Effect of low oxygen tension on transcriptional factor OCT4 and SOX2 expression in New Zealand rabbit bone marrow-derived mesenchymal stem cells

Research (Published online: 18-11-2020)

25. Effect of low oxygen tension on transcriptional factor OCT4 and SOX2 expression in New Zealand rabbit bone marrow-derived mesenchymal stem cells

Erma Safitri

Veterinary World, 13(11): 2469-2476

ABSTRACT

Background and Aim: Octamer-binding transcription factor 4 (OCT4) and sex-determining region Y-box 2 (SOX2) are transcription factors whose functions are essential to maintain the pluripotency of embryonic stem cells. The purpose of this study was to derive stem cells for in vitro culture and to maintain their viability and pluripotency, with the goal to obtain a cell line for transplantation in patients with degenerative diseases or injuries. This research focused on examining the effect of low oxygen tension on the ability of bone marrow-derived mesenchymal stem cells (BM-MSCs) to express OCT4 and SOX2 in vitro.

Materials and Methods: BM-MSCs were obtained from femurs of 2000 to 3000 g New Zealand male rabbits. BM-MSCs were divided into three groups to test different culture conditions: A control group under hyperoxia condition (21% O2) and two treatment groups with low oxygen tension (1% and 3% O2). We characterized the BM-MSCs using flow cytometric measurement of cluster differentiation 44 (CD44) and cluster differentiation 90 (CD90) expression. The expression of OCT4 and SOX2 was measured by immunofluorescence staining after 48 h of incubation in chambers with normal or low oxygen tension with controlled internal atmosphere consisting of 95% N2, 5% CO2, and 1% O2 (T1) and 3% O2 (T2). We considered OCT4 and SOX2 as two markers of pluripotency induction. All immunofluorescence data were subjected to a post hoc normality Tukey's honestly significant difference test; all differences with p<5% were considered significant.

Results: BM-MSCs were positive for CD44 and CD90 expression after isolation. Oxygen tension culture conditions of 1% and 3% O2 led to OCT4 and SOX2 expression on culture days 2 and 4 (p<0.05), respectively, as compared to the hyperoxia condition (21% O2).

Conclusion: Based on the OCT4 and SOX2 immunofluorescence data, we conclude that the stem cells were pluripotent at low O2 tension (at 1% O2 on day 2 and at 3% O2 on day 4), whereas under 21% O2 the OCT4 and SOX2 were not expressed.

Keywords: bone marrow, cluster differentiation 44, cluster differentiation 90, culture in vitro, low O2 tension, mesenchymal stem cells, octamer-binding transcription factor 4, pluripotency, sex-determining region Y-box 2.

Characterization of Burkholderia pseudomallei from spontaneous melioidosis in a Bornean orangutan

Research (Published online: 18-11-2020)

24. Characterization of Burkholderia pseudomallei from spontaneous melioidosis in a Bornean orangutan

Vincentius Arca Testamenti, Maryati Surya, Uus Saepuloh, Diah Iskandriati, Maryos Vigouri Tandang, Lia Kristina, Aris Tri Wahyudi, Dondin Sajuthi, Vivi Dwi Santi, Fiet Hayu Patispathika, Muhtadin Wahyu, Anton Nurcahyo and Joko Pamungkas

Veterinary World, 13(11): 2459-2468

ABSTRACT

Background and Aim: Melioidosis is a potentially fatal disease affecting humans and a wide range of animal species; it is often underdiagnosed and underreported in veterinary medicine in Indonesia. This study aimed to characterize morphological and molecular features of Burkholderia pseudomallei, the causative agent of melioidosis which caused the death of a Bornean orangutan.

Materials and Methods: Pulmonary abscess samples were cultured on several types of media, including Ashdown agar, Ashdown broth, and MacConkey agar. Type three secretion system orf 2 real-time polymerase chain reaction (PCR) and latex agglutination tests were performed to identify the bacteria. Morphological characteristics were compared to all previously published morphotypes. Subsequently, the bacteria were characterized by multilocus sequence typing (MLST) and Yersinia-like flagellum/Burkholderia thailandensis-like flagellum and chemotaxis PCR. The results of the genotyping were afterward compared to all genotypes from Southeast Asia.

