Genomic diversity and nutritional analysis of multi-drug resistant extended spectrum β-lactamase Producing-Klebsiella pneumoniae genes isolated from mastitic cattle milk in district peshawar, Pakistan
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SUBMITTER: Saddam
PROVIDER: S-EPMC11337036 | biostudies-literature | 2024 Aug
REPOSITORIES: biostudies-literature
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Project description:Klebsiella pneumoniae is a major zoonotic pathogen that causes a variety of severe illnesses as well as mastitis. The distribution of mastitis-causing K. Pneumoniae and its virulence factors vary by country and geographical location. The present study aimed to find out the occurrence of Multidrug-resistant (MDR) K. Pneumoniae and their capsular resistance genes which were undocumented previously in cow farms of district Peshawar, Pakistan. A total of 700 milk samples from symptomatic mastitic cows were screened for MDR K. Pneumoniae. Furthermore, the characterization of capsular resistance genes was done by molecular techniques. Among these samples, K. pneumoniae was found 180/700 (25.7%), while MDR K. pneumoniae was found 80/180 (44.4%). The antibiogram analysis revealed high resistance to Vancomycin (95%) while highly sensitive to Ceftazidime (80%). The distribution of capsular genes shows the most common serotype K2 gene 39/80 (48.7%), followed by serotype K1 gene 34/80 (42.5%), serotype K5 17/80 (21.2%), and serotype K54 13/80 (16.2), respectively. Moreover, the co-occurrence of serotypes K1+K2 was found at 11.25%, KI+K5 was 05%, K1+K54 was 3.75%, and K2+K5 was 7.5%, respectively. A statistically significant association (p ≤ 0.05) was found between predicted and discovered K. pneumoniae values. In conclusion, the presence of MDR K. pneumoniae in combination with capsular genes may be a possible threat to dairy farm animals and humans in Peshawar, Pakistan. It may give us special attention to follow up on hygienic practices in livestock management.
| S-EPMC9970052 | biostudies-literature
Project description:BackgroundSahiwal cattle is an indigenous cattle breed of Pakistan and mastitis is one of the major problems faced by Sahiwal cattle which hinders its production potential. The study was designed to investigate the milk microbiota of healthy and mastitic Sahiwal cattle as part of a multistep project to develop probiotics for the mitigation and control of mastitis. Milk samples of Sahiwal cattle (healthy clinical mastitis and subclinical mastitis) reared under similar husbandry and management practices were processed for 16S rRNA gene base metagenomics analysis.ResultsResults revealed that Proteobacteria were dominant in the healthy group and subclinical mastitis group (56.48% and 48.77%, respectively) as compared to the clinical mastitis group (2.68%). In contrast, Firmicutes were abundant in the clinical mastitis group (64%) as compared to the healthy and subclinical mastitis groups (15.87% and 38.98%, respectively). Dominant species assigned in the healthy group were Ignavibacterium album, Novosphingobium capsulatum, Akkermansia muciniphila and Lactobacillus fermentum.The clinical mastitis group was dominated by Streptococcus dysgalactiae and Corynebacterium bovis, while subclinical mastitis group included Lactobacillus fermentum and uncultured acidobacteriales and Akkermansia muciniphila as dominant species. Alpha diversity indices showed higher microbial diversity in the healthy group compared to the clinical and sub-clinical mastitis groups.ConclusionIt is concluded that the milk microbiota of healthy sahiwal cattle has higher diversity and dominant taxa in the different groups may be used as signature microbes for mastitis susceptibility. Akkermansia muciniphila is one of candidate specie that was identified and may be used for development of probiotics.
| S-EPMC10594912 | biostudies-literature
Project description:BackgroundBovine mastitis is a common udder disease in cattle, mainly caused by bacteria and other infectious agents. Traditionally antibiotics are used for their treatment, but the development of antibiotic resistance has increased the importance of using non antibiotic alternative such as probiotic. In current study a previously in vitro characterized isolate Lactobacillus plantarum CM49 infused into two groups of cattle suffering from clinical mastitis (n = 5) and sub-clinical mastitis (n = 5).ResultsThe bacterial composition and diversity analysis of milk samples before and after probiotic administration was analyzed using 16S rRNA gene base metagenomic analysis and lactobacillus counts were also evaluated using Real time PCR. The results show that there was an increase in abundance of Proteobacteria and decrease in Firmicutes at phylum level in both groups while major mastitogens genera Staphylococcus and Streptococcus abundance was reduced after treatment in sub-clinical mastitis group (SCMG) and clinical mastitis group (CMG) respectively. Lactobacilli counts evaluated through Real time PCR showed an increase in number, furthermore diversity indices showed an increase in diversity after treatment with probiotic.ConclusionIt is concluded from the results that Lactobacillus plantarum CM49 may serve as promising candidate for improving dysbiosis resulting from mastitis and improving microbial diversity.
