Since the early reports of neurofibrillary Tau pathology in brains of some Huntington’s disease (HD) patients, mounting evidence of multiple alterations of Tau in HD brain tissue has emerged in recent years. Such Tau alterations range from increased total levels, imbalance of isoforms generated by alternative splicing (increased 4R-/3R-Tau ratio) or by post-translational modifications such as hyperphosphorylation or truncation. Besides, the detection in HD brains of a new Tau histopathological hallmark known as Tau nuclear rods (TNRs) or Tau-positive nuclear indentations (TNIs) led to propose HD as a secondary Tauopathy. After their discovery in HD brains, TNIs have also been reported in hippocampal neurons of early Braak stage AD cases and in frontal and temporal cortical neurons of FTD-MAPT cases due to the intronic IVS10+16 mutation in the Tau gene (MAPT) which results in an increased 4R-/3R-Tau ratio similar to that observed in HD. TNIs are likely pathogenic for contributing to the disturbed nucleocytoplasmic transport observed in HD. A key question is whether correction of any of the mentioned Tau alterations might have positive therapeutic implications for HD. The beneficial effect of decreasing Tau expression in HD mouse models clearly implicates Tau in HD pathogenesis. Such beneficial effect might be exerted by diminishing the excess total levels of Tau or specifically by diminishing the excess 4R-Tau, as well as any of their downstream effects. In any case, since gene silencing drugs are under development to attenuate both Huntingtin (HTT) expression for HD and MAPT expression for FTD-MAPT, it is conceivable that the combined therapy in HD patients might be more effective than HTT silencing alone. HTT gene, while individuals with expansions of 40 or more repeats develop HD. Carriers of 36–39 CAG repeats have lower penetrance and later onset of the disease  is an autosomal dominant neurodegenerative disorder caused by a mutation in the gene that encodes for the protein Huntingtin (Htt). This mutation consists on an abnormal expansion of a CAG triplet (>35) in exon 1 of the gene that encodes for a stretch of polyglutamine (polyQ) in the N-terminal region of the protein  and in the nucleus where gene expression can get altered  that was first discovered in 1975 by Weingarten when it was co-purified along with tubulin  and/or neuropil threads  primary Tauopathies that are a major class of Frontotemporal Lobar Degeneration (FTLD) neuropathology and can present clinically with several forms of Frontotemporal Dementia (FTD)—like Frontotemporal Dementia with parkinsonism linked to chromosome 17—(FTDP-17), progressive supranuclear palsy syndrome (PSP) or corticobasal degeneration (CBD); and (b) secondary or non-primary Tauopathies like Alzheimer’s disease (AD) in which neurofibrillary Tau neuropathology occurs in addition to the amyloid-β (Aβ) plaques. The various Tauopathies affect different brain regions and cell types and they also show differences in the ratio of Tau isoforms present in the Tau filaments  was found in the cortex of HD patients and this is accompanied with the appearance of lower molecular weight (35 and 39 KD) bands  and FTLD with Tau+ inclusions (FTLD-Tau), alteration of alternative splicing of exon 10 produces an imbalance in 4R-Tau and 3R-Tau isoforms  proteins participate on constitutive splicing while also regulating alternative splicing. Some members of this family promote the inclusion of exon 10, while others suppress it. Several studies have shown that SRSF1 (ASF/SF2), SRSF2 (SC35), SRSR6 (SRp55), and SRSF9 (SRp30c) promote exon 10 inclusion, while SRSF3, SRSF4, SRSF7, and SRSF11 suppress its inclusion  could also be responsible for the 4R-Tau/3R-Tau imbalance as it has been shown to modulate exon 10 splicing and to interact with FUS, one major component of mHtt inclusions  and Tau deposits stained with antibodies that recognize 3R-Tau, 4R-Tau or pathologically phosphorylated Tau (AT-8 and pS199) has been detected in cortical and striatal sections of HD patients  or 103Q mHtt and Tau fused with non-fluorescence halves of a fluorescence reporter protein, Blum et al.  also known as Tau Nuclear Rods (TNRs) in the striatum and cortex of HD patients  Brain Bank , the Neurological Tissue Bank of the IDIBAPS Biobank , the Banco de Tejidos Fundación Cien  and the Netherlands Brain Bank . Written informed consent for brain removal after death for diagnostic and research purposes was obtained from brain donors and/or next of kin. Procedures, information and consent forms have been approved by the Bioethics Subcommittee of Centro Superior de Investigaciones Científicas .Both authors (MF-N and JL) revised the literature and wrote the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


On January 7, 2019, three patients arrived at the Community Regional Medical Center emergency department in Fresno, California, after snorting  white powder they thought was cocaine. One (patient A) was in cardiac arrest, and two (patients B and C) had opioid toxidrome  . After sAfter notification of the initial three cases, a multiagency response was implemented by FCDPH; Fresno County Sheriff-Coroner’s Office; Fresno Police Department; Fresno County Department of Behavioral Health; Community Regional Medical Center; University of San Francisco-Fresno; and the Drug Enforcement Administration. Initial actions included 1) disseminating a news release targeting the media and other emergency departments; 2) holding a multiagency press conference; 3) conducting media interviews; 4) informing law enforcement, prehospital providers, and the public about naloxone distribution and use; 5) educating persons on the proper disposal of old or new but unused medications through the Fresno County Department of Behavioral Health/California Health Collaborative drop-off containersOn January 12, 2019, a similar drug overdose incident was reported in Chico, California, in which postmortem toxicology testing for one person confirmed fentanyl (Death rates involving cocaine increased by approximately one third during 2016–2017. In 2017, nearly three fourths of cocaine deaths also involved opioids, with the arrival of synthetic opioids driving much of this increase (In the days following the multiagency press conference, FCDPH disseminated a California Health Alert Network message to approximately 700 Fresno County providers about free online medication-assisted treatment waiver training and encouraged use of the Controlled Substance Utilization Review and Evaluation System prescription drug–monitoring program for opioid users.


Variation in reproductive lifespan and female fertility have implications for health, population size and ageing. Fertility declines well before general signs of menopause and is also adversely affected by common reproductive diseases, including polycystic ovarian syndrome (PCOS) and endometriosis. Understanding the factors that regulate the timing of puberty and menopause, and the relationships with fertility are important for individuals and for policy. Substantial genetic variation exists for common traits associated with reproductive lifespan and for common diseases influencing female fertility. Genetic studies have identified mutations in genes contributing to disorders of reproduction, and in the last ten years, genome-wide association studies (GWAS) have transformed our understanding of common genetic contributions to these complex traits and diseases. These studies have made great progress towards understanding the genetic factors contributing to variation in traits and diseases influencing female fertility. The data emerging from GWAS demonstrate the utility of genetics to explain epidemiological observations, revealing shared biological pathways linking puberty timing, fertility, reproductive ageing and health outcomes. Many variants implicate DNA damage/repair genes in variation in the age at menopause with implications for follicle health and ageing. In addition to the discovery of individual genes and pathways, the increasingly powerful studies on common genetic risk factors help interpret the underlying relationships and direction of causation in the regulation of reproductive lifespan, fertility and related traits. Reproductive life span from the onset of puberty, age-specific fertility rates, and twinning frequency all contribute to fertility.12 This genetic contribution to reproductive traits and diseases can result from rare mutations in specific genes and common variation at many sites in the genome each with small effects. In the last decade, genome-wide association studies (GWAS) have transformed our understanding of genetic contributions to these complex traits and diseases.13 The results of these studies have led to discovery of novel genes and pathways influencing specific traits and diseases, new discoveries in disease epidemiology, and the discovery or repurposing of candidate therapeutics.13 Results for GWAS studies for reproductive traits , breast cancer and cardio metabolic disease.28 At the other end of the distribution, delayed puberty is associated with short stature and lower bone mineral density.29Menarche occurs with maturation of the reproductive endocrine system and denotes the onset of menstrual cycles and sexual maturity for women. It usually occurs between 9 and 14 years of age. The average age at menarche has declined over the last century in high income countries including Europe and North America.30 These include mutations in genes for kisspeptin (KISS1), GnRH (GNRH1), follicle-stimulating hormone (FSH), luteinizing hormone (LH) and their respective receptors. Some rare mutations for IHH have been implicated in less severe delayed puberty and may contribute to the population variability in AAM.11 In contrast, activating mutations in KISS1 and kisspeptin receptor (KISS1R), and paternally inherited mutations in two imprinted genes Makorin ring-finger 3 (MKRN3) and Delta-like 1 homologue (DLK1) can result in central precocious puberty (CPP).32MKRN3 is an intronless gene located on the long arm of chromosome 15 in a cluster of imprinted genes in the region associated with Prader–Willi syndrome.31MKRN3 is expressed from the paternal allele  in the arcuate nucleus and inhibits the secretion of GnRH during the prepubertal period in childhood.33 Mutations associated with CPP31 include a four-base deletion in the proximal promoter of MKRN3 identified in a girl with non-familial idiopathic CPP who exhibited pubertal development around the age of seven.34 The deletion was associated with reduced promoter activity in vitro.34DLK1 is located on chromosome 14 and encodes a transmembrane protein with multiple epidermal growth factor repeats. Like MKRN3, DLK1 is expressed from the paternal allele and paternal inheritance of a complex genomic rearrangement including the translation start site of DLK1 was associated with isolated familial CPP.32 The association of CPP with loss-of-function alleles in two paternally expressed genes supports a role for imprinting affecting the timing of puberty.32The timing of puberty varies between individuals and is a highly polygenic trait with both rare and common variants contributing to the variation in age at menarche (AAM). Rare mutations in genes that disrupt the development and function of the gonadotrophin-releasing hormone (GnRH) pathway, pituitary hormones, and their receptors can result in idiopathic hypogonadotropic hypogonadism (IHH) and the absence of puberty.30 The Lin-28 Homologue 28 (LIN28B) at 6q21 was the first of the many loci identified from five independent GWAS for AAM.35 The most recent GWAS conducted in ∼370,000 women of European ancestry identified 389 independent signals explaining ∼7.4% of the population variance in age at menarche , and genes disrupted in rare disorders of puberty GNRH1 and KISS1.36 Analyses of the data sets for genetic effects on gene expression demonstrated positive enrichment for AAM-associated variants in five central nervous system tissues, notably the pituitary gland and hypothalamus36 supporting an important role of central mechanisms in reactivation of the hypothalamic pituitary