Angiotensin-converting enzyme inhibitor activity of peptides derived from Kacang goat skin collagen through thermolysin hydrolysis

Research (Published online: 21-01-2021)

21. Angiotensin-converting enzyme inhibitor activity of peptides derived from Kacang goat skin collagen through thermolysin hydrolysis

Arby'in Pratiwi, Thoyib R. Hakim, Mohammad Z. Abidin, Nanung A. Fitriyanto, Jamhari Jamhari, Rusman Rusman and Yuny Erwanto

Veterinary World, 14(1): 161-167

ABSTRACT

Background and Aim: Angiotensin-converting enzyme (ACE) is one of the inhibitory enzymes isolated from animals for the treatment of hypertension. ACE inhibitor (ACE-I) peptides can be obtained by hydrolyzing proteins from various animal tissues, including muscle and connective tissues. However, the study on ACE-I activity from collagen of Kacang goat skin has not been conducted. This study explores the potency of collagen from Kacang goat skin as a source of an antihypertensive agent through ACE inhibition. Thermolysin will hydrolyze collagen and produce the peptide classified antihypertensive bioactive peptides. This study aimed to determine the potential of thermolysin to hydrolyze collagen of Kacang goat skin for ACE-I peptide production and to identify the production of ACE-I peptides.

Materials and Methods: Collagen from Kacang goat skin was hydrolyzed with thermolysin and incubated at 37°C for 1 h. Molecular weight (MW) evaluation was performed by SDS PAGE; fractionation peptides at <5 kDa, 3-5 kDa, and <3 kDa were performed by ultrafiltration and ACE-I activity determined by IC50 measurement.

Results: Collagen was hydrolyzed by thermolysin, resulting in protein with MW of 117.50-14.60 kDa. The protein content of fractionation at >5 kDa was 3.93±0.72 mg/mL, content of 3-5 kDa was 3.81±0.68 mg/mL, and that of <3 kDa was 2.33±0.38 mg/mL. Fractionation was performed 3 times and one of the results was selected for the ACE-I test. The selected fraction was tested by IC50 measurement with three repetitions and it showed an average enzyme activity at 0.83 μg/mL or 82.94 mg/mL.

Conclusion: Thermolysin hydrolysis of collagen from Kacang goat skin showed the potential to produce bioactive peptides, such as ACE-I.

Keywords: angiotensin-I-converting enzyme inhibitor, bioactive peptides, collagen, hydrolysis, thermolysin.

Cysticercus bovis in cattle slaughtered in North Egypt: Overestimation by the visual inspection method

Research (Published online: 20-01-2021)

20. Cysticercus bovis in cattle slaughtered in North Egypt: Overestimation by the visual inspection method

Mona Hassan El-Sayad, Hoda Farag, Hend El-Taweel, Reda Fadly, Nahla Salama, Asmaa Abd Elhameed Ahmed and Naglaa Fathi Abd El-Latif

Veterinary World, 14(1): 155-160

ABSTRACT

Background and Aim: The World Health Organization and the Food and Agriculture Organization list Taenia saginata, a foodborne cestode, as the most widely distributed human tapeworm worldwide. The larval stage of T. saginata, Cysticercus bovis, causes cysticercosis in bovines and infects humans who eat raw or undercooked beef. The existing detection methods of C. bovis in cattle depend on the visual inspection of meat. This study aimed to confirm the identification of C. bovis through visual inspection at the slaughterhouses in North Egypt with a molecular diagnosis.

Materials and Methods: A total of 687 locally bred cattle (Baladi), including 428 cows and 259 buffaloes, slaughtered in four slaughterhouses in North Egypt from April 2018 to February 2019 were inspected for C. bovis using the traditional meat inspection method. Positive samples were verified through polymerase chain reaction (PCR) amplification and HDP2 gene sequencing.

Results: Through visual inspection, C. bovis was detected in 4.2% and 12.4% of the slaughtered cows and buffaloes, respectively. Molecular analysis confirmed that 1.9% of the animals, all of which were cows, had C. bovis infection. DNA sequencing verified the identity of the PCR-amplified product.

Conclusion: The rate of C. bovis infection in slaughterhouses detected through meat inspection is overestimated compared with that through PCR. Although meat inspection can be used as a primary screening tool for C. bovis, a more specific molecular method is required to achieve an accurate diagnosis.

Keywords: cattle, Cysticercus bovis polymerase chain reaction analysis, Taenia saginata, zoonotic.

Validation of real-time polymerase chain reaction versus conventional polymerase chain reaction for diagnosis of brucellosis in cattle sera

Research (Published online: 19-01-2021)

19. Validation of real-time polymerase chain reaction versus conventional polymerase chain reaction for diagnosis of brucellosis in cattle sera

Nour H. Abdel-Hamid, Eman I. M. Beleta, Mohamed A. Kelany, Rania I. Ismail, Nadia A. Shalaby and Manal H. M. Khafagi

Veterinary World, 14(1): 144-154

ABSTRACT

Background and Aim: Different polymerase chain reaction (PCR) techniques have and are still being used for the direct detection of Brucella DNA in serum samples of different animal species and humans without being validated or properly validated, resulting in discrepancies. Thus, this study aimed to evaluate the diagnostic performance of the TaqMan Real- Time-PCR (RT-PCR) targeting the bcsp31 gene versus conventional PCR for the accurate diagnosis of brucellosis at the genus level in cattle sera.

