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Isolation of Salmonella strains from reptile faeces and comparison of different culture media.

AIMS: To provide information on epidemiology and isolation of Salmonella strains from reptiles. METHODS AND RESULTS: Ninety-one samples collected from reptiles of the zoo of Rome or belonging to private owners were analysed using a standard protocol for isolation of Salmonella from food. Salmonella strains were tested for susceptibility to 15 antimicrobics by a disc-agar diffusion method. Forty-six samples (50.5%) were positive for Salmonella. Of the 22 strains serotyped, 17 belonged to Salmonella enterica subsp. I, four to the subsp. IIIa and one strain resulted untypeable. Rappaport-Vassiliadis broth (RVB) allowed to recover more Salmonella strains when bacterial growth in buffered peptone water (BPW) was scarce, while selenite cystine broth (SCB) was more efficient, whereas growth in BPW was abundant. The maximum isolation score was obtained by plating onto xylose lysine desoxycholate agar (XLD). The strains exhibited resistance at high percentages to colistin sulphate (58.7%), sulphamethoxazole (55.5%), streptomycin (32.6%), tetracycline (19.6%), ampicillin (17.4%) and nalidixic acid (13.1%). CONCLUSIONS: A high prevalence of Salmonella in reptiles was observed. For isolation, the choice of the enrichment broth depending on the degree of growth in BPW followed by plating onto XLD may be suggested. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper provides epidemiological data on the prevalence of Salmonella and laboratory protocols useful for isolation of Salmonella from faeces of reptiles.

Animals↗

Mammalian ZFY sequences exist in reptiles regardless of sex-determining mechanism.

In some reptiles, egg incubation temperature determines whether the embryo hatches as male or female; in others, sex chromosomes determine sex. A cloned gene (ZFY) representing the putative testis-determining factor in mammals was hybridized to genomic DNA of reptiles with sex chromosomes and to DNA of reptiles with temperature-dependent sex determination. No sex differences in hybridization patterns were observed. Hybridization of ZFY to polyadenylated RNA indicates that reptilian versions of this gene are expressed in embryos of both sexes during the temperature-sensitive period. If these highly conserved sequences are important in reptilian sex determination, then temperature-dependent and genotypic sex determination may have a similar molecular basis. For reptiles with XX/XY or ZZ/ZW systems, the absence of sex differences in hybridization patterns raises the question of whether the ZFY sequences reside on their sex chromosomes.

Animals↗

Genetic divergence of Campylobacter fetus strains of mammal and reptile origins.

Campylobacter fetus is a gram-negative bacterial pathogen of both humans and animals. Two subspecies have been identified, Campylobacter fetus subsp. fetus and Campylobacter fetus subsp. venerealis, and there are two serotypes, A and B. To further investigate the genetic diversity among C. fetus strains of different origins, subspecies, and serotypes, we performed multiple genetic analyses by utilizing random amplification of polymorphic DNA (RAPD), pulsed-field gel electrophoresis (PFGE), and DNA-DNA hybridization. All 10 primers used for the RAPD analyses can distinguish C. fetus strains of reptile and mammal origin, five can differentiate between C. fetus subsp. fetus and C. fetus subsp. venerealis strains, and four showed differences between type A and type B isolates from mammals. PFGE with SmaI and SalI digestion showed varied genome patterns among different C. fetus strains, but for mammalian C. fetus isolates, genome size was well conserved (mean, 1.52 +/- 0.06 Mb for SmaI and 1.52 +/- 0.05 Mb for SalI). DNA-DNA hybridization demonstrated substantial genomic-homology differences between strains of mammal and reptile origin. In total, these data suggest that C. fetus subsp fetus strains of reptile and mammal origin have genetic divergence more extensive than that between the two subspecies and that between the type A and type B strains. Combining these studies with sequence data, we conclude that there has been substantial genetic divergence between Campylobacter fetus of reptile and mammal origin. Diagnostic tools have been developed to differentiate among C. fetus isolates for taxonomic and epidemiologic uses.

