Low-Level Antibiotic Resistance among Staphylococcus aureus and Gram-Negative Pathogens from Infected Skin and Soft Tissues in Rural Kenya

Introduction: Bacterial skin and soft tissue infections (SSTIs) are a cause of frequent inpatient and outpatient care visits whose causative agents are associated with a high antimicrobial resistance burden. For insights on antimicrobial susceptibilities in a rural setting, we examined specimens from suspected SSTIs from two public health facilities in Kenya. We additionally assessed antibiotic use, appropriateness of empiric therapy and risk factors for SSTI. Methodology: Between 2021 and 2023, 265 patients at Kisii and Nyamira County Referral hospitals were enrolled. Wound swabs/aspirates were collected and processed following standard microbiological procedures. Identification and antimicrobial susceptibility were performed using the VITEK 2 Compact platform. Demographic, clinical, and microbiological data were analyzed with R Statistical software. Results: S. aureus was isolated in 16.2% (43/265) of patients with a methicillin resistance (MRSA) proportion of 14% (6/43). While 13/15 drugs elicited susceptibilities ranging from 84% - 100%, penicillin (16%) and trimethoprim-sulfamethoxazole [TMP-SXT] (23%) yielded the lowest susceptibilities. Escherichia coli (n = 33), Klebsiella pneumoniae (n = 8), Pseudomonas aeruginosa (n = 8), and Citrobacter species (n = 4) were the most commonly isolated gram-negative species. Gram-negative strains showed high susceptibilities to most of the tested drugs (71% - 100%) with the exception of ampicillin (18%), TMP-SXT (33%), and first and second generation cephalosporins. Conclusions: The low MRSA prevalence and generally high antibiotic susceptibilities for S. aureus and gram-negative bacteria present opportunities for antibiotic stewardship in the study setting. Diminished susceptibilities against penicillin/ampicillin and TMP-SXT accord with prevailing local data and add a layer of evidence for their cautious empiric use.

Immunogenomics for identification of disease resistance genes in pigs: a review focusing on Gram-negative bacilli

Over the past years, infectious disease has caused enormous economic loss in pig industry. Among the pathogens, gram negative bacteria not only cause inflammation, but also cause different diseases and make the pigs more susceptible to virus infection. Vaccination, medication and elimination of sick pigs are major strategies of controlling disease. Genetic methods, such as selection of disease resistance in the pig, have not been widely used. Recently, the completion of the porcine whole genome sequencing has provided powerful tools to identify the genome regions that harboring genes controlling disease or immunity. Immunogenornics, which combines DNA variations, transcriptorne, immune response, and QTL mapping data to illustrate the interactions between pathogen and host immune system, will be an effective genomics tool for identification of disease resistance genes in pigs. These genes will be potential targets for disease resistance in breeding programs. This paper reviewed the progress of disease resistance study in the pig focusing on Gram-negative bacilli. Major porcine Gram-negative bacilli and diseases, suggested candidate genes/pathways against porcine Gram-negative bacilli, and distributions of QTLs for immune capacity on pig chromosomes were summarized. Some tools for immunogenomics research were described. We conclude that integration of sequencing, whole genome associations, functional genomics studies, and immune response information is necessary to illustrate molecular mechanisms and key genes in disease resistance.

Detection of 16S rRNA Methylase Genes in Gram-Negative Bacilli Isolated from Hospitals in Changchun, China

Methylation of 16S rRNA is an important mechanism of aminoglycoside resistance among gram-negative pathogens. In this report, 16S rRNA methylase genes were amplified using PCR among gram-negative bacillus isolates from hospitals in the Changchun area of China and 16S rRNA methylase genotypes (armA, rmtB, rmtA, rmtC, rmtD, and npmA) were identified by direct sequencing. Fifty of the isolates (43.1%) harbored 16S rRNA methylase genes. The common 16S rRNA methylase genes were armA and rmtB (12.1% and 31.0%, respectively), whereas the rmtA, rmtC, rmtD, and npmA genes were absent from the sample. It suggests that the predominant 16S rRNA methylase genes among gramnegative bacilli in the Changchun area are armA and rmtB.

