Cryptosporidium oocyst shedding in buffalo calves in Haryana: A case study

2018 ◽  
Author(s):  
Krutanjali Swain ◽  
Abhilash Routray ◽  
Saraswat Sahoo ◽  
Subha Ganguly
Keyword(s):  
Cryptosporidium Oocyst ◽  
Buffalo Calves ◽  
Concentration Technique ◽  
Cryptosporidium Oocysts ◽  
Oil Immersion ◽  
Neonatal Diarrhoea ◽  
Conventional Microscopy ◽  
Animal Sciences ◽  
Acid Fast Staining

Bovine cryptosporidiosisis primarily associated with neonatal diarrhoea with higher morbidity than mortality in young calves till they attain immunological maturity. The present investigation relates to a report on the shedding of Cryptosporidium oocyst in two buffalo calves of buffalo farm, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar from 1st days up to 3 month of age at 15 days interval using simple conventional microscopy. By using formol-ether concentration technique followed by modified Ziehl-Neelsen (ZN) acid fast staining, Cryptosporidium oocysts were concentrated and identified. The Cryptosporidium oocysts appeared as reddish pink coloured bodies against a bluish/greenish coloured background at oil immersion using ZN staining kit. The maximum oocyst shedding was observed (2.3 oocyst / field) during 16 to 30 days of age. The oocyst shedding gradually decreased with increase in age afterwards clearly indicating the disease of young buffalo calves.

Cryptosporidium oocyst shedding in buffalo calves in Haryana: A case study

2018 ◽  
Author(s):  
Krutanjali Swain ◽  
Abhilash Routray ◽  
Saraswat Sahoo and Subha Ganguly
Keyword(s):  
Cryptosporidium Oocyst ◽  
Buffalo Calves ◽  
Concentration Technique ◽  
Cryptosporidium Oocysts ◽  
Oil Immersion ◽  
Neonatal Diarrhoea ◽  
Conventional Microscopy ◽  
Animal Sciences ◽  
Acid Fast Staining

Bovine cryptosporidiosisis primarily associated with neonatal diarrhoea with higher morbidity than mortality in young calves till they attain immunological maturity. The present investigation relates to a report on the shedding of Cryptosporidium oocyst in two buffalo calves of buffalo farm, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar from 1st days up to 3 month of age at 15 days interval using simple conventional microscopy. By using formol-ether concentration technique followed by modified Ziehl-Neelsen (ZN) acid fast staining, Cryptosporidium oocysts were concentrated and identified. The Cryptosporidium oocysts appeared as reddish pink coloured bodies against a bluish/greenish coloured background at oil immersion using ZN staining kit. The maximum oocyst shedding was observed (2.3 oocyst / field) during 16 to 30 days of age. The oocyst shedding gradually decreased with increase in age afterwards clearly indicating the disease of young buffalo calves.

scholarly journals Use of Oxidative Stress Responses to Determine the Efficacy of Inactivation Treatments on Cryptosporidium Oocysts

2021 ◽  
Vol 9 (7) ◽  
pp. 1463
Author(s):  
Tamirat Tefera Temesgen ◽  
Kristoffer Relling Tysnes ◽  
Lucy Jane Robertson
Keyword(s):  
Oxidative Stress ◽  
Stress Responses ◽  
Target Genes ◽  
Cryptosporidium Oocyst ◽  
Drinking Water Treatment ◽  
Proof Of Concept ◽  
Environmental Matrices ◽  
Oocyst Wall ◽  
Novel Approach ◽  
Cryptosporidium Oocysts

