Loss of Bifidobacteria in Lyme Disease: Cause or Effect

Introduction: Lyme disease is a poorly understood condition which starts with a rash but may continue with chronic fatigue and neurological symptoms. Approximately 1 in 5 early Lyme disease patients have GI symptoms, such as nausea, anorexia, abdominal pain, or diarrhea. Lyme disease is thought to be cased by microbes in the spirochetes phylum transmitted by black legged ticks. Lyme-related healthcare costs in America exceed 1.3 billion dollars annually. Bifidobacteria are known for their beneficial probiotic actions within the human gut microbiome. Their numbers are reduced in severe COVID-19, Clostridioides difficile infection and Inflammatory Bowel Disease. To our knowledge Bifidobacteria levels have not been studied in Lyme disease patients. Given the importance of Bifidobacterium abundance in other diseases, we focused on relative abundance of Bifidobacterium in fecal samples of patients with Lyme disease compared to controls. Method(s): Fecal samples were assessed for relative abundance of Bifidobacterium in Healthy Control subjects without Lyme disease (n=20) compared to patients with Lyme disease (n=39). The average symptom duration in patients with Lyme disease was 5 years and none were on antibiotics 2 weeks prior to sample collection (range of symptoms from 1 month to 20 years, all treated initially with antibiotics).Metagenomics Next Generation sequencing was performed on fecal samples, where DNA samples were extracted and normalized for library downstream analysis using Shotgun Methodology. Mann- Whitney Statistical test was used for comparison. This study was IRB approved. Result(s): Relative Abundance of bifidobacteria was significantly decreased (p< 0.0001) in patients with Lyme disease. Median and interquartile range (IQR) were: Control (Median:4.175%;IQR:1.72-10.27%) and Lyme disease (Median:0.0014%;IQR:0.00%-0.96%)(Figure). 30/39 Lyme disease patients (77%) were found to possess < 1% relative abundance of Bifidobacterium in their stool sample. Of interest only 1/39 samples showed presence of Spirochetes in stool samples. Conclusion(s): This is the first study that demonstrates low levels of Bifidobacteria in patients with chronic Lyme disease. These results raise three questions;whether the disease was caused by 1. the original microbe creating loss of Bifidobacterium 2. baseline low Bifidobacteria due likely to either diet or medications or 3. excessive treatment. Given Lyme disease comprises a gut dysbiosis issue, therapies should also aim at restoration of depleted Bifidobacteria. (Figure Presented).

Prevotella differentially regulates the inflammatory response of human monocytes to SARS-CoV-2 spike glycoproteins

Patients suffering severe COVID-19 show an aggressive and excessive immune response against the SARS-CoV-2 coronavirus, known as a cytokine storm. If left untreated these patients face the risk of tissue damage, multi-organ failure and death. A high relative abundance of Prevotella copri has been reported in patients with newly diagnosed rheumatoid arthritis (RA). On the other hand, it has been observed that Prevotella histicola can modulate the inflammatory manifestations of autoimmune diseases like multiple sclerosis, and it is now being evaluated as a monoclonal microbial treatment in COVID-19. We observed that pre-treatment with P. histicola decreased NF-kB activation, while pre-treatment with P. histicola and P. copri decreased IRF activation in monocytes upon SARS-CoV-2 glycoprotein. Our findings suggest that exposure of blood immune cells, such as monocytes, to commensal species of Prevotella may reduce the inflammatory response to SARS-CoV-2 glycoprotein. Besides treatments targeting the viral infection, other treatments such as immunomodulation by bacteria aiming to reduce or regulate the inflammatory process in COVID-19 to avoid the development of related complications may be considered.

Airway microbiota profiles associated with total lung capacity after severe COVID-19

Background: SARS-CoV-2 infection induces disturbed airway microbiota during the acute phase of infection that may contribute to persistence of long-term pulmonary sequelae. To date it is unclear if the presence of a disrupted microbiota following severe disease is linked to long-term pulmonary function impairment. This one-year follow-up study investigated the association between airway microbiota and lung function after severe COVID-19. Method(s): In the Swiss COVID Lung study (NCT04581135), we conducted 16S rRNA sequencing on upper respiratory tract specimen obtained by oropharyngeal swabs 3 to 12 months after hospitalisation from 72 subjects (total samples n = 169) with severe COVID-19. Subjects underwent 1 - 3 follow-up visits during which lung function testing was performed to investigate correlation with the richness and composition of airway microbiota. Result(s): Total lung capacity (TLC) was negatively correlated with bacterial richness (p = 0.0081). Recovered COVID-19 subjects with ongoing respiratory impairment (TLC < 80%) showed low phylum heterogeneity with a majority of the dominant taxa being Bacteroidetes (64% of the 50 most abundant taxa in the group). In contrast, the phylum with the largest number of dominant taxa in subjects with TLC >= 80% was Firmicutes (48% of the 50 most abundant taxa in the group). Conclusion(s): Patients with impaired total lung capacity between 3 and 12 months after severe COVID-19 have a distinct oropharyngeal microbiota from those with restored total lung capacity. Future studies need to assess the contribution of microbiota to lung function impairment after severe COVID-19, as airway microbiota analysis may assist monitoring of sequelae and recovery.

