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There is little information in the scientific literature concerning sheep pregnancy and lambing success with regard to the timeframe from when in vitro produced embryos are transported to the designated location for embryo transfer (ET). The aim of this study was to transfer in vitro produced embryos under two different conditions that could typically occur using the aforementioned assisted reproductive techniques (ARTs). Abattoir ovaries were used to procure oocytes for in vitro embryo production and subsequent transfer to synchronized ewes. The study consisted of two experiments: Experiment 1 (Exp1)—embryos taken from the laboratory to a nearby surgical room for immediate ET, and Experiment 2 (Exp2)—ET after 5 hours (h) of transport to a rural farm. Lambing in relation to detected pregnancies, births compared to pregnancies, and the proportion of twin offspring were all higher in Exp2. Notably, in both Exp1 and Exp2, there was not a significant difference (P > 0.05) between the number of embryos transferred, i.e., 3 versus 4, respectively, and the number of ewes that underwent parturition in each group. Also, in both experiments there was not a significant difference (P > 0.05) in the number of ewes that underwent parturition based on the number (i.e., ≥1) of corpora lutea present. The results of the present study demonstrate the importance of evaluating different conditions when applying ARTs, as there are many variables that can influence the outcome. Importantly, Exp2 results show that ovine ET in places located far away from the embryo production site can be useful and successful provided that embryo transport, ET, and recipient conditions are adequate.

The objective of this study was to estimate crossbreeding effects for individual survival time (STIM) in hours, and survival rate for the complete experimental period (SUEX) during challenges with White Spot Disease (WSD), as well as for individual survival rate (SURP), harvest weight at 130 days of age (HW130) and biomass production per seeded shrimp (BIOM) estimated as SURP × HW130, in commercial-like ponds with (+) or without (−) disease outbreaks in Pacific white shrimp (Litopenaeus vannamei) on the Northwest coast of Mexico. Data was obtained from 2014 to 2016 from a crossbreeding experiment involving a line of Ecuadorian shrimp with a history of WSD resistance (Resistance Line – RES) and a Mexican line with a high growth potential (Growth Line – GRO), obtained by selection for a higher growth rate in the absence of specific diseases. For challenge tests, family-marked animals were exposed per os to minced muscle tissue from WSD diseased animals and followed until mortality stabilized. Families and organisms assessed in the three years of the experimental challenges for RES, GRO and crossbreeds were 149 and 4,705, 196 and 6,186 and 144 and 4,807, respectively. Similar figures for field data were 149 and 11,490, 196 and 10,927 and 145 and 12,246, respectively. Both challenge and field data were analyzed by linear models to estimate crossbreeding effects and predicted averages for pure lines and crossbreeds. Significant direct line effects (P

Moribund shrimp affected by acute hepatopancreatic necrosis disease (AHPND) from farms in northwestern Mexico were sampled for bacteriological and histological analysis. Bacterial isolates were molecularly identified as Vibrio parahaemolyticus by the presence of the tlh gene. The tdh -negative, trh -negative, and tlh -positive V. parahaemolyticus strains were further characterized by repetitive extragenic palindromic element-PCR (rep-PCR), and primers AP1, AP2, AP3, and AP and an ems2 IQ2000 detection kit (GeneReach, Taiwan) were used in the diagnostic tests for AHPND. The V. parahaemolyticus strains were used in immersion challenges with shrimp, and farmed and challenged shrimp presented the same clinical and pathological symptoms: lethargy, empty gut, pale and aqueous hepatopancreas, and expanded chromatophores. Using histological analysis and bacterial density count, three stages of AHNPD (initial, acute, and terminal) were identified in the affected shrimp. The pathognomonic lesions indicating severe desquamation of tubular epithelial cells of the hepatopancreas were observed in both challenged and pond-infected shrimp. The results showed that different V. parahaemolyticus strains have different virulences; some of the less virulent strains do not induce 100% mortality, and mortality rates also rise more slowly than they do for the more virulent strains. The virulence of V. parahaemolyticus strains was dose dependent, where the threshold infective density was 10 4 CFU ml −1 ; below that density, no mortality was observed. The AP3 primer set had the best sensitivity and specificity. Field and experimental results showed that the V. parahaemolyticus strain that causes AHPND acts as a primary pathogen for shrimp in Mexico compared with the V. parahaemolyticus strains reported to date.

Genotoxic, teratogenic, and cytotoxic chemical agents are often products of anthropogenic activities, such as agriculture and industry, and many eventually settle in marine and coastal ecosystems. Therefore, there is a need to implement monitoring programs that provide early warnings of the risks and problems associated with contamination to contribute to the conservation of coastal ecosystems and human health. This paper describes the fundamental characteristics of a micronucleus assay that uses seabird embryo blood and highlights its ability to detect the presence of micronucleogenic genotoxic agents along the coasts of Mexico. This paper also demonstrates the potential of this assay to assess possible seasonal and spatial effects of these agents on wildlife, which has direct implications for human health.

La Iniciativa Salud Mesoamerica (ISM) es un ambicioso esfuerzo regional de cooperacion basada en resultados que busca vincular una parte de los aportes efectuados por los donantes con la obtencion de determinados resultados en materia de salud reproductiva, materna, del recien nacido y del nino – tanto a nivel micro/prestacion de servicios como a nivel macro/poblacion – en los municipios mas pobres de los paises participantes. La ISM recurre unicamente a equipos independientes (externos) para medir aquellos resultados que determinan la aprobacion del financiamiento. En contraste, otras iniciativas de financiamiento basado en resultados generalmente determinan el pago mediante resultados autoinformados que son posteriormente verificados. Los paises se han beneficiado con este sistema ya que la medicion independiente y el apoyo de la ISM han ayudado a aumentar la capacidad local de recolectar y analizar datos con el fin de monitorear el desempeno, identificar cuellos de botella y pedir cuentas por los resultados. La razon principal de confiar en mediciones independientes obedecio a la exigencia de los donantes de condicionar los pagos por desempeno a los paises participantes a la obtencion de resultados reales y al requisito planteado por el Banco Interamericano de Desarrollo de minimizar el riesgo fiduciario asociado con la administracion de los fondos de los donantes. Otra de las razones de preferir las mediciones independientes fue la necesidad de contar con datos comparables en todos los paises para asi monitorear el impacto general y contribuir al aprendizaje global.

Salud Mesoamerica Initiative (SMI) is a unique results-based aid initiative that rewards countries for achieving health targets in the poorest municipalities in Mesoamerica. A partnership between private and public donors and governments, it offers lessons for philanthropists, corporate social responsibility teams, and bilateral donors about how to alter development assistance from the usual model of paying for inputs to a new one in which countries are paid for achieving results for their poorest populations. What makes this model of RBA feasible is its reliance on the staff, relationships, systems, and implementation capability of the Inter-American Development Bank and the dedicated unit that supports countries. Countries commit to achieve targets that progress from system readiness metrics to outcomes. Verification is based on externally conducted household and facility surveys. Five out of the eight countries achieved first-phase targets and received a performance payment, and all countries improved on key health system metrics.

