Molecular characterization of a reptarenavirus detected in a Colombian Red-Tailed Boa (Boa constrictor imperator).

The global decline in biodiversity is a matter of great concern for members of the class Reptilia. Reptarenaviruses infect snakes, and have been linked to various clinical conditions, such as Boid Inclusion Body Disease (BIBD) in snakes belonging to the families Boidae and Pythonidae. However, there is a scarcity of information regarding reptarenaviruses found in snakes in both the United States and globally. This study aimed to contribute to the understanding of reptarenavirus diversity by molecularly characterizing a reptarenavirus detected in a Colombian Red-Tailed Boa (Boa constrictor imperator). Using a metagenomics approach, we successfully identified, and de novo assembled the whole genomic sequences of a reptarenavirus in a Colombian Red-Tailed Boa manifesting clinically relevant symptoms consistent with BIBD. The analysis showed that the Colombian Red-Tailed Boa in this study carried the University of Giessen virus (UGV-1) S or S6 (UGV/S6) segment and L genotype 7. The prevalence of the UGV/S6 genotype, in line with prior research findings, implies that this genotype may possess specific advantageous characteristics or adaptations that give it a competitive edge over other genotypes in the host population. This research underscores the importance of monitoring and characterizing viral pathogens in captive and wild snake populations. Knowledge of such viruses is crucial for the development of effective diagnostic methods, potential intervention strategies, and the conservation of vulnerable reptilian species. Additionally, our study provides valuable insights for future studies focusing on the evolutionary history, molecular epidemiology, and biological properties of reptarenaviruses in boas and other snake species.

Boid Inclusion Body Disease Is Also a Disease of Wild Boa Constrictors.

Reptarenaviruses cause boid inclusion body disease (BIBD), a potentially fatal disease, occurring in captive constrictor snakes boas and pythons worldwide. Classical BIBD, characterized by the formation of pathognomonic cytoplasmic inclusion bodies (IBs), occurs mainly in boas, whereas in pythons, for example, reptarenavirus infection most often manifests as central nervous system signs with limited IB formation. The natural hosts of reptarenaviruses are unknown, although free-ranging/wild constrictor snakes are among the suspects. Here, we report BIBD with reptarenavirus infection in indigenous captive and wild boid snakes in Costa Rica using histology, immunohistology, transmission electron microscopy, and next-generation sequencing (NGS). The snakes studied represented diagnostic postmortem cases of captive and wild-caught snakes since 1989. The results from NGS on archival paraffin blocks confirm that reptarenaviruses were already present in wild boa constrictors in Costa Rica in the 1980s. Continuous sequences that were de novo assembled from the low-quality RNA obtained from paraffin-embedded tissue allowed the identification of a distinct pair of reptarenavirus S and L segments in all studied animals; in most cases, reference assembly could recover almost complete segments. Sampling of three prospective cases in 2018 allowed an examination of fresh blood or tissues and resulted in the identification of additional reptarenavirus segments and hartmanivirus coinfection. Our results show that BIBD is not only a disease of captive snakes but also occurs in indigenous wild constrictor snakes in Costa Rica, suggesting boa constrictors to play a role in natural reptarenavirus circulation. IMPORTANCE The literature describes cases of boid inclusion body disease (BIBD) in captive snakes since the 1970s, and in the 2010s, others and ourselves identified reptarenaviruses as the causative agent. BIBD affects captive snakes globally, but the origin and the natural host of reptarenaviruses remain unknown. In this report, we show BIBD and reptarenavirus infections in two native Costa Rican constrictor snake species, and by studying archival samples, we show that both the viruses and the disease have been present in free-ranging/wild snakes in Costa Rica at least since the 1980s. The diagnosis of BIBD in wild boa constrictors suggests that this species plays a role in the circulation of reptarenaviruses. Additional sample collection and analysis would help to clarify this role further and the possibility of, e.g., vector transmission from an arthropod host.

Arsenicum albumas a treatment for pneumonia in snakes: three serial cases

two Python bivittatussnakes were received at Project Selva Viva, a zoo in Taubaté, Brazil, both presenting respiratory noises and oro-nasal discharge. A thoracic ultrasonographic examination was performed for evaluation on February 25th,2022, which diagnosed the presence of multiple vertical hyperechoic artifactsemerging from the pleural line, coalescing in some of the examined areas, and the presence a hypoechoic structure located in the subpleural region. Another serpent, from the Boa constrictor species, was apprehended and arrived at the same zoo without medical history, showing a low body score. During a thoracic ultrasonographic evaluation realized on April 1st,2022, B lines emerging from the pleural line were found. These vertical reverberating lines are a result of respiratory illness (SOLDATI et al., 2014).Methodology:The report was authorized by the owner of the zoo. All the patients received homeopathic therapy with 2 globules of Arsenicum album30 cH/ BID into the mouth, after being diagnosed with the respiratory condition by the ultrasonographic examination. The medication was chosen according to the similarity with the symptoms. The snakes had a runny nose and hissing breathing noise. Weekly ultrasound scans were performed on the Python bivittatus snakes to follow up on the respiratory condition. Results: On March 04th, it was observed that both snakes showed a reduction in respiratory noises and were more active. Ars 30 cH was maintained BID. On March 25th, both presented significant improvement in the ultrasound images, which showed only A lines, compatible with a healthy lung, and the treatment was suspended. Boa constrictor snake ́streatment started on April 1st. On April 8th, the ultrasonographic examination performed only presented A lines, which are characterized in ultrasound by parallel horizontal lines in the near field with the loss of image continuity in the far field, indicating improvement of the condition (LICHTENSTEIN et al., 2003). Conclusion:Given these results, the homeopathic treatment is an option to be considered for the treatment of respiratory symptoms in snakes, although the duration of the therapy varies based on the stage and chronicity of the disease.