Detection of the Production of Klebsiella Pneumoniae Carbapenemase, New Delhi Metallo-Beta-Lactamase and Oxacillinase-48-Type Carbapenemases by Gram-Negative Bacilli in Resource-Limited Setting

Background: The increasing resistance of bacteria to various antibiotics is a worldwide public health issue. Carbapenems that have elicited great hope in treating infections caused by multidrug-resistant germs have seen their efficacy narrowed over time with the emergence of other novel resistance mechanisms, notably the production of Carbapenemases. Methods: A prospective cross-sectional study was conducted from May 2017 to May 2018 in Douala (Cameroon) to detect carbapenemase-producing Gram-negative bacilli. Isolated strains were identified using the Vitek2TM system. Antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method on agar plates with 20 selected commercially available antibiotic discs. The bacterial strains were tested for the production of three Carbapenemases (OXA-48, NDM, KPC), using an immuno-chromatographic technique, with the “RESIST-3 O.K.N. K-SeT” rapid detection kit. Results: During the study period, 1687 strains of Gram-negative bacilli were isolated in selected laboratories with a total of 200 multi-resistant strains identified (11.9%). Among the multi-resistant strains, E. coli was the species most represented in Enterobacteriaceae (27.5%) followed by K. pneumoniae (15.5%) and the non-fermenting Gram-negative bacilli were predominantly P. aeruginosa (20.5%). These strains mainly came from urine and pus, i.e. 41% and 32% respectively. Thirty-two (16%) strains produced one of the Carbapenemases with a higher frequency at the General Hospital (84%). NDM-type carbapenemase was the most frequently identified (8.5%), OXA-48 type 7.5%, and no KPC production was observed. Among the Enterobacteriaceae 22.9% produced Carbapenemases and only 5.1% of the non-fermenting bacilli produced these enzymes. The isolates strains were completely resistant to all antibiotics except Amikacin and Fosfomycin. The strains producing the NDM-type carbapenemase showed higher rates of resistance to almost all of the antibiotics tested. Conclusion: Multidrug-resistant strains are experiencing an increase in evolution. The apparition of strains producing Carbapenemases prominently, the NDM and OXA-48 favor this increase. The activities of antibiotics with high efficacies on these strains are low.

High mortality associated with gram-negative bacterial bloodstream infection in liver transplant recipients undergoing immunosuppression reduction

BACKGROUND Immunosuppression is an important factor in the incidence of infections in transplant recipient.Few studies are available on the management of immunosuppression(IS)treatment in the liver transplant(LT)recipients complicated with infection.The aim of this study is to describe our experience in the management of IS treatment during bacterial bloodstream infection(BSI)in LT recipients and assess the effect of temporary IS withdrawal on 30 d mortality of recipients presenting with severe infection.AIM To assess the effect of temporary IS withdrawal on 30 d mortality of LT recipients presenting with severe infection.METHODS A retrospective study was conducted with patients diagnosed with BSI after LT in the Department of Liver Surgery,Renji Hospital from January 1,2016 through December 31,2017.All recipients diagnosed with BSI after LT were included.Univariate and multivariate Cox regression analysis of risk factors for 30 d mortality was conducted in the LT recipients with Gram-negative bacterial(GNB)infection.RESULTS Seventy-four episodes of BSI were identified in 70 LT recipients,including 45 episodes of Gram-positive bacterial(GPB)infections in 42 patients and 29 episodes of GNB infections in 28 patients.Overall,IS reduction(at least 50%dose reduction or cessation of one or more immunosuppressive agent)was made in 28(41.2%)cases,specifically,in 5(11.9%)cases with GPB infections and 23(82.1%)cases with GNB infections.The 180 d all-cause mortality rate was 18.5%(13/70).The mortality rate in GNB group(39.3%,11/28)was significantly higher than that in GPB group(4.8%,2/42)(P=0.001).All the deaths in GNB group were attributed to worsening infection secondary to IS withdrawal,but the deaths in GPB group were all due to graft-versus-host disease.GNB group was associated with significantly higher incidence of intra-abdominal infection,IS reduction,and complete IS withdrawal than GPB group(P<0.05).Cox regression showed that rejection(adjusted hazard ratio 7.021,P=0.001)and complete IS withdrawal(adjusted hazard ratio 12.65,P=0.019)were independent risk factors for 30 d mortality in patients with GNB infections after LT.CONCLUSION IS reduction is more frequently associated with GNB infection than GPB infection in LT recipients.Complete IS withdrawal should be cautious due to increased risk of mortality in LT recipients complicated with BSI.

