scholarly journals Isolation and Cultivation of a New Isolate of BTV-25 and Presumptive Evidence for a Potential Persistent Infection in Healthy Goats

Viruses ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 983
Author(s):  
Christina Ries ◽  
Ursula Domes ◽  
Britta Janowetz ◽  
Jens Böttcher ◽  
Katinka Burkhardt ◽  
...  
Keyword(s):  
Clinical Signs ◽  
Small Ruminants ◽  
Full Genome ◽  
Serum Samples ◽  
Blood Samples ◽  
Experimental Inoculation ◽  
Serological Surveillance ◽  
First Time ◽  
Goat Flock ◽  
Edta Blood

Recently, several so-called “atypical” Bluetongue virus (BTV) serotypes were discovered, including BTV-25 (Toggenburg virus), in Switzerland. Most “atypical” BTV were identified in small ruminants without clinical signs. In 2018, two goats from a holding in Germany tested positive for BTV-25 genome by RT-qPCR prior to export. After experimental inoculation of the two goats with the BTV-25 positive field blood samples for generation of reference materials, viremia could be observed in one animal. For the first time, the BTV-25-related virus was isolated in cell culture from EDTA-blood and the full genome of isolate “BTV-25-GER2018” could be generated. BTV-25-GER2018 was only incompletely neutralized by ELISA-positive sera. We could monitor the BTV-25 occurrence in the respective affected goat flock of approximately 120 goats over several years. EDTA blood samples were screened with RT-qPCR using a newly developed BTV-25 specific assay. For serological surveillance, serum samples were screened using a commercial cELISA. BTV-25-GER2018 was detected over 4.5 years in the goat flock with intermittent PCR-positivity in some animals, and with or without concomitantly detected antibodies since 2015. We could demonstrate the viral persistence of BTV-25-GER2018 in goats for up to 4.5 years, and the first BTV-25 isolate is now available for further characterization.

scholarly journals Putative Novel Atypical BTV Serotype ‘36’ Identified in Small Ruminants in Switzerland

Viruses ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 721
Author(s):  
Christina Ries ◽  
Andrea Vögtlin ◽  
Daniela Hüssy ◽  
Tabea Jandt ◽  
Hansjörg Gobet ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
Clinical Symptoms ◽  
Clinical Signs ◽  
Virus Transmission ◽  
Small Ruminants ◽  
Facial Oedema ◽  
The Novel ◽  
Transmission Pathways ◽  
Goat Flock ◽  
Edta Blood

We identified a putative novel atypical BTV serotype ‘36’ in Swiss goat flocks. In the initial flock clinical signs consisting of multifocal purulent dermatitis, facial oedema and fever were observed. Following BTV detection by RT-qPCR, serotyping identified BTV-25 and also a putative novel BTV serotype in several of the affected goats. We successfully propagated the so-called “BTV-36-CH2019” strain in cell culture, developed a specific RT-qPCR targeting Segment 2, and generated the full genome by high-throughput sequencing. Furthermore, we experimentally infected goats with BTV-36-CH2019. Regularly, EDTA blood, serum and diverse swab samples were collected. Throughout the experiment, neither fever nor clinical disease was observed in any of the inoculated goats. Four goats developed BTV viremia, whereas one inoculated goat and the two contact animals remained negative. No viral RNA was detected in the swab samples collected from nose, mouth, eye, and rectum, and thus the experimental infection of goats using this novel BTV serotype delivered no indications for any clinical symptoms or vector-free virus transmission pathways. The subclinical infection of the four goats is in accordance with the reports for other atypical BTVs. However, the clinical signs of the initial goat flock did most likely not result from infection with the novel BTV-36-CH0219.

scholarly journals Botulism (type A) in a horse - case report

2016 ◽  
Vol 85 (1) ◽  
pp. 71-76 ◽  
Author(s):  
Sevim Kasap ◽  
Hasan Batmaz ◽  
Meric Kocaturk ◽  
Frank Gessler ◽  
Serkan Catık ◽  
...  
Keyword(s):  
Botulinum Neurotoxin ◽  
Clinical Signs ◽  
Type A ◽  
Serum Samples ◽  
Specific Antibodies ◽  
Thoroughbred Horse ◽  
Botulinum Neurotoxin Type A ◽  
Botulinum Neurotoxins ◽  
First Time ◽  
Colonic Content

