Saurian-associated Leishmania tarentolae in dogs: Infectivity and immunogenicity evaluation in the canine model.

In canine leishmaniosis endemic areas, Leishmania infantum may occur in sympatry with the non-pathogenic Leishmania tarentolae, which is associated to reptiles. The potential infectivity of L. tarentolae for mammals raises questions about the interactions between the two Leishmania species, and the potential cross-immune protection in dogs. This study aimed to assess the outcome of experimental L. tarentolae infection in dogs, determining: i) the anti-L. tarentolae antibody production, ii) the duration of the immunity and cytokine expression, and iii) the possible pathogenic effect in the canine host. Twelve purpose-bred beagle dogs were randomly allocated to three groups (intravenous inoculation, G1; intradermal inoculation, G2; negative control, G3). G1 and G2 dogs were inoculated twice (day 0, day 28) with 108 promastigotes of L. tarentolae strain (RTAR/IT/21/RI-325) isolated from a Tarentola mauritanica gecko. The animals were followed until day 206. Blood, serum, conjunctival swabs and lymph node aspirate samples were collected monthly and bone marrow, liver and spleen biopsies on day 91. Hematological and biochemical parameters were assessed monthly, as well as serology (IFAT and ELISA) and molecular identification of L. tarentolae. Mononuclear cells (PBMC) were obtained to assess the cytokine expression through in vitro stimulation or (re-) infection. Data from this study demonstrated that DNA from L. tarentolae is detectable up to 3 months post-infection, with seroconversion after day 28. Moreover, the non-pathogenic nature of L. tarentolae was confirmed, with a neutral Th1/Th2 polarization, and a possible shift to Th1 phenotype after derived macrophages (re-) infection, as demonstrated by the expression of IFN-gamma. Therefore, L. tarentolae demonstrated a great potential as a surrogate pathogen and/or immune-prophylaxis/immune-therapy against Leishmania infections in dogs and humans.

Corticosterone treatment results in fat deposition and body mass maintenance without effects on feeding behaviour or immunity in female lizards (Tropidurus catalanensis).

Throughout life, animals must maintain homeostasis while coping with challenging events. The period after reproduction can be challenging for oviparous females to maintain homeostasis since they direct most of their energy stores to vitellogenesis, possibly increasing the vulnerability to stressors. Changes in glucocorticoids' (GC) secretion promote various behavioural and physiological adjustments daily and to restore balance after facing stressors. However, when GC are elevated for extended periods, which usually occurs in response to chronic exposure to stressors, they can affect feeding behaviour and suppress the immune function. We aim to elucidate the effects of chronic corticosterone (CORT) exposure on feeding behaviour, body condition and immune function in female lizards, Tropidurus catalanensis, in the post-reproductive period. Thirty animals were divided into three groups: 1. Control (no experimental procedure performed); 2. Empty Implant (animals implanted with empty silastic tube); and 3. CORT Implant (animals implanted with silastic tube filled with CORT, with a chronic continuous release for at least a week). CORT plasma levels feeding behaviour, body condition (body index [BI] and fat index [FI]), leukocyte count, and several immune function variables (bacterial killing ability [BKA], hemagglutination titer, phytohemagglutinin [PHA] immune challenge and leukocyte count) were evaluated. After implantation, CORT treated animals maintained stable body mass through the experiment, while Control and Empty Implant groups displayed weight loss. In the CORT treated animals, there was also a positive relation between BI and FI, and higher FI when compared to groups 1 and 2. No effects of CORT were observed on feeding behaviour nor on the immune function.

Transcriptomic profiling revealed immune-related signaling pathways in response to experimental infection of Leishmania donovani in two desert lizards from Northwest China.

Little is known about the immune response of lizards to Leishmania parasties. In this study, we conducted the first liver transcriptome analysis of two lizards (Phrynocephalus przewalskii and Eremias multiocellata) challenged with L. donovani, endemic to the steppe desert region of northwestern China. Our results revealed that multiple biological processes and immune-related signaling pathways are closely associated with the immune response to experimental L. donovani infection in the two lizards, and that both lizards show similar changes to mammals in terms of immunity to Leishmania. However, the interspecific divergence of the two lizards leads to different transcriptomic changes. In particular, in contrast to P. przewalskii, the challenged E. mutltiocellata was characterized by the induction of down-regulation of most DEGs. These findings will contribute to the scarce resources on lizard immunity and provide a reference for further research on immune mechanisms in reptiles.

