Intranasal immunization with a recombinant outer membrane protein H based Haemorrhagic septicemia vaccine in dairy calves.

Haemorrhagic septicemia (HS) is a contagious disease in cattle with high morbidity and mortality rates. HS vaccine in Thailand is an oil-adjuvant formulation, and is difficult to administer. The present study aimed to formulate and evaluate the protection in dairy calves conferred by immunization with an in-house intranasal HS vaccine. The intranasal vaccine was formulated in a total volume of 500 µl containing either 50 or 100 µg of the recombinant outer membrane protein H (rOmpH) of Pasteurella multocida strain M-1404 (serovar B:2), and 10 µg of Cytosine-phosphate-guanosine oligodeoxynucleotides (CpG-ODN) as a mucosal adjuvant. Intranasal immunizations were conducted three times at three-week intervals. The antibodies post-immunization were detected by indirect ELISA and demonstrated efficient in vitro activity in suppressing a P. multocida strain from the complement-mediated killing assay. An intranasal vaccine induced both the serum IgG and secretory IgA levels that were significantly higher than the level conferred by the parenteral vaccine (P<0.05). Challenge exposure was conducted with a P. multocida strain M-1404 at day 72 of the experiments. The immunized calves had reduced clinical signs after challenge exposure that would normally result in disease proliferation. We conclude that intranasal vaccination of calves with rOmpH with CpG-ODN 2007 stimulated serum and secretory antibodies to rOmpH and whole cells of P. multocida strain M-1404 antigen. Moreover, it would result in protection in calves against artificial P. multocida infection.

Non-invasive endocrine monitoring indicates seasonal variations in gonadal hormone metabolites in dholes (Cuon alpinus).

To date, there is no information on reproductive endocrinology of dholes (Cuon alpinus). The objectives of the present study were as follows: (i) to characterize longitudinal profiles of gonadal steroids; and (ii) to examine the relationship between gonadal hormones and sexual behaviours in dholes. Three breeding pairs and two bachelor males were included in the study. Among these, four animals (2 males and 2 females; 4 years old) were imported from The Netherlands to Thailand 3 months before the study onset; the remaining individuals (3 males and 1 female; 5-7 years old) were native born. Faecal samples were collected 3-7 days/week for 12 months, extracted and assessed for gonadal hormone metabolites using a validated enzyme immunoassay. Observations of behaviour were conducted in 30 min sessions, 3-5 days/week. For the three breeding males, testosterone was elevated (P < 0.05) from October to January in the two imported males, whereas the concentration of steroid metabolites was high from April to June and from September to November in the native male. However, there was no clear seasonal pattern of reproductive hormone in the bachelor group. Oestrogen metabolite level of imported females was elevated for 9-12 days in January, followed by a rise in progestagen concentration. For native females, oestrogen metabolites were above the basal values in April and September, each of which was followed by a rise in progestagen concentration that remained elevated for 77 and 112 days, respectively. Sexual behaviours, including solicitation, mounting and copulations, were observed during the oestrogen peak in all females. Our findings indicate that reproductive seasonality of dholes may depend on the animals' origin and social group.

Thailand Antimicrobial Resistance Containment and Prevention Program.

The Thailand Antimicrobial Resistance (AMR) Containment and Prevention Program was founded to develop, co-ordinate and implement AMR Containment and Prevention (AMRCP) operational actions in Thailand following the 'One Health' approach. This article summarises the ten AMRCP operational actions initiated during the initial phase of the programme from 2012 to 2016: estimating the national AMR burden; establishing the dynamics of AMR chains to understand how AMR in Thailand develops and spreads; developing a national AMRCP infrastructure; developing laboratory and information technology systems for surveillance of AMR, antibiotic use and hospital-acquired infections; regulating the use and distribution of antibiotics in humans and food animals; generating local evidence for promoting responsible use of antibiotics and efficient practices for infection prevention and control; designing AMRCP campaigns; creating an AMRCP package; implementing the AMRCP package in selected pilot communities; and conducting research and development on diagnostics, therapy and prevention of antimicrobial-resistant bacterial infections. The programme's core campaign is to stop producing AMR by promoting responsible use of antibiotics, and to stop the acquisition and transmission of AMR by promoting good sanitation and hygiene as well as compliance with infection control and prevention practices.

Assessment of ovarian activity in captive goral (Naemorhedus griseus) using noninvasive fecal steroid monitoring.

