Expression of recombinant Omp18 and MOMP of Campylobacter jejuni and the determination of their suitability as antigens for serological diagnosis of campylobacteriosis in animals

Research (Published online: 30-01-2023)

27. Expression of recombinant Omp18 and MOMP of Campylobacter jejuni and the determination of their suitability as antigens for serological diagnosis of campylobacteriosis in animals

Sergey Borovikov, Kanat Tursunov, Alfiya Syzdykova, Ainagul Begenova, and Alfira Zhakhina

Veterinary World, 16(1): 222-228

ABSTRACT

Background and Aim: Campylobacteriosis causes gastrointestinal tract lesions in adults and children and may result in severe complications. The primary sources of infection are infected animals and animal products. Immunochemical methods effectively diagnose intestinal infections but require highly specific antigens to detect their antibodies. This study aimed to obtain two recombinant immunogenic antigens of Campylobacter jejuni, an outer membrane protein with a molecular weight of 18 kDa (Omp18) and the major outer membrane protein (MOMP) with a molecular weight of 45 kDa, and evaluate their suitability for the serological diagnosis of campylobacteriosis using immunochromatographic assay (ICA).

Materials and Methods: The C. jejuni Omp18 and MOMP gene sequences were synthesized de novo (Macrogen, Korea) and cloned into the pET32 expression plasmid. Using these genetic constructs, electrocompetent cells of the Escherichia coli BL21 strain were transformed and cultured under various conditions. Antigens were purified and refolded using metal affinity chromatography. The properties of the purified proteins were studied by western blotting, liquid chromatography with tandem mass spectrometry, and enzyme-linked immunosorbent assay (ELISA).

Results: We developed two recombinant E. coli BL21 cells producing rOmp18 and Recombinant MOMP (rMOMP) antigens with molecular weights of 36 and 64 kDa, respectively. Amino acid sequence analysis of the obtained antigens showed complete homology with the reference sequences in the PubMed NCBI database. Western blotting using positive-control sera demonstrated the specificity of the recombinant antigens. The results of ELISA with 94 bovine sera showed the interaction of recombinant antigens with specific antibodies.

Conclusion: The obtained rOmp18 and rMOMP antigens can detect antibodies in the serum of infected or recovered animals and can be used to develop ICA.

Keywords: Campylobacter jejuni, campylobacteriosis, diagnostics, outer membrane proteins, recombinant antigens.

Supplementation of Water Spinach (Ipomoea aquatica) on the utilization of Mimosa pigra and Leucaena leucocephala leaf for in vitro fermentation

Research (Published online: 30-01-2023)

26. Supplementation of Water Spinach (Ipomoea aquatica) on the utilization of Mimosa pigra and Leucaena leucocephala leaf for in vitro fermentation

Channy Sambo, Sreychou Heng, Pisey Vong, Kuyhor Te, Sath Keo, Mom Seng, and Samnang Ven

Veterinary World, 16(1): 215-221

ABSTRACT

Background and Aim: Ipomoea aquatica (Water Spinach) is the most potential for livestock growth performance, including chickens, pigs, cattle, and goats, especially in a tropical country like Cambodia. It is not only an alternative feed source but also one kind of supplemented feed for goat raising. Supplementation with Water Spinach in the utilization of low-quality tree foliage results in an increase in dry matter intake in goat production. This study aimed to identify the effectiveness of supplementation of Water Spinach in the utilization of Mimosa pigra and Leucaena leucocephala leaf in in vitro fermentation.

Materials and Methods: The study was designed according to a 2 × 2 factorial arrangement in randomized design of seven treatments with different ratios consisted of different three types of dietary treatments, including M. pigra, L. leucocephala, and Water Spinach. The treatments were arranged according to a completely randomized design and were as follow: T1 = M. pigra leaf (100%); T2 = L. leucocephala leaf (100%); T3 = M. pigra leaf and L. leucocephala leaf (50% and 50%); T4 = M. pigra leaf and Water Spinach (99.5% and 0.5%); T5 = L. leucocephala leaf and Water Spinach (99.5% and 0.5%); T6 = M. pigra leaf and Water Spinach (99% and 1%); and T7 = L. leucocephala leaf and Water Spinach (99% and 1%). A total of 200 mg (dry matter) of dietary treatments were prepared in a 60 mL syringe. Each treatment was replicated 3 time. Gas recording of each treatment lasted for 3 days. In vitro was performed for 72 h, was followed by Makkar method. Gas production was recorded at 2, 4, 8, 12, 24, 36, 48, and 72 h of incubation by using strict anaerobic technique. A mixture of rumen fluid and dietary treatments were carried out under continuous flushing with CO2 in sharking incubator at 39°C. After incubating for 72 h, the ammonia concentration (NH3-N) was measured and recorded to identify pH, nutrient digestibility, and ammonia concentration (NH3-N).

