Abstract
Background and Aim:Cryptosporidium is recognized to infect several mammalian species as well as humans, causing substantial economic losses and serious public health concern. Infected animals can be a source of environmental contamination and human infections. In general, the occurrence of Cryptosporidium species in animals and human in Sudan and zoonotic importance is not well documented. This study aimed to identify Cryptosporidium spp. infecting different animal species and humans and to compare between different isolates obtained.Materials and Methods:To provide molecular information about Cryptosporidium in animals and humans, both modified Ziehl–Neelsen (MZN) specific stain and molecular assay were used. Concentration techniques followed by three protocols of DNA extraction were carried out. After microscopic screening of 263 fecal samples (goats [n=197], cattle [n=12], sheep [n=12], and human [n=42]), 61 positive and 30 negative, randomly selected samples were used in nested polymerase chain reaction (PCR) targeting part of the 18S RNA.Results:Nested PCR amplification confirmed 91.8% (56/61) of microscopic-positive samples. 8.2% (5/61) of negative samples by PCR (positive by microscopy) were considered false negatives. Sequencing followed by alignment of the 14 isolates indicated that all samples were identical (100%) and belonged to Cryptosporidium parvum.Conclusion:MZN staining procedure is reliable for the routine diagnosis of Cryptosporidium; cetyltrimethylammonium bromide extraction buffer and nested PCR targeting 18S rRNA gene are reliable and useful in epidemiological studies of this parasite.
Highlights
The genus Cryptosporidium consists of protozoan parasites that invade the microvillus border of the gastrointestinal epithelium of all classes of vertebrates
modified Ziehl–Neelsen (MZN) staining procedure is reliable for the routine diagnosis of Cryptosporidium; cetyltrimethylammonium bromide extraction buffer and nested polymerase chain reaction (PCR) targeting 18S rRNA gene are reliable and useful in epidemiological studies of this parasite
Three methods of DNA extraction were used in the present study, only cetyltrimethylammonium bromide (CTAB) and phenol methods were successful in producing PCR products in subsequent PCR reaction
Summary
The genus Cryptosporidium consists of protozoan parasites that invade the microvillus border of the gastrointestinal epithelium of all classes of vertebrates. Cryptosporidium spp. infects a wide range of livestock animals and humans [2,3,4,5,6,7,8]. Those parasites are ubiquitous in the environment, and oocysts are extremely resistant to environmental conditions and many of the commonly used disinfectants [9]. Infected animals can be a source of environmental contamination and human infections. The occurrence of Cryptosporidium species in animals and human in Sudan and zoonotic importance is not well documented. This study aimed to identify Cryptosporidium spp. infecting different animal species and humans and to compare between different isolates obtained
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