Association between infections caused by multidrug-resistant gram-negative bacteria and mortality in critically ill patients

The incidence of gram-negative multidrug-resistant(MDR) bacterial pathogens is increasing in hospitals and particularly in the intensive care unit(ICU) setting. The clinical consequences of infections caused by MDR pathogens remain controversial. The purpose of this review is to summarize the available data concerning the impact of these infections on mortality in ICU patients. Twenty-four studies, conducted exclusively in ICU patients, were identified through Pub Med search over the years 2000-2015. Bloodstream infection was the only infection examined in eight studies, respiratory infections in four and variable infections in others. Comparative data on the appropriateness of empirical antibiotic treatment were provided by only seven studies. In ten studies the presence of antimicrobial resistance was not associated with increased mortality; on the contrary, in other studies a significant impact of antibiotic resistance on mortality was found, though, sometimes, mediated by inappropriate antimicrobial treatment. Therefore, a direct association between infections due to gram-negative MDR bacteria and mortality in ICU patients cannot be confirmed. Sample size, presence of multiple confounders and other methodological issues may influence the results. These data support the need for further studies to elucidate the real impact of infections caused by resistant bacteria in ICU patients.

Seafood as a Reservoir of Gram-negative Bacteria Carrying Integrons and Antimicrobial Resistance Genes in Japan

PCR and DNA sequencing were used to screen and characterize integrons and resistance genes in Gram-negative bacteria isolated from seafood products in Japan.A total of 215 Gram-negative bacteria were isolated from local and imported seafood samples collected from retail markets in Hiroshima Prefecture.Class 1 integrons containing gene cassettes encoding resistance to trimethoprim

Impact of Polymyxin Resistance on Virulence and Fitness among Clinically Important Gram-Negative Bacteria

Humanity is facing an enormous and growing worldwide threat from the emergence of multi-drug-resistant(MDR)Gram-negative bacteria such as Escherichia coli,Klebsiella pneumoniae,and Acinetobacter baumannii.Polymyxin B and E(colistin)constitute the last-line therapies for treating MDR Gram-negative bacteria.Polymyxin is a cationic antibacterial peptide that can destroy the outer membrane of Gram-negative bacteria.With the increasing clinical application of polymyxin,however,there have been many reports of the occurrence of polymyxin-resistant Gram-negative bacteria.This resistance is mainly mediated by the modification or complete loss of lipopolysaccharide(LPS).LPS is also a virulence factor of Gram-negative bacteria,and alterations of LPS may correlate with virulence.Although it is generally believed that the biological costs associated with drug resistance may enable benign susceptible bacteria to overcome resistant bacteria when antibiotic pressure is reduced,some studies have shown that polymyxin-resistant bacteria are associated with higher virulence and greater fitness compared with their susceptible counterparts.To predict the development of polymyxin resis-tance and evaluate interventions for its mitigation,it is important to understand the relative biological cost of polymyxin resistance compared with susceptibility.The impact of polymyxin resistance mecha-nisms on the virulence and fitness of these three Gram-negative bacteria are summarized in this review.