To date, there is no information on gonadal steroidogenic activity of female goral (Naemorhedus griseus), a threatened species of Thailand. Captive goral populations have been established to produce animals for ex situ conservation and reintroduction, but as yet none are self-sustaining. The objectives of the present study were to (1) determine the influence of season on ovarian steriodogenic function; and (2) examine the relationship between gonadal hormone excretion and sexual behaviors throughout the year. Fecal samples were collected 5 to 7 days/wk for 15 months from 8 adult females housed at Omkoi Wildlife Breeding Center in Thailand and analyzed for ovarian steroid metabolites using validated enzyme immunoassays. Observations of sexual behaviors and mating were conducted each morning for 30 min/session. Based on fecal estrogen and progestagen metabolite concentrations, the overall estrous cycle length was about 21 days, with a 2- to 3-day follicular phase and an 18- to 20-day luteal phase. Sexual behaviors, most notably tail-up, increased for 2 to 3 days during the time estrogens were elevated during mating. Fecal progestagens were elevated during luteal phases and increased further during gestation, which lasted approximately 7 months. The lactation period was 5 months, and females were anestrus for 2 to 5 of those months, with the exception of one that cycled continuously throughout. Two females conceived around 2 months postpartum and so were pregnant during lactation. Birth records over the past 21 years indicated young are born throughout the year. This combined with the hormonal data suggests that female gorals are not strongly seasonal, at least in captivity, although there was considerable variation among females in estrogen and progestagen patterns. In conclusion, fecal steroid metabolite monitoring is an effective means of assessing ovarian function in this species and will be a useful tool for breeding management and planned development of assisted reproductive techniques such as artificial insemination and embryo transfer.

Seasonality of fecal androgen and glucocorticoid metabolite excretion in male goral (Naemorhedus griseus) in Thailand.

There is no information on the endocrinology of Chinese goral (Naemorhedus griseus), a high priority species for captive breeding and reintroduction in Thailand. This study characterized fecal androgen and glucocorticoid metabolites in male goral at Omkoi Wildlife Sanctuary to investigate seasonal relationships. Fecal samples were collected 3 days/week for 1 year from eight adult males. Mean androgen metabolite concentrations were greater (P<0.05) during the rainy season (289.82±9.18ng/g) and winter (224.09±11.97ng/g) compared to the summer (195.48±8.23ng/g), and were related to breeding activity. A similar pattern was observed for glucocorticoid concentrations (22.10±0.72ng/g compared to 21.98±0.98ng/g compared to 15.30±0.48ng/g), respectively, and this resulted in a positive correlation between the two hormones (P<0.05). There also were positive correlations between fecal androgen metabolite concentrations and temperature (P<0.05) and day length (P<0.05). In summary, this is the first study to assess endocrine function in male goral, and results showed seasonal variation in testicular and adrenal steroidogenic function, with greater activity in the rainy season and winter. Given that resources for captive male goral are consistent throughout the year, reproduction may be regulated primarily by photoperiod in this species.

Gender, season and management affect fecal glucocorticoid metabolite concentrations in captive goral (Naemorhedus griseus) in Thailand.

Chinese goral (Naemorhedus griseus) are a threatened species in Thailand and the focus of captive breeding for possible reintroduction. However, little is known of their biology or what factors in the captive environment affect welfare. Our objective was to determine the impact of gender, season, and management on goral adrenal activity. We hypothesized that differences in fecal glucocorticoid concentrations would be related to animal density. Fecal samples were collected 3 days/week for 1 year from 63 individuals (n = 32 males, 31 females) at two facilities that house the majority of goral in Thailand: Omkoi Wildlife Sanctuary (Omkoi), an off-exhibit breeding center that houses goral in individual pens (16 pens; n = 8 males, 8 females) and in small family groups (8 pens; n = 8 males, 8 females); and the Chiang Mai Night Safari (NS), a zoo that maintains 31 goral (n = 17 males, 14 females) in one large pen. Glucocorticoid metabolite concentrations were higher in male than female goral at Omkoi throughout the year, and there was a seasonal effect on adrenal activity (p<0.05). Goral at Omkoi and NS were used to test the effect of animal density on fecal glucocorticoid excretion of goral housed in similar-sized enclosures. Overall, the highest levels were found at NS (n = 31 adults/pen; 27 m2 per animal) compared to Omkoi (n = 2 adults/pen; 400 m2 per animal) (p<0.05). Overall findings support our hypothesis that animal density and aspects of the captive environment impact adrenal steroid activity in captive goral. In addition, gender and season also had significant effects on glucocorticoid metabolite production. Potential stressors pertaining to the welfare of this species were identified, which will guide future efforts to improve management and create self-sustaining and healthy populations of this threatened species.

