Comparison of the oxidative respiratory burst and mitogen-induced leukocyte responses of camels, goats, sheep, and cows

Research (Published online: 07-06-2022)

1. Comparison of the oxidative respiratory burst and mitogen-induced leukocyte responses of camels, goats, sheep, and cows

Abeer Al-Hamrashdi, Khalid Al-Habsi, Elshafie I. Elshafie and Eugene H. Johnson

Veterinary World, 15(6): 1398-1407

ABSTRACT

Background and Aim: The reports from the Ministry of Agriculture and Fisheries suggest that camels suffer less compared to goats, sheep, and cows from a number of common infectious diseases in Oman. However, there is no immunological evidence to substantiate this claim. This present study is, therefore, an attempt to study the immunological responses of camels, goats, sheep, and cows by comparing their oxidative respiratory burst of peripheral blood leukocytes (PBLs) as a marker of innate immunity occurring during phagocytosis and the mitogenic responses of their peripheral blood mononuclear leukocytes (PBMLs) as a marker of their adaptive immune response.

Materials and Methods: Ten female adult animals (n = 10) were selected from each species (goats, sheep, and cows). The goats, sheep, and cows were maintained at the Agricultural Experiment Station, while camels were kept at the Royal Camel Corps (RCC). Blood samples were collected from the jugular vein in 7 mL of heparin and ethylenediaminetetraacetic acid vacutainer tubes. The oxidative respiratory burst of PBLs was measured using a chemiluminescence (CL) assay. Reactants consisted of 75 μL of whole blood diluted (1:50), 75 μL of luminol/isoluminol, and 75 μL of zymosan opsonized with non-heat inactivated serum/heat-inactivated serum or non-opsonized zymosan. CL responses were measured as relative light units and expressed as the mean count per minute and peak CL values. The mitogenic response of PBMLs to concanavalin A (Con-A), phytohemagglutinin (PHA), and pokeweed mitogen (PWM) was tested using a WST-8 assay and read spectrophotometrically at 450 nm.

Results: The present findings showed that camel PBLs generate significantly higher CL responses, both intracellularly as well as extracellularly, with zymosan opsonized with autologous serum. Camel PBLs demonstrated a significantly higher (p = 0.001) response when stimulated with zymosan opsonized with heat-inactivated serum compared to those of goat, sheep, and cow lymphocytes from camels exhibited significantly higher (p = 0.001) stimulation indices (SI) with Con-A, PHA, and PWM.

Conclusion: The present study suggests that camels are capable of mounting both superior innate as well as adaptive immune responses and provide immunological evidence supporting the belief of some authors, who have proposed that camels are less susceptible to a number of common infectious diseases than other domesticated ruminants.

Keywords: adaptive immune system, chemiluminescence, concanavalin A, innate immune system, phytohemagglutinin, pokeweed mitogen.

Comparative diagnosis of bovine tuberculosis using single intradermal cervical tuberculin technique, conventional methods, enzyme-linked immunosorbent assay, and the gamma-interferon assay

Research (Published online: 31-05-2022)

30. Comparative diagnosis of bovine tuberculosis using single intradermal cervical tuberculin technique, conventional methods, enzyme-linked immunosorbent assay, and the gamma-interferon assay

Sahar Hussein Abdalla Hekal, Amany N. Dapgh, Mai Badr-Eldien Abd-Elhafeez, Hassan Mohamed Sobhy and Fatma Ahmed Khalifa

Veterinary World, 15(5): 1391-1397

ABSTRACT

Background and Aim: Bovine tuberculosis (TB) is a zoonotic disease that causes huge economic losses. This study aimed to compare the result obtained from the single intradermal test, conventional methods (culture and microscopy), gamma-interferon (IFN-γ) assay, and indirect enzyme-linked immunosorbent assay (ELISA) to diagnose bovine TB.

Materials and Methods: This study evaluated 2913 animals from milk farms in Cairo, El-Sharkia, and El-Qalyubia Governorates by single intradermal cervical tuberculin technique (SICTT), ELISA, and IFN-γ assay.

