Tuesday, 31 May 2022

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 LC50In 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.



Friday, 27 May 2022

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.



Thursday, 26 May 2022

Investigation of proteomic profiles in canine lymphoma using tandem mass tag-based quantitative proteomics approach

Research (Published online: 26-05-2022)
24. Investigation of proteomic profiles in canine lymphoma using tandem mass tag-based quantitative proteomics approach
Piyanoot Fonghem, Trairak Pisitkun, Kasem Rattanapinyopituk, Sirintra Sirivisoot and Anudep Rungsipipat
Veterinary World, 15(5): 1333-1340

ABSTRACT

Background and Aim: Specific tumor biomarkers are useful for the early diagnosis of cancer or can predict the recurrence of neoplastic disease in humans and animals. Lymphoma in dogs could be classified into B-, T-, and NK-cell origins. T-cell lymphoma has the worst prognosis with a shorter survival time and disease-free interval. This study aimed to identify the differential serum protein expressions of canine B- and T-cell lymphomas compared with healthy dogs using a tandem mass tag (TMT)-based quantitative proteomics.

Materials and Methods: Serum samples were collected from 20 untreated canine lymphomas (14 B-cells and 6 T-cells) and four healthy control dogs. Sera peptides from each sample were processed for TMT 10-plex tagging and analyzed using liquid chromatography-mass spectrometry (MS). Differential proteome profiling was then compared between lymphoma and control.

Results: We discovered 20 elevated and 14 decreased serum proteins in the lymphoma group relative to the healthy group. Six candidate increased proteins in canine lymphomas were beta-actin cytoplasmic 1 (ACTB, p=0.04), haptoglobin (p=0.002), beta-2 microglobulin (β2M, p=0.007), beta-2 glycoprotein 1 (APOH, p=0.03), metalloproteinase inhibitor 1 (TIMP-1, p=0.03), and CD44 antigen (p=0.02). When compared between B- and T-cell lymphomas, B-cell phenotypes had upregulated immunoglobulin (Ig) heavy chain V region GOM (p=0.02), clusterin (p=0.01), apolipoprotein C1 (APOC1, p=0.05), and plasminogen (p=0.02).

Conclusion: These findings were investigated quantitative serum proteomes between B- and T-cell lymphomas using TMT-based MS. ACTB, β2M, APOH, TIMP-1, CD44 antigen, Ig heavy chain V region GOM, and APOC1 are novel candidate proteins and might serve as a lymphoma biomarker in dogs. However, evaluation with an increased sample size is needed to confirm their diagnostic and prognostic ability.

Keywords: B-cell lymphoma, dog, tandem mass tag proteomics, T-cell lymphoma.





Success in animal skin fashion in African countries or the boom of the wet market

Review (Published online: 26-05-2022)
23. Success in animal skin fashion in African countries or the boom of the wet market
Andile Ephraim Mkhonza, Keitiretse Molefe and Odirile Thato Lebogang Ramafoko
Veterinary World, 15(5): 1328-1332

ABSTRACT

The world and the way things are done have changed, from selling clothing in brick-and-mortar stores to online shopping through social media platforms. Population growth has significantly contributed to an increased clothing demand, which, in turn, has increased the demand for animal skin. Traditional markets, also known as wet markets, are considered as major zoonotic disease reservoirs due to human and animal contact. Some groups and individuals continue to believe in traditional medicine and clothing that is made from animal skin, and such beliefs are more accessible with the presence of wet markets. Hence, animal poaching and trafficking have increased to meet the high demands, primarily in the Western world. Poverty is a well-known motivation to commit a crime. Conservationists should not only look at the animal regulation site to propose a solution to animal poaching and trafficking but should also consider communal poverty. Thus, this review aimed to highlight the role of wet market and animal skin fashion on animal welfare and human health.

Keywords: animal welfare, clothing, disease reservoirs, poverty.



First study on microscopic and molecular evidences of two bovine hemoplasma species in cattle herds in Al-Qadisiyah Province, Iraq

Research (Published online: 26-05-2022)
22. First study on microscopic and molecular evidences of two bovine hemoplasma species in cattle herds in Al-Qadisiyah Province, Iraq
Yahia Ismail Khudhair, Zeena Fouad Saleh and Hayder N. Ayyez
Veterinary World, 15(5): 1323-1327

ABSTRACT

Background and Aim: Hemotropic Mycoplasmas are small epierythrocytic bacteria that cause infectious anemia in several livestock species and in humans. Several reports have been made on hemoplasma infections in the south and north of Iraq, but there have been no studies in the middle Euphrates of Iraq. This study aimed to evaluate the presence of hemoplasma species in cattle in Al-Qadisiyah Province, Iraq.