Cryptosporidium oocysts are known for being very robust, and their prolonged survival in the environment has resulted in outbreaks of cryptosporidiosis associated with the consumption of contaminated water or food. Although inactivation methods used for drinking water treatment, such as UV irradiation, can inactivate Cryptosporidium oocysts, they are not necessarily suitable for use with other environmental matrices, such as food. In order to identify alternative ways to inactivate Cryptosporidium oocysts, improved methods for viability assessment are needed. Here we describe a proof of concept for a novel approach for determining how effective inactivation treatments are at killing pathogens, such as the parasite Cryptosporidium. RNA sequencing was used to identify potential up-regulated target genes induced by oxidative stress, and a reverse transcription quantitative PCR (RT-qPCR) protocol was developed to assess their up-regulation following exposure to different induction treatments. Accordingly, RT-qPCR protocols targeting thioredoxin and Cryptosporidium oocyst wall protein 7 (COWP7) genes were evaluated on mixtures of viable and inactivated oocysts, and on oocysts subjected to various potential inactivation treatments such as freezing and chlorination. The results from the present proof-of-concept experiments indicate that this could be a useful tool in efforts towards assessing potential technologies for inactivating Cryptosporidium in different environmental matrices. Furthermore, this approach could also be used for similar investigations with other pathogens.

scholarly journals Multicenter Evaluation of an ELISA for the Detection of Cryptosporidium spp. Antigen in Clinical Human Stool Samples

2021 ◽  
Vol 9 (2) ◽  
pp. 209
Author(s):  
Romy Razakandrainibe ◽  
Célia Mérat ◽  
Nathalie Kapel ◽  
Marc Sautour ◽  
Karine Guyot ◽  
...  
Keyword(s):  
Large Scale ◽  
Cryptosporidium Oocyst ◽  
Clinical Importance ◽  
Epidemiological Studies ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cryptosporidium Spp ◽  
Stool Samples ◽  
Conventional Microscopy ◽  
Human Stool

Human cryptosporidiosis remains underdiagnosed, and rapid/accurate diagnosis is of clinical importance. Diagnosis of the Cryptosporidium oocyst in stool samples by conventional microscopy is labor-intensive, time-consuming, and requires skillful experience. Thus, we aimed to evaluate the usefulness of a coproantigen enzyme-linked immunosorbent assay (ELISA) test in detecting Cryptosporidium spp. from fecal specimens. For this aim, we evaluated the performances of a commercial ELISA (CoproELISA Cryptosporidium kit, Savyon Diagnostics, Israel) for the detection of Cryptosporidium spp. in random clinical stool samples through a multicenter study. The sensitivity and specificity for coproantigen ELISA were 98.86% and 94.32%, respectively. The coproantigen ELISA results indicate that the simple, rapid, reliable, and standardized immunoassay test is sensitive and specific for routine diagnosis, and may be useful for large-scale epidemiological studies of cryptosporidiosis.

Determination of the Recovery Efficiency of Cryptosporidium Oocysts and Giardia Cysts from Seeded Bivalve Mollusks

2013 ◽  
Vol 76 (1) ◽  
pp. 93-98 ◽  
Author(s):  
FRANCISKA M. SCHETS ◽  
HAROLD H. J. L. van den BERG ◽  
ANA MARIA de RODA HUSMAN
Keyword(s):  
Quantitative Assessment ◽  
Intestinal Parasites ◽  
Cryptosporidium Oocyst ◽  
Immunomagnetic Separation ◽  
Giardia Cyst ◽  
Bivalve Mollusks ◽  
Recovery Efficiency ◽  
Risk Of Infection ◽  
Cryptosporidium Oocysts

