Microbiology Commons^™

Potent Bile Acid Microbial Metabolites Modulate Clostridium Perfringens Virulence, Tahrir Jatli Alenezi, Ying Fu, Bilal Alrubaye, Thamer Alanazi, Ayidh Almansour, Hong Wang, Xiaolun Sun

Poultry Science Faculty Publications and Presentations

Clostridium perfringens is a versatile pathogen, inducing diseases in the skin, intestine (such as chicken necrotic enteritis (NE)), and other organs. The classical sign of NE is the foul smell gas in the ballooned small intestine. We hypothesized that deoxycholic acid (DCA) reduced NE by inhibiting C. perfringens virulence signaling pathways. To evaluate the hypothesis, C. perfringens strains CP1 and wild-type (WT) HN13 and its mutants were cultured with different bile acids, including DCA and isoallolithocholic acid (isoalloLCA). Growth, hydrogen sulfide (H₂S) production, and virulence gene expression were measured. Notably, isoalloLCA was more potent in reducing growth, H …

Editorial: Parasitism: The Good, The Bad And The Ugly, Marco A. Juarez-Estrada, Danielle Graham, Xochitl Hernandez-Velasco, Guillermo Tellez-Isaias

Poultry Science Faculty Publications and Presentations

Editorial on the Research Topic

Parasitism: the good, the bad and the ugly

Ochrobactrum, Bacillus And Enterobacter Isolates Of Hot Water Spring Augment The Growth Of Zea Mays Seedlings, Noor-E-Saba Naz, Rida Batool, Nazia Jamil

Journal of Bioresource Management

Hot springs situated at high altitudes is a kind of remarkable ecosystem for the exploration of microbial flora. It was hypothesized that hot springs can harbor bacteria with plant growth-promoting and exopolysaccharides (EPS) producing ability that can favour the growth of plants. For the investigation of this hypothesis, seven EPS producing bacterial isolates were isolated from a water sample of hot water spring of Tattapani, Azad Kashmir and characterized morphologically and biochemically. Three out of seven isolates (BE1, BN1 and BN3) showed significant production of EPS (14-15 mg / 100 ml). Growth kinetics study revealed that optimum EPS production was …

A Review On Validation Of Enzyme Linked Immunosorbent Assay (Elisa) Techniques For Detection And Quantification Of Different Contaminant In Aquatic Environment, Sanwal Aslam, Zhen Zhang, Ali Muhammad

Journal of Bioresource Management

In world, contaminants and microbes are considered as serious problem in house water supplies as well as in aquatic environment for life. As a result, different countries are working to set level of these contaminants and microbes in domestic supply of water and in aquatic environment. In addition to these scientists are also working on reliable sampling and then their suitable analysis method for accuracy. It is also very important to improve and give justification for analytical techniques used in laboratories, which enable us to monitor level of various contaminants in aquatic environment. In this review, we have focused on …

Existing Evidence For Influenza B Virus Adaptations To Drive Replication In Humans As The Primary Host, Matthew J. Pekarek, Eric A. Weaver

Nebraska Center for Virology: Faculty Publications

Influenza B virus (IBV) is one of the two major types of influenza viruses that circulate each year. Unlike influenza A viruses, IBV does not harbor pandemic potential due to its lack of historical circulation in non-human hosts. Many studies and reviews have highlighted important factors for host determination of influenza A viruses. However, much less is known about the factors driving IBV replication in humans. We hypothesize that similar factors influence the host restriction of IBV. Here, we compile and review the current understanding of host factors crucial for the various stages of the IBV viral replication cycle. While …

Multiple Rsv Strains Infecting Hep-2 And A549 Cells Reveal Cell Line-Dependent Differences In Resistance To Rsv Infection, Anubama Rajan, Felipe-Andres Piedra, Letisha Aideyan, Trevor Mcbride, Matthew Robertson, Hannah L. Johnson, Gina Marie Aloisio, David Henke, Cristian Coarfa, Fabio Stossi, Vipin Kumar Menon, Harshavardhan Doddapaneni, Donna Marie Muzny, Sara Joan Javornik Cregeen, Kristi Louise Hoffman, Joseph Petrosino, Richard A. Gibbs, Vasanthi Avadhanula, Pedro A. Piedra

Research Symposium

Background: Respiratory syncytial virus (RSV) is the major viral driver of a global pediatric respiratory disease burden disproportionately borne by the poor¹. Thus, RSV, like SARS-CoV-2, combines with congenital and environmental and host-history-dependent factors to create a spectrum of disease with greatest severity most frequently occurring in those least able to procure treatment.

