Roongroje Thanawongnuwech

Chulalongkorn University

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Sanipa Suradhat

Chulalongkorn University

Roongtham Kedkovid

Chulalongkorn University

Alongkorn Amonsin

Chulalongkorn University

Chaitawat Sirisereewan

Chulalongkorn University

Jirapat Arunorat

Chiang Mai University

Komkrich Teankum

Chulalongkorn University

Yonlayong Woonwong

Kasetsart University

Suphattra Jittimanee

Khon Kaen University

Sawang Kesdangsakonwut

Chulalongkorn University

Yong Poovorawan

Chulalongkorn University

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Chulalongkorn University

135

Iowa State University

Mahidol University

Chiang Mai University

Kasetsart University

United States Department of Agriculture

Agricultural Research Service

PTT Public Company Limited (Thailand)

University of Minnesota

Nong Lam University Ho Chi Minh City

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PULMONNARY INTRAVASCULAR MACROPHAGES (PIMs): WHAT DO WE KNOW ABOUT THEIR ROLE IN PRRSV INFECTION IN PIGS?

The Thai Journal of Veterinary Medicine (1999)

Roongroje Thanawongnuwech

Porcine pulmonary intravascular macrophages (PIMs) have been recently known for their role of pulmonary surveillance. In the past few years, we have been investigating the role of PIMs in porcine reproductive and respiratory syndrome virus (PRRSV) infection. PRRSV antigen has been demonstrated in PIMs both in vitro and in vivo. PRRSV-infected PIMs reveal viral particles that tend to accumulate in the smoothed-wall vesicles and yield a high virus titer. The PRRSV infection induces either apoptosis or cell lysis. The bactericidal activity of in vitro PRRSV infected PIMs is significantly decreased and the phagocytic activity is questionable as the pulmonary copper clearance, which measures the phagocytic activity of PIMs in vivo, is significantly decreased in PRRSV-infected pigs. This evidence supports the hypothesis that PRRSV - induced damage to PIMs results in increased susceptibility to bacteremic diseases. This result could explain the increase in the chronic bacterial respiratory diseases, septicemia, and mortality experienced in pigs on farms endemically-or epizootically-infected with highly virulent strains of PRRSV. The objective of this article is to summarize the current knowledge of the complex interaction between PRRSV and the PIMs.

10.56808/2985-1130.1775

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Comparative Efficacy of Chimeric Porcine Circovirus (PCV) Vaccines against Experimental Heterologous PCV2d Challenges

Veterinary Sciences (2023)

Pichanun Wongchanapai Panuwat Yamsakul Jirapat Arunorat Thunyamas Guntawang Tidaratt Sittisak

The objective of this study was to evaluate the efficacy of two multivalent commercial porcine circovirus (PCV) vaccines against heterologous PCV2d challenges. A total of 24 crossbred male pigs aged 26 days selected from a specific pathogen-free herd were randomly divided into four groups (six pigs per group) and assigned as follows: negative control (unvaccinated/sham-challenge), vaccinated with chimeric PCV1-2a vaccine (PCV1-2a/PCV2d-challenge), vaccinated with chimeric PCV1-2a-2b vaccine (PCV1-2a-2b/PCV2d-challenge) and positive control (unvaccinated/PCV2d-challenge). At 21 days after vaccination, the pigs were intranasally and intramuscularly inoculated with either sham or field isolates of PCV2d (PCV2d/149/TH/2020). After being challenged, blood samples were obtained weekly and analyzed for levels of PCV2d viremia, neutralizing antibodies, and IgG against PCV2. At 30 days post-challenge (DPC), the pigs were euthanized and then subjected to pathological evaluations and molecular analysis. The results indicated that pigs in the PCV1-2a-2b/PCV2d-challenge and the PCV1-2a/PCV2d-challenge groups possessed significantly greater levels of PCV2d-neutralizing antibody titer when compared with the positive control group. Moreover, pigs in the PCV1-2a-2b/PCV2d-challenge group exhibited a lower degree of severity in terms of gross lesion scores and lower levels of PCV2 viremia when compared with the positive control group. This study demonstrated that vaccinating pigs with either the PCV1-2a or PCV1-2a-2b chimeric vaccines elicits a potent immune response against PCV2d infection and reduces viremia after PCV2d inoculation in pigs.

