Ontogenetic and interelemental study of appendicular bones of Caiman latirostris Daudin, 1802 sheds light on osteohistological variability in crocodylians.

The osteohistology of vertebrates provides a reliable source to deduce biological information, particularly regarding growth and development. Although osteohistological studies in Neosuchia (Crocodyliformes, Mesoeucrocodylia) are relatively numerous, the number of species studied within the group is still small. Extant crocodilians are known to exhibit intraspecific variability linked to environmental conditions, habitat, feeding, and other intrapopulation factors. Here, we analyzed the osteohistology of the living South American Caiman latirostris throughout posthatching ontogeny. The histology of several appendicular bones of 13 different-sized captive and wild individuals were examined. Although some thin sections revealed the classic lamellar, parallel-fibered, or woven bone matrices, others showed a variation and a mix between the organization of the bone tissue. These histological differences are likely related to variability in the growth dynamics of caimans. In some bones of the juveniles studied, remnants of embryonic bone were observed. Osteohistological variation related to prevailing environmental conditions is documented. Furthermore, our results show ontogenetic variation in the type of bone tissues deposited throughout the development of C. latirostris. This study offers a broad framework for life history interpretations for C. latirostris and provides insight into the evolutionary history and ontogenetic growth of extinct crocodylian lineages.

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.

Immuno-endocrine alterations and oxidative stress induced by commercial formulations of pesticides in the tegu lizard (Salvator merianae).

Recent research has shown the risk of environmental contamination by pesticides in Argentina. Salvator merianae is considered as "sentinel species" for monitoring their effects. We intended to investigate growth, immunological and endocrine functions, and oxidative stress in S. merianae exposed to two commercial brands of the herbicide glyphosate: Roundup and Panzer Gold, as well as, to two insecticides: chlorpyrifos, and cypermethrin. Our results showed a higher increase in growth in animals exposed to Roundup and chlorpyrifos and a lower increase in those exposed to Panzer Gold. Higher total leukocyte and lymphocyte count values were observed in lizards exposed to Panzer Gold. The heterophils/lymphocytes ratio decreased in all treated animals. Regarding lobularity index, natural antibodies titers, and complement system activity values, lizards exposed to cypermethrin had higher values. Low corticosterone levels were observed in animals exposed to Roundup. Our results reveal different toxic effects of pesticides contributing to the knowledge of the pesticides' potential deleterious impact in the habitat of S. merianae.

Effects of glyphosate, 2,4-D, chlorantraniliprole, and imidacloprid formulations, separately and in mixtures in Caiman latirostris hatchlings.

The present study demonstrated the potential of glyphosate (GLY), 2,4-dichlorophenoxyacetic acid (2,4-D), imidacloprid (IMI) and chlorantraniliprole (CAP) separately and in mixtures to induce oxidative stress and DNA damage in Caiman latirostris hatchlings. Under controlled condition, an embryonic exposure to these pesticides was done at concentrations recommended for soybean crops. Treatments were: negative control, GLY, 2,4-D, IMI, CAP, mixture 1 (M1): GLY + 2,4-D, M2: IM I + CAP and M3: GLY + 2,4-D + IMI + CAP. At hatching, blood samples were taken for the evaluation of genotoxicity, oxidative damage to lipids and DNA, the enzymatic activity of catalase (CAT) and superoxide dismutase (SOD), and the expression level of their corresponding genes (catalase: cat and superoxide dismutase: sod). It has been shown that IMI, M2 and M3 induced a significant inhibition of CAT activity while no effect was observed on SOD. In turn, lipid peroxidation was significantly higher in individuals exposed to IMI, and to all the mixtures. Besides, genotoxicity and oxidative DNA damage were observed in all exposed groups. The results of mRNA expression showed no difference at transcription levels. In the same way, no alterations in growth parameters were recorded at hatching. Regarding to the mixtures, we observed a potentiating action of IMI on M3 in lipid peroxidation as well as independent action on oxidative DNA damage and genotoxicity parameters. Our results highlight the importance of investigating the effect of pesticides and their mixtures considering the potential consequences to caimans living in natural environments.

