scholarly journals Detection of Clinical and Subclinical Lumpy Skin Disease Using Ear Notch Testing and Skin Biopsies

2021 ◽  
Vol 9 (10) ◽  
pp. 2171
Author(s):  
Laetitia Aerts ◽  
Andy Haegeman ◽  
Ilse De Leeuw ◽  
Wannes Philips ◽  
Willem Van Campe ◽  
...  
Keyword(s):  
Skin Disease ◽  
Normal Skin ◽  
Viral Dna ◽  
Lumpy Skin Disease ◽  
Blood Samples ◽  
Clinical Surveillance ◽  
Fit For Purpose ◽  
Skin Biopsies ◽  
Unaffected Skin ◽  
Support Research

Lumpy skin disease (LSD) diagnosis is primarily based on clinical surveillance complemented by PCR of lesion crusts or nodule biopsies. Since LSD can be subclinical, the sensitivity of clinical surveillance could be lower than expected. Furthermore, real-time PCR for the detection of LSD viral DNA in blood samples from subclinical animals is only intermittently positive. Therefore, this study aimed to investigate an acceptable, easily applicable and more sensitive testing method for the detection of clinical and subclinical LSD. An animal experiment was conducted to investigate ear notches and biopsies from unaffected skin taken from the neck and dorsal back as alternatives to blood samples. It was concluded that for early LSD confirmation, normal skin biopsies and ear notches are less fit for purpose, as LSDV DNA is only detectable in these samples several days after it is detectable in blood samples. On the other hand, blood samples are less advisable for the detection of subclinical animals, while ear notches and biopsies were positive for LSD viral DNA in all subclinically infected animals by 16 days post infection. In conclusion, ear notches could be used for surveillance to detect subclinical animals after removing the clinical animals from a herd, to regain trade by substantiating the freedom of disease or to support research on LSDV transmission from subclinical animals.

scholarly journals The detection of lumpy skin disease virus in samples of experimentally infected cattle using different diagnostic techniques

2005 ◽  
Vol 72 (2) ◽  
Author(s):  
E.S.M. Tuppurainen ◽  
E.H. Venter ◽  
J.A.W. Coetzer
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Skin Disease ◽  
Virus Isolation ◽  
Skin Lesions ◽  
Viral Dna ◽  
Lumpy Skin Disease ◽  
Transmission Electron ◽  
Skin Biopsies ◽  
Skin Samples

Lumpy skin disease (LSD) is a disease of cattle, primarily in Africa and Madagascar and rarely in the Middle East. It is caused by a capripoxvirus that belongs to the family Poxviridae. The disease is of economic importance in endemic areas. Effective control of LSD requires accurate and rapid laboratory techniques to confirm a tentative clinical diagnosis. Comparative studies on different diagnostic tests used at different stages of the disease have not been done. The aim of this study was to compare several of these tests. Six seronegative bulls, between 11 and 20 months of age, were infected intravenously and kept in an insect-free facility. The course of the infection was monitored. During a 3-month period blood samples and skin biopsies were collected for virus isolation and polymerase chain reaction (PCR). Skin biopsies were also examined using transmission electron microscopy (TEM). The incubation period in infected animals varied from 4-5 days. The length of the viraemic period did not correlate with the severity of clinical disease. Viraemia was detected from 1-12 days using virus isolation and from 4-11 days using the PCR, which is longer than has previously been reported. Virus was isolated from skin biopsies until Day 39 post infection (p.i.) and PCR could demonstrate viral DNA until Day 92 p.i. Transmission electron microscopy of negatively stained skin biopsies detected LSD virus only in one of the four bulls that developed skin lesions until Day 33 p.i. The PCR was a fast and sensitive method to demonstrate viral DNA in blood and skin samples. It could detect viral nucleic acid in skin lesions 53 days longer than virus isolation. Virus isolation from blood and skin samples was sensitive and reliable, but as a single test it may be too time-consuming to use although this depends on how rapidly the diagnosis must be confirmed. In conclusion, this study showed the PCR to be superior in detecting LSD virus from blood and skin samples. However, virus isolation is still required when the infectivity of the LSD virus is to be determined. Indexed by Sabinet Online

scholarly journals Serum Biochemistry of Lumpy Skin Disease Virus-Infected Cattle

