Volume 15 | October
Research Article | 07 Oct 2022
Comparative immune responses after vaccination with the formulated inactivated African horse sickness vaccine serotype 1 between naïve horses and pretreated horses with the live-attenuated African horse sickness vaccine
Narongsak Chaiyabutr, Suphot Wattanaphansak, Rachod Tantilerdcharoen, Surasak Akesowan, Suraseha Ouisuwan, and Darm Naraporn

Volume-15 | Issue-10 | Article-1 | https://doi.org/10.14202/vetworld.2022.2365-2375

Preview Abstract
African horse sickness (AHS) is a non-contagious, high mortality, and insect-borne disease caused by a double-stranded RNA virus from the genus Orbivirus. The study aimed to develop inactivated vaccines serotype 1 inactivated AHS vaccine (IAV) and to compare the effect of IAV on antibody responses in young naïve horses and adult horses pre-immunized with live-attenuated AHS virus (AHSV) serotypes 1, 3, and 4 live-attenuated vaccine (LAV). A total of 27 horses were vaccinated in two trials. Twelve AHS naïve young horses and 15 adult horses were divided into three groups of 4 and 5 horses each, respectively. Horses in control Group 1 were treated with phosphate-buffered saline. Horses in Group 2 were subcutaneously vaccinated with 2 mL of formulated IAV with 10% Gel 01™ (Seppic, France) on day 0 and horses in Group 3 were subcutaneously vaccinated with 2 mL of IAV on day 0 and a booster on day 28. The IAV vaccine was prepared by isolating the AHSV serotype 1 growing on Vero cells, 10× virus titer was concentrated by ultrafiltration and chemically killed by formalin, using 10% Gel 01™ as an adjuvant. Ethylenediaminetetraacetic acid blood samples were taken for hematology, blood biochemistry, and antibody titers using an immunoperoxidase monolayer assay on 158th day post-vaccination. Vaccination with IAV serotype 1 in adult horses pretreated with LAV increased antibody titers more than in young naïve vaccinated horses. The total leukocyte count and %neutrophils significantly increased, while %lymphocytes and %eosinophils significantly decreased on day 1 after vaccination; no local reactions were observed at the site of injection in any group. All biochemical and electrolyte analyte values were within the normal range after vaccination. The formulation of IAV serotype 1 using Gel 01™ as an adjuvant is safe and induces high antibody titers. This IAV formulation induced a high antibody response in horses without causing local reactions and mild systemic effects. However, AHS naïve horses still required ≥2 vaccinations and an annual booster vaccination to achieve high antibody titers. Keywords: African horse sickness, inactivated vaccine, naïve young horse.
Research Article | 08 Oct 2022
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

Volume-15 | Issue-10 | Article-2 | https://doi.org/10.14202/vetworld.2022.2376-2383

Preview Abstract
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. 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. 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. 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.
Research Article | 11 Oct 2022
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

Volume-15 | Issue-10 | Article-3 | https://doi.org/10.14202/vetworld.2022.2384-2390

Preview Abstract
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. 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). 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. 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.
Research Article | 12 Oct 2022
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

Volume-15 | Issue-10 | Article-4 | https://doi.org/10.14202/vetworld.2022.2391-2398

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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. 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. 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). 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.

Volume-15 | Issue-10 | Article-5 | https://doi.org/10.14202/vetworld.2022.2399-2406

Preview Abstract
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. 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. 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). 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.
Research Article | 13 Oct 2022
Evolutionary conservation of motifs within vanA and vanB of vancomycin-resistant enterococci
Aylin Memili, Naseer Kutchy, Olubumi A. Braimah, and Olanrewaju B. Morenikeji

Volume-15 | Issue-10 | Article-6 | https://doi.org/10.14202/vetworld.2022.2407-2413

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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. 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. 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. 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.
Review Article | 15 Oct 2022
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

Volume-15 | Issue-10 | Article-7 | https://doi.org/10.14202/vetworld.2022.2414-2424

Preview 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.
Research Article | 19 Oct 2022
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

Volume-15 | Issue-10 | Article-8 | https://doi.org/10.14202/vetworld.2022.2425-2431

