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• Bentonite clay Protects Intestinal Normal Flora and Alleviates Pseudomembranous Colitis following antibiotic usage. • Bentonite clay Adsorbs Residual Antibiotics in the gut, without affecting concentration of the antibiotics in the blood. • Bentonite clay possesses antibacterial properties against pathogens and reduces the concentration of bacterial toxins in the intestine. • Bentonite clay reduces the germination of Clostridium's spores by absorbing bile salts and inhibits bacterial proteases by absorbing zinc. • Bentonite clay improves pseudomembranous colitis through immunomodulation and wound healing. As the most commonly used drugs, antibiotics have several adverse effects. When intravenous broad-spectrum antibiotics are administered for infections outside the gut, they are also released into the gut, upsetting the natural balance of flora and potentially causing opportunistic infections there. Pseudomembranous colitis induced by the excessive growth of Clostridioides difficile following antibiotic administration and disruption of the normal gut flora is a main consequence of antibiotic therapy. Unfortunately, effective strategies to mitigate the damage caused by antibiotics to the microbiota remain elusive, with limited information available on the subject. Several studies have examined the effects of enzymatic inactivation of specific antibiotics, but non-enzymatic inactivation of antibiotics has been explored to a lesser extent, resulting in the introduction of only a restricted number of products. We believe that bentonite clay, an underutilized substance that exhibits promising features for counteracting the unfavorable effects of antibiotics has a high potential to circumvent the side effects of antibiotics. It is a cost-effective and safe and has garnered significant attention in the medical history, predominantly for its superior adsorption properties. Given the diverse characteristics of bentonite clay, capacity to protect the natural flora, antibacterial and anti-inflammatory traits, and its contribution to wound healing, it is plausible to assume that bentonite clay exhibits significant potential in mitigating the detrimental consequences of antibiotic on the natural gut microbiota and following pseudomembranous colitis. Further research is necessary to unravel the full extent of its therapeutic effects in combating this condition. [ABSTRACT FROM AUTHOR]

This study was undertaken to investigate the bacterial and fungal microflora on the external genitalia of a population of healthy male donkeys in the state of Michigan, USA. The aim was to identify and determine the frequency of occurrence of these microorganisms using seven different isolation media and standard microbiological procedures. The sites (urethral fossa [fossa glandis], dorsal diverticulum of the urethral sinus, distal urethra, and penile surface) in the distal reproductive tract were cultured and each isolated microorganism identified. Ten different genera of gram-positive bacteria, eight different genera of gram-negative bacteria, and two genera of fungi were isolated from the external genitalia of the 43 donkeys in this study. All 43 donkeys yielded gram-positive bacteria (2–8 species) from all four sites sampled. Arcanobacterium spp., Corynebacterium spp., and Bacillus spp. were the most frequently isolated gram-positive bacteria. Gram-negative bacteria were cultured from 16 (37.2%) of the 43 donkeys, with Acinetobacter lwoffii (16.3%), Oligella urethralis (11.6%), and Taylorella asinigenitalis (9.3%), the most frequently isolated. Fungi were cultured from only 5 (11.6%) of the 43 donkeys, with Rhizopus spp. isolated from 3 (7.0%) and Cladosporium spp. from 2 (4.7%) individuals. The testes and epididymides collected from 40 donkeys at time of castration were culture negative. Few differences were found in the bacterial flora between prepubertal and mature intact and castrated donkeys. Of notable interest was the scarcity of known equine pathogens across the population tested and isolation of T. asinigenitalis from normal donkeys, especially prepubertal individuals and previously castrated males. [ABSTRACT FROM AUTHOR]

Abstract: This study was undertaken to investigate the prevalence of Taylorella asinigenitalis in a subset of the donkey population of Michigan and in other equids on farms on which the organism was identified. Other aims were to further characterize the carrier state in terms of persistence and preferred sites of colonization of T. asinigenitalis in the male donkey as well as determine the genotype of any isolates of the organism. Initial testing of 43 donkeys and 1 mule turned up 4 (9.3%) donkeys culture positive for T. asinigenitalis. The 4 culture-positive donkeys resided on 2 farms accommodating a collective total of 89 equids, of which 23 (25.8%) were confirmed positive for T. asinigenitalis. The positive equid population on the 2 farms comprised 14 (67%) of 21 gelded donkeys, 8 (36.4%) of 22 intact male donkeys, and 1 (25%) of 4 gelded horses. T. asinigenitalis was not isolated from 27 female donkeys, 11 female horses, 2 female mules, 1 male horse, or 1 male mule resident on these premises. Isolations of the bacterium were obtained from a number of male donkeys whenever they were sampled over a span of 33 months; preferential sites of isolation were the urethral fossa (fossa glandis), dorsal diverticulum of the urethral sinus, and terminal urethra. Isolates of T. asinigenitalis from the 23 culture-positive equids comprised 2 genotypes, one identical to the type strain isolated in California in 1997, and the other identical to 2 strains isolated from donkey jacks in Kentucky in 1998. [Copyright &y& Elsevier]

