Modified Ziehl-Neelsen Staining Research Articles

Molecular genetic characterization of human Cryptosporidium isolates and their respective demographic, environmental and clinical manifestations in Egyptian diarrheic patients

Background: Cryptosporidium is a zoonotic protozoan parasite of public health and veterinary importance, causing gastroenteritis in a variety of vertebrate hosts. Several Cryptosporidium spp. with different genotypes and subtypes are implicated in human cryptosporidiosis. Objective: Detection and correlation of native Cryptosporidium molecular genetic variability among Egyptian diarrheic human isolates with their respective demographics, environmental and clinical manifestations. Patients and Methods: A total of 350 human stool samples were collected from diarrheic patients. The collected samples were examined microscopically and by RIDA®QUICK Cryptosporidium/Giardia copro-immunochromatography test (ICT) cassettes and strips. All positive Cryptosporidium samples detected by modified Ziehl-Neelsen stain (MZN) and/or ICT were genotyped using nested polymerase chain reaction-restriction fragment length polymorphism (nPCR-RFLP) targeting cowp gene encoding Cryptosporidium oocyst wall protein ( Cowp). Results: Cryptosporidium was detected in 5.7% (20/350) of the examined samples using MZN, 9.4% (33/350) by ICT kit, and 3.7% (13/350) by both methods. A total of 40 samples were positive by both microscopy and/or ICT. Only 15/40 (37.5%) of samples positive by microscopy and/or ICT were successfully amplified using cowp gene. All amplified samples (15) were confirmed as C. hominis by nPCR-RFLP analysis of the cowp gene. Significant associations were found between cryptosporidiosis and gender, drinking groundwater, immunodeficiency of patients, complaining of fever, watery and mucoid stool, consumption of readymade salad, and unwashed fresh vegetables. Conclusion: In the current examined samples, C. hominis proved to be the dominant genotype, indicating that transmission of cryptosporidiosis among a sample of diarrheic Egyptian patients is most probably anthroponotic rather than zoonotic. MZN and PCR gave a diagnostic accuracy of 100%, while ICT diagnostic accuracy proved to be 50%.

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.

A Study on Intestinal Parasitic Infections Among Immunocompetent Egyptian Children Attending Al-Hussein University Hospital, Cairo

Background: Parasitic infections are still a dangerous public health problem in the world, especially in developing countries including Egypt representing a major cause of morbidity and mortality mainly among children. Methods: This cross-sectional study included 400 randomly chosen immunocompetent children of both sexes. Their ages ranged from 6 months up to 12 years old. All children were subjected to a structured questionnaire sheet, stool examination using: Direct smear and concentration techniques (formol ether sedimentation and Modified Ziehl–Neelsen stain). The study was approved by the medical ethics committee of Al-Azhar University Hospitals and a written informed consent was obtained from all children's parents or gardeners. Results: Among the examined individuals, parasites were detected in only 139 ones (34.75%) mainly in the first two years of age and more common among males 35% than females 34%. By species, the parasitic prevalence rate was, G.lamblia 49 (12.25%), E.vermicularis 35 (8.75%), E. coli 22 (5.5%), E. histolytica/dispar 7 (1.75%), H. nana 4 (1.0%) and mixed infections 22 (5.5%). Positive results are more common among rural children 38% and low social classes. Low level of parent's education, low family income, high crowding index, and bad hand hygiene were considered highrisk factors of parasitic infections. Conclusion: Rural residency, bad hygiene, and low social class, as well as crowdedness, were the most significant risk factors associated with parasitic infections among children

Identifikasi Protozoa Usus Oportunistik dan Faktor Risikonya Pada Anak Panti Asuhan Kota Pekanbaru

Opportunistic intestinal protozoa are protozoa that can infect humans at a time when the body’s defense system isdeclining (immunocompromising). Opportunistic protozoan infections are infections by intestinal protozoa that werenot previously considered important, but now can cause disease in humans. Opportunistic intestinal protozoan infectionsbesides being found in immunocompromised patients are also reported to infect children. Some of the intestinalprotozoan species that have been identified to cause infection are Cryptosporidium sp, Isospora belii, Cyclospora sp,and Blastocystis hominis. This study was an analytical study with a cross sectional design conducted in June 2018until January 2019. In this study, modified Ziehl-Neelsen staining was used and the results were 25.6% of orphanagesinfected with opportunistic intestinal protozoa, which consisted of Cryptosporidium sp. as much as 14.1%, Blastocystishominis as much as 4.6%, infection with a mixture of Cryptosporidium sp and Blastocystis hominis as much as 4.6%,Isospora belii as much as 2.3%, and no infection by Cyclospora cayetanensis. There was no association betweenopportunistic intestinal protozoan infections with poor handwashing habits, bowel habits, food hygiene, drinkingwater treatment, and bad animal raising habits.