Results: Multiple morphotypes of B. pseudomallei were perceived during the growth on Ashdown agar. Furthermore, it was identified by MLST that the Type I and Type II morphotypes observed in this study were clones of a single ST, ST54, which is predominantly found in humans and the environment in Malaysia and Thailand, although a very limited number of reports was published in association with animals. Moreover, the E-BURST analysis showed that the ST is grouped together with isolates from Southeast Asian countries, including Malaysia, Thailand, Singapore, and Cambodia. ST54 was predicted to be the founding genotype of several STs from those regions.

Conclusion: B. pseudomallei ST54 that caused the death of a Bornean orangutan has a distant genetic relationship with other STs which were previously reported in Indonesia, implying a vast genetic diversity in Indonesia that has not been discovered yet.

Keywords: Burkholderia pseudomallei, melioidosis, molecular characterization, nonhuman primate, orangutan.

Efficacy of novel antiseptic product containing essential oil of Lippia origanoides to reduce intramammary infections in cows

Research (Published online: 17-11-2020)

23. Efficacy of novel antiseptic product containing essential oil of Lippia origanoides to reduce intramammary infections in cows

Natalia Arantes Marcelo, Viviane Aguiar Andrade, Cintya Neves Souza, Rodrigo Pereira Mourão, Mário Henrique França Mourthe, Lívia Mara Vitorino Silva, Alessandra Rejane Ericsson de Oliveira Xavier, Mauro Aparecido de Sousa Xavier, André Augusto Gomes Faraco and Anna Christina Almeida

Veterinary World, 13(11): 2452-2458

ABSTRACT

Background and Aim: The use of antimicrobials in the control of mastitis is of concern in public health due to their inefficiency in targeting microorganisms. Studies with medicinal plants have risen as an alternative to the use of conventional products. The objective of this study was to evaluate the efficacy of an experimental disinfectant based on the essential oil (EO) from Lippia origanoides in preventing the development of new intramammary infections (IMI) in Holstein cows.

Materials and Methods: The conventional protocol of pre- and post-milking was used and the control (Conventional treatment [CNV]) and experimental (Experimental treatment [PEX]) products containing EO at 120 μL/mL were applied by immersion. Individual milk samples were analyzed using sheep blood agar methodologies and biochemical tests. The efficiency of the treatment was defined by the presence or absence of Staphylococcus aureus, coagulase-negative Staphylococcus, and Streptococcus spp.

Results: There were no clinical and subclinical mastitis cases, no lesions in the mucosal of teats, nor dirt score between groups in this study. Both treatments did not influence the occurrence of IMI.

Conclusion: The results revealed that PEX acts efficiently against microorganisms compared to the disinfection by the conventional product demonstrating the efficacy of the alternative product on the prevention of new IMIs in dairy cows.

Keywords: Intramammary infections, Lippia origanoides, mastitis, pre- and post-milking, teat disinfectant.

Ultrasonographic evaluation of the effects of the administration of neostigmine and metoclopramide on duodenal, cecal, and colonic contractility in Arabian horses: A comparative study

Research (Published online: 16-11-2020)

22. Ultrasonographic evaluation of the effects of the administration of neostigmine and metoclopramide on duodenal, cecal, and colonic contractility in Arabian horses: A comparative study

Noha Abdallah Beder, Ahmed Atef Mourad and Mahmoud Allam Aly

Veterinary World, 13(11): 2447-2451

ABSTRACT

Background and Aim: As means for enhancing gastrointestinal (GI) motility, prokinetics have previously been evaluated in different breeds of horses with varying success. No previous studies were conducted to evaluate the effect of prokinetics on the intestinal tract of Arabian horses breed. Using B-mode ultrasound in the quantification of intestinal contractility, this study aimed to compare the impact of neostigmine and metoclopramide on the intestinal motility of healthy Arabian horses.

Materials and Methods: Twenty-one clinically healthy Arabian horses were equally distributed into three groups. The control group was administered with 5 mL normal saline intramuscularly (IM). The second group was administered with neostigmine (0.044 mg/kg body weight [BW], IM), and the third group was administered with metoclopramide (0.25 mg/kg BW, IM). Duodenal, cecal, and colonic contractions were counted through ultrasonography for 3 min (pre-administration and 15, 30, 60, 120, and 180 min post-administration).

