In silico prediction of flavan-3-ol as a bioactive compound of Calophyllum macrophyllum as a potential drug against angiostrongylus eosinophilic meningitis

Research (Published online: 25-05-2022)

20. In silico prediction of flavan-3-ol as a bioactive compound of Calophyllum macrophyllum as a potential drug against angiostrongylus eosinophilic meningitis

Muhammad Hambal, Frengki Frengki, Wahyu Eka Sari and Henni Vanda

Veterinary World, 15(5): 1305-1313

ABSTRACT

Background and Aim: Angiostrongylus eosinophilic meningitis is caused by larvae of the rat lungworm Angiostrongylus cantonensis. It manifests as meningitis, radiculitis, cranial nerve abnormalities, and encephalitis, which can be fatal. A flavan-3-ol compound isolated from the bark of Calophyllum macrophyllum Scheff. has several medicinal properties, including antioxidant, anti-inflammatory, antidiabetic, and antibacterial activities. This compound is stronger than other types of flavan-3-ols such as catechin. This study aimed to identify the hydroxylation pattern of this flavan-3-ol compound and evaluated its potential as an anti-meningitis drug, using an in silico approach through pharmacophore and molecular docking methods.

Materials and Methods: Pharmacokinetic and toxicological data were analyzed and supported by the server http://www. swissadme.ch/index.php and https://tox-new.charite.de/protox_II/index.php. The hydroxylation pattern of the flavan-3-ol compound was identified using shear reagents (MeOH, NaOH, NaOAc, HCl, and AlCl3). The CviR receptor (pdb id.3QP5) was used in the in silico approach, and seven ligands were downloaded from PubChem in "SMILES" format.

Results: The spectroscopic analysis conducted using the shear reagents confirmed that the flavan-3-ol compound has a "p-diOH" pattern on the cinnamoyl ring. Pharmacophore analysis revealed this compound "hit" with pharmacophore features, and molecular docking analysis showed that this compound has a strong affinity with both receptors.

Conclusion: The flavan-3-ol compound is a potential drug candidate for meningitis caused by pathogenic bacteria and the worm A. cantonensis. This result was supported by the pharmacokinetic profile, which had a very low toxicity level to the host. However, further investigation is required to confirm the data in vitro and in vivo.

Keywords: Angiostrongylus cantonensis, Flavan-3-ol, molecular docking, pharmacophore.

Retail chicken giblets contaminated with extended-spectrum cephalosporin- and carbapenem-resistant Salmonella enterica carrying blaCMY-2

Research (Published online: 25-05-2022)

19. Retail chicken giblets contaminated with extended-spectrum cephalosporin- and carbapenem-resistant Salmonella enterica carrying blaCMY-2

Fatma Abdel-Kader, Eman Hamza, Khaled A. Abdel-Moein and Maha A. Sabry

Veterinary World, 15(5): 1297-1304

ABSTRACT

Background and Aim: Chickens are considered as the main source of Salmonella, with infection potentially spreading to the public through outlets. The study aimed to investigate poultry shops for Salmonella enterica resistant to extended-spectrum cephalosporins-resistant (ESCR) and carbapenems-resistant (CR).

Materials and Methods: Samples were collected from chicken giblets, water tanks, and workers at retail shops. Salmonella was isolated and serotyped; the presence of invA, stn, ompA, and ompF was determined using polymerase chain reaction (PCR). The isolates were tested for ESCR and CR by a disk-diffusion test; a confirmatory extended-spectrum β-lactamase (ESBL) test was performed by combinational disk-diffusion test with clavulanic acid. The resistant isolates were screened for ESBL (blaTEM, blaSHV, blaCTX-M, and blaOXA-1), AmpC blaCMY-2, and carbapenemase (blaKPC, blaNDM, and blaOXA-48) genes using PCR.

Results: S. enterica was isolated from chicken giblets (13/129) and the 13 isolates were ESCR. Based on the confirmatory ESBL test and CR, the 13 isolates were classified into the following resistance phenotypes: ESBL-producing and CR (n=4), ESBL-producing (n=1), non-ESBL-producing and CR (n=6), and non-ESBL-producing (n=2). All the five isolates with ESBL-producing phenotype carried predominantly blaTEM, blaSHV, and blaCMY-2. Regardless of being phenotypically CR, none of these isolates carried any of the tested carbapenemase genes. Surprisingly, the isolates with non-ESBL phenotype were found to carry blaTEM, blaSHV, and blaCMY-2. The blaKPC was present mainly in the isolates with non-ESBL and CR phenotypes. Interestingly, two isolates of the non-ESBL and CR phenotype showed resistance to cefepime, the fourth generation cephalosporins. Salmonella was also recovered from the water tanks (2/7) and the workers (2/16). The four isolates were ESCR and showed a non-ESBL-producing and CR phenotype; they harbored blaTEM, blaSHV, blaOXA-1, and blaKPC. The blaCMY-2 was found in one isolate from water and one from humans. All Salmonella isolates carried invA, stn, ompA, and ompF.

