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Nontyphoid Salmonella Prevalence, Serovar Distribution and Antimicrobial Resistance in Slaughter Sheep

  • Cetin, Ece;Temelli, Seran;Eyigor, Aysegul
    • Food Science of Animal Resources
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    • v.40 no.1
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    • pp.21-33
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    • 2020
  • This study aimed to determine the current prevalence, serovar distribution and antimicrobial resistance rate and patterns of nontyphoid Salmonella (NTS) in slaughter sheep and their edible offal. While filling the gap of up to date related information in Turkey, data presented is also of significance since contamination of ovine meat, its products and offal with this pathogen is threat to public health due to their considerably high consumption rates in our country. Current NTS carriage in 200 apparently healthy slaughter sheep by ISO 6579:2002, 6579:2002/A1:2007 standard bacteriology (ISO) was 5% (10/200) (4 fecal content - 2%, 3 mesenterial lymph node - 1.5%, 3 kidney - 1.5%) out of 1,400 samples (0.7%), with no isolation from carcass, liver, gallbladder, spleen. Real-time PCR was in substantial agreement to ISO in confirming Salmonella-suspect isolates (Relative Trueness: 93.6%). S. Newport (40%) was the predominant serovar, followed by the second prevalent serovars as S. Typhimurium and S. Kentucky (20%), and by S. Umbilo and S. Corvallis (10%). Four and 6 out of 10 NTS isolates were susceptible (40%) and resistant (60%) to 18 antimicrobials, respectively. S. Typhimurium isolates were multidrug resistant (MDR) to tigecycline and sulphamethoxazole/trimethoprim, with one also resistant to cefepime. S. Corvallis was MDR to ampicillin, ciprofloxacin, norfloxacin and pefloxacin. The predominance of S. Newport and first isolation of S. Corvallis in sheep in the world; first time isolations of Newport, Kentucky, Corvallis, Umbilo serovars from sheep in Turkey; and high antimicrobial resistance rates obtained in majority of the isolates highlights study findings.

Quantitative Evaluation of the First Order Creatine-Kinase Reaction Rate Constant in in vivo Shunted Ovine Heart Treated with Oxandrolone Using Magnetization Transfer 31P Magnetic Resonance Spectroscopy (MT-31P-MRS) and 1 H/31P Double-Tuned Surface Coil: a Preliminary Study

  • Thapa, Bijaya;Dahl, Marjanna;Kholmovski, Eugene;Burch, Phillip;Frank, Deborah;Jeong, Eun-Kee
    • Investigative Magnetic Resonance Imaging
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    • v.22 no.1
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    • pp.26-36
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    • 2018
  • Purpose: Children born with single ventricle physiology demonstrate poor growth rate and suffer from malnutrition, which lead to increased morbidity and mortality in this population. We assume that an anabolic steroid, oxandrolone, will promote growth in these infants by improving myocardial energy utilization. The purpose of this paper is to study the efficacy of oxandrolone on myocardial energy consumption in these infants. Materials and Methods: We modeled single ventricle physiology in a lamb by prenatally shunting the aorta to the pulmonary artery and then postnatally, we monitored cardiac energy utilization by quantitatively measuring the first order reaction rate constant, $k_f$ of the creatine-kinase reaction in the heart using magnetization transfer $^{31}P$ magnetic resonance spectroscopy, home built $^1H/^{31}P$ transmit/receive double tuned coil, and transmit/receive switch. We also performed cine MRI to study the structure and dynamic function of the myocardium and the left ventricular chamber. The spectroscopy data were processed using home-developed python software, while cine data were analyzed using Argus software. Results: We quantitatively measured both the first order reaction rate constant and ejection fraction in the control, shunted, and the oxandrolone-treated lambs. Both $k_f$ and ejection fraction were found to be more significantly reduced in the shunted lambs compared to the control lambs, and they are increased in oxandrolone-treated lambs. Conclusion: Some improvement was observed in both the first order reaction rate constant and ejection fraction for the lamb treated with oxandrolone in our preliminary study.

