Detection And Characterization Of Chicken Anemia Virus Isolated From Commercial Broiler Breeder Farms In Malaysia

Chicken anemia virus (CAV) is the causative agent of chicken infectious anemia (CIA). It is an economically important pathogen with a world-wide distribution. Study on the type of CAV isolates present and their genetic diversity, transmission to their progeny and level of protection afforded in the breeder farms is lacking in Malaysia. Hence, the present study was aimed to detect CAV from commercial broiler breeder farms using molecular, serological and immunohistochemical methods and characterize CAV positive samples based on sequence and phylogenetic analysis of partial VP1 gene. In the present study CAV DNA was detected in all 60 commercial broiler breeder hens obtained from 12 farms in three states of Malaysia. Results from ELISA also showed that 96.15% of blood samples collected from the same farms were positive for antibody against CAV supporting the finding from the nested PCR assay. Both of these findings indicate that CAV is widespread in commercial broiler breeder hens at least in the three states of Malaysia. Testing pooled embryonic tissue samples consisting of thymus, bursa of Fabricius and spleen together with egg shell membrane (ESM) showed positive embryos for CAV DNA in the range of 40% to 100% for different commercial broiler breeder farms despite the presence of neutralizing antibodies in majority of the hens (96.15%) tested for CAV antibodies. This shows high level of occurrence of vertical transmission of viral DNA to the progeny. The CAV antigen was also detected in the lymphocytes within the cortex of the thymus and in the hemocytoblasts of the bone marrow by indirect immunoperoxidase staining in some birds. The analysis of 165 amino acid portion of the VP1 protein of 12 isolates from commercial broiler breeder farms revealed unique amino acid residues proline (P) at amino acid position 22 and glutamine (G) at amino acid position 48 in isolates NF4A and PYT4, respectively. Generally, isolates from the commercial broiler breeder farms can be grouped into two based on their amino acid profile at positions 75, 97, 139 and 144. Seven of the isolates (NF4A, PPW4, P24A, P12B, M3B5, MF3C and MF1A) from the commercial broiler breeder farms had 75-I, 97-L, 139-Q and 144-Q and clustered together in cluster IIIa of the deduced amino acid phylogenetic tree whilst the remaining five isolates (M1B1, NF1D, NF2C, NF3A and PYT4) had similar 75-V, 97-M, 139-K and 144-E profile and found in cluster I and II of the deduced amino acid phylogenetic tree. When compared with previously published local field isolates, six isolates from the commercial broiler breeder farms (MF1A, MF3C, M3B5, NF4A, P12B and P24A) were found to have maximum homology with SMSC-1 isolate, four isolates (M1B1, NF3A, PYT4 and PPW4) were found to have maximum homology with BL-5 isolate and the remaining two (NF1D and NF2C) have similar maximum homology both with isolates 3-1 and BL-5. The sequence and phylogenetic analysis further indicated high similarity of current isolates from the commercial broiler breeder farms with isolates in this part of the globe while still having limited variation with isolates from different geographical places. The importance of unique amino acid substitutions observed in this study requires further research in order to identify the detail characteristics of the isolates

Detection and characterization of chicken anemia virus from commercial broiler breeder chickens