An Improved Approach for Rapidly Identifying Different Types of Gram-Negative Bacterial Secreted Proteins

Protein secretion plays an important role in bacterial lifestyles. In Gram-negative bacteria, a wide range of proteins are secreted to modulate the interactions of bacteria with their environments and other bacteria via various secretion systems. These proteins are essential for the virulence of bacteria, so it is crucial to study them for the pathogenesis of diseases and the development of drugs. Using amino acid composition (AAC), position-specific scoring matrix (PSSM) and N-terminal signal peptides, two different substitution models are firstly constructed to transform protein sequences into numerical vectors. Then, based on support vector machine (SVM) and the “one to one”?algorithm, a hybrid multi-classifier named SecretP v.2.2 is proposed to rapidly and accurately?distinguish different types of Gram-negative?bacterial secreted proteins. When performed on the same test set for a comparison with other methods, SecretP v.2.2 gets the highest total sensitivity of 93.60%. A public independent dataset is used to further test the power of SecretP v.2.2 for predicting NCSPs, it also yields satisfactory results.

Metallo-β-lactamase producing nonfermentative gram-negative bacteria:An increasing clinical threat among hospitalized patients

Objective:To detect and evaluate the various methods for metallo-β-lactamases(MBL) production in Pseudomonas aeruginosa(P.aeruginosa) and Acinetobacter species.Methods:A total of 109 P.aeruginosa and 85 Acinetobacter species were screened for imipenem resistance by Kirby- Bauer disc diffusion methods.Detection of MBL production was(lone by imipenem-EDTA combined disc test,double disc synerygy test(DDST) and imipenem-EDTA MBL E test.Results: A total of 63(57.8%) strains of P.aeruginosa and 46(54.1%) strains of Acinetobacter spp.were found to be resistant to imipenem.Of the 63 imipenem resistant P.aeruginosa tested for MBL production.44(69.89;) were found to be positive and among 46 imipenem resistant Acinetobacter. 19(41.3%) were shown to be the MBL producers.Conclusions:Imipenem-EDTA combined disc test and MBL E test are equally effective for MBL detection in both P.aeruginosa and Acinetobacter spp.,but given the cost-constraints,combined disc can be used as a convenient screening method in the clinical microbiology laboratory.

Epidemiological Characteristics, Resistance Patterns and Spread of Gram-Negative Bacteria Related to Colonization of Patients in Intensive Care Units

Our aim was to determine the epidemiological characteristics, the resistance patterns and the spread of Gram negative bacteria related to colonization of patients in adult Intensive Care Units. Methods: A prospective cohort of patients colonized and/or infected with Gram negative bacteria was conducted at two adult ICUs from hospitals in Brazil (April 2012 to February 2013). Nasal, groin and perineum swabs were performed. Samples were incubated on MacConkey and cetrimide agar (48 h at 37℃) and identification tests (Vitek-BioMérieux), antibiogram (Bauer-Kirby method), Carba NP test, Polymerase Chain Reaction (PCR) and sequencing were performed. The patterns of resistant microorganisms were compared by rep-PCR (Diversilab). Results: There were 53 cases of colonization. In these cases, we identified imipenem-resistant Acinetobacter baumannii (51%), Pseudomonas aeruginosa (32%), Klebsiella pneumoniae ESBL (38%) or imipenem resistant (5.6%). The use of antimicrobials and medical devices were related to colonization (p The resistance patterns expressed by Klebsiella pneumoniae were ESBL (CTX-M, SHV e TEM) and KPC2. A verified profile of Acinetobacter baumannii was related to OXA-23 and OXA-253 (OXA-143 variant). The profiles ESBL and KPC2 expressed by Klebsiella pneumoniae were distributed between the both ICUs. The distribution of OXA-23 and OXA-253 was verified only in one ICU. The similarity of strains ranged from 80% to 95%, highlighting the horizontal transference of these microorganisms.

Prevalence and Antimicrobial Resistance of Gram-Negative Bacteria Isolates in Shellfish Samples from Two River Estuaries in South-South Nigeria

