Isolation and Characterization of Coronavirus and Rotavirus Associated With Calves, in Central Part of Oromia, Ethiopia

Abstract Background: Coronavirus and Rotavirus are most commonly associated etiologies for calves’ diarrhea resulting in loss of productivity and economy of farmers. However, various facets of diarrheal disease caused by coronavirus and rotavirus in calves in Ethiopia are inadequately understood. A cross sectional study was conducted with the aim of isolation and molecular characterization of coronavirus and rotavirus from calves in central part of Oromia (Bishoftu, Sebata, Holeta and Addis Ababa), Ethiopia from November 2018 to May 2019. The four study areas were purposively selected and fecal samples were collected by simple random sampling for diagnosis of coronavirus and rotavirus infection by using antigen detection Enzyme linked immunosorbent assay (Ag-ELISA) kit. In addition, this study was carried out to have insight in prevalence and associated risk factors of coronavirus and rotavirus infection in calves. Result: During the study 83 diarrheic and 162 non-diarrheic fecal samples collected from calves less than 4 weeks of age were screened for coronavirus and rotavirus. Of the 83 diarrheic samples, 1 sample (1.2%) was positive for coronavirus antigen (Ag) and 6 samples (7.2%) were found to be positive for rotavirus antigen (Ag) by Ag-ELISA. All the non-diarrheic samples were negative for both coronavirus and rotavirus Ag. The overall prevalence of coronavirus and rotavirus infection in calves were estimated as 0.4% (1/245) and 2.45% (6/245) respectively. All samples (7) of ELISA test positive of both coronavirus and rotavirus were propagated in Madin Darby bovine kidney cells (MDBK). After 3 subsequent passages, progressive cytopathic effect (CPE) i.e. rounding, detachment as well as destruction of mono-layer cell of five sample (1 sample of coronavirus and 4 sample of rotavirus) (71.4%) were observed. At the molecular stage, reverse transcriptase polymerase chain reaction (RT-PCR) technique was used to determine the presence of coronavirus and rotavirus nucleic acid by using specific primers. The 5 samples that were coronavirus and rotavirus antigen positive by ELISA and develop CPE on cell culture were also positive on RT-PCR technique. Infection prevalence peaked have been obtained at 1st and 2nd weeks of age in male calves. Conclusion: Diarrheal disease caused by coronavirus and rotavirus has a great health problem in calves that interrupts production benefits with reduced weight gain and increased mortality, and its potential for zoonotic spread. So the present findings show coronavirus and rotavirus infection in calves in Ethiopia that needs to be addressed by practicing early colostrums feeding in newborn calves, using vaccine, or improving livestock management.

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Isolation Coronavirus and Rotavirus Associated With Calves, in Central Part of Oromia, Ethiopia

10.21203/rs.3.rs-39022/v2 ◽

2020 ◽

Author(s):

Umer Seid ◽

Fufa Dawo ◽

Munera Ahmednur ◽

Asamino Tesfaye

Keyword(s):

Diarrheal Disease ◽

Addis Ababa ◽

Simple Random Sampling ◽

Elisa Test ◽

Enzyme Linked Immunosorbent Assay ◽

Infection Prevalence ◽

Rt Pcr ◽

Cross Sectional ◽

Fecal Samples ◽

Rotavirus Infection

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Isolation and Characterization of Coronavirus and Rotavirus Associated with Calves in Central Part of Oromia, Ethiopia

Veterinary Medicine International ◽

10.1155/2020/8869970 ◽

2020 ◽

Vol 2020 ◽

pp. 1-10

Author(s):

Umer Seid ◽

Fufa Dawo ◽

Asamino Tesfaye ◽

Munera Ahmednur

Keyword(s):

