Modified Ziehl-Neelsen Staining Method Research Articles

Occurrence of Cryptosporidium Oocysts in Faeces, Milk and Water Sources in Sedentary Fulani Herds in Selected Local Government Areas of Kaduna State, Nigeria

Abstract Cryptosporidiosis is a neglected tropical zoonotic disease, commonly associated with a diarrhoea, caused by a protozoan parasite of the genus Cryptosporidium. The aim of this study was to determine the occurrence of Cryptosporidium oocysts in faeces, milk, and water sources, and its associated risk factors in sedentary Fulani herds. Seven hundred and fifty faecal samples (240, 180, 240 and 90 from cattle, sheep, goats and dogs, respectively), and 120 samples each, of milk and water were collected for this study. Faecal samples were concentrated by formol ether concentration technique, then stained using Modified Ziehl Neelsen staining method, to detect Cryptosporidium oocysts. Cryptosporidium oocysts in milk and water samples were also concentrated and stained with same stain. The prevalence of Cryptosporidium oocysts in (cattle, dog, sheep and goat faeces were 18.3 %, 15.6 %, 13.9 % and 4.2 %, respectively.), Significantly higher prevalence was observed in calves ≤ 1 year (P = 0.002), female cattle (P = 0.007) and animals with poor body condition (P < 0.001) than in the other groups. Goats with poor body condition scores had significantly higher prevalence (P = 0.008) than other groups while dogs ≤ 6 months of age also had significantly higher prevalence (P = 0.03) than older dogs. The prevalence of Cryptosporidium oocysts in bulk cow milk was 11.7 % while a prevalence of 13.3 % was observed in the water bodies sampled. Hence the findings of this study are of great public health significance, therefore, inhabitants of these LGAs should be informed and educated on the need for improvement of sanitary measures during the course of milking these animals, and the need for adequate pasteurization, of milk before consumption.

Prevalence and associated risk factors of Cryptosporidium infection in calves and hospitalized humans in Libo Kemkem, North Western Ethiopia.

Cryptosporidium infection is one of the major causes of acute gastroenteritis and diarrhoea caused by a protozoan parasite affecting vertebrates and humans. The disease is prevalent in cases of immunocompromised individuals. Despite the impact of the diseases in calf and hospitalized humans, well-documented studies are not available in the study area. The objectives of this study were to determine the prevalence of Cryptosporidium infection in calves and hospitalized humans and assess the major associated risk factors associated with Cryptosporidium infection in calves and hospitalized humans. A cross-sectional study was conducted from November 2020 to March 2021 on calf and human Cryptosporidium infection in Libo Kemkem District, North West Ethiopia. A total of 193 calves and 122 human stool samples admitted to the hospital were used for this study. Three kebeles were selected purposely, and individual calves were selected using a simple random sampling method. A number of sampled calves were allocated proportionally to the selected kebeles. Human samples were collected using a systematic random sampling method. Faecal and stool samples were examined using a modified Ziehl-Neelsen staining method. The overall prevalence of calf and human Cryptosporidium infection found in this study was 15.5% and 11.5%, respectively. Age of calf, breed, body condition, water source, faecal consistency and hygienic condition were found significantly (p<0.05) associated with Cryptosporidium infection in the calf. Similarly, the source of potable water, immunocompromisation and contact with domestic animals were found to be significantly (p<0.05) associated with Cryptosporidium infection in humans. There was a higher prevalence of Cryptosporidium infection in calves and humans in Libo Kemkem District. Therefore, the implementation of proper prevention methods of zoonotic Cryptosporidium infection between calf and human beings through significant risk factors is mandatory. Furthermore, additional studies to investigate the levels of economic importance of the disease should be conducted.

