Ecological factors shaping ectoparasite communities on heteromyid rodents at Médanos de Samalayuca.

Rodent ectoparasites are vectors for important pathogens of wildlife, domestic animals, and even zoonosis. Nevertheless, distribution patterns of ectoparasites are not fully understood; habitat, season, and host species are important predictors of distribution and prevalence. Heteromyid rodents are considered important reservoirs of diseases, given the presence of different ectoparasites and pathogens in them, and they offer the opportunity to learn about the ecology of parasites. The aim of the present work was to survey ectoparasites associated with heteromyid rodents near a National Protected Area in Chihuahua Mexico, south of the USA-Mexico border, and asses the effects of ecological factors (season, vegetation type, host species, and host body condition) on parasite infestation. We sampled five different locations from January 2018 to July 2022; 845 heteromyid rodents were examined and 49 fleas and 33 ticks were collected. Ectoparasites belonged to the Siphonaptera and Ixodida orders, including three families Ixodidae (Riphicephalus sanguineus), Pulicidae (Pulex irritans), and Ctenophthalmidae (Meringins altipecten, M. dipodomys). Five species of host rodents were captured, Dipodomys merriami, D. ordii, Chaetodipus eremicus, C. hispidus, and C. intermedius, but the last two species did not present any ectoparasites. Dipodomys merriami presented the highest flea and tick prevalence followed by D. ordii. We found parasitic partnerships between heteromyids according to ecological factors. The infestation in C. eremicus was related to body condition, vegetation type, and sex; in D. merriami, it was related to vegetation type and season, while D. ordii did not present a clear pattern of infestation. Our results suggest that the infestation patterns of heteromyid rodents in desert habitats are species dependent.

Population genetic structure of a Chihuahuan Desert endemic mammal, the desert pocket gopher, Geomys arenarius.

The biogeographic history of the Chihuahuan Desert is complex, driven by numerous physiographic events and climatic changes. This dynamic history would have influenced the flora and fauna of the region including the desert pocket gopher, Geomys arenarius, a subterranean rodent endemic to the northern Chihuahuan Desert. G. arenarius is restricted to sandy soils and are considered to have a disjunct distribution. Two subspecies are recognized: G. a. arenarius and G. a. brevirostris. We used multilocus nuclear (amplified fragment length polymorphisms) and mitochondrial DNA (ND2) sequence data to uncover patterns of genetic diversity within and among populations of G. arenarius. We evaluated correspondence of genetic patterns to traditionally accepted subspecies boundaries, mapped the distribution of potentially suitable soils to identify barriers or corridors to dispersal and to guide future survey efforts, provided evidence that could be used to recognize distinct population segments, and quantified genetic diversity within populations. Both datasets were largely concordant and demonstrated hierarchical patterns of genetic divergence. The greatest divergence was consistent with the two recognized subspecies. Mapping of potentially habitable soils revealed likely barriers to dispersal contributing to the allopatric pattern of geographic distribution and areas, which may be occupied by G. arenarius but not yet documented. Because G. arenarius is restricted to soils with high sand content, and these habitable soils are disjunct within the region occupied by this species, historical factors that impacted soil deposition and deflation likely contributed to the observed patterns of genetic divergence. Genetic diversity was higher within populations of the southern subspecies (G. a. arenarius) compared to G. a. brevirostris. This may be due to a greater availability of continuous suitable soils within the range of G. a. arenarius or higher density due to greater food availability (currently or historically)-both of which could allow for a higher effective population size.

Cytoprotective effects of creosote bush (Larrea tridentata) and Southern live oak (Quercus virginiana) extracts against toxicity induced by venom of the black-tailed rattlesnake (Crotalus ornatus).

