Modified Ziehl-Neelsen Staining Research Articles

Modified Ziehl–Neelsen staining for Mycobacterium leprae

Modified Ziehl–Neelsen staining for Mycobacterium leprae is a time-tested technique in the evaluation of leprosy. The staining helps to classify clinical types of leprosy into those with low and high bacillary load. It is also useful in assessing the response to therapy, identifying defaulters, and differentiating relapse from late reversal reactions. In this net educational video, we demonstrate the technique of taking an ear lobe smear and performing the modified Ziehl–Neelsen staining for acid fast bacilli.

Intestinal Parasite Infections and Associated Risk Factors among Pre-School Aged Children in Kibera Informal Settlement, Nairobi, Kenya

Infections with intestinal parasites are a major public health problem in children in developing countries like Kenya. School going children are considered at most risk and are included in school-based de-worming program. Less focus is given to pre-school children and information is scarce about intestinal parasitosis among this age group. In this study, we determined the prevalence and intensity of protozoa and helminth infections, and associated risk factors in an informal settlement. A community based cross-sectional study was conducted from October 2016 to January 2017 among 406 children aged 2-5 years in Kibera informal settlements in Nairobi County, Kenya. Structured interviewer-administered questionnaire was used to collect sociodemographic information and data on associated factors. Stool samples were examined microscopically using formal ether concentration, iodine wet mounting, modified Ziehl-Neelsen staining, and Kato-Katz methods. Multivariable logistic regression analysis was used to identify risk factors associated with intestinal parasites. The overall prevalence of any helminth and protozoa infections was 13.1% 53/406) and 22.4% (91/406) respectively. The predominant parasites were Giardia lamblia (13.8%), Ascaris lumbricoides (11.3%), Entamoeba histolytica/dispar (9.4%), Trichuris trichiura (3.9%), Entamoeba coli (1.5%) and hookworm (0.2%). Prevalence of co-infection with any helminths or protozoan was 2.7%. About 10.8% (44/406) and 20.7% (84/406) children were infected with single species of helminth and protozoan parasites. All helminth infections were light, with a mean intensity of 592 egg per gram. Intensity of any protozoan infections was heavy 62.6% (57/406). Dirt floors in the household (aOR = 2.22, p = .046), dirty toilets (aOR = 2.33, p=.014), water from communal taps (aOR = 0.27, p=.019), parent's education level (aOR=0.27, p=.032) and parent's earning (aOR =3.34, p=.007) were factors significantly associated with intestinal parasites. The study found both helminth and protozoan parasites to be prevalent among pre-school aged children in Kibera. Intervention measures including education on the improvement of hygiene and health, socio-economic conditions, sanitation, and provision of safe drinking water could reduce the prevalence of these infections.

Molecular subtype analysis of Cryptosporidium spp. in sheep of Babylon province, middle Iraq

The aim of this study was to investigate the zoonotic potential and genetic diversity of Cryptosporidium species and subtype of C. parvum & C. hominis in sheep. A total of 180 fecal samples were collected from four sites in Babylon province, tested by modified Ziehl-Neelsen stain and 50 positive samples were exposed to the Nested PCR reaction. Four species (C. parvum, C. hominis, C. ubiquitum and C. andersoni) were identified based on sequence analysis of the heat shock protein 70 (Hsp70). Molecular phylogenetic analysis of the gp60 genetic sequence of 10 C. parvum, and 10 C. hominis isolates revealed diagnosis of four subtype included IIdA17G1a, IIdA17G1, IIdA18G1, IIdA21G1 and two subtypes of C. hominis, (IbA21G2 and IbA13G3). The result suggest that the sheep may be considered as reservoir host of IId (C. parvum) and Ib (C. hominis) subtypes to other animals and human.

