Draft assembly and annotation of the Cuban crocodile (Crocodylus rhombifer) genome.

OBJECTIVES: The new data provide an important genomic resource for the Critically Endangered Cuban crocodile (Crocodylus rhombifer). Cuban crocodiles are restricted to the Zapata Swamp in southern Matanzas Province, Cuba, and readily hybridize with the widespread American crocodile (Crocodylus acutus) in areas of sympatry. The reported de novo assembly will contribute to studies of crocodylian evolutionary history and provide a resource for informing Cuban crocodile conservation. DATA DESCRIPTION: The final 2.2 Gb draft genome for C. rhombifer consists of 41,387 scaffolds (contigs: N50 = 104.67 Kb; scaffold: N50-518.55 Kb). Benchmarking Universal Single-Copy Orthologs (BUSCO) identified 92.3% of the 3,354 genes in the vertebrata_odb10 database. Approximately 42% of the genome (960Mbp) comprises repeat elements. We predicted 30,138 unique protein-coding sequences (17,737 unique genes) in the genome assembly. Functional annotation found the top Gene Ontology annotations for Biological Processes, Molecular Function, and Cellular Component were regulation, protein, and intracellular, respectively. This assembly will support future macroevolutionary, conservation, and molecular studies of the Cuban crocodile.

The use of a penetrative captive bolt device during the killing of farmed saltwater crocodiles (Crocodylus porosus).

Killing of farmed saltwater crocodiles involve stunning with a penetrative captive bolt device, followed by a cut across the nape of the neck and physical destruction of the brain to ensure death. This study was a welfare-based assessment of the use of a penetrative captive bolt device in saltwater crocodiles, to determine whether it satisfies the criteria of a humane stunning method and can be regarded as a direct killing method without the need for the application of an adjunct method. Methods used were electroencephalogram (EEG), observation of post-stun behavior, and postmortem examination of gross pathology of the cranium and brain. 30 of 30 animals, demonstrated immediate and irreversible loss of consciousness. There was extensive damage to the brain in all animals, deemed to be inconsistent with cortical function and possible recovery. The CASH Special 0.22 penetrative captive bolt pistol (1.25 grain cartridge), applied to the top of the cranial plate, produced immediate and irreversible unconsciousness in all the animals studied. This method satisfies animal welfare expectations, providing crocodile processors with a technique that contributes to a humane killing process.

A true facultative carnivore? Effects of replacing ground chicken carcasses with soybean meal on the growth of captive broad-snouted caiman (Caiman latirostris) and its economics implications.

A specific diet for broad-snouted caiman, Caiman latirostris has not been designed despite the value of farm-raised caiman as an aquaculture product. To fill this gap, the objectives of this study were to evaluate the performance dietary replacement of ground chicken carcasses by of soybean meal (SM) as diet complement for C. latirostris. We conducted a 3-month growth trial to determine effects of graded levels of dietary SM on caiman growth as measured by increase in body length, body weight gain, food consumption (FC) and food conversion rate (FCR). Forty-eight hatchling caimans were fed with diets, composed primarily of practical feed ingredients, with 0, 25, or 40% dietary SM. Diets were fed three times per week for 90 days under temperature controlled. Body lengths and body weights were measured at 30-day intervals, and FC samples were taken between the 31-60-day interval. The results of this study indicate that the inclusion of SM in the diet of C. latirostris at levels of 25% increases in body length, body weight gain, FC and improve the FCR indicated that a concentration of 25% dietary SM (as fed) was adequate for growth of caimans under the conditions of this study. Results suggest that SM have a real nutritional contribution in the diet of broad-snouted caiman and can be used as an ingredient of the diet of the crocodilians raised in captivity, reducing production costs for sustainable use and conservation programs of this species.

New records of Hepatozoon and Oswaldofilaria from saltwater crocodiles (Crocodylus porosus) in Australia.

