Development of an enzyme-linked immunosorbent assay for vitellogenin of Morelet's crocodile (Crocodylus moreletii)

The purpose of this study was to develop an immunoassay for vitellogenin in Morelet's crocodile (Crocodylus moreletii). Blood was collected from wild-caught crocodiles in Belize. Plasma samples from adult females taken during the breeding season were used for vitellogenin purification and samples from adult males were used for comparison. No differences were detected between males and females for plasma total protein concentration, as measured by Coomassie assay. However, denaturing polyacrylamide gel electrophoresis (SDS-PAGE) revealed that female plasma contained a 210-kDa protein, presumably the vitellogenin monomer, that was absent in adult male plasma. The identity of the putative vitellogenin was confirmed by its cross-reactivity in Western blots with a vitellogenin antiserum that was generated against a conserved vitellogenin peptide sequence. Crocodile vitellogenin was purified by two successive rounds of DEAE chromatography. The purified protein had an apparent molecular mass of 450 kDa, as determined by gel filtration chromatography, and 210 kDa on SDS-PAGE. An indirect enzyme-linked immunosorbent assay (ELISA) was then developed for C. moreletii vitellogenin. The detection limit of the assay was 20.0 ng/mL. The intra- and inter-assay coefficients of variation were 5.3% and 9.8%, respectively. The recovery of vitellogenin diluted into male plasma was 94.7%. The ELISA assay revealed that vitellogenin levels of adult female plasma during the breeding season ranged from 1.8 to 3.1 mg/mL with a mean of 2.5+/-0.25 mg/mL. No vitellogenin was detected in adult male plasma. Induction of vitellogenin in Morelet's crocodile may be a useful model system for field studies of crocodile reproduction and for investigations of endocrine disruption in this species.     

Effect of human-mediated migration and hybridization on the recovery of the American crocodile in Florida (USA)

The American crocodile (Crocodylus acutus) is a large apex predator with an extensive but fragmented range. Crocodylus acutus suffered a population crash and was almost extirpated from Florida (USA) during the 1970s; however, in 2007 it was federally downlisted from endangered to threatened based on an increase of suitable habitat and a growing population. A genetic assessment of this population has not been performed even though foreign crocodiles have been released into Florida waters. Herein, we use mitochondrial and nuclear DNA markers to characterize relationships among American crocodiles along the southern coast of Florida and reference samples from other countries. We detected inter- and intra-specific hybridization and unexpected population structure attributed to human-mediated migration of crocodiles from Latin America and the Greater Antilles. Our results suggest that the population size of crocodiles actually native to Florida should be reevaluated, particularly in light of ongoing admixture in this population. We reemphasize the utility of genetic markers in conservation and management programs for endangered species; especially those that can hybridize with closely related congeners. The American crocodile in Florida has recovered owing to successful conservation initiatives, but a long-term management protocol that takes genetic data into account is still needed.     

Organochlorine pesticides in chorioallantoic membranes of Morelet's crocodile eggs from belize

Recent studies examined the utility of the chorioallantoic membrane (CAM) as a nonlethal, noninvasive indicator of environmental contaminant exposure in oviparous wildlife. The CAM is a highly vascularized extraembryonic membrane that functions as a site for respiration, nutrient transport, and waste storage during embryonic development. After hatching, the CAM is usually discarded with the eggshell and can be used for chemical residue analysis. Chorioallantoic membranes have been used successfully to examine contaminant exposure and predict chemical concentrations in multiple species of birds and reptiles. In this study, we examined organochlorine (OC) pesticide concentrations in CAMs from eggs of Morelet's crocodiles (Crocodylus moreletii) from northern Belize. Multiple OCs were detected in crocodile CAMs, including aldrin, dieldrin, endrin, dichlorodiphenyltrichloroethane, dichlorodiphenyldichloroethane, dichlorodiphenyldichloroethylene (DDE), heptachlor, lindane, and methoxychlor. Number and concentrations of OC compounds in CAMs were variable. The most prevalent contaminant detected was DDE, which occurred in 69% of CAMs, with concentrations ranging from 0.3 parts per billion (ppb) to 17.0 ppb. The OC burdens in crocodile CAMs confirm contamination of eggs and suggest exposure in embryos and maternal females. These results further support the use of CAMs as qualitative indicators of OC exposure in oviparous wildlife. The efficacy of this sampling technique in the field will depend on the logistics and cost associated with CAM collection and the specific life history traits of the wildlife species.     

