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A lytic bacteriophage, BAU.Micro_SLP-22, was isolated from drain water in search of bio-controlling agents against avian salmonellosis. The phage genome is comprised of 59,738 bp with 56.96% guanine-cytosine content, encoding 81 protein-coding genes containing no transfer RNAs, antibiotic resistance, virulence, temperate marker, and clustered regularly interspaced short palindromic repeat coding sequences.

The research aimed to establish a multidrug-resistant Klebsiella pneumoniae-induced genetic model for mastitis considering the alternative mechanisms of the DjlA-mediated CbpA protein regulation. The Whole Genome Sequencing of the newly isolated K. pneumoniae strain was conducted to annotate the frequently occurring antibiotic resistance and virulence factors following PCR and MALDI-TOF mass-spectrophotometry. Co-chaperon DjlA was identified and extracted via restriction digestion on PAGE. Based on the molecular string property analysis of different DnaJ and DnaK type genes, CbpA was identified to be regulated most by the DjlA protein during mastitis. Based on the quantum tunnel-cluster profiles, CbpA was modeled as a novel target for diversified biosynthetic, and chemosynthetic compounds. Pharmacokinetic and pharmacodynamic analyses were conducted to determine the maximal point-specificity of selective flavonoids in complexing with the CbpA macromolecule at molecular docking. The molecular dynamic simulation (100 ns) of each of the flavonoid-protein complexes was studied regarding the parameters RMSD, RMSF, Rg, SASA, MMGBSA, and intramolecular hydrogen bonds; where all of them resulted significantly. To ratify all the molecular dynamic simulation outputs, the potential stability of the flavonoids in complexing with CbpA can be remarked as Quercetin > Biochanin A > Kaempherol > Myricetin, which were all significant in comparison to the control Galangin. Finally, a comprehensive drug-gene interaction pathway for each of the flavonoids was developed to determine the simultaneous and quantitative-synergistic effects of different operons belonging to the DnaJ-type proteins on the metabolism of the tested pharmacophores in CbpA. Considering all the in vitro and in silico parameters, DjlA-mediated CbpA can be a novel target for the tested flavonoids as the potential therapeutics of mastitis as futuristic drugs., Competing Interests: Declarations. Competing interest: The authors have no conflict of interest at all with the others. Ethical approval: The ethical approval is authorized by AWEC, Bangladesh Agricultural University, Mymesningh regarding the project titled—“Polyvalent Vaccine Development to Prevent Mastitis in Dairy Cow” with the Grant ID: BAS-USDA LS-26/2020 (4th phase)., (© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Objectives: The emergence of antimicrobial-resistant and mastitis-associated Enterococcus faecalis and Enterococcus faecium is of great concern due to the huge economic losses associated with enterococcal infections. Here we report the draft genome sequences of E. faecalis and E. faecium strains that were isolated from raw milk samples obtained from mastitis-infected cows in Bangladesh., Methods: The two strains were isolated, identified, and genomic DNA was sequenced using the Illumina NextSeq 550 platform. The assembled contigs were analysed for virulence, antimicrobial resistance genes, and multilocus sequence type. The genomes were compared to previously reported E. faecalis and E. faecium genomes to generate core genome phylogenetic trees., Results: E. faecalis strain BR-MHR218Efa and E. faecium strain BR-MHR268Efe belonged to multilocus sequence types ST-190 and ST-22, respectively, both of which appear to represent relatively rare sequence types. BR-MHR268Efe harboured only one antibiotic resistance gene encoding resistance towards macrolides (lsa(A)), while BR-MHR218Efa harboured ten different antibiotic resistance genes encoding resistance to aminoglycosides (ant[6]-Ia, aph(3')-III), sulphonamides (aac(6')-II), lincosamides (lnu(B)), macrolides (erm(B)), MLSB antibiotics (msr(C)), tetracyclines (tet(M), tet(L)), trimethoprim (dfrG), and pleuromutilin-lincosamide-streptogramin A (lsa(E)). Virulence gene composition was different between the two isolates. BR-MHR218Efa harboured only two virulence genes involved in adherence (acm and scm). BR-MHR268Efe harboured eight complete virulence operons including three operons involved in adherence (Ace, Ebp pili, and EfaA), two operons involved in biofilm formation (BopD and Fsr), and three exoenzymes (gelatinase, hyaluronidase, SprE)., Conclusions: The genome sequences of the strains BR-MHR268Efe and BR-MHR218Efa will serve as a reference point for molecular epidemiological studies of mastitis-associated E. faecalis and E. faecium. Additionally, the findings will help understand the complex antimicrobial-resistance in livestock-assoiated Enterococci., Competing Interests: Competing interests None declared., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Objective: This study focused on the antibiogram profiling of Staphylococcus aureus and coagulase-negative Staphylococcus spp . (CoNS) and the detection of icaA and blaZ genes from bovine raw milk samples., Materials and Methods: Bovine milk samples were collected from dairy farms, and Staphylococcus spp. were isolated and identified via conventional and molecular screening. Disk diffusion test (DDT) was implemented to determine the resistance pattern. Biofilm and β-lactamase-producing Staphylococcus spp. were identified via amplification of the icaA and blaZ genes. Methicillin-resistant Staphylococcus aureus and CoNS were identified by DDT and PCR of the mecA gene., Results: From 63 samples, 35 were confirmed as Staphylococcus spp., of which 16 (25.39%) S. aureus isolates were coagulase-positive, while 19 (30.16%) were negative. PCR confirmed that 50% (8/16) of S. aureus and 36.84% (7/19) of CoNS possessed the icaA gene. All S. aureus isolates were found resistant to penicillin-G (P) both phenotypically and genotypically. The isolates were also resistant to erythromycin (ERY) and oxytetracycline (TET). While CoNS showed high to reduced resistance against P, TET, ERY, and azithromycin, no S. aureus isolates were resistant to sulfamethoxazole, while 10.53% of CoNS isolates were. All S. aureus and CoNS isolates were susceptible to vancomycin and gentamicin. MR was exhibited by 37.5% of S. aureus and 42.10% of CoNS isolates. Moreover, S. aureus and CoNS had 56.25% and 52.63% multidrug-resistant (MDR) isolates, respectively., Conclusion: The present study revealed the presence of a biofilm-producing, MDR staphylococcal strain in milk that might endanger consumers. Routine surveillance and monitoring, along with antimicrobial resistance learning, can reduce risks., Competing Interests: The authors declare no conflict of interest., (© The authors.)