Diseases affecting wild Australian saltwater crocodiles (Crocodylus porosus) are rarely reported due to the difficulty in capturing animals and obtaining samples. In this investigation, we identified two haemoparasites (Hepatozoon and a filarial nematode) in saltwater crocodiles in Darwin, Australia. Light microscopic examination identified Hepatozoon in 7/7 (100%) wild crocodiles and in 2/20 (10%) of captive ones. When genomic DNAs from these same samples were further investigated using polymerase chain reaction (PCR)-based sequencing, we detected Hepatozoon in all 27 blood samples. Using both microscopy and PCR-based sequencing, we detected a filarial worm (proposed to be Oswaldofilaria) in one of 20 captive crocodiles. The sequence data were compared with sequence data available in public databases, and phylogenetic analyses indicated that the operational taxonomic units of Hepatozoon and Oswaldofilaria discovered here in these crocodiles are likely new species. This study is the first to use molecular tools to explore haemoparasites in Australian saltwater crocodiles and highlights the importance of health investigations in poorly studied vertebrate hosts.

Diversity and genetic characterization of Chlamydia isolated from Siamese crocodiles (Crocodylus siamensis).

Chlamydiosis, an infection caused by several Chlamydia species, has been reported in Nile, saltwater, and Siamese crocodiles. Despite its widespread reports in various countries, including Thailand, genetic information on Chlamydia species remains limited. This study presents a whole-genome-based characterization of Siamese crocodile-isolated Chlamydia. The results showed that Siamese crocodile Chlamydia contained a single circular chromosome with a size of 1.22-1.23 Mbp and a plasmid with a size of 7.7-8.0 kbp. A plasmid containing eight coding sequences (CDSs) was grouped in a ß lineage. A chromosome sequence had approximately 1,018-1,031 CDSs. Chlamydial factors involving virulence were documented in terms of the presence of cytotoxins and several virulence factors in the chromosomes of Siamese crocodile Chlamydia. The analysis of antimicrobial resistance genes in the Chlamydia genome revealed that the most common resistance genes were associated with aminoglycosides, fluoroquinolones, macrolides, tetracyclines, and cephalosporins, with loose matching (identities between 21.12 % and 74.65 %). Phylogenetic analyses, encompassing the assessments of both whole proteome and nine taxonomic markers, revealed that Siamese crocodile Chlamydia was separated into three lineages (lineages I-III) with high bootstrapping statistic support. Interestingly, isolate 12-01 differed genetically from the others, suggesting that it is a new member of Chlamydia. The study findings indicate that Siamese crocodiles are susceptible hosts to Chlamydia, involving more than one species. This study is the first employing the highest number of whole-genome data on Siamese crocodile Chlamydia and provides better insights into pathogen genetics.

Gelation of crocodile myofibrillar protein - κ-carrageenan mixtures in two low-NaCl solution.

Crocodile meat is a novel reptile meat source, but its processing method is rare. This study investigated the effect of κ-carrageenan addition and partial substitution of NaCl on the gel properties of crocodile myofibrillar protein (CMP). Result showed that CMP formed gel when temperature above 60 ℃. The water-holding capacity, gel strength, denaturation degree, sulfhydryl content covalent bond and hydrophobic bond of gel in KCl solution were significantly higher than those in CaCl2 solution (P < 0.05). K+ induced CMP to form a tight network structure with uniform small pores though covalent and hydrophobic bonds, but the gel properties were reduced by κ-carrageenan. In CaCl2 solution, κ-carrageenan improved the gel structure by filling the protein network through hydrogen bonding. Therefore, it can be concluded that KCl is better than CaCl2 in the manufacturing of low-sodium crocodile foods. Moreover, κ-carrageenan was only beneficial to gel quality in CaCl2 solution.

The Spatial Ecology of Nuisance Crocodiles: Movement Patterns of Relocated American Crocodiles (Crocodylus acutus) in Guanacaste, Costa Rica.

