Modified Ziehl-Neelsen Staining Research Articles

Rapid diagnostic tests relying on antigen detection from stool as an efficient point of care testing strategy for giardiasis and cryptosporidiosis? Evaluation of a new immunochromatographic duplex assay

Microscopy is the gold standard for the diagnosis of gastrointestinal parasites but is time-consuming and dependent on operator skills. Rapid diagnostic tests represent alternative methods but most evaluations have been conducted on a limited number of samples preventing their implementation in the clinical setting. We evaluated a new CE-IVD marked immunochromatographic assay (Crypto/Giardia K-SeT®, Coris Bioconcept) for the detection of G. intestinalis and Cryptosporidium spp. in 2 phases (retrospective and prospective) on a set of 482 stool samples including rare Cryptosporidium species. Besides G. intestinalis, this test could represent a rapid and reliable alternative to the modified Ziehl-Neelsen staining for the diagnosis of cryptosporidiosis (sensitivity/specificity were 89.2%/99.3% and 86.7%/100% for G. intestinalis and Cryptosporidium resp.), reducing diagnostic delays. Such strategy would also be time-saving by avoiding wet mount microscopy and concentrations steps, being particularly appropriate for laboratories having little expertise in microscopy or not able to implement molecular diagnostic methods.

Factors affecting the parasitic contamination of edible locally produced dry season leafy vegetables cultivated in south east Enugu, Nigeria

Background/Aim: Outbreaks of human infections associated with consumption of fresh fruits and vegetables have occurred with increased frequency during the past decade. This study evaluated parasitic contamination of locally produced dry season vegetables cultivated in Enugu metropolis, Enugu State, Nigeria. Materials/Method: Vegetables, water and soil samples selected from vegetable farms were investigated. A total of 160 vegetable samples, 15 soil samples and 15 water samples were investigated. The samples were washed separately in saline and the resulting solution was subjected to standard sedimentation and flotation techniques. Physiochemical analysis of soil and water samples was done using the standard DREM technique. Furthermore, the water samples were investigated for C. parvum using the modified Ziehl-Neelsen staining technique. Results: Parasitic contamination of the different vegetables were Teifeairia occidentalis 242 (18.1%), Amaranthus hybridus 193 (15.1%), Cucurbita maxima 192 (15.0%), Solanum species 191 (14.9%), Ocimum grattisimum 136 (10.6%), Solanun nigrum 114 (8.9%), Gongronema latifolium 101 (7.9%), Talinum triangulare 73 (5.7%) and Murraya koenigii 39 (3.0%).The parasites identified included; Ascaris lumbricoides , Taenia spp, Fasciola hepatica , Schistosoma spp, Trichuris spp, Enterobius vermicularis , Toxocara .spp, Giardia lamblia , Entamoeba spp and Cryptosporidium parvum . Entamoeba species was the highest contaminating parasite 529 (41.3%) while Schistosoma haematobium was the least,14 (1.1%). Physiochemical analysis revealed a strong positive correlation between parasite load in water and total suspended solutes (TSS) (r = 0.59, p = 0.021) and turbidity ( r = 0.55, p = 0.032) and also a strong positive correlation between parasite load in the soil and organic matter (OM), (r = 0.71, p = 0.003) and silt (r = 0.63, p = 0.011). Conclusion: These vegetables should be considered as potential sources of parasitic infections, especially when they are not properly cooked (the type called ‘Half Done’) or consumed raw like the local salad (abacha) because of the high risk of parasitic infections such as giardiasis, amoebiasis, enterobiasis, cyptosporidiosis, ascariasis, etc. Public enlightenment is necessary to enhance the adoption of effective food safety approaches by agronomic practices of vegetable farmers which will in turn reduce foodborne illnesses. Keywords: Parasite, contamination, vegetables, Enugu

Detection of Cryptosporidium spp. in Diarrheic Immunocompetent Patients in Beni-Suef, Egypt: Insight into Epidemiology and Diagnosis.

