Clinical pathology of amphibians: a review

Amphibian declines and extinctions have worsened in the last 2 decades. Partly because one of the main causes of the declines is infectious disease, veterinary professionals have increasingly become involved in amphibian research, captive husbandry, and management. Health evaluation of amphibians, free‐living or captive, can benefit from employing the tools of clinical pathology, something that is commonly used in veterinary medicine of other vertebrates. The present review compiles what is known of amphibian clinical pathology emphasizing knowledge that may assist with the interpretation of laboratory results, provides diagnostic recommendations for common amphibian diseases, and includes RIs for a few amphibian species estimated based on peer‐reviewed studies. We hope to encourage the incorporation of clinical pathology in amphibian practice and research, and to highlight the importance of applying veterinary medicine principles in furthering our knowledge of amphibian pathophysiology.     

Amphibian skin diseases.

Skin diseases in amphibian species seem to carry an additional degree of seriousness compared with those of other vertebrates. Because of the skin's importance in respiration and ion transport, breaching of the integument of these animals can result in fatal septicemias or metabolic disturbances. The timely diagnosis and treatment of these skin lesions is important. A review of the diseases affecting amphibian skin is provided. This article also describes the clinical signs, appropriate diagnostic steps, and treatment and control of amphibian skin diseases.     

Histological evidence of chytridiomycete fungal infection in a free-ranging amphibian,Afrana fuscigula (Anura: Ranidae), in South Africa

The 1st recorded histological evidence of chytridiomycete fungal infection in a free-ranging ranid amphibian in South Africa is presented. Literature on causes of a worldwide decline in amphibian populations is briefly reviewed.     

Advances in Amphibian Clinical Therapeutics

This review covers recent developments in the field of amphibian therapeutics without repeating previous extensive clinical reviews, formularies, and compendia. The information provided in this article would aid the veterinary practitioner treating amphibian species through the use of updated clinical therapeutic techniques and dosages.     

Cytologic diagnosis of disease in amphibians.

Cytology is an inexpensive yet powerful diagnostic tool that allows for rapid diagnosis of many common disease conditions in amphibian patients. Although the emphasis of this article is on infectious diseases, there is great potential for application of cytologic diagnosis to variety of medical conditions as the knowledge base in amphibian medicine and pathology continues to grow. Routine methods used that may fall under the umbrella of cytology range from wet mount examination of skin scrapings (or gill biopsies of larvae) to examination of stained impression smears. Routine Romanowsky's-type stains work well for amphibian samples. Preparation of multiple smears is always recommended to allow for use of special staining procedures.     

Amphibian virology.

Amphibians are a diverse group of species; much work remains to be done to elucidate the viruses of amphibians. Viral diseases may play an important role in wild and captive amphibian populations. Iridoviruses are a leading cause of wild amphibian mortality events in the United States and are a common cause of amphibian mortality events worldwide. In addition to frank viral diseases, viral infections may play a role in the establishment of bacterial, fungal, and parasitic diseases and are an underlying cause of neoplasia. It is important for the amphibian clinician to recognize disease syndromes and pathology that are consistent with viral etiology.     

The amphibian eye.

Knowledge of the anatomy of the amphibian eye is important to differentiate normal from abnormal. Special features of the amphibian eye mark the transition from aquatic to terrestrial life. The most often reported spontaneous ocular disease in the anurans (frogs and toads) is lipid keratopathy. An experimental study showed that high dietary cholesterol played a significant role in the development of corneal lipid infiltration. Other diseases, like panophthalmitis, occur with bacterial septicemia.     

Toll样受体信号通路在两栖类中的研究进展

两栖类处于脊椎动物由水生向陆生进化的过渡阶段,进化地位重要。近年来因疾病的爆发、生境破碎化、环境污染、紫外辐射增加、人为过度捕捉和生物入侵等诸多因素引起的全球范围内两栖类种群数量锐减已引起人们的广泛关注。Toll样受体(Toll-like receptor,TLR)家族是一类从线虫到哺乳动物普遍存在的高度保守的模式识别受体(PRRs),可以识别侵入机体病原体的病原相关分子模式(PAMPs),是两栖类先天性免疫防御系统的重要组成部分。本文结合近些年国内外对两栖类TLRs的研究对两栖类TLRs结构特征、进化特点、在两栖类发育早期的差异表达特征和两栖类TLRs信号通路中相关分子的表达研究进行概述。目前对于两栖类TLRs的研究发现两栖类TLRs兼具鱼类和哺乳类TLRs的特征,并在从鱼类到两栖类到哺乳类的进化过程中,TLRs家族部分成员如TLR2的配体识别功能可能发生了一定程度的特化;在两栖类早期发育阶段,获得性免疫不够完善,以TLRs为主的先天免疫防御系统在抵御病原体的侵袭方面发挥了重要作用;在特定病原菌和病毒的胁迫下,可能激活了TLRs下游不同的途径参与两栖类对病原体的免疫应答反应。     

Basic husbandry and clinical assessment of the amphibian patient.