Results: Nutrient digestibility of the treatment with Water Spinach supplement in the utilization of L. leucocephala was obtained at a higher digestibility than treatment with M. pigra (p < 0.05). Gas production was different between groups (p < 0.05). Treatment with only M. pigra leaf had the highest gas production (A), while treatment with Water Spinach supplementation had the lowest gas production (A). At 0–24 h, the treatment with L. leucocephala leaf and Water Spinach 0.5% had the highest gas production, but after 24 h, M. pigra leaf and Water Spinach 1% and L. leucocephala leaf and Water Spinach 0.5% produced more gas compared to the other treatments (p < 0.05).

Conclusion: The supplementation of Water Spinach 1% in treatment with M. pigra and L. leucocephala leaf resulted in increased degradability, gas production, and NH3-N concentration without a change in the pH value rumen condition. Based on these results, it is recommended that the level of Water Spinach supplementation should be 1% of dietary intake. Future studies should consider investigating the rumen ecology associated with Water Spinach supplementation. Feeding with Water Spinach remains a good supplement for ruminant performance; therefore, further studies should be conducted using Water Spinach in ruminant feeding in both metabolic and feeding trials.

Keywords: In vitro fermentation, Leucaena leucocephala leaf, Mimosa pigra, Nutrient digestibility, Supplementation, Utilization, Water Spinach (Ipomoea Aquatica).

A novel portable Raman scattering platform for antibiotic screening in pig urine

Research (Published online: 29-01-2023)

25. A novel portable Raman scattering platform for antibiotic screening in pig urine

Nungnit Wattanavichean, On-uma Nimittrakoolchai, Noppadon Nuntawong, Mati Horprathum, Pitak Eiamchai, Saksorn Limwichean, Pacharamon Somboonsaksri, Donruethai Sreta, and Sirilak Meesuwan

Veterinary World, 16(1): 204-214

ABSTRACT

Background and Aim: Public health and food safety are gaining attention globally. Consumer health can be protected from chemical residues in meat by early detection or screening for antibiotic residues before selling the meat commercially. However, conventional practices are normally applied after slaughtering, which leads to massive business losses. This study aimed to use portable surface-enhanced Raman spectroscopy (SERS) equipped with multivariate curve resolution-alternation least squares (MCR-ALS) to determine the concentrations of enrofloxacin, oxytetracycline, and neomycin concentrations. This approach can overcome the problems of business loss, costs, and time-consumption, and limit of detection (LOD).

Materials and Methods: Aqueous solutions of three standard antibiotics (enrofloxacin, oxytetracycline, and neomycin) with different concentrations were prepared, and the LOD for each antibiotic solution was determined using SERS. Extracted pig urine was spiked with enrofloxacin at concentrations of 10, 20, 50, 100, and 10,000 ppm. These solutions were investigated using SERS and MCR-ALS analysis. Urine samples from pigs at 1 and 7 days after enrofloxacin administration were collected and investigated using SERS and MCR-ALS to differentiate the urinary enrofloxacin concentrations.

Results: The LOD of enrofloxacin, oxytetracycline, and neomycin in aqueous solutions were 0.5, 2.0, and 100 ppm, respectively. Analysis of enrofloxacin spiking in pig urine samples demonstrated the different concentrations of enrofloxacin at 10, 20, 50, 100, and 10,000 ppm. The LOD of spiking enrofloxacin was 10 ppm, which was 10 times lower than the regulated value. This technique was validated for the first time using urine collected on days 1 and 7 after enrofloxacin administration. The results revealed a higher concentration of enrofloxacin on day 7 than on day 1 due to consecutive administrations. The observed concentration of enrofloxacin was closely correlated with its circulation time and metabolism in pigs.

Conclusion: A combination of SERS sensing platform and MCR-ALS is a promising technique for on-farming screening. This platform can increase the efficiency of antibiotic detection in pig urine at lower costs and time. Expansion and fine adjustments of the Raman dataset may be required for individual farms to achieve higher sensitivity.

Keywords: antibiotics, pig, Raman spectroscopy, urine.