This paper presents the case of a six year-old, male, thoroughbred horse with clinical signs of inappetence, weakness, and incoordination when walking. Clinical examination showed that the horse staggered and leaned to the left side. Feedstuff was present inside and around its mouth. Salivation was increased and there was no reflex at the palpebrae and tongue. The horse had difficulty swallowing and the tone of its tail was reduced. Botulism was diagnosed based on the clinical signs. Antibiotic (ceftiofur) and fluid-electrolyte treatment was commenced. Next day, neostigmin was added to the horse’s treatment, and it became recumbent. The horse’s palpebral, tongue and tail reflexes returned partially after neostigmine methylsulphate treatment on the same day and it stood up on day four. However, it could not swallow anything during the whole week, so after getting permission from the owner, the horse was euthanized on day 10. Samples of the colonic content and blood serum were sent by courier to the laboratory for toxin neutralization, however, botulinum neurotoxins could not be detected. After that, serum samples from days 6 and 10 were sent to another laboratory for testing for botulinum neurotoxin antibodies by ELISA. Specific antibodies against botulinum neurotoxin type A were measured, indicating a previous, immuno-relevant contact with the toxin. This seroconversion for type A supports the clinical botulism diagnosis. Type A botulism is rarely seen in Europe and has been detected in a horse in Turkey for the first time.

scholarly journals Reinfection with Anaplasma phagocytophilum in BALB/c Mice and Cross-Protection between Two Sympatric Isolates

2004 ◽  
Vol 72 (8) ◽  
pp. 4723-4730 ◽  
Author(s):  
Michael L. Levin ◽  
Dondrae J. Coble ◽  
Danielle E. Ross
Keyword(s):  
Immune Response ◽  
Quantitative Pcr ◽  
Anaplasma Phagocytophilum ◽  
Ixodes Scapularis ◽  
Domestic Animals ◽  
Serum Samples ◽  
Partial Protection ◽  
Blood Samples ◽  
Or Groups ◽  
First Time

ABSTRACT Infection with Anaplasma phagocytophilum in white-footed mice results in partial protection against reinfection with the same agent. However, humans and domestic animals may be sequentially exposed to different isolates of the agent circulating in the same or adjacent foci. We investigated whether immune response to a tick-borne infection with A. phagocytophilum provides protection against homologous and heterologous challenges. BALB/c mice were infected with one of the two sympatric isolates of A. phagocytophilum via tick bite and challenged 16 weeks later by Ixodes scapularis nymphs infected with either the same or the alternative isolate. As controls, groups of infected mice were challenged by uninfected ticks to confirm an absence of reactivation of the original infection or groups of naive mice were fed upon by ticks from cohorts used for an infectious challenge. Xenodiagnostic I. scapularis larvae were fed upon each mouse at 14 and 21 days postchallenge (PCH) and tested for the presence of A. phagocytophilum as freshly molted nymphs. Blood samples for quantitative PCR were collected at 7, 14, 21, and 70 days PCH. Serum samples were collected weekly to monitor development of immune response. The proportion of infected animals, levels of bacteremia, and the prevalence of infection in xenodiagnostic ticks were higher in groups of control mice exposed to A. phagocytophilum for the first time than in mice reinfected with either homologous or heterologous isolates. The presence of antibodies against A. phagocytophilum did not protect mice from a challenge with either homologous or heterologous isolates, however the ensuing reinfection was significantly milder and of a shorter duration than the first infection with either isolate.

scholarly journals Follow-up of dairy cattle naturally infected by Trypanosoma vivax after treatment with isometamidium chloride

2021 ◽  
Vol 30 (1) ◽  
Author(s):  
Kayo José Garcia de Almeida Castilho Neto ◽  
Ana Beatriz da Cruz Favaro Garcia ◽  
Otavio Luiz Fidelis Junior ◽  
Walter Beterquini Nagata ◽  
Marcos Rogério André ◽  
...  
Keyword(s):  
Dairy Cattle ◽  
Clinical Signs ◽  
Parasite Clearance ◽  
Disease Diagnosis ◽  
Trypanosoma Vivax ◽  
Serum Samples ◽  
Diminazene Aceturate ◽  
Bovine Trypanosomosis ◽  
Edta Blood

