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

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.

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Serological, Hematologic, and PCR Studies of Cattle in an Area of Switzerland in Which Tick-Borne Fever (Caused by Ehrlichia phagocytophila) Is Endemic

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.5.3.325-327.1998 ◽

1998 ◽

Vol 5 (3) ◽

pp. 325-327 ◽

Cited By ~ 9

Author(s):

Nicola Pusterla ◽

Jeannine Berger Pusterla ◽

Ueli Braun ◽

Hans Lutz

Keyword(s):

Seasonal Variations ◽

Nested Pcr ◽

Molecular Detection ◽

Clinical Signs ◽

Pcr Amplification ◽

Blood Samples ◽

Ehrlichia Phagocytophila ◽

Blood Smears ◽

Antibody Titers ◽

First Time

ABSTRACT The purpose of this study was to examine the seasonal variations in seroprevalence to Ehrlichia phagocytophila in cattle pastured during the summer months in an area where tick-borne fever is endemic. The study was performed during a 1-year period from April 1996 to March 1997 and involved 34 cows, 22 pregnant heifers, and 14 calves. Blood samples, collected from all 70 cattle once a month, were used to determine serum immunoglobulin G titers by indirect immunofluorescence. In addition, blood smears were examined for Ehrlichiaorganisms, and PCR amplification was performed for the molecular detection of E. phagocytophila. Prior to the pasture period, the seroprevalence was 16%. Two weeks after the start of pasturing, it was 43%, after which it progressively increased and reached a maximum of 63% in September. The seroprevalence progressively decreased after the end of pasturing to a low of 23%. The variation in antibody titers was similar to that of seroprevalence.E. phagocytophila organisms were detected in blood smears of 7 animals and by nested PCR in 12. Only four cows, which were on the pastures of endemicity for the first time, had clinical signs of ehrlichiosis. This study demonstrated marked seasonal variations in seroprevalence and in serum titers of antibody to E. phagocytophila in cattle. The incidence of clinical signs of ehrlichiosis was increased in cattle grazing on the pastures of endemicity for the first time.

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Chlamydophila psittaci genotype E/B transmission from African grey parrots to humans

Journal of Medical Microbiology ◽

10.1099/jmm.0.47157-0 ◽

2007 ◽

Vol 56 (8) ◽

pp. 1097-1100 ◽

Cited By ~ 26

Author(s):

Taher Harkinezhad ◽

Kristel Verminnen ◽

Caroline Van Droogenbroeck ◽

Daisy Vanrompay

Keyword(s):

Nested Pcr ◽

Clinical Symptoms ◽

Clinical Signs ◽

Pharyngeal Swab ◽

Loss Of Weight ◽

Nasal Swabs ◽

Chlamydophila Psittaci ◽

Highly Virulent ◽

Genotype A ◽

Genotype E

Thirty-six birds from a parrot relief and breeding centre, as well as the manager, were examined for the presence of Chlamydophila psittaci. In the relief unit, 5 of 20 African grey parrots showed depression, ruffled feathers, loss of weight and mild dyspnoea. The birds received no antibiotic treatment. Birds of the breeding unit, 14 blue and gold macaws and 2 green-winged macaws, were healthy. They received doxycycline at the start of each breeding season. The manager complained of shortness of breath but took no medication. Using a nested PCR enzyme immunoassay (EIA), Cp. psittaci was detected in the faeces of all five sick birds, as well as in a nasal and pharyngeal swab from the manager. The veterinarian and her assistant became infected while sampling the parrots, as pharyngeal and nasal swabs from both were positive by nested PCR/EIA after visiting the parrot relief and breeding centre, but they showed no clinical signs of infection. Bacteria could be isolated from three of five nested PCR/EIA-positive birds, the manager and the veterinarian, but not from the veterinary assistant. Using an ompA genotype-specific real-time PCR, Cp. psittaci genotype E/B was identified as the transmitted strain. All breeding birds tested negative for Cp. psittaci. This is believed to be the first report on Cp. psittaci genotype E/B transmission from parrots to humans. In contradiction to genotype A strains, which are thought to be highly virulent to both birds and men, the currently described genotype E/B strain apparently caused no severe clinical symptoms in either parrots or humans.