Blood biochemistry parameters of Broad snouted caiman, Caiman latirostris subjected to stress conditions, appliying micro-volume techniques.

Plasma biochemistry values represent a crucial and minimally invasive tool for evaluating nutritional and physiological condition of the animal. Biochemical data provides valuable information in those species exposed to environmental stressors. In this study, we aimed to apply for the first time micro-volume plasma tests to measured total plasma protein, albumin, and glucose in juvenile Caiman latirostris and to compare our results with other studies reporting crocodilian biochemistry values. We found that caimans from heat treatment were heavier, longer and had higher levels of glucose than those from the control group. Conversely, food restricted caimans had less growth, body condition and had 32.1% lower albumin concentrations than those from the control group. While no significant differences were observed in any parameters measured from water restricted caimans compared with control group. The values obtained for each of the biochemical determinations were lower than those previously reported for adult of C. latirostris, but similar finding for protein concentration compared with Alligator mississippiensis. Micro technique offers the advantage of increasing information and at the same time a greater physiological understanding of the crocodilians species.

How the exposure to environmentally relevant pesticide formulations affects the expression of stress response genes and its relation to oxidative damage and genotoxicity in Caiman latirostris.

This study aimed to analyze the molecular stress responses thought the expression levels of catalase (cat), superoxide dismutase (sod) and heat shock protein 70 (hsp70) genes, and how these relate with cellular stress response considering oxidative damage to lipids, DNA and genotoxicity in blood of Caiman latirostris hatchlings exposed to pesticide formulations under ex situ conditions. Treatments were: negative control (NC-tap water), glyphosate 2% (GLY), cypermethrin 0.12% (CYP), chlorpyrifos 0.8% (CPF), and their ternary mixture (Mx3). The concentrations and schedule of application were those recommended in soybean crops. Soil and water showed pesticides residues in all exposed groups. Results showed a statistically significant increase in the micronucleus frequency and DNA damage, with an important oxidation in all exposed groups. The expression level of cat gene was significantly higher in CYP while the expression of hsp70 was significantly lower in GLY, CYP and Mx3, compared to NC. Pesticides tested showed alterations in expression levels, growth parameters, DNA damage and base oxidation under realistic exposure conditions, and can threaten, in the long term, the health status of wild populations.

Effects of glyphosate, cypermethrin, and chlorpyrifos on hematological parameters of the tegu lizard (Salvator merianae) in different embryo stages.

Different studies have demonstrated effects of pesticides during embryo development in vertebrates and stage-dependent effects, but there is no information concerning this for Salvator merianae. We evaluated the effects of the herbicides Glyphosate Roundup (GLY-RU) and Glyphosate Panzer Gold (GLY-PZ); and the insecticides Chlorpyrifos (CPF) and Cypermethrin (CYP), and their complex mixtures, at different concentrations in hematological parameters of S. merianae embryos at two different development stages. The analyzed parameters were Total and Differential White Blood Cells Count, Heterophils/Lymphocytes index (H/L), Lobularity index, and Natural Antibodies (Nabs titres), as well as growth, embryo mortality and birth delay. Heterophils decreased in the intermediate concentrations tested of CYP and GLY-RU, in animals exposed at 33-days development. Lymphocytes increased in the intermediate concentration tested of GLY-RU, and the H/L index decreased in the maximum concentration tested of GLY-RU. NAbs titres increased in those animals exposed to the maximum CYP concentration tested. However, animals exposed at 3/5-days development showed no differences among treatments in most of the analyzed parameters, suggesting a stage-dependent response. Nevertheless, those animals exposed to GLY-PZ showed lower Nabs titres in relation to negative control. These results suggest effects on different hematological parameters related to the immune system of S. merianae, according to the used pesticide (herbicide or insecticide), its concentration and commercial formulation (GLY-RU or GLY-PZ), and the stages of development of the exposed animals. Our results reveal the importance of carrying out studies that evaluate the effects of permanent exposure of living beings and their environments to these toxics.

