First records of three Lepiota species (Agaricales, Basidiomycota) from Ukraine, with notes on a poorly known species, Lepiota subalba

New records of four species of the genus Lepiota (Agaricales, Basidiomycota) are reported from Ukraine. Three species, L. fuscovinacea, L. griseovirens, and L. roseolivida, are recorded in Ukraine for the first time, whereas a poorly known species, L. subalba, earlier known in Ukraine from a few records, is confirmed using molecular identification methods. All species reports are supplemented with original descriptions and drawings based on newly collected material, as well as data on general distribution, habitat, references to new collections and public databases. Original nucleotide sequence of the ITS region of ribosomal DNA obtained from our voucher specimen of L. subalba is provided.

Parasites of Free-Ranging and Captive American Primates: A Systematic Review

The diversity, spread, and evolution of parasites in non-human primates (NHPs) is a relevant issue for human public health as well as for NHPs conservation. Although previous reviews have recorded information on parasites in NHPs (Platyrrhines) in the Americas, the increasing number of recent studies has made these inventories far from complete. Here, we summarize information about parasites recently reported in Platyrrhines, attempting to build on earlier reviews and identify information gaps. A systematic literature search was conducted in PubMed, ISI Web of Science, and Latin American and Caribbean Health Sciences Literature (LILACS), and following the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines. Ninety-three studies were included after the screening process. Records for 20 genera of NHPs, including 90 species were found. Most of the studies were conducted on captive individuals (54.1%), and morphological approaches were the most used for parasite identification. The most commonly collected biological samples were blood and stool, and Protozoa was the most frequent parasite group found. There is still scarce (if any) information on the parasites associated to several Platyrrhine species, especially for free-ranging populations. The use of molecular identification methods can provide important contributions to the field of NHPs parasitology in the near future. Finally, the identification of parasites in NHPs populations will continue to provide relevant information in the context of pervasive habitat loss and fragmentation that should influence both human public health and wildlife conservation strategies.

Isolation and Characterization of Nocardiae Associated with Foaming Coastal Marine Waters

Nocardiosis is an infectious disease caused by Nocardia species that occurs worldwide, albeit more prevalently in tropical/subtropical regions. It can appear as either acute, subacute or as a chronic infection mostly with those with a compromised/weakened immune system. Inhalation of spores and or mycelium fragments is the main transmission route for developing pulmonary nocardiosis. In contrast, cutaneous nocardiosis usually occurs via direct contact. In the subtropical region of the Sunshine Coast in Australia foaming events with thick and persistent and orange-brown color foam have been observed during summer seasons in the near shore marine environments. This study reports the existence of nocardiae in these near shore marine environments by the use of a novel isolation method which used the gas requirements of nocardiae as a selective battery. A total of 32 nocardiae were isolated with the use of this novel method and subsequently conducted molecular identification methods confirmed that the isolates belonged to the genus Nocardia. Twenty-one isolates out of the 32 were closely related to N. nova strains MGA115 and one was related to CBU 09/875, in addition when compared with human pathogenic nocardiae twenty of the isolates were found to be related to N. nova strain JCM 6044. Isolates displayed varied resistance against some of the antibiotics tested when interpretation threshold recommended the Comite de L’Antibiogramme de la Societe Francaise de Microbiologie were used. The highest level of resistance against cefotaxime (n = 27) and ceftriaxone (n = 24). Some of the isolates (n = 6) that displayed resistance to selected antibiotics also possessed potential human pathogenic characteristics such as adherence and translocation through human long epithelial cells as well as displaying phage resistance (n = 26). They might thus present a potential public health risk if frequently encountered through exposure to aerosols generated by the foam as well as direct contact through a wound. Preventative measures to control the growth of nocardiae in such environments such as the control of pollutants, might prevent potential infections that might be caused by these bacteria in humans as well as in marine animals.