<p>Abstract</p> <p>Background</p> <p>Chicken anemia virus (CAV) is the causative agent of chicken infectious anemia (CIA). Study on the type of CAV isolates present and their genetic diversity, transmission to their progeny and level of protection afforded in the breeder farms is lacking in Malaysia. Hence, the present study was aimed to detect CAV from commercial broiler breeder farms and characterize CAV positive samples based on sequence and phylogenetic analysis of partial VP1 gene.</p> <p>Results</p> <p>A total of 12 CAV isolates from different commercial broiler breeder farms were isolated and characterized. Detection of CAV positive embryos by the PCR assay in the range of 40 to 100% for different farms indicated high level of occurrence of vertical transmission of viral DNA to the progeny. CAV antigen was detected in the thymus and in the bone marrow but not in spleen, liver, duodenum, ovary and oviduct by indirect immunoperoxidase staining. The 12 CAV isolates were characterized based on partial sequences of VP1 gene. Six isolates (MF1A, MF3C, M3B5, NF4A, P12B and P24A) were found to have maximum homology with previously characterized Malaysian isolate SMSC-1, four isolates (M1B1, NF3A, PYT4 and PPW4) with isolate BL-5 and the remaining two (NF1D and NF2C) have maximum homology both with isolates 3-1 and BL-5. Meanwhile, seven of the isolates with amino acid profile of 75-I, 97-L, 139-Q and 144-Q were clustered together in cluster I together with other isolates from different geographical places. The remaining five isolates with amino acid profile of 75-V, 97-M, 139-K and 144-E were grouped under cluster II. All the CAV isolates demonstrated omega values (K_a/K_s) of less than one (the values ranging from 0.07 to 0.5) suggesting the occurrence of purifying (negative) selection in all the studied isolates.</p> <p>Conclusion</p> <p>The present study showed that CAV is widespread in the studied commercial broiler breeder farms. The result also indicated the occurrence of genetic variability in local CAV isolates that can be divided at least into two groups based on characteristic amino acid substitutions at positions 75, 97, 139 and 144 of the VP1 protein.</p

Household Readiness to Care for Mild and Asymptomatic COVID-19 Cases at Home, Southwest Ethiopia: A Community-based Cross-Sectional Study

BACKGROUND: Corona virus disease (COVID-19) continued with its notorious effects overwhelming health institutions. Thus, home-based identification and care for asymptomatic and mild cases of COVID-19 has been recommended. Therefore, the objective of this study was to assess the level of household readiness for caring asymptomatic and mild cases of COVID-19 at home.&nbsp; METHODS: A community-based cross-sectional study was conducted from March-June 2021 on randomly selected 778 households. Data entry and analysis were carried out using EpiData and SPSS version 25, respectively. Multivariable logistic regression was modeled to identify independent predictors of community readiness.&nbsp; RESULTS: Overall readiness of the community was very low (43.8%). Factors positively affecting household readiness were male household heads (AOR = 1.6; 95%CI: 1.05, 2.45), primary (AOR=2.0; CI:.62, 1.59) and higher (AOR = 1.90; 95%CI: 1.04, 3.45) educational level of the respondents, number of rooms within household (AOR = 1.22; CI: 1.03, 1.46), having additionally house (AOR = 2.61; CI: 1.35, 5.03), availability of single use eating utensils (AOR = 2.76; 95%CI: 1.66, 4.56), availability of community water supply (AOR = 8.21; 95% CI: 5.02, 13.43), and community participation and engagement (AOR = 2.81; 95% CI: 1.93, 4.08) in accessing transport, water and sanitation. CONCLUSIONS: The community was less prepared in terms of housing, infection prevention, water and sanitation. Considering alternative options including universal coverage of vaccine is important; designed behavioral change communications can enhance community participation and engagement in improving access to transport, water and sanitation to reduce risk of infections

Omega-3 polyunsaturated fatty acids enrichment alters performance and immune response in infectious bursal disease challenged broilers.

Background Infectious bursal disease (IBD) results in economic loss due to mortality, reduction in production efficiency and increasing the usage of antibiotics. This study was carried out to investigate the modulatory roles of dietary n-3 polyunsaturated fatty acids (PUFA) enrichment in immune response and performance of IBD challenged broiler chickens. Methods A total of 300 day old male broiler chicks were assigned to four dietary n-3 PUFA ascending levels as the treatment groups (T1: 0.5; T2: 8.0; T3: 11.5; T4: 16.5) using combinations of tuna oil and sunflower oil. All diets were isocaloric and isonitrogenous. On day 28, all birds were challenged with IBD virus. Antibody titer, cytokine production, bursa lesion pre and post-challenge and lymphoid organ weight were recorded. Results On d 42 the highest body weight was observed in the T2 and T3 and the lowest in T4 chickens. Feed conversion ratio of the T2 broilers was significantly better than the other groups. Although productive parameters were not responded to the dietary n-3 PUFA in a dose-dependent manner, spleen weight, IBD and Newcastle disease antibody titers and IL-2 and IFN-γ concentrations were constantly elevated by n-3 PUFA enrichment. Conclusions Dietary n-3 PUFA enrichment may improve the immune response and IBD resistance, but the optimum performance does not coincide with the optimum immune response. It seems that dietary n-3 PUFA modulates the broiler chicken performance and immune response in a dose-dependent manner. Thus, a moderate level of dietary n-3 PUFA enrichment may help to put together the efficiency of performance and relative immune response enhancement in broiler chickens

