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Menangle and Nipah virus infections of pigs

Kirkland PD, Daniels PW, Nor MN, Love RJ, Philbey AW, Ross AD

Vet. Clin. North Am. Food Anim. Pract., 2002 Nov;18(3):557-71, ix.
PMID: 12442583

Viruses belonging to the family Paramyxoviridae generally have not been recognized as a significant cause of disease in pigs until recently. Between 1997 and 1999, there were large outbreaks of disease in pigs in Australia and Malaysia due to infection with viruses that have been shown to be new members of the Paramyxoviridae family. This article reviews current knowledge of Menangle and Nipah virus infections in pigs, the only major species of domestic animals to experience serious disease after infection with these viruses.

Matched MeSH terms: Swine Diseases/diagnosis
Fulltext Isolation of Mycoplasma hyosynoviae from pneumonic lung of swine

Dahlia H, Tan LJ, Zarrahimah Z, Maria J

Trop Biomed, 2009 Dec;26(3):341-5.
PMID: 20237449 MyJurnal

The isolation of Mycoplasma hyosynoviae from a piglet with severe pneumonia is described. This is the first report of M. hyosynoviae isolation in the country. The lung sample where the isolation was made was severely consolidated, suppurative and pleurisy. The pathogenicity of the M. hyosynoviae isolated has yet to be determined.

Matched MeSH terms: Swine Diseases/diagnosis
First molecular detection of porcine bocavirus in Malaysia

Mohan Jacob D, Lee CY, Arshad SS, Selvarajah GT, Bande F, Ong BL, et al.

Trop Anim Health Prod, 2018 Apr;50(4):733-739.
PMID: 29243138 DOI: 10.1007/s11250-017-1489-z

Several strains of porcine bocaviruses have been reported worldwide since their first detection in Sweden in 2009. Subsequently, the virus has been reported to be associated with gastrointestinal and respiratory signs in weaner and grower pigs. Although Malaysia is host to a self-sufficient swine livestock industry, there is no study that describes porcine bocavirus in the country. This report is the first to describe porcine bocavirus (PBoV) in Malaysian swine herds. PBoV was identified in various tissues from sick and runt pigs using the conventional PCR method with primers targeting conserved regions encoding for the nonstructural protein (NS1) gene. Out of 103 samples tested from 17 pigs, 32 samples from 15 pigs were positive for porcine bocavirus. In addition, a higher detection rate was identified from mesenteric lymph nodes (52.9%), followed by tonsil (37.0%), and lungs (33.3%). Pairwise comparison and phylogenetic analyses based on a 658-bp fragment of NS1 gene revealed that the Malaysian PBoV strains are highly similar to PBoV3 isolated in Minnesota, USA. The presence of porcine bocavirus in Malaysia and their phylogenetic bond was marked for the first time by this study. Further studies will establish the molecular epidemiology of PBoV in Malaysia and clarify pathogenicity of the local isolates.

Matched MeSH terms: Swine Diseases/diagnosis
Testing Eurasian wild boar piglets for serum antibodies against Mycobacterium bovis

Che' Amat A, González-Barrio D, Ortiz JA, Díez-Delgado I, Boadella M, Barasona JA, et al.

Prev Vet Med, 2015 Sep 1;121(1-2):93-8.
PMID: 26051843 DOI: 10.1016/j.prevetmed.2015.05.011

Animal tuberculosis (TB) caused by infection with Mycobacterium bovis and closely related members of the M. tuberculosis complex (MTC), is often reported in the Eurasian wild boar (Sus scrofa). Tests detecting antibodies against MTC antigens are valuable tools for TB monitoring and control in suids. However, only limited knowledge exists on serology test performance in 2-6 month-old piglets. In this age-class, recent infections might cause lower antibody levels and lower test sensitivity. We examined 126 wild boar piglets from a TB-endemic site using 6 antibody detection tests in order to assess test performance. Bacterial culture (n=53) yielded a M. bovis infection prevalence of 33.9%, while serum antibody prevalence estimated by different tests ranged from 19% to 38%, reaching sensitivities between 15.4% and 46.2% for plate ELISAs and between 61.5% and 69.2% for rapid immunochromatographic tests based on dual path platform (DPP) technology. The Cohen kappa coefficient of agreement between DPP WTB (Wildlife TB) assay and culture results was moderate (0.45) and all other serological tests used had poor to fair agreements. This survey revealed the ability of several tests for detecting serum antibodies against the MTC antigens in 2-6 month-old naturally infected wild boar piglets. The best performance was demonstrated for DPP tests. The results confirmed our initial hypothesis of a lower sensitivity of serology for detecting M. bovis-infected piglets, as compared to older wild boar. Certain tests, notably the rapid animal-side tests, can contribute to TB control strategies by enabling the setup of test and cull schemes or improving pre-movement testing. However, sub-optimal test performance in piglets as compared to that in older wild boar should be taken into account.