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Murat Şevik ◽  
Oğuzhan Avci ◽  
Müge Doğan ◽  
Ömer Barış İnce
Keyword(s):  
Real Time ◽  
Disease Virus ◽  
Real Time Pcr ◽  
Skin Disease ◽  
Infected Cattle ◽  
Serum Samples ◽  
Lumpy Skin Disease ◽  
Blood Samples ◽  
Lumpy Skin Disease Virus ◽  
Serum Biochemical

Lumpy skin disease is an economically important poxvirus disease of cattle. Vaccination is the main method of control but sporadic outbreaks have been reported in Turkey. This study was carried out to determine the changes in serum biochemical values of cattle naturally infected with lumpy skin disease virus (LSDV). For this study, blood samples in EDTA, serum samples, and nodular skin lesions were obtained from clinically infected animals (n=15) whereas blood samples in EDTA and serum samples were collected from healthy animals (n=15). A quantitative real-time PCR method was used to detectCapripoxvirus(CaPV) DNA in clinical samples. A real-time PCR high-resolution melt assay was performed to genotype CaPVs. Serum cardiac, hepatic, and renal damage markers and lipid metabolism products were measured by autoanalyzer. LSDV nucleic acid was detected in all samples which were obtained from clinically infected cattle. The results of serum biochemical analysis showed that aspartate aminotransferase, alkaline phosphatase, total protein, and creatinine concentrations were markedly increased in serum from infected animals. However, there were no significant differences in the other biochemical parameters evaluated. The results of the current study suggest that liver and kidney failures occur during LSDV infection. These findings may help in developing effective treatment strategies in LSDV infection.

scholarly journals Molecular detection of lumpy skin disease virus in cattle by polymerase chain reaction in Iraq

2016 ◽  
Vol 40 (1) ◽  
pp. 83-88 ◽  
Author(s):  
Khitam Mahdi Mhemid
Keyword(s):  
Polymerase Chain Reaction ◽  
Disease Virus ◽  
Skin Disease ◽  
Age Groups ◽  
Conventional Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Lumpy Skin Disease ◽  
Blood Samples ◽  
Lumpy Skin Disease Virus ◽  
Polymerase Chain

     This study is conducted to detect lumpy skin disease virus in Babylon, Al-Qadysia and Al-Muthana governorate during autumn 2014 using conventional polymerase chain reaction. A total of 150 specimens: 50 whole blood samples, 50 skin nodular biopsies and 50 tick samples were collected from infected animals of different breeds, genders and ages during lumpy skin disease outbreak. The results revealed that 104 cases (69.33%) were positive for lumpy skin disease virus by using polymerase chain reaction, with significant (P<0.05) differences between positive and negative cases. Out of 50 blood samples, 22 cases (44%) were positive for lumpy skin disease virus. The nodular skin samples collected from slaughtered animals showed 36 positive cases (72%), whereas 46 tick samples (92%) were positive for the disease, with significant (P<0.05) difference among them. According to gender, the finding showed significant results of lumpy skin disease in females (78.78%). It was recorded that higher percentage of positive cases was found in Friesian cattle (100%), crossbreed (73.58%) while native breed was (50.76%) with significant (P<0.05) difference among them. Regarding age groups, the results showed that all ages were susceptible to lumpy skin disease and significantly not different.

scholarly journals Molecular and histopathological confirmation of clinically diagnosed lumpy skin disease in cattle, Baghdad Province of Iraq

2019 ◽  
Vol 12 (11) ◽  
pp. 1826-1832
Author(s):  
Hasanain A. J. Gharban ◽  
Sattar J. J. Al-Shaeli ◽  
Hams H. H. Al-Fattli ◽  
Muthanna N. K. Altaee
Keyword(s):  
Clinical Examination ◽  
Skin Disease ◽  
Age Groups ◽  
Potential Method ◽  
Histopathological Analysis ◽  
Nasal Discharge ◽  
Hydropic Degeneration ◽  
Lumpy Skin Disease ◽  
Nodular Lesions ◽  
Skin Biopsies

