Enhancing drought stress tolerance and growth promotion in chiltepin pepper (Capsicum annuum var. glabriusculum) through native Bacillus spp.

The drought can cause a decrease in food production and loss of biodiversity. In northern Mexico, an arid region, the chiltepin grows as a semi-domesticated crop that has been affected in its productivity and yield. An alternative to mitigate the effect of drought and aid in its conservation could be using Plant Growth-Promoting Bacteria (PGPB). The present study evaluated the capacity of native Bacillus spp., isolated from arid soils, as PGPBs and drought stress tolerance inducers in chiltepin under controlled conditions. Chiltepin seeds and seedlings were inoculated with native strains of Bacillus spp. isolated from arid soils, evaluating germination, vegetative, and drought stress tolerance parameters. The PGPBs improved vegetative parameters such as height, stem diameter, root length, and slenderness index in vitro. B. cereus (Bc25-7) improved in vitro survival of stressed seedlings by 68% at -1.02 MPa. Under greenhouse conditions, seedlings treated with PGPBs exhibited increases in root length (9.6%), stem diameter (13.68%), leaf fresh weight (69.87%), and chlorophyll content (38.15%). Bc25-7 alleviated severe water stress symptoms (7 days of water retention stress), and isolates B. thuringiensis (Bt24-4) and B. cereus (Bc25-7, and Bc30-2) increased Relative Water Content (RWC) by 51%. Additionally, the treated seeds showed improved germination parameters with a 46.42% increase in Germination Rate (GR). These findings suggest that using PGPBs could be an alternative to mitigate the effect of drought on chiltepin.

Pepper growth promotion and biocontrol against Xanthomonas euvesicatoria by Bacillus cereus and Bacillus thuringiensis formulations.

Background: Bacillus genus has been used in horticultural crops as a biocontrol agent against insect pests, microbial phytopathogens, and plant growth-promoting bacteria (PGPB), representing an alternative to agrochemicals. In particular, B. cereus (Bc) and B. thuringiensis (Bt) have been studied for their fungicidal and insecticidal activities. However, their use as biofertilizer formulations and biocontrol agents against phytopathogenic bacteria is limited. Objective: To evaluate Bc and Bt formulations as PGPB and biocontrol agents against the bacterial spot agent Xanthomonas euvesicatoria (Xe) in greenhouse-grown chili peppers. Methods: Bc and Bt isolates obtained from soil samples were identified and characterized using conventional biochemical and multiplex PCR identification methods. Bioassays to determine Bc and Bt isolates potential as PGPB were evaluated on chili pepper seedlings in seedbeds. In addition, formulations based on Bc (F-BC26 and F-BC08) and Bt (F-BT24) strains were assessed as biofertilizers on pepper, under controlled conditions. Furthermore, in vitro antagonism assays were performed by confronting Bc and Bt isolate formulations against Xe isolates in direct (foliage) and indirect (resistance induction) phytopathogen biocontrol assays on pepper plants, which were grown under controlled conditions for 15 d after formulations treatment. Results: Isolates were identified as Bc and Bt. Formulations significantly improved pepper growth in seedbeds and pots, whereas in vitro bioassays demonstrated the bactericidal effect of Bc and Bt strains against Xe isolates. Furthermore, assays showed significant plant protection by F-BC26, F-BC08, and F-BT24 formulated strains against Xe. Conclusion: Results indicated that F-BT24 and F-BC26 isolates formulations promoted pepper growth and protected it against Xanthomonas euvesicatoria.

Detection of rotavirus and norovirus among children with acute gastroenteritis in Merida and Chihuahua cities, Mexico.

INTRODUCTION: Infantile acute gastroenteritis (AGE) is a leading cause of morbidity and mortality, particularly in developing countries. The most frequent etiological agents of viral gastroenteritis in children are adenovirus, astrovirus, rotavirus, and norovirus, the last two, leading causes. Thus, the aim of this study was to identify the presence of these two viruses in children with AGE, from two cities located in the Southeast and the Northwest regions of México. METHODOLOGY: HuNoVs were detected and characterized by RT-PCR and sequencing, while RVs were detected by RNA electrophoresis. RESULTS: The presence of RV and HuNoV was evaluated in 81 stool samples; 37 were collected between April and July 2013 from patients with acute diarrhea in Merida, and 44 were collected between January and June 2017 in Chihuahua, who attended health services. Despite vaccination, RV resulted in the predominant viruses detected, with 30.8% (25/81) positivity, while HuNoV infection was present in 8.6% (7/81) of the stool samples; GII strains were identified circulating in the Southeast, while GI strains were identified in the Northwest. Moreover, co-infections with both viruses were detected at a prevalence rate of 2.4% (2/81). CONCLUSIONS: The circulation of RV and HuNoV in the country is continuous and should be constantly monitored due to their impact on public health.

Prevalence of Xanthomonas euvesicatoria (formally X. perforans) associated with bacterial spot severity in Capsicum annuum crops in South Central Chihuahua, Mexico.

