ITS Genotypes Research Articles

Dermatophytosis Caused by Trichophyton indotineae (Trichophyton mentagrophytes ITS Genotype VIII) in Sri Lanka.

Over the past few years, a recalcitrant dermatophytosis has been observed on the Indian subcontinent, including Sri Lanka, which has caused a major public health issue in the region. An emerging species, Trichophyton indotineae, first described as Trichophyton mentagrophytes ITS genotype VIII, is thought to be responsible for this fast-spreading, mostly terbinafine-resistant dermatophytosis. Recalcitrant dermatophytosis is a challenge to dermatologists, and knowing the causative species and antifungal sensitivity in the earlier stage of management would be invaluable. We report a case series of patients with dermatophytosis caused by T. indotineae in Sri Lanka. This is the first detection of this highly terbinafine-resistant strain in Sri Lanka, and existence of this species should be taken seriously by dermatologists and healthcare policymakers for better management of tinea infections and antifungal stewardship in the country.

Dermatophytes and skin dermatophytoses in Southeast Asia-First epidemiological survey from Cambodia.

Dermatomycoses count to the most frequent dermatoses in Cambodia. The aim of this survey was to investigate the occurrence of dermatophytes in this Southeast Asian country. From June 2017 to July 2018, skin scrapings were taken from 67 patients with superficial dermatophytosis for mycological diagnostics. Identification of dermatophytes was confirmed by sequencing of the 'internal transcribed spacer'-(ITS) region of the rDNA, and the gene of the Translation Elongation Factor (TEF)-1α. Patients were suffering from tinea corporis and tinea inguinalis/cruris 42/67 (63%), tinea capitis/faciei 14/67 (21%), tinea corporis/capitis/faciei 6/67 (9%), tinea manuum/pedis 2/67 (3%), tinea pedis 2/67 (3%) and tinea manuum 1/67 (1%). Both, by culture and/or PCR, a dermatophyte was detected in 52 (78%) out of 67 samples. Culture positive were 42 (81%) of 52, PCR positive were 50 (96%). The following dermatophytes were found: Trichophyton (T.) rubrum, 36/52 strains (69%, 29 by culture), T. mentagrophytes/T. interdigitale (TM/TI) 9/52 (17%, six by culture) and Microsporum (M.) canis 5/52 strains (10%, by culture). One strain of Nannizzia (N.) incurvata 1/52 (2%) and N. nana 1/52 (2%) was isolated. Based on sequencing, we demonstrated that two T. mentagrophytes strains out of the nine TM/TI represented the new ITS genotype XXV Cambodia. We found one T. mentagrophytes strain genotype VIII (now, reclassified as T. indotineae). This isolate was terbinafine resistant, and it exhibited the amino acid substitution Phe397Leu in the squalene epoxidase. Three strains of T. interdigitale genotype II* were isolated. This is the first survey on epidemiology of dermatophytes in Cambodia. Currently, T. rubrum represents the most frequent species in Cambodia. One Indian strain genotype VIII T. mentagrophytes was found. A highlight was the first description of the new T. mentagrophytes genotype XXV Cambodia.

Molecular characterization of common zoonotic protozoan parasites and bacteria causing diarrhea in dairy calves in Ningxia Hui Autonomous Region, China.