Aim: This study aimed to confirm the clinically diagnosed cattle with lumpy skin disease (LSD) at Baghdad Province/Iraq from October 2018 to March 2019. Materials and Methods: Molecular polymerase chain reaction (PCR) and histopathology were applied for the detection of LSD among 71 infected cattle issued for slaughter. Results: Pre-slaughter clinical examination showed significant increases (p<0.05) in values of temperature (39.7±0.74°C), pulse (96.42±3.51), and respiratory (33.54±0.63) rates. Enlargement of lymph nodes (prescapular, supramammary, and prefemoral), lacrimation, mucopurulent nasal discharge, salivation, edema in limbs and head among severe infected cases, and marked fall in milk production was seen. An association of LSD to risk factors (age, gender, and areas) showed that there is significant elevation in prevalence of disease in >2-5 years (54.93%) rather than other age groups (>5 and <2 years) in females (73.24%) than males (26.76%); and in sub-rural (42.25%) and rural (39.44%) compared to urban (18.31%) areas. Postmortem examination appeared nodular lesions in upper parts of the digestive system (9.86%), rumen (2.82%), upper respiratory tracts (7.04%), and lung (4.23%). The PCR examination of P32 and thymidine kinase antigenic genes showed 90.14% and 60.56% positive samples, respectively. Histopathological analysis of nodular skin biopsies showed edema, hyperemia, acanthosis, severe hydropic degeneration, and hyperkeratosis in epidermis; whereas, mononuclear cell infiltration, inclusion bodies, and vasculitis seen in the dermis. Conclusion: PCR and histopathology assay could be a potential method to confirm the LSD infection concomitant with clinical examination.

scholarly journals Detection of lumpy skin disease virus in cattle using real-time polymerase chain reaction and serological diagnostic assays in different governorates in Egypt in 2017

2019 ◽  
Vol 12 (7) ◽  
pp. 1093-1100 ◽  
Author(s):  
Gamil Sayed Gamil Zeedan ◽  
Ayman Hamid Mahmoud ◽  
Abeer Mostafa Abdalhamed ◽  
Khaled Abd El-Hamid Abd El-Razik ◽  
Manal Hamdy Khafagi ◽  
...  
Keyword(s):  
Clinical Signs ◽  
Rt Pcr ◽  
Conventional Pcr ◽  
Serum Samples ◽  
Lumpy Skin Disease ◽  
Blood Samples ◽  
Diagnostic Assays ◽  
Cattle Serum ◽  
Skin Biopsies ◽  
Polymerase Chain

Background and Aim: Lumpy skin disease (LSD), is a highly infectious viral disease of cattle, caused by LSD virus (LSDV) which belongs to the genus Capripoxvirus of family Poxviridae. In the summer of 2017, skin lesions suggestive of LSD were observed in cattle at several governorates in Egypt. This study aimed to detect LSDV in cattle specimens using rapid serological and molecular diagnostic assays. Materials and Methods: A total of 46 skin biopsies and uncoagulated blood samples were collected from cattle with LSD suggestive clinical signs, as well as 290 coagulated whole blood samples from cattle without skin lesion in different governorates in Egypt during the summer of 2017. Skin biopsies were used for virus isolation from the chorioallantoic membrane of 11-day-old specific pathogen-free embryonated chicken eggs (SPF-ECEs). LSDV was identified using conventional polymerase chain reaction (PCR), real-time PCR (RT-PCR), and fluorescent antibody technique (FAT) with specific hyperimmune serum against LSDV. Cattle sera were examined using indirect FAT (IFAT) and indirect enzyme-linked immunosorbent assay (ELISA). Results: Skin nodules and sitfast lesions were significant clinical signs observed in all LSD suspect cattle. SPF-ECEs, from which positive isolations were made and it showed characteristic inflammatory and focal white pock lesions. The isolated viruses were identified as LSDV by FAT, conventional gel-based PCR, and RT-PCR. Among the skin biopsies and corresponding blood samples, LSDV-positive samples percentage were 39.13 and 36.95 by RT-PCR, followed 34.78 and 28.26 by conventional PCR and then 32.6 and 26.8 by FAT, respectively. The total positive percentage of LSDV antibody detected in cattle serum samples were 17.93 and 14.48 by indirect ELISA and IFAT. Conclusion: LSDV was detected and identified in skin biopsies and corresponding blood samples of naturally infected cattle, more LSDV-positive samples were detected by RT-PCR, followed by conventional PCR and then FAT. The indirect ELISA detected more antibody-positive samples than the IFAT from cattle serum samples. The RT-PCR assay is simple, sensitive, rapid, and reliable for the detection of LSDV in blood and skin nodule biopsies of suspected cattle.