Fish consumption is the major source of mercury to humans. Fetuses and children are the most sensitive groups of populations to the effects of mercury. Consequently, fish consumption by pregnant women, children, and women of childbearing age is of concern because of the effects of mercury exposure on human health. To assess mercury exposure in a population in northwest Mexico, the allowed daily consumption of fish (which indicates the maximum daily amount of fish that can be consumed without causing adverse noncarcinogenic effects) was calculated for the general population (GP) and fishing-related population (FRP). The studied groups for both sectors of the population were children A (3-10 years old), children B (11-15 years old), women of childbearing age (16-40 years old), and the rest of the population (men ≥16 years old, and women ≥41 years old). Mercury content in canned and frozen tuna, smoked marlin, tilapia, Pacific sierra, dolphinfish, and bullseye puffer ranged from 0.01 to 0.23 μg/g wet weight; none of the values were above the limit set by Mexico. Regarding mercury concentrations and rates of fish consumption, the GP consumes 1.7-2.7 times the allowed daily consumption, and the FRP consumes 1.6-3.9 that limit. The risk analysis showed the children A and B groups from the GP and adults of FRP to be the highest percentage of the population at risk (approximately 35%). These results highlight the need for adequate strategies that consider mercury exposure as part of public health policies associated with fish consumption in Mexico.

The state of Sinaloa in Mexico is an industrialized agricultural region with a documented pesticide usage of 700 t year−1; which at least 17 of the pesticides are classified as moderately to highly toxic. Pollutants in the water column of rivers and drains are of great concern because the water flows into coastal lagoons and nearshore waters and thereby affects aquatic organisms. This study was done in four municipalities in the state of Sinaloa that produce food intensively. To investigate the link between pollution in the lagoons and their proximity to agricultural sites, water was sampled in three coastal lagoons and in the rivers and drains that flow into them. Seawater from the Gulf of California, 10 km from the coast, was also analyzed. Concentrations of nutrients, organochlorines, and organophosphorus pesticides were determined. Nutrient determination showed an unhealthy environment with N/P ratios of

For years our efforts have been focused on vaccination during childhood. Today we know that this is not enough to ensure health in the rest of the life. Childhood is as important as any other stage and, therefore, vaccination must be permanent and differentiated, according to our age, throughout life. Introducing a life course perspective in vaccination programs, with emphasis on adult vaccination, particularly in older adults, offers us the opportunity to review the performance of health programs, actions, and services in the field of immunization, as well as strengthening health promotion actions. In this context, the first Mexican Consensus on Adult Vaccination was carried out in a joint effort of the National Institute of Geriatrics, bringing together a group of specialists who worked on three central objectives: establishing vaccination guidelines throughout the life course, with emphasis on new vaccines; defining priority groups according to their risk factors; and contributing to the effort to promote healthy aging.

Previous studies with perfluorooctanesulfonic acid (PFOS) and perfluorooctanoic acid (PFOA) indicate that they act as endocrine disruptors, in addition to inducing alterations and damaging reproductive health; however, the biological mechanisms by which these disorders are produced are not yet understood. The aim of this study was to analyze the effect of PFOS and PFOA on in vitro steroidogenic secretion in porcine theca and granulosa cells, with or without gonadotropic stimulation. Granulosa and theca cells were isolated and cultured. Cell nature was performed by immunocytochemistry. PFOS and PFOA effect on steroid secretion was analyzed by chemiluminescence. In the present study, alterations in steroidogenic secretion were found when administering PFOS (0.12, 1.2, 12, 120 or 240 μM) or PFOA (0.012, 0.12, 1.2, 12 or 24 μM) to theca and granulosa cells. When theca and granulosa cells were stimulated with 500 ng/mL LH or 500 ng/mL FHS, respectively and immediately followed with 1.2 μM of PFOS or PFOA, the perfluorinated compounds inhibited the secretion of steroid hormones in both stimulated cell types. The results indicate that PFOS and PFOA act on steroidogenic ovarian cells as endocrine disruptors, which could affect the dependent functions of sexual steroids.

Summary Noncommunicable diseases (NCDs) are the leading causes of death globally, killing more people each year than all other causes combined. The principal factors that influence the emergence of NCDs are the combination of changes in urbanization, global trade markets, and longevity that has been occurring over decades in wealthy nations but are happening much faster in emerging countries. These changes come through the inability of developing countries to reestablish health and regulatory systems rapidly enough to keep pace with the changes. The consequences are devastating everywhere, but especially in vulnerable and disadvantaged populations. These people get sick sooner and die earlier than in wealthier societies. Usually, noncommunicable diseases are detected late, when patients need extensive and expensive hospital care for severe complications or acute events. Effective interventions are available, and evidence demonstrates their clear and measurable impact on a range of resource settings. Some of these interventions are continuing medical education as a response to the need to maintain postgraduate qualification and guarantee the quality of health care and promote the creation of care innovations that transform the primary health care system. The Carlos Slim Foundation established the CASALUD model to develop innovative public health strategies that transform primary care by improving the delivery of chronic disease management through the implementation of care innovations. This model includes electronic health education with PIEENSO ( http://pieenso.org/ ), an educational health platform established to contribute to the generation of sustained health initiatives that aim to have a profound impact on the people in Mexico and Latin America. PIEENSO provides health professionals academic tools to achieve meaningful learning and strengthening of skills in relevant public health issues. This model is among the first that includes education for physicians and other primary health care professionals, and focuses on CKD detection and prevention as a part of a comprehensive course on chronic noncommunicable diseases.

This study was designed to determine whether calreticulin (CRT), a chaperone protein, is present inin vitro-matured (IVM) pig oocytes and to study its potential role in the block to polyspermy. Western blot analysis, using an anti-CRT antibody, of oocyte lysate showed an immunoreactive band of ∼60 kDa. Simultaneous labeling of IVM oocytes with anti-CRT antibody and peanut agglutinin lectin (PNA lectin, a porcine cortical granules (CG)-specific binding lectin) revealed localization of CRT in the subplasmalemmal region with a 27.7% colocalization with PNA staining. After IVF, PNA labeling was not observed and anti-CRT labeling decreased significantly in zygotes and disappeared in two-cell embryos. Western blot analysis of oocyte exudate obtained from zona pellucida (ZP)-free oocytes activated with calcium ionophore confirmed the presence of a band that reacted with an anti-CRT antibody. Anti-CRT antibody and PNA labeling were not observed in activated oocytes despite being detectable in non-activated oocytes. The presence of CRT in vesicles located under the oolemma was demonstrated using immunogold cytochemistry at the ultrastructural level. To study the role of CRT in fertilization, ZP-enclosed and ZP-free oocytes were incubated with exogenous CRT and then inseminated. Whereas ZP-free oocytes showed fewer penetrating sperm and lower polyspermy rates than untreated oocytes, the opposite effect was observed in ZP-enclosed oocytes. In conclusion, CRT is confined to subplasmalemmal vesicles partially overlapping with CG contents. Its exocytosis after the oocyte activation seems to participate in the membrane block to polyspermy in pigs but is not involved in the ZP block.