Preview Abstract
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. 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. 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%). 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.
Research Article | 20 Oct 2022
First study on the immunohistochemical expression of cyclooxygenase-2 and clinicopathological association in canine hepatoid gland neoplasms
Pinkarn Chantawong, Thanongsak Mamom, Sahatchai Tangtrongsup, Setthakit Chitsanoor, and Hassadin Boonsriroj

Volume-15 | Issue-10 | Article-9 | https://doi.org/10.14202/vetworld.2022.2432-2441

Preview Abstract
Hepatoid gland neoplasms (HGNs) constitute one of the most common cutaneous tumors that arise from perianal glands in dogs and are clinically characterized by rapid growth. Cyclooxygenase-2 (COX-2), the inducible form of the enzyme, is associated with several hallmarks of tumorigenesis. Its expression has been confirmed in several human and animal neoplastic tissues, but there are no reports in hepatoid gland tissues. Therefore, this study aimed to investigate COX-2 immunoexpression in canine HGNs, compare the expression among groups of normal hepatoid glands, hepatoid gland adenomas (HGAs), hepatoid gland epitheliomas (HGEs), and hepatoid gland carcinomas (HGCs), and assess the association of the COX-2 expression with clinicopathological features. Sixty-one formalin-fixed paraffin-embedded canine hepatoid gland tissues (20 samples of HGAs, 16 of HGEs, 15 of HGCs, and 10 of normal hepatoid glands) were analyzed for COX-2 expression using immunohistochemistry with scoring for percentage positivity and intensity. Multiple comparisons of COX-2 expression among normal and neoplastic hepatoid glands and the associations between COX-2 expression and clinicopathological features were analyzed. Cyclooxygenase-2 expression was not detected in 60% of normal hepatoid glands and 25% of HGAs. Seventy-five percent of HGAs had a weak expression, while 43.7% and 56.3% of HGEs showed weak and moderate expression, respectively. The expression of HGCs ranged from weak (13.3%) to moderate (33.3%) and strong (53.3%). The immunoreactivity score of COX-2 labeling was significantly different among the normal and neoplastic hepatoid glands (p < 0.0001). The highest score was observed in the HGCs. Only in HGCs, the strong COX-2 expression was significantly associated with some clinicopathological features, including tissue invasion (p = 0.007) and necrosis (p = 0.029). These results suggest that COX-2 may play a role in the modulation of neoplastic cell growth. These preliminary data lead to further investigation on the potential of COX-2 expression as a prognostic indicator and COX-2 inhibitors for canine HGCs treatment. Keywords: canine, clinicopathological features, cyclooxygenase-2, hepatoid gland neoplasias, immunohistochemistry.
Review Article | 21 Oct 2022
Prevalence and resistance to gastrointestinal parasites in goats: A review
Takalani Judas Mpofu, Khathutshelo Agree Nephawe, and Bohani Mtileni

Volume-15 | Issue-10 | Article-10 | https://doi.org/10.14202/vetworld.2022.2442-2452

Preview Abstract
Gastrointestinal parasitism, particularly nematode infection, is a major health issue affecting goats worldwide, resulting in clinical diseases and productivity loss. Prevalent gastrointestinal parasites (GIPs) affecting goats in South Africa are the Strongyloides papillosus, Eimeria spp., and Strongyles, especially the Haemonchus contortus and Trichostrongylus spp. According to the issues discussed in this paper and by other authors, the prevalence and intensity of various GIPs vary with an animal's location, breed, age, sex, and season. Because GIPs easily develop resistance to chemical treatment, selecting and breeding genetically GIP-resistant animals would be a relatively simple and inexpensive strategy for reducing or eliminating the current reliance on chemotherapy. Potential phenotypic indicators for selecting GIP-resistant goats include parasitological, immunological, and pathological phenotypic markers. Synergistic use of these indicators should be encouraged for a more accurate simplified genotype selection of resistant animals. Genes with Mendelian inheritance, particularly those involved in immunoregulatory mechanisms, have been identified in goats. Exploring this knowledge base to develop cost-effective molecular tools that facilitate enhanced genetic improvement programs is a current challenge. Future statistical and biological models should investigate genetic variations within genomic regions and different candidate genes involved in immunoregulatory mechanisms, as well as the identification of single nucleotide polymorphisms known to affect GIP infection levels.
Research Article | 22 Oct 2022
The effect of feeding black soldier fly larvae on growth performance, protein, and fat content of red hybrid tilapia (Oreochromis spp.)
H. N. Aisyah, Z. A. R. Athirah, W. R. Hanani, S. S. Arshad, H. A. Hassim, M. F. Nazarudin, and M. Y. Ina-Salwany

