Infection Levels Research Articles

Seeds Priming with Bio-Silver Nanoparticles Protects Pea (Pisum sativum L.) Seedlings Against Selected Fungal Pathogens.

Nano-priming is a relatively new seed treatment technique using metal and metal oxide nanoparticles (NPs), and such application of NPs may support the plants' immunity. Recently we have shown that the that biologically synthesized silver nanoparticles (bio-AgNPs) used as short-term foliar treatment protect pea seedlings against D. pinodes and F. avenaceum. In the present study, the protection of peas against both fungal pathogens via seed priming with bio-AgNPs was analyzed. Moreover, the changes in the polar metabolic profiles of the seedlings caused by priming and infection were also compared. Seed priming with bio-AgNPs at concentrations of 50 and 100 mg/L considerably reduced the symptoms and infection levels of both pathogens by over 70% and 90% for F. avenaceum and D. pinodes, respectively. Pathogens infection and nano-priming affected the metabolic profile of pea seedlings. The major changes in the primary metabolism were observed among carbohydrates and amino acids. In turn, this may result in changes in the expression and accumulation of secondary metabolites. Therefore, further investigation of the effect of nano-priming should focus on the changes in the secondary metabolism.

A single immunization of Borreliella burgdorferi-infected mice with Vanguard crLyme elicits robust antibody responses to diverse strains and variants of outer surface protein C.

Lyme disease, caused by Borreliella burgdorferi and related species, is a growing health threat to companion animals across North America and Europe. Vaccination is an important preventive tool used widely in dogs living in, or near, endemic regions. In this report, we assessed anti-outer surface protein (Osp) A and anti-OspC antibody responses in B. burgdorferi-infected and -naïve mice (C3H/HeN) after immunization with a murine-optimized single dose of the Lyme disease subunit vaccine, Vanguard crLyme. crLyme is comprised of OspA and an OspC chimeritope-based immunogen designated as CH14. Mice that were infected and immunized developed higher levels of anti-OspC antibodies (Abs) than those infected only or that received one vaccine dose. The anti-OspC Abs that developed in the infected/immunized mice bound to all OspC variants tested (n = 22), whereas OspC Abs in serum from infected mice bound predominantly to the OspC variant (type A) produced by the infecting B. burgdorferi strain. Consistent with the absence of OspA expression in infected mammals, none of the infected mice developed Abs to OspA and did not develop anti-OspA Abs after single dose immunization. Lastly, serum from infected/immunized mice displayed significantly higher and broader killing activity than serum from non-immunized infected mice. The results of this study demonstrate that a single vaccination of actively infected mice results in strong anti-OspC Ab responses. This study contributes to our understanding of Ab responses to vaccination in actively infected mammals.

Dual-lock-and-key virus-mimicking nanoprobes for ultra-high accurate and sensitive imaging of viral infections in vivo

Fluorescence in situ imaging of viral infection lesions in vivo is crucial for precise diagnosis of viral diseases and evaluation of the extent of viral infection. Nevertheless, achieving highly specific and sensitive fluorescence imaging of viral infection sites in vivo has posed a persistent challenge. Here, we developed a dual-lock-and-key virus-mimicking nanoprobe that consisted of polyamide dendrimers (PAMAM) loaded internally with molecular beacons double-triggered by apurinic/apyrimidinic nucleic acid endonuclease 1 (APE1) and viral RNA (vRNA), and surface-modified with the E protein of Japanese encephalitis virus (JEV). This activatable nanoprobe generated dramatically amplified fluorescent signals stimulated by expressed vRNA and APE1 during viral infection, enabling ultrahigh specific and sensitive imaging of the lesions of JEV infection in vivo. This study provides a potential approach for accurate and sensitive detection of viral infection levels and assessment of the efficacy of antiviral drugs in vivo.

Contribution to a study on the seroprevalence of Equine Infectious Anemia in Tiaret province northwestern Algeria

A serological survey was conducted in equines in the province of Tiaret, in order to estimate the prevalence level of infection of equine infectious anemia (EIA). This survey has allowed to study the rate of infection by EIAV equine, to clarify the current situation of the EIA in the province of Tiaret, constitute a first step towards a national approach to control the emergence of this disease. Serum samples were collected from 293 horses and donkeys 30 and tested by ELISA technique. All results obtained by the ELISA test proved totally negative for all the samples studied. The equine infectious anemia is a topical disease. It is not well known and rarely sought on the ground. Yet the consequences of an outbreak can be very heavy on the economy.