Results: In the neostigmine group, a significant (p<0.05) increase in duodenal, cecal, and colonic contractions was observed 15 min post-administration compared with that in the control group (15±1.0, 11.33±1.53, and 12.33±2.31 vs. 11.0±2.0, 6.33±0.58, and 5.33±0.58 contractions per 3 min, respectively), continuing to 60 min post-administration for the duodenum and cecum and 120 min for colon and then returning to the normal; however, the metoclopramide group showed a significant (p<0.05) increase only in cecal and colonic contractions 60 min post-administration compared with the control group (11.0±1.0 and 12.33±0.58 vs. 6±1.0 and 5.67±0.58 contractions per 3 min, respectively), continuing until the end of the experiment. Excessive sweating, excitation, and straining were recorded following the administration of neostigmine, whereas no side effects were observed in the metoclopramide group.

Conclusion: Neostigmine improves duodenal, cecal, and colonic contractions in healthy adult Arabian horses, whereas metoclopramide only improves cecal and colonic contractions. Metoclopramide appears to be safer and longer acting than neostigmine in the Arabian horse breed. Ultrasonography is a valuable noninvasive tool for the quantification of intestinal contractility. Future studies should consider the use of various dosages of metoclopramide and administration routes and investigate its impact on horses with GI transit disorders and inclusion in colic post-operative care.

Keywords: Arabian horse, intestinal contractility, metoclopramide, neostigmine, ultrasonography.

Effect of follicle-stimulating hormone on Bligon goat oocyte maturation and embryonic development post in vitro fertilization

Research (Published online: 14-11-2020)

21. Effect of follicle-stimulating hormone on Bligon goat oocyte maturation and embryonic development post in vitro fertilization

Diah Tri Widayati and Mulyoto Pangestu

Veterinary World, 13(11): 2443-2446

ABSTRACT

Background and Aim: Bligon goat is a crossbreed between Etawah and Kacang goat. This crossbreed goat is mostly reared by small farmers. In vitro maturation allows female goat (does) contributes toward reproduction despite the fact that the animal has been slaughtered. The aim of this study was to determine the in vitro maturation rate of Bligon goat oocytes supplemented with follicle-stimulating hormone (FSH), and their ability for further embryonic development after in vitro fertilization.

Materials and Methods: Experiment was conducted at the Laboratory of Animal Physiology and Reproduction, Faculty of Animal Science, Universitas Gadjah Mada, Yogyakarta, using Bligon goat ovaries obtained from local slaughterhouse around Yogyakarta. One thousand five hundred cumulus-oocyte complexes were matured for 24 h in tissue culture medium 199 supplemented with 50 IU/L FSH or without FSH (control). First, matured oocytes were evaluated its morphology based on the expansion of cumulus cells and PB1 extrusion. Next, 600 oocytes were then stained with 1% aceto-orcein to examine maturation based on changes in the configuration of chromosomes and nuclear membrane breakdown. Oocytes were considered mature when they reached metaphase II. To prove the ability of mature oocytes to develop into embryos, 900 oocytes were processed for fertilization in vitro. The data were analyzed using analysis of variance.

Results: The results indicated that FSH supplementation significantly increased oocyte maturation rate (65.21±7.26 vs. 43.25±6.23%) as indicated by extrusion of PB1 and homologous chromosome pairing and lined in the equator. The rate of degeneration was lower in the FSH-supplemented medium (3.21±0.25 vs. 10.17±3.15%). The blastocyst stage of oocyte developed embryos was reached by 12.43±2.15% and 22.28±4.86% of the control and treatment groups, respectively.

Conclusion: FSH supplementation significantly improves oocyte maturation and yields mature oocytes for future embryo development in vitro.

Keywords: Bligon goat, follicle-stimulating hormone supplementation, in vitro embryo production.

