Glucanases and Chitinases in Mangifera indica: Identification, Classification, Phylogeny, and Expression Analysis of Defense Genes against Colletotrichum spp.

Plant glucanases and chitinases are defense proteins that participate in pathogenesis; however, very little is known about the glucanase (GLUC) and chitinase (CHIT) gene families in mango. Some mango cultivars are of great economic importance and can be affected by anthracnose, a postharvest disease caused by fungi of the genus Colletotrichum spp. This study identified and characterized 23 putative glucanases and 16 chitinases in the mango genome cv. Tommy Atkins. We used phylogenetic analyses to classify the glucanases into three subclasses (A, B, and C) and the chitinases into four classes (I, II, IV, and V). Information on the salicylic, jasmonic acid, and ethylene pathways was obtained by analyzing the cis-elements of the GLUC and CHIT class I and IV gene promoters. The expression profile of GLUC, CHIT class I, and CHIT class IV genes in mango cv. Ataulfo inoculated with two Colletotrichum spp. revealed different profile expression related to these fungi's level of virulence. In general, this study provides the basis for the functional validation of these target genes with which the regulatory mechanisms used by glucanases and chitinases as defense proteins in mango can be elucidated.

Ischemic gangrene of the penis due to Fournier's gangrene following traumatic transurethral catheterization: A case report.

Fournier's gangrene is a life-threatening fascial infection. We report a case of a 65-year-old diabetic man with a history of obstructive prostate growth who developed ischemic penile gangrene following traumatic transurethral catheterization. Despite initial debridement, progressive necrosis necessitated total penectomy and cystostomy. This case highlights the potential for devastating consequences of failed catheterization and the importance of early urologic intervention.

The Need and Opportunity to Update the Inventory of Plant Pathogenic Fungi and Oomycetes in Mexico.

Mexico generates specific phytosanitary regulations for each product and origin to prevent the entry of quarantine pests and/or delay their spread within the national territory, including fungi and oomycetes. Phytosanitary regulations are established based on available information on the presence or absence of these pathogens in the country; however, the compilation and precise analysis of reports is a challenging task due to many publications lacking scientific rigor in determining the presence of a taxon of phytosanitary interest in the country. This review evaluated various studies reporting the presence of plant pathogenic fungi and oomycetes in Mexico and concluded that some lists of diseases and phytopathogenic organisms lack technical-scientific basis. Thus, it highlights the need and presents an excellent opportunity to establish a National Collection of Fungal Cultures and a National Herbarium for obligate parasites, as well as to generate a National Database of Phytopathogenic Fungi and Oomycetes present in Mexico, supported by the combination of morphological, molecular, epidemiological, pathogenicity, symptom, and micrograph data. If realized, this would have a direct impact on many future applications related to various topics, including quarantines, risk analysis, biodiversity studies, and monitoring of fungicide resistance, among others.

Mechanisms of cell size regulation in slow-growing Escherichia coli cells: discriminating models beyond the adder.

Under ideal conditions, Escherichia coli cells divide after adding a fixed cell size, a strategy known as the adder. This concept applies to various microbes and is often explained as the division that occurs after a certain number of stages, associated with the accumulation of precursor proteins at a rate proportional to cell size. However, under poor media conditions, E. coli cells exhibit a different size regulation. They are smaller and follow a sizer-like division strategy where the added size is inversely proportional to the size at birth. We explore three potential causes for this deviation: degradation of the precursor protein and two models where the propensity for accumulation depends on the cell size: a nonlinear accumulation rate, and accumulation starting at a threshold size termed the commitment size. These models fit the mean trends but predict different distributions given the birth size. To quantify the precision of the models to explain the data, we used the Akaike information criterion and compared them to open datasets of slow-growing E. coli cells in different media. We found that none of the models alone can consistently explain the data. However, the degradation model better explains the division strategy when cells are larger, whereas size-related models (power-law and commitment size) account for smaller cells. Our methodology proposes a data-based method in which different mechanisms can be tested systematically.

First Report of Golovinomyces bolayi Causing Powdery Mildew on Lettuce in Mexico.