Diarrhea caused by zoonotic pathogens is one of the most common diseases in dairy calves, threatening the health of young animals. Humans are also at risk, in particular children. To explore the pathogens causing diarrhea in dairy calves, the present study applied PCR-based sequencing tools to investigate the occurrence and molecular characteristics of three parasites (Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi) and three bacterial pathogens (Escherichia coli, Clostridium perfringens, and Salmonella spp.) in 343 fecal samples of diarrheic dairy calves from five farms in Lingwu County, Ningxia Hui Autonomous Region, China. The total positive rate of these pathogens in diarrheic dairy calves was 91.0% (312/343; 95% CI, 87.9-94.0), with C. perfringens (61.5%, 211/343; 95% CI, 56.3-66.7) being the dominant one. Co-infection with two to five pathogens was found in 67.3% (231/343; 95% CI, 62.4-72.3) of investigated samples. There were significant differences (p<0.05) in the positive rates of Cryptosporidium spp. and diarrheagenic E. coli among farms, age groups, and seasons. Two Cryptosporidium species (C. parvum and C. bovis) and five gp60 subtypes of C. parvum (IIdA15G1, IIdA20G1, IIdA19G1, IIdA14G1, and a novel IIdA13G1) were identified. Two assemblages (assemblage E and zoonotic assemblage A) of G. duodenalis and six ITS genotypes of E. bieneusi (J, Henan-IV, EbpC, I, EbpA, and ESH-01) were observed. Four virulence genes (eaeA, stx1, stx2, and st) of diarrheagenic E. coli and one toxin type (type A) of C. perfringens were detected. Our study enriches our knowledge on the characteristics and zoonotic potential of diarrhea-related pathogens in dairy calves.

Carbon amendments in soil microcosms induce uneven response on H2 oxidation activity and microbial community composition.

High-affinity H2-oxidizing bacteria (HA-HOB) thriving in soil are responsible for the most important sink of atmospheric H2. Their activity increases with soil organic carbon content, but the incidence of different carbohydrate fractions on the process has received little attention. Here we tested the hypothesis that carbon amendments impact HA-HOB activity and diversity differentially depending on their recalcitrance and their concentration. Carbon sources (sucrose, starch, cellulose) and application doses (0, 0.1, 1, 3, 5% Ceq soildw-1) were manipulated in soil microcosms. Only 0.1% Ceq soildw-1 cellulose treatment stimulated the HA-HOB activity. Sucrose amendments induced the most significant changes, with an abatement of 50% activity at 1% Ceq soildw-1. This was accompanied with a loss of bacterial and fungal alpha diversity and a reduction of high-affinity group 1h/5 [NiFe]-hydrogenase gene (hhyL) abundance. A quantitative classification framework was elaborated to assign carbon preference traits to 16S rRNA gene, ITS and hhyL genotypes. The response was uneven at the taxonomic level, making carbon preference a difficult trait to predict. Overall, the results suggest that HA-HOB activity is more susceptible to be stimulated by low doses of recalcitrant carbon, while labile carbon-rich environment is an unfavorable niche for HA-HOB, inducing catabolic repression of hydrogenase.

The Domestic Isolation of Terbinafine- and Itraconazole-Resistant Trichophyton indotineae in Chinese Mainland.

Trichophyton indotineae, a new species of dermatophytes, has become a significant concern in treating dermatophytosis due to the high level of terbinafine resistance reported in India and even worldwide. This study aimed to report the terbinafine- and itraconazole-resistant T. indotineae in Chinese mainland, by identifying the phylogenetic classification of the isolate strain, and detecting the drug resistance, gene mutation and expression. The skin scales of the patient were cultured on SDA and the isolate was authenticated by DNA sequencing and MALDI-TOF MS. Antifungal susceptibility testing was performed following the M38-A2 CLSI protocol to examine the MICs values of terbinafine, itraconazole, fluconazole, etc. The strain was screened for mutations in the squalene epoxidase (SQLE) gene by Sanger sequencing and detected the expression of CYP51A and CYP51B by qRT-PCR. A multi-resistant ITS genotype VIII sibling of the T. mentagrophytes complex (T. indotineae) was isolated in Chinese mainland. The strain harbored high terbinafine MIC of > 32μg/mL and itraconazole MIC of 1.0μg/mL, which was identified a mutation in the squalene epoxidase gene with amino acid substitution (Phe397Leu, mutation 1191C > A). In addition, overexpression of CYP51A and CYP51B was observed. With multiple relapses, the patient finally achieved clinical cure by itraconazole pulse therapy and topical clotrimazole cream for 5weeks. The first domestic strain of terbinafine- and itraconazole-resistant T. indotineae from a patient in Chinese mainland was isolated. Itraconazole pulse therapy can be an effective method for the treatment of T. indotineae.

