Bordetella Bronchiseptica Research Articles

PEGylated nano-Rehmannia glutinosa polysaccharide induces potent adaptive immunity against Bordetella bronchiseptica

Vaccines, in many cases, stimulate only too weak immunogenicity to prevent infection. Therefore, adjuvants are required during their preparation to boost the immune response. We herein developed a PEGylated nano-adjuvant based on Rehmannia glutinosa polysaccharide (RGP). The addition of PEG layer exhibits enhanced immune performance of the nano-RGP. Stimulation of dendritic cells (DCs) with PEGylated nano-RGP (pRL) led to increased proliferation and cytokine production (IL-6, IL-12, IL-1β and TNF-α). The pRL was internalized into DCs via a rapid and efficient method. The mice immunized with pRL exhibited enhanced antigen-specific serum IgG and Th1-(IFN-γ), Th2-(IL-4), and Th17-(IL-17, IL-6) cytokine production, contributing to a good anti-infection performance. Furthermore, the pRL could effectively deliver the antigen to the lymph nodes (LNs), activate DC in the LN and produce enhanced CD4+and CD8+ T-cells-derived memory (CD44high CD62Lhigh), and effector (CD44high CD62Llow) as well as functional phenotypes. Our results revealed that pRL can act as a promising adjuvant with targeted delivery of antigen due to its effective activation and robust adaptive immunity induction of DCs.

PSVI-24 Immunological Effects of a Purified 1,3/1,6 β-glucan Supplementation in Retorted Canine Diets

Abstract Yeast cell wall products are common functional ingredients capable of “priming” the immune system, specifically in reference to vaccine efficacy. Twenty-four adult, female Beagles were used in a completely randomized design. Three retorted diets were used: control diet (CON), CON plus β-glucan top-dressed daily upon time of feeding (C+B), and CON plus retorted β-glucan included in diet formulation (BG). Following a 7 d adaptation to CON, dogs were fed their respective treatment diets for 42 d and were challenged with an oral Bordetella bronchiseptica vaccine on d 14 with blood collections on d 0, 21, 28, and 42. The objectives of the present study were 1) to evaluate the effects of dietary inclusion of a yeast β-1,3/1,6 glucan (150 ppm) on apparent total tract digestibility (ATTD) of macronutrients, fecal microbiota, and peripheral blood mononuclear cells (PBMC; T-cells, B-cells, and monocytes) of adult dogs and, 2) to test the effects of retorting on the efficacy of these β-glucans. All diets were well-accepted by all dogs. ATTD of both dry matter and crude protein were greater (P < 0.05) for BG than CON and with greater (P < 0.05) energy digestibility for BG than both B+G and CON. Additionally, fecal short-chain and branched-chain fatty acids, ammonia, indole and phenol concentrations did not differ among treatments. No significant treatment by time interactions among treatment groups were observed for any analyzed PBMC. These data suggest that a 150 ppm inclusion of this yeast-derived β-glucan had no detrimental effects on ATTD, fecal characteristics and metabolites, nor any analyzed PBMC; however, due to the absence of differences in immune parameters among treatments, the effect of retorting on the efficacy of this product cannot confidently be assessed with these findings. Higher doses of yeast-derived β-glucan might be needed to elicit an immunological modulation in healthy adult dogs.

Disrupting Bordetella Immunosuppression Reveals a Role for Eosinophils in Coordinating the Adaptive Immune Response in the Respiratory Tract.