In August 2022, powdery mildew symptoms were detected on lettuce (Lactuca sativa) in a commercial field located in Quecholac, Puebla, Mexico. Signs appeared as whitish powdery masses on leaves. Disease incidence was about 100% and signs covered up to 40% of leaf surface. Mycelium was amphigenous forming white patches. Hyphal appressoria were indistinct or nipple-shaped and solitary. Conidiophores (n= 30) were hyaline, erect, arising from the upper surface of hyphal mother cells or lateral, and of 90 to 201 µm long. Foot cells were cylindrical, of 49 to 92 × 10-15 µm, followed by 1-3 shorter cells, and forming conidia in chains. Conidia (n= 100) were hyaline, ellipsoid-ovoid, doliiform-subcylindrical, 27 to 40 × 14 to 20 µm. Conidial germination belonging to the Euoidium type. Chasmothecia were not observed. The morphological characters were consistent with those of Golovinomyces bolayi (Braun et al. 2019). A voucher specimen was deposited in the Herbarium of the Department of Agricultural Parasitology at the Chapingo Autonomous University under accession number UACH451. To confirm the identification of the fungus, genomic DNA was extracted from conidia and mycelium following the CTAB method (Doyle and Doyle 1990), and the internal transcribed spacer (ITS) region was amplified by PCR using the primers ITS5/ITS4 (White et al. 1990) and sequenced. The resulting 506 bp sequence had 100% identity to those of G. bolayi (LC417109 and LC417106). Phylogenetic analyses using the Maximum Likelihood and Maximum Parsimony methods were performed and confirmed the results obtained in the morphological analysis. The isolate UACH451 grouped in a clade with isolates of G. bolayi. The ITS sequence was deposited in GenBank under accession number OR467546. Pathogenicity was confirmed by gently dusting conidia onto ten leaves of healthy lettuce plants. Five non-inoculated leaves served as controls. The plants were maintained in a greenhouse at 25 to 30 ºC, and relative humidity of 70%. All inoculated leaves developed similar symptoms to the original observation after 10 days, whereas control leaves remained disease free. Microscopic examination of the fungus on inoculated leaves showed that it was morphologically identical to that originally observed. Based on morphological data and phylogenetic analysis, the fungus was identified as G. bolayi. This pathogen has been previously reported causing powdery mildew on lettuce in Argentina, Canada, Chile, Ecuador, Peru, USA and Venezuela (Braun et al. 2019; Mieslerová et al. 2020). To our knowledge, this is the first report of G. bolayi causing powdery mildew on lettuce in Mexico.

Antimicrobial susceptibility of mastitis pathogens isolated from North American dairy cattle, 2011-2022.

A total of 10,890 bacterial isolates of Streptococcus dysgalactiae, Streptococcus uberis, Staphylococcus aureus and Escherichia coli isolated as etiological agents from dairy cows with mastitis by 29 veterinary laboratories across North America between 2011 and 2022 were tested for in vitro antimicrobial susceptibility by broth microdilution to ampicillin, cefoperazone, ceftiofur, cephalothin, erythromycin, oxacillin, penicillin-novobiocin and pirlimycin according to CLSI standards. Using available clinical breakpoints, antimicrobial resistance among S. dysgalactiae (n = 2406) was low for penicillin-novobiocin (0% resistance), ceftiofur (0.1%), erythromycin (3.2%) and pirlimycin (4.6%). Among S. uberis (n = 2398), resistance was low for ampicillin (0%) and ceftiofur (0.2%) and moderate for erythromycin (11.9%) and pirlimycin (18.4%). For S. aureus (n = 3194), resistance was low for penicillin-novobiocin (0%), ceftiofur (0.1%), oxacillin (0.2%), erythromycin (0.7%), cefoperazone (1.2%) and pirlimycin (2.8%). For E. coli (n = 2892), resistance was low for ceftiofur (2.8%) and cefoperazone (3.4%) and moderate for ampicillin (9.2%). Overall, the results indicate that mastitis pathogens in the United States and Canada have not shown any substantial changes in the in vitro susceptibility to antimicrobial drugs over the 12 years of the study, or among that of the proceeding survey from 2002-2010. The data support the conclusion that resistance to common antimicrobial drugs among mastitis pathogens, even to drugs that have been used in dairies for mastitis management for many years, continues to remain low.

Neurobiological basis of reinforcement-based decision making in adults with ADHD treated with lisdexamfetamine dimesylate: Preliminary findings and implications for mechanisms influencing clinical improvement.

