Antimicrobial resistance profiles in bacterial species isolated from fecal samples of free-ranging long-tailed macaques (Macaca fascicularis) living in Lopburi Old Town, Thailand.

BACKGROUND AND AIM: At present, increasing in long-tailed macaques (Macaca fascicularis) population in Lopburi old town caused several problems in its community, in particular with sanitation problem. The present study aimed to explore species distribution and antimicrobial resistance patterns in bacteria isolated from feces of the free-ranging long-tailed macaques (Macaca fascicularis) in Lopburi Old Town, Thailand. MATERIALS AND METHODS: Fresh fecal samples were collected from October 2018 to July 2019 from seven troops of macaques. Bacterial colonies were identified based on Gram stain and standard biochemical techniques. Sensitivity toward eight different antibiotics, including amoxicillin, amoxicillin-clavulanate, cephalexin, clindamycin, doxycycline, enrofloxacin, erythromycin, and gentamicin, was analyzed using the disk diffusion method. RESULTS: A total of 1050 fecal samples were collected. Five unique bacterial species were identified, including Escherichia coli, Enterobacter spp., Proteus spp., Salmonella Group B, and Citrobacter spp. in 100%, 25.71%, 18%, 1.71%, and 0.57% of the fecal specimens, respectively. Among 70 distinct isolates of E. coli, 63 (93%) were resistant to multiple drugs, including amoxicillin, cephalexin, clindamycin, and erythromycin; one isolate (6%) was resistant to clindamycin only. Furthermore, 17 isolates (94%) of Salmonella Group B were resistant to both clindamycin and erythromycin. Five of the six Citrobacter spp. isolates (83%) were also multidrug-resistant (to cephalexin, clindamycin, and erythromycin); the one remaining Citrobacter spp. isolate (6%) was resistant to both clindamycin and erythromycin. However, a high percentage of E. coli, Salmonella Group B and Citrobacter spp. remained susceptible to amoxicillin-clavulanate, enrofloxacin, and doxycycline. CONCLUSION: Our findings provide the basic information for the selection of empirical therapy and for the evaluation of the scale of antibiotic resistance associated with macaques living in Lopburi Old Town.

Seroprevalence of Dengue, Zika, and Chikungunya Viruses in Wild Monkeys in Thailand.

Zoonotic pathogens such as arboviruses have comprised a significant proportion of emerging infectious diseases in humans. The role of wildlife species as reservoirs for arboviruses is poorly understood, especially in endemic areas such as Southeast Asia. This study aims to determine the exposure history of different macaque species from national parks in Thailand to mosquito-borne flaviviruses and alphavirus by testing the serum samples collected from 25 northern pigtailed macaques, 33 stump-tailed macaques, and 4 long-tailed macaques for the presence of antibodies against dengue, Zika, and chikungunya viruses by plaque reduction neutralization assay. Specific neutralizing antibodies against Dengue virus (DENV1-4) and Zika virus (ZIKV) were mainly found in stump-tailed macaques, whereas neutralizing antibody titers were not detected in long-tailed macaques and pigtailed macaques as determined by 90% plaque reduction neutralization assay (PRNT90). One long-tailed macaque captured from the south of Thailand exhibited antibody titers against chikungunya virus (CHIKV), suggesting enzootic of this virus to nonhuman primates (NHPs) in Thailand. Encroachment of human settlements into the forest has increased the interface that exposes humans to zoonotic pathogens such as arboviruses found in monkeys. Nonhuman primates living in different regions of Thailand showed different patterns of arboviral infections. The presence of neutralizing antibodies among wild monkeys in Thailand strongly suggests the existence of sylvatic cycles for DENV, ZIKV, and CHIKV in Thailand. The transmission of dengue, Zika, and chikungunya viruses among wild macaques may have important public health implications.

Comparison of Rabbit, Kitten and Mammal Milk Replacer Efficiencies in Early Weaning Rabbits.

Early weaned rabbits should be fed using a milk replacer in order to survive. Therefore, a rabbit milk replacer (RMR) was developed and compared with a kitten milk replacer (KMR®: KMR) and a mammal milk replacer (Zoologic® Milk matrix 30/52: MMR). Thirty-six native crossbred rabbits aged 18 days were divided into three experimental groups (six replicates/group, two rabbits/replicate), fed RMR, KMR or MMR daily until they were 36 days old and euthanized at 38 days, while a complete pelleted diet and water were provided ad libitum. No statistically significant differences were observed in growth performance parameters, water intake, faecal weight, nutrient digestibility, internal organ weight, caecal pH, caecal cellulose activity, number of faecal pellets and amount of crude protein intake (p > 0.05). Caecal amylase activity in the KMR group and caecal protease activity in the RMR group were higher than in the MMR group (p < 0.05). The villus height and crypt depth of the MMR group were greater than in the RMR and KMR group (p < 0.05). In conclusion, it is possible to feed RMR to early weaning rabbits without serious adverse effects. However, probiotics and/or prebiotics should be supplemented in milk replacers and their benefits studied.

First detection and characterization of Psittaciform bornaviruses in naturally infected and diseased birds in Thailand.

