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Molecular identification, characterization, and structure analysis of house musk shrew (Suncus murinus) leptin
2019
Sayaka Saga | Noriyasu Sasaki | Toshiro Arai
Objective: House musk shrew (Suncus murinus), a small experimental animal with low body fat, may be a possible model for human lipodystrophy. Leptin is an adipocyte-derived hormone thought to have an important role in the pathophysiology of lipodystrophy. The objectives of this study were to clarify the structure and distribution of suncus leptin. Materials and methods: To determine the primary structure of suncus leptin, we cloned the suncus Lep cDNA using the rapid amplification of cDNA ends method. The obtained amino acid (aa) sequence was compared with other mammals and the protein structure prediction was performed. Results: The suncus Lep cDNA encodes 170 aa. The putative suncus leptin precursor has a predicted signal peptide of 21 aa, and the mature leptin comprises 149 aa. The mature leptin is 75%82% homologous to that of other species. Insertion of the three aa, VPQ, not seen in other mammals was found. This VPQ insertion is thought to be due to a nucleotide insertion of nine bases by slippage-like microindels. The predicted 3D structure of suncus leptin exhibited a typical four a-helix structure, however, the VPQ region protruded compared with human leptin. LepmRNA expression was observed only in white and brown adipose tissues. Conclusion: This study revealed the structure and distribution of suncus leptin. Because the addition of VPQ, which is not found in other mammals, was observed, suncus leptin attracts attention to its physiological action, and to the possibility of being a model of human lipodystrophy. [J Adv Vet Anim Res 2019; 6(1.000): 1-8]
Show more [+] Less [-]Protective effect of avenanthramides against cisplatin induced nephrotoxicity in rats
2019
Yasmin O. El Amir | Wisam Omar | Amani Y. Khabrani | Amani E. Jahfali | Shahd M. Alhakami | Nada M. Dobab
Objective: The aim of this study is to inspect the ameliorative effect of avenanthramides (AVA) on CP nephrotoxicity in Sprague-Dawley rats. Materials and Methods: Blood samples were collected for the determination of hematological parameters. Creatinine, blood urea nitrogen (BUN), and tumor necrosis factor-α (TNF-α) were measured in serum. Specimens from both kidneys were taken for histopathological examinations. Results: Administration of AVA resulted in significant decrease in the level of creatinine and TNF-α when compared with CP group. Histopathologically, CP-induced vacuolar degeneration and necro¬sis of the kidney tubules. Administration of AVA ameliorates the histopathological alterations induced by CP. Conclusion: AVA can be considered as a protective agent for kidneys during administration of CP. The protective effect of AVA may be related to the reduction of TNF-α which implicated in the pathogenesis of CP nephrotoxicity. [J Adv Vet Anim Res 2019; 6(4.000): 521-527]
Show more [+] Less [-]The employment of q-PCR using specific primer targeting on mitochondrial cytochrome-b gene for identification of wild boar meat in meatball samples
2019
Ganea Qorry Aina | Yuny Erwanto | Motalib Hossain | Mohd Rafie Johan | Md. Eaqub Ali | Abdul Rohman
Objective: The objective of this study was to employ real-time or quantitative polymerase chain reaction (q-PCR) using novel species specific primer (SSP) targeting on mitochondrial cytochrome-b of wild boar species (CYTBWB2-wb) gene for the identification of non-halal meat of wild boar meat (WBM) in meatball products. Materials and Methods: The novel SSP of CYTBWB2-wb was designed by our group using PRIMERQUEST and NCBI software. DNA was extracted using propanol-chloroform-isoamyl alcohol method. The designed SSP was further subjected for validation protocols using DNA isolated from fresh meat and from meatball, which include specificity test, determination of efficiency, limit of detection and repeatability, and application of developed method for analysis of commercially meatball samples Results: The results showed that CYTBWB2-wb was specific to wild boar species against other animal species with optimized annealing temperature of 59°C. The efficiency of q-PCR obtained was 91.9% which is acceptable according to the Codex Allimentarius Commission (2010). DNA, with as low as 5 pg/μl, could be detected using q-PCR with primer of CYTBWB2-wb. The developed method was also used for DNA analysis extracted from meatball samples commercially available. Conclusion: q-PCR using CYTBWB2-wb primers targeting on mitochondrial cytochrome-b gene (forward: CGG TTC CCT CTT AGG CAT TT; Reverse: GGA TGA ACA GGC AGA TGA AGA) can be fruitfully used for the analysis of WBM in commercial meatball samples. [J Adv Vet Anim Res 2019; 6(3.000): 300-307]