Animals↗

Age estimation and longevity in reptiles.

After a brief recall of the classical meaning of the concept of longevity, the first part of this chapter describes and summarises the main current technique for the estimation of age in reptiles. Among them, sclerochronology is primarily taken into account. The cautious analysis of seasonal growth cycles recorded in hard tissues, although not as rigorous as the mark-release-recapture method of animals in their natural conditions, now appears as a rapid and reliable chronological tool already successfully used in individuals of many reptile species. Sclerochronology is especially efficient for the comparison of several populations, and it is the only method for fossils. The second part presents a synthetic review of known longevities and records in the different groups of reptiles. A short discussion about the significance of longevity shows that for reptiles, because of their thermic metabolism (ectothermy), the physiological longevity must be strongly distinguished from the chronological longevity, especially for a comparison with that known for birds and most mammals.

Aging↗

Growth in reptiles.

This paper reviews the literature on growth in reptiles. The current consensus is that growth data for small and medium-sized reptiles are best fitted by logistic-by-mass equations, data for larger reptiles by the von Bertalanffy equation. Experiments on the relationships between behavioural thermoregulation, food consumption and growth in both the laboratory and field are described, and an account is given of some recent work on individual differences in growth rates and their adaptive significance. In general, it may be said that in almost all areas of biology relating to growth, less is known about reptiles than about equivalent phenomena in fishes.

Animals↗

Ageing in reptiles.

Like in fishes, the reptiles appear to show three types of senescence. The African skink, Mabuya buettneri, shows rapid senescence similar to death at mating observed in Salmon and marsupial mouse. Most of the lizards and snakes undergo gradual senescence comparable to the pattern exhibited by a majority of vertebrates. On the other hand, turtles, tortoises and crocodiles continue to grow throughout life and are thus credited with slow or negligible senescence. Evidences and mechanisms of rapid or negligible senescence in reptiles are still fragmentary and unclear. Findings in a few species of lizards (Calotes versicolor) and snakes (Natrix natrix) showing gradual senescence support the concept of commonalities in ageing phenomena in vertebrates. An age-related increase in the stability of collagen and accumulation of altered enzyme molecules, a decrease in metabolism and response to stress-enhanced anti-oxidative defence mechanisms and the nature of responses to hormones, restricted diet and lower environmental temperature corroborate the concept. On the other hand neither the increase in mortality rate and accumulation of lipofuscin nor the reproductive senility have been shown conclusively in ageing reptile populations. It is likely that there are multiple mechanisms of senescence in reptiles. Further studies on selected species from among the 6,000 living species are necessary to unravel the phenomena.

Aging↗

Rehabilitation of native reptiles and amphibians in DuPage County, Illinois.

Between 1980 and 1994, 586 individuals of 20 native reptile and amphibian species were evaluated at a wildlife rehabilitation facility in DuPage County, Illinois (USA). On examination, 48% of reptiles and 52% of amphibians were injured, and 2% of each class were diseased; remaining animals were uninjured. The most frequent causes of injury to reptiles were automobile collisions (55%), ingestion of fishing tackle (19%), and domestic animal attacks (8%). Automobile collisions (38%), lawn and garden accidents (33%), and pet collection (19%) were the most frequent causes of injury to amphibians. We released 354 reptiles and 54 amphibians into appropriate habitat available in local forest preserves, representing case release prevalences of 84% and 68% for each class, respectively.

Amphibians↗

Reptile-associated salmonellosis--selected states, 1996-1998.

During 1996-1998, CDC received reports from approximately 16 state health departments of Salmonella infections in persons who had direct or indirect contact with reptiles (i.e., lizards, snakes, or turtles). Salmonella infection can result in invasive illness including sepsis and meningitis, particularly in infants. Despite educational efforts, some reptile owners remain unaware that reptiles place them and their children at risk for salmonellosis. This report summarizes clinical and epidemiologic information in four cases and provides information about state regulations to prevent transmission of Salmonella spp. from reptiles to humans.