Genetic characterisation of a novel reptarenavirus detected in a dead pet red-tailed boa (Boa constrictor).

Boid inclusion body disease (BIBD) is a severe and transmissible disease of snakes worldwide. Reptarenaviruses have been identified as the aetiological agents of BIBD. We determined the almost complete genome sequence of an arenavirus detected in a female red-tailed boa that had succumbed in a private collection in Hungary. We used a combination of next generation sequencing and Sanger sequencing methods. Based on the analysis of the obtained sequence data, the virus, tentatively named Coldvalley virus, seemed to belong to the Reptarenavirus genus of the Arenaviridae family. This classification was confirmed by the genome structure (bisegmented single-stranded RNA) characteristic of the genera Mammarenavirus and Reptarenavirus. The pairwise comparison of the nucleotide and amino acid sequences, as well as the topology of the maximum likelihood phylogenetic trees, suggested that the newly-characterised Coldvalley virus can be classified into the species Rotterdam reptarenavirus.

Functional activation of the periaqueductal gray matter during conditioned and unconditioned fear in guinea pigs confronted with the Boa constrictor constrictor snake

The periaqueductal gray matter (PAG) is an essential structure involved in the elaboration of defensive responses, such as when facing predators and conspecific aggressors. Using a prey vs predator paradigm, we aimed to evaluate the PAG activation pattern evoked by unconditioned and conditioned fear situations. Adult male guinea pigs were confronted either by a Boa constrictor constrictor wild snake or by the aversive experimental context. After the behavioral test, the rodents were euthanized and the brain prepared for immunohistochemistry for Fos protein identification in different PAG columns. Although Fos-protein-labeled neurons were found in different PAG columns after both unconditioned and conditioned fear situations at the caudal level of the PAG, we found greater activation of the lateral column compared to the ventrolateral and dorsomedial columns after predator exposure. Moreover, the lateral column of the PAG showed higher Fos-labeled cells at the caudal level compared to the same area at the rostral level. The present results suggested that there are different activation patterns of PAG columns during unconditioned and conditioned fear in guinea pigs. It is possible to hypothesize that the recruitment of specific PAG columns depended on the nature of the threatening stimulus.

Monitoring of the Reproductive Cycle in Captive-Bred Female Boa constrictor: Preliminary Ultrasound Observations.

The Boa constrictor is one of the most common reptiles bred in captivity. To achieve a successful breeding season, thorough knowledge of the females' reproductive activity is necessary. In this regard, information on the Boa constrictor is still rather scarce. The aim of the present study was to monitor the ovarian activity and the embryonic development of boas by ultrasound. We performed brief scans on thirty non-anaesthetized snakes using a portable ultrasound system and a 7.5-10 MHz linear array transducer (Esaote MyLab™ Classic). Ultrasound features, dimensions, and echogenicity of the preovulatory and postovulatory follicles were determined. As gestation progresses, the postovulatory follicle size increases, and the embryonic silhouette becomes increasingly recognizable. During the second month after ovulation, by using color Doppler, early embryos' heart activity could be evaluated. It is possible to highlight vascular connections between the mother and the membrane covering the embryonic structures. Ultrasound also allows one to identify follicular regression or slugs (nonfertilized eggs) early. The present study suggests that ultrasound could be an excellent noninvasive technique to evaluate the reproductive activity of Boa constrictor, allowing us to precisely identify the correct time for mating, monitor embryo development and viability, and allow the early diagnosis of follicular regression.

Ultrasonographic characterization of the organs in the middle third and caudal coelomic cavity of snakes in the Boidae Family

ABSTRACT: The aim of this study was the ultrasonographic evaluation of the organs in the middle third and caudal coelomic cavity of healthy snakes in the Boidae family. For such, 15 adult snakes from five species were evaluated: Boa constrictor amarali, Boa constrictor constrictor, Eunectes murinus, Epicrates assisi and Epicrates crassus. The animals were physically restrained for examination. After the application of acoustic gel over the scales the transducer was positioned in the frontal plane of the coelomic cavity. Microconvex and linear transducers at 10MHz were used. The ultrasound examination was conducted in the craniocaudal direction, with longitudinal and transversal sections to assess syntropy, echogenicity and echotexture of the organs. The liver was bilobulated, hyperechoic, homogeneous echotexture, hyperechoic capsule, with a hepatic lobe measuring 1.15±0.64cm wide, and the presence of a central hepatic vein, measuring 0.44±0.21cm in diameter. The gallbladder was characterized as an anechoic structure with echogenic walls and dimensions of 2.91±1.18cm x 1.38±0.84cm (length x width). The stomach showed a pleated and linear texture, echogenic, with hypoechoic walls measuring 0.3±0.07cm wide. The splenopancreas was a circular and echogenic structure, homogeneous echotexture and measuring 1.18±0.62cm in width and 1.56±0.88cm in length. The kidneys were characterized by elongated, lobulated, hypoechoic structures, an echogenic capsule and an echogenic central line, measuring 1.05±0.7cm in width. The testis were characterized by fusiform, hypoechoic structures, homogeneous echotexture, measuring 2.31±0.79cm in length and 0.6±0.23cm in width. Ovarian follicles were seen in different stages, 0.67±0.39cm wide and 0.73±0.38cm long. No differences were found between species of syntropy, echogenicity and echotexture of the organs of the coelomic cavity. The ultrasound examination proved to be a safe, non-invasive and efficient technique for characterizing the organs of the coelomic cavity of snakes.

