Changes of Dermatophytoses in Southwestern Greece: An 18-Year Survey

The isolation and distribution rate of dermatophytes as causative agents of superficial mycoses of skin, hair, and nails during an 18-year period (1991–2008) at a university hospital are presented. A comparative analysis of epidemiological differences within the first (1991–1999) and the second 9-year period (2000–2008) was performed. Skin scrapings, nail, and hair specimens were examined by a direct microscopic examination and culture. Identification of dermatophyte species was based on macroscopic and microscopic characteristics of colonies. During the complete period (18 years), 5,971 patients with suspected dermatophytosis were examined. Seven hundred and sixty-nine patients (12.8%) were found positive. Among them, 495 cases (64.3%) were of skin dermatophytoses, 91(11.8%) of hair, and 183 (23.7%) of nails. The most frequent etiological agents were Microsporum canis (54%), Trichophyton rubrum (38%), and T. mentagrophytes (6%). Epidermophyton floccosum, T. tonsurans, T. violaceum, and M. gypseum were responsible only for 16 cases (2%) of dermatophytoses. The prevalence of dermatophytoses seems to decrease significantly from 16.2% (1991–1999)–9.6% during the last 9-year period. The most frequent dermatophyte, M. canis, shows decreasing trends during the last period (from 58.5 to 45.7%), whereas T. rubrum shows an increasing isolation rate (from 35 to 43.6%), respectively. The most common form of dermatophytosis among children remains tinea capitis due to M. canis. The most frequent etiological agent of tinea unguium (81%) is T. rubrum.     

Phylogenetic analysis of Altai osmans of the genus <Emphasis Type="Italic">Oreoleuciscus</Emphasis> (Pisces,Cyprinidae, Leuciscinae), based on the analysis of the cytochrome oxidase 1 gene (<Emphasis Type="Italic">Co-1</Emphasis>) sequence

Molecular genetic analysis of Altai osmans of the genus Oreoleuciscus from two different parts of the range was carried out. In this study, based on the mitochondrial Co-1 gene sequence, a total of 25 fish specimens belonging to four genera were examined: (1) O. humilis, 2 specimens; O. potanini, 13 specimens; (2) Pseudaspius leptocephalus, 1 specimen; (3) Tribolodon brandtii, T. hakonensis, and T. sachalinensis from the GanBank database, 9 speciens; and (4) Leuciscus waleckii, 1 specimen (used as an outgroup). The p-distances were very low both within and between the species: (1) 0.20 ± 0.03%; (2) 0.40 ± 0.12%; and (1–2) 0.80 ± 0.04%. To visualize the relationships among all of the species examined, the neighbor joining (NJ), maximum parsimony (MP), Bayesian (BA), and maximum likelihood (ML) trees were constructed. The results obtained using these methods were very similar. It was demonstrated that species assignment of the individuals (barcoding) with the help Co-1 gene was effective, despite of very low divergence of the two osman taxa, which was comparable with typical intraspecific values in other animal groups. Taxonomic status of O. potanini and O. humilis requires further investigation with paying attention to low genetic distances between these species along with the lack of material from sympatric parts of the ranges.     

Polymorphism of SLC14A1 encoding human erythrocytic Kidd antigens in the Chinese population

The SLC14A1 gene, which encodes the important Kidd blood group antigens, has not been systematically?analyzed at the molecular level in Chinese individuals. In this study, SLC14A1 genetic polymorphism was examined in Chinese individuals with Jk(a+b-), Jk(a+b+), and Jk(a-b+) expression. The Kidd phenotype was determined for 146 specimens using monoclonal anti-Jk^a and -Jk^b antibodies. From these, 87 specimens were Jk(a-b+), 21 were Jk(a+b-), and 38 were Jk(a+b+). According to the Kidd phenotype results, 20 specimens were randomly selected from each group, i.e., Jk(a-b+), Jk(a+b-), and Jk(a+b+), for the molecular analyses of exons 3 to 11 of the SLC14A1 gene. Novel alleles were detected in the SLC14A1 gene, including IVS3-106A, IVS3-99A, exon3 130G, IVS4-299G, IVS4-293G, IVS4+211C, IVS4 +230C, exon6 499A, exon6 588A, IVS7-68T, IVS9+244G, and IVS10-153T, indicating that the locus harbored significant polymorphism. We also showed that IVS4-299, IVS7-68, and IVS10-153 were novel SNPs absolutely associated with exon 8 nt. 838. The minor allele frequencies were all greater than 10% and all SNPs in the Chinese population showed Vel antigen expression on RBC membranes. We identified 12 SNPs in the SLC14A1 gene in the Chinese population, IVS3-106A, IVS3-99A, exon3 130G, IVS4-299G, IVS4-293G, IVS4+211C, IVS4 +230C, exon6 499A, exon6 588A, IVS7-68T, IVS9+244G, and IVS10-153T. Our results also indicated that three novel SNPs produced Jk^a and Jk^b antigens in Chinese individuals.     