Animals↗

Salmonellosis and arizonosis in the reptile collection at the National Zoological Park.

The prevalence of Salmonella and Arizona organisms in the reptile collection at the National Zoological Park was investigated. Culture of specimens from 311 reptiles, while live or at necropsy, yielded yielded 117 positive results, for an overall infection rate of 37%. Snakes had the highest rate, 55% (69 of 125); lizards had an intermediate rate, 36% (46 of 129); and turtles and tortoises had the lowest rate, 3% (2 of 63). Twenty-four serotypes of Salmonella enteritidis, 1 of S choleraesuis, and 39 of Arizona hinshawii were represented. While clinical illness was never directly attributed to infection with these organisms, pure cultures of Salmonella and Arizona were recovered at necropsy from some reptiles with gross and/or histologic lesions in the gastrointestinal tract, liver, spleen, and blood vessels. However, numerous other concurrent diseases and management problems were often considered the immediate cause of death, with Salmonella and Arizona being ready and significant opportunistic pathogens contributing to the demise of the reptiles.

Animals↗

Exercise performance of reptiles.

From the vantage point of thirty years of study, we can sketch the general features of activity capacity and performance ability in reptiles. Extant reptilian groups all share low levels of maintenance metabolism and ectothermy, with their consequent advantages (Pough, 1980) and disadvantages. Among the latter is a limited capacity to expand aerobic metabolism, limited in comparison to the relatively great costs of terrestrial locomotion. Particularly at low body temperatures, reptiles outstrip their aerobic capacities with any exercise more intense than a slow walk. Anaerobic metabolism, particularly anaerobic glycolysis, can be used to fuel bursts of intense activity. As a consequence, however, physiological disruption and exhaustion are entailed. Under field conditions, many reptiles alternate long periods of quiescence or slow movement with very brief bursts of exertion. Other ectotherms with a similar pattern of metabolism have been shown thereby to extend performance beyond that supportable by either aerobic or anaerobic metabolism alone (Weinstein and Full, 1992). Even with careful alternation between these metabolic modes, reptiles remain particularly prone to exhaustion during vigorous activity, as least as judged by our mammalian frame of reference. Their capacities for burst activity and exertion have been shown, at least in some species, to be important determinants of their natural survival.

Anaerobiosis↗

Estivation in South American amphibians and reptiles.

A number of amphibians and reptiles have cyclic behavior, becoming inactive with the coming of the dry season. In South America this pattern of activity is common, particularly in savannah-like vegetation. During the dry season amphibians burrow into the mud or soil, and either form a cocoon or increase the osmotic concentration of body fluids to reduce evaporative water loss. Some phyllomedusid tree frogs coat their body surface with skin secretion and excrete uric acid to minimize water loss. Reptiles also retreat into shelter deep enough to avoid temperature fluctuation during estivation or reduce metabolic response to temperature. Reduction of temperature sensitivity of the metabolism seems to be a strategy common to estivating amphibians and reptiles. Despite seasonal change of the environment, some species of reptiles are active all year round.

Amphibians↗

Experimental transmission of Cryptosporidium oocyst isolates from mammals, birds and reptiles to captive snakes.

