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.

Semen characteristics and second successful artificial insemination of Asian elephant (Elephas maximus) in Thailand

Research (Published online: 22-05-2022)

13. Semen characteristics and second successful artificial insemination of Asian elephant (Elephas maximus) in Thailand

Ampika Thongphakdee, Supalak Kiatsomboon, Sakhon Noimoon, Urarikha Kongprom, Itti Boonorana, Santhita Karoon, Jedsada Thawnern, Apichaya Sakulthai, Petthisak Sombutputorn, Manakorn Sukmak, Chainarong Punkong and Nikorn Thongtip

Veterinary World, 15(5): 1246-1255

ABSTRACT

Background and Aim: As the number of wild Asian elephants (Elephas maximus) continues to decline, maintaining healthy populations under human care is vital. Male fertility assessment is essential for understanding the reproductive status, which can help to uncover underlying problems and improve the rate of pregnancy success. The objectives of this study in Asian elephants were as follows: (1) To investigate the semen characteristics; (2) to compare the relative seminal vesicle size and semen characteristics; (3) to compare the semen characteristics between good-motile (>60% progressive motility) and poor-motile (<60% progressive motility) ejaculates; and (4) to investigate the pregnancy success rate after artificial insemination (AI) with combined chilled and frozen semen.

Materials and Methods: In total, 153 ejaculates were collected by manual rectal stimulation from 25 bulls. The volume, pH, sperm concentration, progressive motility, viability, morphology, and membrane integrity were investigated in each ejaculate. Assessment of accessory sex glands was conducted using transrectal ultrasonography to compare the relative seminal vesicle size and semen characteristics, and the bulls were divided into two groups according to the size of the ampulla (<7 or ≥7 cm2). For the comparison of good and poor-motile ejaculates and semen characteristics, the samples were divided into two groups: Good-motile (>60% progressive motility) and poor-motile (<60% progressive motility) ejaculates. Semen ejaculates for AI were collected from three bulls. The estrous cycles of four females were monitored using an enzyme immunoassay. Seven AI attempts were conducted using frozen and/or chilled semen by endoscopic visualization. AI was repeated 1 day before the luteinizing hormone surge, on the day of the surge, and 1 day after the surge. Pregnancy was confirmed by monitoring the serum progesterone profile and the abdomen and mammary glands changes.

Results: From 153 ejaculates, the mean±standard error values of progressive motility, semen volume, sperm concentration, pH, and viability were 40.18%±2.28%, 40.94±3.86 mL, 1,205.58±62.26×106 sperm/mL, 7.50±0.10, and 56.17%±1.96%, respectively. Comparing ampulla size and semen characteristics revealed that the bulls with ampullae of ≥7 cm2 yielded significantly larger volume ejaculates. However, there were no significant differences in sperm motility and concentration. The comparison of semen characteristics between good- and poor-motile ejaculates revealed that the former had significantly higher pH, viability, normal acrosomes, intact membranes, and normal head and tail morphology but often had a significantly lower volume and sperm concentration. From seven AI attempts in four females, one female had a confirmed pregnancy (14.3% pregnancy rate), and delivered a healthy live female baby weighing 128 kg at 21 months and 12 days of gestation. The baby is now 3 years old and in a healthy condition, with normally developing growth and behavior.

Conclusion: The semen characteristics of Asian elephants can be used as the baseline reference for further applications. The ampullae size indicates semen quantity but not quality. Our success in producing an elephant calf from AI using frozen and chilled semen demonstrated that AI can be used as an alternative approach for the breeding management of Asian elephants. However, the semen of Asian elephants is of poor quality, especially in terms of membrane integrity; thus, the improvement in semen quality through intensive and careful management of elephant health and fertility remains a challenge for the future. Furthermore, a sperm bank should be established to develop sperm cryopreservation, which will be invaluable for improving the genetic diversity of the Asian elephant.

Keywords: artificial insemination, Asian elephant, hormone, semen quality, sex accessory glands.