Diarrheal Disease ◽

Addis Ababa ◽

Simple Random Sampling ◽

Enzyme Linked Immunosorbent Assay ◽

Rt Pcr ◽

Cross Sectional ◽

Rotavirus Infection ◽

Isolation And Characterization ◽

Faecal Samples

Background. Coronavirus and rotavirus are most commonly associated etiologies for calves’ diarrhoea, resulting in loss of productivity and economy of farmers. However, various facets of diarrheal disease caused by coronavirus and rotavirus in calves in Ethiopia are inadequately understood. A cross-sectional study was conducted with the aim of isolation and molecular characterization of coronavirus and rotavirus from calves in the central part of Oromia (Bishoftu, Sebata, Holeta, and Addis Ababa), Ethiopia, from November 2018 to May 2019. The four study areas were purposively selected and faecal samples were collected by simple random sampling for diagnosis of coronavirus and rotavirus infection by using the antigen detection enzyme-linked immunosorbent assay (Ag-ELISA) kit. In addition, this study was carried out to have insight in prevalence and associated risk factors of coronavirus and rotavirus infection in calves. Result. During the study, 83 diarrheic and 162 nondiarrheic faecal samples collected from calves less than 4 weeks of age were screened for coronavirus and rotavirus. Of the 83 diarrheic samples, 1 sample (1.2%) was positive for coronavirus antigen and 6 samples (7.2%) were found to be positive for rotavirus antigen by Ag-ELISA. All the nondiarrheic samples were negative for both coronavirus and rotavirus Ag. The overall prevalence of coronavirus and rotavirus infection in calves was estimated at 0.4% (1/245) and 2.45% (6/245), respectively. All samples (7) of ELISA test positive of both coronavirus and rotavirus were propagated in Madin-Darby bovine kidney (MDBK) cells. After 3 subsequent passages, progressive cytopathic effect (CPE), i.e., rounding, detachment, and the destruction of monolayer cell of five samples (1 sample of coronavirus and 4 samples of rotavirus) (71.4%) were observed. At the molecular stage, reverse transcriptase polymerase chain reaction (RT-PCR) technique was used to determine the presence of coronavirus and rotavirus nucleic acid by using specific primers. The 5 samples that were coronavirus and rotavirus antigen positive by ELISA and develop CPE on cell culture were also positive on RT-PCR technique. The prevalence of infection peaked at 1st and 2nd weeks of age in male calves. Conclusion. Diarrheal disease caused by coronavirus and rotavirus has a great health problem in calves that interrupts production benefits with reduced weight gain and increased mortality and its potential for zoonotic spread. So, the present findings show coronavirus and rotavirus infection in calves in Ethiopia that needs to be addressed by practising early colostrum feeding in newborn calves, using vaccine, or improving livestock management.

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Investigation of Rotavirus infection in sheep usingserological and molecular techniques

Indian Journal of Animal Research ◽

10.18805/ijar.11463 ◽

2016 ◽

Author(s):

Volkan Yilmaz. ◽

M.Ozkan Timurkan ◽

Nuvit Coskun ◽

Yakup Yildirim

Keyword(s):

Molecular Detection ◽

Molecular Techniques ◽

Enzyme Linked Immunosorbent Assay ◽

Small Scale ◽

Family Farms ◽

Rt Pcr ◽

Fecal Samples ◽

Rotavirus Infection ◽

Scale Family ◽

Polymerase Chain

In this study, serological and molecular research was conducted on the Rotavirus infection in domestic breeds of sheep at 2–3 years of age. The sheep included in the study were raised on small scale family units of less than 20 sheep per unit, in central Kars province and its districts (Susuz, Arpaçay, Kagizman and Selim) in the Northeast Anatolia region of Turkey. The blood and fecal samples were collected randomly from 450 sheep. They were analyzed for the presence of Rotavirus and the antibody against the virus using enzyme-linked immunosorbent assay (ELISA). The highest seropositive ratio (73.46%) was found in central Kars province. The seroprevalence of Rotavirus in sheep raised in the Kars region was determined to be 55.33%. Rotavirus was not detected in fecal samples with ELISA. Molecular detection of Rotavirus from fecal samples was done by reverse transcription polymerase chain reaction (RT-PCR) technique using specific generic primers for VP6 protein. Rotavirus could not be detected in RT-PCR. The data that were obtained showed that the infection spreads on small scale family farms. Based on this information, recommendations were made for controlling Rotavirus infection.

