Development of SCAR Markers for Genetic Authentication of Metarhizium acridum.

In this study, molecular typing using Randomly Amplified Polymorphic DNA (RAPD-PCR) was conducted on 16 original isolates of Metarhizium acridum obtained from locusts (Schistocerca piceifrons ssp. piceifrons.) in Mexico (MX). The analysis included reference strains of the genus Metarhizium sourced from various geographical regions. The isolates were identified by phenotypic (macro and micromorphology) and genotypic methods (RAPD-PCR and Amplified Fragment Length Polymorphisms (AFLP), through a multidimensional analysis of principal coordinates (PCoA) and a minimum spanning network (MST). Subsequently, Sequences-Characterized Amplified Region (SCAR) markers were developed for the molecular detection of M. acridum, these markers were chosen from polymorphic patterns obtained with 14 primers via RAPD-PCR. Phenotypic and genotypic characterization identified the MX isolates as M. acridum. Of all the polymorphic patterns obtained, only OPA04 and OPA05 were chosen, which presented species-specific bands for M. acridum, and further utilized to create SCAR markers through cloning and sequencing of the specific bands. The specificity of these two markers was confirmed via Southern hybridization. The SCAR markers (Ma-160OPA-05 and Ma-151OPA-04) exhibit remarkable sensitivity, detecting down to less than 0.1 ng, as well as high specificity, as evidenced by their inability to cross-amplify or generate amplification with DNAs from other strains of Metarhizium (as Metarhizium anisopliae) or different genera of entomopathogenic fungi (Cordyceps fumosorosea and Akanthomyces lecanii). These SCAR markers yield readily detectable results, showcasing high reproducibility. They serve as a valuable tool, especially in field applications.

A review of described cases of mycotic keratitis and sclerokeratitis related to entomopathogenic fungi from 1984 to 2021.

Infectious keratitis and sclerokeratitis caused by filamentous fungi prevail in agricultural regions with tropical and subtropical climates and are related mostly to mild abrasive corneal trauma especially after vegetable matter related injury. Biotechnological advances have introduced biological control agents in agriculture such as fungal-based biocontrol agents that use Beauveria and Metarhizium species as bioinsecticides. Keratitis and sclerokeratitis are the most frequent pathologies associated to Beauveria and Metarhizium infection that are the main entomopathogenic fungi used in biological control, although other clinical cases such as sinus, skin lesions, and disseminated infections have been reported. Search of publications was carried out using the databases: Scopus, Pubmed, ScienceDirect, MedLine Scielo. A total of 30 articles were retrieved from 1984 - 2021. From these, 17 keratitis and one sclerokeratitis clinical cases were related to Beauveria infection, while Metarhizium was linked to 13 keratitis cases and two sclerokeratitis clinical cases. Female sex predominated in both Metarhizium and Beauveria clinical cases, there was no significant difference in sclerokeratitis / keratitis by sex. Contact lenses use was a factor reported in 66.6% cases of infection with Metarhizium and 22.2% with Beauveria. The review of clinical cases of keratitis and sclerokeratitis related to Beauveria and Metarhizium suggests the need to consider entomopathogenic fungi in ocular pathologies and the risk that imply the misuse of contact lenses and agricultural/gardening activities.

Aerobiological study of bacterial and fungal community composition in the atmosphere of Mexico City throughout an annual cycle.

The atmosphere as a temporary habitat for airborne microbial communities is a valuable topic to explore, and it is through aerobiological studies that the diversity of biological particles and their release, emission, transport, deposition, and impact are assessed. Specific microorganisms are involved in meteorological processes, and phytosanitary and public health concerns. Airborne microbial composition is related to factors such as geographic region and weather conditions. In this study a metagenomic approach was used to determine the composition of bacterial and fungal communities in the air of two different land-use areas (urban area and semi-rural area), during dry and rainy seasons in Mexico City. Air sampling was carried out with a Hirst-type spore trap, collecting the samples simultaneously in both study areas. Forty-two bioaerosol samples were collected, and the DNA obtained was sequenced using Next-Generation Sequencing. The results indicated that the bacterial communities were represented mainly by the phyla Actinobacteria, Proteobacteria, Firmicutes, Bacteroidetes, Cyanobacteria, and the fungal communities by the phyla Ascomycota followed by Basidiomycota. The evident changes in microbial composition were related more to seasonality than to locality, since both UA and SRA showed a high degree of urbanization, despite some differences in land use. Continuous monitoring of atmospheric bioaerosols is essential to determine the influence of meteorological factors on the composition of the aerial microbiota.

Sporotrichosis in Mexico.

Sporotrichosis is an endemic mycosis caused by the species of the Sporothrix genus, and it is considered one of the most frequent subcutaneous mycoses in Mexico. This mycosis has become a relevant fungal infection in the last two decades. Today, much is known of its epidemiology and distribution, and its taxonomy has undergone revisions. New clinical species have been identified and classified through molecular tools, and they now include Sporothrix schenckii sensu stricto, Sporothrix brasiliensis, Sporothrix globosa, and Sporothrix luriei. In this article, we present a systematic review of sporotrichosis in Mexico that analyzes its epidemiology, geographic distribution, and diagnosis. The results show that the most common clinical presentation of sporotrichosis in Mexico is the lymphocutaneous form, with a higher incidence in the 0-15 age range, mainly in males, and for which trauma with plants is the most frequent source of infection. In Mexico, the laboratory diagnosis of sporotrichosis is mainly carried out using conventional methods, but in recent years, several researchers have used molecular methods to identify the Sporothrix species. The treatment of choice depends mainly on the clinical form of the disease, the host's immunological status, and the species of Sporothrix involved. Despite the significance of this mycosis in Mexico, public information about sporotrichosis is scarce, and it is not considered reportable according to Mexico's epidemiological national system, the "Sistema Nacional de Vigilancia Epidemiológica." Due to the lack of data in Mexico regarding the epidemiology of this disease, we present a systematic review of sporotrichosis in Mexico, between 1914 and 2019, that analyzes its epidemiology, geographic distribution, and diagnosis.

Biosafety of an entomopathogenic fungus Isaria fumosorosea in an acute dermal test in rabbits.

Isaria fumosorosea (formerly Paecilomyces fumosoroseus) is an entomopathogenic fungus that has been proposed as a low risk environmental alternative to the use of chemical insecticides to control agricultural pests and disease vectors. Although there are some mycoinsecticides already being marketed in many countries, not all their biosafety protocols have been published. The acute dermal toxicity test in an animal model is one in a series of biosafety protocols that must be developed, in order to provide information on health hazards, while taking into consideration the periods that the workers are in direct contact with the microbial agent when applied for the control of pests. For this test, we used I. fumosorosea monospore culture EH-506/3, isolated in Mexico from the Bemisia tabaci whitefly, applying a dose of 2 g/kg of animal body weight, on the shaved skin of 16 New Zealand rabbits, with an exposure time of 24 h. Clinical observations were conducted to evaluate the presence of erythema, edema and other alterations in the skin, as well as the behavior and health of the animals, for a period of 14 days. None of the rabbits showed clinical signs of any disease and their body weight corresponded to the expected weight for a healthy rabbit. The test showed no inflammatory reactions in the skin, supporting the safety of a single dose of this fungus in dermal exposure. Therefore, these data support the safety of I. fumosorosea EH-506/3 when applied to the skin.