A ten years survey of Romanian soil screening for keratinophilic fungi (1958–1967)

Summary Out of 750 soil samples collected from various regions of Romania, during 1958–1967, a positive percentage of 86 was positive for keratinophilic fungi. Among the 13 isolated species, the most prevalent wereM. gypseum (54.6 %),C. keratinophilum (40.1 %) andK. (T.)ajelloi (27 %). BesideT. terrestre, found in 8.9 % of the examined soil samples, the percentage of the remaining species is low, not exceeding 4 %. In order of frequency:C. (T).evolceanui (3.2 %),M. cookei (2.2 %),T. mentagrophytes (2 %),T. quinckeanum (0.9 %),C. (T.)indicum (0.6 %). There have been also isolated, but only once;M. canis, A. multifidum, T. georgiae andT. vanbreuseghemii.The studies carried out on the immunobiological properties of the Romanian soil isolates (including intracutaneous tests, serological reactions and therapeutic efficiency), have showed a close relationship between these fungi and the common pathogenic dermatophytes. The pathogenic power of the keratinophilic fungi is briefly discussed.     

Keratinophilic fungi isolated from soil of the Abruzzo National Park,Italy

One hundred sixty one soil samples from the Parco Nazionale d'Abruzzo were analyzed. The keratinophilic fungi present in the soil samples were isolated and classified with the hair-bait method. The most common was T. ajelloi (28.6% of the total samples) followed by Chrysosporium keratinophilum (14.3%), C. sp. (11.2%), C. asperatum and Trichophyton sp. (6.8%), T. terrestre (5.6%), C. pannorum and C. state of Ctenomyces serratus (3.7%), C. tropicum (3.1%), M. cookei (2.5%) and the C. state of Arthroderma tuberculatum (1.9%). Other species, not belonging to the dermatophytes, were also isolated. The absence of dermatophytes was correlated with low human and animal population density.Altitude and vegetation did not seem to influence their distribution.     

Biodegradation of Crude Oil by Constructed Bacterial Consortia and the Constituent Single Bacteria Isolated From Malaysia

Three bacterial isolates identified as Pseudomonas aeruginosa (UKMP-8T), Rhodococcus sp. M15-2 (UKMP-5T), and Rhodococcus sp. ZH8 (UKMP-7T) based on biochemical, physiological, and morphological characteristics and on 16S rDNA sequences were isolated from groundwater of a crude oil refinery plant. From these three isolates, four bacterial consortia were designed by mixing the single bacterial cultures in the following ratios: (P. aeruginosa:Rhodococcus sp. M15-2, 1:1), (P. aeruginosa:Rhodococcus sp. ZH8, 1:1), (Rhodococcus sp. M15-2: Rhodococcus sp. ZH8, 1:1), and (P. aeruginosa: Rhodococcus sp. ZH8:Rhodococcus sp. M15-2, 1:1:1), respectively. Bacterial isolates and consortia showed differing preferences for nitrogen source (0.01% ammonium chloride, 0.10% yeast extract, or 0.50% peptone) to reach optimum growth. When fortified with the preferred nitrogen sources and grown in minimal salt medium, within 7 days all three single isolates and the four bacterial consortia biodegraded 97.6-99.9% of Tapis Massa oil without any significant differences.     