Endogenous metabolism is primarily responsible for losses in sucrose content and processing quality in postharvest sugarbeet roots. The genes responsible for this metabolism and the transcriptional changes that regulate it, however, are largely unknown. To identify genes and metabolic pathways that participate in postharvest sugarbeet root metabolism and the transcriptional changes that contribute to their regulation, transcriptomic and metabolomic profiles were generated for sugarbeet roots at harvest and after 12, 40 and 120 d storage at 5 and 12°C and gene expression and metabolite concentration changes related to storage duration or temperature were identified. During storage, 8656 genes, or 34% of all expressed genes, and 225 metabolites, equivalent to 59% of detected metabolites, were altered in expression or concentration, indicating extensive transcriptional and metabolic changes in stored roots. These genes and metabolites contributed to a wide range of cellular and molecular functions, with carbohydrate metabolism being the function to which the greatest number of genes and metabolites classified. Because respiration has a central role in postharvest metabolism and is largely responsible for sucrose loss in sugarbeet roots, genes and metabolites involved in and correlated to respiration were identified. Seventy-five genes participating in respiration were differentially expressed during storage, including two bidirectional sugar transporter SWEET17 genes that highly correlated with respiration rate. Weighted gene co-expression network analysis identified 1896 additional genes that positively correlated with respiration rate and predicted a pyruvate kinase gene to be a central regulator or biomarker for respiration rate. Overall, these results reveal the extensive and diverse physiological and metabolic changes that occur in stored sugarbeet roots and identify genes with potential roles as regulators or biomarkers for respiratory sucrose loss., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Fugate, Eide, Lafta, Tehseen, Chu, Khan and Finger.)

The emergence of antimicrobial-resistant and mastitis-associated Staphylococcus aureus is of great concern due to the huge economic losses worldwide. Here, we report draft genome sequences of two Staphylococcus aureus strains which were isolated from raw milk samples obtained from mastitis-infected cows in Bangladesh. The strains were isolated and identified using conventional microbiological and molecular polymerase chain reaction (PCR) methods. Antibiotic susceptibility testing was performed. Genomic DNA of the two strains was extracted and the strains were sequenced using the Illumina NextSeq 550 platform. The assembled contigs were analyzed for virulence determinants, antimicrobial resistance genes, extra-chromosomal plasmids, and multi-locus sequence type (MLST). The genomes of the two strains were compared with other publicly available genome sequences of Staphylococcus aureus strains. The raw read sequences were downloaded and all sequence files were analyzed identically to generate core genome phylogenetic trees. The genome of BR-MHR281strain did not harbour any antibiotic resistance determinants, however BR-MHR220 strain harbored mecA and blaZ genes. Analysis of BR-MHR220 strain revealed that it was assigned to sequence type (ST-6), clonal complex (CC) 5 and spa type t304, while BR-MHR281 strain belonged to ST-2454, CC8, and harbored the spa type t7867. The findings of the present study and the genome sequences of BR-MHR220 and BR-MHR281 strains will provide data on the detection and genomic analysis and characterization of mastitis-associated Staphylococcus aureus in Bangladesh. In addition, the findings of the present study will serve as reference genomes for future molecular epidemiological studies and will provide significant data which help understand the prevalence, pathogenesis and antimicrobial resistance of mastitis-associated Staphylococcus aureus ., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Herein this study, we sequenced the genome of a multidrug-resistant Salmonella enterica serovar Typhimurium strain MBR-MFRK-23 isolated from the liver tissue of a diseased layer chicken. The 4,964,854-bp draft genome comprises 50 contigs with 50.5× coverage and 52.1% GC content and is typed as S. enterica sequence type 19., Competing Interests: The authors declare no conflict of interest.

Cercospora leaf spot (CLS) is the most destructive foliar disease in sugar beet ( Beta vulgaris ). It is caused by Cercospora beticola Sacc., a fungal pathogen that produces toxins and enzymes which affect membrane permeability and cause cell death during infection. In spite of its importance, little is known about the initial stages of leaf infection by C. beticola . Therefore, we investigated the progression of C. beticola on leaf tissues of susceptible and resistant sugar beet varieties at 12-h intervals during the first 5 days after inoculation using confocal microscopy. Inoculated leaf samples were collected and stored in DAB (3,3'-diaminobenzidine) solution until processed. Samples were stained with Alexa Fluor-488-WGA dye to visualize fungal structures. Fungal biomass accumulation, reactive oxygen species (ROS) production, and the area under the disease progress curve were evaluated and compared. ROS production was not detected on any variety before 36 h postinoculation (hpi). C. beticola biomass accumulation, percentage leaf cell death, and disease severity were all significantly greater in the susceptible variety compared with the resistant variety ( P < 0.05). Conidia penetrated directly through stomata between 48 to 60 hpi and produced appressoria on stomatal guard cells at 60 to 72 hpi in susceptible and resistant varieties, respectively. Penetration of hyphae inside the parenchymatous tissues varied in accordance with time postinoculation and varietal genotypes. Overall, this study provides a detailed account to date of events leading to CLS disease development in two contrasting varieties., Competing Interests: The author(s) declare no conflict of interest.

