Cloning and sequence of the gene encoding a cefotaxime-hydrolyzing class A β-lactmase isolated from Escherichia coli

Basic Laboratory Procedures in Clinical Bacteriology

Background: A global problem of multi-drug resistance (MDR) among bacteria is the cause of hundreds of thousands of deaths every year. In response to the significant increase of MDR bacteria, legislative measures have widely been taken to limit or eliminate the use of antibiotics, including in the form of feed additives for livestock, but also in metaphylaxis and its treatment, which was the subject of EU Regulation in 2019/6. Numerous studies have documented that bacteria use both phenotypis and gentic strategies enabling a natural defence against antibiotics and the induction of mechanisms in increasing resistance to the used antibacterial chemicals. The mechanisms presented in this review developed by the bacteria have a significant impact on reducing the ability to combat bacterial infections in humans and animals. Moreover, the high prevalence of multi-resistant strains in the environment and the ease of transmission of drug-resistance genes between the different bacterial species including commensal flora and pathogenic like foodborne pathogens (E. coli, Campylobacter spp., Enterococcus spp., Salmonella spp., Listeria spp., Staphylococcus spp.) favor the rapid spread of multi-resistance among bacteria in humans and animals. Given the global threat posed by the widespread phenomenon of multi-drug resistance among bacteria which are dangerous for humans and animals, the subject of this study is the presentation of the mechanisms of resistance in most frequent bacteria called as “foodborne pathoges” isolated from human and animals. In order to present the significance of the global problem related to multi-drug resistance among selected pathogens, especially those danger to humans, the publication also presents statistical data on the percentage range of occurrence of drug resistance among selected bacteria in various regions of the world. In addition to the phenotypic characteristics of pathogen resistance, this review also presents detailed information on the detection of drug resistance genes for specific groups of antibiotics. It should be emphasized that the manuscript also presents the results of own research i.e., Campylobacter spp., E. coli or Enetrococcus spp. This subject and the presentation of data on the risks of drug resistance among bacteria will contribute to initiating research in implementing the prevention of drug resistance and the development of alternatives for antimicrobials methods of controlling bacteria.

/pdf/antibiotic-resistance-in-bacteria-a-review-3oluolqo.pdf

Antibiotic Resistance in Bacteria—A Review

https://doi.org/10.1016/j.psj.2022.101716

The control of poultry salmonellosis using organic agents: an updated overview

Abstract Background Microorganisms' survival is based on the importance of growth factors found in the microbial environment, as well as their overwhelming appetite for survival which is controlled by their genetic material. This study was designed to investigate the virulence patterns of Salmonella enterica serovar typhimurium associated with soft cheese. Two hundred and sixty (260) soft cheese samples were collected from vendors at Akure, Ikare and Owo in Ondo State, Nigeria. Molecular characterization of six (6) Salmonella enterica isolates for the possession of iro B, stn, hil A and spv C gene were revealed through multiplex polymerase chain reaction (PCR) analysis with DNA extraction, polymerase chain reaction (PCR) amplification, sequencing and genome blasting, respectively. Results Four (4) possess only iro B; two (2) possess only stn gene; and another three (3) possess hil A gene as none of the isolates possess spv C gene. This study revealed antibiotic-resistant Salmonella enterica serovar typhimurium is present in locally made soft cheese samples. It also revealed that virulence genes comprising hil A, stn and iro B as well as plasmids-mediating antibiotic resistance are present in S. enterica serovar typhimurium found in locally produced cheese. Conclusions The presence of molecularly elucidated virulence genes comprising Iro B , Stn and Hil A makes the soft cheese potentially viable for pathogenicity. This study recommends food-borne salmonellosis should be constantly monitored with appropriate cleaning of preparatory paraphernalia in cheese production. Constant sensitization of nomads to improve their awareness on milk-borne zoonosis and its associated risk factors is needed. 