DNA Barcoding Identifies Unknown Females and Larvae of Fannia R.-D. (Diptera: Fanniidae) from Carrion Succession Experiment and Case Report

Application of available keys to European Fanniidae did not facilitate unequivocal species identification for third instar larvae and females of Fannia Robineau-Desvoidy, 1830 collected during a study of arthropod succession on pig carrion. To link these samples to known species, we took the advantage of molecular identification methods and compared newly obtained cytochrome oxidase subunit I (COI) barcode sequences against sequences deposited in reference databases. As an outcome of the results obtained, we describe for the first time a third instar larva of Fannia nigra Malloch, 1910 and Fannia pallitibia (Rondani, 1866) and a female of Fannia collini d’Assis-Fonseca, 1966. We provide combinations of characters allowing for discrimination of described insects from other Fanniidae. We provide an update for the key by Rozkošný et al. 1997, which allows differentiation between females of F. collini and other species of Fanniidae. Additionally, we provide a case of a human cadaver discovered in Southern Poland and insect fauna associated with it as the first report of F. nigra larvae developing on a human body.

Antibacterial and Antibiofilm Potential of Sea Anemone (Stichodactyla haddoni)-Isolated Vibrio (V. parahaemolyticus and V. alginolyticus), and Pseudoalteromonas (P. gelatinilytica and P. piscicida) Against Staphylococcus aureus and Pseudomonas aeruginosa

Background: Staphylococcus aureus and Pseudomonas aeruginosa are important human bacterial pathogens, which are resistant to several antibiotics. One of the main causes of their resistance is the ability of biofilm formation. Objectives: The present study aimed to evaluate the antibacterial and antibiofilm activity of the extracts of Vibrio parahaemolyticus, V. alginolyticus, Pseudoalteromonas gelatinilytica, and Pseudoalteromonas piscicida isolated from sea anemone (Stichodactyla haddoni) against S. aureus and P. aeruginosa. Methods: Four isolated bacteria were identified using biochemical and molecular identification methods, and their extracts were obtained by mixing the cell-free supernatants from their old broth culture using ethyl acetate and methanol as the solvents. The agar well-diffusion and micro-dilution methods were also applied to determine the antibacterial activity, minimum bactericidal concentration (MBC), and minimum inhibitory concentration (MIC) of the extracts. The ability of the extracts to inhibit biofilm formation and disrupt the preformed biofilm of the pathogens was attained through crystal violet staining in 96-well microtiter plates. To determine the nature of the extracts, they were exposed to protease enzyme, and the antibiofilm activity was compared with the untreated extracts. Results: The extracts of the four isolated bacteria inhibited bacterial growth and biofilm formation and disrupted the preformed biofilm of S. aureus (MIC = BIC = 600 µg/mL) and P. aeruginosa (MIC = BIC = 300 µg/mL). In addition, the active compounds of the extracts with antibiofilm activities were mainly proteases. Conclusions: According to the results, V. parahaemolyticus, V. alginolyticus, P. gelatinilytica, and P. piscicida had antibacterial and antibiofilm potential against S. aureus and P. aeruginosa, and their extract could also be further analyzed as an alternative to antibiotics.

A diagnostic LAMP assay for the destructive grapevine insect pest, phylloxera (Daktulosphaira vitifoliae)

Grape phylloxera (Daktulosphaira vitifoliae) is a destructive insect pest of grapevines that is highly invasive worldwide, despite strict biosecurity containment measures in place at farm and regional levels. Current phylloxera identification by visual inspection and laboratory-based molecular methods is time-consuming and costly. More rapid and cost-effective methods for identification of this pest would benefit industry, growers, and biosecurity services. Loop mediated isothermal amplification (LAMP) is a new portable technology available for rapid and accurate in-field molecular diagnostics. This study outlines the development of a new LAMP assay to enable the identification of phylloxera specimens. New LAMP primers were developed to specifically amplify phylloxera mitochondrial DNA (5′-COI), which we have shown is effective as a DNA barcode for identification of phylloxera, using LAMP technology. Positive LAMP reactions, containing phylloxera DNA, amplified in less than twelve minutes with an anneal derivative temperature of approximately 79 °C to 80 °C compared to a newly designed synthetic DNA (gBlock) fragment which had an anneal derivative temperature of 82 °C. No LAMP amplification was detected in any of the non-target species tested, i.e. no false-positive identification resulted for these species. We also successfully optimised a non-destructive DNA extraction procedure, HotSHOT “HS6”, for use in the field on phylloxera adults, nymphs and eggs, to retain physical specimens. DNA extracted using this method was also suitable for species and genotype molecular identification methods, such as DNA barcoding, qPCR and microsatellite genotyping. The new LAMP assay provides a novel visual molecular tool for accurate diagnostics of phylloxera in the laboratory and field.