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First Report on the Detection and Molecular Characterization of Bovine Herpesvirus 1 from a Clinical case of Infectious Bovine Rhinotracheitis in Pakistan

Pakistan Veterinary Journal ◽

10.29261/pakvetj/2020.084 ◽

2021 ◽

Vol 41 (01) ◽

pp. 160-162

Author(s):

Aayesha Riaz

Keyword(s):

Molecular Characterization ◽

Nested Pcr ◽

Bovine Herpesvirus ◽

Nasal Discharge ◽

Infectious Bovine Rhinotracheitis ◽

Bovine Herpesvirus 1 ◽

Nasal Swabs ◽

Herpesvirus 1 ◽

First Time

Bovine herpesvirus 1 (BHV-1) is a noteworthy reason for many Cattle/Buffalo diseases. Infectious bovine rhinotracheitis (IBR) is one of the diseases which are caused by the BHV-1. In the present study a cow which was suspected of IBR was examined. The animal was suffering from fever and respiratory distress along with rhinitis (red nose), in appetence, and dyspnea. The nasal mucosa and muzzle were distinctly inflamed, with nasal discharge. DNA from blood samples and nasal swabs were subjected to nested PCR using glycoprotein B gene (gB) Primers. The samples were found positive for BHV-1 gB gene. Sequencing and phylogenetic analysis revealed close similarities with other BHV-1 gB gene sequences. The accession numbers assigned to this pioneer sequences in GenBank are MT449510 (BHV-1-Pak 1) and MT449511 (BHV-1-Pak 2). In this study, we reported for the first time the detection of DNA of BHV-1 through nested PCR assay and molecular characterization of BHV-1 gB gene in Pakistan. This study will be useful in further diagnoses of BHV-1 in Pakistan and in development of BHV-1 vaccine to reduce economical losses due to IBR

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Molecular detection of bovine hemotrophic mycoplasmas in Mexico

Revista del Centro de Investigación de la Universidad la Salle ◽

10.26457/recein.v13i52.2183 ◽

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.

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Hot flush values of gonadotropins and estradiol in menopause

Srpski arhiv za celokupno lekarstvo ◽

10.2298/sarh160303012a ◽

2018 ◽

Vol 146 (7-8) ◽

pp. 403-406

Author(s):

Zorana Arizanovic ◽

Svetlana Vujovic ◽

Miomira Ivovic ◽

Milina Tancic-Gajic ◽

Ljiljana Marina ◽

...

Keyword(s):

Clinical Symptoms ◽

Venous Blood ◽

Hot Flashes ◽

Thyroid Stimulating Hormone ◽

Sex Hormone Binding Globulin ◽

Control Group ◽

Subjective Feeling ◽

Blood Samples ◽

Hot Flush ◽

Stimulating Hormone

Introduction/Objective. Hot flashes are one of the first clinical symptoms of menopause. The mechanism of hot flashes is still not fully understood. Changes in concentrations of the circulating follicle-stimulating hormone (FSH), luteinizing hormone (LH), estrogen, and other hormones can lead to thermoregulatory dysfunction. The aim of this study was to examine the association between dynamic changes in concentrations of sex hormones and the presence of vasomotor symptoms in menopausal women. Methods. The study involved 36 women divided into two groups: in the first group there were 24 women with hot flashes, BMI 26.16 ? 3.42 kg/m2; the control group comprised 12 women, BMI 26.82 ? 3.89 kg/m2. Data on the presence of hot flashes were based on medical history data. Venous blood samples were collected for the analyses of FSH, LH, prolactin, estradiol, progesterone, testosterone, sex hormone binding globulin, dehidroepiandrosteron sulfate, thyroid-stimulating hormone, and thyroxin. During the subjective feeling of hot flashes, three blood samples during the day and night were collected to determine the mean levels of FSH, LH, and estradiol in women with hot flashes. Results. Women with hot flashes had significantly higher prolactin (389.58 ? 123.69 mIU/L to 258.19 ? 122 mIU/L, p < 0.01) and dehydroepiandrosterone sulfate (3.60 ? 2.49 nmol/L vs. 1.88 ? 1.27 nmol/L, p < 0.05) levels, as well as lower mean values of FSH during hot flashes during the day (69.08 ? 28.84 IU/L vs. 107.18 ? 39.11 IU/L, p < 0.01) and night (60.72 ? 21.89 IU/L vs. 104.57 ? 38.06 IU/L, p < 0.01). Conclusion. .Women with hot flashes had significantly lower mean FSH levels during hot flashes during the day and night than the control group.