The epidemic of the multiresistant dermatophyte Trichophyton indotineae has reached China.

Due to its high degree of natural resistance to terbinafine in vitro and its tendency to spread globally from the Indian subcontinent, the emerging dermatophyte Trichophyton indotineae has become a major concern in dermatology. Herein, we present the first report of T. indotineae from mainland China. The transmission of the fungus to Guizhou Province in central China and eventual host susceptibilities were investigated. We studied 31 strains of the T. mentagrophytes complex from outpatient clinics of our hospital collected during the past 5 years. The set comprised four ITS genotypes, two of which were T. mentagrophytes genotype VIII, now known as Trichophyton indotineae; the earliest isolation in the Guiyang area appeared to date back to 2018. The isolate was derived from an Indian patient, while local Chinese patients had no dermatophytosis caused by this genotype. Reports from around the world indicated that almost all of the globally reported T.indotineae cases originated from the Indian subcontinent and surrounding countries without transmission among native populations, suggesting deviating local conditions or racial differences in immunity against this fungus.

High genotype diversity and zoonotic potential of Enterocytozoon bieneusiinyaks (Bos grunniens) fromGanzi Tibetan Autonomous Prefecture, Sichuan Province.

Enterocytozoon bieneusi is a common pathogen in humans and various animals, threatening the breeding industry and public health. However, there is limited information on the molecular characteristics of E. bieneusi in yaks, an economically important animal mainly domesticated in the Qinghai Tibet Plateau in China. In the present study, nested PCR targeting the ITS gene region was applied to investigate the positive rates and genetic diversity of E. bieneusi in 223 faecal samples of yaks from three locations in Ganzi Tibetan Autonomous Prefecture, Sichuan Province. The total positive rate of E. bieneusi was 23.8% (53/223). Significant differences in positive rates were identified among yaks from three locations (χ2=8.535, p=0.014) and four age groups (χ2=17.259, p=0.001), with the highest positive rates in yaks from Yajiang and aged < 6 months, respectively. Sequence analysis identified seven known (EbpC, LW1, LQ10, PigEBITS5, ESH-01, J and BEB4) and five novel (Ganzi1-5) ITS genotypes. Phylogenetic analysis showed eight genotypes (EbpC, LW1, LQ10, PigEBITS5, ESH-01, Ganzi1, Ganzi2 and Ganzi4) in group 1 and three genotypes (J, BEB4 and Ganzi3) in group 2, indicating high genotype diversity and zoonotic potential of E. bieneusi in yaks from Ganzi. Considering the increasing zoonotic genotypes in yaks in the present study compared with previous findings, interventions should be developed to reduce the potential transmission of E. bieneusi between humans and animals.

Time and cost-efficient identification of Trichophyton indotineae.

During the past 5 years, an outbreak of recalcitrant dermatophytosis due to a novel Trichophyton species generally resistant to terbinafine, T.indotineae, has spread out from South Asia to many countries around the World. These isolates cannot be reliably differentiated from other Trichophyton spp. on the basis of morphological traits and the current laboratory diagnostics relies on sequencing of ribosomal DNA ITS region. In this study, we aimed to introduce two inexpensive and rapid PCR-based assays for differentiation between T.indotineae and other dermatophytes. The first introduced assay is based on PCR-restriction fragment length polymorphism (PCR-RFLP) analysis, involving the amplification of TOP2 sequences and the digestion of PCR products by Cfr13I restriction enzyme. The second assay is proposed as conventional endpoint species-specific PCR amplification of the C120-287 intergenic locus. To validate the assays, a total of 191 Trichophyton spp. and 2 Microsporum canis strains with known ITS region sequences were used. From the T.mentagrophytes / T.interdigitale species complex (TMTISC), strains with 18 different ITS genotypes were tested. The sample of TMTISC isolates included 41 T.indotineae strains. TOP2 PCR-RFLP and T.indotineae-specific PCR were positive with testing on DNA of all 41 T.indotineae isolates and two strains of T.mentagrophytes belonging to ITS Types XIII and XVI, but negative with other species and other TMTISC ITS genotypes (n=152). Therefore, the specificity of both new assays was 99%. The two developed diagnostic assays provide accurate and cost-effective means of identifying cultured T.indotineae isolates.

