The infection of Cysticercus fasciolaris in natural rats (Rattus species) residing in human residence areas, Nakhon Si Thammarat, Thailand.

Cysticercus fasciolaris (C. fasciolaris) is the larval stage of a cestode parasite named Taenia taeniaeformis (T. taeniaeformis). C. fasiolaris is found in small rodents, especially rats. Rattus species are listed as intermediate hosts of this parasite, and cats are the main definitive host of C. fasiolaris. The objective of this study was to study the pathological, microscopic, and molecular aspects of C. fasciolaris in rodents residing in human residence areas. One hundred and two rodents were trapped in human settlements and dissected for larva-containing cyst examinations in the body cavity. The larvae of C. fasciolaris were investigated using histopathological examination, microscopic observations under a stereomicroscope and scanning electron microscope, and molecular detection using polymerase chain reaction. The prevalence of hepatic cysts containing larvae was 8.91% (95% CI = 4.16-16.24). In addition, the older larvae also had longer micropapillae. Histopathological investigation revealed normal hepatic tissue containing larvae and a scanty fluid cyst. The cyst capsule contains mostly mononuclear cells and spindle cells in all infected rats. The molecular detection using two primer sets revealed the amplicons were similar to the clade of C. fasciolaris. In the future, more investigation is necessary to fully understand the parasite's molecular pathogenesis and virulent molecules, which are less obvious.

Antibacterial and antivirulence factor activities of protein hydrolysates from Phatthalung Sangyod rice (Oryza sativa L.) seeds against zoonotic and foodborne pathogens.

Background and Aim: Antimicrobial resistance is an emerging public health threat. Foodborne illnesses are typically caused by bacteria, such as Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus, and Staphylococcus aureus, which are frequently resistant to common antimicrobial agents. Rice is a staple grain in most parts of the world. Our previous work showed that Phatthalung Sangyod rice seed protein hydrolysates (SYPs), especially SYP4, exhibit antifungal activity against several fungal species that are pathogenic for both humans and animals and are non-cytotoxic to animal red blood cells. In this study, we aimed to determine the effects of the bioactive peptides in SYPs against several pathogenic bacteria in humans and animals. Materials and Methods: After isolating SYP1, it was treated as follows: heated (SYP2), and hydrolyzed using pepsin (SYP3), and proteinase K (SYP4). Then, we used 500 µg of protein to evaluate the antibacterial effects on four pathogenic bacteria, including E. coli, P. aeruginosa, B. cereus, and S. aureus, using agar well diffusion. Using a broth microdilution assay, we determined the minimum inhibitory and bactericidal concentration (MIC and MBC, respectively) values of active SYPs. Using the agar well diffusion and microtube incubation methods, we also assessed the inhibitory effects of SYPs on the bacterial quorum sensing (QS) activity of Chromobacterium violaceum. Sangyod rice seed protein hydrolysates were evaluated for their ability to inhibit the biofilm formation of bacterial cells by a crytal violet assay. Furthermore, using the dropping method, we tested the inhibitory effects of SYPs on the bacterial pigments pyocyanin in P. aeruginosa and staphyloxanthin in S. aureus. Results: Our results showed that the crude protein lysate (SYP1) did not exhibit antibacterial activity against any of the test bacteria. Intriguingly, after boiling (SYP2) and enzymatic hydrolysis (SYP3 and SYP4), the protein hydrolysates were transformed into bioactive peptides and displayed antibacterial properties against all of the test bacteria at a concentration of 500 µg as determined by agar well diffusion. SYP4 demonstrated the highest antibacterial activity as it completely inhibited all test strains, with inhibition zones ranging from 16.88 ± 0.25 to 21.25 ± 0.5 mm, and also yielded the highest MIC/MBC values against P. aeruginosa, B. cereus, and E. coli, at 256 and >256 µg/mL, respectively. We observed that at least 256 µg/mL of SYP4 is required to exhibit optimal antibacterial activity. At 16-128 µg/mL, it exhibited antibiofilm activity against S. aureus. Furthermore, at 256 µg/mL, SYP4 inhibited pyocyanin in P. aeruginosa and staphyloxanthin in S. aureus. Although SYP2 and SYP3 displayed weak antibacterial activity and their MIC values could not be obtained for all bacteria, they showed strong QS inhibition in C. violaceum at 256 µg protein. Moreover, SYP2 and SYP3, at a minimum concentration of 32 µg/mL, significantly reduced violacein production. SYP3 also showed biofilm reduction activity on S. aureus at least 16-512 µg/mL. Conclusion: Sangyod Phatthalung protein hydrolysates exerted excellent inhibitory effects against the growth of bacteria and their virulence factors, such as QS, biofilm formation, and/or pigment production. These factors include zoonotic and foodborne pathogens. Therefore, daily consumption of Sangyod Phatthalung rice might reduce the risk of bacterial pathogenesis and foodborne diseases. In conclusion, functional foods or alternate methods of treating bacterial illnesses may be developed in humans and animals.