Demographic and genetic impacts of powdery mildew in a young oak (Quercus robur L.) cohort

Key messageBy monitoring a field experiment over nine years, we investigated the impacts of the two main pathogen species Erysiphe quercicola S. Takam. and U. Braun and Erysiphe alphitoides (Griffon and Maubl.) U. Braun and S. Takam causing powdery mildew on a young cohort of pedunculate oak (Quercus robur L.), both from a demographic and genetic point of view using SNP markers. We show that survival rate is affected by mean disease severity. But while the growth-related tolerance to infection of the oak individual seems to be more determinant than resistance against infection, no equalizing effect of the disease could be detected.ContextStudies on the effects of pathogens on the survival and population dynamics of forest trees are scarce. Yet a better understanding of these interactions could prove strategic in the challenging context of climate change.AimsOur general objective was to characterize the demographic and genetic impact of the two main pathogen species Erysiphe quercicola S. Takam. and U. Braun and Erysiphe alphitoides (Griffon and Maubl.) U. Braun and S. Takam causing powdery mildew in the early stages of a Quercus robur L. population.MethodsAn ad hoc field design with two disease exposures, natural and protected, was surveyed over nine years. This enabled a detailed phenotypic monitoring of 1733 emerging individuals from 15 progenies, and the genotyping of 68% of them.ResultsThe pathogen induced high levels of seedling mortality several years after sowing, associated with reduced growth and capacity to overwinter. Fast-growing families showed the highest survival rate under both natural and protected disease exposure. Contrary to a possible trade-off hypothesis between growth and defense, family height potential was not negatively related to disease resistance across the studied oak mother trees. While supporting a deleterious effect of very low individual heterozygosity on the probability of survival, average genomic diversity was not significantly affected by mortality associated with powdery mildew. Our study also points to a few candidate genes for several fitness-related traits.ConclusionOverall, our results suggest that in oak natural populations, infection levels (related to resistance sensu stricto) may be less determinant than growth-related tolerance to infection for the fate of seedlings. However, an equalizing effect of powdery mildew on relative oak genotype performances cannot be excluded at later stages.

Effects of environmental factors on host-parasite interaction patterns in backyard-tethered goats of Kerala, India

The environment is the most important stratum in the epidemiological triad of a parasitic disease and any variations in the environmental factors may decide the dynamic occurrence and existence of different lifecycle stages of these parasites. The present study investigated the correlations between key biometeorological and demographical parameters with the incidence of different gastrointestinal parasites and hemoparasites among goats. Four hundred and thirty-two goats were screened for parasitic infection in a yearlong survey conducted from July 2022 to June 2023 in the Teaching Veterinary Clinical Complex, Mannuthy, Kerala, India. The weather parameters (Tmax, Tmin, RH, THI, and bright sunshine hours), air quality parameters [AQI, PM2.5, and PM10], and demographic parameters (gender and age) were recorded along with the test positivity of different categories of gastrointestinal parasites and hemoparasites in goats by routine fecal sample examination and blood smear examination, respectively. The infection level was ranked based on the severity of the infection. The mean and daily variations in biometeorological parameters were calculated and the data were statistically analyzed to figure out the pertinent correlations in host-parasite-environment interaction patterns. High levels of parasitic infections with significant month-wise variations and climate-correlated peak infection patterns were noticed. The incidence of parasites was negatively correlated to all parameters except humidity, indicating more severe parasitism during monsoon months. The significant variations in the host-parasite interaction dynamics point towards the need for detailed explorations concerning the lifecycle of each specific parasite with a focus on the possible environment-favourable and resistant lifecycle stages. Future studies may be designed from a biometeorological perspective to develop a crucial understanding of host-parasite-environment interactions in goats ensuring sustainable goat farming.