Cercospora leaf spot (CLS) is a destructive disease limiting sugar beet production and is managed using resistant cultivars, crop rotation, and timely applications of effective fungicides. Since 2016, its causal agent, Cercospora beticola , has been reported to be resistant to quinone outside inhibitors (QoIs) and to have reduced sensitive to demethylation inhibitors (DMIs) in sugar beet growing areas in North Dakota and Minnesota. Isolates of C. beticola resistant to QoIs, DMIs, and both QoIs and DMIs were collected from fields in Foxhome, Minnesota, in 2017. Fitness of these resistant isolates was compared with that of QoI- and DMI-sensitive isolates in laboratory and greenhouse studies. In the lab, mycelial growth, spore production, and spore germination were measured. The results showed that resistant isolates had significantly less mycelial growth and spore production than sensitive isolates, while no significant difference in spore germination was detected. In the greenhouse, six leaf-stage sugar beets were inoculated with a spore suspension made from each resistant group and incubated in separate humidity chambers. CLS disease severity was evaluated visually at 7, 14, and 21 days after inoculation (DAI), and the areas under disease progress curve (AUDPC) were calculated. Resistant isolates had significantly smaller AUDPC but still caused as high disease severity as the sensitive ones at 21 DAI. Although QoI- and/or DMI-resistant isolates had a relatively slower disease development, they still caused high disease severity and need to be factored in disease management practices., Competing Interests: The author(s) declare no conflict of interest.

Here, we report the draft genome sequences of two Escherichia coli strains that were isolated from raw milk samples obtained from lactating cows with mastitis in Bangladesh. One strain was assigned to a novel sequence type 13054, and the other strain belonged to sequence type 101., Competing Interests: The authors declare no conflict of interest.

Cercospora leaf spot (CLS; causal agent Cercospora beticola Sacc.) is endemic in many sugar beet production regions due to the widespread distribution of C. beticola and the inability of current management practices to provide complete control of the disease. Roots harvested from plants with CLS, therefore, are inevitably incorporated into sugar beet root storage piles, even though the effects of CLS on root storage properties are largely unknown. Research was conducted to determine the effects of CLS on storage properties including root respiration rate, sucrose loss, invert sugar accumulation, loss in recoverable sucrose yield, and changes in sucrose loss to molasses with respect to CLS disease severity and storage duration. Roots were obtained from plants with four levels of CLS severity in each of three production years, stored at 5°C and 95% relative humidity for up to 120 days, and evaluated for storage characteristics after 30, 90, and 120 days storage. No significant or repeatable effects of CLS on root respiration rate, sucrose loss, invert sugar accumulation, loss in recoverable sucrose yield, or change in sucrose loss to molasses were detected after 30, 90, or 120 days storage regardless of the severity of CLS disease symptoms. Therefore, no evidence was found that CLS accelerates sugar beet storage losses, and it is concluded that roots harvested from plants with CLS can be stored without additional or specialized precaution, regardless of CLS symptom severity., Competing Interests: The author(s) declare no conflict of interest.

Injuries sustained by sugarbeet ( Beta vulgaris L.) roots during harvest and postharvest operations seriously reduce the yield of white sugar produced from stored roots. Although wound healing is critically important to reduce losses, knowledge of these processes is limited for this crop as well as for roots in other species. To better understand the metabolic signals and changes that occur in wounded roots, dynamic changes in gene expression were determined by RNA sequencing and the activity of products from key genes identified in this analysis were determined in the 0.25 to 24 h following injury. Nearly five thousand differentially expressed genes that contribute to a wide range of cellular and molecular functions were identified in wounded roots. Highly upregulated genes included transcription factor genes, as well as genes involved in ethylene and jasmonic acid (JA) biosynthesis and signaling and phenolic compound biosynthesis and polymerization. Enzyme activities for key genes in ethylene and phenolic compound biosynthesis and polymerization also increased due to wounding. Results indicate that wounding causes a major reallocation of metabolism in sugarbeet taproots. Although both ethylene and JA are likely involved in triggering wound responses, the greater and more sustained upregulation of ethylene biosynthesis and signaling genes relative to those of JA, suggest a preeminence of ethylene signaling in wounded sugarbeet roots. Changes in gene expression and enzymes involved in phenolic compound metabolism additionally indicate that barriers synthesized to seal off wounds, such as suberin or lignin, are initiated within the first 24 h after injury., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Fugate, Finger, Lafta, Dogramaci and Khan.)

