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BACKGROUND: In cattle, genome-wide association studies (GWAS) have largely focused on European or Asian breeds, using genotyping arrays that were primarily designed for European cattle. Because there is growing interest in performing GWAS in African breeds, we have assessed the performance of 23 commercial bovine genotyping arrays for capturing the diversity across African breeds and performing imputation. We used 409 whole-genome sequences (WGS) spanning global cattle breeds, and a real cohort of 2481 individuals (including African breeds) that were genotyped with the Illumina high-density (HD) array and the GeneSeek bovine 50 k array. RESULTS: We found that commercially available arrays were not effective in capturing variants that segregate among African indicine animals. Only 6% of these variants in high linkage disequilibrium (LD) (r2 > 0.8) were on the best performing arrays, which contrasts with the 17% and 25% in African and European taurine cattle, respectively. However, imputation from available HD arrays can successfully capture most variants (accuracies up to 0.93), mainly when using a global, not continent-specific, reference panel, which partially reflects the unusually high levels of admixture on the continent. When considering functional variants, the GGPF250 array performed best for tagging WGS variants and imputation. Finally, we show that imputation from low-density arrays can perform almost as well as HD arrays, if a two-stage imputation approach is adopted, i.e. first imputing to HD and then to WGS, which can potentially reduce the costs of GWAS. CONCLUSIONS: Our results show that the choice of an array should be based on a balance between the objective of the study and the breed/population considered, with the HD and BOS1 arrays being the best choice for both taurine and indicine breeds when performing GWAS, and the GGPF250 being preferable for fine-mapping studies. Moreover, our results suggest that there is no advantage to using the indicus-specific arrays for indicus breeds, regardless of the objective. Finally, we show that using a reference panel that better represents global bovine diversity improves imputation accuracy, particularly for non-European taurine populations.
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Estudio de Asociación del Genoma Completo , Polimorfismo de Nucleótido Simple , Animales , Bovinos/genética , Genotipo , Desequilibrio de LigamientoRESUMEN
BACKGROUND: Maize lethal necrosis is caused by a synergistic co-infection of Maize chlorotic mottle virus (MCMV) and a specific member of the Potyviridae, such as Sugarcane mosaic virus (SCMV), Wheat streak mosaic virus (WSMV) or Johnson grass mosaic virus (JGMV). Typical maize lethal necrosis symptoms include severe yellowing and leaf drying from the edges. In Kenya, we detected plants showing typical and atypical symptoms. Both groups of plants often tested negative for SCMV by ELISA. METHODS: We used next-generation sequencing to identify viruses associated to maize lethal necrosis in Kenya through a metagenomics analysis. Symptomatic and asymptomatic leaf samples were collected from maize and sorghum representing sixteen counties. RESULTS: Complete and partial genomes were assembled for MCMV, SCMV, Maize streak virus (MSV) and Maize yellow dwarf virus-RMV (MYDV-RMV). These four viruses (MCMV, SCMV, MSV and MYDV-RMV) were found together in 30 of 68 samples. A geographic analysis showed that these viruses are widely distributed in Kenya. Phylogenetic analyses of nucleotide sequences showed that MCMV, MYDV-RMV and MSV are similar to isolates from East Africa and other parts of the world. Single nucleotide polymorphism, nucleotide and polyprotein sequence alignments identified three genetically distinct groups of SCMV in Kenya. Variation mapped to sequences at the border of NIb and the coat protein. Partial genome sequences were obtained for other four potyviruses and one polerovirus. CONCLUSION: Our results uncover the complexity of the maize lethal necrosis epidemic in Kenya. MCMV, SCMV, MSV and MYDV-RMV are widely distributed and infect both maize and sorghum. SCMV population in Kenya is diverse and consists of numerous strains that are genetically different to isolates from other parts of the world. Several potyviruses, and possibly poleroviruses, are also involved.