Abstract Trypanosoma vivax infections cause nonspecific clinical signs in cattle associated with aparasitemic intervals, making disease diagnosis a challenge. In Brazil, diminazene aceturate and isometamidium chloride (ISM) are available to treat bovine trypanosomosis. The objective of this study was to follow-up, by molecular and serological techniques, dairy cattle naturally infected by T. vivax after ISM treatment. Thirty cattle naturally infected with T. vivax received two applications of ISM, at a dosage of 1.0 mg/kg intramuscularly, on days 0 and 150. For T. vivax diagnosis, EDTA-blood and serum samples were evaluated on 0, 7, 15, 30, 60, 90, 120, 150, 180, 210, and 240 days after treatment PCR, Loop-mediated isothermal amplification (LAMP) and ELISA. Animals with persistent detection of T. vivax DNA by both PCR and LAMP were found and continuous detection of anti-T. vivax IgG antibodies by ELISA, suggesting the presence of T. vivax resistance to ISM. The combination of LAMP and ELISA tests can prevent misdiagnosis of the parasite clearance in treated cattle, contributing to better disease control. This is the first experiment that demonstrates the persistence infection of T. vivax under ISM treatment in a natural infected herd and evidence of ISM chemotherapy-resistant T. vivax in Brazil.

scholarly journals Molecular detection of bovine hemotrophic mycoplasmas in Mexico

2020 ◽  
Vol 13 (52) ◽  
pp. 67-82
Author(s):  
Rosa Estela Quiroz Castañeda ◽  
Kytzya Mejía Aragón ◽  
Hugo Aguilar Diaz ◽  
Jesús Francisco Preciado de la Torre
Keyword(s):  
Molecular Detection ◽  
Detection Method ◽  
Animal Health ◽  
Clinical Signs ◽  
Health Condition ◽  
Duplex Pcr ◽  
Blood Samples ◽  
End Point ◽  
Mycoplasma Wenyonii ◽  
First Time

The presence of hemoplasmas Candidatus Mycoplasma haemobos and Mycoplasma wenyonii that infect bovine cattle has been reported during the last years. Hemoplasmas may affect animal health either alone or in coinfections with other microorganisms, resulting in anemia and other clinical signs. In Mexico, only Ca. M. haemobos has been detected in cattle; in this work, we report for the first time in our country the presence of M. wenyonii in animals from different geographical sources amd we detected both hemoplasmas by duplex PCR. Also, by single end-point PCR, we found Ca. M. haemobos and M. wenyonii in 96% and 96.29% of the blood samples, respectively. Both hemoplasmas were detected in 50% of the samples analyzed, which suggest that the duplex PCR developed in this work might improve if some modifications are performed. This molecular detection method will provide valuable information to know the health condition of national cattle to prevent pathogen dispersion.

Investigations on First Confirmed Outbreak of Ovine Theileriosis (Theileria luwenshuni) from Maharashtra State, India

2020 ◽  
Author(s):  
V.S. Dhaygude ◽  
K. Kundu ◽  
B.P. Kamdi ◽  
U.R. Bagal ◽  
S.B. Bhosale ◽  
...  
Keyword(s):  
18S Rrna ◽  
Clinical Signs ◽  
Specific Treatment ◽  
Small Ruminants ◽  
18S Rrna Gene ◽  
Pcr Detection ◽  
Nucleotide Sequencing ◽  
Rrna Gene ◽  
Blood Samples ◽  
Theileria Luwenshuni

Background: Clinical theileriosis of small ruminants is tick-borne disease caused by Theileria lestoquardi, Theileria uilenbergi and Theileria luwenshuni. Theileria annulata, the causative agent of bovine tropical theileriosis in cattle, can also infect sheep but does not cause any significant illness. It is one of the economically important diseases. There are no reports of ovine clinical theileriosis from Maharashtra state and there is paucity of information on its epidemiology. This paper reports first confirmed outbreak of ovine theileriosis based on clinical signs, microscopic examination, PCR and sequencing in the Maharashtra State of India. Methods: Whole blood samples from 22 ailing sheep were collected and subjected to hematological examination. Blood smears stained with Leishman’s stain were examined under 100X objective of the microscope. The blood samples from sheep found positive by microscopic method were subjected to PCR detection of 18S rRNA gene of hemoprotozoa and then for nucleotide sequencing and sequence analysis.Conclusion: Samples from 14 out of 22 sheep were found positive for piroplasms of Theileria spp by light microscopy. All positive samples were further confirmed by PCR detection of 18S rRNA gene of hemoprotozoa. PCR amplification yielded expected product of 1750 bp for all samples. BLAST and phylogenetic analysis of one sample revealed high sequence homology with T. luwenshuni reported from India and other countries. Characteristic clinical signs like fever, progressive anaemia, laboured breathing, lymphadenopathy, debility and non-responsiveness to antibiotic therapy were recorded. The animals responded to specific treatment against theileriosis. It is the first ever confirmed report of ovine theileriosis in Maharashtra state of India and hence reported.