BACKGROUND: ADHD is often described as a disorder of altered reward sensitivity, yet few studies have examined the extent to which: (i) treatments for ADHD impact reward-related mechanisms; and (ii) changes in the reward system are associated with clinical improvement. This study addresses these issues - examining the extent to which clinical improvement following lisdexamfetamine (LDX) treatment is associated with changes in brain reward system activation. METHODS: Twenty adults (M = 11, 55%, F = 9, 45%), ages 19-52 (M = 33.9, SD = 10.0) with ADHD participated in a randomized cross-over study with lisdexamfetamine (LDX) and placebo (PB). Changes in brain activation were assessed during functional magnetic resonance (fMRI) scans: after receiving 3-5 weeks of treatment with LDX and 3-5 weeks of no drug/PB. fMRI contrasts were derived from the passive-avoidance (PA) learning task, which assessed reward-related learning using computational variables. We analyzed the following conditions: the Choice-Phase, modulated by the expected value (EV; i.e., object-choose and object-reject), and the Feedback-Phase, modulated by the prediction error (PE; i.e., reward and punish). Clinical symptom severity was assessed via interview with the ADHD-Rating Scale (ADHD-RS-IV). To address the primary objective, we performed group-level mass-univariate regression analyses between LDX and PB of percent change of the ADHD-RS total scores and the four contrast images under the Choice- and Feedback-conditions. Significance was set at a whole-brain voxel-wise threshold of p < 0.05 with family-wise error (FWE) correction and an extent (cluster) threshold of 50 contiguous voxels. RESULTS: Improvement in ADHD symptoms with LDX was accompanied by significantly increased activation in a series of brain regions previously implicated in reinforcement processing in the choice and feedback conditions (e.g., left caudate and putamen, right orbitofrontal cortex, left middle frontal, superior frontal, and precentral gyri). CONCLUSIONS: These findings, while preliminary, are the first to show that ADHD symptom improvement with stimulant treatment is associated with increased responsiveness of brain systems engaged in reward processing. Results support the hypothesis that LDX treatment may restore balance to dysfunction (e.g., hypoactivation) within the brain reward circuitry in adults with ADHD. Trial RegistrationClinicaltrials.gov Identifier: NCT01924429.

Viruses of plant-pathogenic fungi: a promising biocontrol strategy for Sclerotinia sclerotiorum.

Plant pathogenic fungi pose a significant and ongoing threat to agriculture and food security, causing economic losses and significantly reducing crop yields. Effectively managing these fungal diseases is crucial for sustaining agricultural productivity, and in this context, mycoviruses have emerged as a promising biocontrol option. These viruses alter the physiology of their fungal hosts and their interactions with the host plants. This review encompasses the extensive diversity of reported mycoviruses, including their taxonomic classification and range of fungal hosts. We highlight representative examples of mycoviruses that affect economically significant plant-pathogenic fungi and their distinctive characteristics, with a particular emphasis on mycoviruses impacting Sclerotinia sclerotiorum. These mycoviruses exhibit significant potential for biocontrol, supported by their specificity, efficacy, and environmental safety. This positions mycoviruses as valuable tools in crop protection against diseases caused by this pathogen, maintaining their study and application as promising research areas in agricultural biotechnology. The remarkable diversity of mycoviruses, coupled with their ability to infect a broad range of plant-pathogenic fungi, inspires optimism, and suggests that these viruses have the potential to serve as an effective management strategy against major fungi-causing plant diseases worldwide.

Association between clinical and surgical variables with postoperative outcomes in patients treated for intestinal obstruction for non-malignant conditions: a cross-sectional study.

Intestinal obstruction is considered a frequent surgical pathology related to previous surgical procedures. Many different factors can lead to different outcomes when surgical management is needed. Therefore, we aim to describe the factors related to morbidity and mortality in surgical management of IO in a single-center experience. Retrospective observational study with a prospective database, in which we described patients who underwent surgical management due to intestinal obstruction between 2004 and 2015. Demographics, perioperative data, surgical outcomes, morbidity, and mortality were described. 366 patients were included. Female were 54.6%. Mean age was 61.26. Laparoscopic approach was done in 21.8% and the conversion rate was 17.2%. Intestinal resection was performed in 37.9% of the cases. Postoperative complications were observed in 18.85%. Reintervention and mortality were 9.5% and 4.1% respectively. Laparoscopic approach shows lesser time of intestinal transit (mean 28.67 vs. mean 41.95 h), and restart of oral intake after surgery (mean 96.06 vs. mean 119.65) compared with open approach. Increased heart rate and intensive care unit length of stay were related with mortality (p 0.01 and 0.000 respectively). For morbidity, laparotomy and need and duration of ICU stay were related with any complication statistically significant (p 0.02, 0.008, 0.000 respectively). Patients with increased heart rate in the emergency room, decreased amount of intravenous fluids, need and higher length of stay in the intensive care unit, and delay in resuming oral intake after surgery appear to have poor outcomes. Laparoscopic approach seems to be a safe and feasible approach for intestinal obstruction in selected patients.

Mechanisms of Cell Size Regulation in Slow-Growing Escherichia coli Cells: Discriminating Models Beyond the Adder.

Under ideal conditions, Escherichia coli cells divide after adding a fixed cell size, a strategy known as the adder. This concept applies to various microbes and is often explained as the division that occurs after a certain number of stages, associated with the accumulation of precursor proteins at a rate proportional to cell size. However, under poor media conditions, E. coli cells exhibit a different size regulation. They are smaller and follow a sizer-like division strategy where the added size is inversely proportional to the size at birth. We explore three potential causes for this deviation: precursor protein degradation, nonlinear accumulation rate, and a threshold size termed the commitment size. These models fit mean trends but predict different distributions given the birth size. To validate these models, we used the Akaike information criterion and compared them to open datasets of slow-growing E. coli cells in different media. the degradation model could explain the division strategy for media where cells are larger, while the commitment size model could account for smaller cells. The power-law model, finally, better fits the data at intermediate regimes.

