Ziehl-Neelsen Technique Research Articles

Presence of Intestinal Parasites in Patients with Chronic Non-Communicable Diseases in Masaya (Nicaragua).

A cross-sectional study was conducted in Masaya (Nicaragua) to estimate the prevalence of intestinal parasite (IP) infections in patients with non-communicable diseases (NCDs) and to determine the associations between the types of NCDs and patients' epidemiological characteristics of infection. A total of 157 preserved faecal samples were examined (direct wet mount, formalin/ethyl acetate concentration and modified Ziehl-Neelsen technique). Microscopically positive faecal sample identification was completed by conducting a molecular study. The total prevalence of IP was 52% in NCD patients. Diabetic patients presented an IP prevalence of 42%. Blastocystis presented the highest prevalence (42%). A molecular analysis of Giardia intestinalis (prevalence of 1.3%) revealed 100% of sub-assemblage BIII and the Entamoeba complex (5%) was identified as E. dispar. Blastocystis ST1 appeared in 44% of those suffering from diabetes and ST3 in 66% of those suffering from hypertension, while ST2 only appeared in those suffering with several NCDs simultaneously. In diabetic patients, the risk of infection is associated with having pets (p = 0.021) and land-floor houses. The risk of infection appears to be statistically related (p = 0.019) in those with several NCDs having received a previous helminthic deworming treatment. Coordinated public health activities for IP and NCD screening and diagnosis are crucial to their successful control programmes.

Prevalence of gastrointestinal parasites in domestic pigs from family farms in northeastern Argentina

Domestic pigs serve as significant hosts and reservoirs for multiple parasite species, some specific to pigs and many others of zoonotic importance. This study aimed to assess the prevalence of intestinal parasites in pigs within a rural area in northeastern Argentina. We also examined demographic information, breeding conditions, and exposure factors associated with parasite presence. Pig feces were subjected to coprological examination through flotation and sedimentation techniques. Modified Ziehl-Neelsen technique was employed to examine oocysts of Cryptosporidium spp. In total, 29 family farms with pig pens were analyzed, and 42 stool samples were collected from pigs on these farms. At the farm level, the presence of at least one parasite species was recorded in 27 rural houses (93.1%). We found that 90.4% of pigs were parasitized, with a specific parasitoses of 10 species, with a maximum of six species in a single host. The most prevalent protozoa were Entamoeba spp. (57.1%) and Blastocystis sp. (45.2%), followed by Iodamoeba butschlii (33.3%), Neobalantidium coli (21.4%), coccidia (14.2%), Cryptosporidium spp. (9.5%) and Giardia spp. (2.3%). The most prevalent helminths were Strongylidae eggs (52.3%), Ascaris spp. (14.2%) and Trichuris spp. (2.3%). We advocate for an urgent need to implement a comprehensive prophylaxis program prioritizing general hygiene practices such as regular cleaning, removal of fecal material and renewal of drinking water. Additionally, vaccination and deworming protocols should be implemented. Furthermore, this study highlights the necessity for molecular-level evaluations to detect potential zoonotic genotypes of the identified protozoa.

Role of Fluorescent Staining and Ziehl-Neelsen Techniques in The Detection of Mycobacterium Tubercle Bacilli in Lymph Node Aspirate

Background Tuberculosis is the important disease of infectious etiology especially in the developing nations. It is a major health issue in India. FNAC is an economic and rapid tool having high diagnostic accuracy for LN tuberculosis. Present study was conducted to describe the role of fluorescent staining and Ziehl- Neelsen techniques in detecting Mycobacterium tubercle bacilli on FNAC samples of tubercular lymphadenitis.MethodsThis prospective study was carried out at Department of Pathology, Muzaffarnagar Medical College, U.P. It included 100 patients having enlarged lymph nodes which were suspected of TB. After FNAC, cytological examination was done and smears prepared were stained with ZN stain and Auramine Rhodamine (AR) stain.ResultsTwenty eight (28.0%) cases were found in 21-30 years age group followed by 20 cases (20%) in 31- 40 year age group and 11-20 year age group. Most were females forming female to male ratio 1.7:1. Cytomorphologically, four patterns were observed– Necrotizing granulomatous lymphadenitis in 41% cases, Granulomatous lymphadenitis in 29% cases, only Necrotizing lymphadenitis in 19% cases and Necrotizing lymphadenitis with suppuration in 11% cases. ZN staining showed AFB in 35% cases while AFB was not seen in 65% cases. Out of these 65% cases, fluorescent staining was positive in 29%, indicating significant association of fluorescent staining with ZN method. Conclusions Only with the cytological findings of lymph node, it is difficult to arrive at confirmatory diagnosis of TB. Fluorescence Microscopy (FM) increases the sensitivity for diagnosing TB in underdeveloped nations with high tuberculosis prevalence, such as India.

