Distribution and ecological risk of metals in an urban natural protected area in the Riviera Maya, Mexico.

Urbanization can negatively impact natural protected areas near or surrounded by cities, and such impacts include untreated wastewater discharge, leachates from dumpsters, e-waste, and road dust. In this research, we show that not only large cities with industry are prone to be polluted, but also young touristic cities with high population increase rate can suffer from urban contamination. We evaluated metal pollution in a natural protected area within a 50-year-old city without conventional industry that was likely contaminated by the urban sprawl around the protected area. We tested water, zooplankton, sediment and plant samples for metallic elements to evaluate their bioaccumulation in zooplankton, enrichment factors and geoaccumulation index values in sediments, and translocation factors in plants. Finally, we evaluated the ecological risk due to metal contamination. Metals at levels above our detection limit (20 µg/L) were not found in the water and zooplankton samples. The sediments and plants in the storm drain section of the protected area had a greater concentration of metals and wastewater indicators (coliforms) than those in the rest of the lagoon. Moreover, signs of Al, Cu, Ni, Zn, Cr, Pb, and Ti contamination were found in the plant tissues. We estimated that the ecological risk of this natural protected area surrounded by the city of Cancun (Mexico) ranged from mild to strong, with Zn being the metal of most concern. The results highlight that young touristic cities around the world will endure contamination from urban sources; signs or early warnings of contamination must be identified to prevent and resolve such issues.

Wound healing activity of the hydroalcoholic extract and the main metabolites of Amphipterygium adstringens (cuachalalate) in a rat excision model.

ETHNOPHARMACOLOGICAL RELEVANCE: The bark of Amphipterygium adstringens Schiede ex Schltdl (Anacardiaceae), commonly known as 'cuachalalate' has been used in Mexican traditional medicine for the treatment of skin and oral lesions, gastric ulcers, and other conditions. The use as wound healing of the bark of this plant has been known since before the Spanish conquest of Mexico. Its uses are mentioned in the first writings of the Spanish in the 16th century. It is important to highlight that its use for wound healing treatment has no scientific previous reports. AIM OF THE STUDY: The objectives of this study were to determine the wound healing effect of the hydroalcoholic extract of the stem bark of Amphipterygium adstringens and its main metabolites, using a model of excision in the back of Wistar rats. To evaluate its antimicrobial effect against common bacteria that living on the skin of wounds and to evaluate its effect on angiogenesis. MATERIALS AND METHODS: The hydroalcoholic extract of cuachalalate (HAE, 10 mg/wound/day), the 3α-hydroxymasticadienoic acid (3 MA, 300 µg/wound/day), the masticadienoic acid (MA, 300 µg/wound/day), and a mixture of anacardic acids (ANA, 300 µg per wound) were tested in a murine excision model topically for 15 days, to evaluate their wound-healing effect. The results were reported in a wound closure percentage (n = 30 animals per treatment curve), using pirfenidone (PIR, 8% in vehicle) as a reference drug. In addition, histologic analysis was performed to evaluate the structure and quality of the scar. The effect on angiogenesis was assessed using the chick embryo chorioallantoic membrane (CAM) model (n = 6 eggs per treatment). The concentration evaluated for each treatment was 300 µg, using as proangiogenic reference drug the histamine (HIS, 5.6 µg) and as antiangiogenic drugs pirfenidone (9 µg) and acetylsalicylic acid (ASA, 9 µg). The antimicrobial test was performed against S. mutans, S. aureus, P. aeruginosa y E. coli using a minimum inhibitory concentration (MIC) assay. RESULTS: The 3α-hydroxymasticadienoic (3 MA) acid and the anacardic acids (ANA) improve the wound closure by approximates 30% (similar to pirfenidone) in comparison with the control-treated with the vehicle in the proliferative phase. On the other hand, the hydroalcoholic extract of cuachalalate (HAE) did not show an effect on the wound healing process. The histologic analysis demonstrated that the three main metabolites showed an improvement in the scar structure. According to the CAM results, it is probable that the main action mechanism of the 3α-hydroxymasticadienoic acid and the anacardic acids is related to their proangiogenic effect. In addition, ANA showed a modest antimicrobial effect. CONCLUSIONS: The 3α-hydroxymasticadienoic acid and anacardic acids showed a better tissue structure and reduced the time closure of the wound. In addition, the anacardic acids showed antimicrobial effects and both metabolites promote angiogenesis, suggesting that these effects may be related to their action mechanism. These metabolites of cuachalalate could be a good alternative for wound healing treatment.