Background: Colibacillosis, caused by avian pathogenic Escherichia coli (APEC), is one of the most significant infectious diseases affecting poultry worldwide., Objectives: This study aimed to determine the genomic diversity, virulence factor genes (VFGs), and antimicrobial resistance genes (ARGs) in the APEC MTR&#95;BAU02 strain isolated from a layer chicken using whole-genome sequencing (WGS)., Methods: Paired-end (2 × 250) WGS was performed using Illumina MiSeq sequencer (Illumina, San Diego, CA) and de novo assembly was performed using SPAdes. Core genome multilocus sequence typing (cgMLST) analysis between APEC MTR&#95;BAU02 and all of the ST1196 E. coli strains retrieved from the National Center for Biotechnology Information (NCBI) GenBank database was performed using the BacWGSTdb 2.0 server. We utilized different databases to detect ARGs, VFGs, and genomic functional features of the APEC MTR&#95;BAU02 strain., Results: The complete genome of APEC MTR&#95;BAU02 consists of 94 contigs comprising 4,924,680 bp (51.1% guanine-cytosine [GC] content), including 4681 protein-coding sequences, one chromosome, and one plasmid, and was assigned to ST1196. The closest relatives of APEC MTR&#95;BAU02 were four isolates originating from human clinical specimens (diarrhetic stool) in Bangladesh and two clinical isolates originating from chicken in India, which differed by 694 core genome multilocus sequence typing (cgMLST) alleles. One hundred and twenty-two ARGs and 92 VFGs were identified in the APEC MTR&#95;BAU02 genome. Metabolic functional annotations detected 380 SEED subsystems including genes coding for carbohydrate metabolism, protein metabolism, cofactors, vitamins, prosthetic groups and pigments, respiration, membrane transport, stress response, motility and chemotaxis, and virulence, disease, and defense., Conclusion: This study reports the genome sequence of a multidrug-resistant APEC strain isolated from layer birds in Bangladesh. The ARGs and VFGs, widespread in APEC MTR&#95;BAU02, are similar to those found in human isolates, and highlight the growing threat of antimicrobial resistance in both poultry and humans., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background: Sugarbeet ( Beta vulgaris L.) roots are stored under conditions that cause roots to dehydrate, which increases postharvest losses. Although exogenous jasmonate applications can reduce drought stress in intact plants, their ability to alleviate the effects of dehydration in postharvest sugarbeet roots or other stored plant products is unknown. Research was conducted to determine whether jasmonate treatment could mitigate physiological responses to dehydration in postharvest sugarbeet roots., Methods: Freshly harvested sugarbeet roots were treated with 10 µM methyl jasmonate (MeJA) or water and stored under dehydrating and non-dehydrating storage conditions. Changes in fresh weight, respiration rate, wound healing, leaf regrowth, and proline metabolism of treated roots were investigated throughout eight weeks in storage., Results: Dehydrating storage conditions increased root weight loss, respiration rate, and proline accumulation and prevented leaf regrowth from the root crown. Under dehydrating conditions, MeJA treatment reduced root respiration rate, but only in severely dehydrated roots. MeJA treatment also hastened wound-healing, but only in the late stages of barrier formation. MeJA treatment did not impact root weight loss or proline accumulation under dehydrating conditions or leaf regrowth under non-dehydrating conditions. Both dehydration and MeJA treatment affected expression of genes involved in proline metabolism. In dehydrated roots, proline dehydrogenase expression declined 340-fold, suggesting that dehydration-induced proline accumulation was governed by reducing proline degradation. MeJA treatment altered proline biosynthetic and catabolic gene expression, with greatest effect in non-dehydrated roots. Overall, MeJA treatment alleviated physiological manifestations of dehydration stress in stored roots, although the beneficial effects were small. Postharvest jasmonate applications, therefore, are unlikely to significantly reduce dehydration-related storage losses in sugarbeet roots., Competing Interests: The authors declare there are no competing interests.

Bovine tuberculosis (bTb) is endemic in Bangladesh but the true prevalence has not yet been reported. Our objectives for this study were to determine the true prevalence and identify risk factors for bTb at the animal- and herd-level in Bangladesh. A total of 510 cows were randomly selected during January 2018 to December 2018. Caudal fold (CFT) and comparative cervical tuberculin tests (CCT) were serially interpreted. Animal- and herd-level risk factor data were collected using a pre-tested questionnaire. The hierarchical true prevalence of bTb was estimated within a Bayesian framework. The herd- and animal-level risk factors were identified using mixed effects logistic regression. The apparent prevalence of bTb was 20.6% [95% Confidence Interval (CI): 17.3; 24.3] based on CFT. The animal-level true prevalence of bTb was 21.9 (13.0; 32.4). The herd-level true prevalence in different regions varied from 41.9% to 88.8%. The region-level true prevalence was 49.9 (13.8; 91.2). There is a 100% certainty that herds from Bhaluka and Mymensingh Sadar upazilas are not free from bTb. The odds of bTb were 3.9 times (1.2; 12.6) higher in herds having more than four cows than those with ≤ 4 cows. On the other hand, the risk of bTb was 3.3 times higher (1.0; 10.5) in non-grazing cows than grazing cows. Crossbred cows were 2.9 times (1.5; 5.9) more likely to be infected with bTb than indigenous cows. The risk of bTb in animals with cough was 2.3 times (1.2; 4.3) higher than those without cough. Crossbred, non-grazing cows with cough should be targeted for bTb surveillance. Herds of the Mymensingh, Sadar and Bhaluka regions should be emphasized for bTb control programs. Estimation of Bayesian hierarchical true prevalence facilitates identification of areas with higher prevalence and can be used to indicate regions that where true prevalence exceeds a pre-specified critical threshold., Competing Interests: The authors have declared that no competing interests exist.