Cryptosporidium species is an important cause of gastrointestinal infections globally. This study aimed to shed light on its role in diarrheic immunocompetent patients in Beni-Suef, Egypt and to compare three diagnostic methods. Two hundred diarrheic patients, 37±16.8 year old, were enrolled. Stool samples were examined by light microscopy, using modified Ziehl-Neelsen stain (MZN) for Cryptosporidium spp. oocysts. Coproantigens were detected by sandwich ELISA. DNA molecular diagnosis was done by nested PCR. PCR yielded the highest detection rates (21.0%), compared to ELISA (12.5%) and MZN staining method (9.5%). The higher infection rates were in 20–40 year-old group, followed by 40–60 year-old. Association between epidemiologic factors was statistically not significant; positivity and gender, clinical manifestations, residence, source or water, or contact with animals. Cryptosporidiosis is an important enteric parasitic infection in Beni-Suef and PCR remains the gold standard for diagnosis.

Clinical and histological profile of leprosy patients at rural based tertiary care centre in post elimination era

Background: Despite India having achieved the national leprosy elimination goal of prevalence rate less than one case per 10000, the disease is still prevalent. Its diverse presentation depending on immune status of host warrants a clinico-histological correlation for diagnosis and start of proper therapy.Aim: The present study was done with the aim to describe the spectrum of clinical and histological profile of leprosy patients especially in doubtful and difficult cases especially where the clinical diagnosis is not sufficient and, then, to correlate clinical and histological profile.Methods: This study is a hospital based prospective study conducted on sixty newly diagnosed cases of leprosy. Skin biopsy was taken in each case and processed routinely. All the cases were stained with hematoxylin and eosin and modified Ziehl-Neelsen stain. Ridley-Jopling classification was used for diagnosis and typing of cases.Results: In the present study consisting of 60 newly diagnosed leprosy patients, majority of the cases i.e. 18 (30%) were in age group of 21-30 years with male: female ratio of 1.6:1. The most common type of skin lesion was plaque (50%), followed by hypo-pigmented macule (33.3%). The clinico-pathological concordance was maximum in Tuberculoid (TT) (100%) followed by Lepromatous (LL) (80%), Borderline lepromatous (BL) (72.7%), Borderline tuberculoid (BT) (50%) and Mid-borderline (BB) (33.3%).Conclusion: The overall clinicopathological concordance in present study is good (66.7%). Thus, the correlation of clinical and histological features along with bacillary index is far more useful for accurate typing and therapy in leprosy patients.DOI:10.21276/APALM.1680

Intestinal Parasitic Infestation in School Going Children of Rishikesh, Uttarakhand, India

Background: Intestinal parasitism(IP) remains a common problem in school going children. Aims & Objectives: To study the prevalence IP infestation in school going children and to study the relation with type of water supply and sanitation. Material and Methods: A cross sectional study was carried out and a total of 461 stool samples were collected from children under ten years of age from various schools of Rishikesh. The normal saline and iodine mounts of fresh and saturated salt solution concentrated samples were examined. Modified Ziehl Neelsen staining was done for detection of coccidian parasites. The statistical significance within gender, age, drinking water supply, sanitation and other socio-demographic parameters was assessed by Chi-square test. Results: 112 (24.3%) out of 461 were positive for intestinal parasites. Giardia lamblia was the commonest parasite (15.2%) followed by Hymenolepis nana (2.38%), Hookworm (2.17%), Enterobius vermicularis (2.17%), Entamoeba histolytica (2.17%), Ascaris lumbricoides (0.86%), and Cryptosporidium parvum (0.86%). Conclusions: Due to lack of proper sanitation & education along with low socioeconomic background, IP infections are still prevalent among school children. Regular awareness program on promoting good health, maintenance of proper personal hygiene and carrying out regular deworming in schools should be instituted for control.

Comparism of Various Staining Techniques in the Diagnosis of Coccidian Parasitosis in HIV Infection

Background: Diarrhoea due to oppourtunistic coccidian parasites is a common clinical presentation in HIV infection. Its management differs from that of diarrhoea due to other protozoa, improvement of immune status being the mainstay while specific drug treatment is available for other aetiologies, hence, the need for its accurate identification when present. This can be achieved via various diagnostic techniques, commonly microscopy in this environment, hence the need to compare the efficacy of the commonly used stains in our locality.
 Objective: To identify the most effective of the commonly used stains in identifying these parasites.
 Methods: Diarrhoea stool samples from 250 adult HIV positives and an equal number of age and sex matched HIV negative controls were screened, staining with trichrome, auramine and modified Ziehl–Neelsen stain.
 Results: A positivity rate of 55% was reported. modified Ziehl–Neelsen , when compared with trichrome staining had 81% sensitivity, 77.3% specificity, positive predictive value of 70.4%, negative predictive value of 85.9% and when compared to auramine staining, had 80% sensitivity, 76.7% specificity, positive predictive value of 69.9%, negative predictive value of 85.2% in test subjects. There was a significant moderate level of agreement between the staining methods though Trichrome showed a stronger agreement than Auramine when compared with Modified ZN in test (κ value 0.569 and 0.553 respectively), and a significant, fair level of agreement between the methods with Auramine showing a stronger agreement than Trichrome when both were compared with Modified ZN (κ value 0.399 and 0.332 respectively) in controls.
 Conclusion: Auramine and trichrome techniques are preferred for screening and diagnosis based on findings. Of these two techniques, auramine is preferred.