Antibiotic resistant bacteria pathogens remain the leading cause of shellfish borne diseases and a major health threat to humans worldwide. The objectives of this study were to isolate, identify, and determine the antibiotic resistance patterns of Gram-negative bacteria from shellfish. We analyzed a total of 540 shellfish (117 clams, 88 oysters, and 136 periwinkles) samples collected from different vendors at Iko and Douglas Creeks in Akwa Ibom State, South-South Nigeria. Conventional cultural techniques, morphological, biochemical characteristics, and PCR amplification were used to identify the bacterial isolates. Antibiotic susceptibility tests (Kirby-Bauer disk diffusion method) and ESBL phenotype (disk) of the isolates were performed. One hundred and thirty-five (135) Gram-negative bacteria comprising 5 genera and 14 species were detected at a prevalence of: Alcaligenes faecalis TRB-7 38 (28.2%), Pseudomonas oryzihabitans strain KCB005 16 (11.9%), Paenalcaligenes retgerii strain B5 12 (8.9%) Pseudomonas aeruginosa JB2 10 (7.4%), Providencia stuartii DMC-28b 9 (6.7%), Alcaligenes species TLT151 8 (5.9%), Pseudomonas aeruginosa CIFRI DTSB1 7 (5.2%), Paenalcaligenes species UN24 7 (5.2%), Alcaligenes faecalis BT10 7 (5.2%), Vibrio species strain PrVy108 6 (4.4%), Pseudomonas xiamenensis C10-2 5 (3.7%), Providencia vemicola Bu15_38 4 (2.9%), Pseudomonas anguillisceptica 4029 3 (2.2%), and Pseudomonas aeruginosa N15-01092 3 (2.2%). All tested isolates showed various degrees of resistance to the thirteen antimicrobials evaluated. High levels of resistance (100%) to cefepime and imipenem were expressed by all isolates except the Providencia species. For the EBSL indicators, all isolates apart from Alcaligenes species were resistant (100%) to ceftriaxone. All Vibrio species were susceptible to norfloxacin, nalidixic acid, and ceftazidime. The identification of antibiotic resistant Gram-negative bacteria (GNARB) from shellfish in this study highlights the risk of disseminated multi-drug resistance—a serious public health concern.

Comparison of the serum contents of inflammatory mediators and oxidative stress mediators between patients with gram-positive bacteria and gram-negative bacteria infection

Objective: To compare the serum contents of inflammatory mediators and oxidative stress mediators between patients with gram-positive bacteria and gram-negative bacteria infection. Methods: Patients who were diagnosed with bloodstream bacterial infection in Zigong Third People's Hospital between March 2015 and April 2017 were selected as the research subjects and divided into gram-positive group and gram-negative group according to the results of blood culture and strain identification, and serum levels of inflammatory mediators PCT, IL-1β, IL-6, sTREM-1, TNF-α, NGAL, SAA, HPT and hs-CRP as well as oxidative stress mediators MDA, AOPP, TAC, CAT and SOD were determined. Results: Serum PCT, IL-1β, IL-6, sTREM-1, TNF-α, NGAL, SAA, HPT, hs-CRP, MDA and AOPP levels of gram-negative group were greatly higher than those of gram-positive group while TAC, CAT and SOD levels were greatly lower than those of gram-positive group. Conclusion: The changes of inflammatory mediators and oxidative stress mediators in the serum of patients with gram-negative bacteria infection are more significant than those of patients with gram-positive bacteria infection.

Granulomatous prostatitis after bacille Calmette-Guérin instillation resembles prostate carcinoma:A case report and review of the literature

BACKGROUND Bacille Calmette-Guérin(BCG)instillation is recommended in patients with nonmuscle-invasive bladder cancer who have intermediate-risk and high-risk tumors.However,granulomatous prostatitis is a rare complication induced by BCG instillation,which can easily be misdiagnosed as prostate cancer.Here,we report a case of granulomatous prostatitis that resembled prostate cancer.CASE SUMMARY A 64-year-old Chinese man with bladder cancer received BCG instillation.Three days later,he stopped BCG instillation and received anti-infective therapy due to the urinary tract infection.Three months after BCG restart,he had rising total prostate-specific antigen(PSA)(9.14 ng/mL)and decreasing free PSA/total PSA(0.09).T2-weighted images of magnetic resonance imaging(MRI)showed a 28 mm×20 mm diffuse low signal abnormality in the right peripheral zone,which was markedly hyperintense on high b-value diffusion-weighted MRI and hypointense on apparent diffusion coefficient map images.Considering Prostate Imaging Reporting and Data System score of 5 and possibility of prostate cancer,a prostate biopsy was conducted.Histopathology showed typical features of granulomatous prostatitis.The nucleic acid test for tuberculosis was positive.He was finally diagnosed with BCG-induced granulomatous prostatitis.Thereafter,he stopped BCG instillation and received anti-tuberculosis treatment.During 10 mo follow-up,he had no evidence of tumor recurrence or symptoms of tuberculosis.CONCLUSION Temporarily elevated PSA and high followed by low signal abnormality on diffusion-weighted MRI are important indicators of BCG-induced granulomatous prostatitis.