Heterologous Expression of Human IFNγ and Anti-IL17 Antibody in Leishmania tarentolae Promastigote.

BACKGROUND: Leishmania is an intracellular flagellate protozoan parasite that causes a wide range of clinical diseases in humans. The basis of immunological resistance against leishmaniasis depends on Thl reactions and is within the time period of cytokine function. METHODS: In this study, human anti-IL17 antibody and IFNγ-producing promastigote were produced to be used in leishmanization. A sequence of light and heavy chains' gene of anti-IL17 antibody and human IFNγ (hIFNγ) was obtained from the NCBI database and synthesized in the ECORV reaction site in the plasmid pGH, which it's called pGH-hIFNγ-antiIL17. The synthesized part using the restriction enzyme ECORV was extracted from the plasmid and after purification by electroporation was transferred to Iranian lizard Leishmania (I.L.L). Evaluation of structural presence in the I.L.L genome at the level of DNA and mRNA was assessed. The expressions of hIFNγ and anti-IL17 were evaluated and confirmed using ELISA and western blot analysis. The hIFNγ secreted from the culture medium was collected at high concentrations of 124.36 ± 6.47 pg/mL. RESULTS: Targeted gene replacement into the I.L.L genome was successfully performed for the first time using the pGH-hIFNγ-antiIL17 plasmid in an identical replacement process. Stabilized recombinant DNA contains a target gene that has no toxicity to the parasite. CONCLUSIONS: The effective achievement of producing a recombinant gene was done for the first time by replacing the I.L.L-CPC gene with plasmid pGH-hIFNγ-antiIL17 by targeted gene replacement. This cab can regulate the production of hIFNγ and anti-IL17. This makes it a viable choice for eliminating leishmania.

Outer surface protein E (OspE) mediates Borrelia burgdorferi sensu stricto strain-specific complement evasion in the eastern fence lizard, Sceloporus undulatus.

In North America, Lyme disease is primarily caused by the spirochetal bacterium Borrelia burgdorferi sensu stricto (Bb), which is transmitted between multiple vertebrate hosts and ixodid ticks, and is a model commonly used to study host-pathogen interactions. While Bb is consistently observed in its mammalian and avian reservoirs, the bacterium is rarely isolated from North American reptiles. Two closely related lizard species, the eastern fence lizard (Sceloporus undulatus) and the western fence lizard (Sceloporus occidentalis), are examples of reptiles parasitized by Ixodes ticks. Vertebrates are known to generate complement as an innate defense mechanism, which can be activated before Bb disseminate to distal tissues. Complement from western fence lizards has proven lethal against one Bb strain, implying the role of complement in making those lizards unable to serve as hosts to Bb. However, Bb DNA is occasionally identified in distal tissues of field-collected eastern fence lizards, suggesting some Bb strains may overcome complement-mediated clearance in these lizards. These findings raise questions regarding the role of complement and its impact on Bb interactions with North American lizards. In this study, we found Bb seropositivity in a small population of wild-caught eastern fence lizards and observed Bb strain-specific survivability in lizard sera. We also found that a Bb outer surface protein, OspE, from Bb strains viable in sera, promotes lizard serum survivability and binds to a complement inhibitor, factor H, from eastern fence lizards. Our data thus identify bacterial and host determinants of eastern fence lizard complement evasion, providing insights into the role of complement influencing Bb interactions with North American lizards.

The Crystal Structure of the MHC Class I (MHC-I) Molecule in the Green Anole Lizard Demonstrates the Unique MHC-I System in Reptiles.