广西植物标本馆馆藏模式标本现状与分析

模式标本是植物分类和命名的依据和凭证,也是植物资源保护与持续利用的重要基本资料。为提高模式标本的管理水平,促进其充分利用和互惠共享,该文对广西植物标本馆(IBK)馆藏模式标本的种类组成、采集地、采集年代及采集人等方面进行了统计和分析。结果表明:(1)本馆完成信息数字化的模式标本共计4 645号,5 736份,隶属于170科619属2 128种(含种下等级),馆藏模式标本收集的优势科有樟科、苦苣苔科、茜草科、山茶科、兰科、荨麻科、百合科、猕猴桃科、大戟科、柿科和杜鹃花科等,优势属有秋海棠属、蜘蛛抱蛋属、马兜铃属、紫金牛属和冬青属等。(2)模式标本分别采自中国的20个省(区、市)以及其他2个国家,模式标本采集较多的省区有广西(48.19%)、海南(24.14%)和广东(11.63%)。(3)历年模式标本采集高峰期在20世纪30年代和50年代,模式标本命名高峰期在20世纪80年代,与我国植物分类学的兴起,以及开展《中国植物志》编研等有关,模式标本的橱柜时间(标本采集到命名的时间间隔)平均为21年,橱柜时间最长为82年。该文还对模式标本的馆藏现状及数字化工作进行了讨论和展望。     

Cold activity and tolerance of the entomopathogenic fungus Tolypocladium spp. to UV-B irradiation and heat

Studies on the stress resistance of insect-pathogenic fungi are very important to better understand the survival of these organisms in the environment. In this study, we examined the cold activity (8 ± 1 °C for 7 days), UV-B tolerance (Quaite-weighted UV-B irradiance at 847.90 mW m⁻² for 1, 2, 3, and 4 h), and wet-heat tolerance (45 °C for 1, 2, 3, and 4 h) of two isolates of Tolypocladiumcylindrosporum (ARSEF 3392 and 5558), one isolate of Tolypocladium geodes (ARSEF 3275), and two isolates of Tolypocladium inflatum (ARSEF 4772 and 4877) based on their germination, compared with Metarhizium robertsii (ARSEF 2575). After 3 h of UV-B exposure, T. cylindrosporum germinated at a greater rate than the other Tolypocladium species and had similar viability to that of the M. robertsii. Most Tolypocladium isolates, however, were less UV-B tolerant than M. robertsii. The T.cylindrosporum isolates were also the most thermotolerant, with similar tolerance to the M. robertsii. The isolates of T. inflatum and T. geodes, which had similar heat tolerance, were the least heat tolerant compared with the isolates of T. cylindrosporum and M. robertsii. After 4 h of heat exposure, the germination of T. inflatum and T. geodes isolates was not significantly different. For cold activity, both T.cylindrosporum isolates germinated to ca. 100% in only 3 days. Approximately 50% of the two T. inflatum isolates germinated, and less than 5% of T. geodes germinated after 3 days. All fungal isolates, however, completely germinated by the seventh day, except M.robertsii. The isolates of T. cylindrosporum, therefore, were the most heat and UV-B tolerant, and had the highest cold activity compared to the other species. The tolerance of M. robertsii to UV-B radiation and heat was similar to that of T.cylindrosporum.     

Tinea Capitis in Southeastern China: A 16-Year Survey

This survey was a retrospective of a 16-year (1993–2008) study on the incidence, clinical features, and etiological agents of tinea capitis mainly representing the Southeastern China. The diagnosis was confirmed by direct microscopic examination. Eight hundred and sixty-six patients with tinea capitis, 381 males (44%) and 485 females (56%), were enrolled in this study. Patients were between 20 days and 84 years old with an average of 10.5 years and the peak incidence was in the age group of 6–10 (48.5%). Five hundred and sixty-two patients (64.9%) were ectothrix and 303 patients (35.0%) were endothrix with only one patient was favus. The incidence of tinea capitis from 1993 was gradually increasing and reaching to its peak in 2001. Positive cultures of dermatophytes were obtained in 715 patients: Microsporum canis (62.4%) was predominant, followed by Trichophyton violaceum (19.0%), Trichophyton tousurans (9.8%). M. canis was the major pathogen for ectothrix infection, while T. violaceum and T. tousurans contributed to the most endothrix form. M. canis, T. violaceum, and T. rubrum were the major pathogens for kerion.     