Giant squamous cell carcinoma developed on a mesh-related enterocutaneous fistula: A case report.

INTRODUCTION: Squamous cell carcinoma degeneration on enterocutaneous fistulas (EF) is infrequent. There are some reports of malignant conversion in Crohn's disease-associated fistulas. Literature about the malignant development of mesh-related EF is even more limited. PRESENTATION OF THE CASE: A 66-year-old patient who developed necrotizing pancreatitis was managed through an open necrosectomy approach with a prolonged open abdomen that derived an incisional hernia which was repaired using a synthetic mesh. Years later, the patient was admitted to the service because of hypovolemic shock due to gastrointestinal bleeding. An abdominal wound with mesh exposition and cloudy discharge was observed. A high-output enterocutaneous fistula diagnosis was established. After an institutional surgical committee, a surgical approach was defined, a 60 cm en-block resection of the involved small bowel was done, and the surgical specimen was obtained for histopathological analysis. DISCUSSION: The use of prosthetic mesh in the case of incisional hernias is associated with a higher incidence of complications. However, there is no evidence of the development of squamous cell carcinoma developed on a mesh-related enterocutaneous fistula. This is a condition associated with Chron's disease and its diagnosis should be suspected by the exacerbation of local signs and symptoms. The scarce literature published suggests that this pathology can be managed by radical surgery and even chemoradiation, the last one required only for patients with associated Chron's disease. CONCLUSION: Squamous Cell Carcinoma developed on a mesh-related enterocutaneous fistula is a rare condition with no classic signs and symptoms that allow diagnostic identification.

First Report of White Mold Caused by Sclerotinia sclerotiorum on Brussels Sprouts in Mexico.

The Brussels sprout (Brassica oleracea var. gemmifera) is a cruciferous vegetable with high health-promoting value and Mexico is one of the most valuable exporters worldwide (Data Mexico 2023). From September to November 2021, white mold symptoms (Rimmer et al. 2007) were observed in Brussels sprouts (cv. Confidant) fields in Tonatico, Estado de México, Mexico. Irregular, necrotic lesions were observed on leaves, whereas abundant white mycelium, and later black sclerotia were produced outside and inside of stems. Disease incidence ranged from 20 to 40% in five fields. For fungal isolation, symptomatic stem pieces were surface sterilized with 2% sodium hypochlorite for 2 min, rinsed in sterilized distilled water twice, placed on PDA medium, and incubated at 25°C in darkness for 3 days. Sclerotinia-like colonies were consistently obtained and six isolates were purified by the hyphal-tip method. Fungal colonies were white and fluffy. Irregular, black, and small (3 to 6 mm diameter) sclerotia were produced at the edge of colonies after 5 days of incubation. The morphological characters were consistent with those of Sclerotinia sclerotiorum (Saharan and Mehta 2008). Two representative isolates were selected for molecular analysis and pathogenicity tests. The isolates were deposited in the Culture Collection of Phytopathogenic Fungi at the Colegio Superior Agropecuario del Estado de Guerrero under the accession numbers CSAEG50 and CSAEG51. For molecular identification, genomic DNA was extracted, and the internal transcribed spacer (ITS) region was amplified by PCR and sequenced using the primer pair ITS5/ITS4 (White et al. 1990). The sequences were deposited in GenBank (accession nos. OQ878510 and OQ878511). BLASTn searches in GenBank showed 100% identity with the available sequences of Sclerotinia sclerotiorum (accession nos. OQ891471, OQ891472, HQ833448, and MT177216). A phylogenetic analysis using the Maximum Likelihood method placed isolates CSAEG50 and CSAEG51 in the same clade as S. sclerotiorum. Pathogenicity tests were performed by inoculating 10 healthy Brussels sprout seedlings (cv. Confidant) grown in pots. A mycelial plug was directly placed on the stem of each plant. Five uninoculated Brussels sprout seedlings were used as control. All plants were placed in a moist chamber at 25°C with a 12-h photoperiod for 2 days. White mold symptoms appeared on inoculated plants after 3 days, whereas control plants remained symptomless. The fungi were reisolated from the infected plants and found to be morphologically identical to the isolates used for inoculation, fulfilling Koch's postulates. Pathogenicity test was performed twice with similar results. Sclerotinia sclerotiorum has been previously reported to infect Brussels sprouts in the USA (Campbell 1947). To our knowledge, this is the first report of Sclerotinia sclerotiorum causing white mold of Brussels sprouts in Mexico. The disease is widely distributed in Brussels sprouts fields in the central region of Mexico, therefore additional studies are needed to develop effective disease-management strategies.

