Chitosan Sensitivity of Fungi Isolated from Mango (Mangifera indica L.) with Anthracnose.

In Mexico, the mango crop is affected by anthracnose caused by Colletotrichum species. In the search for environmentally friendly fungicides, chitosan has shown antifungal activity. Therefore, fungal isolates were obtained from plant tissue with anthracnose symptoms from the state of Guerrero in Mexico and identified with the ITS and ß-Tub2 genetic markers. Isolates of the Colletotrichum gloeosporioides complex were again identified with the markers ITS, Act, ß-Tub2, GADPH, CHS-1, CaM, and ApMat. Commercial chitosan (Aldrich, lot # STBF3282V) was characterized, and its antifungal activity was evaluated on the radial growth of the fungal isolates. The isolated anthracnose-causing species were C. chrysophilum, C. fructicola, C. siamense, and C. musae. Other fungi found were Alternaria sp., Alternaria tenuissima, Fusarium sp., Pestalotiopsis sp., Curvularia lunata, Diaporthe pseudomangiferae, and Epicoccum nigrum. Chitosan showed 78% deacetylation degree and a molecular weight of 32 kDa. Most of the Colletotrichum species and the other identified fungi were susceptible to 1 g L-1 chitosan. However, two C. fructicola isolates were less susceptible to chitosan. Although chitosan has antifungal activity, the interactions between species of the Colletotrichum gloeosporioides complex and their effect on chitosan susceptibility should be studied based on genomic changes with molecular evidence.

An account of Colletotrichum species associated with anthracnose of Atractylodes ovata in South Korea based on morphology and molecular data.

Ovate-leaf atractylodes (OLA) (Atractylodes ovata) is a well-known medicinal plant in Korea; its dried rhizome and root extracts are used in herbal medicine. However, anthracnose is a great challenge to the OLA cultivation in South Korea. Colletotrichum spp. is a major group of plant pathogens responsible for anthracnose on a range of economically important hosts. Its occurrence on OLA remains unresolved. To investigate the diversity, morphology, phylogeny, and biology of Colletotrichum spp., 32 fungal isolates were obtained from 30 OLA-affected leaves collected from five different farms, in two regions in South Korea, Mungyeong and Sangju. The phylogenetic analysis with four or five gene loci (ITS, TUB2, ACT, GAPDH, and CHS-1) along with morphology of 26 representative isolates delineated six previously known Colletotrichum species including C. fructicola, C. gloeosporioides sensu stricto (s.s), C. cigarro, C. plurivorum, C. siamense and C. sojae, and one new species, described here as C. ovataense. Amongst these species, C. gloeosporioides s.s. and C. plurivorum were the most prevalent species. A pathogenicity test on the detached leaves revealed that different Colletotrichum species presented a distinct degree of virulence, confirming Koch's postulates. In this study, C. fructicola, C. cigarro, C. plurivorum, C. siamense, and C. sojae were reported from A. ovata for the first time, as the causal agent of ovate-leaf atractylodes anthracnose. Understanding the diversity and biology of the Colletotrichum species population will help in managing this disease.

Multi-genetic Analysis of Colletotrichum spp. Associated with Postharvest Disease of Fruits Anthracnose in Special Region of Yogyakarta, Indonesia.

BACKGROUND AND OBJECTIVE: Postharvest disease caused by Colletotrichum spp. caused major losses. The species of Colletotrichum are difficult to distinguish if only seen from their morphology. This study investigated Colletotrichum isolates associated with tropical fruits anthracnose using multi-genetic analysis and the cross-infection potency of each isolate among tropical fruits. MATERIALS AND METHODS: The fruit samples were collected from markets in the Special Region of Yogyakarta, Indonesia and its surrounding area. The fruits affected by anthracnose subjected to isolation, resulting in 15 isolates. Morphology of colony and conidia then characterized and clustered with UPGMA. The seven representative isolates were selected for molecular identification. The multi-genetic analysis was used by combining ITS, Glyceraldehyde 3-phosphate dehydrogenase (gapdh) and tub2 sequence genes. A cross-infection test was conducted by using selected species from the multi-genetic analysis. RESULTS: Multi-genetic analysis clustered the selected isolates into four species. Isolates from banana, avocado, papaya and citrus belonged to gloeosporioides species complex, including C. siamense, C. asianum and C. gloeosporioides. Isolates from apple, guava, mango and citrus belonged to acutatum species complex, including C. sloanei. The cross-infection test in this study showed that C. siamense could cause anthracnose on banana, apple, citrus and avocado, C. asianum on avocado, papaya, apple and citrus, C. gloeosporioides on citrus and apple, C. sloanei on apple, guava, citrus and papaya. CONCLUSION: The C. siamense, C. asianum, C. gloeosporioides and C. sloanei found associated with tropical fruits anthracnose. The potency of the cross-infection test revealed the board range in the pathogenicity of the Colletotrichum isolates.