Results: Of the 2913 dairy cows surveyed, 3.7% yielded positive results. Culture prepared samples on Lowenstein-Jensen and Middlebrook 7H10 agar media yielded 52 (1.85%) isolates of Mycobacterium spp. from 2805 milk samples that yielded negative tuberculin reactions and 56 (51.85%) isolates of Mycobacterium spp. were recovered from 108 lymph node samples from positive cases. ELISA analysis of the sera of 108 positive SICTT reactors revealed that 94 (87.03%) and 97 (89.81%) animals were positive for bovine purified protein derivative (PPD-B) antigen and commercial polypeptide antigen, respectively. IFN-γ assays were performed on whole blood samples collected from positive SICTT reactors and showed that 103 (95.37%) animals were positive.

Conclusion: M. tuberculosis complex may be isolated from raw milk and not all infected animals shed mycobacterial bacilli in their milk. The use of polypeptide antigen in ELISA provides better diagnostic efficacy than PPD-B antigen. The IFN-γ assay is more sensitive than both SICTT and ELISA. It should be used in parallel with SICTT to allow the detection of more positive animals before they become a source of infection to other animals and humans.

Keywords: bovine tuberculosis, enzyme-linked immunosorbent assay, Gamma-interferon assay, Mycobacterium tuberculosis, single intradermal cervical tuberculin skin test.

Preparation and the assessed efficacy of oral baits for the vaccination of free-roaming dogs against rabies

Research (Published online: 31-05-2022)

29. Preparation and the assessed efficacy of oral baits for the vaccination of free-roaming dogs against rabies

Naglaa I. Aly, Yasser F. Elnaker, Zeinab T. S. Salama, Mohamed S. Diab, Eman A. Saber, Sotohy A. Sotohy, Wael K. Elfeil and Mohamed H. Khodeir

Veterinary World, 15(5): 1383-1390

ABSTRACT

Background and Aim: Rabies is considered a highly fatal zoonotic disease and many deaths in humans have been associated with dog bites. This study was designed to prepare an oral anti-rabies vaccine in the form of baits to eliminate the disease in free-roaming dogs and subsequently protect humans from dog bites.

Materials and Methods: The Evelyn Rokintniki Abelseth (ERA) rabies virus strain was propagated in baby hamster kidney cell cultures and adjusted to the recommended dose for application. Four forms of oral baits were employed with the rabies vaccine, which was evaluated for safety, acceptability, and potency in different dog groups. Enzyme-Linked Immunosorbent Assay (ELISA) and the serum neutralization test (SNT) were used to determine the protective rabies antibody titer in the sera of vaccinated dogs.

Results: According to the results, a dose of 3 mL of the ERA strain, containing a viral titer of 107.6 TCID50/mL, induced a mean antibody titer of 25.6 by SNT, and the PI% was 75.7 by Block ELISA, providing a protective level of the rabies antibody in 100% of vaccinated dogs. All used baits were found to be safe, inducing no abnormal general post-vaccination signs (the signs are limited to mild fever, mild loss of appetite, and mild-to-moderate loss of energy for 24-36 h after vaccination).

Conclusion: It was found that most of the accepted and highly potent bait types consisted of a mixture of wheat flour, vegetable oil, sodium alginate, corn starch, meat meal, cellulose gum, and water. This dog meal was covered with bran and edible wax to seal the bait cavity after inserting the vaccine sachet. This bait was able to induce a protective level of rabies antibodies in 100% of vaccinated dogs after receiving one bait/dog. Hence, such a bait could be recommended for use in the protection of free-roaming dogs and the elimination of the disease.

Keywords: enzyme-linked immunosorbent assay, oral baits, potency, rabies, serum neutralization test.