Matched MeSH terms: Swine Diseases/diagnosis*
Purification and characterization of Nipah virus nucleocapsid protein produced in insect cells

Eshaghi M, Tan WS, Ong ST, Yusoff K

J Clin Microbiol, 2005 Jul;43(7):3172-7.
PMID: 16000431

The nucleocapsid (N) protein of Nipah virus (NiV) is a major constituent of the viral proteins which play a role in encapsidation, regulating the transcription and replication of the viral genome. To investigate the use of a fusion system to aid the purification of the recombinant N protein for structural studies and potential use as a diagnostic reagent, the NiV N gene was cloned into the pFastBacHT vector and the His-tagged fusion protein was expressed in Sf9 insect cells by recombinant baculovirus. Western blot analysis of the recombinant fusion protein with anti-NiV antibodies produced a band of approximately 62 kDa. A time course study showed that the highest level of expression was achieved after 3 days of incubation. Electron microscopic analysis of the NiV recombinant N fusion protein purified on a nickel-nitrilotriacetic acid resin column revealed different types of structures, including spherical, ring-like, and herringbone-like particles. The light-scattering measurements of the recombinant N protein also confirmed the polydispersity of the sample with hyrdrodynamic radii of small and large types. The optical density spectra of the purified recombinant fusion protein revealed a high A(260)/A(280) ratio, indicating the presence of nucleic acids. Western blotting and enzyme-linked immunosorbent assay results showed that the recombinant N protein exhibited the antigenic sites and conformation necessary for specific antigen-antibody recognition.

Matched MeSH terms: Swine Diseases/diagnosis
Current status of diagnostic methods for henipavirus

Tamin A, Rota PA

Dev Biol (Basel), 2013;135:139-45.
PMID: 23689891 DOI: 10.1159/000189236

Hendra virus (HeV) and Nipah virus (NiV) are the causative agents of emerging transboundary animal disease in pigs and horses. They also cause fatal disease in humans. NiV has a case fatality rate of 40 - 100%. In the initial NiV outbreak in Malaysia in 1999, about 1.1 million pigs had to be culled. The economic impact was estimated to be approximately US$450 million. Worldwide, HeV has caused more than 60 deaths in horses with 7 human cases and 4 deaths. Since the initial outbreak, HeV spillovers from Pteropus bats to horses and humans continue. This article presents a brief review on the currently available diagnostic methods for henipavirus infections, including advances achieved since the initial outbreak, and a gap analysis of areas needing improvement.

Matched MeSH terms: Swine Diseases/diagnosis
Fulltext A RT-PCR assay for the detection of coronaviruses from four genera

Xiu L, Binder RA, Alarja NA, Kochek K, Coleman KK, Than ST, et al.