First Report of Peach Scab Caused by Cladosporium tenuissimum in Mexico.

Peach (Prunus persica) is one of the most popular stone fruits in the world. From 2019 to 2022, typical scab symptoms were observed on 70% of peach fruits in a commercial orchard in Tepeyahualco, Puebla, Mexico (19°30'38"N 97°30'57"W). Fruit symptoms are black circular lesions of 0.3 mm in diameter. The fungus was isolated from symptomatic fruit pieces that were surface sterilized with 1% sodium hypochlorite for 30 s, rinsed in autoclaved distilled water three times, placed on PDA medium, and incubated at 28°C in darkness for 9 days. Cladosporium-like colonies were isolated. Pure cultures were obtained by single spore culture. Colonies on PDA showed aerial mycelium abundant, smoke-grey, fluffy, and with margin glabrous to feathery. Conidiophores were solitary, long, intercalary conidia narrow erect, macro- and micronematous, straight or slightly flexuous, cylindrical-oblong, olivaceous-brown, and often subnodulose. Conidia (n= 50) catenate in branched chains, obovoid to limoniform, sometimes globose, aseptate, olivaceous-brown, apically rounded, 3.1 to 5.1 × 2.5 to 3.4 µm. Secondary ramoconidia (n= 50) were fusiform to cylindrical, smooth-walled, 0-1-septate, pale brown or pale olivaceous-brown, measuring 9.1 to 20.8 × 2.9 to 4.8 µm. Morphology was consistent to that described for Cladosporium tenuissimum (Bensch et al. 2012; 2018). A representative isolate was deposited in the Culture Collection of Phytopathogenic Fungi of the Department of Agricultural Parasitology at the Chapingo Autonomous University under the accession number UACH-Tepe2. To further confirm the morphological identification, total DNA was extracted using the cetyltrimethylammonium bromide method (Doyle and Doyle 1990). The internal transcribed spacer (ITS) region, partial sequences of the translation elongation factor 1-alpha (EF1-α) and actin (act) genes were amplified by PCR, and sequenced using the primer pairs ITS5/ITS4 (White et al. 1990), EF1-728F/986R, and ACT-512F/783R (Carbone and Kohn 1999), respectively. The sequences were deposited in GenBank under the accession numbers OL851529 (ITS), OM363733 (EF1-α), and OM363734 (act). BLASTn searches in GenBank showed 100% identity with available sequences of Cladosporium tenuissimum accession (ITS: MH810309; EF1-α: OL504967; act: MK314650). A phylogenetic analysis using the maximum likelihood method placed isolate UACH-Tepe2 in the same clade as C. tenuissimum. To verify the pathogenicity of the fungus, 20 healthy peach fruits were inoculated with four drops of 15 µl of a conidial suspension (1 × 106 spores /ml). Ten control fruit were treated with sterilized water. All the fruits were kept in a moist chamber at 25°C for 10 days. Circular and necrotic lesions were produced eight days after inoculation, whereas control fruits remained healthy. Pathogenicity test was conducted three times with similar results. Fungal colonies were reisolated from the artificially inoculated fruit, thus fulfilling Koch's postulates. Cladosporium tenuissimum has been previously reported to cause diseases on strawberry, cashew, papaya, and passionfruit in Brazil (Rosado et al. 2019; Santos et al. 2020), as well as diseases on pitaya, hydrangea, and carnation in China (Xu et al. 2020; Li et al. 2021; Xie et al. 2021). Cladosporium carpophilum is reported as the causal agent of peach scab. The environmental conditions for the development of C. carpophilum are 20-30 °C in warm humid areas (Lawrence and Zehr 1982), however, in this case the infection by C. tenuissinum occurred in a temperate semi-dry climate, with temperatures of 5 -15 °C and R.H. less than 50 % with an incidence of 80 %. To our knowledge, this is the first report of Cladosporium tenuissimum causing peach scab in Mexico and worldwide.

Higher-Dimensional Origin of Extended Black Hole Thermodynamics.

Holographic braneworlds are used to present a higher-dimensional origin of extended black hole thermodynamics. In this framework, classical, asymptotically anti-de Sitter black holes map to quantum black holes in one dimension less, with a conformal matter sector that backreacts on the brane geometry. Varying the brane tension alone leads to a dynamical cosmological constant on the brane, and, correspondingly, a variable pressure attributed to the brane black hole. Thus, standard thermodynamics in the bulk, including a work term coming from the brane, induces extended thermodynamics on the brane, exactly, to all orders in the backreaction. A microsopic interpretation of the extended thermodynamics of specific quantum black holes is given via double holography.