The activity of Meniran (Phyllanthus niruri Linn.) extract on Salmonella pullorum infected broilers

Research (Published online: 31-05-2022)

28. The activity of Meniran (Phyllanthus niruri Linn.) extract on Salmonella pullorum infected broilers

Sri Hidanah, Emy Koestanti Sabdoningrum, Kadek Rachmawati, Soeharsono Soeharsono, Gede Govinda Ananta Trika, Masy' Ariel Huda and Tsania Putri Widiati

Veterinary World, 15(5): 1373-1382

ABSTRACT

Background and Aim: Pullorum is an acute and chronic disease caused by Salmonella pullorum, often infecting chicken farms. Pullorum disease treatment using antibiotics that do not follow the control dose can cause bacteria to become antibiotic-resistant. Meniran contributes to inhibiting and antagonizing bacteria and can increase the efficiency of chicken feed because of its bioactive compounds, including alkaloids, flavonoids, tannins, and saponins. This study aimed to determine the activity of Meniran extract (Phyllanthus niruri Linn.) in broilers infected with S. pullorum.

Materials and Methods: In vitro study that was conducted includes phytochemical test, diffusion, and dilution methods using Meniran extract at 5%, 10%, 20%, and 40% concentrations and tylosin at 2% concentration. The data of the dilution method (minimum inhibitory concentration [MIC] and minimum bactericidal concentration [MBC]) were processed using probit analysis to determine LC50. In vivo study was conducted by randomly dividing 20 broilers into five treatment groups, four per group. The chickens (except in group P0–) were infected with S. pullorum aged 14 days. Then, the treatment was conducted according to the divided groups when the chickens were aged 21-34 days. The said treatments are P0– (uninfected S. pullorum and unadministered with Meniran extract), P0+ (infected with S. pullorum and unadministered with Meniran extract), and P1, P2, and P3 (infected with S. pullorum and administered with Meniran extract with 5%, 10%, and 20% concentrations, respectively). Data from the phytochemical test were analyzed as descriptive. The data from the diffusion method were analyzed using one-way analysis of variance (ANOVA) and Duncan's test. Then, the results of broilers' performance were analyzed using ANOVA and Duncan's test.

Results: The phytochemical test showed positive for alkaloid, tannin, saponin, flavonoid, and steroid/triterpenoid. The diffusion method formed the largest zone at 40% concentration with 15.6 mm, while 20%, 10%, and 5% had average of 13.15 mm, 8.38 mm, and 5.8 mm, respectively. The dilution method (MIC and MBC) exhibited the antibacterial ability of Meniran extract against S. pullorum at 20% dose and LC50 14.118% concentration. The Meniran extract administration in broilers exhibited improved performance of chickens infected with S. pullorum, with the administration of 20% dose of Meniran extract showing the best result.

Conclusion: About 20% concentration Meniran extract can serve as an antibacterial agent and showed the best results in broilers infected with S. pullorum.

Keywords: broiler chicken performance, food security, Phyllanthus niruri Linn., Salmonella pullorum.

Effect of L-arginine and L-Lysine HCl ratio on growth performance and ileum morphology of native chickens aged 2-14 weeks

Research (Published online: 27-05-2022)

27. Effect of L-arginine and L-Lysine HCl ratio on growth performance and ileum morphology of native chickens aged 2-14 weeks

Charles Venirius Lisnahan, Oktovianus R. Nahak, Welsiliana Welsiliana and Lukas Pardosi

Veterinary World, 15(5): 1365-1372

ABSTRACT

Background and Aim: Micronutrients such as essential amino acids in chicken feed must be balanced to promote optimal development. The balance of the amino acids arginine and lysine in chicken feed is particularly important. This study aimed to examine the effect of the ratio of L-arginine to L-Lysine HCl on growth performance and ileum morphology of native chickens aged 2-14 weeks-old.

Materials and Methods: One hundred and eighty 2-week-old native chickens which initial weight 78.10±4.97 g were classified into six treatments and five repetitions using a completely randomized design. Treatments were based on the ratio of arginine to lysine in the feed: T1 (0.50% L-arginine: 0.85% L-lysine HCl); T2 (0.75% L-arginine: 0.85% L-lysine HCl); T3 (1.00% L-arginine: 0.85% L-lysine HCl); T4 (0.50% L-arginine: 1.00% L-lysine HCl); T5 (0.75% L-arginine: 1.00% L-lysine HCl); and T6 (1.00% L-arginine: 1.00% L-lysine HCl).