Cadmio y alteraciones morfológicas en el rotífero Philodina cf. roseola (Bdelloidea: Philodinidae) y el gusano Aeolosoma hemprichi (Annelida: Aeolosomatidae)

El cadmio es un metal tóxico que produce deformaciones en el zooplancton y es considerado un peligro para el ambiente y la vida acuática. Por ello, utilizamos dos especies de zooplancton nativas de Quintana Roo, México para obtener datos sobre la toxicidad de cadmio, incluyendo la concentración umbral para alteraciones morfológicas observables y el porcentaje de organismos con alteraciones morfológicas en las concentraciones de exposición. Utilizamos el rotífero Philodina cf roseola y el oligoqueto Aeolosoma hemprichi, ya que ambos se alimentan del alga Nannochloropsis oculata. Seguidamente, ambas especies de animales fueron expuestas a un rango de concentración de cadmio de 0.05 (0.047 mg/l, concentración real) a 10.0 mg/l (9.398 mg/l, concentración real) durante 24 h. El CL50 para cadmio en P. cf roseola fue de 0.7 mg/l (0.65 mg/l, concentración real), mientras que en A. hemprichi fue de 3.38 mg/l (3.17 mg/l, concentración real). El cadmio a concentraciones de 0.5 mg/l (0.47 mg/l, concentración real) y exposición menor a 24 h, por un lado, indujo alteraciones morfológicas en la lorica de los rotíferos, deformaciones del pie y constricciones en la parte media del cuerpo. En oligoquetos, por otro lado, observamos daños en la epidermis. La concentración umbral donde las deformaciones aparecieron en rotíferos fue 0.3 mg/l (0.28 mg/l, concentración real) a las 24 h, mientras que en oligoquetos fue a 0.5 mg/l (0.47 mg/l concentración real) en 30 min. Concluimos que claramente el cadmio promueve alteraciones morfológicas en organismos acuáticos, debido a la exposición directa que se añade al medio de cultivo.

Cadmium is a toxic metal for zooplankton that produces deformations. It is also considered an environmental hazard to aquatic life. Since it has a significant effect in some marine organisms, we used two native zooplankton species from Quintana Roo, Mexico to obtain data regarding cadmium toxicity including the threshold concentration for observable morphological alterations and the percentage of organisms with morphological alterations at the exposure concentrations. We used the rotifer Philodina cf roseola and the oligochaeta Aeolosoma hemprichi, since both feed from the algae Nannochloropsis oculata. Both animals were exposed to a cadmium concentration range from 0.05 mg/l (0.047 mg/l, real concentration) to 10.0 mg/l (9.39 mg/l, real concentration) for 24 h. The LC50 for cadmium in P. cf roseola was 0.7 mg/l (0.65 mg/l, real concentration), whereas in A. hemprichi was 3.38 mg/l (3.17 mg/l, real concentration). The exposure of cadmium at 0.5 mg/l (0.47 mg/l, real concentration) for less than 24 h induced morphological alterations in the lorica of rotifers, foot deformations, and constriction in the middle part of the body. In oligochaetes, damage to the epidermis was noted. The threshold concentration where deformations appeared in rotifer was 0.3 mg/l at 24 h, while in oligochaeta was 0.5 mg/l (0.47 mg/l, real concentration) at 30 min. We concluded that cadmium promotes alterations in aquatic organisms due to direct exposure added to the culture medium.

Monogonot rotifers species of the island Cozumel, Quintana Roo, México.

The current study adds 23 new records to the previously described monogonot rotifers of Cozumel, increasing the number of reported species to 40; these species are grouped into two orders, 11 families and 18 genera. Littoral and limnetic samples from 17 aquatic systems were examined, including wet dolines, coastal lagoons, temporary pools and artificial systems (reservoirs). Of the 36 species found, the following organisms represent new findings for Cozumel: Anuraeopsis fissa, Brachionus angularis, B. rubens, B. plicatilis sensu lato, Beuchampiella eudactylota, Euchlanis dilatata, Mytilina bisulcata, Colurella adriatica, Lepadella ovalis, L. rhomboides, Squatinella mutica, Lecane aculeata, L. climacois, L. crepida, L. cornuta, L. grandis, L. obtusa, L. pyriformis, Cephalodella forficula, Scaridium bostjani, Trichocerca pusilla, Polyarthra cf. dolichoptera, P. vulgaris, Dicranophorus epicharis and Testudinella patina. Additional information is reported for these species.

Adverse effects and bioconcentration of chromium in two freshwater rotifer species.

