Long-Term Rasamsonia argillacea Complex Species Colonization Revealed by PCR Amplification of Repetitive DNA Sequences in Cystic Fibrosis Patients.

The aim of this work was to document molecular epidemiology of Rasamsonia argillacea species complex isolates from cystic fibrosis (CF) patients. In this work, 116 isolates belonging to this species complex and collected from 26 CF patients and one patient with chronic granulomatous disease were characterized using PCR amplification assays of repetitive DNA sequences and electrophoretic separation of amplicons (rep-PCR). Data revealed a clustering consistent with molecular species identification. A single species was recovered from most patients. Rasamsonia aegroticola was the most common species, followed by R. argillacea sensu stricto and R. piperina, while R. eburnea was not identified. Of 29 genotypes, 7 were shared by distinct patients while 22 were patient specific. In each clinical sample, most isolates exhibited an identical genotype. Genotyping of isolates recovered from sequential samples from the same patient confirmed the capability of R. aegroticola and R. argillacea isolates to chronically colonize the airways. A unique genotype was recovered from two siblings during a 6-month period. In the other cases, a largely dominant genotype was detected. Present results which support the use of rep-PCR for both identification and genotyping for the R. argillacea species complex provide the first molecular evidence of chronic airway colonization by these fungi in CF patients.

Semi-automated repetitive sequence-based PCR amplification for species of the Scedosporium apiospermum complex.

PURPOSE: The Scedosporium apiospermum species complex usually ranks second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF), but little is known about the molecular epidemiology of the airway colonization. METHODS: Polymerase chain reaction (PCR) amplification of repetitive sequences (rep-PCR) was applied to the retrospective analysis of a panel of isolates already studied by random amplification of polymorphic DNA (RAPD) and comprising 63 isolates recovered from sputa from 9 CF patients. Results were compared to those obtained previously by RAPD, and herein by beta-tubulin (TUB) gene sequencing and Multilocus Sequence Typing (MLST). RESULTS: Within the panel of isolates studied,S. apiospermum sensu stricto and Scedosporium boydii, as expected, were the predominant species with 21 and 36 isolates, respectively. Four isolates from one patient were identified as Scedosporium aurantiacum, whereas two isolates belonged to the Pseudallescheria ellipsoidea subgroup of S. boydii rep-PCR analysis of these isolates clearly differentiated the three species and P. ellipsoidea isolates, whatever the rep-PCR kit used, and also permitted strain differentiation. When using the mold primer kit, results from rep-PCR were in close agreement with those obtained by MLST. For both S. apiospermum and S. boydii, 8 genotypes were differentiated by rep-PCR and MLST compared to 10 by RAPD. All S. aurantiacum isolates shared the same RAPD genotype and exhibited the same rep-PCR profile and sequence type. CONCLUSIONS: These results illustrate the efficacy of rep-PCR for both species identification within the S. apiospermum complex and genotyping for the two major species of this complex.Abstract presentation: Part of this work was presented during the 18th Congress of the International Society for Human and Animal Mycology, Berlin (Germany), June 2012.S. Giraud, C. Godon, A. Rougeron, J.P. Bouchara and L. Favennec are members of the ECMM/ISHAM working group on Fungal respiratory infections in Cystic Fibrosis(Fri-CF).

Occurrence and Molecular Characterization of Cryptosporidium Infection in HIV/Aids Patients in Algeria.

The estimated prevalence rate of adults living with HIV infection in MENA is one of the lowest in the world. To date, no data on the genetic characteristics of Cryptosporidium isolates from HIV/AIDS patients in Algeria were available. This study aimed to identify Cryptosporidium species and subtype families prevalent in Algerian HIV-infected patients and contribute to the molecular epidemiology mapping of Cryptosporidium in the MENA region. A total of 350 faecal specimens from HIV/AIDS patients were analysed using microscopy, and a Cryptosporidium infection was identified from 33 samples, with 22 isolates successfully sequencing and confirming species and subtypes. Based on sequence analysis, 15 isolates were identified as C. parvum with family subtypes IIa (n = 7) and IId (n = 8), while five were identified as C. hominis (family subtypes Ia (n = 2) and Ib (n = 3)) and two as C. felis. The C. parvum subtype families IIa and IId predominated, suggesting potential zoonotic transmission. More extensive sampling of both humans and farm animals, especially sheep, goats and calves, as well as a collection of epidemiological data are needed for a better understanding of the sources of human C. parvum infections in Algeria.

Genetic diversity of Cryptosporidium spp. in non-human primates in rural and urban areas of Ethiopia.