Global, regional, and national burden of chronic kidney disease, 1990–2017 : a systematic analysis for the Global Burden of Disease Study 2017

Background Health system planning requires careful assessment of chronic kidney disease (CKD) epidemiology, but data for morbidity and mortality of this disease are scarce or non-existent in many countries. We estimated the global, regional, and national burden of CKD, as well as the burden of cardiovascular disease and gout attributable to impaired kidney function, for the Global Burden of Diseases, Injuries, and Risk Factors Study 2017. We use the term CKD to refer to the morbidity and mortality that can be directly attributed to all stages of CKD, and we use the term impaired kidney function to refer to the additional risk of CKD from cardiovascular disease and gout. Methods The main data sources we used were published literature, vital registration systems, end-stage kidney disease registries, and household surveys. Estimates of CKD burden were produced using a Cause of Death Ensemble model and a Bayesian meta-regression analytical tool, and included incidence, prevalence, years lived with disability, mortality, years of life lost, and disability-adjusted life-years (DALYs). A comparative risk assessment approach was used to estimate the proportion of cardiovascular diseases and gout burden attributable to impaired kidney function. Findings Globally, in 2017, 1·2 million (95% uncertainty interval [UI] 1·2 to 1·3) people died from CKD. The global all-age mortality rate from CKD increased 41·5% (95% UI 35·2 to 46·5) between 1990 and 2017, although there was no significant change in the age-standardised mortality rate (2·8%, −1·5 to 6·3). In 2017, 697·5 million (95% UI 649·2 to 752·0) cases of all-stage CKD were recorded, for a global prevalence of 9·1% (8·5 to 9·8). The global all-age prevalence of CKD increased 29·3% (95% UI 26·4 to 32·6) since 1990, whereas the age-standardised prevalence remained stable (1·2%, −1·1 to 3·5). CKD resulted in 35·8 million (95% UI 33·7 to 38·0) DALYs in 2017, with diabetic nephropathy accounting for almost a third of DALYs. Most of the burden of CKD was concentrated in the three lowest quintiles of Socio-demographic Index (SDI). In several regions, particularly Oceania, sub-Saharan Africa, and Latin America, the burden of CKD was much higher than expected for the level of development, whereas the disease burden in western, eastern, and central sub-Saharan Africa, east Asia, south Asia, central and eastern Europe, Australasia, and western Europe was lower than expected. 1·4 million (95% UI 1·2 to 1·6) cardiovascular disease-related deaths and 25·3 million (22·2 to 28·9) cardiovascular disease DALYs were attributable to impaired kidney function. Interpretation Kidney disease has a major effect on global health, both as a direct cause of global morbidity and mortality and as an important risk factor for cardiovascular disease. CKD is largely preventable and treatable and deserves greater attention in global health policy decision making, particularly in locations with low and middle SDI

Global, regional, and national cancer incidence, mortality, years of life lost, years lived with disability, and disability-Adjusted life-years for 29 cancer groups, 1990 to 2017 : A systematic analysis for the global burden of disease study