Species of Colletotrichum on bamboos from China.

Twenty-seven Colletotrichum isolates associated with asymptomatic tissues of bamboo (Bambusoideae, Gramineae) were isolated from Anhui, Beijing, and Guangxi in China. Based on multilocus (internal transcribed spacer [ITS], glyceraldehyde-3-phosphate dehydrogenase [GAPDH], chitin synthase [CHS], actin [ACT], beta-tubulin [TUB2]) phylogenetic analyses and morphological characteristics, three species were distinguished, including two novel species, C. bambusicola and C. guangxiense, and one known species, C. metake, which is a first report for China. These species have hitherto only been discovered on Bambusoideae, indicating that they probably have host preference.

Conyza bonariensis as an alternative host for Colletotrichum species in Argentina.

AIMS: This study investigated the diversity of Colletotrichum isolates recovered from Conyza bonariensis leaves through the use of morphological characteristics, growth rate, carbon sources utilization and phylogenetic analysis. METHODS AND RESULTS: In all, 30 Colletotrichum isolates recovered from C. bonariensis leaves showing symptoms of disease were included in the present study. Based on the analysis of morphology and sequences, the isolates were distributed into six Colletotrichum species complexes. The concatenated alignment of GAPDH and ITS sequences showed that 20 out of 30 isolates were included in four species complexes which comprise the most important pathogens causing anthracnose in soybean or anthracnose and stalk rot in maize: C. truncatum, C. orchidearum, C. gloeosporioides and C. graminicola. The remaining 10 isolates were included in the C. boninense and C. destructivum species complexes or could not be assigned to any complex with the available information. CONCLUSION: Weeds belonging to genus Conyza are host to soybean and maize potential pathogenic species of Colletotrichum and could have a role as inoculum reservoir for cross contamination in the agroecosystem. SIGNIFICANCE AND IMPACT OF THE STUDY: The combined use of morphological, kinetics and physiological parameters of growth and phylogenetic analysis in Colletotrichum isolates from Conyza leaves allowed the detection of species complexes previously not identified in Argentina.

A rare case of Colletotrichum truncatum keratitis in a young boy with complete healing after medical treatment.

Colletotrichum is a rare fungal pathogen, which is known to cause anthracnose in plants and keratitis or subcutaneous infections in humans. Among the seven Colletotrichum species reported in eye infections, truncatum species is usually virulent with poor visual prognosis even after surgical treatment. Here we report a case of Colletotrichum truncatum keratitis in a young boy with thorn injury that completely resolved with topical natamycin and voriconazole.

Identification and characterization of Colletotrichum species causing apple bitter rot in New York and description of C. noveboracense sp. nov.

Apple bitter rot caused by Colletotrichum species is a growing problem worldwide. Colletotrichum spp. are economically important but taxonomically un-resolved. Identification of Colletotrichum spp. is critical due to potential species-level differences in pathogenicity-related characteristics. A 400-isolate collection from New York apple orchards were morphologically assorted to two groups, C. acutatum species complex (CASC) and C. gloeosporioides species complex (CGSC). A sub-sample of 44 representative isolates, spanning the geographical distribution and apple varieties, were assigned to species based on multi-locus phylogenetic analyses of nrITS, GAPDH and TUB2 for CASC, and ITS, GAPDH, CAL, ACT, TUB2, APN2, ApMat and GS genes for CGSC. The dominant species was C. fioriniae, followed by C. chrysophilum and a novel species, C. noveboracense, described in this study. This study represents the first report of C. chrysophilum and C. noveboracense as pathogens of apple. We assessed the enzyme activity and fungicide sensitivity for isolates identified in New York. All isolates showed amylolytic, cellulolytic and lipolytic, but not proteolytic activity. C. chrysophilum showed the highest cellulase and the lowest lipase activity, while C. noveboracense had the highest amylase activity. Fungicide assays showed that C. fioriniae was sensitive to benzovindiflupyr and thiabendazole, while C. chrysophilum and C. noveboracense were sensitive to fludioxonil, pyraclostrobin and difenoconazole. All species were pathogenic on apple fruit with varying lesion sizes. Our findings of differing pathogenicity-related characteristics among the three species demonstrate the importance of accurate species identification for any downstream investigations of Colletotrichum spp. in major apple growing regions.