Prevalence of Passalurus ambiguus in domestic rabbits (Oryctolagus cuniculus) in Karbala province, Iraq

Commercial rabbit farming is primarily focused on the production of rabbit meat, although the presence of gastrointestinal disorders can impede their productivity. Endoparasites significantly contribute to the development of several diseases in rabbits, leading to higher rates of illness and death. Passalurus ambiguus is a widely distributed nematode that commonly infects the intestines of rabbits and hares. Parasitic disease can have a substantial economic impact on the industry by reducing the commercial value of rabbit meat and skins, impeding growth and development, and leading to mortality. This study aims to examine the frequency of gastrointestinal parasites in domestic rabbits that are bred in Karbala province, Iraq. Throughout the trial, fecal samples were collected from a randomly selected group of 48 live rabbits. The feces are collected directly from the anus of each living animal. The samples were tested and the presence of eggs and nematodes was detected in the infected samples. Analysis of samples was carried out at the Parasitology Laboratory, college of Veterinary Medicine, Kerbala university, using the flotation technique. Feces from 48 rabbits (ages 1-2) years. The feces are taken out of each animal directly from the anus. Upon testing, it was found that 16 out of 48 fecal samples, accounting for a prevalence of 33.33 %, were infected with at least one nematode. Chi-square (χ2) statistics. Age and genders are not statistically significant at P values ≥ 0.05. This study identified the presence of Passalurus ambiguus in rabbits of varying ages residing in Karbala region, Iraq, including both male and female domestic rabbits. Determining the age and gender of rabbits affected by passalurosis would enable the management of the infection level in these animals. Regular use of a consistent and dependable diagnostic method is essential in industrial rabbit farms to detect and monitor gastrointestinal parasite illnesses. Gastrointestinal parasites continue to be significant health issues that lead to the mortality of rabbit litters in production. Stress and a reduction in immune response may be crucial factors driving this phenomenon.

Molecular Characterization of Anaplasma spp. in Cattle from Kazakhstan.

Bovine anaplasmosis is an infectious vector-borne disease caused by bacteria of the genus Anaplasma, which have a wide global distribution and represent a high economic burden for agriculture. The use of molecular genetic techniques has increased our knowledge of the species diversity of Anaplasma spp. and naturally susceptible animals. Monitoring studies allow us to assess the level of infection in herds, as well as the involvement of natural vectors in the processes of maintaining and spreading infection. Despite the high prevalence of Theileria and Babesia in cattle in Kazakhstan, there is no information on the distribution and species diversity of Anaplasma spp in this country. As part of this work, 7027 DNA samples isolated from the whole blood of cattle from 175 settlements in all 17 Kazakhstan regions were PCR-tested for the presence of Anaplasma spp. Anaplasma carriers were found in 1.3% (90 out of 7027) of the tested animals in 9 of the 17 regions of Kazakhstan. The highest percentage of infected animals was recorded in Turkistan (South Kazakhstan) and North Kazakhstan with 4.46% and 2.48% positive samples, respectively. The partial sequencing of 16S rRNA and the groEL gene allowed us to identify five species of Anaplasma: A. centrale, A. marginale, Candidatus Anaplasma Mongolica, A. ovis, and Unknown Anaplasma with infection rates of 0.63%, 0.44%, 0.13%, 0.01%, and 0.01%, respectively.

Prevalence of Borrelia and Rickettsia in Ixodes ricinus from Chosen Urban and Protected Areas in Poland and the Czech Republic.

(1) Background: Ixodes ricinus is responsible for the spreading of medically important pathogens. Monitoring the level of tick infection in various areas is essential for determining the potential tick-born risk. This study aimed to detect Borrelia spp. and Rickettsia spp. in I. ricinus ticks collected in urban and protected areas both in Poland and the Czech Republic. (2) Methods: Ticks were collected by flagging in the years 2016-2017. Borrelia spp. was detected using nested PCR targeting the flaB gene and Rickettsia spp. using nested PCR targeting gltA. (3) Results: In total, DNA of Borrelia spp. was detected in 25.9% of samples. Ticks collected in Poland were more infected compared to the Czech Republic and ticks collected in protected areas were more infected with Borrelia spp. than ticks collected in urban areas. The RFLP analysis showed the occurrence of B. afzelii and B. garinii in both countries, and additionally B. valaisiana, B. burgdorferi s.s., and B. miyamotoi in Poland. Rickettsia spp. was detected in 17.4% of I. ricinus, with comparable infection level in both countries; however, regional differences were observed. (4) Conclusion: The regional differences in Borrelia spp. and Rickettsia spp. prevalence in I. ricinus indicate the complexity of factors influencing the level of infection and underline the need for adaptation public health surveillance strategies in each region.