The reptile MHC class I (MCH-I) and MHC class II proteins are the key molecules in the immune system; however, their structure has not been investigated. The crystal structure of green anole lizard peptide-MHC-I-ß2m (pMHC-I or pAnca-UA*0101) was determined in the current study. Subsequently, the features of pAnca-UA*0101 were analyzed and compared with the characteristics of pMHC-I of four classes of vertebrates. The amino acid sequence identities between Anca-UA*0101 and MHC-I from other species are <50%; however, the differences between the species were reflected in the topological structure. Significant characteristics of pAnca-UA*0101 include a specific flip of ∼88° and an upward shift adjacent to the C terminus of the α1- and α2-helical regions, respectively. Additionally, the lizard MHC-I molecule has an insertion of 2 aa (VE) at positions 55 and 56. The pushing force from 55-56VE triggers the flip of the α1 helix. Mutagenesis experiments confirmed that the 55-56VE insertion in the α1 helix enhances the stability of pAnca-UA*0101. The peptide presentation profile and motif of pAnca-UA*0101 were confirmed. Based on these results, the proteins of three reptile lizard viruses were used for the screening and confirmation of the candidate epitopes. These data enhance our understanding of the systematic differences between five classes of vertebrates at the gene and protein levels, the formation of the pMHC-I complex, and the evolution of the MHC-I system.

Eurythermic Sprint and Immune Thermal Performance and Ecology of an Exotic Lizard at Its Northern Invasion Front.

AbstractThermal performance of immunity has been relatively understudied in ectotherms, especially in the context of invasive species or in relation to other fitness-related traits and thermoregulatory patterns in the field. For reptiles, thermal biology is a primary factor determining physiological performance and population viability, and suboptimal thermal conditions may limit the expansion of exotic species along the edges of their invasion fronts. This study examined thermoregulatory ecology and thermal performance of immunity and sprinting in a population of Mediterranean geckos (Hemidactylus turcicus) at the northern edge of their invasion front in a temperate zone of the United States. In the field, we quantified temperatures of geckos of varied age classes in relation to air, wall, and refugia temperatures. We also quantified temperature-dependent sprint performance and immune function in field-collected geckos to detail thermal performance patterns that may contribute to the capacity for this species to invade cool climates. Although body temperature (Tb) of wild-caught geckos correlated with wall temperature, average Tb exhibited wide distributions, suggesting eurythermy. Furthermore, the thermal performance of immune swelling responses to phytohemagglutinin injections and sprinting was optimized over a similarly wide temperature range that overlapped with the field Tb's that suggest eurythermy in this species. The wide thermal performance breadths in these traits could buffer against variation in factors such as pathogen exposure and environmental temperatures that could otherwise suppress functional performance. Thus, eurythermy of sprint and immune performance may facilitate the invasive potential of H. turcicus.

Ecoimmune reallocation in a native lizard in response to the presence of invasive, venomous fire ants in their shared environment.

Exposure to stressors over prolonged periods can have fitness-relevant consequences, including suppression of immune function. We tested for effects of presence of an invasive species threat on a broad panel of immune functions of a coexisting lizard. Eastern fence lizards (Sceloporus undulatus) have been exposed to invasive fire ants (Solenopsis invicta) for over 80 years. Fire ants sting and envenomate lizards, causing physiological stress, but we do not have a comprehensive understanding of the broad immune consequences of lizard exposure to fire ant presence. We conducted a suite of immune measures on fence lizards caught from areas with long histories of fire ant invasion and lizards from areas not yet invaded by fire ants. The effect of fire ant presence on immunity varied depending on the immune component measured: within fire ant invaded areas, some portions of immunity were suppressed (lymphocytic cell-mediated immunity, complement), some were unaffected (phagocytic respiratory burst, natural antibodies), and some were enhanced (anti-fire ant immunoglobulin M, basophils) compared to within uninvaded areas. Rather than fire ants being broadly immunosuppressing, as generally assumed, the immune response appears to be tailored to this specific stressor: the immune measures that were enhanced are important to the lizards' ability to handle envenomation, whereas those that were unaffected or suppressed are less critical to surviving fire ant encounters. Several immune measures were suppressed in reproductive females when actively producing follicles, which may make them more susceptible to immunosuppressive costs of stressors such as interactions with fire ants.

Decrease in preferred temperature in response to an immune challenge in lizards from cold environments in Patagonia, Argentina.