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Gammaherpesvirinae/genética , Genoma Viral , Luteoviridae/genética , Potyviridae/genética , Potyvirus/genética , Zea mays/virología , Secuencia de Aminoácidos , Proteínas de la Cápside/genética , Mapeo Cromosómico , Gammaherpesvirinae/clasificación , Gammaherpesvirinae/aislamiento & purificación , Gammaherpesvirinae/patogenicidad , Secuenciación de Nucleótidos de Alto Rendimiento , Kenia , Luteoviridae/clasificación , Luteoviridae/aislamiento & purificación , Luteoviridae/patogenicidad , Metagenómica/métodos , Filogenia , Enfermedades de las Plantas/virología , Hojas de la Planta/virología , Polimorfismo Genético , Potyviridae/clasificación , Potyviridae/aislamiento & purificación , Potyviridae/patogenicidad , Potyvirus/clasificación , Potyvirus/aislamiento & purificación , Potyvirus/patogenicidad , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Sorghum/virologíaRESUMEN
OBJECTIVE: This study was carried out to assess the haplotype diversity and population dynamics in cattle populations of Ethiopia. METHODS: We sequenced the complete mitochondrial cytochrome b gene of 76 animals from five indigenous and one Holstein Friesian×Barka cross bred cattle populations. RESULTS: In the sequence analysis, 18 haplotypes were generated from 18 segregating sites and the average haplotype and nucleotide diversities were 0.7540±0.043 and 0.0010±0.000, respectively. The population differentiation analysis shows a weak population structure (4.55%) among the populations studied. Majority of the variation (95.45%) is observed by within populations. The overall average pair-wise distance (FST) was 0.049539 with the highest (FST = 0.1245) and the lowest (FST = 0.011) FST distances observed between Boran and Abigar, and Sheko and Abigar from the indigenous cattle, respectively. The phylogenetic network analysis revealed that all the haplotypes detected clustered together with the Bos taurus cattle and converged to a haplogroup. No haplotype in Ethiopian cattle was observed clustered with the reference Bos indicus group. The mismatch distribution analysis indicates a single population expansion event among the cattle populations. CONCLUSION: Overall, high haplotype variability was observed among Ethiopian cattle populations and they share a common ancestor with Bos taurus.
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BACKGROUND: Aphids are major vectors of plant viruses. Common bean (Phaseolus vulgaris L.) and maize (Zea mays L.) are important crops that are vulnerable to aphid herbivory and aphid-transmitted viruses. In East and Central Africa, common bean is frequently intercropped by smallholder farmers to provide fixed nitrogen for cultivation of starch crops such as maize. We used a PCR-based technique to identify aphids prevalent in smallholder bean farms and next generation sequencing shotgun metagenomics to examine the diversity of viruses present in aphids and in maize leaf samples. Samples were collected from farms in Kenya in a range of agro-ecological zones. RESULTS: Cytochrome oxidase 1 (CO1) gene sequencing showed that Aphis fabae was the sole aphid species present in bean plots in the farms visited. Sequencing of total RNA from aphids using the Illumina platform detected three dicistroviruses. Maize leaf RNA was also analysed. Identification of Aphid lethal paralysis virus (ALPV), Rhopalosiphum padi virus (RhPV), and a novel Big Sioux River virus (BSRV)-like dicistrovirus in aphid and maize samples was confirmed using reverse transcription-polymerase chain reactions and sequencing of amplified DNA products. Phylogenetic, nucleotide and protein sequence analyses of eight ALPV genomes revealed evidence of intra-species recombination, with the data suggesting there may be two ALPV lineages. Analysis of BSRV-like virus genomic RNA sequences revealed features that are consistent with other dicistroviruses and that it is phylogenetically closely related to dicistroviruses of the genus Cripavirus. CONCLUSIONS: The discovery of ALPV and RhPV in aphids and maize further demonstrates the broad occurrence of these dicistroviruses. Dicistroviruses are remarkable in that they use plants as reservoirs that facilitate infection of their insect replicative hosts, such as aphids. This is the first report of these viruses being isolated from either organism. The BSRV-like sequences represent a potentially novel dicistrovirus infecting A. fabae.