Prevalence of Cryptosporidium Infection among Children Attending Maryam Abacha Women and Children Hospital, Sokoto State, Nigeria

Study's Excerpt/Novelty This study investigates the prevalence of Cryptosporidium among children attending Maryam Abacha Women and Children Hospital, Sokoto, using Formol ether concentration and Modified Ziehl Neelsen staining methods. It identifies a high prevalence of (71.1%) and shows a strong correlation between Cryptosporidium infection, diarrhea, and source of drinking water. These findings underscore the necessity for routine diagnostic testing of Cryptosporidium among children for improved detection, treatment, and overall patient outcomes in the region. Full Abstract Cryptosporidium infection is ranked as the second most common cause of diarrheal disease and fatalities among children in developing nations. Nonetheless, due to the fact that many healthcare providers do not consistently request Cryptosporidium diagnostic tests, the prevalence of cryptosporidiosis is probably underestimated. The current study is therefore aimed at investigating the prevalence of cryptosporidiosis among children attending Maryam Abacha Women and Children Hospital (MAWCH), Sokoto. During the timeframe of April to June 2021, stool samples were collected from 114 diarrheic and non-diarrheic children and analyzed using Formol ether concentration and Modified Ziehl Neelsen staining methods. Out of 114 samples analyzed, 81(71.1%) were positive for Cryptosporidium oocysts. A higher prevalence of Cryptosporidiosis was observed in males 45(39.5%) compared to females 36(31.6%) participants and higher 41(36.0%) in younger children aged 0-1 years than the older children. Similarly, a greater prevalence of Cryptosporidiosis was observed in diarrheic children (42.1%) than in non-diarrheic children (28.9%). A statistically significant relationship was observed between the occurrence of Cryptosporidium and diarrhea (P = 0.002) and the source of drinking water (P = 0.001). Routine diagnosis of Cryptosporidiosis among children is vital and can enhance effective diagnosis, treatment, and consequently, improvement in the patients' well-being.

Identification of major protozoal enteropathogens causing calf diarrhea in dairy farms in and around Holeta Town, Oromia Special Zone, Ethiopia

Calf diarrhea can be caused by a variety of pathogens, including bacteria, viruses, protozoa, and intestinal parasites. Giardia, Eimeria, and Cryptosporidium are the most significant protozoan parasites and are all individually and ollectively infectious. A case-series study was carried out in and around Holeta dairy farms, Oromia Special Zone, Ethiopia, from November 2017 to April 2018 to identify the main protozoan enteropathogens from diarrheic calves. Samples were purposely collected from three dairy farms: Serkalem Dairy Farm (SDF), Holeta Agricultural Research Centre (HARC), and Ada Berga Agricultural Research Centre (ABARC). A total of 93 fecal samples were taken from diarrheic calves of up to 4 months of age. Protozoan enteropathogens were identified using flotation and modified Ziehl Neelsen staining methods. Cryptosporidium, Eimeria, and Giardia were detected in 44 (47.3%), 50 (53.8%), and 34 (36.6%) of the diarrheic fecal samples examined, respectively. The findings indicated that there were 17 (18.27%) cases of Cryptosporidium, Eimeria, and Giardia as mixed infections, 16 (17.2%) cases of Cryptosporidium and Eimeria, 5 (5.4%) cases of Cryptosporidium and Giardia, and 4 (4.3%) cases of Eimeria and Giardia, compared to 13 (13.97%), 6 (6.45%), and 5 (5.4%) cases of Cryptosporidium, Eimeria and Giardia as single infections, respectively. The selected farms had significant prevalence levels of three common protozoa: Eimeria, Giardia, and Cryptosporidium. Further molecular research is required to identify the species and genotype levels of protozoal enteropathogens and related risk factors.

Assessment of Co-Infection in Tuberculosis Patients Prior to their Enrollment to the Direct Observed Treatment (Dot) Program in Buea, Limbe, and Douala Tuberculosis Treatment Centers