Recent findings revealed pivotal roles for eosinophils in protection against parasitic and viral infections, as well as modulation of adaptive immune responses in the gastric mucosa. However, the known effects of eosinophils within the respiratory tract remain predominantly pathological, associated with allergy and asthma. Simulating natural respiratory infections in mice, we examined how efficient and well-adapted pathogens can block eosinophil functions that contribute to the immune response. Bordetella bronchiseptica, a natural pathogen of the mouse, uses the sigma factor btrS to regulate expression of mechanisms that interfere with eosinophil recruitment and function. When btrS is disrupted, immunomodulators are dysregulated, and eosinophils are recruited to the lungs, suggesting they may contribute to much more efficient generation of adaptive immunity induced by this mutant. Eosinophil-deficient mice failed to produce pro-inflammatory cytokines, to recruit lymphocytes, to organize lymphoid aggregates that resemble Bronchus Associated Lymphoid Tissue (BALT), to generate an effective antibody response, and to clear bacterial infection from the respiratory tract. Importantly, the failure of eosinophil-deficient mice to produce these lymphoid aggregates indicates that eosinophils can mediate the generation of an effective lymphoid response in the lungs. These data demonstrate that efficient respiratory pathogens can block eosinophil recruitment, to inhibit the generation of robust adaptive immune responses. They also suggest that some post-infection sequelae involving eosinophils, such as allergy and asthma, might be a consequence of bacterial mechanisms that manipulate their accumulation and/or function within the respiratory tract.

Counter-Selection Method for Markerless Allelic Exchange in Bordetella bronchiseptica Based on sacB Gene From Bacillus subtilis.

Bordetella bronchiseptica is a gram-negative bacterium that causes respiratory tract infections. It is a natural pathogen of a wide variety of mammals, including some used as laboratory models. This makes B. bronchiseptica an ideal organism to study pathogen-host interactions in order to unveil molecular mechanisms behind pathogenic processes. Even though genetic engineering is an essential tool in this area, there are just a few reports about genome manipulation techniques in this organism. In this article we describe an allelic exchange protocol based on double crossover recombination facilitated by the Bacillus subtilis sacB gene that can be applied for partial or complete gene knockouts, single-nucleotide mutations, or even introduction of coding sequences for transcriptional fusions. In contrast to previously employed techniques, this protocol renders genetically manipulated chromosomes without foreign DNA and enables the construction of successive genome manipulation using the same vector backbone. The entire procedure has been developed for fast and reliable manipulations with a total duration of 2 weeks. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Setting up strains Basic Protocol 2: Homologous recombination (first crossing-over) Alternate Protocol: B. bronchiseptica electroporation Basic Protocol 3: Screening for sucrose-sensitive clones Basic Protocol 4: Homologous recombination (second crossing-over) Basic Protocol 5: PCR screening of putative marker-exchange mutants Support Protocol: Electrocompetent cell preparation.

Comparative Proteomic Analysis Reveals Complex Responses to Bordetella bronchiseptica Infections in the Spleen of Rabbits.

Bordetella bronchiseptica (B. bronchiseptica) causes a respiratory disease in rabbits. To determine the proteins of B. bronchiseptica in rabbits related to the disease, differentially accumulated proteins in B. bronchiseptica-infected cells are identified by comparative proteomic analysis. Comparative proteomic analysis detects 5814 proteins and quantifies 4854 of these. Fifty eight upregulated and 38 downregulated proteins are identified in spleen tissue after B. bronchiseptica infection of rabbits (both p < 0.05). The significantly enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways are ribosome, biosynthesis of amino acids, biosynthesis of amino acids, protein export, and carbon metabolism etc. (all p < 0.01). Significantly enriched KEGG pathways include 'ocu03010 ribosome' (a); 'ocu00260 glycine, serine threonine metabolism'. Analyses of control and infected spleen cells detect responses to B. bronchiseptica infection. Many differentially affected proteins are evident, and reflect different biological changes and diverse subcellular localizations between control and infected spleen cells. Infection markedly alters the expressions of proteins linked to the serine protease system, with the 'phagosome,' 'biosynthesis of amino acids,' 'glycine, serine threonine metabolism,' 'intestinal immune network for IgA production', and 'amino sugar and nucleotide sugar metabolism' associated with B. bronchiseptica infection. The result will inform studies of responses to B. bronchiseptica infections in rabbits.

A retrospective study on the presence of selected infectious agents in lung samples of cats with pneumonia.