Volume-15 | Issue-10 | Article-11 | https://doi.org/10.14202/vetworld.2022.2453-2457

Preview Abstract
In the aquaculture industry, the crucial goal is to minimize production costs, especially feeding costs, without significant side effects. Black soldier fly larva (BSFL) is a locally available, eco-friendly, and sustainable source that is high in crude protein (42% dry matter [DM]) and fat (35% DM). This study aimed to determine the growth performance along with the composition of crude fat and protein in red hybrid fingerlings after the addition of BSFL into the diet. A total of 120 fingerlings of uniform size (mean initial weight of 1.46 ± 0.06 g) were randomly assigned to one of four groups (n = 10) (A, B, C, and D) per tank (1 m × 2 m × 1 m). For 21 days, Group A (control group) was fed with 100% commercial diet; Group B was fed with 90% commercial fish diet + 10% BSFL; Group C was fed with 80% commercial fish diet + 20% BSFL; and Group D was fed with 70% commercial fish diet + 30% BSFL. Feed efficiency, growth performance, and proximate composition analysis were performed on the fish. The results displayed that the group with the highest BSFL percentage had a greater effect on protein and fat composition than the control group. The proximate composition analysis of fish-fed diet revealed that an increase in the level of BSFL inclusion increases the protein content in the fish. In comparison to the other groups, the experimental diet with 30% BSFL inclusion has the highest levels of crude protein (80.30% DM) and fat (2.90% DM). It is concluded that incorporating BSFL into a commercial diet for red hybrid tilapia fingerlings increased crude protein and fat composition, providing an alternative protein and fat source in fish diets. Keywords: black soldier fly larvae, growth performance, Oreochromis spp., red hybrid tilapia.
Research Article | 22 Oct 2022
Interaction of Cyprinus carpio Linnaeus with the biofilm-forming Aeromonas hydrophila
Ekaterina Lenchenko, Svyatoslav Lenchenko, Nadezhda Sachivkina, Olga Kuznetsova, and Alfia Ibragimova

Volume-15 | Issue-10 | Article-12 | https://doi.org/10.14202/vetworld.2022.2458-2465

Preview Abstract
The resistance of susceptible fish populations and the adaptive potential of heterogeneous biofilms, which cause multiple antibacterial resistance and long-term persistence of microorganisms, mediate the development and outcome of the infectious process. The study of the fish immunological parameters in interaction with biofilm-forming bacteria is of practical importance for assessing the stability of the homeostasis of the fish. This study aimed to determine the immunobiological parameters of Cyprinus carpio Linnaeus when interacting with biofilm-forming bacteria Aeromonas hydrophila. Clinically healthy fish C. carpio L. (Linnaeus, 1758) of both sexes, aged 4 years, and weighing 1.0–1.5 kg (n = 10), were used in this study. The fish were taken from the pond of the VNIIR experimental base in the period of 2020–2022. The standard method was employed to determine the phagocytic activity of blood cells, the total redox activity of neutrophils, and the bactericidal activity of blood serum. After 24–48 h of cultivation in nutrient broth, the implementation of the processes of intercellular communication of bacteria had common patterns of formation of the heterogeneous structure of biofilms. Moreover, analyzing the optical density indices (density, D), it was observed that A. hydrophila was a strong producer of biofilms, as the optical density of the sample (density of sample, Ds) exceeded the optical density of the control (density of control, Dc) by more than 4 times (D = 0.464 ± 0.07). The ratio of the average number of microorganisms attached to the surface of one erythrocyte (average adhesion index) and the percentage (%) of erythrocytes having bacteria on their surface (adhesion coefficient [AC]) was 14.05 ± 0.72, and the adhesion index, AI was ≥4.00, indicating A. hydrophila to be highly adhesive. In addition, the AC of erythrocytes having bacteria on the surface was 14.05% ± 0.72%. A direct correlation was established (R2 = 0.94) between the AC (14.05% ± 0.11%–13.29% ± 0.08%) and the phagocytic index (11.3% ± 0.29%–32.0% ± 0.8%). The indicators of spontaneous nitro blue tetrazolium were 103.20 ± 11.70 when estimating the total redox activity of neutrophils. The optical density increased to 182.10 ± 21.12 with the addition of 20.0 μL of A. hydrophila bacteria (1 billion/mL) and the activity of neutrophils also increased. Among the markers of homeostasis stability, immunological indicators most fully reflect the mechanisms of initiation, development, and outcome of the infectious process mediated by the interaction of adhesive molecules of multicellular eukaryotes and adhesives of infectious disease pathogens. The research will contribute to further understanding the potential mechanism of quorum-sensing molecules and the search for new anti-adhesive drugs that reduce the formation of biofilms. Keywords: adhesion, Aeromonas hydrophila, biofilm, Cyprinus carpio Linnaeus, optical density.
Research Article | 28 Oct 2022
Sangyod rice bran extract enhances Lacticaseibacillus paracasei growth during the exponential phase and antibacterial activity of L. paracasei supernatant against zoonotic and foodborne pathogens
Krittika Kabploy, Phirabhat Saengsawang, Chonticha Romyasamit, Suthinee Sangkanu, Warangkana Kitpipit, Thotsapol Thomrongsuwannakij, Tuempong Wongtawan, Mareena Daus, Maria de Lourdes Pereira, and Watcharapong Mitsuwan