Diet and body condition of cave‐dwelling dwarf crocodiles (Osteolaemus tetraspis,Cope 1861) in Gabon

We present the first ever observations of dwarf crocodiles (Osteolaemus tetraspis) selectively utilizing a cave ecosystem. We analysed crocodile diet to preliminarily assess the degree of ecological isolation of cave‐dwelling crocodiles from their forest‐dwelling conspecifics despite their physical proximity. We additionally conducted the first study of body condition of dwarf crocodiles and compared cave‐dwelling individuals to surface‐dwelling individuals from a variety of alternative habitats and sites as a means to better understand the implications of a semihypogean life on this species. Crocodiles captured in the caves appeared to forage exclusively in the caves and ate mostly cave crickets and bats, whereas crocodiles captured in the surrounding forest habitat consumed primarily freshwater crustaceans and insects and were not found to consume cave‐dwelling prey. Juvenile cave crocodiles had significantly higher body condition compared to juvenile forest crocodiles, which did not vary amongst forest locations. The difference in body condition between adult cave and forest crocodiles was not statistically significant despite also being higher; we suspect this is an artefact of the low sample size rather than a real nondifference. Forest adult crocodiles generally had significantly higher body condition than juveniles, but did not vary by site or habitat. This lack of variation suggests that habitat type is not the most important factor influencing dwarf crocodile body condition. Our results provide a unique insight into facultative cave use by a principally surface‐dwelling species and reinforce the necessity for further research into this unique system to better understand the evolutionary‐scale implications of cave habitat use by dwarf crocodiles.     

A sampling design and model for estimating abundance of Nile crocodiles while accounting for heterogeneity of detectability of multiple observers

As part of the development of a management program for Nile crocodiles in Lake Nasser, Egypt, we used a dependent double-observer sampling protocol with multiple observers to compute estimates of population size. To analyze the data, we developed a hierarchical model that allowed us to assess variation in detection probabilities among observers and survey dates, as well as account for variation in crocodile abundance among sites and habitats. We conducted surveys from July 2008–June 2009 in 15 areas of Lake Nasser that were representative of 3 main habitat categories. During these surveys, we sampled 1,086 km of lake shore wherein we detected 386 crocodiles. Analysis of the data revealed significant variability in both inter- and intra-observer detection probabilities. Our raw encounter rate was 0.355 crocodiles/km. When we accounted for observer effects and habitat, we estimated a surface population abundance of 2,581 (2,239–2,987, 95% credible intervals) crocodiles in Lake Nasser. Our results underscore the importance of well-trained, experienced monitoring personnel in order to decrease heterogeneity in intra-observer detection probability and to better detect changes in the population based on survey indices. This study will assist the Egyptian government establish a monitoring program as an integral part of future crocodile harvest activities in Lake Nasser. © 2012 The Wildlife Society.     

Ectromelia in Morelet's crocodile from Belize

Two Morelet's crocodiles (Crocodylus moreletii) captured on 21 March 1997 and 20 April 1998 in the New River system, Belize exhibited ectromelia of one forelimb. External and radiograph examination appears to indicate limb agenesis of unknown etiology, as there is no apparent scarring or skeletal trauma. These two individuals represent the only cases of missing limbs from 642 individuals captured in this study and to our knowledge, the first reported cases in Morelet's crocodile. Several factors including age and diet of the reproducing female, extremes in nest conditions (egg incubation temperature and humidity), and exposure to environmental contaminants can cause developmental abnormalities in crocodilians and may have contributed to the condition observed in these animals. Survival rates for hatchling crocodilians are generally low, and embryonic malformations such as ectromelia may constitute an added disadvantage to survival. However, both individuals examined in this study were vigorous and appeared in good condition.     

