Seroprevalence of Orientia tsutsugamushi and Rickettsia typhi in water buffaloes (Bubalus bubalis) from Southern Thailand.

Background and Aim: Scrub typhus and murine typhus are globally distributed zoonoses caused by the intracellular Gram-negative bacteria Orientia tsutsugamushi and Rickettsia typhi, respectively. Numerous studies have been undertaken on rickettsial illnesses in humans and animals, including arthropod vectors, in Thailand. However, the reports on the seroprevalence of antibodies to O. tsutsugamushi and R. typhi in buffaloes is extremely rare. Thus, this study aimed to estimate the seroprevalence of both rickettsial infections in water buffaloes (Bubalus bubalis) in Phatthalung Province, southern Thailand. Materials and Methods: From February to March 2023, a total of 156 serum samples were collected from 156 water buffaloes on 29 farms in Phatthalung province. The sera were screened for antibodies against O. tsutsugamushi and R. typhi using an indirect immunofluorescence assay. Results: The seroprevalence of antibodies against O. tsutsugamushi and R. typhi in individual water buffaloes was 4.49% (95% confidence interval [CI]: 2.19%-8.97%) and 3.85% (95% CI: 1.77%-8.14%), respectively, whereas 31% (9/29) of the herds had buffaloes with antibodies. The number of buffaloes with scrub typhus infection and ectoparasite infestation was statistically significant (p < 0.05; odds ratio = 6.25 [95% CI: 1.19-33.33]). Intriguingly, the prevalence of scrub typhus antibodies in buffaloes that were not infested with ectoparasites was much higher than those that were. Conclusion: This is the first report of O. tsutsugamushi and R. typhi antibodies in water buffalo sera in Southern Thailand. Two serum samples showed a high antibody titer against O. tsutsugamushi. Seroprevalence mainly occurred in non-ectoparasite-infested buffaloes, especially for O. tsutsugamushi antibodies. At the herd level, one-third of the studied farms showed seroprevalence. Additional research on the occurrence of these pathogens in vectors and in other animal reservoirs is necessary.

Bartonella vinsonii subsp. arupensis in humans, Thailand.

We identified Bartonella vinsonii subsp. arupensis in pre-enriched blood of 4 patients from Thailand. Nucleotide sequences for transfer-messenger RNA gene, citrate synthase gene, and the 16S-23S rRNA internal transcribed spacer were identical or closely related to those for the strain that has been considered pathogenic since initially isolated from a human in Wyoming, USA.

First report of three novel Bartonella species isolated in rodents and shrews from nine provinces of Thailand.

Background and Aim: Bartonella spp. are Gram-negative zoonotic bacteria that are transmitted to humans by several types of animal hosts, including rodents. Several studies have been conducted on the prevalence of Bartonella infections in rodents. However, the risk of rodent-associated Bartonella spp. infection in humans remains unclear. This study aimed to estimate the prevalence and genetic heterogeneity of Bartonella spp. in rodents and shrews from nine provinces of Thailand using culture and molecular techniques. Materials and Methods: A total of 860 blood samples from rodents and shrews across nine provinces of Thailand were collected from January 2013 to June 2016. Bartonella spp. were isolated from all samples using conventional culture techniques and polymerase chain reaction. Phylogenetic tree analysis was used to align the Bartonella sequences obtained from this study. Results: The prevalence of Bartonella spp. in rodents and shrews was 11.5% (99/860, 95% confidence interval: 9.38-13.64%). The following nine species of Bartonella were detected: Bartonella tribocorum, Bartonella rattimassiliensis, Bartonella queenslandensis, Bartonella elizabethae, Bartonella chanthaburi spp. nov., Bartonella satun spp. nov., Bartonella coopersplainsensis, Bartonella ranong spp. nov., and Bartonella henselae. The prevalence of Bartonella-positive animals differed significantly among provinces. Conclusion: To the best of our knowledge, the three novel Bartonella spp. isolated from rodents and shrews across Thailand were detected for the first time in this study. Further studies on the epidemiology of Bartonella infection in rodents and its interaction with human health should be conducted in accordance with the Thai government's "One Health" approach to humans, animals, and the environment.