Occurrence of multidrug resistance associated with extended-spectrum ß­lactamase and the biofilm forming ability of Escherichia coli in environmental swine husbandry.

Extended-spectrum beta-lactamase (ESBL) production and biofilm formation are mechanisms employed by Escherichia coli to resist beta-lactam antibiotics. Thus, we aimed to examine antibiotic resistance associated with ESBL production and biofilm formation in E. coli isolates from swine farms in Southern Thailand. In total, 159 E. coli isolates were obtained, with 44 isolates identified as ESBL producers, originating from feces (18.87 %) and wastewater (8.80 %) samples. All ESBL-producing strains exhibited resistance to ampicillin (100 %), followed by the cephalosporin group (97.73 %) and tetracycline (84.09 %). Multidrug resistance was observed in 17 isolates (38.63 %). Among the isolates from feces samples, the blaGES gene was the most prevalent, detected in 90 % of the samples, followed by blaCTX-M9 (86.67 %) and blaCTX-M1 (66.67 %), respectively. In the bacteria isolated from wastewater, both blaGES and blaCTX-M9 genes were the predominant resistance genes, detected in 100 % of the isolates, followed by blaCTX-M1 (64.29 %) and blaTEM (50 %), respectively. Strong biofilm formation was observed in 11 isolates (36.67 %) from feces and 4 isolates (25.57 %) from wastewater samples. Notably, nearly 100 % of ESBL-producing strains isolated from feces tested positive for both pgaA and pgaC genes, which play a role in intracellular adhesion and biofilm production. These findings contribute to the understanding and potential control of ESBL-producing E. coli, and the dissemination of antibiotic resistance and biofilm-related genes in swine farms.

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.

Development of a microencapsulated probiotic containing Pediococcus acidilactici WU222001 against avian pathogenic Escherichia coli.

Background and Aim: Probiotics are beneficial microorganisms for humans and animals. In this study, we developed a microencapsulated probiotic with antibacterial activity against avian pathogenic Escherichia coli (APEC). Materials and Methods: Alignment of the 16S rRNA sequences of the isolate WU222001 with those deposited in GenBank revealed that the isolate was Pediococcus acidilactici with 99.6% homology. This bacterium was characterized as a probiotic based on its tolerance toward in vitro gastrointestinal tract (GIT) conditions, hydrophobicity, and auto-aggregation. The antibacterial activity of the probiotic's culture supernatant against APEC was investigated using a broth microdilution assay. Pediococcus acidilactici was microencapsulated using sodium alginate and agar with diameters ranging from 47 to 61 µm. Then, physicochemical characteristics and stability of the microcapsules were determined. Results: The isolate was characterized as a probiotic based on its resistance to low pH, bile salts, and pancreatin, with relative values of 79.2%, 70.95%, and 90.64%, respectively. Furthermore, the bacterium exhibited 79.56% auto-aggregation and 55.25% hydrophobicity at 24 h. The probiotic's culture supernatant exhibited strong antibacterial activity against clinical APEC isolates with minimum inhibitory concentration and minimum bactericidal concentration of 12.5% and 25% v/v, respectively. Microencapsulation-enhanced bacterial viability in GIT compared to free cells. Moreover, 89.65% of the encapsulated cells were released into the simulated intestinal fluid within 4 h. The viable count in microcapsules was 63.19% after 3 months of storage at 4°C. Conclusion: The results indicated that the culture supernatant of P. acidilactici inhibited the growth of APEC. In addition, microencapsulation extends the viability of P. acidilactici under harsh conditions, indicating its potential application in the feed production.