Volume-15 | Issue-10 | Article-13 | https://doi.org/10.14202/vetworld.2022.2466-2474

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Prebiotics are a group of nutrients or compounds that are degraded by the gut microbiota, including Lacticaseibacillus paracasei. The probiotic plays an important role in adhesion to the gut and is able to produce antimicrobial substances to inhibit pathogens. This study aimed to investigate the effects of Sangyod rice bran extract on the growth promotion of L. paracasei. Furthermore, antibacterial activity of the extract and L. paracasei supernatants cultured in De Man, Rogosa and Sharpe (MRS) medium plus the extract against zoonotic and foodborne pathogens was investigated. Antibacterial activity of the crude extract and the oil from Sangyod rice bran against the pathogens, including Bacillus cereus, Staphylococcus aureus, Escherichia coli, Avian pathogenic E. coli, and Pseudomonas aeruginosa was investigated using broth microdilution assay. The effects of the crude extract and the oil on the growth and adhesion of L. paracasei were further determined. The antibacterial activity of L. paracasei supernatant cultured in the medium supplemented with the extract and the oil against the pathogens was determined by agar well diffusion assay, followed by the broth microdilution assay. Finally, the chemical constituents and antioxidant activity of the crude extract and the oil from Sangyod rice bran were investigated. The crude extract and the oil from Sangyod rice bran enhanced L. paracasei growth during the exponential phase. Furthermore, the crude extract at 0.25 mg/mL significantly enhanced the adhesion of L. paracasei to the surface compared with the control. Both minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values of the crude extract against B. cereus and S. aureus were 0.5 and 1.0 mg/mL, respectively. All pathogens were sensitive to the supernatant of L. paracasei with similar MIC and MBC ranging from 12.5% v/v to 50% v/v. However, the MIC and MBC values of L. paracasei supernatant grown in MRS medium plus the crude extract and oil were not significantly different compared to the supernatant obtained from MRS alone. The crude extract had free radical scavenging activities with IC50 values at 0.61 mg/mL. The results suggested the potential benefits of the crude extract from Sangyod rice bran for inducing the growth and the adhesion of L. paracasei and inhibiting zoonotic and foodborne pathogens. Keywords: Avian pathogenic Escherichia coli, Bacillus cereus, Lacticaseibacillus paracasei, Sangyod rice bran extract, Staphylococcus aureus.

Volume-15 | Issue-10 | Article-14 | https://doi.org/10.14202/vetworld.2022.2475-2490