The venom of Crotalus ornatus (vCo) poses a threat to human health, as it contains a mixture of toxins that can cause cytotoxic, necrotic, and hemolytic effects. The present study assessed methanolic and acetone extracts from leaves and flowers of Larrea tridentata, as well as the bark of Quercus virginiana as potential suppressors of the toxic effects of vCo in vitro. The content of total phenols, flavonoids, and tannins of the plant extracts were quantified for the suppression of vCo cytotoxicity in two cell culture models, human lymphocytes and porcine aortic endothelial (PAE) cells. Extracts from Q. virginiana displayed a greater concentration of total phenols, flavonoids, and tannins. Co-incubation of lymphocytes and PAE cells with fixed concentrations of vCo and plant extracts resulted in decreased vCo-induced cytotoxicity. A 24-hour co-incubation of lymphocytes with vCo (2.36 ± 0.17 µg/mL) and 0.5 µg/mL of methanolic leaf extract from L. tridentata (LLM) significantly suppressed the venom-induced cytotoxicity by 37.33 ± 8.33%. Similarly, the LLM extract (4 µg/mL) caused a significant decrease in vCo cytotoxicity after 24 hours in PAE cells. In contrast, while the acetone extract of Q. virginiana bark (QA) suppressed cytotoxicity by 29.20 ± 3.51% (p < 0.001) in lymphocytes, it failed to protect PAE cells against vCo after 24 hours. In PAE cells, a shorter 4-hour co-incubation showed significant suppression of cytotoxicity with both extracts. Our results collectively suggest that LLM and QA possess cytoprotective properties against the in vitro toxic effects of vCo, and thus establish extracts from these plants as potential therapeutic interventions against Crotalus envenomation.

New species of flea of the genus Dactylopsylla Jordan (Insecta: Siphonaptera) from the Flora and Fauna Protection Area Mdanos de Samalayuca, Chihuahua, Mexico.

A new species of Dactylopsylla Jordan, parasites of Geomyidae rodents from Chihuahua, is described and illustrated as D. samalayucae n. sp. This species is compared with their morphologically closest relatives. Males are characterized by the shape of the upper lobe of the inmovable and movable process, the shape and chaetotaxy of the distal arm of sternum IX and the shape of the crochet; and females by the contour of the distal margin of sternum VII. The geographical distribution of Dactylopsylla is extended to the Chihuahuan desert in Mexico as D. samalayucae n. sp. is reported from south of the Natural Protected Area Mdanos de Samalayuca (MS). A recent key to Dactylopsylla species is updated with inclusion of the new species.

Toxicological comparison of Crotalus ruber lucasensis venom from different ecoregions of the Baja California Peninsula.

The Baja California Peninsula possesses a mosaic of ecoregions that offers a wide variety of environments for the species that here inhabit. Here we report biological variations in. Crotalus ruber lucasensis venom from arid, semiarid and tropical eco-regions. Lethal (1.4-6.8 mg/kg), edematogenic (0.3-0.5 µg) and defibrinogenating (from non-detectable to 20 µg) activities were found to have significant differences among eco-regions.

Functional Mining of the Crotalus Spp. Venom Protease Repertoire Reveals Potential for Chronic Wound Therapeutics.

Chronic wounds are a major health problem that cause millions of dollars in expenses every year. Among all the treatments used, active wound treatments such as enzymatic treatments represent a cheaper and specific option with a fast growth category in the market. In particular, bacterial and plant proteases have been employed due to their homology to human proteases, which drive the normal wound healing process. However, the use of these proteases has demonstrated results with low reproducibility. Therefore, alternative sources of proteases such as snake venom have been proposed. Here, we performed a functional mining of proteases from rattlesnakes (Crotalus ornatus, C. molossus nigrescens, C. scutulatus, and C. atrox) due to their high protease predominance and similarity to native proteases. To characterize Crotalus spp. Proteases, we performed different protease assays to measure and confirm the presence of metalloproteases and serine proteases, such as the universal protease assay and zymography, using several substrates such as gelatin, casein, hemoglobin, L-TAME, fibrinogen, and fibrin. We found that all our venom extracts degraded casein, gelatin, L-TAME, fibrinogen, and fibrin, but not hemoglobin. Crotalus ornatus and C. m. nigrescens extracts were the most proteolytic venoms among the samples. Particularly, C. ornatus predominantly possessed low molecular weight proteases (P-I metalloproteases). Our results demonstrated the presence of metalloproteases capable of degrading gelatin (a collagen derivative) and fibrin clots, whereas serine proteases were capable of degrading fibrinogen-generating fibrin clots, mimicking thrombin activity. Moreover, we demonstrated that Crotalus spp. are a valuable source of proteases that can aid chronic wound-healing treatments.

