Bacillus licheniformis M2-7 Decreases Ochratoxin A Concentrations in Coffee Beans During Storage.

Microbial contamination of coffee beans arises from various factors such as harvesting, handling, and storage practices, during which ochratoxin A (OTA)-producing fungi develop and proliferate. The presence of elevated concentrations of OTA poses a serious health risk to coffee consumers. Therefore, the implementation of a post-harvest treatment involving the use of bacteria known to antagonize OTA-producing fungi constitutes a safe alternative for reducing or eliminating the toxin's concentration in coffee beans. In this study, coffee beans (Coffea arabica L.) were inoculated with Bacillus licheniformis M2-7, after which we monitored fungal growth, in vitro antagonism, and OTA concentration. Our findings demonstrated that coffee beans inoculated with this bacterial strain exhibited a significant decrease in fungal populations belonging to the genera Aspergillus and Penicillium, which are known to produce OTA. Moreover, strain M2-7 decreased the growth rates of these fungi from 67.8% to 95.5% (P < 0.05). Similarly, inoculation with B. licheniformis strain M2-7 effectively reduced the OTA concentration from 24.35 ± 1.61 to 5.52 ± 1.69 µg/kg (P < 0.05) in stored coffee beans. These findings suggest that B. licheniformis M2-7 holds promise as a potential post-harvest treatment for coffee beans in storage, as it effectively inhibits the proliferation of OTA-producing fungi and lowers the toxin's concentration.

First Report of Rhizoctonia solani AG-4 HG-I Causing Root and Basal Stem Rot on Roselle (Hibiscus sabdariffa) in Mexico.

Roselle (Hibiscus sabdariffa L.) is a crop of economic importance, refreshing drinks are prepared from its calyces, it is also attributed to antioxidant, antibacterial, and antihypertensive properties (Da-Costa-Rocha et al. 2014). In November 2022, in municipality of Iguala (18.355592N, 99.548546W, 749 m above sea level), Guerrero, México, roselle plants of approximately 1.5 months of age with basal rot were detected under greenhouse conditions. The symptoms consisted of wilting, yellowing, and root and stem rot with constriction in the base of the stem. The symptoms were detected in approximately 15% of plants at the operation. From symptomatic tissue, cuts were made into approximately 0.5 cm pieces, sterilized with 2% NaClO, washed with sterile distilled water, transferred to PDA medium amended with 50 mg/liter of Chloramphenicol, and incubated in the dark for four days at 28 °C. Rhizoctonia-like colonies were consistently obtained, and nine isolates were selected and purified by the hyphal-tip method. After four days, isolates developed a mycelium was light-white that became brown with age. Right-angled hyphal branching was also observed, in addition to a slight constriction at the base of the branches. In some older cultures, numerous dark brown sclerotia were observed. They were multinucleate cell with three to eight nuclei and measured from 1 to 2 mm in diameter. Together these characteristics were consistent with the description of Rhizoctonia solani Kühn (Parmeter 1970). The anastomosis group (AG) was confirmed by amplifying the ITS region with the primers ITS1 and ITS4 (White et al. 1990) of the RIJAM3 and RIJAM5 strains. The sequences were deposited in GenBank (Nos. OR364496 and OR364497 for RIJAM3 and RIJAM5, respectively). BLAST analysis, both isolates indicated 99.7 identity to R. solani AG-4 HG-I (GenBank: KM013470) strain ICMP 20043 (Ireland et al. 2015). The phylogenetic analysis of AGs sequences allowed assignment of isolates RIJAM3 and RIJAM5 to the AG-4 HG-1 clade. A pathogenicity test was performed on 20 one-month-old roselle plants. Mycelium of RIJAM3 isolate was inserted into the base of the stem with a sterile toothpick. As a control, a sterile toothpick with no mycelium was inserted in ten healthy plants. Additionally, 50 eight-day-old seedlings were inoculated by placing a 5-mm diameter agar plug colonized with mycelium of RIJAM3 at the base of the stem 10 mm below the soil surface. As control treatments, uncolonized PDA plugs were deposited at the base of 25 seedlings. The inoculated plants were incubated in a greenhouse with an average temperature and relative humidity of 28°C and 85%, respectively. Following inoculation, symptoms similar to those observed in the original outbreak were observed in plants after six days and only after four days in seedlings. In both experiments, the control plants and seedlings remained asymptomatic. R. solani was re-isolated from plants and seedlings, complying with Koch's postulates. The pathogenicity testing was repeated twice, with concordant results. In Nigeria and Malaysia R. solani was reported to seedling death to cause seedling dieback in roselle (Adeniji 1970; Eslaminejad and Zakaria 2011). In México R. solani AG-4 has been previously reported in crops of potato, chili and tomato (Montero-Tavera et al. 2013; Ortega-Acosta et al. 2022; Virgen-Calleros et al. 2000). To the best of our knowledge, this is the first report of R. solani AG-4 HG-I as a causing of root and basal stem rot on roselle in Mexico. This research provides information essential for informing the management of this disease, and may help design measures to prevent the spread of the pathogen to other regions.