rps3 as a Candidate Mitochondrial Gene for the Molecular Identification of Species from the Colletotrichum acutatum Species Complex.

Colletotrichum species form one of the most economically significant groups of pathogenic fungi and lead to significant losses in the production of major crops-in particular, fruits, vegetables, ornamental plants, shrubs, and trees. Members of the genus Colletotrichum cause anthracnose disease in many plants. Due to their considerable variation, these fungi have been widely investigated in genetic studies as model organisms. Here, we report the complete mitochondrial genome sequences of four Colletotrichum species (C. fioriniae, C. lupini, C. salicis, and C. tamarilloi). The reported circular mitogenomes range from 30,020 (C. fioriniae) to 36,554 bp (C. lupini) in size and have identical sets of genes, including 15 protein-coding genes, two ribosomal RNA genes, and 29 tRNA genes. All four mitogenomes are characterized by a rather poor repetitive sequence content with only forward repeat representatives and a low number of microsatellites. The topology of the phylogenetic tree reflects the systematic positions of the studied species, with representatives of each Colletotrichum species complex gathered in one clade. A comparative analysis reveals consistency in the gene composition and order of Colletotrichum mitogenomes, although some highly divergent regions are also identified, like the rps3 gene which appears as a source of potential diagnostic markers for all studied Colletotrichum species.

Identification, pathogenicity and histopathology of Colletotrichum sansevieriae causing anthracnose of Sansevieria trifasciata in Malaysia.

AIMS: To characterize causal pathogen of Sansevieria trifasciata anthracnose through morphology and molecular analysis; to evaluate the host range of the pathogen; and to explicate the infection process by the pathogen histopathologically. METHODS AND RESULTS: Symptomatic leaves of S. trifasciata were collected from five states in Malaysia. The causal pathogen was isolated and identified for the first time in Malaysia as C. sansevieriae based on morphological and multi-gene phylogenetic analyses using ITS, TUB2 and GAPDH sequences. Pathogenicity tests were conducted on different hosts. Colletotrichum sansevieriae was not pathogenic towards S. cylindrica, S. masoniana, Furcraea foetida, Chlorophytum comosum, Aloe vera and Gasteria carinata, confirming the exceptionally high host specificity for a species of Colletotrichum. Histopathology was performed using light microscope and scanning electron microscopy to study the infection process of C. sansevieriae on S. trifasciata. Colonization of host leaves by the pathogen was observed 2 days after inoculation. CONCLUSIONS: Colletotrichum sansevieriae caused anthracnose of S. trifasciata in Malaysia. It is a host-specific pathogen and colonized the host intracellularly. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of C. sansevieriae causing anthracnose of S. trifasciata in Malaysia. The host range test and understanding of the infection process will provide better understanding of the host-pathogen relationship and beneficial for effective disease management.

Identification and characterization of Colletotrichum destructivum causing anthracnose on sunflower.

Sunflower is one of the most economically important oil crops. Recently, sunflower anthracnose caused by Colletotrichum destructivum was reported and suggested to be a potential threat to the quality of oil and edible seeds derived from sunflower in the field and even on the ornamentals in the residential gardens. Colletotrichum destructivum, as the causal agent of sunflower anthracnose, has been rarely studied. In this study, the vegetative growth and sporulation of this fungal species were investigated by assessing the requirements of nutrition and other environmental conditions, such as temperature, ambient pH, and lightness regime. Additionally, the sensitivity of C. destructivum to several fungicides was assessed. The results will provide a baseline for better understanding of the biology and etiology of C. destructivum. This study will be the first reference for a sustainable management strategy according to the occurrence and prevalence of the sunflower anthracnose.

Colletotrichum keratitis: A rare yet important fungal infection of human eyes.