RESUMO: O objetivo do estudo foi realizar a avaliação ultrassonográfica de órgãos do terço médio e caudal da cavidade celomática de serpentes hígidas da família Boidae. Para isto, foram avaliadas 15 serpentes adultas de cinco espécies: Boa constrictor amarali, Boa constrictor constrictor, Eunectes murinus, Epicrates assisi e Epicrates crassus. Os animais foram contidos fisicamente para a realização do exame. Após aplicação de gel acústico sobre as escamas, o transdutor foi posicionado no plano frontal da cavidade celomática. Foram utilizados transdutores microconvexo e linear em 10MHz. O exame ultrassonográfico foi conduzido no sentido craniocaudal, com cortes longitudinais e transversais para avaliar sintopia, ecogenicidade e ecotextura dos órgãos. O fígado se apresentou bilobulado, hiperecoico, ecotextura homogênea, cápsula hiperecoica, com lobo hepático medindo 1,15±0,64cm de largura, e presença de uma veia central hepática, medindo 0,44±0,21cm de diâmetro. A vesícula biliar foi caracterizada como uma estrutura anecoica de paredes ecogênicas e dimensões de 2,91±1,18cm x 1,38±0,84cm(comprimento x largura). O estômago apresentou-se com o aspecto pregueado e linear, ecogênico, com paredes hipoecoicas medindo 0,3±0,07cm de largura. O esplenopâncreasse apresentou como uma estrutura circular e ecogênica, ecotextura homogênea e medindo de 1,18±0,62cm de largura e 1,56±0,88cm de comprimento. Os rins se caracterizaram por estruturas alongadas, lobuladas, hipoecoicas, cápsula ecogênica e uma linha central ecogênica, medindo 1,05±0,7cm de largura. Os testículos foram caracterizados por estruturas fusiformes, hipoecoicas, ecotextura homogênea, medindo 2,31±0,79cm de comprimento e 0,6±0,23cm de largura. Foram visualizados folículos ovarianos em diferentes estágios, de 0,67±0,39cm de largura e 0,73±0,38cm de comprimento. Não foram encontradas diferenças entre espécies de sintopia, ecogenicidade e ecotextura dos órgãos da cavidade celomática. O exame ultrassonográfico demonstrou-se uma técnica segura, não invasiva e eficiente para caracterização dos órgãos da cavidade celomática de serpentes.

Ultrasonographic characterization of the organs in the middle third and caudal coelomic cavity of snakes in the Boidae Family / Caracterização ultrassonográfica dos órgãos do terço médio e caudal da cavidade celomática de serpentes da família Boidae

The aim of this study was the ultrasonographic evaluation of the organs in the middle third and caudal coelomic cavity of healthy snakes in the Boidae family. For such, 15 adult snakes from five species were evaluated: Boa constrictor amarali, Boa constrictor constrictor, Eunectes murinus, Epicrates assisi and Epicrates crassus. The animals were physically restrained for examination. After the application of acoustic gel over the scales the transducer was positioned in the frontal plane of the coelomic cavity. Microconvex and linear transducers at 10MHz were used. The ultrasound examination was conducted in the craniocaudal direction, with longitudinal and transversal sections to assess syntropy, echogenicity and echotexture of the organs. The liver was bilobulated, hyperechoic, homogeneous echotexture, hyperechoic capsule, with a hepatic lobe measuring 1.15±0.64cm wide, and the presence of a central hepatic vein, measuring 0.44±0.21cm in diameter. The gallbladder was characterized as an anechoic structure with echogenic walls and dimensions of 2.91±1.18cm x 1.38±0.84cm (length x width). The stomach showed a pleated and linear texture, echogenic, with hypoechoic walls measuring 0.3±0.07cm wide. The splenopancreas was a circular and echogenic structure, homogeneous echotexture and measuring 1.18±0.62cm in width and 1.56±0.88cm in length. The kidneys were characterized by elongated, lobulated, hypoechoic structures, an echogenic capsule and an echogenic central line, measuring 1.05±0.7cm in width. The testis were characterized by fusiform, hypoechoic structures, homogeneous echotexture, measuring 2.31±0.79cm in length and 0.6±0.23cm in width. Ovarian follicles were seen in different stages, 0.67±0.39cm wide and 0.73±0.38cm long. No differences were found between species of syntropy, echogenicity and echotexture of the organs of the coelomic cavity. The ultrasound examination proved to be a safe, non-invasive and efficient technique for characterizing the organs of the coelomic cavity of snakes.(AU)