Comparison of Histopathology, Serology and PCR for the Diagnosis of Malignant Catarrhal Fever (Malignant Catarrhal Fever의 병리조직학적 진단과 혈청학적 진단 및 PCR 진단법의 비교)

  • Kim, Ok-jin;Crawford, Timothy B.
    • Korean Journal of Veterinary Research
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    • v.43 no.3
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    • pp.471-476
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    • 2003
  • Malignant catarrhal fever (MCF) is a systemic disease of ruminants caused by ovine herpesvirus 2 (OvHV-2). OvHV-2 is a gamma herpesvirus, which induces frequent latent infection and often difficult to detect its antigens and even specific nucleic acids because of its low viral copies in the infected tissues. Histopathology, serology and polymerase chain reaction (PCR) were compared for the diagnosis of MCF using 10 bison infected with OvHV-2. Histopathological diagnosis was performed using the criteria which was based upon the pathognomic lesions. Serological diagnosis was conducted using its serum with competitive ELISA for the detection of antibodies of OvHV-2. Also, the nest PCR was performed with peripheral blood leukocytes for the detection of OvHV-2-specific DNAs. Primers 556 and 775 were used for the primary amplification, and primers 556 and 555 were used for the secondary amplification. As the results, positive cases were 6 by histopahology, 9 by serology and 10 by PCR. As comparing with other diagnostic methods, PCR was found to be more sensitive than histopathology and serology. The recent development of molecular diagnostic assays has provided powerful tools for investigating how viruses survive in nature. Development of PCR specific for viruses has dramatically improved the accuracy of diagnosis of viruses in clinically infected animals. Furthermore, amplification of viral genomic material by nest PCR represents the most sensitive method for the detection of viruses and might be detected successfully even though very low viral DNA copies. So, it could be used as the first choice for the detection of viral DNAs with low copies such as the status of latent infection. However, it has also some limitation of application like as false negative results by PCR inhibitors and false positive results by contamination. The results of this study suggest that the use of molecular biological methods like PCR may increase the accuracy for the diagnosis of infectious diseases. However, in diagnostic laboratory, it is recommended that PCR assay must be conducted with other diagnostic methods for more reliable diagnosis.

Automatic Control System on Cardiac Output Regulation for the Moving Actuator Type Total Artificial Heart (MOVING-ACTUATOR TYPE 인공심장의 심박출 조절에 대한 자동 제어방법)

  • 김원곤
    • Journal of Chest Surgery
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    • v.28 no.6
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    • pp.542-548
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    • 1995
  • The goal of this study is to develop an effective control system for cardiac output regulation based upon the preload and afterload conditions without any transducers and compliance chambers in the moving actuator type total artificial heart. Motor current waveforms during the actuator movement are used as an input to the automatic control algorithm. While the current waveform analysis is performed, the stroke length and velocity of the actuator are gradually increased up to the maximum pump output level. If the diastolic filling rate of either right or left pump begins to exceed the venous return, atrial collapse will occur. Since the diastolic suction acts as a load to the motor, this critical condition can be detected by analyzing the motor current waveforms. Every time this detection criterion is met, the control algorithm decreases the stroke velocity and length of the actuator step by step just below the critical detection level. Then, they start to increase. In this way the maximum pump output under given venous return can be achieved. Additionally the control algorithm provides some degree of afterload sensitivity. If the aortic pressure is detected to exceed 120 mmHg, the stroke length and velocity decrease in the same way as the response to the preload. Left-right pump output balance is maintained by proper adjustment of the asymmetry of the stroke angle. In the mock circulatory test, this control system worked well and there was a considerable range of stroke volume difference with adjustment of the asymmetry value. Two ovine experiments were performed. It was confirmed that the required cardiac output regulation according to the venous return could be achieved with adequate detection of diastolic function, at least in the in vivo short-term survival cases[2-3 days . We conclude that this control algorithm is a promising method to regulate cardiac output in the moving actuator type total artificial heart.