Avian pathogenic Escherichia coli (APEC) causes significant economic losses in poultry industries. Here, we determined for the first time in Bangladesh, the prevalence of APEC-associated virulence genes in E. coli isolated from layer farms and their antibiotic resistance patterns. A total of 99 samples comprising internal organs, feces, and air were collected from 32 layer farms. Isolation was performed by culturing samples on eosin-methylene blue agar plates, while the molecular detection of APEC was performed by PCR, and antibiograms were performed by disk diffusion. Among the samples, 36 were positive for the APEC-associated virulence genes fimC , iucD, and papC . Out of 36 isolates, 7, 18, and 11 were positive, respectively, for three virulence genes ( papC , fimC , and iucD ), two virulence genes, and a single virulence gene. Although the detection of virulence genes was significantly higher in the internal organs, the air and feces were also positive. The antibiograms revealed that all the isolates (100%) were resistant to ampicillin and tetracycline; 97.2%, to chloramphenicol and erythromycin; 55.5%, to enrofloxacin; 50.0%, to norfloxacin and ciprofloxacin; 19.4%, to streptomycin; 11.1%, to colistin; and 8.33%, to gentamicin. Interestingly, all the isolates were multidrug-resistant (MDR). Spearman's rank correlation coefficient analysis revealed the strongest significant correlation between norfloxacin and ciprofloxacin resistance. This is the first study in Bangladesh describing the molecular detection of APEC in layer farms. Isolated APEC can now be used for detailed genetic characterization and assessing the impact on public health.

Objective: This research work was conducted for the molecular characterization of the circulating foot-and-mouth disease (FMD) virus in Bangladesh and revealed out their serotype., Materials and Methods: The VP1 gene of six field isolates of FMD virus (FMDV) serotypes (two serotypes O, two serotypes A, and two serotypes Asia 1) was subjected for sequencing and phylogenetic analysis. Neighbor-joining trees were constructed by using the Molecular Evolutionary Genetics Analysis 6, having the field nucleotide sequences of FMDV and related sequences available in the GenBank., Results: The nucleotide sequences of the VP1 genes of serotypes O, A, and Asia-1 of the isolates revealed that overall isolates were 91%-100% similar to the isolates reported from Bangladesh and other neighboring countries. Among the isolates reported from Bangladesh, serotype O had 98%-100% identity, serotype A had 91%-100% identity, and serotype Asia-1 had 94%-100% identity. A phylogenetic analysis revealed that the FMDV serotype O PanAsia-02 sub-lineage was confirmed in Bangladesh under the Middle East-South Asian (ME-SA) topotype. On the other hand, we identified genotype VII (18) of Asia topotype (serotype A) and lineage C (serotype Asia-1)., Conclusion: The FMDV serotype O PanAsia-02 sub-lineage was confirmed in Bangladesh under the ME-SA topotype for the first time. The extensive cross-border animal movement from neighboring countries may act as the source of diversified FMDV serotypes in Bangladesh., Competing Interests: The author declares that they have no conflict of interests., (Copyright: © Journal of Advanced Veterinary and Animal Research.)

Minnesota and North Dakota combined contain 55% of the sugar beet production area in the United States, contributing to 49% of the nation's sugar beet production in 2018. Fusarium diseases caused by Fusarium oxysporum f. betae and F. secorum on sugar beet can cause significant reduction in both root yield and sucrose concentration and purity. The objective of this research was to identify an alternative artificial inoculation method to induce Fusarium diseases on sugar beet leaves and roots caused by both Fusarium spp. in greenhouse conditions to better aid in research efforts. We tested four inoculation methods, including barley to seed, barley to root, drenching, and cutting. and compared them with the conventional root-dipping inoculation method. The inoculation method of placing Fusarium -colonized barley seed close to sugar beet seed (barley to seed) caused levels of symptom severities on both leaves and roots similar to the root-dipping method. Because the traditional root-dipping method involves a laborious transplant process, use of infected barley seed as inoculum may serve as an alternative method in the evaluation of host resistance and pathogen virulence among Fusarium diseases by Fusarium spp. on sugar beet at the seed or seedling stage.

Background: Parkinson's disease (PD) is a progressive neurodegenerative disorder affecting a large number of elderly people worldwide. The current therapies for PD are symptom-based; they do not provide a cure but improve the quality of life. Muscular dysfunction is the hallmark clinical feature of PD and oxidative stress and inflammation play a critical role in its pathogenesis. Epalrestat is used for the treatment of diabetic neuropathy and is known to improve antioxidative defense mechanisms in the CNS. Therefore, in this study, we investigated the role of Epalrestat in the reserpine induced mouse model of PD., Method: We used Swiss Albino mice for the PD model and tested for akinesia/bradykinesia, muscular rigidity, palpebral ptosis, and tremor, as well as conducting swim and open field tests. Brain samples were used to determine oxidative stress parameters and infiltration of immune cells., Results: Epalrestat treatment significantly improved akinesia and bradykinesia, muscular dysfunctions, tremor level, and gait functions compared to the reserpine group. It also improved the latency in the swim test. Eplarestat significantly reduced lipid peroxidation and NO concentration in different brain tissues and increased the activity of antioxidative enzymes, glutathione, catalase, and superoxide dismutase. Furthermore, Epalrestat reduced neuroinflammation by reducing the number of infiltrating immune cells., Conclusion: Eplarestat improves muscular dysfunction in PD by reducing oxidative stress and inflammation., Competing Interests: None., (© 2019 The Authors. Animal Models and Experimental Medicine published by John Wiley & Sons Australia, Ltd on behalf of The Chinese Association for Laboratory Animal Sciences.)