Acute suppurative terminal cholangitis:Clinical characteristics of a new subtype of acute cholangitis

Background:Acute suppurative terminal cholangitis(ASTC)is rarer than acute obstructive cholangitis and is not well studied.To explore this subtype of acute cholangitis,we described our clinical experience with ASTC.Methods:We performed a retrospective review of patients with ASTC admitted to our center from September 2014 to August 2020.We analyzed their clinical characteristics,including etiology,clinical manifestations,imaging features,treatment and prognosis.Results:A total of 32 ASTC patients were included in the analysis.The majority of the patients had a history of biliary operations,and clinical manifestations were occult and atypical.The positive rate of bacterial culture was 46.9%.All the patients had typical imaging features on computed tomography and magnetic resonance imaging.Treatment with effective antibiotics was provided as soon as diagnosis was established.After treatment,most patients had a good outcome.Elevated levels of total bilirubin,aspartate aminotransferase,procalcitonin and gamma-glutamyltransferase were the characteristics of critically ill patients and were associated with relatively poor prognosis.Conclusions:Our results demonstrated that ASTC should be recognized as a new subtype of acute cholangitis,and that earlier diagnosis and more personalized treatments are needed.

Risk factors and mortality rates of carbapenem-resistant Gram-negative bacterial infections in intensive care units

Background The prevalence of hospital-acquired infections caused by carbapenem-resistant gram-negative bacteria(CRGNB)is increasing worldwide.Several risk factors have been associated with such infections.The present study aimed to identify risk factors and determine the mortality rates associated with CRGNB infections in intensive care units.Methods This retrospective case-control study was conducted at Erciyes University Hospital(Kayseri,Turkey)between January 2017 and December 2021.Demographic and laboratory data were obtained from the Infection Control Committee data and record system.Patients who had CRGNB infection 48–72 h after hospitalization were assigned to the case group,while those who were not infected with CRGNB during hospitalization formed the control group.Risk factors,comorbidity,demographic data,and mortality rates were compared between the two groups.Results Approximately 1449 patients(8.97%)were monitored during the active follow-up period;of those,1171 patients were included in this analysis.CRGNB infection developed in 14 patients(70.00%)who had CRGNB colonization at admission;in 162(78.26%)were colonized during hospitalization,whereas 515(54.56%)were not colonized.There was no significant difference in age,sex(male/female)or comorbidities.The total length of hospital stay was statistically significantly longer(P=0.001)in the case group(median:24[interquartile range:3–378]days)than the control group(median:16[interquartile range:3–135]days).The rates of colonization at admission(25.5%;vs.10.6%,P=0.001)and mortality(64.4%vs.45.8%,P=0.001)were also significantly higher in the cases than in the control group,respectively.In the univariate analysis,prolonged hospitalization,the time from intensive care unit admission to the development of infection,presence of CRGNB colonization at admission,transfer from other hospitals,previous antibiotic use,enteral nutrition,transfusion,hemodialysis,mechanical ventilation,tracheostomy,reintubation,central venous catheter,arterial catheterization,chest tube,total parenteral nutrition,nasogastric tube use,and bronchoscopy procedures were significantly associated with CRGNB infections(P<0.05).Multivariate analysis identified the total length of stay in the hospital(odds ratio[OR]=1.02;95%confidence interval[CI]:1.01 to 1.03;P=0.001),colonization(OR=2.19;95%CI:1.53 to 3.13;P=0.001),previous antibiotic use(OR=2.36;95%CI:1.53 to 3.62;P=0.001),intubation(OR=1.59;95%CI:1.14 to 2.20;P=0.006),tracheostomy(OR=1.42;95%CI:1.01 to 1.99;P=0.047),and central venous catheter use(OR=1.62;95%CI:1.20 to 2.19;P=0.002)as the most important risk factors for CRGNB infection.Conclusions Colonization,previous use of antibiotics,and invasive interventions were recognized as the most important risk factors for infections.Future research should focus on measures for the control of these parameters.

