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Complete genome sequences of H5N1 viruses derived from a domestic cat “A/Cat/Thailand/KU-02/04” and dog “A/Dog/Thailand/KU-08/04” were comprehensively analyzed and compared with H5N1 isolates obtained during the 2004 and 2005 outbreaks. Phylogenetic analysis of both cat and dog viruses revealed that they are closely related to the H5N1 viruses recovered from avian influenza outbreaks of the same period. Genetic analysis of 8 viral gene segments showed some evidence of virulence in mammalian species. In summary, the H5N1 viruses that infected a domestic cat and dog are highly pathogenic avian influenza viruses that are virulent in mammalian species, potentially indicating transmission of H5N1 viruses from domestic animals to humans. [ABSTRACT FROM AUTHOR]

Duck tembusu virus (DTMUV) is an emerging duck pathogen in China and other Asian countries. It is unclear whether this emerging zoonotic infection poses a threat to humans. A previous study in 2012 showed surprisingly high rates of seropositivity and positive viral detection by RT-PCR in duck farm workers in China. To understand the nature of the threat of this emerging virus, we studied the neutralizing antibody response to a local isolate of DTMUV in an at-risk population, who were workers in duck farms and residents around farming areas in Central Thailand where DTMUV had been previously detected, and in a not-at-risk population, who were people living in the same or neighbouring province, but at a distance from the farms and who had no contact with ducks. The sera from the at-risk population showed higher anti-DTMUV neutralizing antibody titres as compared with those of the not-at-risk population. However, within the at-risk population, workers with direct contact with ducks did not show higher neutralizing titres than those without direct contact. Interestingly, some people in the not-at-risk group also displayed high neutralizing antibody titres to DTMUV. These sera were tested against other endemic Flaviviruses and showed no or low cross-reactivity suggesting the specificity of the neutralizing activity against DTMUV. These data raise a possibility of DTMUV as a potential zoonotic pathogen but the mode of transmission of the virus from ducks or other possible hosts to humans should be explored further., (© 2021 Wiley-VCH GmbH.)

Zinc, Pb, Cd, Mn, Fe, Cr, and Cu levels in duck meat from large-scale farms have been found to be significantly higher than those from free-grazing duck farms. Zinc, Co, Mn, Cr, and Cu contamination levels in duck liver from large-scale farms were significantly higher than those from free-grazing farms; only Cd in duck liver from free-grazing farms was higher than in liver samples from large-scale farms at P < 0.05. Lead, Cd, Fe, and Cr levels in duck intestine samples from free-grazing farms were higher than large-scale farms at P < 0.001. Moreover, the average concentrations of Pb in duck meat and liver samples from large-scale farms and Cd levels in duck liver samples from free-grazing farm also exceeded the FAO/WHO and Codex Alimentarius limits by 100% (55/55), 100% (54/54), and 67.6% (23/34), respectively. PCA analysis showed a strong positive relationship between the eight metals in meat, liver, and intestine was > 0.69, > 0.69, and > 0.72, in order. The relationship of the liver combined with the intestine was > 0.65. This study indicated that consumers may incur health risks from long-term consumption of duck due to high Pb and Cd concentrations from both types of farms, particularly from large-scale duck farms.

The aim of this study was to determine the levels of Pb, Cd, Co and Cr in duck eggs and duck meat and to assess the risk of carcinogenic and non-carcinogenic effects caused by the consumption of duck products collected in Thailand. The human health risk assessment refers to the formulation of the USEPA standard focused on Estimated Daily Intake (EDI), Incremental Lifetime Cancer Risk (ILCR), Target Hazard Quotient (THQ) and Total Target Hazard Quotient (TTHQ). Ninety-eight percent of duck egg samples in this study were contaminated with Pb, with the average level typically above the standard limit. The EDI evaluation of heavy metal in children was the highest for all metals in the eggs. The calculated ILCR levels of Pb, Cd and Cr in eggs and meat consumption were higher than 10 -4 in children, adults, males, and females. Children were at risk when consuming contaminated duck eggs and the risk was higher than in adults by 3.9 times for Pb, Cd and Cr. This finding suggests that there is a high probability of cancer risk, particularly for children, which is higher than adults by absorption of these carcinogenic heavy metals through eggs. The calculation of THQ if consuming heavy metals contaminated duck eggs and meat in human exposure (70years) could conclude that the THQ male was higher than the female for all metals. However, the calculations of TTHQ for Pb, Cd, Co and Cr contamination in a duck egg and meat scenario for adults, males and females were still below the standard limit of 1, which means there was no risk, although there could be adverse health effects. This study suggested that children should be the focus of long-term monitoring for evaluating the carcinogenicity of these metals, more specifically in males., (Copyright © 2018. Published by Elsevier B.V.)

