Obtaining monokaryotic and dikaryotic mycelial cultures of two Amazonian strains of Geastrum (Geastraceae, Basidiomycota)

The high diversity of the genus Geastrum and the difficulty of obtaining mycelial cultures impairs the study of the ecophysiology and the exploration of the biotechnological potential of the taxon. In this study, different culture media were tested to obtain mycelial cultures for G. lloydianum and G. subiculosum collected in the Brazilian Amazon. Data on spore germination, and isolation of monokaryotic cultures and in vitro sexual reproduction are presented, as well as a brief morphological description of the cultures obtained. For both species, Potato Dextrose Agar (PDA) was the most promising of the tested culture media. The highest growth in agar culture ever recorded for this genus is reported (4.9 mm per week for G. lloydianum and 7.5 mm for G. subiculosum). In the PDA culture medium, spores germinated after 35-40 days of incubation and the isolation of monokaryotic cultures of the two species, as well as in vitro sexual crosses, were successfully performed. (AU)

Research Progress on Forensic Palynology and Its Application in Forensic Science / 法医学杂志

Forensic palynology is to apply palynology to the field of forensic science, using pollen and spores to solve issues in juridical practice, such as civil and criminal issues. Sporopollens have a small size, wide distribution, diverse morphology, can be easily transferred, have durability, and is not easily noticed. It can provide strong investigation and related evidence for case detection as good trace evidence. The application of palynology in forensic science has achieved certain success, but it is underutilized in most countries. This paper analyzes the evidence value provided by sporopollen, collection of the sporopollen samples, the progress in detection technology and challenges ahead, based on the biological characteristics of sporopollen, combined with recent successful cases in forensic science, to comprehensively discuss the research progress in forensic palynology and its application prospects in forensic science.

Tobacco Growth Promotion by the Entomopathogenic Fungus, Isaria javanica pf185

Isaria javanica pf185 is an important entomopathogenic fungus with potential for use as an agricultural biocontrol agent. However, the effect of I. javanica pf185 on plant growth is unknown. Enhanced tobacco growth was observed when tobacco roots were exposed to spores, cultures, and fungal cell-free culture supernatants of this fungus. Tobacco seedlings were also exposed to the volatiles of I. javanica pf185 in vitro using I-plates in which the plant and fungus were growing in separate compartments connected only by air space. The length and weight of seedlings, content of leaf chlorophyll, and number of root branches were significantly increased by the fungal volatiles. Heptane, 3-hexanone, 2,4-dimethylhexane, and 2-nonanone were detected, by solid-phase micro-extraction and gas chromatography-mass spectrophotometry, as the key volatile compounds produced by I. javanica pf185. These findings illustrate that I. javanica pf185 can be used to promote plant growth, and also as a biocontrol agent of insect and plant diseases. Further studies are necessary to elucidate the mechanisms by which I. javanica pf185 promotes plant growth.

The prevalence of allergic diseases in school children of metropolitan city in Indonesia shows a similar pattern to that of developed countries

BACKGROUND: The prevalence of allergy among Surabaya school children is currently unknown. OBJECTIVE: To identify the prevalence of the common allergic sensitization and allergic diseases among school children and undergraduate students in suburban of Surabaya by epidemiologic data collection. METHODS: A multistage simple random sampling was done to select 5 primary schools, 8 secondary schools (4 of junior high schools and senior high schools, respectively), and 1 university from 5 districts in Surabaya city. Out of 550 invited respondents, 499 (128 primary school, 221 secondary school, and 150 undergraduate) respondents gave their consent. A complete personal history, allergic symptoms, environmental exposure of common allergens was obtained from interview and the physical examinations were performed. Skin prick test (SPT) was done using 45 different allergen extracts. Total serum IgE and specific IgE radioallergosorbent test levels were measured for respondents with allergic manifestations. RESULTS: There was an increasing SPT positivity among study respondents, from primary school, secondary school, to undergraduate students (21.90%, 28.95%, to 45.30% respectively). Cockroach (42.85%) and fungi/mold spore (42.85%) were the most common allergens in primary school children. House dust mites was the most common allergen in secondary school (63.16%) and undergraduate students (58.82%). Urticaria and rhinitis were the commonest allergic diseases manifestation. History of atopy was positive in 60.79% of the allergic respondents. CONCLUSION: The prevalence of allergic sensitization among school children and undergraduate students in Surabaya suburb areas were increased compared to previous estimates in 1998. While house dust mites are known as important allergens, surprisingly cockroach was the common allergen among the younger school children.