scholarly journals Lumpy skin disease (LSD): Pathomorphological features and molecular detection in dairy cattle of west coastal India

2021 ◽  
Author(s):  
Shivasharanappa Nayakvadi ◽  
Samruddhi Prasad Joshi ◽  
Susitha Rajkumar ◽  
ChethanKumar HB ◽  
Jagruti Bathini ◽  
...  
Keyword(s):  
Skin Disease ◽  
Molecular Detection ◽  
Granulomatous Inflammation ◽  
Cattle Population ◽  
Vacuolar Degeneration ◽  
Lumpy Skin Disease ◽  
Blood Samples ◽  
P32 Gene ◽  
Pcr Targeting ◽  
Rpo30 Gene

Lumpy skin disease (LSD) is an emerging pox viral disease affecting cattle population worldwide. In India, the first outbreak of LSD is reported during August 2019 in Odisha state, which then followed by outbreaks in crossbred and indigenous cattle population of other states. Present investigation designed to study the prevalence, pathomorphological changes and molecular detection of LSD virus in naturally infected cattle. The overall morbidity of LSD was 4.48% among 30 dairy farms. Skin nodular biopsy, whole blood and serum samples (n= 66) were collected for the diagnosis of LSD by histopathology, PCR and sequencing. The envelope protein gene (P32), Fusion protein (F) and DNA dependent RNA polymerase 30 kDa subunit (RPO30) genes were targeted for PCR testing. Out of 66, 46 cattle showed generalized skin nodules and papules of various sizes (0.5 - 6.5cm) on the skin particularly at neck, face, nose, tail, perineum and udder. Microscopic examination of the skin nodule biopsy tissue revealed presence of diffuse granulomatous inflammation, hyperkeratosis, focal to diffuse vasculitis and lymphangitis, vacuolar degeneration, spongiosis and acanthosis. The inflammatory cells typically comprised of macrophages, lymphocytes, neutrophils and eosinophils along with diffuse necrosis in dermis in chronic cases. The eosinophilic intracytoplasmic viral inclusions in keratinocytes and epithelial cells were detected in few cases. Gel-PCR assay detected P32 gene in 83%, F gene in 72% and RPO30 gene in 77% of skin biopsy samples. Three blood samples were also found positive for P32 gene by PCR. Whereas TaqMan™ probe Real Time PCR targeting EEV glycoprotein gene (LSDV126) detected LSDV in 94% of biopsy samples and three blood samples which indicated its higher sensitive for the diagnosis of LSDV. Phylogenetic analysis of RPO30 gene sequence showed that the isolates from this study were grouped in same cluster with LSDV isolates of Bangladesh, Kenya and other Indian isolates detected during 2019-20.

scholarly journals Lumpy Skin Disease (LSD): Pathomorphological Features and Molecular Detection in Dairy Cattle of West Coastal India

2021 ◽  
Author(s):  
Shivasharanappa Nayakvadi ◽  
Samruddhi Prasad Joshi ◽  
Susitha Rajkumar ◽  
Chethan Kumar H. B. ◽  
Jagruti Bathini ◽  
...  
Keyword(s):  
Skin Disease ◽  
Molecular Detection ◽  
Granulomatous Inflammation ◽  
Cattle Population ◽  
Vacuolar Degeneration ◽  
Lumpy Skin Disease ◽  
Blood Samples ◽  
P32 Gene ◽  
Pcr Targeting ◽  
Rpo30 Gene