Methods: Here we apply whole genome sequencing and a suite of other molecular biological techniques to survey host-virus dynamics in infections of two distinct cell lines (HEp2 and A549) with four strains representative of known RSV genetic diversity.

Results: We observed non-gradient patterns of RSV …

Modeling Nonsegmented Negative-Strand Rna Virus (Nnsv) Transcription With Ejective Polymerase Collisions And Biased Diffusion, Felipe-Andres Piedra

Research Symposium

Background: The textbook model of NNSV transcription predicts a gene expression gradient. However, multiple studies show non-gradient gene expression patterns or data inconsistent with a simple gradient. Regarding the latter, several studies show a dramatic decrease in gene expression over the last two genes of the respiratory syncytial virus (RSV) genome (a highly studied NNSV). The textbook model cannot explain these phenomena.

Methods: Computational models of RSV and vesicular stomatitis virus (VSV – another highly studied NNSV) transcription were written in the Python programming language using the Scientific Python Development Environment. The model code is freely available on GitHub: …

Synthesis, Characterization, Antibacterial Activity, And Potential Water Filter Application Of Copper Oxide/Zeolite Composite, Elfa Aida Kamila, Zaenal Abidin, Irma Isnafia Arief, Trivadila Trivadila

Makara Journal of Science

Detection of E. coli bacteria in water samples can indicate the presence of other bacterial contamination in feces. Bacterial contamination can be treated with antibiotics, but excessive use of antibiotics leads to the emergence of multiresistant bacteria. Therefore, alternative antibacterial agents must be explored. Copper ion/zeolite composite has been widely studied as an antibacterial agent. However, the released copper ions accumulate in water and are toxic to humans; this can be prevented by converting copper ions into copper oxides (CuO and Cu₂O). In this study, copper oxide/zeolite composite is synthesized and characterized using the heating method, followed by …

Investigating The Impact Of Nonenzymatic Lysine Acetylation On The Function Of The Bacterial Ribosome, Sarah Caldwell Feid

Dissertations

An under-investigated target of lysine acetylation is the bacterial ribosome. Although lysine acetylations on the ribosome are common and conversed in diverse bacterial species, little work has been done to understand how lysine acetylation affects the bacterial ribosome. The goal of this work is to determine if lysine acetylation has functional impact on the bacterial ribosome. I have identified in vitro and in vivo effects of nonenzymatic, AcP-dependent lysine acetylation on translation and the ribosome. In vitro acetylation of transcription-translation reactions causes a translation-specific defect that is unaffected by the addition of the CobB deacetylase. This suggests certain AcP-sensitive residues …

Canine Testes Thin Sections Culture​, Nguyen Khoi Nguyen, Ramanpreet Singh

2023 Symposium

Canine testes thin section culture

Testes tissue culture systems would provide a tool to elucidate spermatogenesis mechanisms, with the aim of genetic preservation of mammals, especially endangered species. Our experiment aims to develop a culture system capable of producing viable mammalian sperm cells in vitro.

Dogs were chosen as the model organism because testes are readily available. Canine testes were obtained from a local veterinary clinic. Thin sections were generated using a commercial electric slicer. They then were cleaned using Dulbecco’s Phosphate-Buffered Saline (DPBS) supplemented with antibiotics then cultured in a modified Tissue Culture Medium 199 (TCM-199). Sections were …

Canine Testes Thin Sections Culture, Nguyen Khoi Nguyen, Ramanpreet Singh

2023 Symposium

Canine testes thin section culture

Testes tissue culture systems would provide a tool to elucidate spermatogenesis mechanisms, with the aim of genetic preservation of mammals, especially endangered species. Our experiment aims to develop a culture system capable of producing viable mammalian sperm cells in vitro.