O objetivo do estudo foi realizar a avaliação ultrassonográfica de órgãos do terço médio e caudal da cavidade celomática de serpentes hígidas da família Boidae. Para isto, foram avaliadas 15 serpentes adultas de cinco espécies: Boa constrictor amarali, Boa constrictor constrictor, Eunectes murinus, Epicrates assisi e Epicrates crassus. Os animais foram contidos fisicamente para a realização do exame. Após aplicação de gel acústico sobre as escamas, o transdutor foi posicionado no plano frontal da cavidade celomática. Foram utilizados transdutores microconvexo e linear em 10MHz. O exame ultrassonográfico foi conduzido no sentido craniocaudal, com cortes longitudinais e transversais para avaliar sintopia, ecogenicidade e ecotextura dos órgãos. O fígado se apresentou bilobulado, hiperecoico, ecotextura homogênea, cápsula hiperecoica, com lobo hepático medindo 1,15±0,64cm de largura, e presença de uma veia central hepática, medindo 0,44±0,21cm de diâmetro. A vesícula biliar foi caracterizada como uma estrutura anecoica de paredes ecogênicas e dimensões de 2,91±1,18cm x 1,38±0,84cm(comprimento x largura). O estômago apresentou-se com o aspecto pregueado e linear, ecogênico, com paredes hipoecoicas medindo 0,3±0,07cm de largura. O esplenopâncreasse apresentou como uma estrutura circular e ecogênica, ecotextura homogênea e medindo de 1,18±0,62cm de largura e 1,56±0,88cm de comprimento. Os rins se caracterizaram por estruturas alongadas, lobuladas, hipoecoicas, cápsula ecogênica e uma linha central ecogênica, medindo 1,05±0,7cm de largura. Os testículos foram caracterizados por estruturas fusiformes, hipoecoicas, ecotextura homogênea, medindo 2,31±0,79cm de comprimento e 0,6±0,23cm de largura. Foram visualizados folículos ovarianos em diferentes estágios, de 0,67±0,39cm de largura e 0,73±0,38cm de comprimento. Não foram encontradas diferenças entre espécies de sintopia, ecogenicidade e ecotextura dos órgãos da cavidade celomática. O exame ultrassonográfico demonstrou-se uma técnica segura, não invasiva e eficiente para caracterização dos órgãos da cavidade celomática de serpentes.(AU)

Virtopsy of a gravid Boa constrictor using computed tomography and magnetic resonance imaging.

This article presents radiologic examinations of a deeply sedated Boa constrictor with boid inclusion body disease (BIBD) as an adjunction to the subsequent necropsy. This method is known as virtopsy. The Boa constrictor in the present case was gravid. Computed tomography (CT) allowed for the detailed depiction of a fetal skeleton at the rear end of the adult snake. Furthermore, tiny gas formation was detected inside the cranium of the fetus, which was deemed a radiologic sign for decomposition. Magnetic resonance imaging (MRI) delineated the soft tissue at high resolution. This article illustrates the use of CT and MRI for the examination of a gravid Boa constrictor before necropsy and demonstrates the detection of "normal" postmortem findings leading to the confirmation of fetal death in situ.

Identification of Reptarenaviruses, Hartmaniviruses, and a Novel Chuvirus in Captive Native Brazilian Boa Constrictors with Boid Inclusion Body Disease.

Boid inclusion body disease (BIBD) is a transmissible viral disease of captive snakes that causes severe losses in snake collections worldwide. It is caused by reptarenavirus infection, which can persist over several years without overt signs but is generally associated with the eventual death of the affected snakes. Thus far, reports have confirmed the existence of reptarenaviruses in captive snakes in North America, Europe, Asia, and Australia, but there is no evidence that it also occurs in wild snakes. BIBD affects boa species within the subfamily Boinae and pythons in the family Pythonidae, the habitats of which do not naturally overlap. Here, we studied Brazilian captive snakes with BIBD using a metatranscriptomic approach, and we report the identification of novel reptarenaviruses, hartmaniviruses, and a new species in the family Chuviridae The reptarenavirus L segments identified are divergent enough to represent six novel species, while we found only a single novel reptarenavirus S segment. Until now, hartmaniviruses had been identified only in European captive boas with BIBD, and the present results increase the number of known hartmaniviruses from four to six. The newly identified chuvirus showed 38.4%, 40.9%, and 48.1% amino acid identity to the nucleoprotein, glycoprotein, and RNA-dependent RNA polymerase, respectively, of its closest relative, Guangdong red-banded snake chuvirus-like virus. Although we cannot rule out the possibility that the found viruses originated from imported snakes, the results suggest that the viruses could circulate in indigenous snake populations.IMPORTANCE Boid inclusion body disease (BIBD), caused by reptarenavirus infection, affects captive snake populations worldwide, but the reservoir hosts of reptarenaviruses remain unknown. Here, we report the identification of novel reptarenaviruses, hartmaniviruses, and a chuvirus in captive Brazilian boas with BIBD. Three of the four snakes studied showed coinfection with all three viruses, and one of the snakes harbored three novel reptarenavirus L segments and one novel S segment. The samples originated from collections with Brazilian indigenous snakes only, which could indicate that these viruses circulate in wild snakes. The findings could further indicate that boid snakes are the natural reservoir of reptarena- and hartmaniviruses commonly found in captive snakes. The snakes infected with the novel chuvirus all suffered from BIBD; it is therefore not possible to comment on its potential pathogenicity and contribution to the observed changes in the present case material.

Effects of brodifacoum and diphacinone exposure on four species of reptiles: tissue residue levels and survivorship.