Antibacteria Activity of Peganum harmala and Haloxylon salicornicum Leaves Extracts

The use of antibiotics in humans and animals has been marked as a significant step in health due to their effectiveness in controlling and treating bacterial infections. The misuse and overuse of antibiotics have been identified as risk factors for bacterial resistance since microorganisms adapt and develop mechanisms to defend against antibiotics. According to the Centers for Disease Protection and Control (CDC), around 23,000 individuals die every year in the United States due to antibiotic resistance complications. As a result, a demand for alternative treatments has been a goal for scientists as the microbes adapt to selective pressure. The aim of this study is to test the antibacterial activity of leaf extracts of Peganum harmala and Haloxylon salicornicum on both Gram-positive and Gram-negative bacteria on various mediums. The results of the study showed that both P. harmala and H. salicornicum inhibited the bacterial growth in two different media. The results were also compared with different common antibiotics used in both human’s and animal’s fields and showed a promising outcome as alternative antibiotics.

Bacillus sp. probiotic supplementation diminish the Escherichia coli F4ac infection in susceptible weaned pigs by influencing the intestinal immune response, intestinal microbiota and blood metabolomics

Background:Probiosis is considered a potential strategy to reduce antibiotics use and prevent postweaning diarrhea(PWD).This study investigated the effect of Bacillus amyloliquefaciens DSM25840 or Bacillus subtilis DSM25841 supplementation on growth,health,immunity,intestinal functionality and microbial profile of post-weaning pigs after enterotoxigenic E.coli(ETEC)F4 challenge.Methods:Sixty-four post-weaning piglets(7748 g±643 g)were randomly al ocated to four groups:control basal diet(CO);CO+1.28×10~6 CFU/g of B.amyloliquefaciens(BAA);CO+1.28×10~6 CFU/g feed of B.subtilis(BAS);CO+1 g colistin/kg of feed(AB).At day(d)7,animals were chal enged with 10~5 CFU/m L of ETEC F4 ac O149 and then fol owed for fecal score and performance until d 21.Blood was col ected at d 6,d 12 and d 21 for immunoglobulins,at d 8 for acute phase proteins,at d 8 and d 21 for metabolomics analysis.Jejunum was sampled for morphometry,quantification of apoptosis,cel proliferation,neutral and acid mucine and Ig A secretory cel s,and microarray analysis at d 21.Jejunum and cecum contents were col ected for microbiota at d 21.Results:AB and BAS reduced the fecal score impairment compared to CO(P<0.05)at d 14.Body weight(BW),average daily weight gain(ADWG),average daily feed intake(ADFI)and gain to feed ratio(G:F)did not differ between Bacil us groups and CO.AB improved BW at d 7,d 14 and d 21,ADWG ADFI and G:F from d 0 to d 7(P<0.05).At d 8,CO had higher plasma arginine,lysine,ornithine,glycine,serine and threonine than other groups,and higher haptoglobin than AB(P<0.05).At d 21,CO had lower blood glycine,glutamine and Ig A than BAS.Morphology,cel s apoptosis and mucins did not differ.BAS and AB increased the vil us mitotic index.Transcriptome profile of BAS and AB were more similar than CO.Gene sets related to adaptive immune response were enriched in BAA,BAS and AB.CO had enriched gene set for nuclear structure and RNA processing.CO had a trend of higher Enterobacteriaceae in cecum than the other groups(P=0.06).Conclusion:Bacil us subtilis DSM25841 treatment may reduce ETEC F4ac infection in weaned piglets,decreasing diarrhea and influencing mucosal transcriptomic profile.