Results: Groups T3 and T6 had the highest feed consumption (42.06±0.29 and 42.78±0.72 g/bird/day, respectively), while Group T6 had the highest body weight and body weight gain rate (1505.60±103.20 kg/bird and 16.99±1.24 g/bird/day, respectively). Groups T3 and T6 also had the highest carcass weight (916.16±46.99 and 947.18±62.32 g/bird, respectively). The best feed conversion was seen for Groups T3, T5, and T6 (2.55±0.14, 2.50±0.20, and 2.53±0.19, respectively). For ileum morphometry, the highest villus height occurred in Groups T2, T3, T5, and T6 (962.80±23.31, 982.80±10.03, 972.80±18.99, and 989.80±10.69 μm, respectively); and Group T6 had the highest crypt depth and villus width (340.80±11.52 and 302.00±4.00 μm, respectively). Statistical analysis indicated significant differences among the treatment groups for all variables examined (p<0.05).

Conclusion: The highest ratio of arginine-lysine was associated with the largest increase in native chicken feed consumption, body weight gain, feed conversion, and carcass weight, as well as villus height and width, and crypt depth in the ileum. Overall, an arginine-lysine ratio of 0.8-1.20 promoted optimal growth of native chickens aged 2-14 weeks. In the future, it is important to increase the arginine-lysine ratio with low feed protein levels in native chickens.

Keywords: growth performance, ileum morphology, L-arginine, L-lysine HCl, native chickens.

Amelioration of aflatoxin acute hepatitis rat model by bone marrow mesenchymal stem cells and their hepatogenic differentiation

Research (Published online: 27-05-2022)

26. Amelioration of aflatoxin acute hepatitis rat model by bone marrow mesenchymal stem cells and their hepatogenic differentiation

Faten A. M. Abo-Aziza, Abdel Kader A. Zaki, Rana M. Adel and Ahmed Fotouh

Veterinary World, 15(5): 1347-1364

ABSTRACT

Background and Aim: Bone marrow-derived mesenchymal stem cells (BM-MSCs) transplantation and their hepatogenic differentiated cells (HDCs) can be applied for liver injury repair by tissue grafting. Regenerative potentiality in liver cirrhosis models was widely investigated; however, immunomodulation and anti-inflammation in acute hepatitis remain unexplored. This study aimed to explore the immunomodulatory and evaluate twice intravenous (IV) or intrahepatic (IH) administration of either BM-MSCs or middle-stage HDCs on aflatoxin (AF) acute hepatitis rat model.

Materials and Methods: BM-MSCs viability, phenotypes, and proliferation were evaluated. Hepatogenic differentiation, albumin, and a-fetoprotein gene expression were assessed. AF acute hepatitis was induced in rats using AFB1 supplementation. The transplantation of BM-MSCs or their HDCs was done either by IV or IH route. Hepatic ultrasound was performed after 3-weeks of therapy. Cytokines profile (tumor necrosis factor-α [TNF-α], interleukin [IL]-4, and IL-10) was assessed. Hepatic bio-indices, serum, and hepatic antioxidant activity were evaluated, besides examining liver histological sections.

Results: Acute AFB1 showed a significant increase in TNF-α (p<0.01), liver enzyme activities (p<0.05), as well as decrease in IL-4, IL-10, and antioxidant enzyme activities (p<0.05). Cytokines profile was ameliorated in groups treated with IV and IH BM-MCs, showed a negative correlation between IL-4 and TNF-α (p<0.05), and a positive correlation between IL-10 upregulation and TNF-α (p<0.01). In IV HDCs treated group, positive correlations between IL-4 and IL-10 downregulation and TNF-α were observed. However, in IH HDCs group, a significant positive correlation between IL-4 and IL-10 upregulation and TNF-α, were recorded (p<0.05). In addition, IV BM-MSCs and IH HDCs treatments significantly increased antioxidant enzymes activity (p<0.05). IV and IH BM-MSCs significantly ameliorated liver transaminase levels, whereas IH HDCs significantly ameliorated alanine aminotransferase activity and nitric oxide concentration (p<0.05).