Strength and durability of indirect protection against SARS-CoV-2 infection through vaccine and infection-acquired immunity.

Early investigation revealed that COVID-19 vaccines confer indirect protection to fully susceptible and unvaccinated persons, defined as a reduced risk of SARS-CoV-2 infection among social contacts of vaccinated individuals. However, indirect protection from infection-acquired immunity and its comparative strength and durability to vaccine-derived indirect protection in the current epidemiologic context of high levels of vaccination, prior infection, and novel variants are not well characterized. Here, we show that both infection-acquired and vaccine-derived immunity independently yield indirect protection to close social contacts with key differences in their strength and waning. Analyzing anonymized data from a system-wide SARS-CoV-2 surveillance program of 177,319 residents across 35 California state prisons from December 2021 to December 2022 in a case-control design, we find that vaccine-derived indirect protection against Omicron SARS-CoV-2 infection is strongest within three months post-vaccination [30% (95% confidence interval: 20-38%)] with subsequent modest protection, whereas infection-acquired immunity provides 38% (24-50%) indirect protection to roommates for 6 months after SARS-CoV-2 infection, with moderate indirect protection persisting for over one year. Variant-targeted vaccines (bivalent formulation including Omicron subvariants BA.4/BA.5) confer strong indirect protection for at least three months [40% (3-63%)]. These results have important implications for understanding the long-term transmission dynamics of SARS-CoV-2 and can guide vaccine policy and public health measures, especially in high-risk environments such as prisons.

Some Pro-inflammatory Cytokines Levels in Staphylococcal infection

Staphylococci is gram-positive, spherical cells, they are commonly distributed in the environment often part of normal human flora. Staph. aureus, Staph. saprophyticus, and Staph. epidermidis are the three most significant clinically important pathogens. Cytokines are small secreted proteins (<40 kDa), which are produced by nearly every cell to regulate and affect immune response (96). Proinflammatory cytokines are proteins that have been associated with cell proliferation, differentiation, and changes in gene expression, and synthesis of matrix proteins important to cell growth and tissue repair, these proteins are often released in a cascade, where an initial release of TNF is followed first by IL-1 and then by IL-6. 110 clinical samples include 110 throat swap and 60 saliva samples were collected from tonsillitis patients in Kirkuk of both gender and different age groups. Sterile cotton swab were used for throat swab and sterile container for saliva collection. The results showed that Staphylococcus aureus scored highest percentage (59%), followed by Staph. hemolyticus (15%) compared to Staph. epidermidis that scored lowest percentage (2.4%). A sensitivity test was carried out for all bacterial isolates, Staphylococcus spp. showed high resistance to Cefotoxin (, Benzyle-pencillin (, Ampicillin-Sulbactam(, Oxacillin (, Ceftriaxone (, and Erythromycin(, while Staphylococcus spp. showed high sensitive to Linezolid (100%), Teicoplanin (100%), Vancomycin (100%), and Rifampicin (100%). Staph. aureus scored high mean levels at IL-6 (14.51±13.1 ng/L) compared with other Staphylococcus spp. (0.52±0.1 ng/L), while Staph. aureus scored (90.47±77.79 pg/ml) compared with other Staphylococcus spp. (113.0±79.9 pg/ml).

Seasonal distribution and correlation between IL-10 and IL-13 gene polymorphism and their expression in scabies-infected patients.