Prevalence of Opportunistic Intestinal Parasites and Associated Factors among HIV Patients while Receiving ART at Arba Minch Hospital in Southern Ethiopia: A Cross-sectional Study.

Opportunistic Intestinal Parasites (OIPs) commonly cause morbidity in HIV/AIDS patients due to the decline of CD4+T-cells. The burden of Opportunistic Intestinal Parasitic Infections (OIPIs) in Ethiopia is expected to be high due to poor sanitation and co-pandemicity of HIV/AIDs. Therefore, frequent assessment of the magnitude and associated factors for intestinal parasitosis is essential for the management of HIV/AIDS patients. A hospital based cross-sectional study was conducted among patients attending Arba Minch Hospital Antiretroviral Therapy (ART) Clinic from March to June 2016. Stool specimens were processed for parasitological examination using direct wet mount, formol-ether sedimentation and modified Ziehl-Neelsen staining techniques. CD4+ T-cell count data were taken from patients' medical records. A structured questionnaire was used to collect data on socio-demographic characteristics and possible associated factors for OIPIs. All the data were analyzed using SPSS version 20. Two hundred and twenty ART patients participated in the study. The overall prevalence of intestinal parasitic infections was 28.18% while that of OIPIs alone was 17.72%. Among identified intestinal parasites, Cryptosporidium species accounts for the highest frequency (19/220, 8.63%), followed by Cyclospora species (13/220, 5.90%). Presence of domestic animals (AOR=2.07, 95%CI:1.07-8.40, P= 0.032) and CD4+ T-cell count <500cell/µl (AOR=4.66, 95%CI:1.17-5.35, P= 0.001) were significantly associated with OIPIs. The study indicated that co-infection rate of OIPs is high among ART patients. It also found that contact with domestic animals and having CD4+ count <500 cell/µl predict for the presence of OIPs.

Molecular epidemiology of Cryptosporidium spp. in North Lebanon.

Cryptosporidium spp. are enteroparasites with worldwide distribution that infect the gastrointestinal tract of several vertebrates including humans. Human to human, zoonotic, foodborne and waterborne are reported as the main transmission routes of this parasite. Cryptosporidium spp. have been recognized as the predominant cause of waterborne and foodborne outbreaks. However, the epidemiological situation of cryptosporidiosis is not well known in Lebanon, a developing country with a population often affected by intestinal parasitic infections. This study was devoted to determine the prevalence and the genetic diversity of Cryptosporidium spp. in symptomatic hospitalized patients and in two children populations with different socio-economic level in North-Lebanon, as well as the risk factors associated with cryptosporidiosis. Fecal samples obtained from these populations were examined microscopically by modified Ziehl-Neelsen staining as well as nested PCR were done for the detection of Cryptosporidium oocysts. Out of 163 symptomatic hospitalized patients and 249 children, Cryptosporidium was present in 11% and 10.4% respectively according to microscopy examination and/or molecular tests. The genotyping showed the predominance of Cryptosporidium hominis in both populations. Subgenotype analysis of the isolates at the gp60 locus identified three subtypes IdA19, IbA10G2 and IaA18R3 for C. hominis and two subtypes IIaA15G1R1 and IIaA15G2R1 for C. parvum. Moreover, cryptosporidiosis was correlated with having meals outside home and presence of gastrointestinal symptoms especially diarrhea (p <0.05). This work constitutes the first molecular epidemiology study outlining risk factors associated with cryptosporidiosis in Lebanon. These findings support a need of a control program to prevent the circulation of this parasite.