Non-Human Primates (NHPs) harbor Cryptosporidium genotypes that can infect humans and vice versa. NHPs Chlorocebus aethiops and Colobus guereza and humans have overlapping territories in some regions of Ethiopia, which may increase the risk of zoonotic transmission of Cryptosporidium. This cross-sectional study examined the molecular prevalence and subtypes of Cryptosporidium spp. from 185 fecal samples of Chlorocebus aethiops and Colobus guereza in rural and urban areas in Ethiopia. Samples were tested for Cryptosporidium infection using nested polymerase chain reaction (PCR), and subtypes were determined by sequencing a fragment of the 60-kDa glycoprotein gene (gp60). Of the 185 samples, fifty-one (27.56%) tested positive for Cryptosporidium infection. The species detected were C. parvum (n = 34), C. hominis (n = 12), and C. cuniculus (n = 3). Mixed infection with C. parvum and C. hominis were detected in 2 samples. Four C. hominis family subtypes (Ia, Ib, Id, and Ie) and one C. parvum family subtype (IIa) were identified. C. hominis IaA20 (n = 7) and C. parvum IIaA17G1R1 (n = 6) were the most prevalent subtypes detected. These results confirm that Chlorocebus aethiops and Colobus guereza can be infected with diverse C. parvum and C. hominis subtypes that can also potentially infect humans. Additional studies could help to understand the role of NHPs in the zoonotic transmission of Cryptosporidium in Ethiopia.

A summary of cryptosporidiosis outbreaks reported in France and overseas departments, 2017-2020.

Cryptosporidium is a known foodborne pathogen, ranked fifth out of 24 among foodborne parasites in terms of importance and a cause of many cryptosporidiosis outbreaks worldwide. In France, very few outbreaks were reported before 2017, and data recently obtained by the Expert Laboratory of the Cryptosporidiosis National Reference Center (CNR-LE-Cryptosporidiosis) have shown that outbreaks are in fact common and frequently underreported. In this work, we aim to report the characteristics of outbreaks detected in France during the period 2017-2020 and present a summary of investigations carried out by the CNR-LE-Cryptosporidiosis. During the study period, there were eleven cryptosporidiosis outbreaks, including three with no identified origin. Among the eight identified outbreaks: six were due to water contamination (five tap water and one recreational water), one was due to direct contact with infected calves, and one was due to consumption of contaminated curd cheese. Among these outbreaks, five of them exceeded one hundred cases. Recent results obtained by the CNR-LE-Cryptosporidiosis revealed the multiannual occurrence of Cryptosporidium outbreaks in France. Waterborne outbreaks were more frequently detected, while foodborne outbreaks which are more difficult to detect were likely underreported.

Molecular characterization of Cryptosporidium spp. from humans in Ethiopia.

Data on the distribution and genotype of Cryptosporidium species is limited in Ethiopia. This study examined the presence and genetic diversity of Cryptosporidium species circulating in Ethiopian human population. Stool samples collected from patients who visited rural (n = 94) and urban (n = 93) health centers in Wurgissa and Hawassa district, respectively, were examined for the presence of Cryptosporidium spp. using microscopy, nested PCR and real-time PCR. To detect infection with PCR, analysis of 18S ribosomal RNA was performed. Subtyping was performed by sequencing a fragment of GP60 gene. The overall prevalence of infection was 46% (n = 86) by microscope and PCR. When 48 (out of 86) PCR positive samples were genotyped, two species were identified: C. parvum (n = 40) and C. hominis (n = 8). When 15 of the 40 C. parvum isolates were subtyped, zoonotic subtypes of IIaA14G1R1 (n = 1), IIaA15G2R1 (n = 1), IIaA16G1R1 (n = 2), IIaA16G3R1 (n = 2), IIaA17G1R1 (n = 1), IIaA19G1R1 (n = 1), IIaA20G1R1 (n = 3), IIaA22G1R1 (n = 1), IIaA22G2R1 (n = 1), IIdA23G1 (n = 1) and IIdA24G1 (n = 1) were identified. When 6 of the 8 C. hominis isolates were subtyped, subtypes IaA20 (n = 5), and IdA21(n = 1) were identified. This study suggests that C. parvum and C. hominis are causes of cryptosporidiosis in human in the Wurgissa district and Hawassa in Ethiopia. Zoonotic transmission might be the main route of transmission.

Scedosporium species in soils from various biomes in Northwestern Morocco.

Scedosporium species are opportunistic pathogens causing various infections, including disseminated infections in severely immunocompromised patients. Preventive measures aiming to reduce the risk of exposure to these fungi require a better knowledge on their ecology and on the sources of contamination of the patients. In this context, 99 soil samples from the Rabat-Sale-Kenitra and Fez-Meknes regions in Morocco were analyzed. Samples were inoculated on the highly selective Scedo-Select III culture medium, and seven chemical parameters of the soils were measured. Scedosporium species were detected in 48 of the samples, with the highest density in soils from wastewater treatment plants and landfills, followed by those from roadsides and polluted riverbanks, thus confirming the impact of human activities on their ecology. Scedosporium apiospermum was the most common species, followed by S. boydii and S. aurantiacum. Analysis of the chemical parameters of the soils revealed the presence of Scedosporium species was mainly associated with a moderate electrical conductivity, a pH range of 7.0 to 7.6, a nutrient-rich content and a moderate phosphorus amount. Thereby, these results demonstrated the relatively high occurrence of Scedosporium in Morocco and highlighted the impact of phosphorus content on their ecology.