axis and onset of puberty. Expression of 205 genes in these tissues were regulated by AAM-associated variants including higher expression of LIN28B in the pituitary gland associated with later AAM.36 Formal analysis of overlap in signals for AAM and genetic effects on gene expression in blood  identified evidence for causal or pleotropic effects for 60 regions36 with the variant(s) associated with AAM driving variation of a specific genes in each region.GWAS of AAM in healthy women in European and Non-European populations have identified key genetic factors regulating puberty timing as reviewed previously.37 GWAS in 67,029 women identified 10 significant association signals for age AAM, including signals in two novel regions not reported previously in European women and one novel Japanese-specific signal in a previously reported region near PTPRD. More variants had larger effects on early, compared with late menarche timing, consistent with evidence in European studies.37In Japanese women, the mean age at menarche was 13.9 years with a strong secular trend for decreasing age at menarche to a mean age of 12.3 years for women born after 1965.38 The age at menopause (ANM) has a strong genetic component with more than 50% of variation due to genetic factors.39 A genomic analysis for age at natural menopause was conducted in ~70,000 women  with a range of effect sizes from 0.07 to 0.88 years per allele. Exome array analysis identified genome-wide significant evidence for association with two correlated low-frequency missense variants in DNA helicase B (HELB). Although interpretation of the GWAS results have limitations because specific target genes in each region are yet to be determined, the results implicate a substantial role for DNA damage repair in reproductive ageing40 including repair of double strand breaks, mismatch repair and base-excision repair. Pathway analysis also suggested enrichment for a set of genes associated with POI, including MCM8, POLG and MSH5.40Menopause is defined as the permanent cessation of menstruation resulting from the loss of follicular activity. Menopause generally occurs between ages of 40–60 years with the average age of ~51 years in western countries.NLRP11, TMEM150B and BRSK1.41 A recent GWAS study for ANM in 43,861 Japanese women identified 16 independent genome-wide significant signals , cardiovascular disease, and effects on gynaecological, gastrointestinal, musculoskeletal, and respiratory conditions.14 The relationships are complex with non-linear relationships between AAM and T2D and cardiovascular disease.42 Early menarche increases risk for both T2D and cardiovascular disease while later menarche also increases risk for cardiovascular disease, but is neutral for T2D.11 BMI also plays a role for many of the same health outcomes. Following adjustment for body composition and socio-economic status, associations with early menarche remained significant for 14 health outcomes and associations with late menarche were significant for eight health outcomes.14 Early menarche was associated with higher risks for uterine fibroids, endometriosis and earlier natural menopause.Variation in AAM and ANM have wide-ranging effects on human health. In a study in the UK Biobank, age at menarche was associated with 26 adverse health outcomes.BRCA1, MSH6 and CHEK2 that also predispose to familial cancers.39 In addition, later age at menopause results in longer exposes to high levels of oestrogen for women. Poor DNA damage response that increases cancer risk might be expected to lead to earlier menopause, opposite to the evidence from epidemiological studies. Using the available genetic data from studies for breast cancer and age at menarche, predicted age at menopause from genetic variants showed increased risks of breast cancer with later age at menopause40 and the effects were greater in oestrogen receptor positive breast cancer. Genetic variants associated with DNA damage repair genes had smaller effects than other markers.40 Taken together, the results suggest that increased exposure to oestrogen with increased reproductive lifespan is the predominant effect on risk for breast cancer.Later age at menopause is associated with increased risks for breast, ovarian and endometrial cancer. Many of the genetic markers influencing menopause are related to DNA damage repair genes including 43 Successful reproduction denoted by age at first birth (AFB) and the associated behaviour of age at first sexual intercourse (AFS) are both moderately heritable and genetically correlated.44 Estimates from the UK Biobank for SNP-based heritability for AFS and AFB were 0.242 (s.e. = 0.010) and 0.290 (s.e. = 0.015) respectively. There was a strong genetic correlation between AFS and AFB (rg = 0.86) and moderate genetic correlations between menarche and both AFS (rg = 0.22) and AFB (rg = 0.24).44 GWAS identified 34 genome-wide significant signals associated with AFS in women with replication in deCODE data for the Icelandic population, and in the Women’s Genome Health Study. The signals include association with intronic SNPs in the oestrogen receptor 1 (ESR1) gene that are also associated with AFB and with the number of children ever born. The ESR1 SNPs associated with AFB are unrelated to SNPs at this locus associated with puberty timing and breast cancer. On chromosome 3, the SNP rs2188151 associated with AFS is highly correlated with a missense variant in the semaphorin protein SEMA3F. The SNP also influences expression (is a cis-eQTL) for the RNA binding protein RMB6. The AFS decreasing allele is associated with later age at menarche, earlier AFB and greater numbers of children born.44Fertility traits in human populations are under genetic control.45 Follow-up analyses identified a number of genes in the genomic regions associated with AFB and number of children born that could be prioritised for functional studies. The critical SNPs on chromosome 1 associated with AFB and number of children ever born are correlated with likely functional nonsynonymous SNPs in two genes; the CREB-regulated transcription co-activator 2 (CRTC2) which acts downstream of FSH in ovarian granulosa cells, and CREB protein 3 like 4 (CREB3L4) which is highly expressed in reproductive tissues in both females and males.45 The lead SNP (rs2777888) for AFB on chromosome 3 is associated with altered DNA methylation or expression of several genes with a role in cell cycle progression and/or sperm function.45 Further functional experiments will be necessary to determine the specific genes and mechanisms of action of the large number of genetic variants influencing these important traits.A large study of genetic effects on reproductive behaviour in 62 cohorts of European ancestry  identified 10 independent genomic loci associated with AFB in women, men or both.46 POI is generally defined as the onset of menopause in women under the age of 40 years. Genetic mutations have been reported in a number of candidate genes, although many reported results have not been replicated.47 There is good evidence for deleterious effects of mutations in Bone Morphogenetic Protein 15 (BMP15), Progesterone receptor membrane component 1 (PGRMC1) and the pre-mutation in the Fragile-X mental retardation 1 (FMR1) locus on the X chromosome.47 Mutations in other genes, present in low frequencies in some populations, are likely to influence POI including GDF9, Folliculogenesis specific bHLH transcription factor (FIGLA), and Newborn ovary homeobox gene (NOBOX).47 The 5-prime untranslated region of the FMR1 gene contains a CGC repeat that varies in length and expansion of the repeat to >200 copies causes Fragile-X syndrome. Repeat expansion to the pre-mutation range is associated with premature menopause, but there is no influence of shorter repeat lengths in the normal range or the longer repeats that cause Fragile-X syndrome.48Ovarian reserve is one of the crucial elements of female fertility Fig.  and seve47 Although GWAS have identified multiple loci associated with POI in Chinese, Korean, and Dutch women,47 the small samples studied (<1000) have low statistical power and none identified signals exceeding 10−6. Pathway analysis of suggestive GWAS loci for ANM do show enrichment for known POI genes.39 There are ~80 gene disorders influencing extremes of reproductive function. The genes are involved in diverse biological processes, including control of the cell cycle, DNA damage response and repair, hormone signalling and gonadal development. Genetic variants in or near many of these genes have variable effects from very early menopause to alterations of just a few weeks.39 Future studies involving larger data sets and meta-analysis of combined GWAS will be necessary to identify risk loci associated with POF.A total of six GWAS have been conducted to identify the risk loci in POI in different populations as reviewed in detail elsewhere.49 DZ twins arise from the ovulation of two follicles after a complex process of follicle growth, selection and ovulation. The frequency of DZ twinning ranges from 6–15 per 1000 live births and varies with maternal age and ethnicity.50 DZ twinning is influenced by genetic factors and the DZ twinning rate for sisters of women with spontaneous DZ twins is ~2.5 times higher than the twinning rate in the general population.50 Direct evidence for genetic variants influencing rates of DZ twinning were first identified in animal studies. Domestic sheep generally have 1–2 offspring at each pregnancy. Linkage and positional cloning identified mutations in autosomal and X-linked genes that increased ovulation and twinning rates in heterozygous carriers.53 The mutations responsible were identified in the genes for Bone Morphogenetic Protein Receptor Type 1B (BMPR1B) and BMP15, respectively; a receptor and ligand in the ovarian signalling pathway for BMP15.53 Multiple mutations have now been reported in both BMP15 and the closely related signalling molecule growth differentiation factor 9 (GDF9).54 Physiological and genotype-phenotype studies of strains carrying the different mutations demonstrate the importance of BMP signalling, and the balance between BMP15 and GDF9 in follicle survival, maturation, and control of ovulation.54The spontaneous dizygotic (DZ) twinning rate is associated with fertility and comparisons with the frequency of monozygotic twins provide a useful index of fertility in a population.BMP15 and GDF9 have alternative phenotypes depending on whether they are heterozygous or homozygous (carried as one or two copies respectively). Increased ovulation rates occur in heterozygous carriers of the mutations as described above. In contrast, homozygous carriers with two copies of the loss-of-function mutations in BMP15 and GDF9 in sheep have streak ovaries and are completely infertile.54 These genes are candidates for infertility in women and studies in women with primary ovarian insufficiency (POI), also known as premature ovarian failure (POF), have identified mutations in both genes associated with POI.55 Confirmation through segregation studies is difficult in families with infertility disorders, but molecular and functional studies for several of these variants suggest loss-of-function effects consistent with the view that two functional copies of BMP15 are required for an adequate ovarian reserve in women.55 Low-frequency variants in GDF9 are associated with increased risk for DZ twinning in women heterozygous for these variants.56 Similar studies found no evidence that rare and low-frequency variants in BMP15 influence DZ twinning.57 Mutation screening of BMPR1B in mothers of DZ twins identified a coding variant (p.Gln294Glu) altering the same amino acid as a sheep BMPR1B functional variant.58 This mutation was not seen in other families and is of unknown significance.Some mutations in 56 In a search for common genetic variants influencing DZ twinning, a GWAS in 1980 mothers of spontaneous DZ twins and 12,953 controls identified significant association with DZ twinning for SNPs close to Follicle-Stimulating Hormone Beta Subunit (FSHB) and SMAD Family Member 3 (SMAD3).59 The risk alleles for the SNPs close to