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. 2019 Jan;12(1):183-189.
doi: 10.14202/vetworld.2019.183-189. Epub 2019 Jan 31.

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

Kaltoum Yagoub Adam  1 , A A Ismail  2 , M A Masri  3 , A A Gameel  4
Affiliations

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

Kaltoum Yagoub Adam et al. Vet World. 2019 Jan.

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.

Keywords: Cryptosporidium parvum; Sudan; nested polymerase chain reaction; staining techniques; zoonotic.

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Figures

Figure-1
Figure-1
Representative ethidium bromide stained agarose gel showing polymerase chain reaction (PCR) products of Cryptosporidium DNA extracts from fecal samples. M: Molecular marker, Lanes: 2-11 nested PCR products.
Figure-2
Figure-2
Phylogenetic relationships of the isolates examined in the current study (labeled) to different Cryptosporidium spp. as inferred by Neighbor-joining analysis of the 18s rRNA gene.
Figure-3
Figure-3
Alignment of Cryptosporidium parvum current isolates (KG1 and KG2) with selected C. parvum reference sequences showing different positions.
See this image and copyright information in PMC

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