Evaluation of in vitro antimicrobial susceptibility of Bacillus anthracis strains isolated during anthrax outbreaks in Italy from 1984 to 2017

Anthrax, caused by Bacillus anthracis, is a non-contagious infectious disease that affects a wide range of animal species (primarily ruminants) including humans. Due to the often-fatal outcome in humans, quick administration of definitely effective antimicrobials is crucial either as prophylaxis or as a clinical case therapy. In this study, 110 B. anthracis strains, temporally, geographically, and genetically different, isolated during anthrax outbreaks in Italy from 1984 to 2017, were screened using a broth microdilution method to determine their susceptibility to 16 clinically relevant antimicrobial agents. The strains were isolated from various matrices (human, animal, and environmental samples) and were representative of thirty distinct genotypes previously identified by 15-loci multiple-locus variable-number of tandem repeats analysis. The antimicrobials tested were gentamicin, ceftriaxone, streptomycin, penicillin G, clindamycin, chloramphenicol, vancomycin, linezolid, cefotaxime, tetracycline, erythromycin, rifampin, amoxicillin, ciprofloxacin, doxycycline, and trimethoprim. All isolates were susceptible to most of the tested antimicrobials, with the exception of trimethoprim for which all of them showed high minimal inhibitory concentration values. An intermediate level of susceptibility was recorded for ceftriaxone and cefotaxime. Although the Centers for Disease Control and Prevention recommend the use of doxycycline, ciprofloxacin, penicillin G, and amoxicillin for treatment of human cases and for post-exposure prophylaxis to anthrax spores, this study shows a high degree of in vitro susceptibility of B. anthracis to many other antimicrobials, suggesting the possibility of an alternative choice for prophylaxis and therapy.

Kudoa ogawai (Myxosporea: Kudoidae) Infection in Cultured Olive Flounder Paralichthys olivaceus

Since Kudoa septempuntata was identified as a causative agent of food poisoning associated with raw olive flounder Paralichthys olivaceus, interest and concern regarding the parasite have increased. However, there have been no investigations or reports of other Kudoa species infecting the fish (except for K. paralichthys, which infects the brain) in Korea. We found cysts filled with myxospores of Kudoa species in muscles of cultured olive flounder specimens and identified these to the species level. Mature spores were quadrate, measuring 8.7±0.5 μm in length, 9.2±0.4 μm in thickness, and 12.9±0.6 μm in width. The spores containing 4 polar capsules had a length of 2.1±0.2 μm and a width of 1.8±0.3 μm. The partial 18S and 28S rDNA of isolates showed 99–100% similarities with K. ogawai. Using these morphological and molecular analyses, the species was identified as K. ogawai. This study is the first report of K. ogawai infection in cultured olive flounder in Korea.

Sinuolinea capsularis (Myxosporea: Sinuolineidae) Isolated from Urinary Bladder of Cultured Olive Flounder Paralichthys olivaceus

Sinuolinea capsularis Davis, 1917 is myxosporean that infect the urinary system of the host fish. Insufficient morphological and molecular data of S. capsularis exits, and it is therefore difficult to make an accurate identification of the parasite. We tried a series of morphological and molecular analysis to identify an myxosporean isolated from urinary bladder of cultured olive flounder, Paralichthys olivaceus, from Jeju island in the Republic of Korea. Some of them were observed under a light microscope and SEM, and remain samples were used molecular and phylogenetic analysis. Mature spores were subspherical, measuring 13.9±0.6 μm in length and 13.8±0.8 μm in width. Two spherical polar capsules on opposite sides in the middle of the spore had a diameter range of 4.3±0.4 μm. Scanning electron microscopy revealed that spores a severely twisted the suture line. By the morphological comparison and analysis, it was identified as S. capsularis. In addition, we obtained the partial 18S rDNA of S. capsularis and first registered it in NCBI. Phylogenetic analysis showed that S. capsularis clustered with Zschokkella subclade infecting the urinary system of marine fish, and it supported the infection site tropism effect on phylogeny of marine myxosporeans as well as the origin of Sinuolinea is not monophyly.