Genotoxicity induced by widely used pesticide binary mixtures on Caiman latirostris (broad-snouted caiman).

In this study, we investigated the effects of three binary mixtures of pesticide formulations commonly used in soybean crop: Cypermethrin Atanor® (25% -CYP), Chlorpyrifos Lorsban 48E® (48% -CPF) and Glyphosate Roundup® Full II (66.2% -GLY) on broad-snouted caiman (Caiman latirostris) hatchlings exposed by voluntary immersion under controlled condition. Genotoxicity was evaluated in peripheral blood erythrocytes through the micronucleus (MN) test and other nuclear abnormalities (NAs) and besides, growth of caiman was analyzed in each experimental group. The results showed that pesticide formulations tested, at concentrations similar to those recommended for application in the field, induced an increase in the frequency of micronucleus (FMN; p = 0.001) and Notched nuclei (NN; p = 0.010) in the mixture CYP + CPF, while an increase in the frequency of buds and NN was observed in the mixture of GLY + CYP (pbuds = 0.016 and pNN = 0.021), compared to the vehicle control (VC). On the contrary, a possible antagonistic action was observed between the components in the mixture GLY + CPF. Growth was not affected in any exposed groups (p > 0.05). There was a clutch effect in the frequency of binucleated erythrocytes (BiN; p = 0.011), total length (TL; p = 0.001) and snout-vent length (SVL; p = 0.031). Biomarkers used in this study are considered important predictive tools for the evaluation of xenobiotics. In this study, we demonstrated genotoxicity of pesticide mixtures under conditions that simulate the real situation of exposure suffered by caiman and other wild species in Argentina.

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.

Analysis of morphological variability and heritability in the head of the Argentine Black and White Tegu (Salvator merianae): undisturbed vs. disturbed environments.

The heterogeneity of biotic and abiotic factors influencing fitness produce selective pressures that promote local adaptation and divergence among different populations of the same species. In order for adaptations to be maintained through evolutionary time, heritable genetic variation controlling the expression of the morphological features under selection is necessary. Here we compare morphological shape variability and size of the cephalic region of Salvator merianae specimens from undisturbed environments to those of individuals from disturbed environments, and estimated heritability for shape and size using geometric morphometric and quantitative genetics tools. The results of these analyzes indicated that there are statistically significant differences in shape and size between populations from the two environments. Possibly, one of the main determinants of cephalic shape and size is adaptation to the characteristics of the environment and to the trophic niche. Individuals from disturbed environments have a cephalic region with less shape variation and also have a larger centroid size when compared to individuals from undisturbed environments. The high heritability values obtained for shape and size in dorsal view and right side view indicate that these phenotypic characters have a great capacity to respond to the selection pressures to which they are subjected. Data obtained here could be used as an important tool when establishing guidelines for plans for the sustainable use and conservation of S. merianae and other species living in disturbed areas.

Baseline values of immunologic parameters in the lizard Salvator merianae (Teiidae, Squamata).

The genus Salvator is widely distributed throughout South America. In Argentina, the species most abundant widely distributed is Salvator merianae. Particularly in Santa Fe province, the area occupied by populations of these lizards overlaps with areas where agriculture was extended. With the aim of established baseline values for four immunologic biomarkers widely used, 36 tegu lizards were evaluated tacking into account different age classes and both sexes. Total leukocyte counts were not different between age classes. Of the leucocytes count, eosinophils levels were higher in neonates compared with juvenile and adults; nevertheless, the heterophils group was the most prevalent leukocyte in the peripheral blood in all age classes. Lymphocytes, monocytes, heterophils, azurophils and basophils levels did not differ with age. Natural antibodies titres were higher in the adults compared with neonates and juveniles lizards. Lastly, complement system activity was low in neonates compared with juveniles and adults. Statistical analysis within each age group showed that gender was not a factor in the outcomes. Based on the results, we concluded that S. merianae demonstrated age (but not gender) related differences in the immune parameters analyzed. Having established baseline values for these four widely-used immunologic biomarkers, ongoing studies will seek to optimize the use of the S. merianae model in future research.