An integrative approach to enhancing small-scale poultry slaughterhouses by addressing regulations and food safety in northern -Thailand.

BACKGROUND: In Asian countries, small-scale rural poultry meat production can face challenges due to food safety policies that limit economic growth and hinder improvement of sanitation and disease prevention. In this study, an integrative, participatory research approach was used to elucidate the sanitation and disease prevention practices in small-scale poultry slaughterhouses in rural northern Thailand. METHODS: Initial steps included the identification of key stakeholders associated with the meat production chain, development of a research framework, and design of a methodology based on stakeholder consultations. The framework and methodology combine issues in five major areas: (1) public health, (2) socioeconomics, (3) policy, (4) veterinary medicine, and (5) communities and the environment. Methods used include questionnaires, direct observation, focus groups, and in-depth interviews. In addition, a microbiological risk assessment approach was employed to detect Salmonella contamination in meat processing facilities. The microbial risk assessment was combined with stakeholder perceptions to provide an overview of the existing situation, as well as to identify opportunities for upgrading slaughterhouses in order to more effectively address matters of food safety, processing, and government licensing. RESULTS: The conceptual framework developed elucidated the complex factors limiting small-scale slaughterhouse improvement including a lack of appropriate enabling policies and an apparent absence of feasible interventions for improvement. Unhygienic slaughterhouse management was reflected in the incidence of Salmonella contamination in both the meat and the surrounding environment. CONCLUSION: There is potential for the use of an integrative approach to address critical problems at the interface of rural development and public health. The findings of this study could serve as a model for transdisciplinary studies and interventions related to other similar complex challenges.

PREVALENCE AND ANTIMICROBIAL RESISTANCE OF SALMONELLA ISOLATED FROM CARCASSES, PROCESSING FACILITIES AND THE ENVIRONMENT SURROUNDING SMALL SCALE POULTRY SLAUGHTERHOUSES IN THAILAND.

Salmonella is a major food-borne pathogen worldwide, including Thai- land, and poultry meat plays a role as a vehicle for the spread of the disease from animals to humans. The prevalence and characteristics of Salmonella isolated from 41 small scale poultry slaughterhouses in Chiang Mai, Thailand were determined during July 2011 through May 2012. Salmonella's prevalence in live poultry, car- casses, waste water, and soil around processing plants were 3.2%, 7.3%, 22.0% and 29.0%, respectively. Eighteen different serotypes were identified, the most common being Corvallis (15.2%), followed by Rissen (13.9%), Hadar (12.7%), Enteritidis (10.1%), [I. 4,5,12:i:-] (8.8%), Stanley (8.8%), and Weltevreden (8.8%). Antimicrobial susceptibility tests revealed that 68.4% of the Salmonella spp were resistant to at least one antimicrobial while 50.6% showed multiple drug resis- tance (MDR). Specifically, 44.3% of Salmonella were resistant to nalidixic acid, followed by streptomycin (41.8%), ampicillin (34.2%), tetracycline (34.2%), and sulfamethoxazole/trimethoprim (20.3%). Salmonella contamination was found in processing lines, carcasses, and in the environment around the processing sta- tions. These findings indicate that improving hygiene management in small scale poultry slaughterhouses as well as prudent use of antimicrobial drugs is urgently needed if Salmonella contamination is to be reduced.

Investigation of dogs as a reservoir of Penicillium marneffei in northern Thailand.

BACKGROUND: Penicillium marneffei is a dimorphic pathogenic fungus endemic in Southeast Asia that usually causes disseminated disease, mainly in immunocompromised individuals, especially those with HIV infection. Untreated cases are usually fatal. The only known natural reservoir exists in bamboo rats and there is no firm evidence that these animals are involved in direct transmission to humans. The risk of infection is not restricted to those living in endemic areas; HIV-infected individuals who travel to Southeast Asia have also become infected by P. marneffei. Hence, there must exist sources to which even tourists are exposed on a short-term basis. DESIGN AND METHODS: Penicillium is known to infect dogs and this animal is common in the streets and temple areas of Chiang Mai, where there is one of the highest incidences of P. marneffei infection in the world. Dogs have not been well studied as a possible reservoir. To investigate this possibility, we took nasal swabs from 83 outdoor dogs and performed culture and nested polymerase chain reaction (PCR) to detect P. marneffei. RESULTS: We found that approximately 13% of nasal swabs from dogs in Chiang Mai, Thailand were positive when tested by two different PCR methods, but culture results were negative. Sequencing the products from both PCR reactions showed 100% identity with P. marneffei, whereas no other known fungi shared both sequences. CONCLUSIONS: Our results suggest that dogs might be an animal reservoir for P. marneffei in northern Thailand. This observation should be confirmed by additional studies.