Introduction: Co-infections during Tuberculosis infection are among the leading causes of morbidity and mortality worldwide. These infections are a major public health problem among patients in most developing countries, including Cameroon. One of the important public problems in coendemic areas is co-infection with TB in humans. Moreover, there is no adequate research hitherto that provides information on the level of co-infection in Tuberculosis patients. Therefore, this study aimed to assess the prevalence and intensity of co-infections in newly diagnosed Tuberculosis patients prior to their enrollment to the Direct Observed Treatment (DOT) program in Buea, Limbe and Douala Tuberculosis treatment centers. Methods: This cross-sectional study was conducted from March to November 2020 using a purposive sampling technique. A fresh sputum sample was collected, processed, and examined through a smear microscopy and culturing technique. Blood samples were collected through finger pricking for the detection of blood parasites using thin and thick blood smears. Stool samples were also collected and processed using a direct wet mount, and Kato-Katz and Modified Ziehl Neelsen staining methods. The data was analyzed using SPSS version 20.0. A Chi-square test was done to assess the prevalence of co-infection among all TB patients recruited for this study. Results: A total of 200 newly diagnosed TB-positive patients were recruited from the three study sites. Out of the total participants recruited, 29.0% were diagnosed positive for blood parasites, 2.0% for intestinal parasites and 15.0% for viral infections, bringing the overall prevalence of co-infection to 46.0%. When specific infections were considered, malaria, filaria, intestinal parasites, HIV and HBV constituted 28.0%, 1.0%, 2.0%, 14.5% and 0.5% respectively. When dual-infection was considered, the results were 0.5% malaria/filaria, 1.5% malaria/intestinal parasites and 2.0% malaria/HIV. The mean parasite densities for all samples diagnosed positive for malaria, filarial and intestinal parasites were 107.0 ± 338.8 parasites/uL, 25.7 ± 356.4 mf/mL and 85.0 ± 85.4 EPG, respectively. Conclusion: The results indicate that there is an overlap of co-infections in TB patients with their interactions expected to lead to complications and difficulty when managing the TB patients, calling for more research and recommending more assessment for co-infection in TB patients before starting treatment.

Cryptosporidium in human-animal-environment interphase at Adama and Asella areas of Oromia regional state, Ethiopia

BackgroundIn Ethiopia, several studies have reported the occurrence of Cryptosporidium infection in young calves, lambs, and HIV positive patients. However, research on cattle and sheep of all ages, as well as humans, has been limited.MethodsA cross-sectional study was conducted to investigate the occurrence of Cryptosporidium in dairy cattle, sheep, humans, and manure samples. Dairy cattle and sheep were stratified according to their age groups and randomly sampled. Stool samples from volunteered dairy farm workers and rural household members were collected purposefully. A semi-structured questionnaire was also used to collect information about manure handling practices and socio-demographic variables. All collected samples were processed with the sugar floatation technique and the modified Ziehl–Neelsen staining method. Logistic regression was used to analyze the association of risk factors with the occurrence of Cryptosporidium in study subjects (p < 0.05).ResultsThe overall prevalence of Cryptosporidium in dairy cattle, sheep, humans, and manure samples was 20.5%, 14%, 16%, and 13.20%, respectively. Cryptosporidium infection was significantly higher in dairy cattle aged 1–12 months and 13–36 months with odds of 3.48 and 3.07 times higher, respectively, compared to others. Similarly, its occurrence was 2.69 times higher in sheep aged 1–6 months than those above 6 months. And also, a higher average oocyst count (above 10 oocysts) per-field was observed in cattle aged 1–12 months, followed by sheep aged 1–6 months. Furthermore, the likelihood of infection was 13 times greater in farm workers compared to household members of smallholder farmers. In addition, the occurrence of oocysts was 22.8 times higher in manures from dairy cattle than that of sheep. About 16.8% of the study respondents had manure disposal pit, 98.1% of them used manure as fertilizer for crop and vegetable production without any treatment.ConclusionsThe study revealed the occurrence of Cryptosporidium infection in all age groups of dairy cattle and sheep, humans engaged in animal production. Occurrence of Cryptosporidium in manure suggests it potential contamination of environment and water sources.