The catE gene of Bacillus licheniformis M2-7 is essential for growth in benzopyrene, and its expression is regulated by the Csr system.

Benzopyrene is a high-molecular-weight polycyclic aromatic hydrocarbon that is highly recalcitrant and induces carcinogenic effects. CsrA is a conserved regulatory protein that controls the translation and stability of its target transcripts, having negative or positive effects depending on the target mRNAs. It is known that Bacillus licheniformis M2-7 has the ability to grow and survive in certain concentrations of hydrocarbons such as benzopyrene, prompted in part by CsrA, as is present in gasoline. However, there are a few studies that reveal the genes involved in that process. To identify the genes involved in the Bacillus licheniformis M2-7 degradation pathway, the plasmid pCAT-sp containing a mutation in the catE gene was constructed and used to transform B. licheniformis M2-7 and generate a CAT1 strain. We determined the capacity of the mutant B. licheniformis (CAT1) to grow in the presence of glucose or benzopyrene as a carbon source. We observed that the CAT1 strain presented increased growth in the presence of glucose but a statistically considerable decrease in the presence of benzopyrene compared with the wild-type parental strain. Additionally, we demonstrated that the Csr system positively regulates its expression since it was observed that the expression of the gene in the mutant strain LYA12 (M2-7 csrA:: Sp, SpR) was considerably lower than that in the wild-type strain. We were thus able to propose a putative regulation model for catE gene in B. licheniformis M2-7 strain by CsrA regulator in the presence of benzopyrene.

First report of Lasiodiplodia theobromae causing leaf rot on Agave angustifolia in Mexico.

The agave crop (Agave angustifolia), is of economic importance for Mexico, for the agave is made mainly an alcoholic beverage called locally mezcal. In the state of Guerrero, in the municipality of Huitzuco de los Figueroa (18.2510026N, 99.2320182W, 1196 m above sea level), a severe disease affecting agave leaves was detected. The field symptoms consisted of pale to brown dark descending lesions, covering >50% of the leaf surface, in which the presence of pycnidia was observed. In an estimated area of 0.5 ha, the estimated incidence was 67% (n=100 plants). Symptomatic fragments from leaves (approximately 0.5 cm) were taken, superficially disinfected with 1% NaClO, and rinsed twice with sterile distilled water. Then they were transferred to potato dextrose agar (PDA) medium, and incubated at 28 °C. After five days, twelve representative isolates were selected and purified by the hyphal tip technique. In the PDA medium, the colonies were initially light gray, later they became dark, and after 22 days of incubation, the development of numerous dark pycnidia was observed on the surface of the medium. Initially, immature hyaline conidia, unicellular, oval, and double-walled were observed. The mature conidia were dark brown, oval, with one septum and longitudinal striation, and measured 17.5 to 27 [average 25.3 µm; n=50] × 10.5 to 15.7 [average 13.9 µm; n=50]. Based on the morphological characteristics, the fungus was identified as Lasiodiplodia theobromae (Pat.) Griffon & Maubl. (Alves et al. 2008). Isolates LAS3 and LAS4 were used for molecular identification, this was done by amplifying the regio internal transcribed spacer (ITS) of rDNA with primers ITS1 and ITS4 (White et al. 1990) and translation elongation factor 1-alpha ( EF-1α) genes using primers EF1-728F/EF1-986R (Carbone and Kohn 1999). The resulting sequences were deposited in GenBank (LAS3; ON391564 and LAS4; ON391565 for ITS, and LAS3; ON368190 and LAS4; ON368191 for EF-1α). BLASTn analysis sequences of isolated LAS3 and LAS4 revealed for ITS 98.6% identity with L. theobromae (MK934699.1), and for EF-1α indicated 100% identity (MF422024.1). From concatenated sequences ITS-EF-1α regions, a phylogenetic analysis was carried out in MEGA X software, using the Maximum Likelihood and Kimura 2-parameter model with 1,000 bootstraps replicated; isolates LAS3 and LAS4 were clustered in the clade of the members of L. theobromae strains CAA006 (Alves et al. 2006), and INTA-IMC 1601 (Perez et al. 2018). The pathogenicity tests were carried out on 10 healthy 3 year-old agave plants, in which the mycelium of the LAS4 isolate was inserted at three equidistant points/leaf, using a sterile toothpick. Five healthy agave plants were inoculated only with sterile PDA as control treatment. The inoculated plants were covered with transparent plastic bags and housed in a greenhouse at 28 °C. After seven days, similar symptoms to those observed in the field were observed in all inoculated plants. Control plants did not develop symptoms. The fungus L. theobromae was re-isolated again from the infected leaves, fulfilling Koch's postulates. In China, L. theobromae has been reported as the cause of leaf rot on A. sisalana (Xie et al. 2016). To our knowledge, this is the first report of L. theobromae causing leaf rot on A. angustifolia in Mexico. This research is useful to design management strategies for leaf rot disease for local farmers of A. angustifolia.