The aim of this study was to determine the effect of Tai Chi on biological markers of oxidative stress in saliva and its relationship with periodontal disease (PD) in older adults. We carried out a quasi-experimental study with a sample of 71 sedentary volunteers with PD who were divided into a control group of 34 subjects and an experimental group of 37 subjects who performed Tai Chi 5 days a week for a period of 6 months. PD status was characterized using the Periodontal Disease Index (PDI). Superoxide dismutase (SOD), total antioxidant status (TAS), and TBARS levels of both groups were measured by spectrophotometric methods. In addition, inflammation markers (TNF-α, IL-1β, IL-6, IL-8, and IL-10) were measured by flow cytometry. We found a statistically significant increase in SOD activity (P<0.001) and TAS concentration (P<0.05), whereas levels of IL-1βwere significantly lower (P<0.01). Likewise, a statistically significant decrease in the PDI (P<0.05) was observed in subjects who performed Tai Chi during a period of 6 months. Our findings suggest that the practice of Tai Chi has both antioxidant and anti-inflammatory effects that are linked to the improvement of PD in older adults.

The aim of this study was to evaluate the effect of continuous exposure to sublethal ammonia concentrations upon the characteristics of the locomotor activity rhythms in Uca princeps, exposed to artificial tides and light cycles. Adult male crabs were used for standardized 24 h ammonia acute toxicity tests. Sublethal ammonia concentrations were selected considering the results obtained in the acute bioassays as well as the unusual high ammonia concentrations registered in their natural habitat. Three independent groups of eight crabs each were exposed during 15–20 days to 0.125, 3.5, and 18 mg N-TA/L in artificial sea water. Locomotor activity was recorded individually by means of infrared light crossings. Artificially created tidal and light–dark cycles were used along the experiments. The 24 h-LC50 was 186.02 mg N-TA/L. No mortality was registered in the crabs exposed to the sublethal ammonia concentrations. In control groups, organisms showed tidal activity onset mainly in low tide (ebb) and no respons...

Several coastal lagoons and estuaries in Mexico receive untreated domestic and industrial discharges which contain complex mixtures of contaminants. In order to assess the effects of chemical contamination, we used the White mullet (Mugil curema) as biological indicator. We worked in two estuaries located in Northwest Mexico: Urias (highly polluted) and Teacapan (less polluted, therefore used as reference site). We measured several endpoints at different levels of biological organization: vitellogenin transcription in males as biomarker of estrogenic contamination, as well as reproductive, morphological (deformities), morphometric, and meristic parameters. Total RNA was isolated from the liver, and a partial sequence of the mullet vitellogenin gene was obtained; gene expression was analyzed by quantitative PCR. At the same time, gonad samples were analyzed by histologic techniques to determine sex ratios and the reproductive cycle stage. The reproductive season was detected from February to June in both sites, but the gonadosomatic index was consistently higher in Teacapan. Sex ratios were female-biased in both estuaries, and one intersex gonad and several malformations were found in fish from Urias. Vitellogenin gene transcription in males was detected in both sites, although gene expression was slightly higher in Urias. The results obtained in this study indicate that biological effects of contamination are evident in fish, environmental estrogens may be present in both estuaries, and the white mullet is useful as biological indicator to identify and characterize environmental stressors in tropical coastal ecosystems.

The genus Bathypterois (tripod fish) comprises 19 species of deep-sea fishes distributed worldwide. The biology and distribution of the species of this genus are relatively poorly known throughout the Eastern Central Pacific (ECP). This work aims to update the geographic and bathymetric distribution of species of Bathypterois throughout the ECP. To describe the influence of depth, temperature and dissolved oxygen concentrations (DO) on latitudinal and depth distribution of members of the genus throughout the Mexican Pacific, 51 specimens of B.atricolor (12 trawls) and 112 of B. ventralis (18 trawls), collected during TALUD project, were analyzed. Bathypterois atricolor had broader environmental niches (depth: 0.47, temperature: 0.46, DO: 0.39), and inhabits deeper and colder sites (Electivity positives: > 1 000 m, < 3.2 °C, 1.0-1.62 mL/L) than B. ventralis, which had narrow environmental niches (depth: 0.34, temperature: 0.32, DO: 0.28), with high affinity to warmer sites and lower DO concentrations (Electivity positives: 778-1 400 m, 3.3-5.8 °C, < 1.0 mL/L). Both species used different environmental strata (niche overlap; depth: 0.34, temperature: 0.32, DO: 0.28). The adaptation of B. atricolor to live in a broad range of DO concentrations explains its almost circumglobal distribution, while B. ventralis lives almost permanently in the Oxygen Minimum Zone (< 0.05 mL/L), so the DO could be a limiting factor, reducing its potential geographic and bathymetric distributions. None of these species had been previously recorded in the central Gulf of California, and B. ventralis had not been recorded along the West coast of Baja California. This study confirms that both species have a continuous distribution within the ECP, our new records extended the known northernmost range for B. atricolor by 886 km (from 22°46’ N to 30°45’ N) and for B. ventralis by 645 km (from 21°18’ N to 27°07’ N). Rev. Biol. Trop. 65 (1): 89-101. Epub 2017 March 01. ResumenEl género Bathypterois (peces trípode) está conformado por 19 especies de peces de profundidad, distribuidas alrededor del mundo. La biología y distribución de las especies de este género son relativamente poco conocidas a lo largo del Pacífico Centro Oriental (POT). Este trabajo tiene el objetivo de actualizar la distribución geográfica y batimétrica de las especies del género Bathypterois a lo largo del POT. Para describir la influencia de la profundidad, temperatura y concentraciones de oxígeno disuelto (OD) sobre la distribución latitudinal y batimétrica de miembros del género Bathypterois en el Pacífico Mexicano, se analizaron 51 especímenes de B.atricolor (12 arrastres) y 112 B. ventralis (18 arrastres), recolectados durante el proyecto TALUD. Bathypterois atricolor presentó nichos ambientales más amplios (profundidad: 0.47, temperatura: 0.46, OD: 0.39) y habita sitios más profundos y fríos (Electividad positiva: > 1 000 m, < 3.2 °C, 1.0 - 1.62 mL/L) en comparación con B. ventralis, que tiene nichos ambientales reducidos (profundidad: 0.34, temperatura: 0.32, OD: 0.28), con alta afinidad por sitios más cálidos y bajas concentraciones de OD (Electividad positiva: 778-1 400 m, 3.3-5.8 °C, < 1.0 mL/L). Estas especies utilizaron distintos estratos ambientales (traslape de nicho; profundidad: 0.34, temperatura: 0.32, OD: 0.28). La adaptación de B. atricolor para vivir en un amplio intervalo de concentraciones de OD explica su distribución casi circunglobal, mientras que B. ventralis vive casi permanentemente en la Zona de Mínimo Oxígeno (< 0.05 mL/L), por lo que el OD puede ser un factor limitante, reduciendo su posible distribución geográfica y batimétrica. Ninguna de estas especies había sido registrada anteriormente en el centro del Golfo de California, así como B. ventralis no había sido reportada a lo largo de la costa occidental de Baja California. Este estudio confirma que ambas especies tienen una distribución continua dentro del POT, nuestros nuevos registros extienden la distribución más norteña conocida para B. atricolor en 886 km (de 22°46’ N a 30°45’ N) y en 645 km para B. ventralis (de 21°18’ N a 27°07’ N).