Importance: Cancer and other noncommunicable diseases (NCDs) are now widely recognized as a threat to global development. The latest United Nations high-level meeting on NCDs reaffirmed this observation and also highlighted the slow progress in meeting the 2011 Political Declaration on the Prevention and Control of Noncommunicable Diseases and the third Sustainable Development Goal. Lack of situational analyses, priority setting, and budgeting have been identified as major obstacles in achieving these goals. All of these have in common that they require information on the local cancer epidemiology. The Global Burden of Disease (GBD) study is uniquely poised to provide these crucial data. Objective: To describe cancer burden for 29 cancer groups in 195 countries from 1990 through 2017 to provide data needed for cancer control planning. Evidence Review: We used the GBD study estimation methods to describe cancer incidence, mortality, years lived with disability, years of life lost, and disability-Adjusted life-years (DALYs). Results are presented at the national level as well as by Socio-demographic Index (SDI), a composite indicator of income, educational attainment, and total fertility rate. We also analyzed the influence of the epidemiological vs the demographic transition on cancer incidence. Findings: In 2017, there were 24.5 million incident cancer cases worldwide (16.8 million without nonmelanoma skin cancer [NMSC]) and 9.6 million cancer deaths. The majority of cancer DALYs came from years of life lost (97%), and only 3% came from years lived with disability. The odds of developing cancer were the lowest in the low SDI quintile (1 in 7) and the highest in the high SDI quintile (1 in 2) for both sexes. In 2017, the most common incident cancers in men were NMSC (4.3 million incident cases); tracheal, bronchus, and lung (TBL) cancer (1.5 million incident cases); and prostate cancer (1.3 million incident cases). The most common causes of cancer deaths and DALYs for men were TBL cancer (1.3 million deaths and 28.4 million DALYs), liver cancer (572000 deaths and 15.2 million DALYs), and stomach cancer (542000 deaths and 12.2 million DALYs). For women in 2017, the most common incident cancers were NMSC (3.3 million incident cases), breast cancer (1.9 million incident cases), and colorectal cancer (819000 incident cases). The leading causes of cancer deaths and DALYs for women were breast cancer (601000 deaths and 17.4 million DALYs), TBL cancer (596000 deaths and 12.6 million DALYs), and colorectal cancer (414000 deaths and 8.3 million DALYs). Conclusions and Relevance: The national epidemiological profiles of cancer burden in the GBD study show large heterogeneities, which are a reflection of different exposures to risk factors, economic settings, lifestyles, and access to care and screening. The GBD study can be used by policy makers and other stakeholders to develop and improve national and local cancer control in order to achieve the global targets and improve equity in cancer care. © 2019 American Medical Association. All rights reserved.Peer reviewe

Molekularbiologischer Nachweis und Charakterisierung von durch Zecken übertragenen Pathogenen bei Rindern im Südwesten Äthiopiens

