Histology of the chameleon liver

Histology of the chameleon liver

Tiermedizin Science

Histological examinations of organ tissue are part of every pathological examination in veterinary medicine. They are also frequently carried out in reptiles, but there are few studies on the histology of healthy organ tissue. An Arabic publication now deals with histological sections of chameleon livers.

Seven adult Yemen chameleons were captured in Abha City in the Aseer region and then killed with ether inhalation. The livers were placed in formalin and then poured into paraffin to make sections.

Morphologically, the liver was found to be a two-lobed, dark brown organ approximately 3.7 x 2 cm in size, which lies in the coelomic cavity in front of the stomach and surrounds the gall bladder. As in other animals, a capsule of connective tissue surrounds the liver.

Histologically, the liver of Yemen chameleons resembles that of other vertebrates in many respects. The liver capsule consists of closely spaced collagenous fibres and smooth muscle fibres. Normally, trabecular connective tissue divides the liver itself into many small lobules, but such a structure does not appear to be present in Yemen chameleons. In contrast to mammals, the liver cells (hepatocytes) are not arranged radially around a vein, but rather irregularly in follicles or alveoli. The hepatocytes are surrounded by capillary blood vessels. So-called melanoma macrophages, which are not found in birds and mammals, can be seen in the blood vessels. The hepatocytes in the Yemen chameleon are polyhedral or pyramid-shaped and usually contain several large, round cell nuclei in the periphery. The nuclei contain conspicuously dark nucleoli. Occasionally nuclei are central. Under haematoxylin-eosin (HE) staining, the hepatocytes appear very eosinophilic. In the connective tissue, branches of the portal vein, hepatic artery, small bile ducts and lymphatic vessels could be visualised. Haematopoietic tissue was found in the area directly under the liver capsule.

In addition to the histological examination, several pieces of liver were also examined using transmission electron microscopy. Images of both examination methods can be found in the publication.

Histomorphological, histochemical and ultrastructural studies on the healthy liver of Yemen Veiled Chameleon (Chamaeleo calyptratus) in Southern Saudi Arabia
Amin A. Al-Doaiss, Mohammed A. Alshehri, Ali A. Shati, Mohammad Y. Alfaifi, Mohammed A. Al-Kahtani, Ahmed Ezzat Ahmed, Refaat A. Eid, Laila A. Al-Shuraym, Fahd A. Al-Mekhlafi, Mohammed Al Zahrani, Mohammed Mubarak
International Journal of Morphology 41(5), 2023: pp. 1513-1526.
DOI: none

Image: Histological section of the liver of a Yemen chameleon from the above-mentioned publication

Genome of South African dwarf chameleons decoded

Genome of South African dwarf chameleons decoded

Science

After a reference genome for the panther chameleon (Furcifer pardalis) was recently published for the first time in China, scientists from South Africa have now followed with the genome of two dwarf chameleon species.

For the analyses, a male Bradypodion pumilum from Cape Town and a male Bradypodion ventrale from an introduced population in Johannesburg were taken. Muscle and liver tissue was used for long sequencing (HiC). The genome size of Bradypodion pumilum is 2.43 gigabase pairs (Gb), that of Bradypodion ventrale 2.40 Gb. The BUSCO analysis demonstrated a high completeness with about 97% of all existing coding genes in vertebrates. Furthermore, the current publication confirms the six macrochromosomes already found from the karyotype in Bradypodion thamnobates 2017. Various comparisons with Anolis sagrei were made. It remains open which chromosomes in Bradypodion are sex chromosomes.

The genomes can be viewed in the NCBI BioProject under the number PRJNA9861319 and under the BioSample numbers SAMN35825189 and SAMN35825190 respectively.

De novo whole genome assemblies for two Southern African Dwarf Chameleons (Bradypodion, Chamaeleonidae)
Jody M. Taft, Krystal A. Tolley, Graham J. Alexander, Anthony J. Geneva
Genome Biology and Evolution 15 (10), 2023, pp. 1-8
DOI: 10.1093/gbe/evad182

Findings on the synonyms of Trioceros ituriensis

Findings on the synonyms of Trioceros ituriensis

Verbreitung Science

Synonyms for the Congolese Ituri chameleon (Trioceros ituriensis) have existed for several decades. A recent publication by the herpetologist Wolfgang Böhme questions whether two of them could be separate species.