Malathion is one of the most commonly used insecticides. Recent findings have demonstrated that it induces oxidative stress in somatic cells, but there are not enough studies that have demonstrated this effect in germ cells. Malathion impairs porcine oocyte viability and maturation, but studies have not shown how oxidative stress damages maturation and which biochemical mechanisms are affected in this process in cumulus-oocyte complexes (COCs). The aims of the present study were to determine the amount of oxidative stress produced by malathion in porcine COCs matured in vitro, to define how biochemical mechanisms affect this process, and determine whether trolox can attenuate oxidative damage. Sublethal concentrations 0, 750, and 1000 µM were used to evaluate antioxidant enzyme expressions, reactive oxygen species (ROS production), protein oxidation, and lipid peroxidation, among other oxidation products. COCs viability and oocyte maturation decreased in a concentration-dependent manner. Malathion increased Cu, Zn superoxide dismutase (SOD1), glutathione-S-transferase (GST), and glucose 6 phosphate dehydrogenase (G6PD) protein level and decreased glutathione peroxidase (GSH-Px) and catalase (CAT) protein level. Species reactives of oxygen (ROS), protein oxidation and Thiobarbituric acid reactive substances (TBARS) levels increased in COCs exposed to the insecticide, but when COCs were pre-treated with the trolox (50 µM) 30 min before and during malathion exposure, these parameters decreased down to control levels. This study showed that malathion has a detrimental effect on COCs during in vitro maturation, inducing oxidative stress, while trolox attenuated malathion toxicity by decreasing oxidative damage.

Chagas disease is a leading cause of heart disease affecting approximately 10 million people in Latin America and elsewhere worldwide. The two major drugs available for the treatment of Chagas disease have limited efficacy in Trypanosoma cruzi-infected adults with indeterminate (patients who have seroconverted but do not yet show signs or symptoms) and determinate (patients who have both seroconverted and have clinical disease) status; they require prolonged treatment courses and are poorly tolerated and expensive. As an alternative to chemotherapy, an injectable therapeutic Chagas disease vaccine is under development to prevent or delay Chagasic cardiomyopathy in patients with indeterminate or determinate status. The bivalent vaccine will be comprised of two recombinant T. cruzi antigens, Tc24 and TSA-1, formulated on alum together with the Toll-like receptor 4 agonist, E6020. Proof-of-concept for the efficacy of these antigens was obtained in preclinical testing at the Autonomous University of Yucatan. Here the authors discuss the potential for a therapeutic Chagas vaccine as well as the progress made towards such a vaccine, and the authors articulate a roadmap for the development of the vaccine as planned by the nonprofit Sabin Vaccine Institute Product Development Partnership and Texas Children’s Hospital Center for Vaccine Development in collaboration with an international consortium of academic and industrial partners in Mexico, Germany, Japan, and the USA.

SummaryOogenesis is a highly complex process that requires the exquisite temporal and spatial regulation of gene expression at multiple levels. Skin–embryo–brain–oocyte homeobox (Sebox) gene encodes a transcription factor that is highly expressed in germinal vesicle stage oocytes and that plays an essential role in early embryogenesis at the 2-cell stage in the mouse. AsSeboxis also expressed in mouse fetal ovaries, the aim of the present study was to study its role during the early oogenesisin vitro. Expression ofSeboxwas low in 15.5 to 17.5 days post coitum (dpc) ovaries, showed a peak at 18.5 dpc and then its expression decreased dramatically in newborn ovaries.Seboxexpression was efficiently knocked down (>80%) in fetal mouse ovary explants in culture using RNAi technology. When fetal ovary explants were transfected withSebox-specific RNAi, the number of oocytes at germinal vesicle stage and showing a diameter of 40–70 μm was decreased significantly to 75% after 7 days of culture relative to the negative control, and to 22.4% after 10 days of culture, thus indicating thatSeboxplays an important role in the early oogenesis in mice.

Dengue infection is a significant and escalating public health problem in Latin America. Its re-emergence and subsequent rise in the region over the past 50 years has largely been caused by a combination of a lack of political will, the radical growth of urban populations, migration flow and insufficient financial resources. Its increased incidence has been compounded by climate change, poor sanitation and extreme poverty, which lead to more breeding sites of the mosquito vector Aedes aegypti. In order to control dengue effectively, an integrated approach incorporating vector management and environmental and social solutions is required. To achieve success, these programmes require commitment and responses at both national and community level. The development of a vaccine is a vital tool in the fight against dengue. For successful introduction, those implementing vaccination need to be educated on the value of such a strategy. Effective political leadership, innovative financial mechanisms and co-operation across all disciplines, sectors and national borders are essential to eradication of the disease.

We previously identified 62, 39, 27 and 7 kDa porcine sperm plasma membrane proteins that demonstrated a predominant affinity for the porcine oocyte plasma membrane by Western ligand blotting. The current experiments were designed to further investigate the potential roles of these molecules in sperm-oocyte plasma membrane interaction. Abilities of these proteins to bind to the oocyte plasma membrane and to inhibit sperm-oocyte interaction were evaluated. Plasma membrane was isolated primarily from the head of ejaculated porcine sperm by nitrogen cavitation and density gradient centrifugation. Fractions containing the 62, 39, 27 and 7 kDa proteins were electroeluted from one dimensional SDS polyacrylamide gels, dialysed and proteins biotinylated. Following incubation with zona-free porcine oocytes, bound protein was visualized with 20 μg TRITC-avidin/ml using confocal microscopy. Fractions of the dialysed, electroeluted proteins were added to porcine in vitro fertilization assays. The 62, 39, 27 and 7 kDa proteins all demonstrated binding to the oocyte plasma membrane in contrast to a biotinylated control protein. Addition of unlabelled sperm plasma membrane proteins to the biotinylated protein visibly reduced binding. Addition of each of these protein fractions to in vitro fertilization assays reduced sperm interaction with the porcine oocyte plasma membrane in a concentration-dependent manner. Binding of these sperm plasma membrane proteins to the oocyte plasma membrane and inhibition of fertilization are consistent with these proteins being involved in sperm-oocyte plasma membrane interaction.