Mating System of Caiman yacare (Reptilia: Alligatoridae) Described from Microsatellite Genotypes.

The yacare caiman (Caiman yacare) is a reptile from South America and 1 of the 2 crocodilian species present in Argentina. The degradation of their natural habitat and strong hunting pressure led to a sharp numerical decline of wild populations. Nowadays, C. yacare is included in Appendix II of CITES, and ranching practices in some areas in Argentina are helping hatching success. In this context, it is important to better understand the population structure and mating system of the species. To do this, we amplified 10 microsatellite markers (SSRs) in 148 individuals of 13 C. yacare nests. All of the markers were polymorphic with 2­12 alleles per locus, with allelic sizes ranging between 154 and 400 bp and medium levels of polymorphism (Ho = 0.152­0.551 and He = 0.221­0.621). We were able to determine the maternal genotype in 9 out of 13 nests. In 6 of them we found more than 1 paternal genotype, with a maximum of 3 fathers for a single nest. This study is the first to provide evidence of multiple paternity behavior. These findings will be useful to improve management and conservation strategies for the species.

Latin American protected areas: Protected from chemical pollution?

Protected areas (PAs) are critically important means to preserve species and maintain natural ecosystems. However, the potential impacts of chemical pollution on PAs are seldom mentioned in the scientific literature. Research on the extent of the occurrence of chemical pollution inside PAs and in-depth assessments of how chemical contaminants may adversely affect the maintenance of species abundance, species survival, and ecosystem functions are scarce to nonexistent. We investigated 1) the occurrence of chemical contaminants inside 119 PAs in Latin America from publically available databases, and 2) reviewed case studies of chemical contaminants and pollution in 4 Latin American PAs. Cases of chemical pollution and contamination inside Latin American PAs mostly originated from sources such as mining, oil, and gas extraction. To date, the focus of the research on chemical pollution research inside Latin American PAs has been primarily on the detection of contamination, typically limited to trace metals. Where management actions have occurred, they have been reactive rather than proactive. Protected areas established in wetlands are the most affected by chemical pollution. Based on the information from the pollution and/or contamination occurrence and the case studies analyzed, Latin American PAs are not well safeguarded from chemical pollution, resulting in both challenges and opportunities to conserve biodiversity and ecosystems. Integr Environ Assess Manag 2017;13:360-370. © 2016 SETAC.

Total and differential white blood cell counts in Caiman latirostris after in ovo and in vivo exposure to insecticides.

Agricultural activities associated mainly with soybean crops affect the natural environment and wildlife by habitat destruction and the extensive use of agrochemicals. The aim of this study was to evaluate immunotoxic effects of the insecticides cypermethrin (CYP) and endosulfan (END) in Caiman latirostris analyzing total blood cell count (TWBC) and differential white blood cell count (DWBC) after in ovo and in vivo exposure. Eggs (in ovo) and hatchlings (in vivo) from nests harvested in natural habitats were artificially incubated and reared under controlled conditions in the Proyecto Yacaré (Gob.Santa Fe/MUPCN) facilities. Exposure of embryos was performed by topication on the eggshell during the first stage of development. The treatments were distilled water (negative control; NC), ethanol (vehicle control; VC), four groups treated with different concentrations of CYP and four groups with END. In vivo exposure was performed by immersion; treatments were NC, VC, two groups exposed to CYP and two to END. After embryonic exposure to the insecticides, no differences were found in TWBC or DWBC among the neonates exposed to pesticides versus controls. In the in vivo scenario, similar results were obtained for TWBC, but DWBC data showed differences between NC hatchlings and CYP-1 hosts for heterophil, lymphocyte and monocyte levels, and between NC and END-1 hosts for lymphocyte and monocyte levels. Research on the effects of pesticide exposure on this species is of special interest not only to assess the impact on caiman populations, but also to further characterize the species as a potential sentinel of ecosystem health.