FSHB and SMAD3 increased the frequency of twin births in the Icelandic population by 18 and 9%, respectively. The lead SNP associated with DZ twinning on chromosome 15 maps to the first intron of SMAD3, strongly expressed in the human ovary, where it promotes granulosa cell proliferation and steroidogenesis.59 The region of chromosome 15q22.33 also includes SMAD Family Member 6 (SMAD6). A major gene increasing ovulation rate and twinning in cattle maps to the equivalent genomic region on bovine chromosome 10 (the location of both SMAD3 and SMAD6).60 Recent analysis of gene expression in granulosa cells from carriers and non-carriers of the cattle gene demonstrated a six-fold increase in expression of SMAD6 in gene carriers.61 SMAD6 is an inhibitor of BMP/SMAD signalling and over-expression of SMAD6 is consistent with loss-of-function mutations in BMP signalling increasing ovulation rate in sheep. Further research is required to determine whether variants associated with DZ twinning on chromosome 15 act through effects on SMAD3, SMAD6 or some other mechanism.Rare and low-frequency variants account for only a small proportion of variation in DZ twinning.63 PCOS is the most common cause of infertility64 and also increases the risk for type 2 diabetes, gestational diabetes, venous thromboembolism, cerebrovascular and cardiovascular disease and endometrial adenocarcinoma.65 The aetiology of PCOS remains unclear with diagnostic criteria proposed for PCOS66 including the National Institutes of Health (NIH), Rotterdam and Androgen Excess Society (AES) criteria. Familial aggregation and twin studies suggest genetic factors play a strong role in pathogenesis of PCOS with heritability estimates of 70%.66Polycystic ovary syndrome (PCOS) is a complex, hormonal and metabolic disorder affecting 5–20% of women of reproductive age globally and characterised by hyperandrogenism, ovulatory dysfunction, polycystic ovarian morphology and gonadotropic abnormalities.67 Additional studies69 identify 16 independent signals in 15 genomic regions associated with PCOS including signals near important reproductive hormone genes FSHR, LHCGR and FSHB.70 The signals also include variants in or near three epidermal growth factor genes and genes associated with diabetes.70 As with similar studies in other reproductive diseases, the total heritability explained by GWAS identified PCOS risk SNPs is relatively low (<10%).65The first GWAS conducted in Chinese patients in 2011  has direct clinical applications for planning lifestyle modification as a prevention strategy and inclusion of metformin in treatment plans for PCOS.68 Discovery of additional genetic factors and further characterisation of the signals identified will provide greater insight into the pathogenesis of the complex phenotypes in PCOS.Mendelian randomisation is an analytical method using genetic variation to investigate the likely causal relationship between an exposure trait (or risk factor) and a health outcome. Genetic variants are inherited independently and fixed at birth and subject to less confounding than other measured risk factors. Increasing evidence of association between genetic variants and many common traits means Mendelian randomisation models can use genetic variants associated with a risk factor to infer relationships with a health outcome. The models assume the genetic variants used have strong evidence for association with the risk factor and do not influence the outcome through other unrelated biological pathways. In PCOS studies, Mendelian randomisation demonstrated causal roles for higher BMI, greater insulin resistance and reduced sex hormone binding globulin concentrations in serum.7 The disease affects 7–10% of women and is associated with pain and infertility. Early twin studies identified evidence for genetic effects on the liability for hysterectomy,71 one of the most commonly performed surgical procedures for women. Major indications for hysterectomies in reproductive age women are endometriosis (30%) and uterine leiomyomas (>50%).73 Subsequent studies in twins on genetic influences on the liability for endometriosis estimated the heritability at ~50%.75Endometriosis is a complex disease characterised by ectopic lesions of tissue resembling endometrium in the peritoneal cavity.78 The most recent meta-analysis  is the predominant receptor for oestrogen action in the endometrium.80 Genomic signals associated with endometriosis include regions flanking the gene for ESR1, signals upstream of follicle-stimulating hormone beta subunit (FSHB) known to increase FSH concentrations,79 and near the oestrogen-regulated and early response gene (GREB1) first identified in breast cancer cell lines and tumours.81 Other regions include candidate genes with roles in cell migration, adhesion and proliferation including Cell Division Cycle 42 (CDC42), Cyclin-Dependent Kinase Inhibitor 2B Antisense (CDKN2B-AS1) and Kinase insert domain receptor (KDR).84Genomic regions and genes associated with endometriosis risk are reviewed in detail elsewhere.sis Fig.  identifi85 On chromosome 1, studies on genetic regulation of gene expression in blood86 and endometrium86 show the critical SNPs in this region influence expression of both the long non-coding RNA LINC00339 and CDC42. There are chromatin interactions between risk SNPs and gene promoters for both LINC00339 and CDC42.86 Luciferase reporter assays support the effect of genetic differences on the interaction between the regulatory element and the promoter of CDC42.86 Formal analysis of the overlap of signals for endometriosis risk and genetic effects on gene expression provides strong evidence that key SNPs associated with endometriosis on chromosome 1 and chromosome 12 regulate LINC00339 and vezatin VEZT expression, respectively.87Mechanisms leading to formation of lesions are poorly understand, but one source for cells initiating these lesions is thought to be cells shed from the endometrium and deposited in the pelvic cavity through retrograde menstruation.TACR3 and age at menopause), rare, low-frequency and common variants all influence the same trait.11 In other examples , rare variants cause hypogonadotropic hypogonadism while common variants influence age at menopause.11 These differences, and evidence for the different effects on ovulation rate and streak ovaries for heterozygous and homozygous mutations in BMP15 and GDF9 in sheep54 show disruption of gene function or altered regulation of the same genes can have different effects on reproductive traits and diseases.Genetic studies discussed above demonstrate the complex variation contributing to the timing of puberty, menopause, ovarian function and twinning. Genetic contributions include the effects of multiple common variants with small effects on reproductive traits and disease, and rare variants with large effects contributing to failures in development, precocious puberty, delayed puberty, premature ovarian failure and increased twinning. In some cases  and genes associated with POI.40 Further research will help to understand the relationships between ovarian development, disorders of puberty, DZ twinning, POI, and age at natural menopause.Potential overlap for other traits includes genes influencing both DZ twinning and POI and endometriosis and ovarian cancer. There is a small, but significant increase in mothers of DZ twins reaching menopause before the age of 40 compared with mothers of MZ twins.92 The association signals are located in WNT4.92 Subsequent follow-up functional studies show correlated association signals for endometriosis in this region regulate LINC00339 and CDC42 not WNT4,83 and one or both target genes may also play a role in ovarian cancer risk. There is genetic association near the oestrogen receptor 1 (ESR1) locus with four independent signals for endometriosis and five independent signals for breast cancer93  overlaps completely with an association signal for ovarian cancer.94 The Polygenic risk score (PRS) combines association results from genome-wide genotyping into a single estimate of the genetic risk for a disease or trait for each individual. The PRS score is calculated from the number of risk alleles carried, weighted by the effect size estimated from the discovery sample.94 It is often standardised to a mean of zero with a standard deviation of 1 for ease of interpretation.94 The risk scores are not very informative for individual prediction, but explain sufficient variation to determine individuals at highest or lowest risk in populations, and to understand genetic contributions to related traits.13 Where samples are fully independent, calculating genetic contributions to related traits provides a powerful design reducing issues of shared environment and ascertainment in epidemiological studies. Overlap between endometriosis and ovarian cancer extends beyond the observed overlap at the chromosome 1p36 locus discussed above. Observations from epidemiological studies can be confounded due to diagnosis of endometriosis at laparoscopy close to a diagnosis of ovarian cancer. PRS analysis with genotype data from independent samples for endometriosis and ovarian cancer showed shared genetic risk between endometriosis and most histotypes of ovarian cancer95 suggesting some common molecular pathways for the two diseases including the overlap at chromosome 1p36. Prediction of individual risk is improving for some diseases with better estimates of the genetic risk factors. While not sufficiently predictive in isolation, risk scores may be used in combination with other clinical data for patient stratification. The application of risk prediction to inform breast cancer screening is being evaluated in the population based Women Informed to Screen Depending On Measures of risk (WISDOM) study.96In addition to discovery and overlap in effects for individual genes, GWAS data provides powerful approaches to understand shared genetic risk between traits and diseases. Genome-wide SNP genotype data from a discovery sample can be used to estimate the genetic variation due to common SNPs or SNP heritability, and to calculate a polygenic risk score for individuals in an independent sample.97 and number of children ever born.97 Results show a significant negative genetic correlation between number of children ever born and the age at first birth suggesting genes contributing to later first birth are associated with fewer children.97 These relationships are complex because environmental effects and demographic changes also play a role with women choosing to delay childbearing in many countries. Relationships between age at first birth and lifetime number of children are also associated with genetic effects on twinning. Mothers of twins tend to have an earlier age at first birth and raise more children to adulthood in favourable environments.99 Polygenic risk score analysis from the GWAS results for DZ twinning suggest genes contributing to DZ twinning may partly explain genetic effects on fertility. The polygenic risk score for DZ twinning was significantly associated with DZ twinning in an independent sample from Iceland.59 In this sample, the risk score for DZ twinning was associated with a higher likelihood of having children, earlier age at first birth, and greater number of children.59As noted earlier, there are significant additive genetic effects for age at first birth100 This epidemiological evidence is supported by the Mendelian randomisation analyses demonstrating the causal relationship between delayed natural menopause and increased breast cancer risk.40 Understanding these relationships is further complicated by effects of variation in body mass index (BMI). There is a strong inverse genetic correlation between AAM and BMI.36 Thirty-nine signals overlap between AAM and BMI, but most AAM signals make some contribution to adult BMI.36 Using information from different genetic studies allowed estimates of the direct effects of AAM on sex-steroid-sensitive cancers after adjusting for genetically predicted BMI. In these models, later AAM was associated with reduced risks for breast, ovarian and endometrial cancers.36 Analysis of