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A delivery system consisting of bone marrow mesenchymal stem cells (MSCs) loaded with polyethylene glycol (PEG) coated superparamagnetic iron oxide nanoparticles (SPIONs) was constructed to treat a rat model of cisplatin (Cis)-induced nephrotoxicity with 1/10 of the common dose of anti-tumor necrosis factor-alpha (TNF-α) antibodies (infliximab). Morphology, size, crystallinity, molecular structure, and magnetic properties of uncoated and PEG-coated SPIONs were analyzed. A delivery system consisting of MSCs containing infliximab-labeled PEG-coated SPIONs (Infliximab-PEG-SPIONs-MSCs) was generated and optimized before treatment. Fifty female Wistar rats were divided into five equal groups: Group 1: Untreated control; Group 2 (Cis): Rats were administered Cis through intraperitoneal (i.p.) injection (8 mg/kg) once a week for 4 weeks; Group 3 (Infliximab): Rats were injected once with infliximab (5 mg/kg), i.p. 3 days before Cis administration; Group 4 (Cis + MSCs): Rats were injected with Cis followed by an injection of 2 × 106 MSCs into the tail vein twice at a 1-week interval; and Group 5 (Cis + Infliximab (500 μg/kg)-PEG-SPIONs-MSCs): Rats were injected with the delivery system into the tail vein twice at a 1-week interval. Besides histological examination of the kidney, the Doppler ultrasound scanner was used to scan the kidney with the Gray-color-spectral mode. In vivo, intra-renal iron uptake indicates the traffic of the delivery system from venous blood to renal tissues. Cis-induced nephrotoxicity resulted in a significant increase in TNF-α and malondialdehyde (MDA) (p < 0.05), bilirubin, creatinine, and uric acid (p < 0.01) levels compared with the untreated control group. The different treatments used in this study resulted in the amelioration of some renal parameters. However, TNF-α levels significantly decreased in Cis + Infliximab and Cis + MSCs (p < 0.05) groups. The serum levels of MDA significantly decreased in Cis + Infliximab (p < 0.05), Cis + MSCs (p < 0.05), and Cis + Infliximab-PEG-SPIONs-MSCs (p < 0.01). Furthermore, the serum activities of antioxidant enzymes were significantly elevated in the Cis + MSCs and Cis + Infliximab-PEG-SPIONs-MSCs groups (p < 0.05) compared to the Cis-induced nephrotoxicity rat model. With the support of the constructed MSCs-SPIONs infliximab delivery system, it will be possible to track and monitor cell homing after therapeutic application. This infliximab-loading system may help overcome some challenges regarding drug delivery to the target organ, optimize therapeutics' efficacy, and reduce the dose. The outcomes of the current study provide a better understanding of the potential of combining MSCs and antibodies-linked nanoparticles for the treatment of nephrotoxicity. However, further investigation is recommended using different types of other drugs. For new approaches development, we should evaluate whether existing toxicity analysis and risk evaluation strategies are reliable and enough for the variety and complexity of nanoparticles. Keywords: cisplatin, drug delivery system, infliximab, mesenchymal stem cells, nephrotoxicity, superparamagnetic iron oxide nanoparticles, tumor necrosis factor-alpha.
Research Article | 29 Oct 2022
Polymerase chain reaction-based detection of coinfecting DNA viruses in Vietnamese pigs in 2017 and 2021
Van Giap Nguyen, Huu Anh Dang, Thanh Trung Nguyen, Thi My Le Huynh, Ba Hien Nguyen, Le Anh Minh Pham, and Huynh Thanh Phuong Le

Volume-15 | Issue-10 | Article-15 | https://doi.org/10.14202/vetworld.2022.2491-2498

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Many studies have reported on the phenomenon of co-infections involving two or more pathogens (bacteria or viruses) over the past few years. However, very few studies on this issue were conducted in Vietnam. Therefore, this study aimed to determine the circulation of single and multiple porcine parvovirus (PPV) (e.g., PPV1, PPV2, PPV3, and PPV4), porcine bocavirus (PBoV), and torque teno virus (TTV) (TTV1 and TTV2) infections in Vietnamese pigs. A total of 174 porcine circovirus 2-positive samples from pigs (n = 86 for 2017 and n = 88 for 2021), including from the sera and internal organs, across 11 provinces were examined by polymerase chain reaction. This study demonstrated the wide distribution of DNA viruses among pig farms in Vietnam in 2021, with the detection rate for PPV ranging from 3.4% to 27.3% among PPV1-PPV4. Moreover, the detection rates of TTV genotypes were confirmed to be 14.8% (TTV1) and 63.6% (TTV2), respectively, and the positive rate of PBoV was 65.9%. The most frequent combinations were double and triple infections. Double infection was found in 16/86 (18.6%) in 2017 and 26/88 (29.5%) in 2021, while triple infection was found at 19/86 (22.1%) in 2017 and 26/88 (29.5%) in 2021. The incidence of simultaneous detection of more than three viruses was low. These results provide at least partial information about the occurrence of three viruses, including PPV (including PPV1 to 4), PBoV, and TTV (TTV1 and TTV2), in pigs. Determination of particular viruses in pigs will help to prevent the porcine respiratory disease complex caused by DNA viruses in Vietnamese pigs in the future. Keywords: co-infection, porcine bocavirus, porcine parvovirus, torque teno virus, Vietnamese pigs.
Research Article | 30 Oct 2022
Antibiotic resistance profiles and activity of clove essential oil (Syzygium aromaticum) against Pseudomonas aeruginosa isolated of canine otitis
Larissa Vieira Costa, Janaina Marcela Assunção Rosa Moreira, Isabela de Godoy Menezes, Valéria Dutra, and Arleana do Bom Parto Ferreira de Almeida