The first report of seroprevalence of Q fever in water buffaloes (Bubalus bubalis) in Phatthalung, Thailand.

BACKGROUND AND AIM: Q fever is a worldwide zoonosis caused by the intracellular bacterium, Coxiella burnetii. A few studies focused on the occurrence of Q fever infection in water buffaloes in Thailand have been conducted; however, little is known regarding the seroprevalence of C. burnetii antibodies in buffaloes. In the present study, we describe the prevalence of Q fever infection in water buffaloes (Bubalus bubalis) in Phatthalung, Thailand. MATERIALS AND METHODS: A total of 421 samples (156 blood, 156 sera, and 109 ectoparasites [lice]) were collected from 156 water buffaloes from 29 farms of the Phatthalung Province from January 22, 2021, to March 26, 2021. The blood and ectoparasite samples were screened for C. burnetii DNA using a polymerase chain reaction assay and the sera were tested for C. burnetii antibody using an indirect immunofluorescence assay. RESULTS: C. burnetii DNA was not detected in blood or ectoparasites; however, the seroprevalence of individual water buffaloes was 4.49% (95% CI: 2.19-8.99%), whereas that of the herd was 13.79%. There was a significant difference between abortion history and Q fever infection at 29 farms (p=0.005; OR=33.55 [95%CI: 156-722.38]). CONCLUSION: This is the first report describing the low seroprevalence of C. burnetii antibodies in water buffaloes in Phatthalung Province, Thailand. The occurrence of this pathogen in buffaloes with reproductive disorders and people working with buffaloes warrant further investigation. Animal health authorities should inform farmers to effectively prevent and control this zoonosis.

The first report of the seroprevalence of antibodies against Bartonella spp. in water buffaloes (Bubalus bubalis) from South Thailand.

BACKGROUND AND AIM: Bartonellosis is an emerging worldwide zoonosis caused by bacteria belonging to the genus Bartonella. Several studies have been conducted on the prevalence of Bartonella infections from animals and humans, including reports from wild and domestic ruminants. However, there has been only one report of Bartonella infection in water buffaloes from the northeastern part of Thailand. Moreover, the seroprevalence of Bartonella spp. in water buffaloes still remains unknown. This study was conducted to explore the prevalence of Bartonella spp. among water buffaloes from South Thailand using molecular and serological techniques. MATERIALS AND METHODS: A total of 312 samples (156 blood and 156 sera) of 156 water buffaloes from 29 farms in Phatthalung Province, South Thailand, were collected from January to March 2021. All samples were screened for Bartonella spp. using polymerase chain reaction and indirect immunofluorescence assay. RESULTS: The seroprevalence of antibodies against three Bartonella spp. was 16.03% (25/156, 95% confidence interval: 10.65-22.74%), and among 25 water buffaloes with seroprevalence, 56%, 20%, and 24% were positive for antibodies against Bartonella henselae, Bartonella vinsonii subspp. berkhoffii, and Bartonella tamiae, respectively. No significant difference was detected among seroprevalence, gender, age, and ectoparasite infestation. CONCLUSION: This is the first report of the seroprevalence of antibodies against B. henselae, B. vinsonii subspp. berkhoffii, and B. tamiae in water buffaloes from South Thailand. Further studies are required on the epidemiology of Bartonella infection among water buffaloes, related personnel, and ectoparasites.

Salmonella prevalence in slaughtered buffaloes and pigs and antimicrobial susceptibility of isolates in Vientiane, Lao People's Democratic Republic.

This is the first report regarding isolation of Salmonella from cecum samples of buffaloes and pigs and characterization of the isolates in Laos. The organisms were isolated from 8% (4/50) of buffaloes and 76% (37/49) of pigs. In buffaloes, 3 animals harbored serotype 9,12: -:1,5, and 1 animal harbored both S. Derby and S. Javiana. In pigs, the most predominant serotypes were S. Derby (51%) followed by S. Anatum (45%), S. Weltevreden (15%) and S. Stanley (5%). The buffalo isolates were susceptible to the antimicrobials tested, whereas the pig isolates showed 10 resistance patterns to 1-5 antibiotics. Of the 59 pig isolates, the resistance rates to tetracycline, streptomycin, ampicillin, sulfamethoxazole-trimethoprim, chloramphenicol, amoxicillin-clavulanic acid and nalidixic acid were 24%, 22%, 14%, 5%, 2%, 2% and 2%, respectively. The results suggest that pigs and buffaloes harbor Salmonella, with a higher prevalence especially in pigs, and all the isolates showed sensitivity to cefotaxime, norfloxacin and ciprofloxacin.