Chemical composition and antifungal activity of propolis on Aspergillus flavus / Composição química e atividade antifúngica de própolis sobre Aspergillus flavus

Research has indicated the antifungal activity of ethanol extracts from propolis (EEP) on fungi of the genera Phakopsora, Colletotrichum, and Cercospora. Here, chemical compositions and antioxidant activity of three EEP (propolis from Scaptotrigona polysticta stingless bee and two types produced by Apis mellifera - red and brown) were evaluated and their action against Aspergillus flavus was investigated. Ash, dry extract, total phenolic and total flavonoid contents were determined. Phenolic composition was evaluated by high performance liquid chromatography, with using the following reference substances: gallic acid, caffeic acid, -coumaric acid, ferulic acid, quercetin, kaempferol, and apigenin. Tests on mycelial growth, sporulation, and germination of spores were performed to assess the biological activity of the EEP on A. flavus. All EEP showed low dry extract content (<11%) with adequate amounts of ash (<5%). The red EEP (Apis mellifera) showed the highest contents of total phenolic and total flavonoid (5.38 and 2.77 g 100 g-1), while the highest recorded antioxidant activity was exhibited by brown EEP (92.9%). The EEP of S. polysticta presented higher levels of -coumaric acid (10.99 mg g-1), while red and brown extracts from A. mellifera stood out with the highest levels of quercetin (27.26 mg g-1) and gallic acid (5.88 mg g-1), respectively. No extract was effective in inhibiting mycelial growth and sporulation of A. flavus, but red EEP inhibited spore germination. On the basis of these results, it is suggested that the inhibitory effect of red EEP on spore germination may be associated with increased levels of flavonoids found in it as compared with the other extracts investigated here.

Outros estudos apontaram a ação antifúngica dos extratos etanólicos de própolis (EEP) sobre fungos do gênero Phakopsora, Colletotrichum e Cercospora. Neste estudo, avaliou-se a composição química e a atividade antioxidante de três EEP (própolis produzido pela abelha sem ferrão Scaptotrigona polysticta e dois tipos de própolis produzida pela Apis melífera: marrom e vermelha) e sua ação contra o Aspergillus flavus. Foram determinados os teores de cinzas, extrato seco, fenólicos e flavonoides totais nos EEP. A composição fenólica foi determinada por cromatografia líquida de alta eficiência, empregando-se as seguintes substâncias de referência: ácido gálico, ácido p-cumárico, ácido ferrúlico, quercitina, kaempferol e apigenina. Para avaliar a atividade biológica dos EEP sobre A. flavus foram realizados os testes de crescimento micelial, esporulação e germinação de esporos. Todos os EEP apresentaram baixos teores de extrato seco (< 11%), porém quantidades de cinzas adequadas (< 5%). O EEP vermelha apresentou os maiores teores de fenólicos e flavonoides totais (5,38 e 2,77 g 100g-1), enquanto que a maior atividade antioxidante registrada foi do EEP marrom (92,9%). O EEP de S. polysticta apresentou maiores teores de ácido p-cumárico (10,99 mg g-1), enquanto que os de A. mellifera, vermelha e marrom, destacaram-se com os maiores níveis de quercetina (27,26 mg g-1) e de ácido gálico (5,88 mg g-1), respectivamente. Nenhum extrato foi eficaz na inibição do crescimento micelial e esporulação de A. flavus, porém observou-se inibição na germinação de esporos pelo EEP vermelha. Sugere-se que o efeito inibitório na germinação de esporos pode estar relacionado com os maiores níveis de flavonoides encontrados na própolis vermelha, quando comparado com os outros extratos investigados.