Left ventricular systolic function in dogs with pulmonic stenosis

Research (Published online: 14-11-2020)

20. Left ventricular systolic function in dogs with pulmonic stenosis

Ploypanut Trikhun, Sirilak Disatian Surachetpong, Saikaew Sutayatram and Chollada Buranakarl

Veterinary World, 13(11): 2436-2442

ABSTRACT

Background and Aim: Pulmonic stenosis (PS) is the most common congenital heart disease in dogs. This condition causes right ventricle (RV) overload and disrupts overall systolic function. The aim of this study was to examine the alterations of cardiac electrical activity and mechanical function in dogs with PS compared to normal healthy dogs.

Materials and Methods: The ventricular systolic function of dogs with PS was studied. Dogs were divided into two groups, PS (n=13) and control (CONT) (n=12). Measurements of blood pressure, electrocardiography (ECG), and echocardiography were performed.

Results: PS dogs had exercise intolerance, with six experiencing syncope. ECG of PS dogs showed higher amplitudes of P, S, and T waves (p<0.01), and a lower R:S ratio (p<0.001) with longer QRS duration (p<0.001) compared to CONT dogs. Echocardiography demonstrated that the pulmonic flow velocity and pressure gradient (PG) between the RV and the pulmonary artery of PS dogs were significantly higher than CONT dogs (p<0.001). The RV free wall thickness to the left ventricular posterior wall thickness ratio and the right atrium to the left atrium diameter ratio was higher (p<0.001), while interventricular septum (IVS) was thicker (p<0.01) in PS dogs compared with CONT dogs (p<0.001). The systolic function in PS dogs showed higher pulmonic valve velocity time integral (PVVTI) value (p<0.001) and longer pulmonic valve ejection time (ET) (p<0.05) than CONT dogs. However, aortic valve VTI (AVVTI) value and aortic valve ET were not significantly different between the groups, although fractional shortening in PS dogs was higher. In PS dogs, PG showed a significant positive correlation with PVVTI:AVVTI ratio (p<0.05).

Conclusion: PS had prolonged pulmonic valve opening. The thickening of the RV wall and IVS can cause a detrimental reduction of the left ventricular preload in PS dogs.

Keywords: dogs, pulmonic stenosis, ventricular function.

Application of statistical process control for monitoring bulk tank milk somatic cell count of smallholder dairy farms

Research (Published online: 13-11-2020)

19. Application of statistical process control for monitoring bulk tank milk somatic cell count of smallholder dairy farms

Veerasak Punyapornwithaya, Chalutwan Sansamur, Tawatchai Singhla and Paramintra Vinitchaikul

Veterinary World, 13(11): 2429-2435

ABSTRACT

Background and Aim: Consistency in producing raw milk with less variation in bulk tank milk somatic cell count (BMSCC) is important for dairy farmers as their profit is highly affected by it in the long run. Statistical process control (SPC) is widely used for monitoring and detecting variations in an industrial process. Published reports on the application of the SPC method to smallholder farm data are very limited. Thus, the purpose of this study was to assess the capability of the SPC method for monitoring the variation of BMSCC levels in milk samples collected from smallholder dairy farms.

Materials and Methods: Bulk tank milk samples (n=1302) from 31 farms were collected 3 times/month for 14 consecutive months. The samples were analyzed to determine the BMSCC levels. The SPC charts, including the individual chart (I-chart) and the moving range chart (MR-chart), were created to determine the BMSCC variations, out of control points, and process signals for each farm every month. The interpretation of the SPC charts was reported to dairy cooperative authorities and veterinarians.

Results: Based on a set of BMSCC values as well as their variance from SPC charts, a series of BMSCC data could be classified into different scenarios, including farms with high BMSCC values but with low variations or farms with low BMSCC values and variations. Out of control points and signals or alarms corresponding to the SPC rules, such as trend and shift signals, were observed in some of the selected farms. The information from SPC charts was used by authorities and veterinarians to communicate with dairy farmers to monitor and control BMSCC for each farm.

Conclusion: This study showed that the SPC method can be used to monitor the variation of BMSCC in milk sampled from smallholder farms. Moreover, information obtained from the SPC charts can serve as a guideline for dairy farmers, dairy cooperative boards, and veterinarians to manage somatic cell counts in bulk tanks from smallholder dairy farms.

Keywords: bulk milk somatic cell count, control chart, dairy farm, smallholder, statistical process control.