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Preferential Peroxidase Activity of Prostaglandin Endoperoxide H Synthase for Lipid Peroxides

  • Yun, Seol-Ryung;Han, Su-Kyong;Song, In-Seok
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2001.11a
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    • pp.94-94
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    • 2001
  • Prostaglandin endoperoxide H synthase (PGHS) catalyzes the committed step in prostaglandins and thromboxane A$_2$-- oxygenation of arachidonic acid to the hydroperoxy endoperoxide PGG$_2$, followed by reduction PGG$_2$to the alcohol PGH$_2$. The two reactions by PGHS -- cyclooxygenase and peroxidase -- occur at distinct but structurally and functionally interconnected sites. The peroxidase reaction occurs at a heme-containing active site located near the protein surface. The cyclooxygenase reaction occurs in a hydrophobic channel in the core of the enzyme. Initially a peroxide reacts with the heme group, yielding Compound I and an alcohol derived from the oxidizing peroxide. Compound I next undergoes an intramolecular reduction by a single electron traveling from Tyr385 along the peptide chain to the proximal heme ligand, His388, and finally to the heme group. Following the binding of arachidonic acid, Tyr385 tyrosyl radical initiates the cyclooxygenase reaction by abstracting the 13-pro(5) hydrogen atom to give an arachidonyl radical, which sequentially reacts with two molecules of oxygen to yield PGG$_2$. In order to characterize PGHS peroxidase active site, we examined various lipid peroxides with purified recombinant ovine PGHS proteins and determined the rate constants. The results have shown that twenty-carbon unsaturated fatty acid hydroperoxides have similar efficiency in peroxidation by PGHS, irrespective of either the location of hydroperoxy group or the number of double bonds. It was also confirmed by the subsequent study with PGHS peroxidase active site mutants.

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Association between PCR-RFLP of Melatonin Receptor 1a Gene and High Prolificacy in Small Tail Han Sheep

  • Chu, M.X.;Ji, C.L.;Chen, G.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.12
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    • pp.1701-1704
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    • 2003
  • Melatonin regulates circadian rhythms and reproduction changes in seasonally reproductive mammals through binding to high-affinity, G-protein-coupled receptors. Small Tail Han sheep that has significant characteristics of high prolificacy and nonseasonal ovulatory activity is an excellent local sheep breed in P. R. China. The exon 2 of the ovine melatonin receptor 1a (MTNR1A) gene was amplified and a uniform fragment of 824 bp was obtained in 150 ewes of Small Tail Han sheep. The 824 bp PCR product was digested with restriction endonucleases Mnl I and Rsa I, and genetic polymorphism was detected by PCR-RFLP. Polymorphic Mnl I site was detected at base position 605 of the exon 2 of the MTNR1A gene. There were two kinds of genotypes in Small Tail Han sheep, AB (303 bp, 236 bp/67 bp) and BB (236 bp/67 bp, 236 bp/67 bp). The results indicated that genotype AA (303 bp, 303 bp) at Mnl I-RFLP site did not exist in non-seasonal estrous Small Tail Han sheep, which suggested that there was an association between genotype AA (303 bp, 303 bp) and reproductive seasonality in sheep. Polymorphic Rsa I site was detected at base position 604 of the exon 2 of the MTNR1A gene. Three kinds of genotypes were found in Small Tail Han sheep, AA (290 bp, 290 bp), AB (290 bp, 267 bp/23 bp) and BB (267 bp/23 bp, 267 bp/23 bp). Least squares means of litter size in the first parity and the second parity for genotype AA (290 bp, 290 bp) at Rsa I-RFLP site were 0.43 and 1.06 more than those for genotype AB (290 bp, 267 bp/23 bp) in Small Tail Han sheep.