Bioaccumulation of trivalent (CrIII) and hexavalent chromium (CrVI), and its adverse effects were studied in two rotifer species: Brachionus calyciflorus (two different strains), and Lecane quadridentata. Median Lethal Concentration (LC50) at 24 h of both species showed that CrVI is highly toxic: LC50 ranges from 4.7 × 10(-5) to 4 × 10(-6) mg L(-1)), compared with CrIII: LC50 ranges from 0.64 to 1.279 mg L(-1). Using the LC50 as an exposure concentration, and using atomic absorption, the bioconcentration factor (BCF) was obtained and BCFs of rotifers exposed to CrIII are four orders of magnitude lower than BCFs of rotifers exposed to CrVI. The effect of Cr on the elemental composition of the two species of rotifers in their structures by X-ray microanalysis by energy dispersion showed that Cr is found in intoxicated rotifers, but not in control rotifers. The basal immunoreactivity to metallothioneins is greater in B. calyciflorus than L. quadridentata. The immunoreactivity to metallothioneins decreases in B. calyciflorus when is exposed to CrIII, in contrast in L. quadridentata the immunoreactivity to metallothioneins increase when is exposed to CrIII, and the immunoreactivity to CrVI in L. quadridentata decrease. A mechanism is proposed in which the harder lorica of L. quadridentata acts as a barrier and accumulator of CrVI, and allows for attenuating responses like metallothionein production in L. quadridentata. Instead, in B. calyciflorus the lack of a harder lorica allows for deeper penetration of CrVI, and no time to produce attenuating measures.

Morphological alterations in the freshwater rotifer Brachionus calyciflorus Pallas 1766 (Rotifera: Monogononta) caused by vinclozolin chronic exposure.

Vinclozolin (VZ) is a dicarboximide fungicide widely used on fruits, vegetables and wines, effective against fungi plagues. In this study we characterized the effects of VZ using a 4-day reproductive chronic assay with the freshwater rotifer Brachionus calyciflorus. The assay included observations of several features of asexual and sexual reproduction. Our results indicate that VZ: (a) increased asexual and sexual reproduction, (b) caused severe abnormality in females and (c) these abnormalities were inherited by sexual and asexual reproduction. At 1.2 mg/L three abnormal females were found out of 457 total females (0.66 %). This low percentage is consistent and reproducible according to further analysis, where we increased the number of replicates and total females exposed to 1.2 mg/L of VZ, and found 18 abnormal females out of 2868 total females (0.63 % abnormality). Interestingly, abnormal females found at 5.6 mg/L VZ exposure, were able to show mating behavior. Our results suggest that VZ behaves as a strong endocrine disruptor whose effects show the characteristic inverted-U-shape exposure concentration response curve regarding the intrinsic population increase and the percentage of abnormalities as endpoints.

Bioconcentration and localization of lead in the freshwater rotifer Brachionus calyciflorus Pallas 1677 (Rotifera: Monogononta).

We studied how lead is bioconcentrated and distributed in the rotifer Brachionus calyciflorus using metal histochemistry to locate lead granules, Leadmium Green® analysis to establish the route of uptake, atomic absorption to determined the bioconcentration factor (BCF), and detected the presence of microelements in the cuticle by X-ray microanalysis with scanning electron microscopy. Our results indicate: (a) the digestive system is the main route of lead uptake in the rotifer B. calyciflorus, (b) after 24-h lead is deposited in granules in the mastax and vitellarium, (c) our energy-dispersive X-ray microanalysis indicates decalcification taking place in the cuticle of the rotifer after a 24-h lead exposure, and (d) we determined a BCF = 115 for lead after a 24 h exposure. However, the route of mobilization and storage of intracellular lead are still not fully understood in B. calyciflorus.

Immunodetection of Luteinizing Hormone (LH), Follicle-Stimulating Hormone (FSH), Thyroid Stimulating Hormone (TSH) and Prolactin (PRL) in Brachionus calyciflorus (Rotifera: Monogononta)