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Infection status of Mycoplasma hyopneumoniae in experimental pigs at a commercial farm

The Journal of Agriculture and Development ◽

10.52997/jad.4.03.2020 ◽

2020 ◽

Vol 19 (03) ◽

pp. 22-27

Author(s):

Huyen T. N. Bui

Keyword(s):

Oral Fluid ◽

Humoral Response ◽

Mycoplasma Hyopneumoniae ◽

Infection Status ◽

Blood Samples ◽

Maternal Immunity ◽

Nasal Swabs ◽

The One ◽

Very High ◽

Experimental Pigs

The objective of this study was to investigate the profiles of Mycoplasma hyopneumoniae (MH) infection at different ages of pig in a sow – finishing herd using serological and molecular methods. A total of 30 study piglets were born from non-vaccinated sows with MH. They were injected one-dose of inactivated MH vaccine at the 10th week. MH infection status was evaluated by using ELISA to detect MH antibodies from blood samples, and PCR to detect MH DNA in nasal swabs or oral fluid samples every other week from newborn to slaughter time. The results of this study showed that PCR positive proportions were low at 1st-2nd week (7-13%), then increased significantly during 5th -7th week (73-79%) and reduced at 8th week (33%); finally became negative after 13th week of age. This pattern corresponds to the one of antibody level. In particular, the level of maternal antibodies against MH was very high due to maternal immunity, then decreased gradually to negative at 7-8 weeks of age, and finally increased gradually from 13 weeks of age to all positive at 25 weeks of age. In conclusion, the result showed that in this herd, MH might invade pigs by the time of 5-7 weeks of age after maternal immunity disappears, and humoral response can overcome the infection at week 13. This should be noted to have appropriate strategies to control MH at the farm.

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Diagnosis of a Dicrocoelium dendriticum infection in New World Camelids: a case report

Veterinární Medicína ◽

10.17221/5856-vetmed ◽

2012 ◽

Vol 57 (No. 3) ◽

pp. 154-162 ◽

Cited By ~ 2

Author(s):

D. Klein ◽

H. Prosl ◽

D. Thaller ◽

M. Floeck

Keyword(s):

Bile Ducts ◽

New World ◽

Liver Enzymes ◽

Clinical Symptoms ◽

Clinical Signs ◽

Dicrocoelium Dendriticum ◽

History Of ◽

Liver Flukes ◽

The University ◽

First Time

Dicrocoelium dendriticum plays an important role in New World Camelids as infected animals may suffer from severe clinical symptoms even leading to death of the animals. Intra vitam diagnosis may be difficult as clinical signs are atypical and Dicrocoelium eggs are shed only intermittently in faeces. The aim of this paper is to present four clinical cases of dicrocoeliosis in lamas as well as three asymptomatic infected animals to support the veterinarian in practice to diagnose infections. Furthermore, it is the first time that ultrasonographic examinations are described in this context. All seven lamas had been admitted to the Clinic for Ruminants at the University for Veterinary Medicine in Vienna. None of the animals had a history of D. dendriticum infection. The ultrasonographic examination of the liver revealed in all diseased animals as well as in two asymptomatic lamas hyperechoic areas representing calcified bile ducts typical for an infection with liver flukes. These findings together with blood examination of liver enzymes and parasitological examination may lead to the intra vitam diagnosis of dicrocoeliosis in lamas and alpacas. With an early diagnosis, the therapy of Dicrocoelium spp. could become more effective and the number of animals rescued may be increased.  