Rhizoctonia crown and root rot of sugar beet ( Beta vulgaris L.), caused by Rhizoctonia solani , continues to be one of the important concerns for the beet industry in Minnesota and North Dakota. Use of resistant cultivars is an important strategy in the management of R. solani in combination with seed treatment and timely fungicide application during the growing season. The objective of this greenhouse study was to determine how sugar beet plants responded to increasing age in resistance to R. solani . Each of three seed companies provided three commercial cultivars with varying R. solani resistance levels: susceptible, moderately resistant, and resistant. Seed were planted at a weekly interval to create different plant age groups from seed to 10-week-old plants, with growing degree days (GDD) ranging from 0 to 1,519 thermal time (°Cd). Seed and plants were all simultaneously inoculated with R. solani AG2-2-infested barley grains. Twenty-eight days after inoculation, plants were pulled and washed, and roots were evaluated for disease severity. All cultivars were highly susceptible to R. solani when inoculated at seed to 3 weeks old (0 to 464°Cd). At 4 and 5 weeks of plant age (617 to 766°Cd), resistant cultivars started to show significant resistance to R. solani . Proportion of the affected roots with disease score ≥ 5 followed a sigmoid response, declining with increased GDD in moderately resistant and resistant cultivars, whereas it continued to decline linearly with increased GDD in susceptible cultivars. This study demonstrated that sugar beet cultivars, regardless of their assigned level of R. solani resistance, were highly susceptible to the pathogen before they reached the six- to eight-leaf stage at 4 to 5 weeks (617 to 766°Cd) after planting. Therefore, additional protection in the form of seed treatment or fungicide application may be required to protect sensitive sugar beet seed and seedlings in fields with a history of R. solani under favorable environmental conditions.

Beet necrotic yellow vein virus (BNYVV) is the causal agent of rhizomania, a disease of global importance to the sugar beet industry. The most widely implemented resistance gene to rhizomania to date is Rz1 , but resistance has been circumvented by resistance-breaking (RB) isolates worldwide. In an effort to gain greater understanding of the distribution of BNYVV and the nature of RB isolates in Minnesota and eastern North Dakota, sugar beet plants were grown in 594 soil samples obtained from production fields and subsequently were analyzed for the presence of BNYVV as well as coding variability in the viral P25 gene, the gene previously implicated in the RB pathotype. Baiting of virus from the soil with sugar beet varieties possessing no known resistance to rhizomania resulted in a disease incidence level of 10.6% in the region examined. Parallel baiting analysis of sugar beet genotypes possessing Rz1 , the more recently introgressed Rz2 , and with the combination of Rz1 + Rz2 resulted in a disease incidence level of 4.2, 1.0, and 0.8%, respectively. Virus sequences recovered from sugar beet bait plants possessing resistance genes Rz1 and/or Rz2 exhibited reduced genetic diversity in the P25 gene relative to those recovered from the susceptible genotype while confirming the hypervariable nature of the coding for amino acids (AAs) at position 67 and 68 in the P25 protein. In contrast to previous reports, we did not find an association between any one specific AA signature at these positions and the ability to circumvent Rz1 -mediated resistance. The data document ongoing virulence development in BNYVV populations to previously resistant varieties and provide a baseline for the analysis of genetic change in the virus population that may accompany the implementation of new resistance genes to manage rhizomania.

Rhizoctonia damping-off and crown and root rot caused by Rhizoctonia solani are major diseases of sugar beet (Beta vulgaris L.) worldwide, and growers in the United States rely on fungicides for disease management. Sensitivity of R. solani to fungicides was evaluated in vitro using a mycelial radial growth assay and by evaluating disease severity on R. solani AG 2-2 inoculated plants treated with fungicides in the greenhouse. The mean concentration that caused 50% mycelial growth inhibition (EC 50 ) values for baseline isolates (collected before the fungicides were registered for sugar beet) were 49.7, 97.1, 0.3, 0.2, and 0.9 μg ml -1 and for nonbaseline isolates (collected after registration and use of fungicides) were 296.1, 341.7, 0.9, 0.2, and 0.6 μg ml -1 for azoxystrobin, trifloxystrobin, pyraclostrobin, penthiopyrad, and prothioconazole, respectively. The mean EC 50 values of azoxystrobin, trifloxystrobin, and pyraclostrobin significantly increased in the nonbaseline isolates compared with baseline isolates, with a resistant factor of 6.0, 3.5, and 3.0, respectively. Frequency of isolates with EC 50 values >10 μg ml -1 for azoxystrobin and trifloxystrobin increased from 25% in baseline isolates to 80% in nonbaseline isolates. Although sensitivity of nonbaseline isolates of R. solani to quinone outside inhibitors decreased, these fungicides at labeled rates were still effective at controlling the pathogen under greenhouse conditions.

Cercospora leaf spot (CLS) of sugar beet is caused by the fungus Cercospora beticola. CLS management practices include the application of the sterol demethylation inhibitor (DMI) fungicides tetraconazole, difenoconazole, and prothioconazole. Evaluating resistance to DMIs is a major focus for CLS fungicide resistance management. Isolates were collected in 1997 and 1998 (baseline sensitivity to tetraconazole, prothioconazole, or difenoconazole) and 2007 through 2010 from the major sugar-beet-growing regions of Minnesota and North Dakota and assessed for in vitro sensitivity to two or three DMI fungicides. Most (47%) isolates collected in 1997-98 exhibited 50% effective concentration (EC 50 ) values for tetraconazole of <0.01 μg ml -1 , whereas no isolates could be found in this EC 50 range in 2010. Since 2007, annual median and mean tetraconazole EC 50 values have generally been increasing, and the frequency of isolates with EC 50 values >0.11 μg ml -1 increased from 2008 to 2010. In contrast, the frequency of isolates with EC 50 values for prothioconazole of >1.0 μg ml -1 has been decreasing since 2007. Annual median difenoconazole EC 50 values appears to be stable, although annual mean EC 50 values generally have been increasing for this fungicide. Although EC 50 values are important for gauging fungicide sensitivity trends, a rigorous comparison of the relationship between in vitro EC 50 values and loss of fungicide efficacy in planta has not been conducted for C. beticola. To explore this, 12 isolates exhibiting a wide range of tetraconazole EC 50 values were inoculated to sugar beet but no tetraconazole was applied. No relationship was found between isolate EC 50 value and disease severity. To assess whether EC 50 values are related to fungicide efficacy in planta, sugar beet plants were sprayed with various dilutions of Eminent, the commercial formulation of tetraconazole, and subsequently inoculated with isolates that exhibited very low, medium, or high tetraconazole EC 50 values. The high EC 50 isolate caused significantly more disease than isolates with medium or very low EC 50 values at the field application rate and most reduced rates. Because in vitro sensitivity testing is typically carried out with the active ingredient of the commercial fungicide, we investigated whether loss of disease control was the same for tetraconazole as for the commercial product Eminent. The high EC 50 isolate caused more disease on plants treated with tetraconazole than Eminent but disease severity was not different between plants inoculated with the very low EC 50 isolate.