Detection of copy number variation and selection signatures on the X chromosome in Chinese indigenous sheep with different types of tail

  • Zhu, Caiye;Li, Mingna;Qin, Shizhen;Zhao, Fuping;Fang, Suli
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.9
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    • pp.1378-1386
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    • 2020
  • Objective: Chinese indigenous sheep breeds can be classified into the following three categories by their tail morphology: fat-tailed, fat-rumped and thin-tailed sheep. The typical sheep breeds corresponding to fat-tailed, fat-rumped, and thin-tailed sheep are large-tailed Han, Altay, and Tibetan sheep, respectively. Detection of copy number variation (CNV) and selection signatures provides information on the genetic mechanisms underlying the phenotypic differences of the different sheep types. Methods: In this study, PennCNV software and F-statistics (FST) were implemented to detect CNV and selection signatures, respectively, on the X chromosome in three Chinese indigenous sheep breeds using ovine high-density 600K single nucleotide polymorphism arrays. Results: In large-tailed Han, Altay, and Tibetan sheep, respectively, a total of six, four and 22 CNV regions (CNVRs) with lengths of 1.23, 0.93, and 7.02 Mb were identified on the X chromosome. In addition, 49, 34, and 55 candidate selection regions with respective lengths of 27.49, 16.47, and 25.42 Mb were identified in large-tailed Han, Altay, and Tibetan sheep, respectively. The bioinformatics analysis results indicated several genes in these regions were associated with fat, including dehydrogenase/reductase X-linked, calcium voltage-gated channel subunit alpha1 F, and patatin like phospholipase domain containing 4. In addition, three other genes were identified from this analysis: the family with sequence similarity 58 member A gene was associated with energy metabolism, the serine/arginine-rich protein specific kinase 3 gene was associated with skeletal muscle development, and the interleukin 2 receptor subunit gamma gene was associated with the immune system. Conclusion: The results of this study indicated CNVRs and selection regions on the X chromosome of Chinese indigenous sheep contained several genes associated with various heritable traits.

Expression of pituitary adenylate cyclase activating polypeptide in the adult rat testis by in situ hybridization and immunohistochemistry (In situ hybridization법과 면역조직화학적법을 이용한 성숙한 흰쥐고환에서의 pituitary adenylate cyclase activating polypeptide의 발현)

  • Koh, Phil-ok;Kwak, Soo-dong
    • Korean Journal of Veterinary Research
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    • v.41 no.1
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    • pp.1-6
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    • 2001
  • Pituitary adenyl ate cyclase activating polypeptide (PACAP) was originally isolated from the ovine hypothalamus and stimulated cAMP production in anterior pituitary cells. It is known that PACAP stimulates cAMP accumulation and contributes to the spermatogenesis and steroidogenesis in rat testis. The principal aim of this study is to determinate the distribution of PACAP mRNA and protein in adult rat testis. For this study, we used in situ hybridization and immunohistochemistry techniques in adult rat testis. PACAP mRNA was stage specifically expressed in seminiferous tubules. Positive signals of PACAP mRNA were detected in the developing germ cells at stages HI-VII of the epithelial cycle. The strongest signals of PACAP mRNA and protein were detected in round spermatids at stages V to early VII of the cycle. These results demonstrate that PACAP which is synthesised in the developing germ cells contributes to the spermatogenesis in rat testis. Thus, we suggest that PACAP plays a critical role in the function of testis.

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Thymosin Beta4 Regulates Cardiac Valve Formation Via Endothelial-Mesenchymal Transformation in Zebrafish Embryos

  • Shin, Sun-Hye;Lee, Sangkyu;Bae, Jong-Sup;Jee, Jun-Goo;Cha, Hee-Jae;Lee, You Mie
    • Molecules and Cells
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    • v.37 no.4
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    • pp.330-336
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    • 2014
  • Thymosin beta4 (TB4) has multiple functions in cellular response in processes as diverse as embryonic organ development and the pathogeneses of disease, especially those associated with cardiac coronary vessels. However, the specific roles played by TB4 during heart valve development in vertebrates are largely unknown. Here, we identified a novel function of TB4 in endothelial-mesenchymal transformation (EMT) in cardiac valve endocardial cushions in zebrafish. The expressions of thymosin family members in developing zebrafish embryos were determined by whole mount in situ hybridization. Of the thymosin family members only zTB4 was expressed in the developing heart region. Cardiac valve development at 48 h post fertilization was defected in zebrafish TB4 (zTB4) morpholino-injected embryos (morphants). In zTB4 morphants, abnormal linear heart tube development was observed. The expressions of bone morphogenetic protein (BMP) 4, notch1b, and hyaluronic acid synthase (HAS) 2 genes were also markedly reduced in atrio-ventricular canal (AVC). Endocardial cells in the AVC region were stained with anti-Zn5 antibody reactive against Dm-grasp (an EMT marker) to observe EMT in developing cardiac valves in zTB4 morphants. EMT marker expression in valve endothelial cells was confirmed after transfection with TB4 siRNA in the presence of transforming growth factor ${\beta}$ ($TGF{\beta}$) by RT-PCR and immunofluorescent assay. Zn5-positive endocardial AVC cells were not observed in zTB4 morphants, and knockdown of TB4 suppressed TGF-${\beta}$-induced EMT in ovine valve endothelial cells. Taken together, our results demonstrate that TB4 plays a pivotal role in cardiac valve formation by increasing EMT.