The causative role of some infectious agents found in cases of feline pneumonia is under debate, because they are also part of the physiological microbiota of the respiratory tract of healthy animals. In this retrospective study, archived formalin-fixed and paraffin-wax-embedded lung samples of 69 severe and lethal cases of pneumonia in cats were examined by immunohistochemistry (IHC) for the detection of nine selected infectious agents: Pasteurella multocida, Bordetella bronchiseptica, Mycoplasma felis, M. gateae, Chlamydia felis, feline herpesvirus type 1, feline coronavirus, canine distemper virus, and Toxoplasma gondii. The intention was to elucidate their immediate involvement in pneumonia formation. Due to the cross-reactivity of the applied antibodies, a species-specific polymerase chain reaction (PCR) for both targeted Mycoplasma species was applied additionally. In the 42 cases (60.9%) positive for at least one pathogen, several agents were present in a high proportion of the samples (P. multocida - 34.8%, B. bronchiseptica - 29.0%), while others were present in a moderate (feline herpesvirus type 1 - 18.8%, M. gateae - 13.0%, M. felis - 10.1%) or low percentage (T. gondii - 1.4%). All samples were negative for C. felis, feline coronavirus and canine distemper virus. Mixed infections of up to four pathogens were more frequent than single infections. Mycoplasma preferably colonised lung tissue damaged by other pathogens because they never occurred as single infections. Pasteurella multocida, B. bronchiseptica, M. felis, feline herpesvirus type 1 and T. gondii showed abundant replication within lung lesions, thus suggesting a prominent role in pneumonia formation.

Heat shock enhances outer-membrane vesicle release in Bordetella spp.

Pertussis, also known as whooping cough, is caused by the Gram-negative bacterium Bordetella pertussis, an obligate human pathogen. Despite high vaccination rates in high-income countries, resurgence of pertussis cases is an occurring problem that urges the necessity of developing an improved vaccine. Likewise, the efficacy of vaccines for Bordetella bronchiseptica, which causes similar disease in pigs and companion animals, is debatable. A promising approach for novel vaccines is the use of outer membrane vesicles (OMVs). However, spontaneous OMV (sOMV) release by Bordetella spp. is too low for cost-effective vaccine production. Therefore, we investigated the influence of growth in various media commonly used for culturing Bordetella in the Bvg+, i.e. virulent, phase and of a heat shock applied to inactivate the cells on OMV production. Inactivation of the bacterial cells at 56 °C before OMV isolation greatly enhanced OMV release in both Bordetella spp. without causing significant cell lysis. The growth medium used barely affected the efficiency of OMV release but did affect the protein pattern of the OMVs. Differences were found to be related, at least in part, to different availability of the nutrient metals iron and zinc in the media and include expression of potentially relevant vaccine antigens, such as the receptors FauA and ZnuD. The protein content of OMVs released by heat shock was comparable to that of sOMVs as determined by SDS-PAGE and Western blot analysis, and their heat-modifiable electrophoretic mobility suggests that also protein conformation is unaffected. However, significant differences were noticed between the protein content of OMVs and that of a purified outer membrane fraction, with two major outer membrane proteins, porin OmpP and the peptidoglycan-associated RmpM, being underrepresented in the OMVs. Altogether, these results indicate that the application of a heat shock is potentially an important step in the development of cost-effective, OMV-based vaccines for both Bordetella spp.

Prospecting potential links between PRRSV infection susceptibility of alveolar macrophages and other respiratory infectious agents present in conventionally reared pigs

Porcine Reproductive and Respiratory Syndrome virus (PRRSV) is one of the main component of the porcine respiratory disease complex (PRDC), which strongly impact the pig production. Although PRRSV is often considered as a primary infection that eases subsequent respiratory coinfections, the possibility that other PRDC components may facilitate PRRSV infection has been largely overlooked. The main cellular targets of PRRSV are respiratory macrophages among them alveolar macrophages (AM) and pulmonary intravascular macrophages (PIM). AM, contrarily to PIM, are directly exposed to the external respiratory environment, among them co-infectious agents. In order to explore the possibility of a co-infections impact on the capacity of respiratory macrophages to replicate PRRSV, we proceed to in vitro infection of AM and PIM sampled from animals presenting different sanitary status, and tested the presence in the respiratory tract of these animals of the most common porcine respiratory pathogens (PCV2, Actinobacillus pleuropneumoniae, Mycoplasma hyopneumoniae, Mycoplasma hyorhinis, Mycoplasma floculare, Pasteurella multocida, Bordetella bronchiseptica, Streptoccocus suis). In this exploratory study with a limited number of animals, no statistic differences were observed between AM and PIM susceptibility to in vitro PRRSV infection, nor between AM coming from animals presenting very contrasting respiratory coinfection loads.