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.

Prevalence of Intestinal Parasitic Infections in Relation to the HIV Status of Patients Attending the Care Units in Three Divisions in the Centre Region of Cameroon

Background: Depression of the immune system caused by the human immunodeficiency virus (HIV) promotes the onset of opportunistic infections including intestinal parasites. Objectives: In this study, we are comparing the prevalence of these parasitic infections among individuals infected with HIV (HIV+) and non-infected (HIV-) enrolled in four HIV care units in the Centre Region of Cameroon. Methods: A cross-sectional study was conducted and stool samples were collected from 283 HIV positive subjects and 245 seronegative subjects. These samples were processed using direct wet mount, formol-ether concentration, Kato-Katz procedure, modified Ziehl-Neelsen staining, and Baermann techniques to identify both common and opportunistic intestinal parasites. Logistic regression analyses were used to assess the association between HIV infection and socio-demographics factors as well as infection with intestinal parasites. Results: A total of 123 (23.4%) individuals were found infected with at least one protozoan parasite species, and nine (1.7%) with intestinal worms. Overall, infection rates were 27.9% and 22.4% among HIV+ and HIV- subjects, respectively. The frequencies of Cryptosporidium spp. and Entamoeba histolytica/dispar were significantly more important among HIV+ subjects (P < 0.0455). Pentatrichomonas hominis and Entamoeba coli were significantly more prevalent among HIV- subjects (P < 0.0210). Heterogeneity was observed in the distribution of intestinal parasites, according to socio-demographic parameters. In addition, Cryptosporidium spp. was significantly associated with the decrease in CD4 cell count (P = 0.0035). The frequencies of infections with Cryptosporidium spp., Isospora belli, and Iodamoeba buetschlii were significantly more important in HIV positive patients not taking antiretroviral (P < 0.0226). Conclusions: Cryptosporidium spp., Entamoeba histolytica/dispar, and Blastocystis spp. were the intestinal pathogens more often found among HIV infected individuals. HIV positive subjects were significantly more infected with intestinal parasites than seronegatives in intermediate and rural settings, among individuals aged more than 45 years and among unemployed individuals.

Cutaneous nocardiosis with discharging sinus clinically mimicking tuberculosis diagnosed by cytology.

Nocardiosis is primarily a pulmonary infection commonly seen in immunocompromised individuals. However, lymphocutaneous nocardiosis is observed in immunocompetent individuals often after trauma. The clinical and cytomorphological features of lymphocutaneous nocardiosis closely mimic the most common infections in India such as tuberculosis and mycetoma (very common cutaneous infection with discharging sinus). As it is crucial to differentiate nocardiosis from tuberculosis, to avoid unnecessary antitubercular treatment, special stains like modified Ziehl-Neelsen stain and Gram stain can be employed to differentiate the morphology of Nocardia from tuberculosis. Fine-needle cytology from these cutaneous lesions helps in yielding adequate material for rapid and accurate diagnosis of immediate specific antibiotic treatment. We report a rare case that presented with clinical diagnosis of tuberculosis but turned out to be nocardiosis on cytomorphology with simple and most feasible fine-needle aspiration method of tissue diagnosis and scrape cytology.

Intestinal Parasites Associated with Opportunistic Coccidial Infections Among Immunocompromised Individuals in Central Iran: A Cross Sectional Study