Enhancement of vitamin A combined vitamin D supplementation on immune response to Bacille Calmette-Guerin vaccine revaccinated in Chinese infants

Objective:To investigate whether there is an association between diameter of bacille CalmetteGuerin(BCG)sears and effect of purified protein derivative(PPD)reaction anil to determine whether vitamin A(VA)combined vitamin I)(VD)supplementation influences the immune response to BCG revueeinated in Chinese infants.Methods:A cross-section and 3-month community-randomised trial was conducted.A total of 5 629 infants at 3,6 and 12 months of age in Junan County of China were examined for BCG scar fonnation.Then,597 revuccinated infants were randomly assigned to supplementation(n=307)and control(n=290)groups.The supplementation group were daily assigned to 1 500 IU VA and 500 IU VD for 3 months.Then all infants were subjected to skin test with PPD.Results:The diameter of BCG sears was positively con-elated with diameter of skin indurations of PPD(r=0.17,P<0.05)in the 5 629 infants.The rate of positive response to PP1)was higher in the supplementation group than in the control group(96.1%versus 89.7%,P<0.05,prevalence ratio 1.07.95%CI 1.02-1.12).The prevalence ralio of PPD response for the supplementation group compared with that for the control group was 1.07(95%CI 1.01-1.13)for the males and 1.08(95%CI 1.00-1.17)for the females.For the supplementation group,the males got larger tuberculin induration than the females[(0.73±0.2l)cm versus(0.67±0.20)cm.P<0.05)after intervention.Conclusions:The diameter of BCG scars was effectively correlated with PPD response,which indicates BCG scar formation may be an useful tool Io evaluate the effect of tuberculosis prevention.VA combined VD supplementation may play an immunoregulatory rale in BCG revuecination.This may contribute to the prevention of childhood tuberculosis.

Effects of Bacillus coagulans supplementation on the growth performance and gut health of broiler chickens with Clostridium perfringens-induced necrotic enteritis

Background: The poultry industry is in need of effective antibiotic alternatives to control outbreaks of necrotic enteritis(NE) due to Clostridium perfringens.Methods: This study was conducted to investigate the effects of feeding Bacil us coagulans on the growth performance and gut health of broiler chickens with C. perfringens-induced NE. Two hundred and forty 1-day-old broiler chicks were randomly assigned to a 2 × 2 factorial arrangement with two dietary B. coagulans levels(0 or 4 × 109 CFU/kg of diet) and two disease chal enge statuses(control or NE chal enged).Results: NE-induced reduction in body weight gain was relieved by the addition of B. coagulans into broiler diets compared with the NE-infected birds. NE infection damaged intestinal morphological structure, promoted intestinal C.perfringens growth and liver invasion, and enhanced anti-C. perfringens specific sI gA concentrations in the gut and specific IgG levels in serum compared with the uninfected birds. NE infection significantly(P < 0.05) decreased mucin-2(at 14 d post-infection(DPI), tol-like receptor 2(TLR2, at 7 and 14 DPI), TLR4(at 7 and 14 DPI), tumor necrosis factor super family15(TNFSF15, at 7 and 14 DPI), lysozyme(LYZ, at 14 DPI) and fowlicidin-2(at 7 and 14 DPI) mR NA levels, whereas it dramatical y(P = 0.001) increased IFN-γ mR NA levels at 7 DPI. However, chal enged birds fed diets supplemented with B.coagulans showed a significant(P < 0.01) decrease in gut lesion scores, decreased C. perfringens numbers in the cecum and liver, and an increase in fowlicidin-2 mR NA levels in compared with the uninfected birds. In addition, compared with the non-supplemented group, dietary inclusion of B. coagulans improved intestinal barrier structure, further increased specific sI gA levels and alkaline phosphatase(IAP) activity in the jejunum, enhanced the expression of jejunum lysozyme mR NA, and inhibited the growth, colonization, and invasion of C. perfringens; in contrast, it reduced serum-specific IgG concentrations and jejunum IFN-γ mR NA levels.Conclusion: These results indicated that dietary B. coagulans supplementation appeared to be effective in preventing the occurrence and reducing the severity of C. perfringens-induced NE in broiler chickens.