10.3390/vetsci10020080

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An indirect enzyme-linked immunosorbent assay using a recombinant truncated capsid protein of Porcine circovirus-2

Journal of Veterinary Diagnostic Investigation (2012)

Suphattra Jittimanee Suparlark Nuntawan Na Ayudhya Roongtham Kedkovid Komkrich Teankum Sanipa Suradhat

Porcine circovirus-2 (PCV-2) serology is commonly used for PCV-2 herd status determination and optimal timing of PCV-2 vaccination programs. The objectives of the current study were to develop an in-house indirect enzyme-linked immunosorbent assay (ELISA) using a recombinant nuclear localization signal truncated capsid (rntCap) protein expressed in an Escherichia coli system and to determine the diagnostic performance of the developed rntCap indirect ELISA in comparison with immunoperoxidase monolayer assays (IPMAs). Based on a receiver operating characteristic (ROC) curve analysis of the rntCap indirect ELISA ( n = 90), an optimum cutoff optical density (OD) of 0.330 was determined, which resulted in diagnostic sensitivity, diagnostic specificity, and accuracy of 98.33%, 93.33%, and 96.67%, respectively. Average OD values of the positive ( n = 8) and negative sera ( n = 8) tested by either purified glutathione-S-transferase (GST) protein or the rntCap protein as the coating antigen revealed that the mean OD values tested by the rntCap indirect ELISA were significantly different from using GST alone ( P < 0.005). The correlation between the established rntCap indirect ELISA and the IPMA results demonstrated as the linear regression (Spearman correlation coefficient = 0.772, P < 0.005) indicated that the OD ratio obtained from the rntCap indirect ELISA could be used to predict the levels of the IPMA titers. More samples are needed for enhancing the diagnostic sensitivity, specificity and accuracy. In conclusion, the establishment of the rntCap indirect ELISA could be used as a serodiagnostic assay for large-scale detection of PCV-2 antibodies in swine and has the capability to be produced commercially for routine use in diagnostic laboratories.

10.1177/1040638712461251

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Citations (9)

Transdermal delivery of plasmid encoding truncated nucleocapsid protein enhanced PRRSV-specific immune responses

Vaccine (2015)

Sanipa Suradhat Piya Wongyanin Chaitawat Sirisereewan Teerawut Nedumpun Mongkol Lumyai

10.1016/j.vaccine.2015.12.043

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Development of Nonstructural Protein-Based Indirect ELISA to Identify Elephant Endotheliotropic Herpesvirus (EEHV) Infection in Asian Elephants (Elephas maximus)

Animals (2022)

Thunyamas Guntawang Tidaratt Sittisak Pallop Tankaew Chatchote Thitaram Varangkana Langkapin

Disease caused by elephant endotheliotropic herpesvirus (EEHV) infection is the most highly fatal hemorrhagic disease in Asian elephant calves worldwide. To date, adult elephants that have been infected with EEHV have predominantly displayed mild clinical signs, while they are believed to serve as EEHV shedders to other elephants. Hence, the diagnostic tools employed for monitoring EEHV-active infection are considered vitally important. In this study, partial EEHV-DNA polymerase (DNApol) nonstructural proteins (NSPs) were used to detect anti-EEHV antibodies through the use of an in-house indirect enzyme-linked immunosorbent assay (ELISA). The results were then compared to those obtained from a PCR test. In this study, a total of 175 serum samples were collected from Asian elephants living in elephant camps located in Chiang Mai and Lampang Provinces, Thailand. The elephants were aged between 2 and 80 years old. The overall percentages of positive samples by the PCR and EEHV-DNApol ELISA tests were 4% (21/175) and 12% (21/175), respectively. The ELISAs demonstrated values of 77.9% (95% posterior probability interval (PPI) = 52.5-95%) sensitivity and 87.7% (PPI = 82.5-91.9%) specificity, respectively. Accordingly, the sera obtained from the elephants exhibiting no clinical signs of EEHV infection, and those who were negative according to PCR tests, revealed a value of 14% seropositivity for EEHV-DNApol. Our results indicate that these asymptomatic, active EEHV-infected elephants could likely serve as a source of EEHV shedding within elephant herds. Consequently, the developed EEHV-DNApol NSPs-based ELISA test employed in the present study may be of use for routine monitoring and identification of EEHV shedders in elephant herds, and could be an alternative diagnostic tool for EEHV detection in Asian elephants.

