scholarly journals First molecular confirmation of Coenurus cerebralis in sheep and goats with neurological behaviors in Iraq

2021 ◽  
pp. 1420-1425
Author(s):  
Eva Aisser Ajaj ◽  
Hadeel Asim Mohammad ◽  
Hasanain A. J. Gharban
Keyword(s):  
Historical Data ◽  
Intermediate Stage ◽  
Molecular Techniques ◽  
Economic Losses ◽  
Sheep And Goats ◽  
Neurological Signs ◽  
Coenurus Cerebralis ◽  
Polymerase Chain ◽  
Female Sheep

Background and Aim: Coenurus cerebralis is the intermediate stage of the canine cestode, which infect sheep and goats, resulting mainly in neurological signs and causing direct and indirect economic losses. This study aimed to demonstrate the existence of C. cerebralis and to elucidate the role of this parasite in inducing neurological behaviors in sheep and goats. Materials and Methods: On the basis of historical data on neurological signs, we subjected 76 animals (49 sheep and 27 goats) of different ages, sexes, and geographical areas for molecular examination of their blood samples using the polymerase chain reaction assay. Results: Of the 76 animals, 23.68% tested positive for C. cerebralis infection. We found significant increases in infection (p<0.05) in sheep (26.53%) more than in goats (18.52%). Circling movement was prevalent significantly among both C. cerebralis-positive sheep and goats. The Nineveh region had a significant (p<0.05) increase in positive sheep and goats, and the sheep of all study regions were infected significantly (p<0.05) more than were the goats. We found no significant (p>0.05) variation between sheep ages ≥1-3 years and ≥3 years; however, both groups had a significantly (p≤0.043) higher positivity rate than did sheep ages <1 year. The findings of sheep ages <1 year and ≥1-3 years were significantly higher than those of the goats, but not for goats ages ≥3 years. Female sheep and goats showed a significant increase in positivity versus that for the males. Conclusion: To the best of our knowledge, this study is the first report in Iraq targeting detection of C. cerebralis in sheep and goats with neurological behaviors; therefore, additional studies involving different animals in other regions using molecular techniques are needed.

scholarly journals Molecular detection of cattle Sarcocystis spp. in North-West Italy highlights their association with bovine eosinophilic myositis

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Selene Rubiola ◽  
Tiziana Civera ◽  
Felice Panebianco ◽  
Davide Vercellino ◽  
Francesco Chiesa
Keyword(s):  
18S Rdna ◽  
Inflammatory Myopathy ◽  
Molecular Techniques ◽  
Diaphragm Muscle ◽  
Economic Losses ◽  
Intermediate Hosts ◽  
Slaughter Cattle ◽  
Significant Difference ◽  
Pcr Targeting

Abstract Background Cattle are intermediate hosts of six Sarcocystis species, among which Sarcocystis hominis and Sarcocystis heydorni can infect humans through the consumption of raw or undercooked meat. In addition to the zoonotic potential, there is increasing interest in these protozoa because of the evidence supporting the role of Sarcocystis spp. in the occurrence of bovine eosinophilic myositis (BEM), a specific inflammatory myopathy which leads to carcass condemnation and considerable economic losses. Actually, all the prevalence studies carried out on cattle in Italy have been based on either morphological or 18S rDNA-based molecular techniques, most likely leading to misidentification of closely related species. Therefore, there is a strong need for new data on the prevalence of the different Sarcocystis spp. in cattle in Italy and their association with bovine eosinophilic myositis. Methods To reach our aim, individual striated muscle samples from BEM condemned carcasses (N = 54) and diaphragm muscle samples from randomly sampled carcasses (N = 59) were obtained from Northwest Italy slaughterhouses. Genomic DNA was extracted and analyzed by multiplex-PCR targeting 18S rDNA and cox1 genes. PCR products amplified using the genus-specific primer set in absence of the specific fragment for S. hirsuta, S. cruzi, S. hominis or S. bovifelis were sequenced to achieve species identification. Results Sarcocystis DNA was detected in 67.8% of the samples from slaughter cattle and in 90.7% of the samples from BEM condemned carcasses. S. cruzi was identified as the most prevalent species in slaughter cattle (61%), followed by S. bovifelis (10.2%), S. hominis (8.5%) and S. hirsuta (1.7%). Notably, among the different Sarcocystis spp. detected, the presence of S. bovifelis and S. hominis was significantly higher in samples isolated from BEM condemned carcasses (46.3% and 40.7% respectively), while there was no statistically significant difference between the presence of S. cruzi or S. hirsuta in BEM condemned carcasses (42.6% and 1.8%, respectively) and randomly sampled carcasses. Furthermore, DNA sequence analysis revealed the presence of a putative new species in two carcasses. Conclusions Our study contributes to updating the data on the prevalence of the different Sarcocystis spp. in cattle in Italy, highlighting the presence of three Sarcocystis spp., S. cruzi, S. hominis and S. bovifelis, in BEM lesions and allowing us to speculate on the possible role of S. hominis and S. bovifelis as the major sarcosporidian species involved in bovine eosinophilic myositis. Graphic Abstract