Dogs were chosen as the model organism because testes are readily available. Canine testes were obtained from a local veterinary clinic. Thin sections were generated using a commercial electric slicer. They then were cleaned using Dulbecco’s Phosphate-Buffered Saline (DPBS) supplemented with antibiotics then cultured in a modified Tissue Culture Medium 199 (TCM-199). Sections were …

Development Of An Antibiotic Resistance Breaker To Combat Staphylococcus Aureus Pathogenesis Skin Infections Ssis A Multidisciplinary, Gopalakrishnan T Mr

Theses and Dissertations

Problem Statement: Staphylococcus aureus is a highly adaptable and tenacious pathogen known to cause a wide range of infections. The survival characteristics of the pathogen in different host niches are due to their ability to produce various pathogenic determinants. Although there lies the first line of treatment regimens with antibiotics, the pathogen defends at various levels to cause the waves of resistance - a silent pandemic “AMR”.

Notable strategies are through the protective layer, biofilm that enables the inter-/intra species communication (Quorum Sensing; QS) and emerges as a battleground to withstand host-immune response and antibiotic pressure. In parallel, several research …

What Is Microbial Dormancy?, Mark D. Mcdonald, Carlos Owusu-Ansah, Jared B. Ellenbogen, Zachary D. Malone, Michael P. Ricketts, Steve E. Frolking, Jessica Gilman Ernakovich, Michael Ibba, Sarah C. Bagby, J. L. Weissman

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Life can be stressful. One way to deal with stress is to simply wait it out. Microbes do this by entering a state of reduced activity and increased resistance commonly called ‘dormancy’. But what is dormancy? Different scientific disciplines emphasize distinct traits and phenotypic ranges in defining dormancy for their microbial species and system-specific questions of interest. Here, we propose a unified definition of microbial dormancy, using a broad framework to place earlier discipline-specific definitions in a new context. We then discuss how this new definition and framework may improve our ability to investigate dormancy using multi-omics tools. Finally, we …

Transcriptional Pausing Factor M1bp Regulates Cellular Homeostasis By Suppressing Autophagy And Apoptosis In Drosophila Eye, Anuradha Venkatakrishnan Chimata, Hannah Darnell, Akanksha Raj, Madhuri Kango-Singh

Biology Faculty Publications

During organogenesis cellular homeostasis plays a crucial role in patterning and growth. The role of promoter proximal pausing of RNA polymerase II, which regulates transcription of several developmental genes by GAGA factor or Motif 1 Binding Protein (M1BP), has not been fully understood in cellular homeostasis. Earlier, we reported that M1BP, a functional homolog of ZKSCAN3, regulates wingless (wg) and caspase-dependent cell death (apoptosis) in the Drosophila eye. Further, blocking apoptosis does not fully rescue the M1BP^RNAi phenotype of reduced eye. Therefore, we looked for other possible mechanism(s). In a forward genetic screen, members of the Jun-amino-terminal-(NH2)-Kinase (JNK) pathway …

Molecular Mimicry Of Noda Genes By An N-Acetylmuramoyl-L-Alanine Amidases Gene In A Nodule Forming Staphylococcus Cohnii [Research Note], Aberathne Herath Mudiyanselage Nadeeshani Ruwandika Aberathne, Wedage Methsala Madurangi Wedage, Dilantha Gunawardana

The Philippine Agricultural Scientist

A rare, nodule-forming, Gram-positive bacterium (Sub4) was isolated from the forage and cover crop Pueraria phaseoloides, which aligned at 99.26% sequence identity to a partial 16S rDNA sequence of Staphylococcus cohnii sp. This was the first record of a strain/species of Staphylococcus capable of independent, unassisted nodule formation in a legume host. When a nodA gene was sought by PCR using a pair of gene-specific primers synthesized using a related (Firmicutes) Paenibacillus sp. nodA gene, the reaction yielded a PCR product of similar size but a distinct identity. The resulting ~400bp PCR product coded for a likely N-acetylmuramoyl-L-alanine amidase …

Roost Selection By Synanthropic Bats In Rural Kenya: Implications For Human-Wildlife Conflict And Zoonotic Pathogen Spillover, Reilly T. Jackson, Paul W. Webala, Joseph G. Ogola, Tamika J. Lunn, Kristian M. Forbes