Influence of age at slaughter and sex on carcass characteristics and technological and sensory quality of Goliath chicken meat

Research (Published online: 28-01-2023)

24. Influence of age at slaughter and sex on carcass characteristics and technological and sensory quality of Goliath chicken meat

A. G. A. Christie Ahokpossi, A. Gabriel Bonou, Kokou Tona, Issaka Youssao Abdou Karim, and Yaovi Ameyapoh

Veterinary World, 16(1): 194-203

ABSTRACT

Background and Aim: The Goliath chicken is a slow-growing chicken with a high slaughter weight but whose carcass characteristics and meat quality have not yet been documented. This study aimed to evaluate the effect of age at slaughter and sex on the carcass characteristics and technological and sensory meat quality of Goliath chickens raised in southern Benin.

Materials and Methods: Data on the carcass characteristics and technological and sensory quality of meat were collected from 80 chickens raised in confinement and divided into two groups. The first group was reared for 12 weeks of age and the second group was for 20 weeks of age. The animals were individually weighed using an electronic balance (Terraillon 5000g) and then slaughtered. The different parts of the chicken carcass was weighed using the same method.

Results: The live, hot carcass, cold carcass, wing, thigh, and tail weights of males were significantly greater than those of females (p < 0.01). The wishbone muscle and thigh and tail assembly pH was lower in the 20-week-old chickens than in the 12-week-old chickens. The brightness and red index of the wishbone of the 20-week-old chickens were significantly higher than those of the 12-week-old chickens (p < 0.001). The red index of the wishbone and the yellow index of the thigh and tail assembly of females were lower than those of males.

Conclusion: The meat of 20-week-old Goliath chickens was juicier than that of 12-week-old chickens. Thus, the carcass composition of Goliath chickens is better at 20 weeks of age, especially in males.

Keywords: Benin, carcass, goliath chicken, meat, quality.

Antiprotozoal activity of Boesenbergia rotunda (L.) Mansf and Ganoderma lucidum (Fr.) Kart extracts against Blastocystis hominis

Research (Published online: 28-01-2023)

23. Antiprotozoal activity of Boesenbergia rotunda (L.) Mansf and Ganoderma lucidum (Fr.) Kart extracts against Blastocystis hominis

Chalermpon Kaewjai, Aunchalee Tonsomboon, Jaturawat Pawiwongchai, and Oranan Prommano

Veterinary World, 16(1): 187-193

ABSTRACT

Background and Aim: Blastocystis hominis is an intestinal protozoan in humans and animals. The parasite causes mild-to-severe intestinal complications, such as diarrhea, in healthy humans and immunocompromised hosts. This study aimed to determine the antiprotozoal activity of Boesenbergia rotunda (L.) Mansf and Ganoderma lucidum (Fr.) Kart extracts against B. hominis.

Materials and Methods: Antiprotozoal activity of B. rotunda and G. lucidum extracts against B. hominis subtype 3 was determined using the erythrosin B exclusion assay, confirmed by a time-kill study. The morphology of the parasite treated with the extracts was observed by a scanning electron microscope. The phytochemicals present in B. rotunda and G. lucidum extracts were identified by gas chromatography-mass spectrometry analysis.

Results: Both B. rotunda and G. lucidum extracts demonstrated strong antiprotozoal activity with similar minimum inhibitory concentration (MIC) values of 62.5 μg/mL. At 4× MIC and 8× MIC, both B. rotunda and G. lucidum extracts, and metronidazole inhibited the growth of B. hominis by up to 90% after 12 h treatment. Blastocystis hominis cells treated with B. rotunda extract, G. lucidum extract, and metronidazole were deformed and withered when compared with the control. Geraniol and versalide were found as the main compounds in B. rotunda and G. lucidum extracts, respectively.

Conclusion: These results indicate the potential medicinal benefits of B. rotunda and G. lucidum extracts in the growth inhibition of B. hominis.

Keywords: antiprotozoal activity, Blastocystis hominis, Boesenbergia rotunda, Ganoderma lucidum.

Chicken Enterococcus faecalis-induced immunoglobulin Y as a prophylactic and therapeutic agent against streptococcosis in red tilapia (Oreochromis hybrid)

Research (Published online: 28-01-2023)

22. Chicken Enterococcus faecalis-induced immunoglobulin Y as a prophylactic and therapeutic agent against streptococcosis in red tilapia (Oreochromis hybrid)

Rifky Rizkiantino, Fachriyan Hasmi Pasaribu, Retno Damajanti Soejoedono, Wyanda Arnafia, Dinda Reisinta, Rifaldi Iqbal Yadiansyah, Beni Halalludin, Yunita Ardini, Granita Khanaria, and I Wayan Teguh Wibawan

Veterinary World, 16(1): 175-186

ABSTRACT

Background and Aim: Streptococcosis is a common bacterial disease in red tilapia, in which Enterococcus faecalis infection has not been widely reported. This study aimed to evaluate the efficacy of pellets that contain chicken E. faecalis-induced immunoglobulin Y (IgY) to treat and prevent streptococcosis in red tilapia.