A Role of Black Fungus in COVID-19: A Review

Mucormycosis is a rare type of fungal infection commonly known as zygomycosis, the infection tends to crop up more commonly in individuals with low and weakened immunity level, if left untreated, the mucormycosis can be life-threatening and fatal. Mucormycosis previously known as zygomycosis is a consequential type of infection caused by several mildews known as micromycetes. The revised taxonomical studies revealed that the micromycetes causing the infections are classified as the species of phylum Glomeromycota, class Glomeromycetes, subphylum Mucoromycotina, order Mucorales. The genera of Rhizopus, Mucor, Lichtheimia, Cunninghamella, Rhizomucor, and Apophysomyces, constitute the causative agents of the majority of cases of mucormycosis. The angioinvasive type of disorder caused by mucormycosis is further classified as Mucorales. The patients with Diabetes ketoacidosis and diabetes mellitus are at high-risk factors, followed by the patients with organ transplant, immunocompromised disease, and malignancy. The route of exposure to Mucormycosis may be through the wounded infection that can be pneumonic, or dermal in origin. In the ectodermal form, the fungal organism can invade the skin through open or puncture wounds, or the laceration on the skin. However, the infection has a high mortality rate, the key to successful treatment is early diagnosis, and administration of antifungal drugs, with extensive therapy, followed by surgical debridement of the infection. The morbidity and mortality rate are still at a high number, due to the negligence of the patient to seek medical treatment. Hence the early diagnosis and treatment with antifungal drugs with surgical debridement is a must. The efficacy of oral and venous formulations in the treatment of mucorales is still under debate. Despite the aggressive therapy, the mortality rate is increasing worldwide. The studies have to be conducted to invent the fastest treatment protocol for the treatment of Mucormycosis.

We don't know everything (we know) about dogs

This article highlights information on the beneficial and pathogenic microorganisms of the oral cavity of dogs, effects of grass consumption, and the breeding and care of dogs.

COVID-19: DIARRHEA, INFLAMMATION AND INTESTINAL MICROBIOTA

Introduction: Increased inflammatory cytokines has been observed in COVID-19 patients and there is evidence showing an alteration in gut-microbiota composition. SARS-CoV-2 can cause gastrointestinal symptoms, such as diarrhea. Evidence of an altered gut-microbiota composition and cytokines levels in COVID-19 diarrhea patients is lacking. Objectives: To compare serum cytokine levels and gut microbiota between COVID-19 diarrhea (D-COVID- 19) and non-diarrhea (NonD-COVID-19) patients and non- COVID-19 controls (HC). Material and methods: We included 143 hospitalized COVID-19 patients (positive quantitative reverse transcription PCR) in a single University Hospital, and 53 ambulatory HC (negative rapid serological test) were included. Blood and stool samples were collected at hospital admission in COVID-19 patients and at the time of HC recruitment. 27- pro and anti-inflammatory cytokines (Bio-Plex Pro™, Bio- Rad) were measured. Gut microbiota composition and diversity profiles were characterized by sequencing the 16S rRNA gene V3-V4 region amplified using DNA extracted from stool samples. Bioinformatics analysis was performed with QIIME2 software. First, we compare cytokine levels between COVID- 19 and HC and then COVID-19 with and without diarrhea. All comparisons were adjusted for age, sex, and BMI with linear regression. Results: The mean age in COVID-19 patients was 54 +/- 15 years (F=50%) and 52 +/- 8 (F=62%) for HC. Diarrhea was present in 19 (13.29%) of COVID-19 patients. COVID-19 patients had significative higher levels of: IL- 1ra, IL-2, IL-6, IL-7, IL-8, IL-13, IP-10 and PDGF-bb. Significant lower values of: IL-9, FGF -basic, MIP-1β, TNF-α were observed in D-COVID-19 compared to NonD-COVID-19. COVID-19 patients had a significant reduction of bacterial species (p=0.0001), and diversity and complexity of the bacterial community (Shannon's index) (p=0.0001) compared to the HC. There was no difference between D-COVID-19 and NonD-COVID-19. There were also changes in the composition of the microbiota associated with COVID-19. At the phylum level, COVID-19 patients showed a significant decrease in Actinobacteria and Firmicutes, and an increase in Bacteroidetes. At species level, an increase of 4 species of the genus Bacteroides was observed in COVID-19 patients. 31 very diverse bacterial species were found, all decreased in D-COVID-19. Conclusions: An alteration in serum cytokine levels was observed between COVID-19 and HC. D-COVID-19 had a decrease in some proinflammatory cytokines. A significant decrease in richness and species diversity of gutmicrobiota was observed in COVID-19 patients compared to HC, but no significant differences were observed between D-COVID-19 and NonD-COVID-19. However, in D-COVID- 19, a decrease in some bacterial species was observed.(Table Presented)(Figure Presented)