Tragopogon dubius: Multiple introductions to North America and the formation of the New World tetraploids

AbstractTragopogon (Asteraceae) is an evolutionary model for the study of whole‐genome duplication, with two recently and repeatedly formed allopolyploids, T. mirus and T. miscellus, and many additional polyploid species. Tragopogon mirus and T. miscellus formed in western North America following the introduction of three diploids from Europe: T. dubius, T. porrifolius, and T. pratensis. Of these diploids, T. dubius is a shared parent of both tetraploids and is broadly defined and widely distributed in Eurasia. Because human‐mediated intercontinental introductions may lead to hybridization with local species, and associated polyploidization, the introduction history of T. dubius from Europe to North America provides further opportunity to investigate both the extent and consequences of plant introductions. Using ITS sequence data, we show that the morphologically diverse, broadly defined T. dubius comprises a complex of at least 10 different ITS types in its native range, six of which have been introduced from Europe into North America. Significantly, although the two allotetraploid species have each formed multiple times on geographical scales from local to regional, recurrent formation is the result of repeated hybridization involving only one of these ITS subtypes. These results reinforce earlier data suggesting that not all diploid genotypes can form allopolyploids. Several entities traditionally considered part of T. dubius s.l. are now recognized as distinct species (e.g., T. lainzii), and it is likely that other distinct ITS genotypes identified here may also mark unique lineages that ultimately merit recognition as separate species. However, more intensive phylogenetic analyses involving many more samples from across the geographic range of T. dubius are required, as are detailed assessments of taxonomy, morphology, and cytology.

"Indian" strains of Trichophyton mentagrophytes with reduced itraconazole susceptibility in Germany.

We use published reports and three of our own tinea cases as an opportunity to report on "Indian" strains of Trichophyton (T.) mentagrophytes with ITS genotype VIII and reduced susceptibility to itraconazole due to the mutation c.1342G>A in the SQLE gene in Germany. In vitro measurements of resistance revealed normal susceptibility to terbinafine, but markedly reduced susceptibility to itraconazole - although no valid breakpoints are currently defined and minimum inhibitory concentrations (MICs) depend on the methods used. Problems related to the determination and interpretation of MICs are outlined. Our cases show that azole-resistant "Indian" strains of T. mentagrophytes with ITS genotype VIII occurred in Germany as early as 2011, which is earlier than was previously assumed. This variant of the pathogen cannot be phenotypically distinguished from customary strains of T. mentagrophytes; its identification is based on genetics. The taxonomic classification is still under debate. This variant is anthropophilic and causes only mildly inflammatory tinea lesions with many fungal elements. Its further dissemination must therefore be expected. Prerequisites for rapid and valid antimycotic testing against dermatophytes need to be developed.

Molecular-Based Detection of Enterocytozoon bieneusi in Farmed Masked Palm Civets (Paguma larvata) in Hainan, China: A High-Prevalence, Specificity, and Zoonotic Potential of ITS Genotypes.