Volume-15 | Issue-10 | Article-16 | https://doi.org/10.14202/vetworld.2022.2499-2505

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Pseudomonas aeruginosa is often isolated from acute and chronic otitis and deep pyoderma in dogs. The increase in bacterial resistance to antibiotics induced the need for alternative therapies to treat infections, with an emphasis on essential oils (EOs). This study aimed to investigate clove oil's in vitro bactericidal action as a therapeutic alternative against strains of P. aeruginosa isolated from canine otitis. The antibacterial activity of clove oil was evaluated by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) using the broth microdilution technique in 96-well plates. Serial concentrations of 10–0.31% of the oil were used, equivalent to 104.5–3.26 mg/mL. The susceptibility of isolates to different classes of antibiotics was determined by the disk diffusion technique using 20 antibiotics belonging to eight classes. Isolates resistant to at least one antibiotic of three different classes were considered multidrug-resistant (MDR). A high occurrence of resistance was observed for three antibiotics belonging to the cephalosporin classes (cefadroxil, cephalexin, and ceftriaxone), namely, sulfamethoxazole + trimethoprime, doxycycline, and enrofloxacin. The lowest resistance rates were observed for meropenem (4.88%), amikacin (12.20%), and tobramycin (12.2%). All isolates were susceptible to clove oil with an equivalent MIC and MBC from 3.26 to 6.53 mg/mL. Eugenol was the major component of the oil. Clove EO was effective against MDR strains of P. aeruginosa, indicating an alternative for developing an efficient and low-cost antimicrobial agent to treat canine otitis. Keywords: essential oil, multidrug resistance, Pseudomonas aeruginosa, susceptibility.
Research Article | 31 Oct 2022
Molecular diagnosis of bovine tuberculosis on postmortem carcasses during routine meat inspection in Benin: GeneXpert® testing to improve diagnostic scheme
Cyrille K. Boko, Ange-Régis Zoclanclounon, Camus M. Adoligbe, Hebert Dedehouanou, Marguéritte M'Po, Samuel Mantip, and Souaïbou Farougou

Volume-15 | Issue-10 | Article-17 | https://doi.org/10.14202/vetworld.2022.2506-2510

Preview Abstract
Bovine tuberculosis (TB) is a zoonotic disease of major public health importance, particularly in African countries, where control measures are limited or largely not applied. This study aimed to determine the accuracy of the currently used bovine TB diagnostic method at slaughterhouses in Benin; this is to contribute to the betterment and improvement in the epidemiological surveillance of the disease in the country. A total of 40 tissue samples were collected from meat/viscera (lung, liver, heart, kidney, and the gastro-intestinal tract tissues) at Cotonou slaughterhouses from ruminants suspected to be infected with bovine TB during routine meat inspection. The collected samples were analyzed using GeneXpert testing technique as a reference method. Twenty-six samples tested positive out of the 40 suspected tissue samples collected by GeneXpert diagnostic technique; this shows the limitation of the routine meat inspection in detecting bovine TB as currently performed in Benin. The outcome of the use of the molecular technique, therefore, supports the importance of the use of a molecular tool alongside the routine meat inspection for a better understanding of the epidemiology of bovine TB in Benin. However, more robust technical and policy efforts are needed for a sustainable implementation of such a strategy. Keywords: genomic amplification, non-tuberculosis mycobacteria, one health, polymerase chain reaction accuracy, zoonotic tuberculosis.