Genotoxicity induced by Roundup® (Glyphosate) in tegu lizard (Salvator merianae) embryos.

Environmental contaminants produce multiple adverse consequences at individual, population and ecosystem levels. High volumes of agrochemicals applied to great variety of crops, together with agricultural expansion, generate great concerns due to the impact for the environment and large risk implicated for wildlife. The lack of data on these threats is striking. The tegu lizard (Salvator merianae) is one of the species that live in environments under contaminant effects. Several characteristics allow proposing this species as a potential sentinel organism for the monitoring of pesticides in their habitat. The present study is the first report about genotoxicity in tegu lizard neonates after embryonic exposure to Roundup® (glyphosate 66.2%). The micronucleus test (MN), nuclear abnormalities (NAs) assay and comet assay (CA) were used as biomarkers of genotoxic effects induced in erythrocytes by topical exposure of the eggs to the glyphosate commercial formulation Roundup® (RU), in laboratory controlled conditions. A total of 96 eggs were distributed in six groups exposed to RU (50, 100, 200, 400, 800, 1600µg/egg), one positive control (PC; 200µg cyclophosphamide/egg) and one negative control (NC; distilled water). No teratogenic effects were observed in any of the exposed or control neonates. A significant increase in DNA damage was observed in all concentrations higher than 100µg/egg with respect to NC (p<0.05). However, no statistical differences were found in the frequencies of MN and NAs in any group exposed to RU compared to the NC. No statistically significant differences were found in the size of the lizards at birth or after six months post-exposure (p>0.05). Our results provide new information about the undesirable effects of the glyphosate-based herbicide formulations RU on this lizard species that inhabits areas permanently exposed to several pesticide formulations. We consider of utmost necessity a strict regulation of the agrochemical application conditions in those environments near to places where wild populations of terrestrial and aquatic species live, in order to minimize the adverse effects on ecosystems.

Phenotypic variability and heritability of the cephalic region of Caiman latirostris.

The study of the cephalic shape of crocodilian is relevant in the fields of ecology, systematics, evolution, and conservation. Therefore, the integration of geometric analysis within quantitative genetics allows the evaluation of the inheritable shape components. In this study, the dorsal cephalic region of 210 Caiman latirostris hatchlings was analyzed from seven populations in Santa Fe, Argentina, to detect intra-, and inter-population phenotypic variability, and to determine the heritability of biological shape and size, using newly available geometric morphometric tools. The principal component analysis showed two configurations of cephalic shape that could be related to sexual dimorphism. In the canonical variate analysis, Procrustes distances between groups indicated that there are differences in shape among populations. Furthermore, the method of partial least squares indicated a covariation between cephalic shape and environmental variables. Regarding to CS of the skull we found significant differences among populations, moreover the partial least squares was also significant. Estimates of the heritability of shape and size were high, indicating that the components of these features are susceptible to the selection.

Immunotoxicity of commercial-mixed glyphosate in broad snouted caiman (Caiman latirostris).

The expansion and intensification of agriculture during the past 50 years is unprecedented, and thus environmental problems have been triggered at different scales. These transformations have caused the loss of habitat and biodiversity, and disruption of the structure and functioning of ecosystems. As a result of the expansion of the agricultural frontier in the recent past, many areas of the natural geographic distribution of the local wildlife, among them crocodilians and particularly the broad snouted caiman (Caiman latirostris), are being exposed to contaminants. The present study was designed to evaluate the effect of commercially-mixed glyphosate (RU) on some parameters of the immune system of C. latirostris. Two groups of caimans were exposed for two months to different concentrations of RU recommended for its application in the field, while one group was maintained as an unexposed control. The RU concentration was progressively decreased through the exposure period to simulate glyphosate degradation in water. After exposure, total and differential white blood cell (WBC), and complement system activity (CS) were determined. In addition, the animals were injected with a solution of lipopolysaccharide (LPS) from Escherichia coli to trigger an immune response and evaluate the parameters associated with it. The results showed that an effect of the herbicide on CS was observed, as animals exposed to RU showed a lower CS activity than animals from the negative control (NC) but not in total WBC. In the case of leukocyte population counts, differences were only found for heterophils and lymphocytes.