Dairy Farm Management Practices as Risk Factors Linked to Cryptosporidium spp. Infection in Dairy Calves

Background: Cryptosporidiosis is the disease of zoonotic importance and is considered to be third major cause of diarrhoeal disease of calves worldwide causing significant economic losses. The present study aimed to link Cryptosporidium spp. infections in dairy calves with the management practices as a major risk factor. Methods: Total 401 faecal samples from dairy calves were collected from eight large (100-1000 animals) and eight medium size (30-100 animals) dairy farms located in Jabalpur (Madhya Pradesh, India). Cryptosporidium oocysts were identified in faeces by modified Ziehl-Neelsen staining (mZN) and formal ether concentration techniques. Result: The prevalence of Cryptosporidium spp. was 28.7% in dairy calves. A higher prevalence and risk of cryptosporidiosis was associated with large farms (24.9% vs. 3.74%, OR=1.96), intensive farming system (27.7% vs. 0.99%, OR=1.86), drinking ground water (27.7% vs. 0.99%, OR=1.86), dirty farms (16.7% vs. 11.6%, OR=1.90), presence of other animals at farm (15.9% vs. 12.7%, OR=1.36), absence of deworming (14.7% vs.13.9%, OR=5.09) and diarrhoea treatment practice (16.2% vs. 12.5%, OR=1.07). Multi-animal calving area, dirty calving pen and cemented floor were also identified as risk factors. Practices like feeding colostrum within one hour of birth, @ 10% b.wt./day, weaning the calves within 24 hrs and using detergent to wash feeding utensils decreased the risk of infection from 1.4 to 2 times. Calves kept confined with their dam had greater prevalence and risk of infection (17.2% vs. 11.5%, OR=5.20). The findings highlight that proactive managemental practices are beneficial for decreasing the environmental contamination and improving calf health.

Using Constructed Wetlands to Remove Pathogenic Parasites and Fecal Coliforms from Wastewater in Dar es Salaam and Iringa, Tanzania

Wastewater treatment is a widely used health protection measure that can be applied to control the transmission of wastewater-related infectious diseases to communities exposed to wastewater. This study determined the efficiency of three full-scale constructed wetlands (CWs) in removing pathogenic parasites and fecal coliform (FC) bacteria from wastewater. Wastewater samples were collected from three CW systems located in the Dar es Salaam and Iringa regions of Tanzania. The modified Bailenger and modified Ziehl-Neelsen stain techniques were used to detect and quantify parasites. The membrane filtration method was used to detect and quantify FC bacteria. Data were analysed using IBM SPSS version 20. Helminth (Ascaris lumbricoides, hookworm, and Taenia spp.) eggs were completely removed by two CW systems. In all the systems, the removal of protozoa ranged from 99.8% to 100%. The mean concentrations of FCs in effluents ranged from 5 to 6 log units/100 mL. Effluents of all CW systems met the recommended parasitological quality requirements of the World Health Organization for the safe reuse of wastewater. FC effluents concentrations did not meet the local discharge standards of the Tanzania Bureau of Standards. Therefore, improvement to the CWs’ design, operation, and maintenance are required for the efficient removal of bacteria.
 Keywords: helminth, protozoa, fecal coliform, wastewater treatment, constructed wetland

Molecular characterization of algerian strains of Blastocysts sp

RésuméIntroductionBlastocystis sp. est un protozoaire colonisant le tube digestif de l’Homme et de nombreux animaux et il est à ce jour le parasite le plus fréquemment retrouvé dans les selles humaines. Dans certains pays en développement, sa prévalence dans les populations étudiées peut dépasser 50 %. Sur le plan morphologique, les isolats de Blastocystis sp. trouvés chez différents hôtes sont très similaires. Cependant, ces mêmes isolats présentent entre eux une très grande diversité génétique et pas moins de 17 sous-types (ou génotypes) ont déjà été identifiés à partir de données moléculaires. Des études de génotypage ont été réalisées dans de nombreux pays à travers le monde et en particulier dans certains pays du pourtour méditerranéen comme la France, l’Espagne, l’Italie, la Turquie ou l’Égypte. En revanche, très peu de données de génotypage sont disponibles en Algérie. À cet effet, nous avons réalisé la présente étude dont le but était d’identifier et de génotyper Blastocystis dans des échantillons de selles humaines et animales.Patients et méthodesMille huit cent soixante-neuf (1 869) prélèvements de selles appartenant à un personnel de cuisine dans le cadre du contrôle médical périodique, à des sujets pour la fourniture d’un certificat médical requis pour le traitement d’un dossier de visa et à des patients atteints de troubles gastro-intestinaux ont été examinés. À côté des selles humaines, des échantillons d’animaux, dont 10 volailles, 2 bovins et 2 murins ont été examinés. Toutes les selles ont été soumises à un examen microscopique direct complété par des techniques de concentration et une coloration de Ziehl Neelsen modifiée. La caractérisation moléculaire de 39 isolats humains et 14 d’origine animale a été réalisée par le séquençage et les séquences obtenues comparées à celles disponibles auprès de GenBank. Le séquençage n’a été contributif que pour 30 souches humaines et 9 souches animales.RésultatsSur l’ensemble des échantillons humain examinés, 284 étaient positifs (15,19 %) avec une prévalence de 7,38 % pour le Blastocystis. Parmi les 30 souches ayant fait l’objet d’une caractérisation moléculaire, le ST3 était prédominant (15/30, 50 %) suivi de ST1 (10/30, 33,33 %) et en troisième position le ST2 (4/30, 13,33 %). Le ST4 n’a été identifié que chez un seul patient (1/30, 3,33 %). La corrélation entre le statut clinique et le sous-type de Blastocystis identifié a montré que le nombre de ST3 était élevé chez les sujets asymptomatiques (11/15, 73 %) comparativement aux sujets symptomatiques (4/15, 26,66 %), de même pour le sous-type ST1 (7/10, 70 % versus 3/10, 30 %). À l’inverse, le nombre de ST2 était plus élevé chez les sujets présentant des troubles gastro-intestinaux (3/4, 75 %). À côté des souches humaines nous avons génotypé 7 souches aviaires, 2 souches murines et 2 souches bovines. La caractérisation des souches aviaires a révélé 5 ST6 (71,42 %) et 2 ST7 (28, 57 %). Les souches murines et bovines sont identifiées en tant que ST7 et ST6 respectivement.