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Comparative study on lesions of reproductive disorders of cows and female dromedary camels slaughtered at Addis Ababa, Adama and Akaki abbatoirs with bacterial isolation and characterization

10.21203/rs.3.rs-35515/v3 ◽

2020 ◽

Author(s):

Aynalem Mandefro ◽

Tilaye Demissie Ayana ◽

Gemechu Chala Hunderra ◽

Tadesse Gidey Gebrezihar ◽

Bulto Giro Boru ◽

...

Keyword(s):

Escherichia Coli ◽

Addis Ababa ◽

Sampling Technique ◽

Normal Structure ◽

Bacterial Isolates ◽

Dromedary Camels ◽

Cross Sectional ◽

Tissue Samples ◽

Pathological Lesions ◽

Isolation And Characterization

Abstract Background: Reproduction is a basic prerequisite to efficient livestock production. Reproductive performance depends upon the normal structure and function of genital organs. Methods: A cross-sectional study was conducted from November 2016 to May 2017 to identify and compare the frequency of pathological lesions in the reproductive tract and to isolate bacteria associated to uterine lesions in female dromedary camels and cows slaughtered at Akaki camel slaughter house and Addis Ababa and Adama municipal abattoirs. Abattoirs were visited once per week for 28 weeks during which three to seven animals on average were slaughtered per day. A purposive sampling technique was employed to examine reproductive tracts of all slaughtered animals (280; 140 cows and 140 camels). Following gross inspection at abattoirs, tissue samples with lesion were collected for histopathological and bacteriological investigation. Result: Various pathological lesions with different degrees of severity were observed in 48 (34.2%) and 51 (36.4%) of dromedary camels and cows, respectively. Uterine lesions were the most prevalent 21.4% lesions observed in dromedary camels followed by ovarian lesions 7.14%; while in cows, ovarian lesions were the major prevalent 16.4% lesions followed by uterine lesion 14.2%. The result showed that there were 56 bacteria isolated from cows uterine lesion with Staphylococcus species 28.5%, Streptococci species 19.6%, Coynebacterium species 8.9%, Escherichia coli 26.78%, Salmonella species 10.7% and Klebsiella species 5.35% being the prominent isolates; while in camels, there were 45 bacteria isolated with Escherichia coli 35.5%, Staphylococcus species 26.6%, Streptococcus species 13.3%, Pseudomonas species 6.6 %, Proteus species 4.4%, Salmonella species 8.8% and Klebsiella species 4.4% being the most frequently isolated. The result showed that the major isolates were similar with slightly higher in cows. Histopathologically, endometrial glands degeneration, sloughing of epithelium, peri-glandular cuffing and infiltrations of inflammatory cell were some of characteristic changes observed in uterus. Conclusions: Pathological lesions in reproductive organs in female dromedary camels and cows showed great prevalence, with similarity in bacterial isolates between the two species. The role of each reproductive lesions and bacterial isolates incriminated as causes of reproductive failures in this livestock species needs further investigation.