scholarly journals Sensitive SYBR Green—Real Time PCR for the Detection and Quantitation of Avian Rotavirus A

2018 ◽  
Vol 6 (1) ◽  
pp. 2 ◽  
Author(s):  
David De la Torre ◽  
Claudete Astolfi-Ferreira ◽  
Ruy Chacon ◽  
Antonio Piantino Ferreira
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Sybr Green ◽  
Molecular Techniques ◽  
Economic Losses ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Rotavirus A ◽  
Avian Nephritis Virus ◽  
Polymerase Chain

Avian rotavirus A (ARtV-A) is a virus that affects young birds, causing acute diarrhea and economic losses in the poultry industry worldwide. The techniques used for the diagnosis of ARtV-A include electron microscopy, isolation in cell culture, and serology, as well as molecular techniques, such as the reverse transcription-polymerase chain reaction (RT-PCR). The objective of this work was to standardize a real-time RT-polymerase chain reaction (RT-qPCR) using SYBR Green chemistry for the rapid detection and quantification of ARtV-A from bird tissues and materials fixed on FTA cards on the basis of the nucleotide sequence of segment 6 (S6), which codes for the structural VP6 protein of ARtV-A. The results show the efficient amplification of the proposed target, with a limit of detection (LoD) of one copy gene (CG) per microliter of cDNA and a limit of quantification (LoQ) of 10 CGs per microliter. The efficiency of the primers was determined to be 95.66% using a standard curve, with an R2 value of 0.999 and a slope of −3.43. The specificity was determined using samples coinfected with ARtV-A, the chicken parvovirus, the chicken astrovirus, and the avian nephritis virus as positive controls and commercially available vaccines of the infectious bronchitis virus, infectious bursa disease virus, avian reovirus and healthy organs as negative controls. This technique, which lacks nonspecific PCR products and dimers, demonstrated greater sensitivity and specificity than conventional RT-PCR, and it reduced the analysis time by more than 50%.

scholarly journals Brucella species circulating in wildlife in Serengeti ecosystem, Tanzania

2020 ◽  
Author(s):  
R. M. Sambu ◽  
C. Mathew ◽  
H.E. Nonga ◽  
A. S. Lukambagire ◽  
Richard B. Yapi ◽  
...  
Keyword(s):  
Insertion Sequence ◽  
Surface Protein ◽  
Molecular Techniques ◽  
Cross Sectional ◽  
Serengeti Ecosystem ◽  
High Interaction ◽  
Polymerase Chain ◽  
Shed Light ◽  
Edta Blood

AbstractBackgroundBrucellosis is a bacterial zoonosis of public health and economic importance world-wide. It affects a number of domestic animals, wildlife and humans. This study was carried out to determine circulating Brucella species in wildlife in Serengeti ecosystem using molecular techniques.MethodologyA total of 189 samples including EDTA blood, serum and amniotic fluid from buffalos, lions, wildebeest, impala, zebra and hyena that were collected in relation to different cross-sectional studies conducted in the Serengeti ecosystem in Tanzania were used. Multiplex polymerase chain reaction AMOS-PCR and quantitative Real-Time PCR (qPCR) targeting the genus specific surface protein bcsp31 gene and the insertion sequence IS711 element downstream of the alkB gene for B. abortus and BMEI1162 gene for B. melitensis were employed on the samples.ResultsResults indicated that out of 189 samples examined, 12 (6.4%) and 22 (11.6%) contained Brucella DNA as detected by AMOS-PCR and qPCR, respectively. Most of the positive samples were from lions (52.6%) and buffaloes (19.6%). Other animals that were positive included wildebeest, impala, zebra and hyena. Out of 22 positive samples, 16 (66.7%) were identified as B. abortus and the rest were B. melitensis.ConclusionDetection of zoonotic Brucella species in wildlife suggests that livestock and humans at the interface areas where there is high interaction are at risk of acquiring the infection. Therefore, public education to interrupt risky transmission practices is needed. The findings also shed light on the transmission dynamics around interface areas and the role of wildlife in transmission and maintenance of Brucella infection in the region.

scholarly journals Molecular-based cross-species evaluation of bovine coronavirus infection in cattle, sheep and goats in Ghana.

