Standardization and characterization of adipose-derived stromal vascular fraction from New Zealand white rabbits for bone tissue engineering

Research (Published online: 25-02-2021)

24. Standardization and characterization of adipose-derived stromal vascular fraction from New Zealand white rabbits for bone tissue engineering

Khan Sharun, Abhijit M. Pawde, Rohit Kumar, E. Kalaiselvan, Prakash Kinjavdekar, Kuldeep Dhama and Amar Pal

Veterinary World, 14(2): 508-514

ABSTRACT

Background and Aim: Adipose tissue-derived stromal vascular fraction (SVF) contains a heterogeneous cell population comprising multipotent adipose-derived stem cells. Regenerative therapy using adipose-derived SVF has broad applications in bone tissue engineering due to the superior osteogenic potential of SVF. This study was designed to standardize and characterize adipose-derived SVF obtained from New Zealand white rabbits for bone tissue engineering and other potential applications.

Materials and Methods: Ten skeletally mature and clinically healthy adult New Zealand white rabbits were used in this study. The SVF was prepared using surgically resected interscapular adipose tissue following enzymatic digestion with 0.1% collagenase type I solution. The SVF pellet obtained after the final centrifugation step was suspended in a 0.5 mL control solution to obtain ready-to-use adipose-derived SVF. The freshly prepared SVF was characterized based on the total SVF cell count and cell yield per gram of adipose tissue. The SVF cells were enumerated using a hemocytometer.

Results: Interscapular adipose tissue depots are ideal for preparing autologous adipose-derived SVF due to the ease of access. The interscapular adipose-derived SVF prepared by enzymatic digestion had an average cell yield of 3.15±0.09×106 cells/g adipose tissue. Freshly prepared SVF had a total cell count of 3.15±0.09×104 cells/μL.

Conclusion: The enzymatic digestion of adipose tissue using 0.1% collagenase resulted in better cell yield per gram than methods previously reported in rabbits. The use of adipose-derived SVF can preclude the requirement for an additional culture period. In addition, it may also reduce the risk of extensive cell contamination, which makes it a safe and cost-effective strategy for bone tissue engineering applications.

Keywords: adipose tissue, characterization, New Zealand white rabbit, standardization, stromal vascular fraction.

Dietary effects of Garcinia kola seed meal on growth performance, hematology and serum biochemical parameters of weaned rabbits

Research (Published online: 25-02-2021)

23. Dietary effects of Garcinia kola seed meal on growth performance, hematology and serum biochemical parameters of weaned rabbits

Stanley Uzochukwu Ilo, Eunice Amaka Akuru, Jonathan Chinonso Egbo, Chika Ethelbert Oyeagu and Henry Oyeji Edeh

Veterinary World, 14(2): 499-507

ABSTRACT

Background and Aim: Garcinia kola seed (GKS) is used to prevent and cure a number of gastric-related ailments. GKS contain a copious amount of polyphenols and can be utilized as a natural growth promoter in the nutrition of weaned rabbits. This study aimed to determine the dietary effects of GKS meal (GKSM) on the growth performance, hematology and serum biochemical parameters of weaned rabbits.

Materials and Methods: GKS were dried and ground into powder. Thirty-two 8-week-old crossbred rabbits with an average weight of 614 g were randomly divided into four dietary groups. The diets were designated as follows: Control (corn-soybean based diet with 0% GKSM) and the GKSM-supplemented diets with 1.5% GKSM, 3% GKSM, and 4.5% GKSM. On the 56th day of the feeding trial, blood was collected from the marginal ear vein of all rabbits and transferred into two separate labeled tubes. The first set of blood was used to determine the hematological indices. The second set of blood was transferred into plain bottles and allowed to coagulate. The coagulated blood was subjected to standard methods of serum separation, and the sera were harvested and used to evaluate serum biochemical parameters.

Results: Although the average final body weight was highest in rabbits fed with 1.5% GKSM; this value was similar to rabbits fed with 0% and 4.5% GKSM. The average daily weight gain was highest in rabbits fed with 1.5% GKSM, while the feed conversion ratio was improved in the 0% and 1.5% GKSM groups. The dietary treatments also had a significant effect on the red blood cell count (RBC) and hemoglobin (Hb) concentration, while other blood parameters did not differ significantly (p>0.05). Higher inclusion levels (3% and 4.5%) of GKSM led to a significant increase in RBC and Hb values (p<0.05). The total protein increased at all levels of GKSM inclusion (p<0.05). Bilirubin, sodium, and potassium levels significantly decreased at 4.5% GKSM inclusion (p<0.05). Urea levels were lowered at 1.5% and 4.5% GKSM inclusion, while cholesterol levels were decreased at 3% and 4.5% dietary levels.