Zoonotic parasite contamination of some green, leafy and spicy vegetables sold in major markets of Makurdi metropolis, Benue state

Consumption of contaminated vegetables can result in parasite ingestion with subsequent health problems. The spread of zoonotic parasites through contaminated vegetables is a complicated problem that requires attention to ensure food safety and protect public health. This study investigated the zoonotic parasitic contamination of some green, leafy, and spicy vegetables sold in Makurdi metropolis, Benue State. A cross-sectional study was designed in which four hundred green, leafy, and spicy vegetables consisting of five different vegetables, were collected from five markets in Makurdi town, Benue state. Sediments obtained by washing the vegetables with normal saline were centrifuged at 1500 rpm for five minutes before being subjected to flotation, sedimentation, and modified Ziehl-Neelsen techniques, and viewed under the microscope at various magnifications. The overall prevalence of zoonotic parasites was 59.75%. Strongyloides spp. larvae (40.5%), Cryptosporidium spp. Oocyst (0.5%), Balantidium coli cyst and trophozoite (7%), Ascaris spp (5.75%), Taenia spp. (9%), Strongyle ova (4.5%), and Giardia lamblia cyst (1.5%) were identified. Results showed a significant relationship between vegetable type and parasite contamination of vegetables (p < 0.05), but no significant relationship between market location and parasite contamination (p > 0.05). The high prevalence of zoonotic parasites observed in this study highlights the potential health risks associated with consuming contaminated vegetables. The study recommends public enlightenment of vegetable sellers and consumers in Makurdi on the importance of hand hygiene and safe handling of vegetables to prevent exposure to pathogenic microorganisms.

Detection of <I>Giardia</I> cyst and <I>Taenia</I> eggs in river water samples collected from Aleto River in Eleme Local Government Area, Rivers State, Nigeria

This study aimed at detecting Giardia cyst and Taenia eggs in river water samples collected from Aleto River in Eleme Local Government Area, River State, Nigeria, to check for potability and purity of water.Water samples of river water were collected from two points (downstream and midstream) of Aleto river. Labelled clean sample bottles with tightly-fitted lids were used to collect water samples from the river and stored in coolers with ice packs to maintain their temperature and transportation to the laboratory. Collected water samples were transferred into sterile vial tubes and centrifuged at 2000 rpm for five minutes, after which the supernatant fluids were discarded, while the sediments smeared using spatula on labelled slides stained with specific dyes, using the wet mounts, trichome stain technique, and modified Ziehl–Neelsen techniques, and viewed under the light microscope to identify the presence of parasites.This study showed the presence of ten trophozoites of Giardia lamblia and ten eggs of Taenia species isolated from the river water samples collected. These findings highlight the potential health risks associated with waterborne parasites and emphasizes the importance of effective water quality management to safeguard public health. Furthermore, the observation of parasites underscores the need for comprehensive water quality assessment and monitoring. This study highlights the contamination of the river with parasitic organisms and urgently needs effective water management strategies to mitigate the risks to public health. Vital interventions encompass enhancing sanitation practices, curbing fecal contamination, and augmenting water quality surveillance mechanisms.

Intestinal parasitic infections and predisposing factors among prison inmates in Southern Ghana: A cross-sectional study.