Scabies is a significant concern in global health. It is more prevalent in individuals who have poor hygiene and live in crowded conditions, hence, it is seasonal distribution and immunological response in Erbil population is the aim of the present study. In the Erbil Dermatology Education Centre in Erbil, Iraq, 154 patients were recruited for the research between April 2022 and March 2023. If a patient has a suspicious skin lesion and itching for a minimum of one week, scabies may be considered. Blood samples were collected from each participant in the study to evaluate serum levels of IL-10 and IL-13, then the DNA was isolated to study the gene polymorphism for the mentioned cytokines. Results showed that female 60.3% were more infected than male 39.6%. The median age of participants was (10 - >51) years, among infested, adolescents aged 10-20 years displayed the highest rate (31.8%). The carriers of GA genotype of IL-10 were protective against the infection, OR:0.61. while the TT carriers of IL-13 were susceptible to scabies infection with OR:2.14. IL-10 GA genotype was more prevalent in male patients OR:2.14 whereas the AA genotype was most protective in females OR:0.32. the IL-13 CT genotype was protective for males with OR:0.52. Both of IL-10 and IL-13 serum levels were increased significantly with infection and highest levels were found in wild homozygous genotypes (GG and CC) and lowest ratio was found in mutant homozygous genotypes of IL-10 and IL-13 respectively. Point mutation in IL-10 GA was protective and wild TT genotype of IL-13 was susceptible to the scabies infection. Double mutation in IL-10 AA was protective to females and single mutation of IL-13 CT was protective to males.

Analysis of the species composition and infection of ticks with pathogens of Lyme disease in the territory of the Republic of Tatarstan for the period 2012–2022

BACKGROUND: The role of ixodid ticks in the transmission of pathogens of a number of natural focal infections can hardly be overestimated. Analysis of the manifestations of the epizootic process is important for planning preventive and anti-epidemic measures and is an integral component of epidemiological surveillance of natural focal infections. AIM: To analyze the species composition of ticks and the infection of ticks with borreliosis pathogens in the Republic of Tatarstan for the period 2012–2022. MATERIALS AND METHODS: An analysis of the species composition of ticks collected by the zoological group of the Center for Hygiene and Epidemiology in the Republic of Tatarstan from environmental objects during the period 2012–2022 was carried out. Testing of the ticks for Borrelia burgdorferi and Borrelia miyamotoi was carried out using the polymerase chain reaction method with hybridization-fluorescent detection. An analysis of statistical observation form No. 2 “Information on infectious and parasitic diseases” (the section “Tick bites”) for the Republic of Tatarstan from 2012 to 2022 was carried out. Statistical processing was performed using descriptive statistics methods in Microsoft Excel. RESULTS: The total number of people who applied for tick bites in the Republic of Tatarstan in 2012–2022 was 77,703, of which 59,168 (76.2%) were people over 18 years of age. During the study period, 5536 ticks were collected for research on the territory of the Republic of Tatarstan from environmental objects. The species composition of ticks was distributed as follows: Dermacentor reticulatus — 75.4%, Ixodes ricinus — 14.1%, Ixodes persulcatus — 10.5%. Infection of different types of ticks with Borrelia was not the same and was 20.4% for I. persulcatus, 14% for I. ricinus, 4.4% for D. reticulatus. In 4 regions of the republic, from ticks of the species I. ricinus pathogenic genomospecies B. miyamotoi were found (the infestation of this type of tick with B. miyamotoi was 0.4%). Indicators of morbidity of the population with ixodid tick-borne borreliosis and indicators of infection rates for ticks vary in different regions of the republic. CONCLUSIONS: The dominant species of ixodid ticks in natural epitopes of the Republic of Tatarstan in 2012–2022 were D. reticulatus (75.4% of the total number of collected ticks). For ticks of this species, the lowest level of infection with Borrelia was demonstrated (4.4% of ticks). This may be one of the factors responsible for the low, in comparison with other regions, incidence rates of tick-borreliosis in the Republic of Tatarstan. The identification of ticks of the species I. ricinus infected with B. miyamotoi confirms the need for targeted molecular genetic and serological examination for recurrent fever in patients with non-erythematous forms of febrile illness after an episode of tick bite.

Host cell glycosylation selects for infection with CCR5- versus CXCR4-tropic HIV-1.