The cross-sectional study of heavy metals and aflatoxins was conducted by studying duck egg, blood, liver, feed, and water samples from 11 free grazing farms. The duck feed, soil, and rice samples were lower than the standard limit for aflatoxins at 20 ng g -1 . The highest levels of aflatoxin were detected in the liver, intestine, and egg yolk; these were 3.04, 1.73, and 4.56 ng g -1 respectively, while the average levels were 1.6 ± 0.73, 1.31 ± 0.23, and 1.87 ± 0.99 ng g -1 , respectively. Interestingly, aflatoxin was not detected in any of the egg white albumin samples (90/90 samples), whereas aflatoxin contamination was predominantly found in the yolk. Additionally, the means and standard deviations of Pb and Cd found in the duck eggs were 118.61 ± 22.25 and 13.84 ± 9.06 ng g -1 respectively. The maximum levels of Pb and Cd in eggs were 134.34 and 29.72 ng g -1 , respectively. Moreover, Pb contamination occurred in the duck liver tissue in 93.61% (88/94) of the samples, whereas Cd contamination was found in 100% (100/100). The maximum levels of Pb and Cd in the duck liver tissues were 606.16 and 22,946.20 ng g -1 , which was higher than those in the eggs. Pb contamination in the water from rice paddy fields after grazing was significantly higher than that in water samples taken before grazing. This is the first report of Pb and Cd contamination in blood, egg, and liver samples of free grazing ducks in Thailand.

This was a comparative study of the heavy metal levels (Zn, Pb, Cd, Co, Mn, Fe, Cr and Cu) in eggs from free grazing duck, small-scale, and large-scale farms in central and western regions of Thailand. A questionnaire was used to gather demographic data for the analysis of heavy metal contamination in feed, drinking water and wastewater. The correlation between the amounts of heavy metal contamination in eggs was studied against the heavy metals found in feed, drinking water and wastewater. The levels of Pb, Cd, Cr and Cu in eggs from large-scale farms were significantly higher than small farms and free grazing farms at P < 0.001. Zn in eggs from free grazing farms was higher than in the small farms and large-scale farms sampled. The contamination of Pb in eggs from all types of farms exceeded the standard limits of ACFS 6703-2005. The average levels of Pb in the eggs from small-scale farms correlated significantly with the level of Pb found in the feed at P < 0.05, while the average levels of Pb in eggs from free grazing duck farms correlated significantly with the levels of Pb found in the drinking water at P < 0.05. Additionally, the average level of Cu in duck egg from large-scale farms correlated significantly with the level of Cu found in the feeds at P < 0.001. Furthermore, from a calculation of the provisional tolerable daily intake (WHO-FAO) of heavy metals in this study, it was concluded that consumers face health risks from Cd contamination. Thus, heavy metal contamination, especially Pb and Cd in duck egg, must be of concern due to the health risks and the route of crucial heavy metals contamination should be elucidated and long - term monitoring of heavy metals posing health effects in farm systems should be carried out., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Highly pathogenic avian influenza (HPAI) H5N1 viruses have seriously affected the Asian poultry industry since their occurrence in 2004. Thailand has been one of those countries exposed to HPAI H5N1 outbreaks. This project was designed to compare the molecular evolution of HPAI H5N1 in Thailand between 2004 and 2008. Viruses with clade 1 hemagglutinin (HA) were first observed in early 2004 and persisted until 2008. Viruses with clade 2.3.4 HA were first observed in the northeastern region of Thailand between 2006 and 2007. Phylogenetic analysis among Thai isolates indicated that clade 1 viruses in Thailand consist of three distinct lineages: CUK2-like, PC168-like, and PC170-like viruses. The CUK2-like virus represents the predominant lineage and has been circulating throughout the course of the 4-year outbreaks. Analysis of recently isolated viruses has shown that the genetic distance was slightly different from viruses of the early outbreak and that CUK2-like viruses comprise the native strain. Between 2005 and 2007, PC168-like and PC170-like viruses were first observed in several areas around central and lower northern Thailand. In 2008, viruses reassorted from these two lineages, PC168-like and PC170-like viruses, were initially isolated in the lower northern provinces of Thailand and subsequently spread to the upper central part of Thailand. On the other hand, CUK2-like viruses were still detected around the lower northern and the upper central part of Thailand. Furthermore, upon emergence of the reassorted viruses, the PC168-like and PC170-like lineages could not be detected, suggesting that the only predominant strains still circulating in Thailand were CUK2-like and reassorted viruses. The substitution rate among clade 1 viruses in Thailand was lower. The virus being limited to the same area might explain the lower nucleotide substitution rate. This study has demonstrated that nationwide attempts to monitor the virus may help curb access and propagation of new HPAI viral genes.