Trypanosoma lewisi in blood of Rattus rattus complex residing in human settlements, Nakhon Si Thammarat, Thailand: Microscopic and molecular investigations.

Trypanosomes are blood parasites infected in various mammals, including rats. The presence of rats in human settlements can increase the chance of Trypanosoma transmission to humans. The molecular study of multispacer in Trypanosoma spp. in naturally infected rodents in Thailand is scanty. The objective of this study was to detect Trypanosoma in the blood of the captured rats in Nakhon Si Thammarat, Thailand, using microscopic and molecular techniques. This was a cross-sectional study conducted in human settlement areas. Ninety-nine blood samples were collected using cardiac puncture. A blood sample was smeared on a glass slide and examined using a compound light microscope and a scanning electron microscope. Moreover, polymerase chain reaction was applied to detect Trypanosoma evansi and T. lewisi in the blood. An additional primer set was used to confirm the species of the detected trypanosome. Approximately 18% of the rats had positive Trypanosoma infections. All Trypanosoma-positive blood samples were matched with sequences of T. lewisi. The stumpy form of trypanosome had higher nucleus related parameters than the slender form. Interestingly, the partial sequences of the alpha-tubulin gene of T. lewisi were first reported in the naturally infected RrC in this study. Based on the results obtained, T. lewisi biology, particularly the virulent components and route of transmission, pathogenesis, and in vitro experiments, are strongly recommended for further study.

Molecular detection of Loxodontofilaria spp. in Asian elephants (Elephas maximus) from elephant training camps in Thailand.

Filarial infection is an important disease in human and animal medicine. Several filarial worms are of importance, especially nematodes in the Onchocercidae. The Asian elephant (Elephas maximus) is an endangered animal and is very important from several socio-economic and ecological aspects in Thailand. Various parasites can be found in elephants; however, data related to filarial infections in elephants is limited. The objective of this study was to detect filaria in the blood of Asian elephants in Thailand, based on a polymerase chain reaction (PCR) technique. Blood samples were collected from 208 Asian elephants and detected for filaria using PCR, targeting the region of the internal transcribed spacer 2 (ITS2), the cytochrome c oxidase subunit 1 (cox1), and the RNA polymerase II large subunit (rbp1). In total, 4.33% (9 out of 208) of the sampled elephants had Loxodontofilaria spp. DNA with 100% query coverage. In addition, the obtained cox1 and rbp1 sequences matched with Loxodontofilaria sp., Onchocerca sp., and Dirofilaria sp. There were no identified risk factors (sex, age, location, and packed cell volume) related to Loxodontofilaria infection in elephants. The analyses of the phylogeny of ITS2 sequences demonstrated that the Loxodotofilaria-positive sequences were closely related to Onchocerca dewittei japonica and Onchocerca dewittei dewittei with 100% query coverage. Notably, the concatenated phylogenetic trees of ITS2 and the cox1 and rbp1 genes were closely similar to Loxodontofilaria sp. To describe in detail the genomic DNA of Loxodontofilaria spp., other genes should be additionally studied using a more discriminatory technique, such as DNA barcoding or whole genome sequencing.

Sangyod rice bran extract enhances Lacticaseibacillus paracasei growth during the exponential phase and antibacterial activity of L. paracasei supernatant against zoonotic and foodborne pathogens.