The epidemiological relationship between Salmonella enterica serovar typhimurium and Salmonella enterica serovar 4,[5],12:i:- isolates from humans and swine in Thailand.

A total of 138 isolates of S. Typhimurium and S. 4,[5],12:i:- from humans and swine in Thailand during 2003-2006, were evaluated for antimicrobial sensitivity by the disk diffusion method against 10 antimicrobial drugs and pulsed-field gel electrophoresis (PFGE) with endonuclease Xbal to investigate the epidemiological relationship among isolates. It was found that all isolates were classified into 27 antimicrobial resistance patterns, and 80% of S. Typhimurium and 95.4% of S. 4,[5],12:i:- isolates were resistant to three or more antimicrobial agents. By PFGE testing, the 84 PFGE patterns were categorized into A to Z patterns. Eighty percent of S. Typhimurium and 71.3% of S. 4,[5],12:i:- isolates in 7 major PFGE patterns had close clonal relationships (_85% similarity). Our studies indicate the spread of genetically identical clones of S. Typhimurium and S. 4,[5],12:i:- in humans and swine in Thailand.

Isolation and phylogenetic analysis of Bartonella species from Rusa deer (Rusa timorensis) in Thailand.

The aim of the present study is to investigate the prevalence of Bartonella infection in deer in Thailand and to characterize the isolates by biochemical, morphological and genetic analysis. A total of 247 blood samples were collected from Rusa deer (Rusa timorensis) in a livestock breeding facility in Thailand. Bartonella bacteria were isolated in 3.6% of the blood samples. Three out of 110 (2.7%) males and 6 of 137 (4.4%) females were positive for Bartonella. A higher prevalence of Bartonella was observed in young deer under 4 years of age compared to adults over 4 years of age, but no Bartonella was isolated from deer over 8 years of age. Phylogenetic analysis of concatenated sequences of seven loci of Bartonella indicated that all the isolates from Rusa deer in Thailand were identical and formed a distinct cluster from other known Bartonella species.

Prevalence of Campylobacter spp. in slaughtered cattle and buffaloes in Vientiane, Lao People's Democratic Republic.

This is the first report regarding isolation of Campylobacter in caecum and bile samples obtained from ruminants in Vientiane, Lao PDR. Campylobacter was isolated from 3 (1.6%) of the 184 caecum samples and 1 (1.0%) of the 100 bile samples obtained from buffaloes. Three of the 4 isolates were determined to be C. jejuni, which was detected in 2 caecum samples and 1 bile sample; the other caecum sample contained C. fetus. Campylobacter was not isolated from any of the 82 cattle caecum samples. Our results suggest that cattle and buffaloes may not be important sources of Campylobacter food poisoning in Lao PDR.

Detection of Campylobacter in duck using standard culture method and multiplex polymerase chain reaction.

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.

Diversity and phylogenetic relationships among Bartonella strains from Thai bats.

Bartonellae are phylogenetically diverse, intracellular bacteria commonly found in mammals. Previous studies have demonstrated that bats have a high prevalence and diversity of Bartonella infections globally. Isolates (n = 42) were obtained from five bat species in four provinces of Thailand and analyzed using sequences of the citrate synthase gene (gltA). Sequences clustered into seven distinct genogroups; four of these genogroups displayed similarity with Bartonella spp. sequences from other bats in Southeast Asia, Africa, and Eastern Europe. Thirty of the isolates representing these seven genogroups were further characterized by sequencing four additional loci (ftsZ, nuoG, rpoB, and ITS) to clarify their evolutionary relationships with other Bartonella species and to assess patterns of diversity among strains. Among the seven genogroups, there were differences in the number of sequence variants, ranging from 1-5, and the amount of nucleotide divergence, ranging from 0.035-3.9%. Overall, these seven genogroups meet the criteria for distinction as novel Bartonella species, with sequence divergence among genogroups ranging from 6.4-15.8%. Evidence of intra- and intercontinental phylogenetic relationships and instances of homologous recombination among Bartonella genogroups in related bat species were found in Thai bats.