The US herpetologist Karl Patterson Schmidt described the chameleon as Chamaeleon ituriensis in 1919. At that time, Schmidt gave Medje, Ituri Forest, in the Democratic Republic of Congo as the type locality. He already noticed the external similarity to Chamaeleon johnstoni affinis, which is why he gave exactly that as a synonym of his Chamaeleon ituriensis. At the same time, Chamaeleon johnstoni affinis must not be confused with today’s Trioceros affinis, a separate species from Ethiopia that had already been described in 1845. In the course of the 20th century, Chamaeleo johnstoni affinis was placed in the genus Trioceros, sometimes thought to be a subspecies of its own, sometimes not. Böhme states that Trioceros johnstoni affinis is definitely a synonym of Trioceros ituriensis. Differences between Trioceros johnstoni and Trioceros ituriensis are the body size, the “reversed” sexual dimorphism (in T. ituriensis the females are larger than the males), a white line along the belly, several rows of enlarged scales along the side of the body, conical scales on the sides of the throat and the absence of rostral and preocular horns in male T. ituriensis.

However, the author is not sure about the species status of Chamaeleo laevigularis. The species was originally described from South Africa in 1926, then considered a synonym of Trioceros johnstoni and last identified as T. ituriensis by Tilbury in 2010. Böhme considers, because of different scaling of the throat, whether either a wrong locality was noted in the first description or it is a separate species, but could be extinct or lost.

Böhme also comes to the conclusion that Trioceros tremperi, which was described by Neĉas in 1994, could possibly also represent an already extinct species or a lost species and that the locality simply corresponded to incorrect information. Trioceros tremperi was last given as a synonym of Trioceros ituriensis by Tilbury 2010 and Spawls 2018. The chameleons had not been found in the type locality in Kenya before.

Documenting synonymies in Trioceros ituriensis (Schmidt, 1929) with remarks on sexual dimorphism in chameleons (Squamata: Chamaeleonidae)
Wolfgang Böhme
Revue Suisse de Zoologie 130(2), 2023: pp. 521-264
DOI: 10.35929/RSZ.0099

Photo: Trioceros ituriensis in the Budongo forest, Uganda, photographed by Katja Rembold

Effect of human chorionic gonadotropin in chameleons

Effect of human chorionic gonadotropin in chameleons

Tiermedizin Science

After a study on sperm collection in chameleons was already published this year, further results from the largely same team of authors now follow. The aim is to further research the basics of assisted reproduction, i.e. medical assistance in reproduction, in chameleons.

At Louisiana State University, 24 Veiled Chameleons were kept under standardised conditions for over a year. All animals came from a dealer who had taken them from the population of wild Veiled Chameleons in Florida. All were kept individually in ReptiBreeze, equipped with automatic sprinklers and artificial plants. Temperatures were around 28-29°C during the day with spots to seek higher values. 12 h UV-B irradiation per day was offered. Crickets and zophobas were fed. Before the start of the study, all 24 chameleons were clinically examined and several parasite treatments were carried out. Only after a month of acclimatisation did the actual study begin.

The first experiment tested what dose of human choriogonadotropin (hCG) is needed to increase the hormone levels of testosterone in the blood by 50%. Eleven Veiled Chameleons were randomly assigned to one of three groups. The three groups received injections of 100, 200 or 300 IU hCG under the skin at two-week intervals. Blood samples were taken before the first hormone injection and at 30 minutes, one hour, two hours, four hours, eight hours, 12 hours and 24 hours afterwards.

The second experiment tested the effect of hCG treatment on sperm production. 13 Veiled Chameleons were randomly assigned to a treatment or a control group. Once a week for one month, the animals in the first group were treated with 100 IU of hCG, while the second group was only injected with the same volume of isotonic saline. After a four-week break, the groups were switched and the experiment repeated. Blood samples to measure testosterone levels were taken before treatment and on day 15 and 30 afterwards. Semen was collected by electroejaculation under anaesthesia on the day before treatment and 30 days after.

The results showed that the testosterone level in male Veiled Chameleons increased significantly directly after the administration of hCG and remained elevated for about 24 hours. However, it did not matter which dose of hCG had been given beforehand. It could also be shown that the testosterone level increased significantly after the administration of hCG compared to the control group, which only received saline solution. The number of successful electroejaculations could be increased under hCG.

Effects of exogenous human chorionic gonadotropin administration on plasma testosterone and semen production in the Veiled Chameleon (Chamaeleo calyptratus)
Sean M. Perry, Sarah R. Camlic, Michael Lierz, Mark A. Mitchell
Journal of Herpetological Medicine and Surgery 33 (3), 2023, pp. 180-191
DOI: 10.5818/JHMS-D-22-00038

The microbiome of dwarf chameleons

The microbiome of dwarf chameleons

Tiermedizin Science

The term microbiome has been very popular for some years now. In humans and animals, it refers to the totality of all microorganisms that colonise a living being. Most of them colonise the gastrointestinal tract. In the case of chameleons, there is only very limited literature on this topic. A master’s thesis from South Africa now deals with the bacterial composition of the microbiome in South African dwarf chameleons of the genus Bradypodion.