/pdf/direct-detection-of-iro-b-stn-and-hil-a-virulence-genes-in-2f3v1onr.pdf

Direct detection of iro B, stn and hil A virulence genes in Salmonella enterica serovar typhimurium from non-ripened cheese

Aim: The study aimed to detect the invA gene in Salmonella isolated from milkfish in the Sidoarjo wet fish market. Materials and Methods: A total of 84 samples were prepared in enrichment media and isolated on the surface of Salmonella Shigella Agar. Salmonella growth produces transparent colonies with blackish color in the middle due to H2S gas formation. Samples were identified as Salmonella based on macroscopic colony morphology. Presumptive Salmonella sp. was put on Bismuth Sulfite Agar media. Salmonella was determined based on the results of the biochemical test that has been carried out using Microbact identification kits from negative gram staining.
Results: The results of this study indicate that 32 of 84 samples (38.09%) were Salmonella bacteria. Furthermore, the invA gene detection was carried out using the polymerase chain reaction technique. Electrophoresis results showed four positive samples contained invA gene with a length of 284 bp. Conclusion: Results in this study indicate that contamination of milkfish with Salmonella needs strict hygienic measures to prevent their transmission to human.

Keywords: human health, invA gene, milkfish, polymerase chain reaction, Salmonella

/pdf/detection-of-inva-gene-of-salmonella-from-milkfish-chanos-3kqkq9kg4t.pdf

Detection of invA gene of Salmonella from milkfish (Chanos chanos) at Sidoarjo wet fish market, Indonesia, using polymerase chain reaction technique

Aim: The research was to investigate the antibiotic resistance profile and to screen for methicillin-resistant Staphylococcus aureus (MRSA) from nasal mucosa swab of dogs

Materials and Methods: The samples were collected from three pet clinics, three K9 units, one veterinary teaching hospital, and one kennel in Surabaya Of the 50 total samples, 24 confirmed S aureus strains, which were used for antibiotic sensitivity tests using a disk diffusion method and screening of MRSA used oxacillin resistance screening for base (ORSAB)

Results: This study showed that there were differences in antibiotic resistance patterns among different locations Fourteen
isolates were screened for MRSA by culture on ORSAB

Conclusion: MRSA carriage was found on nasal swab of dogs, and dogs can act as reservoir of MRSA for spreading to human health

Keywords: antibiotic-resistant, dogs, methicillin resistant Staphylococcus aureus, Staphylococcus aureus

/pdf/profile-of-antibiotic-resistant-and-presence-of-methicillin-3icce83sgg.pdf

Profile of antibiotic-resistant and presence of methicillin-resistant Staphylococcus aureus from nasal swab of dogs from several animal clinics in Surabaya, Indonesia

Aim: This study was conducted to study the coagulase (coa) gene-based genetic diversity of Staphylococcus aureus, isolated from different samples of cattle from three different regions in East Java Province, Indonesia. Materials and Methods: A total of 160 raw milk samples collected in East Java Province, Indonesia, were screened for the presence of S. aureus. The presumptive isolates were confirmed by coa test. The confirmed S. aureus isolates were subjected to coa gene polymerase chain reaction. Results: Of 160 different samples, 20 (12.5%) isolates of S. aureus were confirmed by positive coa test. Of 20 S. aureus isolates, 19 (95%) isolates carried coa gene. Six different genotypes of coa gene, i.e., 440 bp, 510 bp, 547 bp, 680 bp, 740 bp, and 820 bp were obtained. One coa genotypes, 510 bp (10 isolates) were observed in polymorphism to be more prevalent than the others, and the genotype was present in at least one isolates from every region. Conclusion: It can be concluded that coa gene is easily epidemiological tool for detection of variation strain from S. aureus.

Keywords: coagulase gene, coagulase test, polymorphism, raw milk, Staphylococcus aureus

/pdf/detection-of-coagulase-gene-in-staphylococcus-aureus-from-2bh86pgs3m.pdf

Detection of coagulase gene in Staphylococcus aureus from several dairy farms in East Java, Indonesia, by polymerase chain reaction.