Enterocytozoon bieneusi is a microsporidian and zoonotic species. This study investigated the prevalence and distribution of E. bieneusi genotypes in farmed masked palm civets using nested PCR, as well as assessed their zoonotic potential by phylogenetic analysis of the ITS region of the rRNA region. Here, we collected 251 fecal specimens from farmed masked palm civets (Paguma larvata) from the Hainan Island, China. In total, 128 of 251 samples were positive for E. bieneusi, with an average infection rate of 51.0%. Seventeen genotypes were identified including 12 known genotypes—HNR-VI (n = 56), SHR1 (n = 45), SHW7 (n = 6), KIN-1 (n = 3), D (n = 3), New1 (n = 3), EbpC (n = 2), CHC5 (n = 1), CHG19 (n = 1), CHN4 (n = 1), EbpA (n = 1), and Henan-III (n = 1)—and five novel genotypes (HNPL-I to HNPL-II; one each). Phylogenetic analysis categorized these genotypes into two groups. Thirteen of them were members of the zoonotic group 1, and the remaining four genotypes were in group 12. This study has shown that the infection rates of E. bieneusi in masked palm civets from Hainan were relatively high and provide baseline data to control and prevent microsporidiosis in farm-related communities. Therefore, infections in masked palm civets with zoonotic genotypes D, EbpC, CHN4, EbpA, KIN-1, and Henan-III should be considered potential threats to public health.

Genotyping of Ascaris spp. infecting humans and pigs in Italy, Slovakia and Colombia.

The systematics and taxonomy of Ascaris lumbricoides and Ascaris suum, two of the world's most widespread nematodes, still represent a highly debated scientific issue. Two different transmission scenarios have been described according to endemicity: separated host-specific transmission cycles in endemic regions, and a single pool of infection shared by humans and pigs in non-endemic regions. The swine roundworm A. suum is now recognized as an important cause of human ascariasis also in endemic areas such as China, where cross-infections and hybridization have also been reported, as well as in non-endemic regions like Italy. This study aimed to investigate the molecular epidemiology of human and pig ascariasis in three countries representing different epidemiological scenarios: Italy as a non-endemic country, Colombia as an endemic country, and Slovakia as a non-endemic country, but with a poor socio-economic context linked to some focal populations of Roma settlements. A total of 237 nematodes were analysed: 46 from Colombia (13 from humans, 33 from pigs), 114 from Slovakia (20 from humans, 94 from pigs) and 77 from Italy (17 from humans and 60 from pigs). Genotyping by PCR-RFLP of nuclear (ITS) and sequencing of mitochondrial (cox1) target regions were performed. ITS genotypes were used to estimate the Hardy-Weinberg (HW) equilibrium according to hosts and country of origin. The partial cox1 sequences were used to analyse genetic polymorphisms according to hosts and country of origin, as well as to infer the network of haplotypes, their evolutionary relationships and geographical distribution. 110 quality cox1 sequences were obtained. Haplotype network revealed three main groups corresponding to clade A, B and C. Clade C included most of the human cases from Italy, while those from Slovakia and Colombia were grouped in clade B. Ascaris from Italian and Colombian pigs showed HW equilibrium at the ITS marker, while disequilibrium was found in A. lumbricoides from Slovak pigs, which suggest a high unexpected amount of roundworms of human origin circulating also in pigs. This study updates and extends the current understanding of Ascaris species and genotypes circulating in different epidemiological scenarios, with particular attention to the inclusion of human-derived Ascaris in the phylogenetic cluster C. Despite the evidence of HW equilibrium in the ITS in pig-derived Italian samples, the amount of genetic variation seems to support the existence of two closely related species.

Risk Evaluation of Pathogenic Intestinal Protozoa Infection Among Laboratory Macaques, Animal Facility Workers, and Nearby Villagers From One Health Perspective.