cancer subtypes suggests effects were most strongly associated with oestrogen receptor positive breast cancer and serous ovarian cancer. The effects of earlier puberty timing on higher risks of the sex- steroid sensitive cancers might be related to longer duration of exposure to sex steroids and/or increased activity of the hypothalamic-pituitary axis associated with earlier puberty and reproductive traits including dizygotic twinning.Genetic studies can also inform complex relationships between reproductive traits and related health outcomes. A large-scale meta-analysis supported an association of early menarche and late menopause with increased risk of breast cancer and demonstrated that excess risk associated with advancing menarche by one year was higher than excess risk associated with lengthening menopause by one year.Recent studies have made great progress towards understanding the genetic factors contributing to variation in traits and diseases influencing female fertility. These genetic studies are providing important insights into the complex biology contributing to female fertility. The data emerging from GWAS demonstrate the utility of genetics to explain epidemiological observations, revealing shared biological pathways linking puberty timing, fertility, reproductive ageing and health outcomes. For example, many variants influencing menopause suggest genes and pathways with known roles in DNA damage/repair with implications for follicle health and ageing, although we do not yet have a full picture of the mechanisms responsible.The effect sizes for common variants are small and, as emphasised in this and other reviews, large studies are essential to identify a substantial fraction of common variants underlying variation for these traits and diseases. In addition, GWAS identify genomic regions, but not the specific genes and pathways regulating variation in traits and diseases. To date, we lack definitive evidence for the causal variants and target genes for most of these results. Despite these gaps, examination of potential candidate genes in the multiple regions identified through GWAS provide tantalising evidence for common pathways affecting reproductive lifespan and fertility.101 This in turn will provide much better understanding of the specific pathways regulating individual components of reproductive lifespan and fertility, and the complex interactions between pathways and health outcomes. The knowledge gained will suggest possible interventions and ways to better predict and minimise health impacts of these important life events.The challenge now is to provide functional evidence for the specific genes and pathways regulated by the genetic variation influencing fertility. Multiple lines of evidence from both functional and genomic studies will be required to identify these gene targets and the mechanisms responsible. This is becoming more feasible with advances in genomics, large-scale publically available data, and genome editing.Genetic risk scores from common variants are not able to predict disease risk for individuals. As the estimates improve from more powerful studies, they may be able to predict women at higher or lower risk for specific diseases and this may be useful in some diagnostic settings. Genetic variants associated with natural menopausal timing also influence the menopausal timing in women undergoing radiation or chemotherapy suggesting that in the future, genetic risk prediction models could be useful for counselling related to family planning, lifestyle choices or use of modern techniques for fertility preservation such as oocyte cryopreservation.Epidemiological evidence for comorbidity between traits can be difficult to interpret because of the issues of sample ascertainment. Genetic risk scores add a valuable method to gain insight into the complex relationships in the overlap between traits and diseases. These analyses will also improve as the risk scores can be derived from increasingly powerful studies to help interpret the underlying relationships and direction of causation in the regulation of reproductive lifespan, fertility and related traits.


The USDA Forest Service and University of California researchers have developed a simple and inexpensive passive sampler for monitoring air concentrations of HNO3. Nitric acid is selectively absorbed on 47-mm Nylasorb nylon filters with no interference from particulate NO3-. Concentrations determined with the passive samplers closely corresponded with those measured with the co-located honeycomb annular denuder systems. The PVC protective caps of standardized dimensions protect nylon filters from rain and wind and allow for reliable measurements of ambient HNO3 concentrations. The described samplers have been successfully used in Sequoia National Park, the San Bernardino Mountains, and on Mammoth Mountain in California.Nitric acid (HNO


The authors note that the citation of Wright (1937) and Nei (1968) is incorrect in the “Models of mutation-selection balance” section in Results. Wright (1937) not only showed the equilibrium frequency of a deleterious allele for a population of infinite size (Eq 1) but also derived the full equilibrium frequency distribution of a deleterious allele under a finite effective population size. He also derived the mean (Eq 2) allele frequency of a fully recessive deleterious allele and gave approximations for large and small effective population sizes. Nei (1968) further derived the approximate variance (Eq 3) in allele frequency of a fully recessive mutation by using Wright’s general formula. The authors confirm that this error does not affect the conclusions of the article.


Scientific Reports 10.1038/s41598-019-54295-8, published online 29 November 2019Correction to: The original version of this Article contained a typographical error in the Abstract.“According to the Dual Dodel of Social Hierarchy, one pathway for attaining social status is through dominance (coercion and intimidation).”now reads:“According to the Dual Model of Social Hierarchy, one pathway for attaining social status is through dominance (coercion and intimidation).”This error has now been corrected in the PDF and HTML versions of the Article.


With the current global downturn, the organizations need to develop new strategies and innovative approaches to ensure that every aspect of sustainability is achieved. For this purpose, the organizations need an indicator that measures the fitness if an organization. The purpose of this project is to analyze the ‘Fitness’ of an organization using the dataset related to leanness, agility and sustainability in ANFIS (Adaptive Neuro-Fuzzy Inference System) in order to determine whether the company is fit enough to sustain in global markets or not. The project does so by integrating both neural networks and fuzzy logic principles with lean, agility and sustainability principles. FIT manufacturing is the integration of Lean, Agile and sustainability manufacturing in one system as a whole which would help in attaining maximum output and sustain effectively in global markets. FIT Manufacturing adopts an integrated approach towards the use of Lean, Agility and Sustainability to achieve a level of fitness that is unique to each company. The database in the paper contains lean, agile and sustainable indices reviewed by experts. FIT does not prescribe that every aspect of Lean, Agile and Sustainability methodologies must be applied to every company, but a selective mix of components will provide the optimum conditions for a company to prosper. Specifications TableValue of the data•Markets all over the globe are getting more and more competitive through the changes driven by emerging technologies which creates the need for an organization to be adaptive and compatible in order to dominate the market.•FIT manufacturing is a perfect amalgamation of core principles of lean, agile and sustainable manufacturing.•The data in this article helps to determine lean, agile and sustainability indices which provides an insight as to how to fit the organization is by providing a framework for the calculation of fitness index.•Using this data, the efficiency and responsiveness of an organization towards the constantly changing markets can be analyzed.1This dataset consists of 5 fitness enablers, 69 fitness criteria and 205 fitness attributes. The data consists of all major dimensions or enablers of fitness such as management responsibility, manufacturing management, manufacturing technology, workforce and manufacturing strategy. The data on agile, lean and sustainability criteria, was collected from automobile components manufacturing company located in Tamil Nadu, India arranged in a matrix form , Table 52The methodologies that allowed the data presented are described in 2.1The data presented in the paper was collected from automobile components manufacturing company located in Tamil Nadu. India. The attributes pertaining to leanness, agility and sustainability criteria were obtained and rated on a scale from 1 to 10 by industry experts.2.2I.If (agility is not_agile) and (leanness is not_lean) and (sustainability is not_sustainable) then (fitness is not_fit) (1)II.If (agility is average) and (leanness is average) and (sustainability is average) then (fitness is avergge) (1)III.If (agility is agile) and (leanness is lean) and (sustainability is sustainable) then (fitness is fit) (1)IV.If (agility is agile) and (leanness is average) and (sustainability is sustainable) then (fitness is fit) (1)V.If (agility is agile) and (leanness is average) and (sustainability is average) then (fitness is avergge) (1)VI.If (agility is average) and (leanness is lean) and (sustainability is average) then (fitness is avergge) (1)VII.If (agility is average) and (leanness is lean) and (sustainability is sustainable) then (fitness is fit) (1)VIII.If (agility is average) and (leanness is average) and (sustainability is sustainable) then (fitness is fit) (1)IX.If (agility is average) and (leanness is average) and (sustainability is not_sustainable) then (fitness is avergge) (1)X.If (agility is average) and (leanness is not_lean) and (sustainability is not_sustainable) then (fitness is not_fit) (1)XI.If (agility is not_agile) and (leanness is average) and (sustainability is not_sustainable) then (fitness is not_fit) (1)XII.If (agility is not_agile) and (leanness is average) and (sustainability is average) then (fitness is avergge) (1)XIII.If (agility is average) and (leanness is average) and (sustainability is not_sustainable) then (fitness is avergge) (1)XIV.If (agility is not_agile) and (leanness is average) and (sustainability is sustainable) then (fitness is avergge) (1)XV.If (agility is agile) and (leanness is average) and (sustainability is not_sustainable) then (fitness is avergge) (1)XVI.If (agility is agile) and (leanness is not_lean) and (sustainability is sustainable) then (fitness is avergge) (1)XVII.If (agility is agile) and (leanness is not_lean) and (sustainability is not_sustainable) then (fitness is not_fit) (1)XVIII.If (agility is not_agile) and (leanness is not_lean) and (sustainability is sustainable) then (fitness is not_fit) (1)XIX.If (agility is not_agile) and (leanness is lean) and (sustainability is not_sustainable) then (fitness is not_fit) (1)XX.If (agility is not_agile) and (leanness is lean) and (sustainability is not_sustainable) then (fitness is not_fit) (1)2.3MATLAB 2016 was used for creation of Adaptive Neuro-Fuzzy Inference System. The ANFIS model was created using fuzzy logic designer with the corresponding matrices as input , Fig. 5.2.4See 3FIT manufacturing is the integration of Lean, Agile and sustainability manufacturing in one system which would help in attaining maximum output and prevent the damage caused to the environment. This paper deals with analyzing the ‘fitness’ of a company using ANFIS. The integration of the Fit principles can be employed to reduce the unnecessary cost and increase efficiency of production with less wastage in a market having mercurial demand patterns. The fitness index can be used as a benchmark to evaluate overall efficiency of an organisation.