The intestinal parasites Cryptosporidium and Giardia are transmitted by water and food and cause human gastroenteritis. Filter-feeding bivalve mollusks, such as oysters and mussels, filter large volumes of water and thus concentrate such pathogens, which makes these bivalves potential vectors of disease. To assess the risk of infection from consumption of contaminated bivalves, parasite numbers and parasite recovery data are required. A modified immunomagnetic separation (IMS) procedure was used to determine Cryptosporidium oocyst and Giardia cyst numbers in individually homogenized oysters (Crassostrea gigas) and mussels (Mytilus edulis). About 12% of the commercial bivalves were positive, with low (oo)cyst numbers per specimen. The recovery efficiency of the IMS procedure was systematically evaluated. Experiments included seeding of homogenized bivalves and whole animals with 100 to 1,000 (oo)cysts. Both seeding procedures yielded highly variable recovery rates. Median Cryptosporidium recoveries were 7.9 to 21% in oysters and 62% in mussels. Median Giardia recoveries were 10 to 25% in oysters and 110% in mussels. Giardia recovery was significantly higher than Cryptosporidium recovery. (Oo)cysts were less efficiently recovered from seeded whole animals than from seeded homogenates, with median Cryptosporidium recoveries of 5.3% in oysters and 45% in mussels and median Giardia recoveries of 4.0% in oysters and 82% in mussels. Both bivalve homogenate seeding and whole animal seeding yielded higher (oo)cyst recovery in mussels than in oysters, likely because of the presence of less shellfish tissue in IMS when analyzing the smaller mussels compared with the larger oysters, resulting in more efficient (oo)cyst extraction. The data generated in this study may be used in the quantitative assessment of the risk of infection with Cryptosporidium or Giardia associated with the consumption of raw bivalve mollusks. This information may be used for making risk management decisions.

scholarly journals Cryptosporidium infection in non-human hosts in Malawi

2009 ◽  
Vol 76 (4) ◽  
Author(s):  
Z. Banda ◽  
Rosely A.B. Nichols ◽  
A.M. Grimason ◽  
H.V. Smith
Keyword(s):  
Cryptosporidium Parvum ◽  
Rainy Season ◽  
18S Rrna ◽  
Cryptosporidium Oocyst ◽  
Dry Season ◽  
Cool Season ◽  
Cryptosporidium Hominis ◽  
Adult Cattle ◽  
Faecal Samples ◽  
Cryptosporidium Oocysts

Of 1 346 faecal samples from the Chikwawa and Thyolo districts of Malawi, analysed for the presence of Cryptosporidium oocysts between October 2001 and May 2003, 61.3 % were from cattle (29.8 % of these were from calves < 6 months old). Cryptosporidium oocysts were detected during all three seasons studied in Chikwawa and Thyolo. In Chikwawa, 13.6 % of adult cattle and 11.7 % of calves were infected, compared to 28.9 % of adult cattle and 36.7 % of calves in Thyolo. Dependent on season, between 7.8 % and 37.7 % (Chikwawa) and 16.7 % and 39.3 % (Thyolo) of cattle samples contained oocysts. In Chikwawa, the highest percentage of infections occurred in the cool season, whereas in Thyolo, the highest percentage of infections occurred in the dry season. Faecal samples from goats [n = 225], pigs [n = 92], sheep [n = 6]), rabbits, guinea pigs, chickens, ducks, turkeys, doves and guinea fowls were also analysed. Up to 5.6 % of goat samples contained oocysts in Chikwawa, compared to between 16.7 % and 39.3 % in Thyolo. Again, in Chikwawa, the highest percentage of infections occurred in the cool season and the lowest in the rainy season, whereas, in Thyolo, the highest percentage of infections occurred in the dry season and the lowest in the cool season. In pigs, more infections were detected in the dry season in Chikwawa, but infections in the cool season were similar (17.7 %), whereas in Thyolo, infections occurred in all three seasons (17.9 % in the rainy season, 25 % in the cool season and 60 % in the dry season). Often diarrhoeic, oocyst positive cattle faecal samples collected from Chikwawa and subjected to PCR-RFLP, four oocyst positive samples (two from heifers, one from a cow and one unknown) were amplified at an 18S rRNA and Cryptosporidium oocyst wall protein (COWP) loci. RFLP of the 18S rRNA locus indicated that Cryptosporidium parvum, Cryptosporidium hominis, Cryptosporidium bovis and / or Cryptosporidium ryanae DNA, or a mixture of them was present. Cryptosporidium parvum DNA was identified in one sample that amplified at the COWP locus, indicating the presence of the major zoonotic Cryptosporidium species in Malawi.

scholarly journals Microrheology imaging of fiber suspensions – a case study for lyophilized collagen I in HCl solutions