J Clin Virol, 2020 07;128:104391.
PMID: 32403008 DOI: 10.1016/j.jcv.2020.104391

BACKGROUND: During the past two decades, three novel coronaviruses (CoVs) have emerged to cause international human epidemics with severe morbidity. CoVs have also emerged to cause severe epidemics in animals. A better understanding of the natural hosts and genetic diversity of CoVs are needed to help mitigate these threats.
OBJECTIVE: To design and evaluate a molecular diagnostic tool for detection and identification of all currently recognized and potentially future emergent CoVs from the Orthocoronavirinae subfamily.
STUDY DESIGN AND RESULTS: We designed a semi-nested, reverse transcription RT-PCR assay based upon 38 published genome sequences of human and animal CoVs. We evaluated this assay with 14 human and animal CoVs and 11 other non-CoV respiratory viruses. Through sequencing the assay's target amplicon, the assay correctly identified each of the CoVs; no cross-reactivity with 11 common respiratory viruses was observed. The limits of detection ranged from 4 to 4 × 102 copies/reaction, depending on the CoV species tested. To assess the assay's clinical performance, we tested a large panel of previously studied specimens: 192 human respiratory specimens from pneumonia patients, 5 clinical specimens from COVID-19 patients, 81 poultry oral secretion specimens, 109 pig slurry specimens, and 31 aerosol samples from a live bird market. The amplicons of all RT-PCR-positive samples were confirmed by Sanger sequencing. Our assay performed well with all tested specimens across all sample types.
CONCLUSIONS: This assay can be used for detection and identification of all previously recognized CoVs, including SARS-CoV-2, and potentially any emergent CoVs in the Orthocoronavirinae subfamily.

Matched MeSH terms: Swine Diseases/diagnosis*
Fulltext A quantitative reverse transcription-polymerase chain reaction for detection of Getah virus

Sam SS, Teoh BT, Chee CM, Mohamed-Romai-Noor NA, Abd-Jamil J, Loong SK, et al.

Sci Rep, 2018 12 05;8(1):17632.
PMID: 30518924 DOI: 10.1038/s41598-018-36043-6

Getah virus (GETV), a mosquito-borne alphavirus, is an emerging animal pathogen causing outbreaks among racehorses and pigs. Early detection of the GETV infection is essential for timely implementation of disease prevention and control interventions. Thus, a rapid and accurate nucleic acid detection method for GETV is highly needed. Here, two TaqMan minor groove binding (MGB) probe-based quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assays were developed. The qRT-PCR primers and TaqMan MGB probe were designed based on the conserved region of nsP1 and nsP2 genes of 23 GETV genome sequences retrieved from GenBank. Only the qRT-PCR assay using nsP2-specific primers and probe detected all two Malaysia GETV strains (MM2021 and B254) without cross-reacting with other closely related arboviruses. The qRT-PCR assay detected as few as 10 copies of GETV RNA, but its detection limit at the 95% probability level was 63.25 GETV genome copies (probit analysis, P ≤ 0.05). Further validation of the qRT-PCR assay using 16 spiked simulated clinical specimens showed 100% for both sensitivity and specificity. In conclusion, the qRT-PCR assay developed in this study is useful for rapid, sensitive and specific detection and quantification of GETV.

Matched MeSH terms: Swine Diseases/diagnosis
Production of the matrix protein of Nipah virus in Escherichia coli: virus-like particles and possible application for diagnosis

Subramanian SK, Tey BT, Hamid M, Tan WS

J Virol Methods, 2009 Dec;162(1-2):179-83.
PMID: 19666056 DOI: 10.1016/j.jviromet.2009.07.034

The broad species tropism of Nipah virus (NiV) coupled with its high pathogenicity demand a rapid search for a new biomarker candidate for diagnosis. The matrix (M) protein was expressed in Escherichia coli and purified using a Ni-NTA affinity column chromatography and sucrose density gradient centrifugation. The recombinant M protein with the molecular mass (Mr) of about 43 kDa was detected by anti-NiV serum and anti-myc antibody. About 50% of the M protein was found to be soluble and localized in cytoplasm when the cells were grown at 30 degrees C. Electron microscopic analysis showed that the purified M protein assembled into spherical particles of different sizes with diameters ranging from 20 to 50 nm. The purified M protein showed significant reactivity with the swine sera collected during the NiV outbreak, demonstrating its potential as a diagnostic reagent.

Matched MeSH terms: Swine Diseases/diagnosis*
A review of Nipah and Hendra viruses with an historical aside

Ksiazek TG, Rota PA, Rollin PE

Virus Res, 2011 Dec;162(1-2):173-83.
PMID: 21963678 DOI: 10.1016/j.virusres.2011.09.026

The emergence of Hendra and Nipah viruses in the 1990s has been followed by the further emergence of these viruses in the tropical Old World. The history and current knowledge of the disease, the viruses and their epidemiology is reviewed in this article. A historical aside summarizes the role that Dr. Brian W.J. Mahy played at critical junctures in the early stories of these viruses.

Matched MeSH terms: Swine Diseases/diagnosis