Comparative diagnostic utility of metagenomic next-generation sequencing, GeneXpert, modified Ziehl-Neelsen staining, and culture using cerebrospinal fluid for tuberculous meningitis: Amulti-center, retrospective study in China.

BackgroundEarly diagnosis of tuberculosis meningitis (TBM) remains a great challenge during clinical practice. The diagnostic efficacies of cerebrospinal fluid (CSF)‐based mycobacterial growth indicator tube (MGIT) culture, modified Ziehl–Neelsen (ZN) staining, Xpert MTB/RIF, and metagenomic next‐generation sequencing (mNGS) for TBM remained elusive.MethodsA total of 216 adult patients with suspicious TBM were retrospectively enrolled in this multi‐cohort study. The diagnostic performances for MGIT, modified ZN staining, Xpert MTB/RIF, and mNGS using CSF samples were evaluated.ResultsUniform clinical case definition classified 88 (40.7%) out of 216 patients as the definite TBM, 5 (2.3%) patients as probable TBM cases, and 24 (11.1%) patients as possible TBM cases. The sensitivities of MGIT, modified ZN staining, Xpert MTB/RIF, and mNGS for TBM diagnosis against consensus uniform case definition for definite TBM were 25.0%, 76.1%, 73.9%, and 84.1%, respectively. Negative predictive values (NPVs) were 66.0%, 85.9%, 84.8%, and 90.1%, respectively. The sensitivities of MGIT, modified ZN staining, Xpert MTB/RIF, and mNGS for TBM diagnosis against consensus uniform case definition for definite, probable, and possible TBM were 18.8%, 57.3%, 55.5%, and 63.2%, respectively. Negative predictive values (NPVs) were 51.0%, 66.4%, 65.6%, and 69.7%, respectively. mNGS combined with modified ZN stain and Xpert could cover TBM cases against a composite microbiological reference standard, yielding 100% specificity and 100% NPV.ConclusionMetagenomic next‐generation sequencing detected TBM with higher sensitivity than Xpert, ZN staining and MGIT culture, but mNGS cannot be used as a rule‐out test. mNGS combined with Xpert or modified ZN staining could enhance the sensitivity of diagnostic tests for TBM.

Prevalence and Factors Associated with Cryptosporidium Infection in Calves in and around Nekemte Town, East Wollega Zone of Ethiopia.

