Department of Comparative, Diagnostic, and Population Medicine, College of Veterinary Medicine, University of Florida, Gainesville, FL, 32610, USA.
Brevard Zoo and Sea Turtle Healing Center, Melbourne, FL, 32940, USA.
Vet Microbiol. 2021 Aug;259:109136. doi: 10.1016/j.vetmic.2021.109136. Epub 2021 Jun 18.
The genus Helicobacter includes spiral-shaped bacteria in the phylum Proteobacteria, class Epsilonproteobacteria, order Campylobacteriales, that have been associated with disease in animals, including reptiles. Three wild gopher tortoise (Gopherus polyphemus) index cases presented between 2012 and 2019 with nasal discharge, lethargy, and weight loss. Cytological examination of nasal discharge from all 3 tortoises identified marked heterophilic and mild histiocytic rhinitis with abundant extracellular and phagocytized spiral shaped bacteria that stained positive with Warthin-Starry stain. Polymerase chain reaction (PCR) and sequencing of the 16S rRNA gene revealed this to be a novel Helicobacter species. Two tortoises died despite treatment attempts, and the third was moribund and was euthanized. Histological examination of the nasal mucosa (n = 3) showed granulocytic to lymphocytic rhinitis with variable mucosal hyperplasia, erosion, and ulceration; Warthin-Starry staining highlighted the presence of spiral bacteria in the untreated tortoise. Genus-specific primers were designed, and the gyrA and groEL genes were amplified by PCR and sequenced. Phylogenetic analysis shows that this organism and other previously characterized Helicobacter from tortoises form a clade. Development and cross-validation of two qPCR diagnostic assays for the gyrA and groEL genes showed significant correlation of the results of two assays (P < 0.0001). These assays were used to survey nasal wash samples from 31 rehabilitating gopher tortoises. Mortality of tortoises significantly correlated with higher Helicobacter loads detected by qPCR (P = 0.028). Appropriate quarantine protocols for tortoises during rehabilitation should consider this organism. Upper respiratory disease in tortoises may involve complex microbial ecology; factors beyond Mycoplasmopsis (Mycoplasma) agassizii should be taken into account.
螺旋杆菌属包括变形菌门、γ-变形菌纲、弯曲杆菌目、弯曲杆菌科的螺旋形细菌，这些细菌与动物疾病有关，包括爬行动物。2012 年至 2019 年间，有 3 只野生地鼠龟（Gopherus polyphemus）索引病例出现鼻腔分泌物、嗜睡和体重减轻的症状。对这 3 只龟的鼻腔分泌物进行细胞学检查，发现明显的嗜中性粒细胞和轻度组织细胞性鼻炎，伴有丰富的细胞外和吞噬的螺旋形细菌，用华氏染色法染色呈阳性。聚合酶链反应（PCR）和 16S rRNA 基因测序显示这是一种新的螺旋杆菌属物种。尽管进行了治疗尝试，但其中 2 只龟死亡，而第 3 只龟病危并被安乐死。对 3 只龟的鼻黏膜（n = 3）进行组织学检查，发现粒细胞性到淋巴细胞性鼻炎，伴有不同程度的黏膜增生、糜烂和溃疡；华氏染色法突出显示未治疗的龟中存在螺旋形细菌。设计了属特异性引物，并通过 PCR 扩增和测序扩增 gyrA 和 groEL 基因。系统发育分析显示，该生物与其他以前从龟中分离出的螺旋杆菌属形成一个分支。gyrA 和 groEL 基因的 2 种 qPCR 诊断检测方法的开发和交叉验证显示，2 种检测方法的结果具有显著相关性（P < 0.0001）。使用这些检测方法对 31 只接受康复治疗的地鼠龟的鼻腔冲洗样本进行了检测。qPCR 检测到的螺旋杆菌属负荷较高与龟的死亡率显著相关（P = 0.028）。在康复期间，应考虑对龟进行适当的检疫隔离措施。龟的上呼吸道疾病可能涉及复杂的微生物生态系统；除了滑液支原体（Mycoplasma）agassizii 之外，还应考虑其他因素。