Follicular apoptosis in the tropical mussel Mytella strigata was assessed in three coastal lagoons located in the southern Gulf of California, Mexico. Mussels were collected from three coastal lagoons associated with different scenarios of anthropogenic stress during one year. The gonad of each mussel was dissected, weighed, and sampled for histology and apoptosis analysis by TUNEL labeling. Two apoptotic indices were used: the apoptotic index of cells (AIC) based on the number of follicular cells in apoptosis in one thousand cells counted per gonad, and the apoptotic index of follicles (AIF) based on the number of follicular cells per follicle per gonad. Both indices showed high association with each other for all developmental stages, although AIF seemed to better discriminate among sites. Higher AIF and AIC were observed at the Urias Estuary (1.6 and 1.5 respectively) ranked as highly polluted, followed by Ensenada del Pabellon (0.82 and 0.95 respectively), ranked as moderately polluted, and the Teacapan Estuary (0.57 and 0.76 respectively) ranked as slightly polluted. Our data indicate that the apoptotic index in tropical mussels could be a useful indicator of environmental stress in coastal ecosystems; however, the ecological relevance of follicular apoptosis in polluted environments needs further investigation.

In an in vitro model using HEp-2 cells treated with purified plasmid-encoded toxin (Pet), we have identified morphological changes characterized by cell rounding and detachment after toxin internalization; these changes progress to cell death. However, these effects have not yet been shown to occur during the infection of epithelial cells by enteroaggregative Escherichia coli (EAEC). Here, we show that the secretion of Pet by EAEC is regulated at the transcriptional level, since secretion was inhibited in eukaryotic cell culture medium, although Pet was efficiently secreted in the same medium supplemented with tryptone. Inefficient secretion of Pet by EAEC in DMEM prevented cell detachment, whereas efficient Pet secretion in DMEM/tryptone increased cell detachment in a HEp-2 cell adherence assay. Interestingly, Pet toxin was efficiently delivered to epithelial cells, since it was internalized into epithelial cells infected with EAEC at similar concentrations to those obtained by using 37 μg ml−1 purified Pet protein. Additionally, Pet was not internalized when the epithelial cells were infected with a pet clone, HB101(pCEFN1), unlike the wild-type strain, which has a high adherence capability. There is a correlation between Pet secretion by EAEC, the internalization of Pet into epithelial cells, cell detachment and cell death in EAEC-infected cells. The ratio between live and dead cells decreased in cells treated with wild-type EAEC in comparison with cells treated with an isogenic mutant in the pet gene, whereas the effects were restored by complementing the mutant with the pet gene. All these data indicate that Pet is an important virulence factor in the pathogenesis of EAEC infection.

Atrazine is a herbicide of the chloro-s-triazine family. It inhibits photosynthesis in plants and is an endocrine disruptor, but its effects on human health are controversial. Fenoxaprop-ethyl, an aryloxy phenoxyalkanoic acid herbicide, inhibits the biosynthesis of fatty acids and provokes depolarization of membranes. The aim of this study is to evaluate the in vitro effects of both herbicides on capacitation, spontaneous acrosome reaction (SAR) and progesterone-induced acrosome reaction (PIAR) in boar sperm. Sperm capacitation is done in TALP-HEPES media for 4 hours. Capacitation and SAR are evaluated immediately; PIAR, 30 minutes later. LC50 for fenoxaprop-ethyl is 60 micromolar [corrected] and 40 micromolar [corrected] for atrazine. Fenoxaprop-ethyl induces capacitation at 60 micromolar [corrected] and SAR at all concentrations, also increases significantly PIAR. Atrazine decreased capacitation whereas increase significantly SAR and PIAR at all concentrations. It seems that fenoxaprop-ethyl and atrazine accelerate the capacitation and the acrosomal reaction, possibly via plasma membrane destabilization.

Concurrent infections provoked by the sea lice Lepeophtheirus simplex and the monogenean Neobenedenia sp. on bullseye puffer fish Sphoeroides annulatus, reared in tanks, have been related to skin pathology and mortality. Freshwater treatment is widely used to treat two parasite groups, but the results differ depending on the parasite species, life stage and exposure time. The present study describes the effectiveness of freshwater treatments to remove both parasites. Sea lice nauplii and free swimming copepodids, reared in the laboratory, were exposed to freshwater treatments for periods from 30 s to 16 min. Fish experimentally infected with sea lice or monogeneans were exposed to freshwater for 15 to 90 min and 10 to 60 min, respectively. All sea lice nauplii and copepodids were killed by freshwater dips at 2.5 and 14 min, respectively. For pre-adult and adult sea lice, 30 and 60 min of freshwater exposure significantly reduced up to 95 and 99% respectively. However, most freshwater exposures did not reduce chalimus stages more than 10%. Immature and adult populations of monogeneans showed a significant reduction of 95% at 20 min of exposure. These results indicate that, if used properly, freshwater baths constitute an efficient alternative to control these mixed ectoparasite infections on bullseye puffer fish reared in tanks.

Malathion and diazinon are two of the most commonly used organophosphorous (OP) agrochemicals. Several studies show that these pesticides exert several effects on mammalian spermatogenesis. Nevertheless, there are no studies concerning their effects on oogenesis. The objective of this study was to evaluate the effects of these insecticides on the viability of in vitro cultured mouse oocytes during the early oogenesis and to get a further understanding of the molecular mechanisms by which OP insecticides act and affect germinal cells. Oocytes were cultured from fetal ovaries for 10 days, when most oocytes had reached the diplotene stage (germinal vesicle stage). Cultures were exposed to different concentrations of malathion or diazinon for 24 h, and the effect on oocyte viability was assessed. Gene expression in oocytes exposed to the insecticides was analyzed by generating cDNA libraries and performing differential screenings. Results show a significant decrease in oocytes survival after 24-h exposure to 250 μM malathion or 900 nM diazinon, and the effect of these insecticides on the regulation of genes encoding proteins involved in transcription (BP75), translation (ribosomal protein S5), and mitochondrial function (cytochrome oxidase subunits I and III), providing evidence for OP insecticides as toxicants for mammals oocytes during the early oogenesis. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2008.