Anthropogenic alterations of the environment have increased, highlighting the need for human-wildlife coexistence and conflict mitigation. Spatial ecology, and the use of passive satellite movement technology in particular, has been used to identify patterns in human-wildlife conflict as a function of shared resources that present potential for dangerous situations. Here, we aim to remotely identify patterns indicative of human-crocodile conflict in Guanacaste, Costa Rica by exploring site fidelity and diverse modes of movement (i.e., land and water) across space between nuisance (relocated) and non-nuisance (wild) crocodiles. Advanced satellite remote sensing technology provided near-constant movement data on individuals at the regional scale. Telonics Iridium SeaTrkr-4370-4 transmitters were used with modified crocodilian fitting. Results indicate that relocated crocodiles exhibited large-scale movements relative to wild crocodiles. Nuisance relocated crocodiles either returned to the area of nuisance or potentially attempted to in short time frames. The results presented here highlight the need for alternative management strategies that facilitate relocation efficacy.

Enhancing microbial diversity as well as multi-organ health in hind-limb unloaded mice.

During space travel, the gut microbiota is changed which can lead to health-related issues. Previously, we utilized the hind-limb unloaded (HU) mouse, which is an established ground-based in-vivo model of microgravity and observed altered gut microbiota. In this study, we evaluated the beneficial effects of novel bacterial conditioned media in HU mice to understand if they can offset the effects of unloading in the HU mouse model. We aimed to explore the influence of bacterial conditioned media on diversity and quantity of intestinal microbes in HU mice, and investigated the microarchitecture of mice retinas and kidneys to evaluate the potential systemic effects of bacterial conditioned media in HU mice. Four-month-old, male C57/Bl6 mice were separated into groups: including the ground-based control group, the HU group mice fed with vehicle as placebo (HU-placebo mice), and the HU group fed with bacterial conditioned media (HU-CP mice) and kept under controlled environmental conditions for three weeks. Next, mice were sacrificed; gut dissections were conducted, and metagenomic analysis of bacterial species was performed via DNA extraction and 16S rRNA analysis. The results revealed an HU-induced reduction in intestinal microbial diversity, and an increase in pathogenic bacteria dominated by Firmicutes (45%). In contrast, supplementation with bacterial conditioned media for three weeks led to a significant increase in gut microbial diversity with noticeable changes in the OTUs abundance in the HU mice. Additionally, HU-induced muscle weakness and structural abnormalities in the retina and kidney were partially prevented with bacterial conditioned media. Moreover, a greater diversity of several bacteria in the HU-CP was observed including, Bacteriodota, Firmicutes, Proteobacteria, Actionobacteriota, Verrucomicorbiota, Cyanobacteria, Gemmatimonadota, Acidobacteriota, Chloroflexi, Myxococcota, and others. Prospective research involving molecular mechanistic studies are needed to comprehend the systemic effects of bacterial metabolites conditioned media on experimental animal models under chronic stress.

Underreported and taxonomically problematic: characterization of sanguinicolid larvae from freshwater limpets (Burnupiidae), with comments on the phylogeny and intermediate hosts of sanguinicolids.

Blood flukes of freshwater fish are understudied worldwide. Consequently, genetic information and data on their intramolluscan stages are scarce. In the current study, freshwater limpets of the genus Burnupia (Burnupiidae) from South Africa were examined for digeneans. Of 1645 specimens, 3.10% were infected by Sanguinicolidae larvae. Four sanguinicolids were distinguished by body size, number of penetration glands, tegumental spines' patterns and relative sizes of the finfolds on the body and furcae. Analyses of 28S, 18S and ITS rDNA sequences showed that the morphotypes were distinct from each other and from sanguinicolids whose genetic data are available. The present study is the first genetic characterization of sanguinicolids from Africa. Phylogenetic reconstruction revealed that the present species clustered with a sanguinicolid from Poland and were sister to Sanguinicola and Pseudosanguinicola from Russia and USA, respectively. The results indicate that the current species represent an unknown genus. What is more, blood fluke sequences from East Africa (presumed to be sanguinicolids), were distant from Sanguinicolidae and showed a closer relationship with acipensericolids from the USA. Freshwater fish blood flukes seem to be more diverse than previously recorded and use species of at least 13 gastropod families as intermediate hosts.