The Art to Life (ATL) program aims to improve the quality of life for persons living with dementia (PWD) through art therapy, intergenerational contact with college students, and life story preservation within an adult day service. This poster will present the results of an ongoing program evaluation to determine the effects of the intervention on PWDs’ engagement in (1) communication with others and (2) art/creative activity. A two-member analysis team independently coded ethnographic field notes utilizing operational definitions of PWDs’ observed behavior during momentary time sampling, and recording events of communication and art engagement using the modified ENGAGE measure . Results across sessions (N=97) reveal communication engagement  significantly exceeded art engagement , t (96)=20.85, p=0.001). These results suggest that engagement in reminiscence via intergenerational contact is a fundamental feature in comparison to art and creative activity within the ATL program. Exploratory qualitative content analysis of ethnographic field notes by a three-member coding team identified two key themes within the communication engagements. These emergent themes included validation of personhood and reminiscence of family ties. More studies are needed to determine if the scope and reach of intergenerational interventions may be increased through the nationwide translation of the ATL program.


An outbreak of feline panleukopaenia virus (FPLV) infection was diagnosed by pathology, electron microscopy and polymerase chain reaction (PCR) in vaccinated captive-bred subadult cheetahs in South Africa. Subsequent to this disease outbreak, 12 cases of FPLV diagnosed on histology were confirmed by PCR in captive African black-footed cat, caracal, cheetah, lion, ocelot and serval. Phylogenetic analyses of the viral capsid protein gene on PCR-positive samples, vaccine and National Center for Biotechnology Information (NCBI) reference strains identified a previously unknown strain of FPLV, present since at least 2006, that differs from both the inactivated and the modified live vaccine strains. A previously described South African strain from domestic cats and cheetahs was identified in a serval. Surveys of FPLV strains in South African felids are needed to determine the geographical and host species distribution of this virus. Since non-domestic species may be reservoirs of parvoviruses, and since these viruses readily change host specificity, the risks of FPLV transmission between captive-bred and free-ranging carnivores and domestic cats and dogs warrant further research. Felis domesticus). FPLVs are classified under the family Parvoviridae and are related to mink enteritis virus (MEV) and canine parvovirus (CPV)  infection, the first viral disease identified in cats, is one of the most common viral diseases of domestic cats (Acinonyx jubatus)   , ocelot , puma (Puma concolor), Siberian tiger , and spotted cat (Leopardus tigrinus) , arctic fox (Vulpes lagopus) and raccoon (Procyon lotor), and is genetically very similar but distinct from CPV-like viruses from dogs (Canis familiaris) and raccoons , seven cheetahs, three lions, one ocelot and two servals . Clinical signs and pathological findings were not available from many of these cases. Before the 2012 cheetah outbreak molecular diagnostic confirmation of suspected FPLV cases was not readily available in South Africa. To investigate prior suspected or confirmed cases of FPLV molecular diagnostic testing was performed on archived tissue samples from felids in the NZG Biobank (Cases A–G), including both suspected and histologically typical FPLV cases. Cases were from Gauteng and North West provinces and included one African black-footed cat (Felis nigripes), a domestic cat, three cheetahs and two lions. Samples from a puma, a leopard (Panthera pardus) and an ocelot in which clinical and histological evidence of FPLV was absent (Cases i–iii) were included as negative control cases. DNA was also extracted from current Fellocell® and Fel-O-Vax®  vaccines. All the samples are archived at the NZG Biobank.Subsequent to the outbreak in cheetahs, animal carcasses, a rectal swab, formalin-fixed tissues or formalin-fixed paraffin-embedded tissues (FFPE) from various single or multiple cases in captive felids were submitted to the NZG for pathological examination and/or molecular diagnostic testing for FPLV infection by the owners, veterinarians and pathologists throughout South Africa (Cases 6–20). Submitted cases of suspected or confirmed FPLV based on clinical signs and typical macroscopic and histological findings, or diseases presenting with similar signs, included two caracals (TM Tissue MiniPrep kit (Zymo Research). DNA from FFPE samples were extracted using the MasterPure™ Complete DNA & RNA Purification Kit (Epicentre) following the manufacturers specifications. Molecular analysis of the parvovirus genome from all the isolates was conducted by amplification of a ~1300 base pair (bp) region of the viral capsid protein (VP2) gene  treatment, cycle sequenced using the BigDye V3.1 (Applied Biosystems) chemistry and run on a 3130 genetic analyser (Applied Biosystems). The resulting sequences were analysed and aligned using the ClustalX  parvovirus reference strains. Results from NJ (10 000 bootstrap replicates) and BA (1 million generations) are presented in Clinical signs were described in detail only in the initial cheetah outbreak (Cases 1–5) and included pyrexia, haemorrhagic diathesis , hypersaOn necropsy examination, the cheetah cubs showed similar lesions including variable dehydration, mild acute diffuse tissue congestion, and moderate acute intestinal serosal congestion and haemorrhage . VariablEscherichia coli was obtained from the intestine from Case 1 and normal intestinal bacterial flora from Case 3. On negative contrast electron microscopy intestinal contents contained small numbers of parvovirus particles (Case 1).Histologically, extensive necrosis of the intestinal epithelium was accompanied by villous collapse, marked hyperplasia of scanty remaining epithelium, and a thick coat of fibrin, necrotic cellular debris and myriad bacteria on the mucosal surface . In addiThe pathological lesions were consistent with FPLV infection with secondary bacterial enteritis and septicaemia. Additional pathological findings included moderate diffuse acute interstitial pneumonia with moderate multifocal acute pulmonary oedema ; multifocal acute moderate necrohemorrhagic bacterial gastritis as well as lingual and gastric candidiasis, mild serosanguinous hydropericardium and hydrothorax and acute moderate segmental renal tubular necrosis (Case 2); oesophageal and gastric candidiasis and acute mild multifocal pancreatic necrosis (Case 3); and mild acute multifocal necrotising aspiration pneumonia (Case 5). All the cubs were in good body condition at the time of death and showed mild to moderate diffuse adrenocortical hyperplasia.® vaccine. Clade 2 is represented in the current dataset by one sample (Case 10), which clustered with previously described South African strains of FPLV isolated from a domestic cat (1999) and two cheetahs (2000) as well as an FPLV strain isolated from a wildcat and a mink in the USA (1995). This strain is 99.99% similar to the Fellocell® vaccine . All the samples from this study clustered separately from CPV (98% similarity) and clustered in two clades. Most of the positive cases (Clade 1) clustered closely (99.94% – 100%) with reference samples  from carnivores in Japan and the United States of America (USA), and were similar to the Fell-O-Vax vaccine .Clinical features, pathology, electron microscopy and molecular diagnostic tests confirmed the presence of a previously unreported strain of FPLV in the cheetah cubs in the initial outbreak (Cases 1–4). This strain was also detected in archived samples from a lion that died in 2006 (Case A) and an African black-footed cat that died in 2007 (Case B), as well as in samples submitted from other captive felids that died throughout the country after November 2012, including cheetahs , a caracal (Cases 15), a lion (Case 14), an ocelot (Case 12) and a serval (Case 6).et al. The testing of archived samples confirms that the strain from Clade 1 was present in felids in at least North West province before the initial cheetah outbreak and Clade 1 virus subsequently caused disease in unvaccinated felids in Gauteng and North West. Clade 1 strain apparently co-exists with the previously described strain , F10 products  or swimming pool chlorine (2.67 g/L water). Biosecurity measures included staff movement from clean to infected enclosures for feeding and inspections; new and frequently changed cleaning cloths as well as overalls and boots for all staff; hand gels and body sprays of F10 products used when moving between enclosures.et al. Positive cases were detected from fresh and frozen tissue, a faecal swab and FFPE intestinal samples from 16 animals clinically and pathologically diagnosed as suffering from FPLV or suspected FPLV . Three cAlthough infections with viruses of the feline parvovirus subgroup have been reported before in several wild carnivore species, including African felids, this study reveals the first in-depth insight into strain diversity within captive felids in South Africa and the first documented cases of FPLV infection in the African black-footed cat and caracal. Further surveys are needed to determine the geographical and host species distribution as well as the genetic variability of FPLV in these species. Vaccine failure, inadequate vaccination and stress may be predisposing factors in the susceptibility to disease. Retention of the current vaccination programme as well as good hygiene practices at breeding and rearing facilities is recommended for the prevention and control of the disease.