Induction of the ovulatory LH surge in Asian elephants (Elephas maximus): a novel aid in captive breeding management of an endangered species.

A unique feature of the reproductive physiology of Asian elephants (Elephas maximus) is the occurrence of two LH surges before ovulation, instead of one. An anovulatory LH (anLH) surge, the function of which is unknown, occurs consistently 3 weeks before the ovulatory LH (ovLH) surge that induces ovulation. Thus, the ability to induce an ovLH surge would be useful for scheduling natural mating or artificial insemination. The present study tested the efficacy of a gonadotrophin-releasing hormone agonist (GnRH-Ag) to induce LH surges during the follicular phase of the oestrous cycle, which resulted in varied LH responses, but generally none were as high as previously documented natural surges. Thus, for the ovulation-induction trials, nine females were administered 80 microg GnRH-Ag intravenously at three time periods during the oestrous cycle, namely the anovulatory follicular phase, the ovulatory follicular phase and the luteal phase. During the late anovulatory follicular phase, nine of 10 females (90%) responded with an immediate LH surge followed 15-22 days later by an ovLH surge or a post-ovulatory increase in progestagens. In contrast, despite responding to the GnRH-Ag with an immediate increase in LH, none of the females treated during other periods of the oestrous cycle exhibited subsequent ovLH surges. One cow got pregnant from natural mating following the induced ovLH surge. In conclusion, ovLH induction is possible using a GnRH-Ag, but only during a specific time of the anovulatory follicular phase.

Ecologic risk factor investigation of clusters of avian influenza A (H5N1) virus infection in Thailand.

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.

Use of genital inspection and female urine tests to detect oestrus in captive Asian elephants.

Captive Asian elephant (Elephas maximus) populations are decreasing due to low birth rates compared to wild elephants. Improving oestrous detection in female elephants is required to ensure successful mating in captive and semi-captive herds. Responsive behaviours of eight semi-captive bull elephants to the uro-genital area (genital inspection test) or urinary pheromones (urine test) of 14 female elephants throughout the oestrous cycle were evaluated. Weekly blood samples were collected for 27 consecutive months (14 months for the genital inspection test and 13 months for the urine test) from female elephants to characterize the patterns of circulating progestagen. Responsive behaviours of bulls were compared between females in the follicular versus the luteal phase of the cycle. The sensitivity and specificity of the genital inspection test were 65% and 68%, while those of the urine test were 52% and 61%, respectively. The bulls showed significantly higher "genital inspection", "flehmen from genital area" and "trunk on back" behaviours during the genital inspection test, and "flehmen" behaviours during the urine test in oestrous than in non-oestrous females. In sum, this study showed that monitoring sexual behaviours of Asian elephant bulls towards females or their urine can be used to detect the oestrous period. Although the sensitivity and specificity of both tests were not as high as expected, still, these methods appear to be more efficient at detecting oestrous than traditional methods based on mahout estimations of female receptivity. The use of genital inspection and urine tests may lead to more successful matings and thus to creating self-sustaining populations of captive elephants in range countries.

Identification of virulence-associated antigens and plasmids in Rhodococcus equi from patients with acquired immune deficiency syndrome and prevalence of virulent R. equi in soil collected from domestic animal farms in Chiang Mai, Thailand.

The prevalence of virulent Rhodococcus equi in soil collected from 17 domestic animal farms (from 12 cattle, 1 pig, and 4 horse farms) and in 6 clinical specimens from patients with acquired immune deficiency syndrome (AIDS) in Chiang Mai, Thailand, was investigated. The isolates were tested for the presence of 15-17-kDa antigens (VapA) and a 20-kDa antigen (VapB) by immunoblotting and for the presence of virulence plasmid DNA. Rhodococcus equi was isolated from most soil samples (68 of 80) obtained from the 17 farms, with 2.0 x 10(2) to 6.0 x 10(5) colony-forming units per gram of soil. We detected VapA in none of the 537 isolates from the soil samples. In one isolate from a pig farm, both VapB and virulence plasmid DNA were detected. Of the 6 clinical isolates from patients with AIDS, however, 4 isolates contained both VapB and virulence plasmid DNA. The remaining 2 isolates were avirulent.