Experimental Study to Develop a Method for Improving Sample Collection to Monitor Laryngoscopes after Reprocessing

BACKGROUND/AIMS: The microbiological surveillance of endoscopes and automated flexible endoscope reprocessing have been proven to be two of the most difficult and controversial areas of infection control in endoscopy. The purpose of this study was to standardize a sampling method for assessing the effectiveness of standard reprocessing operating procedures for flexible fiberoptic laryngoscopes (FFLs). METHODS: First, the sampling devices were directly inoculated with Bacillus atrophaeus spores; second, tissue non tissue (TNT) wipes were tested on artificially contaminated surfaces and on FFLs. RESULTS: Comparison of the sponges, cellulose, and TNT wipes indicated that the TNT wipes were more effective in releasing spores (93%) than the sponges (49%) and cellulose wipes (52%). The developed protocol provides a high efficiency for both collection and extraction from the stainless steel surface (87% of the spores were removed and released) and from the FFL (85% of the spores were removed and released), with relatively low standard deviations for recovery efficiency, particularly for the analysis of the FFL. CONCLUSIONS: TNT wipes are more efficient for sampling surface areas, thereby aiding in the accuracy and reproducibility of environmental surveillance.

Review of Disinfection and Sterilization – Back to the Basics

In-depth knowledge of disinfection and sterilization is a key component of infection control. Sterilization completely removes a spore, whereas disinfection cannot. Disinfectants are classified as oxidants and non-oxidants. The decision regarding which method to apply is based on Spaulding's classification. In this article, disinfection and sterilization are thoroughly reviewed, and extensive information from basic to practical points is discussed.

Review of Disinfection and Sterilization – Back to the Basics

In-depth knowledge of disinfection and sterilization is a key component of infection control. Sterilization completely removes a spore, whereas disinfection cannot. Disinfectants are classified as oxidants and non-oxidants. The decision regarding which method to apply is based on Spaulding's classification. In this article, disinfection and sterilization are thoroughly reviewed, and extensive information from basic to practical points is discussed.

Effects of Long-Term Subcultured Arbuscular Mycorrhizal Fungi on Red Pepper Plant Growth and Soil Glomalin Content

Arbuscular mycorrhizal fungi (AMF) are well-known for their ability to improve plant growth and help plants withstand abiotic stress conditions. Unlike other fungi and bacteria, AMF cannot be stored, as they are obligate biotrophs. Long-term preservation of AMF spores is challenging and may lead to the loss of viability and efficiency. This study aimed to understand the effect of prolonged subculture of AMF species on the growth and glomalin-related soil protein (GRSP) from red pepper (Capsicum annuum L.). AMF spores were mass-produced using different techniques and subcultured in pots with sorghum sudangrass as the host plant for 3 years. Experimental soil samples were collected from natural grassland. Five different AMF inocula were used in triplicate as treatments. After 70 days of growth, red pepper plants were harvested and plant dry weight, plant nutrient content, mycorrhizal colonization, AMF spore count, and soil glomalin content were determined. AMF-treated plants displayed higher dry weight than controls, with only fruit dry weight being significantly different. Similarly, significant differences in phosphorous and potassium contents of the above-ground plant parts were observed between mycorrhizal and control treatments. In addition, soil GRSP content was significantly higher in plants inoculated with Rhizophagus sp. and Gigaspora margarita. The increased plant growth and GRSP content suggest that AMF can be maintained for 3 years without losing their efficiency if subcultured regularly with different symbiotic host plants.

Timing of Fusarium Head Blight Infection in Rice by Heading Stage

Fusarium graminearum causes the devastating plant disease Fusarium head blight and produces mycotoxins on small cultivated grains. To investigate the timeframe of F. graminearum infection during rice cultivation, a spore suspension of F. graminearum was applied to the rice cultivars Dongjin 1 and Nampyeongbyeo before and after the heading stage. The disease incidence rate was the highest (50%) directly after heading, when the greatest number of flowers were present, while only 10% of the rice infected 30 days after heading showed symptoms. To understand the mechanism of infection, an F. graminearum strain expressing green fluorescent protein (GFP) was inoculated, and the resulting infections were visually examined. Spores were found in all areas between the glume and inner seed, with the largest amount of GFP detected in the aleurone layer. When the inner part of the rice seed was infected, the pathogen was mainly observed in the embryo. These results suggest that F. graminearum migrates from the anthers to the ovaries and into the seeds during the flowering stage of rice. This study will contribute to uncovering the infection process of this pathogen in rice.