Application of the Total Artificial Heart as an Implantable Biventricular Assist Device by Left Thoracotomy in an Ovine Model (양에서 좌측 개흉술 하에 완전인공심장의 체내이식형 양심실 보조장치로 사용에 관한 연구)

  • 원태희;민병구;김원곤
    • Journal of Chest Surgery
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    • v.34 no.4
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    • pp.296-304
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    • 2001
  • 배경: 이번 연구의 목적은 체내이식형 양심실 보조장치의 개발을 위한 안전하고 간편한 동물실험 모델을 확립하는 것이며 이동작동형 완전 인공심장의 양심실 보조장치로의 사용 가능성을 알아보는 것이다. 대상 및 방법: 7마리의 Corridale 양을 대상으로 실험하였다. 5번째 늑간극을 통하여 좌측 개흉술을 시행하고 좌측 유출로 캐뉼라는 하행 대동맥에 Dacron graft을 사용하여 연결하였고 유입로 캐뉼라는 좌심방에 삽입하였다. 우측 유출로 캐뉼라는 하행 대동맥과 같은 방법으로 주폐동맥에 연결하였으며 유입로 캐뉼라는 혜동맥을 하방으로 젖혀 우심방이를 노출시킨 다음 우심방이에 삽입하였다. 4마리에 있어서는 양심실 보조장치를 전복막강(preperitoneal space)에 위치시켰으며 2마리에 촤측흉강 내에, 1마리에서는 외부에 위치시켰다. aPTT는 수술전 aPTT의 2∼2.5배로 맞추어 주었다 결과: 수술중 또는 수술직후 사망한 경우는 없었으며 7마리 모두 인공호흡기 이탈이 가능하였고 스스로 기립하고 음식물 섭취도 가능하였다. 1마리는 술후 2일째 혈전 색전증 및 인공호흡기 작동 잘못으로 사망하였으며 2마리의 겨우는 술후 2일째 각각 양심실 보조장치의 작동 잘못 및 호흡부전으로 사망하였고, 3마리의 경우는 술후 4일째 혈전색전증으로 인한 급성신부전 및 호흡부전 등으로 사망하였다. 1마리에 있어서는 28일간 생존하였으며 양심실 보조장치의 구동 장치에 연결된 라인의 마로로 인해 양심실 보조장치의 작동이 정지되었으나 바로 양심실 보조장치를 제거하고 살릴 수 있었다. 모든 경우에 있는 캐뉼라를 삽입하고 양심실 보조장치를 작동시키는데 큰 어려움이 없었다. 7마리 중 4마리에서 혈전이 발견되었으며 특히 판막을 연결한 부위에 혈전생성이 많았다. 그러나 캐뉼라에는 혈전생성이 없었다. 결론: 양에 있어서 좌측 개흉술 및 전복막강 공간에 양심실 보조장치를 위치시키는 이번 동물실험 모델은 체내 이식형 양심실 보조장치 개발을 위한 좋은 동물실험 모델이라고 생각된다. 또한 디동 작동기형 완전 인공심장의 혈전생성을 억제할수 있는 방법들의 연구개발이 지속된다면 장기간의 체내 이식형 양심실 보조장치로 사용할 수 있을 것으로 생각된다.

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