Purpose Cryptosporidiosis, caused by a protozoan parasite of the genus Cryptosporidium, is a zoonotic disease that affects young animals and humans. The study was conducted from October 2020 to May 2021, with the objectives of estimating the prevalence of Cryptosporidium infection in calves and assessing the associated risk factors in Nekemte town, Ethiopia. Methods A cross-sectional study was adopted during the study period. Initially, 35 dairy farms were selected using a systematic study approach from which a total of 384 calves under the age of 12 months (75 calves from intensive, 51 from semi-intensive, and 258 from extensive dairy farms) were selected purposively for fecal sample collection. Fecal samples were collected directly from the rectum of each calf and tested for Cryptosporidium oocysts using Sheather's flotation and a modified Ziehl–Neelsen staining procedure. The data were analyzed using STATA statistical software version 13. Results Cryptosporidium infection was found in 53 of the 384 calves investigated, with an overall prevalence of 13.8% (95% CI = 8.1–17.6). Logistic regression analysis on the risk factors showed that types of farm, age, body condition scores, fecal consistency, types of feed, source of water, and farm hygiene were found to have a statistically significant influence on the shedding of Cryptosporidium oocysts by the calves (p < 0.05). Calves under the age of six months had a higher likelihood (OR = 2.7, 95% CI = 1.0–4.2, p < 0.001) of shedding the oocysts than the calves of 6 to 12 months of age. It was also confirmed that calves with poor body condition scores had a higher likelihood (OR = 2.6, 95% CI = 1.2–3.7, p=0.024) of shedding the Cryptosporidium oocysts than other ones. The likelihood of shedding a Cryptosporidium oocyst by the diarrheic calves was higher than by the nondiarrheic ones (OR = 3.1, p < 0.001). The infection was 2.6 times highest (16.8%, 31/185, 95% CI = 14.5–19.1, OR = 2.6, p=0.028) in calves feeding on grass alone, followed by milk and grass (15%) and milk (4.2%). Similarly, it was 2.2 times higher (18.5%, OR = 2.2, and p=0.002) in calves drinking river water than in calves drinking tap water. Moreover, calves kept under poor hygiene were more likely to shed Cryptosporidium oocysts (OR = 2.8, p=0.001) than calves kept under good hygiene. Conclusion In conclusion, cryptosporidiosis is one of the health problems of calves in and around Nekemte town, East Wollega zone. Our findings confirmed that risk factors such as types of farm, age, body condition scores, fecal consistency, types of feed, water source, and farm hygienic status were found to have a significant effect on the occurrence of Cryptosporidium infection in the calves. While the molecular-based study is required to be carried out aiming at species identification and the epidemiology of the parasite, particular attention should be paid to the control of the disease in the study area.

Subcutaneous Nocardiosis in a child with T- cell Immunodeficiency: A case report

Nocardia are weakly gram-positive, filamentous bacteria found worldwide in soils. Infection with Nocardia is rare inimmunocompetent patientsbut may leadto severe disease in immunocompromised patients. Reported cases in paediatric age group are few, and the literature is limited. We present a case of Nocardia isolated from pus from a skin lesion of eleven year old immunocompromised male child. Pus from Skin lesion on Ziehl-Neelsen (ZN) stain showed Acid Fast, Branching filamentous Bacteria. Pus culture on Lowenstein-Jensen (LJ) media showed glabrous, tough and waxycolonies. It was confirmed to be Nocardia species on Gram stain and modified ZN Stain. No organism was isolated on culture from Blood, urine and Cerebrospinal fluid (CSF). High clinical suspicion is required for diagnosis of slow growing bacteria such as Nocardia as they can be easily be missed on routine bacteriological culture.

Epidemiology of cryptosporidiosis in dairy calves in central and eastern Algeria

The purpose of our survey is to estimate the frequency of Cryptosporidium spp in dairy calves under 60 days of age, the pathogenic role of Cryptosporidium spp in the development of diarrhea, the intervention and association of other enteropathogens often involved in neonatal diarrhea and the possible relationship between infection and certain farming practices in order to estimate the share of risk factors, and on the other hand, to compare between the modified Ziehl-Neelsen staining method and the copro-ELISA test. We found 35 positive samples out of 223 faecal samples analyzed by modified Ziehl-Nielson staining, with a prevalence of 15.69%, the infected calves had 4 times the risk of diarrhea (RR= 4), the age group [8-15 days] is the most susceptible to infection compared to the other age groups (31.1%; p&lt;0.05). The ELISA test revealed that 33 out of 92 calves were infected by Cryptosporidium spp., with a prevalence of 35.87%, showing a sensitivity and specificity of Ziehl-Neelsen staining of 51.5% and 96% respectively.