Abstract Lumpy skin disease (LSD) is an emerging pox viral disease affecting cattle population worldwide. In India, the first outbreak of LSD is reported during August 2019 in Odisha state, which then followed by outbreaks in crossbred and indigenous cattle population of other states. Present investigation designed to study the prevalence, pathomorphological changes and molecular detection of LSD virus in naturally infected cattle. The overall morbidity of LSD was 4.48% among 30 dairy farms. Skin nodular biopsy, whole blood and serum samples (n= 66) were collected for the diagnosis of LSD by histopathology, PCR and sequencing. The envelope protein gene (P32), Fusion protein (F) and DNA dependent RNA polymerase 30 kDa subunit (RPO30) genes were targeted for PCR testing. Out of 66, 46 cattle showed generalized skin nodules and papules of various sizes (0.5 - 6.5cm) on the skin particularly at neck, face, nose, tail, perineum and udder. Microscopic examination of the skin nodule biopsy tissue revealed presence of diffuse granulomatous inflammation, hyperkeratosis, focal to diffuse vasculitis and lymphangitis, vacuolar degeneration, spongiosis and acanthosis. The inflammatory cells typically comprised of macrophages, lymphocytes, neutrophils and eosinophils along with diffuse necrosis in dermis in chronic cases. The eosinophilic intracytoplasmic viral inclusions in keratinocytes and epithelial cells were detected in few cases. Gel-PCR assay detected P32 gene in 83%, F gene in 72% and RPO30 gene in 77% of skin biopsy samples. Three blood samples were also found positive for P32 gene by PCR. Whereas TaqMan™ probe Real Time PCR targeting EEV glycoprotein gene (LSDV126) detected LSDV in 94% of biopsy samples and three blood samples which indicated its higher sensitive for the diagnosis of LSDV. Phylogenetic analysis of RPO30 gene sequence showed that the isolates from this study were grouped in same cluster with LSDV isolates of Bangladesh, Kenya and other Indian isolates detected during 2019-20.

scholarly journals Molecular Characterization of Lumpy Skin Disease Virus Isolates from Outbreak Cases in Cattle from Sawena District of Bale Zone, Oromia, Ethiopia

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Shubisa Abera Leliso ◽  
Fufa Dawo Bari ◽  
Tesfaye Rufael Chibssa
Keyword(s):  
Skin Disease ◽  
Molecular Techniques ◽  
Effective Control ◽  
Economic Losses ◽  
Morbidity Rate ◽  
Lumpy Skin Disease ◽  
Significant Difference ◽  
Nasal Swabs ◽  
Skin Biopsies ◽  
Bale Zone

Lumpy skin disease (LSD) is a viral disease caused by LSD virus and is one of the most economically significant transboundary and emerging diseases of cattle. LSD causes considerable economic losses due to emaciation, damage to hides, infertility, and loss of milk production. In Ethiopia, the disease is distributed almost in all regions and is regarded as one of the most economically important livestock diseases in the country. An outbreak investigation of the disease was monitored from October 2016 to April 2017 in southern pastoral areas of Bale Zone, Oromia, Ethiopia. In December 2016, LSD outbreak occurred in Sawena district of Bale Zone, from which necessary biopsy samples were collected from actively infected animals for the purpose of virus isolation, and characterization using different molecular techniques at National Animal Health and Diagnostic Investigation Center (NAHDIC) of Sebeta, Ethiopia. In addition, clinical examination of infected and in-contact animals was carried out together with a questionnaire survey. Based on the clinical manifestations, LSD was recorded in 18% (94/522) of examined cattle, whereas biopsy samples from 20 clinically positive animals were collected for further laboratory process. The morbidity rate was higher in animals less than two years 28.97% (31/107) than other ages and showed a statistically significant difference with P < 0.05 . Female animals showed higher morbidity rate of 20.59% (76/369) than male animals (11.76%) (18/153) with a significant difference at P ≤ 0.003 . Mortality rate and case fatality were also significantly higher in young animals than other age groups. Viruses were isolated from both skin biopsies and nasal swabs on Vero cell line. From both skin biopsies and nasal swabs, the virus DNA was identified by amplifying the 172 bp DNA fragment using real-time and conventional PCR. Providing adequate diagnostic facilities, establishing strategic policies for effective control and eradication and awareness creations for communities for early identification or reporting were recommendations made to minimize economic losses of the disease.

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