| S-EPMC11871703 | biostudies-literature
Project description:The emergence and spread of antimicrobial resistance genes and resistant bacteria do not recognize animal, human, or geographic boundaries. Addressing this threat requires a multidisciplinary approach involving human, animal, and environmental health (One Health) sectors. This is because antimicrobial agents used in veterinary medicine have been reported to be the same or similar to those in human medicine use. Extended-spectrum β-lactamase (ESBL) E. coli is a growing public health problem worldwide, and the agri-food industry is increasingly becoming a source of clinically important ESBL bacteria. Accordingly, the aim of this study was to investigate the occurrence and characteristics of ESBL-producing E. coli from dairy cattle, milk, and the farm environment. E. coli isolates were identified by their 16sRNA, and their ESBL production was confirmed using ESBL-CHROMagar media and the double disk diffusion method. Genotypes of ESBL producers were characterized using multiplex polymerase chain reaction (mPCR) assay. It was found that 18 (4.8%) of the total samples were positive for ESBL-producing E. coli. Of these, 66.7% were from milk, and 27.8% and 5.5% were from the farm environment and faecal samples, respectively. Predominant ESBL genotypes identified were a combination of both TEM and CTX-M in eight out of 18 (44.4%) isolates. Four (22.2%) isolates produced the CTX-M gene, two (11.1%) isolates produced the TEM gene, and four (22.2%) remaining isolates produced the ESBL genes other than TEM, SHV, CTX-M, and OXA. The SHV and OXA gene were not detected in all 18 isolates. In all, there were four profiles of genetic similarity. The occurrence of these genotypes in indicator organisms from dairy cattle, milk, and the farm environment further re-enforced the potential of food-animals as sources of ESBL-producing E. coli infection in humans via the food chain. Thus, there is the need for the adoption of a tripartite One Health approach in surveillance and monitoring to control antimicrobial resistance.
| S-EPMC7760176 | biostudies-literature
Project description:IntroductionThe rising prevalence of Extended-Spectrum Beta-Lactamase (ESBL)-producing Klebsiella species (spp.) poses a significant threat to human and animal health and environmental safety. To address this pressing issue, a comprehensive study was undertaken to elucidate the burden and dissemination mechanisms of ESBL-Klebsiella spp. in dairy cattle farms.MethodsFifty-seven Klebsiella species were isolated on CHROMagar™ ESBL plates and confirmed with MADLI-TOF MS and whole genome sequenced from 14 dairy farms.Results and discussionSix families of beta-lactamase (bla) (bla CTX-M, bla SHV, bla TEM, bla OXY, bla OXA, and bla SED) were detected in ESBL-Klebsiella spp. genomes. Most (73%) of isolates had the first three types of beta-lactamase genes, with bla SHV being the most frequent, followed by bla CTX-M. Most (93%) isolates harbored two or more bla genes. The isolates were genotypically MDR, with 26 distinct types of antibiotic resistance genes (ARGs) and point mutations in gyrA, gyrB, and parC genes. The genomes also harbored 22 different plasmid replicon types, including three novel IncFII. The IncFII and Col440I plasmids were the most frequent and were associated with bla CTXM-27 and qnrB19 genes, respectively. Eighteen distinct sequence types (STs), including eight isolates with novel STs of K. pneumoniae, were detected. The most frequently occurring STs were ST353 (n = 8), ST469 (n = 6), and the novel ST7501 (n = 6). Clusters of ESBL-Klebsiella strains with identical STs, plasmids, and ARGs were detected in multiple farms, suggesting possible clonal expansion. The same ESBL variant was linked to identical plasmids in different Klebsiella STs in some farms, suggesting horizontal spread of the resistance gene. The high burden and dual spread mechanism of ESBL genes in Klebsiella species, combined with the emergence of novel sequence types, could swiftly increase the prevalence of ESBL-Klebsiella spp., posing significant risks to human, animal, and environmental health. Immediate action is needed to implement rigorous surveillance and control measures to mitigate this risk.
| S-EPMC11458399 | biostudies-literature
Project description:Background. Klebsiella pneumoniae is part of the ESKAPE (Enterococcus faecium, Staphylococcus aureus, K. pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.) group of multidrug-resistant (MDR) pathogens. K. pneumoniae is the leading cause of antimicrobial resistance-associated mortality and the second leading cause of nosocomial bloodstream infections (BSIs), globally and in sub-Saharan Africa. Therefore, it was aimed to determine the antibiotic resistance patterns of K. pneumoniae isolated from blood cultures of patients with features of sepsis at Mulago National Referral Hospital, Uganda. Methods. The cross-sectional study on patients with features of sepsis utilized K. pneumoniae (n=30) isolated from positive blood culture specimens. The antibiotic resistance profile was determined by the Clinical and Laboratory Standards Institute's Kirby-Bauer disc diffusion method, which was used to classify the isolates as susceptible, intermediate and resistant. K. pneumoniae isolates that were resistant to third-generation cephalosporins were subjected to extended-spectrum β-lactamase (ESBL) screening and confirmation using the double-disc synergy test using cefotaxime, ceftazidime, ceftriaxone, cefotaxime-clavulanic acid and ceftazidime-clavulanic acid. The results were analysed for frequencies. Results. K. pneumoniae isolates showed emerging resistance to imipenem at 13% (4 out of 30) followed by amikacin at 17% (5 out of 30). There was intermediate resistance to gentamycin at 60% (18 out of 30). However, K. pneumoniae showed the highest resistance to piperacillin at 100% (30 out of 30) followed by sulphamethoxazole-trimethoprim and cefepime, both showing a percentage of 97% (29 out of 30). Up to 16 out of 30 (53.3%) of K. pneumoniae were positive for ESBL production, whilst 14 out of 30 (46.7%) were negative. Conclusion. There was a high prevalence of antibiotic-resistant ESBL-producing K. pneumoniae isolates from BSI of patients with features of sepsis in Uganda's Mulago National Referral Hospital.