Clinical response to 2 protocols of aerosolized gentamicin in 46 dogs with Bordetella bronchiseptica infection (2012-2018).

Background Bordetella bronchiseptica (Bb) infection commonly causes respiratory disease in dogs. Gentamicin delivered by aerosol maximizes local drug delivery without systemic absorption but clinical response to protocols remains undetermined.ObjectivesTo compare the clinical response to 2 protocols of aerosolized delivery of gentamicin in bordetellosis.AnimalsForty‐six dogs with Bb infection confirmed by culture or quantitative polymerase chain reaction on bronchoalveolar lavage.MethodsRetrospective study. Administration of aerosolized gentamicin for ≥10 minutes q12h for ≥3 weeks using 4 mg/kg diluted with saline (group 1) or undiluted 5% solution (group 2). Clinical response firstly assessed after 3‐4 weeks and treatment pursued by 3‐weeks increments if cure not reached. Cure defined as absence of cough persisting at least a week after treatment interruption.ResultsDemographic data were similar between both groups. Clinical cure at 3‐4 weeks was more frequently observed with the use of undiluted solution (19/33 vs 3/13 dogs, P = .03) in association with a shorter median duration of treatment (4 vs 6 weeks, P = .01). Dogs from group 2 having less than 1000 cells/μL in lavage were also more likely to be cured at 3‐4 weeks than dogs with more than 1000 cells/μL [9/9 vs 10/19, P = .006] and median duration of treatment in that subgroup of animals was reduced (3 vs 5 weeks, P = .02).Conclusion and Clinical ImportanceAerosolized delivery of gentamicin seems effective for inducing clinical cure in Bb infection. Clinical response appears better using undiluted 5% solution, particularly in the subgroup of dogs having less than 1000 cells/μL in lavage.

Cytotoxicity of the effector protein BteA was attenuated inBordetella pertussisby insertion of an alanine residue.

Bordetella bronchiseptica and Bordetella pertussis are closely related respiratory pathogens that evolved from a common bacterial ancestor. While B. bronchiseptica has an environmental reservoir and mostly establishes chronic infections in a broad range of mammals, B. pertussis is a human-specific pathogen causing acute pulmonary pertussis in infants and whooping cough illness in older humans. Both species employ a type III secretion system (T3SS) to inject a cytotoxic BteA effector protein into host cells. However, compared to the high BteA-mediated cytotoxicity of B. bronchiseptica, the cytotoxicity induced by B. pertussis BteA (Bp BteA) appears to be quite low and this has been attributed to the reduced T3SS gene expression in B. pertussis. We show that the presence of an alanine residue inserted at position 503 (A503) of Bp BteA accounts for its strongly attenuated cytotoxic potency. The deletion of A503 from Bp BteA greatly enhanced the cytotoxic activity of B. pertussis B1917 on mammalian HeLa cells and expression of Bp BteAΔA503 was highly toxic to Saccharomyces cerevisiae cells. Vice versa, insertion of A503 into B. bronchiseptica BteA (Bb BteA) strongly decreased its cytotoxicity to yeast and HeLa cells. Moreover, the production of Bp BteAΔA503 increased virulence of B. pertussis B1917 in the mouse model of intranasal infection (reduced LD50) but yielded less inflammatory pathology in infected mouse lungs at sublethal infectious doses. This suggests that A503 insertion in the T3SS effector Bp BteA may represent an evolutionary adaptation that fine-tunes B. pertussis virulence and host immune response.

Antimicrobial Susceptibility Pattern of Porcine Respiratory Bacteria in Spain.

