulation of MBT antigens is observed in different types of macrophages and monocytes cells.

Surface Energetic Effects of Human Immunodeficiency Virus Co-Infection on Mycobacterium Tuberculosis-Macrophage Interaction Mechanism

The mechanism of mycobacterium tuberculosis macrophage interactions was studied using surface energetics tool in determining the interaction processes. The surface interfacial energies were explained using van der Waals concept of particle interactions. Twenty samples each of infected and uninfected sputum were collected and screened to determine the infection status using GeneXpert and Ziehl-Neelsen staining method. The absorbance, ā, values of each specimen, for wavelength range of 230-950nm were measured using digital Ultraviolet Visible Spectrophotometer. Matlab software tools were used in the mathematical analysis of the data generated from the absorbance values. The combined Hamaker coefficients were obtained using the values of the dielectric constant with the Lifshitz Equation. The harmonized Hamaker coefficients A132har and the absolute combined Hamaker coefficients A132abs of infected samples were deduced. The values of A132abs for mtb infected sputum and A132abs for mtb/HIV co-infected sputum are 0.21631x10-21Joule and 0.18825x10-21Joule respectively. This result reveals that the surface energy of the macrophage is less than the surface energy of the mtb, mtb/HIV while the surface energy of the mtb/HIV macrophage is less than that of mtb macrophage. HIV has the tendency to reduce the interaction energy with the consequences of increased viral loads and decreased immune systems. The interaction energy is reduced by 13% confirming adverse effects observed in HIV patients suffering from Tb. Negative Hamaker coefficient of -0.22669x10-19mJ/m2 was calculated indicating that separation of mtb is practical. The effect of the infection can only be abated if additive(s) in form drug is added that can render A132 negative. Thus, the value of Hamaker constant of the sputum A33 was required for repulsion to occur mathematically derived as A33≥0.9527x10-21Joule which satisfies this condition for negative A132abs.

Molecular characterization of bovine Cryptosporidium isolated from diarrheic calves in the Sudan.

Cryptosporidiosis is a common protozoan infection causing morbidity and mortality in young cattle and may be zoonotically transmitted to humans. So far, there is no data available on the presence of Cryptosporidium spp. in the Sudan. The aim of this study was to isolate, identify, and genotype Cryptosporidium oocysts sampled from diarrheic calves housed at different farms in three different municipalities in Khartoum State (Khartoum, Khartoum North, Omdurman). A total of 149 fecal samples were evaluated microscopically for the presence of Cryptosporidium oocysts using the modified Ziehl-Neelsen staining method and 87 (58.3%) samples tested positive. Positive and negative samples were further analyzed by nested PCR targeting the SSU rRNA region. Positive samples were subjected to restriction enzyme analysis of PCR amplicons (PCR-RFLP). Nested PCR identified Cryptosporidium DNA in 53 samples (35.5%); restriction digestion of the PCR products revealed the presence of C. parvum (73.5%), C. ryanae (13.2%), C. andersoni (7.5%), and C. bovis (1.8%). Species distribution was clearly related to age with C. parvum being the predominant species in dysenteric pre-weaned calves. Sequencing of three genes (SSU rRNA, COWP, and GP60) for three C. parvum isolates originating from the three different municipalities showed that all belong to C. parvum subtype family IId. Based on data obtained by GP60, sequencing the two C. parvum isolates from Khartoum and Omdurman represent subtype IIdA18G1, whereas oocysts isolated in Khartoum North belong to subtype IIdA19G1. The observed genotypes are zoonotic and thus C. parvum in calves is potentially a health risk to humans in Khartoum State, Sudan. To the best of our knowledge, this is the first reported attempt to characterize Cryptosporidium isolated from cattle in the Sudan.