Fertilizing ability of cryopreserved spermatozoa in the Persian sturgeon (Acipenser persicus) and stellate sturgeon (A. stellatus)

Motility of spermatozoa was studied on 12 and 7 specimens of Acipenser persicus and A. stellatus, respectively. The density measured to be 2.22±0.65x109ml-1 in A. persicus and 2.21±0.55x109ml-1 in A. stellatus. Semen samples were diluted with two extenders containing tris 118mM, sucrose 23.4mM, pH=8, egg yolk (20%), dimethyl sulfoxide (15%) and penicillin potassium (500IU/ml) and biociphus extender containing glycerol as a cryoprotectant at a ratio of 1:1 and then transferred to 0.5ml straws and frozen in a computer controlled low temperature apparatus and stored in liquid nitrogen for one week. To study fertilizing ability, the spermatozoa were then used to inseminate eggs after thawing. Mean sperm motility in fresh spermatozoa was 86.6% in A. persicus and 73.75% in A. stellatus which decreased to 32.2% (P<0.001) and 37.5% (P<0.001) in frozen spermatozoa, respectively. Also mean fertilization rate decreased from 90.4% to 30.7% in A. persicus and from 72% to 36.8% in A. stellatus

Gynogenesis in Persian sturgeon (Acipenser persicus) and beluga (Huso huso)

The objective of the present study was to determine the possible production of Persian sturgeon (Acipenser persicus) and Beluga (Huso huso) gynogen/triploids and also to determine the most appropriate type of thermal shock and the duration of induced shock after fertilization. Persian sturgeon and Beluga spawners were collected from Guilan's sturgeon catch stations and transported to the Shahid Beheshti sturgeon hatchery for artificial breeding and restocking programs. Ovulated eggs and sperms were collected based on common procedures in hatcheries. In order to separate the seminal fluids and dilute the milts, sperms were centrifuged at 6000 rpm for 20 min. and seminal fluids stored in refrigerator for further use. Sperm motility was investigated. In order to determine the best duration for radiation, the milt was diluted (1:9) with immobilizing solution. Samples of diluted milt were placed for UV irradiation (UV lamp model UVG-54, 254 nm, made by UVP America) for 0.5, 1, 1.5, 1.45, 2, to 5 min. The motility of radiated sperms and controls were examined under the light microscope and the motility curve was drawn. For application of thermal shock two types of heat shock (32, 34 and 37°C) and cold shock (0±1°C) were used for duration of 2.5 and 60 min respectively. Both thermal shock were applied at 12, 15, 18 min after fertilization. Four experimental groups were designed including; normal eggs as control group and sperms without UV thermal shock), gynogenesis (Sperm irradiated with UV and thermal shock were applied), triploid (thermal shock without radiation by UV on sperm) and haploid group (without thermal shock but using irradiated sperm for fertilization). Verification of the success of treatments was assessed using genetic analysis on sturgeon larvae and fingerlings. In triploids the total surface area, volume of cells and nucleus as well as chromosome number were determined. To identify a gynogenetic larva, microsatellite markers were used to analysis specific loci by using primers designed for lake sturgeon. The results were analyzed using SPSS, Excell software. To determine the significant levels between various parameters and comparison between controls and various treatments, one way of Analysis of Variance (ANOVA) was used. Whenever the significant level was observed to determine its level a Duncan test were examined. Results of present study showed that the best duration for UV radiation on sperms of Beluga was 105-110 seconds. Average fertilization rate for control Beluga was 51%, while in heat shock group it was 2-5 % and in cold shock it was 44.6%. There was a significant difference in fertilization rate in cold shock group compared to heat shock group (P<0.05), however no difference was observed between 32 and 34°C treatments. The average survival rate of larvae in control group was 51%, while in heat shock treatment (32 and 34°C) it was very low close to zero. However in cold shock treatment the results was better and hatching percentage of larvae was between 30 -35%. Triploid treatment showed better results than gynogenesis group. A minimum triploid larvae obtained from heat shock was zero but using cold shock, the maximum number of 170 specimen was harvested. There was no significant difference in the number of larvae obtained between 32 and 34° C treatments (P<0.05). Although some difference was observed on large and small axes, surface areas and volume of red blood cells but no significant differences were observed between control and triploid groups (P 0.05). In the meantime, the chromosome number in triploid beluga was (3N=177±3) as compared to diploid 2N= 118±3, which indicated an extra set of chromosome (n=60) in triploid fish. Totally 26.6% of investigated fish was triploids. Microsatellite molecular markers clearly differentiate gynogenetic fish on the bases of allele inheritance of male and female parents, and were proven that this technique can clearly identify allelic inheritance of parents to offspring. In Persian sturgeon in compare to beluga a different results were observed. Heat shock (37°C) not present any positive results therefore has no application in induce gynogenesis on this species, also no significant difference was observed between 32 and 34°C treatment. Cold shock showed better results, especially when duration of UV radiation was adjusted to 105 seconds. Molecular analysis using microsatellite marker positively proved the gynogenetic offspring by counting the allelic inheritance. However Persian sturgeon as a tetraploid species (2N=240) has its difficulty on scoring the banding patterns. We highly recommend disomic primers application for allelic inheritance on gynogene Persian sturgeon