Biological Sciences Faculty Publications and Presentations

Many wildlife species are synanthropic and use structures built by humans, creating a high-risk interface for human–wildlife conflict and zoonotic pathogen spillover. However, studies that investigate features of urbanizing areas that attract or repel wildlife are currently lacking. We surveyed 85 buildings used by bats and 172 neighbouring buildings unused by bats (controls) in southeastern Kenya during 2021 and 2022 and evaluated the role of microclimate and structural attributes in building selection. We identified eight bat species using buildings, with over 25% of building roosts used concurrently by multiple species. Bats selected taller cement-walled buildings with higher water vapour pressure …

Effect Of Bovine Respiratory Disease On The Respiratory Microbiome: A Meta-Analysis, Samantha Howe, Beth Kegley, Jeremy Powell, Sicheng Chen, Jiangchao Zhao

Animal Science Faculty Publications and Presentations

Background: Bovine respiratory disease (BRD) is the most devastating disease affecting beef and dairy cattle producers in North America. An emerging area of interest is the respiratory microbiome’s relationship with BRD. However, results regarding the effect of BRD on respiratory microbiome diversity are conflicting.

Results: To examine the effect of BRD on the alpha diversity of the respiratory microbiome, a meta-analysis analyzing the relationship between the standardized mean difference (SMD) of three alpha diversity metrics (Shannon’s Diversity Index (Shannon), Chao1, and Observed features (OTUs, ASVs, species, and reads) and BRD was conducted. Our multi-level model found no difference in Chao1 …

Long-Term Impacts Of Conservation Pasture Management In Manuresheds On System-Level Microbiome And Antibiotic Resistance Genes, Mitiku Mihiret Seyoum, Amanda Ashworth, Kristina M. Feye, Steven C. Ricke, Phillip R. Owens, Philip A. Moore Jr., Mary Savin

Crop, Soil and Environmental Sciences Faculty Publications and Presentations

Animal manure improves soil fertility and organic carbon, but long-term deposition may contribute to antibiotic resistance genes (ARGs) entering the soil-water environment. Additionally, long-term impacts of applying animal manure to soil on the soil-water microbiome, a crucial factor in soil health and fertility, are not well understood. The aim of this study is to assess: (1) impacts of long-term conservation practices on the distribution of ARGs and microbial dynamics in soil, and runoff; and (2) associations between bacterial taxa, heavy metals, soil health indicators, and ARGs in manures, soils, and surface runoff in a study following 15 years of continuous …

Protocol For Optimizing Production And Quality Control Of Infective Ecohiv Virions, Hammodah Rawhi Hammodah Alfar, Daphne N. Pariser, Harry Chanzu, Smita Joshi, Daniёlle M. Coenen, Joshua Lykins, Kanakanagavalli Shravani Prakhya, Mary Jane Potash, Wei Chao, Jennifer Kelschenbach, David J. Volsky, Kelly Metcalf-Pate

Saha Cardiovascular Research Center Faculty Publications

EcoHIV is a model of HIV infection that recapitulates aspects of HIV-1 pathology in mice. However, there are limited published protocols to guide EcoHIV virion production. Here, we present a protocol for producing infective EcoHIV virions and essential quality controls. We describe steps for viral purification, titering, and multiple techniques to analyze infection efficacy. This protocol produces high infectivity in C57BL/6 mice which will aid investigators in generating preclinical data.

Interferon-Stimulated Gene Tdrd7 Interacts With Ampk And Inhibits Its Activation To Suppress Viral Replication And Pathogenesis, Sukanya Chakravarty, Gayatri Subramanian, Sonam Popli, Manoj Veleeparambil, Shumin Fan, Ritu Chakravarti, Saurabh Chattopadhyay

Markey Cancer Center Faculty Publications

Many viruses activate cellular autophagy in infected cells to facilitate their replication. Recently, we identified an interferon (IFN)-stimulated gene (ISG) Tudor domain containing 7 (TDRD7), which inhibits viral replication by blocking autophagy pathway. Here, we present a molecular mechanism for TDRD7 action and its relative contribution to protection against viral pathogenesis. TDRD7 inhibited the activation of adenosine monophosphate (AMP)-activated protein kinase (AMPK), a kinase required for initiating autophagy. Mechanistically, TDRD7 interacted directly with AMPK in the cytosolic compartment. Domain-mapping analyses revealed C-terminal Tudor domain of TDRD7 interacted with auto-inhibitory domain of AMPK. Deletion of Tudor domains abolished anti-AMPK and antiviral …

