Biofilms and efflux pump regulatory gene (mexR) in multidrug-resistant Pseudomonas aeruginosa isolated from migratory birds in Egypt

Research (Published online: 19-10-2022)

8. Biofilms and efflux pump regulatory gene (mexR) in multidrug-resistant Pseudomonas aeruginosa isolated from migratory birds in Egypt

Esraa A. Elshafiee, Hanan S. Khalefa, Nayera M. Al-Atfeehy, Fatma Amer, Dalia A. Hamza, and Zeinab S. Ahmed

Veterinary World, 15(10): 2425-2431

ABSTRACT

Background and Aim: Multidrug-resistant (MDR) Pseudomonas aeruginosa is a global threat to public health. This study aimed to determine biofilms and efflux pump regulatory gene (mexR) in MDR P. aeruginosa isolates.

Materials and Methods: A total of 42 fecal samples of aquatic migratory birds collected during hunting season in Egypt were evaluated for the detection of P. aeruginosa according to standard culture-based methods. The antibiotic susceptibility of P. aeruginosa strains was evaluated using disk diffusion methods. The biofilm formation ability of the isolates was phenotypically determined using a colorimetric microtitration plate assay. Polymerase chain reaction amplification was performed to detect biofilm genes (PelA and PslA) and mexR.

Results: In total, 19 isolates (45.2%) were recovered from the 42 fecal samples of migratory birds. All isolates were identified as MDR P. aeruginosa, and 78.9% of the strains produced biofilms at different degrees. Molecular detection of biofilm extracellular polymeric substances revealed that PelA was the most predominant gene in the biofilm-producing isolates, followed by PslA. mexR was detected in 63.2% of MDR P. aeruginosa isolates, and its prevalence was higher in non–biofilm-producing strains (75%) than in biofilm-producing strains (60%).

Conclusion: Antibiotic resistance in P. aeruginosa isolates recovered from migratory birds through various mechanisms is a major public and animal health problem. It is important to consider the significance of migratory birds in disease transmission.

Keywords: biofilm, Egypt, mexR, migratory birds, multidrug-resistant, Pseudomonas aeruginosa.

Effects of parasites coinfection with other pathogens on animal host: A literature review

Review (Published online: 15-10-2022)

7. Effects of parasites coinfection with other pathogens on animal host: A literature review

Wael M. Hananeh, Asya Radhi, Rami M. Mukbel, and Zuhair Bani Ismail

Veterinary World, 15(10): 2414-2424

ABSTRACT

A parasite-host relationship is complicated and largely remained poorly understood, especially when mixed infections involving pathogenic bacteria and viruses are present in the same host. It has been found that most parasites are able to manipulate the host's immune responses to evade or overcome its defense systems. Several mechanisms have been postulated that may explain this phenomenon in different animal species. Recent evidence suggests that coinfections involving many parasitic species alter the host's vulnerability to other microorganisms, hinder diagnostic accuracy, and may negatively impact vaccination by altering the host's immune responsiveness. The objective of this review was to provide a comprehensive summary of the current understanding of how parasites interact with other pathogens in different animal species. A better understanding of this complex relationship will aid in the improvement efforts of disease diagnosis, treatment, and control measures such as novel and effective vaccines and therapeutics for infectious diseases.

Keywords: animals, coinfection, parasites.

Evolutionary conservation of motifs within vanA and vanB of vancomycin-resistant enterococci

Research (Published online: 13-10-2022)

6. Evolutionary conservation of motifs within vanA and vanB of vancomycin-resistant enterococci

Aylin Memili, Naseer Kutchy, Olubumi A. Braimah, and Olanrewaju B. Morenikeji

Veterinary World, 15(10): 2407-2413

ABSTRACT

Background and Aim: Global Health is threatened by the rapid emergence of multidrug-resistant bacteria. Antibiotic resistomes rapidly evolve, yet conserved motifs elucidated in our study have the potential for future drug targets for precision medicine. This study aimed to identify conserved genetic sequences and their evolutionary pathways among vancomycin-resistant Enterococcus species such as Enterococcus faecium and Enterococcus faecalis.