Asian elephant

10.3390/ani12141747

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Citations (2)

Dual infection of a Thai isolate HP-PRRSV and the pdmH1N1 2009 SIV in weanling pigs

The Thai Journal of Veterinary Medicine (2019)

Nattaphong Sangpratum Roongtham Kedkovid Yonlayong Woonwong Chaitawat Sirisereewan Sawang Kesdangsakonwut

After the introduction of the pandemic H1N1 swine influenza virus (pdmH1N1 SIV) in Thai swine farms in 2009, a mild clinical outcome was observed and its role in porcine respiratory disease complex (PRDC) is questionable. A highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) has become endemic in Thai swine farms, recently. This study was conducted to determine the pathogenicity of the pdmH1N1 SIV when co-infected with a HP-PRRSV. Thirty-two weanling pigs from a free PRRSV, SIV, porcine circovirus type 2 (PCV-2) and Mycoplasma spp. commercial farm were randomly divided into 4 groups; Negative, PRRSV, the SIV and Co-infection groups. Pigs in the PRRSV and Co-infection groups were challenged intranasally with the Thai isolate HP-PRRSV at 0 day post HPPRRSV challenge (dphc) and pigs in the SIV and Co-infection groups were inoculated intratracheally with the pdmH1N1 SIV at 6 dphc. The 1st and 2nd necropsies were at 2 and 4 days post SIV challenge (dsc). A comparison of the lung lesion scores between the PRRSV and Co-infection groups at 2 dsc showed no statistical difference, but at 4 dsc, the lung lesion scores of the Co-infection group had higher average scores than those of the PRRSV group. Similarly, the SIV viral load in the lung of the Co-infection group a showed higher viral load at 4 dsc, compared to the only SIV-infected group. The results suggest that low pathogenic pdmH1N1 SIV could enhance the lung severity when co-infected with the HP-PRRSV leading to PRDC mimicking the field scenarios.

Weanling

10.56808/2985-1130.2976

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Genetic Characterization and Phylogenetic Analysis of Porcine Circovirus Type 2 in Thai Pigs with Porcine Circovirus Associated Diseases (PCVAD) during 2007-2010

The Thai Journal of Veterinary Medicine (2011)

Suphattra Jittimanee S. Na Roongtham Kedkovid Komkrich Teankum Roongroje Thanawongnuwech

Porcine circovirus type 2 (PCV2) was first recognized as a causative agent of post weaning multisystemicsyndrome or PMWS in Canada in 1991. Subsequently, it has been reported in almost all intensive pig productioncountries, and currently all associated diseases linked to PCV2 are included under the term porcine circovirusassociated diseases or PCVAD. PCV2 can be divided into 2 major genotypes referred to genotype 1 (PCV2b) andgenotype 2 (PCV2a). In Thailand, PMWS was first reported in 1998. However, only one isolate of Thai PCV2 from ourgroup has been submitted to GenBank (AY864814). Therefore, the objective of this study was to determine thegenetic characterizations of 12 ORF2 of PCV2 isolates from Thai pigs with PCVAD during 2007-2010. Twelve ThaiPCV2 sequences were analyzed together with 19 representative ORF2 sequences. Interestingly, all 12 recent ThaiPCV2 sequences belong to genotype 1 consisting of subgroup 1A/B (10/12, 83.33%) and 1C (2/12, 16.67%). Insummary, based on genetic characterization and phylogenetic analysis, a few specific substitution patterns wereobserved in amino acid positions and genotype 1, subgroup 1A/B was predominated in Thai pigs with PCVAD inthis study. More samples are needed to investigate the existence of genotype 2.

Circovirus

Classical swine fever

Genetic Analysis

10.56808/2985-1130.2292

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Citations (5)

Interleukin-10, Interleukin-12, and Interferon-γLevels in the Respiratory Tract FollowingMycoplasma hyopneumoniaeand PRRSV Infection in Pigs

Viral Immunology (2003)

Roongroje Thanawongnuwech Eileen L. Thacker

The cytokine profile associated with either a T helper 1 (Th1) or Th2 response in a porcine respiratory disease model was assessed by measuring IL-12, IL-10 and IFN-γ using RT-PCR and ELISA, respectively. IL-10, IL-12, and IFN-γ levels in pulmonary alveolar macrophages and bronchial lavage fluid were increased in pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV), Mycoplasma hyopneumoniae, or both pathogens. At 10 days post-infection (DPI), both IL-10 and IL-12 mRNA levels were increased in both groups infected with PRRSV. The IL-12 levels were increased in pigs infected with both pathogens and IFN-γ protein levels were increased in pigs infected with PRRSV alone and only numerically increased in the dual infection. At 28 DPI, IL-12 mRNA levels and IL-10 protein levels were increased in all infected groups. The mRNA level of IL-12 remained elevated in the group infected with both pathogens at 42 DPI. Production of IFN-γ did not appear to be closely correlated with elimination of virus from the respiratory tract. However, when the virus existed in the lung, the local IFN-γ production appeared to increase. Although IL-12 mRNA levels were significantly elevated in the pigs infected with both pathogens, the increased protein levels of IL-12 may compromise the immune system's ability to clear PRRSV from the lung. This could explain the prolonged presence of PRRSV, IFN-γ production and the increased pneumonia observed in the lungs of dual-infected pigs. The increased levels of cytokines associated with both Th1 and Th2 responses in the respiratory tract of pigs infected with PRRSV and M. hyopneumoniae provides valuable information on the pathogenesis of these diseases.