BACKGROUND: Anticoagulant rodenticides are used worldwide to control pest rodent species. However, the risks posed to non-target reptiles have not been well characterized. In this study, 46 giant ameivas (Ameiva ameiva), 39 boa constrictors (Boa constrictor), 33 wood turtles (Rhinoclemmys pulcherrima), and 47 green iguanas (Iguana iguana) were orally dosed with one of two levels of either diphacinone or brodifacoum anticoagulant in propylene glycol solutions. Dosages were derived using daily food intake (DFI) equations, converting DFI to an equivalent anticoagulant bait amount and gavaging the solution volume needed to deliver the quantity of anticoagulant in that amount of bait. Animals were dosed on days 0 and 7 and monitored for a further 7 days for signs of anticoagulant intoxication and differences in behaviors and postures. At necropsy on day 14, animals were examined for thoracic and abdominal bleeding, and both tissue and organ samples were taken for histology. Liver and whole-body anticoagulant residues were assessed. RESULTS: No turtles or boas died due to anticoagulant exposure. However, anticoagulant intoxication was suspected in one iguana dosed with brodifacoum. A few treated ameivas died but exhibited no hemorrhaging. Liver residue levels were higher than whole-body remainder residue levels for all species. Unlike the other species, turtles had higher diphacinone residue levels than brodifacoum. CONCLUSION: Turtles and boas exhibited a relative insensitivity to diphacinone and brodifacoum, while the lizards appeared to be somewhat more sensitive to these compounds. This study provides data for future assessments of the risks to these species associated with anticoagulant use. Published 2019. This article is a U.S. Government work and is in the public domain in the USA.

Genomic Basis of Convergent Island Phenotypes in Boa Constrictors.

Convergent evolution is often documented in organisms inhabiting isolated environments with distinct ecological conditions and similar selective regimes. Several Central America islands harbor dwarf Boa populations that are characterized by distinct differences in growth, mass, and craniofacial morphology, which are linked to the shared arboreal and feast-famine ecology of these island populations. Using high-density RADseq data, we inferred three dwarf island populations with independent origins and demonstrate that selection, along with genetic drift, has produced both divergent and convergent molecular evolution across island populations. Leveraging whole-genome resequencing data for 20 individuals and a newly annotated Boa genome, we identify four genes with evidence of phenotypically relevant protein-coding variation that differentiate island and mainland populations. The known roles of these genes involved in body growth (PTPRS, DMGDH, and ARSB), circulating fat and cholesterol levels (MYLIP), and craniofacial development (DMGDH and ARSB) in mammals link patterns of molecular evolution with the unique phenotypes of these island forms. Our results provide an important genome-wide example for quantifying expectations of selection and convergence in closely related populations. We also find evidence at several genomic loci that selection may be a prominent force of evolutionary change-even for small island populations for which drift is predicted to dominate. Overall, while phenotypically convergent island populations show relatively few loci under strong selection, infrequent patterns of molecular convergence are still apparent and implicate genes with strong connections to convergent phenotypes.

Distribution and properties of cardiac and pulmonary ß-adrenergic receptors in corn snakes (Pantherophis guttatus) and Boa constrictor (Boa constrictor).

The aim of the present study was to characterize ß-adrenergic receptors in the snake heart and lung of corn and Boa constrictor snakes. The ß-adrenergic receptor binding sites were studied in purified heart and lung membranes using the specific ß-adrenergic receptor antagonist [125J]-iodocyanopindolol (ICYP) and subtypes using selective ß1-adrenergic receptor antagonist CGP-20712A and selective ß2-adrenergic receptor antagonist ICI-118.551. A saturable and specific ß-adrenergic receptor binding site was detected in cardiac membranes with maximal receptor density (Bmax) of 43.99 ±â€¯3.86 fmol/mg protein (corn snake) and 58.07 ±â€¯2.88 fmol/mg protein (Boa constrictor) as well as KD of 24.21 ±â€¯7.38 pM (corn snake) and 21.48 ±â€¯3.85 pM (Boa constrictor) and in lung membranes (Bmax fmol/mg protein: 55.95 ±â€¯16.28 (corn snake) and 107.00 ±â€¯14.21 (Boa constrictor); KD pM: 71.25 ±â€¯21.92 (corn snake) and 55.04 ±â€¯18.68 (Boa constrictor)). Competition-binding studies showed ß-adrenergic receptors with low affinities to the ß2-selective adrenergic receptor antagonist and high affinity binding to ß1-selective adrenergic receptor antagonist in both heart and lung tissues of both snake species, suggesting the presence of high population of the post-synaptic ß1-adrenergic receptor subtype. It seems that the presence of the predominant ß1-subtype also in lung tissues may indicate the importance of the vascular system in the snake lung.

Identification and mapping of real acupoints in the anatomical topography of Boa constrictor / Identificação e mapeamento de acupontos reais na topografia anatômica de Boa constrictor

Abstract Acupuncture is a therapy of the traditional Chinese medicine technique, which over the years has been widespread in the West. Defined as a needle-point treatment technique (acupoints), exactly preconfigured in the body to produce a specific physiological reaction to restore the balance between the function of conflicting states and homeostasis, alternating states of energy and thus maintain the ideal organ and body organization. Thus, the objective of this study was to map and describe the main points of acupuncture in the species Boa constrictor , and their indications to promote the balance of this species. The unprecedented result of the mapping was the discovery of specific acupoints with individual location indications without distribution in specific meridians and dispersedly distributed in the body.