Prevalence of Cryptosporidiosis and It’s Associated Risk Factors Among Human Population in Zakho District, Duhok Province, Kurdistan Region, Iraq

Cryptosporidiosis caused by species belonging to genus Cryptosporidium. Cryptosporidiosis is a zoonotic disease that affects both animals and humans, making it a major public health and veterinary concern. A cross sectional study was performed during the period from August 2021 to March2022, to estimate the prevalence of Cryptosporidium spp. in the human population in order to determine the effect of associated risk factors on the rate of infection in Zakho district, Duhok. Seven hundred Fecal specimens were collected from Zakho General Hospital and two primary schools located in suburban area of Zakho city. The collected stool specimens were examined macroscopically and microscopically using a modified Ziehl–Neelsen staining method. Cryptosporidium infection was found in 87.57% (613/700) of the examined specimens. Males showed slightly higher infection rate than females (88.4% vs. 85.5%) with non-significant difference (P &gt; 0.05). Ages group (31-41 years) showed highest rates, with significant difference (P&lt;0.05). The highest rates of infection were reported among families with 6-10 members, people drinking unfiltered municipal tap water, suburban residents, those with close contacts with domestic animal, and illiterate people. The control of this disease necessitates the improvement of living conditions, providing potable clean water and disseminating health education programs among the community.

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.

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.

Clinical and Laboratory Diagnosis of Cryptosporidiosis among Children with Acute Gastroenteritis at a Tertiary Hospital, Cairo, Egypt

Most studies on gastroenteritis have focused on viral and bacterial infections, while gastroenteritis where intestinal protozoan parasites may have played a role has not been well studied. This study was therefore, designed to assess the frequency and several potential risk factors for Cryptosporidium infection among children suffering from acute gastroenteritis and presented to a tertiary hospital in Cairo, Egypt. Effectiveness of modified Ziehl-Neelsen (MZN) and nested polymerase chain reaction (nPCR) for Cryptosporidium detection were evaluated as well. A cross-sectional study was performed during the period from July 2018 to December 2018, where 100 human diarrheic stool samples were collected from children aged 3 months up to 12 years old presented to Ain Shams University Pediatrics Hospital, Cairo, Egypt with acute gastroenteritis. Demographic and clinical data were obtained from the participants. Initial parasite screening was done using the MZN staining method, and microscopically examined for Cryptosporidium infection, while genotyping was based on molecular diagnostic assays using nPCR and sequencing for selected samples. The overall frequency of Cryptosporidium infection was 5% using light microscopy, while 19% of samples were positive by nPCR. Cryptosporidium hominis was the only detected genotype. Clinical picture among cases were not significant in comparison to patients with other causes of gastroenteritis. Cryptosporidium infection is more common below 5 years of age; however, clinical data are not enough for suspicion of infection. Nucleic acid-based methods are more sensitive and specific despite the high cost in developing countries. However, real estimation of Cryptosporidium disease burden is of an outmost importance to achieve prevention and detection of the Cryptosporidium species genetic diversity. Lay summaryCryptosporidium is a protozoan, which causes gastroenteritis in humans. It is most common below 5 years of age; however, diarrhea and vomiting characteristics are not different from other causes of gastroenteritis. General diagnostic methods are inadequate for detection of these infections. Nested polymerase chain reaction (nPCR) and sequencing are accurate methods for pathogen detection and species verification. Our study included 100 Egyptian children with acute gastroenteritis. The overall frequency of Cryptosporidium infection was 5% using light microscopy, while 19% of samples were positive by nPCR. The clinical picture of the children presenting with this disease was not significantly different from those presenting with gastroenteritis due to other causes. This emphasizes the importance of proper diagnosis to know the true burden of the disease.

Molecular Detection of Brucella melitensis Associated Reproductive Failure in Native Sheep Flock: Potential Public Health Pathogen from Tropical Region of Tamil Nadu

Brucellosis remains a global burden since the last century. In India, both animal and human brucella infections drastically increase every year. Outbreaks of 40 ewe’s abortion were investigated in native sheep’s from the tropical region of Tamil Nadu. Aborted maternal and fetal tissues were initially subjected to Stamps Modified Ziehl Neelsen (S-ZN) staining method. The DNA was extracted and screened for Brucella genome by Brucella cell surface salt extractable proteins (bcsp31) gene based PCR assay. Further, PCR positive samples were subjected for AMOS PCR for B. abortus, B. ovis, B. melitensis and B. suis identification and differentiation. S-ZN staining of aborted tissues revealed pink, coccobacilli organisms giving presumptive identification of Brucella sp. Molecular detection of brucellosis by bcsp31 gene based PCR in placental tissues and fetal abomasal contents of both the animals yielded a specific amplicon of 223 bp confirming it as Brucella genus infection. Molecular detection and differentiation by AMOS PCR revealed a specific amplicon of 731 bp indicating the involvement of B. meletensis infection in the native free-ranging sheep flock. It is a potent public health pathogen, highly essential to be monitored for the strategic control of brucellosis.

