臺灣博碩士論文加值系統

第一部分

現今被廣泛飼養作為寵物兔的家兔（Oryctolagus cuniculus），乃由許多不同的品種組成，其中寵物兔族群是由各種純種和雜交的品種所組成。然而，在兔子所使用的標準值（Reference Intervals; RIs）由於過時方法和單一品種的樣本族群，致使其應用至寵物兔族群是有限制的。本研究旨在建立一個更完整、更新的紅細胞以及血清生化RIs，供混種寵物兔族群使用，以改善常規臨床診斷的準確性，並探討混種在紅細胞和生化參數的影響。本研究取樣自85隻臨床上健康的混種兔，並根據美國獸醫臨床病理學會之大綱建議，建立紅血球指數及12種血清生化RIs，並將本研究結果與四種廣泛使用的RIs進行比較，統計分析年齡及性別之差異。本研究RIs顯示與紐西蘭大白兔（New Zealand White Rabbits; NZWs）明顯不同，表示使用NZWs的RIs並非都適用於臨床寵物兔之病患。品種相關的效應可在天門冬氨酸轉移酶、鹼性磷酸酶和葡萄糖明顯顯現，而混種相關的效應則主要顯現於總蛋白、白蛋白、尿素氮、肌酸酐、鈣和磷酸鹽。具有統計上年齡差異同時具有臨床相關性的指數包括血紅素、血容比、白血球總數及肌酸酐。對於專門治療由不同品系和年齡所組成的寵物兔之臨床獸醫而言，建立專門針對混種寵物兔的RIs將有助其進行臨床更精確的判讀。特定年齡族群的RIs在未來有進一步調查建立之必要。


第二部分

腎臟病(Kidney Disease; KD)是寵物兔常見的老年疾病，而肌酸酐是特寵臨床實踐中最常使用的KD生物標記，可惜其敏感度受到許多因子之影響，導致實用性不足。其他的腎臟生物標記，如尿比重(Urine Specific Gravity; USG)、尿蛋白與尿肌酸酐比例(Urine protein to creatinine ratio; UPC ratio)和尿液ϒ-谷氨酰轉移酶(Urinary ϒ-glutamyl transferase; uGGT)濃度，均可以更早發現腎功能的減退，這些年來在狗貓已被充分的研究；然而，卻很少有研究調查這些生物標記於寵物兔的實用性，在臨床上也因為缺乏這些指數的標準值而少有獸醫去使用。本研究旨在建立UPC ratio、USG以及uGGT的標準值，並探討這些尿液參數是否能作為偵測寵物兔腎臟相關異常的臨床指標。本研究一共納入了30隻健康兔及11隻KD兔。結果顯示，混種寵物兔尿液蛋白的標準值顯著低於以純種寵物兔為主之文獻結果，此差異可能與品種相關的變異性有關。此外，KD組的肌酸酐和UPC ratio顯著高而USG顯著低於健康組。最後，接收者操作特徵曲線分析（Receiver operator characteristic curve; ROC curve）結果顯示，UPC ratio、USG以及uGGT均具有高的曲線下面積（Area under curve; AUC），可視為有良好的偵測表現。而將肌酸酐合併使用以上尿液生物標記的AUC，顯示出較個別使用更好的診斷表現。總結以上，本研究指出尿液參數於混種寵物兔使用的臨床價值，並且這些尿液參數有潛力成為用以偵測寵物兔腎臟相關異常的重要生物指標，本研究存在一些限制，如樣本數較小，且所包含的病例多為慢性腎臟病(chronic kidney disease ; CKD)，因此這些生物指標的診斷表現，值得未來進一步研究。

Part I

Domestic rabbits (Oryctolagus cuniculus) have numerous breeds, and the pet rabbit population consists of a variety of pure- and cross-breeds. However, the RIs for rabbits are limited by outdated methods and a single-breed source. The study aimed to establish more complete, updated erythrocytic and biochemical RIs for the cross-breed pet rabbit population to improve routine diagnostic accuracy in clinical practice, and to examine the cross-breed effects on erythrocytic and biochemical indices. Blood specimens were collected from 85 clinically healthy cross-breed pet rabbits representing a wide range of breeds. Reference Intervals for the hematological and 12 chemistry analytes were determined as recommended by the American Society for Veterinary Clinical Pathology guidelines. The results were compared to 4 widely used reference RIs. Age and sex differences for all parameters were statistically analyzed. The results diverged from the RIs for New Zealand White rabbits, indicating that not all RIs from this species are appropriate for daily clinical exotic animal practice. The breed-associated variations were evident for aspartate aminotransferase, alkaline phosphatase, and glucose while cross-breed effects were found for creatinine, blood urea nitrogen, total protein, albumin, phosphate, and calcium. Statistically significant differences and clinical relevance among age groups were detected for hemoglobin, hematocrit, total white blood cell count and creatinine. Establishing RIs specifically for cross-breed pet rabbits is desirable for clinicians treating pet rabbits of various breeds and ages. Age-specific RIs for this population warrants further investigation.