In vitro hemotoxic, α-neurotoxic and vasculotoxic effects of the Mexican black-tailed rattlesnake (Crotalus molossus nigrescens) venom.

The Mexican black-tailed rattlesnake Crotalus molossus nigrescens is distributed in the Mexican plateau. Its venom is known to cause hemolysis and presents fibrinogen coagulase, collagenase and fibrinolytic activities. These activities may be associated with hemostatic alterations, such as platelet aggregation, hemolysis and fibrinolysis, often described in ophidic accidents. However, the mechanisms of action of the C. m. nigrescens venom remain unclear. In this study we investigated the in vitro hemotoxic, neurotoxic, and vasculotoxic effects of the venom. We found that this venom produces two types of hemolytic responses, Oxyhemoglobin release and Methemoglobin formation. As a result of the cytotoxicity to endothelial cells produces morphological biphasic toxicity. The first step in this process is characterized by morphological changes, as well as the loss of cellular adhesion and reduction in thickness. The second phase is characterized by massive cellular aggregation and death. It also induced laminin, type IV collagen, perlecan and nidogen degradation. However, the venom did not modulate the muscular fetal and neuronal nicotinic acetylcholine receptors activity. Thus, we concluded that the C. m. nigrescens venom produced hemolysis and hemorrhages via degradation of the basement membrane components and endothelial cell cytotoxicity, but not by neurotoxicity at the receptor level in nicotinic acetylcholine receptors.

Rattlesnake Crotalus molossus nigrescens venom induces oxidative stress on human erythrocytes.

BACKGROUND: Globally, snake envenomation is a well-known cause of death and morbidity. In many cases of snakebite, myonecrosis, dermonecrosis, hemorrhage and neurotoxicity are present. Some of these symptoms may be provoked by the envenomation itself, but others are secondary effects of the produced oxidative stress that enhances the damage produced by the venom toxins. The only oxidative stress effect known in blood is the change in oxidation number of Fe (from ferrous to ferric) in hemoglobin, generating methemoglobin but not in other macromolecules. Currently, the effects of the overproduction of methemoglobin derived from snake venom are not extensively recorded. Therefore, the present study aims to describe the oxidative stress induced by Crotalus molossus nigrescens venom using erythrocytes. METHODS: Human erythrocytes were washed and incubated with different Crotalus molossus nigrescens venom concentrations (0-640 µg/mL). After 24 h, the hemolytic activity was measured followed by attenuated total reflectance-Fourier transform infrared spectroscopy, non-denaturing PAGE, conjugated diene and thiobarbituric acid reactive substances determination. RESULTS: Low concentrations of venom (<10 µg/mL) generates oxyhemoglobin release by hemolysis, whereas higher concentrations produced a hemoglobin shift of valence, producing methemoglobin (>40 µg/mL). This substance is not degraded by proteases present in the venom. By infrared spectroscopy, starting in 80 µg/mL, we observed changes in bands that are associated with protein damage (1660 and 1540 cm-1) and lipid peroxidation (2960, 2920 and 1740 cm-1). Lipid peroxidation was confirmed by conjugated diene and thiobarbituric acid reactive substance determination, in which differences were observed between the control and erythrocytes treated with venom. CONCLUSIONS: Crotalus molossus nigrescens venom provokes hemolysis and oxidative stress, which induces methemoglobin formation, loss of protein structure and lipid peroxidation.

Rattlesnake Crotalus molossus nigrescens venom induces oxidative stress on human erythrocytes