Resumo A acupuntura é uma técnica terapêutica da Medicina Tradicional Chinesa, que ao longo dos anos tem sido muito difundida no Ocidente. Definida como uma técnica de tratamento com inserção de agulhas em pontos (acupontos) exatamente pré-estabelecidos sobre o organismo, para produzir uma reação fisiológica específica com o intuito de restabelecer o equilíbrio entre estados contraditórios de função e a homeostase, alternando os estados de energia e com isto mantendo a organização do órgão e do organismo. Com isso, o objetivo do presente trabalho foi mapear e descrever os principais pontos de acupuntura nas espécies Boa constrictore suas indicações para promover o equilíbrio dessa espécie. O resultado inédito do mapeamento foi à descoberta de pontos de acupuntura específicos com indicações de localização individuais sem distribuição em meridianos específicos e dispersamente distribuídos no corpo.

Ebola virus, but not Marburg virus, replicates efficiently and without required adaptation in snake cells.

Ebola virus (EBOV) disease is a viral hemorrhagic fever with a high case-fatality rate in humans. This disease is caused by four members of the filoviral genus Ebolavirus, including EBOV. The natural hosts reservoirs of ebolaviruses remain to be identified. Glycoprotein 2 of reptarenaviruses, known to infect only boa constrictors and pythons, is similar in sequence and structure to ebolaviral glycoprotein 2, suggesting that EBOV may be able to infect reptilian cells. Therefore, we serially passaged EBOV and a distantly related filovirus, Marburg virus (MARV), in boa constrictor JK cells and characterized viral infection/replication and mutational frequency by confocal imaging and sequencing. We observed that EBOV efficiently infected and replicated in JK cells, but MARV did not. In contrast to most cell lines, EBOV-infected JK cells did not result in an obvious cytopathic effect. Surprisingly, genomic characterization of serial-passaged EBOV in JK cells revealed that genomic adaptation was not required for infection. Deep sequencing coverage (>10,000×) demonstrated the existence of only a single nonsynonymous variant (EBOV glycoprotein precursor pre-GP T544I) of unknown significance within the viral population that exhibited a shift in frequency of at least 10 per cent over six serial passages. In summary, we present the first reptilian cell line that replicates a filovirus at high titers, and for the first time demonstrate a filovirus genus-specific restriction to MARV in a cell line. Our data suggest the possibility that there may be differences between the natural host spectra of ebolaviruses and marburgviruses.

INVESTIGATION OF THE TRACHEAL MUCOCILIARY CLEARANCE IN SNAKES WITH AND WITHOUT BOID INCLUSION BODY DISEASE AND LUNG PATHOLOGY.

Pneumonia is a common complication of boid inclusion body disease (BIBD) in snakes. The tracheal mucociliary apparatus of eight boas ( Boa constrictor) and two pythons ( Python regius, Morelia viridis) was examined to assess whether absent or reduced mucociliary clearance could be a predisposing factor. Nine of the examined snakes were positive for BIBD by detection of inclusion bodies and three had lung pathologies other than the formation of inclusion bodies. A considerable individual variation of ciliary beat frequency (CBF, 3.0 ± 0.75 Hz to 7.8 ± 1.27 Hz), transport speed (23.1 ± 12.56 µm/sec to 189.2 ± 41.17 µm/sec), and transport direction (-12.5° ± 11.43° to 36.1° ± 7.53°) was found. CBFs of the BIBD-affected snakes with or without lung pathologies were markedly lower than ranges published for birds or mammals, but the net transport speeds and directions lay well within. The present investigation does therefore not reveal any signs of an inadequate mucociliary clearance in BIBD-affected snakes.

Uso de cetamina pela via cólon-retal para contenção química de jiboias Boa constrictor Linnaeus, 1758 com teste de influência da droga sobre o sistema renal / Use of ketamine by colon-rectal route for chemical restraint in Boa constrictor Linnaeus, 1758 with test of influence of the drug on renal system

Por serem frequentes na clínica de animais silvestres, faz-se necessária a manipulação de jiboias e para isso, indispensável o conhecimento sobre as manobras precisas para o tratamento das possíveis afecções. Quando se opta pela contenção química, ou pela realização de procedimentos cirúrgicos, um dos fármacos utilizados na anestesia de serpentes é a cetamina. Viu-se a necessidade de buscar uma via alternativa, semelhante em eficácia às tradicionais para a contenção química, porém que minimizasse os riscos e efeitos adversos encontrados na sua execução. O presente trabalho sugere que a via retal seja esta alternativa, por isso, treze jiboias foram submetidas à administração de 70mg/kg de cloridrato de cetamina, com sonda uretral, através da cloaca até o cólon-reto. Foram avaliados, nos tempos 0, 10, 20, 30, 40, 50, 60, 70, 80, 90 e 120 minutos, a partir da administração do fármaco, os seguintes parâmetros: frequência cardíaca, relaxamento muscular e mobilidade, resistência à contenção ou manipulação e reação postural de endireitamento. Foi realizada coleta de 0,5mL de sangue por punção do seio venoso paravertebral cervical, antes da administração do fármaco, no dia seguinte e após nove dias. Foram dosadas as concentrações plasmáticas de cálcio, fósforo e ácido úrico de todos os exemplares a fim de verificar o perfil bioquímico renal e avaliar a influência do fármaco neste sistema. Não foram observadas alterações bioquímicas plasmáticas durante o período de avaliação. Foi possível promover a contenção química das jiboias Boa constrictor, utilizando cloridrato de cetamina pela via cólon-retal.(AU)