The monitoring of antimicrobial susceptibility of pig pathogens is critical to optimize antimicrobial treatments and prevent development of resistance with a one-health approach. The aim of this study was to investigate the antimicrobial susceptibility patterns of swine respiratory pathogens in Spain from 2017 to 2019. Bacterial isolation and identification were carried out following standardized methods from samples coming from sacrificed or recently deceased pigs with acute clinical signs compatible with respiratory tract infections. Minimum inhibitory concentration (MIC) values were determined using the broth microdilution method containing a total of 10 and 7–8 antimicrobials/concentrations respectively, in accordance with the recommendations presented by the Clinical and Laboratory Standards Institute (CLSI). The obtained antimicrobial susceptibility varies between pig respiratory pathogens. Actinobacillus pleuropneumoniae (APP) and Pasteurella multocida (PM) were highly susceptible (≥90%) to ceftiofur, florfenicol and macrolides (tilmicosin, tildipirosin and tulathromycin). However, the antimicrobial susceptibility was intermediate (>60% but <90%) for amoxicillin and enrofloxacin in the case of APP and sulfamethoxazole/trimethropim and tiamulin in the case of PM. Both bacteria showed low (<60%) antimicrobial susceptibility to doxycycline. Finally, Bordetella bronchiseptica was highly susceptible only to tildipirosin and tulathromycin (100%) and its susceptibility for florfenicol was close to 50% and <30% for the rest of the antimicrobial families tested. These results emphasize the need of determining antimicrobial susceptibility in pig respiratory cases in order to optimize the antimicrobial treatment in a case-by-case scenario.

Antibiotic sensitivity of common respiratory bacteria of pig from Hubei province, China

The use of antimicrobials for feeding and treatment is crucial to animal health. However, continuous use of antibiotics is contributing to emergence and widespread of antibiotic resistance. This study aimed to investigate the antimicrobial resistance of five major respiratory pathogens in pigs of Hubei province, China, from October to December, 2019. Antibiotic susceptibility testing for Streptococcus suis, Haemophilus parasuis, Pasteurella multocida, Bordetella bronchiseptica and Actinobacillus pleuropneumoniae was determined to representatives of relevant antibiotic classes. Streptococcus suis isolates were mostly sensitive to beta-lactams, whereas high levels of resistance were observed to quinolones, gentamycin, doxycycline, trimethoprime and lincomycin. For H. parasuis, P. multocida and A. pleuropneumoniae of Pasteurellaceae family, the susceptibility to beta-lactams and quinolones was displayed. Most B. bronchiseptica isolates were sensitive to doxycycline azithomycin, polymycin whereas high resistance levels to betalactams, aminoglycosides and quinolones were recorded. This study obtained practical data for later studies and usage to combat infections due to respiratory bacteria.

Occurrence and Predictors of Bacterial Respiratory Tract Infections and Antimicrobial Resistance Among Isolates From Dogs Presented With Lower Respiratory Tract Infections at a Referral Veterinary Hospital in South Africa.

Background: Respiratory tract infections (RTIs) associated with Pasteurella multocida, Bordetella bronchiseptica, Streptococci, Staphylococci, and Pseudomonas species have been reported in dogs. The objective of this study was to investigate the occurrence and predictors of bacterial RTIs and antimicrobial resistance among samples from dogs with lower RTIs at a referral veterinary teaching hospital in South Africa.Methods: Records of 157 dogs with lower RTIs presented to the veterinary teaching hospital between 2007 and 2013 were included in the study. Crude and factor-specific proportions of RTIs and antimicrobial resistance by breed, season, year, sex, age category, and specimen type were computed. Chi-square or Fisher's exact tests were used to compare proportions of RTIs and antimicrobial resistant isolates across categorical variables. Associations between breed, season, year, sex, age, specimen, and odds of RTIs or multidrug resistance were assessed using Generalized Estimating Equations.Results: There was only one sample per clinical case and bacterial RTIs were observed in 53.5% of the samples tested. Pasteurella species (23.5%) were more common than other species. Almost all (99.5%) isolates were resistant to at least one antimicrobial, while 64.7% were multidrug resistant (MDR). Additionally, 17.0% and 3.3% showed evidence of extensive drug resistance (XDR) and pan-drug resistance (PDR), respectively. The majority of MDR isolates were resistant to penicillin-G (90.9%), lincomycin (100%), tylosine (75.8%), lincospectin (73.7%), ampicillin (72.5%), and kanamycin (68.4%). None of the investigated predictors had significant association with RTIs or antimicrobial resistance.Conclusion: Pasturella species were the most common causes of RTIs. The high levels of MDR and the presence of both XDR and PDR isolates raise the question of the effectiveness of the current antimicrobial therapy used in patients with RTIs in referral hospitals. Given the high level of resistance observed in this study, it is advisable that the choice of antimicrobials for treatment of RTIs be based on antibiograms. This will ensure use of the most efficacious antimicrobials and will minimize treatment failures among cases presented with RTIs.