Fish at the top of the food chain bioaccumulate and biomagnify toxic metals including mercury (Hg), at a higher rate than nonpredatory fish. However, although some top predator fish species are important in the human diet, the risks for consumers in Mexico are difficult to evaluate due to the scarce baseline information available. In the present article, data on the total mercury (THg) concentrations found in edible tissues of different species of sharks, rays, large pelagic fishes and groupers from the Gulf of California are presented and compared with national and international health guidelines. During 2003 and 2004, 73 shark dorsal tissue samples, 52 ray samples, 66 large pelagic fish samples and 16 grouper samples were collected at different sites along the Gulf of California. Samples were digested in a microwave system and analyzed by cold vapor atomic absorption spectrophotometry. Thirty samples (14%) showed concentrations of Hg above the national statutory limit of 1 mg kg−1, from which 25 samples...

Ancyrocephaline monogeneans such as Haliotrema sp. and Euryhaliotrema sp. are common parasites infecting gills on the red snapper, Lutjanus guttatus , an important potential fish species for aquaculture in Mexico. The effectiveness of different treatments against these monogeneans using in vitro and in vivo assessments on the viability of eggs and adults was evaluated. Eggs from both monogenean species were in vitro exposed to sodium hypochlorite, drying, formalin, and freshwater for 3 h, and returned to seawater to determine post-treatment hatching. Adults of both parasites were also in vitro exposed to formalin, freshwater, and caprylic acid for periods ranging from 10 to 60 min using seawater as control. In vivo tests were carried out with freshwater, formalin, and praziquantel (active ingredient of Drontal™ Plus and Vermiplex™ Plus) on snappers infected with both parasites. After 3 h exposure, egg hatching was completely suppressed at 120 mg l − 1 of available chlorine, while drying, formalin (83 mg l − 1 ) and freshwater showed eggs viabilities of 10, 23, and 10% respectively. In vitro freshwater exposure significantly reduced the mean intensity of ancyrocephaline adults (99% at 30 min), followed by formalin at 51 mg l − 1 (72% at 60 min) and caprylic acid at 334 mg l − 1 (5% at 30 min). In vivo freshwater baths for 30 min removed 100% of ancyrocephalines, although this method was stressful for the fish, while formalin (60 min at 51 mg l − 1 ), Drontal™ Plus (12-h at 4.5 mg l − 1 ) and Vermiplex™ Plus (24-h at 3.5–4.5 mg l − 1 ) removed 72, 100 and 100% respectively. This study indicates that praziquantel, freshwater, and formalin baths were effective against ancyrocephaline adults from red snapper, and sodium hypochlorite would be useful to prevent horizontal infection with eggs on fish-rearing facilities.

Mercury (Hg) and cadmium (Cd) concentrations in eggs of 8 seabird species inhabiting 5 coastal ecosystems in Sinaloa, México were determined during 2 breeding seasons (2012 and 2013): blue-footed booby (Sula nebouxii), brown booby (Sula leucogaster), double-crested cormorant (Phalacrocorax auritus), magnificent frigatebird (Fregata magnificens), brown pelican (Pelecanus occidentalis), royal tern (Thalasseus maximus), laughing gull (Leucophaeus atricilla), and Heermann's gull (Larus heermanni). The interspecific differences found in the concentrations of both metals were attributed to the diet and foraging ecology of the species. The highest Hg concentrations were detected in piscivorous species (brown pelican, 0.42 µg/g; brown booby, 0.31 µg/g; blue-footed booby, 0.26 µg/g; and double-crested cormorant, 0.23 µg/g); whereas species with more varied diets presented the highest Cd concentrations (Heermann's gull, 0.31 µg/g; laughing gull, 0.27 µg/g; and magnificent frigatebird, 0.27 µg/g). Cadmium concentrations were significantly greater in 2013 than 2012 for most species, and brown pelican and laughing gull also had higher Hg concentrations in 2013 in Santa María Bay, suggesting a relationship as a result of the changes either in oceanographic conditions or in continental runoff. Mercury concentrations in brown pelican and Cd concentrations in Heermann's gull and laughing gull were above threshold levels for adverse effects on reproduction and survival. Environ Toxicol Chem 2016;35:2330-2338. © 2016 SETAC.

This study was designed to evaluate the capacity of vitrified-warmed porcine immature oocytes to mature and to be fertilized using in vitro fertilization or intracytoplasmic sperm injection, and to determine the subsequent embryo development. Immature oocytes were vitrified using ethylene glycol and dimethylsulphoxide as cryoprotectants and the Cryolock method. After warming oocytes were cultured 44 h for maturation. Oocytes were randomly distributed in three treatment groups and subjected to in vitro fertilization (Experiment 1) or intracytoplasmic sperm injection (Experiment 2) procedures. The results indicate that the embryo development was higher in denuded oocytes co-cultured with granulosa cells (NkO-CC group) fertilized by in vitro fertilization or intracytoplasmic sperm injection compared to cumulus-cell oocyte complexes (COCs group), showing no significant differences with control. Vitrified denuded oocytes matured with a co-culture system NkO-CC group, displayed higher cleavage rate and blastocyst production than vitrified COCs group. Blastocysts were successfully obtained after IVF and ICSI procedures; however, the development to the blastocyst stage was better after IVF. These results show that the vitrification-warming media, the employment of a granulosa cell co-culture system and the Cryolock method during vitrification, increased the nuclear and cytoplasmic maturation of vitrified porcine immature oocytes. Further experiments are required to enhance porcine embryo production after vitrification.