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Áfidos/virología , Dicistroviridae/clasificación , Dicistroviridae/genética , Granjas , Metagenoma , Phaseolus/parasitología , Zea mays/parasitología , Animales , Kenia , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADNRESUMEN
BACKGROUND: The advent of genotyping by Next Generation Sequencing has enabled rapid discovery of thousands of single nucleotide polymorphism (SNP) markers and high throughput genotyping of large populations at an affordable cost. Genotyping by sequencing (GBS), a reduced representation library sequencing method, allows highly multiplexed sequencing of genomic subsets. This method has limitations for small organisms with low amounts of genomic DNA, such as the bluetongue virus (BTV) vectors, Culicoides midges. RESULTS: This study employed the GBS method to isolate SNP markers de novo from whole genome amplified Culicoides brevitarsis genomic DNA. The individuals were collected from regions representing two different Australian patterns of BTV strain distribution: the Northern Territory (NT) and the east coast. We isolated 8145 SNPs using GBS. Phylogenetic analysis conducted using the filtered 3263 SNPs revealed the presence of a distinct C. brevitarsis sub-population in the NT and this was confirmed by analysis of mitochondrial DNA. Two loci showed a very strong signal for selection and were unique to the NT population. Bayesian analysis with STRUCTURE indicated a possible two-population cluster. CONCLUSIONS: The results suggest that genotyping vectors with high density markers in combination with biological and environmental data is useful. However, more extensive sampling over a wider spatial and temporal range is needed. The presence of sub-structure in populations and loci under natural selection indicates the need for further investigation of the role of vectors in shaping the two Australian systems of BTV transmission. The described workflow is transferable to genotyping of small, non-model organisms, including arthropod vectors of pathogens of economic and medical importance.
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Ceratopogonidae/genética , Genética de Población , Genoma de los Insectos , Genómica , Genotipo , Animales , Teorema de Bayes , Complejo IV de Transporte de Electrones/genética , Biblioteca de Genes , Genes Mitocondriales , Genómica/métodos , Haplotipos , Northern Territory , Filogenia , Polimorfismo de Nucleótido Simple , Sitios de Carácter Cuantitativo , Selección GenéticaRESUMEN
Noroviruses (NoVs) and sapoviruses (SaVs), which belong to the family Caliciviridae, are important human and animal enteric pathogens with zoonotic potential. In Ethiopia, no study has been done on the epidemiology of animal NoVs and SaVs. The aim of this study was to detect and characterize NoVs and SaVs from swine of various ages. Swine fecal samples (n = 117) were collected from commercial farms in Ethiopia. The samples were screened for caliciviruses by reverse transcription polymerase chain reaction (RT-PCR) using universal and genogroup-specific primer pairs. Phylogenetic analysis was conducted using a portion of the RNA-dependent RNA polymerase (RdRp) region and the VP1 region of genome sequences of caliciviruses. Among 117 samples, potential caliciviruses were detected by RT-PCR in 17 samples (14.5 %). Of the RT-PCR-positive fecal samples, four were sequenced, of which two were identified as human NoV GII.1 and the other two as porcine SaV GIII. The porcine SaV strains that were detected were genetically related to the porcine enteric calicivirus Cowden strain genogroup III (GIII), which is the prototype porcine SaV strain. No porcine NoVs were detected. Our results showed the presence of NoVs in swine that are most similar to human strains. These findings have important implications for NoV epidemiology and food safety. Therefore, continued surveillance of NoVs in swine is needed to define their zoonotic potential, epidemiology and public and animal health impact. This is the first study to investigate enteric caliciviruses (noroviruses and sapoviruses) in swine in Ethiopia.
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Infecciones por Caliciviridae/veterinaria , Norovirus/clasificación , Norovirus/aislamiento & purificación , Sapovirus/clasificación , Sapovirus/aislamiento & purificación , Enfermedades de los Porcinos/virología , Animales , Infecciones por Caliciviridae/virología , Análisis por Conglomerados , Etiopía , Heces/virología , Norovirus/genética , Filogenia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sapovirus/genética , Análisis de Secuencia de ADN , Homología de Secuencia , Porcinos , Proteínas Virales/genéticaRESUMEN
Viral gastroenteritis is a major public health problem worldwide. In Ethiopia, very limited studies have been done on the epidemiology of enteropathogenic viruses. The aim of this study was to detect and characterize noroviruses (NoVs) and sapoviruses (SaVs) from acute gastroenteritis patients of all ages. Fecal samples were collected from diarrheic patients (n = 213) in five different health centers in Addis Ababa during June-September 2013. The samples were screened for caliciviruses by reverse transcription polymerase chain reaction (RT-PCR) using universal and genogroup-specific primer pairs. Phylogenetic analyses were conducted using the sequences of the PCR products. Of the clinical samples, 25.3 % and 4.2 % were positive for NoV and SaV RNA, respectively. Among the norovirus positives, 22 were sequenced further, and diverse norovirus strains were identified: GI (n = 4), GII (n = 17) and GIV (n = 1). Most strains were GII (n = 17/22: 77.2 %), which were further divided into three different genotypes (GII.4, GII.12/GII.g recombinant-like and GII.17), with GII.17 being the dominant (7/17) strain detected. GI noroviruses, in particular GI.4 (n = 1), GI.5 (n = 2) and GI.8 (n = 1), were also detected and characterized. The GIV strain detected is the first from East Africa. The sapoviruses sequenced were also the first reported from Ethiopia. Collectively, this study showed the high burden and diversity of noroviruses and circulation of sapoviruses in diarrheic patients in Ethiopia. Continued surveillance to assess their association with diarrhea is needed to define their epidemiology, disease burden, and impact on public health.