Background: Protozoa and helminthic parasites are the most common opportunistic parasites infections associated with the gastrointestinal tract in immunocompromised patients. Objectives: There have been very few studies addressing this issue in central Iran and our purpose was to determine the frequency of the intestinal parasitic infections (IPIs) in different groups of immunocompromised patients admitted to the referral hospitals in Isfahan, Iran. Methods: A cross-sectional study was performed on 204 immunocompromised patients (HIV/AIDS, lymphoma, leukemia, renal transplant and other transplants) between 2015 - 2016. Stool samples were analyzed for intestinal parasites using direct-smear, formol-ether concentration method and modified Ziehl-Neelsen staining techniques. Results: The total rate of any parasites was 43.1% (88/204) in the patients. The prevalence of parasites was 32.7% (17/52), 39.6% (19/48), 46.2% (18/39), 56.0% (28/50), and 40.0% (6/15) in HIV/AIDS, lymphoma, leukemia, renal transplant recipients, and the other transplant recipients, respectively. Blastocystis hominis (30.4%), Cryptosporidium spp. (3.9%), Entamoeba coli (6.3%), Giardia lamblia cyst (5.4%), Endolimax nana (2%), ova of Fasciola spp. (0.5%) and Dicrocoelium dendriticum (0.9%) were the overall parasites that were found in this study. The most common parasites which were related to diarrhea were Blastocystis hominis and Cryptosporidium spp. The parasitic infection was significantly higher in urban patients and females (P < 0.05). Nevertheless, no significant relationship was observed between the prevalence of parasitic infections and age, occupation and level of education. Conclusions: Our findings highlighted that IPIs are a common health problem among immunocompromised patients, in central Iran. Therefore these patients should be screened routinely for intestinal parasites and treated promptly.

Prevalence and risk factors of intestinal parasite infections in Mazandaran province, North of Iran

BackgroundIntestinal parasitic infections (IPIs) are confirmed as major public health problems, worldwide. Northern Iran is highly endemic area for such infections. This research aimed to study on the prevalence of IPIs among people referred to the health care centers in Mazandaran Province, Northern Iran. MethodsIn this descriptive cross-sectional survey, 984 stool samples were collected during 2015–2016. Fresh stool samples were collected from all individuals and examined by direct slide smear, formalin-ether concentration, trichrome and modified Ziehl-Neelsen staining techniques. The results were analyzed using SPSS version 16 and Chi-square test. ResultsOut of 984 participants, the overall prevalence of IPIs was 12.1% (95 CI, 11.4–12.7; 119/984). The prevalence of protozoa and helminthes infections were 7.9% (95 CI, 7.4–8.3; 109/984) and 5.3% (95 CI, 4.9%–5.5%; 69/984), respectively. The most prevalent protozoan IPs were include Giardia lamblia (3.3%), Entamoeba coli (2%), Blastocystis hominis (1.3%), Entamoeba histolytica/dispar (1.1%), Entamoeba hartmanni (1.1%) and Cryptosporidium spp. (0.6%). Also major helminthic infections were include Trichostrongylus spp. (2.6%), Hookworms (1.7%), Strongyloides stercoralis (1.1%), Hymenolepis nana (0.6%) and Dicrocoelium dendriticum (0.6%). The results of the univariate analysis revealed that residence (rural being [P = 0.017]) and occupation (farmers [0.033]) were statistically significant risk factor for IPIs. ConclusionIn conclusion, we have found, despite persistent improvement in sanitary conditions over the recent years in northern part of Iran, IPIs are still among the main public health problems in this area. Moreover, helminths infection in our study were considerable. Thus, effective control programs are essential to reduce the incidence of IPIs.

Causative agents for diarrhoea in under 5 children in a tertiary care hospital

Background: nfectious diarrhoea is a major public health problem in developing countries like Bangladesh. Bacteria and parasites are important etiological agents for diarrhoea in infants and children.&#x0D; Objectives: To isolate and identify the causative organisms of diarrhoea from diarrhoeal stools in under 5 children.&#x0D; Methodology: This was an observational cross sectional study carried out at Microbiology Department of Sir Salimullah Medical College, Dhaka. A total of 217 diarrhoeal patients upto 5 years age, attending Pediatric unit of Sir Salimullah Medical College and Mitford Hospital, Dhaka and Dhaka Shishu Hospital, Dhaka during the period of July 2015 to June 2016, were enrolled in the study. For isolation of enteric bacteria, diarrhoeal stools were inoculated on Mackonkey agar media and for Campylobacter spp. diarrhoeal stools were inoculated on selective Modified Campy-BAP media and nonselective Blood agar media with membrane filter. Positive culture was confirmed by Gram stain morphology and standard biochemical tests. Oocysts of Cryptosporidium were identified by Modified Ziehl Neelsen staining and other parasites were identified by wet mount preparation.&#x0D; Results: Out of 217 diarrhoeal stools bacteria were isolated from 97 (44.7%) patients. Among 97 cases, Escherichia coli were detected in 51(52.58%) patients; Campylobacter spp. were detected in 23(23.71%) patients; Shigella spp. were detected in 15(15.46%) patients and Salmonella spp. were detected in 8(8.25%) patients. Microscopic finding of diarrhoeal stools were detected by pus cells with or without RBCs in 90(70.31%) samples. Cysts of Giardia intestinalis were found in 9(4.14%) samples, cysts of Entamoeba histolytica in 2(0.9%) samples, eggs of Trituris trichiura in 2(0.9%) samples and oocysts of Cryptosporidium were found in.5(2.30%) samples.&#x0D; Conclusion: Escherichia coli was the most common bacteria and Giardia intestinalis was predominant parasite causing diarrhoea in under 5 children.&#x0D; Bang Med J (Khulna) 2018; 51 : 25-28