In all, 98 isolates of three Fusarium spp. (18 Fusarium oxysporum, 30 F. graminearum, and 50 Fusarium sp. nov.) obtained from sugar beet in Minnesota were characterized for pathogenicity and virulence on sugar beet in the greenhouse by a bare-root inoculation method. Among the 98 isolates tested, 80% of isolates were pathogenic: 83% of the F. oxysporum isolates, 57% of the F. graminearum isolates, and 92% of the Fusarium sp. nov. isolates. Symptoms varied from slight to moderate wilting of the foliage, interveinal chlorosis and necrosis, and vascular discoloration of the taproot without any external root symptoms. Among the pathogenic isolates, 14% were highly virulent and 12% were moderately virulent. Most of the highly virulent isolates (91%) and moderately virulent isolates (89%) were Fusarium sp. nov. All pathogenic isolates of F. graminearum and most pathogenic isolates (87%) of F. oxysporum were less virulent. In general, more-virulent isolates induced first foliar symptoms earlier compared with less-virulent isolates. This study indicates that both F. oxysporum and Fusarium sp. nov. should be used in greenhouse and be present in field studies used for screening and developing sugar beet cultivars resistant to Fusarium yellows complex for Minnesota and North Dakota.

Cercospora leaf spot, caused by the fungus Cercospora beticola Sacc., is the most serious and important foliar disease of sugar beet (Beta vulgaris L.) wherever it is grown worldwide. Cercospora leaf spot first caused economic damage in North Dakota and Minnesota in 1980, and the disease is now endemic. This is the largest production area for sugar beet in the United States, producing 5.5 to 6.0 million metric tons on approximately 300,000 ha, which is 56% of the sugar beet production in the United States. This Plant Disease feature article details a cooperative effort among the participants in the sugar beet industry in this growing area and represents a successful collaboration and team effort to confront and change a fungicide resistance crisis to a fungicide success program. As a case study of success for managing fungicide resistance, it will serve as an example to other pathogen-fungicide systems and provide inspiration and ideas for long-term disease management by fungicides.

Cercospora beticola survives as stromata in infected crop residue. Spores produced on these survival structures serve as primary inoculum during the next cropping season. This study was conducted to determine how long C. beticola can survive at different soil depths, the mechanism of inoculum dispersal, and the primary infection site in sugar beet. Longevity of C. beticola was studied over a 3-year period under field conditions at Fargo, ND. C. beticola-infected leaves were placed at depths of 0, 10, and 20 cm and retrieved after 10, 22, and 34 months. Survival of C. beticola inoculum declined with time and soil depth. Inoculum left on the soil surface, 0 cm in depth, survived the longest (22 months) compared with that buried at 10 cm (10 months) and 20 cm (10 months). C. beticola dispersal from the primary source of inoculum was studied in the field for three growing seasons. Sugar beet plants were surrounded with plastic cages with and without ground cover, or exposed with and without ground cover. Significantly higher disease severity was observed on exposed plants than caged plants with or without ground cover, suggesting that wind was the major dispersal factor for C. beticola inoculum. The primary infection site by C. beticola was determined in a greenhouse study. Leaves, roots, and stems of healthy sugar beet plants were inoculated with C. beticola. Cercospora leaf spot symptoms were observed only on plants that were leaf inoculated, suggesting that the leaf was the primary infection site for C. beticola.

Management of Cercospora leaf spot, caused by Cercospora beticola, is necessary for the economic production of sugar beet (Beta vulgaris). The objectives of this study were to evaluate the impact of two relative humidity thresholds (87 and 90%) on the daily infection values (DIVs) used to determine when fungicide applications were required, to determine whether current Cercospora management recommendations for northern areas of Minnesota and North Dakota could be used by growers in the southern areas of these states, and to compare the utility of calendar-based fungicide applications with the Cercospora management model. Research was conducted in Breckenridge, MN and St. Thomas, ND in 2003 and 2004. Fungicide applications significantly (P = 0.05) reduced maximum disease severity (y max ) and area under the disease progress curve (AUDPC) when compared with the nontreated control at both locations during 2003 and 2004. Fungicides applied according to DIVs calculated at RH ≥ 87% or RH > 90% gave similar results. The mandatory second fungicide application 14 days after the first application for southern areas did not significantly decrease disease severity or AUDPC, or improve root yield or recoverable sucrose compared with treatments without the mandatory application. This research illustrates that a DIV calculated at RH ≥ 87% would result in similar timing of fungicide applications compared with DIVs calculated at RH > 90%. The results further show that the recommendation of fungicide applications at initial symptom and subsequent applications based on DIV and disease severity should be used for both northern and southern growers. Finally, this research showed that fungicide applications based on the Cercospora management model provided similar, effective disease control with fewer fungicide applications compared with calendar-based applications.