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Isolation and Characterization of Mesenchymal Stem Cells from Chicken Liver

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2020.2225 ◽

2020 ◽

Vol 10 (1) ◽

pp. 8-16

Author(s):

Xibin Liu ◽

Shuang Zhang ◽

Weijun Guan ◽

Dong Zheng

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Protein Markers ◽

Multilineage Differentiation ◽

Rt Pcr ◽

Multipotent Stem Cells ◽

Isolation And Characterization ◽

Potential Applications ◽

Polymerase Chain

Hepatic mesenchymal stem cells (HMSCs) are multipotent stem cells that is a vital part of the regeneration of hepatocytes after injury. In this study, HMSCs were isolated in embryonic livers from of 12-day-old chick embryo using collagenase, and the primary HMSCs were sub-cultured to passage. The protein markers of HMSCs, namely CD71, CD29 and CD44, were tested with immunofluorescence and Reverse Transcription-Polymerase Chain Reaction (RT-PCR). The proliferation of HMSCs in different passages was detected using growth curve, which shown a typically sigmoidal. And then, the pluripotent of HMSCs was analyzed, the results showed that HMSCs could directly induce to differentiate into neural-like cells, adipocytes, and osteoblasts. Our data illustrated that the chick HMSCs have same characteristics to those obtained from other species. The capacity of these cells for multilineage differentiation shows promise for many potential applications.

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Distribution and genetic diversity of Enterovirus G (EV-G) on pig farms in Thailand

BMC Veterinary Research ◽

10.1186/s12917-021-02988-6 ◽

2021 ◽

Vol 17 (1) ◽

Author(s):

Taveesak Janetanakit ◽

Supassama Chaiyawong ◽

Kamonpan Charoenkul ◽

Ratanaporn Tangwangvivat ◽

Ekkapat Chamsai ◽

...

Keyword(s):

Genetic Diversity ◽

Genetic Characterization ◽

Rt Pcr ◽

Cross Sectional Survey ◽

High Genetic Diversity ◽

Cross Sectional ◽

Predominant Genotype ◽

Diarrhea Virus ◽

Pig Farms

Abstract Background Enterovirus G (EV-G) causes subclinical infections and is occasionally associated with diarrhea in pigs. In this study, we conducted a cross-sectional survey of EV-G in pigs from 73 pig farms in 20 provinces of Thailand from December 2014 to January 2018. Results Our results showed a high occurrence of EV-Gs which 71.6 % of fecal and intestinal samples (556/777) and 71.2 % of pig farms (52/73) were positive for EV-G by RT-PCR specific to the 5’UTR. EV-Gs could be detected in all age pig groups, and the percentage positivity was highest in the fattening group (89.7 %), followed by the nursery group (89.4 %). To characterize the viruses, 34 EV-G representatives were characterized by VP1 gene sequencing. Pairwise sequence comparison and phylogenetic analysis showed that Thai-EV-Gs belonged to the EV-G1, EV-G3, EV-G4, EV-G8, EV-G9 and EV-G10 genotypes, among which the EV-G3 was the predominant genotype in Thailand. Co-infection with different EV-G genotypes or with EV-Gs and porcine epidemic diarrhea virus (PEDV) or porcine deltacoronavirus (PDCoV) on the same pig farms was observed. Conclusions Our results confirmed that EV-G infection is endemic in Thailand, with a high genetic diversity of different genotypes. This study constitutes the first report of the genetic characterization of EV-GS in pigs in Thailand.

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Prevalence of rotavirus infection and its association with socio-demographic characteristics among under-five children admitted with acute diarrhoea in a tertiary care hospital of Bangladesh

BIRDEM Medical Journal ◽

10.3329/birdem.v11i3.55217 ◽

2021 ◽

Vol 11 (3) ◽

pp. 186-190

Author(s):

Farjana Binte Habib ◽

Mohammed Mirazur Rahman ◽

Rafaat Choudhury ◽

Tarun Kanti Paul ◽

Md Moynul Haque

Keyword(s):