Antimicrobial resistance in Campylobacter isolated from food animals and humans in northern Thailand.

A study was conducted to determine the prevalence of Campylobacter with antimicrobial resistance from chickens, pigs, dairy cows, healthy farm workers, and children hospitalized with diarrhea in northern Thailand. Resistance was highest in pig samples and lowest in healthy farm workers. Resistance to fluoroquinolones and tetracycline was high in all study populations. The increased prevalence of resistant isolates from meat samples collected at markets, compared to isolates collected from animals on the farm or the slaughterhouse, suggests that contamination after carcasses leave the slaughterhouse is an important factor in the spread of resistant bacteria into the human food chain.

Recovery of Salmonella using a combination of selective enrichment media and antimicrobial resistance of isolates in meat in Thailand.

From November 2004 to March 2005, 50 samples (chicken, pork and beef) of registered meat and non-registered meat were purchased from supermarkets and retail markets located in Bangkok, Thailand. Each sample was evaluated for Salmonella spp by a conventional method using combination of selective enrichment media (RV+MSRV) and compared with selective enrichment medium alone (DIASALM). Our study revealed the performance of RV+MSRV for the detection of Salmonella spp was significantly better than those of DIASALM alone since the recovery of Salmonella spp in both groups of meat was high using RV+MSRV, particularly in the registered meat. In addition, the recovery of serovars in registered meat was significantly higher than those in non-registered meat. Antimicrobial resistance of 62 Salmonella isolates in both groups of meat was determined for 10 antimicrobial drugs using the disk diffusion method. The results show that 100% of isolates from both groups were susceptible to amoxicillin/clavulanic acid, ciprofloxacin, cefotaxime and norfloxacin and 50-60% of isolates from both groups were resistant to tetracycline, streptomycin and ampicillin. Sixty percent of Salmonella isolates from meat showed multiresistance antimicrobial patterns.

Serotypes and antimicrobial resistance of Campylobacter jejuni isolated from humans and animals in Thailand.

We investigated the serotypes, distributions, and antimicrobial resistance of Campylobacter jejuni isolates from humans and animals as a source of infection in poultry between 2002 and 2003. A total of 50 C. jejuni isolates from humans and 29 C. jejuni isolates from poultry were studied for serotype using the Penner serotyping scheme and the drug susceptibilities of the isolates which were determined for 7 antimicrobial drugs using the disk diffusion method. Serotype B (10%), serotype E (8%) and serotype R (8%) were found in humans isolates, while serotype A (27%) was most freguently isolated from poultry, followed by serotype K (21%) and serotype C (13%). Resistance in human isolates to cephalothin was high (100%). Resistance to trimethoprim/sulfamethoxazole, ciprofloxacin, and nalidixic acid were observed in 90, 82 and 78% of isolates, respectively. Most of the isolates (88%) were susceptible to erythromycin. High levels of resistance to drugs (ciprofloxacin and nalidixic acid) were observed in the isolates from poultry. These results indicate the importance of poultry as a reservoir of C. jejuni infection in Thailand is limited. In addition, a high proportion of the isolates were resistant to antimicrobial drugs, particularly the quinolone group.

Prevalence of zoonotic Bartonella species among rodents and shrews in Thailand.