Clin Microbiol Infect 2012; 18: 497-501 Abstract Phage lysin was evaluated as a substitute for antibiotics in sputum samples processed by a modified Petroff's method for the detection of Mycobacterium tuberculosis with the MGIT 960 system. One hundred and fifty sputum samples were processed, inoculated onto two slopes of Lowenstein-Jensen medium, and divided in to two aliquots of 0.5 mL each. One aliquot was added to 7 mL of MGIT medium containing polymyxin B, amphotericin B, nalidixic acid, trimethoprim and azlocillin (PANTA) (MGIT-PANTA) and the other was added to 7 mL of MGIT medium containing 0.8 mL of lysin (MGIT-Lysin). The samples were randomized and incubated at 37°C in the MGIT 960 system. The sensitivity and specificity of MGIT-Lysin were 97% and 88%, respectively, as compared with MGIT-PANTA. The average times to detection with MGIT-Lysin and MGIT-PANTA were 9.3 and 8.6 days, respectively. The rate of contamination with MGIT-PANTA and MGIT-Lysin were 16% and 7.3%, respectively. Phage lysin can be substituted for antibiotics in processed sputum samples for the detection of M. tuberculosis. [ABSTRACT FROM AUTHOR]

Genetically modified Lactococcus lactis (L. lactis), a probiotic bacterium, able to secrete β-lactamase (29 kDa), was used as a vector for the oral delivery of β-lactamase to the rats. Three different doses of L. lactis were administered to the rats, and the resulted β-lactamase oral bioavailability was studied, and compared to the solution form. The oral administration of 1.2 × 107, 3 × 107, and 8 × 107 colony-forming units of L. lactis led to 145, 209, and 364 mU of β-lactamase absorbed, and the corresponding bioavailability was 8.7%, 15.5%, and 20.8% based on the in vitro production of β-lactamase by L. lactis. The oral administration of 504 mU and 1008 mU β-lactamase free solution resulted in 30 and 47 mU absorbed, a bioavailability of 5.9% and 4.7%, respectively. L. lactis significantly (p < 0.01) increased the oral bioavailability compared to the free solution form. A significant (p < 0.01) increase in the MAT value as compared to the solution, demonstrated that L. lactis can be used as a sustained delivery system. In conclusion, there is a linear relationship between L. lactis dose and these absorption PK parameters within L. lactis dose range of the current study. © 2009 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 98:2573–2580, 2009 [ABSTRACT FROM AUTHOR]

Abstract: The mechanical pressure exerted during centrifugation and the chemical pressure experienced when sputum specimens are processed, leave the tubercle bacilli in the sputum unsuitable for rapid detection especially in phage based assays. Thus, growing Mycobacterium tuberculosis in broth, at least overnight, is mandatory for allowing the tubercle bacilli to recoup. During this time the surviving colonizing flora grow faster and overgrow tubercle bacilli interfering with TB diagnosis. In the present study normal flora surviving the action of 4% NaOH was isolated and characterized. Phages capable of killing 14 different species representing this normal flora were isolated from soil and sewage samples and characterized. A novel and bio-friendly approach to treat sputum samples with a cocktail of three phages capable of killing most of the 14 representative organisms and not infecting mycobacteria is explored to control the overgrowth of colonizing bacteria in broth culture. While 26 of the 100 sputum samples processed by modified Petroff''s procedure showed growth of colonizing flora on blood agar, all of them when grown in broth overnight showed mixed, confluent growth. The addition of phagebiotics controlled them all, showing a significant reduction in colony forming units but resulting in few discrete colonies in 54 samples. Isolation of phages capable of controlling these surviving organisms and including them in the phagebiotics mixture should lead to the control of colonizing bacteria effectively. [Copyright &y& Elsevier]

Abstract: Lactococcus lactis subsp. lactis transformed with Plasmid ss80 (encoding the production and secretion of TEM β-lactamase) was used for the delivery of β-lactamase through the C-33A (cervix cell) monolayer. The viability of the cell monolayers co-cultured with L. lactis was examined by the trypan blue exclusion method. The integrity of the monolayers was monitored by measuring the transport of mannitol and propranolol as well as the transepithelial electrical resistance. The transport rate of β-lactamase through C-33A monolayer was increased by four- and nine-folds (p <0.05) at the first hour by the transformed L. lactis compared to the free solution with or without presence of the untransformed L. lactis, respectively. This increase was gradually diminished after the 1st hour: it became 30 and 50% (p <0.05) at 10h. The presence of the untransformed L. lactis with free solution delivery also increased the transport rate by 100% at 1h (p <0.05) and 15% at 10h (p >0.05). The increase in transport rate by the transformed L. lactis is most probably due to the concentrate of β-lactamase on C-33A monolayer. When co-cultured with the L. lactis, the C-33A cell viability and the monolayer TEER remained steady for 10h. The presence of L. lactis did not change the transport of propranolol and mannitol through the monolayers. In conclusion, the transformed L. lactis significantly (p <0.05) increased the transport of β-lactamase through the cervical monolayers, indicating probiotic bacteria delivery may be a promising approach for protein delivery through the vagina. [Copyright &y& Elsevier]