Materials and Methods: We conducted a 28-day study for immunoprophylaxis and immunotherapy, each using four groups with two replications: Healthy control fish (KS), non-IgY pellets (PA and TA), pellets with 25% egg yolk containing E. faecalis-induced IgY (PB and TB), and pellets with 50% egg yolk containing E. faecalis-induced IgY(PC and TC). Indirect enzyme-linked immunosorbent assay was performed on prototype pellets produced with an IgY suspension at 1.63 mg/mL as the standard optical density curve. For the immunoprophylaxis study, pellets of 3% of the average body weight of the experimental fish (0.50 g per fish per day) were given daily until day 14 before the challenge test with E. faecalis (2.1 × 109 Colony-forming unit/mL peroral) on day 15. The data from the observation period on days 15–28 were analyzed. For the immunotherapy study, pellets of 3% of the average body weight (0.50 g per fish per day) were given daily for 21 days (days 8–28) 7 day spost-infection. The data from the immunotherapy study were collected during the observation period on days 8–28. Statistical analysis was performed on non-specific immune variables: Total leukocytes, monocytes, lymphocytes, neutrophils, phagocytic activity, and macrophage capacity; and the semi-quantitative distribution of melanomacrophage centers (MMCs) in the lymphoid organs, such as spleen and liver. Photomacrographic data were analyzed descriptively and qualitatively by comparing the healing process and clinical signs found between experiments in the immunotherapy study.

Results: The pellet with 50% egg yolk with an IgY at 2.43 mg/g pellet, 3% of body weight once daily, was the best formula on experimental fish. The administration of this formulation can also increase non-specific immunity and the distribution of MMCs in the spleen and liver with a survival rate of 55% for 14 days of challenge period in the immunoprophylaxis study and 70% for 21 days of therapy period in the immunotherapy study.

Conclusion: Immunoglobulin Y can be a prophylactic and therapeutic agent against streptococcal infections caused E. faecalis in red tilapia with an optimum dosage of 2.43 mg/g pellet.

Keywords: Enterococcus faecalis, immunoglobulin Y, immunoprophylaxis, immunotherapy, red tilapia, streptococcal infection.

Antibiotic resistance of Escherichia coli from the milk of Ettawa crossbred dairy goats in Blitar Regency, East Java, Indonesia

Research (Published online: 27-01-2023)

21. Antibiotic resistance of Escherichia coli from the milk of Ettawa crossbred dairy goats in Blitar Regency, East Java, Indonesia

Tweedekharis Marlin Agatha, Prima Ayu Wibawati, Reza Ikhza Izulhaq, Bodhi Agustono, Ragil Angga Prastiya, Dhandy Koesoemo Wardhana, Abzal Abdramanov, Widya Paramita Lokapirnasari, and Mirni Lamid

Veterinary World, 16(1): 168-174

ABSTRACT

Background and Aim: Antimicrobial resistance, especially antibiotic resistance, is one of the most severe public health challenges. Antibiotic resistance occurs when bacteria avoid and fight the mechanism of action of antibiotic drugs. This study aimed to determine the resistance of Escherichia coli from the milk of Ettawa crossbreed dairy goat at Blitar Regency, East Java, Indonesia, with the antibiotics streptomycin, sulfonamides, and trimethoprim.

Materials and Methods: A total of 34 milk samples of Ettawa crossbreed dairy goats were used in this study. The initial stages of this research included tests of the physical properties, isolation, and identification of E. coli. Then, the E. coli isolates were tested for antibiotic resistance using the Kirby–Bauer method.

Results: The results showed that all samples were positive for E. coli. The physical properties of milk, namely, color, odor, flavor, and consistency, were normal. The results of the alcohol test showed normal acidity, and the specific gravity of goat milk met the criteria, with an average specific gravity of 1.0295 g/mL. The results of the antibiotic resistance test showed that 4 (12%) samples were resistant to streptomycin, 5 (15%) to sulfonamide, and 3% to trimethoprim.

Conclusion: The prevalence of E. coli from Ettawa crossbreed dairy goats in Blitar Regency, East Java, Indonesia, was 100%. Furthermore, this E. coli isolate exhibited resistance to antibiotics streptomycin, sulfonamides, and trimethoprim. The use of antibiotics in the dairy goat industry in Indonesia should be controlled to prevent the spread of resistant E. coli from animals to humans through the food chain and prevent the emergence of multidrug-resistant E. coli.

Keywords: antibiotic drugs, Escherichia coli, Indonesia, public health, specific gravity.