We investigated the prevalence of Bartonella species in 10 rodent and one shrew species in Thailand. From February 2008 to May 2010, a total of 375 small animals were captured in 9 provinces in Thailand. Bartonella strains were isolated from 57 rodents (54 from Rattus species and 3 from Bandicota indica) and one shrew (Suncus murinus) in 7 of the 9 provinces, and identified to the species level. Sequence analysis of the citrate synthase and RNA polymerase ß subunit genes identified the 58 isolates from each Bartonella-positive animal as B. tribocorum in 27 (46.6%) animals, B. rattimassiliensis in 17 (29.3%) animals, B. elizabethae in 10 (17.2%) animals and B. queenslandensis in 4 (6.9%) animals. R. norvegicus, R. rattus, and Suncus murinus carried B. elizabethae, which causes endocarditis in humans. The prevalence of Bartonella bacteremic animals by province was 42.9% of the animals collected in Phang Nga, 26.8% in Chiang Rai, 20.4% in Sa Kaeo, 16.7% in Nakhon Si Thammarat, 12.0% in Surat Thani, 9.1% in Mae Hong Son and Loei Provinces. These results indicate that Bartonella organisms are widely distributed in small mammals in Thailand and some animal species may serve as important reservoirs of zoonotic Bartonella species in the country.

The global one health paradigm: challenges and opportunities for tackling infectious diseases at the human, animal, and environment interface in low-resource settings.

Zoonotic infectious diseases have been an important concern to humankind for more than 10,000 years. Today, approximately 75% of newly emerging infectious diseases (EIDs) are zoonoses that result from various anthropogenic, genetic, ecologic, socioeconomic, and climatic factors. These interrelated driving forces make it difficult to predict and to prevent zoonotic EIDs. Although significant improvements in environmental and medical surveillance, clinical diagnostic methods, and medical practices have been achieved in the recent years, zoonotic EIDs remain a major global concern, and such threats are expanding, especially in less developed regions. The current Ebola epidemic in West Africa is an extreme stark reminder of the role animal reservoirs play in public health and reinforces the urgent need for globally operationalizing a One Health approach. The complex nature of zoonotic diseases and the limited resources in developing countries are a reminder that the need for implementation of Global One Health in low-resource settings is crucial. The Veterinary Public Health and Biotechnology (VPH-Biotec) Global Consortium launched the International Congress on Pathogens at the Human-Animal Interface (ICOPHAI) in order to address important challenges and needs for capacity building. The inaugural ICOPHAI (Addis Ababa, Ethiopia, 2011) and the second congress (Porto de Galinhas, Brazil, 2013) were unique opportunities to share and discuss issues related to zoonotic infectious diseases worldwide. In addition to strong scientific reports in eight thematic areas that necessitate One Health implementation, the congress identified four key capacity-building needs: (1) development of adequate science-based risk management policies, (2) skilled-personnel capacity building, (3) accredited veterinary and public health diagnostic laboratories with a shared database, and (4) improved use of existing natural resources and implementation. The aim of this review is to highlight advances in key zoonotic disease areas and the One Health capacity needs.

Global distribution of Bartonella infections in domestic bovine and characterization of Bartonella bovis strains using multi-locus sequence typing.

Bartonella bovis is commonly detected in cattle. One B. bovis strain was recently isolated from a cow with endocarditis in the USA, suggesting its role as an animal pathogen. In the present study, we investigated bartonella infections in 893 cattle from five countries (Kenya, Thailand, Japan, Georgia, and Guatemala) and 103 water buffaloes from Thailand to compare the prevalence of the infection among different regions and different bovid hosts. We developed a multi-locus sequence typing (MLST) scheme based on nine loci (16S rRNA, gltA, ftsZ, groEL, nuoG, ribC, rpoB, ssrA, and ITS) to compare genetic divergence of B. bovis strains, including 26 representatives from the present study and two previously described reference strains (one from French cows and another from a cow with endocarditis in the USA). Bartonella bacteria were cultured in 6.8% (7/103) of water buffaloes from Thailand; all were B. bovis. The prevalence of bartonella infections in cattle varied tremendously across the investigated regions. In Japan, Kenya, and the Mestia district of Georgia, cattle were free from the infection; in Thailand, Guatemala, and the Dusheti and Marneuli districts of Georgia, cattle were infected with prevalences of 10-90%. The Bartonella isolates from cattle belonged to three species: B. bovis (n=165), B. chomelii (n=9), and B. schoenbuchensis (n=1), with the latter two species found in Georgia only. MLST analysis suggested genetic variations among the 28 analyzed B. bovis strains, which fall into 3 lineages (I, II, and III). Lineages I and II were found in cattle while lineage III was restricted to water buffaloes. The majority of strains (17/28), together with the strain causing endocarditis in a cow in the USA, belonged to lineage I. Further investigations are needed to determine whether B. bovis causes disease in bovids.