In ectotherms, the likelihood of surviving an infection is determined by the efficiency of thermoregulation, the availability of a variety of thermal microenvironments, the individual's health status, and the virulence of the infective agent. Physiological and behavioral demands related to an efficient immune response entail a series of costs that compete with other vital activities, specifically energy storage, growth, reproduction, and maintenance functions. Here, we characterize the thermal biology and health status by the presence of injuries, ectoparasites, body condition, and individual immune response capacity (using phytohemagglutinin in a skin-swelling assay) of the southernmost lizards of the world, Liolaemus sarmientoi, endemic to a sub-optimal, cold environment in Patagonia, Argentina. In particular, we study the effect of a bacterial endotoxin (lipopolysaccharide; LPS-treatment) on thermoregulation. We found that the field-active body temperature (Tb) was much lower than the preferred body temperature (Tp) obtained in the laboratory. All the individuals were in good body condition at the beginning of the experiments. The phytohemagglutinin test caused detectable thickening in sole-pads at 2 h and 24 h post-assay in males and non-pregnant females, indicating a significant innate immune response. In the experimental immune challenge, the individuals tended to prefer a low body temperature after LPS-treatment (2 h post-injection) and developed hypothermia, while the control individuals injected with phosphate buffered saline (PBS), maintained their body temperature throughout the trial. In both the LPS-treatment and PBS-control individuals, BC declined during the experiment. Hypothermia may allow this southernmost species to optimize the use of their energetic resources and reduce the costs of thermoregulation in a cold-temperate environment where they rarely attain the mean Tp (35.16 °C) obtained in laboratory.

Glucocorticoids, energy metabolites, and immunity vary across allostatic states for plateau side-blotched lizards (Uta stansburiana uniformis) residing in a heterogeneous thermal environment.

Reptiles rely on thermal heat exchange to achieve body temperatures (Tbody ) conducive to maintaining homeostasis. Diurnal changes in the thermal environment are therefore liable to influence allostatic mediation of survival processes (e.g., immunity) during environmental challenges or stressors. However, the extent to which Tbody prompts individual variation in physiology remains largely unexplored in reptiles. Our study tested how circulating energy-mobilizing hormone, energy metabolites, and immunity can vary across basal and stress-induced allostatic states for plateau side-blotched lizards (Uta stansburiana uniformis) residing in a heterogeneous thermal environment. We collected baseline and acute stress blood samples from male lizards to compare changes in plasma corticosterone (CORT), glucose, and bacterial killing ability (BKA) in relation to each other and Tbody . We hypothesized each physiological parameter differs between allostatic states, whereby stress-induced activity increases from baseline. At basal and stress-induced states, we also hypothesized circulating CORT, glucose, and BKA directly correspond with each other and Tbody . We found both CORT and BKA increased while glucose instead decreased from acute stress. At basal and stress-induced allostatic states, we found CORT to be directly related to Tbody while BKA was inversely related to CORT. We also found BKA and glucose were directly related at baseline, but inversely related following acute stress. Overall, these results demonstrate allostatic outcomes from acute stress in a free-living reptile and the role of temperature in mediating energetic state and immunity. Future research on reptilian allostasis should consider multiple environmental conditions and their implications for physiological performance and survival.

Decreased Cell-Mediated Immune Response in Bosk's Fringe-Toed Lizards (Acanthodactylus boskianus) Inhabiting an Industrialized Area in Southern Tunisia.

Lizards increasingly are recognized as suitable contaminant biomonitors in terrestrial ecosystems. Previously, we have shown that Bosk's fringe-toed lizards (Acanthodactylus boskianus) living close to the Gabès-Ghannouche industrial complex for fertilizer and acid production in southern Tunisia were contaminated by heavy metals. However, the impact of this contamination on lizard health parameters has not been investigated. In this study, we used the phytohaemagglutinin (PHA) skin-swelling test to assess whether the proximity to the industrial complex was associated with notable changes in lizard cell-mediated immune response (CMI). Our results showed significantly lower CMI in lizards living close to the industrial complex compared to those occurring farther away in a similar coastal habitat. Overall, our findings are consistent with the idea of immunotoxic effects of metal contamination. They also stress the usefulness of the PHA approach as an efficient tool for the evaluation of contaminant-related immunosuppression in lizards.

Experimental warming induces oxidative stress and immunosuppression in a viviparous lizard, Eremias multiocellata.