60 cheek swabs were collected from wild chameleons in KwaZulu-Natal. Of these, 20 were cheek swabs from Bradypodion melanocephalum, 20 from Bradypodion thamnobates and 20 from Bradypodion setaroi. After sampling, the same 60 animals were transported in cloth bags to the research base, where the animals were kept in 3.3 l boxes for 24 hours to obtain faecal samples. Since not all of the original 60 chameleons defecated, faeces were collected from additional chameleons.

The samples were all genetically tested. 40.43% of the samples contained Firmicutes, a similarly large proportion of the samples contained Proteobacteria with 36.86%. Bacteroidota followed with some distance, which could be detected in just under 16% of the samples. Verrucomicrobiota, Fusobacteriota, Actinobateriota, Spirochetes, Desulfobacteroa, Cyanobacteria, Thermoplamatota, Deferribacterota, Synergistota, Campylobacterota, Deinococcota, Halobacterota, Euryarchaeota, Elusimicrobiota and Myxococcota were found in significantly smaller numbers (up to 2%).

The microbiome of dwarf chameleons of the species Bradypodion melanocephalum, Bradypodion thamnobates and Bradypodion setaroi is similar to that of other reptiles. It consists mainly of proteobacteria and firmicutes, which may contribute to digestion. One particular bacterial species also suggests that the diet of the studied dwarf chameleons may include beetles of the genus Dendrophagus. The microbiome of all three dwarf chameleon species was very similar in the cheek swabs – this is called phylosymbiosis – while there were differences in composition between the species in the faeces. In all three dwarf chameleon species, significantly more different bacteria were found in the faeces than in the cheek swabs. A comparison between males and females did not reveal any significant differences in the microbiome of all three chameleon species. The author assumes that the bacterial species depend on the different habitats of the respective species. It is still unclear to what extent the microbiome is related to bacteria that a chameleon may ingest with feeding insects or from the soil of its environment. A detailed list of the bacterial species found can be found in the appendix of the publication.

The Hitchhiker’s Guide to dwarf chameleons (Bradypodion): The composition and function of the microbiome
Matthew G. Adair
Master of Science dissertation at the university of Johannesburg, 2023
DOI: not available

Concerning Florida’s introduced panther chameleons

Concerning Florida’s introduced panther chameleons

Verbreitung Science

The “Sunshine State” Florida in the USA has the largest number of non-native species of reptiles in the world because of its warm climate. The panther chameleon (Furcifer pardalis) is one of the invasive species, i.e. those that do not actually belong in Florida but are now reproducing there. A study has now investigated the question of what the human inhabitants of Florida actually think of the chameleons.

It has been discussed for a long time whether panther chameleons belong to the species that were deliberately released for the purpose of “ranching”, i.e. to collect the offspring of the released chameleons for sale. That private individuals collect panther chameleons is not in dispute. According to the authors, ranching populations in Florida are mostly kept secret. They became aware of a small population in Orange County via social media in 2019. They then searched for the animals at night with torches and actually found 26 panther chameleons during several walks. They encountered private individuals on several occasions who were also looking for chameleons.

In 2020, questionnaires were distributed in person and via flyers with QR codes to 248 households located within the presumed 0.9 km² distribution area of the panther chameleon population. They were asked about concerns regarding the occurrence of panther chameleons, but also about existing knowledge about invasive species in general. The residents were also divided into three areas: A core region where chameleons had been observed several times, a peripheral region with few findings, and an outer region where no chameleons had been sighted at all.

44 households answered the questionnaire.  In fact, all 11 interviewed residents in the outer region had not sighted any chameleons. Of the 33 residents interviewed in the core and peripheral region, about a third said they had already observed panther chameleons. The same number had seen the light of torches at night. 86% of the residents surveyed knew that panther chameleons are not actually native to Florida. Only a few residents said they were concerned about the occurrence. Seven residents had approached collectors with torches and said the collectors had all said they were looking for chameleons for research purposes. Only one of the collectors had said he/she was looking for animals to sell, according to the residents. One resident reported an altercation after strangers entered his property several times looking for chameleons. Another resident called the police because of a whole group of collectors on the neighbouring property.