Show more [+] Less [-]Evaluation of wound healing potential of pomegranate (Punica granatum) whole fruit extract on skin burn wound in rats (Rattus norvegicus)
2019
Bambang Sektiari Lukiswanto | Alya Miranti | Sri Agus Sudjarwo | Hardany Primarizky | Wiwik Misaco Yuniarti
Objective: This research was conducted to study the wound healing process of whole fruit pome¬granate extract (punica granatum) standardized with 40% ellagic acid ointment for deep sec¬ond-degree burn wound of skin in the rat (Rattus norvegicus). Materials and Methods: Powder of standardized pomegranate extract (SPE) with 40% ellagic acid was processed to become ointments. Twenty-five male rats, weighed 150180 gm at 3 months of age, were randomly divided into five groups. After anesthetized, stainless circle plate with 1 cm of diameter in 85°C was contacted firmly toward right gluteal of rat skins for 5 sec in order to create deep second-degree burn wound. Control groups consist of (T0) cream base and (T1) 1% silver sulfadiazine. Treatment groups consist of (T2) 2.5% SPE, (T3) 5% SPE, and (T4) 10% SPE. Histopathological preparation used hematoxylin-eosin stained skin samples. Histological observa¬tions were performed using the optics microscope against collagen, the number of polymorpho¬nuclear cell (PMN) infiltration, the degree of angiogenesis, and re-epithelization. The results were statistically compared between groups. Results: Microscopic observation on the wound healing process on the collagen, PMN infiltration, angiogenesis, and re-epithelization showed that topical administration of 10% SPE in burns gives the best result. This is characterized by a high density of collagen with a good arrangement, which is accompanied by a complete and mature epithelium, low number of inflammatory cells, and angiogenesis. This may be caused by the compounds in the pomegranate extract, which have the antioxidant, anti-inflammatory, and anti-bacterial effects. Conclusion: This study reveals that 10% SPE accelerates the healing of deep second-degree burn wound. Thus, pomegranate standardized with 40% ellagic acid is a promising herb for the healing of burn wound of skin. [J Adv Vet Anim Res 2019; 6(2.000): 202-207]
Show more [+] Less [-]Prevalence and phylogenetic relationship among methicillin- and vancomycin-resistant Staphylococci isolated from hospitals dairy food, food handlers, and patients
2019
Mona Ahmed El-Zamkan | Asmaa Gaber Mubarak | Alsagher Omran Ali
Objective: The aim of the present work was to investigate the mutual role that may be played by the served dairy food and food handlers in the transmission of methicillin- and vancomycin-resistant Staphylococcus aureus and coagulase-negative Staphylococci to patients who were hospitalized in Qena City, Egypt. Materials and Methods: A total of 210 samples including 90 dairy food samples which offered to the patients in the hospital, 60 nasal and hand swabs from food handlers working in the hospital, and 60 nasal and diarrheal swabs from patients suffering from diarrhea were investigated for the presence of coagulase-positive S. aureus and coagulase-negative Staphylococci, then isolates were screened for methicillin and vancomycin resistance phenotypically and genotypically. 