Reptiles are especially vulnerable to climate warming because their behavior, physiology, and life history are highly dependent on environmental temperature. In this study, we envisaged new probable mechanisms underlying the high vulnerability of lizards, wherein heat exposure induces oxidative stress and leads to immunosuppression. To test this hypothesis, we conducted a warming experiment on a lizard (Eremias multiocellata) from a desert steppe in Inner Mongolia from May to September using open-top chambers set up in their natural habitat and compared the components of oxidative stress (antioxidant ability [Superoxide dismutase (SOD) activity], extent of oxidative damage [malondialdehyde (MDA) content]), and immunocompetence (white blood cells [WBC] counts and immunoglobulin M [IgM] expression) between the warming and control groups. At the end of the experiment, the warming treatment did not affect the survival rate of the lizards. However, MDA content, but not SOD activity, was significantly higher in the warming group than in the control group. The WBC counts and IgM expression were significantly lower in the warming group than in the control group. Our results verified our hypothesis and provided novel cues and methods for the investigation of the mechanisms behind the high probability of extinction of other ectotherms under warming conditions.

Impact of immunological state on eco-physiological variables in one of the southernmost lizards in the world.

The immune state is an essential component of survival as it directly influences physiological performance and health status. Variation in the leukocyte profile, a significantly increase in body temperature, and a detriment of the eco-physiological performance are among the possible consequences of an unhealthy state. In this study we analyse and discuss how field body temperature, preferred body temperature, the speed for sprint and long runs, locomotor stamina, and body condition can be affected by the immunological state (i.e. leukocyte profile) in a wild population of Liolaemus sarmentoi. Juveniles and adult males with a high percentage of eosinophils, basophils, and a low percentage of monocytes preferred higher body temperatures in a thermal gradient, while pregnant females maintained thermal preferences independently of leukocyte profile. Although juveniles with a high percentage of heterophils showed less locomotor stamina, adult males and pregnant females showed no differences in locomotor performance in relation to leukocyte profile. This study represents a starting point in eco-immunology of a wild lizard population of Liolaemus in cold and temperate environments of Patagonia where the southward shift in the geographic ranges of pathogen populations due to global warming represents a threat to resident host populations.

Investment of both essential fatty and amino acids to immunity varies depending on reproductive stage.

Trade-offs among the key life-history traits of reproduction and immunity have been widely documented. However, the currency in use is not well-understood. We investigated how reproducing female side-blotched lizards, Uta stansburiana, allocate lipids versus proteins when given an immune challenge. We tested whether lizards would invest more in reproduction or immunity depending on reproductive stage. Females were given stable isotopes (15 N-leucine and 13 C-1-palmitic acid), maintained on a regular diet and given either a cutaneous biopsy or a sham biopsy (control). Stable isotopes were monitored and analyzed in feces and uric acid, skin biopsies, eggs, and toe clips. We found that lizards deposited both proteins and lipids into their healing wounds (immune-challenged), skin (control), and eggs (all) and that catabolism of proteins exceeded incorporation into tissue during wound-healing. Specifically, we found that healed biopsies of wounded animals had more leucine and palmitic acid than the nonregrown skin biopsies taken from unwounded control animals. Earlier in reproduction, lizards invested relatively more labeled proteins into healing their wound tissue, but not into unwounded skin of control animals. Thus, reproduction is sometimes favored over self-maintenance, but only in later reproductive stages. Finally, we documented positive relationships among the amount of palmitic acid deposited in the eggs, the amount of food eaten, and the amount of palmitic acid excreted, suggesting higher turnover rates of lipids in lizards investing highly in their eggs.

The Komodo dragon (Varanus komodoensis) genome and identification of innate immunity genes and clusters.