Background: Previous epidemiological studies have confirmed non-human primates (NHPs) as reservoirs for Cryptosporidium spp. , Giardia intestinalis, and Enterocytozoon bieneusi. It highlights the possibility of interspecies transmission between humans and macaques in laboratory animal facilities. This study aimed to investigate the prevalence of pathogenic intestinal protozoan infections in macaques and humans and to determine the risk of cross-species transmission from One Health view.Materials and Methods: A total of 360 fecal samples, including 310 from the four Macaca mulatta groups, 25 from the facility workers in a laboratory animal facility, and 25 from the villagers nearby in Yongfu country, southeast China, were collected. Nested PCR assays were done for detecting protozoan pathogens from all the specimens. Furthermore, potential risk factors (gender, age, and direct contact) on the occurrence of intestinal protozoa infection among different sub-groups were evaluated. A phylogenetic and haplotype network analysis was conducted to examine the genetic structure and shared patterns of E. bieneusi and Cyclospora cayetanensis.Results: The pathogenic intestinal protozoa were detected in both human and macaque fecal samples. A total of 134 (37.2%) samples were tested positive, which included 113 (36.4%) macaques, 14 (56.0%) facility workers, and 7 (28.0%) villagers, respectively. There was no significant difference in four intestinal protozoa infections between facility workers and villagers (χ2 = 2.4, P > 0.05). However, the positive rate of pathogenic intestinal protozoa in the facility workers, who had direct contact with macaques, was significantly higher [odds ratio (OR) = 0.31, 95% confidence interval (CI): 0.09–1.00, P < 0.05).Thirty-three ITS genotypes of E. bieneusi were identified, including five known genotypes (PigEBITS7, Peru8, Henan V, D, and CM1) and six novel genotypes (MEB1–6). Seven haplotypes were identified in the network analysis from C. cayetanensis-positive samples. Meanwhile, a phylogenetic and haplotype analysis confirmed the presence of zoonotic subtypes in NHPs and humans.Conclusion: The data collected from this study confirmed a high prevalence of intestinal protozoan infection in humans and macaques. These results warrant workers of such facilities and residents to limit contact with infected animals in order to minimize related health risks. The need for comprehensive strategies to mitigate the risk of zoonotic transmission, especially from a One Health perspective, is recommended.

The unprecedented epidemic-like scenario of dermatophytosis in India: II. Diagnostic methods and taxonomical aspects.

Trichophyton (T.) mentagrophytes now accounts for an overwhelming majority of clinical cases in India, a new "Indian genotype" (T. mentagrophytes ITS genotype VIII) having been isolated from skin samples obtained from cases across a wide geographical distribution in this country. The conventional diagnostic methods, like fungal culture, are, however, inadequate for diagnosing this agent. Thus, molecular methods of diagnosis are necessary for proper characterization of the causative agent. The shift in the predominant agent of dermatophytosis from T. rubrum to T. mentagrophytes, within a relatively short span of time, is without historic parallel. The apparent ease of transmission of a zoophilic fungus among human hosts can also be explained by means of mycological phenomena, like anthropization.

Genotyping and zoonotic potential of Cryptosporidium and Enterocytozoon bieneusi in pigs transported across regions in China

Cryptosporidium spp. and Enterocytozoon bieneusi are common and important enteric parasites that can infect humans and animals, causing diarrhoea and systemic diseases. The objectives of the present study were to examine the prevalence and genetic variations of Cryptosporidium and E. bieneusi in pigs transferred from northeastern China to Ningbo city in Zhejiang Province. Cryptosporidium spp. was detected in 0.9% (2/216) of these samples and belonged to the zoonotic species Cryptosporidium parvum. A high E. bieneusi infection rate (25.0%, 54/216) was observed in this study, with 7 possible novel ITS genotypes (JLNB-1 to JLNB-7) and 10 known genotypes (EbpA, CM11, H, CM6, pigEBITS1, EbpC, CS-4, pigEBITS5, CHS5, and Henan-Ⅳ) identified, and zoonotic EbpA was the dominant genotype. Genotypes H and pigEBITS1 were reported for the first time in pigs in China. Phylogenetic analysis indicated that all the genotypes found in these samples belonged to zoonotic group 1. These findings indicated the potential threat of Cryptosporidium and E. bieneusi to humans or the environment during cross-regional transportation. An effective management control system should be built to avoid parasitic transmission as well as other animal diseases while travelling across different regions. In further studies, attention should be given to the transmission routes and the role of pigs as a potential source of human Cryptosporidium and E. bieneusi infections in China.