This study was conducted to investigate space and time clusters of highly pathogenic avian influenza A (H5N1) virus infection and to determine risk factors at the subdistrict level in Thailand. Highly pathogenic avian influenza A (H5N1) was diagnosed in 1890 poultry flocks located in 953 subdistricts during 2004-2007. The ecologic risk for H5N1 virus infection was assessed on the basis of a spatial-based case-control study involving 824 case subdistricts and 3296 control subdistricts from 6 study periods. Risk factors investigated in clustered areas of H5N1 included human and animal demographic characteristics, poultry production systems, and wild birds and their habitats. Six variables remained statistically significant in the final model: flock density of backyard chickens (odds ratio [OR], 0.98), flock density of fighting cocks (OR, 1.02), low and high human density (OR, 0.60), presence of quail flocks (OR, 1.21), free-grazing duck flocks (OR, 2.17), and a poultry slaughterhouse (OR, 1.33). We observed a strong association between subdistricts with H5N1 virus-infected poultry flocks and evidence of prior and concomitant H5N1 infection in wild birds in the same subdistrict.

The aim of this study was to determine the performance of real-time amplification based methods - NASBA, TaqMan, RT-FRET, and RT-PCR LUXtrade mark formats - for the detection of influenza A (H5N1) virus RNA. In an analysis of 54 samples obtained from a range of animal species in Thailand during the period 2003-2006, results showed that the NASBA (H5=98.2%, N1=96.3%), TaqMan (H5=98.2%, N1=96.3%) and FRET (H5=98.2%, N1=96.3%) had significantly higher rates of positive detection than LUX (H5=94.4%, N1=50.0%; P<0.001) for influenza A, H5 and N1 isolates. There were no false-positive results from any methods used in the negative-control group of samples. The limits of analytical detection were at least 10copies/reaction in real-time NASBA and LUX assays, while FRET and TaqMan assay appeared to be less sensitive at > or =100copies/reaction. The assays were relatively specific without cross-reactivity to a number of other influenza strains or viral pathogens. In conclusion, our study demonstrated that real-time NASBA, TaqMan and FRET assays can be used to detect influenza A (H5N1) from a wide range of hosts, and be specific for H5N1 samples obtained during different outbreaks (2003-2006). All assays provided the benefit of rapid influenza H5N1 identification for early diagnosis, in the range of hours, and they are well suited to high throughput analyses.

This present study is the first to quantify the transmission of avian influenza virus H5N1 within flocks during the 2004 epidemic in Thailand. It uses the flock-level mortality data to estimate the transmission-rate parameter ( beta ) and the basic reproduction number (R(0)). The point estimates of beta varied from 2.26/day (95% confidence interval [CI], 2.01-2.55) for a 1-day infectious period to 0.66/day (95% CI, 0.50-0.87) for a 4-day infectious period, whereas the accompanying R(0) varied from 2.26 (95% CI, 2.01-2.55) to 2.64 (95% CI, 2.02-3.47). Although the point estimates of beta of backyard chickens and fighting cocks raised together were lower than those of laying hens and broiler chickens, this difference was not statistically significant. These results will enable us to assess the control measures in simulation studies. They also indicate that, for the elimination of the virus, a critical proportion of the susceptible poultry population in a flock (i.e., 80% of the population) needs to be vaccinated.

In 2005, total of 140 samples of duck meat and intestine from slaughterhouses in Nakhon Pathom Province, Thailand, were analyzed for Campylobacter spp. Twenty-eight samples (20%) were positive for Campylobacter spp using the standard culture method (SCM) with 21 samples of C. jejuni and the other 7 C. coli. Forty-four samples (31%) were positive using multiplex polymerase chain reaction, with 34 samples of C. jejuni and 10 of C. coli. This is the first report of Campylobacter contamination in duck in Thailand.