Background Dengue imposes a substantial economic and disease burden in most tropical and subtropical countries. Dengue incidence and severity have dramatically increased in Mexico during the past decades. Having objective and comparable estimates of the economic burden of dengue is essential to inform health policy, increase disease awareness, and assess the impact of dengue prevention and control technologies. Methods and Findings We estimated the annual economic and disease burden of dengue in Mexico for the years 2010–2011. We merged multiple data sources, including a prospective cohort study; patient interviews and macro-costing from major hospitals; surveillance, budget, and health data from the Ministry of Health; WHO cost estimates; and available literature. We conducted a probabilistic sensitivity analysis using Monte Carlo simulations to derive 95% certainty levels (CL) for our estimates. Results suggest that Mexico had about 139,000 (95%CL: 128,000–253,000) symptomatic and 119 (95%CL: 75–171) fatal dengue episodes annually on average (2010–2011), compared to an average of 30,941 symptomatic and 59 fatal dengue episodes reported. The annual cost, including surveillance and vector control, was US$170 (95%CL: 151–292) million, or $1.56 (95%CL: 1.38–2.68) per capita, comparable to other countries in the region. Of this, $87 (95%CL: 87–209) million or $0.80 per capita (95%CL: 0.62–1.12) corresponds to illness. Annual disease burden averaged 65 (95%CL: 36–99) disability-adjusted life years (DALYs) per million population. Inclusion of long-term sequelae, co-morbidities, impact on tourism, and health system disruption during outbreaks would further increase estimated economic and disease burden. Conclusion With this study, Mexico joins Panama, Puerto Rico, Nicaragua, and Thailand as the only countries or areas worldwide with comprehensive (illness and preventive) empirical estimates of dengue burden. Burden varies annually; during an outbreak, dengue burden may be significantly higher than that of the pre-vaccine level of rotavirus diarrhea. In sum, Mexico’s potential economic benefits from dengue control would be substantial., Author Summary During the past decades, dengue fever has become the most common arthropod-borne viral disease, imposing a substantial economic and disease burden in most tropical and subtropical countries, including Mexico. Dengue incidence and severity have dramatically increased in Mexico, with transmission regularly reported in 28 of 32 states. Objective estimates of the burden of dengue are important to inform policy decisions and priorities. We merged multiple data sources to estimate (i) total episodes, (ii) costs per episode, (iii) surveillance and vector control costs, and (iv) disease burden (2010–2011). Results suggest that Mexico had about 139,000 symptomatic and 119 fatal dengue episodes per year on average. The annual cost, including surveillance and vector control, was about US$170 million, or $1.56 per capita, comparable to other countries in the Americas. Annual disease burden averaged 65 disability-adjusted life years per million population, with most of the years lost to disability corresponding to ambulatory episodes. The results show a substantial burden of dengue on the health care system and economy of Mexico. This quantification of the economic burden should help public health officials make informed decisions about current and promising new preventive and control measures to reduce dengue infections.

The recent decline in sperm concentration observed in men has developed over a short period of time, suggesting that it could be the result of environmental factors. The present study has evaluated the effects of insecticides Malathion and Diazinon, and herbicides Atrazine and Fenoxaprop-Ethyl on porcine sperm viability and motility patterns in vitro using the eosin-nigrosin staining and a computer-assisted semen analyzer (CASA), respectively. Malathion and Fenoxaprop-Ethyl exerted more deleterious effects than Diazinon and Atrazine. Progressive sperm motility was strongly affected whereas the effect on sperm viability was less pronounced. This suggests that a reduction of sperm motility is not necessarily the result of sperm death. Since sperm motility is dependent on energy metabolism the mechanism of action of these pesticides might be mediated at the level of the mitochondrion, producing a delay in motility and eventual cell death.

The relationship between parasites and environmental stress were studied in two tropical coastal lagoons of Northwest Mexico: Urias estuary (highly polluted) and Teacapan estuary (slightly polluted). Metazoan parasites were examined in 292 white mullet (Mugil curema) specimens collected bimonthly during a year from both systems. Haliotrema mugilinus, Metamicrocotyla macracantha, Ergasilus sp., Caligus sp., Holobomolochus sp., and Lernaeopodidae were found in gills, while Contracaecum sp. larvae III was found liver, hepatic portal vein and kidneys. Ecological indices were influenced by the slightly higher number of parasitic species in Urias compared to Teacapan, as well as the clear dominance of two species: Ergasilus sp. and Contracaecum sp. in both systems. In fact, Ergasilus sp. showed considerably higher abundance in Urias, possibly indicating that its success was a result of adverse conditions affecting the host, while Contracaecum sp showed higher abundances in Teacapan, suggesting that the environmental conditions occurring in Urias could have produced negative impacts on the nematode's infective potential.

A tropical marine bioassay was developed with juveniles of Pacific white shrimp, Litopenaeus vannamei, in order to test the acute and sublethal toxicity of Tilt, which is the commercial formulation of the fungicide propiconazole. A 10-d acute toxicity and a 32-d sublethal test were performed. A median lethal concentration (LC50) was determined for different exposure times, resulting in a 24-h LC50 of 1167 (1101-1386) microg/L (concentration based on active ingredient propiconazole), and reaching a threshold LC50 (72-h) at 1043 (1018-1068) microg/L. The sublethal exposure test was performed with propiconazole concentrations ranging from 367 to 825 microg/L. Animals in most treatments showed a significant increase in intermoult duration compared to those in the control treatment, although this did not seem to be concentration-dependent. However, the shrimps exposed to the sublethal concentrations of this fungicide showed morphological deformities, with a significant positive relationship between concentration and deformities of the rostrum, paraeopods, and uropods. Altogether, the results presented indicate the potential effects of Tilt on shrimp, particularly under long-term exposures, although these were found to occur at concentrations exceeding those reported in natural environments.

En 1992 apareció en el sureste asi��tico un nuevo tipo de virus de la influenza, el cual ha ocasionado hasta la fecha más de 120 casos y un poco más de 60 defunciones en humanos en Camboya, Vietnam, Indonesia y Tailandia. Esta situación es considerada por los expertos como la probable génesis de una nueva pandemia de influenza, lo que podría traer graves consecuencias para la salud de la población, así como para la economía y el comercio mundial. Por lo anterior, la Organización Mundial de la Salud (OMS) ha instado a los países miembros a desarrollar planes de preparación y respuesta para hacer frente a esta eventualidad. En el marco del Comité Nacional para la Seguridad en Salud, México ha diseñado el Plan Nacional de Preparación y Respuesta ante una Pandemia de Influenza con objeto de proteger a la población mediante acciones efectivas y oportunas. El Plan utiliza una escala de riesgo y define cinco líneas de acción: Coordinación, Vigilancia Epidemiológica, Atención Médica, Difusión y Movilización Social, y Reserva Estratégica. Si bien es imposible predecir cuándo se presentará la próxima pandemia y su impacto, es fundamental que las autoridades de salud nacionales, estatales y locales establezcan los mecanismos para poner en marcha los componentes del Plan en forma oportuna y garantizar con ello la salud de la población en caso de influenza pandémica.