Past And Ongoing Field-Based Studies Of Myxomycetes, Steven L. Stephenson

Biological Sciences Faculty Publications and Presentations

Evidence from molecular studies indicates that myxomycetes (also called myxogastrids or plasmodial slime molds) have a long evolutionary history, and the oldest known fossil is from the mid-Cretaceous. However, they were not “discovered” until 1654, when a brief description and a woodcut depicting what is almost certainly the common species Lycogala epidendrum was published. First thought to be fungi, myxomycetes were not universally recognized as completely distinct until well into the twentieth century. Biodiversity surveys for the group being carried out over several years are relatively recent, with what is apparently the first example being carried out in the 1930s. …

The Bactericidal Activity Of A Novel Aneurinibacillus Aneurinilyticus Isolate Effectively Controls Foodborne Pathogens Campylobacter Jejuni And Listeria Monocytogenes, Peter M. Rubinelli, Rohana Liyanage, Jackson Lay, Jennifer C. Acuff

Chemistry & Biochemistry Faculty Publications and Presentations

Human zoonotic infection with Campylobacter is a major cause of gastroenteritis in the United States and worldwide. Listeria monocytogenes causes a potentially fatal infection in humans and is often attributed to contaminated food. Genetic typing has demonstrated that Campylobacter infection is often associated with the consumption of contaminated poultry products, with Campylobacter often colonizing the poultry gastrointestinal tract, while listeriosis is commonly associated with the consumption of contaminated ready-to-eat (RTE) foods. In this study, a strain of endospore-forming bacterium (strain NH) that is bactericidal towards the human food pathogens Campylobacter jejuni and Listeria monocytogenes was identified and characterized. Transwell experiments …

Aquatic Peptide: The Potential Anti-Cancer And Anti-Microbial Activity Of Ge18 Derived From Pathogenic Fungus Aphanomyces Invadans, Manikandan Velayutham, P. Snega Priya, Purabi Sarkar, Raghul Murugan, Bader O. Almutairi, Selvaraj Arokiyaraj, Zulhisyam Abdul Kari, Guillermo Tellez-Isaias, Ajay Guru, Jesu Arockiaraj

Poultry Science Faculty Publications and Presentations

Small molecules as well as peptide-based therapeutic approaches have attracted global interest due to their lower or no toxicity in nature, and their potential in addressing several health complications including immune diseases, cardiovascular diseases, metabolic disorders, osteoporosis and cancer. This study proposed a peptide, GE18 of subtilisin-like peptidase from the virulence factor of aquatic pathogenic fungus Aphanomyces invadans, which elicits anti-cancer and anti-microbial activities. To understand the potential GE18 peptide-induced biological effects, an in silico analysis, in vitro (L6 cells) and in vivo toxicity assays (using zebrafish embryo), in vitro anti-cancer assays and anti-microbial assays were performed. The outcomes of …

Evaluation Of Organic And Synthetic Herbicides Applications On Indicators Of Soil Health And Weed Suppression In A Conventional Cropping System In Louisiana, Caitlin Denux

LSU Doctoral Dissertations

Synthetic herbicides, with their varying modes of action, are well known for their efficiency in the suppression and control of weed species in U.S. agriculture. However, the consequences of using synthetic herbicides without attention to the surrounding environment can produce chemical run-off, changes in soil health and soil health conditions, and create herbicide-resistant weeds. These consequences have encouraged growers to find alternatives in their weed management programs. In this study, organic and synthetic herbicides were assessed on how well they performed in weed suppression surveys, and their potential effects on soil health indicators. Commonly used indicators include soil pH, β-glucosidase …

Identification Of A Small Regulatory Rna Usps Associated With The Universal Stress Protein In Lactobacillus Species, Zarah M. Fowler, Sasha S. Bronovitskiy, Finn K. Rose, Brian M. Lee

Journal of the South Carolina Academy of Science

The gut microbiome is a complex habitat with many bacterial species, each playing crucial roles in regulating various physiological processes in the body. As the use of probiotics to combat human disease continues to increase, it is important to understand the mechanisms by which probiotic bacteria regulate their interactions with other bacteria and their host. Our exploration of the physiological functions of probiotic bacteria hopes to elucidate the role of small regulatory RNA (sRNA) in regulating gene expression within the microbiome. The goal of this project was to characterize the structure and function of the sRNA, UspS, which is found …