In Thailand a proventricular dilation disease (PDD)-like syndrome commonly occurs in captive psittacine birds. The etiology, however, has been unknown to date and studies to detect parrot bornaviruses have never been performed in Southeastern Asia. Therefore, 111 psittacines (22 different species) including birds with suspected PDD based on clinical examination results (n = 65), cage mates of PDD suspected parrots without any clinical signs (n = 39) and dead birds with previous clinic suspicious for PDD (n = 7) were tested for bornaviruses using various reverse transcription polymerase chain reaction (RT-PCR) and realtime RT-PCR protocols, an enzyme-linked immunosorbent assay (ELISA), immunohistochemistry, and genome sequencing. Bornaviral infections, indicated by the presence of RNA or antibody positive reactions were detected in 60 birds (54.1%) belonging to 15 psittaciform species and originating from 41 owners. Occurrence of Psittaciform 1 orthobornavirus was confirmed by sequencing of PCR products in 24 of these birds. Parrot bornavirus (PaBV)-5, belonging to the species Psittaciform 2 orthobornavirus and found only in single birds in the United States of America, Japan and Hungary until now, was identified in a macaw. Full genome sequencing revealed features shared with other strains of this virus. PaBV-4 was the prevalent virus type and the viruses grouped in two of the five genetic PaBV-4 subclusters known so far while PaBV-2 was found in a single patient. Forty-five psittacines of the group of PDD-suspected birds (69.2%), 4 dead birds and 11 clinically healthy cage mates were positive in at least one test the latter suggesting inefficient horizontal transmission in natural infections. Lymphoplasmacytic infiltrations (non-purulent inflammation, ganglioneuritis) and bornavirus antigen were detected in diverse tissues confirming PDD as the disease involved. These results may have a major impact on conservation projects including the five near-threatened parrot species living in the wild in Thailand.

Molecular Identification of Cryptosporidium Species from Pet Snakes in Thailand.

Cryptosporidium is an important pathogen causing gastrointestinal disease in snakes and is distributed worldwide. The main objectives of this study were to detect and identify Cryptosporidium species in captive snakes from exotic pet shops and snake farms in Thailand. In total, 165 fecal samples were examined from 8 snake species, boa constrictor (Boa constrictor constrictor), corn snake (Elaphe guttata), ball python (Python regius), milk snake (Lampropeltis triangulum), king snake (Lampropeltis getula), rock python (Python sebae), rainbow boa (Epicrates cenchria), and carpet python (Morelia spilota). Cryptosporidium oocysts were examined using the dimethyl sulfoxide (DMSO)-modified acid-fast staining and a molecular method based on nested-PCR, PCR-RFLP analysis, and sequencing amplification of the SSU rRNA gene. DMSO-modified acid-fast staining revealed the presence of Cryptosporidium oocysts in 12 out of 165 (7.3%) samples, whereas PCR produced positive results in 40 (24.2%) samples. Molecular characterization indicated the presence of Cryptosporidium parvum (mouse genotype) as the most common species in 24 samples (60%) from 5 species of snake followed by Cryptosporidium serpentis in 9 samples (22.5%) from 2 species of snake and Cryptosporidium muris in 3 samples (7.5%) from P. regius.

Anterior ocular abnormalities of captive Asian elephants (Elephas maximus indicus) in Thailand.

OBJECTIVE: To survey and classify anterior ocular abnormalities in 1478 captive Asian elephants (Elephas maximus indicus) in six regions of Thailand. METHODS: Anterior ocular examination was performed in both eyes (n = 2956) of 1478 elephants selected from the annual health check program involving 2958 animals within six regions of Thailand from January to November 2013. Lesions were described and compared between age and gender. RESULTS: A total of 17.83% (527/2956) of examined eyes from 24.97% (369/1478) of examined elephants had anterior ocular abnormalities. The most common lesions in these examined eyes were frothy ocular discharge (5.85%), corneal edema (5.31%), and conjunctivitis (5.18%). In addition, epiphora, phthisis bulbi, other corneal abnormalities, anterior uveitis, and lens abnormalities were noted. Almost all lesions increased in frequency with age (P < 0.01). CONCLUSIONS: Regular ophthalmic examination in elephants should be included in their annual health check program. Early detection and treatment of any ocular abnormality may avoid the development of subsequent irreversible ocular pathology.

Quantitative and qualitative morphologic, cytochemical, and ultrastructural characteristics of blood cells in captive Asian water monitors.

BACKGROUND: The Asian water monitor (Varanus salvator) is the most common monitor lizard in Thailand. Reported data regarding hematology and morphology of blood cells for this species are scarce. OBJECTIVES: The objective of this study was to assess routine hematologic variables and characterize the morphology, cytochemical staining, and ultrastructural features of blood cells in the Asian water monitor. METHODS: Blood samples from 55 monitors (22 males and 33 females) were obtained for a CBC. Cytochemical staining (Sudan black B [SBB], peroxidase [PO], α-naphthyl acetate esterase [ANAE], and beta-glucuronidase [BG]), and scanning and transmission electron microscopy were performed using standard methods. RESULTS: Determined mean (range) hematologic results of all monitors included PCV 0.32 L/L (0.20-0.44 L/L), HGB 106 g/L (62-157 g/L), WBC 15.9 × 10(9) /L (4.0-34.0 × 10(9) /L), heterophil 6.3 × 10(9) /L (1.5-17.1 × 10(9) /L, azurophil 2.6 × 10(9) /L (0.7-9.5 × 10(9) /L), basophil 0.1 (0.1-0.5 × 10(9) /L), lymphocyte 6.8 × 10(9) /L (0.5-13.1 × 10(9) /L), and monocyte 0.2 × 10(9) /L (0.04-1 × 10(9) /L) counts. Heterophils and basophils stained strongly positive with SBB, ANAE, and BG. Heterophils contained 2 types of granules, round SBB-positive and PO-negative granules, and electron-dense, large rod-shaped granules. Gamonts of Hepatozoon sp. were found in <1% RBC of 43 monitors. There was no significant difference between hematologic variables in Hepatozoon-positive and -negative monitors. CONCLUSION: Heterophils in Asian water monitors may also function as eosinophils based on cytochemical and ultrastructural features. The quantitative results may be used as base for further studies in healthy and diseased Asian water monitors.