Groups of four to five, 3-month-old rat snakes (Elaphe obsoleta) were separately gastrically inoculated with 2.0 x 10(6) viable oocysts of Cryptosporidium muris (mice and calves), C. muris-like (Bactrian camels), C. wrairi (guinea pigs), C. baileyi (chickens), C. meleagridis (turkeys), Cryptosporidium sp. (turtles, tortoises, chameleons and lizards) and C. serpentis from clinically (fatal case) and subclinically infected snakes. None of the snakes inoculated with oocysts originating from homothermous vertebrates developed infection as determined by histology and serology, whereas all snakes challenged with reptilian oocyst isolates were infected with Cryptosporidium on weeks 6 and 10 post-inoculation (PI). One week 10 PI, the snakes displayed mild to serve, multifocal to widespread, thinning and disorganization of gastric epithelium and nine out of twelve snakes infected by oocysts originating from reptiles other than snakes displayed severe gastric hyperplasia. Three out of ten snakes infected oocysts originating from snakes had ELISA-detectable Cryptosporidium-specific antibody (Ab) titers on week 6 PI; all snakes were Cryptosporidium-seroconverted on week 10 PI and their serum Ab titer significantly increased. The study demonstrated that Cryptosporidium infections in snakes maintained on the diet of rodents or birds cannot be initiated via ingestion of an infected food item; however, snakes can void ingested oocysts. Lack of host specificity among reptiles to this pathogen, demonstrated for the first time in the present study, indicates that snake-attributed C. serpentis is not distinct from Cryptosporidium sp. infecting reptiles other than snakes, and that clinical manifestations and virulence of Cryptosporidium in snakes in modulated by the species of the host. Housing of snakes with other reptiles can enhance transmission of Cryptosporidium to snakes, and therefore should be avoided.

Animals↗

The role of nitric oxide in regulation of the cardiovascular system in reptiles.

The roles that nitric oxide (NO) plays in the cardiovascular system of reptiles are reviewed, with particular emphasis on its effects on central vascular blood flows in the systemic and pulmonary circulations. New data is presented that describes the effects on hemodynamic variables in varanid lizards of exogenously administered NO via the nitric oxide donor sodium nitroprusside (SNP) and inhibition of nitric oxide synthase (NOS) by l-nitroarginine methyl ester (l-NAME). Furthermore, preliminary data on the effects of SNP on hemodynamic variables in the tegu lizard are presented. The findings are compared with previously published data from our laboratory on three other species of reptiles: pythons (), rattlesnakes () and turtles (). These five species of reptiles possess different combinations of division of the heart and structural complexity of the lungs. Comparison of their responses to NO donors and NOS inhibitors may reveal whether the potential contribution of NO to vascular tone correlates with pulmonary complexity and/or with blood pressure. All existing studies on reptiles have clearly established a potential role for NO in regulating vascular tone in the systemic circulation and NO may be important for maintaining basal systemic vascular tone in varanid lizards, pythons and turtles, through a continuous release of NO. In contrast, the pulmonary circulation is less responsive to NO donors or NOS inhibitors, and it was only in pythons and varanid lizards that the lungs responded to SNP. Both species have a functionally separated heart, so it is possible that NO may exert a larger role in species with low pulmonary blood pressures, irrespective of lung complexity.

Animals↗

Purification, characterization and comparison of reptile lysozymes.

Cation exchange column chromatography and gel filtration chromatography were used to purify four reptile lysozymes from egg white: SSTL A and SSTL B from soft shelled turtle (Trionyx sinensis), ASTL from Asiatic soft shelled turtle (Amyda cartilagenea) and GSTL from green sea turtle (Chelonia mydas). The molecular masses of the purified reptile lysozymes were estimated to be 14 kDa by SDS-PAGE. Enzyme activity of the four lysozymes could be confirmed by gel zymograms and showed charge differences on native-PAGE. SSTL A, SSTL B and ASTL had sharp pH optima of about pH 6.0, which contrasts with that of GSTL, which showed dual pH optima at about pH 6.0 and pH 8.0. The activities of the reptile lysozymes rapidly decreased within 30 min of incubation at 90 degrees C except for ASTL, which was more stable. Partial N-terminal amino acid sequencing and peptide mapping strongly suggested that the enzymes were C-type lysozymes. Interestingly, the mature SSTL lysozymes show an extra Gly residue at the N-terminus, which was previously found in soft-shelled turtle lysozyme. The reptile lysozymes showed lytic activity against several species of bacteria, such as Micrococcus luteus and Vibrio cholerae, but showed only weak activity to Pseudomonas aeruginosa and lacked activity towards Aeromonas hydrophila.