Antagonic and plant growth-promoting effects of bacteria isolated from mine tailings at El Fraile, Mexico.

Mine tailings contain high concentrations of heavy metals such as As, Pb, Cu, Mn, and Fe, which are detrimental to the health of humans and the environment. In tailings at the El Fraile mine in Guerrero, Mexico, some plant species are apparently tolerant of heavy metals and can be found growing in the tailings. These plants could be associating with heavy metal-tolerant bacteria that promote plant growth and improve biomass production, and these bacteria could be a useful alternative for bacteria-assisted phytoremediation. The objective of this study was to isolate bacteria detected in the mine tailings at El Fraile-Taxco, focusing on those in the soil from the rhizosphere, the inner tissue of the root, leachate, and water, which have the potential to promote plant growth. The ability of the isolated bacteria to promote plant growth was evaluated in vitro. Of the 151 morphotypes isolated, 51% fix nitrogen, 12% dissolve phosphates, and 12%, 39.7%, and 48.3% produce indole acetic acid, gibberellins, and siderophores, respectively. In addition, 66.7% were observed to produce lytic enzymes, such as proteases, celluloses, lipases, esterases, and amylases, which exhibited activity against Fusarium, Aspergillus, and Colletotrichum. The use of 16S rRNA analysis led to the identification of the bacterial genera Chryseobacterium, Bacillus, Pseudomonas, Mycobacterium, Staphylococcus, Curtobacterium, Enterobacter, Agrobacterium, Ochrobactrum, Serratia, Stenotrophomonas, and Acinetobacter. The bacteria isolated from the rhizosphere exhibited the greatest ability to fix nitrogen and produced indole acetic acid, gibberellins, siderophore, and lytic enzymes. In addition, the isolates collected from the soil samples demonstrated ability to solubilize phosphate.

A Review of Biological Control One Decade After the Sorghum Aphid (Melanaphis sorghi) Outbreak.

Melanaphis sorghi is a pest that is native to Africa but is now distributed worldwide. In 2013, its destructive capacity was demonstrated when it devastated sorghum crops in the United States and Mexico, making it a new pest of economic importance in North America. At the time, the phytosanitary authorities of both countries recommended the use of pesticides to control the outbreak, and biological control products for the management of this pest were not known. In response to the outbreak of M. sorghi in North America, several field studies have been performed in the last decade on sorghum crops in the USA and Mexico. Works have focused on assessing resistant sorghum hybrids, pesticide use, and recruitment of associated aphid predators and entomopathogens for natural control of M. sorghi populations. The objective of this review is to compile the information that has been generated in the past decade about indigenous enemies affecting M. sorghi naturally in the field, as well as the search for biological control alternatives and evaluations of interactive effects of resistant sorghum hybrids, pesticides, and natural enemies. To date, different predators, parasitoids, fungi, and bacteria have been evaluated and in many cases found to affect M. sorghi populations in sorghum agroecosystems or laboratory bioassays, and the use of resistant sorghum varieties and pesticides did not have clear toxic effects on natural enemy populations. Many of the macroorganisms and microorganisms that have been evaluated as potential biological controls have shown potential as alternatives to synthetic pesticides for keeping M. sorghi population densities below economic damage thresholds and are compatible with integrated management of sorghum aphids. While most tests of these biological alternatives have shown that they have aphidicidal potential against sorghum aphids, it is crucial to take into account that their effectiveness in the field depends on a number of abiotic and biotic factors, including soil texture, temperature, humidity, and natural enemies.