Human immunodeficiency virus type 1 (HIV-1) infection involves a selection bottleneck that leads to transmission of one or a few variants. C-C motif chemokine receptor 5 (CCR5) or C-X-C motif chemokine receptor 4 (CXCR4) can act as coreceptors for HIV-1 viral entry. However, initial infection mostly occurs via CCR5, despite abundant expression of CXCR4 on target cells. The host factors that influence HIV-1 susceptibility and selection during transmission are unclear. Here we conduct CRISPR-Cas9 screens and identify SLC35A2 (a transporter of UDP-galactose expressed in target cells in blood and mucosa) as a potent and specific CXCR4-tropic restriction factor in primary target CD4+ T cells. SLC35A2 inactivation, which resulted in truncated glycans, not only increased CXCR4-tropic infection levels but also decreased those of CCR5-tropic strains consistently. Single-cycle infections demonstrated that the effect is cell-intrinsic. These data support a role for a host protein that influences glycan structure in regulating HIV-1 infection. Host cell glycosylation may, therefore, affect HIV-1 selection during transmission in vivo.

Exploring pathogen population density as a metric for understanding post-COVID infectious disease surges.

After the easing of COVID-19 restrictions, peaks of common infectious diseases surpassed pre-pandemic levels, raising questions about causes and ways to monitor these changes. A proposed measure, the Pathogen Population Density (PPD) score, could help track these shifts. PPD refers to the concentration of infectious agents within a population at a given time and location, serving as a potential indicator of infection levels in susceptible individuals at the population level. It is likely that PPD remains relatively stable within a specific community, as an equilibrium forms between infections and susceptibility. During the pandemic, nonpharmaceutical interventions (NPIs) led to a reduction in infectious diseases, possibly lowering population immunity and decreasing the PPD score. Once NPIs were lifted, the PPD score likely increased sharply due to a larger pool of susceptible individuals, causing more primary infections and stronger recurrent infections, faster transmission, and more severe pathogenic outcomes at the individual level. Monitoring the PPD score over time could help predict when infection peaks will occur. PPD is influenced by factors such as public health strategies, vaccination programs, and the behavior of high-risk individuals. As a quantitative measure, PPD has the potential to serve as a valuable predictive and monitoring tool, helping public health officials anticipate and track changes in infectious disease dynamics. It could be an effective tool for managing future outbreaks or pandemics and serve as a communication tool between scientists and the public to understand the emergence of new disease peaks.

Healthy lifestyle and HPV infection risk: results from a cross-sectional study.

Human Papillomavirus (HPV) infection constitutes a significant global public health challenge despite the widespread implementation of vaccination programs, with infection rates persistently high. Recent studies suggest that lifestyle factors including diet quality (DQ) and physical activity (PA) could play a pivotal role in the mitigation of HPV infections. This investigation explored the influence of DQ, PA, and a healthy lifestyle on the incidence of HPV infection in adult women. Data from 5,308 women aged 20-59 from the National Health and Nutrition Examination Survey (NHANES) 2007-2016. DQ and PA were measured using the Healthy Eating Index (HEI-2015) and the Global Physical Activity Questionnaire (GPAQ), respectively, creating four lifestyle groups based on how well they matched dietary and activity guidelines. The associations between key factors and HPV infection were explored using multivariate logistic regression, trend tests, and interaction tests. Fully adjusted multivariable logistic regression models revealed an inverse association between the risk of HPV infection and higher levels of PA (OR = 0.914; 95% CI: 0.854-0.979) as well as DQ (OR = 0.993; 95% CI: 0.989-0.998). Individuals in the highest tertile of DQ displayed a reduced risk of HPV infection relative to those in the lowest tertile (OR = 0.846; 95% CI: 0.726-0.986). Belonging to the third quintile of PA was linked to a lower risk of HPV infection than the lowest quintile (OR = 0.823; 95% CI: 0.681-0.995). Subgroups adhering to a healthy DQ were linked to a lower risk of HPV infection irrespective of PA meeting guideline recommendations. Our findings underscore the importance of a healthy diet in conjunction with appropriate PA in preventing HPV infection, offering new insights for public health policies and interventions.