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Mycoplasma hyopneumoniae as a causative agent of porcine enzootic pneumonia

Livestock ◽

10.12968/live.2021.26.3.136 ◽

2021 ◽

Vol 26 (3) ◽

pp. 136-143

Author(s):

Christina Gale ◽

Eduardo Velazquez ◽

Emma Pattison

Keyword(s):

Management Practices ◽

Clinical Symptoms ◽

Respiratory Health ◽

Swine Influenza Virus ◽

Clinical Signs ◽

Mycoplasma Hyopneumoniae ◽

Herd Health ◽

Enzootic Pneumonia ◽

Lung Lesions ◽

The Impact

Mycoplasma hyopneumoniae is a very important pathogen which causes enzootic pneumonia (EP) affecting pig herds worldwide. M. hyopneumoniae establishes itself in the respiratory system causing pathological changes, and resultant pneumonic lesions can be detected at slaughter via lung examination, appearing as red to pink lesions which resolve over time, leaving scar tissue on the lobes. Mycoplasma spp. infection can be complicated by coinfections with other pathogens such as swine influenza virus (SIV) and porcine circovirus (PCV2). These coinfections result in worsening of clinical symptoms and lung lesions, therefore increasing the impact on performance of the affected animals. Therefore, it is important that focus is placed on improving overall respiratory health and managing overall herd health, rather than just solely focusing on M. hyopneumoniae. Management practices should be implemented to control the disease, which commonly involve vaccination to reduce the impacts of the disease. Effective vaccination is important, and can be ensured by correct vaccine handling, storage and administration. The vaccine used can also play a part in the success of the vaccination programme so this should also be assessed, which can be done via lung examination and noting of clinical signs and performance parameters. When vaccination is successful, the level of lung lesions should be reduced, performance improved and consistency between batches, demonstrating that respiratory health is more stable.

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

Viruses ◽

10.3390/v12090983 ◽

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.

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Development and Evaluation of a Molecular Diagnostic Method for Rapid Detection of Histoplasma capsulatum var. farciminosum, the Causative Agent of Epizootic Lymphangitis, in Equine Clinical Samples

Journal of Clinical Microbiology ◽

10.1128/jcm.00896-16 ◽

2016 ◽

Vol 54 (12) ◽

pp. 2990-2999

Author(s):

C. E. Scantlebury ◽

G. L. Pinchbeck ◽

P. Loughnane ◽

N. Aklilu ◽

T. Ashine ◽

...

Keyword(s):

Causative Agent ◽

Nested Pcr ◽

Diagnostic Method ◽

Histoplasma Capsulatum ◽

Clinical Signs ◽

Molecular Diagnostic ◽

Clinical Samples ◽

Blood Samples ◽

Content Type ◽

Molecular Diagnostic Method

Histoplasma capsulatumvar.farciminosum, the causative agent of epizootic lymphangitis (EZL), is endemic in parts of Africa. Diagnosis based on clinical signs and microscopy lacks specificity and is a barrier to further understanding this neglected disease. Here, a nested PCR method targeting the internal transcribed spacer (ITS) region of the rRNA operon was validated for application to equine clinical samples. Twenty-nine horses with signs of EZL from different climatic regions of Ethiopia were clinically examined. Blood samples and aspirates of pus from cutaneous nodules were taken, along with blood from a further 20 horses with no cutaneous EZL lesions. Among the 29 horses with suspected cases of EZL,H. capsulatumvar.farciminosumwas confirmed by extraction of DNA from pus and blood samples from 25 and 17 horses, respectively. Positive PCR results were also obtained with heat-inactivated pus (24 horses) and blood (23 horses) spotted onto Whatman FTA cards. Two positive results were obtained among blood samples from 20 horses that did not exhibit clinical signs of EZL. These are the first reports of the direct detection ofH. capsulatumvar.farciminosumin equine blood and at high frequency among horses exhibiting cutaneous lesions. The nested PCR outperformed conventional microscopic diagnosis, as characteristic yeast cells could be observed only in 14 pus samples. The presence ofH. capsulatumvar.farciminosumDNA was confirmed by sequencing the cloned PCR products, and while alignment of the ITS amplicons showed very little sequence variation, there was preliminary single nucleotide polymorphism-based evidence for the existence of two subgroups ofH. capsulatumvar.farciminosum. This molecular diagnostic method now permits investigation of the epidemiology of EZL.

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