Multilocus Sequence Typing helps understand the genetic diversity of Cryptosporidium hominis and Cryptosporidium parvum isolated from Colombian patients

Abstract

Multilocus Sequence Typing has become a useful tool for the study of the genetic diversity and population structure of different organisms. In this study, a MLST approach with seven loci (CP47, MS5, MS9, MSC6-7, TP14, and gp60) was used to analyze the genetic diversity of Cryptosporidium hominis and Cryptosporidium parvum isolated from 28 Colombian patients. Five Cryptosporidium species were identified: C. hominis, C. parvum, Cryptosporidium felis, Cryptosporidium meleagridis, and Cryptosporidium suis. Unilocus gp60 analysis identified four allelic families for C. hominis (Ia, Ib, Id, and Ie) and two for C. parvum (IIa and IIc). There was polymorphic behavior of all markers evaluated for both C. hominis and C. parvum, particularly with the CP47, MS5, and gp60 markers. Phylogenetic analysis with consensus sequences (CS) of the markers showed a taxonomic agreement with the results obtained with the 18S rRNA and gp60 gene. Additionally, two monophyletic clades that clustered the species C. hominis and C. parvum were detected, with a higher number of subclades within the monophyletic groups compared to those with the gp60 gene. Thirteen MLG were identified for C. hominis and eight for C. parvum. Haplotypic and nucleotide diversity were detected, but only the latter was affected by the gp60 exclusion from the CS analysis. The gene fixation index showed an evolutionary closeness between the C. hominis samples and a less evolutionary closeness and greater sequence divergence in the C. parvum samples. Data obtained in this work support the implementation of MLST analysis in the study of the genetic diversity of Cryptosporidium, considering the more detailed information that it provides, which may explain some genetic events that with an unilocus approach could not be established. This is the first multilocus analysis of the intra-specific variability of Cryptosporidium from humans in South America.

Bioprospecting of Aspergillus sp. as a promising repository for anti-cancer agents: a comprehensive bibliometric investigation

“…This method plays a crucial role in mapping genetic relationships and variations among Aspergillus strains, providing insights essential for both fundamental research and practical applications. Furthermore, Multilocus Sequence Typing (MLST) is used to gain a comprehensive view of the genetic landscape by analyzing multiple genetic loci, which is pivotal for understanding the evolutionary paths and relationships among different fungal strains ( Uran-Velasquez et al, 2022 ).…”

Cryptosporidium and Giardia in cats and dogs: What is the real zoonotic risk?

“…Although C. felis , C. canis , and assemblages C, D and F are considered to be of limited zoonotic potential, this risk is not negligible as there have been numerous reports of C. felis and C. canis in humans, as well as a few reports of assemblages C, D and F in humans ( Table 3 ). The prevalence of C. felis and C. canis in humans is usually low (1–3% of cryptosporidiosis cases); however, a few studies have reported higher prevalences; 5–40% for C. canis and 7.1–37.5% for C. felis cryptosporidiosis cases, mostly in children or HIV-positive individuals ( Moore et al, 2016 ; Dacal et al, 2018 ; Sannella et al, 2019 ; Uran-Velasquez et al, 2022 ; Wang et al, 2022b ; Khalifa et al, 2023 ).…”

“… Species/Assemblage No. of cases Comment Country Reference Cryptosporidium C. felis 2 HIV+ patients Algeria Semmani et al (2023) 1 21-year-old female with diarrhoea Australia Ebner et al (2015) 5 HIV+ patients Brazil Lucca et al (2009) 4 School children Brazil Pacheco et al (2022) 2 Sporadic cases Canada Guy et al (2021) 2 Hospitalised children (median 36 months-old) China Feng et al (2012) 8 1 HIV+, 4 with diarrhoea, 2 with non-gastrointestinal illnesses China Liu et al (2020) 3 Kindergarten children aged 2–6 years China Wang et al (2022b) 1 Average age 5 years Colombia Higuera et al (2020) 2 1 child and 1 HIV+ patient Colombia Uran-Velasquez et al (2022) 1 C. felis identified in owner’s cat Colombia Potes-Morales and Crespo-Ortiz (2023) ...…”

WHOLE GENOME TARGETED ENRICHMENT AND SEQUENCING OF HUMAN-INFECTINGCRYPTOSPORIDIUMspp.

“…Several molecular methods have been developed to gain insight into the genetic diversity of Cryptosporidium species, often relying on single marker typing of conserved (small subunit rRNA, SSU) or polymorphic regions (glycoprotein gene 60, gp60, or heat shock protein 70, hsp70) 15,28 . Other approaches for genetic characterization include multilocus sequence typing (MLST) [29][30][31][32] , and more recently, whole genome sequencing. The disadvantages of MLST include the possible unreliability of markers in different regions due to the limited information about the genetic variability of populations within Cryptosporidium species 32 , and thus, assays are not universal.…”

“…Other approaches for genetic characterization include multilocus sequence typing (MLST) [29][30][31][32] , and more recently, whole genome sequencing. The disadvantages of MLST include the possible unreliability of markers in different regions due to the limited information about the genetic variability of populations within Cryptosporidium species 32 , and thus, assays are not universal. Challenges with comprehensive genomic assays such as whole genome sequencing are constrained by difficulty in purifying oocysts 33 and the limited amount of DNA from purified samples and from fecal clinical samples (which also have an abundance of other organisms' DNA).…”