Acaulosproa koreana, a New Species of Arbuscular Mycorrhizal Fungi (Glomeromycota) Associated with Roots of Woody Plants in Korea

A new species of arbuscular mycorrhizal fungi (Glomeromycota), Acaulospora koreana, was isolated from forest soils in South Korea. This novel fungus was collected from the rhizosphere of Lindera obtusiloba and Styrax obassia in forest and propagated with Sorghum bicolor in pot. Morphological characteristics of spores of A. koreana are rarely distinguished from Acaulospora mellea, which is reported as one of the most abundant mycorrhizal species in Korea. However, molecular evidence of rDNA sequence using improved primers for glomeromycotan fungal identification strongly supported that A. koreana is different from A. mellea but also any other species belonging to the genus Acaulospora. This is the first novel glomeromycatan fungus introduced in South Korea, but it suggests that there is a high possibility for discovering new arbuscular mycorrhizal fungi considering the abundance of plant species and advanced phylogenetic analysis technique.

Dual Biocontrol Potential of the Entomopathogenic Fungus, Isaria javanica, for Both Aphids and Plant Fungal Pathogens

Dual biocontrol of both insects and plant pathogens has been reported for certain fungal entomopathogens, including Beauveria bassiana and Lecanicillum spp. In this study, we demonstrate, for the first time, the dual biocontrol potential of two fungal isolates identified by morphological and phylogenetic analyses as Isaria javanica. Both these isolates caused mortality in the greater wax moth, and hence can be considered entomopathogens. Spores of the isolates were also pathogenic to nymphs of the green peach aphid (Myzus persicae), with an LC₅₀ value of 10⁷ spores/mL 4 days after inoculation and an LT₅₀ of 4.2 days with a dose of 10⁸ spores/mL. In vitro antifungal assays also demonstrated a strong inhibitory effect on the growth of two fungi that are pathogenic to peppers, Colletotrichum gloeosporioides and Phytophthora capsici. These results indicate that I. javanica isolates could be used as novel biocontrol agents for the simultaneous control of aphids and fungal diseases, such as anthracnose and Phytophthora blight, in an integrated pest management framework for red pepper.

Clostridium botulinum spores in Polish honey samples

The aim of this study was an examination of 240 multifloral honey samples collected from Polish apiaries to determine Clostridium botulinum occurrence. Honey was collected from apiaries directly after the extraction process. Samples were inoculated by using the dilution and centrifugation method. Suspected isolates were examined by using mouse bioassay, polymerase chain reaction (PCR), and real-time PCR methods. C. botulinum type A and B strains were detected in 5 of 240 examined honey samples (2.1%). Bacterial strains were also detected that were phenotypically similar to C. botulinum but that did not exhibit the ability to produce botulinum toxins and did not show the presence of the botulinum cluster (ntnh and bont genes) or expression of the ntnh gene. The methods used in the examination, especially the expression analysis of ntnh gene, enabled specific analysis of suspected strains and could be used routinely in environmental isolate analyses of C. botulinum occurrence.

Palynological analysis of Dennstaedtiaceae taxa from the Paranaense Phytogeografic Province that produce monolete spores and its systematic implications (I): Blotiella lindeniana, Histiopteris incisa and Paesia glandulosa

ABSTRACT The genera of Dennstaedtiaceae have sporophytes with very different morphological characteristics between each other, and this feature has made difficult the systematic circumscription of the family. This reason makes necessary the study of new characters that allow a better understanding of the relations within the group. The spore morphology and wall structure of Blotiella lindeniana, Histiopteris incisa and Paesia glandulosa from the Paranaense Phytogeographic Province were studied using light microscope, and scanning and transmission electron microscope. The exospore has two layers and, according to the species, the exospore surface bears pila, echinae, verrucae, bacula and tubercles. The perispore has two or three layers and its surface is psilate, baculate or rugulate. The variability found in the sculpture of the spores and their stratification and ultrastructure of perispore reflects the morphological differences observed in the sporophyte of the species studied. Additionally, while the stratification and ultrastructure of the exospore is shared by the Dennstaedtiaceae species, their ornamentation could be a character to distinguish species into the clade "hypolepidoide". The finding of spores with similar characteristics in phylogenetically unrelated families allows us to suggest that palynological features do not have an evolutionary value to establish relationships between groups above the genus level.