Leprosy neuropathy masquerading as cellulitis

Tropical diseases are increasingly reported outside their areas of high prevalence.1 Not only that many physicians are unfamiliar with these conditions but also there is the added challenge that it...

Prevalence of Cryptosporidium species oocysts among small ruminants in Potiskum livestock market, Yobe State, Nigeria

A cross-sectional study was carried out to determine the prevalence of Cryptosporidium species oocysts in small ruminants in Potiskum livestock market, Yobe State, Nigeria. Faecal samples from 203 animals; 100 sheep and 103 goats were tested using modified Ziehl-Neelsen staining technique. An overall prevalence of 20.7% (42/203) was recorded in the study with a higher prevalence of 21.4% (22/103) in goats than in sheep 20% (20/100); although the difference was not significant (p&gt;0.05). In goats, the rate of Cryptosporidium species oocyst was higher in male (27.8%) than in female (22.4%), in young (30.6%) than in adults (22.4%), in those with loose/watery faeces (45.5%) than those with well-formed faeces (22.8%) but the differences were not significant (p&gt;0.05). In sheep, the rate of infection was higher in female (16.9%) than in male (13.0%), in young ((19.2%) than in adults (14.9%) but the differences were not significant (p&gt;0.05). The prevalence was significantly (p = 0.010) higher in sheep with loose/watery faeces (45.5%) than in those with well-formed faeces (12.4%). Breed of sheep did not significantly (p &gt; 0.05) influence the prevalence rates, although the Uda breed had higher prevalence (25%) than the Balami (14.3%) and Yankasa (12.7%) breeds. A similar trend was observed for the sahelian breed of goats (35.7%) compared to the Red sokoto (23.7%), West African dwarf (16.7%) and mixed breeds (10.0%) (p = 0.367). Animals brought to the livestock market from Ngalda town showed highest prevalence of 36.4% than those from the other areas with prevalence rates of (0 to 33.3%). The finding of Cryptosporidium species oocysts in 20.7% of small ruminants in this study underscores the role of this zoonotic parasite diseases in animals and man and loose/watery faeces is a significant factor in the spread among small ruminants.

Occurrence and factors associated with faecal shedding of Cryptosporidium oocysts in small ruminants in Potiskum local government area, Yobe State, Nigeria

Cryptosporidium is an enteric protozoan pathogen associated with neonatal diarrhoea and mortality in small ruminants. Faecal samples of two hundred small ruminants were examined for Cryptosporidium oocysts using formol-ether concentration and modified Ziehl-Neelsen staining technique. Fifty seven (28.5%) out of the 200 faecal samples examined were positive for Cryptosporidium oocysts. The occurrence was higher in sheep (33.0%) than goats (24.0%), in male (32.8%) than female (26.6%), in those with loose/watery faeces (74.4%) than those with well-formed faeces (17.4%), in those between age 4-6 months (39.2%) than 0-3 months (22.2%) and those within 7 months and above (21.1%), in those that drank borehole water (28.7%) than those that drank well/stream water (20.0%), in those raised under semi-intensive/extensive (29.9%) than those under intensive system (21.1%). There was significant association between the occurrence of Cryptosporidium oocysts and age of small ruminants (OR = 2.368, 95% CI on OR: 1.251 &lt; OR &lt; 4.484). On the other hand, there were no significant associations between the occurrence of Cryptosporidium oocysts and sex, breed, faecal consistencies, source of water and management practices of small ruminants (P &gt; 0.05). The species of Cryptosporidium infecting small ruminants identified in this study were C. parvum (50.9%), C. ryanae (26.3%), C. xiaoi (12.3%), C. bovis (8.8%) and C. andersonii (1.8%). The high prevalence of C. parvum observed in this study is of public health importance because of it zoonotic nature. This study shows that age influence the occurrence of Cryptosporidium oocysts and C. parvum is the predominant species in small ruminants in Potiskum, Yobe State.Keywords: Cryptosporidium, Factors, Occurrence, Potiskum, Small ruminants