Salmonella prevalence in meat at retail markets in Pakse, Champasak Province, Laos, and antimicrobial susceptibility of isolates.

Non-typhoidal Salmonella is one of the most common causes of human gastroenteritis worldwide and most human outbreaks are associated with the consumption of contaminated food. However, there are no reports on Salmonella contamination in market meat in Laos. The objective of this study was to determine the prevalence of Salmonella in meat samples in Pakse, Champasak Province, Laos, as well as the antimicrobial susceptibility of isolates. The prevalence of Salmonella was 82% in beef, 93% in pork and 80% in buffalo meat. In total, 80 isolates and 11 serovars were found, including serovars Stanley (n=15), Anatum (n=14), Derby (n=11), Rissen (n=9) and Amsterdam (n=7). The drug susceptibility of 60 strains against 10 antimicrobial agents was tested. The 60 isolates examined were sensitive to ciprofloxacin (100% susceptible), norfloxacin (100%), cefotaxime (95%), nalidixic acid (90%) and chloramphenicol (88%), but were resistant to streptomycin (67% resistant), tetracycline (67%) and ampicillin (63%). Of the isolates, 73% were multidrug-resistant. These findings indicate a high Salmonella prevalence in market meat in Pakse. Therefore, programmes to control Salmonella contamination are needed.

Enrichment culture and molecular identification of diverse Bartonella species in stray dogs.

Using pre-enrichment culture in Bartonella alpha-Proteobacteria growth medium (BAPGM) followed by PCR amplification and DNA sequence identification that targeted a fragment of the citrate synthase gene (gltA), we provide evidence of common bartonella infections and diverse Bartonella species in the blood of stray dogs from Bangkok and Khon Kaen, Thailand. The overall prevalence of all Bartonella species was 31.3% (60/192), with 27.9% (31/111) and 35.8% (29/81) in the stray dogs from Bangkok and Khon Kaen, respectively. Phylogenetic analyzes of gltA identified eight species/genotypes of Bartonella in the blood of stray dogs, including B. vinsonii subsp. arupensis, B. elizabethae, B. grahamii, B. quintana, B. taylorii, and three novel genotypes (BK1, KK1 and KK2) possibly representing unique species with ≤ 90.2% similarities to any of the known Bartonella species B. vinsonii subsp. arupensis was the only species detected in dogs from both sites, B. quintana and BK1 were found in the dogs from Bangkok, B. elizabethae, B. taylorii, KK1 and KK2 were found in the dogs from Khon Kaen. We conclude that stray dogs in Thailand are frequently infected with Bartonella species that vary with geographic region. As some Bartonella species detected in the present study are considered pathogenic for humans, stray dogs in Thailand may serve as possible reservoirs for Bartonella causing human illnesses. Further work is needed to determine the role of those newly discovered Bartonella genotypes/species in human and veterinary medicine.

Identification of Bartonella infections in febrile human patients from Thailand and their potential animal reservoirs.

To determine the role of Bartonella species as causes of acute febrile illness in humans from Thailand, we used a novel strategy of co-cultivation of blood with eukaryotic cells and subsequent phylogenetic analysis of Bartonella-specific DNA products. Bartonella species were identified in 14 blood clots from febrile patients. Sequence analysis showed that more than one-half of the genotypes identified in human patients were similar or identical to homologous sequences identified in rodents from Asia and were closely related to B. elizabethae, B. rattimassiliensis, and B. tribocorum. The remaining genotypes belonged to B. henselae, B. vinsonii, and B. tamiae. Among the positive febrile patients, animal exposure was common: 36% reported owning either dogs or cats and 71% reported rat exposure during the 2 weeks before illness onset. The findings suggest that rodents are likely reservoirs for a substantial portion of cases of human Bartonella infections in Thailand.