The environment within prisons, such as overcrowding, lack of access to portable water, poor sanitation, and hygiene predisposes inmates to infections, including intestinal parasitic infections (IPIs). This study therefore determined the prevalence and associated factors of IPIs among prison inmates in Southern Ghana. A cross-sectional study using the stratified sampling technique was employed. The study recruited 461 prison inmates across three notable prisons, with 50 inmates from Ho Central, 357 inmates from Nsawam Medium Security, and 54 inmates from Sekondi Central Prisons all in Southern Ghana. A structured closed-ended questionnaire was administered to collect data on sociodemographics, lifestyle/behavioral characteristics, and signs and symptoms of IPIs. In addition, stool samples were collected and analyzed for the presence of various stages of intestinal parasites (trophozoites, cysts, ova, and larva) using formol-ether concentration, and the modified Ziehl-Neelsen techniques. The prevalence of IPIs among inmates of the three selected prisons was 38.2% (95% confidence interval [CI]: 33.72%-42.79%). The prevalence of IPIs among inmates of the three prisons, the Ho Central, Nsawam Medium Security, and Sekondi Central were 46% (95% CI: 31.82%-60.68%), 37.5% (95% CI: 32.49%-42.79%), and 35.2% (95% CI: 22.68%-49.38%), respectively. After adjusting for confounders, the following factors; handwashing after defecation (adjusted odds ratio [AOR]: 0.05 [0.00-0.67]; p = 0.024), trimmed fingernails (AOR: 0.32 [0.13-0.76]; p = 0.011), itchy skin (AOR [95% CI]: 5.99 [3.43-10.43]; p < 0.001), anal itching (AOR [95% CI]: 0.35 [0.19-0.62]; p < 0.001), nausea (AOR [95% CI]: 5.57 [3.22-9.65]; p < 0.001), and worm expulsion (AOR [95% CI]: 3.80 [1.42-10.18]; p = 0.008) were found to be associated with intestinal parasitic infections. The study revealed that the prevalence of intestinal parasitic infections among prisons in Southern Ghana is high and is therefore an important public health concern. The Public Health Department of Ghana Health Service should therefore embark on health promotion and deworming exercise in all prisons in Ghana.

Utility of Conventional ZN Staining and Modified Bleach Techniques in Detection of Acid Fast Bacilli from Aspirates of Suspected Cases of Tuberculous Lymphadenitis

Introduction: Tuberculosis (TB) is a significant public health problem in many developing countries, including India. Diagnosing tuberculous lymphadenitis, in particular, poses challenges as it requires the demonstration of Acid-Fast Bacilli (AFB) using the conventional Ziehl-Neelsen (ZN) technique. The Modified Bleach Technique, which involves liquefying the aspirated specimen with sodium hypochlorite followed by centrifugation, has been described to enhance the yield of AFB, improving sensitivity and diagnostic accuracy. Aim: To highlight the role of the bleach concentration method compared to conventional ZN staining in detecting AFB in Fine-Needle Aspiration (FNA) material from clinically suspected cases of tuberculous lymphadenitis. Materials and Methods: A prospective cross-sectional study was conducted at the Department of Pathology, MVJ Medical College and Research Hospital, Bengaluru, Karnataka, India from November 2020 to December 2022. Fine needle aspiration material from 100 cases of clinically suspected tuberculous lymphadenitis was included. In each case, part of the aspirate was processed for ZN staining, while the remaining material was used for modified bleach staining and Cartridge Based Nucleic Acid Amplification Test (CBNAAT). Patient data, including age, sex, duration and site of swelling, nature of aspirate, and cytomorphological diagnosis, were documented. The data was entered into a Microsoft excel spreadsheet and analysed using the Chi-square test, with a p-value&lt;0.05 considered statistically significant. Results: The age range of the study participants was 1-80 years. Out of 100 cases, 13 were excluded due to inadequate material. The most common lymph node site was cervical 69 (79.3%), followed by axillary 8 (9.1%) and supraclavicular 6 (6.89%). Among the 87 cases, reactive lymphadenitis was diagnosed in 36 (41.3%), suppurative lymphadenitis in 12 (13.8%), necrotising granulomatous lymphadenitis in 12 (13.8%), granulomatous lymphadenitis in 15 (17.3%), and necrotising lymphadenitis in 12 (13.8%). Out of the 100 cases included in the study, 32 cases were confirmed as tubercular lymphadenitis by CBNAAT. Among these 32 cases, conventional ZN stain was positive for AFB in only 3 cases (9%), whereas the Modified bleach stain was positive in 16 cases (50%). The results were compared with CBNAAT findings, and the modified bleach technique showed sensitivity (59.4%), specificity (100%), Positive Predictive Value (PPV) (100%), Negative Predictive Value (NPV) (80%), and Accuracy (85%). Conclusion: The Modified bleach method of ZN staining is a simple, safe, inexpensive, and easy-to-perform technique. With the bleach method, AFB was easily visible and detectable in the majority of AFB-positive cases. Additionally, the bleach method preserved the morphology of AFB better. It improves the microscopic detection of AFB, leading to a definitive diagnosis of TB on cytology.