Klebsiella pneumoniae is one of 9 bacteria resistance to antibiotics in concern. This research aimed to detect any gene of blaTEM in bacteria of the K. pneumoniae isolated from swab of food-producing animals. In this study, 195 swab samples were taken from 17 sampling locations. Samples obtained were cultivated on selective medium and had several tests including identification, antibiotic sensitivity test using Kirby-Bauer method against antibiotics of ampicillin, cefotaxime, amoxicillin, meropenem, and trimetrophrim-sulfamethoxazole, and followed by PCR test for detecting the gene that was responsible for the antibiotic resistances. The results showed that 10 out of 195 samples were found to be K. pneumonia, those were 4 samples originated from dairy cows (SP-S1, SP-S3, SP-B2, SP-G4), 2 samples originated from beef (SPT-K1, SPT-K2), 1 sample originated from chickens (A-W5), and 3 samples originated from fish (IN-P2, IN-P3, IN-S3). Most of  isolates (9/10) were found to be resistant toward amoxicillin. These isolates were SP-S3, SP-B2, SP-G4, SPT-KI, SPT-K2, A-W5, IN-P2, IN-P3, and the IN-S3 and all of them also showed to be positive of blaTEM gene. It could be concluded that most of K. pneumoniae isolates from food animals harbour had Extended Spectrum Beta-Lactamase (ESBL) encoding gene.

/pdf/detection-of-blatem-gene-of-klebsiella-pneumoniae-isolated-1b2tam94rd.pdf

Detection of blaTEM Gene of Klebsiella pneumoniae Isolated from Swab of Food-Producing Animals in East Java

Ramandinianto SC, Khairullah AR, Effendi MH. 2020. MecA gene and methicillin resistant Staphylococcus aureus (MRSA) isolated from dairy farms in East Java, Indonesia. Biodiversitas 21: 3562-3568. Milk Borne Disease (MBD) can be caused by a variety of pathogenic bacteria, one of which is Staphylococcus aureus which has a large impact on aspects of public health. The therapy used to treat staphylococcal infection is Oxacillin preparations that can inhibit bacterial wall synthesis, but the adaptation of the mecA gene to staphylococcal cassette chromosome mec (SCCmec) causes the emergence of strains of methicillin-resistant S. aureus (MRSA). The purpose of this study was to detect the level of MRSA strain contamination in dairy cows in East Java by comparing the mecA gene, Oxacillin, and Cefoxitin Disc Diffusion Methods and Oxacillin Resistance Screen Agar (ORSA) detection methods. A total of 150 cow's milk samples were taken at 3 village dairy farms in East Java, samples were added to the enrichment media Buffer Pepton Water (BPW) and then isolates were planted and purified using Mannitol Salt Agar (MSA). The detection of MRSA was carried out by the Kirby Bauer disc diffusion preparation Cefoxitin 30 μg and Oxacillin 30 μg then confirmed by ORSA and the presence of mecA gene by the polymerase chain reaction (PCR) method. The results showed that from a total of 92 S. aureus isolates using Oxacillin disc test, 24 resistant isolates were obtained, using Cefoxitin disc test, 17 isolates were obtained, and using the ORSA test 18 MRSA isolates were obtained. MRSA isolates tested by PCR obtained evidence of 2 isolates of mecA gene. It can be concluded that the Oxacillin disc test was the highest sensitivity for detecting MRSA strain isolate, however, mecA gene was the golden standard to detect MRSA on the dairy farms

Keywords: Cefoxitin disc, milk-borne disease, mecA gene, MRSA, Oxacillin disc, ORSA

/pdf/meca-gene-and-methicillin-resistant-staphylococcus-aureus-bh485j8nht.pdf

Mustofa Helmi Effendi

Papers

Detection of invA gene of Salmonella from milkfish (Chanos chanos) at Sidoarjo wet fish market, Indonesia, using polymerase chain reaction technique

Profile of antibiotic-resistant and presence of methicillin-resistant Staphylococcus aureus from nasal swab of dogs from several animal clinics in Surabaya, Indonesia

Detection of coagulase gene in Staphylococcus aureus from several dairy farms in East Java, Indonesia, by polymerase chain reaction.

Detection of blaTEM Gene of Klebsiella pneumoniae Isolated from Swab of Food-Producing Animals in East Java

MecA gene and methicillin-resistant Staphylococcus aureus (MRSA) isolated from dairy farms in East Java, Indonesia