Investigation of the effects of an ultraviolet germicidal irradiation system on concentrations of aerosolized surrogates for common veterinary pathogens.

To determine whether exposure to UV germicidal irradiation (UVGI) reduces concentrations of viable aerosolized microorganisms (attenuated strains of common veterinary pathogens) in a simulated heating, ventilation, and air conditioning (HVAC) system. 42 air samples seeded with bacteriophage MS2 or attenuated strains of Bordetella bronchiseptica, feline calicivirus, feline herpesvirus-1, canine parvovirus, or canine distemper virus (6/microorganism) or with no microorganisms added (6). A simulated HVAC unit was built that included a nebulizer to aerosolize microorganisms suspended in phosphate-buffered water, a fan to produce airflow, 2 UVGI bulb systems, and an impinger for air sampling. Ten-minute trials (3 with UVGI, 3 without UVGI, and 1 negative control) were conducted for each microorganism. Impingers collected microorganisms into phosphate-buffered water for subsequent quantification with culture-based assays. Results for samples yielding no target microorganisms were recorded as the assay's lower limit of detection. Statistical analysis was not performed. The UVGI treatment resulted in subjectively lower concentrations of viable MS2, B bronchiseptica, and canine distemper virus (arithmetic mean ± SD log10 microorganism reduction, 2.57 ± 0.47, ≥ 3.45 ± 0.24, and ≥ 1.50 ± 0.25, respectively) collected from air. Feline herpesvirus-1 was detected in only 1 sample without and no samples with UVGI treatment. Feline calicivirus and canine parvovirus were not detectable in any collected samples. Results for some surrogates of veterinary pathogens suggested a potential benefit to supplementing manual disinfection practices with UVGI-based air cleaning systems in animal care environments. Further research is needed to investigate the utility of UVGI in operating HVAC systems.

Structure, Function, Regulation and Phylogenetic Relationship of ZIP Family Transporters of Plants.

Zinc (Zn) is an essential micronutrient for plants and humans. Nearly 50% of the agriculture soils of world are Zn-deficient. The low availability of Zn reduces the yield and quality of the crops. The zinc-regulated, iron-regulated transporter-like proteins (ZIP) family and iron-regulated transporters (IRTs) are involved in cellular uptake of Zn, its intracellular trafficking and detoxification in plants. In addition to Zn, ZIP family transporters also transport other divalent metal cations (such as Cd2+, Fe2+, and Cu2+). ZIP transporters play a crucial role in biofortification of grains with Zn. Only a very limited information is available on structural features and mechanism of Zn transport of plant ZIP family transporters. In this article, we present a detailed account on structure, function, regulations and phylogenetic relationships of plant ZIP transporters. We give an insight to structure of plant ZIPs through homology modeling and multiple sequence alignment with Bordetella bronchiseptica ZIP (BbZIP) protein whose crystal structure has been solved recently. We also provide details on ZIP transporter genes identified and characterized in rice and other plants till date. Functional characterization of plant ZIP transporters will help for the better crop yield and human health in future.