Unfortunately, the questionnaire was given out after the search efforts of the authors themselves, so it is not apparent from the responses how many of the encounters were indeed with people looking for chameleons for sale purposes. The publication is also a preprint, so no review process has taken place yet.

Colorful lizards and the conflict of collection
Colin M. Goodman, Natalie M. Claunch, Zachary T. Steele, Diane J. Episcopio-Sturgeon, Christina M. Romagosa
Preprint, 2023
DOI: 10.1101/2023.08.10.552819

Picture: Alex Laube

Unknown chameleon discovered in Ivohiboro forest (Madagascar)

Unknown chameleon discovered in Ivohiboro forest (Madagascar)

Verbreitung Science

There are still almost unexplored areas on Madagascar today. The Ivohiboro rainforest is located in the southeast of the island in the protected area of the same name, southwest of the southernmost foothills of the Andringitra Mountains. The forest itself is about 8.58 km² in size and thus only occupies a small part of the protected area. It is surrounded by savannahs and spans altitudes from 650 to 1460 m above sea level. The protected area is currently managed by local organisations and Madagascar’s Ministry of Environment. The last expedition to explore the Ivohiboro forest took place in 1924. Since 2016, researchers from the USA and Great Britain have now undertaken six expeditions to the small forest to study the biodiversity of plants, birds, mammals, reptiles, and amphibians there in more detail.

To detect reptiles and amphibians, the forest was divided into nine transects of about 200 x 20 m, each more than 200 m apart. The transects were searched for several days and nights. All animals found were documented and, if possible, identified down to genus or species level.

As a result, the scientists were able to identify 107 species of vertebrates and 219 plants. This enormous diversity of species underlines the importance of preserving the forest in terms of species conservation and indicates a well-functioning ecosystem. Among the species found were two chameleons: a Palleon species and a small Calumma. Unfortunately, the publication does not provide any further information on the former. The small Calumma had a conspicuous blue coloured rostral appendage, as it is found in Calumma linotum or Calumma boettgeri in the far north of Madagascar. As genetic studies are still lacking, it is unclear whether these chameleons are an extremely wide range extension – Ivohiboro lies about 1000 km south of the ranges of Calumma boettgeri and Calumma linotum – or whether it is perhaps even a new, as yet undescribed species.

A surprising haven: The biodiversity of an old-growth forest amidst a scorched landscape in Madagascar
Beatriz Otero Jimenez, Ren Montaño, Ryan S. Rothman, Rachel C. Williams, Patricia C. Wright
Conservation Science and Practice, 2023
DOI: 10.1111/csp2.12993

Genome of the panther chameleon decoded

Genome of the panther chameleon decoded

Science

In recent decades, genetic research has developed rapidly. Since 2009, the so-called high fidelity (HiFi) Pacbio sequencing method has been available for sequencing genomes. Nevertheless, relatively little is being done in the reptile field. There are only about a hundred so-called reference genomes for reptiles, and none at all for chameleons. Scientists from China have now published a reference genome for the panther chameleon (Furcifer pardalis).

For the analysis, a 5-year-old male captive panther chameleon was killed using isoflurane and then dissected. Different tissues were frozen in liquid nitrogen. Skeletal muscle was used for short genome DNA sequencing and HI-C sequencing. Liver was used for HiFi sequencing. RNA from heart, liver, spleen, testis, lung, kidney, and skin were used for transcriptome sequencing.

The genome size of the panther chameleon from the K-mer analysis is 1.61 gigabase pairs (Gbp), containing only 22 so-called contigs, sets of overlapping DNA. The karyotype contains 11 chromosomes, each consisting of one to four contigs. Ten out of eleven chromosomes have repeat sequences (TAACCC). BUSCO analysis demonstrated a high completeness of the genome. The genome can be viewed in the NCBI BioProject under the number PRJNA974816 and in ScienceDataBank.

Efficient and highly continuous chromosome-level genome assembly of the first chameleon genome
Hongxin Xie, Zixuan Chen, Shuai Pang, Weiguo Du
Genome Biology and Evolution 131, 2023
DOI: 10.1093/gbe/evad131

 

Picture: Alex Laube

Zoonotic potential of Yemen chameleons in Gran Canaria (Spain)

Zoonotic potential of Yemen chameleons in Gran Canaria (Spain)

Science

The Canary Islands are located northwest of Africa near the coast of Morocco. On Gran Canaria, the second largest island, around 290 of over 1000 plant and animal species have been introduced, i.e. species that do not originally occur there. Since at least 2017, there have been free-living Veiled chameleons (Chamaeleo calyptratus) on Gran Canaria. Spanish scientists have now investigated whether this introduced chameleon population could have zoonotic potential.