Tertiary Care ◽

Acute Diarrhoea ◽

Watery Diarrhoea ◽

Enzyme Linked Immunosorbent Assay ◽

High Morbidity ◽

Care Hospital ◽

Cross Sectional ◽

Rotavirus Infection ◽

Under Five ◽

Under Five Children

Background: Rotavirus has been recognized as the most common cause of acute watery diarrhoea among under-five children worldwide. It is considered as a major cause of childhood morbidity and mortality particularly in developing countries like Bangladesh. Considering the high morbidity and significant mortality, this study was designed to evaluate the prevalence of rotaviral diarrhoea among children less than five years in Sylhet region of Bangladesh. Methods: This cross-sectional study was conducted in the Department of Microbiology, Sylhet MAG Osmani Medical College from 1st January to 31st December, 2018. Total 184 under-five children admitted in paediatrics department of the same institute with acute watery diarrhoea were enrolled in this study by convenient sampling. Stool samples were obtained and assayed for rotavirus antigen by sandwich type enzyme linked immunosorbent assay (ELISA). Results: Rotaviral antigen was found positive in 86 cases and disease prevalence was 46.74%. The rotavirus infection was found highest in age group of 7 to 12 months (50.56%) and in male (59.30%) children. Prevalence of infection was associated with age (p= 0.004), socio-economic condition (p=0.001) and exclusive breastfeeding of children (p=0.01). Conclusion: Rotavirus was a significant cause of acute watery diarrhoea and mainly affected children below 12 months age. BIRDEM Med J 2021; 11(3): 186-190

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Seroprevalence of brucellosis and molecular characterization of Brucella spp. from slaughtered cattle in Rwanda

10.1101/2021.12.07.471573 ◽

2021 ◽

Author(s):

Jean Bosco Ntivuguruzwa ◽

Francis Babaman Kolo ◽

Emil Ivan Mwikarago ◽

Henriette vanHeerden

Keyword(s):

Control Program ◽

Culture Method ◽

Cross Sectional Study ◽

Selective Medium ◽

Enzyme Linked Immunosorbent Assay ◽

Bovine Brucellosis ◽

Serological Tests ◽

Cross Sectional ◽

Standard Culture

AbstractBovine brucellosis is endemic in Rwanda, although, there is paucity of documented evidence about the disease in slaughtered cattle. A cross-sectional study was conducted in slaughtered cattle (n=300) to determine the seroprevalence of anti-Brucella antibodies using the Rose Bengal Test (RBT), and indirect enzyme-linked immunosorbent assay (i-ELISA). Corresponding tissues were cultured onto a modified Centro de Investigación y Tecnología Agroalimentaria (CITA) selective medium and analysed for Brucella spp. using the 16S-23S ribosomal interspacer region (ITS), AMOS, and Bruce-ladder PCR assays. The RBT seroprevalence was 20.7% (62/300), and 2.9% (8/300) with i-ELISA and 2.9% (8/300) using both tests in parallel. Brucella specific 16S-23S ribosomal DNA interspace region (ITS) PCR detected Brucella DNA in 5.6% (17/300; Brucella culture prevalence). AMOS-PCR assay identified mixed B. abortus and B. melitensis (n=3), B. abortus (n=3) and B. melitensis (n=5) while Bruce-ladder PCR also identified B. abortus (n=5) and B. melitensis (n=6). The gold standard culture method combined with PCR confirmation identified 5.6% Brucella cultures which is higher than the more sensitive seroprevalence of 2.9%. This emphasizes the need to validate the serological tests in Rwanda. The mixed infection caused by B. abortus and B. melitensis in slaughtered cattle indicates cross-infection and poses a risk of exposure potential to abattoir workers. It is essential to urgently strengthen the national bovine brucellosis control program through vaccination as well as test- and-slaughter.

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Sero-prevalence of anti-SARS-CoV-2 Antibodies in Addis Ababa, Ethiopia

10.1101/2020.10.13.337287 ◽

2020 ◽

Author(s):

Berhanu Nega Alemu ◽

Adamu Addissie ◽

Gemechis Mamo ◽

Negussie Deyessa ◽

Tamrat Abebe ◽

...