BACKGROUND: Xanthomonas spp. causes bacterial spot disease, which reduces quality and yield of pepper crops in Mexico. Identification of phytopathogen species is necessary to implement more effective control strategies. OBJECTIVE: The aim of this study was to isolate and identify infecting Xanthomonas species in South Central Chihuahua pepper-producing areas. METHODS: Diseased plants were collected from 30 cultivation lots and bacteria were isolated from damaged tissues. Potential causative agents were isolated, identified, and characterized by biochemical and molecular analysis. Pathogenicity tests from each isolate were then performed on 30-d-old pepper seedlings, exposing five leaves to 10 µL of 1 × 108 CFU/mL bacterial suspensions of each isolate, using sterile distilled water as a control. Disease severity was determined after 10 d by calculating leaf damage percentage. Furthermore, we evaluated the susceptibility of the highest bacterial spot severity-causing isolates (13 isolates) to copper sulphate (CuS), copper gluconate (CuG), copper oxychloride + oxytetracycline hydrochloride (Cu + Ox), gentamicin + oxytetracycline hydrochloride (Gen + Ox), and gentamicin sulphate (GenS). Copper-resistance genes (copLAB) were detected by PCR analysis among isolates. RESULTS: Thirty-seven foliage isolates were identified as Xanthomonas euvesicatoria (14%), which were associated with bacterial spot disease in jalapeño pepper. Tested Xanthomonas isolates were resistant to Cu-based compounds, but susceptible to Cu + Ox. All isolates were susceptible to Gen + Ox and GenS. CopLAB genes were detected in all but one strain. CONCLUSIONS: X. euvesicatoria (formally X. perforans) may be considered as an emerging pathogen of bacterial spot pepper in Mexico. Among disease management strategies, alternatives to copper should be taken into consideration.

Streptophyta and Acetic Acid Bacteria Succession Promoted by Brass in Slow Sand Filter System Schmutzdeckes.

Macro- and microorganism activities are important for the effectiveness of the slow sand filtration (SSF), where native microorganisms remove contaminants mainly by substrate competition, predation, and antagonism. The aim of the present study was to evaluate the addition of the oligodynamic metals iron, copper, and brass, inserted separately into SSF to enhance pollutant removal in water samples. Four laboratory-scale SSFs were built and tested: control, iron, copper, and brass. Water analysis included physicochemical evaluation, total and fecal coliform quantification. An analysis on microbial communities in the SSFs schmutzdecke was achieved by using 16S rRNA amplification, the Illumina MiSeq platform, and the QIIME bioinformatics software. The results demonstrated that inorganic and organic contaminants such as coliforms were removed up to 90%. The addition of metals had no significant effect (p > 0.05) on the other parameters. The microbial community analysis demonstrated different compositions of the SSF with brass-influent, where the eukaryote Streptophyta was predominant (31.4%), followed by the acetic acid bacteria Gluconobacter (24.6%), and Acetobacteraceae (7.7%), these genera were absent in the other SSF treatments. In conclusion, the use of a SSF system can be a low cost alternative to reduce microbial contamination in water and thus reduce gastrointestinal diseases in rural areas.

Prevalence of Rotavirus Genogroup A and Norovirus Genogroup II in Bassaseachic Falls National Park Surface Waters in Chihuahua, Mexico.

In areas lacking potable water treatment, drinking contaminated water may represent a public health threat. In addition to enteropathogenic bacteria and parasites, fecal contamination in water environments is associated with the transmission of enteric viruses and other causal agents of infectious disease. Rotavirus and norovirus are the main enteric viral agents responsible for diarrheic outbreaks. The aim of the present study was to detect seasonal variation of rotavirus and norovirus in the surface water at Bassaseachic Falls National Park during 2013. Rivers and streams within and nearby this park were sampled once in each season during 2013. Viral concentration was carried out by a handmade filtration equipment, using a commercial electropositive membrane coupled with the virus absortion elution technique (VIRADEL©). Detection of rotavirus and norovirus was performed by SYBR Green reverse transcription-real time polymerase chain reaction (SYBR GREEN© RT-qPCR) analyses. Norovirus genogroup II was detected in samples collected in June and October 2013. In the case of rotavirus, genogroup A was detected in March and June. The presence of rotavirus and norovirus was related to viral acute diarrhea in children less than five years of age, who were inhabiting the sampled areas. This may indicates that the contaminated water was potentially a risk factor for regional diarrheic outbreaks.

Multidrug-Resistant Bacteria Isolated from Surface Water in Bassaseachic Falls National Park, Mexico.

Bacterial pathogens are a leading cause of waterborne disease, and may result in gastrointestinal outbreaks worldwide. Inhabitants of the Bassaseachic Falls National Park in Chihuahua, Mexico show seasonal gastroenteritis problems. This aim of this study was to detect enteropathogenic microorganisms responsible for diarrheal outbreaks in this area. In 2013, 49 surface water samples from 13 selected sampling sites along the Basaseachi waterfall and its main rivers, were collected during the spring, summer, autumn, and winter seasons. Fecal and total coliform counts were determined using standard methods; the AutoScan-4 system was used for identification of isolates and the antibiotic resistance profile by challenging each organism using 21 antibiotics. Significant differences among seasons were detected, where autumn samples resulted in the highest total (p < 0.05) and fecal (p < 0.001) coliform counts, whereas the lowest total coliform counts were recorded in spring. Significant differences between sampling sites were observed, where samples from sites 6, 8, and 11 had the highest total coliform counts (p < 0.009), whereas samples from site 9 exhibited the lowest one. From the microbiological analysis, 33 bacterial isolates from 13 different sites and four sampling seasons were selected; 53% of isolates were resistant to at least one antibiotic, and 15% exhibited a multidrug resistance (MDB) phenotype. MDB were identified as Klebsiella oxytoca (two out of four identified isolates), Escherichia coli (2/7), and Enterobacter cloacae (1/3). In addition, some water-borne microorganisms exhibited resistance to cefazoline, cefuroxime, ampicillin, and ampicillin-sulbactam. The presence of these microorganisms near rural settlements suggests that wastewater is the contamination source, providing one possible transmission mechanism for diarrheal outbreaks.