Spontaneous genetic damage in the tegu lizard (Tupinambis merianae): the effect of age.

Several studies indicate that certain factors such as age, sex or nutritional status among others, may affect the level of DNA damage, both induced and spontaneous, so it is very important to consider them for a more accurate interpretation of the findings. The aim of this study was to analyze the influence of age, sex, and nest of origin on spontaneous genetic damage of Tupinambis merianae determined by the comet assay (CA) and the micronucleus (MN) test, in order to improve reference data for future in vivo studies of xenobiotics exposure in this species. Sixty-five tegu lizards of three different ages: newborns (NB), juveniles (JUV) and adults (AD), both sexes and from different nests of origin were used. Blood samples were collected from the caudal vein of all animals and the MN test and CA were applied on peripheral blood erythrocytes to determine basal frequency of MN (BFMN) and basal damage index (BDI). The comparison between age groups showed statistically significant differences in the BFMN and BDI (p<0.05). NB animals showed significantly higher BDI values in relation to JUV and AD (p<0.016), but no statistically differences were found between the latter two. NB showed lower BFMN respect to other age groups, being statistically significant only when compared to AD (p<0.016). BFMN or BDI showed no statistically significant differences between sexes or nests of origin (p>0.05). A weak negative relationship was found only between BFMN and weight of NB tegu lizard (p=0.014; R(2)=0.245). Basal values of genetic damage obtained with both biomarkers in the tegu lizard evidenced that age is an intrinsic factor that should be taken into account to avoid misunderstanding of the results in future biomonitoring studies.

Detection and characterization of phospholipase A(2) (PLA(2)) in Caiman latirostris and Caiman yacare plasma.

Reptiles have proven to have a versatile and efficient nonspecific immune system adapted to the environments in which they commonly live. Phospholipase A(2) (PLA(2)) is important hydrolytic enzyme involved in the regulation of specific types of messengers, with significant roles in the innate immune response. A number of agents that exert effects on cellular receptors emit a series of signals leading to the increased activity of PLA(2). Phospholipase A(2) has been identified and characterized in temperature, plasma concentration, and kinetic dependence in two species of caiman. The results of these studies suggest that the high PLA(2) activities observed in caiman plasma may be an important component of a well-developed innate immunity. Based on the knowledge of their properties, this powerful immunologic component should be evaluated as a possible application in the veterinary or even human therapeutic industry. Additionally, this is another reason to consider these animals excellent models for the study of immune phylogenetic mechanisms.

Effects of in vivo exposure to Roundup® on immune system of Caiman latirostris.

The present study aimed to evaluate the effect of Roundup(®) (RU, glyphosate-based formulation) on some parameters of the immune system and growth of Caiman latirostris. Seventy-two caimans (20-day-old) from Proyecto Yacaré (Gob. Santa Fe/MUPCN) were used. Two groups were exposed for 2 months to different concentrations of RU (11 or 21 mg/L; taking into account the concentration recommended for its application in the field), while one group was maintained as control. The RU concentration was progressively decreased through the exposure period to simulate glyphosate degradation in water. Animals were measured and weighed at the beginning and end of the experiment, and blood samples taken after exposure to determine total and differential white blood cell (WBC) counts as well as total protein concentration (TPC), and for performing protein electrophoresis. The results showed that, compared against control hosts, there was a decrease in WBC counts, a higher percentage of heterophils, a higher TPC (with a low percentage of F2 protein fraction), and a negative effect on growth in the young caimans exposed to RU. These results demonstrate that in vivo exposure to RU induced alterations in the selected immune parameters, plasma proteins, and growth of caimans, thereby providing relevant information about the effects of this type of pesticide in this important species in the Argentinian wetlands.