A novel multiplex PCR assay for the identification of Indian crocodiles

Illegal hunting has been a major threat for the survival of wildlife fauna, including the three crocodile species that India harbours: Crocodylus palustris, Crocodylus porosus and Gavialis gangeticus. Although law prevents trade on these species, illicit hunting for trade continues to threaten the survival of these endangered species; conservation strategies therefore require a rapid molecular identification technique for Indian crocodiles. A multiplex polymerase chain reaction (PCR) assay with species-specific primers, considered as one of the most effective molecular techniques, is described herein. The primers were designed to yield species-specific sized amplicons. The assay discriminates the three Indian crocodile species unambiguously within a short time period using only simple agarose gel electrophoresis. We recommend this multiplex PCR assay to be used in the identification of Indian crocodile species.     

Effects of Different Rearing Strategies and Ages on Levels of Natural Antibodies in Saliva of the Philippine Crocodile

The endemic Philippine crocodile(Crocodylus mindorensis) is a relatively small, critically endangered freshwater crocodile. In a head start program, crocodile hatchlings are caught in the wild, reared in captivity, and released back into the wild after two years. The current study aimed to determine optimal rearing strategies of Philippine crocodile hatchlings, including identification of possible diseases during rearing, and studying the effect of ages on natural antibody(NAb) levels. Thirty Philippine crocodiles were divided into two groups, half were reared with a hiding board, and half without the hiding board. Both groups received three different kinds of diets: meat, shrimp, or a combination of both. Saliva samples of the crocodiles were taken three times over a period of three months to test for NAb levels. Saliva samples were also taken from older crocodiles and crocodiles from different locations. NAb titres were compared to sheep red blood cells. Each time saliva samples were taken, a health check was done. The results showed that crocodiles would prefer the hiding board, and neither housing nor diet could affect the level of NAb titres in saliva. A positive correlation was found between NAb titres and body size, weight and age. Wild hatchlings had higher NAb titres than the hatchlings born in captivity, but the difference diminished with ageing. Five different diseases were found.     

Proton leak in hepatocytes and liver mitochondria from archosaurs (crocodiles) and allometric relationships for ectotherms

It has previously been shown that mitochondrial proton conductance decreases with increasing body mass in mammals and is lower in a 250-g lizard than the laboratory rat. To examine whether mitochondrial proton conductance is extremely low in very large reptiles, hepatocytes and mitochondria were prepared from saltwater crocodiles ( Crocodylus porosus) and freshwater crocodiles ( Crocodylus johnstoni). Respiration rates of hepatocytes and liver mitochondria were measured at 37 degrees C and compared with values obtained for rat or previously measured for other species. Respiration rates of hepatocytes from either species of crocodile were similar to those reported for lizards and approximately one fifth of the rates measured using cells from mammals (rat and sheep). Ten-to-thirty percent of crocodile hepatocyte respiration was used to drive mitochondrial proton leak, similar to the proportion in other species. Respiration rates of crocodile liver mitochondria were similar to those of mammalian species. Proton leak rate in isolated liver mitochondria was measured as a function of membrane potential. Contrary to our prediction, the mitochondrial proton conductance of liver mitochondria from crocodiles was greater than that of liver mitochondria from lizards and was similar to that of rats. The acyl composition of liver mitochondrial phospholipids from the crocodiles was more similar to that in mitochondria from rats than in mitochondria from lizards. The relatively high mitochondrial proton conductance was associated with a relatively small liver, which seems to be characteristic of crocodilians. Comparison of data from a number of diverse ectothermic species suggested that hepatocyte respiration rate may decrease with body mass, with an allometric exponent of about -0.2, similar to the exponent in mammalian hepatocytes. However, unlike mammals, liver mitochondrial proton conductance in ectotherms showed no allometric relationship with body size.     