Red cell parameters, iron, vitamin B12 and folate levels of pulmonary tuberculosis patients attending clinic at General Hospital, Akamkpa, Cross River State, Nigeria

Introduction: Tuberculosis caused by Mycobacterium tuberculosis, is a disease of public health importance, characterized by a chronic granulomatous inflammation in humans. In this study, red cell and iron parameters as well as vitamin B12 and folate levels were assessed with a view to investigate the presence or absence of iron deficiency and/or vitamin B12 and folate deficiency in tuberculosis disease Methods: Fifty (50) male and female tuberculosis patients within the ages of 15-60 years and attending clinic at the General Hospital, Akamkpa were enrolled in this study. Fifty apparently healthy and mantoux-negative subjects who were age and gender matched and resident in Akamkpa served as control. Ethical approval and informed consent were obtained from the Cross-River State Ministry of Health and all participants. Demographic information was obtained by face-to-face interview. Diagnosis of TB was by the Ziehl-Neelsen technique. Red cell and iron parameters, vitamin B12 and folate levels were determined by standard methods. Data were analyzed using student t-test on statistical package for social sciences version 21and a P value less than or equal to 0.05 is considered significant. Results: The mean age of TB patients (34.7±10.4 years) is comparable to that of the control (32.4±8.6 years) with more males (56%) affected than females (44%). Fifteen (30%) of the TB patients have primary level of education with 31 (62%) and 4 (8%) having attained secondary and tertiary levels. The level of education for the control was 12 (24%), 22 (44%) and 16 (32%) for primary, secondary and tertiary respectively. The TB patients were farmers 20 (40%), traders 15 (30%), students 9 (18%) and civil servants 6 (12%) while the controls consisted of 14 (28%) farmers, 16 (32%) traders, 12 (24%) students and 8 (16%) civil servants. The packed cell volume and haemoglobin concentration of TB patients (0.34±0.05 L/L and 121.4±11.3 g/L) was significantly lower (p &lt; 0.05) than 0.41±0.04 L/L and 145.2±13.1 g/L for the control. The red cell count and mean cell volume of TB patients (4.55±0.73 x 10 12/L and 79.21±2.40 fl) was comparable (p &gt; 0.05) to control values (4.84±0.54 x 10 12/L and 80.18±1.30 fl). Mean cell haemoglobin and mean cell haemoglobin concentration of TB patients was 27.58±2.12 pg and 34.37±1.43 g/dl, which was significantly lower than 29.72±2.24 pg and 35.73±1.38 g/dl obtained for control. The serum iron of TB patients (47.90±6.25µg/dl) was significantly lower (p = 0.001) than control value 109.03±8.56µg/dl. The total iron binding capacity (188.05±33.01µg/dl) and transferrin saturation with iron (28.00±4.54 %) were significantly lower (p = 0.001) for TB patients versus control (252.28±36.30µg/dl and 42.46±6.23%). Serum ferritin of TB patients (345.30±82.61 ng/ml) was significantly higher (p = 0.001) when compared to the value for control (108.62±28.50 ng/ml). Vitamin B12 and folate levels (245.37±39.62 ng/L and 353.34±57.06 µg/l) were significantly lower (p = 0.001) for TB patients when compared with control (550.20±82.33 ng/L and 681.93±97.36µg/l) though within the reference values. Conclusions: This study has shown a lower packed cell volume and haemoglobin concentration in tuberculosis disease indicating the presence of anaemia. An alteration in iron metabolism and increased iron stores has also been demonstrated ruling out iron deficiency anaemia. Vitamin B12 and folate levels though lower for TB patients, are within the reference range thus excluding megaloblastic anaemia. Tuberculosis shows a normocytic normochromic anaemia which is typical of anaemia of chronic disease and inflammation.