First Report of Bacterial Wilt of Eggplant (Solanum melongena) Caused by Ralstonia pseudosolanacearum in Mexico.

Bacterial wilt caused by the Ralstonia solanacearum species complex (RSSC) is a major disease of solanaceous crops worldwide. In May 2022, symptoms of wilting, yellowing, and reduced growth were observed on eggplant (Solanum melongena) cv. Barcelona in a commercial greenhouse located in Culiacán, Sinaloa, Mexico. The disease incidence was recorded up to 30%. Sections of stems from diseased plants showed discoloration of the vascular tissue and the pith. Colonies with typical RSSC morphology were isolated from five eggplant stems on Petri plates containing casamino acid-peptone-glucose (CPG) medium supplemented with 1% 2,3,5-triphenyltetrazolium chloride (TZC), and incubated at 25°C for 48-h (Schaad et al. 2001; Garcia et al. 2019). On CPG medium + TZC, white and irregular colonies with pinkish centers were observed. On King's B medium, mucoid and white colonies were produced. The strains were Gram-negative in the KOH test and were nonfluorescent on King's B medium. Strains were positive using commercial Rs ImmunoStrip® (Agdia, USA). For molecular identification, DNA was extracted, and the partial endoglucanase gene (egl) was amplified by PCR and sequenced using the primer pair Endo-F/Endo-R (Fegan and Prior 2005). BLASTn searches showed 100% identity with available sequences of R. pseudosolanacearum from Musa sp. in Colombia (MW016967) and from Eucalyptus pellita in Indonesia (MW748363, MW748376, MW748377, MW748379, MW748380, MW748382). To confirm the bacterial identity, DNA was amplified with the primers 759/760 (Opina et al. 1997) and Nmult21:1F/Nmult22:RR (Fegan and Prior 2005) to generate 280 and 144-bp amplicons for RSSC and phylotype I (= R. pseudosolanacearum), respectively. A phylogenetic analysis was performed using the Maximum Likelihood method and the strain was distinguished as R. pseudosolanacearum sequevar 14. The strain (CCLF369) is currently preserved in the Culture Collection of the Research Center for Food and Development (Culiacán, Sinaloa, Mexico) and the sequence was deposited in GenBank (accession number OQ559102). Pathogenicity tests were performed by injection of 20-µl of a bacterial suspension (108 CFU/ml) at the base of the stem of five eggplants cv. Barcelona. Five plants inoculated with sterile distilled water were used as control. Plants were kept in a greenhouse at 28/37°C (night/day) for 12 days. All inoculated plants exhibited wilting, chlorosis, and necrosis of leaves between 8 and 11 days after inoculation, whereas control plants remained asymptomatic. The bacterial strain was only isolated from symptomatic plants and confirmed to be R. pseudosolanacearum using the molecular techniques mentioned above, fulfilling Koch´s postulates. Ralstonia pseudosolanacearum has been previously reported to cause bacterial wilt of tomato in Sinaloa, Mexico (García-Estrada et al. 2023); however, to our knowledge, this is the first report of R. pseudosolanacearum infecting eggplant in Mexico. Further studies on epidemiology and management strategies for this disease are required on vegetable crops in Mexico.

Retrospective validation of parkland grading scale in a Latin-American high-volume center.

BACKGROUND: Increased complication rates following laparoscopic cholecystectomies have been described, likely related to surgical difficulty, anatomical variations, and gallbladder inflammation severity. Parkland Grading Scale (PGS) stratifies the severity of intraoperative findings to predict operative difficulty and complications. This study aims to validate PGS as a postoperative-outcome predictive tool, comparing its performance with Tokyo Guidelines Grading System (TGGS). METHODS: This is a single-center retrospective cohort study where PGS and TGGS performances were evaluated regarding intraoperative and postoperative outcomes. Both univariate and bivariate analyses were performed on each severity grading scale using STATA-SE 16.0 software. Additionally, we proposed a Logistic Regression Model for each scale. Their association with outcomes was compared between both scales by their Receiver Operating Characteristic Curve. RESULTS: 400 Patients were included. Grade 1 predominance was observed for both PGS and TGGS (47.36% and 25.3%, respectively). A positive association was observed between higher PGS grades and inpatient postoperative care, length of stay, ICU care, and antibiotic requirement. Based on the area under the ROC curve, better performance was observed for PGS over TGGS in the evaluated outcomes. CONCLUSION: PGS performed better than TGGS as a predictive tool for inpatient postoperative care, length of stay, ICU, and antibiotic requirement, especially in severe cases.

First Report of Colletotrichum tropicale Causing Anthracnose on Pitahaya Fruit in Mexico.