Amino Acid Sequence↗

Nondestructive indices of trace element exposure in squamate reptiles.

Compared with birds, mammals, fish, and even amphibians, very little is known about the effects of contaminants on reptiles. Recent evidence that many reptile populations may be declining has stimulated demand for toxicological studies of reptiles as well as development of nondestructive sampling techniques useful for assessing and monitoring contaminant exposure. The current study experimentally evaluated the utility of shed skins, tail clips, and blood samples as nondestructive indices of trace element exposure in banded water snakes, Nerodia fasciata. For 13.5 months, snakes were either fed fish from a coal ash-contaminated site or uncontaminated food from a reference site. Snakes fed contaminated prey accumulated As, Cd, Se, Sr, and V in various organs (i.e. liver, kidney, and/or gonads). Moreover, non-parametric discriminant function analysis revealed that snakes could be placed in two groups that reliably reflected their experimental diet based upon Se, Sr, and As concentrations in tail clips, blood, and/or shed skins. We suggest that nondestructive sampling techniques, particularly analyses of blood and tail clips, may be easily applied in evaluations of contaminant exposure in the field and laboratory and may prevent excessive destructive sampling of potentially threatened reptile species.

Animals↗

Exotic reptile bites.

Reptiles are a growing part of the exotic pet trade, and reptile bites have been considered innocuous in the emergency medicine literature. Two cases are reported of reptile bites, one from a green iguana and the other from a reticulated python. The treatment concerns associated with reptile bites are discussed.

Adult↗

Modeling reptile virus infection in vitro using Python regius airway organoids.

Zoonoses pose substantial global health risks, highlighting the need to better understand animal-to-human transmission. Reptiles are increasingly recognized as hosts of diverse pathogens, including numerous viruses, yet the diversity and prevalence of reptile pathogens, as well as their potential risk to humans, remain poorly understood. Here, we establish and characterize airway organoids derived from Python regius, providing an in vitro model to study reptile airway infection. Through de novo assembly of a Python regius reference genome, we characterize airway organoids at single-cell resolution, which suggests the presence of diverse cell populations including ionocytes, ciliated, secretory, goblet, endocrine, tuft, and basal cells. The organoids support productive infection with Ball Python Nidovirus (BPNV) and mount a robust epithelial antiviral response through the induction of interferon-stimulated genes, cytokines, and genes involved in chemical defense. As a proof-of-concept, treating organoids with antiviral drugs during infection reduces BPNV levels, highlighting the model's utility for drug testing. By providing a reductionist system of the serpentes airway, these organoids constitute a physiologically relevant in vitro model to study reptile viruses and host-pathogen interactions in their native host.

Animals↗

Environmental sex determination in a reptile varies seasonally and with yolk hormones.

Most hypotheses that have been put forward in order to explain the persistence of environmental sex determination (ESD) in reptiles assume a relatively fixed association of sex with temperature-induced phenotype and no maternal influence on offspring sex. Here we demonstrate the association of maternally derived yolk hormone levels with the offspring sex ratio and describe two new aspects of temperature-dependent sex determination (TSD), i.e. seasonal variation in both thermal response and yolk steroid levels. Eggs from painted turtles (Chrysemys picta) were incubated at 28 degrees C. The hatchling sex ratio at 28 degrees C (i.e. the phenotypic reaction norm for sex at 28 degrees C) shifted seasonally from ca. 72% male to ca. 76% female. Yolk oestradiol (E2) increased seasonally while testosterone (T) decreased. The proportion of males in a clutch decreased as E2 levels increased and the E2:T ratio increased. These new findings are discussed in relation to heritability and adaptive explanations for the persistence of ESD in reptiles. Maternally derived yolk hormones may provide a mechanism for the seasonal shift in the sex ratio which in turn may help explain the persistence of ESD in reptiles. They may also explain those clutches of other reptiles with TSD that fail to yield only males at maximally masculinizing conditions.

Animals↗