The veterinarian presented with an amphibian patient must be prepared to assess both the animal's medical condition and its husbandry record; good health is inextricably linked to proper care and diet. This article provides the clinician with guidelines for maintaining amphibians in captivity, including information on climate control and lighting, housing and cage enrichment, and nutrition. The article also covers questions to ask when taking a history, methods of restraint, and practical advice on the equipment and techniques used to conduct a complete physical examination of the amphibian patient.     

福州市野生动物救护情况的统计分析

福州动物园于2009-2013年共救护野生动物70种1 033只(羽、头、尾),其中福建省有分布的动物种类占70%。在救护种类上鸟类最多有32种,其次为爬行类、兽类、两栖类;而在救护数量上爬行类最多,有633尾,其次为鸟类、两栖类、兽类。救护动物的主要来源为林业执法部门罚没。通过对福州动物园5年来救护动物的种类、数量、保护级别、动物分布情况、救护病因等情况进行总结分析,为今后开展野生动物救护提供一定的思路和对策。     

Chytridiomycosis in amphibians--first report in Europe

Declining of amphibian populations is a worldwide phenomenon. A cutaneous mycosis as a cause of death in free-living amphibians as well as in captive ones due to an chytrid fungus (Batrachochytrium dendrobatidis) was reported at first in 1998. This infections were reported hitherto from Australia, North, Central and South America. This is the first report on chytrid infections in captive anurans from Europe. Dendrobates auratus and D. pumilo imported from Costa Rica and P. vittatus imported from French Guayana died with chytridiomycosis within a week after arrival in Europe. Batrachocytrium was also found on captive bred frogs in Germany and Belgium. Clinical signs, diagnosis and conclusions for protecting free-living amphibian populations and captive frogs are discussed.     

Phylogenic aspects of the amphibian dual olfactory system

The phylogenic significance of the subdivision of dual olfactory system is reviewed mainly on the basis of our findings by electron microscopy and lectin histochemistry in the three amphibian species. The dual olfactory system is present in common in these species and consists of the projection from the olfactory epithelium (OE) to the main olfactory bulb (MOB) and that from the vomeronasal epithelium (VNE) to the accessory olfactory bulb (AOB). The phylogenic significance of subdivisions in the dual olfactory system in the amphibian must differently be interpreted. The subdivision of the MOB into its dorsal region (D-MOB) and ventral region (V-MOB) in Xenopus laevis must be attributed to the primitive features in their olfactory receptors. The middle cavity epithelium lining the middle cavity of this frog possesses both ciliated sensory cells and microvillous sensory cells, reminding the OE in fish. The subdivision of the AOB into the rostral (R-AOB) and caudal part (C-AOB) in Bufo japonicus formosus must be regarded as an advanced characteristic. The lack of subdivisions in both MOB and AOB in Cynops pyrrhogaster may reflect their phylogenic primitiveness. Since our lectin histochemistry to detect glycoconjugates expressed in the olfactory pathway reveals the subdivisions in the dual olfactory system in the amphibian, the glycoconjugates may deeply participate in the organization and function of olfactory pathways in phylogeny.     

两栖类结冰耐受机制及其研究策略

综述了近年来美洲林蛙的研究进展,总结了两栖类动物结冰耐受的主要机制,并提出该领域研究的新方向。     

Amphibian emergency medicine.

General concepts of amphibian emergency medicine are presented, including a review of patient selection, appropriate history, and helpful equipment. Physical examination procedures, general treatment, and diagnostic techniques are discussed. This is followed by a review of conditions commonly observed in amphibians and specific treatment options.     

Amphibian therapeutics.

The treatment of diseases in amphibians presents unique challenges for the clinician. This article describes the basic principles in selecting and administering therapeutics, and provides drug dose rates used successfully in amphibians. The importance of proper husbandary and hygiene, as it relates to amphibian health and medical management of disease, is also emphasized.     

Oral biology and conditions of amphibians.

This article reviews the anatomy of the skull and oral cavity in the three amphibian orders. Feeding behavior and feeding physiology are also described. A review is given of reported oral pathology. The pathology is divided into infectious causes (bacterial and parasitic) and noninfectious causes (trauma, neoplasia, and nutritional).     