Containerized plant production represents an extremely intensive agricultural practice; 40,000 to 300,000 containers may occupy one acre of surface area to which a large amount of chemical fertilizer is applied. Currently, recommended fertilizer application rates for the production of containerized nursery ornamental plants are in excess of plant requirements, and up to 50% of the applied fertilizers may run off or be leached from containers. Among the nutrients leached or allowed to runoff, nitrogen (N) is the most abundant and is of major concern as the source of ground and surface water pollution. In this report, current N fertilizer application rates for different container-grown nursery ornamental plants, the amount of nitrate leaching or runoff from containers, and the potential for nitrate contamination of ground and surface water are discussed. In contrast, our best N management practices include: (1) applying fertilizers based on plant species need; (2) improving potting medium's nutrient holding capacity using obscure mineral additives; (3) using controlled-release fertilizers; and (4) implementing zero runoff irrigation or fertigation delivery systems that significantly reduce nitrate leaching or runoff in containerized plant production and encourage dramatic changes in N management.


PRNP reported from several countries. The objective of this study was to investigate PRNP polymorphisms of native Ethiopian goat breeds and compare the results with other goat breeds.Polymorphisms of the prion protein gene may influence scrapie susceptibility in small ruminants through modified protein conformation. At least 47 amino acid substitutions and 19 silent polymorphisms have been described in goat PRNP in 229 goats belonging to three of the main Ethiopian native goat breeds showed a remarkably high frequency (> 34.6%) of p.(Asn146Ser) in these breeds, a variant involved in scrapie resistance in Cyprus. In addition, two novel amino-acid substitutions p.(Gly127Ala) and p.(Thr193Ile), with frequencies ranging from 1.5 to 7.3% were detected. Both amino acids are well conserved in prion proteins (PrP) of most species and these changes have never been reported before in goats worldwide. Residue 127 is within the N–terminal domain of PrP and is probably involved in the recruitment of neural cell adhesion molecules (NCAM). Residue 193 is within the highly conserved string of 4 threonines that plays a role in determining the efficiency of prion protein conversion towards its pathological form.The analysis of the prion protein gene PRNP of Ethiopian goats. This finding increases the interest in exploring PRNP polymorphisms of native goat breeds in areas where cross breeding with foreign goats has rarely occurred.Two novel coding polymorphisms and a high frequency of a scrapie protective variant indicate a high level of genetic diversity in The online version of this article (10.1186/s12917-019-1870-4) contains supplementary material, which is available to authorized users. Camelus dromedarius) in Algeria enlarging the spectrum of animal species susceptible to prion disease [Scrapie is a fatal, neurodegenerative disease that affects small ruminants world-wide. It belongs to the group of prion diseases or transmissible spongiform encephalopathies (TSEs), such as bovine spongiform encephalopathy (BSE) in cattle, chronic wasting disease (CWD) in cervids and Creutzfeldt–Jakob disease (CJD) in humans. Recently, a new prion disease, Camel Prion Disease (CPD) of unknown origin was detected in dromedary camels ( disease .PRNP) sequences may influence scrapie susceptibility in sheep and goats through modified protein conformation [The conversion of normal prion protein (PrP) into pathogenic PrP conformers is central to prion diseases. Polymorphisms of the prion protein gene . The variant p.(Gly127Ala) showed a frequency of 4.1% in Western Highland goats and lower frequencies in Central Highland and Long-eared Somali goats respectively. The variant p.(Thr193Ile) showed higher frequencies in Western and Central Highland breeds and a lower frequency in the Long-eared Somali goat  are reported for Ethiopian goat breeds. The novel variant p. Gly127Ala) is located within the N terminal domain of PrP involved in the interaction with the neuronal cell adhesion molecule (NCAM) . A varia7Ala is lf Cyprus  but absef Cyprus . Both isf Cyprus .The presence of p.(Asn146Ser) in Halep (Damascus), Ethiopian and Tanzanian native goats suggests that the allele was naturally selected in the Middle East first, and spread through the central Sahara and Ethiopia later , 36 and PRNP of Ethiopian goats compared to other breeds. However, a low genetic variation among indigenous Ethiopian goats has been detected through genetic studies on microsatellites probably due to the ancient and widespread practice of nomadism [PRNP substitution frequencies among the breeds in this study. In any case extensive cross breeding with goats coming from other countries has never been performed in Ethiopia, except for some pilot studies with Saanen goats to increase milk production [The frequency of the protective variant p.Asn146Ser) in three of the main Ethiopian goat breeds and the presence of two novel amino acid substitutions in highly conserved regions of prion protein (PrP), show a major diversity in nomadism , 40. Thi6Ser in tPRNP characterization also in sheep populations of the same areas, especially because mixed holdings are very common. A surveillance plan for the disease in both goats and sheep populations would be also important to understand the true epidemiology of scrapie in the country. The occurrence of a new prion disease in dromedaries, CPD in Algeria, could be an impetus for a surveillance plan for prion diseases in Africa since dromedaries are often bred with sheep and goats simultaneously; sharing common pastures in many African countries such as in Ethiopia. A new prion disease in a farmed animal species requires a thorough risk assessment for implementing policies to control the disease in animals and minimize human exposure [PRNP polymorphisms of goats in economically marginal areas and increase the interest in exploring the genetic variation of the PRNP gene in native goat breeds and sheep as well, in countries where cross-breeding with imported animals has never been performed.The two novel amino acid substitutions and the high frequency of the scrapie protective variant p.Asn146Ser) might indicate a lower risk of scrapie in Ethiopian goats. It could be very important to perform 46Ser migPRNP gene: Western and Central Highlands and Long-eared Somali. Somatic aspects of the different goats have been already described [n = 52) were sampled in the Seka, Tiroafata and Omonada districts of Jimma; Western Highland (n = 110) from the Ambo and Bako districts; and Long eared Somali (n = 67) from the Gursum and Jijiga districts of Eastern Hararghe. The samples consisted of 75% males and 25% females with no genetic relation.Three of the main Ethiopian native goat breeds were analysed for the escribed , 16, 41.PRNP (NM_001314247.1 for the cDNA position and NP_001301176.1 for the protein variants) was performed and direct PCR fragment sequencing was carried out as previously described [Polymerase chain reaction (PCR) amplification of the open reading frame of the caprine Raw data of Ethiopian goats genotyping are reported in Additional file 1Amino acid variants are described following recommendation of Den Dunnen et al. Allelic frequencies were calculated using the Hardy-Weinberg equation. Statistical differences between the breeds were calculated with a chi-square test.Additional file 1:Prion protein gene polymorphism in native Ethiopian goats. Excel file that include all information related to each single goat in the three Ethiopian regions. For each goat, information regarding sex, age in months and the related prion polymorphisms are reported. (XLS 60 kb)Additional file 2:Table of the genotypes combinations of Prion protein gene in native Ethiopian goats.. (PDF 423 kb)


Community engagement is increasingly recognized as a critical element of medical research, recommended by ethicists, required by research funders and advocated in ethics guidelines. The benefits of community engagement are often stressed in instrumental terms, particularly with regard to promoting recruitment and retention in studies. Less emphasis has been placed on the value of community engagement with regard to ethical good practice, with goals often implied rather than clearly articulated. This article outlines explicitly how community engagement can contribute to ethical global health research by complementing existing established requirements such as informed consent and independent ethics review. The overarching and interlinked areas are (1) respecting individuals, communities and stakeholders; (2) building trust and social relationships; (3) determining appropriate benefits; minimizing risks, burdens and exploitation; (4) supporting the consent process; (5) understanding vulnerabilities and researcher obligations; (6) gaining permissions, approvals and building legitimacy and (7) achieving recruitment and retention targets. Engagement therefore denotes a wide variety of activities, such as patient and public involvement events (e.g. science cafes events), consultation regarding study activities through community advisory boards . Informing and explaining a study to communities requires providing information about the researcher, the research institutions, the study concept and rationale in appropriate formats  involved  health care can help to build institutional trust  providing potential participants with adequate information about the proposed study; (2) potential participants understanding what is proposed and (3) participants being able to make a voluntary decision to participate , Although community engagement can help identify specific vulnerabilities, there can be inadvertent consequences of community engagement  that are difficult to reconcile with the limits of the protocol Lavery, . CommuniWhen seeking formal permissions, approvals and building legitimacy, focused and in-depth engagement is needed. One option is a deliberative approach that entails reaching a consensus while acknowledging differing views  respecting individuals, communities and stakeholders; (2) building trust and social relationships; (3) determining appropriate benefits; minimizing risks, burdens and exploitation; (4) supporting the consent process; (5) understanding vulnerabilities and researcher obligations; (6) gaining permissions, approvals and building legitimacy and (7) achieving recruitment and retention targets.