Green Synthesis of MnO Nanoparticles Using Abutilon indicum Leaf Extract for Biological, Photocatalytic, and Adsorption Activities.

We report the synthesis of MnO nanoparticles (AI-MnO NAPs) using biological molecules of Abutilon indicum leaf extract. Further, they were evaluated for antibacterial and cytotoxicity activity against different pathogenic microbes (Escherichia coli, Bordetella bronchiseptica, Staphylococcus aureus, and Bacillus subtilis) and HeLa cancerous cells. Synthesized NAPs were also investigated for photocatalytic dye degradation potential against methylene blue (MB), and adsorption activity against Cr(VI) was also determined. Results from Scanning electron microscope (SEM), X-ray powder diffraction (XRD), Energy-dispersive X-ray (EDX), and Fourier-transform infrared spectroscopy (FTIR) confirmed the successful synthesis of NAPs with spherical morphology and crystalline nature. Biological activity results demonstrated that synthesized AI-MnO NAPs exhibited significant antibacterial and cytotoxicity propensities against pathogenic microbes and cancerous cells, respectively, compared with plant extract. Moreover, synthesized AI-MnO NAPs demonstrated the comparable biological activities results to standard drugs. These excellent biological activities results are attributed to the existence of the plant’s biological molecules on their surfaces and small particle size (synergetic effect). Synthesized NAPs displayed better MB-photocatalyzing properties under sunlight than an ultraviolet lamp. The Cr(VI) adsorption result showed that synthesized NAPs efficiently adsorbed more Cr(VI) at higher acidic pH than at basic pH. Hence, the current findings suggest that Abutilon indicum is a valuable source for tailoring the potential of NAPs toward various enhanced biological, photocatalytic, and adsorption activities. Consequently, the plant’s biological molecule-mediated synthesized AI-MnO NAPs could be excellent contenders for future therapeutic applications.

Antimicrobial resistance in bacteria isolated from pigs with respiratory clinical signs in Brazil

Antimicrobial resistance is a current and important issue to public health, and it is usually associated with the indiscriminate use of antimicrobials in animal production. This study aimed to evaluate the antimicrobial susceptibility profile in bacterial isolates from pigs with clinical respiratory signs in Brazil. One hundred sixty bacterial strains isolated from pigs from 51 pig farms in Brazil were studied. In vitro disk-diffusion method was employed using 14 antimicrobial agents: amoxicillin, penicillin, ceftiofur, ciprofloxacin, enrofloxacin, chlortetracycline, doxycycline, oxytetracycline, tetracycline, erythromycin, tilmicosin, florfenicol, lincomycin, and sulfadiazine/trimethoprim. The majority of isolates were resistant to at least one antimicrobial agent (98.75%; 158/160), while 31.25% (50/160) of the strains were multidrug resistant. Streptococcus suis and Bordetella bronchiseptica were the pathogens that showed higher resistance levels. Haemophilus parasuis showed high resistance levels to sulfadiazine/trimethoprim (9/18=50%). We observed that isolates from the midwestern and southern regions exhibited four times greater chance of being multidrug resistant than the isolates from the southeastern region studied. Overall, the results of the present study showed a great level of resistance to lincomycin, erythromycin, sulfadiazine/trimethoprim, and tetracycline among bacterial respiratory pathogens isolated from pigs in Brazil. The high levels of antimicrobial resistance in swine respiratory bacterial pathogens highlight the need for the proper use of antimicrobials in Brazilian pig farms.

Genomics of Serratia marcescens Isolates Causing Outbreaks in the Same Pediatric Unit 47 Years Apart: Position in an Updated Phylogeny of the Species.