Background and Aim: Prebiotics are a group of nutrients or compounds that are degraded by the gut microbiota, including Lacticaseibacillus paracasei. The probiotic plays an important role in adhesion to the gut and is able to produce antimicrobial substances to inhibit pathogens. This study aimed to investigate the effects of Sangyod rice bran extract on the growth promotion of L. paracasei. Furthermore, antibacterial activity of the extract and L. paracasei supernatants cultured in De Man, Rogosa and Sharpe (MRS) medium plus the extract against zoonotic and foodborne pathogens was investigated. Materials and Methods: Antibacterial activity of the crude extract and the oil from Sangyod rice bran against the pathogens, including Bacillus cereus, Staphylococcus aureus, Escherichia coli, Avian pathogenic E. coli, and Pseudomonas aeruginosa was investigated using broth microdilution assay. The effects of the crude extract and the oil on the growth and adhesion of L. paracasei were further determined. The antibacterial activity of L. paracasei supernatant cultured in the medium supplemented with the extract and the oil against the pathogens was determined by agar well diffusion assay, followed by the broth microdilution assay. Finally, the chemical constituents and antioxidant activity of the crude extract and the oil from Sangyod rice bran were investigated. Results: The crude extract and the oil from Sangyod rice bran enhanced L. paracasei growth during the exponential phase. Furthermore, the crude extract at 0.25 mg/mL significantly enhanced the adhesion of L. paracasei to the surface compared with the control. Both minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values of the crude extract against B. cereus and S. aureus were 0.5 and 1.0 mg/mL, respectively. All pathogens were sensitive to the supernatant of L. paracasei with similar MIC and MBC ranging from 12.5% v/v to 50% v/v. However, the MIC and MBC values of L. paracasei supernatant grown in MRS medium plus the crude extract and oil were not significantly different compared to the supernatant obtained from MRS alone. The crude extract had free radical scavenging activities with IC50 values at 0.61 mg/mL. Conclusion: The results suggested the potential benefits of the crude extract from Sangyod rice bran for inducing the growth and the adhesion of L. paracasei and inhibiting zoonotic and foodborne pathogens.

Characterizing the antimicrobial resistance profile of Escherichia coli found in sport animals (fighting cocks, fighting bulls, and sport horses) and soils from their environment.

Background and Aim: Antimicrobial resistance (AMR) is a significant threat to global health and development. Inappropriate antimicrobial drug use in animals cause AMR, and most studies focus on livestock because of the widespread use of antimicrobial medicines. There is a lack of studies on sports animals and AMR issues. This study aimed to characterize the AMR profile of E. coli found in sports animals (fighting cocks, fighting bulls, and sport horses) and soils from their environment. Materials and Methods: Bacterial isolation and identification were conducted to identify E. coli isolates recovered from fresh feces that were obtained from fighting cocks (n = 32), fighting bulls (n = 57), sport horses (n = 33), and soils from those farms (n = 32) at Nakhon Si Thammarat. Antimicrobial resistance was determined using 15 tested antimicrobial agents - ampicillin (AM), amoxicillin-clavulanic acid, cephalexin (CN), cefalotin (CF), cefoperazone, ceftiofur, cefquinome, gentamicin, neomycin, flumequine (UB), enrofloxacin, marbofloaxacin, polymyxin B, tetracycline (TE), and sulfamethoxazole/trimethoprim (SXT). The virulence genes, AMR genes, and phylogenetic groups were also examined. Five virulence genes, iroN, ompT, hlyF, iss, and iutA, are genes determining the phylogenetic groups, chuA, cjaA, and tspE4C2, were identified. The AMR genes selected for detection were blaTEM and blaSHV for the beta-lactamase group; cml-A for phenicol; dhfrV for trimethoprim; sul1 and sul2 for sulfonamides; tetA, tetB, and tetC for TEs; and qnrA, qnrB, and qnrS for quinolones. Results: The E. coli derived from sports animals were resistant at different levels to AM, CF, CN, UB, SXT, and TE. The AMR rate was overall higher in fighting cocks than in other animals, with significantly higher resistance to AM, CF, and TE. The highest AMR was found in fighting cocks, where 62.5% of their isolates were AM resistant. In addition, multidrug resistance was highest in fighting cocks (12.5%). One extended-spectrum beta-lactamase E. coli isolate was found in the soils, but none from animal feces. The phylogenetic analysis showed that most E. coli isolates were in Group B1. The E. coli isolates from fighting cocks had more virulence and AMR genes than other sources. The AMR genes found in 20% or more of the isolates were blaTEM (71.9%), qnrB (25%), qnrS (46.9%), and tetA (56.25%), whereas in the E. coli isolates collected from soils, the only resistance genes found in 20% or more of the isolates were blaTEM (30.8%), and tetA (23.1%). Conclusion: Escherichia coli from fighting cock feces had significantly higher resistance to AM, CF, and TE than isolates from other sporting animals. Hence, fighting cocks may be a reservoir of resistant E. coli that can transfer to the environment and other animals and humans in direct contact with the birds or the birds' habitat. Programs for antimicrobial monitoring should also target sports animals and their environment.

Detection of antibodies against three zoonotic Bartonella spp. and cross-reactivity among species and Coxiella burnetii in dogs and cats from Central Thailand.