BACKGROUND: Colletotrichum is a rare cause of human infection. Previous reports about Colletotrichum keratitis were limited, and most diagnoses from past reports were based on morphological distinction, which could have led to underestimation of the prevalence of Colletotrichum species. OBJECTIVE: We reported phylogenetic analysis, clinical feature and treatment outcome of molecularly diagnosed Colletotrichum keratitis in our hospital. PATIENTS/METHODS: We recruited 65 patients with culture-proven filamentous fungal keratitis between January 1, 2015 and December 30, 2018. Through molecular sequencing including internal transcribed spacer (ITS) and multi-locus phylogenetic analysis of fungal DNA, seven patients were verified as infected with Colletotrichum species, and their medical records were reviewed to determine the clinical characteristics. RESULTS: Six of seven patients had predisposing factors including trauma (5) and immunosuppressive status (1). Six isolates were initially misidentified as other fungi through morphological identification. ITS sequencing identified the isolates belonged to two species complex (SC): C. truncatum and C. gloeosporioides; multi-locus phylogenetic analysis enabled species identification including C. tropicale (3), C. fructicola (2), C. truncatum (1) and C. fusiforme (1). Five patients with C. gloeosporioides SC responded well to medical treatment and two patients with C truncatum SC underwent evisceration because of either no visual potential or intractable pain. CONCLUSIONS: The molecular approach provides accurate diagnosis and raises epidemiological awareness of Colletotrichum keratitis. Through multi-locus phylogenetic analysis, we report the human infections caused by C. tropicale, C. fructicola and C. fusiforme. We also highlight the different clinical outcomes between C. gloeosporioides SC and C. truncatum SC.

Amorphophallus konjac anthracnose caused by Colletotrichum siamense in China.

AIMS: Amorphophallus konjac is an important commercial crop grown in China because it is the only plant species which is rich in glucomannan concentration. Recently, an outbreak of anthracnose (incidence ranging from 10-15%) was observed in a field survey conducted from June to August 2018. This study aims to identify the causal agent of A. konjac anthracnose. METHODS AND RESULTS: The pathogen was isolated on potato dextrose agar (PDA) medium. The fungal colony on PDA was greyish to dark grey. Conidia were falcate, one-celled and hyaline. Based on the micro-morphological and cultural characteristics, the pathogen was identified as Colletotrichum sp. blast search and phylogenetic analysis of the ITS, GAPDH, CHS1, ACT, CAL and TUB2 genes revealed the pathogen as Colletotrichum siamense. Koch's postulates were conducted on 2-month konjac leaves with conidial suspension. Development of typical anthracnose disease was recorded 5 days after inoculation and the pathogen's identity was confirmed by re-isolation and molecular identification. CONCLUSIONS: Amorphophallus konjac anthracnose was caused by C. siamense in China. SIGNIFICANCE AND IMPACT OF THE STUDY: Identification of causal agent of A. konjac anthracnose will be helpful in designing effective disease control strategies.

Distribution and Pathogenicity of Colletotrichum Species Associated With Mango Anthracnose in Mexico.

Mango anthracnose, caused by Colletotrichum spp., is the most significant disease of mango (Mangifera indica L.) in almost all production areas around the world. In Mexico, mango anthracnose has only been attributed to C. asianum and C. gloeosporioides. The aims of this study were to identify the Colletotrichum species associated with mango anthracnose symptoms in Mexico by phylogenetic inference using the ApMat marker, to determine the distribution of these species, and to test their pathogenicity and virulence on mango fruits. Surveys were carried out from 2010 to 2012 in 59 commercial orchards in the major mango growing states of Mexico, and a total of 118 isolates were obtained from leaves, twigs, and fruits with typical anthracnose symptoms. All isolates were tentatively identified in the C. gloeosporioides species complex based on morphological and cultural characteristics. The Bayesian inference phylogenetic tree generated with Apn2/MAT intergenic spacer sequences of 59 isolates (one per orchard) revealed that C. alienum, C. asianum, C. fructicola, C. siamense, and C. tropicale were associated with symptoms of mango anthracnose. In this study, C. alienum, C. fructicola, C. siamense, and C. tropicale are reported for the first time in association with mango tissues in Mexico. This study represents the first report of C. alienum causing mango anthracnose worldwide. The distribution of Colletotrichum species varied among the mango growing states from Mexico. Chiapas was the only state in which all five species were found. Pathogenicity tests on mango fruit cultivar Manila showed that all Colletotrichum species from this study could induce anthracnose lesions. However, differences in virulence were evident among species. C. siamense and C. asianum were the most virulent, whereas C. alienum and C. fructicola were considered the least virulent species.