Materials and Methods: We retrieved a total of 26 complete amino acid and nucleotide sequences of resistance determinant genes against vancomycin (vanA and vanB), streptomycin (aac-aah), and penicillin (pbp5) from the publicly available genetic sequence database, GenBank. The sequences were comprised of bacteria classified under the genera of Enterococcus, Staphylococcus, Amycolatopsis, Ruminococcus, and Clostridium. Sequences were aligned with Clustal Omega Multiple Sequence Alignment program and Percent Identity Matrices were derived. Phylogenetic analyses to elucidate evolutionary relationships between sequences were conducted with the neighbor-end joining method through the Molecular Evolutionary Genetics Analysis (MEGAX) software, developed by the Institute of Molecular Evolutionary Genetics at Pennsylvania State University. Subsequent network analyses of the resistance gene, vanB, within E. faecium were derived from ScanProsite and InterPro.

Results: We observed the highest nucleotide sequence similarity of vanA regions within strains of E. faecium (100%) and E. faecalis (100%). Between Enterococcus genera, we continued to observe high sequence conservation for vanA and vanB, up to 99.9% similarity. Phylogenetic tree analyses suggest rapid acquisition of these determinants between strains within vanA and vanB, particularly between strains of Enterococcus genera, which may be indicative of horizontal gene transfer. Within E. faecium, Adenosine 5'-Triphosphate (ATP)-Grasp and D-ala-D-ala ligase (Ddl) were found as conserved domains of vanA and vanB. We additionally found that there is notable sequence conservation, up to 66.67%, between resistomes against vancomycin and streptomycin among E. faecium.

Conclusion: Resistance genes against vancomycin have highly conserved sequences between strains of Enterococcus bacteria. These conserved sequences within vanA and vanB encode for ATP-Grasp and Ddl motifs, which have functional properties for maintaining cell wall integrity. High sequence conservation is also observed among resistance genes against penicillin and streptomycin, which can inform future drug targets for broader spectrum therapies.

Keywords: antibiotic resistance, bioinformatics, Enterococcus, evolution, public health.

Risk factors of Bartonella spp. infection and the association between Bartonella spp. and T-lymphocyte subset alteration in asymptomatic retrovirus-infected cats in Bangkok Metropolitan, Thailand

Research (Published online: 13-10-2022)

5. Risk factors of Bartonella spp. infection and the association between Bartonella spp. and T-lymphocyte subset alteration in asymptomatic retrovirus-infected cats in Bangkok Metropolitan, Thailand

Krissda Boonaramrueng, Navapon Techakriengkrai, Channarong Rodkhum, and Rosama Pusoonthornthum

Veterinary World, 15(10): 2399-2406

ABSTRACT

Background and Aim: Cats are a reservoir for Bartonella spp. infection in humans. Human bartonellosis causes disseminated inflammation to develop in immunocompromised patients, such as those infected with human immunodeficiency virus. However, the associated risks of Bartonella spp. infection in immunocompromised retroviral-infected cats have been inconclusive. This study aimed to evaluate the associated risks of Bartonella spp. infection with the alteration of T-lymphocyte subsets of retroviral-infected cats.

Materials and Methods: We collected blood samples from 161 client-owned cats at veterinary clinics and hospitals throughout the Bangkok Metropolitan area from 2017 to 2020. The samples underwent hematological biochemical tests, feline retroviral status evaluation, Bartonella spp. polymerase chain reaction assay, immunofluorescence assay, and CD4+ and CD8+ lymphocyte counts. Risk factors associated with Bartonella spp. infection were determined by odds ratio (OR). Hematological and biochemical parameters were compared using independent t-tests. CD4+ and CD8+ lymphocyte counts and the CD4+/CD8+ ratio were compared among groups classified according to their retroviral and Bartonella spp. infection status.

Results: The prevalence of Bartonella spp. in our study cohort was 16.1%, and the seroprevalence was 94.9%. Cats aged >1 year were at a higher risk of seropositivity than cats aged <1 year (OR: 4.296, 95% confidence interval: 1.010–18.275). The CD8+ percentage was significantly higher in seropositive cats (p = 0.026). There was a significant reduction in the CD4+/CD8+ ratio between cats negative for both retrovirus and Bartonella spp. infection and cats with concurrent retrovirus and Bartonella spp. infection (p = 0.041).