Celebrating Leeuwenhoek’S Life 300 Years Later, Alan L. Gillen

Faculty Publications and Presentations

Antony van Leeuwenhoek entered eternity 300 years ago (August 26, 1723). He made hundreds of discoveries with his simple microscope. Perhaps his most notable discovery was of bacteria and its many shapes. He found great joy in God [http://answersingenesis.org/god/] ’s smallest creatures. I believe that his curiosity about the world, fueled by his belief in biblical creation [http://answersingenesis.org/creation/] , led to his discovery of bacteria. In a similar manner, his curiosity of blood led to his discovery of red corpuscles and capillaries. He remained a curious man until the last days of his life 300 years …

Sero-Epidemiology Of Rotavirus Gastroenteritis In Children In Ilorin, Kwara State, Omosigho Omoruyi Pius, Izevbuwa Osazee Ekundayo, Saheed Ibrahim Damilare

Journal of Bioresource Management

Rotavirus is responsible for the most severe dehydrating diarrhea among young children due to gastroenteritis. In this study, we aimed to ascertain the occurrence of childhood gastroenteritis caused by Rotavirus among infants and young children who are younger than 5 years of age in Ilorin, Kwara State and determined the risk factors posing the challenges to be susceptible to diarrhea associated with rotavirus in Ilorin, Kwara State. Diarrhea stool samples were collected from children who passed watery stools, who met predetermined inclusion criteria and who presented at the study hospitals Viz: General Hospital and Specialist Hospital Alagbado and Children Specialist …

Additive Effects Of Cyclic Peptide [R4w4] When Added Alongside Azithromycin And Rifampicin Against Mycobacterium Avium Infection, Melissa Kelley, Kayvan Sasaninia, Arbi Abnousian, Ali Badaoui, James Owens, Abrianna Beever, Nala Kachour, Rakesh Kumar Tiwari, Vishwanath Venketaraman

Pharmacy Faculty Articles and Research

Mycobacterium avium (M. avium), a type of nontuberculous mycobacteria (NTM), poses a risk for pulmonary infections and disseminated infections in immunocompromised individuals. Conventional treatment consists of a 12-month regimen of the first-line antibiotics rifampicin and azithromycin. However, the treatment duration and low antibiotic tolerability present challenges in the treatment of M. avium infection. Furthermore, the emergence of multidrug-resistant mycobacterium strains prompts a need for novel treatments against M. avium infection. This study aims to test the efficacy of a novel antimicrobial peptide, cyclic [R4W4], alongside the first-line antibiotics azithromycin and rifampicin in reducing M. avium survival. Colony-forming unit (CFU) …

Investigating Putative Toxin-Immunity Pairs From A New Class Of Rhs Proteins In Pseudomonas Aeruginosa, Syed Faieq Imam

Master's Theses

Pseudomonas aeruginosa is a gram-negative, opportunistic pathogen that is ubiquitous in nature, especially in human/animal impacted environments. One of the reasons for this is the arsenal of protein systems and toxins that P. aeruginosa utilizes to mold its environment to suit its needs. Recently, we discovered a novel locus in the P. aeruginosa genome, which resembles other loci that are known to mediate interbacterial competition. Our bioinformatic analysis found this locus to be enriched in a rearrangement hotspot (rhs) motif which led us to ad-interim term the three ORFs of the loci as rhsB, rhsC, and rhsI. Furthermore, we have …

Mutation-Induced Changes In The Stability, B-Cell Epitope, And Antigenicity Of The Sars-Cov-2 Variant Spike Protein: A Comparative Computational Stud, Nira Meirita Wijayanti, Muhammad Hermawan Widyananda, Lailil Muflikhah, Nashi Widodo

Karbala International Journal of Modern Science

The spike (S) protein is a major antigenicity site that targets neutralizing antibodies and drugs. The growing number of S protein mutations has become a severe problem for developing effective vaccines. Here, we investigated four severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants that were the most infectious and widespread during the COVID-19 pandemic to determine the trends and patterns of mutation-induced changes in the stability, B-cell epitope, and antigenicity of the SARS-CoV-2 S protein. The data showed that the Beta and Gamma variants had three mutations on the receptor-binding domain (RBD), which is the specific site on the S …