Abstract: The objective was to use normal flora to deliver protein/peptide drugs orally. A probiotic bacterium, Lactococcus lactis subsp. lactis (L. lactis) transformed with Plasmid ss80, which made it able to synthesize and secrete β-lactamase, a 29kDa protein, was used as the delivery system for β-lactamase. Oral absorption of β-lactamase in rats when delivered by this L. lactis system was investigated. The oral bioavailability of β-lactamase delivered by 3×107 of the L. lactis was equivalent to 209mU of i.v. dose, and the estimated relative bioavailability was 16.7%. When delivered by β-lactamase free solution form, the relative oral bioavailability was 4.7%, which increased to 6.0% when co-administered with 3×107 of the untransformed L. lactis. The results demonstrated that the L. lactis significantly increased the β-lactamase oral bioavailability by 2–3-folds (p <0.01), the mean residence time (MRT) by 3–4 times (p <0.01), and the mean absorption time (MAT) by 6–14 times (p <0.01), as compared to the free solution form with/without the untransformed L. lactis. In conclusion, the L. lactis is more efficient in delivering β-lactamase orally compared with the free solution form. It also provides a sustained delivery mechanism for β-lactamase. Gene-transformed normal flora may be used as an efficient and sustained delivery system for protein drugs through oral route. [Copyright &y& Elsevier]

Summary: Halibut, the largest of all flatfishes is a valuable species with a great potential for aquaculture. Bacteria play an important role in regulating the health of the early life stages. The present article is the first broad-range molecular analysis of bacterial communities in larvae of the Atlantic halibut (Hippoglossus hippoglossus). DNA was extracted from larvae, water and silo biofilm from hatcheries in Norway, Scotland, Iceland and Canada. Eubacterial 16S rRNA gene fragments were amplified by polymerase chain reaction (PCR) with broad-range primers. Sequences spanning the hyper variable V3 region representing individual bacterial species were separated into community profiles by denaturing gradient gel electrophoresis (DGGE). The profiles revealed simple communities after hatching and bacterial succession following growth. Sequencing and phylogenetic analysis of excised DGGE bands suggested aerobic heterotrophs related to groups of Pseudomonas, Janthinobacterium and possibly Marinomonas to be the primary colonisers of the larvae. After onset of feeding, fermentative species (Vibrio) were detected as well. Comparative analysis of bacterial communities from different geographical regions indicated that larvae of the Atlantic halibut possess a distinct and specific normal flora. [Copyright &y& Elsevier]

Staphylococcus aureus cell envelope antigen is a new candidate for the induction of IgA nephropathy. Background. IgA nephropathy is the most common form of glomerulonephritis worldwide. We previously reported a novel form of glomerulonephritis with glomerular IgA deposits following methicillin-resistant Staphylococcus aureus ( S. aureus) infection. We investigated the role of S. aureus related antigens in the immunopathogenesis of IgA nephropathy by producing several monoclonal antibodies against S. aureus surface antigens and determining the epitopes of deposited antigens in patients with IgA nephropathy. Methods. Cell membrane proteins were isolated from cultured S. aureus. Mouse monoclonal antibodies against these proteins were generated, and their target epitopes were determined by antibody affinity chromatography and amino acid sequence analysis, and by monoclonal antibody screening of Escherichia coli clones transfected with plasmids from the Lambda S. aureus Genomic Library. Renal biopsy specimens from 116 patients with IgA nephropathy and 122 patients with other forms of renal disease were examined for glomerular antigen depositions by immunofluorescence microscopy. Results. The major antigen recognized by monoclonal antibodies against S. aureus cell membrane was identified as the S. aureus cell envelope antigen designated ‘probable adhesin’ (ACCESSION AP003131-77, Protein ID; BAB41819.1). In 68.1% (79/116) of renal biopsy specimens from patients with IgA nephropathy, S. aureus cell envelope antigen was localized in the glomeruli, and the data confirmed that S. aureus cell envelope antigen was co-localized with IgA antibody in the glomeruli. No deposition of this antigen was detected in the glomeruli of patients with non-immune complex deposit forms of glomerulonephritis. Conclusion. S. aureus cell envelope antigen is a new candidate for the induction of IgA nephropathy. [ABSTRACT FROM AUTHOR]