Antibacterial activity of plasma from crocodile (Crocodylus siamensis) against pathogenic bacteria

ABSTRACT: BACKGROUND: The Siamese crocodile (Crocodylus siamensis) is a critically endangered species of freshwater crocodiles. Crocodilians live with opportunistic bacterial infection but normally suffer no adverse effects. They are not totally immune to microbial infection, but their resistance thereto is remarkably effective. In this study, crude and purified plasma extracted from the Siamese crocodile were examined for antibacterial activity against clinically isolated, human pathogenic bacterial strains and the related reference strains. METHODS: Crude plasma was prepared from whole blood of the Siamese crocodile by differential sedimentation. The crude plasma was examined for antibacterial activity by the liquid growth inhibition assay. The scanning electron microscopy was performed to confirm the effect of crude crocodile plasma on the cells of Salmonella typhi ATCC 11778. Effect of crude crocodile plasma on cell viability was tested by MTT assay. In addition, the plasma was purified by anion exchange column chromatography with DEAE-Toyopearl 650M and the purified plasma was tested for antibacterial activity. RESULTS: Crude plasma was prepared from whole blood of the Siamese crocodile and exhibited substantial antibacterial activities of more than 40% growth inhibition against the six reference strains of Staphylococcus aureus, Salmonella typhi, Escherichia coli, Vibrio cholerae, Pseudomonas aeruginosa, and Staphylococcus epidermidis, and the four clinical isolates of Staphylococcus epidermidis, Pseudomonas aeruginosa, Salmonella typhi, and Vibrio cholerae. Especially, more than 80% growth inhibition was found in the reference strains of Salmonella typhi, Vibrio cholerae, and Staphylococcus epidermidis and in the clinical isolates of Salmonella typhi and Vibrio cholerae. The effect of the crude plasma on bacterial cells of Salmonella typhi, a certain antibacterial material probably penetrates progressively into the cytoplasmic space, perturbing and damaging bacterial membranes. The effect of the crude plasma was not toxic by the yellow tetrazolium bromide (MTT) assay using a macrophage-like cell, RAW 264.7. The pooled four fractions, designated as fractions D1-D4, were obtained by column chromatography, and only fraction D1 showed growth inhibition in the reference strains and the clinical, human pathogenic isolates. CONCLUSIONS: The crude and purified plasma from the Siamese crocodile significantly showed antibacterial activity against pathogenic bacteria and reference strains by damage cell membrane of target bacterial cells. From the MTT assay, the Siamese crocodile plasma was not cytotoxic to the cells.     

Dead or Alive? Factors Affecting the Survival of Victims during Attacks by Saltwater Crocodiles (Crocodylus porosus) in Australia

Conflicts between humans and crocodilians are a widespread conservation challenge and the number of crocodile attacks is increasing worldwide. We identified the factors that most effectively decide whether a victim is injured or killed in a crocodile attack by fitting generalized linear models to a 42-year dataset of 87 attacks (27 fatal and 60 non-fatal) by saltwater crocodiles (Crocodylus porosus) in Australia. The models showed that the most influential factors were the difference in body mass between crocodile and victim, and the position of victim in relation to the water at the time of an attack. In-water position (for diving, swimming, and wading) had a higher risk than on-water (boating) or on-land (fishing, and hunting near the water''s edge) positions. In the in-water position a 75 kg person would have a relatively high probability of survival (0.81) if attacked by a 300 cm crocodile, but the probability becomes much lower (0.17) with a 400 cm crocodile. If attacked by a crocodile larger than 450 cm, the survival probability would be extremely low (<0.05) regardless of the victim’s size. These results indicate that the main cause of death during a crocodile attack is drowning and larger crocodiles can drag a victim more easily into deeper water. A higher risk associated with a larger crocodile in relation to victim’s size is highlighted by children’s vulnerability to fatal attacks. Since the first recently recorded fatal attack involving a child in 2006, six out of nine fatal attacks (66.7%) involved children, and the average body size of crocodiles responsible for these fatal attacks was considerably smaller (384 cm, 223 kg) than that of crocodiles that killed adults (450 cm, 324 kg) during the same period (2006–2014). These results suggest that culling programs targeting larger crocodiles may not be an effective management option to improve safety for children.     