Pitahaya (Hylocereus spp.), also called dragon fruit, is a cultivated cactus that is native to Mexico as well as Central and South America. In October 2021, anthracnose symptoms were observed on fruit of pitahaya (Hylocereus costaricensis) in a commercial orchard located in Culiacán, Sinaloa, Mexico. Lesions on fruit were circular, sunken, dark brown and with halo. To fungal isolation, small pieces from adjacent tissue to lesions of symptomatic fruits were surface disinfested by immersion in a 2% sodium hypochlorite solution for 2 min, rinsed in sterile distilled water, and placed in Petri plates containing potato dextrose agar (PDA). The plates were incubated at 25 ºC for 5 days in darkness. Colletotrichum-like colonies were consistently observed on PDA and five monoconidial isolates were obtained. An isolate was selected as a representative for morphological identification, multilocus phylogenetic analysis, and pathogenicity tests. The isolate was deposited as CCLF186 in the Culture Collection of Phytopathogenic Fungi at the Research Center for Food and Development (Culiacán, Sinaloa). On PDA, initially white colonies turned grey with abundant orange conidia masses at 8 days after incubation at 25 ºC. Conidia were cylindrical, with ends rounded, aseptate, hyaline, and measuring 15.2 to 18.9 × 4.3 to 6.4 µm (n= 100). Appressoria were terminal, subglobose to clavate, of 7.4 to 11.6 × 5.9 to 8.2 µm (n= 30). Setae were not observed. These morphological characters were consistent with those reported for the Colletotrichum gloeosporioides species complex (Weir et al. 2012). To determine the phylogenetic identity of the isolate CCLF186, genomic DNA was extracted following the CTAB method (Doyle and Doyle 1990), and the internal transcribed spacer (ITS) region, the ApMat intergenic region, as well as partial sequences of actin (act) and glyceraldehyde-3-phosphate dehydrogenase (gapdh) genes were amplified and sequenced using the primers pairs ITS5/ITS4 (White et al. 1990), AM-F/AM-R (Silva et al. 2012), GDF/GDR, and ACT-512F/ACT-783R (Weir et al. 2012), respectively. The sequences were deposited in GenBank under accession nos. OP269659 (ITS), OP302778 (gapdh), OP302777 (act), and OP302779 (ApMat). BLASTn searches revealed high identity with sequences of C. tropicale (CBS 124949) for ITS (100%), ApMat (100%), act (100%), and gapdh (100%). A phylogenetic tree based on Bayesian inference and Maximum Likelihood methods, including published ITS, ApMat, act, and gapdh sequence datasets for isolates in the Colletotrichum gloeosporioides species complex was generated. The phylogenetic analysis based on the concatenated sequences clustered the isolate CCLF186 with the C. tropicale reference isolates. Pathogenicity of the isolate CCLF186 was confirmed on 10 healthy pitahaya fruits without wounds. A drop of a conidial suspension (1 × 105 spores/ml) was placed on two locations on each fruit. Ten control fruits were treated with sterilized water. The fruits were kept in a moist plastic chamber at 25°C and 12 h light/dark for 8 days. The pathogenicity test was repeated twice. All inoculated pitahaya fruits exhibited sunken and necrotic lesions 6 days after inoculation, whereas no symptoms were observed on the control fruits. The fungus was consistently re-isolated only from the diseased fruits and found to be morphologically identical to the isolate used for inoculation. Recently, C. tropicale causing anthracnose in dragon fruit (Selenicereus monacanthus) was reported from Philippines (Evallo et al. 2022). Now, this is the first report of C. tropicale causing fruit anthracnose in H. costaricensis in Mexico and worldwide. These findings provide a basis for research about the distribution and effective disease-management strategies.

Fungicide Sensitivity of Sclerotinia sclerotiorum from U.S. Soybean and Dry Bean, Compared to Different Regions and Climates.

Fungicide use is integral to reduce yield loss from Sclerotinia sclerotiorum on dry bean and soybean. Increasing fungicide use against this fungus may lead to resistance to the most common fungicides. Resistance has been reported in Brazil (Glycine max) and China (Brassica napus subsp. napus), however, few studies have investigated fungicide sensitivity of S. sclerotiorum in the United States. This work was conducted to determine if there was a difference in fungicide sensitivity of S. sclerotiorum isolates in the United States from: (i) dry bean versus soybean and (ii) fields with different frequencies of fungicide application. We further hypothesized that isolates with fungicide applications of a single active ingredient from tropical Brazil and subtropical Mexico were less sensitive than temperate U.S. isolates due to different management practices and climates. The EC50(D) fungicide sensitivity of 512 S. sclerotiorum isolates from the United States (443), Brazil (36), and Mexico (33) was determined using a discriminatory concentration (DC) previously identified for tetraconazole (2.0 ppm; EC50(D) range of 0.197 to 2.27 ppm), boscalid (0.2; 0.042 to 0.222), picoxystrobin (0.01; 0.006 to 0.027), and thiophanate-methyl, which had a qualitative DC of 10 ppm. Among the 10 least sensitive isolates to boscalid and picoxystrobin, 2 presented mutations known to confer resistance in the SdhB (qualitative) and SdhC (quantitative) genes; however, no strong resistance was found. This study established novel DCs that can be used for further resistance monitoring and baseline sensitivity of S. sclerotiorum to tetraconazole worldwide plus baseline sensitivity to boscalid in the United States.