Muscle forces acting on the greater trochanter lead to a dorsal warping of the apophyseal growth plate.

The apophyseal growth plate of the greater trochanter, unlike most other growth plates of the human body, exhibits a curved morphology that results in a divergent pattern resembling an open crocodile mouth on plain antero-posterior radiographs. To quantify the angular alignment of the growth plate and to draw conclusions about the function of the muscles surrounding it, we analyzed 57 MRI images of 51 children and adolescents aged 3-17 years and of six adults aged 18-52 years. We measured the angulation of the plate relative to the horizontal plane (AY angle) and the trajectories of the muscles attaching to the greater trochanter of the proximal femur. From anterior to posterior, the AY angle shows a decrease of 33.44°. In the anterior third, the cartilage is angled at a mean of 51.64°, and in the posterior third, the mean angulation is 18.6°. This indicates that the cartilage in the anterior region of the greater trochanteric apophysis is subject to more vertically oriented force vectors compared to the posterior region, as the growth plates align perpendicular to the force vectors acting on them. Combining the measured muscle trajectories with the physiological cross-sectional areas (PCSA) available from the literature revealed that, in addition to the known internal and external lateral traction ligament systems, a third, dorsally located traction ligament system exists that may be responsible for the dorsal deformation of the AY angle.

Transesophageal echocardiography of cardiac function in Nile crocodiles - A novel tool for assessing complex hemodynamic patterns.

BACKGROUND: The crocodilian heart is unique among reptiles with its four-chambered structure and complete intracardiac separation of pulmonary and systemic blood flows and pressures. Crocodiles have retained two aortic arches; one from each ventricle, that communicate via Foramen of Panizza, immediately distally from the aortic valves. Moreover, crocodiles can regulate vascular resistance in the pulmonary portion of the right ventricular outflow tract (RVOT). These unique features allow for a complex regulation of shunting between the pulmonary and systemic circulations. Studies on crocodile shunting have predominantly been based on invasive measurements, but here we report on the use of echocardiography. METHODS: Experiments were performed on seven pentobarbital anaesthetized juvenile Nile crocodiles (length and mass of 192 ± 13 cm and 26 ± 5 kg, respectively). Echocardiographic imaging was performed using a transesophageal (TEE) approach. All images were EKG-gated. RESULTS: We obtain excellent views of cardiac structures and central vasculature through the esophagus. Standard imaging planes were defined for both long- and short axis views of the left ventricle and truncus arteriosus. For the RV, only a short axis view could be obtained. Color Doppler was used to visualize flow. Pulsed waved Doppler for measuring flow profiles across the atrioventricular valves, in the two RVOTs and the left ventricular outflow tract. Shunting across the Foramen of Panizza could be visualized and gated to the EKG. CONCLUSION: TEE can be used to image the unique features of the crocodile heart and allow for in-vivo imaging of the complex shunting hemodynamics, including timing of cardiac shunts.

Preserving Pure Siamese Crocodile Populations: A Comprehensive Approach Using Multi-Genetic Tools.