Submandibular and parotid abscess due to Nocardia sp. and therapy with danofloxacin in sheep

The aim of the present study was to report detection of Nocardia sp. from cutaneous lesions of sheep. Abscess material was collected from submandibular and parotid lesions in sheep for identification of Nocardia sp., which is well known as a causative agent for cutaneous or subcutaneous abscesses, disseminated disease, mastitis, and pneumonia. Nocardia sp. are Gram-positive, delicate, non-motile, branching filamentous bacteria that found in soil, dust, organic materials, water and aquatic environmental. Nocardia species generally yield a large variety of diseases in both normal and immunocompromised humans and animals. In the present study, bacterial staining methods and biochemical identification methods were applied for identification of Nocardia sp. in four sheep in Aydin. All cases had superficial abscesses in the submandibular and parotid region, mostly invading the head and neck regions. Neutrophilic leukocytosis was evident in three out of four sheep, and there was evidence of moderate anemia in two of the sheep. Samples were stained with Gram and modified Ziehl-Neelsen staining and streaked onto sheep blood agar, incubated at 37°C for 48-72 hours under aerobic condition, Sabouraud Dextrose Agar, incubated for one week under aerobic condition. Nocardia sp. was isolated from based on bacteriological investigation confirming numerous thin, Gram-positive, filamentous and nonacid fast organisms, and biochemical tests. Nocardia sp. strains were confirmed with 16s Rrna PCR method. Based on clinical appearance, hematological findings, and molecular analysis a definitive diagnosis cutaneousNocardiosis was made in the sheep. Initial therapy involving danofloxacin, with regard to antibiogram, for 10 days and local iodine solutions, resulted in clinical recovery on 35-45 days post-treatment. This is the first reported case of cutaneous Nocardiosis and its effective danofloxacin therapy in sheep in Turkey.

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..

Evaluation of mycobacteria-specific gamma interferon and antibody responses before and after a single intradermal skin test in cattle naturally exposed to M. avium subsp. paratuberculosis and experimentally infected with M. bovis

This study reports on the diagnostic potential of IFN-γ release assays and serology for Mycobacterium bovis in six naturally M. avium subsp. paratuberculosis (Map) exposed bulls of which four were intratracheally infected with a Belgian field strain of M. bovis. Heparinized blood, serum and fecal samples were collected at regular time intervals for mycobacteria-specific IFN-γ release assays, antibody analysis and for Map culture respectively. Single intradermal skin test (SIT) with bovine tuberculin (PPD-B) was performed on day 115 and animals were sacrificed on day 133 after M. bovis infection. Organs were collected and stored for histopathological examination, modified Ziehl-Neelsen staining and bacteriological analysis of M. bovis and Map by culture and RT-PCR.Prior to infection five animals showed positive IFN-γ responses to avian PPD (PPD-A) and four were positive in Map PCR (IS900) on faeces. Three M. bovis infected animals reacted as early as day 14 with sustained higher PPD-B than PPD-A specific IFN-γ responses, whereas the fourth animal (with the strongest PPD-A response prior to infection) showed sustained higher PPD-B specific IFN-γ levels only a day 56 after infection. Two of the infected animals had a sustained positive IFN-γ response to the ESAT-6/CFP-10/TB7.7 (QuantiFERON®-TB Gold) peptide cocktail as early as day 14, among which the animal with the initial high PPD-A response. Later during infection, positive responses were found to ESAT-6 peptides in three infected bulls and to CFP-10 peptides in all four infected bulls. One of the control animals reacted intermittently to the ESAT-6/CFP10/TB7.7 cocktail. Prior to SIT, weak but positive MPB83/MBP70 specific antibody responses were detected in two of the infected bulls. All four M. bovis infected bulls reacted with a positive skin test and showed, as reported by others, increased mycobacteria specific IFN-γ production and increased positive responses in MPB83/MBP70 specific serology after SIT. At autopsy, M. bovis lesions were detected in all four experimentally infected bulls.Our results indicate that in Map exposed cattle, M. bovis diagnosis using IFN-γ assays needs a combination of PPD-B/A and ESAT-6/CFP10 for early and optimal sensitivity and that sensitivity of MPB83/MBP70 serodiagnosis is dramatically increased by prior skin testing. Map exposure did not interfere with the development of SIT in M. bovis infected animals, but resulted in a false positive M. bovis specific IFN-γ and antibody response after SIT in one of the two control animals (which remained negative in skin-test).

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.