Leveraging machine learning to discriminate wheat scab infection levels through hyperspectral reflectance and feature selection methods

Real-time or pre-symptomatic wheat scab (WS) detection is inevitable for precision agriculture to secure yield and quality at the critical grain formation stage. For this, feature selection (FS) techniques and machine learning (ML) have demonstrated their capabilities. However, for the same type and size of dataset, all FS and ML techniques behave differently due to their diverse primary constituents. This study attempts to leverage ML for WS classification and prediction employing different FS techniques on hyperspectral data of wheat spikes. The spectral features were selected and assessed to regress and classify disease occurrence. Relief-F-neural net (NN) manifested the best results with classification accuracy (CA) of 67 % and 89 % at the pre-symptomatic scale and 3 days after inoculation (DAI), respectively. Followed by continuous wavelet transform (CWT)-NN with 63 % CA at the pre-symptomatic scale and CWT-Xgboost with 89 % CA at 3DAI. For prediction, random forest regression revealed best accuracy of R2 = 0.94 and RMSE = 7.70, followed by partial least squares regression with R2 = 0.90 and RMSE = 10.37. The results offer a precise quantitative benchmark for future investigations into the capacity of hyperspectral data and FS for the real-time quantification of plant diseases.

A-273 Rapid, Portable Invasive Fungal Infection Diagnostic and Candida Antimicrobial Susceptibility Testing

Abstract Background Traumatic wounds are associated with a higher risk of invasive fungal infections (IFI), which lead to a significant increase in amputation or death. Actionable treatment decisions are hampered because true infections can be difficult to differentiate from external wound contaminates, and tissue biopsy and quantification of fungal load is required to determine if treatment is necessary, which currently takes several days. GeneCapture is developing the CAPTURE 60-minute, portable infection identification platform to detect infectious levels of common fungi including the genera Aspergillus, Fusarium, and Scedosporium, the family Herpotrichiellaceae, and the order Mucorales. The company is also pursuing rapid identification and antimicrobial susceptibility testing for Candida species from multiple infection types. Methods This study seeks to determine the Limit of Detection (LoD), sensitivity and specificity of the CAPTURE ID assay for fungal pathogens. Since tissue biopsy samples must be used to assess the presence of IFI in wounds, GeneCapture has developed a manual process for the crude homogenization of muscle tissue to allow this matrix to be used as the starting sample for the ID assay. Mock samples were tested for inclusivity and exclusivity on a series of IFI ID panels against multiple strains of the included fungi, against fungi not yet specifically detected on the panel, and against bacteria that commonly infect wounds. While the LoD for yeasts may be determined in CFU/mL, there is no similar, easy-to-define metric for molds. GeneCapture has developed a quantitative PCR method to detect the single copy beta-tubulin gene to roughly correlate fungal dry mass and nuclei number to assay signal for molds cultured in Potato Dextrose Broth. Determining beta-tubulin copy numbers for extracts from mock samples then allowed detection of the progression of IFI over the course of infection. Lastly, antimicrobial susceptibility testing (AST) was performed on two species of Candida to validate utilizing GeneCapture’s rapid phenotypic AST method for fungal infections. Results To date, the CAPTURE ID panel has been tested against 27 included or excluded strains of fungi or bacteria with 97% sensitivity and 100% specificity. The current LoD for mold organisms varies from approximately 1,000 nuclei for Aspergillus species, to 100,000 nuclei for Fusarium species. Additional work to understand this range is underway. Lastly, we have correctly determined the susceptibility of Candida species using our novel approach within 4-6 hours. Conclusions GeneCapture’s ID and AST technology offer a unique solution to address the increasing prevalence of fungal infections. Our platform can identify the most common fungal components in a wound within 60 minutes of receiving a tissue biopsy sample. Candida infections occur across multiple matrices and rapidly identifying the species of Candida and then determining its phenotypic susceptibility profile will have a large impact on treating these difficult pathogens.