Dogs and cats are important reservoir hosts of bacterial zoonotic pathogens, especially the Proteobacteria, Bartonella spp., and Coxiella burnetii. Bartonella spp. and C. burnetii are Gram-negative intracellular bacteria causing cat-scratch disease and query fever, respectively. Despite these two pathogens being dangerous, studies of their seroprevalence and cross-reactivity are limited in Thailand. The objectives of this study were to detect the seroprevalence of three zoonotic species of Bartonella and to evaluate cross-reactivity among Bartonella spp. and with C. burnetii. In total, 570 dog and cat serum samples were detected for antibodies against Bartonella spp. and C. burnetii using an indirect immunofluorescence antibody (IFA) test. At titer ≥ 1:64, tested serum that had a fluorescent intensity score ≥ 2 was interpreted as positive. Additionally, possible factors related to the seroprevalence were analyzed consisting of sex, breed, age, residing area, and ectoparasite control. Overall, the seroprevalence levels of Bartonella spp. and C. burnetii were 13.16% and 1.23%, respectively. All antigens of Bartonella were reacted to sera (1.23-7.72%), furthermore, both phases of C. burnetii were revealed in sera (0.35-1.05%). Interestingly, there was a poor agreement of cross-reactivity among Bartonella spp. at 5.56-8.70%, while cross-reactivity between Bartonella spp. and C. burnetii also showed poor agreement (2.80%). It is suggested that dogs and cats are important reservoirs of Bartonella spp., even in animals with ectoparasite control. The Bartonella seroprevalence was high in pure-breed animals with ectoparasite control, reflecting that Bartonella spp. infections can occur in owned, well-cared-for, and asymptomatic dogs and cats.

Molecular Detection of Bartonella Species in Rodents Residing in Urban and Suburban Areas of Central Thailand.

Bartonella spp. are Gram-negative zoonotic bacteria transmitted to humans via various blood-sucking arthropods. Rodents have been identified as reservoir hosts of several zoonotic pathogens, including Bartonella spp. In Thailand, studies of Bartonella spp. in rodents from urban areas are limited; thus, a study in this area is necessary. The objectives of this study were to detect Bartonella spp. in rodents in Thailand and to compare the species' distribution across different areas. In total, 70 blood samples from rodents in urban and suburban areas were tested for Bartonella spp. using a conventional polymerase chain reaction that targeted the citrate synthase (gltA) gene. All Bartonella-positive sequences were analyzed using polymorphism in order to build a phylogenetic tree. Approximately 38% of the rodents studied contained Bartonella DNA. Both Rattus exulans (Pacific rat) and R. tanezumi (Asian house rat) contained Bartonella spp. Four species of Bartonella were detected in blood samples: B. tribocorum, B. phoceensis, B. grahamii, and B. rattimassiliensis. In addition, eight Pacific rats contained the B. kosoyi-B. tribocorum complex. Bartonella phoceensis and B. tribocorum-B. kosoyi complexes were found in a specific habitat (p < 0.05). Interestingly, only seven haplotypes were identified in the sequences analyzed, and only haplotype A was found in both rodent species. Finally, a monitoring program for zoonotic Bartonella infection, especially the B. kosoyi-B. tribocorum complex, B. phoceensis, B. grahamii, and B. rattimassiliensis should be established, especially in high-risk areas.

Detection of zoonotic Bartonella species in ticks and fleas parasitizing free-ranging cats and dogs residing in temples of Bangkok, Thailand.