2020 ◽  
Author(s):  
Vitus Burimuah ◽  
Augustina Sylverken ◽  
Michael Owusu ◽  
Philip El-Duah ◽  
Richmond Yeboah ◽  
...  
Keyword(s):  
Clinical Signs ◽  
Small Ruminants ◽  
Economic Losses ◽  
Sample Collection ◽  
Bovine Coronavirus ◽  
Sheep And Goats ◽  
Homologous Sequences ◽  
Rdrp Region ◽  
Coronavirus Infection ◽  
Polymerase Chain

Abstract Background: Apart from the huge worldwide economic losses often occasioned by bovine coronavirus (BCoV) to the livestock industry particularly cattle, continuous surveillance of the virus in cattle and small ruminants is essential in monitoring variations in the virus that could enhance host switching. In this study, we collected rectal swabs from a total of 1,498 cattle, sheep and goats. BCoV detection was based on reverse transcriptase polymerase chain reaction. Sanger sequencing of the partial RNA-dependent RNA polymerase (RdRp) region for postive samples were done and nucleotide sequences were compared with homologous sequences from the GenBank.Results: The study reports a BCoV prevalence of 0.3% consisting of 4 positive cases; 3 goats and 1 cattle. Less than 10% of all the animals sampled showed clinical signs such as diarrhea and respiratory distress except for high temperature which occurred in > 1000 of the animals. However, none of the 4 BCoV positive animals manifested any clinical signs of the infection at the time of sample collection. Bayesian majority-rule cladogram comparing partial and full length BCoV RdRp genes obtained in the study to data from the GenBank revealed that the sequences obtained from this study formed one large monophyletic group with those from different species and countries. The goat sequences were similar to each other and clustered within the same clade. No major variations were thus observed with our isolates and those from elsewhere.Conclusion: Given that Ghana predominantly practice the extensive and semi-intensive systems of animal rearing, our study highlights the potential for spillover of BCoV to small ruminants in settings with mixed husbandry and limited separation between species.

scholarly journals Molecular-Based Cross-Species Evaluation of Bovine Coronavirus Infection in Cattle, Sheep and Goats in Ghana.

2020 ◽  
Author(s):  
Vitus Burimuah ◽  
Augustina Sylverken ◽  
Michael Owusu ◽  
Philip El-Duah ◽  
Richmond Yeboah ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Small Ruminants ◽  
Economic Losses ◽  
Rna Dependent Rna Polymerase ◽  
Bovine Coronavirus ◽  
Sheep And Goats ◽  
Homologous Sequences ◽  
Rdrp Region ◽  
Coronavirus Infection ◽  
Polymerase Chain

Abstract Background: Apart from the huge worldwide economic losses often occasioned by bovine coronavirus (BCoV) to the livestock industry particularly cattle, continuous surveillance of the virus in cattle and small ruminants is essential in monitoring variations in the virus that could enhance host switching.Methods: In this study, we collected rectal swabs from a total of 1,498 cattle, sheep and goats. BCoV detection was based on reverse transcriptase polymerase chain reaction. Sanger sequencing of the partial RNA-dependent RNA polymerase (RdRp) region for postive samples were done and nucleotide sequences were compared with homologous sequences from the GenBank.Results: The study reports a BCoV prevalence of 0.3% consisting of 4 positive cases; 3 goats and 1 cattle. Bayesian majority-rule cladogram comparing partial and full length BCoV RNA-dependent RNA polymerase (RdRp) genes obtained in the study to data from the GenBank revealed that the sequences obtained from this study formed one large monophyletic group with those from different species and countries. The goat sequences were similar to each other and clustered within the same clade. No major variations were thus observed with our isolates and those from elsewhere.Conclusion: Given that Ghana predominantly practice the extensive and semi-intensive systems of animal rearing, our study highlights the potential for spillover of BCoV to small ruminants in settings with mixed husbandry and limited separation between species. Continuous surveillance of BCoV in the country is thus critical to avert future surge and probable increase in host range.

scholarly journals A Hypothesis for Explaining the Geographical Distribution of Crohn’s Disease