Hybrids between the critically endangered Siamese crocodile (Crocodylus siamensis) and least-concern saltwater crocodile (C. porosus) in captive populations represent a serious challenge for conservation and reintroduction programs due to the impact of anthropogenic activities. A previous study used microsatellite and mitochondrial DNA data to establish the criteria for identifying species and their hybrids; however, the results may have been influenced by biased allelic frequencies and genetic drift within the examined population. To overcome these limitations and identify the true signals of selection, alternative DNA markers and a diverse set of populations should be employed. Therefore, this study used DArT sequencing to identify genome-wide single nucleotide polymorphisms (SNPs) in both species and confirm the genetic scenario of the parental species and their hybrids. A population of saltwater crocodiles from Australia was used to compare the distribution of species-diagnostic SNPs. Different analytical approaches were compared to diagnose the level of hybridization when an admixture was present, wherein three individuals had potential backcrossing. Approximately 17.00-26.00% of loci were conserved between the Siamese and saltwater crocodile genomes. Species-diagnostic SNP loci for Siamese and saltwater crocodiles were identified as 8051 loci and 1288 loci, respectively. To validate the species-diagnostic SNP loci, a PCR-based approach was used by selecting 20 SNP loci for PCR primer design, among which 3 loci were successfully able to differentiate the actual species and different hybridization levels. Mitochondrial and nuclear genetic information, including microsatellite genotyping and species-diagnostic DNA markers, were combined as a novel method that can compensate for the limitations of each method. This method enables conservation prioritization before release into the wild, thereby ensuring sustainable genetic integrity for long-term species survival through reintroduction and management programs.

Placing Locking Compression Plates as an External Fixator in Wild Animal (Crocodile) Bite Victim: A Case Report.

Animal bites can cause severely contaminated open fractures, especially in the hand, wrist, and lower extremities, requiring expert orthopedic care. This case report emphasizes the unique issues posed by such injuries, particularly in low-income areas with limited access to comprehensive medical services. Pediatric examples, such as open tibiofibular fractures caused by animal encounters, highlight the significance of individualized treatment techniques. Crocodile bites, though infrequent, present an extreme peril and potential fatality. In Sri Lanka, where various crocodile species inhabit the waters, such incidents are rare but present. Long bone fractures caused by crocodile bites are significant and complicated injuries. The enormous force of a crocodile's jaw can cause substantial damage to bones, tendons, and surrounding tissues, often resulting in significant bone and soft tissue loss. Managing such injuries is extremely difficult, especially in low-resource settings. The use of a locking plate as an external fixator is a novel approach in the treatment of open fractures, nonunion, septic arthritis, and even as a distraction osteogenesis adjuvant. While it is not as common in typical fracture fixation textbooks, it is critical in specialized instances. This study describes a unique case of a 13-year-old boy with a Gustilo-Anderson Type IIIA crocodile bite who was treated in a low-income environment with a locking plate as an external fixator.

Play behavior in ectothermic vertebrates.

Until a few decades ago, play was considered a behavior unique to birds and mammals. Although play in other vertebrates is still a neglected research subject, data on it has been slowly accumulating, and are reviewed here. Now we know that animals as diverse as stingrays, cichlid fishes, monitor lizards, softshell turtles, and crocodiles can be unexpectedly playful. This knowledge has implications to broader theoretical problems, but much more research attention is needed to utilize its potential.

Structural variations in crocodile lizard populations provide novel insights into genomic variations in endangered animals.

Population-scale genome resequencing of endangered animals may contribute to gaining an understanding of how genomes vary as population sizes become smaller, as well as the functional implications of such variation. In this study, we analysed structural variations and gene presence and absence variations in the genomes of population of the endangered crocodile lizards. We found that the frequencies of some genes showed significant differences between crocodile lizards in different regions, indicating the influence of environmental selection, as well as potential contributions from demography and isolation, in shaping gene presence and absence variations. The haplotype diversity of major histocompatibility complex (MHC) genes was also found to differ among crocodile lizards inhabiting different regions. These findings indicate that well-designed interbreeding of crocodile lizards from different regions may facilitate the exchange of genes between different lizard populations and increase the haplotype diversity of MHC genes, which may be beneficial for the survival of these lizards. Our findings in this study, based on differences in gene structural variation, provide new insights into genomic variation and may contribute to the conservation of endangered animals.

Crocodile tears syndrome after vestibular schwannoma surgery: A case report.