Tick-borne diseases (TBDs) affect 80% of the world’s cattle population, hampering livestock production throughout the world. Bovine babesiosis, anaplasmosis, heartwater and theileriosis have all been reported to occur in Ethiopia. However, little is known about the extent and prevalence of these diseases in the local cattle population. The principal aim of this thesis was to estimate the species composition and prevalence of bovine tickborne pathogens (TBPs) in Southwestern Ethiopia. The first paper of this cumulative thesis deals with the evaluation of six DNA extraction methods from blood spotted on Flinders Technology Associates (FTA) cards, to determine the optical protocol for the subsequent molecular detection of TBPs by PCR and reverse line blot (RLB) hybridization. Ten-fold serial dilutions of bovine blood infected with Babesia bovis, Theileria mutans, Anaplasma marginale or Ehrlichia ruminantium were made by dilution with uninfected blood and spotted on FTA cards. Subsequently, DNA was extracted from FTA cards using six different DNA extraction protocols. Additionally, DNA was also isolated from whole blood dilutions using a commercial kit. The findings from this study showed that (i) among the methods compared, the highest detection limit was observed when DNA was extracted from FTA cards using FTA purification reagent combined with elution with 5% Chelex® 100 resin. Secondly, the detection limit was improved when more discs were used as starting material for the DNA extraction, whereby the use of sixteen 3 mm discs proved to be most practical, (iii) comparisons of the detection limits of RLB and PCR for the four TBPs of cattle showed that the sensitivity of DNA extraction from FTA card vary between different TBPs and the sensitivity of the RLB was in nearly all instances 10x higher compared to gel electrophoresis. The second study aimed to determine the prevalence and species composition of bovine TBPs of veterinary significance in local cattle populations of Illubabor zone in Southwestern Ethiopia. The best method for the extraction of DNA from blood spotted on FTA cards as described above and a combination of PCR and a Reverse Line Blot (RLB) hybridization assay were employed for the detection of a broad-spectrum of TBPs (i.e. Babesia, Theileria, Anaplasma and Ehrlichia spp.) in blood samples collected from 392 cattle. The findings showed a high prevalence of several bovine TBPs in Southwest Ethiopia, some which are of great veterinary importance. Individual prevalences of these pathogens included Theileria mutans (66.1%), Theileria orientalis (51.8%), Anaplasma sp. Omatjenne (25.5%), Anaplasma marginale (14.5%), Babesia bigemina (14.0%) and Theileria velifera (13.0%) and minor occurrences of Ehrlichia ruminantium (0.5%) and Ehrlichia minasensis (0.26%). Ehrlichia minasensis was until now only reported from North and South America. In addition, three novel Anaplasma genotypes were detected in bovine blood samples. A phylogenetic analysis revealed that they most likely represent three, but at least two, new species. A total of 227 cattle (57.9%) were found to be coinfected with two or more TBPs simultaneously and 86 different species combinations were observed. The high frequency of co-infections observed in this study suggests that clinical manifestations might be complex and complicate their diagnosis. The data presented in this thesis provides a more informed picture of the epidemiology of infection of cattle with TBPs and contributes towards integrated and strategic control of ticks and TBDs in Ethiopia.Zecken-übertragene Krankheiten (TBDs), welche 80 % der weltweiten Rinderpopulation betreffen, haben einen negativen Einfluss auf die Nutzviehwirtschaft auf der ganzen Welt. Das Vorkommen der Babesiose, Anaplasmose, Herzwasserkrankheit und Theileriose bei Rindern in Äthiopien wurde beschrieben. Allerdings ist wenig bekannt über die Bedeutung und die Prävalenz dieser Krankheiten in den lokalen Rinderpopulationen Äthiopiens. Ziel dieser Dissertation war es daher, die Spezieszusammensetzung und die Prävalenz der von Zecken übertragenen Pathogene (TBP) in Rindern im Südwesten von Äthiopien zu bestimmen. Die Dissertation gliedert sich in zwei Studien. Die erste Studie beschäftigt sich mit der Evaluierung von sechs DNS-Extraktionsmethoden. Eine Zehner-Verdünnungsreihe von Rinderblut, das mit Babesia bovis, Theileria mutans, Anaplasma marginale oder Ehrlichia ruminantium infiziert war, wurde durch Verdünnung mit nicht infiziertem Blut hergestellt und auf FTA-Karten („Flinders-Technology-Associates“ cards) übertragen. Anschließend wurde die DNS unter Verwendung von sechs verschiedenen DNS-Extraktionsmethoden aus den FTA-Karten extrahiert. Zusätzlich wurde vergleichend DNS aus den Proben mit einem handelsüblichen Extraktions-Kit isoliert. Ziel war es, das optimale Protokoll für eine sich anschließende molekulare Erfassung der von Zecken übertragenen Pathogene durch PCR und „Reverse-Line-Blot-Hybridisierung“ (RLB) zu bestimmen. Die Ergebnisse dieser Studie zeigten, dass (i) unter den verglichenen Methoden die höchste Nachweisempfindlichkeit für die Erreger erreicht wurde, wenn die DNS aus FTAKarten unter Verwendung der FTA-Reinigungslösung, kombiniert mit der Elution mit 5% Chelex® 100-Harz, extrahiert wurde. Zweitens wurde die Nachweisgrenze verbessert, wenn mehrere Stanzproben von den FTA-Karten der gleichen Probe als Ausgangsmaterial für die DNS-Extraktion verwendet wurden, wobei die Verwendung von sechzehn 3 mm-Stanzproben sich als am praktischsten erwies. (iii) Vergleiche der Nachweisgrenzen von RLB und PCR für die durch Zecken übertragenen Pathogene für Rinder zeigten, dass die Empfindlichkeit der DNS-Extraktion aus den FTA-Karten zwischen verschiedenen TBPs variierte und die Empfindlichkeit der RLB-Hybridisierung in fast allen Fällen 10x höher war als im Vergleich zur Gelelektrophorese. In der zweiten Studie sollte die Prävalenz und Spezieszusammensetzung der TBPs in den lokalen Rinderpopulationen in der Region von Illubator, im Südwesten von Äthiopien, bestimmt werden. Die Extraktionsmethode mit der höchsten Nachweisempfindlichkeit (FTA-Reinigungslösung kombiniert mit 5% Chelex® 100-Harz) wurde in Kombination mit der PCR und einem RLB-Hybridisierungsassay für die Detektion eines breiten Spektrums von TBPs angewandt. Insgesamt wurden Blutproben von 392 Rinder auf die Erreger aus vier TBP-Gattungen untersucht (Babesia spp., Theileria spp., Anaplasma spp. und Ehrlichia spp.). Die Ergebnisse zeigen eine hohe Prävalenz mehrerer Rinder-TBPs in Südwest-Äthiopien, von denen einige eine große veterinärmedizinische Bedeutung haben. Die Prävalenz dieser Pathogene betrug für Theileria mutans (66,1%), Theileria orientalis (51,8%), Anaplasma sp. Omatjenne (25,5%), Anaplasma marginale (14,5%), Babesia bigemina (14,0%), Theileria velifera (13,0%), Ehrlichia ruminantium (0,5%) und Ehrlichia minasensis (0,26%). Ehrlichia minasensis war zuvor ausschließlich in Rinderproben aus Nord- und Südamerika nachgewiese worden. Zusätzlich wurden drei neue Anaplasma-Genotypen in Rinderblutproben identifiziert. Eine phylogenetische Analyse zeigte ferner, dass diese höchstwahrscheinlich drei, zumindest aber zwei, neue Arten darstellen. Insgesamt waren 227 Rinder (57,9%) mit zwei oder mehreren TBPs gleichzeitig infiziert. Dabei wurden 86 verschiedene Spezieskombinationen beobachtet. Die in dieser Studie beschriebene Häufigkeit von Ko-Infektionen deutet darauf hin, dass klinische Manifestationen komplex sein können und daher eine Diagnose verkomplizieren können. Die in dieser Arbeit präsentierten Daten ergeben einen Überblick über das Artenspektrum und die Prävalenz der von Zecken übertragenen Pathogene bei Rindern im Südwesten von Äthiopien. Die epidemiologischen Daten liefern eine wichtige Voraussetzung für eine strategische und nachhaltige Kontrolle der Zecken und der durch sie im Untersuchungsgebiet übertragenen Erreger