Combining spawn egg counts,individual photo-ID and genetic fingerprinting to estimate the population size and sex ratio of an endangered amphibian

According to the International Union for Conservation of Nature Red List, 41% of the world's amphibian species are threatened with extinction, making them more threatened than any other vertebrate group nowadays. Given the global amphibian crisis, comprehensive understanding of demographics and population trends of declining and threatened species is essential for effective management and conservation strategies. Counting egg spawns is widely used to assess population abundance in pond breeding anurans. However, it is unknown how such counts translate into robust population size estimations. We monitored the breeding activity of the Natterjack toad (Epidalea calamita), combining egg string counts and individual photo-identification with Capture-Mark-Recapture population size and operational sex ratio estimation. Male Natterjack toads were identified by the pattern of natural markings with repeated ID of the same individual confirmed for 10% of the samples using genetic fingerprinting. We identified 647 unique individuals within a closed study population at Caherdaniel, Co Kerry. Population estimates derived from egg string counts estimated a breeding population of 368 females (95% CI 353–384) and Capture-Mark-Recapture estimated a breeding population of 1698 males (95% CI 1000–2397). The female:male sex ratio was conservatively estimated at 1:5 (95% CI 1:3–1:6) where 62% ± 6% of females were assumed to spawn. These substantially departed from any priori assumption of 1:1 which could have underestimated the breeding population by up to 72%. Where amphibian absolute population size estimation is necessary, methods should include empirical survey data on operational sex ratios and not rely on assumptions or those derived from the literature which may be highly population and/or context-dependent.     

Genome architecture changes and major gene variations of Andrias davidianus ranavirus (ADRV)

Ranaviruses are emerging pathogens that have led to global impact and public concern. As a rarely endangered species and the largest amphibian in the world, the Chinese giant salamander, Andrias davidianus, has recently undergone outbreaks of epidemic diseases with high mortality. In this study, we isolated and identified a novel ranavirus from the Chinese giant salamanders that exhibited systemic hemorrhage and swelling syndrome with high death rate in China during May 2011 to August 2012. The isolate, designated Andrias davidianus ranavirus (ADRV), not only could induce cytopathic effects in different fish cell lines and yield high viral titers, but also caused severely hemorrhagic lesions and resulted in 100% mortality in experimental infections of salamanders. The complete genome of ADRV was sequenced and compared with other sequenced amphibian ranaviruses. Gene content and phylogenetic analyses revealed that ADRV should belong to an amphibian subgroup in genus Ranavirus, and is more closely related to frog ranaviruses than to other salamander ranaviruses. Homologous gene comparisons show that ADRV contains 99%, 97%, 94%, 93% and 85% homologues in RGV, FV3, CMTV, TFV and ATV genomes respectively. In addition, several variable major genes, such as duplicate US22 family-like genes, viral eukaryotic translation initiation factor 2 alpha gene and novel 75L gene with both motifs of nuclear localization signal (NLS) and nuclear export signal (NES), were predicted to contribute to pathogen virulence and host susceptibility. These findings confirm the etiologic role of ADRV in epidemic diseases of Chinese giant salamanders, and broaden our understanding of evolutionary emergence of ranaviruses.     

A squamous cell carcinoma and an adenocarcinoma in Australian treefrogs

We describe the pathology in a captive adult White lipped treefrog (Litoria infrafrenata) with a squamous cell carcinoma of the skin and a free-living adult Common green treefrog (L. caerulea) with an adenocarcinoma of the skin. Although many amphibian neoplasms have been reported world wide, none had been described in Australian species.     

江西凌云山自然保护区两栖爬行动物多样性调查与区系分析

为掌握江西宁都凌云山省级自然保护区野生动物资源现状,促进自然保护区的有效管理和建设,作者于2012年至2013年多次对该保护区两栖爬行动物资源进行了实地考察。调查结果显示,凌云山保护区现已记录两栖爬行动物52种,隶属于2目13科37属。其中,两栖动物19种,隶属于1目6科14属,占江西省39种两栖动物的48. 7%;爬行动物33种,隶属于1目7科23属,占江西省80种的41. 3%。凌云山位于动物地理分布区中的东洋区系区的偏北位置,向古北区靠近,古北区的种类也会扩散到这一地区,成为东洋区、古北区的共有种。因此在两栖爬行种类组成上以东洋区的种类为主,具有多样性较为丰富的特点。最后对凌云山保护区的保护与管理工作提出了加大对当地群众的宣传和教育工作等建议。