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Infecciones por Caliciviridae/virología , Gastroenteritis/virología , Norovirus/aislamiento & purificación , Sapovirus/aislamiento & purificación , Adolescente , Adulto , Anciano , Infecciones por Caliciviridae/epidemiología , Niño , Preescolar , Diarrea/epidemiología , Diarrea/virología , Etiopía/epidemiología , Heces/virología , Femenino , Gastroenteritis/epidemiología , Genotipo , Humanos , Lactante , Masculino , Persona de Mediana Edad , Norovirus/clasificación , Norovirus/genética , Filogenia , Prevalencia , Sapovirus/clasificación , Sapovirus/genética , Adulto JovenRESUMEN
Bluetongue virus (BTV) is a major pathogen of ruminants that is transmitted by biting midges (Culicoides spp.). Australian BTV serotypes have origins in Asia and are distributed across the continent into two distinct episystems, one in the north and another in the east. Culicoides brevitarsis is the major vector of BTV in Australia and is distributed across the entire geographic range of the virus. Here, we describe the isolation and use of DNA microsatellites and gauge their ability to determine population genetic connectivity of C. brevitarsis within Australia and with countries to the north. Eleven DNA microsatellite markers were isolated using a novel genomic enrichment method and identified as useful for genetic analyses of sampled populations in Australia, northern Papua New Guinea (PNG) and Timor-Leste. Significant (P < 0.05) population genetic subdivision was observed between all paired regions, though the highest levels of genetic sub-division involved pair-wise tests with PNG (PNG vs. Australia (FST = 0.120) and PNG vs. Timor-Leste (FST = 0.095)). Analysis of multi-locus allelic distributions using STRUCTURE identified a most probable two-cluster population model, which separated PNG specimens from a cluster containing specimens from Timor-Leste and Australia. The source of incursions of this species in Australia is more likely to be Timor-Leste than PNG. Future incursions of BTV positive C. brevitarsis into Australia may be genetically identified to their source populations using these microsatellite loci. The vector's panmictic genetic structure within Australia cannot explain the differential geographic distribution of BTV serotypes.
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Ceratopogonidae/genética , ADN/genética , Variación Genética , Genética de Población/métodos , Insectos Vectores/genética , Repeticiones de Microsatélite , Animales , Australia , Virus de la Lengua Azul/fisiología , Papúa Nueva Guinea , Timor OrientalRESUMEN
Schistosoma mansoni is responsible for the neglected tropical disease schistosomiasis that affects 210 million people in 76 countries. Here we present analysis of the 363 megabase nuclear genome of the blood fluke. It encodes at least 11,809 genes, with an unusual intron size distribution, and new families of micro-exon genes that undergo frequent alternative splicing. As the first sequenced flatworm, and a representative of the Lophotrochozoa, it offers insights into early events in the evolution of the animals, including the development of a body pattern with bilateral symmetry, and the development of tissues into organs. Our analysis has been informed by the need to find new drug targets. The deficits in lipid metabolism that make schistosomes dependent on the host are revealed, and the identification of membrane receptors, ion channels and more than 300 proteases provide new insights into the biology of the life cycle and new targets. Bioinformatics approaches have identified metabolic chokepoints, and a chemogenomic screen has pinpointed schistosome proteins for which existing drugs may be active. The information generated provides an invaluable resource for the research community to develop much needed new control tools for the treatment and eradication of this important and neglected disease.
