Prevalence of <i>Pasteurella multocida</i> and other respiratory pathogens in the nasal tract of Scottish calves

Hotchkiss, Emily; Dagleish, Mark P.; Willoughby, Kim; McKendrick, Iain J.; Finlayson, J.; Zadoks, Ruth N.; Newsome, E.; Brülisauer, Franz; Gunn, George J.; Hodgson, John

doi:10.1136/vr.c4827

Cited by 37 publications

(36 citation statements)

References 21 publications

(18 reference statements)

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“…All methods were used as per the manufacturer's instructions. We have previously demonstrated that the QIAamp Viral RNA kit co-purifies BoHV-1 DNA from low cellularity samples [29] and, as BoHV-1 produces viral RNA during replication in tissues, for simplicity of clinical sample preparation we did not explicitly extract DNA for BoHV-1 detection. Phosphate buffered saline (PBS) was used as an extraction control for all extraction methods.…”

Section: Methodsmentioning

confidence: 99%

One-step multiplex real time RT-PCR for the detection of bovine respiratory syncytial virus, bovine herpesvirus 1 and bovine parainfluenza virus 3

et al. 2012

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BackgroundDetection of respiratory viruses in veterinary species has traditionally relied on virus detection by isolation or immunofluorescence and/or detection of circulating antibody using ELISA or serum neutralising antibody tests. Multiplex real time PCR is increasingly used to diagnose respiratory viruses in humans and has proved to be superior to traditional methods. Bovine respiratory disease (BRD) is one of the most common causes of morbidity and mortality in housed cattle and virus infections can play a major role. We describe here a one step multiplex reverse transcriptase quantitative polymerase chain reaction (mRT-qPCR) to detect the viruses commonly implicated in BRD.ResultsA mRT-qPCR assay was developed and optimised for the simultaneous detection of bovine respiratory syncytial virus (BRSV), bovine herpes virus type 1 (BoHV-1) and bovine parainfluenza virus type 3 (BPI3 i & ii) nucleic acids in clinical samples from cattle. The assay targets the highly conserved glycoprotein B gene of BoHV-1, nucleocapsid gene of BRSV and nucleoprotein gene of BPI3. This mRT-qPCR assay was assessed for sensitivity, specificity and repeatability using in vitro transcribed RNA and recent field isolates. For clinical validation, 541 samples from clinically affected animals were tested and mRT-qPCR result compared to those obtained by conventional testing using virus isolation (VI) and/or indirect fluorescent antibody test (IFAT).ConclusionsThe mRT-qPCR assay was rapid, highly repeatable, specific and had a sensitivity of 97% in detecting 102 copies of BRSV, BoHV-1 and BPI3 i & ii. This is the first mRT-qPCR developed to detect the three primary viral agents of BRD and the first multiplex designed using locked nucleic acid (LNA), minor groove binding (MGB) and TaqMan probes in one reaction mix. This test was more sensitive than both VI and IFAT and can replace the aforesaid methods for virus detection during outbreaks of BRD.

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Section: Methodsmentioning

confidence: 99%

One-step multiplex real time RT-PCR for the detection of bovine respiratory syncytial virus, bovine herpesvirus 1 and bovine parainfluenza virus 3

et al. 2012

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“…Likewise, a high prevalence of BRSV infection has been observed in Belgium [42]. Circulation of the virus in Scotland has also been reported, although conclusive figures regarding BRSV prevalence are not available [43]. Furthermore, the presence of antibodies in bulk tank milk samples has been reported in herds from England [44].…”

Section: Epidemiology Of Brsvmentioning

confidence: 99%

Epidemiology, Molecular Epidemiology and Evolution of Bovine Respiratory Syncytial Virus

Sarmiento-Silva

Nakamura-López²,

Vaughan

2012

Viruses

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The bovine respiratory syncytial virus (BRSV) is an enveloped, negative sense, single-stranded RNA virus belonging to the pneumovirus genus within the family Paramyxoviridae. BRSV has been recognized as a major cause of respiratory disease in young calves since the early 1970s. The analysis of BRSV infection was originally hampered by its characteristic lability and poor growth in vitro. However, the advent of numerous immunological and molecular methods has facilitated the study of BRSV enormously. The knowledge gained from these studies has also provided the opportunity to develop safe, stable, attenuated virus vaccine candidates. Nonetheless, many aspects of the epidemiology, molecular epidemiology and evolution of the virus are still not fully understood. The natural course of infection is rather complex and further complicates diagnosis, treatment and the implementation of preventive measures aimed to control the disease. Therefore, understanding the mechanisms by which BRSV is able to establish infection is needed to prevent viral and disease spread. This review discusses important information regarding the epidemiology and molecular epidemiology of BRSV worldwide, and it highlights the importance of viral evolution in virus transmission.

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“…It has been shown to cause respiratory disease in many animals, including cattle [2], sheep [3] and pigs [4,5] although it is also found in the respiratory tract of apparently healthy animals [6]. The organism also causes haemorrhagic septicaemia (HS) in bovids, mainly in South and Southeast Asia and sub-Saharan Africa [7].…”

Section: Introductionmentioning

confidence: 99%

Multilocus sequence typing of a global collection of Pasteurella multocida isolates from cattle and other host species demonstrates niche association

et al. 2011

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BackgroundPasteurella multocida causes disease in many host species throughout the world. In bovids, it contributes to bovine respiratory disease (BRD) and causes haemorrhagic septicaemia (HS). Previous studies have suggested that BRD-associated P. multocida isolates are of limited diversity. A multilocus sequence typing (MLST) scheme for P. multocida was used to determine whether the low levels of diversity reported are due to the limited discriminatory power of the typing method used, restricted sample selection or true niche association. Bovine respiratory isolates of P. multocida (n = 133) from the UK, the USA and France, collected between 1984 and 2008 from both healthy and clinically affected animals, were typed using MLST. Isolates of P. multocida from cases of HS, isolates from other host species and data from the MLST database were used as comparison.ResultsBovine respiratory isolates were found to be clonal (ISA 0.45) with 105/128 belonging to clonal complex 13 (CC13). HS isolates were not related to bovine respiratory isolates. Of the host species studied, the majority had their own unique sequence types (STs), with few STs being shared across host species, although there was some cross over between porcine and bovine respiratory isolates. Avian, ovine and porcine isolates showed greater levels of diversity compared to cattle respiratory isolates, despite more limited geographic origins.ConclusionsThe homogeneity of STs of bovine respiratory P. multocida observed, and the differences between these and P. multocida subpopulations from bovine non-respiratory isolates and non-bovine hosts may indicate niche association.

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Prevalence of Pasteurella multocida and other respiratory pathogens in the nasal tract of Scottish calves

Cited by 37 publications

References 21 publications

One-step multiplex real time RT-PCR for the detection of bovine respiratory syncytial virus, bovine herpesvirus 1 and bovine parainfluenza virus 3

One-step multiplex real time RT-PCR for the detection of bovine respiratory syncytial virus, bovine herpesvirus 1 and bovine parainfluenza virus 3

Epidemiology, Molecular Epidemiology and Evolution of Bovine Respiratory Syncytial Virus

Multilocus sequence typing of a global collection of Pasteurella multocida isolates from cattle and other host species demonstrates niche association

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