Investigation of Soil Contamination With Cryptosporidium spp. Oocysts in Different Regions of Yazd, Central Iran

Background: Cryptosporidium species are coccidian parasites that cause gastrointestinal disorders in humans and other animals worldwide. Objective: The aim of this study was to demonstrate the rate of contamination with Cryptosporidium spp. oocysts in soils collected from public parks, primary schools, green areas, kindergartens, suburban areas, streets, residential complexes, backyards and a passenger terminal in Yazd, central Iran. Materials and Methods: This cross-sectional study was conducted from September 2014 to February 2015, and the samples were collected from 9 different study sites and 56 regions. Soil samples were investigated by flotation technique and modified Ziehl–Neelsen staining for Cryptosporidium spp. oocysts. Finally, the slides were examined with a light microscope. The data were analyzed using SPSS software version 20.0 and chi-square statistical test. Results: Of a total of 220 soil samples, 47 (21.36%) were found to contain Cryptosporidium spp. oocysts. Statistical analysis showed that there was no significant difference between the contamination rate and different study sites in Yazd, central Iran (P&gt;0.05). The highest contamination rate was observed in public parks (38.3%) and the lowest in passenger terminal, kindergartens and streets (4.25%) (P=0.934). Conclusion: The results of the present study show that the contamination of soil with Cryptosporidium spp. can be considered a serious problem in Yazd, central Iran. It should be considered particularly in public parks.

First report and molecular characterization of Cryptosporidium spp. in humans and animals in Khartoum state, Sudan.

Background and Aim:Cryptosporidium is recognized to infect several mammalian species as well as humans, causing substantial economic losses and serious public health concern. Infected animals can be a source of environmental contamination and human infections. In general, the occurrence of Cryptosporidium species in animals and human in Sudan and zoonotic importance is not well documented. This study aimed to identify Cryptosporidium spp. infecting different animal species and humans and to compare between different isolates obtained.Materials and Methods:To provide molecular information about Cryptosporidium in animals and humans, both modified Ziehl–Neelsen (MZN) specific stain and molecular assay were used. Concentration techniques followed by three protocols of DNA extraction were carried out. After microscopic screening of 263 fecal samples (goats [n=197], cattle [n=12], sheep [n=12], and human [n=42]), 61 positive and 30 negative, randomly selected samples were used in nested polymerase chain reaction (PCR) targeting part of the 18S RNA.Results:Nested PCR amplification confirmed 91.8% (56/61) of microscopic-positive samples. 8.2% (5/61) of negative samples by PCR (positive by microscopy) were considered false negatives. Sequencing followed by alignment of the 14 isolates indicated that all samples were identical (100%) and belonged to Cryptosporidium parvum.Conclusion:MZN staining procedure is reliable for the routine diagnosis of Cryptosporidium; cetyltrimethylammonium bromide extraction buffer and nested PCR targeting 18S rRNA gene are reliable and useful in epidemiological studies of this parasite.

Cryptosporidium infection in chronic kidney disease patients undergoing hemodialysis in Egypt.

Patients with chronic renal failure are more susceptible to infections due to acquired immunodeficiency caused by uremia. Parasitic infections are one of the significant causes of morbidity and mortality in those patients, So we aimed to assess the prevalence of Cryptosporidium spp. and other protozoan infections in patients undergoing hemodialysis in Qena Governorate, Upper Egypt. The present study took place in Qena University Hospitals, Egypt. Participants were 150 Chronic Kidney Disease (CKD)patients undergoing hemodialysis, and 50 healthy individuals. After questionnaire, three consecutive stool samples from each participant were examined macroscopically and microscopically by different techniques for the presence of different stages of different protozoa. 66% of CKD patients and 26% of the control group were infected with intestinal protozoa. Cryptosporidium spp. were the protozoa with the highest prevalence in cases (40%) and control (6%) with statistical significance (P < 0.05). It was detected only in watery stool samples (P value < 0.05). Residence, age and gender were not significant variables in the prevalence of infection among patients with CKD. In Egypt, few studies had reported the prevalence of Cryptosporidiosis in chronic renal patients. Cryptosporidium infection should be suspected in all cases of prolonged watery diarrhea in CKD patients and stool samples should be examined using special stains as cold modified Ziehl-Neelsen staining for proper diagnosis of Cryptosporidium infections.

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.