Conclusion: Virulent ESCR S. enterica were identified in retail shops. The isolates carried blaCMY-2 and ESBL-genes, with a high proportion showing CR. Transmission of such strains to humans through food leads us to recommend regular inspection of retail outlets for antibiotic-resistant bacteria.

Keywords: carbapenems, extended-spectrum β-lactamase, extended-spectrum cephalosporin-resistant, plasmid AmpC, Salmonella enterica.

Evaluation of antibody and antigen cross-reaction in Kenyan dairy cattle naturally infected with two pestiviruses: Bovine viral diarrhea virus and classical swine fever virus

Research (Published online: 24-05-2022)

18. Evaluation of antibody and antigen cross-reaction in Kenyan dairy cattle naturally infected with two pestiviruses: Bovine viral diarrhea virus and classical swine fever virus

Daniel Muasya, John Van Leeuwen, George Gitau, Shawn McKenna, Luke Heider and Joan Muraya

Veterinary World, 15(5): 1290-1296

ABSTRACT

Background and Aim: Bovine viral diarrhea virus (BVDV) and classical swine fever virus (CSFV) are important pathogens of cattle and pigs, respectively, and belong to the genus Pestivirus. As CSFV has been shown to infect cattle, it can create diagnostic challenges of BVDV results through possible cross-reactivity where cattle could be exposed to pigs and CSFV. This study aimed to determine the possible cross-reactivity of BVDV and CSFV enzyme-linked immunosorbent assay (ELISA) results for antigen (Ag) and antibody (Ab) among smallholder dairy cattle in Kenya.

Materials and Methods: This was a cross-sectional study based on a single visit to farms to collect serum samples and other descriptive farm-level and animal-level information. Testing for BVDV Ag and Ab was conducted on serum samples from 320 dairy cows and heifers, with CSFV Ag and Ab testing conducted on a subset of 133 and 74 serum samples, respectively. CSFV testing was based on BVDV test results and the availability of enough sample volume from farms that kept pigs. The Ag and Ab tests utilized IDEXX ELISA for both BVDV and CSFV.

Results: For the 74 samples with Ab tests for both viruses, 40 (54.0%) were BVDV Ab positive, while 63 (85.1%) were CSFV Ab positive. Of the 40 BVDV Ab positive samples, 36 cattle (90.0%) tested positive for CSFV Ab. However, of the 34 BVDV Ab negative samples, 27 (79.4%) were CSFV Ab test-positive. For the 133 samples with Ag tests for both viruses, 125 (94.0%) were BVDV Ag positive, while 2 (1.5%) samples were CSFV Ag positive. None of the eight BVDV Ag negative samples was positive for CSFV Ag and only two (1.6%) of the 125 BVDV Ag positive samples were positive for CSFV Ag.

Conclusion: The results indicate either substantial cross-reactivity of the two Ab ELISA tests, or reactivity with some other protein in the samples that led to the positive Ab test results. There was only limited evidence for cross-reactivity of the two Ag ELISA tests. We recommend that Pestivirus genus cross-reactivity be considered when interpreting BVDV ELISA results in cattle, more for Ab than Ag tests. Further research is needed to clarify the levels of cross-reactivity between BVDV and other Pestivirus Ag and Ab tests from animals on mixed-species farms.

Keywords: antibody, antigen, bovine viral diarrhea virus, classical swine fever virus, cross-reactivity, smallholder dairy.

Crude extract of Jatobá leaves promotes canine osteosarcoma cell D17 proliferation

Research (Published online: 24-05-2022)

17. Crude extract of Jatobá leaves promotes canine osteosarcoma cell D17 proliferation

V. S. Vieira, V. S. Cruz, L. L. Nepomuceno, N. P. Soares, E. Arnhold, W. F. P. Teixeira, D. S. Vieira, J. C. A. Borges, F. M. Paixão and E. G. Araújo

Veterinary World, 15(5): 1283-1289

ABSTRACT

Background and Aim: New substances for neoplasm treatment have to be carefully studied to minimize adverse effects and prevent disease progression stimulation. Jatobá is a typical tree of the Cerrado and Caatinga biome, with antifungal, antimicrobial, larvicide, antioxidant, and antiproliferative properties. This study aimed to investigate the action of the crude extract of Jatobá leaves (EBFJ) on canine osteosarcoma (CO) cells and analyze the expression of biomarkers in neoplasm progression.