INTRODUCTION: Crocodile tears syndrome is paroxysmal lacrimal hypersecretion while eating or drinking. It is relatively rare and mainly observed after peripheral facial nerve injury due to any cause. PRESENTATION OF CASE: We describe a case of crocodile tears syndrome after vestibular schwannoma surgery in a patient with pre-operative trigeminal nerve involvement. We also discuss the various treatment options offered to the patient and the natural course of the illness. DISCUSSION: It is hypothesized that facial nerve fibers undergo misdirected re-innervation of the lacrimal gland instead of the salivary gland. After vestibular schwannoma surgery, facial nerve palsy is well described. The sensory symptoms are relatively neglected by the surgeon, even though they cause discomfort to the patient. This may be the cause for the underestimation of its incidence. CONCLUSION: The knowledge of the incidence, mechanism and natural course of crocodile tears syndrome along with patient education is important to reduce post-operative distress to the patient. We also hypothesize the correlation of crocodile tears syndrome with motor component of facial function and the role of preservation of nervus intermedius in reducing post-operative facial palsy.

A masked gene concealed hand in glove in the forkhead protein crocodile regulates the predominant detoxification CYP6DA1 in Aphis gossypii Glover.

Cytochrome P450-mediated metabolism is an important mechanism of insecticide resistance, most studies show upregulated transcript levels of P450s in resistant insect strains. Our previous studies illustrated that some upregulated P450s were associated with cyantraniliprole resistance, and it is more comprehensive to use the tissue specificity of transcriptomes to compare resistant (CyR) and susceptible (SS) strains. In this study, the expression profiles of P450s in a CyR strain compared with a SS strain in remaining carcass or midgut were investigated by RNA sequencing, and candidate genes were selected for functional study. Drosophila melanogaster bioassays suggested that ectopic overexpression of CYP4CK1, CYP6CY5, CYP6CY9, CYP6CY19, CYP6CZ1 and CYP6DA1 in flies was sufficient to confer cyantraniliprole resistance, among which CYP6DA1 was the predominant contributor to resistance (12.24-fold). RNAi suppression of CYP4CK1, CYP6CY5, CYP6CY9 and CYP6DA1 significantly increased CyR aphid sensitivity to cyantraniliprole. The CYP6DA1 promoter had two predicted binding sites for crocodile (CROC), an intron-free ORF with bidirectional transcription yielding CROC (+) and CROC (-). Y1H, RNAi and EMSA found that CROC (-) was a transcription factor directly regulating CYP6DA1 expression. In conclusion, P450 genes contribute to cyantraniliprole resistance, and the transcription factor CROC (-) regulates the expression of CYP6DA1 in A. gossypii.

Gut microbiome of Crocodylus porosus and cellular stress: inhibition of nitric oxide, interleukin 1-beta, tumor necrosis factor-alpha, and prostaglandin E2 in cerebrovascular endothelial cells.

Crocodiles are renowned for their resilience and capacity to withstand environmental stressors, likely influenced by their unique gut microbiome. In this study, we determined whether selected gut bacteria of Crocodylus porosus exhibit anti-inflammatory effects in response to stress, by measuring nitric oxide release, interleukin 1-beta, tumor necrosis factor-alpha, and prostaglandin E2 in cerebrovascular endothelial cells. Using the Griess assay, the findings revealed that among several C. porosus gut bacterial isolates, the conditioned media containing the metabolites of two bacterial strains (CP27 and CP36) inhibited nitric oxide production significantly, in response to the positive control, i.e., taxol-treatment. Notably, CP27 and CP36 were more potent at reducing nitric oxide production than senloytic compounds (fisetin, quercetin). Using enzyme linked immunosorbent assays, the production of pro-inflammatory cytokines (IL-1ß, TNF-α, PGE2), was markedly reduced by treatment with CP27 and CP36, in response to stress. Both CP27 and CP36 contain a plethora of metabolites to exact their effects [(3,4-dihydroxyphenylglycol, 5-methoxytryptophan, nifedipine, 4-chlorotestosterone-17-acetate, 3-phenoxypropionic acid, lactic acid, f-Honaucin A, l,l-Cyclo(leucylprolyl), 3-hydroxy-decanoic acid etc.], indicative of their potential in providing protection against cellular stress. Further high-throughput bioassay-guided testing of gut microbial metabolites from crocodiles, individually as well as in combination, together with the underlying molecular mechanisms, in vitro and in vivo will elucidate their value in the rational development of innovative therapies against cellular stress/gut dysbiosis.