Detection rates of waterborne protozoa in water sources from Fayoum Governorate

Background: Waterborne protozoal infections are common health problems in many parts of the world especially in developing countries. Water is a major vehicle for transmission of protozoa such as G. lamblia, Cryptosporidium spp. as well as pathogenic and opportunistic free living amoeba (FLA).Objective: This study aims to detect the presence of protozoal agents in tap water and storage water tanks at Fayoum Governorate, Egypt.Material and Methods: A total of ninety five water samples were collected from different water sources, taps (65) and tanks (30), from 6 Fayoum districts. The samples were processed to detect the presence of G. lamblia cysts by Lugol’s iodine stain, Cryptosporidium oocysts by modified Ziehl-Neelsen stain and FLA by cultivation. After cultivation, Acanthamoeba spp. were identified according to their morphological features and flagellation test was performed to detect amoeboflagellates.Results: All water samples collected from tanks (100%) were contaminated by protozoa of medical importance, while only 6 (9.2%) of the tap water samples were pathogen free. The majority of water samples were contaminated with mixed protozoal infections. The overall detection rates of contaminants in water sources were 86.3%, 52.6%, 13.7% by FLA, Cryptosporidium spp. and G. lamblia, respectively.Conclusions: The recorded detection rates of waterborne protozoa present a hazard to the community resulting in silent morbidities and mortalities. It is strongly recommended to adopt proper water safety measures.

Intestinal parasitic infection in adult patients attending tertiary care hospitals: a retrospective study

Background: Intestinal parasitic infections (IPI) are recognized as neglected tropical diseases. Inadequacy of epidemiological data reporting the prevalence of intestinal parasites among the general population prompted this retrospective study conducted among the Departments of Medicine and Microbiology of two medical colleges.Methods: All the symptomatic patients were examined for IPI. 831 parasite positive stool samples were included for the study. The age range of the patients were above 18 years. Stool specimens were processed using formol-ether concentration technique. Routine microscopic examination (saline and Lugol’s iodine mounts) were carried out for ova, cysts or parasites. The sample was partly preserved in 10% formal saline and later concentrated and fixed with methanol before staining by modified Ziehl Neelsen stain. Results: Out of the 831 positive samples, E. histolytica was predominant with 337 (40.55%) of all cases. The next in order were Giarrdia lamblia 216 (25.99%), A. lumbricoides with 173 (20.81%), E. vermicularis with 43 (5.17%), A. duodenale with 23 (2.76%), H. nana with 19 (2.28%), S. stercoralis with 17 (2.04%) and Taenia with 3 (0.36%). 134 patients had double parasites isolated, while only 3 had triple parasitic infestation.Conclusion: Present study showed a preponderance in gender towards males (59.4%) than females (40.5%). Larger sample size and more surveillance studies will help to establish effective measures and evidence-based approaches to curb the parasitic proliferation in this region.

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.

COMPARISON BETWEEN AURAMINE PHENOL STAIN, MODEFIED ZIEHIL-NEELSEN AND ELISA FOR DETECTION OF INTESTINAL COCCIDI

Coccidia had been implicated as the most important opportunistic parasites in patients with acquired immune deficiency syndrome. It is transmitted via the feco-oral route. The study aimed to compare the efficacy of microscopic tests with ELISA assay for detection of oocysts in feces. This study assisted the presence of Coccidia oocyst in fecal samples of 300 patients attending Zagazig University Hospital by using modified Ziehl-Neelsen stain (MZN) ,Auramine phenol stain (AP) and Enzyme-linked immunosorbent assay. Cryptosporidium infection rate was the highest (20.7%) among other Coccidia. Mixed infection of Cryptosporidium and Cyclospora either alone or with Cytoisospora and that of Cryptosporidium and Cyclospora with concomitant Microsporidia were recorded (9.7%, 1.7% and 5% respectively) by AP and (17.7, 6.7, 1 and 3.3 % respectively) by MZN. The specificity of modified Ziehl-Neelsen was 100% for both, while the sensitivity was 77.5%, while they were 100% and 100% respectively for AP, and 100% and 91.9% for ELISA. Conclusion, this study shows that Auramine phenol is a simple fluorescent staining, promising technique in diagnosis of intestinal Coccidia as it has high sensitivity and specificity and it is less time-consuming method.