The Molecular Data Organization for Publication (MDOP) R package to aid the upload of data to shared databases

Molecular identification methods, such as DNA barcoding, rely on centralized databases populated with morphologically identified individuals and their referential nucleotide sequence records. As molecular identification approaches have expanded in use to fields such as food fraud, environmental surveys, and border surveillance, there is a need for diverse international data sets. Although central data repositories, like the Barcode of Life Datasystems (BOLD), provided workarounds for formatting data for upload, these workarounds can be taxing on researchers with few resources and limited funding. To address these concerns, we present the Molecular Data Organization for Publication (MDOP) R package to assist researchers in uploading data to public databases. To illustrate the use of these scripts, we use the BOLD system as an example. The main intent of this writing is to assist in the movement of data, from academic, governmental, and other institutional computer systems, to public locations. The movement of these data can then better contribute to the global DNA barcoding initiative and other global molecular data efforts.

Isolation and characterization of probiotics bacteria from curd, pickle and fermented rice and screening of antimicrobial activity

The live supplement of microorganisms though the dietary product is known as the probiotics. These are some special kind of bacteria they are healthy for the host organisms. These bacteria commonly found in food and dietary product. In our study, probiotics were isolated from fermented rice, curd and pickle .Total isolated strains were studied for their characterization, and also strains were studied antibacterial also their antibiotic susceptibility quality. The antibiotic resistance of potential strain was studied using Vancomycin, gentamycin, chloromphenicol, ciprofloxacin and cefataximide. PB1, PB3 shows the good resistance activity to the antibiotics. The curd produced by those strains has good quality and quantity of nutrients. Specially this study is for isolating, identifying, and characterizing the probiotics .Isolated strains were identified by Gram staining, catalase assay, and 3 molecular identification methods; namely, (GTG) 5-PCR fingerprinting, This experiments shows the better anti-pathogen activity, and acceptable antibiotic susceptibility; it implies we can use these probiotics in different purpose in food industry for modern food synthesis, for antimicrobial susceptibility test we are taking amplicin, karamycin, tetracycline, tripsin by using of the antibiotics concluded that various inhibition zones of given sample of probiotics.

Annotated checklist and review of the mosquito species (Diptera: Culicidae) in Sri Lanka

Mosquito borne diseases remains as an importance source of morbidity and mortality in Sri Lanka. To better control vectors which transmit the diseases, updated list of the species present in the county is imperative. It is also vital in documenting the diversity of the family Culicidae. Original records were collected from a literature review to compile a list of the species recorded in Sri Lanka. This work illustrates the updated list of mosquito species in Sri Lanka and their current taxonomic status based on previous studies from 1901 to date. A total of 159 species belonging to 19 genera including sibling species, have been included in the revised mosquito checklist in Sri Lanka. The present work includes 13 species, two genera (Lutzia, Verrallina) and 9 subgenera in subfamily Culicinae, tribe Aedini of genus Aedes (Bruceharrisonius, Collessius, Danielsia, Dendroskusea, Downsiomyia, Fredwardsius, Hulecoeteomyia, Neomelaniconion, Phagomyia) in to the checklist which were not included in the previous mosquito checklist published nearly 26 years ago. However, further work is essential to refine this list and to explore the abundance of new species within the country. Improved morphological and molecular identification methods will sanction the refinement of the mosquito catalog in years to come.