Keyword(s):

Serological Test ◽

Addis Ababa ◽

Rapid Test ◽

Cross Sectional Study ◽

Rt Pcr ◽

Serological Tests ◽

Cross Sectional ◽

History Of ◽

Community Transmission ◽

Pcr Test

AbstractBackgroundAnti-SARS-CoV-2 antibody tests are being increasingly used for sero-epidemiological purposes to provide better understanding of the extent of the infection in the community, and monitoring the progression of the COVID-19 epidemic. We conducted sero-prevalence study to estimate prior infection with with SARS-CoV-2 in Addis Ababa.MethodsA cross-sectional study was done from April 23 to 28, 2020 among 301 randomly selected residents of Addis Ababa; with no known history of contact with confirmed COVID-19 person. Interviews on socio demographic and behavioural risk factor followed by serological tests were performed for SARS-CoV-2 IgM, and IgG antibodies, using COVID-19 IgG/IgM Rapid Test Cassette. The test has sensitivity of 87·9% and specificity of 100% for lgM; and a sensitivity of 97·2% and specificity of 100% for IgG. RT-PCR test was also done on combined nasopharyngeal and oropharengeal swabs as an important public health consideration.FindingsThe unadjusted antibody-based crude SARS-CoV-2 prevalence was 7·6% and the adjusted true SARS-CoV-2 prevalence was estimated at 8·8% (95% CI 5·5%-11·6%) for the study population. Higher sero-prevalence were observed for males (9.0%), age below 50 years (8.2%), students and unemployed (15.6%), those with primary education (12.1%), smokers (7.8%), alcohol consumers (8.6%), chatt-chewers (13.6%) and shish smokers (18.8%). Seroprevalence was not significantly associated neither with socio-demographic not behavioral characteristics. According to the findings, possibly more individuals had been infected in Addis Ababa than what was being detected and reported by RT-PCR test suggestive of community transmission. The use of serological test for epidemiological estimation of the extent of SARS-CoV-2 epidemic gives a more precise estimate of magnitude which would be used for further monitoring and surveillance of the magnitude of the SARS CoV-2 infection.

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Pestivirus Infections in Semi-Domesticated Eurasian Tundra Reindeer (Rangifer tarandus tarandus): A Retrospective Cross-Sectional Serological Study in Finnmark County, Norway

Viruses ◽

10.3390/v12010029 ◽

2019 ◽

Vol 12 (1) ◽

pp. 29 ◽

Cited By ~ 1

Author(s):

Carlos G. das Neves ◽

Jonas Johansson Wensman ◽

Ingebjørg Helena Nymo ◽

Eystein Skjerve ◽

Stefan Alenius ◽

...

Keyword(s):

Rangifer Tarandus ◽

Enzyme Linked Immunosorbent Assay ◽

Border Disease Virus ◽

Serum Samples ◽

Cross Sectional ◽

Diarrhea Virus ◽

Border Disease ◽

Serological Studies ◽

Viral Diarrhea Virus

Members of the Pestivirus genus (family Flaviviridae) cause severe and economically important diseases in livestock. Serological studies have revealed the presence of pestiviruses in different cervid species, including wild and semi-domesticated Eurasian tundra reindeer. In this retrospective study, serum samples collected between 2006 and 2008 from 3339 semi-domesticated Eurasian reindeer from Finnmark County, Norway, were tested for anti-pestivirus antibodies using an enzyme linked immunosorbent assay (ELISA) and a subset of these by virus neutralization test (VNT). A seroprevalence of 12.5% was found, varying from 0% to 45% among different herding districts, and 20% in western Finnmark, as compared to 1.7% in eastern Finnmark. Seroprevalence increased with age. Pestivirus-specific RNA was not detected in any of the 225 serum samples tested by real-time RT-PCR. Based on VNT results, using a panel of one bovine viral diarrhea virus (BVDV) strain and two border disease virus (BDV) strains, the virus is most likely a reindeer-specific pestivirus closely related to BDV. A characterization of the causative virus and its pathogenic impact on reindeer populations, as well as its potential to infect other domestic and wild ruminants, should be further investigated.

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