KRAS and BRAF mutation analysis can be reliably performed on aspirated cytological specimens of metastatic colorectal carcinoma

N. K. B. Pang, M. E. Nga, S. Y. Chin, T.‐M. Ismail, G. L. Lim, R. Soong and M. Salto‐Tellez
KRAS and BRAF mutation analysis can be reliably performed on aspirated cytological specimens of metastatic colorectal carcinoma Background: Sanger sequencing is one of several reliable methods in use to detect KRAS and BRAF mutations to facilitate clinical patient selection for anti‐epidermal growth factor receptor (EGFR) monoclonal antibody therapy in unresectable metastatic colorectal adenocarcinoma (CRC). Most analyses are made on pretreatment biopsy or resection specimens. There is a scarcity of published studies on the suitability of cytological samples for KRAS testing in this setting. Methods: DNA extraction was attempted on 11 search‐retrieved paired cases of histological resections or excisions of CRC and their corresponding cytological samples (representing metastases) and tested for KRAS mutations in exon 2 and 3, as well as BRAF exon 15 mutations by Sanger sequencing. Only KRAS wild‐type cases were subjected to BRAF analysis because this is the setting with true diagnostic value, as these mutations are mutually exclusive. Results: Of the 11 paired cases analysed, only eight histology cases showed satisfactory DNA quality for sequencing. Thus, only eight of the corresponding cytology cases were analysed. Seven of the eight cases tested showed the same KRAS genotype on both the aspirated cytology specimen of metastatic carcinoma and the primary tumour (histological specimen), from which we derive an overall concordance rate of 87.5%. The single discordant case was likely to be a true difference as it was demonstrated again on repeat testing of both samples. No BRAF mutations were detected on the four KRAS wild‐type cases. Conclusion: A range of cytological samples are suitable for KRAS and BRAF mutation testing, be it from previously stained preparations or cell blocks. These samples would be highly valuable in cases where cytological samples are the only material available for mutation testing.     

A molecular screening assay to identify Chlamydia trachomatis and distinguish new variants of C. trachomatis from wild-type

Chlamydia trachomatis is the most common sexually transmitted pathogen globally, causing serious health problems and representing a burden on public health. A new variant of C. trachomatis (nvCT) that carries mutations (C1514T, C1515T and G1523A) in the 23S rRNA gene has eluded detection in Aptima Combo 2 assays. This has led to false negatives in diagnostics tests and poses a challenge for C. trachomatis diagnostics on a global level. In this study, we developed a simple and cost-effective assay to identify C. trachomatis, with a potential application to screen for nvCT. We developed a screening assay based on high-resolution melting (HRM), targeting the 23S rRNA gene and cryptic plasmid. To evaluate the performance of the assay, 404 archived C. trachomatis DNA specimens and 570 extracted clinical specimens were analysed. Our HRM assay not only identified C. trachomatis in clinical specimens, but also correctly differentiated nvCT carrying C1514T, C1515T and G1523A mutations from the wild-type. We observed no cross-reactions with other clinically related agents, and the limit of detection was 11.26 (95% CI; 7.61–31.82) copies per reaction. Implementation of this screening assay could reduce detection times and costs for C. trachomatis diagnoses, and facilitate increased research on the presence and monitoring of nvCT.     

Optimization of formation and regeneration of protoplasts from biocontrol agents ofTrichoderma species

The optimal conditions necessary for a large yield and a high frequency of regeneration of protoplasts isolated from the biocontrol agentsTrichoderma koningii andT. harzianum were investigated. Protoplast yields were 1.2×10⁸/ml fromT. koningii and 6×10⁷/ml fromT. harzianum when 20-h mycelial culture was treated with a lytic enzyme solution containing Novozym 234 (15 mg/ml), sucrose (0.6 M) and citrate phosphate buffer (0.02 M), pH 5.6 at 31°C. When the protoplasts were grown in the regeneration medium containing yeast extract (1.5%), 1 I of Mandel's salts, pH 5.6, and glucose (0.8 M), a high frequency of regeneration of the protoplast was obseved: 66% forT. koningii and 45% forT. harzianum. Two patterns of regeneration were observed. First, the hyphae arose directly from the regenerated protoplast mother cell. Second, a chain of bud cells developed from the protoplast and subsequently generating hyphae generally protruded from the terminal bud cells.     