Prevalence of Intestinal Parasitic Infection among Pre-School and School-Aged Children Attending to Fann University Hospital, Dakar: Results from Retrospective Analysis

Background: Intestinal parasitic infections represent a major public health problem worldwide. Pre-school and school children have a high risk of infection. The study aimed to determine the epidemiological aspects of intestinal parasitic infections in preschool and school-aged children in Dakar, Senegal. Methods: A retrospective analysis of laboratory records from preschool and school-aged children referred to the Laboratory of Parasitology at the Fann University Hospital in Dakar, Senegal, was carried out. The analysis of the period from 2016 to 2020. Stool samples were examined for the presence of parasites using direct, formal-ether concentration method and the Modified Ziehl Neelsen staining technique. Descriptive and analytic analysis was performed with Stata MP 16.1 software. Results: The overall prevalence of intestinal parasitic infection was 26.8% (429/1603). Blastocystis sp. (34%), Entamaeba coli (28%), Giardia intestinalis (9.6%), Endolimax nana (4.9%) and Ascaris lumbricoides (3.5%) were the common parasites found. Out of the 429 children positive for intestinal parasites, protozoa parasites represent 93.9%. Single infection and mixed infection represent 83.7% and 16.3%. The most prevalent associations were E. coli + Blastocystis sp. (6.3%), E. coli + E. nana (3.3%), and G. intestinalis + Blastocystis sp. (1.9%). Children between the ages of 5 - 10 years and those over 10 years had the most parasitic infections with 28.1% [aOR = 1.91; 95% CI (1.25 - 2.89)] and 27.4% [aOR = 11.92 (1.07 - 3.07)]. According to the year of sample collection, the prevalence was higher in 2017, 32.6% [aOR = 1.49 (1.11 - 2.01]. The prevalence of intestinal parasitic infection was not significantly related to gender (p = 0.87), season (p = 0.85) and symptomatology (p = 0.19). The main clinical symptoms were dysenteric syndrome, dyspeptic disorders, constipation and abdominal pain. Intestinal parasite carriage in patients with acute diarrhea was 20.4%. In HIV positive with chronic diarrhea, the prevalence was 28.6%. Conclusion: The results showed the frequency of intestinal parasites in preschool and school-aged children with a high proportion of protozoa parasites. Children aged over 5 years were more affected. Preventive measures such as health education, improved socio-economic conditions and environmental sanitation are required for reducing their frequency.

Epidemiology and public health implications of parasitic contamination of fruits, vegetables, and water in Kwara Central, Nigeria.

Fruits and vegetables are sources of parasitic infections in humans. This study was designed to determine the epidemiology of parasitic contamination of fruits, vegetables, and water, as well as the perception of retailers, buyers, and consumers about its contamination in Kwara Central, Nigeria. A cross-sectional study design was employed for this study. A total of 160 fruits, 349 vegetables, and 51 water samples were randomly purchased/collected from thirty different markets. The samples were subjected to sedimentation and modified Ziehl-Neelsen staining techniques. All the types of fruits and vegetables sampled were contaminated with one or more parasites. Eleven different species of zoonotic parasites (6 protozoans and 5 helminths) were detected. Entamoeba coli (39.8%) and Strongyloides stercoralis (9.6%) were the most prevalent protozoan and helminth respectively. The prevalence of other protozoan parasites ranged between 21.3% (Entamoeba histolytica) and 2.3% (Balantidium coli), while that of helminths was between 5.7% (Ascaris lumbricoides) and 2.1% (Trichuris trichiura). Source of fruits and vegetables, source of manure for cultivation, the source of water for washing/wetting purpose, market type, washing of fruits and vegetables before display, the presence of children at home and in the market, and knowledge of parasitic contamination were the risk factors associated (P<0.05) with parasitic contamination of fruits and vegetables among sellers. There is a need to educate the general public on the possibilities of parasitic contamination of fruits and vegetables and its public health implications.