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Genoma de los Helmintos/genética , Schistosoma mansoni/genética , Animales , Evolución Biológica , Exones/genética , Genes de Helminto/genética , Interacciones Huésped-Parásitos/genética , Intrones/genética , Datos de Secuencia Molecular , Mapeo Físico de Cromosoma , Schistosoma mansoni/efectos de los fármacos , Schistosoma mansoni/embriología , Schistosoma mansoni/fisiología , Esquistosomiasis mansoni/tratamiento farmacológico , Esquistosomiasis mansoni/parasitologíaRESUMEN
Molecular diagnostics for crop diseases can enhance food security by enabling the rapid identification of threatening pathogens and providing critical information for the deployment of disease management strategies. Loop-mediated isothermal amplification (LAMP) is a PCR-based tool that allows the rapid, highly specific amplification of target DNA sequences at a single temperature and is thus ideal for field-level diagnosis of plant diseases. We developed primers highly specific for two globally important rice pathogens, Xanthomonas oryzae pv. oryzae, the causal agent of bacterial blight (BB) disease, and X. oryzae pv. oryzicola, the causal agent of bacterial leaf streak disease (BLS), for use in reliable, sensitive LAMP assays. In addition to pathovar distinction, two assays that differentiate X. oryzae pv. oryzae by African or Asian lineage were developed. Using these LAMP primer sets, the presence of each pathogen was detected from DNA and bacterial cells, as well as leaf and seed samples. Thresholds of detection for all assays were consistently 10(4) to 10(5) CFU ml(-1), while genomic DNA thresholds were between 1 pg and 10 fg. Use of the unique sequences combined with the LAMP assay provides a sensitive, accurate, rapid, simple, and inexpensive protocol to detect both BB and BLS pathogens.
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Técnicas de Amplificación de Ácido Nucleico/métodos , Oryza/microbiología , Enfermedades de las Plantas/microbiología , Xanthomonas/aislamiento & purificación , Cartilla de ADN/genética , ADN Bacteriano/genética , Sensibilidad y Especificidad , Xanthomonas/clasificación , Xanthomonas/genéticaRESUMEN
In this study, swine fecal specimens (n = 251) collected from nursing and weaned piglets raised under smallholder production systems were screened for the presence of kobuviruses by RT-PCR. Porcine kobuviruses were detected in 13.1 % (33/251) of the samples. We demonstrated that porcine kobuvirus infections exist in indigenous pigs in Kenya and Uganda and that the prevalence was higher in young piglets than older pigs: nursing piglets (15 %), post-weaning (3-month-old) pigs (17 %), 4-month-old pigs (10 %). Genetic analysis of the partial RNA-dependent RNA polymerase (RdRp) region (690 nt) revealed that kobuviruses circulating in East Africa are diverse, sharing nucleotide sequence identities ranging from 89.7 to 99.1 % and 88 to 92.3 % among them and with known porcine kobuviruses, respectively. The nucleotide sequence identities between our kobuvirus strains and those of human, bovine and canine kobuviruses were 69.4-70.7 %, 73.1-74.4 % and 67-70.7 %, respectively. Additionally, upon sequencing selected samples that showed consistent 720-bp RT-PCR bands while using the same primer set, we detected porcine astroviruses in our samples belonging to type 2 and type 3 mamastroviruses. To our knowledge, this study reports the first detection and molecular analysis of both porcine kobuviruses and astroviruses in an African region. Further studies are required to determine the role of these viruses in gastrointestinal infections of pigs in this region and to determine the genetic diversity of the circulating strains to develop accurate diagnostic tools and implement appropriate control strategies.