Epidemiological profile of intestinal parasites in children under five years of age in Maputo, Mozambique

Intestinal parasites are one of the most sought-after causes in pediatrics. Its occurrence is one of the indicators of socioeconomic status that may be associated with several sociodemographic factors. It is estimated that approximately 50% of the vulnerable population is parasitized with one or more types of parasites, such as children. Considering that this population is essential for human development, the goal of this work was to evaluate the epidemiological profile of intestinal parasites in children up to five years of age in Maputo, Mozambique. To this end, a cross-sectional descriptive study was done with 70 children, through the application of a questionnaire with content related to sociodemographic conditions, clinical profile and identification of the patient’s parasitosis. A stool sample was also requested for parasitological analysis by Ritchie concentration, Ziehl-Neelsen and rapid test techniques. From the 70 children, 36 (51.4%) were infected with at least one intestinal parasite, with Ascaris lumbricoides and Trichuris trichiura being the most prevalent. Furthermore, it was possible to observe a direct correlation between basic sanitation and intestinal parasites. Fever symptoms were also correlated with the presence of intestinal parasitism. Although the government of Mozambique has adopted since 2009 a policy of massive drug administration to deworm children, the high rate of intestinal parasites remains present in the population. Therefore, preventive measures must be taken to contribute to the reduction of this rate of intestinal parasites in this population. KEY WORDS: Public health; Mozambique; kids; intestinal parasites.

Prevalence and Molecular Identification of Cryptosporidium Species among Human Population in Zakho District, Duhok Province, Kurdistan Region, Iraq

This cross-sectional study was carried out to determine the prevalence and molecular identifications of Cryptosporidium species among the human population in Zakho district, Duhok Province, Kurdistan Region, Iraq. The study was performed during the period from August 2021 to March 2022 and involved 700 fecal specimens of both genders and different ages collected from Zakho General Hospital and pri¬mary schools located in the suburban area of Zakho City. The data were analyzed using statistical soft¬ware version SPSS (25). Cryptosporidium species oocyst was found in 87.57% (613/700) of the examined specimens using the Modified Ziehl Neelsen technique. Males showed a slightly higher infection rate than females (88.4% vs 85.5%) with a non-significant difference (P&gt; 0.05). The highest infection rate (91.7%) was present in individuals between the ages of &gt;30 to 40 years, with a statistically significant difference (P&lt;0.05). Molecular identification of Cryptosporidium species was estimated using 18S-rRNA gene for Cryptosporidium genus using Nested PCR technique and genus-specific primers (Cry18 AS1 and Cry18 S2) that yielded 347 bps product indicating Cryptosporidium genus, then species-specific (Cry18 AS1 and Cry18 S1) were used for the detection of Cryptosporidium parvum and Cryptosporidium hominis. 45/50 samples were positive and revealed 240 bps. The acquired sequences were deposited into the GeneBank database under the accession numbers OP107010 for C. hominis, OP104444, OP153870, OP163188, OP163414, and OP164631 for C. parvum.

Screening of Pulmonary Tuberculosis in Household Contacts With Acid Fast Bacteria (AFB) Positive Pulmonary Tuberculosis Patients in Ciwangi Village, Balubur Limbangan Subdistrict, Garut District

Background &amp; Objective Tuberculosis is an infectious disease caused by acid-fast bacteria, namely Mycobacterium tuberculosis, which mainly attacks the lungs. Pulmonary tuberculosis is a global health problem with the second deadliest case after HIV/AIDS infection, and ranks third with the most TB cases after India and China. The main source of transmission is patients with active TB pulmonary tuberculosis who can transmit to people around them. This study aims to determine whether pulmonary TB transmission occurs in people who are in household contact with patients with acid-fast bacteria pulmonary TB active. Method This study is descriptive, using the Ziehl neelsen examination method, with a total sampling technique, conducted on 34 people who are in household contact with pulmonary TB patients in Ciwangi Village, Balubur District, Limbangan. Result Based on the examination results in people who are in contact with people with TB, the results are negative, the presence of the bacteria that cause TB. Conclusion The results of Pulmonary Tuberculosis screening conducted on 34 people from 13 heads of families, it can be concluded that the results of TB microscopy with the Ziehl neelsen technique were negative.