Powdery mildew caused by Erysiphe polygoni was widespread on sugar beet (Beta vulgaris) in North Dakota during 2006. This disease is generally not prevalent in the state because of the application of fungicides, which also have efficacy against powdery mildew, for control of Cercospora leaf spot caused by Cercospora beticola. Because Cercospora leaf spot pressure was low in 2006, fewer fungicide applications were made in the state, thus allowing for more observations of powdery mildew. Leaf samples from four fields near Amenia, Minto, Prosper, and St. Thomas, ND were collected in mid-September to look for the perfect stage of E. polygoni, since this has recently been observed in Idaho, Colorado, Montana, and Nebraska (1-3). Only the leaves collected from the field near Amenia had visible immature (yellow and brown) globose ascomata; ascomata were not observed on the leaves collected in the other fields. Additional leaves were collected from the field near Amenia in early October; these leaves had immature and mature (black) globose ascomata that were 70 to 105 μm in diameter. Mature ascomata contained ovoid to elliptic asci with one to four hyaline-to-golden pigmented ascospores (20 to 25 × 12 to 20 μm). The color, shape, and size of ascomata, asci, and ascospores were similar to previously reported observations (1-4). The prevalence of the perfect stage in North Dakota is unknown, since no statewide surveys were conducted. To our knowledge, this is the first report of the perfect stage of E. polygoni on sugar beet in North Dakota. The occurrence of the perfect stage could lead to a means for overwintering in this area. Because of the means for genetic recombination, the risk of fungicide resistance and the development of races may increase. References: (1) J. J. Gallian and L. E. Hanson. Plant Dis. 87:200, 2003. (2) R. M. Harveson. Plant Dis. 88:1049, 2004. (3) B. Jacobsen et al. Plant Dis. 89:1362, 2005. (4) E. G. Ruppel. Powdery mildew. Pages 13-15 in: Compendium of Beet Diseases and Insects. E. D. Whitney and J. E. Duffus, eds. The American Phytopathological Society. St. Paul, MN, 1986.

In 2002, somel sugar beet (Beta vulgaris L.) fields in the Red River Valley (RRV) of Minnesota and North Dakota had symptoms characteristic of Fusarium yellows (4). In 2004, ≈5% of fields in the RRV had symptomatic plants. Interveinal yellowing of older leaves typically began in mid-July and as the disease progressed, younger leaves turned yellow. Sometimes, one side of the leaf was yellow or necrotic while the other side remained green. As leaves died, they remained attached to the crown. Transverse sections of roots revealed a light gray-brown discoloration of the vascular tissue but no external rotting of roots. Isolations from 35 symptomatic roots collected in eight fields yielded 25 isolates identified as F. oxysporum (from single conidia grown on homemade potato dextrose agar and carnation leaf agar) (3). Pathogenicity was determined by dipping roots of 5-week-old sugar beet plants (cv. ACH 9363) in a suspension of 10 4 conidia per ml for 8 min (12 isolates, 10 to 12 plants per isolate). Plants were planted in Cone-tainers (3.8 cm diameter × 21 cm; Stuewe and Sons, Inc. Corvallis, OR) containing sterile soil. Three known cultures of F. oxysporum Schlecht. emend. Snyd. & Hans. f. sp. betae Stewart (= F. conglutinans var. betae Stewart [4]) also were included (13 and 216c from L. Hanson, USDA-ARS, Fort Collins, CO; 0-1122 from The Pennyslvania State University Fusarium Research Center). The control was sterile water. Plants were placed in a greenhouse at 24 to 27°C with natural light supplemented with illumination from high-pressure sodium-vapor lamps for 16 h daily and lightly fertilized biweekly to avoid chlorosis from nutrient deficiency. After 6 to 7 weeks, plants were rated for disease on a 0 to 4 scale: 0 = no disease; 1 = slight to extreme plant stunting, leaves may be wilted; 2 = chlorotic leaves, some with necrosis at margins; 3 = tap root dried and brown to black in color, leaves dying; and 4 = plant dead (1). The experiment was repeated. Disease severity differed between trials, but all isolates of F. oxysporum and F. oxysporum f. sp. betae resulted in disease ratings statistically (P < 0.05) greater than that of the water control. In Trial 1, isolates of F. oxysporum averaged a rating of 2.1 (range of 1.8 to 3.3) and F. oxysporum f. sp. betae averaged 2.1 (range of 2.0 to 2.2) compared with 0.1 for the water control. One isolate of F. oxysporum had a statistically higher rating than did the cultures of F. oxysporum f. sp. betae. In Trial 2, isolates of F. oxysporum averaged a rating of 3.3 (range of 2.7 to 3.7) and F. oxysporum f. sp. betae averaged 3.1 (range of 2.7 to 3.4) compared with 0.2 for the water control. Cultures of F. oxysporum (8 of 12) resulted in ratings statistically higher than that of the least pathogenic culture of F. oxysporum f. sp. betae. Cultures of F. oxysporum and F. oxysporum f. sp. betae recovered from inoculated plants were identical to those used to inoculate plants. To our knowledge, this is the first report of F. oxysporum f. sp. betae on sugar beet in the Red River Valley of Minnesota and North Dakota. The disease has been reported in California, Colorado, Montana, Nebraska, Oregon, Texas, and Wyoming (1,2). References: (1) R. A. Cramer et al. J. Phytopathol. 151:352, 2003. (2) G. A. Fisher and J. S. Gerik. Phytopathology 84:1098, 1994. (3) P. E. Nelson et al. Fusarium Species: An illustrated Manual for Identification. The Pennsylvania State University Press. University Park, 1983. (4) D. Stewart. Phytopathology 21:59, 1931.