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Infecciones por Astroviridae/veterinaria , Astroviridae/aislamiento & purificación , Kobuvirus/aislamiento & purificación , Infecciones por Picornaviridae/veterinaria , Enfermedades de los Porcinos/virología , África Oriental/epidemiología , Animales , Infecciones por Astroviridae/epidemiología , Infecciones por Astroviridae/virología , Heces/virología , Variación Genética , Datos de Secuencia Molecular , Infecciones por Picornaviridae/epidemiología , Infecciones por Picornaviridae/virología , Prevalencia , ARN Viral/genética , ARN Polimerasa Dependiente del ARN/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Homología de Secuencia , Porcinos , Enfermedades de los Porcinos/epidemiologíaRESUMEN
Approximately 80% of the global cattle population is at risk of infestation and infection by ticks and tick-borne diseases (TTBDs). The economic losses from animal mortality, reduced production, vector control costs and animal treatment are very substantial, hence there is an urgent need to develop and deploy alternative vector control strategies. Breeding for host tick resistance has the potential for sustainable large-scale TTBD control especially in cattle. The gold standard method for phenotyping tick resistance in cattle is by counting ticks on the body but is very laborious and subjective. Better methods for phenotyping tick resistance more objectively, faster and at scale, are essential for selecting host genetic resistance to ticks. This study investigated the correlation between haematological cellular profiles and immunological responses (immunoglobulin E, IgE) and full body tick counts in herds of Bos indicus and Bos taurus following artificial tick challenge with Rhipicephalus decoloratus larvae. Fifty-four Friesian and Ayrshire (Bos taurus) and 52 East African Zebu (Bos indicus) calves were each infested with â¼2500 larvae. Near-replete adult female ticks (≥ 4.5 mm) were counted daily from Day 20-25. Blood and serum samples were obtained from each animal on Days 0 and 23 for cellular blood and IgE titre analysis, respectively. The indicine cattle were refractory to R. decoloratus infestation in comparison with the taurine breed (P < 0.0001). Repeated measurements of blood components pre-infestation revealed a significant (P < 0.05) association with tick count in IgE and red blood cells, haematocrit, and haemoglobin post-infestation. There was also a strong positive correlation between the tick counts and red blood cell numbers, haemoglobin, haematocrit, and IgE concentration (P < 0.0001) following tick challenge. The application of this approach to phenotype host resistance needs to be assessed using higher cattle numbers and with different tick species or genera.
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There is an alarming rate of human African trypanosomiasis recrudescence in many parts of sub-Saharan Africa. Yet, the disease has no successful chemotherapy. Trypanosoma lacks the enzymatic machinery for the de novo synthesis of purine nucleotides, and is critically dependent on salvage mechanisms. Inosine 5'-monophosphate dehydrogenase (IMPDH) is responsible for the rate-limiting step in guanine nucleotide metabolism. Here, we characterize recombinant Trypanosoma brucei IMPDH (TbIMPDH) to investigate the enzymatic differences between TbIMPDH and host IMPDH. Size-exclusion chromatography and analytical ultracentrifugation sedimentation velocity experiments reveal that TbIMPDH forms a heptamer, different from type 1 and 2 mammalian tetrameric IMPDHs. Kinetic analysis reveals calculated K m values of 30 and 1300 µ m for IMP and NAD, respectively. The obtained K m value of TbIMPDH for NAD is approximately 20-200-fold higher than that of mammalian enzymes and indicative of a different NAD binding mode between trypanosomal and mammalian IMPDHs. Inhibition studies show K i values of 3·2 µ m, 21 nM and 3·3 nM for ribavirin 5'-monophosphate, mycophenolic acid and mizoribine 5'-monophosphate, respectively. Our results show that TbIMPDH is different from its mammalian counterpart and thus may be a good target for further studies on anti-trypanosomal drugs.
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IMP Deshidrogenasa/aislamiento & purificación , Trypanosoma brucei brucei/enzimología , Secuencia de Aminoácidos , Animales , Sitios de Unión , Escherichia coli/genética , Escherichia coli/metabolismo , Humanos , Concentración de Iones de Hidrógeno , IMP Deshidrogenasa/antagonistas & inhibidores , IMP Deshidrogenasa/genética , IMP Deshidrogenasa/metabolismo , Inosina Monofosfato/química , Inosina Monofosfato/metabolismo , Cinética , Ácido Micofenólico/farmacología , NAD/metabolismo , Nucleótidos/farmacología , Multimerización de Proteína , Proteínas Recombinantes , Ribonucleósidos/farmacología , Alineación de Secuencia , Trypanosoma brucei brucei/genéticaRESUMEN
As it feeds upon cattle, Rhipicephalus (Boophilus) microplus is capable of transmitting a number of pathogenic organisms, including the apicomplexan hemoparasite Babesia bovis, a causative agent of bovine babesiosis. The R. microplus female gut transcriptome was studied for two cohorts: adult females feeding on a bovine host infected with B. bovis and adult females feeding on an uninfected bovine. RNA was purified and used to generate a subtracted cDNA library from B. bovis-infected female gut, and 4,077 expressed sequence tags (ESTs) were sequenced. Gene expression was also measured by a microarray designed from the publicly available R. microplus gene index: BmiGI Version 2. We compared gene expression in the tick gut from females feeding upon an uninfected bovine to gene expression in tick gut from females feeding upon a splenectomized bovine infected with B. bovis. Thirty-three ESTs represented on the microarray were expressed at a higher level in female gut samples from the ticks feeding upon a B. bovis-infected calf compared to expression levels in female gut samples from ticks feeding on an uninfected calf. Forty-three transcripts were expressed at a lower level in the ticks feeding upon B. bovis-infected female guts compared with expression in female gut samples from ticks feeding on the uninfected calf. These array data were used as initial characterization of gene expression associated with the infection of R. microplus by B. bovis.