BACKGROUND: We report the sequencing, assembly and analysis of the genome of the Komodo dragon (Varanus komodoensis), the largest extant lizard, with a focus on antimicrobial host-defense peptides. The Komodo dragon diet includes carrion, and a complex milieu of bacteria, including potentially pathogenic strains, has been detected in the saliva of wild dragons. They appear to be unaffected, suggesting that dragons have robust defenses against infection. While little information is available regarding the molecular biology of reptile immunity, it is believed that innate immunity, which employs antimicrobial host-defense peptides including defensins and cathelicidins, plays a more prominent role in reptile immunity than it does in mammals. . RESULTS: High molecular weight genomic DNA was extracted from Komodo dragon blood cells. Subsequent sequencing and assembly of the genome from the collected DNA yielded a genome size of 1.6 Gb with 45x coverage, and the identification of 17,213 predicted genes. Through further analyses of the genome, we identified genes and gene-clusters corresponding to antimicrobial host-defense peptide genes. Multiple ß-defensin-related gene clusters were identified, as well as a cluster of potential Komodo dragon ovodefensin genes located in close proximity to a cluster of Komodo dragon ß-defensin genes. In addition to these defensins, multiple cathelicidin-like genes were also identified in the genome. Overall, 66 ß-defensin genes, six ovodefensin genes and three cathelicidin genes were identified in the Komodo dragon genome. CONCLUSIONS: Genes with important roles in host-defense and innate immunity were identified in this newly sequenced Komodo dragon genome, suggesting that these organisms have a robust innate immune system. Specifically, multiple Komodo antimicrobial peptide genes were identified. Importantly, many of the antimicrobial peptide genes were found in gene clusters. We found that these innate immunity genes are conserved among reptiles, and the organization is similar to that seen in other avian and reptilian species. Having the genome of this important squamate will allow researchers to learn more about reptilian gene families and will be a valuable resource for researchers studying the evolution and biology of the endangered Komodo dragon.

Feeling the heat: Extreme temperatures compromise constitutive innate humoral immunity and skin color in a desert dwelling lizard.

Environmental temperature, particularly in habitats with extreme temperature fluctuations, may shape selection pressures on life history traits. Especially in ectotherms, temperature affects performance, physiology, and in some species, skin color. Skin color can be a sexual ornament signaling the bearer's ability to resist infections, when only high-quality individuals are able to invest both in high immune defense and elaborate ornament expression. However, how the information content of these sexual traits may vary with environmental conditions has been less studied. Dickerson's collared lizard (Crotaphytus dickersonae) males are blue and have a black and white collar. This conspicuous coloration signals performance and immune response, and is related to body temperature. Here, by maintaining males at higher, lower, and mean environmental temperatures we evaluated whether temperature variation influences color and constitutive innate humoral immunity (agglutination and lysis titers, estimated through hemolysis-hemagglutination assays), and whether extreme temperatures impose trade-offs between color and humoral immunity. We found that at low and high temperature treatments males had lower agglutination and lysis titers, and at low temperature, blue chroma from the dorsum declined and males became greener. Interestingly, at low and control temperature treatments, agglutination titer and blue coloration were positively correlated, whereas high temperatures revealed a trade-off between increasing agglutination titers and displaying bluer skin color. Our results suggest that in the Dickerson collared lizard even short-term variation of environmental temperature affects performance of constitutive innate humoral immunity and the brilliant blue skin color. Particularly, high temperatures may compromise some components of male's immunity and sexual signaling.

Population history with invasive predators predicts innate immune function response to early-life glucocorticoid exposure in lizards.

Early-life stress can suppress immune function, but it is unclear whether transgenerational stress exposure modulates the immune consequences of early stress. In populations where, historically, the immune system is frequently activated, e.g. persistent stressors that cause injury, it may be maladaptive to suppress immune function after early-life stress. Thus, the relationship between early-life stress and immune function may vary with population-level historical stressor exposure. We collected gravid fence lizards (Sceloporus undulatus) from populations that naturally differ in long-term exposure to invasive fire ants (Solenopsis invicta). We manipulated early-life stress in the resulting offspring via weekly exposure to fire ants, application of the stress-relevant hormone corticosterone or control treatment from 2 to 43 weeks of age. We quantified adult immune function in these offspring with baseline and antigen-induced hemagglutination and plasma bacterial killing ability. Early-life corticosterone exposure suppressed baseline hemagglutination in offspring of lizards from populations not exposed to fire ants but enhanced hemagglutination in those from populations that were exposed to fire ants. This enhancement may prepare lizards for high rates of wounding, toxin exposure and infection associated with fire ant attack. Adult bacterial killing ability and hemagglutination were not affected by early-life exposure to fire ants, but the latter was higher in offspring of lizards from invaded sites. A population's history of persistent stress may thus alter individual long-term immunological responses to early-life stressors. Further consideration of historical stressor exposure (type and duration) may be important to better understand how early-life stressors affect adult physiology.

Leptin ameliorates the immunity, but not reproduction, trade-off with endurance in lizards.