| S-EPMC11840160 | biostudies-literature
Project description: Not available
| S-EPMC10658008 | biostudies-literature
Project description:BackgroundExtended-spectrum β-lactamase (ESBL) production is the major resistance mechanism to β-lactam antibiotics in Enterobacteriaceae. In addition, emergence of plasmid-mediated quinolone resistance (PMQR) in ESBL-producing isolates has become a global threat for treatment of these infections.ObjectivesWe investigated the association between ESBL production and quinolone resistance in urinary isolates of K. pneumoniae.Patients and methodsA total of 196 urinary isolates of K. pneumoniae were collected from Imam Hussein Hospital in Tehran during a four year period (2008-2012). Antibiotic susceptibility was determined by disc diffusion and ESBL production was screened using the phenotypic confirmatory test (PCT).ResultsAll isolates were susceptible to imipenem. Resistance to piperacillin and cefotaxime were 66.3% and 50.5%, respectively. Resistance to ceftazidime, amoxiclave, aztreonam, ceftriaxone, cefepime, nitrofurantoin, gentamicin, ciprofloxacin, nalidixic acid, ofloxacin, norfloxacin, levofloxacin, amikacin and pipracilin/tazobactam were less than 50%. ESBL production was detected in 92 isolates (46.9%) of which, 61.9% were resistant to nalidixic acid and 65.2% to ciprofloxacin. Multidrug-resistance was observed in 96.7% of ESBL producers.ConclusionsOur results showed coexistence of ESBL and quinolone resistance in the majority of the uropathogenic K. pneumoniae test isolates suggesting that care should be taken for the choice of antibiotic therapy.
| S-EPMC4217673 | biostudies-literature
Project description:BackgroundKlebsiella species are ubiquitous in nature and can be found in the natural environment and on mucosal surfaces of mammals and it is an important multidrug-resistant pathogen affecting humans and is a major source for hospital acquired infections. The aim of this study is to investigate the prevalence of ESBL enzyme among Klebsiella pneumoniae isolates by phenotypic methods from different hospital wards and detection of ESBL resistance genes such as TEM and SHV in Sulaimani city/ Kurdistan-Iraq.MethodsKlebsiella pneumoniae isolates were collected from different clinical samples from different hospitals, the isolates were identified by standard technique. Screening of ESBLs was undertaken by using double disk diffusion and standard disk diffusion methods. Real-time PCR was used for genotypic detection of TEM and SHV genes according to the standard protocol.ResultOut of 54 Klebsiella pneumoniae isolates; 28 were ESBL positive, The pattern of antimicrobial susceptibility testing showed that the most resistant antibiotic are AMP (100%), AMC (100%) followed by CAZ (83.33%), CTX (75.9%), CPM (74%), ATM (70.37%). Both TEM and SHV genes were detected among 28 (51.85%) ESBL positive by using Real-time PCR method.ConclusionSHV gene was detected in most of the isolates of ESBL producers of Klebsiella pneumoniae.
| S-EPMC9522270 | biostudies-literature
Project description:A variety of proteases release hundreds of endogenous peptide fragments from intact bovine milk proteins. Mass spectrometry-based peptidomics allows for high throughput sequence assignment of a large number of these peptides. Mastitis is known to result in increased protease activity in the mammary gland. Therefore, we hypothesized that subclinically mastitic milks would contain higher concentrations of released peptides. In this work, milks were sampled from three cows and, for each, one healthy and one subclinically mastitic teat were sampled for milk. Peptides were analyzed by nano-liquid chromatography quadrupole time of flight tandem mass spectrometry and identified with database searching. In total, 682 peptides were identified. The total number of released peptides increased 146% from healthy to subclinically mastitic milks (p < 0.05), and the total abundance of released peptides also increased significantly (p < 0.05). Bioinformatic analysis of enzyme cleavage revealed increases in activity of cathepsin D and elastase (p < 0.05) with subclinical mastitis.
| S-EPMC4402940 | biostudies-literature