Sublethal effects of foodborne exposure to methyl parathion (0.62 and 1.31 microg methyl parathion*g(-1) dry weight of food) on juveniles of Litopenaeus vannamei using integrated biochemical (acetylcholinesterase (AChE) and ATPases) and physiological (feeding rate (FR), egestion rate (ER), and hepatosomatic index (HI)) biomarkers were evaluated. The HI was significantly higher in controls than in pesticide treatments. The FR was significantly lower in controls than in pesticide treatments while no significant differences were detected in the ER. AChE activity was significantly higher in controls than in pesticide treatments (control = 0.11 +/- 0.02; solvent control = 0.11 +/- 0.03; 0.62 = 0.07 +/- 0.01; 1.31 = 0.08 +/- 0.02 microM*min(-1)*mgprotein(-1)). The total-ATPase activity was significantly lower in controls than in pesticide treatments (control=77.90+/-12.41; solvent control = 83.69 +/- 22.05; 0.62 = 110.03 +/- 22.17; 1.31 = 121.54 +/- 19.84 microM P(i)*h(-1)*mgprotein(-1)). The Mg(2+)-ATPase activity was significantly higher in treatments than in controls (control = 65.14+/-10.76; solvent control = 75.12 +/- 21.10; 0.62 = 100.53 +/- 20.97; 1.31 = 108.94 +/- 17.26 microM P(i)*h(-1)*mgprotein(-1)). Finally, the results obtained for the Na(+)/K(+)-ATPase activity were significantly higher in control and in 1.31 than in solvent control and in 0.62 (control = 14.06+/-2.63; solvent control=7.30 +/- 4.13; 0.62 = 7.60 +/- 3.81; 1.31 = 13.42 +/- 2.88 microM P(i)*h(-1)*mgprotein(-1)). The results in this study showed that pulse exposures to methyl parathion via food could elicit measurable effects on the marine shrimp L. vannamei, indicating that foodborne exposure can be a reliable toxicological procedure and, if combined with pulse exposures, could also simulate more realistic exposure scenarios.

The causes of disease in cultured shrimp are difficult to ascertain but there is evidence that disease is correlated with environmental factors. Crustaceans are particularly sensitive to insecticides due to their close phylogenetic relationship with insects. The objective of this study was to investigate whether there was an increased susceptibility of the shrimp Litopenaeus vannamei to Vibrio parahaemolyticus, when exposed to methyl parathion. The outline of the study was the following: An LC50 96 h was determined to methyl parathion orally offered to juvenile shrimp. Further experiments were carried out in order to determine a concentration that affected the shrimp (verified by measuring the acethylcholinesterase activity) while producing minimal mortalities. This sublethal concentration was used in a susceptibility experiment where methyl parathion was offered to shrimp which were later injected with V. parahaemolyticus in a dose expected to kill less than 15%. Probit analysis estimated a 96 h median lethal concentration (LC50) of 1.56 l gg 1 . Mortality and AChE activity showed a concentration–response relationship in the exposure treatments. The median inhibitory concentration (IC50) determined was 0.029 l gg 1 . These results suggested that a concentration of 0.1 l gg 1 was appropriate for the bacteria–pesticide interaction test as it was able to elicit 11.1% mortality after 10 days of exposure, while producing an AChE inhibition of 57.12%. Cumulative mortalities were significantly increased (P < 0.01) in the treatment that combined exposure to methyl parathion and V. parahaemolyticus (35.19%) in comparison with methyl parathion or V. parahaemolyticus alone (9.26% and 7.41%, respectively). � 2005 Elsevier Ltd. All rights reserved.

We have purified and characterized pig and bovine milk lactadherins. Studies by circular dichroism spectroscopy indicate that the two proteins present a similar folding pattern. Results have been discussed in terms of their affinity for pig zona pellucida in order to use these proteins as analogs of pig sperm lactadherin in gamete studies.

Artificial insemination (AI) has been used for avian reproduction due to the discovery of cryoprotectants extending its usefulness both in production of domestic fowl and conservation of wild species. The goal of this study was to assess the effect on domestic and wild fowl pooled semen and individual ejaculate cryopreservation with dimethyl sulfoxide (DMSO) and polyvinylpyrrolidone (PVP). Twenty ejaculates and twenty samples of pooled semen of roosters, pheasants and hawks were frozen in media containing DMSO or PVP. DMSO and PVP cryopreservation are equally effective both for ejaculates and pooled semen. Even PVP is a good alternative since no significant difference was found when compared to DMSO. The fertilizing capacity of fresh and cryopreserved pooled semen was analyzed through AI of hens and female pheasants. Similar fertility rates using DMSO, PVP or frozen-thawed samples demonstrated that reproduction is possible through the use of cryopreserved semen. In the case of female pheasants, the same values were obtained with both cryopreserved and fresh semen.

Conventional cytogenetics (CC) can be used to identify chromosomal abnormalities that are predictors of treatment outcome in acute lymphoblastic leukemia (ALL). The detection of abnormalities in ALL is difficult because low mitotic index and poor-quality metaphases are obtained. Flow cytometry (FC) and fluorescence in situ hybridization (FISH) can be used to detect aneuploidy in any phase of the cell cycle, increasing the number of analyzable cells. The aim of this study was to develop a strategy combining these methods to improve the frequency of chromosome abnormality detection. One hundred children with newly diagnosed ALL were included. CC and DNA content analysis by FC were performed in all patients. The numerical abnormalities identified by both methods were compared and patients were classified as concordant or discordant. FISH was used to support aneuploidy results in discrepant cases using centromeric probes for the chromosomes most frequently involved in aneuploidy. CC and FC showed high concordance (86%). Fourteen cases were discrepant: nine showed hypodiploidy and low hyperdiploidy by cytogenetics and five showed high hyperdiploidy by FC. FISH confirmed aneuploidy in 12 cases in which it could be performed. High hyperdiploidy was the most common abnormality; the 31 cases showing this aneuploidy were identified by FC. The search for abnormalities must begin by measuring DNA content to detect this aneuploidy, which is useful to evaluate the patient's risk. However, it is important to screen for structural abnormalities by CC or molecular techniques. This strategy may detect chromosomal abnormalities, optimizing resources in laboratories where not all the screening methods are available.

First attempts at culturing Sphoeroides annulatus (bullseye puffer fish) resulted in recurrent parasitic infestations caused by the monogenea Heterobothrium ecuadori and the dinoflagellate Amyloodinium ocellatum, both of which are associated to mortality events in other cultured species. However, little information exists on the therapeutic levels to prevent and control such infections. The objective of the present study was to evaluate the safety of formalin to bullseye puffer fish and determine the efficacy of formalin to control parasitic infestations. The formalin median lethal concentration (LC50) for the bullseye puffer fish ranged from 1095 mg l−1 at 30 min and 972 mg l−1 at 60 min to 79 mg l−1 at 72 h. The formalin in vitro effective median concentration (EC50) against H. ecuadori was 225 mg l−1 at 30 min and 87 mg l−1 at 60 min. The resulting therapeutic index was 5 at 30 min and 11 at 60 min. Formalin was further tested in vivo against A. ocellatum. In comparison to the controls, 51 mg l−1 of formalin significantly reduced the number of parasites on the skin (97%) and gills (68%) of fish after 1 h of exposure, while 4 mg l−1 significantly reduced the numbers on the skin (66%) and gills (84%) after 7 h. The differences in toxicity between the bullseye puffer fish and specific parasites indicated that formalin was efficacious in the control of parasitic epizootics in the bullseye puffer fish culture.