Materials and Methods: Two hundred blood samples were collected from cattle with pale mucous membrane from regions with heavy tick endemicity. The samples were analyzed for the presence of Rickettsia pathogens using thin blood smears and the Diff-Quik stains. All the samples were also examined using polymerase chain reaction (PCR) to amplify the 16S ribosomal ribonucleic acid (rRNA) gene to confirm the presence of the smear-identified microorganisms. Ten PCR positive samples were subjected to 16S rRNA partial gene sequencing to identify the species.

Results: The findings uncovered positivity in 68 (34%) blood smears. PCR revealed positive confirmation in 18 (9%) of the 200 blood samples. Mycoplasma wenyonii and Candidatus mycoplasma hemobos were identified from 10 PCR positive samples. The nucleotide sequences of the isolates were closely related to isolates from cattle, buffalo, and dogs in Vietnam, Cuba, India, and Germany.

Conclusion: Bovine hemoplasma infections are prevalent in cattle in the Al-Qadisiyah Province in Iraq. Our results may have significance for the development of control programs.

Keywords: hemoplasma spp., microscopic, molecular diagnosis, Mycoplasma wenyonii, Iraq.



Wednesday, 25 May 2022

Effects of environmental condition, size, coat type, and body condition score on rectal temperature prediction in dogs using infrared auricular and surface temperature

Research (Published online: 25-05-2022)
21. Effects of environmental condition, size, coat type, and body condition score on rectal temperature prediction in dogs using infrared auricular and surface temperature
Yanisa Lukkanawaraporn, Nutnicha Tiangtas, Vorapan Chaikornkij, Patthamon Nawapakpilai, Sathita Areerat, Pipatpong Chundang, Chalermpol Lekcharoensuk and Attawit Kovitvadhi
Veterinary World, 15(5): 1314-1322

ABSTRACT

Background and Aim: Generally, rectal body temperature (BTrectum) is used to prefer as core body temperature in dogs. However, this procedure is time- and labor-consuming with stress induction. Therefore, infrared auricular temperature (BTear) and surface temperature (ST) could be applied to estimate BTrectum. This study aimed to estimate BTrectum from BTear or ST in various areas and determined the factors that influenced the accuracy of prediction equations.

Materials and Methods: Under controlled temperature (n=197) and ambient temperature (n=183), the parameters BTrectum, BTear, and ST at internal pinna, auricular canal, lateral aspect of shoulder, hip, axillary area, inguinal area, footpad, and anal area (STrectum) were measured. In addition, temperature and humidity levels of the surrounding environment were recorded. The correlation between each measurement technique was calculated. The BTrectum prediction equation was created using all measured data and several influencing factors (environmental condition, size, coat type, and body condition score [BCS]).

Results: The highest correlation with BTrectum was observed for BTear (r=0.61, p<0.01), which was similar to STrectum (r=0.61, p<0.01). Based on multiple linear regression model results using BTrectum as the dependent variable, BTear or STrectum were first selected as independent variables in all estimation equations. Ambient temperatures (R2=0.397), small breed (R2=0.582), long hair (R2=0.418), and/or a BCS of 2 (R2=0.557) provided the highest coefficients of determination of the prediction equation.

Conclusion: The most appropriate predictors for estimating BTrectum were STrectum and BTear, which were impacted by the dog's signalments and the environment. To obtain satisfactory outcomes, the equation must be selected depending on the dog's signalments and the environmental conditions. However, based on the findings of this investigation, the accuracy remains low in several equations, and further studies are needed to improve the accuracy of the equation, mainly by increasing the sample size and developing a specific equation for each dog's signaling and environmental condition.

Keywords: auricular temperature, body surface temperature, dog, infrared surface temperature, rectal surface temperature.



Tuesday, 24 May 2022

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 invAstnompA, 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 invAstnompA, 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.



Monday, 23 May 2022

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 alginolyticusVibrio 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.



Sunday, 22 May 2022

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.