1996 ◽  
Vol 10 (3) ◽  
pp. 173-177 ◽  
Author(s):  
Cyrus P Tamboli
Keyword(s):  
Polymerase Chain Reaction ◽  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Geographical Distribution ◽  
Environmental Influence ◽  
Molecular Techniques ◽  
Epidemiological Evidence ◽  
Chain Reaction ◽  
Polymerase Chain

The etiology of Crohn’s disease (CD) remains unknown, although there is epidemiological evidence supporting an environmental influence. Recent molecular techniques, including polymerase chain reaction, have renewed interest in a possible etiological role ofMycobacterium paratuberculosis, which has been isolated from a number of CD patients. The organism causes a chronic enteritis in animals called paratuberculosis, a condition with many clinical and pathological similarities to CD. This review compares the epidemiology of paratuberculosis in animals with the epidemiology of CD in humans. There is considerable overlap of regions with high prevalences of paratuberculosis and CD. This finding adds support to the implication ofM paratuberculosisin the etiology of CD.

What Is the Role of Molecular Techniques for Detection of Pathogen Deoxyribonucleic Acid (DNA) (Polymerase Chain Reaction [PCR] or Next-Generation Sequencing) in Patients With Infected Total Ankle Arthroplasty (TAA)?

2019 ◽  
Vol 40 (1_suppl) ◽  
pp. 29S-31S
Author(s):  
Khaled Emara ◽  
Amiethab Aiyer ◽  
Ryan Rogero
Keyword(s):  
Next Generation Sequencing ◽  
Clinical Evidence ◽  
Molecular Techniques ◽  
Total Ankle Arthroplasty ◽  
Next Generation ◽  
Level Of Evidence ◽  
Ankle Arthroplasty ◽  
Polymerase Chain ◽  
Generation Sequencing

Recommendation: Molecular techniques, particularly next-generation sequencing and the Ibis T5000 technology, have the potential to be used as an important adjunct in the diagnosis of bacterial infection following total ankle arthroplasty (TAA), although sufficient clinical evidence is lacking. Level of Evidence: Limited. Delegate Vote: Agree: 100%, Disagree: 0%, Abstain: 0% (Unanimous, Strongest Consensus)

scholarly journals Molecular Detection of Cattle Sarcocystis Spp. In North-West Italy Targeting Cox1 And 18S Genes Highlights Its Association With Bovine Eosinophilic Myositis.

2021 ◽  
Author(s):  
Selene Rubiola ◽  
Tiziana Civera ◽  
Felice Panebianco ◽  
Davide Vercellino ◽  
Francesco Chiesa
Keyword(s):  
Inflammatory Myopathy ◽  
Molecular Techniques ◽  
Economic Losses ◽  
Intermediate Hosts ◽  
Prevalence Studies ◽  
North West ◽  
Slaughter Cattle ◽  
Significant Difference ◽  
Pcr Targeting

Abstract Background: Cattle are intermediate hosts of six Sarcocystis species, among which Sarcocystis hominis and Sarcocystis heydorni can infect humans through the consumption of raw or undercooked meat. In addition to the zoonotic potential, there is increasing interest in these protozoa due to the evidences supporting the role of Sarcocystis spp. in the occurrence of bovine eosinophilic myositis (BEM), a specific inflammatory myopathy which leads to carcass condemnation and considerable economic losses. Actually, all the prevalence studies carried out on cattle in Italy have been based either on morphological or 18S rDNA-based molecular techniques, most likely leading to misidentification of closely related species. Therefore, there is a strong need for new data on the prevalence of the different Sarcocystis spp. in cattle in Italy and their association with bovine eosinophilic myositis.Methods: To reach our aim, individual cattle samples from BEM condemned carcasses (N=54) and randomly sampled carcasses (N=59) were obtained from Piedmont slaughterhouses. Genomic DNA was extracted and analyzed by multiplex-PCR targeting 18S and cox1 genes. PCR products amplified using the genus specific primer set in absence of the specific fragment for S. hirsuta, S. cruzi, S. hominis or S. bovifelis, were sequenced to achieve species identification.Results: Sarcocystis DNA was detected in 67.8% of the samples from slaughter cattle and in 90.7% of the samples from BEM condemned carcasses. S. cruzi was identified as the most prevalent species (61%), followed by S. bovifelis (10.2%), S. hominis (8.5%) and S. hirsuta (1.7%). Notably, among the different Sarcocystis spp. detected, the presence of S. bovifelis and S. hominis was significantly higher in samples isolated from BEM condemned carcasses (46.3% and 40.7% respectively), while there was no statistically significant difference between the presence of S. cruzi or S. hirsuta in BEM condemned carcasses and randomly sampled carcasses. Furthermore, DNA sequence analysis revealed the presence of a putative new species in 2 carcasses.Conclusions: Our study contributes to update the data on the prevalence of the different Sarcocystis spp. in cattle in Italy and emphasize the role of S. hominis and S. bovifelis as the major sarcosporidian species involved.

scholarly journals Molecular-based cross-species evaluation of bovine coronavirus infection in cattle, sheep and goats in Ghana.