Creation of cryopreservation bank of bony fish

In this study, 11 male of Caspian trout (Salmo truta caspius) (with mean length and weight 37/8 ± 5/3 cm and 523/3 ± 24/7 respectively) and 23 male of Caspian kutum (Rutilus frisii kutum) (with mean length and weight 36/1 ± 7/1 cm and 631/3 ± 21/6 g respectively) were evaluated. All the fish were good at the initial examination of sexual maturity. After sperm sampling, their quality were tested. In this step, the parameters such as motility, duration of mobility, density, pH and osmolality were measured. After this stage, the sperm samples of Caspian trout in the ratio 1: 3 were diluted with the aqueous solution containing compounds (0.3M Glucose, 10% Methanol, 10% egg yolk) and the freezing process was done manually and the sperm was frozen in liquid nitrogen. The sperm samples of Caspian kutums were diluted (ratio of 1: 3) with two soluble diluent containing compounds (350 mM glucose, 30 mM Tris and 4% Polyethylene glycol) and (350 mM glucose, 30 mM Tris and 2% Glycerol) and were frizzed automatically by Planner Kryo instrument and placed in liquid nitrogen. The sperm samples were thawed 1 to 3 months after the date of first freezing and their quality were assessed by measuring percent and timing motility. The results showed that the obtained semen volume of Caspian trout was more than Caspian kutum. Moreover, percentage of motile sperm, timing motility and sperm density of Caspian trout were higher than those of Caspian kutum but osmolality and pH of Caspian trout were lower than those of Caspian kutum. Over time, the percentage of sperm motility and mobility for both species declined compared with fresh samples. After thawing, percentage of motile sperm and timing motility of Caspian kutum were lower than those factors Caspian trout. The results showed that the sample of Caspian kutum sperm that were diluted by ethylene glycol after thawing and were immotile ll of them. However, the samples were diluted by glycerol, after thawing, were alive and motile. According to the results, it seems very important species differences that must be fully considered in the process of freezing sperm. The use of a single protocol would not be successful in cryopreservation because the reaction of sperm against to chemical agents is variable. Therefore, it is essential to get the right information to protect valuable Caspian fish by using cryopreservation. Further studies on the characteristics of each species, as well as the freezing process take appropriate diluent

The global, regional, and national burden of inflammatory bowel disease in 195 countries and territories, 1990�2017: a systematic analysis for the Global Burden of Disease Study 2017