They examined 40 Veiled chameleons that had previously been caught and killed by Red de Alerta Temprana de Canarias para la Detección e Intervención de Especies Exóticas Invasoras in Arucas. 36 of the chameleons were adults, four were juveniles. Intestinal contents were taken from each chameleon and analysed for the presence of different bacteria using various methods.

At least one of the bacteria sought was found in 28 of the Veiled chameleons. About half of the chameleons had Yersinia enterocolitica, which is the highest prevalence ever recorded for this bacterium in reptiles. The bacterium can cause diarrhoea in humans, among other things. It is unclear how the Veiled chameleons became infected with it – possibly via insects. 16 of the Veiled chameleons had salmonella in their intestines. Salmonella is very common in reptiles and has even been found in endemic species on Gran Canaria. Pseudomonas is also frequently found in reptiles and was detected in the intestines of 13 animals. Two Veiled chameleons were infected with Campylobacter, in one of them Campylobacter lari could be identified. This bacterium can rarely cause illness in humans, but no pathogenic potential for humans is known for the species otherwise common in reptiles. Campylobacter lari has so far been detected mainly in seafood and birds – it is possible that the Veiled Chameleons picked up the bacterium on the coast and did not bring it with them. Three Veiled chameleons had Escherichia coli, which in rare cases can lead to haemolytic uraemic syndrome (HUS) in humans. Another two chameleons had Listeria monocytogenes in their intestines, which can be dangerous for pregnant women if ingested with food. Five Veiled Chameleons had mycobacteria, several of which were found to be non-tuberculous infections. Staphylococci were detected in seven chameleons, but they are part of the normal skin flora. However, five isolates were positive for resistance to certain antibiotics, which is becoming an increasing problem with Staphylococcus aureus in humans. Most recently, Vibrio was detected in a single Yemen chameleon, some species of which can cause diarrhoea in humans. The bacterium has previously been detected in introduced anoles on Tenerife.

The authors state that there is a zoonotic potential for humans due to the handling of introduced Veiled Chameleons on Gran Canaria. However, the extent to which there is a real risk for humans as well as endemic species needs to be further investigated.

Study of zoonotic pathogens in alien population of Veiled Chameleons (Chamaeleo calyptratus) in the Canary Islands (Spain)
Román Pino-Vera, Néstor Abreu-Acosta, Pilar Foronda
Animals 13 (14), 2023
DOI:  10.3390/ani13142288

Chameleons in the Montagne des Français (Madagascar)

Chameleons in the Montagne des Français (Madagascar)

Verbreitung Science

The Montagne des Français is a limestone massif with dry forest in northern Madagascar. It reaches up to 425 m above sea level and is within sight of the largest coastal town in the north, Antsiranana (French Diego Suarez). It has been a protected area since 2007. Scientists from Madagascar and the USA conducted counts of reptiles and amphibians in the Montagne des Français in 2014 and 2020.

Counts were made in January and May, i.e. during and at the end of the rainy season. In 2014, the focus was on the region around Andavakoera, while in 2020 it was on Sahabedara, Ampitiliantsambo, and Andavakoera. In order to find animals, the search was conducted during the day and at night along predefined paths, partly in suitable habitats, and partly in pitfall pits.

A total of 20 amphibian and 50 reptile species were recorded. Four new amphibians and one reptile were found for the first time in the Montagne des Français. The snake Langaha pseudoalluaudi was discovered again for the first time since 2007. Among the chameleons, there were minor new discoveries. Brookesia stumpffi could only be found in 2014, but no longer in 2020 – however, due to the relatively wide distribution of the species, this should not be a problem for the entire population. Brookesia tristis, one of the smallest chameleons in the world, was also only seen in 2014. Here, the body size, which makes it very difficult to find, and the time of year (May is relatively late for this species) could play a role. Brookesia ebenaui was detected in Andavakoera in 2014 and in Sahabedara in 2020. The two tree dwellers Furcifer oustaleti and Furcifer pardalis were found in both years in Andavakoera and Ampitiliantsambo. Furcifer petteri, on the other hand, was present at all the sites surveyed in both years.

Amphibians and reptiles of the “Montagne des Français”: Update of the distribution and regional endemicity
Herizo Oninjatovo Radonirina, Bernard Randriamahatantsoa, Rabibisoa Harinelina Christian Nirhy, Christopher J. Raxworthy
Preprint
DOI: 10.20944/preprints202306.1499.v1

Photo: Furcifer petteri on Madagascar, photographed by A. Laube