First Report of Ralstonia pseudosolanacearum Causing Wilt Disease in Tomato (Solanum lycopersicum L.) Plants from Mexico.

Mexico produces more than four million tons of tomato fruits and ranks tenth worldwide. In February 2022, tomato plants in a greenhouse in Culiacan, Sinaloa State, were affected by wilt diseases with an incidence of 20% and irreversible wilt and death of the infected plants (severity up 70%). When cut stems from affected plants, a reddish to brown discoloration of the vascular system was observed and these were disinfected with 1% NaClO for 5 min and then placed in a humid chamber. Characteristic milky-white exudate was obtained. From that exudate, irregular, mucoid, and white colonies with pink centres were obtained on casamino peptone glucose (CPG) plates supplemented with 1% 2,3,5-triphenyl 15 tetrazolium chloride (TZC); these characteristics are typical of the Ralstonia solanacearum species complex (RSSC) (Garcia et al., 2019). Identification of the pathogen was done by PCR using specific primer pairs reported by Paudel et al. (2022), RssC-wF3 (5'-TATATATCCTCGACTTTTCCATGAAGCTGTG-3') - RssCwR3 (5'-CTATATATATACCCCACTTGTTGAGGAACTG-3') and Rpseu-wF5 (5'-TTTTATTTTTTTGGTGTCCGGGCCAAGATAG-3') - Rpseu-wR5 (5'- TTATATTACTCGAACGTGCTGCAAAACCACT-3'), which amplified fragments of 162 and 251 bp for RSSC and Ralstonia pseudosolanacearum, respectively. Additionally, 759 (5'-GTCGCCGTCAACTCACTTTCC-3') - 760 (5'-GTCGCCGTCAGCAATGCGGAATCG-3') (Opina, et al., 1997) and Nmult21:1F (5'-CGTTGATGAGGCGCGCAATTT-3') - Nmult22:RR (5'- TCGCTTGACCCTATAACGAGTA-3') (Fegan and Prior, 2005) were used to generate 282 and 144 bp amplicons for RSSC and phylotype I, respectively. Subsequen to making the specific detection, the representative strain ClnMx was used to generate a sequence for the endoglucanase (egl) gene for separation into sequevars by using the primers Endo-F (5'- ATGCATGCCGCTGGTCGCCGC-3') and Endo-R (5'-GCGTTGCCCGGCACGAACACC-3'), which amplified a fragment of 750 bp (Fegan et al., 1998). The egl sequence (GenBank Access ON542479) showed 100% identity with the well-defined R. pseudosolanacearum sequevar 14, which was isolated from tomato plants from Senegal (UW763, I-14 GenBank Access CP051174) (Steidl et al., 2021), as well as, the strain MAFF 301070 (GenBank Access AB508612) from Japanese tomato. For pathogenicity tests, four 1-month-old tomato plants were infected using an insulin syringe that contained a pure bacterial suspension with approximately 2x108 CFU/mL. For each plant, 20 µL was infiltrated into the axil of the third upper leaf, and for untreated controls, tomato plants were infiltrated with sterile water. All plants were kept at 28°C under greenhouse conditions. Symptoms resembling those observed in the field were observed in inoculated plants six days after inoculation, and the plant pathogen was recovered on TZC medium. To confirm the bacteria identification a PCR using the specific primer pairs mentioned early was carried out. In contrast, water-treated control plants remained healthy. Koch's postulates were carried out twice with similar results. Ralstonia solanacearum species complex (RSSC) causes severe economic losses in many countries of the world because of their capability to infect a wide range of host plants, including potato, tomato, eggplant, tobacco, and, banana, among others. Ralstonia pseudosolanacearum has been reported to cause tomato wilt disease mainly on the Afro-Eurasian continent in areas such as Senegal, Cambodia, and Japan (Klass et al., 2019). To our knowledge, this is the first report of R. pseudosolanacearum causing bacterial wilt diseases in tomato plants from Mexico and because, the control of this bacteria is a challenge by the long survival time in soil, water, and infected plant tissues, the identification of this important pathogen could provide relevant information for developing management strategies.