The musculus-type Y Chromosome of the laboratory mouse is of Asian origin

Mus musculus domesticus, M.m. bactrianus, M. m. musculus, M.m. castaneus, and M.m. molossinus wild mice were investigated for polymorphisms of the Y Chromosome (Chr) genes Zinc finger-Y (Zfy) and Sex-determining region-Y (Sry). Zfy divided the Y Chrs of these mice into domesticus- (domesticus) and musculus-types (musculus, castaneus, molossinus). M.m. bactrianus specimens had both Y Chrs, possibly owing to the introgression of a musculus-type Y into this population. Sry identified a subpopulation of musculus-type Y chromosomes. This subpopulation, designated the molossinus-type, was found in M.m. molossinus, a M. musculus subspecies specimen from northern China (Changchun), and laboratory mice. The cumulative data suggest that M.m. musculus of northern China and Korea are subpopulation distinct from M.m. musculus of Europe and central China and that this subpopulation invaded Japan, giving rise to M.m. molossinus. Furthermore, the data suggest that the musculus-type Y of the laboratory mouse originated from this subpopulation, corroborating early historical record reporting that Chinese and Japanese mice that were imported into Europe for the pet trade contributed to the genome of the laboratory mouse.     

贵州六盘水发现东阳江麝鼩

在贵州省六盘水市杨梅乡慕尼克村,利用陷阱法捕捉到3号麝鼩属(Crocidura)标本。本次采集标本的体形较小,头体长(49.0 ± 0.8)mm,尾长[(41.8 ± 4.2)mm]略短于头体长(尾长/头体长为85%)。背毛呈浅灰褐色,腹毛颜色浅于背毛,呈灰色。尾部双色,背侧黑褐色,腹侧淡于背侧。前足背部白色,后足则为淡灰色。尾近乎裸露,尾基约1/3着生稀疏白色长毛。颅全长(15.92 ± 0.55)mm,脑颅高(4.75 ± 0.18)mm。上门齿1枚,有一长而大的前尖和一小而矮的后尖。上单尖齿3枚,第1单尖齿最大,第2单尖齿略大于第3单尖齿,1枚第四前臼齿(P4),3枚臼齿。上述特征与东阳江麝鼩(C. dongyangjiangensis)模式标本的描述和鉴定特征基本一致,因此将3号采集标本鉴定为东阳江麝鼩。基于Cyt b基因进行分子系统发育分析,采集标本与麝鼩属物种中的东阳江麝鼩遗传距离最近,在0.004 ~ 0.027之间。系统发生树显示,3号标本与东阳江麝鼩构成一个单系进化分支,进一步证实本次采集的3号标本是东阳江麝鼩,为贵州省分布新记录种。     

A short review on the onychomycoses by dermatophytes in Romania

In 525 patients with onychomycoses by dermatophytes, collected in Bucharest (45% of the cases being females), the toe-nails were found to be affected in 65% of the cases, while the finger-nails only in 20%. In order of frequency, the following agents were isolated:T. rubrum (55%),T. mentagrophytes (interdigitale — 26%),T. violaceum (12%),E. floccosum (3%),T. schonleinii (1.3%). Only sporadically were found other species, viz.:M. canis (4),T. tonsurans (3),T. verruccosum (2),T. quinckeanum (1) andM. audouinii (1). Tinea pedis was the main manifestation associated with the respective cases.     