Cryptosporidium infection in diarrheal bovine dairy calves: occurrence and risk factors in Santa Catarina, Brazil

Cryptosporidium protozoa genus are parasites that cause acute enteric disease in young and immunocompromised animals, resulting in anorexia, loss and decrease in weight gain, and, in severe cases, death. Therefore, this study aimed: i) to determine the occurrence of Cryptosporidium spp. in calves with clinical diarrhea in different regions of Santa Catarina, Brazil; ii) to evaluate the risk factors involved with the frequency of infection. iii) to determine the species most involved with the disease in the region. For this, 425 samples were collected in 141 dairy farms, from animals with ages ranging from 0 to 150 days. For this purpose, the samples were submitted to the modified Ziehl-Neelsen technique, with molecular analysis of the positive samples being performed. It was observed 62.1% occurrence of Cryptosporidium spp. in this sampling, especially between 8 to 15 days. Regarding the risk factors evaluated, such as age, management, facilities, water source and Koppen climate (CFA and CFB), none showed statistical significance. Samples positive by the Ziehl-Neelsen technique (32 samples) were randomly selected for molecular diagnosis. Of these, 10 were sequenced, allowing the identification of Crypstosporidium parvum in 6 samples. However, this study proves the existence and high occurrence of the protozoan in different regions of the state of Santa Catarina, Brazil.

Paratuberculose clínica bovina: primeiro relato no estado do Pará

ABSTRACT The objective of this work was to describe the clinical, pathological, and serological aspects of paratuberculosis in a bullock on dairy and beef farm in the municipality of Inhangapí, state of Pará. The animal was raised extensively and presented progressive weight loss, pale mucous membranes, and chronic diarrhea. A blood sample was collected for serology by the indirect ELISA technique. Due to the unfavorable prognosis, the animal was euthanized and necropsied. At necropsy, there was thickened and wrinkled intestinal mucosa, enlarged mesenteric lymph nodes and liquid intestinal contents with a brown color. Organ fragments were fixed in 10% buffered formaldehyde, embedded in paraffin wax, cut, and stained by hematoxylin and eosin and by Ziehl-Neelsen techniques. Histopathology showed intense granulomatous infiltration with Langhans giant cells in the lamina propria and submucosa of the small intestine, Peyer patches and mesenteric lymph nodes. Ziehl-Neelsen staining revealed many intralesional acid-fast bacilli. Antibody searching against Mycobacterium avium (subsp.) paratuberculosis was positive. The diagnosis of paratuberculosis was based on epidemiological, clinical, pathological, and serological data. This is the first report of clinical paratuberculosis, confirmed by pathology and serology, in a bullock in the state of Pará.

Detection rate of pulmonary tuberculosis by direct and concentrated microscopy techniques with Lowenstein-Jensen Culture as standard: South-South Nigeria Scenario

Objective: This study seeks to identify a more sensitive smear microscopy method for the detection of Mycobacterium tuberculosis in low-resource centers. Materials and methods: One hundred and forty sputum specimens were collected and direct smears were prepared as the specimen was submitted. The specimens were afterward digested/decontaminated using the modified Petroff’s method. Both smear methods were stained by the Ziehl-Neelsen technique and examined. All specimens (direct and concentrated) were cultured on a Lowenstein-Jensen medium and results were considered the gold standard to calculate sensitivity. Results: Out of 140 specimens, 27 (19%) and 34 (24%) were positive by direct and concentrated methods respectively. The number of culture positives by the direct and concentrated method was 26 (19%) and 31(22%) respectively. The sensitivity of direct and concentrated methods was 92.3% and 100% respectively. The negative culture and contamination rate was minimum with the concentrated method. The diagnostic accuracy of direct and concentrated methods was 88.6% and 97.1%. Conclusion: In summary, results showed concentrated techniques were able to detect more true positive Mycobacterium tuberculosis than the direct smear method. Therefore, the study region with AFB smear microscopy can utilize the concentrated smear method for AFB smear microscopy to improve the case-finding strategy.