Materials and Methods: D17 cells were cultured and subjected to treatment with EBFJ at different concentrations (10 μg/ mL; 100 μg/mL; 1000 μg/mL; 2000 μg/mL; and 5000 μg/mL) and exposure times (24 h, 48 h, and 72 h). The tetrazolium reduction assay and the immunocytochemistry technique, with anti-Bcl2, anti-p53, and anti-Ki-67 antibodies, were used to observe the effect of the extract on cell proliferation.

Results: Doses of 2000 μg and 5000 μg had cell viability of 300.80% and 361.84%, respectively. The extract did not show significant cytotoxicity of samples with the control group. The confluence of cells, the number of labeled cells, and the expression of Bcl2, Ki-67, and p53 were higher in the groups treated with EBFJ, with a statistical difference from the group without treatment.

Conclusion: EBFJ was not cytotoxic and had a proliferative effect on CO D17 cells. The confluence of cells, the number of labeled cells, and the expression of Bcl2, Ki-67, and p53 were higher in the groups treated with the extract.

Keywords: antineoplastic chemotherapy, biodiversity, bone tumors, Hymenaea martiana Hayne, medicinal plants.

In vivo test of Vibrio alginolyticus and Vibrio harveyi infection in the humpback grouper (Cromileptes altivelis) from East Java Indonesia

Research (Published online: 23-05-2022)

16. In vivo test of Vibrio alginolyticus and Vibrio harveyi infection in the humpback grouper (Cromileptes altivelis) from East Java Indonesia

Uun Yanuhar, Hendra Nurcahyo, Luluk Widiyanti, Nur Sakinah Junirahma, Nico Rahman Caesar and Sukoso Sukoso

Veterinary World, 15(5): 1269-1282

ABSTRACT

Background and Aim: The need for fish seeds resistant to bacterial and viral infections has encouraged studies on the molecular pathogenesis mechanism of Vibrio bacteria, such as Vibrio alginolyticus and Vibrio harveyi, regarding the receptor organs, protein adhesion mechanisms, and antibody responses of the humpback grouper. This study aims to confirm the characteristics of the specific proteins expressed in the receptor organ of the humpback grouper (Cromileptes altivelis) using the expression of V. alginolyticus and V. harveyi bacteria.

Materials and Methods: The study was conducted by isolating crude protein and whole cells from both the Vibrio bacteria. In addition, serum and organ tissue were also isolated from fish samples. Then, hemagglutination and dot blot tests with polyacrylamide gel electrophoresis analysis were performed to determine the highest expression of receptor from the whole bacterial cells and crude protein from both healthy and infected (V. alginolyticus and V. harveyi) fishes. Scanning electron microscope results showed that V. alginolyticus and V. harveyi could express bundle-forming pili, which is involved in bacterial autoaggregation and the mediation of the initial attachment of bacteria to their host cells.

Results: These results indicated that all the specific receptors for protein in fish organs recognized vibriosis antigens. The specificity test showed that the brain, eye, and kidney organs' receptors provided a quality and quantity level of responses at 22.63, 53.95, and 43.15 kDa, respectively. The polyclonal anti-V. alginolyticus immunoglobulin M (IgM) antibodies were more cross-reactive than the anti-V. harveyi IgM. Hence, this shows that V. alginolyticus bacteria are more pathogenic than V. harveyi.

Conclusion: In the future, the molecular characteristics of V. alginolyticus and V. harveyi antigens and the specific receptor organ proteins in the humpback grouper can be developed as the basis for constructing molecular peptide-based vaccine materials.

Keywords: crude protein, humpback grouper, Vibrio alginolyticus, Vibrio harveyi, western blot.

Evaluation of room temperature (30°C to 35°C) lyophilized vaccine with radio inactivated Mannheimia haemolytica whole cells isolated from infected sheep

Research (Published online: 23-05-2022)

15. Evaluation of room temperature (30°C to 35°C) lyophilized vaccine with radio inactivated Mannheimia haemolytica whole cells isolated from infected sheep

Sahar Ahmed, Waleed Abdelgaber Nemr, Walaa Awad Ahmed Mohamed, Amany Mohamed Mohamed and Mohamed Abd El-Fatah Mahmoud

Veterinary World, 15(5): 1261-1268

ABSTRACT

Background and Aim: Vaccines are one of the important tools for fighting diseases and limiting their spread. The development of vaccines with high efficacy against diseases is essential. Ionizing radiation is the method used for the preparation of the irradiated gamma Mannheimia haemolytica vaccine. The study aimed to measure the metabolic activity and electron microscopic examination of the irradiated bacterial cells and immunological efficiency of different preparations of the irradiated M. haemolytica vaccine.