Conclusion: In endemic countries or areas, cat owners must be made aware of the risk of exposure to Bartonella spp. due to the high rate of bacteremia and seroprevalence. Retrovirus-infected cats with concurrent Bartonella spp. infection also showed a significant, inverted CD4+/CD8+ ratio, which may be used as a novel marker in bartonellosis. Similar studies focusing on the different stages of retrovirus infection should be undertaken further to elucidate the effect of retrovirus infection on Bartonella spp. infection.

Keywords: Bartonella spp., cats, feline leukemia virus, feline immunodeficiency virus, retrovirus, risk factors, T-lymphocyte subsets.

Robusta coffee extracts inhibit quorum sensing activity in Chromobacterium violaceum and reduce biofilms against Bacillus cereus and Staphylococcus aureus

Research (Published online: 12-10-2022)

4. Robusta coffee extracts inhibit quorum sensing activity in Chromobacterium violaceum and reduce biofilms against Bacillus cereus and Staphylococcus aureus

Porwornwisit Tritripmongkol, Suthinee Sangkanu, Ratchadaporn Boripun, Juthatip Jeenkeawpieam, Julalak Chuprom, Veeranoot Nissapatorn, Maria de Lourdes Pereira, Alok K. Paul, and Watcharapong Mitsuwan

Veterinary World, 15(10): 2391-2398

ABSTRACT

Background and Aim: Bacillus cereus and Staphylococcus aureus cause foodborne intoxication in humans and animals. Pathogens can produce biofilms controlled by the quorum sensing system. The study aimed to investigate the antibacterial, antibiofilm, and anti-quorum sensing activities of Coffea canephora P. ex Fr. (Robusta coffee) extracts against B. cereus and S. aureus.

Materials and Methods: Ethanol extracts of fruit peels and seeds of Robusta coffee were tested for antibacterial activity against B. cereus and S. aureus using a broth microdilution assay. Reduction of the biofilm formation and elimination of the viability of mature biofilm-grown cells of B. cereus and S. aureus were determined. Inhibition of quorum sensing activity in Chromobacterium violaceum by the extracts was investigated using the disk diffusion method and flask incubation assay.

Results: Fresh fruit peel extract showed the strongest antibacterial activity against B. cereus and S. aureus with minimum inhibitory concentration (MIC) values of 2 and 4 mg/mL, respectively. However, the extracts did not inhibit Escherichia coli, avian pathogenic E. coli, and Pseudomonas aeruginosa at 8 mg/mL. Significant inhibition of biofilm formation at 1/2 × MIC of the fresh peel extract was detected in B. cereus (56.37%) and S. aureus (39.69 %), respectively. At 8 × MIC of the fresh peel extract, a significant elimination of the mature biofilm viability was detected in B. cereus (92.48%) and S. aureus (74.49%), respectively. The results showed that fresh and dried peel fruit extracts at 1/2 × MIC significantly reduced violacein production with the highest percentage inhibition ranging from 44.53 to 47.48% at 24 h (p ≤ 0.05).

Conclusion: The results of the present study suggest the potential therapeutic benefits of Robusta coffee extracts in inhibiting the growth, biofilm, and quorum sensing of both B. cereus and S. aureus. The results put forward an alternative strategy to control the foodborne intoxications caused by both pathogens.

Keywords: Bacillus cereus, biofilms, quorum sensing, Robusta coffee extract, Staphylococcus aureus.

Effect of thioredoxin on the immunogenicity of the recombinant P32 protein of lumpy skin disease virus

Research (Published online: 11-10-2022)

3. Effect of thioredoxin on the immunogenicity of the recombinant P32 protein of lumpy skin disease virus

Kanat Tursunov, Laura Tokhtarova, Darkhan Kanayev, Raikhan Mustafina, and Kanatbek Mukantayev

Veterinary World, 15(10): 2384-2390

ABSTRACT

Background and Aim: The rapid spread of lumpy skin disease (LSD) globally poses a serious threat to the agricultural sector. The timely and accurate diagnosis of the disease is crucial to control LSD. This study aimed to determine the effect of thioredoxin on the immunogenicity of the recombinant P32 (rP32) protein of LSD virus (LSDV). Since the P32 protein is poorly soluble, it is often expressed by adding an auxiliary sequence of a highly soluble partner protein such as thioredoxin.