Cloacal and nasal bacterial flora of Lepidochelys olivacea (Testudines: Cheloniidae) from the North Pacific Coast of Costa Rica

Cloacal and nasal bacterial flora of Lepidochelys olivacea (Testudines: Cheloniidae) from the North Pacific coast of Costa Rica. The aerobic cloacal and nasal bacterial flora of 45 apparently healthy female olive ridley sea turtles (Lepidochelys olivacea) was studied at Nancite nesting beach, in Santa Rosa National Park (Costa Rican North Pacific) during July and August 2002. Bacterial samples were obtained by inserting sterile swabs directly into the cloaca and the nasal cavities of the turtles. Ninety-nine aerobic bacterial isolates, including 10 Gram-negative and 5 Gram-positive bacteria, were recovered. The most common bacteria cultured were Aeromonas spp. (13/45) and Citrobacter freundi (6/45) from cloacal samples and Bacillus spp. (32/45), Staphylococcus aureus (6/45) and Corynebacterium spp. (5/45) from nasal ducts. The results of the present study showed that the aerobic bacterial flora of nesting female olive ridleys was composed of several potential human and animal microbe pathogens.     

Determinants of habitat selection by hatchling Australian freshwater crocodiles

Animals almost always use habitats non-randomly, but the costs and benefits of using specific habitat types remain unknown for many types of organisms. In a large lake in northwestern Australia (Lake Argyle), most hatchling (<12-month-old) freshwater crocodiles (Crocodylus johnstoni) are found in floating vegetation mats or grassy banks rather than the more widely available open banks. Mean body sizes of young crocodiles did not differ among the three habitat types. We tested four potential explanations for non-random habitat selection: proximity to nesting sites, thermal conditions, food availability, and exposure to predation. The three alternative habitat types did not differ in proximity to nesting sites, or in thermal conditions. Habitats with higher food availability harboured more hatchlings, and feeding rates (obtained by stomach-flushing of recently-captured crocodiles) were highest in such areas. Predation risk may also differ among habitats: we were twice as likely to capture a crocodile after seeing it in open-bank sites than in the other two habitat types. Thus, habitat selection of hatchling crocodiles in this system may be driven both by prey availability and by predation risk.     

嗜水气单胞菌菌蜕的制备及其对银鲫的口服免疫

菌蜕系统是一个自身具有佐剂性质的新型疫苗体系,不含细胞质内容物但具有细菌的完整表面抗原结构,可诱导机体的体液、细胞免疫应答及增强黏膜免疫反应.本研究通过将带有裂解基因E的质粒pElysis转化至嗜水气单胞菌J-1株中,对Ah J-1(pElysis)进行温度诱导,温度从28℃升至42℃,每隔15min检测菌液的OD600值,测定其溶菌动力学,并做无菌检验,用扫描电镜观察裂解后的细菌形态,研究其作为口服疫苗对银鲫的效果.结果显示,通过温度诱导,嗜水气单胞菌J-1(pElysis)OD值在诱导30min后开始持续下降,75min时开始趋于平稳,到120min溶菌效率达99.99%,诱导16h后进行无菌检验,证实其无活菌.扫描电镜观察绝大部分菌体经诱导后形成菌蜕,细胞两端有溶菌通道.动物试验表明,用菌蜕口服免疫的银鲫,在第5周产生较高的凝集抗体,达到27,并能维持2周;而甲醛灭活苗组为26,维持时间仅一周;生理盐水对照组效价仅2.攻击试验表明,菌蜕疫苗组和甲醛灭活疫苗组对嗜水气单胞菌强毒株J.1的攻击均有保护作用,其相对保护率分别为16/20(78.95%)和12/20(57.9%),显示菌蜕疫苗比普通灭活疫苗能更有效地激活机体的免疫保护.     

Hematologic and plasma biochemical reference intervals for Morelet's crocodiles (Crocodylus moreletii) in the northern wetlands of Campeche, Mexico

Health surveys and hematologic and plasma biochemical analyses were conducted in 52 free-ranging and 51 captive Morelet's crocodiles (Crocodylus moreletii) in Campeche, Mexico, March-September 2007. Blood samples from 92 crocodiles (45 free-ranging and 47 captive) were collected for hematologic and plasma biochemical analyses. Average values of erythrocytes of free-ranging crocodiles were 1,046,166 cells/μl, and total white cells were 1.03 × 10(4) cells/μl. Captive crocodiles had erythrocyte and leukocyte values of 1,100,416 cells/μl and 8.51 × 10(3) cells/μl, respectively. There were no significant differences in values of erythrocytes or in hematocrit between free-ranging and captive crocodiles, or between sexes, or among size classes. Counts of leukocytes in free-ranging crocodiles were significantly higher than in captive individuals. The mean values of plasma analytes were 69.55 mg/l (glucose), 250.14 mg/l (cholesterol), 3.04 mg/l (uric acid), 2.70 mg/l (creatinine), and 20.20 IU/l (alanine aminotransferase). There were significant differences in cholesterol between free-ranging and captive crocodiles and between sexes.     