Part II

Kidney disease (KD) is a common geriatric disorders in pet rabbits. On the daily clinical practice, creatinine is the most commonly used biomarkers of KD but is influenced by many factors therefore not sensitive. Additional renal biomarkers, such as USG, UPC ratio, and uGGT, which allow early detection of decreased kidney function, are well-studied in dogs and cats1, however, few researches investigated the utility of these biomarkers in pet rabbits. In addition, there was no appropriate RIs for these parameters therefore they were rarely put into practice by veterinarians in the clinic. The present study aimed to establish the RIs for UPC ratio, USG and uGGT index and to explore the applicability of these urinary indices as indicators of KD in pet rabbits. Thirty healthy rabbits and 11 rabbits with KD were included. The comparative results showed significantly lower RI for urine protein in cross-breed pet rabbits compared with the literature mainly consist of pure-breed rabbits, the disparity may due to the breed-associated differences. In addition, significantly higher CRE and UPC ratio, lower USG were found in KD group compared to the healthy group. Finally, the receiver operator characteristic curve analysis showed high AUC (>0.75) of UPC ratio, USG and uGGT index, indicating good performances. The combined use of CRE with UPC ratio (0.991), and CRE with uGGT index (0.967) is higher than that of individual CRE (0.955), UPC ratio (0.934) or uGGT index (0.730), suggests even better performance. To conclude, the current study highlights the clinical value of the urinary indices in cross-breed pet rabbit population. However, some limitations existed such as relatively small sample size and the included cases were mostly CKD, thus the performances of these biomarkers worth further investigating.

第一部分 摘要 i
Part I Abstract ii
第二部分 摘要 iii
Part II Abstract iv
Tables of contents v
List of tables vii
List of figures viii
List of abbreviation ix
Part I 1
Chapter 1、 Introduction 2
Chapter 2、 Literature review 3
1. Domestic rabbits (Oryctolagus cuniculus) 3
2. Reference intervals 3
Chapter 3、 Materials and Methods 10
1. Animals selection and study design 10
2. Sample collection and processing 10
3. Analytes 11
4. Statistical Analysis 12
Chapter 4、 Results 14
1. Population 14
2. The comparative results 14
3. Effect of sex 14
4. Effect of age 15
Chapter 5、 Discussion 16
1. The importance of the study 16
2. The comparative results with source 1 and 2 16
3. The comparative results with source 4 17
4. Breed-associated variations 17
5. Cross-breed effect 19
6. Effect of sex 20
7. Effect of age 20
8. Conclusion and limitations 21
Tables 24
Figures 34
Part II 37
Chapter 1、 Introduction 38
Chapter 2、 Literature review 39
1. Application of creatinine in kidney disease 39
2. Application of urinalysis in kidney disease 40
3. Application of Urine Specific Gravity in kidney disease 43
4. Application of proteinuria in kidney disease 43
5. Application of urinary ϒ-GT in kidney disease 45
Chapter 3、 Materials and Methods 46
1. Animals selection and study design 46
2. Sample collection and processing 46
3. Analytes 47
4. Statistical Analysis 49
Chapter 4、 Results 51
1. Population 51
2. The comparative results 51
3. Effect of sex and age 51
4. Differences between healthy and KD group 51
5. The ability of blood CRE, UPC ratio, USG and uGGT index to detect abnormal renal ultrasonography in rabbits 52
Chapter 5、 Discussion 53
1. The comparative results of the urinary indices in healthy rabbits 53
2. The clinicopathological changes in KD rabbits 54
3. The use of blood CRE, UPC ratio, USG, uGGT index to detect abnormal renal condition/ ultrasonography in rabbits 58
4. Preliminary study of urine proteome by capillary gel electrophoretic analysis 59
5. Conclusion and limitations 59
Tables 61
Figures 69
References 75

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