Determination of microsporidia infection in larvae and adult anopheles mosquitoes (Diptera: Culicidae) Northwest Ethiopia

Microsporidia are a diverse group of obligate intracellular, spore-forming parasites that infect invertebrates including mosquitoes. A few studies showed that microsporidia infection of mosquitoes is associated with larval mortality or reduced adult fecundity and lifespan. This phenomenon might be used to protect against vector-borne disease transmission. This study aimed to determine microsporidia infection in field-collected Anopheles larvae and reared adults. Anopheles larvae were collected from the field and analyzed from February to April 2022. After determining the species identity of the 4th stage larvae, they were dissected to extract their midgut while the early-stage larvae were transported to an entomological laboratory for rearing the adult in the insectary room. Similarly, after determining the species identity of adult female Anopheles mosquitoes, they were dissected to extract their midgut. Then, two thin smears for each dissected larvae and/ adults were made. Fixed preparations were stained with Giemsa and modified Ziehl-Neelsen staining techniques to detect the microsporidia. Descriptive statistics and an independent t-test were used to analyze the data. Anopheles gambiae complex was identified as the most predominant species from field-collected larvae and insectary reared adults. From the total 258 larvae and 258 adults An.gambiae complexes tissue smears examined, microsporidia were detected in 2.7% [7/258, (95%, CI: 0.8-5)] and 1.2% [3/258, (95%, CI: 0.01-2.7)] of larvae and adults respectively. The mean density of microsporidia in larvae and adults was 64.9 (± 23.4 SD) and 36 (± 8.5SD) respectively. The difference in microsporidia density between infected larvae and adults was statistically significant (F=1.77, P= 0.02). Conclusion: Significant level of microsporidia infection was detected using light microscopy. Further microsporidia identification in the genus and species level is needed.

Prevalence and Risk Factors Associated with Intestinal Parasitosis in HIV/AIDS Patients on HAART in the Ndop Health District, North West Region, Cameroon

<i>Background:</i> Intestinal parasites induce an immunological alteration that favors progression from HIV to AIDS. These enteric parasitic agents vary from patient to patient and from country to country depending on endemicity, seasonal variation of pathogens, and also the immune status of the patient. Therefore, studies are required to evaluate the spectrum of these parasites in specific locations to better plan their control. The purpose of this study was to determine the prevalence and risk factors associated with intestinal parasitosis in HIV/AIDS patients on Highly Active Antiretroviral Therapy (HAART). <i>Methods: </i>A cross sectional study was carried out from the month of March to July 2018 on HIV patients on HAART in the Ndop Health District, Cameroon. Three centers were selected for this study using a two-stage stratified sampling technique. Stool and blood were collected from 347 participants. Stool was examined for intestinal parasites using direct microscopy, fomol-ether concentration technique, and modified Ziehl-Neelsen staining technique. Blood collected was analyzed using a Pima Alere CD4+ T cell counter. A structured questionnaire was administered to collect socio-demographic and clinical data. Data were analyzed with SPSS version 20 and p value < 0.05 was considered statistically significant. <i>Results:</i> Three hundred and forty-seven (347) consented individuals were recruited into the study. The ages of study participants ranged between 22-78 years. The mean (±SD) age of the study participants was 42 (±10) years. Among the 347 participants, 67 (19.3% (CI = 15.3%-23.9%)) were infected with at least one intestinal parasite. Forty-two (12.1% (CI =8.9%-15.9%)) of study participants were infected with protozoan while 28 (8.1% (CI= 5.2%-11.0%)) were infected with helminths. Intestinal parasites identified included; Ascaris<i> lumbricoides, Hookworm, Taenia spp, Schistosoma mansoni, Entamoeba histolytica/dispar, Cryptosproridum spp, Blastocystis hominis, Cyclospra spp, Isospora belli, Gardia intestinalis, and Entamoeba coli.</i> Low CD4+ T lymphocyte cell counts were significantly associated (P< 0.01) with intestinal parasitic infection in HIV/AIDS patients on HAART. <i>Conclusion: </i>Intestinal parasites are a problem to HIV/AID patient on HAART. Screening for intestinal parasites in HIV/AIDS patients on HAART must be highlighted and awareness created among HIV/AIDS patients.

Intestinal apicomplexan parasitoses among a hospital-based population in Honduras, 2013-2019.