广东罗坑自然保护区鳄蜥生境选择的季节性差异

2006年3-10月,在广东省罗坑鳄蜥自然保护区利用直接观察法对鳄蜥春、夏、秋3季的生境选择进行研究。春、夏、秋3季各测定了31、71、31个有鳄蜥样方以及50、90、51个对照样方的14种生态因子。利用生态因子对比分析和逐步判别分析确定各季节生境选择的主要影响因素。结果表明,春季影响鳄蜥生境选择的主要影响因素是溪沟底质中沙的含量和植被盖度,正确判别率为97.5%;夏季影响鳄蜥生境选择的主要影响因素是溪沟底质中沙的含量、植被盖度、溪流宽度和枯枝百分比,正确判别率为94.4%;秋季影响鳄蜥生境选择的主要影响因素是溪沟底质中沙的含量、植被盖度和可利用食物量,正确判别率为97.6%。在区分春、夏、秋3季鳄蜥生境选择方面有一系列生态因子发挥作用,依照贡献值的大小依次为可利用食物量、干扰距离、枯枝百分比、溪水流速、溪沟坡度和溪宽等6个因子,由这6个变量构成的方程对春、夏、秋3季鳄蜥生境的正确区分率达到76.7%。判别函数的分析表明,春、夏、秋3季鳄蜥的生境选择具有较高的差异性,以判别函数建立的判别分类图表明,春、夏季以及夏、秋季鳄蜥的生境选择重叠较多,而秋季与春季的重叠较少。     

Environmental factors shape cloacal bacterial assemblages in great tit Parus major and blue tit P. caeruleus nestlings

Despite their potential ecological and evolutionary importance, factors shaping the composition of bacterial communities in wild vertebrate populations remain poorly understood. The goal of this study was to examine the relative contributions of environmental factors and genetic factors (e.g. species and common origin) to the variation of cloacal bacterial assemblages in wild bird nestlings. We conducted a partial cross-fostering experiment with two passerine species, the great tit Parus major and the blue tit P. caeruleus, sharing similar habitats and breeding biology. Nestlings of the two species were exchanged four days after hatching and cloacal bacteria were sampled nine days later. The structure of cloacal bacterial communities was determined by Ribosomal Intergenic Spacer Analysis. Our results showed that each nestling displayed a unique bacterial community. Furthermore, nestlings raised in the same nest shared significantly similar bacterial communities. The similarity of bacterial community was higher among heterospecific siblings raised within the same nest than between biological siblings raised in separate nests. Effects of common origin between species could not be detected and, if present, were dominated by nest-based short-term environmental effects. Our results show that growth conditions within nests and individually based endogenous factors have significant effects on cloacal bacteria assemblages and could affect post-fledging condition.     

Prevalence of potentially pathogenic culturable bacteria on eggshells and in cloacae of female Pied Flycatchers in a temperate habitat in central Spain