Colletoindole A from the Mangrove Plant Endophytic Fungus Colletotrichum tropicale SCSIO 41022.

A new indole derivative colletoindole A (1), along with two new indole derivatives (2 and 3) and one known compound acropyrone (4) were isolated from cultures of Colletotrichum tropicale SCSIO 41022 derived from a mangrove plant Kandelia candel. The structures of 1-4 were determined by analysis of NMR and MS data. The cytotoxicity of 1, 2 and 4, and the COX-2 inhibitory activity of 1 and 2 were evaluated.

Optimal markers for the identification of Colletotrichum species.

Colletotrichum is among the most important genera of fungal plant pathogens. Molecular phylogenetic studies over the last decade have resulted in a much better understanding of the evolutionary relationships and species boundaries within the genus. There are now approximately 200 species accepted, most of which are distributed among 13 species complexes. Given their prominence on agricultural crops around the world, rapid identification of a large collection of Colletotrichum isolates is routinely needed by plant pathologists, regulatory officials, and fungal biologists. However, there is no agreement on the best molecular markers to discriminate species in each species complex. Here we calculate the barcode gap distance and intra/inter-specific distance overlap to evaluate each of the most commonly applied molecular markers for their utility as a barcode for species identification. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), histone-3 (HIS3), DNA lyase (APN2), intergenic spacer between DNA lyase and the mating-type locus MAT1-2-1 (APN2/MAT-IGS), and intergenic spacer between GAPDH and a hypothetical protein (GAP2-IGS) have the properties of good barcodes, whereas sequences of actin (ACT), chitin synthase (CHS-1) and nuclear rDNA internal transcribed spacers (nrITS) are not able to distinguish most species. Finally, we assessed the utility of these markers for phylogenetic studies using phylogenetic informativeness profiling, the genealogical sorting index (GSI), and Bayesian concordance analyses (BCA). Although GAPDH, HIS3 and ß-tubulin (TUB2) were frequently among the best markers, there was not a single set of markers that were best for all species complexes. Eliminating markers with low phylogenetic signal tends to decrease uncertainty in the topology, regardless of species complex, and leads to a larger proportion of markers that support each lineage in the Bayesian concordance analyses. Finally, we reconstruct the phylogeny of each species complex using a minimal set of phylogenetic markers with the strongest phylogenetic signal and find the majority of species are strongly supported as monophyletic.

Colletotrichum shisoi sp. nov., an anthracnose pathogen of Perilla frutescens in Japan: molecular phylogenetic, morphological and genomic evidence.

Species of the fungal genus Colletotrichum are among the most devastating pathogens of agricultural crops in the world. Based on DNA sequence data (ITS, GAPDH, CHS-1, ACT, TUB2) and morphology, we revealed Colletotrichum isolates infecting the oil crop Perilla frutescens, commonly known as shiso, to represent a previously unknown species of the C. destructivum species complex and described it as C. shisoi. We found that C. shisoi appears to be able to adopt a hemibiotrophic lifestyle, characterised by the formation of biotrophic hyphae followed by severe necrotic lesions on P. frutescens, but is less virulent on Arabidopsis, compared to its close relative C. higginsianum which also belongs to the C. destructivum species complex. The genome of C. shisoi was sequenced, annotated and its predicted proteome compared with four other Colletotrichum species. The predicted proteomes of C. shisoi and C. higginsianum, share many candidate effectors, which are small, secreted proteins that may contribute to infection. Interestingly, C. destructivum species complex-specific secreted proteins showed evidence of increased diversifying selection which may be related to their host specificities.

Characterization of Colletotrichum Species Causing Anthracnose of Pomegranate in the Southeastern United States.