Materials and Methods: One hundred and eighty-four serum samples were collected from bacteriologically positive and negative cows with ages ranging from 1 to 5 years old at some infected private farms in the Nile Delta under quarantine measures as well as brucellosis free farms. These samples were classified into four groups after serological diagnosis and investigated by TaqMan RT-PCR and conventional PCR targeting the IS711 gene for Brucella DNA detection. The diagnostic performance characteristics of both PCR techniques were estimated considering the bacteriological results as a gold standard.

Results: TaqMan RT-PCR revealed superiority over conventional PCR; it was able to detect Brucella DNA in 95% (67/70) and 89% (25/28) of the cattle sera samples belonging to Group 1 (serologically and bacteriologically positive) and Group 2 (serologically negative but bacteriologically positive), respectively. On evaluating the diagnostic performance, TaqMan RT-PCR showed superior diagnostic sensitivity (93.9%), diagnostic specificity (88.4%), performance index (182.3), almost perfect kappa agreement (0.825±0.042), strong positive correlation (r=0.826), high accuracy based on the receiver operating characteristic (ROC) curve, and area under the ROC curve (0.911) at p<0.05 and CI of 95%.

Conclusion: A cattle serum sample is not the metric of choice for targeting Brucella genomic DNA by conventional PCR. The time-saving and rapid TaqMan RT-PCR method revealed a better diagnostic performance in the detection of Brucella DNA in cattle sera. Such performance offered by TaqMan RT-PCR may be considered a step toward the possibility of using such technology in the direct differentiation between Brucella-infected and -vaccinated cattle immunized by smooth vaccines from cattle sera using primers specific for such vaccines.

Keywords: bacteriological results, Brucella, conventional polymerase chain reaction, diagnostic sensitivity, diagnostic specificity, TaqMan real-time-polymerase chain reaction.

Amelioration on oxidative stress, testosterone, and cortisol levels after administration of Vitamins C and E in albino rats with chronic variable stress

Research (Published online: 18-01-2021)

18. Amelioration on oxidative stress, testosterone, and cortisol levels after administration of Vitamins C and E in albino rats with chronic variable stress

Nanik Hidayatik, Agus Purnomo, Faisal Fikri and Muhammad Thohawi Elziyad Purnama

Veterinary World, 14(1): 137-143

ABSTRACT

Background and Aim: Stress can cause physiological and biological disorders in the body. On the other hand, antioxidants from vitamins and minerals are effective for stress treatment. Therefore, this study aimed to evaluate the effect of the administration of Vitamins C and E on serum superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT), glutathione peroxidase (GPx), testosterone, and cortisol activity in albino rats with chronic variable stress (CVS).

Materials and Methods: Twenty albino rats were randomly assigned into four treatment groups: C was administered normal saline; T1 was administered Vitamins C and E; T2 was only induced CVS; and T3 was induced CVS followed by Vitamins C and E administration. All treatments were applied for 4 weeks, respectively. Furthermore, 5 mL of blood samples were collected intracardially. Body weight data were collected for the initial and final weights. From serum samples, SOD, GPx, and CAT were measured using the enzymol method; MDA was measured using the high-performance liquid chromatography method; and testosterone and cortisol were measured using the enzyme-linked immunosorbent assay method. All variables were analyzed statistically using analysis of variance followed by the Duncan test (p<0.05).

Results: Our findings showed that the T1 and T3 groups significantly decreased (p<0.001) compared to T2 in the following parameters: SOD, MDA, GPx, and cortisol. Meanwhile, CAT and testosterone levels in the T1 and T3 groups were significantly increased (p<0.001) compared to the T2 group. In addition, the weight gain in T1 and T3 groups was significantly increased (p<0.001) compared to T2 group.

Conclusion: It can be concluded that the administration of Vitamins C and E had a significant effect to alleviate SOD, MDA, GPx, and cortisol and to improve the testosterone level in albino rats with CVS.

Keywords: chronic variable stress, oxidative stress parameters, Vitamin C, Vitamin E.

Obstructive urolithiasis in buffalo calves (Bubalus bubalis): Serum changes of Vitamins A and D and efficacy of surgical management using tube cystostomy

Research (Published online: 18-01-2021)

17. Obstructive urolithiasis in buffalo calves (Bubalus bubalis): Serum changes of Vitamins A and D and efficacy of surgical management using tube cystostomy

Abdelmonem Abdallah, Shimaa Ezzeldein, Eslam Eisa, Mustafa Abd El Raouf and Yasmin Bayoumi

Veterinary World, 14(1): 129-136

ABSTRACT

Background and Aim: Obstructive urolithiasis is one of the major health problems in livestock animals, mainly in young calves. The present study was designed first to investigate the changes in the serum levels of Vitamins A and D in buffalo calves (Bubalus bubalis) with obstructive urolithiasis and second to investigate the efficacy of tube cystostomy technique in management of such condition.