Molecular and phylogenetic characterization of Cryptosporidium species in the saffron finch Sicalis flaveola

BackgroundCryptosporidium is the most common protozoan that can infect a wide variety of animals, including mammals and birds. Fecal samples of six saffron finches, Sicalis flaveola, from a commercial establishment were screened for the presence of Cryptosporidium by the modified Ziehl–Neelsen technique and nested PCR of the 18S rRNA gene followed by sequencing of the amplified fragments.ResultsThe species Cryptosporidium galli was identified in all six saffron fiches, in addition to Cryptosporidium andersoni in one of the birds, indicating a mixed infection. Only two birds had feathers that were ruffled and dirty with feces. Concomitant infection with Isospora spp. was observed in all birds.ConclusionsSaffron finches are a possible host of C. andersoni and this is the first report of this species in a captive bird and the third report of parasitism by C. galli in Sicalis flaveola.

Performance of diagnostic assays used to detect Cryptosporidium oocysts in faecal samples of cattle in Kuwait and genotyping of Cryptosporidium species

BackgroudCryptosporidium species are zoonotic protozoan parasites responsible for gastroenteritis in various animals and humans. The diagnosis of Cryptosporidium presents many challenges. This research attempted to match the diagnostic efficiency of the modified Ziehl–Neelsen technique (mZN), immunochromatographic assays (IC), and enzyme-linked immunosorbent assay (ELISA) for the detection of Cryptosporidium in faecal samples of cattle in Kuwait. In addition, polymerase chain reaction (PCR) was utilised to determine the predominant species infecting cattle in Kuwait and correlating the detected species with the results of different diagnostic tests used, the presence or absence of clinical signs, and the age group of the infected cattle.ResultsOf 400 analysed faecal samples, Cryptosporidium positive samples were 23%, 15.25%, and 14% using IC, ELISA, and mZN. IC had the highest sensitivity (74.07%), and mZN had the highest specificity (98.29%) using a composite reference standard (CRS) as a gold standard. The rapid IC test results in high false-positive results of cryptosporidiosis, whereas using mZN alone is insufficient to declare a negative faecal sample. Only 74.5% (35/47) of Cryptosporidium-positive samples by the three assays could be amplified by PCR. This study was the first to genotype Cryptosporidium in Kuwait. Cryptosporidium parvum (n = 26) was the dominant species detected from cattle samples, followed by C. andersoni (n = 6), C. bovis (n = 2), and C. raynae (n = 1). The findings showed a statistically relevant relationship between diarrhoea and the detection of Cryptosporidium spp. in faecal samples of cattle (p-value = 0.0003). Pre-weaned calves were the most vulnerable age group to Cryptosporidium spp. infection (p-value = 0.0007).ConclusionFor screening of Cryptosporidium infection in faecal samples, antigen detection or PCR methods combined with one of the microscopy techniques should be used. Cryptosporidium parvum was the prepoderant Cryptosporidium spp. recovered from cattle samples in Kuwait followed by C. andersoni. Cryptosporidium parvum is a significant risk factor for diarrhoea in pre-weaned calves. However, further study is needed as many other causes of diarrhoea in calves must be ruled out before a diagnosis of Cryptosporidium diarrhoea can be made.

Prevalence of Cryptosporidium infection in cattle from selected commercial farms and nomadic settlements in Yola, Adamawa State

Cryptosporidium species are apicomplexan parasites commonly associated with diarrhoea in both men and animals. They are of public health importance. The study aimed to determine the occurrence of Cryptosporidium species in cattle in the Yola metropolitan area, Adamawa State, Nigeria. Four hundred and sixteen (416) faecal samples were collected from cattle in commercial farms and nomadic settlements and were analyzed using Modified Ziehl Neelsen (MZN) technique. Twenty-seven (27) positive samples were subjected to nested Polymerase Chain Reaction (PCR) for the amplification of a specific fragment of 18S rRNA gene that was used to detect Cryptosporidium spp. Seventy-three (17.5%) out of 416 samples were positive for Cryptosporidium oocysts by MZN, and 26 (96.0%) out of 27 isolated oocysts of Cryptosporidium detected by MZN were positive by PCR. Commercial farms had a higher prevalence (19.2%) of Cryptosporidium oocysts than the nomadic settlements (15.9%). Based on risk factors, there was a significant association (P&lt;0.05) between the occurrence of Cryptosporidium oocysts and factors such as sex and faecal consistency. At the same time, age, breed, management system, animal source and drinking water source varied insignificantly (P&gt;0.05). This study has shown the occurrence of Cryptosporidium infection in cattle in the Yola metropolis. There should be an awareness campaign among public health workers on the dangers posed by the organism to humans so that the Government would enforce control and preventive measures.