Conclusion: The administration routes of BM-MSCs did not demonstrate any significant difference; however, the IH route of HDCs showed significant amelioration from the IV route. On the other hand, it showed noticeable anti-inflammatory and immunomodulatory improvements in aflatoxicosis rats. Therefore, it can be concluded that acute hepatitis can be treated by a noninvasive IV route without the expense of hepatogenic differentiation. Further research using clinical trials that address several problems regarding engraftment and potentiation are needed to determine the optimal manipulation strategy as well as to achieve better long-term effects.

Keywords: acute hepatitis, bone marrow mesenchymal stem cell, cytokines, hepatogenic differentiation, rat model.

First survey on seroprevalence of Japanese encephalitis in long-tailed macaques (Macaca fascicularis) in Bali, Indonesia

Research (Published online: 27-05-2022)

25. First survey on seroprevalence of Japanese encephalitis in long-tailed macaques (Macaca fascicularis) in Bali, Indonesia

I Gusti Agung Arta Putra, Anak Agung Ayu Mirah Adi, I Nyoman Mantik Astawa, I Made Kardena, I Nengah Wandia, I Gede Soma, Fany Brotcorne and Agustin Fuentes

Veterinary World, 15(5): 1341-1346

ABSTRACT

Background and Aim: Japanese encephalitis (JE) is a zoonotic infectious inflammatory brain disease caused by the JE virus (JEV). Considerable research into the seroprevalence of JE in domestic animals has been conducted, but there have been no reports of its occurrence in wild animals, including long-tailed macaques (Macaca fascicularis). This study aimed to estimate the seroprevalence of JEV infection and its determinants in long-tailed macaques in Bali and the prevalence of mosquito vectors.

Materials and Methods: Blood samples (3 mL) were collected from a population of M. fascicularis (92 heads) inhabiting a small forest with irrigated rice field nearby (wetland area) in Ubud, Gianyar, and from two populations in dryland areas with no wet rice field (Uluwatu, Badung, and Nusa Penida, Bali Province, Indonesia). The collected sera were tested for antibodies against JEV using a commercially available enzyme-linked immunosorbent assay kit (qualitative monkey JE Immunoglobulin G antibody kit). The seropositivity of the antibodies was then compared based on different variables, namely, habitat type, age, and sex.

Results: The seroprevalence of the JEV antibodies in all the samples tested was found to be 41.3%. The seropositivity of the monkey serum samples collected from the wetland area was 46.4%, which was higher than the seropositivity of the sera samples collected from the dried field areas (1.25%). Monkey sera collected from the wetland areas were 6.1 times (odds ratio [OR]: 6.1; 95% confidence interval [CI]: 0.71-51.5, p>0.05) more likely to be seropositive compared to the monkey sera collected from the dried field areas. Meanwhile, female monkeys were 1.79 times (OR: 1.79; 95% CI: 0.76-4.21; p>0.05) more likely to be seropositive to JEV than males. Similarly, juvenile monkeys were 2.38 times (OR: 2.38; 95% CI: 0.98-5.79); p>0.05) more likely to be seropositive against the JEV than adult monkeys. However, none of these differences achieved statistical significance. Regarding the JEV mosquito vector collection, more Culex mosquitoes were found in the samples from the wetland areas than from the dried field areas.

Conclusion: The study confirms the existence of JEV infection in long-tailed macaques in Bali. There were patterned seropositivity differences based on habitat, age, and sex of the monkeys, but these were not significant. The possibility of monkeys as a JEV reservoir and the presence of the mosquitoes as the JEV vector are suggested but require more study to confirm.

Keywords: enzyme-linked immunosorbent assay, Japanese encephalitis virus, Macaca fascicularis, seroprevalence.