Materials and Methods: The irradiated vaccines were prepared in three forms at a dose of 2×109 colony-forming unit (CFU) (irradiated M. haemolytica, trehalose irradiated M. haemolytica, and trehalose lyophilized irradiated M. haemolytica). The formalin-killed vaccine was prepared at a dose of 2×109 CFU. Scanning electron microscopy was used to determine the difference between the non-irradiated bacterial cells and the bacterial cells exposed to gamma radiation. The metabolic activity of the irradiated bacterial cells was measured using the Alamar blue technique. Rabbits were divided into five groups (control, vaccinated groups with the formalin-killed vaccine, irradiated bacterial cells without trehalose, trehalose irradiated bacteria, and trehalose lyophilized irradiated bacterial cells). The rabbits were subcutaneously inoculated twice in 2-week intervals. Enzyme-linked immunosorbent assay, interferon-gamma (IFNγ), and interleukin 4 (IL4) assays were used to evaluate the vaccines' immunological efficiency in rabbits.

Results: The metabolic activity tests showed that the bacterial cells exposed to gamma radiation at the lowest lethal dose have metabolic activity. The difference in the metabolic activity between preparations of the irradiated bacterial cells varied according to the cell concentration and incubation time. The highest level of metabolic activity was 8 h after incubation in the nutrient broth medium compared with 4 and 18 h. The scanning electron microscopy of irradiated bacterial cells showed a cavity at the bacterial cell center without rupture of the surrounding cell membrane compared to the non-irradiated bacterial cells. The antibody level in the groups vaccinated with the different preparations of the irradiated bacterial cells was high compared with the control and formalin-killed vaccine groups. The level of the IFNγ showed an increase after the second dose in the group vaccinated with irradiated bacterial cells without trehalose compared with the other groups. The IL4 level in the vaccinated groups with the irradiated bacterial cells without trehalose, irradiated bacterial cells with trehalose, and trehalose lyophilized irradiated bacterial cells were at a high level when compared with the formalin-killed vaccinated group and control group after the second inoculation.

Conclusion: The irradiated M. haemolytica vaccine provides a wide range of humoral and cellular immunity. This study showed high immunological efficiency in rabbits inoculated with the irradiated M. haemolytica vaccine that was shown in the high levels of antibodies (IFNγ and IL4) compared with the group treated with the formalin-killed vaccine. The second dose of irradiated M. haemolytica vaccine is an immune booster that gives the irradiated vaccine a long-acting immunological efficiency.

Keywords: enzyme-linked immunosorbent assay, gamma radiation, interferon-gamma, interleukin 4, Mannheimia haemolytica, vaccine.

Prevalence of susceptibility to Cryptosporidium spp. among dairy calves with different feeding regimens with an emphasis on the feeding of transition milk

Research (Published online: 22-05-2022)

14. Prevalence of susceptibility to Cryptosporidium spp. among dairy calves with different feeding regimens with an emphasis on the feeding of transition milk

Alīna Zolova, Dace Keidāne and Maksims Zolovs

Veterinary World, 15(5): 1256-1260

ABSTRACT

Background and Aim: Colostrum composition and importance for newborn organisms were repeatedly studied. However, the interest in transitional milk usefulness is weak and recommendations concerning transition milk intake are not developed. The aim of this study was to evaluate whether transition milk intake after colostrum consumption affects the chances of calf infection with Cryptosporidium spp.

Materials and Methods: We collected data for Cryptosporidium spp. infection from calves (n=425) divided into three groups: The first group – supervised colostrum and transition milk intake; the second group – supervised colostrum and whole milk intake; and the third group – not supervised colostrum and whole milk intake. To detect oocysts of Cryptosporidium spp. in feces, the flotation method was used, and slides were stained using the modified Ziehl-Neelsen method. Generalized linear mixed modeling was conducted to determine whether the explanatory variable – the management of colostrum and transition milk feeding with three categories (three research groups) – was related to the probability of calves incurring infection with Cryptosporidium spp.

Results: In the first group, 26.1% of calves were positive for the presence of Cryptosporidium spp. oocysts, in the second – 37.2%, and in the third – 44.1%. Statistical data analysis showed that calves who did not receive transition milk after colostrum consumption had increased chances of having Cryptosporidium spp. (by 1.90-2.47 times on average). The main results showed that the management of colostrum and transition milk feeding is related to Cryptosporidium spp. infection, indicating that both colostrum and transitional milk play a significant role in controlling pathogenic infections.

Conclusion: The most effective management of colostrum and transition milk feeding against Cryptosporidium spp. infection is the timely intake of an adequate amount of colostrum followed by transitional milk consumption for at least 2 weeks before weaning from the dam.

Keywords: calves, colostrum, Cryptosporidium, neonatal diarrhea, transition milk.