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Babesia bovis/fisiología , Babesiosis/veterinaria , Enfermedades de los Bovinos/parasitología , Rhipicephalus/genética , Infestaciones por Garrapatas/veterinaria , Transcriptoma , Animales , Babesiosis/parasitología , Secuencia de Bases , Bovinos , Estudios de Cohortes , Biología Computacional , Etiquetas de Secuencia Expresada , Femenino , Tracto Gastrointestinal/parasitología , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Biblioteca de Genes , Datos de Secuencia Molecular , Análisis de Secuencia por Matrices de Oligonucleótidos , Distribución Aleatoria , Reacción en Cadena en Tiempo Real de la Polimerasa , Rhipicephalus/parasitología , Rhipicephalus/fisiología , Análisis de Secuencia de ADN , Infestaciones por Garrapatas/parasitologíaRESUMEN
Objective: To estimate gastroenteritis disease and its etiological agents in children under the age of 5 years living in South Africa. Methods: A mini literature review of pertinent articles published in ScienceDirect, PubMed, GoogleScholar, and Scopus was conducted using search terms: "Gastroenteritis in children," "Gastroenteritis in the world," Gastroenteritis in South Africa," "Prevalence of gastroenteritis," "Epidemiological surveillance of gastroenteritis in the world," and "Causes of gastroenteritis". Results: A total of 174 published articles were included in this mini review. In the last 20 years, the mortality rate resulting from diarrhea in children under the age of 5 years has declined and this is influenced by improved hygiene practices, awareness programs, an improved water and sanitation supply, and the availability of vaccines. More modern genomic amplification techniques were used to re-analyze stool specimens collected from children in eight low-resource settings in Asia, South America, and Africa reported improved sensitivity of pathogen detection to about 65%, that viruses were the main etiological agents in patients with diarrhea aged from 0 to 11 months but that Shigella, followed by sapovirus and enterotoxigenic Escherichia coli had a high incidence in children aged 12-24 months. In addition, co-infections were noted in nearly 10% of diarrhea cases, with rotavirus and Shigella being the main co-infecting agents together with adenovirus, enteropathogenic E. coli, Clostridium jejuni, or Clostridium coli. Conclusions: This mini review outlines the epidemiology and trends relating to parasitic, viral, and bacterial agents responsible for gastroenteritis in children in South Africa. An increase in sequence-independent diagnostic approaches will improve the identification of pathogens to resolve undiagnosed cases of gastroenteritis. Emerging state and national surveillance systems should focus on improving the identification of gastrointestinal pathogens in children and the development of further vaccines against gastrointestinal pathogens.
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Ticks and tick-borne diseases cause significant loss in livestock production with about 80% world's cattle at risk. The cost of chemical control is high and there is an ever-increasing tick resistance to chemical acaricides. Genetic selection as alternative long-term control strategy is constrained by laborious phenotyping using tick counts or scores. This study explored the use of host volatile semiochemicals that may be attractants or repellents to ticks as a phenotype for new tick resistance, with potential to be used as a proxy in selection programmes. Approximately 100 young cattle composed of Bos indicus and Bos taurus were artificially infested with 2,500 African blue tick, Rhipicephalus decoloratus larvae, with daily female tick (4.5 mm) counts taken from day 20 post-infestation. Volatile organic compounds were sampled from cattle before and after tick infestation by dynamic headspace collection, analysed by high-resolution gas chromatography (GC) and subjected to multivariate statistical analysis. Using 6-day repeated measure analysis, three pre-infestation GC peaks (BI938 - unknown, BI966 - 6-methyl-5-hepten-2-one and BI995 - hexyl acetate) and one post-infestation GC peak (AI933 - benzaldehyde / (E)-2-heptenal) were associated with tick resistance (P < 0.01 and P < 0.05 respectively). The high correlation coefficients (r = 0.66) between repeated records with all volatile compounds support the potential predictive value for volatile compounds in selective breeding programmes for tick resistance in cattle.