Background: The burden of inflammatory bowel disease (IBD) is rising globally, with substantial variation in levels and trends of disease in different countries and regions. Understanding these geographical differences is crucial for formulating effective strategies for preventing and treating IBD. We report the prevalence, mortality, and overall burden of IBD in 195 countries and territories between 1990 and 2017, based on data from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2017. Methods: We modelled mortality due to IBD using a standard Cause of Death Ensemble model including data mainly from vital registrations. To estimate the non-fatal burden, we used data presented in primary studies, hospital discharges, and claims data, and used DisMod-MR 2.1, a Bayesian meta-regression tool, to ensure consistency between measures. Mortality, prevalence, years of life lost (YLLs) due to premature death, years lived with disability (YLDs), and disability-adjusted life-years (DALYs) were estimated. All of the estimates were reported as numbers and rates per 100 000 population, with 95 uncertainty intervals (UI). Findings: In 2017, there were 6·8 million (95 UI 6·4�7·3) cases of IBD globally. The age-standardised prevalence rate increased from 79·5 (75·9�83·5) per 100 000 population in 1990 to 84·3 (79·2�89·9) per 100 000 population in 2017. The age-standardised death rate decreased from 0·61 (0·55�0·69) per 100 000 population in 1990 to 0·51 (0·42�0·54) per 100 000 population in 2017. At the GBD regional level, the highest age-standardised prevalence rate in 2017 occurred in high-income North America (422·0 398·7�446·1 per 100 000) and the lowest age-standardised prevalence rates were observed in the Caribbean (6·7 6·3�7·2 per 100 000 population). High Socio-demographic Index (SDI) locations had the highest age-standardised prevalence rate, while low SDI regions had the lowest age-standardised prevalence rate. At the national level, the USA had the highest age-standardised prevalence rate (464·5 438·6�490·9 per 100 000 population), followed by the UK (449·6 420·6�481·6 per 100 000). Vanuatu had the highest age-standardised death rate in 2017 (1·8 0·8�3·2 per 100 000 population) and Singapore had the lowest (0·08 0·06�0·14 per 100 000 population). The total YLDs attributed to IBD almost doubled over the study period, from 0·56 million (0·39�0·77) in 1990 to 1·02 million (0·71�1·38) in 2017. The age-standardised rate of DALYs decreased from 26·5 (21·0�33·0) per 100 000 population in 1990 to 23·2 (19·1�27·8) per 100 000 population in 2017. Interpretation: The prevalence of IBD increased substantially in many regions from 1990 to 2017, which might pose a substantial social and economic burden on governments and health systems in the coming years. Our findings can be useful for policy makers developing strategies to tackle IBD, including the education of specialised personnel to address the burden of this complex disease. Funding: Bill & Melinda Gates Foundation. © 2020 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 licens

The global, regional, and national burden of inflammatory bowel disease in 195 countries and territories, 1990-2017 : a systematic analysis for the Global Burden of Disease Study 2017