Soft Matter ◽  
2020 ◽  
Vol 16 (39) ◽  
pp. 9014-9027
Author(s):  
Johanna Hafner ◽  
Claude Oelschlaeger ◽  
Norbert Willenbacher
Keyword(s):  
Network Structure ◽  
Particle Tracking ◽  
Collagen I ◽  
Fiber Suspensions ◽  
Direct Visualization ◽  
Multiple Particle Tracking ◽  
Conventional Microscopy ◽  
Multiple Particle

Where conventional microscopy fails, overlaying subsequent images of multiple particle tracking (MPT) videos including short trajectories allowed for direct visualization of the network structure of lyophilized collagen I.

scholarly journals Cryptosporidium spp. Infections in Livestock and Wild Animals in Azerbaijan Territory

2020 ◽  
Vol 2 (1) ◽  
pp. 44
Author(s):  
Simuzer Mamedova ◽  
Panagiotis Karanis
Keyword(s):  
Staining Method ◽  
Current Knowledge ◽  
Protozoan Parasite ◽  
Structural Features ◽  
Cryptosporidium Spp ◽  
Faecal Samples ◽  
Cryptosporidium Oocysts ◽  
Stool Specimens ◽  
Intracellular Protozoan Parasite ◽  
Acid Fast Staining

Cryptosporidium is an intracellular protozoan parasite and is increasingly gaining attention as a human and an animal pathogen, mainly due to its predominant involvement in worldwide waterborne outbreaks. This paper reviews the current knowledge and understanding of Cryptosporidium spp. in terrestrial and aquatic animals in Azerbaijan. The diagnosis of cryptosporidiosis relies on the identification of oocysts in faecal samples released by the infected host. Stool specimens were processed using the modified acid-fast staining method (Ziehl-Neelsen) and microscopically examined for Cryptosporidium oocysts. Thirteen species of Cryptosporidium (C. fragile, C. ducismarci, C. serpentis, C. varani, C. baileyi, C. meleagridis, C. muris, C. parvum, C. ubiquitum, C. andersoni, C. bovis, C. hominis, C. suis) from amphibians, reptiles, birds and mammals have been identified as a result of studies conducted between 1987 and 2019 on the structural features of Cryptosporidium oocysts in Azerbaijan territory.

Microwave Inactivation of Cyclospora cayetanensis Sporulation and Viability of Cryptosporidium parvum Oocysts

2006 ◽  
Vol 69 (8) ◽  
pp. 1957-1960 ◽  
Author(s):  
YNES R. ORTEGA ◽  
JYEYIN LIAO
Keyword(s):  
Cryptosporidium Parvum ◽  
Propidium Iodide ◽  
Cryptosporidium Oocyst ◽  
Cooking Time ◽  
Cyclospora Cayetanensis ◽  
The Third ◽  
Infectivity Assay ◽  
Cryptosporidium Oocysts ◽  
Cryptosporidium Parvum Oocysts ◽  
Cooking Times

The efficacy of microwave heating on the viability of Cryptosporidium parvum oocysts and on the sporulation of Cyclospora cayetanensis oocysts for various periods of cooking times (0, 10, 15, 20, 30, and 45 s) at 100% power was determined. Cyclospora oocysts were stored in 2.5% dichromate at 23°C for 2 weeks, and sporulation rates were then determined. The 4′,6-diamidino-2-phenylindole and propidium iodide vital stain and the neonate animal infectivity assay determined Cryptosporidium oocyst viability. Cryptosporidium oocysts could be completely inactivated with as little as 20 s of cooking time, whereas Cyclospora sporulation was observed up to 45 s. Two of the examined microwave ovens were more effective at reducing sporulation and viability than the third one. Because of the variability of temperature achieved by the various ovens, cooking time was not an accurate parameter for parasite inactivation. Cryptosporidium oocysts could be inactivated only when temperatures of 80°C or higher were reached in the microwave ovens.

scholarly journals First report of Cryptosporidium spp. oocysts in oysters (Crassostrea rhizophorae) and cockles (Tivela mactroides) in Brazil