16s rRNA gene sequencing was employed to construct the neighbor-joining tree. Results: Unlike food samples, both coagulase-positive S. aureus and coagulase-negative Staphylococci occurred in human samples. Methicillin- and vancomycin-resistant coagulase-negative Staphylococci could be detected in 41.7% & 20.8%, 68% & 31.9%, and 81.3% & 55.2% of iso¬lates obtained from dairy food, food handlers, and patients samples, respectively. Whereas 81% & 64.3%, and 75.4% & 38.6% of coagulase-positive S. aureus obtained from food handlers and patients samples exhibited resistance to methicillin and vancomycin, respectively. Phenotypic resistance was confirmed molecularly through detection of mecA and vanA genes. Conclusion: A significant role can be played by food and food handlers in the transmission of methicillin- and vancomycin-resistant Staphylococci to patients, which has been proved in this study through the close phylogenetic relation between S. epidermidis isolated from food, food handlers, and patients diarrheal samples. [J Adv Vet Anim Res 2019; 6(4.000): 463-473]
Show more [+] Less [-]Antibiotic susceptibility of Staphylococci isolated from bovine mastitis in Algeria
2019
Radhwane Saidi | Nora Mimoune | Ratiba Baazizi | Mohamed Hocine Benaissa | Djamel Khelef | Rachid Kaidi
Objective: This work aimed to determine the resistance and/or the susceptibility to antibiotics of staphylococci isolated from cattle with mastitis in the North of Algeria. Materials and Methods: The disk diffusion method was carried out to reveal the antibiotic resis¬tance in accordance to the National Committee for Clinical Laboratory Standards guidelines in the Mueller-Hinton agar. Results: Coagulase-negative Staphylococci (CNS) isolates showed more resistance to Cefoxitin, Amoxicillin + Clavulanic Acid, Vancomycin, Trimethoprime Sulfamethoxazole, Clindamycine, Neomycin, and Erythromycin than Coagulase-positive Staphylococci (CPS). CPS were more resis¬tant to Penicillin and Tetracycline as compared to CNS strains; however, all these strains presented sensitivity to Gentamicin and neomycin. Conclusion: The Staphylococci showed high resistance to the beta-lactam antibiotics. As far as the authors know, these molecules are used with or without control in different protocols to prevent and cure the mastitis in Algeria. [J Adv Vet Anim Res 2019; 6(2.000): 231-235]
Show more [+] Less [-]Alkaline treatment for preventing acidosis in the rumen culture fermenting carbohydrates: An experimental study in vitro
2019
Darwin | David Blignaut
Objective: The current research was carried out to evaluate the use of magnesium hydroxide as buffer to control acidosis in rumen culture fermenting carbohydrates in vitro. Materials and Methods: The experiments were carried out in the chemostat system in which the reactor used was a 200 ml of working volume. A series of fed-batch trials were carried out in fed-batch system with hydraulic retention time of 4 days. All digesters were completely mixed with the rotation of 55 rpm, and the temperature was controlled at 39°C ± 0.5°C. Results: Results showed that the supplementation of magnesium hydroxide (50 mM/day) to the corn starch feed (12.5 gm/l per day) for the rumen culture could prevent acidosis while at the same concentration of sodium bicarbonate addition to rumen culture, acidosis cannot be prevented in which lactic acid accumulated up to 200 mM. Supplementing magnesium hydroxide to the mixture of starch and sugar feeds prevented acidosis in which the major fermentation end product formed was acetate. A daily feeding with the ratio of 4.5:1 [starch: Mg(OH)2 ] was feasible to prevent rumen acidosis. Conclusion: Magnesium hydroxide added to the rumen culture could prevent lactic acid accumulation while sodium bicarbonate supplementation did not prevent acidosis and had lactic acid accumulation. [J Adv Vet Anim Res 2019; 6(1.000): 100-107]
Show more [+] Less [-]Vitamin E ameliorates testicular histological features and androgen binding protein levels in testicle of rats induced by allethrin
2019
Yofa Sukmawati | Dessy Arisanty | Alimuddin Tofrizal | Arni Amir