In January 2016, highly pathogenic avian influenza (HPAI) A(H7N8) virus and low pathogenicity avian influenza (LPAI) A(H7N8) virus were detected in commercial turkey flocks in Dubois County, Indiana. The Indiana State Department of Health (ISDH) and the Dubois County Health Department (DCHD) coordinated the public health response to this outbreak, which was the first detection of HPAI A(H7N8) in any species ( On January 15, 2016, ISDH was notified by the Indiana State Board of Animal Health that HPAI A(H7N8) virus had been confirmed in a commercial turkey flock in Dubois County, Indiana. In accordance with USDA guidelines (ISDH and DCHD recommended that responders be monitored during the response and for 10 days after their last possible exposures for influenza-like illness (ILI), defined as either 1) self-reported fever with cough or sore throat, or 2) conjunctivitis with or without additional symptoms. All responders received instructions to seek medical attention and contact public health authorities if they developed ILI during the 10-day period. USDA monitored federal employees, contractors, and subcontractors who participated in the response. ISDH and DCHD monitored state and local responders, using adapted, unpublished USDA/CDC public health monitoring procedures for HPAI A(H5) that were first circulated in September 2015 and later updated in November 2015. Responders were classified into three risk categories: 1) no risk, 2) low but not zero risk, and 3) some risk. Responders with low but not zero risk were those exposed to infected or potentially infected birds or their environments while using appropriate personal protective equipment (PPE)  were monitored for some risk exposures, 67 (40%) were monitored for low but not zero risk exposures, four (2%) were monitored with no risk status recorded, seven (4%) had no exposure, five (3%) declined to be monitored, and nine (5%) were lost to follow-up. Among the 145 persons who were monitored, 14 (10%) reported current or recent ILI symptoms, 12 (86%) of whom were tested during January 16–22, 2016, with a median of 1 day from onset to medical evaluation. Nine patients had nasopharyngeal/oropharyngeal specimens collected, one had a conjunctival specimen collected, and two had both types of specimens collected. Specimens were tested for influenza A virus by real-time reverse transcription–polymerase chain reaction at the ISDH Laboratories; all 12 patients tested negative for influenza A virus.On January 15, 2016, Indiana activated its Emergency Operations Center with staffing consistent with the Federal Emergency Management Agency Emergency Support Functions (ESFs) appropriate to the response .ISDH’s major actions during the public health response included developing a demobilization packet for responders, issuing a Health Alert Network advisory to Dubois County and surrounding counties with recommendations for health care providers, establishing syndromic surveillance queries in the Indiana Public Health Emergency Surveillance System to detect community-acquired cases, and developing recommendations for the use of antivirals.Public health monitoring procedures for H7N8 responders were successfully implemented during this outbreak; no cases of zoonotic avian influenza infection were detected. The risk for zoonotic transmission in this outbreak was thought to be low at the time. No human infections with influenza A(H7N8) viruses had ever been reported,The unpublished USDA/CDC public health monitoring procedures that were adapted for use in this outbreak were developed for responders to an HPAI A(H5) outbreak. HPAI and LPAI are differentiated based on genetic features and the extent to which these viruses produce morbidity and mortality in poultry . The updated plan will allow for passive monitoring of persons wearing PPE and responding to certain influenza A H5 and H7 viruses that have caused outbreaks in the United States but have no history of causing human infections . The updated plan will also cover all avian influenza viruses of public health concern, including both HPAI and LPAI viruses.Several challenges to human health monitoring were identified during this outbreak. First, receipt of contact information for responders by the local health department was delayed in the initial stages because of the urgency and complexity of the animal health response. Complete information for exposed responders was not received by DCHD until 5 days into the response (on January 20), although preliminary information was provided earlier. Second, tears in Tyvek suits were reportedly common because of the nature of animal handling activities; this could have resulted in misclassification of disease exposure risk. Finally, mobilization of a large number of responders within a short period raised concerns that PPE and monitoring recommendations were not being implemented consistently.Enhanced communication and information sharing among local, state, and federal agencies would improve identification of exposed persons and coordination of specimen collection, testing, and medical care for ill responders. This could be accomplished by 1) effectively communicating public health needs and recommendations to all stakeholders in the response; 2) identifying processes for early identification of exposed persons ; 3) designating a local/state/federal public health department liaison to be embedded in the unified command post to facilitate coordinated implementation of human health monitoring, including obtaining names and contact information for responders; and 4) conducting daily debriefings with safety officers in the incident command system to identify injuries or breaches in PPE that could elevate responder risk. In future outbreaks, ISDH will also recommend that responders to outbreaks of AI viruses of public health concern entering the exclusion zone (hot zone) and contamination reduction zone (warm zone) in infected premises (Prolonged or close contact with birds infected with avian influenza (AI) virus increases the risk for zoonotic infection in humans. Monitoring exposed persons for 10 days might facilitate early detection and reporting of zoonotic AI.Monitoring procedures for highly pathogenic AI (HPAI) responders were successfully implemented during a 2016 outbreak of HPAI A(H7N8) in commercial turkey flocks in Indiana. No human cases of AI were identified.Collaboration among local, state, and federal partners is essential during AI outbreak responses. Monitoring should be considered for all responders who had contact with infected birds or their environments, regardless of whether personal protective equipment was worn.


In November 2011, a government hospital physician in Shibganj sub-district of Bangladesh reported a cluster of patients with fever and joint pain or rash. A multi-disciplinary team investigated to characterize the outbreak; confirm the cause; and recommend control and prevention measures.Shibganj's residents with new onset of fever and joint pain or rash between 1 September and 15 December 2011 were defined as chikungunya fever (CHIKF) suspect cases. To estimate the attack rate, we identified 16 outpatient clinics in 16 selected wards across 16 unions in Shibganj and searched for suspect cases in the 80 households nearest to each outpatient clinic. One suspect case from the first 30 households in each ward was invited to visit the nearest outpatient clinic for clinical assessment and to provide a blood sample for laboratory testing and analyses.We identified 1,769 CHIKF suspect cases from among 5,902 residents surveyed (30%). Their median age was 28 (IQR:15−42) years. The average attack rate in the sub-district was 30% (95% CI: 27%−33%). The lowest attack rate was found in children <5 years (15%). Anti-CHIKV IgM antibodies were detected by ELISA in 78% (264) of the 338 case samples tested. In addition to fever, predominant symptoms of serologically-confirmed cases included joint pain (97%), weakness (54%), myalgia (47%), rash (42%), itching (37%) and malaise (31%). Among the sero-positive patients, 79% (209/264) sought healthcare from outpatient clinics. CHIKV was isolated from two cases and phylogenetic analyses of full genome sequences placed these viruses within the Indian Ocean Lineage (IOL). Molecular analysis identified mutations in E2 and E1 glycoproteins and contained the E1 A226V point mutation.The consistently high attack rate by age groups suggested recent introduction of chikungunya in this community. Mosquito control efforts should be enhanced to reduce the risk of continued transmission and to improve global health security. Togaviridae family, have been reported from several countries in Africa, Asia and the Western Pacific since 1952.[Aedes aegypti and Aedes albopictus mosquitoes are the two primary vectors transmitting CHIKV to humans.[Epidemics of chikungunya fever (CHIKF) caused by chikungunya virus (CHIKV), a mosquito-transmitted alpha virus of the nce 1952. Large ounce 1952.–12 Aedeso humans. Classicao humans.–16 Howevo humans., 17–19 Fo humans.–30During December 2008, CHIKV infection was confirmed for the first time in Bangladesh. The outbOn 5 November 2011, the primary healthcare physician managing the government hospital in Shibganj sub-district of Chapainababganj district in northwestern Bangladesh reported a cluster of 19 patients with fever, severe joint pain and/or rash to the Institute of Epidemiology, Disease Control and Research (IEDCR) of the Ministry of Health and Family Welfare. In response, a collaborative, multi-disciplinary investigation team from IEDCR and icddr,b conducted an investigation to (i) characterize the outbreak in terms of person, place and time; (ii) confirm the cause; and (iii) identify prevention and control approaches for future outbreaks.The team visited the hospital in the affected sub-district and conducted unstructured interviews with local healthcare providers, including physicians and field workers, to collect preliminary information about the clinical features, laboratory findings, and geographical distribution of affected patients. Based on these interviews, the experience of a similar outbreak in 2008, and giveGiven the lack of information regarding the prevalence of chikungunya fever from Bangladesh during the investigation period in 2011 and because the preliminary survey findings suggested a widespread distribution of suspect cases, the investigation team conducted a house-to-house syndromic survey in all of the 16 unions of Shibganj to calculate the attack rate. Using the attack rate of 34% as observed during the preliminary survey and assuming a type 1 error of 0.05, a precision of 5%, an average of 5 persons per household, a design effect of 3.0 to take into account the cluster sampling design and a non-response rate of 20% at the household level, we calculated a required sample size of at least 1,280 households from the 16 unions of Shibganj for the syndromic survey. In each To obtain clinical histories and collect biological specimens, data collectors invited one suspect case from each of the first 30 households visited in each ward to come to the nearest primary healthcare clinic to complete a clinical assessment and to provide a blood sample for testing. If a household had more than one eligible respondent, the data collectors requested the eldest household member present during the survey to select one resident for participation. Physicians on the team later interviewed and physically examined suspect cases who came to the local clinics.To test for IgM antibodies to CHIKV, trained phlebotomists collected 5 ml venous blood from each suspect case who had presented to the clinic and consented to participate. Phlebotomists stored all blood samples immediately after collection at 4−8°C. The samples were centrifuged on the same day and the sera were stored in the local hospital refrigerator and later transported in cool boxes to the Virology Laboratory of IEDCR. At IEDCR, each serum sample was divided into three aliquots before storing at -70°C until testing. At the beginning of the investigation, we only had 300 rapid test kits available, so we tested 300 randomly selected samples out of the 367 sera. We tested 338 randomly selected aliquots out of the 367 samples for anti-CHIKV immunoglobulin M (IgM) antibodies using an enzyme-linked immunosorbent assay (ELISA) test kit that was also available with our group during the investigation  in the Virology Laboratory of IEDCR following the manufacturer’s guidelines.Acute sera that had been collected from suspect cases within two days of illness onset and tested negative for IgM antibodies to CHIKV were later tested for viral RNA using quantitative reverse transcription polymerase chain reaction (RT-qPCR). The seraPhylogenetic analysis was performed as previously described. ChikunguAedes aegypti bred mainly in the water collected in artificial containers in the environment of human settlements in Asia,[Aedes albopictus bred predominantly in natural containers including coconut shells, leaf axils and tree holes, team entomologists looked for mosquito larvae in both man-made and natural water containers.