Designing a rabies control mobile application for a community-based rabies surveillance system during the COVID-19 pandemic in Bali, Indonesia

Research (Published online: 21-05-2022)

12. Designing a rabies control mobile application for a community-based rabies surveillance system during the COVID-19 pandemic in Bali, Indonesia

I Made Subrata, Ngakan Putu Anom Harjana, Kadek Karang Agustina, Sang Gede Purnama and Made Pasek Kardiwinata

Veterinary World, 15(5): 1237-1245

ABSTRACT

Background and Aim: Rabies remains a public health concern in Indonesia, and the coronavirus disease (COVID-19) pandemic has stymied rabies prevention and control efforts. There is a need to transform the rabies program to be adaptable to pandemic situations to improve program coverage on dog vaccination and rabies surveillance. This study aimed to create a rabies control (RaCon) mobile application for a community-based rabies surveillance system during COVID-19 in Bali, Indonesia.

Materials and Methods: We employ the Design Science Research methodology. Surveillance officers, veterinarians, community leaders, outreach workers, and dog owners participated in a series of offline in-depth interviews and focus group discussions. The RaCon prototype was evaluated using the Post-Study System Usability Questionnaire (PSSUQ) framework, which included the system's usefulness, information quality, and interface quality. In this study, we used both a qualitative (n=50) and quantitative (n=342) approach.

Results: According to the findings of this study, integrating public health and animal health into the rabies surveillance system are critical to supporting the One Health approach and encouraging community engagement in rabies programs. The RaCon prototype is expected to include features such as pet ownership, case report, news and announcements, nearest vet, health information, outbreak radar, emergency call, and app feedback. The RaCon prototype passed both qualitative and quantitative evaluations, indicating that it could be used to support the rabies surveillance system, particularly in the COVID-19 situation.

Conclusion: The RaCon prototype was accepted by the users and got positive feedback in terms of the system's usefulness, information quality, and interface quality dimension. As a result, this prototype has the potential to be integrated into the rabies surveillance system in Bali, particularly to strengthen the community-based rabies surveillance system. Even though this prototype received positive feedback, this study focuses solely on the design development and evaluation of its user interface. As a result, further development is required before incorporating RaCon into the rabies prevention and control program.

Keywords: community-based intervention, One Health, post-study system usability questionnaire, rabies, surveillance system.

Vermikom feed additive effects on dairy cows' blood and milk parameters

Research (Published online: 21-05-2022)

11. Vermikom feed additive effects on dairy cows' blood and milk parameters

S. K. Sherimova, N. B. Sarsembayeva, T. B. Abdigaliyeva and B. Lozowicka

Veterinary World, 15(5): 1228-1236

ABSTRACT

Background and Aim: Dairy cattle breeding plays a significant role in providing the population with high-quality, reasonably priced goods. The development of this industry and its effectiveness depends on the proper use of available feed products. Feed additives (FAs), as a rule, should compensate for missing elements in the diet. This study aimed to determine the effect of the FA Vermikom on blood parameters, as well as milk physicochemical and mineral composition and yield in lactating dairy cows.

Materials and Methods: A total of 30 Holstein cows, with an average weight of 650±5 kg, were randomly divided into three groups of 10. Over a period of 5 months, each group was fed one of three diets: Mixed main diet without supplements (control), main diet supplemented with 2% Vermikom, and main diet supplemented with 4% Vermikom.

Results: Hematological parameters, productivity, and physicochemical and mineral composition of milk from animals provided the FA Vermikom were higher than those of the control group. In the Vermikom groups, the hemoglobin content was higher by 5.75%, calcium by an average of 10.8%, and total protein by 2.5%. The average daily milk yield was also higher by 8.4% than the control group. Regarding the mineral composition of the milk, the content of calcium, phosphorus, and iron exceeded that of the control on average by 2.9%, 3.4%, and 14.8%, respectively.

Conclusion: We propose introducing 4% Vermikom into the diet of dairy cows based on the study results. Future research will expand our knowledge regarding cows' needs for all recommended nutrients, thereby improving animal productivity and milk quality. The results obtained contribute to further expanding the food base of animal husbandry in the Republic of Kazakhstan.

Keywords: diet, feed additive, hematology, milk productivity, milk quality, vermiculite.