scholarly journals Detection of Mycoplasma hyopneumoniae by ELISA and nested PCR from blood samples and nasal swabs from pigs in Slovakia

2012 ◽  
Vol 81 (4) ◽  
pp. 327-331 ◽  
Author(s):  
Marián Prokeš ◽  
Dagmar Zendulková ◽  
Kateřina Rosenbergová ◽  
František Treml ◽  
Anna Ondrejková ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Clinical Symptoms ◽  
Venous Blood ◽  
Clinical Signs ◽  
Mycoplasma Hyopneumoniae ◽  
Blood Samples ◽  
Paired Samples ◽  
Nasal Swabs ◽  
First Time ◽  
Primary Agent

The aim of our study was to map the situation of swine mycoplasmoses on four farms in the region of Eastern Slovakia. The primary agent of Enzootic pneumonia of swine isMycoplasma hyopneumoniae. After reviewing the health status of conventional herds and evaluation of clinical symptoms, paired samples of nasal swabs and venous blood samples were collected from 38 pigs with clinical signs of respiratory disease. Nasal swab samples were tested by nested PCR, while blood samples were used to detect antibodies againstM. hyopneumoniaeby blockingELISA. The presence ofM. hyopneumoniaewas confirmed by nested PCR in four pigs (10.5%) and by blocking ELISA in 16 pigs (42.1%) of all four farms. This work presents for the first time comparison of different methods to diagnoseM. hyopneumoniaeinfection on pig farms in Eastern Slovakia.

scholarly journals An outbreak of bluetongue virus serotype 9 in Southern Croatia

2011 ◽  
Vol 80 (4) ◽  
pp. 331-336 ◽  
Author(s):  
Eddy Listeš ◽  
Sanja Bosnić ◽  
Miroslav Benić ◽  
Josip Madić ◽  
Željko Cvetnić ◽  
...  
Keyword(s):  
Bluetongue Virus ◽  
Neutralization Test ◽  
Light Trap ◽  
Clinical Signs ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Virus Serotype ◽  
Serum Neutralization Test ◽  
First Time ◽  
Obsoletus Complex

The aim of this study was to provide a description of the first epidemic of bluetongue and the first survey on midges of the genus Culicoides in Croatia. Clinical signs were firstly observed on November 2001 in sheep in Konavle, Dubrovnik – Neretva County. During this epizootic the overall sheep morbidity and mortality were 5.2% (95% confidence interval (c.i.), 4.1-6.6%) and 2.29% (95% c.i., 1.6-3.3%), respectively. After the outbreak, 3,318 serum samples of ruminants from 53 villages of the Dubrovnik – Neretva County were examined for bluetongue virus (BTV) antibodies by competitive enzyme-linked immunosorbent assay (cELISA). In forty nine (92.45%, 95% c.i., 82.11-96.92%) of the 53 villages, animals with antibodies against bluetongue virus were found. In particular, a total of 178 cattle (49.86%, 95% c.i., 44.7-55.0%), 174 sheep (13.72%, 95% c.i., 11.9-15.7%) and 270 goats (15.95%, 95% c.i., 14.3-17.8%) were seropositive. Antibodies to bluetongue virus serotype 9 were detected in 212 positive sera by serum neutralization test. The percentage of positive animals decreased (P > 0.05) from the east to the west suggesting a possible east westward spreading of BTV infection. Fourteen light-trap midge collections from seven different sites were examined. Of the 4872 Culicoides spp. collected, 4,492 (92%, 95% c.i., 91.4-92.9%) of them belonged to the species of Obsoletus complex. This study showed for the first time that a pathogenic strain of BTV-9, probably from Montenegro, entered Croatia causing disease and death in local sheep and that C. obsoletus and C. scoticus were likely the major vectors of infection.