The first documented nosocomial outbreak caused by Serratia marcescens in Spain occurred in 1969 at the neonatal intensive care unit (NICU) of the tertiary La Paz Children’s Hospital in Madrid, Spain, and based on the available phenotyping techniques at this time, it was considered as a monoclonal outbreak. Only 47 years later, another S. marcescens outbreak of an equivalent dimension occurred at the same NICU. The aim of the present study was to study isolates from these historical and contemporary outbreaks by phenotypic analysis and whole-genome sequencing techniques and to position these strains along with 444 publicly available S. marcescens genomes, separately comparing core genome and accessory genome contents. Clades inferred by both approaches showed high correlation, indicating that core and accessory genomes seem to evolve in the same manner for S. marcescens. Nine S. marcescens clusters were identified, and isolates were grouped in two of them according to sampling year. One exception was isolate 13F-69, the most genetically distant strain, located in a different cluster. Categorical functions in the annotated accessory genes of both collections were preserved among all isolates. No significant differences in frequency of insertion sequences in historical (0.18–0.20)—excluding the outlier strain—versus contemporary isolates (0.11–0.19) were found despite the expected resting effect. The most dissimilar isolate, 13F-69, contains a highly preserved plasmid previously described in Bordetella bronchiseptica. This strain exhibited a few antibiotic resistance genes not resulting in a resistant phenotype, suggesting the value of gene down expression in adaptation to long-term starvation.

Analysis of the lung microbiota in dogs with Bordetella bronchiseptica infection and correlation with culture and quantitative polymerase chain reaction

Infection with Bordetella bronchiseptica (Bb), a pathogen involved in canine infectious respiratory disease complex, can be confirmed using culture or qPCR. Studies about the canine lung microbiota (LM) are recent, sparse, and only one paper has been published in canine lung infection. In this study, we aimed to compare the LM between Bb infected and healthy dogs, and to correlate sequencing with culture and qPCR results. Twenty Bb infected dogs diagnosed either by qPCR and/or culture and 4 healthy dogs were included. qPCR for Mycoplasma cynos (Mc) were also available in 18 diseased and all healthy dogs. Sequencing results, obtained from bronchoalveolar lavage fluid after DNA extraction, PCR targeting the V1–V3 region of the 16S rDNA and sequencing, showed the presence of Bb in all diseased dogs, about half being co-infected with Mc. In diseased compared with healthy dogs, the β-diversity changed (P = 0.0024); bacterial richness and α-diversity were lower (P = 0.012 and 0.0061), and bacterial load higher (P = 0.004). Bb qPCR classes and culture results correlated with the abundance of Bb (r = 0.71, P < 0.001 and r = 0.70, P = 0.0022). Mc qPCR classes also correlated with the abundance of Mc (r = 0.73, P < 0.001). Bb infection induced lung dysbiosis, characterized by high bacterial load, low richness and diversity and increased abundance of Bb, compared with healthy dogs. Sequencing results highly correlate with qPCR and culture results showing that sequencing can be reliable to identify microorganisms involved in lung infectious diseases.

Characterization of a Unique Bordetella bronchiseptica vB_BbrP_BB8 Bacteriophage and Its Application as an Antibacterial Agent

Bordetella bronchiseptica, an emerging zoonotic pathogen, infects a broad range of mammalian hosts. B. bronchiseptica-associated atrophic rhinitis incurs substantial losses to the pig breeding industry. The true burden of human disease caused by B. bronchiseptica is unknown, but it has been postulated that some hypervirulent B. bronchiseptica isolates may be responsible for undiagnosed respiratory infections in humans. B. bronchiseptica was shown to acquire antibiotic resistance genes from other bacterial genera, especially Escherichia coli. Here, we present a new B. bronchiseptica lytic bacteriophage—vB_BbrP_BB8—of the Podoviridae family, which offers a safe alternative to antibiotic treatment of B. bronchiseptica infections. We explored the phage at the level of genome, physiology, morphology, and infection kinetics. Its therapeutic potential was investigated in biofilms and in an in vivo Galleria mellonella model, both of which mimic the natural environment of infection. The BB8 is a unique phage with a genome structure resembling that of T7-like phages. Its latent period is 75 ± 5 min and its burst size is 88 ± 10 phages. The BB8 infection causes complete lysis of B. bronchiseptica cultures irrespective of the MOI used. The phage efficiently removes bacterial biofilm and prevents the lethality induced by B. bronchiseptica in G. mellonella honeycomb moth larvae.