Adaptation of Proteasomes and Lysosomes to Cellular Environments

Protein degradation is important for proper cellular physiology as it removes malfunctioning proteins or can provide a source for energy. Proteasomes and lysosomes, through the regulatory particles or adaptor proteins, respectively, recognize proteins destined for degradation. These systems have developed mechanisms to allow adaptation to the everchanging environment of the cell. While the complex recognition of proteins to be degraded is somewhat understood, the mechanisms that help switch the proteasomal regulatory particles or lysosomal adaptor proteins to adjust to the changing landscape of degrons, during infections or inflammation, still need extensive exploration. Therefore, this review is focused on describing the protein degradation systems and the possible sensors that may trigger the rapid adaptation of the protein degradation machinery

Aquatic Weed for Concrete Sustainability

Ordinary Portland cement (OPC) is the primary binder of concrete, accounting for approximately 5% to 7% of greenhouse gas (GHG) and carbon dioxide (CO2) emissions with an annual production rate of more than 4 billion tons. It is critical to reduce the carbon footprint of concrete without sacrificing its performance. To this end, this study focuses on the use of water hyacinth ash (WHA) as a pozzolanic binder in the production of concrete as a partial replacement for cement. Four mixes are designed to achieve C-25-grade concrete with varying proportions of cement replacement with WHA of 0%, 5%, 10%, and 15% of the cement weight. Extensive experiments are performed to examine the workability, strength, durability, and microstructure of concrete specimens. The test results confirm that incorporating WHA in concrete improved its workability, strength, and durability. The optimal results are obtained at the maximum OPC replacement level, with 10% WHA. The use of WHA as a partial replacement for cement greatly reduces the energy required for cement production and preserves natural resources. More research is needed to use WHA on a large scale to achieve greater sustainability in the concrete industry