Abstract Background Globally, snake envenomation is a well-known cause of death and morbidity. In many cases of snakebite, myonecrosis, dermonecrosis, hemorrhage and neurotoxicity are present. Some of these symptoms may be provoked by the envenomation itself, but others are secondary effects of the produced oxidative stress that enhances the damage produced by the venom toxins. The only oxidative stress effect known in blood is the change in oxidation number of Fe (from ferrous to ferric) in hemoglobin, generating methemoglobin but not in other macromolecules. Currently, the effects of the overproduction of methemoglobin derived from snake venom are not extensively recorded. Therefore, the present study aims to describe the oxidative stress induced by Crotalus molossus nigrescens venom using erythrocytes. Methods Human erythrocytes were washed and incubated with different Crotalus molossus nigrescens venom concentrations (0640 g/mL). After 24 h, the hemolytic activity was measured followed by attenuated total reflectance-Fourier transform infrared spectroscopy, non-denaturing PAGE, conjugated diene and thiobarbituric acid reactive substances determination. Results Low concentrations of venom ( 10 g/mL) generates oxyhemoglobin release by hemolysis, whereas higher concentrations produced a hemoglobin shift of valence, producing methemoglobin (>40 g/mL). This substance is not degraded by proteases present in the venom. By infrared spectroscopy, starting in 80 g/mL, we observed changes in bands that are associated with protein damage (1660 and 1540 cm1) and lipid peroxidation (2960, 2920 and 1740 cm1). Lipid peroxidation was confirmed by conjugated diene and thiobarbituric acid reactive substance determination, in which differences were observed between the control and erythrocytes treated with venom. Conclusions Crotalus molossus nigrescens venom provokes hemolysis and oxidative stress, which induces methemoglobin formation, loss of protein structure and lipid peroxidation.

Rattlesnake Crotalus molossus nigrescens venom induces oxidative stress on human erythrocytes

Background Globally, snake envenomation is a well-known cause of death and morbidity. In many cases of snakebite, myonecrosis, dermonecrosis, hemorrhage and neurotoxicity are present. Some of these symptoms may be provoked by the envenomation itself, but others are secondary effects of the produced oxidative stress that enhances the damage produced by the venom toxins. The only oxidative stress effect known in blood is the change in oxidation number of Fe (from ferrous to ferric) in hemoglobin, generating methemoglobin but not in other macromolecules. Currently, the effects of the overproduction of methemoglobin derived from snake venom are not extensively recorded. Therefore, the present study aims to describe the oxidative stress induced by Crotalus molossus nigrescens venom using erythrocytes. Methods Human erythrocytes were washed and incubated with different Crotalus molossus nigrescens venom concentrations (0-640 μg/mL). After 24 h, the hemolytic activity was measured followed by attenuated total reflectance-Fourier transform infrared spectroscopy, non-denaturing PAGE, conjugated diene and thiobarbituric acid reactive substances determination. Results Low concentrations of venom (<10 μg/mL) generates oxyhemoglobin release by hemolysis, whereas higher concentrations produced a hemoglobin shift of valence, producing methemoglobin (>40 μg/mL). This substance is not degraded by proteases present in the venom. By infrared spectroscopy, starting in 80 μg/mL, we observed changes in bands that are associated with protein damage (1660 and 1540 cm−1) and lipid peroxidation (2960, 2920 and 1740 cm−1). Lipid peroxidation was confirmed by conjugated diene and thiobarbituric acid reactive substance determination, in which differences were observed between the control and erythrocytes treated with venom. Conclusions Crotalus molossus nigrescens venom provokes hemolysis and oxidative stress, which induces methemoglobin formation, loss of protein structure and lipid peroxidation.(AU)

Capillary damage in the area postrema by venom of the northern black-tailed rattlesnake (Crotalus molossus molossus).

The Northern black-tailed rattlesnake (Crotalus molossus molossus) venom is mainly hemotoxic, hemorrhagic, and neurotoxic. Its effects in the central nervous system are unknown and only poorly described for all Viperidae species in general. This is why we are interested in describe the damage induced by C. m. molossus venom in rat brain, particularly in the area postrema capillaries. Four C. m. molossus venom doses were tested (0.02, 0.05, 0.10 and 0.20mg/kg) injected intramuscularly at the lower limb, incubated by 24 hours and the brains were harvested. Area postrema coronal sections were stained with Haematoxylin and Eosin, and examined to observe the venom effect in quantity of capillaries and porphology. Starting from the 0.10mg/kg treatment we observed lysed extravasated erythrocytes and also capillary breakdown, as a consequence of hemorrhages appearance. The number of capillaries decreased significantly in response to the venom dose increment. Hemorrhages could be caused by the metalloproteinase activity on the basal membrane and the apoptosis generated by L-amino acid oxidases. Hemolysis could be caused by phospholipase A2 hemotoxic effect. We conclude that C. m. molossus crude venom produces hemolysis, capillary breakdown, hemorrhages, and the reduction in number of capillaries in the area postrema.