ABSTRACT Bacteria grow on avian eggshells and thus can potentially cause diseases in developing embryos. Little is known about culturable bacteria colonizing avian eggshells in free‐living birds, with most studies restricted to poultry. Our objective was to examine the culturable bacterial array growing on eggshells during incubation that could negatively affect hatching success of Pied Flycatchers (Ficedula hypoleuca) in a temperate montane habitat in central Spain. Cloacal culturable bacteria of females were also analyzed because bacteria can be vertically transmitted from females to eggs. We used fecal samples as surrogates of cloacal samples due to the small size of sampled birds. We found that eggshells and female cloacae of Pied Flycatchers harbored 24 and 40 bacterial families and species, respectively, but only a few in each clutch and each cloaca. Rod‐shaped gram‐negative bacteria and bacteria in the family Pseudomonadaceae were the most common bacteria on eggshells during early and late incubation and in female cloacae. Although based on small sample sizes, we found that females with rod‐shaped gram‐negative bacteria in their cloacae laid eggs that also had these bacteria, providing possible evidence for vertical transmission. We found no evidence for vertical transmission of Pseudomonadaceae, suggesting a possible environmental source for these bacteria. The prevalence of bacterial morphological types and major taxonomical categories on eggshells did not vary from early to late stages of incubation, providing support for the hypothesis that incubation may have bacteriostatic effects on bacterial proliferation on eggshells. Despite being primary egg invaders in poultry, we detected no effects of culturable Pseudomonadaceae or Pseudomonas luteola on hatching success. Our study represents the first to examine the culturable bacteria growing on the eggshells of a wild bird in a temperate habitat and additional studies based on culture‐independent techniques are required to confirm our results.     

Natal origin and dispersal of problem saltwater crocodiles in the Darwin Harbor,Australia

Management programs that successfully recovered wild saltwater crocodile (Crocodylus porosus) populations in the Northern Territory of Australia did so with an expanding commitment to maintaining public safety. One aspect of the program is the ongoing removal of resident and immigrant crocodiles within Darwin Harbor (since 1979), the main urban center. We determined the likely sources of crocodiles caught as problem animals between 2015–2017 by comparing recently developed methods for population assignment. Depending on the assignment model used, we estimated that between 30% and 50% of crocodiles in Darwin Harbor originated from the Adelaide and Mary rivers, and the Kakadu region east of Darwin, and between 20% and 30% of crocodiles originated from the Finniss, Reynolds, and Daly rivers southwest of Darwin. Saltwater crocodiles occur at particularly high densities in these catchments. The remainder came from a mixture of different sources across the Northern Territory. The most common animals captured were immature (150–180 cm) males that have traveled 100–200 km. We did not identify any relationships between the distance from the inferred origin to Darwin Harbor and the size and sex of the crocodiles, or the year of capture. The targeted removal of crocodiles from specific sites such as Darwin Harbor, near where most people live, improves public safety in the highest risk areas, without compromising abundant source populations in most areas.     

Species-level diversification of African dwarf crocodiles (Genus Osteolaemus): A geographic and phylogenetic perspective

The taxonomy of the African dwarf crocodile (genus Osteolaemus) has been disputed since a novel morphotype was discovered in the early 20th Century. Because this poorly-known reptile is widely hunted throughout the forests of Central and West Africa, resolving the existence and extent of taxonomic units has important management and conservation implications. Lack of molecular data from individuals of known origin and historical disagreement on diagnostic morphological characters have hindered attempts to settle one of the most important taxonomic questions in the Crocodylia. In an effort to clarify the evolutionary relationships among dwarf crocodiles, we sequenced three mitochondrial and two nuclear genes using a large sample of dwarf crocodiles from known localities across major drainage basins of forested Africa. Concordant results from Bayesian, maximum likelihood, maximum parsimony and population aggregation analytical methods support a previously recognized division of the dwarf crocodile into a Congo Basin form (O. osborni) and a West African form (Osteolaemus tetraspis), but also reveal a third diagnosable lineage from West Africa warranting recognition as an separate taxonomic unit. Corrected genetic distances between geographic regions ranged from 0.2% to 0.6% in nuclear fragments and 10.0 to 16.2% in mitochondrial COI. Population aggregation, using fixed and alternate character (nucleotide) states to cluster or divide populations, recovered 232 such molecular characters in 4286 bp of sequence data and unambiguously aggregated populations into their respective geographic clade. Several previously recognized morphological differences coincide with our molecular analysis to distinguish Congo Basin crocodiles from the Ogooué Basin and West Africa. Discrete morphological characters have not yet been documented between the latter two regions, suggesting further work is needed or molecular data may be required to recognize taxonomic divisions in cases where putative species are morphologically cryptic. This study highlights the importance of using widespread taxon sampling and a multiple evidence approach to diagnose species boundaries and reveal cryptic diversity.