Conclusion: From the results of the present study, the supplementation of up to 4.5% GKSM revealed no harmful effect on the hematological and serum biochemical parameters of weaned rabbits, while their growth performance improved at a 1.5% inclusion level of GKSM.

Keywords: Garcinia kola, performance, production, rabbits, supplementation.

Radiographic lumbosacral vertebral abnormalities and constipation in cats

Research (Published online: 23-02-2021)

22. Radiographic lumbosacral vertebral abnormalities and constipation in cats

Chutimon Thanaboonnipat, Kamonchanok Kumjumroon, Kamonwan Boonkwang, Natthacha Tangsutthichai, Wassapon Sukserm and Nan Choisunirachon

Veterinary World, 14(2): 492-498

ABSTRACT

Background and Aim: Lumbosacral intervertebral disk disease (IVDD) in cats usually develops concurrent with constipation, spondylosis deformans, and sacralization. However, the prevalence of lumbar IVDD in cats was considered low, and there was less information on the incidence of non-traumatic lumbosacral vertebral abnormalities that may affect large bowel dysfunction. This study aimed to retrospectively investigate the relationship between non-traumatic lumbosacral vertebral abnormalities, both congenital and acquired, and large bowel dysfunction in cats.

Materials and Methods: Of 3108 cats that were presented to the Diagnostic Imaging Unit, the Small Animal Teaching Hospital, Faculty of Veterinary Science, Chulalongkorn University, between March 2016 and February 2018, 1365 cats met the inclusion criteria. All abdominal radiographs were reviewed, and all subsequent data were recorded, including the presence of congenital or acquired lumbosacral lesions, number of lumbar vertebrae, and length of the second, fifth, and last lumbar vertebrae, including the type of lumbar abnormalities. Moreover, radiographic information relating to constipation and megacolon was also collected.

Results: Non-traumatic lumbosacral vertebral abnormalities were observed in 29.74% of cats. The most common congenital lumbosacral vertebral abnormalities were six lumbar vertebrae, sacralization, and lumbarization, whereas most common acquired lumbosacral abnormalities were bone spur, narrowing disk space, spondylosis deformans, and lumbosacral degeneration, respectively. Cats with abnormal lumbosacral vertebrae are prone to have more problems with the large bowel (p=0.0057; odds ratio=1.731). Moreover, congenital and acquired lumbosacral abnormalities were also at risk of large bowel abnormalities (p=0.0069; odds ratio=1.920 and p<0.0001; odds ratio=4.107, respectively).

Conclusion: This study revealed the evidence and distribution of the variation in feline lumbar anatomy and also elucidated that cats with abnormal lumbar vertebral columns were more likely to have problems with distal gastrointestinal tracts than those without.

Keywords: cat, constipation, large bowel, lumbosacral, vertebrae.

Effect of peppermint oil and its microemulsion on necrotic enteritis in broiler chickens

Research (Published online: 23-02-2021)

21. Effect of peppermint oil and its microemulsion on necrotic enteritis in broiler chickens

Hend K. Sorour, Reham A. Hosny and Dalia M. A. Elmasry

Veterinary World, 14(2): 483-491

ABSTRACT

Background and Aim: Clostridium perfringens is one of the multiple drug-resistant intestinal pathogens causing necrotic enteritis disease, leading to great economic losses in poultry farms. This study aimed to evaluate the potential use of peppermint oil and its microemulsion (ME) as an alternative to antibiotics to control necrotic enteritis in broiler chickens.