Bartonellosis is a vector-borne disease caused by intraerythrocytic bacteria known as Bartonella spp. The potential vectors that transmit Bartonella spp. are fleas, ticks, sand flies, and lice. Several Bartonella spp. cause diseases in humans; however, there is few molecular evidence of Bartonella spp. in vectors in Thailand. The objectives of this study were to investigate Bartonella spp. and to evaluate the spatial distribution of Bartonella spp. prevalence in the ectoparasites parasitizing free-ranging cats and dogs in temple clusters of Bangkok, Thailand. In total, 343 ectoparasites were studied to extract their genomic DNA. Species of all specimens were identified using an identification key and conventional polymerase chain reaction (cPCR) was applied to confirm flea and tick species. Extracted DNA samples were processed using primers that targeted the gltA, rpoB, ftsZ, and ribC genes of Bartonella spp. Then, PCR-positive amplicons were sequenced and a phylogenetic tree was constructed. Recorded data were statistically analyzed using descriptive statistics, the chi-square test, Fisher's exact test, and the odds ratio. Area data were analyzed and a prevalence distribution map was plotted. The major parasitizing ticks and fleas in this study were Rhipicephalus sanguineus and Ctenocephalides felis, respectively. Overall, the prevalence of Bartonella spp. in ectoparasites was 7.00%. The gltA amplicons revealed the presence of B. henselae (4.78%) and B. clarridgeiae (4.78%) in C. felis, and B. koehlerae (1.25%) and B. phoceensis (1.25%) in R. sanguineus. Bartonella DNA was encountered in 16/39 (41.03%) districts and 28.57% of the temple clusters. Bang Khun Thian district had the highest positive proportion and Bang Bon district showed co-evidence of different Bartonella species. In addition, the intervening zones were a risk factor of Bartonella (p < 0.05), and the distribution map showed a scattered pattern of Bartonella-positive clusters. Finally, fleas showed to be important vector reservoirs for Bartonella spp., especially zoonotic species, however, experimental studies are needed to prove the Bartonella transmission in ticks.

Molecular Detection of Bartonella quintana among Long-Tailed Macaques (Macaca fascicularis) in Thailand.

Bartonella quintana is a zoonotic pathogen with a worldwide distribution. Humans and non-human primates are considered to be natural reservoir hosts for B. quintana. However, information on the molecular epidemiology of this organism is very limited in regard to long-tailed macaques (Macaca fascicularis) in Thailand. Therefore, this study aimed to investigate the occurrence and genetic diversity of Bartonella spp. among long-tailed macaques in Thailand. In total, 856 blood samples were collected from long-tailed macaques in Thailand. All specimens were screened for Bartonella spp. using a polymerase chain reaction (PCR) assay targeting the 16S rRNA, gltA and ftsZ genes. All positive samples were further analyzed based on nucleotide sequencing, phylogenetic analysis and multiple sequence alignment analysis. Only one macaque showed a positive result in the PCR assays based on the 16S rRNA, gltA and ftsZ genes. Nucleotide sequencing and phylogenetic analysis revealed that the obtained sequences were closely related to B. quintana previously detected in non-human primates. Single-nucleotide polymorphisms (SNPs) were detected in the gltA and ftsZ gene sequences. This study revealed that long-tailed macaques in Thailand carried B. quintana. Despite the low infection rate detected, long-tailed macaques may be a reservoir of B. quintana.

Molecular Detection of Bartonella spp. and Hematological Evaluation in Domestic Cats and Dogs from Bangkok, Thailand.

(1) Background: Bartonella spp. are Gram-negative, facultative, intracellular bacteria transmitted by hematophagous insects. Several species cause zoonotic diseases such as cat-scratch disease. Bartonella henselae and Bartonella clarridgeiae are the main species found in Thailand, however, there have been few studies on Bartonella spp. In addition, the hematological evaluation of Bartonella-infected animals is limited in Thailand. The aims of this study were prevalence investigation and hematological evaluation of Bartonella-infected dogs and cats residing in Bangkok, Thailand. (2) Methods: In total, 295 dogs and 513 cats were molecularly evaluated to detect Bartonella spp. using PCR with primers targeting the partial gltA, rpoB, ftsZ, ribC, and groEL genes. In total, 651 domestic animals were evaluated for hematological parameters compared between Bartonella-positive and Bartonella-negative animals. (3) Results: Overall, the prevalence of Bartonella spp. was 1.61% which was found only in free-ranging cats (2.83%). Bartonella henselae and B. clarridgeiae were confirmed from a concatenated phylogenetic tree of partial gltA and ribC genes, with 100% bootstrapping replication. For other housekeeping gene sequences, mixed infection was expected from the amplicons of rpoB, ftsZ, and groEL. Importantly, the mean corpuscular volume (MCV) was significantly increased in Bartonella-positive cats. (4) Conclusions: We suggest that B. henselae and B. clarridgeiae are important species and are still circulating in domestic animals, especially cats. The evaluation of blood parameters, especially a raised MCV, should be of concern in Bartonella infection in asymptomatic cats. Additionally, the knowledge of how to prevent Bartonella-related diseases should be promoted with people in at-risk situations.

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.