MARKED CHANGES ARE SEEN IN THE SMALL BOWEL MICROBIOME AS A RESULT OF THE COVID-19 PANDEMIC

SARS-CoV-2 has had a profound impact on the human population in the last 24 months. This includes dramatic changes in lifestyle, hygiene, and altered food sources/consumption patterns, which could directly impact the small bowel microbiome on an individual and perhaps even at a population level. To date, this has not been examined. Here, we compare small bowel microbial profiles in subjects before SARS-CoV-2 and intra-pandemic. Methods: The REIMAGINE study is a large-scale study using validated methods for duodenal aspirate collection and microbiome sequencing in subjects undergoing upper endoscopy. Subjects were divided into 2 groups: pre-pandemic, from February 2019 to March 2020;and intrapandemic, from April 2021 to September 2021. Groups were matched for gender, age, and BMI. Duodenal aspirates were collected, and microbial DNA was isolated using the MagAttract PowerSoilDNA Kit. V3 and V4 libraries were sequenced on a MiSeq. Reference-based Operational Taxonomic Unit clustering was performed using SILVA v132 database. Taxonomic analysis was performed with CLC Microbial Genomics Module v.2.5 and MicrobiomeAnalyst, and duodenal microbial alpha- and beta-diversity indices were calculated. Significance was determined by Wilcox test. Results: In total 94 subjects were included in the analysis. The overall duodenal microbiome profile (beta-diversity) of intra-pandemic subjects (n=38, mean age= 51 ± 18, mean BMI =23.9 ± 4.7) was significantly different from pre-pandemic subjects (n=56, mean age= 51 ± 15, mean BMI = 25.24 ± 4.9)(p<0.002, Fig1A), with no significant changes in duodenal microbial alpha diversity between groups (Fig1B). Significant duodenal microbial taxonomic differences were identified between groups, including changes in the relative abundance (RA) of 2 phyla, 3 classes, 6 orders, 4 families and 23 genera (Fig2A). At the phylum level, Actinobacteria RA was significantly decreased in the intra vs. the pre group (FC=-1.99, P=9.83E-8, Fig2B). Additionally, at the genus level, RA of Rothia (P= 6.85E-7), Pseudomonas (P=0.0376), and Escherichia (P=0.0092) were significantly decreased in the intra group (Fig2A). Of note, the phylum Deinococcus (P=0.0016) was increased in the intra vs. the pre group (Fig2B). Conclusion: In this first study examining the effect of the COVID-19 pandemic on the small bowel microbiome, we show substantial changes in microbial profiles intra-pandemic as compared to pre-pandemic. The duodenal microbiome of intra-pandemic subjects was associated with less disrupter bacteria (Escherichia and Pseudomonas), commonly associated with GI disorders. In contrast, Deinococcus phylum was increased intra-pandemic. This phylum includes organisms resistant to sanitation and increased in the nasal passage of people during the pandemic. The short and long term impact of these changes on human health require further study.(Figure Presented)

Assessment of wastewater and microbiological characteristics from selected hospitals in Umuahia metropolis

There is an increasing universal awareness of environmental problems arising as a result of COVID- 19 pandemic and pollution especially in Nigeria. Among the source of this problem is effluent discharge from industries, particularly hospitals in arable farmlands and environs. Two outstanding hospital were purposively selected;Madonna Catholic Hospital and Abia Specialist Hospital in Umuahia, Abia State. Their wastewater samples were collected from three different wards;maternity, general private, and general out-patients department (GOPD) wards. Results obtained show significant variation in physiochemical properties in some wards and heavy metals across all wards. Seven bacteria species;Staphylococcus aureus, Escherichia coli, Klesbsiella pneumonia, Pseudomonas aeruginosa, Proteus vulgaris, Bacteriod sp and Streptococcus pyogenes and one fungi specie- Candida albican were recorded from the samples. The bacterial load in Madonna ranged from 209.04 to 232.95cfu/ml in January, February, and March each and was statistically the same in the three wards (p>0.05). Fungi load ranged from 1.58 to 2.35cfu/ml in January, February, and March each and also significantly different at (p>0.05). The frequency of microbial characteristics isolated in the two hospital wastewater ranged from 33 to 100% with 100% of Staphylococcus aureus and Escherichia coli in all the wards, while other species varied significantly between 67 and 33% each. The results of the isolated bacteria from hospital wastewater showed resistivity to the tested antibiotics, and as therapeutic agents. Therefore, results call for need for urgent attention to be given to the discharge of wastewater from hospitals to ensure that food production around the environment is not contaminated.