Background The burden of inflammatory bowel disease (IBD) is rising globally, with substantial variation in levels and trends of disease in different countries and regions. Understanding these geographical differences is crucial for formulating effective strategies for preventing and treating IBD. We report the prevalence, mortality, and overall burden of IBD in 195 countries and territories between 1990 and 2017, based on data from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2017. Methods We modelled mortality due to IBD using a standard Cause of Death Ensemble model including data mainly from vital registrations. To estimate the non-fatal burden, we used data presented in primary studies, hospital discharges, and claims data, and used DisMod-MR 2.1, a Bayesian meta-regression tool, to ensure consistency between measures. Mortality, prevalence, years of life lost (YLLs) due to premature death, years lived with disability (YLDs), and disability-adjusted life-years (DALYs) were estimated. All of the estimates were reported as numbers and rates per 100 000 population, with 95% uncertainty intervals (UI). Findings In 2017, there were 6.8 million (95% UI 6.4-7.3) cases of IBD globally. The age-standardised prevalence rate increased from 79.5 (75.9-83.5) per 100 000 population in 1990 to 84.3 (79.2-89.9) per 100 000 population in 2017. The age-standardised death rate decreased from 0.61 (0.55-0.69) per 100 000 population in 1990 to 0.51 (0.42-0.54) per 100 000 population in 2017. At the GBD regional level, the highest age-standardised prevalence rate in 2017 occurred in high-income North America (422.0 [398.7-446.1] per 100 000) and the lowest age-standardised prevalence rates were observed in the Caribbean (6.7 [6.3-7.2] per 100 000 population). High Sociodemographic Index (SDI) locations had the highest age-standardised prevalence rate, while low SDI regions had the lowest age-standardised prevalence rate. At the national level, the USA had the highest age-standardised prevalence rate (464.5 [438.6-490.9] per 100 000 population), followed by the UK (449.6 [420.6-481.6] per 100 000). Vanuatu had the highest age-standardised death rate in 2017 (1.8 [0.8-3.2] per 100 000 population) and Singapore had the lowest (0.08 [0.06-0.14] per 100 000 population). The total YLDs attributed to IBD almost doubled over the study period, from 0.56 million (0.39-0.77) in 1990 to 1.02 million (0.71-1.38) in 2017. The age-standardised rate of DALYs decreased from 26.5 (21.0-33.0) per 100 000 population in 1990 to 23.2 (19.1-27.8) per 100 000 population in 2017. Interpretation The prevalence of IBD increased substantially in many regions from 1990 to 2017, which might pose a substantial social and economic burden on governments and health systems in the coming years. Our findings can be useful for policy makers developing strategies to tackle IBD, including the education of specialised personnel to address the burden of this complex disease. Copyright (C) 2019 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 license.Peer reviewe

The global, regional, and national burden of pancreatic cancer and its attributable risk factors in 195 countries and territories, 1990-2017: a systematic analysis for the Global Burden of Disease Study 2017

Background: Worldwide, both the incidence and death rates of pancreatic cancer are increasing. Evaluation of pancreatic cancer burden and its global, regional, and national patterns is crucial to policy making and better resource allocation for controlling pancreatic cancer risk factors, developing early detection methods, and providing faster and more effective treatments. Methods: Vital registration, vital registration sample, and cancer registry data were used to generate mortality, incidence, and disability-adjusted life-years (DALYs) estimates. We used the comparative risk assessment framework to estimate the proportion of deaths attributable to risk factors for pancreatic cancer: smoking, high fasting plasma glucose, and high body-mass index. All of the estimates were reported as counts and age-standardised rates per 100 000 person-years. 95% uncertainty intervals (UIs) were reported for all estimates. Findings: In 2017, there were 448 000 (95% UI 439 000\u2013456 000) incident cases of pancreatic cancer globally, of which 232 000 (210 000\u2013221 000; 51\ub79%) were in males. The age-standardised incidence rate was 5\ub70 (4\ub79\u20135\ub71) per 100 000 person-years in 1990 and increased to 5\ub77 (5\ub76\u20135\ub78) per 100 000 person-years in 2017. There was a 2\ub73 times increase in number of deaths for both sexes from 196 000 (193 000\u2013200 000) in 1990 to 441 000 (433 000\u2013449 000) in 2017. There was a 2\ub71 times increase in DALYs due to pancreatic cancer, increasing from 4\ub74 million (4\ub73\u20134\ub75) in 1990 to 9\ub71 million (8\ub79\u20139\ub73) in 2017. The age-standardised death rate of pancreatic cancer was highest in the high-income super-region across all years from 1990 to 2017. In 2017, the highest age-standardised death rates were observed in Greenland (17\ub74 [15\ub78\u201319\ub70] per 100 000 person-years) and Uruguay (12\ub71 [10\ub79\u201313\ub75] per 100 000 person-years). These countries also had the highest age-standardised death rates in 1990. Bangladesh (1\ub79 [1\ub75\u20132\ub73] per 100 000 person-years) had the lowest rate in 2017, and S\ue3o Tom\ue9 and Pr\uedncipe (1\ub73 [1\ub71\u20131\ub75] per 100 000 person-years) had the lowest rate in 1990. The numbers of incident cases and deaths peaked at the ages of 65\u201369 years for males and at 75\u201379 years for females. Age-standardised pancreatic cancer deaths worldwide were primarily attributable to smoking (21\ub71% [18\ub78\u201323\ub77]), high fasting plasma glucose (8\ub79% [2\ub71\u201319\ub74]), and high body-mass index (6\ub72% [2\ub75\u201311\ub74]) in 2017. Interpretation: Globally, the number of deaths, incident cases, and DALYs caused by pancreatic cancer has more than doubled from 1990 to 2017. The increase in incidence of pancreatic cancer is likely to continue as the population ages. Prevention strategies should focus on modifiable risk factors. Development of screening programmes for early detection and more effective treatment strategies for pancreatic cancer are needed. Funding: Bill &amp; Melinda Gates Foundation