Asunto(s)
Enfermedades de los Bovinos , Rhipicephalus , Infestaciones por Garrapatas , Bovinos , Femenino , Animales , Infestaciones por Garrapatas/veterinaria , Rhipicephalus/genética , Fenotipo , Enfermedades de los Bovinos/genéticaRESUMEN
The control of tick-borne haemoparasites in cattle largely relies on the use of acaricide drugs against the tick vectors, with some vaccination also being used against selected pathogens. These interventions can be difficult in Africa, where accessibility and cost of vaccines can be issues, and the increasing resistance of tick vectors to the widely used acaricides is a complication to disease control. A potential complementary control strategy could be the exploitation of any natural host genetic resistance to the pathogens. However, there are currently very few estimates of the extent of host resistance to tick-borne haemoparasites, and a significant contributing factor to this knowledge gap is likely to be the difficulty of collecting appropriate samples and data in the smallholder systems that predominate livestock production in low- and middle-income countries, particularly at scale. In this study, we have estimated the heritability for the presence/absence of several important haemoparasite species (including Anaplasma marginale, Babesia bigemina, Babesia bovis, and Ehrlichia ruminantium), as well as for relevant traits such as body weight and body condition score (BCS), in 1,694 cattle from four African countries (Burkina Faso, Ghana, Nigeria, and Tanzania). Heritability estimates within countries were mostly not significant, ranging from 0.05 to 0.84 across traits and countries, with standard errors between 0.07 and 0.91. However, the weighted mean of heritability estimates was moderate and significant for body weight and BCS (0.40 and 0.49, respectively), with significant heritabilities also observed for the presence of A. marginale (0.16) and E. ruminantium (0.19). In a meta-analysis of genome-wide association studies (GWAS) for these traits, two peaks were identified as reaching the suggestive significance threshold (p < 1.91 × 10-7 and p < 1.89 × 10-7, respectively): one on chromosome 24 for BCS and one on chromosome 8 for the E. ruminantium infection status. These findings indicate that there is likely to be a genetic basis that contributes to pathogen presence/absence for tick-borne haemoparasite species, which could potentially be exploited to improve cattle resistance in Africa to the economically important diseases caused by these pathogens.
RESUMEN
The African livestock sector plays a key role in improving the livelihoods of people through the supply of food, improved nutrition and consequently health. However, its impact on the economy of the people and contribution to national GDP is highly variable and generally below its potential. This study was conducted to assess the current state of livestock phenomics and genetic evaluation methods being used across the continent, the main challenges, and to demonstrate the effects of various genetic models on the accuracy and rate of genetic gain that could be achieved. An online survey of livestock experts, academics, scientists, national focal points for animal genetic resources, policymakers, extension agents and animal breeding industry was conducted in 38 African countries. The results revealed 1) limited national livestock identification and data recording systems, 2) limited data on livestock production and health traits and genomic information, 3) mass selection was the common method used for genetic improvement with very limited application of genetic and genomic-based selection and evaluation, 4) limited human capacity, infrastructure, and funding for livestock genetic improvement programmes, as well as enabling animal breeding policies. A joint genetic evaluation of Holstein-Friesian using pooled data from Kenya and South Africa was piloted. The pilot analysis yielded higher accuracy of prediction of breeding values, pointing to possibility of higher genetic gains that could be achieved and demonstrating the potential power of multi-country evaluations: Kenya benefited on the 305-days milk yield and the age at first calving and South Africa on the age at first calving and the first calving interval. The findings from this study will help in developing harmonized protocols for animal identification, livestock data recording, and genetic evaluations (both national and across-countries) as well as in designing subsequent capacity building and training programmes for animal breeders and livestock farmers in Africa. National governments need to put in place enabling policies, the necessary infrastructure and funding for national and across country collaborations for a joint genetic evaluation which will revolutionize the livestock genetic improvement in Africa.