The endocrine system controls and coordinates behavioral, biochemical, and physiological processes through signal mechanisms using neuropeptides or products of neurosecretory cells. Among invertebrates, this system is poorly studied in rotifers, in which estrogens and androgens significantly affect sexual reproduction. This is the first report of the presence of the Luteinizing Hormone (LH), Follicle-Stimulating Hormone (FSH), Thyroid Stimulating Hormone (TSH) and Prolactin (PRL) in rotifers. Analyses included the avidin-biotin-peroxidase complex method with primary antibodies LH (Anti-Rat LH serum for RIA), PRL (Anti-Rat PRL serum for RIA), FSH (Anti-Rat FSH serum for RIA) and TSH (Anti-Rat TSH serum for RIA). These hormones were found in females, males and parthenogenetic and sexual eggs of the freshwater Brachionus calyciflorus. The immunoreactivity of FSH, LH, TSH and PRL in females was observed in: ovaries, cerebrum, mastax, stomach, lorica, and the stomach gland. However, in males LH was observed only at the trochal disk and cerebrum. The hormones FSH, TSH and PRL, were observed in testicles, contractil vesicles, and cementary gland of males. Regarding amictic or parthenogenetic eggs, the hormones LH, FSH, TSH, and PRL were located mainly in the micromeres, and the staining in the macromeres was weak. On the other hand, in the mictic or sexual eggs the inner shell is stained for the hormones PRL and LH, opposite to the staining of FSH and TSH, located mainly in the embryo. In general, immuno-reactivity was observed in areas important for the reproductive, excretory, digestive and developmental processes. Rev. Biol. Trop. 57 (4): 1049-1058. Epub 2009 December 01.

Se logró detectar la presencia de las hormonas: Hormona Luteinizante (LH), Hormona Folículo Estimulante (FSH), Hormona Estimulante de la Tiroides (TSH) y Prolactina (PRL) en Brachionus calyciflorus siendo el primer reporte de la presencia de dichas hormonas en rotíferos. Estas hormonas fueron identificadas por un método inmunológico-histológico-químico usando el complejo avidina-biotina- peroxidasa con los siguientes anticuerpos primarios: LH (Anti-Rata LH suero para RIA), PRL (Anti-Rata PRL suero para RIA), FSH (Anti-Rata FSH suero para RIA) y TSH (Anti-Rata TSH en suero para RIA). Estas hormonas se encontraron en las hembras, machos, huevos partenogenéticos y huevos sexuales del rotífero dulceacuícola B. calyciflorus. La reactividad inmunológica de FSH, LH, PRL y TSH en las hembras se observó en ovarios, cerebro, mástax, estómago, lorica, y la glándula del estómago. Sin embargo, en machos, la LH se observó sólo en el disco trocal y cerebro mientras que las hormonas FSH, PRL y TSH, se observaron en testículos, vesícula contráctil, y la glándula cementaria. En cuanto a los huevos partenogenéticos o amícticos, las hormonas LH, FSH, TSH, y PRL, se encontraron principalmente en los micrómeros, y en los macrómeros la tinción es débil. Por otra parte, el huevo sexual o míctico muestra reactividad inmunológica en la cubierta interior del huevo para las hormonas LH y PRL, lo contrario para FSH y TSH, las cuales se observaron principalmente en el embrión. La reactividad inmunológica fue observada, en general, en áreas importantes para los procesos reproductivos, excretorios, digestivos y del desarrollo.

Immunodetection of luteinizing hormone (LH), follicle-stimulating hormone (FSH), thyroid stimulating hormone (TSH) and prolactin (PRL) in Brachionus calyciflorus (Rotifera: Monogononta).

The endocrine system controls and coordinates behavioral, biochemical, and physiological processes through signal mechanisms using neuropeptides or products of neurosecretory cells. Among invertebrates, this system is poorly studied in rotifers, in which estrogens and androgens significantly affect sexual reproduction. This is the first report of the presence of the Luteinizing Hormone (LH), Follicle-Stimulating Hormone (FSH), Thyroid Stimulating Hormone (TSH) and Prolactin (PRL) in rotifers. Analyses included the avidin-biotin-peroxidase complex method with primary antibodies LH (Anti-Rat LH serum for RIA), PRL (Anti-Rat PRL serum for RIA), FSH (Anti-Rat FSH serum for RIA) and TSH (Anti-Rat TSH serum for RIA). These hormones were found in females, males and parthenogenetic and sexual eggs of the freshwater Brachionus calyciflorus. The immunoreactivity of FSH, LH, TSH and PRL in females was observed in: ovaries, cerebrum, mastax, stomach, lorica, and the stomach gland. However, in males LH was observed only at the trochal disk and cerebrum. The hormones FSH, TSH and PRL, were observed in testicles, contractil vesicles, and cementary gland of males. Regarding amictic or parthenogenetic eggs, the hormones LH, FSH, TSH, and PRL were located mainly in the micromeres, and the staining in the macromeres was weak. On the other hand, in the mictic or sexual eggs the inner shell is stained for the hormones PRL and LH, opposite to the staining of FSH and TSH, located mainly in the embryo. In general, immuno-reactivity was observed in areas important for the reproductive, excretory, digestive and developmental processes.