: Introduction: Intestinal apicomplexa protozoa are a recognized cause of gastroenteritis. They are endemic in Honduras and their epidemiology varies in different population groups.Objective: To identify risk factors for cyclosporiasis, cryptosporidiosis, and cystoisosporiasis.Materials and methods: We conducted a case-control study in a hospital-based population. We performed the diagnosis using the modifed Ziehl-Neelsen staining technique and collected the information from laboratory records and clinical charts.Results: Cyclosporiasis was associated with diarrhea (OR=2.28; 95%CI: 1.10-4.89), weight loss (OR=12.7; 95%CI: 2.49-122.00), watery stools (OR=2.42; 95%CI: 1.26-4.65), and infection with another protozoan (OR=3.13; 95%CI: 1.66-5.95). Cryptosporidiosis was associated with HIV infection (OR=15.43; 95%CI: 3.34-71.22), diarrhea (OR=3.52; 95%CI: 1.40-9.40), lymphopenia (OR=6.16; 95%CI: 1.99-18.98), and green color stools (OR=3.00; 95%CI: 1.23-7.30). Cystoisosporiasis was associated with HIV infection (OR=11.20; 95%CI: 3.53-35.44), diarrhea (OR=7.30; 95%CI: 1.89-28.52), leukopenia (OR=4.28; 95%CI: 1.33-13.75), green color stools (OR=11.59; 95%CI: 1.16-558.60), and Charcot-Leyden crystals (OR=11.59; 95%CI: 1.16-558.60).Conclusions: In this hospital-based population from Honduras, HIV infection was a risk factor for cryptosporidiosis and cystoisosporiasis, but not for cyclosporiasis.

Genotyping of Cryptosporidium species in children suffering from diarrhea in Sharkyia Governorate, Egypt.

The protozoan parasite Cryptosporidium is one of the principal reasons for childhood diarrhea around the world. This work aimed to differentiate Cryptosporidium species among children suffering from diarrhea in Sharkyia Governorate, Egypt. A total of 97 fecal specimens were taken from children suffering from diarrhea, attending Pediatric Clinics of Zagazig University and Al-Ahrar Hospitals. Full history was taken. Stool samples were examined microscopically using modified Ziehl-Neelsen stain for detection of Cryptosporidium oocysts. To identify Cryptosporidium genotypes, positive samples were then subjected to nested Polymerase chain reaction-restriction fragment length polymorphism targeting Cryptosporidium oocyst wall protein gene. The overall detection rate was 27.8% (27/97) using modified Ziehl-Neelsen stain staining method. Using nested polymerase chain reaction, the gene was amplified in 85.2% (23/27). Restriction fragment length polymorphism analysis revealed that 65.2% (15/23) were Cryptosporidium hominis, 30.4% (7/23) were Cryptosporidium parvum, and one sample was not typed (4.4%). The significant risk factors associated with Cryptosporidium infection in children were animal contact and residence in rural areas. Cryptosporidium is a common enteric parasite affecting children in Sharkyia Governorate, Egypt, with the predominance of C. hominis genotype in children.

Investigation of the relationship between lymphocyte subsets and intestinal parasites

We analyzed the peripheral blood lymphocyte subsets of cancer patients infected with intestinal parasites, with an aim to find out the relationship between the levels of different types of lymphocytes with the prognosis of patients. 201 cancer patients aged 18 and over were included. Stool samples of the patients were examined using native-lugol, trichrome, modified trichrome (Weber's Trichrome stain), and modified Ziehl-Neelsen staining methods. Microsporidia and Cryptosporidium parvum were investigated at the genus and species levels using PCR. Lymphocyte subsets were determined by flow cytometry in blood samples. One or more parasite species were detected in 115 (56.7%) patients. The most common parasite species were Microsporidia, Blastocystis and Entamoeba coli, respectively. The frequency of parasites was high in patients with low lymphocyte percentage, CD3+ T cell and CD3+ CD4+ T (Th) cell levels in blood samples studied by flow cytometry. Microsporidia infection was significantly higher in patients with low lymphocyte percentage and Th cell levels. Similarly, C. parvum infection was found to be significantly higher in patients with low T lymphocyte percentage and Th cell level. Finally, Blastocystis infection was significantly higher in patients with low lymphocyte percentage and CD4/CD8 ratio higher than 1. The decrease in lymphocyte percentage, CD3+ T cell and Th cell count, and low CD4/CD8 ratio in cancer patients increase the frequency of intestinal parasitic infections. Based on these results, lymphocyte subsets may help identify cancer patients at high risk of opportunistic parasites. We suggest that opportunistic parasitic infections affecting the clinical course of the disease should be considered by clinicians during the follow-up and treatment of patients.

Prevalence of intestinal parasites in a cohort of HIVinfected patients from Antioquia, Colombia.