ÁNTRAX: enfermedad aún vigente / ANTHRAX: disease still current

El ántrax es una zoonosis producida por Bacillus anthracis, único miembro del género Bacillus que es capaz de causar enfermedad epidémica en humanos y otros mamíferos. Afecta principalmente a los animales herbívoros domésticos y silvestres. Los humanos son hospederos accidentales y se infectan por contacto directo o indirecto con animales o productos contaminados. Las esporas pueden vivir en el suelo por años y los humanos pueden contraer el ántrax al tener contacto con animales infectados, productos provenientes de estos que al consumir carne infectada; esto se presenta principalmente en países poco desarrollados donde los niveles de vacunación animal contra esta enfermedad son bajos. Este escrito tiene por objetivo presentar una revisión sobre el tema, especialmente sobre aspectos como el modo de infección, manifestaciones clínicas, diagnóstico y tratamiento de la enfermedad.

Anthrax is a zoonosis produced by Bacillus anthracis, the only member of the genus Bacillus that is capable of causing epidemic disease in humans and other mammals. It mainly affects wild and domestic animals. Humans are accidental hosts and are infected through direct or indirect contact with animals or contaminated animal products. B. anthracisspores can live in the soil for many years and humans can become infected with anthraxby contact with infected animals or products contaminated from eating meat infected. This disease occurs mainly in developing countries where vaccine levels are low.The objective of this paper is to present a review on thesubject, especially on aspects such as mode ofinfection, clinical features diagnostic assessment and treatment of thedisease.

Is there evidence that Kudoa septempunctata can cause an outbreak of acute food poisoning? / 한국역학회지

After publishing results of a study that revealed diarrheagenic and emetic activity in 4-5-day old mice infected with Kudoa septempunctata (K. septempunctata) spores, the Korea Centers for Disease Control and Prevention reported 11 events of “Kudoa food poisoning” in 2015. The epidemiological design of the previous study was descriptive rather than analytical; therefore, this study aimed to further investigate the pathogenicity of K. septempunctata. Academic articles showing evidence of the pathogenicity of K. septempunctata were searched via PubMed using the citation discovery tool. Information regarding the kinds of experimental animals and inoculum spores used, as well as study results were extracted. Four articles evaluating the pathogenicity of Myxospran parasites were selected; the first article suggested the pathogenicity of K. septempunctata, while the remaining three articles reported no abnormal symptoms or histopathologic changes. Our findings indicate that there is weak evidence supporting the pathogenicity of K. septempunctata. Further studies evaluating the pathogenicity of K. septempunctata are needed urgently.

Development of High Cordycepin-Producing Cordyceps militaris Strains

Cordyceps militaris, known as Dong-Chong-Xia-Cao, produces the most cordycepin among Cordyceps species and can be cultured artificially. For these reasons, C. militaris is widely used as herb or functional food in the East Asia. In this study, we developed a new strain of C. militaris that produces higher cordycepin content than parent strains through mating-based sexual reproduction. Twenty parent strains were collected and identified as C. militaris based on internal trasncrived spacer and rDNA sequences. Seven single spores of MAT 1-1 idiomorph and five single spores of MAT 1-2 idiomorph were isolated from 12 parent strains. When 35 combinations were mated on the brown rice medium with the isolated single spores, eight combinations formed a stroma with a normal perithecia and confirmed mated strains. High pressure liquid chromatography analysis showed that mated strain KSP8 produced the most cordycepin in all the media among all the tested strains. This result showed due to genetic recombination occurring during the sexual reproduction of C. militaris. The development of C. militaris strain with increased cordycepin content by this approach can help not only to generate new C. militaris strains, but also to contribute to the health food or medicine industry.