Effects of recombinant porcine Lactobacillus reuteri secreting expression of bovine lactoferrin peptides on growth performance of newborn piglets and protective effect against diarrhea virus infection

This experiment was conducted to investigate the effects of recombinant porcine Lactobacillus reuteri secreting bovine lactoferrin peptide (LFCA) on growth performance of newborn piglets and the protective effect on porcine transmissible gastroenteritis virus (TGEV)infection which caused piglet diarrhea. Experiment 1:thirty-six one-day-old newborn piglets with an average body weight of about 1.5 kg were randomly divided into 3 groups, which were pPG-LFCA/LR-CO21 group, pPG/LR-CO21 group and control group, each group with 12 piglets. Piglets in each group were orally administered recombinant porcine Lactobacillus reuteri expressing LFCA pPG-LFCA/LR-CO21, containing empty vector plasmid PPG/LR-CO21 and equal volume phosphate buffer (PBS);oral administration continued for 3 days, and the observation time after oral administration was 14 d. During the period, piglets were fed freely, and the changes of body weight and diarrhea were recorded. Experiment 2:thirty one-day-old newborn piglets with an average body weight of about 1.5 kg were randomly divided into 5 groups and given TGEV with a half tissue culture infection dose (TCID50) of 10-7.50/mL by oral administration of 1, 3, 6, 9 and 12 mL, respectively. The observation period of 7 d was set to analyze the conditions of half lethal dose. Experiment 3:another thirty-two newborn piglets with an average body weight of about 1.5 kg were selected as experimental animals and randomly divided into 4 groups, with 8 piglets in each group. The groups were pPG-LFCA/LR-CO21 group, pPG/LR-CO21 group, control group and TGEV infect group. There were 8 replicates in each group and 1 piglet in each replicate. Each head of the experimental group was orally fed ppG-LFCA/LR-CO21, pPG/LR-CO21 and equal volume of PBS at a dose of 2..1010 CFU per day for 1 consecutive week. At 8 days of age, TGEV was infected by oral administration at half lethal dose, and samples were collected after 7 days of infection. The weight change and diarrhea of each group of piglets were recorded;hematoxylin-eosin staining was used to detect the length of intestinal villi and the depth of crypts;enzyme linked immunosorbent assay (ELISA) was used to determine total serum total immunoglobulin G (IgG) and total secretory immunoglobulin A (sIgA) antibody contents. RT-qPCR was used to detect the mRNA relative expression levels of Claudin-1, Occludin, tight junction protein-1 (ZO-1), inflammatory cytokines interleukin-6 (IL-6), interleukin-8 (IL-8), interferon-P (IFN-P), tumor necrosis factor-a (TNF-a) and Toll-like receptor 2 (TLR2). The flora structure of the contents of the piglet's cecum was analyzed. After oral recombinant porcine Lactobacillusreuteri, compared with the control group, the average daily gain of newborn piglets in the pPG-LFCA/LR-CO21 group was significantly increased (P < 0.01), while the diarrhea rate was significantly decreased (P < 0.01). Compared with TGEV infection group, the average daily gain of piglets in pPG-LFCA/LR-CO21 group was increased and diarrhea rate was decreased, and the differences were significant (P < 0.05). Villus height and the ratio of villus height to crypt depth in jejunum and ileum were significantly increased (P < 0.05). The contents of total IgG and intestinal mucosal total sIgA antibody in serum of piglets were significantly increased (P < 0.05);the mRNA relative expression levels of tight junction protein-related genes Claudin-1, Occludin and ZO-1 in intestinal mucosal tissue were extremely significantly increased (P < 0.01), and the serum TNF-a content was extremely significantly decreased (P < 0.01). Serum IFN-P, IL-6, IL-8 and TLR2 contents were significantly increased (P < 0.01), and the survival rate of piglets was improved. The analysis of the bacterial diversity in the contents of the piglets' cecum showed that the proportion of normal intestinal flora of piglets decreased after TGEV infection. Compared with the TGEV infect group, the proportion of pathogenic bacteria Bacteroides in piglet's intestinal flora decreased by o