2008 ◽  
Vol 6 (4) ◽  
pp. 527-532 ◽  
Author(s):  
Diego Averaldo Guiguet Leal ◽  
Mirna Aparecida Pereira ◽  
Regina Maura Bueno Franco ◽  
Nilson Branco ◽  
RomeuCantusio Neto
Keyword(s):  
Hepatitis A Virus ◽  
Cryptosporidium Oocyst ◽  
Coastal Region ◽  
Tween 80 ◽  
Public Health Importance ◽  
First Report ◽  
Cryptosporidium Oocysts ◽  
Thermotolerant Coliforms ◽  
Seawater Quality ◽  
Crassostrea Rhizophorae

The consumption of oysters and cockles, which are usually eaten raw or lightly-cooked, can cause outbreaks of human diseases, especially if these shellfish are harvested from polluted areas. In Brazil data about the occurrence of pathogens, like hepatitis A virus, in shellfish have been reported but research on natural contamination for pathogenic protozoa is still non-existent. Cryptosporidium oocyst contamination of oysters (Crassostrea rhizophorae) and cockles (Tivela mactroides) was evaluated during two different periods in a coastal area from São Paulo, Brazil. From June to November 2005, and from July to December 2006, 180 mollusks were harvested for tissue examination. The gills and gastrointestinal tract (n = 36 pools) were carefully extracted from the animals and homogenized in a tissue homogenizer by adding surfactant Tween 80 (0.1%). Immunofluorescence assays were performed and Cryptosporidium oocysts were detected in 50.0% of gill pools of cockles and 10.0% of gill pools of oysters. In order to evaluate seawater quality in shellfish growing areas, total levels of thermotolerant coliforms, Escherichia coli and enterococci were determined. This is the first time that Cryptosporidium oocysts were found in shellfish from the coastal region of Brazil, and to the best of our knowledge it is also the first report in Latin America and the case might be of public health importance, reflecting the extension of the contamination on seafood, requiring a need for quality control standards.

On-Chip Trapping and Characterization of Cryptosporidium Using Surface Coated ITO-PDMS Bonded Chips

2011 ◽  
Vol 254 ◽  
pp. 191-194
Author(s):  
Harikrishnan Narayanan Unni ◽  
Deny Hartono ◽  
Kian Meng Lim
Keyword(s):  
Cryptosporidium Oocyst ◽  
Rapid Identification ◽  
Channel Height ◽  
Cell Trapping ◽  
Cell Compartments ◽  
Silane Coating ◽  
Ito Glass ◽  
Cryptosporidium Oocysts ◽  
On Chip

Dielectrophoresis has been shown to have significant potential for the characterization of cells and could become an efficient tool for rapid identification and assessment of microorganisms. The present work is focused on the trapping and characterization of Cryptosporidium pathogen using a microfluidic chip fabricated through a simple, effective bonding of surface coated ITO glass and PDMS. Lithographically patterned ITO glass plates were coated with an alkoxysilane solution DMOAP (N, N – dimethyl -N- octadecyl-3-aminopropyltrimethoxysilyl chloride) prior to bonding with PDMS microchannels. The silane coating was found to enhance the bonding between ITO and PDMS and reduce cell adhesion on the electrodes. Cryptosporidium oocysts, which are 2-4 microns in size and nearly spherical in shape represent the preliminary stage of cell development. The dielectrophoretic transport of cells is dependent on electrical properties such as permittivity and conductivity of the cells. Computational simulations were performed in order to study the effects of channel height, buffer conductivity and applied voltage on the flow and cell transport inside the DEP chip and facilitate effective cell trapping. Videomicroscopic experiments were performed using the fabricated device and the real part of Clausius-Mossotti factor of the cells was estimated from values of critical voltages of particle trapping (corresponding to various field frequencies) at the electrodes. The dielectric properties of the cell compartments (cytoplasm and membrane) were calculated based on a single shell model of the Cryptosporidium oocyst cell.

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