2020 ◽  
Author(s):  
Vitus Burimuah ◽  
Augustina Sylverken ◽  
Michael Owusu ◽  
Philip El-Duah ◽  
Richmond Yeboah ◽  
...  
Keyword(s):  
Clinical Signs ◽  
Small Ruminants ◽  
Economic Losses ◽  
Sample Collection ◽  
Bovine Coronavirus ◽  
Sheep And Goats ◽  
Homologous Sequences ◽  
Rdrp Region ◽  
Coronavirus Infection ◽  
Polymerase Chain

Abstract Background: Apart from the huge worldwide economic losses often occasioned by bovine coronavirus (BCoV) to the livestock industry particularly cattle, continuous surveillance of the virus in cattle and small ruminants is essential in monitoring variations in the virus that could enhance host switching. In this study, we collected rectal swabs from a total of 1,498 cattle, sheep and goats. BCoV detection was based on reverse transcriptase polymerase chain reaction. Sanger sequencing of the partial RNA-dependent RNA polymerase (RdRp) region for postive samples were done and nucleotide sequences were compared with homologous sequences from the GenBank.Results: The study reports a BCoV prevalence of 0.3% consisting of 4 positive cases; 3 goats and 1 cattle. Less than 10% of all the animals sampled showed clinical signs such as diarrhea and respiratory distress except for high temperature which occurred in > 1000 of the animals. However, none of the 4 BCoV positive animals manifested any clinical signs of the infection at the time of sample collection. Bayesian majority-rule cladogram comparing partial and full length BCoV RdRp genes obtained in the study to data from the GenBank revealed that the sequences obtained from this study formed one large monophyletic group with those from different species and countries. The goat sequences were similar to each other and clustered within the same clade. No major variations were thus observed with our isolates and those from elsewhere.Conclusion: Given that Ghana predominantly practice the extensive and semi-intensive systems of animal rearing, our study highlights the potential for spillover of BCoV to small ruminants in settings with mixed husbandry and limited separation between species.

scholarly journals Molecular-based cross-species evaluation of bovine coronavirus infection in cattle, sheep and goats in Ghana

2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Vitus Burimuah ◽  
Augustina Sylverken ◽  
Michael Owusu ◽  
Philip El-Duah ◽  
Richmond Yeboah ◽  
...  
Keyword(s):  
Clinical Signs ◽  
Small Ruminants ◽  
Economic Losses ◽  
Sample Collection ◽  
Bovine Coronavirus ◽  
Sheep And Goats ◽  
Rdrp Region ◽  
Coronavirus Infection ◽  
Polymerase Chain ◽  
Cattle Rearing

Abstract Background Apart from the huge worldwide economic losses often occasioned by bovine coronavirus (BCoV) to the livestock industry, particularly with respect to cattle rearing, continuous surveillance of the virus in cattle and small ruminants is essential in monitoring variations in the virus that could enhance host switching. In this study, we collected rectal swabs from a total of 1,498 cattle, sheep and goats. BCoV detection was based on reverse transcriptase polymerase chain reaction. Sanger sequencing of the partial RNA-dependent RNA polymerase (RdRp) region for postive samples were done and nucleotide sequences were compared with homologous sequences from the GenBank. Results The study reports a BCoV prevalence of 0.3%, consisting of 4 positive cases; 3 goats and 1 cattle. Less than 10% of all the animals sampled showed clinical signs such as diarrhea and respiratory distress except for high temperature which occurred in > 1000 of the animals. However, none of the 4 BCoV positive animals manifested any clinical signs of the infection at the time of sample collection. Bayesian majority-rule cladogram comparing partial and full length BCoV RdRp genes obtained in the study to data from the GenBank revealed that the sequences obtained from this study formed one large monophyletic group with those from different species and countries. The goat sequences were similar to each other and clustered within the same clade. No major variations were thus observed between our isolates and those from elsewhere. Conclusions Given that Ghana predominantly practices the extensive and semi-intensive systems of animal rearing, our study highlights the potential for spillover of BCoV to small ruminants in settings with mixed husbandry and limited separation between species.

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