Plastisphere characterization in habitat of the highly endangered Shinisaurus crocodilurus: Bacterial composition, assembly, function and the comparison with surrounding environment.

Plastisphere is a new niche for microorganisms that complicate the ecological effects of plastics, and may profoundly influence biodiversity and habitat conservation. The DaGuishan National Nature Reserve, one of the largest habitats of the highly endangered crocodile lizard (Shinisaurus crocodilurus), is experiencing plastic pollution without sufficient attention. Here, plastisphere collected from captive tanks of crocodile lizards in this nature reserve was characterized for the first time. Three types of plastic (PE-PP, PE1, and PE2) together with the surrounding water and soil samples, were collected, and 16S rRNA sequencing technology was used to characterize the bacterial composition. The results demonstrated that plastisphere was driven by stochastic process and had a distinct bacterial community with higher diversity than that in surrounding water (p < 0.05). Bacteria related to nitrogen and carbon cycles (Pseudomonas psychrotolerans, Methylobacterium-Methylorubrum) were more abundant in plastisphere than in water or soil (p < 0.05). More importantly, plastics recruited pathogens and those bacteria function in antibiotic resistant genes (ARGs) coding. Bacteria related to polymer degradation also proliferated in plastisphere, especially Bacillus subtilis with a fold change of 42.01. The PE2 plastisphere, which had the lowest diversity and was dominated by Methylobacterium-Methylorubrum differed from PE 1 and PE-PP plastispheres totally. Plastics' morphology and aquatic nutrient levels contributed to the heterogeneity of different plastispheres. Overall, this study demonstrated that plastispheres diversify in composition and function, affecting ecosystem services directly or indirectly. Pathogens and bacteria related to ARGs expression enriched in the plastisphere should not be ignored because they may threaten the health of crocodile lizards by increasing the risk of infection. Plastic pollution control should be included in conservation efforts for crocodile lizards. This study provides new insights into the potential impacts of plastisphere, which is important for ecological risk assessments of plastic pollution in the habitats of endangered species.

Microplastic abundance, distribution, and diversity in water and sediments along a subtropical river system.

Microplastic pollution in aquatic environments has been a ubiquitous concern in recent years, owing to their rapid production combined with poor waste management practices. However, information on freshwater microplastics in the Global South is still scarce, despite growing research on freshwater microplastics in recent years, particularly within the Global North. To help address this knowledge gap, we studied water and sediment microplastic dynamics along a subtropical river system, i.e. Crocodile River around the Nelspruit City area (South Africa), across three different seasons (i.e. cool-dry, hot-dry, hot-wet) using a combination of diversity indices and multivariate analyses. Microplastics were more abundant during the cool-dry season in the surface water samples (mean 1058 particles m-3) and high during the hot-dry season (mean 568 particles kg-1 dwt) in the sediment samples. The hot-wet season had a low particle density in both surface water (mean 625 particles m-3) and sediments (mean 86 particles kg-1 dwt) samples. Microplastic shapes were dominated by fibres and fragments, with the colour scheme dominated by transparent, blue, and black. The abundance of microplastics was positively correlated with pH and resistivity, and negatively with river flow. Wastewater was attributed as a primary source of microplastics, particularly because of the observed dominant fibre microplastics, usually released during laundry. Our results suggest that Crocodile River and its tributaries are temporary sinks of microplastics during periods of low rainfall. Implications of this pollution are far-reaching, including effects on residents who are dependent on the Crocodile River as a source of drinking water and aquatic biota which may be exposed to these pollutants.