Objective: This study aims to determine the ameliorative effect of vitamin E (vit E) on histological features and androgen binding protein (ABP) levels in rats induced by allethrin. Materials and Methods: Thirty sexually mature male Wistar rats weighing between 200 and 300 gm, and aging 3 months were taken for this study and were divided into three groups: negative control (NC), positive control (PC), and treatment (T) groups. The PC and T groups were induced by allethrin 12 h per day for 31 days; however, only the T group was given vit E orally at 1 ml/gm body weight (BW) each day for 14 days. The paraffin block method was used to measure tubules diameter, thickness of the seminiferous epithelial layer, and Sertoli cell number. The ABP levels were measured by enzyme-linked immunosorbent assay. Results: The results showed that vit E gave significant effect (p < 0.05) on tubular diameter at NC 123.67 ± 12.77, PC 147.16 ± 10.64, and T 130.08 ± 10.00; tubular epithelial thickness at NC 33.55 ± 3.21, PC 30.02 ± 1.53, and T 32.96 ± 2.81; Sertoli cells number at NC 55.48 ± 5.9, PC 43.84 ± 3.77, and T 53.44 ± 4.26; and ABP levels at NC 72.35 ± 39.06, PC 38, 48 ± 18.78, and T 86.10 ± 35.77, respectively. Conclusion: This study concludes that vit E has an ameliorative effect against the toxic effects of allethrin at testicular histological features and ABP levels. [J Adv Vet Anim Res 2019; 6(4.000): 486-491]
Show more [+] Less [-]The effects of laser diode treatment on liver dysfunction of Mus musculus due to carbofuran exposure: An in vivo study
2019
Suryani Dyah Astuti | Vivi Sumanti Victory | Amalia Fitriana Mahmud | Alfian Pramudita Putra | Dwi Winarni
Objective: The aim of this study is to determine the effect of laser diode as an alternative treat¬ment on liver dysfunction (in vivo study) that is caused by carbofuran using male mice (Mus musculus) strain Balb/C. Materials and Methods: The samples were divided into three groups, namely, Group CL (con¬trol group, no treatment), Group C+L (only treated by carbofuran treatment), and Group C+L+ (treatment group, treated by carbofuran and laser-puncture) with five replications each. After being treated, each liver slice of samples was observed using microscope to get the histology result and then scored. Results: Carbofuran contamination can lead to inflammation of cells and necrosis. The histology results and the scoring test showed that the liver cells repair with the energy dose of laser diode at 0.5 and 1.0 Joule. Conclusion: The optimum energy dose in this study was 1.0 Joule which had the closest score of inflammatory cells and necrosis to normal liver cells. [J Adv Vet Anim Res 2019; 6(4.000): 499-505]
Show more [+] Less [-]Application of real-time polymerase chain reaction using species specific primer targeting on mitochondrial cytochrome-b gene for analysis of pork in meatball products
2019
Salmah Orbayinah | Hari Widada | Adam Hermawan | Sismindari Sudjadi | Abdul Rohman
Objective: This study aimed to design specific primers derived from mitochondrial cytb of Sus Scrofa (1F1R primer) used in the pork meatball analysis using real time polymerase chain reaction (RT-PCR) method. Materials and Methods: Such designed primers were validated and these included specificity of primer, linearity, and sensitivity of the method as well as the repeatability test. The primers were specifically affirmed in the fresh tissue of chickens, cows, pigs, and goats. The linearity and sensi¬tivity of the method was conducted by measuring the amplification curve from a series of dilution (0, 1, 1, 10, 100, 1,000, and 10,000 pg/μl of DNA) extracted from 100% pork meatball formulation. The repeatability test was conducted by determining the cycle threshold (Ct) values of RT-PCR amplification from 100% pork meatball formulation as many as six times. Results: Primer of 1F1R (forward: 5′-ACG CGA TAT AAG CAG GTA AA-3′; reverse: 5′-CTG CTT TCG TAG CAC GTA TT-3′) was specific in analyzing the presence of pork in meatball formulation at 47.1°C, which was optimum annealing temperature. The DNA identification was able to use the primers by RT-PCR with 1 pg as the limit of detection, efficiency value was 242.58%, and the coeffi¬cient of determination value (R2) was 0.956. The coefficient of variance was 4.13%. The developed method was also fruitfully applied to analyze commercial meatballs. Conclusion: RT-PCR method using specific primers targeting on mitochondrial gene (1F1R primer) could be used as the standard method for identification of pork in food samples intended for halal authentication studies. [J Adv Vet Anim Res 2019; 6(2.000): 260-265]
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