Genetic evidence for a complex of species within Rugopharynx australis (Mönnig, 1926) (Nematoda: Stronglyloidea) from macropodid marsupials

An electrophoretic analysis using 17 enzyme loci was carried out on specimens of the gastric nematode of macropodid marsupials, Rugopharynx australis (Mönnig, 1926), collected from Macropus eugenii (Desmarest), M. fuliginosus (Desmarest), M. giganteus Shaw, M. robustus Gould, M. rufogriseus (Desmarest), M. rufus (Desmarest), Thylogale billardierii (Desmarest) and Wallabia bicolor (Desmarest) from south-eastern Australia. The extent of fixed genetic differences between nematodes from different host species ranged from 0–53%. The two distinct morphological forms of the parasite found in M. rufogriseus differed at 50% of loci. Specimens present in M. fuliginosus and M. giganteus were indistinguishable genetically, as were nematodes from M. rufus and M. robustus. Of the two morphologically distinct congeners included in the analysis as controls, Rugopharynx epsilon (Johnston & Mawson, 1939) was genetically distinct (46–69% fixed genetic differences) from all specimens of the R. australis complex while R. rufogrisea Magzoub, 1964 was closely related to one of the two species occuring in M. rufogriseus. It was concluded that R. australis is a species complex, with a genetically distinct species present in M. eugenii, M. fuliginosus/M. giganteus, M. robustus/M. rufus, W. bicolor and T. billardierii, and two species in M. rufogriseus.     

Reinstatement of Myelophycus caespitosus (Scytosiphonaceae,Phaeophyceae) from Japan

Reinstatement of Myelophycus caespitosus Kjellman (Ectocarpales s.l., Phaeophyceae) described from western Japan is proposed based on the comparisons of DNA sequences of Japanese Myelophycus specimens, the type specimen of Chordaria simplex and lectotypified specimen of Myelophycus caespitosus. In the genetic analyses using mitochondrial cox1 and cox3, chloroplast atpB, psbA and rbcL DNA sequences, the specimens morphologically referable to M. simplex formed two distinct clades (clade‐1 and clade‐2) supported by high statistical values. Clade‐1 was distributed on the western coast of Japan (Honshu, Shikoku and Kyushu) from the Kii Peninsula to Tsushima and the Pacific coast of Aomori, northeastern Honshu, and clade‐2 on the Pacific coast of central Honshu from Sanriku to the Kii Peninsula. Based on the cox3 DNA sequences and the location of the type locality at Shimoda, Izu Peninsula, clade‐2 was concluded to correspond to true M. simplex. There were no recognizable differences in the representative morphological features (height and diameter of sporophytes and gametophytes, number of cells comprising cortical, subcortical and medullary layers, thickness of plurilocular gametangia, or length and diameter of unilocular zoidangia) between the specimens included in the two taxa. Analyses of the specimens of the two taxa, including sympatric populations on the Kii Peninsula, using a nuclear genetic marker ocm3 did not suggest any genetic exchanges between the two taxa. On the other hand, cox3 gene sequence of the voucher specimen of Myelophycus caespitosus collected from Goto, Kyushu, western Japan housed in the UPS herbarium was included in clade‐1. In conclusion, we propose the reinstatement of M. caespitosus and to lectotypify the specimen in UPS. Korean Myelophycus specimens reported from Cheju Island and Wando were considered to belong to M. caespitosus based on the reported rbcL sequences.     

Egg-feeding in the freshwater piscicolid leech Cystobranchus virginicus (Annelida, Hirudinea)

Abstract. Leeches are common terrestrial and aquatic annelids, parasitizing or feeding on a wide range of host taxa. Here we report evidence for an unusual feeding behavior of egg‐feeding in the piscicolid leech Cystobranchus virginicus. We identified distended specimens of C. virginicus in the nests of at least 4 different fish species: Campostoma anomalum, Moxostoma carinatum, Moxostoma sp. (either M. anisurum and/or M. breviceps), and Nocomis leptocephalus. We collected a total of 41 leeches from the nests of these host species and documented at least 1 leech in 19 of 55 nests (35%), with many sites containing multiple leeches. Individuals of C. virginicus were not identified feeding on any of the 41 adult specimens of Moxostoma spp. or the 635 adult specimens of Nocomis leptocephalus examined, and were never found in the absence of active host spawning (26 sites). These results are consistent with individuals of C. virginicus being an opportunistic or possibly even an obligate egg‐feeder, potentially timing their own reproductive activities with the spawning of their fish hosts. The current distribution of C. virginicus has been expanded to include North Carolina, South Carolina, and Tennessee. The potential for leech species to induce mortality in developing fish eggs could be a concern for fish conservation and merits further investigation.