Correlation of avian bornavirus-specific antibodies and viral ribonucleic acid shedding with neurological signs and feather-damaging behaviour in psittacine birds

2019 ◽  
Vol 184 (15) ◽  
pp. 476-476 ◽  
Author(s):  
Alexandra Fluck ◽  
Dirk Enderlein ◽  
Anne Piepenbring ◽  
Ursula Heffels-Redmann ◽  
Sybille Herzog ◽  
...  
Keyword(s):  
Clinical Signs ◽  
Control Group ◽  
Clear Correlation ◽  
Serum Samples ◽  
Viral Shedding ◽  
Neurological Signs ◽  
Antibody Titres ◽  
Causative Agents ◽  
First Time ◽  
Proventricular Dilatation Disease

Parrot bornaviruses (PaBV) are the causative agents of proventricular dilatation disease in psittacine birds, but have also been linked to other clinical signs, including behavioural disorders and neurological signs. The aim of this study was to correlate PaBV infection in birds showing feather-damaging behaviour or neurological signs for which no other cause of disease could be identified. Psittacine birds presented to a private practice were divided into three groups: birds with neurological signs (n=28), birds showing feather-damaging behaviour (n=42) and birds presented for routine examinations (n=56). Swabs of crop and cloaca were collected and investigated for the presence of PaBV-RNA using real time RT-PCR. Additionally, serum samples were taken and examined for the presence of anti-PaBV antibodies by immunofluorescence test. PaBV infection was detected in one of the test systems in 40.5 per cent of all birds (n=126) investigated. In the clinically healthy birds (n=56), 19.6 per cent of the birds were positive in at least one of the PaBV tests, compared with 52.38 per cent of the feather-damaging (n=42) and 64.28 per cent of the neurologically diseased birds (n=28). Interestingly, the anti-PaBV antibody titres in birds with neurological signs were highest up to 1:20 480. High antibody titres (up to 1:5120) were also found in the feather-damaging group, whereas the birds of the control group, if PaBV positive, had only very low titres. Similarly, the highest viral load was found in the group of the neurologically diseased birds, followed by feather-damaging birds, whereas PaBV-positive birds in the control group demonstrated only low viral RNA shedding. A clear correlation between severity of clinical signs, amount of viral shedding and high levels of antibody titres was observed for most of the neurologically diseased birds and also for few birds with feather-damaging behaviour. For the first time, these results clearly indicate a correlation between PaBV infection and neurological signs in birds without gastrointestinal signs presented to the veterinarian in practice. It also may demonstrate a possible correlation with feather-damaging behaviour and anti-PaBV antibody presence. The antibody titre seems to represent a diagnostic tool to correlate clinical signs to PaBV as a cause.

scholarly journals Enzootic nasal adenocarcinoma in small ruminants in Jordan

2020 ◽  
Vol 51 (2) ◽  
pp. 219-223
Author(s):  
Wael Hananeh ◽  
Myassar El-Ekish ◽  
Rami Mukbel ◽  
Zuhair Bani Ismail ◽  
Mousa Daradka
Keyword(s):  
Histopathological Examination ◽  
Clinical Signs ◽  
Small Ruminants ◽  
Radiographic Examination ◽  
Low Grade ◽  
Cytological Examination ◽  
Awassi Sheep ◽  
Nasal Adenocarcinoma ◽  
Nasal Passages ◽  
First Time

Enzootic nasal adenocarcinoma (ENA) is a viral-associated tumor of the secretory epithelium of the nasal passages of small ruminants. The tumor is locally invasive but not metastatic. Although disease prevalence (up to 15%) and geographic distribution have been reported in the literature throughout most of the world (except Australia and New Zealand), the disease has until now remained unreported in Jordan. In this report, we describe the occurrence, clinical signs, radiographic signs and pathological features of the disease in a series of cases in Awassi sheep (3 cases) and Damascus breed goats (2 cases) for the first time in Jordan. Overall, the affected animals were adults (3-5 years of age), all females, with variable degrees of facial and nasal deformities resulting in difficulty in respiration. Radiographic examination of the skull revealed soft tissue density mass originating in the nasal turbinate and invading the surrounding nasal structures. Cytological examination of fine needle aspirate indicated low-grade carcinoma. All animals were humanely euthanized and the nasal passages were explored. Variable size masses were removed and histopathological examination confirmed the diagnosis of enzootic nasal adenocarcinoma.

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