Introduction: HIV infection is still a public health problem worldwide and co-infections with other infectious agents including intestinal parasites are of particular concern, mainly in developing countries like Colombia.Objective: To conduct a cross-sectional study in patients attending an HIV care program in Antioquia given that there have been few intestinal parasites prevalence studies among the HIV population in the country.Material and methods: We evaluated stool samples from 192 patients by direct wet mount and concentration, modified Ziehl Neelsen staining, and agar plate culture. Univariate and correlation analyses were done to explore the association between socio-demographic and clinical characteristics and parasitological data.Results: The overall prevalence of intestinal parasites in HIV-positive subjects was 29.2% (56/192; 95% CI: 22.8% - 35.6%). Entamoeba histolytica/dispar/moshkosvkii with 13.0% (25/192; 95% CI: 8.2% - 17.8%) and Blastocystis with 12.0% (23/192; 95% CI: 7.4% -16.6%) were the most frequent. Opportunistic parasites like Cryptosporidium spp. and Cystoisospora belli were less prevalent, each one with 0.5% of positive samples (1/192; 95% CI: 0.1% - 1.5%). Commensal protozoa were also detected with a prevalence of 18.8% (36/192; 95% CI: 13.3% - 24.3%). Most of the individuals in the study had a controlled viral load and an LTCD4 count greater than 200 cel/µL. A small percentage (9.3%) had diarrhea. Bivariate analysis and multivariate logistic regression showed that only age and having pets had a significant association with intestinal parasites in this cohort.Conclusions: Our results confirmed that the evaluated population is at high risk of intestinal parasite infection, which highlights the need for routine screening of gastrointestinal parasites to provide prompt treatment and reduce possible complications.

A new single run polymerase chain reaction assay for cyclosporiasis in immunocompromised patients

causes human intestinal cyclosporiasis. It is more common in the immunocompromised patients and mainly seen in people living with HIV/AIDS (PLHA), post-renal transplant (PRT) patients and immunocompromised children (IC). Diagnostic microscopy for the oocysts of the parasite is less sensitive, requiring examination of multiple stool samples. Here we developed a new single run polymerase chain reaction (PCR) assay for the detection of and it was used to know the hospital based prevalence of cyclosporiasis. A cross-sectional study was conducted from June 2016 to October 2020 in a tertiary care teaching hospital. A new single run amplification PCR-based diagnostic assay was developed for . Stool samples were collected from 121 PLHA, 135 PRT and 79 immunocompromised children (IC) other than PLHA and PRT. All stool samples were examined for the presence of oocysts as well as tested with new PCR assay. Modified Ziehl-Neelsen staining of the concentrated stool smear did not reveal oocysts of species in any stool specimen. However, new PCR assay detected in 2 stool specimens – one from a PLHA patient and another from a PRT patient, giving a prevalence of 0.6% (2/335), 0.8% (1/121) in PLHA and 0.7% (1/135) in PRT. It was not detected in IC. Cyclosporiasis is infrequent in southern part of India. The new single run PCR assay developed by us is simple and cost effective molecular assay for the detection of .

PARASITIC CONTAMINATION OF TWO COMMONLY CONSUMED LEAFY VEGETABLES IN EL-KHARGA OASIS, UPPER EGYPT, AND EVALUATION OF HYGIENE PRACTICES AMONG THE VENDORS

Foodborne parasitic diseases cause human morbidity and mortality especially in the low- & middle-income countries. This study identified the parasitic contaminated Eruca sativa & Raphanussativus cultivated in El-Kharga Oasis, Upper Egypt, as to seasonal variation and the vendors hygienepractices. A total of 270 samples of the two vegetables were purchased from public markets.Parasitic contamination was evaluated by sedimentation and flotation techniques with modifiedZiehl-Neelsen stain. The results showed that 219(81%) samples were contaminated with 13different parasites. Ascaris lumbricoides 123(25.5%) and Cryptosporidium parvum (14.9%) werethe commonest ones and the highest contamination rate was in autumn (87%) for E. sativa and inwinter (87%) for R. sativus. The contamination level was significant with the gender and washingpractices of vendors. The current data gave evidence of the seasonal dynamics of acquiring parasiticinfection due to consumption of raw vegetables, interaction, and its relevance with vendor'shygiene.