Anthracnose fruit rot and leaf blight caused by Colletotrichum species are important diseases of pomegranate in the southeastern United States. In this study, 26 isolates from pomegranate were identified based on pathological and molecular characterization. Isolates were identified to species based on multilocus sequence analysis with the internal transcribed spacer region, glyceraldehyde-3-phosphate dehydrogenase, ß-tubulin, and chitin synthase genomic genes. Pomegranate isolates grouped within the C. acutatum and C. gloeosporioides species complexes, with more than 73% belonging to the latter group. Three species were identified within the C. acutatum species complex (C. nymphaeae [n = 5], C. fioriniae [n = 1], and C. simmondsii [n = 1]), and three other species were identified within the C. gloeosporioides species complex (C. theobromicola [n = 11], C. siamense [n = 6], and C. gloeosporioides [n = 2]). Inoculations of pomegranate fruit showed that isolates from the C. acutatum species complex were more aggressive than isolates from the C. gloeosporioides species complex. Interestingly, opposite results were observed when leaves of rooted pomegranate cuttings were inoculated. In addition, Colletotrichum isolates from pomegranate, strawberry, blueberry, mango, and citrus were cross-pathogenic when inoculated to fruit. This is the first study identifying six different species of Colletotrichum causing pomegranate leaf blight and fruit anthracnose in the southeastern United States and the potential cross-pathogenic capability of pomegranate isolates to other commercially important crops.

Detection of Colletotrichum gloeosporioides in native cashew species in Brazil.

The savanna cashew (Anacardium humile A. St. Hil.) nut is a native shrub species distributed in the savanna biome of Central Western Brazil. This species is similar to the cashew (Anacardium occidentale L.) nut, and its edible pseudo-fruits have been used for various purposes. However, A. humile is at risk of extinction. One reason for this is the phytosanitary issues. Therefore, phytosanitary studies of the serious plant diseases caused by fungal pathogens are necessary. The aim of this study was to survey the savanna cashew nut (Anacardium humile) in Central Western Brazil from August 2013 to October 2014. Serious damage caused by anthracnose to the plant's leaves and fruits was consistently detected. The pathogen fungal species was Colletotrichum gloeosporioides. It was identified by morphological characteristics and molecular analyses. Additional experiments showed evidence that C. gloeosporioides can be transmitted by cashew seeds. We observed anthracnose symptoms on the surfaces of the floral peduncle, leaves, and peduncle of the A. humile cashew nuts. The genomic DNA sequences of the selected strains for molecular characterization have had 99% identity with the analogous sequences of C. gloeosporioides. Naturally occurring C. gloeosporioides infection of the leaves of A. humile was low (20.3%) compared with that of the nuts (79.7%). Our results can be used to design strategies to prevent the introduction and establishment of C. gloeosporioides in new areas. They can also be helpful in monitoring programs in areas with a current occurrence of C. gloeosporioides. Finally, these results can be used in future research plans of C. gloeosporioides infection management.

Diversity, Prevalence, and Virulence of Colletotrichum Species Associated with Lima Bean in Brazil.

Anthracnose is one of the most important diseases of lima bean in Brazil. Previously, the disease was attributed exclusively to Colletotrichum truncatum. Therefore, this work aimed to characterize the diversity, prevalence, and virulence of Colletotrichum spp. associated with anthracnose in lima bean in Brazil. Here, we report the species C. truncatum, C. brevisporum, C. lobatum, C. plurivorum, and C. musicola in association with anthracnose of lima bean. All species were pathogenic to lima bean. In addition, several strains were found that represent novel lineages, presented here as Colletotrichum lineages 1 to 5. C. truncatum is the prevailing species and more virulent than all other species studied.

Fungal Keratitis Caused by Colletotrichum dematium: Case Study and Review.

Colletotrichum species are known as important pathogens of plants with an impact on crop production. Some of these species are also known as a cause of rare ophthalmic infections in humans. A case of keratitis caused by Colletotrichum dematium after corneal trauma in a 56-year-old woman is presented. Infection was diagnosed based on positive microscopy and culture. The fungal isolate was identified by morphological characteristics and DNA sequencing of the ITS rDNA region, ß-tubulin (tub2) and glyceraldehyde-3-phosphate dehydrogenase (gapdh) genes. The patient responded well to topical therapy with amphotericin B combined with intravenous amphotericin B but improvement was associated with the corneal collagen cross-linking. The review of the literature revealed another 13 cases of C. dematium keratitis, all but one patient having at least one keratitis risk factor in their history. Almost all patients (n = 12) were treated with topical polyene antibiotics (natamycin or amphotericin B), improvement and cure were achieved in eight of them.