Prevalence of Cryptosporidium Infection among HIV-1 Infected Adult Patients Attending Jos University Teaching Hospital, Jos, Nigeria

Cryptosporidium is a leading cause of chronic diarrhoea in HIV-1infected patients, but there is paucity of data on the prevalence of Cryptosporidium and factors associated with the infection. We determined the prevalence and factors associated with Cryptosporidium infection among HIV-1 infected adult patients at Jos University Teaching hospital, Jos, Nigeria. A cross-sectional study in which a total of 296 fecal specimens from HIV-1 infected patients with diarrhea was collected and structured questionnaire was used to collect demographic and clinical data. The fecal samples were examined for Cryptosporidium by formol-ether concentration and modified Ziehl-Neelsen staining technique, and the oocysts identified by microscopy. Two hundered and ninrty six feacal samples where analyse from156 (52.7%) females and 140(47.3%) males. The prevalence of Cryptosporidium infection among HIV-1 infected patients was 4.7%. Among females 9 (5.8%) had Cryptosporidium infection while (5)3.6% in males; P=0.696. The age group 21-30 years had the highest prevalence of (8)10.0%. A statistically significant association of Cryptosporidium infection was observed with rural type of residence, P=0.046. In conclusion the prevalence of Cryptosporidium infection among HIV-1 infected adult patients with diarrhea was low. This may be due to early commencement of antiretroviral drugs and the use of anti-parasitic prophylaxis among the patients. Further larger studies on the molecular identification of the species of Cryptosporidium are needed to determine the available zoonotic strains.

Comparison of various staining techniques in the diagnosis of Coccidian parasitosis in HIV infection

Diarrhoea due to oppourtunistic coccidian parasites is a common clinical presentation in HIV infection. Its management differs from that of diarrhoea due to other protozoa, improvement of immune status being the mainstay while specific drug treatment is available for other aetiologies, hence, the need for its accurate identification when present. This can be achieved via various diagnostic techniques, commonly microscopy in this environment, hence the need to compare the efficacy of the commonly used stains in our locality. The objective of the study is to identify the most effective of the commonly used stains in identifying these parasites. Diarrhea stool samples from 250 adult HIV positives and an equal number of age and sex matched HIV negative controls were screened, staining with trichrome, auramine and modified Ziehl Neelsen stain. A positivity rate of 55% was reported. Modified Ziehl Neelsen, when compared with trichrome staining had 81% sensitivity, 77.3% specificity, positive predictive value of 70.4%, negative predictive value of 85.9% and when compared to auramine staining, had 80% sensitivity, 76.7% specificity, positive predictive value of 69.9%, negative predictive value of 85.2% in test subjects. There was a significant moderate level of agreement between the staining methods though trichrome showed a stronger agreement than auramine when compared with Modified ZN in test (κ value 0.569 and 0.553 respectively), and a significant, fair level of agreement between the methods with Auramine showing a stronger agreement than trichrome when both were compared with Modified ZN (κ value 0.399 and 0.332 respectively) in controls. Auramine and trichrome techniques are preferred for screening and diagnosis based on findings. Of these two techniques, auramine is preferred.Journal of Medical and Biomedical Sciences (2016) 5(3), 28-35Keywords: comparison, trichrome, auramine, modified ziehl neelsen, HIV

Dogs' Gastrointestinal Parasites and their Association with Public Health in Iran.

IntroductionDogs harbour zoonotic parasites that cause serious infections in humans, such as visceral larva migrans, ocular larva migrans, cystic echinococcosis, and alveolar echinococcosis. Studies on dogs’ gastrointestinal parasites in different geographical locations are required to increase knowledge of the risk of canine zoonoses in human populations.Material and MethodsThe presence of parasites was examined in 450 faecal samples collected from eight zones of Zanjan province, northwest Iran from June to November 2015. The samples were examined using the sedimentation concentration method and modified Ziehl-Neelsen staining.ResultsGastrointestinal parasites were found in 86 (19.1%) faecal samples. Sarcocystis spp. (7.3%), Taenia/Echinococcus spp. (5.6%), Toxocara spp. (1.8%), and Cystoisospora spp. (1.6%) were the most common parasites observed. The other detected parasites consisted of Dicrocoelium dendriticum (0.7%), Eimeria spp. (0.7%), Cryptosporidium spp. (0.4%), Physaloptera spp. (0.4%), Giardia spp. (1.3%), and Spirocerca lupi (1.3%). The lowest parasite infection rates belonged to Trichuris vulpis and Acanthocephalans (0.2% each).ConclusionThis study provides current information on the infection rates in dog populations in Zanjan Province. Furthermore, the study shows a high prevalence of gastrointestinal parasitic infections, including zoonotic ones and particularly Taenia/Echinococcus spp., potentially transmissible to humans and thus relevant to public health.