Periparturient rise in the Cryptosporidium oocyst count in Beetal goats and evaluation of infection in new born kids

In cryptosporidial infection, very scarce literature is available about periparturient pattern of oocyst excretion in goats and its implications in the epidemiology of disease in kids; thus the present investigation was done. Faecal samples (160) were examined from 20 pregnant goats, collected at weekly interval, commencing from 4 weeks before kidding up to 3 weeks after kidding. Faecal sample examination by Sheather’s floatation sedimentation technique followed by modified Ziehl-Neelsen staining technique revealed an overall positivity of 26.25% for Cryptosporidium oocysts. Prevalence of oocysts excretion was high around parturition and the number of oocysts shed oscillated between 20–400 oocysts/g (OPG) of faeces. Examination of 60 faecal samples of 20 kids born to studied goats from 1 to 3 weeks of age revealed 40% samples positive for Cryptosporidium oocysts and 65% kids became infected by the end of the 3 weeks. The OPG of kids ranged from 100 to 1,560. Molecular characterization by PCR-RFLP of 18S small subunit (SSU) rRNA gene showed that 73.80% and 26.19% goat samples, and 87.5% and 12.5% kids samples were positive for C. parvum and C. ubiquitum, respectively. The study indicated periparturient transmission of Cryptosporidium spp. from pregnant goats to their new born kids.

Improving the microbiological diagnosis of tuberculous meningitis: A prospective, international, multicentre comparison of conventional and modified Ziehl–Neelsen stain, GeneXpert, and culture of cerebrospinal fluid

Tuberculous meningitis (TBM) is the severest form of tuberculosis, but current diagnostic tests are insensitive. Recent reports suggest simple modifications to conventional cerebrospinal fluid (CSF) Ziehl-Neelsen (ZN) staining may greatly improve sensitivity. We sought to define the performance of modified and conventional ZN stain for TBM diagnosis. In hospitals in Vietnam, South Africa and Indonesia we conducted a prospective study of modified ZN with or without cytospin, conventional ZN smear, GeneXpert, and culture on CSF in adults with suspected TBM. A total of 618 individuals were enrolled across 3 sites. Compared with the TBM clinical diagnostic gold standard for research (definite probable or possible TBM), sensitivity of conventional ZN and modified ZN with cytospin were 33.9% and 34.5% respectively (p = 1.0 for the difference between tests), compared with culture 31.8% and Xpert 25.1%. Using culture as a reference, sensitivities of conventional ZN, modified ZN with cytospin, and Xpert were 66.4%, 67.5%, and 72.3%, respectively. Higher CSF volume and lactate, and lower CSF:blood glucose ratio were independently associated with microbiologically confirmed TBM. Modified ZN stain does not improve diagnosis of TBM. Currently available tests are insensitive, but testing large CSF volumes improves performance. New diagnostic tests for TBM are urgently required.

Effects of Echinacea Purpurea on Cryptosporidiosis in Immunosuppressed Experimentally Infected Mice

AbstractBackground: Cryptosporidium species are worldwide coccidian parasites. They are considered the second cause of diarrhea and death in children after rotavirus. Current treatment options for cryptosporidiosis are limited. There is an urgent need to develop new anti-cryptosporidial agents.Aim of the Study: To assess the activity of Echinacea purpurea in treatment of experimental cryptosporidiosis in immunosuppressed mice.Methods: Ninety mice were immunosuppressed using oral dexamethasone and divided into 5 groups. Echinacea was used as 100 mg/kg/day on day 15 post infection for five consecutive days. Stool samples from all survived mice were subjected to modified Ziehl-Neelsen staining. All mice were sacrificed for histopathological examination and immuno-histochemical staining of their ilea sections for IL-17 and Cox-2.Results: The least mortality rate (0%), the least oocysts shedding (1.10±2.31), the least endogenous developmental stages (3.50±2.24), the most improved pathological changes and the highest cure rate (90%) were observed in mice treated with Echinacea/nitazoxanide combination. Moreover, combi-nation therapy significantly reduced IL-17 and Cox-2 expres-sion in ileum sections compared to the positive control group. Echinacea monotherapy significantly reduced fecal oocyst shedding and ileal endogenous developmental stages with improved pathological changes compared to the positive control group. Echinacea increased the cure rate with no significant difference when compared to nitazoxanide. It significantly decreased IL-17 and Cox-2 in ileum sections compared to the positive control group.Conclusion: Echinacea purpurea/nitazoxanide combina-tion represents significant advances in treatment of experi-mental cryptosporidiosis infection in immunosuppressed mice.