Indian Trichophyton mentagrophytes squalene epoxidase erg1 double mutants show high proportion of combined fluconazole and terbinafine resistance.

The Indian ITS genotype VIII Trichophyton mentagrophytes population shows a high amount of different erg1 (ergosterol) mutants encoding for squalene epoxidase, which catalyses the first step of ergosterol biosynthesis. Illumination of the implication of point mutations at position Ala448Thr in single and double erg1 Tmentagrophytes mutants because mutants of this type were abundantly found within the Indian fungal population. Growth in fluconazole or terbinafine containing medium was analysed using a microplate-laser-nephelometry (MLN)-based growth assay. Ala 448 Thr erg1 single mutants were terbinafine sensitive, but about 50% of isolates showed an increased fluconazole resistance, whereas 95% of the double mutants (Phe 397 Leu, Ala 448Thr) demonstrated combined terbinafine and increased fluconazole resistance. The new Indian Tmentagrophytes populations show several point mutations in erg1. Point mutations at position 397 were previously described and cause terbinafine resistance. A large part of the double mutants exhibit resistance to terbinafine and fluconazole, demonstrating a selective advantage of the combination of both mutations.

Multilocus Sequence Typing of Enterocytozoon bieneusi Isolates From Various Mammal and Bird Species and Assessment of Population Structure and Substructure.

Enterocytozoon bieneusi is one of the most common intestinal pathogens in humans and animals. E. bieneusi has been confirmed to be complex microsporidian species. Approximately 500 ITS genotypes of E. bieneusi have been defined. With the establishment and application of multilocus sequencing typing and population genetic tools in E. bieneusi, the studies on these aspects have been carried out worldwide, but little information is available. To understand genetic variation of mini-/micro-satellites and the population structure and substructure of E. bieneusi in northeastern China, 305 E. bieneusi DNA specimens composed of 28 ITS genotypes were from 13 mammal species and five bird species in the investigated areas. They were characterized by nested-PCR amplification and sequencing at four mini-/micro-satellite loci (MS1, MS3, MS4, and MS7). At the MS1, MS3, MS4, and MS7 loci, 153 (50.16%), 131 (42.95%), 133 (43.61%), and 128 (41.97%) DNA specimens were amplified and sequenced successfully with 44, 17, 26, and 24 genotypes being identified, respectively. Multilocus genotypes (MLGs) showed a higher genetic diversity than ITS genotypes. 48 MLGs were produced out of 90 ITS-positive DNA specimens based on concatenated sequences of all the five genetic loci including ITS. Linkage disequilibrium (LD) and limited genetic recombination were observed by measuring LD using both multilocus sequences and allelic profile data, indicating an overall clonal population structure of E. bieneusi in the investigated areas. These data will aid in the longitudinal tracking of the attribution of source of infection/contamination and in elucidating transmission dynamics, and will provide valuable information for making efficient control strategies to intervene with and prevent occurrence of microsporidiosis caused by E. bieneusi among animals and transmission of E. bieneusi from animals to humans in the investigated areas. Phylogenetic and network analyses identified three different subgroups, revealing the presence of host-shaped segregation and the absence of geographical segregation in E. bieneusi population. Meanwhile, the MLGs from zoonotic ITS genotypes were observed to be basically separated from the MLGs from host-adapted ones. Assessment of substructure will have a reference effect on understanding of zoonotic or interspecies transmission of E. bieneusi and evolution direction from zoonotic genotypes to host-adapted genotypes.