Life-history trade-offs result from allocation of limited energetic resources to particular traits at the expense of others. When resources are scarce, some traits will take priority over others in the degree of their expression. For example, the current reproduction may be sacrificed to enhance survival. Although intuitive from an evolutionary perspective, such priorities must be based on proximate mechanisms that respond to the current conditions. The hormone leptin serves as a signal of energy availability in vertebrates, and has been proposed as a mediator of energy allocation between reproduction and traits that enhance survival, such as the immune system. However, since leptin affects reproduction and immunity in a similar way, it remains unclear which takes priority when energy availability is low. Green anole lizards (Anolis carolinensis) with increased activity, via exercise training, have a marked decrease in immune function as well as reproduction, especially when calories are restricted. We hypothesized that endurance training and calorie restriction would lower immune and reproductive function due to energy limitation, and supplemental leptin would 'rescue' either immune function or reproduction (or both) due to the hormonal signal that energetic resources are available. We found that supplementary leptin rescued immune function in calorie-restricted, trained lizards, but reproduction was not rescued in males or females. This suggests that immune function and reproduction have different sensitivities to leptin in both sexes, or that reproduction is more energy limited and takes low priority even when the signal of energy availability is present.

Effects of cypermethrin (pyrethroid), glyphosate and chlorpyrifos (organophosphorus) on the endocrine and immune system of Salvator merianae (Argentine tegu).

Several geographical areas where Salvator merianae is distributed in Argentina are included in regions with agricultural activity and exposed to pesticide formulations. Some pesticides could affect defense mechanisms being able alter structures of some components of immune and endocrine systems. To assess the potential effects of pesticides in this reptile under seminatural conditions, on the immune system and endocrine responses in S. merianae we analyzed several blood parameters. Total (TWBCC), differential (DWBCC) white blood cells count, heterophils/lymphocytes index (H/L), lobularity index (LI), natural antibodies (NAbs) titres, complement system (CS), and corticosterone concentration were analyzed in animals exposed to a mixture of cypermethrin (25%), glyphosate (66.2%) and chlorpyrifos (48%) formulations. In addition, body size was considered in these analyzes. TWBCC and NAbs revealed lower values in organisms exposed to pesticides respect to a control indicating a possible immunosuppression effect. Besides, the LI showed a greater number of lobes in organism exposed demonstrating symptoms of chronic infection. In addition, we observed a reduced growth in these animals possibly related to a less energy investment in body mass to maintain an active defense against pesticides. Finally, we found high levels of plasma corticosterone in animals exposed to mix formulation that could demonstrate neuroendocrine axis activation. Other parameters like DWBCC, H/L index and activity of CS showed no differences in treated animals respect to control group, which could indicate low sensibility of these parameters to the concentration of pesticides used. Our results provide evidence of the toxic effects of pesticides on different immune system parameters, but also a trade-off among these parameters, corticosterone levels and growth. In this way, we can conclude that the formulated pesticides applied widely and constantly in the areas occupied by S. merianae, would be affecting its immune and endocrine systems and therefore its ability to defend against external agents. This kind of studies is of great interest to know the possible responses of wild species to anthropogenic disturbances such as pesticide contamination.

Selection, drift, and introgression shape MHC polymorphism in lizards.

The major histocompatibility complex (MHC) has long served as a model for the evolution of adaptive genetic diversity in wild populations. Pathogen-mediated selection is thought to be a main driver of MHC diversity, but it remains elusive to what degree selection shapes MHC diversity in complex biogeographical scenarios where other evolutionary processes (e.g. genetic drift and introgression) may also be acting. Here we focus on two closely related green lizard species, Lacerta trilineata and L. viridis, to address the evolutionary forces acting on MHC diversity in populations with different biogeographic structure. We characterized MHC class I exon 2 and exon 3, and neutral diversity (microsatellites), to study the relative importance of selection, drift, and introgression in shaping MHC diversity. As expected, positive selection was a significant force shaping the high diversity of MHC genes in both species. Moreover, introgression significantly increased MHC diversity in mainland populations, with a primary direction of gene flow from L. viridis to L. trilineata. Finally, we found significantly fewer MHC alleles in island populations, but maintained MHC sequence and functional diversity, suggesting that positive selection counteracted the effect of drift. Overall, our data support that different evolutionary processes govern MHC diversity in different biogeographical scenarios: positive selection occurs broadly while introgression acts in sympatry and drift when the population sizes decrease.