Mapping subnational HIV mortality in six Latin American countries with incomplete vital registration systems

BackgroundHuman immunodeficiency virus (HIV) remains a public health priority in Latin America. While the burden of HIV is historically concentrated in urban areas and high-risk groups, subnational estimates that cover multiple countries and years are missing. This paucity is partially due to incomplete vital registration (VR) systems and statistical challenges related to estimating mortality rates in areas with low numbers of HIV deaths. In this analysis, we address this gap and provide novel estimates of the HIV mortality rate and the number of HIV deaths by age group, sex, and municipality in Brazil, Colombia, Costa Rica, Ecuador, Guatemala, and Mexico.MethodsWe performed an ecological study using VR data ranging from 2000 to 2017, dependent on individual country data availability. We modeled HIV mortality using a Bayesian spatially explicit mixed-effects regression model that incorporates prior information on VR completeness. We calibrated our results to the Global Burden of Disease Study 2017.ResultsAll countries displayed over a 40-fold difference in HIV mortality between municipalities with the highest and lowest age-standardized HIV mortality rate in the last year of study for men, and over a 20-fold difference for women. Despite decreases in national HIV mortality in all countries-apart from Ecuador-across the period of study, we found broad variation in relative changes in HIV mortality at the municipality level and increasing relative inequality over time in all countries. In all six countries included in this analysis, 50% or more HIV deaths were concentrated in fewer than 10% of municipalities in the latest year of study. In addition, national age patterns reflected shifts in mortality to older age groups-the median age group among decedents ranged from 30 to 45years of age at the municipality level in Brazil, Colombia, and Mexico in 2017.ConclusionsOur subnational estimates of HIV mortality revealed significant spatial variation and diverging local trends in HIV mortality over time and by age. This analysis provides a framework for incorporating data and uncertainty from incomplete VR systems and can help guide more geographically precise public health intervention to support HIV-related care and reduce HIV-related deaths.Peer reviewe

Mapping geographical inequalities in childhood diarrhoeal morbidity and mortality in low-income and middle-income countries, 2000–17 : analysis for the Global Burden of Disease Study 2017