Risk factors for Cryptosporidium infection in small ruminants in northern Greece

The knowledge of risk factors for Cryptosporidium spp. infection in small ruminants is based on limited data. Therefore, the current research aimed to describe the prevalence and risk factors associated with the occurrence of Cryptosporidium infection in sheep and goat herds in northern Greece. Hence, 530 fresh fecal samples from 59 sheep and goat farms were collected and examined for Cryptosporidium oocysts using microscopy of fecal smears stained by the modified Ziehl-Neelsen technique. The overall prevalence of Cryptosporidium infection for both host species was 34% (180/530; 95% confidence interval (CI): 29.9–38). Specifically, the prevalence for sheep and goats was 33.5% (112/334; 95% CI: 28.4–35.6) and 34.7% (68/196; 95% CI: 28–41.4), respectively. Additionally, standardized questionnaires were filled-in to collect data regarding animals’ health status, feeding, and other management practices in each farm. In total 22 risk factors hypothesized to be associated with Cryptosporidium infection were investigated. Multiple logistic regression analysis showed that farms with stagnant water were 11.78 (95% CI: 66–61.5) times more likely to be infected with Cryptosporidium than farms without stagnant water (p < 0.05). Furthermore, farms with more than 25% of their animals suffering from diarrhea were 17.39 (95% CI: 3.43–88.3) times more likely to be infected with Cryptosporidium than farms with ≤ 25% of the animals having diarrhea (p < 0.05). These results suggest that the animal health status and the prevailing environmental conditions play an important role in transmitting Cryptosporidium spp. infection.

Evaluating a Modified Ziehl-Neelsen Technique with Triton X-100 for Detecting Acid-fast Bacilli in Sputum

Evaluating a Modified Ziehl-Neelsen Technique with Triton X-100 for Detecting Acid-fast Bacilli in Sputum

Urinary Mycobacterium tuberculosis Antigens Cocktail as a Potential Marker for Differentiating Active and Latent Tuberculosis Patients

Background: Early secretory antigenic target protein 6-kDa (ESAT-6), culture filtrate 10-kDa (CFP- 10), and Mycobacterium tuberculosis (Mtb) 64 (MPT-64) antigens are secreted by actively growing Mtb in active tuberculosis (TB) patients. Increased levels of ESAT-6, CFP-10, and MPT-64 at the site of infection facilitate the entry of antigens into the systemic circulation. These low molecular weight proteins (&lt; 67 kDa) can be excreted into the urine via the kidneys. Therefore, this study evaluated the urinary Mtb antigen cocktail (ESAT-6, CFP-10, and MPT-64) levels in active [pulmonary (P) and extrapulmonary (EP)] TB and latent tuberculosis infections (LTBI) individuals. Methods: This study was conducted in Dr. Hasan Sadikin General Hospital Bandung from June 2016 to June 2017, using 474 participants. The specimens for TB diagnosis were taken and investigated for acid-fast bacilli (AFB) using the Ziehl Neelsen (ZN) technique. The remaining sputum was cultured on Lowenstein- Jensen (LJ) medium, and TB Ag MPT-64 rapid tests were performed to identify Mtb. In addition, LTBI individuals were screened in a TB outpatient clinic who had close contact with patients with active TB and underwent an interferon g release assay (IGRA). Urine samples were collected for urinary Mtb antigen cocktail (ESAT-6, CFP-10, and MPT-64) testing using the quantitative immunochromatographic method. Results: The 99 patients were divided into three groups, PTB (n = 52), EPTB (n = 22) and LTBI (n = 25). There were significant differences in urinary Mtb antigen cocktail levels between PTB and EPTB (33.8 vs 13.6 ng/ml; p &lt; 0.001), between PTB and LTBI (33.8 vs 0 ng/ml; p &lt; 0.001), and between EPTB and LTBI (13.6 vs 0 ng/ml; p = 0.015). Conclusion: Urinary Mtb antigen cocktail levels were increased in patients with active TB and undetected in those with LTBI. This result was obtained using the urinary Mtb antigen cocktail as a potential marker for differentiating active TB and LTBI.