Mycoplasma hyopneumoniae

Respiratory tract

10.1089/088282403322396154

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Citations (67)

Comparison of Vaccination Regimens on Immune Responses Using PED Replicon Vaccine: A Field Trial in PED-Negative and PED-Positive Thai Swine Farms

Animals (2025)

Chaitawat Sirisereewan Thanh Che Nguyen Nanthiya Iampraphat Hongyao Lin Leonardo Ellerma

Porcine epidemic diarrhea virus (PEDV) is an economically important pathogen of swine, causing severe diarrhea in neonates with high morbidity and mortality. Vaccination is a key strategy for PEDV control, but optimizing regimens based on herd status is essential for improving immunity and protection. This study evaluated immune responses to different vaccination protocols using a PED replicon vaccine (PED-RP) in Thai swine farms with varying PED statuses. Two field trials were conducted: Experiment 1 (n = 120) on a PED-negative farm used a killed vaccine followed by PED-RP, while Experiment 2 (n = 42) on a PED-positive farm used a modified-live vaccine (MLV) followed by PED-RP. In Experiment 1, gilts boosted with PED-RP during gestation showed significantly higher colostrum IgA levels compared to those receiving two doses of the killed vaccine. In Experiment 2, priming with MLV followed by PED-RP resulted in increased neutralizing antibody titers and seropositivity, outperforming the MLV-only regimen after acclimatization. These findings suggest that using PED-RP as a booster can enhance serum and lactogenic immunity, providing a promising strategy for improving immune responses in both PED-negative and PED-positive herds. The study highlights the importance of tailored vaccination strategies to optimize herd immunity and protect piglets from PEDV.

Herd Immunity

Vaccine efficacy

10.3390/ani15020273

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Genetic characterization of 2008 reassortant influenza A virus (H5N1), Thailand

Virology Journal (2010)

Alongkorn Amonsin Jiradej Lapkuntod Kamol Suwannakarn Pravina Kitikoon Sanipa Suradhat

In January and November 2008, outbreaks of avian influenza have been reported in 4 provinces of Thailand. Eight Influenza A H5N1 viruses were recovered from these 2008 AI outbreaks and comprehensively characterized and analyzed for nucleotide identity, genetic relatedness, virulence determinants, and possible sites of reassortment. The results show that the 2008 H5N1 viruses displayed genetic drift characteristics (less than 3% genetic differences), as commonly found in influenza A viruses. Based on phylogenetic analysis, clade 1 viruses in Thailand were divided into 3 distinct branches (subclades 1, 1.1 and 1.2). Six out of 8 H5N1 isolates have been identified as reassorted H5N1 viruses, while other isolates belong to an original H5N1 clade. These viruses have undergone inter-lineage reassortment between subclades 1.1 and 1.2 and thus represent new reassorted 2008 H5N1 viruses. The reassorted viruses have acquired gene segments from H5N1, subclade 1.1 (PA, HA, NP and M) and subclade 1.2 (PB2, PB1, NA and NS) in Thailand. Bootscan analysis of concatenated whole genome sequences of the 2008 H5N1 viruses supported the reassortment sites between subclade 1.1 and 1.2 viruses. Based on estimating of the time of the most recent common ancestors of the 2008 H5N1 viruses, the potential point of genetic reassortment of the viruses could be traced back to 2006. Genetic analysis of the 2008 H5N1 viruses has shown that most virulence determinants in all 8 genes of the viruses have remained unchanged. In summary, two predominant H5N1 lineages were circulating in 2008. The original CUK2-like lineage mainly circulated in central Thailand and the reassorted lineage (subclades 1.1 and 1.2) predominantly circulated in lower-north Thailand. To prevent new reassortment, emphasis should be put on prevention of H5N1 viruses circulating in high risk areas. In addition, surveillance and whole genome sequencing of H5N1 viruses should be routinely performed for monitoring the genetic drift of the virus and new reassorted strains, especially in light of potential reassortment between avian and mammalian H5N1 viruses.

Reassortment

Subclade

Antigenic shift

Lineage (genetic)

H5N1 genetic structure

10.1186/1743-422x-7-233

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Citations (18)