Enterocytozoon bieneusi in donkeys from Xinjiang, China: prevalence, molecular characterization and the assessment of zoonotic risk

BackgroundEnterocytozoon bieneusi, a zoonotic pathogen, has the potential to infect both immunocompromised and immunocompetent humans. It is found in large number of animals; however, not much is known regarding its prevalence in equine animals, particularly donkeys. This is the first molecular epidemiological evaluation of E. bieneusi in 178 free-ranging donkeys from five countrysides; and 502 farmed donkeys from 18 farms in 12 cities of Xinjiang, China by Nested PCR.ResultsE. bieneusi was detected in 2.5% (17/680) donkeys, with 2.6% (13/502) in farmed and 2.2% (4/178) in free-ranging ones. Sequence analysis identified eight ITS genotypes, all belonging to zoonotic Groups 1 or 2, including six known genotypes: horse1 (n = 5), D (n = 3), NCD-2 (n = 3), BEB6 (n = 2), BEB4 (n = 1), and NIAI (n = 1); and two new genotypes: XJD1 (n = 1) and XJD2 (n = 1).ConclusionsThis is the first report confirming the presence of E. bieneusi in donkeys in Xinjiang, China, and indicates the possibility of zoonotic transmission of this pathogenic parasite.

Involvement of Little Discussed Anatomical Locations in Superficial Dermatophytosis Sundry Observations and Musings.

A rather quick change in epidemiological shift from Trichophyton rubrum to Trichophyton mentagrophytes (ITS genotype VIII), several changes in clinical presentation of superficial dermatophytoses and the the response/ the lack of it, makes this current epidemic very interesting and novel. We present here some of the relatively less discussed variants that have been described under the broad terms of tinea corporis and tinea faceie. Many of them are found to occur uncharacteristically frequently and some in areas that were never described. We describe some representative cases of female genital dermatophytoses, periumblical and periocular dermatophytosis and discuss some unusual variants like those showing sole involvement of the ear (tinea auricularis), lip (tinea labialis) and superficial dermatophytosis of the scalp skin.

Molecular detection of Cryptosporidium and Enterocytozoon bieneusi in dairy calves and sika deer in four provinces in Northern China.

The protistan pathogens Cryptosporidium and Enterocytozoon bieneusi can cause significant intestinal diseases in animals and humans. However, limited information is available regarding prevalence and molecular characterization of Cryptosporidium and E. bieneusi in ruminants in Northern China. In this study, the overall prevalence of Cryptosporidium and E. bieneusi was 19.3% (62/321) and 28.97% (93/321) in dairy calves and 1.10% (9/818) and 13.57% (111/818) in sika deer (Cervus nippon) in four provinces in Northern China, respectively. The prevalence of Cryptosporidium and E. bieneusi in different factor groups was various. Five Cryptosporidium species/genotypes were identified, of which C. parvum, C. ryanae, C. bovis, and C. andersoni were only found in dairy calves, and only Cryptosporidium deer genotype was found in sika deer. Moreover, J, I, and BEB4 ITS genotypes of E. bieneusi were found in dairy calves, and six known genotypes (JLD-III, JLD-IX, JLD-VII, EbpC, BEB6, and I) and ten novel genotypes (namely LND-I and JLD-XV to JLD-XXIII) were found in sika deer in this study. Cryptosporidium parvum and E. bieneusi genotype J were identified as the predominant species/genotypes in dairy calves, whereas the predominance of Cryptosporidium spp. and E. bieneusi in sika deer was Cryptosporidium deer genotype and BEB6, respectively. The present study reported the prevalence and genotypes of Cryptosporidium and E. bieneusi in dairy calves and sika deer in four provinces in northern China. The present findings also suggest that investigated dairy calves and sika deer may play an important role in the transmission of E. bieneusi and Cryptosporidium to humans and other animals, and also in an effort to better understand the epidemiology of these enteric pathogens in China.