Outbreaks of H5N1 highly pathogenic avian influenza (HPAI) occurred in various types of domestic poultry in Thailand during 2004-05. H5N1 viruses were also detected in humans and other mammalian species. Infections were mainly detected in backyard chickens and domestic ducks. The geographic distribution of the 2004 outbreaks was widespread throughout Thailand; most outbreaks occurred in the Central Region, the southern part of the Northern Region, and the Eastern Region. In 2005, the H5N1 outbreaks continued and showed a clustered pattern in four provinces in the southern part of the Northern Region and in one province in the Central Region. H5N1 HPAI outbreaks caused serious socioeconomic consequences to the poultry industry, the social community, farmers' livelihood, and human health. After key measures were implemented, the incidence of the outbreaks declined remarkably in 2005.

Three major outbreaks of avian influenza (AI) occurred in Thailand. During the third episode in October 2005, we have isolated H5N1 viruses from one human case and three poultry cases. The whole genomes of AI viruses from human, chickens and quail from the outbreaks were characterized. Sequence analysis of eight gene segments revealed that the 2005 H5N1 viruses isolated in October 2005 were closely related to those recovered from chicken, tiger(s) and human(s) in January and July 2004. In addition, the genetic changes of the AI isolates at the HA cleavage site have been observed.

We report H5N1 virus infection in a domestic cat infected by eating a pigeon carcass. The virus isolated from the pigeon and the cat showed the same cluster as the viruses obtained during the outbreak in Thailand. Since cats are common house pets, concern regarding disease transmission to humans exists.

In addition to causing 12 human deaths and 17 cases of human infection, the 2004 outbreak of H5N1 influenza virus in Thailand resulted in the death or slaughter of 60 million domestic fowl and the disruption of poultry production and trade. After domestic ducks were recognized as silent carriers of H5N1 influenza virus, government teams went into every village to cull flocks in which virus was detected; these team efforts markedly reduced H5N1 infection. Here we examine the pathobiology and epidemiology of H5N1 influenza virus in the 4 systems of duck raising used in Thailand in 2004. No influenza viruses were detected in ducks raised in "closed" houses with high biosecurity. However, H5N1 influenza virus was prevalent among ducks raised in "open" houses, free-ranging (grazing) ducks, and backyard ducks.

The H5N1 avian influenza virus outbreak among zoo tigers in mid-October 2004, with 45 animals dead, indicated that the avian influenza virus could cause lethal infection in a large mammalian species apart from humans. In this outbreak investigation, six H5N1 isolates were identified and two isolates (A/Tiger/Thailand/CU-T3/04 and A/Tiger/Thailand/CU-T7/04) were selected for whole genome analysis. Phylogenetic analysis of the 8 gene segments showed that the viruses clustered within the lineage of H5N1 avian isolates from Thailand and Vietnam. The hemagglutinin (HA) gene of the viruses displayed polybasic amino acids at the cleavage site, identical to those of the 2004 H5N1 isolates, which by definition are highly pathogenic avian influenza (HPAI). In addition, sequence analyses revealed that the viruses isolated from tigers harbored few genetic changes compared with the viruses having infected chicken, humans, tigers and a leopard isolated from the early 2004 H5N1 outbreaks. Sequence analyses also showed that the tiger H5N1 isolated in October 2004 was more closely related to the chicken H5N1 isolated in July than that from January. Interestingly, all the 6 tiger H5N1 isolates contained a lysine substitution at position 627 of the PB2 protein similar to the human, but distinct from the original avian isolates.

In January 2004, highly pathogenic avian influenza (HPAI) virus of the H5N1 subtype was first confirmed in poultry and humans in Thailand. Control measures, e.g., culling poultry flocks, restricting poultry movement, and improving hygiene, were implemented. Poultry populations in 1,417 villages in 60 of 76 provinces were affected in 2004. A total of 83% of infected flocks confirmed by laboratories were backyard chickens (56%) or ducks (27%). Outbreaks were concentrated in the Central, the southern part of the Northern, and Eastern Regions of Thailand, which are wetlands, water reservoirs, and dense poultry areas. More than 62 million birds were either killed by HPAI viruses or culled. H5N1 virus from poultry caused 17 human cases and 12 deaths in Thailand; a number of domestic cats, captive tigers, and leopards also died of the H5N1 virus. In 2005, the epidemic is ongoing in Thailand.