Assessment of Cryptosporidium in Patients With Gastroenteritis by Modified Ziehl-Neelsen Staining Method in East Azerbaijan Province of Iran During 2018-2019

Introduction: Cryptosporidiosis is a worldwide zoonotic disease that is caused by Cryptosporidium species and leads to acute or chronic diarrhea and vomiting in patients. Due to the suitable conditions of East Azerbaijan province for parasite transmission, the present study was performed to evaluate the prevalence of Cryptosporidium infection in patients with gastroenteritis referred to hospitals and reference laboratories of East Azerbaijan province during 2018-2019. Methods: A descriptive cross-sectional study was performed on patients with diarrhea who referred to hospitals and reference laboratories of East Azerbaijan province during 2018-2019. In this study, 180 fecal samples were collected from patients with gastroenteritis by cluster-random sampling. Stool samples were stored in 10% formalin and transferred to a parasitology laboratory and examined for parasite contamination by modified acid-fast staining (Modified Ziehl-Neelsen) method. Data were analyzed by chi-square test using SPSS. Results: Out of 180 samples, the highest number of diarrhea samples belonged to the age group over 40 years (35.5%) and the lowest belonged to the age group of 10-20 years (7.7%). Additionally, 56% of subjects were male and 44% were female. Of these patients, 57.7% lived in urban areas and 42.3% in rural areas. Cryptosporidium was observed in 1.6% (3 cases) of patients with gastroenteritis. There was a statistically significant relationship between Cryptosporidium infection and the age of patients with diarrhea (P&lt;0.05). Conclusion: In the present study, a lower prevalence of cryptosporidiosis has been determined in comparison with previous studies in East Azerbaijan province. However, Cryptosporidium and other parasitic infections in the feces of patients with diarrhea referred to hospitals or reference laboratories need to be carefully diagnosed by appropriate parasitological methods.

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

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

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

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

Histopathological changes of brucellosis in experimentally infected guinea pig

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

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.

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..

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.

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.

Prevalence and risk factors of intestinal protozoan infections: a population-based study in rural areas of Boyer-Ahmad district, Southwestern Iran.

BackgroundParasitic infections are still a significant health problem in rural areas in developing countries including Iran. There is no recent population-based data about the prevalence of human intestinal parasites in most rural areas of Iran. The current study aimed to determine the prevalence of intestinal protozoan infection in inhabitants of rural areas of Boyer-Ahmad district, Southwestern Iran.MethodsA total of 1025 stool samples were collected from the inhabitant of 50 randomly selected villages in Boyer-Ahmad Township. The stool samples were evaluated by parasitological methods including, direct wet-mounting, formalin ethyl acetate concentration, zinc sulfate floatation, and Trichrome permanent stain for detection of protozoan infections. Diarrheic samples were further evaluated with a modified Ziehl–Neelsen staining method for detection of coccidian parasites.ResultsThe prevalence of both pathogenic and nonpathogenic intestinal parasites in the population was 37.5% (385 out of 1025 cases), some individual with multiple infections. Giardia lamblia was detected in 179 (17.46%), Blastocystis hominis in 182 (17.76%), Entamoeba histolytica/dispar in 9 (0.87%), Endolimax nana in 216 (21.07%), Entamoeba coli in 151 (14.73%), Ioedamoeba butschlii in 45 (4.39%), Chillomastix mesnili in 22 (2.14%), Trichomonas hominis in 2 (0.19%) and Dientamoeba fragillis in 2 (0.19%) of cases. Multivariate logistic regression revealed significant associations between protozoan infection (pathogenic protozoa) and contact with animals (OR yes/no = 2.22, p < 0.001) and educational status (OR higher/illiterate = 0.40, P = 0.01).ConclusionFindings of this study demonstrated that protozoan infection rate in rural areas of southwestern Iran is still high and remained as a challenging health problem in these areas.