B-351 Genotoxic Producing Escherichia coli Protects and Promotes Breast Adenocarcinoma

Abstract Background Recent studies have linked certain colibactin producing subphylas of E. coli to colorectal cancer. The bacterium synthesizes colibactin via the pks genomic island containing a series of clbA to clbS genes. The genotoxin causes inter-strand DNA cross-links, which lead to double strand breaks and promotion of colorectal adenocarcinoma. The published studies have prioritized the colorectal regions of the body where E. coli is commonly found, but none have identified the same genotoxic relationship with other types of adenocarcinoma. In this study, we evaluate DNA damage and cellular changes of mammary breast tissue based on high and low levels of colibactin producing E. coli exposure. The goal is to evaluate if colibactin can affect cells in the mammary breast tissue promoting breast adenocarcinoma growth. Methods Two cell lines were acquired from ATCC: primary mammary epithelial cells (PCS-600-010) and MCF-7 breast adenocarcinoma cells (HTB-22). Both cell lines were split into six culture flasks and maintained until ∼80% confluence was reached using the ATCC cell culture protocols. Once all the flasks had reached the target confluence, they were divided into exposure groups of high and low concentrations of colibactin producing E. coli, a group of non-colibactin producing E. coli, a group exposed only to phosphate buffered saline, and a control group with no exposure. Cellular morphology was observed using light microscopy before, after exposure, and every 5 minutes up to 15 minutes. E. coli and colibactin DNA concentrations were quantified using quantitative polymerase chain reaction. Genomic DNA was extracted from each group of cells. The DNA from each group was whole genome sequenced and analyzed for genomic variation. Results The normal cells exposed to E. coli showed immediate lysing and continued to lyse after a 5 minute incubation; however, the adenocarcinoma positive cells showed no lysing up to 30 minutes incubation. Sequencing results show genomic variation between the groups and specific break points associated with the E. coli exposed groups. Conclusions Our results show that the presence of E. coli with mammary breast tissue can destroy normal tissue, promote lysis resistance, and allow adenocarcinoma cells to continue to grow. The colibactin producing E. coli does cause double stranded breaks and can lead to cell death. This new information points out a critical need to monitor the microbiome and infection levels of people at risk or in early stages of cancer to prevent adenocarcinoma growth and destruction of normal tissue. This study indicates that colibactin producing E. coli may play a larger role in tissue damage and promotion of other types of cancer. More research is needed to elucidate the molecular pathway and mechanisms of colibactin, which will lead to new diagnostics or therapeutics that monitor E. coli levels or inhibit colibactin’s toxic effects.

Influence of Wheat Cultivars, Infection Level, and Climate after Anthesis on Efficacy of Fungicide for Control of Fusarium Head Blight in the Huang-Huai-Hai Plain of China

Fusarium head blight (FHB), caused by the Fusarium graminearum species complex, causes significant losses in grain yield and quality of wheat (Triticum aestivum) by inducing floret sterility. Grains become contaminated with mycotoxins, especially deoxynivalenol (DON), making them unsuitable for consumption. To clarify the impact of wheat cultivar resistance, infection level, and climate after anthesis on the efficacy of a fungicide for the control of FHB, we treated two moderately susceptible cultivars and 11 susceptible cultivars with fungicide (48% phenamacril + tebuconazole) at anthesis over two years. FHB incidence (INC), disease severity index (DSI), Fusarium-damaged kernels, DON contamination, thousand-kernel weight, and yield were evaluated under artificially inoculated and naturally infected field trials in 2018 and 2021. The results of multi-factor variance analysis show that the control efficacy with respect to INC and DSI is affected by cultivar, fungicide, infection level, and climatic conditions including the average daily temperature, average daily relative humidity, and total rainfall from anthesis to 21 days after anthesis (p < 0.01). Notably, cultivar resistance (deviance = 13.34, 9.55, and 11.22) is more important than fungicide (deviance = 5.77, 6.66, and 6.69) to control the efficacy of INC, DSI, and DON. The results also suggest that infection level appears to be more important than cultivars and fungicide to control the efficacy of INC, and more important than fungicide to control the efficacy of DSI. Total rainfall is more important than other climatic factors. Our results reveal that fungicide is more effective in moderately susceptible cultivars (‘Zhengmai 9023’ and ‘Xinong 979’, 89.5%~98.9%) and some susceptible cultivars than in other susceptible cultivars (‘Zhengmai 7698’ and ‘Zhoumai 27’, 51.9%~67.2%). Thus, integrating cultivar resistance with fungicide application can be an effective strategy for the management of FHB and DON in winter wheat in the Huang-huai-hai Plain of China.