It is necessary to deal with Boa constrictor snakes because they are frequently treated in wild and exotic animal clinics and the knowledge about the required procedures in the treatment of the possible affections becomes imperative. When the choice for chemical restraint or sedation for surgical procedures is made, one of the drugs used in snakes is ketamine. We believed it was necessary to look for an alternative route of drug administration as effective as the regular ones, but with minimum risks and less adverse effects in its execution. Therefore thirteen snakes were submitted to the administration of 70mg/kg of ketamine hydrochloride, with an urethral tube, through the cloaca into the colon-rectum. After this, they were evaluated during the 0, 10, 20, 30, 40, 50, 60, 70, 80, 90 and 120 next minutes from the administration time using the following parameters: heart rate, muscle relaxation and mobility, handling or restraint resistance and righting reflex. Blood samples were collected from each snake by cervical paravertebral venous sinus punction, before the drug administration, on the next day and nine days after. Serum concentrations of calcium, phosphorus and uric acid were measured in order to check the renal biochemical profile and the possibility of influence of the drug on this system. It was possible to provoke chemical restraint in Boa constrictor snakes, with ketamine hydrochloride administered by the colon-rectal route.(AU)

Concentração de elementos traços em serpentes do litoral e da região serrana do Espírito Santo / Trace element concentration in snakes of coastal and serrana region of the State of Espírito Santo, Brazil

Os elementos químicos, em sua maioria, encontram-se em ciclos bioquímicos e geoquímicos fechados e em concentrações que não causam efeitos nocivos aos organismos. Contudo, ações antrópicas aceleradas promovem alterações ambientais, como o aumento no aporte de contaminantes. Com essas informações, o presente trabalho buscou caracterizar a presença aos elementos traços em duas espécies de serpentes comuns na região sudeste, Boa constrictor e Bothrops jararaca. Os exemplares de B. constrictor (n=18) foram provenientes de atropelamentos ocorridos no trecho da Rodovia ES-060 do Km 0 ao Km 67,5. Os exemplares de B. jararaca (n=18) foram capturados por fazendeiros na zona rural da região serrana do Espírito Santo. Foram analisados 1 grama de fragmento de rim, de espécimes necropsiados. Os rins coletados foram digeridos com mistura ácida (HNO3;HCl; 1:1) a 300oC, 40min e a quantificação de microelementos (Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb e Zn) foi realizada por espectrometria de emissão óptica com plasma indutivamente acoplado (ICP OES). Não foram observadas diferenças significativas entre machos e fêmeas, exceto para Cromo nas B. constrictor (p=0,03), com média de 1,6595 nas concentrações de Cr nas fêmeas e 0,2896 em machos. Contudo, quando foram comparadas as diferentes espécies de serpentes, diferentes concentrações foram encontradas, para todos os elementos determinados, com destaque para o Ferro com uma concentração de 106,2mg/g em B. constrictor e 120,3mg/g em B. jararaca, provavelmente por esses animais virem de áreas próximas à portos de minério de ferro. A concentração de Zinco em B. constrictor foi de 1261,8mg/g e em B. jararaca foi de 28,4mg/g. O presente estudo indica que as serpentes analisadas, que habitam as regiões da Grande Vitória e serrana do Espírito Santo apresentam elevadas concentrações dos microelementos Zn e Fe.(AU)

Chemical elements are mostly involved in biochemical and geochemical cycles in concentrations which are harmless for organisms. However, accelerated human actions promote environmental changes such as the increase in contaminant intake. With this information, this study aimed to characterize the presence of trace elements in two snake species common in Southeastern Brazil, Boa constrictor and Bothrops jararaca. Copies of B. constrictor (n=18) were from pedestrian accidents which occurred on Highway stretch ES-060 from Km 0 to Km 67.5. Copies of B. jararaca (n=18) were captured by farmers in a rural mountainous area of Espíirito Santo State, Brazilo. They were analyzed 1 kidney fragment grass, autopsied specimens. Harvested kidneys were digested with acid mixture (HNO3 e HCl 1:1) at 300°C, 40 min and quantification of microelementos (Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn) were determined by spectrometric optical emission with inductively coupled plasma (ICP OES). significant differences between males and females were observed, except for Chrome in B. constrictor (p=0.03), with an average of 1.6595 in Cr concentration of females and 0.2896 in males. However, when different species of snakes were compared, different concentrations were found for all elements, especially iron with a concentration of 106.2mg/g in B. constrictor and 120.3mg/g in B. jararaca; probably these animals came from areas near iron ore ports. The concentration of zinc in B. constrictor was 1261.8mg/g and in B. jararaca 28,4mg/g. This study indicates that the analyzed snakes, which inhabit the regions of Greater Victoria and the mountainous region of the State of Espírito Santo have high concentrations of the microelements Zn and Fe.(AU)

Doença do corpúsculo de inclusão e espondilite por Salmonella sp. em uma Boa constrictor constrictor / Inclusion body disease and spondilitis by Salmonella sp. in a Boa constrictor constrictor