Materials and Methods: The P32 gene fragment was amplified using a polymerase chain reaction from genomic DNA used as a template. The resulting DNA fragments were cloned into the pET32a vector, and transformed into Escherichia coli BL21 (DE3) cells through electroporation. Purification of the rP32 protein was performed using a HisTrap column. Purified rP32 protein fused with thioredoxin (rP32Trx) was characterized by western blotting, liquid chromatography with tandem mass spectrometry and indirect enzyme-linked immunosorbent assay (ELISA).

Results: Indirect ELISA revealed that, despite the lower molecular weight, the main part of the antibodies in the serum of immunized mice was directed against thioredoxin and not the target P32 protein. Thus, the antibody titers against rP32Trx were 1:102400, whereas antibody titers against heterologous recombinant 3BTrx and PD1Trx proteins were 1:25600 and 1:51200, respectively. Concurrently, the antibodies did not bind to the heterologous recombinant PD1 protein, which did not contain thioredoxin.

Conclusion: The results showed that the rP32 protein fused with the partner protein thioredoxin could not be used to obtain polyclonal and monoclonal antibodies. However, the recombinant fusion protein rP32Trx can be used to develop a serological test to detect antibodies, since antibodies against thioredoxin were not detected in the animal sera.

Keywords: immunogenicity, lumpy skin disease virus, P32 protein, recombinant antigen, thioredoxin.

Expression and epitope prediction of MPT64 recombinant proteins from clinical isolates of Mycobacterium tuberculosis as immunoserodiagnostic candidates

Research (Published online: 08-10-2022)

2. Expression and epitope prediction of MPT64 recombinant proteins from clinical isolates of Mycobacterium tuberculosis as immunoserodiagnostic candidates

Fihiruddin Fihiruddin, Nurul Inayati, Raudatul Jannah, Lalu Unsunnidhal, and Asmarani Kusumawati

Veterinary World, 15(10): 2376-2383

ABSTRACT

Background and Aim: The success in the handling and prevention of tuberculosis (TB) cases is highly dependent on their rapid detection, monitoring, and treatment. The efficacy of the Bacille Calmette–Guerin (BCG) vaccine is inconclusive in eastern Indonesia. The RV1980c gene of Mycobacterium tuberculosis encodes an antigenic protein that is considered to be a virulence factor, as it can stimulate the immune response in patients with TB. This study aimed to study the expression and epitope indicator of MPT64 recombinant proteins from clinical isolates of M. tuberculosis as immunoserodiagnostic candidates for pET SUMO plasmids from clinical isolates as candidates for serodiagnostic tests and recombinant vaccines.

Materials and Methods: The polymerase chain reaction (PCR) product of the RV1980c gene was inserted into the SUMO pET plasmid, which was then transformed into Escherichia coli BL21 (DE3) cells and expressed in Luria Bertani media induced by 1.0 M IPTG. Subsequently, sequencing was performed and the results were analyzed using the ClustalW and National Center for Biotechnology Information BLAST software. The T-cell epitope prognosis was then explained by GENETYX version 8.0., for the prediction of B-cell epitope, as assessed using an Immune Epitope Database analysis.

Results: The PCR product of the RV1980c gene had a length of 619 bp. Moreover, SDS–polyacrylamide gel electrophoresis and Western blotting revealed that the protein encoded by the Rv1980c gene weighed 36 kDa. We gained nine specific T-cell epitopes according to Iad Pattern position and eight epitopes according to Rothbard/Taylor Pattern Position; furthermore, we detected five B-cell epitopes in the RV1980c gene.

Conclusion: The MPT64 protein encoded by the RV1980c gene carries epitopes that are realized by lymphocytes and represent potential immunoserodiagnostic candidates in diagnostic immunology.

Keywords: Escherichia coli, immune response, lymphocytes, tuberculosis.