Background Across low-income and middle-income countries (LMICs), one in ten deaths in children younger than 5 years is attributable to diarrhoea. The substantial between-country variation in both diarrhoea incidence and mortality is attributable to interventions that protect children, prevent infection, and treat disease. Identifying subnational regions with the highest burden and mapping associated risk factors can aid in reducing preventable childhood diarrhoea. Methods We used Bayesian model-based geostatistics and a geolocated dataset comprising 15 072 746 children younger than 5 years from 466 surveys in 94 LMICs, in combination with findings of the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2017, to estimate posterior distributions of diarrhoea prevalence, incidence, and mortality from 2000 to 2017. From these data, we estimated the burden of diarrhoea at varying subnational levels (termed units) by spatially aggregating draws, and we investigated the drivers of subnational patterns by creating aggregated risk factor estimates. Findings The greatest declines in diarrhoeal mortality were seen in south and southeast Asia and South America, where 54·0% (95% uncertainty interval [UI] 38·1–65·8), 17·4% (7·7–28·4), and 59·5% (34·2–86·9) of units, respectively, recorded decreases in deaths from diarrhoea greater than 10%. Although children in much of Africa remain at high risk of death due to diarrhoea, regions with the most deaths were outside Africa, with the highest mortality units located in Pakistan. Indonesia showed the greatest within-country geographical inequality; some regions had mortality rates nearly four times the average country rate. Reductions in mortality were correlated to improvements in water, sanitation, and hygiene (WASH) or reductions in child growth failure (CGF). Similarly, most high-risk areas had poor WASH, high CGF, or low oral rehydration therapy coverage. Interpretation By co-analysing geospatial trends in diarrhoeal burden and its key risk factors, we could assess candidate drivers of subnational death reduction. Further, by doing a counterfactual analysis of the remaining disease burden using key risk factors, we identified potential intervention strategies for vulnerable populations. In view of the demands for limited resources in LMICs, accurately quantifying the burden of diarrhoea and its drivers is important for precision public health

Mapping child growth failure across low- and middle-income countries

Childhood malnutrition is associated with high morbidity and mortality globally1. Undernourished children are more likely to experience cognitive, physical, and metabolic developmental impairments that can lead to later cardiovascular disease, reduced intellectual ability and school attainment, and reduced economic productivity in adulthood2. Child growth failure (CGF), expressed as stunting, wasting, and underweight in children under five years of age (0�59 months), is a specific subset of undernutrition characterized by insufficient height or weight against age-specific growth reference standards3�5. The prevalence of stunting, wasting, or underweight in children under five is the proportion of children with a height-for-age, weight-for-height, or weight-for-age z-score, respectively, that is more than two standard deviations below the World Health Organization�s median growth reference standards for a healthy population6. Subnational estimates of CGF report substantial heterogeneity within countries, but are available primarily at the first administrative level (for example, states or provinces)7; the uneven geographical distribution of CGF has motivated further calls for assessments that can match the local scale of many public health programmes8. Building from our previous work mapping CGF in Africa9, here we provide the first, to our knowledge, mapped high-spatial-resolution estimates of CGF indicators from 2000 to 2017 across 105 low- and middle-income countries (LMICs), where 99 of affected children live1, aggregated to policy-relevant first and second (for example, districts or counties) administrative-level units and national levels. Despite remarkable declines over the study period, many LMICs remain far from the ambitious World Health Organization Global Nutrition Targets to reduce stunting by 40 and wasting to less than 5 by 2025. Large disparities in prevalence and progress exist across and within countries; our maps identify high-prevalence areas even within nations otherwise succeeding in reducing overall CGF prevalence. By highlighting where the highest-need populations reside, these geospatial estimates can support policy-makers in planning interventions that are adapted locally and in efficiently directing resources towards reducing CGF and its health implications. © 2020, The Author(s)

Anemia prevalence in women of reproductive age in low- and middle-income countries between 2000 and 2018

Anemia is a globally widespread condition in women and is associated with reduced economic productivity and increased mortality worldwide. Here we map annual 2000–2018 geospatial estimates of anemia prevalence in women of reproductive age (15–49 years) across 82 low- and middle-income countries (LMICs), stratify anemia by severity and aggregate results to policy-relevant administrative and national levels. Additionally, we provide subnational disparity analyses to provide a comprehensive overview of anemia prevalence inequalities within these countries and predict progress toward the World Health Organization’s Global Nutrition Target (WHO GNT) to reduce anemia by half by 2030. Our results demonstrate widespread moderate improvements in overall anemia prevalence but identify only three LMICs with a high probability of achieving the WHO GNT by 2030 at a national scale, and no LMIC is expected to achieve the target in all their subnational administrative units. Our maps show where large within-country disparities occur, as well as areas likely to fall short of the WHO GNT, offering precision public health tools so that adequate resource allocation and subsequent interventions can be targeted to the most vulnerable populations.Peer reviewe