Xanthotoxin: An Aphicidal Coumarin From Ficus petiolaris against Melanaphis sacchari Zehntner (Hemiptera: Aphididae).

In this research, we evaluated the aphicidal effect of the ethanolic extract of stems and bark of Ficus petiolaris Kunth (Moraceae), in laboratory bioassays in an artificial diet against apterous adult females of Melanaphis sacchari Zehntner (Hemiptera: Aphididae). The extract was evaluated at different concentrations (500, 1,000, 1,500, 2,000, and 2,500 ppm), and the highest percentage of mortality (82%) was found at 2,500 ppm after 72 h. The positive control imidacloprid (Confial®) at 1% eliminated 100% of the aphids, and the negative control (artificial diet) only presented mortality of 4%. The chemical fractionation of the stem and bark extract of F. petiolaris yielded five fractions of FpR1-5, which were each evaluated at 250, 500, 750, and 1,000 ppm. FpR2 had the strongest aphicidal effect, with 89% mortality at 72 h at 1,000 ppm. The pure xanthotoxin compound extracted from this fraction was even more effective, with 91% aphid mortality after 72 h at 100 ppm. The lethal concentration (LC50) of xanthotoxin was 58.7 ppm (72 h). Our results indicate that the extract of F. petiolaris showed toxic activity against this aphid, and its xanthotoxin compound showed strong aphicidal activity at low concentrations.

Percepción de riesgos en la salud de la comunidad indígena Mixteca El Calvario, al sur de México / Perception of health risks in the Mixteca El Calvario indigenous community in southern Mexico

La situación de salud y percepción de riesgos de las comunidades indígenas se mide por el grado sociodemográfico y estudio etnográfico. Nuestra población en estudio “El Calvario” es la única comunidad indígena Mixteca de Chilpancingo, Guerrero, conformada por 21 familias que se dedican a la agricultura, producción de maguey y mezcal, y elaboración de algunas artesanías de palma. El objetivo fue estudiar las condiciones de Calidad de Vida y percepción de riesgos en dos grupos vulnerables adultos y niños. En el caso de los adultos, se hizo por encuestas y en el caso de los niños se usó el dibujo. En los habitantes se perciben en riesgos de salud y de perder su identidad indígena, debido a que solo hablan su lengua materna los adultos, no saben leer y ni escribir. Así mismo, es inminente el riesgo de sufrir daños materiales debido a que las viviendas no son adecuadas. La migración de jóvenes, la perdida de cultura, el padecer enfermedades metabólicas debido a la dieta alta en refrescos. Así mismo, los niños expresaron las deficiencias educativas. Finalmente, la población en general se encuentra olvidada por los organismos de gobierno actuales, en riesgo sanitario, y de perdida de cultura ya que los jóvenes migrarán a otros lugares a buscar nuevas oportunidades. Es un fuerte llamado de atención a la sociedad ya que, de no actuar en consecuencia con dicha problemática, se perdería la única comunidad Mixteca en Chilpancingo, Guerrero.(AU)

The health situation and risk perception of indigenous communities is measured by sociodemographic grade and ethnographic study. Our study population "El Calvario" is the only Mixtec indigenous community in Chilpancingo, Guerrero, made up of 21 families that are dedicated to agriculture, production of maguey and mezcal, and making some palm handicrafts. The objective was to study the conditions of Quality of Life and risk perception in two vulnerable groups, adults and children. In the case of adults, it was done through surveys and in the case of children, drawing was used. The inhabitants are perceived as being at risk of health and of losing their indigenous identity, because only adults speak their mother tongue, they do not know how to read and write. Likewise, the risk of suffering material damage is imminent due to the inadequate housing. The migration of young people, the loss of culture, suffering from metabolic diseases due to a diet high in soft drinks. Likewise, the children expressed educational deficiencies. Finally, the general population is forgotten by current government agencies, at risk of health, and loss of culture as young people will migrate to other places to seek new opportunities. It is a strong call for attention to society since, if it does not act accordingly, the only Mixtec community in Chilpancingo, Guerrero, would be lost.(AU)