[The entomological investigation was conducted during December 15−31, 2011 and focused on identifying vector species. Given that  in Asia, while Aentainers. To collentainers.We divided the total number of suspect cases by the total number of household residents surveyed in each cluster ward within each union composed of the 80 households nearest to an outpatient clinic in the ward to calculate the attack rates by cluster. We estimated the attack rate within the sub-district by calculating the mean of attack rates for the 16 clusters combined. We also calculated the attack rates by 5-year age groups and gender. We used the number of suspect cases instead of confirmed cases to estimate the attack rate because we only collected and tested the serum samples from a sub-set of suspect cases who later came to clinics for clinical assessment and sample provision. For the sub-set of patients who came to the clinics for clinical evaluation, we described their clinical features. Since many respondents were still symptomatic during the clinical survey, we estimated the median follow-up time from symptom onset till recovery for each of the four symptoms including joint pain, joint swelling, debilitating weakness and severe myalgia. Previous authors have suggested that length of follow-up time should be extracted from the time frame where the Kaplan Meier estimate was most stable. We consiThe Institutional Review Board (IRB) of IEDCR and the Ethical Review Committee (ERC) of icddr,b considers outbreak investigations that primarily aim to control outbreaks as public health practice and approval by research ethics committee is not required. This is in line with national and international ethical guidelines including the 1991 International Guidelines for Ethical Review of Epidemiological Studies by the Council for International Organizations of Medical Sciences.–49 SeverWe obtained informed oral consent for voluntary participation from adults and verbal assent from children. Past experience of research in similar settings suggested that obtaining written informed consent was difficult, time consuming and resource-intensive when dealing with illiterate participants who were often very cautious, as they did not know what they were signing or whether the signed document could be used against them. As a result, verbal informed consent procedure was allowed during this emergency outbreak situation, where the majority of the affected population were illiterate and ill, as obtaining written informed consent was considered unnecessary and not in the patient and community’s best interest. The investigation team member who was informing a participant signed a form stating that the appropriate information was provided and verbal consent received before beginning any information and/or sample collection or any kind of investigation.The team identified 30 suspect cases from the hospital initially. These cases resided in six different unions within Shibganj. Their median age was 35  years and 45% were males. Fever (100%), polyarthralgia (90%), malaise (83%), severe weakness (66%), rash (48%), itching (38%), and headache (35%) were the predominant symptoms. Forty percent (12/30) of these patients had evidence of IgM CHIKV antibodies in their sera when tested using the rapid assay.We surveyed a total of 5,902 individuals from 1,283 households in the 16 unions of Shibganj. Of these, 1,769 (30%) met the suspect case definition. The median age of suspect cases was 28 (IQR:15−42) years. The average attack rate in the sub-district was 30% (95% CI: 27%−33%). Attack rates varied from 10%−44% across the 16 selected clusters within the 16 unions of Shibganj . The meaAmong the 1,769 suspect cases, 480 (27%) were invited to participate in the clinical survey and to provide blood samples. Seventy-nine percent (377/480) of the invited suspect cases gave consent for the clinical survey and physical examination, and 367 (77%) provided a blood sample. Among the serum samples tested with the rapid test, 73% were positive for chikungunya whereas IgM anti-CHIKV antibodies were detected in 78% (264/338) of the samples that were tested by ELISA. We found that the suspect cases tested within one week of illness onset (43%) were less likely to have IgM antibodies against Chikungunya compared to the suspect cases tested more than 60 days post illness onset (98%) .Among the 377 cases interviewed during the clinical survey, the median age was 40 years (IQR:30−50 years); 64% (241/377) were females; and 92% (347/377) were Muslims. The main clinical symptoms included fever (100%), joint pain (88%), debilitating weakness preventing routine activities (55%), rash (51%), severe myalgia (44%), itching (33%) and joint swelling (16%) . Thirty-In addition to fever, the predominant symptoms of serologically-confirmed cases (n = 264) among those who had provided sera after coming to the clinic included joint pain (97%), weakness (54%), myalgia (47%), rash (42%), itching (37%) and malaise (31%). The rashes were found on the trunk (36%), lower limbs (36%), palms and soles (33%), upper limbs in (14%), and face (8%). Of those who developed joint pain, 93% (238/256) reported pain in multiple joints and joints of the lower limbs were involved in 90% (230/256) of the cases. Among the cases reporting joint pain, severe pain that prevented sleep was reported by 61% (156/256), moderate pain that caused pain on movement was reported by 23% (59/256) and minor pain that did not interfere with daily activities but caused discomfort was reported by 17% (43/256). Severe weakness was reported by 40% (105/264), debilitating myalgia by 36% (94/264) and severe joint swelling by 14% (37/264). On examination, team physicians found restricted joint mobility in 23% (59/256) of the serologically confirmed cases. The rashes were maculopapular in 62% (69/112), macular in 33% (37/112) and popular in 5% (6/112) of the suspect cases with rashes. The median durations of symptom persistence of joint pain was 31 days, while joint swelling persisted for 7 days, debilitating weakness and severe myalgia, each persisted for about 12 days in the majority of this sub-set of confirmed cases . FurtherTwo out of the four patients from this outbreak in Shibganj and the single patient from the concurrent outbreak in Dohar sub-district of Dhaka whose acute sera were collected within two days of illness onset had detectable Chikungunya virus RNA with RT-qPCR. However, none of them had IgM antibodies in their sera. The three viral isolates, obtained from the two outbreaks including this outbreak in Shibganj (n = 2) and in Dohar (n = 1), clustered with strains from the 2006 Indian Ocean lineage and were most closely related to the Indian/Asian geographic isolates rather than the African isolates within that lineage . The sizAedes aegypti or Aedes albopictus mosquitoes during the investigation. The team found mosquito larva from only 1/160 households surveyed and the larvae that hatched from this household in IEDCR’s Entomology laboratory yielded Culex quinquefaciatus species.The team did not find any larvae of Aedes aegypti or Aedes albopictus mosquitoes during the investigation.We identified CHIKF suspect cases among 30% of the residents surveyed across the 16 unions in Shibganj during the syndromic survey. The median age of suspect cases was 28 years. The lowest attack rate was found in children <5 years (15%). Anti-chikungunya virus (CHIKV) IgM antibodies were detected by an enzyme-linked immunosorbent assay (ELISA) in 78% of the case samples tested. In addition to fever, the predominant symptoms of serologically-confirmed cases, included joint pain (97%), weakness (54%), myalgia (47%), rash (42%), itching (37%) and malaise (31%). The median durations of symptom persistence of severe joint pain was 31 days, while severe joint swelling persisted for 7 days, debilitating weakness and severe myalgia, each persisted for about 12 days in the majority of this sub-set of confirmed cases. Among the sero-positive patients, 79% sought healthcare from outpatient clinics. CHIKV was isolated from two cases and phylogenetic analyses of full genome sequences placed these viruses within the Indian Ocean Lineage (IOL). Molecular analysis identified mutations in E2 and E1 glycoproteins and contained the E1 A226V point mutation. We did not find any larvae of The clinical syndrome of fever, multiple joint pain, with or without debilitating weakness or severe myalgia was consistent with previous CHIKF outbreaks., 5, 33 TThough several outbreaks have reported an increased propensity to cause clinical illness among females, our syndAedes albopictus larvae breeding in the numerous wet earthen and clay pots that were used for pottery.[Aedes mosquito from the affected area, a finding quite inconsistent with past outbreaks. The delayed reporting of the outbreak with the consequent delay in conducting the investigation during the dry winter months of November and December possibly prevented us from detecting the vector. Given the role of environmental variables on Aedes albopictus and on the pathogen transmissibility, we believe that transmission likely stopped due to changes in weather that constrained Aedes mosquito populations during this outbreak.[Aedes larvae by December, and other outbreaks in Bangladesh have also stopped abruptly around the months of November and December.[Aedes albopictus species.[Aedes albopictus were most likely responsible for transmission,[The first recognized outbreak in Chapainababganj district in 2008 predominantly affected Hindu potters and had limited geographic spread. The outbreak was reportedly precipitated by the high concentration of  pottery. During t pottery., 14, 56  pottery., 33 despoutbreak., 58 We wDecember., 33, 34  species. Another A major limitation of our investigation was that the suspect cases that presented for clinical assessment and blood collection were unlikely representative of the entire affected population. We expect that cases with severe and/or persistent symptoms were more likely to come to the clinic compared to milder cases or patients who had recovered, so our clinical descriptions of serologically confirmed cases were likely biased towards severe presentations. However, since almost half of the cases had already recovered from clinical illness during the clinical survey, we also captured some less serious disease. Given that an evaluation of the sensitivity and specificity of clinical criteria for CHIKV infection identified the syndrome combining fever and polyarthralgias to allow for the correct classification of 87% of those who had serologically confirmed CHIKV infections, our caseAedes populations.Though no recent community seroprevalence study of CHIKV has been published from Bangladesh, a post outbreak sero-survey conducted in selected outbreak affected communities under publication by the author group as well as a 1995 cross-sectional survey carried out in the neighbouring city of Calcutta, India indicated that the level of previous exposure to Chikungunya infection in the country remained low. FurthermAedes mosquitoes in the future may help provide important insight into emergence of epidemic strains in Bangladesh. Effective mosquito control using a comprehensive and integrated pest management approach targeted at every life stage of a mosquito could reduce the risk of CHIKV dissemination.[Population-based seroprevalence and risk factor surveys could help describe the true extent of previous circulation of this virus and identify factors that might put some communities at higher risk than others for outbreaks. Molecular clock analyses and monitoring the spread of genotype viruses and the proportion of the mination., 13 To eS1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S1 Fig(DOCX)Click here for additional data file.S1 Text(DOCX)Click here for additional data file.S1 Dataset(SAV)Click here for additional data file.S2 Dataset(SAV)Click here for additional data file.S3 Dataset(SAV)Click here for additional data file.