Materials and Methods: Peppermint oil ME formulation (15% oil/water) was prepared and characterized by zeta potential, Fourier transform infrared, high-resolution transmission electron microscopy, and liquid chromatography–mass spectrometry (LC-MS/MS). The minimal inhibitory concentrations of the peppermint oil and its ME were investigated. A total of 80 commercial one day old Arbor Acres broiler chickens were randomly assigned to four groups of 20 birds each. The four groups were the negative control, positive control, peppermint oil (0.5 mL/mL water/10 days old), and its ME (0.25 mL/mL water/10 days old) groups. C. perfringens was orally provided at concentration of 1×108 CFU/mL on days 14, 15, and 16. Clinical signs and mortality were observed daily. Growth performance, gross lesions and cecal samples were investigated and examined on days 21, 28, and 35.

Results: Peppermint oil ME formulation has a polydispersity index, zeta potential and droplet size of 0.234, –24 mV±4.19, and 29.96±1.56 nm, respectively. LC–MS/MS analysis of oil and ME revealed common presence of phenolic compounds such as rosmorinic (360.31 g/mol), chlorogenic acid (354.31 g/mol), hesperidin (610.56 g/mol), and luteolin 7-O-β- glucuronide (462.1 g/mol). The treated groups with peppermint oil and ME showed lower lesions, mortality and colony-forming units in addition to higher growth performance (p<0.05) compared to the positive control group.

Conclusion: Our study suggests the potential efficacy of peppermint oil and ME in the reduction of necrotic enteritis lesions and C. perfringens count.

Keywords: broiler chicken, feed conversion ratio, microemulsion, nanotechnology, necrotic enteritis, peppermint oil.

Genetic variations in the Myostatin gene affecting growth traits in sheep

Research (Published online: 23-02-2021)

20. Genetic variations in the Myostatin gene affecting growth traits in sheep

Noha M. Osman, Heba I. Shafey, Mohamed A. Abdelhafez, Ahmed M. Sallam and Karima F. Mahrous

Veterinary World, 14(2): 475-482

ABSTRACT

Background and Aim: Sheep productivity in developing countries is crucial, as this animal is an essential source of meat and wool. Myostatin (MSTN) plays an important role in the regulation of muscle mass through the regulation of muscle growth, differentiation, and regeneration. The present study sought to investigate genetic variation in the first intron of the MSTN gene and the association of variants with growth traits in major sheep breeds in Egypt (Barki, Ossimi, and Rahmani) and Saudi Arabia (Najdi) using polymerase chain reaction (PCR) and sequencing.

Materials and Methods: Blood samples were collected, and DNA was extracted from 75 animals. A 386 bp fragment in the first intron of the MSTN gene was amplified using PCR. Polymorphic sites were detected using direct sequencing and then correlated with growth traits using a general linear model.

Results: Sequence analysis of the first intron of MSTN gene identified six single-nucleotide polymorphisms (SNPs) in the studied breeds. Four mutual SNPs were determined: c.18 G>T, c.241 T>C, c.243 G>A, and c.259 G>T. In addition, two SNPs c.159 A>T and c.173 T>G were monomorphic (AA and TT, respectively) in the Ossimi, Rahmani, and Najdi breeds and polymorphic in the Barki breed. The association analysis revealed that the c.18 G>T and c.241 C>T significantly associated (p<0.05) with birth weight and average daily weight gain, respectively.

Conclusion: Our results strongly support MSTN as a candidate gene for marker-assisted selection in sheep breeding programs. Furthermore, the identified variants may be considered as putative markers to improve growth traits in sheep.

Keywords: growth traits, Myostatin gene, polymorphism, sheep.

Investigation of Sarcocystis spp. in slaughtered cattle and sheep by peptic digestion and histological examination in Sulaimani Province, Iraq

Research (Published online: 22-02-2021)

19. Investigation of Sarcocystis spp. in slaughtered cattle and sheep by peptic digestion and histological examination in Sulaimani Province, Iraq

Shadan Hassan Abdullah

Veterinary World, 14(2): 468-474

ABSTRACT

Background and Aim: Sarcocystosis is a zoonotic infection caused by various species of Sarcocystis organisms with a worldwide geographic distribution. This study investigated the presence of Sarcocystis organisms in cattle and sheep slaughtered at an abattoir in Sulaimani Province in North Iraq.

Materials and Methods: A total of 130 muscle samples were collected during May, June, and July of 2020, including 80 samples from sheep and 50 samples from cattle. Samples were examined visually for macrosarcocysts. The peptic digestion method was used to analyze fresh muscle tissue samples for detecting microsarcocysts followed by microscopic examination. Furthermore, muscle samples were fixed and stained with hematoxylin and eosin for histopathological examination.