Materials and Methods: One hundred and forty-nine buffalo calves of variable ages ranged from 3 to 7 months with a history of retained urine were examined clinically and ultrasonographically. Then, they were subjected to surgical treatment using the tube cystostomy technique. The serum levels of Vitamins A and D were investigated in retained urine calves in addition to 10 clinically healthy calves of the same age used as a control group.

Results: Based on clinical and ultrasonographic findings, the calves were diagnosed as obstructive urolithiasis with intact bladder (n=64 calves) or with bladder rupture (n=85 calves) with the peak incidence in winter months. Tube cystostomy was an efficient and quick surgical technique for the management of such condition and 95.3% of calves returned their normal urination within 7–14 days after surgery. Significant hypovitaminoses A and D were found between retained urine calves and control ones (p= 0.01 and 0.002, respectively).

Conclusion: Hypovitaminoses A and D suggested predisposing obstructive urolithiasis in buffalo calves, but further clinical studies are recommended for more confirmation. Surgical treatment using tube cystostomy technique is recommended for the management of obstructive urolithiasis in buffalo calves.

Keywords: buffalo calves, hypovitaminosis, tube cystostomy, ultrasound, urolithiasis.

Novel in vitro and in vivo anti-Helicobacter pylori effects of pomegranate peel ethanol extract

Research (Published online: 16-01-2021)

16. Novel in vitro and in vivo anti-Helicobacter pylori effects of pomegranate peel ethanol extract

Amal Mayyas, Mohammad Abu-Sini, Rula Amr, Rand T. Akasheh, Waleed Zalloum, Ayman Khdair, Islam Hamad, Talal Aburjai, Rula M. Darwish and Luay Abu-Qatouseh

Veterinary World, 14(1): 120-128

ABSTRACT

Background and Aim: Interest in plants with antimicrobial properties has been revived due to emerging problems associated with using antibiotics to eradicate Helicobacter pylori. Accordingly, this study aims to assess the antibacterial effects of Punica granatum and the possible synergistic effect of its extract along with metronidazole against H. pylori.

Materials and Methods: Pomegranate peel ethanol extracts (PPEE) was tested against a control strain of H. pylori (NCTC 11916) in vitro and in vivo in female Wistar rats. Moreover, the synergistic effect of PPEE in combination with metronidazole was tested in vitro.

Results: The PPEE exhibited a remarkable activity against H. pylori with a minimum inhibitory concentration (MIC) of 0.156 mg/mL. Furthermore, the extract exhibited a pronounced urease inhibitory activity (IC50 ∼6 mg/mL) against the tested strain. A synergistic effect between PPEE and metronidazole was also observed (fractional inhibitory concentrations <0.5). Oral treatment of rats with PPEE for 8 days produced a significant reduction in H. pylori gastritis and a significant decrease in both lymphocytic and positive chronicity.

Conclusion: Pomegranate extract is probably safe and represents a potential alternative and complementary therapy for reducing H. pylori associated with gastric ulcers.

Keywords: Helicobacter pylori, in vitro, in vivo, pomegranate peel ethanol extracts, urease inhibition.

Genetic and parasitological identification of Trypanosoma evansi infecting cattle in South Sulawesi, Indonesia

Research (Published online: 15-01-2021)

15. Genetic and parasitological identification of Trypanosoma evansi infecting cattle in South Sulawesi, Indonesia

Agus Setiawan, Wisnu Nurcahyo, Dwi Priyowidodo, Rina Tri Budiati and Desy Sylvia Ratna Susanti

Veterinary World, 14(1): 113-119

ABSTRACT

Background and Aim: Sulawesi is an Indonesian island located within the Wallacea region that contains a distinctive mix of Asian and Australasian species. This distinctiveness extends to parasites, including Trypanosoma evansi, the cause of surra. Surra has non-specific clinical signs such as anemia, anorexia, weight loss, drop in milk production, and reproductive disorders which cause economic losses. Due to the trade of livestock, surra has spread in Indonesia from one island to another. The aim of this study was to investigate the trypanosomes infecting cattle in South Sulawesi, using internal transcribed spacer (ITS2) ribosomal DNA (rDNA) sequencing.

Materials and Methods: A total of 100 whole blood samples were collected from cattle in Makassar, South Sulawesi Province, Indonesia. All samples were tested using conventional parasitological methods (CPT), namely, thin blood smear, buffy coat smears, and polymerase chain reaction (PCR) testing. Positive PCR results were sequenced and phylogenetically analyzed.

Results: Only one of the 100 samples was found to be positive with microscopic observation; however, PCR analysis revealed that 3% (3/100) of samples were positive. Sequencing identified the positive samples as T. evansi, China isolate (KU552344), with a homology of 99%. Two out of three sequences showed variations in ITS2 region.

Conclusion: Based on CPT and molecular analysis, T. evansi isolates from infected cattle in South Sulawesi demonstrate genetic diversity of ITS2 sequences.

Keywords: cattle, internal transcribed spacer-2, Sulawesi, Surra, Trypanosoma evansi.