Doença do corpúsculo de inclusão (IBD) é uma enfermidade caracterizada por corpúsculos intracitoplasmáticos em diversos tecidos, principalmente no sistema nervoso central, responsável pelos principais sinais clínicos atribuídos à doença que acomete Boas e Phytons de cativeiro; essa enfermidade tem sido uma preocupação mundial devido à alta morbidade e mortalidade. O diagnóstico é feito pela visualização dos corpúsculos causados por um Arenavírus modificado. Salmonella sp. pertence à microflora de animais de sangue frio e quente, e é um patógeno oportunista que pode causar quadros gastrointestinais ou septicêmicos. Em répteis a Salmonella sp. é a bactéria com maior frequência de citações em espondilites e osteomielites. Relata-se um caso de uma jiboia (Boa constrictor constrictor) que apresentava restrição de movimento e múltiplos granulomas dorsais nas vértebras; à radiografia evidenciaram-se regiões fraturadas. Após meses de tratamentos sem melhora clínica e o aparecimento de novas lesões o animal ficou prostrado, anoréxico, caquético e desenvolveu opistótono; optou-se pela eutanásia. À necropsia verificaram-se, nas vértebras, múltiplos focos dorsais com aumento de volume que variava de 1,7cm à 3,8cm. Ao corte as vértebras eram deformadas e exibiam conteúdo caseoso focal próximo ao canal medular, este foi coletado para microbiologia onde se identificou Salmonella sp. À microscopia as vértebras tinham um infiltrado inflamatório multifocal moderado de macrófagos e heterofilos. Algumas áreas possuíam grande quantidade de granulomas com calcificação central e inúmeras células gigantes; outros mostravam áreas de osteomalácia e fibrose. Em raros focos havia fratura do corpo vertebral e compressão da medula espinhal com leve infiltrado inflamatório invadindo o canal medular. No pulmão, principalmente no epitélio brônquico, por vezes até dentro de linfócitos do tecido linfoide bronco-associado, no intestino, fígado, vesícula biliar, nos rins e no encéfalo foram encontradas diversas estruturas eosinofílicas intracitoplasmáticas arredondadas que variavam de 1 a 10 µm. Essas estruturas acompanhavam ou não inflamações mononucleares. Os achados são compatíveis com IBD e espondilite por salmonelose. A IBD é uma enfermidade frequente em serpentes de cativeiro, de importância mundial, que provavelmente é subdiagnosticada no Brasil. Essa doença causa imunossupressão que favorece ao desenvolvimento de outras enfermidades, e é tipicamente associada a outras doenças como a espondilite encontrada no caso.(AU)

Inclusion Body Disease (IBD) is a disorder characterized by intracytoplasmic corpuscles in different tissues, mainly in the CNS, wich is responsible for the major neurological signs attributable to this disease. It affects Boas and Phytons in captivity and have been a global concern due to the high morbidity and mortality. The diagnosis is made by visualization of corpuscles caused by a modified Arenaviruses. Salmonella sp. belongs to microflora of cold and warm-blooded animals; it is an opportunistic pathogen that can causes gastrointestinal or septic disorders. In reptiles, Salmonella sp. is the bacteria most frequently quotes in spondylitis and osteomyelitis. This article describes a boa constrictor (Boa constrictor constrictor) that had restriction of movement and multiple granulomas in the dorsal vertebrae, the shadowgraph showed up fractured regions. After months of treatment without clinical improvement and the emergence of new injuries, the animal started to get prostrate, anorexic, cachectic and developed opisthotonos. It was opted for euthanasia. At necropsy it was found in multiple spots swelling of the dorsal vertebrae that ranging from mild to moderate. At the cutting vertebrae it was visible deformed and showed focal caseous content near the spinal cord, this was collected for microbiology where it was identified Salmonella sp. At microscopic evaluation the vertebrae had one to multifocal moderate inflammatory infiltrate of macrophages and heterophils. Some areas had lots of granulomas with central calcification and numerous giant cells. Other vertebras showed areas of osteomalácea and fibrosis. Rare focus had vertebral body fracture and spinal cord compression with mild infiltration entering the spinal cord canal. In the lung, especially in the bronchial epithelium, sometimes even within lymphocytes in bronchial-associated lymphoid tissue, in the intestine, liver, gall bladder, kidney and brain were found various structures of eosinophilic intracytoplasmic rounded ranging between 1 and 10 micrometers. These structures accompanied or not mononuclear inflammation. These findings are consistent with IBD and spondylitis due to salmonellosis. The IBD is a common disease in captive snakes, of world importance, is probably underdiagnosed in Brazil. This disease causes immunosuppression favoring the development of other affections, and is typically associated with other diseases such as spondylitis found in the case.(AU)

Detection of nidoviruses in live pythons and boas. / Nachweis von Nidoviren bei lebenden Pythons und Boas.

OBJECTIVES: Nidoviruses have recently been described as a putative cause of severe respiratory disease in pythons in the USA and Europe. The objective of this study was to establish the use of a conventional PCR for the detection of nidoviruses in samples from live animals and to extend the list of susceptible species. MATERIALS AND METHODS: A PCR targeting a portion of ORF1a of python nidoviruses was used to detect nidoviruses in diagnostic samples from live boas and pythons. A total of 95 pythons, 84 boas and 22 snakes of unknown species were included in the study. Samples tested included oral swabs and whole blood. RESULTS: Nidoviruses were detected in 27.4% of the pythons and 2.4% of the boas tested. They were most commonly detected in ball pythons (Python [P.] regius) and Indian rock pythons (P. molurus), but were also detected for the first time in other python species, including Morelia spp. and Boa constrictor. Oral swabs were most commonly tested positive. CONCLUSION: The PCR described here can be used for the detection of nidoviruses in oral swabs from live snakes. These viruses appear to be relatively common among snakes in captivity in Europe and screening for these viruses should be considered in the clinical work-up. CLINICAL RELEVANCE: Nidoviruses are believed to be an important cause of respiratory disease in pythons, but can also infect boas. Detection of these viruses in live animals is now possible and can be of interest both in diseased animals as well as in quarantine situations.