Results: In the gross examination, macroscopic cysts were not detected in both cattle and sheep; hence, all the prevalence data were obtained through microscopic observation of muscle samples. The peptic digestion method revealed the presence of banana-shaped bradyzoites in 90% and 92.5% of slaughtered cattle and sheep muscle samples, respectively. Organ-wise prevalence revealed that 95% and 92% of esophageal samples of sheep and cattle contained Sarcocystis spp., respectively. Moreover, 90% and 88% of sheep and cattle diaphragms were respectively infected. Histopathological examination of tissue sections revealed two morphologically distinct types of microsarcocysts, including thin-walled and thick-walled, in both sheep and cattle.

Conclusion: The suspected Sarcocystis spp. were Sarcocystis tenella and Sarcocystis arieticanis in sheep and Sarcocystis cruzi and Sarcocystis bovifelis or Sarcocystis hominis in cattle. Infective stages of different Sarcocystis spp. are widespread in the study area environment.

Keywords: cattle, muscle samples, prevalence rate, Sarcocystis, sheep.

Evaluation of the acute toxicity of the extracts of Anacyclus pyrethrum var. pyrethrum (L.) and Anacyclus pyrethrum var. depressus Maire in Swiss mice

Research (Published online: 22-02-2021)

18. Evaluation of the acute toxicity of the extracts of Anacyclus pyrethrum var. pyrethrum (L.) and Anacyclus pyrethrum var. depressus Maire in Swiss mice

Fatima Zahra Jawhari, Abdelfattah El Moussaoui, Hamada Imtara, Hamza Mechchate, Imane Es-Safi, Mohamed Bouhrim, Loubna Kharchoufa, Achraf Miry, Dalila Bousta and Amina Bari

Veterinary World, 14(2): 457-467

ABSTRACT

Background and Aim: Anacyclus pyrethrum (L.) has been used in traditional North African and Indian medicine for the treatment of several diseases such as cancer, rheumatism, epilepsy, diabetes, and Alzheimer's disease. Despite its medical benefits, few studies have examined its toxicity. The present study evaluated the acute toxicity of hydroethanolic extracts of different parts (roots, seeds, leaves, and capitula) of two varieties of A. pyrethrum (L.), namely, A. pyrethrum var. pyrethrum (L) and A. pyrethrum var. depressus (Ball) Maire, in mice.

Materials and Methods: Acute toxicity was evaluated after the oral administration of different extracts at doses of 300, 500, and 2000 mg/kg. Mortality, body weight, general behavior, and adverse effects were observed daily for 14 days. At the end of the experiment, mice were sacrificed, and biochemical parameters and histopathology of the liver, kidneys, and spleen were analyzed.

Results: The extracts of different parts of both plants induced no signs of toxicity or mortality during the observation period, excluding capitulum and seed extracts, which induced slight sedation at a dose of 2000 mg/kg. The LD50 of the extracts was estimated to exceed 2000 mg/kg. The administration of A. pyrethrum var. pyrethrum roots at a dose of 300 mg/kg resulted in significantly increased AST levels. However, the A. pyrethrum var. depressus root extract induced significant increases in the levels of both transaminases (alanine aminotransferase [ALT] and aspartate aminotransferase [AST]). The remaining extracts of both plants at a dose of 500 mg/kg significantly increased AST levels. Moreover, all plant extracts excluding the A. pyrethrum var. pyrethrum capitulum extract at 2000 mg/kg provoked significant increases in AST levels, and A. pyrethrum var. depressus roots provoked a significant increase of ALT levels. Meanwhile, mice treated with high doses of extracts (2000 mg/kg) displayed histopathological changes in the liver, kidneys, and spleen characterized by hepatic distress, inflammatory infiltration, focal tubular necrosis, vascular congestion, and lymphoid hyperplasia.

Conclusion: The results of the present study indicate that the hydroethanolic extracts of different parts of two varieties of A. pyrethrum (L.) were not toxic in mice at low concentrations, whereas some toxic effects were detected in mice treated at 2000 mg/kg.

Keywords: acute oral toxicity, Anacyclus pyrethrum var. depressus (Ball) Maire, Anacyclus pyrethrum var. pyrethrum (L), histopathology, serum biochemistry analysis.